CZ20004784A3 - Beta-lactam compounds and their use as tryptase inhibitors - Google Patents

Abstract

Compounds of formulas (I), (II), (III), (IV), (V) and (VI). These compounds inhibit tryptase as well as other enzyme systems, or they are selective tryptase inhibitors, and are useful as anti-inflammatory agents, particularly in the treatment of chronic asthma.

Description

Beta-lactam compounds and their use as tryptase inhibitors

Technical field

The invention relates to beta-lactam compounds and their use as tryptase inhibitors.

State of the art

In U.S. Patents Nos. 5,037,819, 5,110,812, 5,175,283, _5,250,677 and 5,326,863, the author Han discloses 3-guanidinoalkyl-2-azetidinones of the general formula

N-W Η H

U- N— C- N— CH ₂ — (CH ₂ ) n— CH.2 ¹ i

H ^H y

Where

U and W independently represent hydrogen or an amino protecting group;

n represents an integer from 1 to 3;

X represents a group selected from the group consisting of hydrogen, trialkylsilyl, arylsulfonyl, amino-substituted arylsulfonyl, alkylsulfonyl, arylaminocarbonyl, alkylcarbonyl, and arylcarbonyl;

_Y represents a group from the group consisting of hydrogen, arylalkenyl, arylalkyl, formyl, carboxy, alkoxycarbonyl, acyloxy, arylthio, arylsulfinyl, arylsulfonyl, alkylthio, alkylsulfinyl, alkylsulfonyl, arylaminocarbonyl, or

R represents hydrogen, alkyl or arylalkyl group;

m is an integer from 1 to 3; and

R' represents hydrogen or -CO2R'', where R'' represents hydrogen, an alkyl or an arylalkyl group.

In the writings of the author Han it is further stated that the above-mentioned compounds in which

U and W represent hydrogen;

X represents a group including arylsulfonyl, amino-substituted arylsulfonyl, alkylsulfonyl, arylaminocarbonyl, alkylcarbonyl, and arylcarbonyl; and

Y represents a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, acyloxy, arylsulfonyl, alkylthio, alkylsulfonyl, arylaminocarbonyl, o

II

C—OR

O —c— nh-ch ₂ — or

( _CH2 ) _m

R represents hydrogen, alkyl or arylalkyl group;

R' represents hydrogen or -CO2R'';

R'' is hydrogen, an alkyl or arylalkyl group, and their pharmaceutically acceptable salts, are inhibitors of serine proteases, particularly thrombin and trypsin, and can be used to control blood coagulation or to treat pancreatitis.

In the above-mentioned references, Han states that aryl means a phenyl or naphthyl group which is unsubstituted or substituted with one or more groups selected from the group consisting of amino, nitro and alkyl, and that amino may mean an unsubstituted group or a group substituted with one or two alkyl groups.

The essence of the invention

The invention relates to novel beta-lactam compounds of general formulae I, II, III, IV, and V, shown below, and the use of these compounds as in vivo inhibitors of various enzyme systems including tryptase, thrombin, trypsin, factor Xa, factor VIIa, and urokinase-type plasminogen activator. The invention also includes the use of compounds of general formula VI shown below as inhibitors of tryptase, factor Xa, factor VIIa, and urokinase-type plasminogen activator.

Compounds of the invention include compounds of general formula (I):

GO

H ₂ N— C—Αχ·

Λ -1 Ί

O

-Ri

Xi • · · · · * ·

R 1 represents a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, A ₂ -aryl,

O

II —c—r ₇ ^{0 11} I — CN- — ¹ (CHz),

Re

O

II

-C-0-R7

O ^<CH ₂ ) _v -x ί Y — c— N /

In trouble

Rs.

C—CH ₂ —O-R10

-SO2-R7,

.. T..

—CNA ₂ -n-CR ₄ , or —CNA ₂ -N—C—OR ₄ , —C—alkyl—SO ₂ —R ₇ —C-aryl—SO ₂ -R ₇ or R 1 represents an alkyl group with the proviso that R ₂ represents an alkyl group and R 3 represents hydrogen.

R2 and R3 both represent hydrogen, or _R2 represents an alkyl group with the proviso that in this case R3 represents hydrogen, or R3 represents an alkyl group with the proviso that in this case _R2 represents hydrogen.

Xi means

O

II c—R ₇ o I ¹¹ — CN- — ¹ (CHz),

O

II cRe ^(CH ₂ ) _v ·^

N ¥

V(CH ₂ ) _w >

r ₈

O (CH ₂ ) _o .Βχ, (CH ₂ )n ^ ^B 2

B ₃ — C—alkyl—so ₂ —r ₇ ° ⁰

II _n II c aryl so ₂ —r ₇ c ch ₂ - 0 Rio o

—SO ₂ -R ₇ ' c—NH SO ₂ —R ₇

Rg Rg O •i ¹ i H li νθ ? ⁹ ií cna ₂ -ncr ₄ ^{1 11 5} í?

or CN A ₂ ~N— C ^— OR ₄

-<CH ₂ )r-ΤΤΛ (CH ₂ ) _s ·

A1 means -(CH2)p-,

-N (CH ₂ ) _q — -nh— (CH ₂ ) _r R 20 /(CH ₂ ) _t (CH ₂ ) _s - ,

-\ů \/\ ₍ CH ₂ ) _u - ' -HN-CH 2 -, -HN-(CH ₂ ) 2 , or -HN-(CH ₂ ) 6 -.

R ₄ and R 5 independently represent a group selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A ₂ -cycloalkyl, A 2 -substituted cycloalkyl, aryl, substituted aryl, A ₂ -aryl,

A ₂ -substituted aryl, aryl-A3-ary1, A-ary1-A ₃ ~aryl, heteroaryl, A2-heteroary1, heterocycloalkyl,

A2~heterocycloalkyl1, ary1-A3-cycloalkyl,

A2-aryl-A ₃ -cycloalkyl, ary1-A3~heteroary1, aryl-A ₃ -heterocycloalkyl, A ₂ -aryl-A ₃ -heteroary1, A ₂ -aryl-A ₃ -heterocycloalkyl1, cycloalkyl-A ₃ -aryl, A2-cycloalkyl-A ₃ -aryl, cycloalkyl-A3-cycloalkyl, A ₂ -cycloalkyl-A ₃ -cycloalkyl1, cycloalkyl-A3-heteroaryl, A2-cycloalkyl- _A3- heteroaryl, cycloalkyl- _A3- heteroaryl1, cycloalkyl- _A3- heterocycloalkyl and « ·

A2-cycloalkyl1-A3-heterocycloalkyl.

R 0 represents a group from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2 ^- substituted cycloalkyl, aryl, substituted aryl, A2-aryl, A2-substituted aryl, aryl-A3-aryl, A2 ^- aryl-A3-aryl, heteroaryl, A2-heteroaryl, heterocycloalkyl, A2-heterocycloalkyl, aryl-A3-cycloalkyl, A2-aryl-A3-cycloalkyl, aryl-A3-heteroaryl,

A2-aryl-A3-heteroaryl, aryl-A3~heterocycloalkyl1,

A2 ^- Aryl-A3 ^- Heterocycloalkyl, Carboxy, Alkoxycarbonyl, Aryloxycarbonyl, Alkoxycarbonylamino, Aryloxycarbonylamino, Arylcarbonylamino, -N(alkyl)(Alkoxycarbonyl), -N-(Alkyl)(Aryloxycarbonyl), Alkylcarbonylamino, -N(Alkyl)(Alkylcarbonyl) and -N-(Alkyl)(Arylcarbonyl).

m means an integer from 1 to 5

Y stands for 0, S, NR* , N-SO2-R7, N—C—A3—R ₇

N

NC—R ₇

II

N-C-

II or N—CII

II _K-CC—r ₇

R7 represents a group from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, aryl, substituted aryl, A2-aryl, A2-substituted aryl, heteroaryl, A2-heteroaryl, heterocycloalkyl,

A2^_heterocycloalkyl, ary1-A3~ary1, A2-aryl-A3~ary1, ary1-A3~cycloalky1, A2-aryl-A3~cycloalkyl, ary1-A3~heteroary1, A2~aryl-As-heteroary1, aryl-A3-heterocycloalkyl, A2-aryl-A3-heterocycloalkyl, ary1-A3~substituted aryl, A2-aryl-A3~substituted aryl, ary1-A3~substituted cycloalkyl, A2-aryl-A3-substituted cycloalkyl, cycloalkyl-A3-cycloalkyl, A2-cycloalkyl-A3~cycloalkyl, cycloalkyl-A3-aryl, A2~cycloalkyl-A3-aryl, cycloalkyl-A3^_heteroaryl, A2~cycloalkyl1-A3^_heteroaryl, cycloalkyl-A3~heteroaryl, cycloalkyl-A3-heterocycloalkyl, A2-cycloalkyl-A3-heterocycloalkyl, cycloalkyl-A3~substituted cycloalkyl, A2-cycloalkyl-A3-substituted cycloalkyl, cycloalkyl-A3~substituted aryl, A2-cycloalkyl-A3~ -substituted aryl, substituted cycloalkyl-A3^_cycloalkyl, A2-substituted cycloalkyl-A3-cycloalkyl, substituted cycloalkyl-A3-substituted cycloalkyl, A2-substituted cycloalkyl-A3-substituted cycloalkyl, substituted • φ 0 • · · · · cycloalkyl-A3-aryl, A2-substituted cycloalkyl-A3-aryl, substituted cycloalkyl-A3-heteroaryl, A2-substituted cycloalkyl-A3-heteroaryl, substituted cycloalkyl-A3~ -heterocycloalkyl, A2-substituted cycloalkyl-A3 -heterocycloalkyl, substituted cycloalkyl-A3-substituted aryl, A2-substituted cycloalkyl-A3-substituted aryl, heteroaryl-A3-heteroaryl, A2-heteroaryl-A3-heteroaryl, heteroaryl-A3-cycloalkyl, A2-heteroaryl-A3-cycloalkyl, heteroaryl-A3-substituted cycloalkyl, A2-heteroaryl-A3-substituted cycloalkyl, heteroaryl-A3-aryl,

A2-heteroaryl-A3-ary1, heteroaryl-A3~heterocycloalkyl, A2-heteroaryl-A3-heterocycloalkyl, heteroaryl-A3~substituted aryl, A2~heteroaryl-A3-substituted aryl, heterocycloalkyl-A3-heterocycloalkyl, A2~heterocycloalkyl-A3-heterocycloalkyl, heterocycloalkyl-A3~ -cycloalkyl, heterocycloalkyl-A3~substituted cycloalkyl, A2-heterocycloalkyl-A3-substituted cycloalkyl, heterocycloalkyl-A3~aryl, A2-heterocycloalkyl-A3-aryl, heterocycloalkyl-A3-substituted aryl, A2-heterocycloalkyl-A3~substituted aryl, heterocycloalkyl-A3-heteroary1, A2~heterocycloalkyl-A3~heteroaryl, substituted aryl-A3~

-substituted aryl, A2~substituted aryl-A3 ^- substituted aryl, substituted aryl1-A3~cycloalkyl, A2~substituted aryl-A3~cycloalkyl, substituted aryl-A3-substituted cycloalkyl, A2~substituted aryl-A3~substituted cycloalkyl, substituted aryl1-A3~aryl, A2~substituted aryl-A3~aryl, substituted aryl-A3-heteroaryl, A2-substituted aryl-A3~ -heteroaryl, substituted aryl-A3-heterocycloalkyl,

A2-substituted aryl-A3-heterocycloalkyl,

Z or ^A2 nao means 1 or 2 with the proviso that the sum of nao is 2 or 3;

v and w mean 1, 2, or 3 with the proviso that the sum of v and w is 3, 4, or 5.

Re represents a group from the group consisting of hydrogen, halogen, amino, -NH(lower alkyl), -N(lower alkyl)2, nitro, alkyl, substituted alkyl, alkoxy, hydroxy, aryl, substituted aryl, A2-aryl, A2-substituted aryl, aryl-A3-aryl, A2 ^- aryl-A3-aryl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, heteroaryl, A2-heteroaryl, heterocycloalkyl, A2-heterocycloalkyl, aryl-A3-cycloalkyl, A2-aryl-A3-heteroaryl, A2 ^- aryl-A3-heteroaryl, aryl-A3-heterocycloalkyl, and A2-aryl-A3-heterocycloalkyl.

B1, B2 and B3 each represent CH, or two of B1, B2 and B3 represent CH and the third represents N, or one of B1, B2 and B3 represents CH and the other two represent N.

R9 represents hydrogen or a lower alkyl group.

R10 represents a group from the group consisting of alkyl, substituted alkyl, alkyl-O-alkyl, alkyl-O-alkyl-O-alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl,

A2-substituted cycloalkyl, aryl, substituted aryl,

A2^-aryl, A2~substituted aryl, aryl-A3~ary1, A2-aryl-A3 -aryl, heteroaryl, A2-heteroaryl1, heterocycloalkyl, A2~heterocycloalkyl, aryl-A3-cycloalkyl,

A2-aryl-A3-cycloalkyl, aryl-A3-heteroaryl, A2-aryl-A3-heteroaryl, aryl-A3-heterocycloalkyl and

A2-aryl-A3-heterocycloalkyl.

R20 represents a group from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, A2-aryl, and A2-substituted aryl.

R21 and R22 independently represent a group selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, A2-aryl, and A2-substituted aryl.

means whole number 2 up to 6; means whole number 1 up to 6; means 0, 1 or 2; means 1 or 2; means 1, 2, 3, or 4; means 1, 2 or 3.

A2 represents an alkylene or substituted alkylene bridge of 1 to 10 carbon atoms, an alkenyl or substituted alkenyl bridge of 2 to 10 carbon atoms containing one or more double bonds, or an alkynyl or substituted alkynyl bridge of 2 to 10 carbon atoms containing one or more triple bonds.

A3 represents a bond, an alkylene or substituted alkylene bridge of 1 to 10 carbon atoms, an alkenyl or substituted alkenyl bridge of 2 to 10 carbon atoms containing one or more double bonds, an alkynyl or substituted alkynyl bridge of 2 to 10 carbon atoms containing one or more triple bonds,

-(CH2)dO-( _CH2 )e-, -(CH2)dS-( _CH2 )e-, ( _CH2 )d—Y—( _CH2 )e R21 — ( _CH2 )

(CH ₂ ) _e —

-(CH ₂ ) d—N—C— N— (CH ₂ ) e—

R ₂ 1 R22 (CH ₂ )—C— ^{( ch} 2 ) e— R21 íí (CH ₂ )—θ— ^c - ^(CIÍ2 ) ^e

F-21

d and e have a value independently selected from the group consisting of zero and an integer from 1 to 6.

Compounds of the invention include compounds of general formula (II):

NH

H ₂ N“C—NH—(CH ₂ )5

- NO ¹ —] in

Rl

Where

R 1 represents an alkyl group, and

R 1 , X 1 and q have the meanings given above.

Compounds of the invention include compounds of general formula (III):

NH

H ₂ N— C —NH— (CH ₂ )

Rl2 H

q - 4

Ri

Where

R12 represents an alkyl group, and

R1, X1 and q have the meanings given above.

Compounds of the invention include compounds of general formula (IV):

where

NH

II

q -- SI

-Ri

_X2

O

Rl and q have the meaning given above, (CH ₂ ) _m

X2 means

•About

— C—N <_(CH ₂ ) _w

-c—CH 2 -O-R 10 ' -C—NH SO 2 -R 7 ' 3 i? and? í' W — C-aryl—SO ₂ -R ₇ ^{C N_A} 2“ ^{N c R} 4

C-alkyl-SO ₂ -R ₇

O p Rg O

II 1 ⁹ I II

C- N- A ₂ - N- c— 0R ₄

O

-CR ₇ with reservation,

O

II

-C-R7 does not mean a group from the group consisting of alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, and substituted naphthylaminocarbonyl, or means -SO2R7 with the proviso that -SO2R7 does not mean a group from the group consisting of alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl, and substituted naphthylsulfonyl.

Ra, Rs, Y, R6, m, η, o, B1, B2, B3, Re, R9 and R10 have the meanings given above.

Compounds of the invention include compounds of general formula (V):

where

Η ₂ Ν- C—ΝΗ-(CH ₂ ) Γ • ^R 13 χ ₃ f represents an integer from 3 to 5, .(CH ₂ ) ν'Rl 3 represents

O

II — c-

-c— CH ₂ -O-Rio —C—alkyl—SO ₂ —R ₇

R ₉ ^R 9 — C—aryl—SO ₂ —R ₇ CNA ₂ NC R4

O

-CR ₇ θ Rq θ

II ? ⁹ 1 II —CN-A2-N- C-OR4

O

-cr ₇ does not mean a group with the proviso that, from the group consisting of phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, carboxymethylaminocarbonyl, and alkoxycarbonylmethylaminocarbonyl, or means -SO2R7 with the proviso that -SO2R7 does not mean a group from the group consisting of alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl, and substituted naphthylsulfonyl, or means ⁰ I ¹¹ 1 — C N-|— ^{1 R} 6 (CH ₂ ), • · ♦ · ·«· ·· • · * » · · · «··« · ··* « · • · · · ·««»··« · • « · · · · *« « « « · · with the proviso that when m is 1, 2, or 3, then Re does not mean a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, and aryloxycarbonyl.

X3 represents a group from the group consisting of phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl and substituted naphthylsulfonyl.

R4, R5, Y, m, n, o, B1, B2, B3, v, w, and R6 have the meanings given above.

The invention also relates to the use of beta-lactam compounds of general formula IV shown below as inhibitors of tryptase, factor Xa, factor VIIa and urokinase plasminogen activator.

(VI) ,R1 ₄

H ₂ N—C—NH-(CH ₂ ) 7 x ₃ where

R14 represents a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, » ·0 * » 0 00 »00 000 000 »000 0 000 0 1 0 0 «00 4 · » 0 0 0 0 0

000000 0· 0 40 0 alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl, substituted naphthylsulfonyl, phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, A2~aryl, or □(CH2) _m

Re where m is 1, 2, or and Re is a group selected from the group consisting of hydrogen, carboxy, alkoxycarbonyl, and aryloxycarbonyl.

X3 and f have the meanings given above.

Detailed description of the invention

The term alkyl refers to straight or branched chain radicals having up to ten carbon atoms. The term lower alkyl refers to straight or branched chain radicals containing up to four carbon atoms and denotes a preferred subset of the term alkyl.

The term substituted alkyl refers to the above-mentioned straight or branched chain radicals containing 1 to 10 carbon atoms in which one or more, preferably one, two or three hydrogen atoms are replaced by a group including hydroxy, amino, cyano, halogen, trifluoromethyl, nitro, -NH(lower alkyl), -N-(lower alkyl)2, alkoxy, alkylthio, carboxy, alkoxycarbonyl, aminocarbonyl and alkoxycarbonylamino.

The term alkoxy means the alkyl groups listed above attached to an oxygen atom. The term alkylthio means the alkyl ·· 9« ·» 9 ·« • 9 I * -- · * • e»« · ? » » < · « í » · · ···#·«· 9 * · · · » » ·· *««·)» 99 9 9 <

groups as described above attached to a sulfur atom. The terms lower alkoxy and lower alkylthio mean lower alkyl groups as described above attached to an oxygen atom or a sulfur atom.

The term cycloalkyl means fully or partially saturated rings of 3 to 7 carbon atoms.

The term substituted cycloalkyl refers to the above-mentioned rings of 3 to 7 carbon atoms containing one or more substituents selected from the group including lower alkyl, lower alkoxy, lower alkylthio, halogen, hydroxy, trifluoromethyl, nitro, cyano, amino, -NH(lower alkyl), -N(lower alkyl)2, and carboxy, as well as groups where said rings are fused to a phenyl ring such as tetrahydronaphthyl.

The term aryl means a group selected from the group consisting of phenyl, 1-naphthyl and 2-naphthyl.

The term substituted aryl means phenyl,

1-naphthyl and 2-naphthyl groups containing a substituent selected from the group consisting of alkyl of 1 to 10 carbon atoms, lower alkoxy, lower alkylthio, halogen, hydroxy, trifluoromethyl, nitro, amino, -NH(lower alkyl), -N(lower alkyl)2, and carboxy, and di- and tri-substituted phenyl, 1-naphthyl or 2-naphthyl groups, where the substituents are groups from the group consisting of methyl, methoxy, methylthio, halogen and amino.

The term heteroaryl refers to unsaturated or partially saturated rings of 4 to 7 carbon atoms containing one or two O and S atoms and/or one to four N atoms, one to three N atoms if the ring consists of 4 atoms, with the proviso that the total number of heteroatoms in the ring is 4 or less, and 3 or less in the case of a ring consisting of 4 atoms. The heteroaryl ring may be attached through a carbon or nitrogen atom, which allows this. Preferred heteroaryl groups include 2-, 3-, or 4-pyridyl, 4-imidazoles, 4-thiazolyl, 2- and 3-thienyl, 2- and 3-furyl, and 2-(1,4,5,6-tetrahydropyrimidines). The term heteroaryl also includes bicyclic rings in which a 4 to 7 membered ring containing O, S, and N atoms as described above is fused to a benzene, cycloalkyl, heteroaryl, or heterocycloalkyl ring. Preferred bicyclic ring groups include 2- and 3-indolyl, and 4- and 5-quinoline. Said monocyclic or bicyclic heteroaryl ring may additionally be substituted on one or more available carbon atoms with a group selected from the group consisting of lower alkyl, halogen, carboxy, hydroxy, A2-lower alkyl, A2-guanido, benzyl, and cyclohexylmethyl. Furthermore, if the mono- or bicyclic ring contains an accessible N- atom, said N atom may be substituted with an N-protecting group such as benzyloxycarbonyl, tert-butoxycarbonyl, benzyl, or benzhydryl.

The term heterocycloalkyl refers to fully saturated rings of 4 to 7 atoms containing one or two O and S atoms and/or one to four N atoms, and one to three N atoms in the case where the ring contains 4 atoms, with the proviso that the total number of heteroatoms in the ring is 4 or less, and 3 or less in the case where the ring contains 4 atoms. Said heterocycloalkyl may be attached through a carbon or nitrogen atom which allows this. Preferred heterocycloalkyl groups include pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, morpholinyl, tetrahydro-1,2-thiazinyl, piperazinyl, piperidinyl, homopiperizinyl, and azetidinyl. The term heterocycloalkyl also includes bicyclic rings wherein a 4 to 7 membered saturated ring containing O, S and N atoms as described above is fused to a cycloalkyl, benzene, heteroaryl or heterocycloalkyl ring. Said mono- or bicyclic heterocycloalkyl ring may be further substituted on one or more accessible carbon atoms with a group selected from the group consisting of lower alkyl, halogen, carboxy, hydroxy, A2-lower alkoxy, A2-guanido, benzyl and cyclohexylmethyl. If the mono- or bicyclic heterocycloalkyl ring contains an accessible N atom, said N atom may be further substituted with an N-protecting group such as benzyloxycarbonyl, tert-butoxycarbonyl, benzyl or benzhydryl.

The term halogen means chlorine, bromine, fluorine and iodine.

The term alkylene and substituted alkylene means a bridge formed by 1 to 10 carbon atoms such as -CH2-, —(CH2)2— ,

-(CH2)9- etc. One or more hydrogen atoms in the alkylene bridge, preferably one, may be replaced by a group from the group consisting of alkyl, substituted alkyl, carboxy, alkoxycarbonyl, amino, -NH(lower alkyl), -N(lower alkyl)2, hydroxy, aminocarbonyl, alkoxycarbonylamino, halogen, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, or heterocycloalkyl, such as in the groups

'Γ ch ₃ h-ch ₂ - -ch-CH ₂ och ₃

C2H5 and the like

The term alkenyl and substituted alkenyl means a bridge formed by 2 to 10 carbon atoms containing one or more double bonds, preferably containing 2 to 6 carbon atoms with one double bond, such as -CH=CH-, -CH=CH-CH2~, -(CH ₂ )-CH=CH, etc. One or more hydrogen atoms in the alkenyl bridge, preferably one, may be replaced by a group from the group consisting of alkyl, substituted alkyl, carboxy, alkoxycarbonyl, amino, -NH(lower alkyl), -N(lower alkyl)2, hydroxy, aminocarbonyl, alkoxycarbonylamino, halogen, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, or heterocycloalkyl, such as in the groups

CH ₃ and the like.

-CH= CH— j!HC ₂ H ₅

The term alkynyl and substituted alkynyl means a bridge formed by 2 to 10 carbon atoms containing one or more triple bonds, preferably containing 2 to 6 carbon atoms with one triple bond, such as -CH2CH-, -CH2-CHC-,

-C=C-(CH2)-, etc. One or more hydrogen atoms in the alkynyl bridge may be replaced by a group from the group consisting of alkyl, substituted alkyl, carboxy, alkoxycarbonyl, amino, -NH(lower alkyl), -N(lower alkyl)2, hydroxy, aminocarbonyl, alkoxycarbonylamino, halogen, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, or • · « «

heterocycloalkyl, such as in the groups

and so on.

Compounds of general formulas IV, V, and VI can be prepared by the methods set forth below.

Azetidinone substituted with a carboxy group of the formula (VII) _>co ₂ h )-Np ₃ o

where P3 represents a silyl protecting group such as tert-butyldimethylsilyl, is treated with an alkyl dihalide of formula (VIII)

Cl-(CH ₂ )qI in the presence of a base to form a carboxy-substituted azetidinone, formula

C1-(CH ₂ ) _/CO ₂ H )—Np ₃ o (IX) • ·

The carboxy-substituted azetidinone of formula (IX) is then treated with an azide, such as sodium azide, and then with a salt containing a fluoride ion, such as tetrabutylammonium fluoride, to remove the silyl group to give the compound of formula (X)

N ₃ - (CH ₂ ) q,co ₂ h

-NH

0'

Hydrogenation of the compound of formula X by treatment with hydrogen in the presence of a catalyst, palladium on carbon, gives the alkylamine compound of formula (XI)

H ₂ N-(CH ₂ ) _ _Z CO ₂ H ty-NH

O

Said alkylamine compound of formula XI, preferably in the form of an acid salt, is reacted with a guanylating agent with two protecting groups, of formula (XII)

N-Px

P1—N—C—L H where P1 represents an N-protecting group such as tert-butoxycarbonyl or benzyloxycarbonyl, and L is a leaving group such as methylthio or pyrazolyl, to give an azetidinone compound of formula (XIII)

N-Ri

II

HN—CN—(CH ₂ ) _q I | ^p ii , co ₂ H

NH

By coupling the intermediate compound of formula XIII with an amine selected from the group consisting of ch ₂ )

HN^ r ₅

HN~ |— ¹ Re ^(CH ₂ ) _v hn ^y (CH ₂ )^ (CH ₂ )o / V

HN\

R9 Rg O

I I II

HN—A ₂ —N—c—R ₄ (CH ₂ )n ^ ^B 2 b ₃ r ₈ or

R9 Rg O

I 1 11

HN—A ₂ —N—C-OR ₄ gives a compound of the formula

N-Pill

HN—C—N—(CH ₂ ) _q Rl5

-NH where R15 means (XIV) • · · ·

θ R9 r ₉ o

II I i ii — c- N—A ₂ —N— c—R ₄ or

OR ₉ R ₉ O

II II II cn—a ₂ —n—c—O-R4 .

By reacting the intermediate of formula XIV with an acid chloride selected from the group consisting of (CH ₂ ), ° o „ Cl—CN-|— ¹ cl— c—R ₇ R6 (CH ₂ ), • ^b a

O x<CH ₂ ) _in ~x || ( γ Cl—C—N

Rs cl—C—N ^ ^B 2 b ₃ í? ⁰

11

Cl—c—CH ₂ —O— R10 Cl—C—NH—SO ₂ —R ₇ o Rs R ₉ oo

II I I II II

Cl-c-Ν—A ₂ —N—c—R ₄ Cl—C—alkyl-SO ₂ —R7

O R9 ř íř _p_Ň__ _and „_N—C— OR4 or CI-SO2-R7,

Cl—C—aryl—SO ₂ —R7 Cl-CN—A ₂ or by reaction with OCN-SO2-R9 the compound of formula (XV) is prepared

N-Pi

HN-CN—(CH ₂ ) _q Pl

-Rl5

N

0'

Xi

Removal of the N-protecting groups then provides compounds of formulae IV, V and VI.

Compounds of formulae IV and VI in which R1 or R14 is carboxy or alkoxycarbonyl can be prepared by reacting an intermediate of formula XIII with an alcohol, bromide or iodide of formula (XVI) HO-Z, Br-Z or I-Z where Z is a group including alkyl, substituted alkyl, benzyl or benzhydryl. When compound XVI is HO-Z, the reaction is carried out in the presence of a coupling agent such as dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylamino)propylcarbodiimide, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, or carbonyldiimidazole. When compound XVI is Br-Z or I-Z, the reaction is carried out in the presence of a base such as sodium carbonate or sodium bicarbonate. The reaction prepares a compound of formula (XVII)

N-Px

II

HN— CN— (CH ₂ ) _q ki .CO ₂ Z

-NH • · · · • · ♦ ···« · · · ·

By reacting a selected intermediate of formula XVII with an acid chloride of a group comprising

O

Cl—C—CH ₂ -O— Rio Cl—C—NH SO ₂ —R7

About Rg

Rg O I

About Rg

Rg OI II cl- c- N—A ₂ —N— c—R ₄ Cl- c- N—A ₂ —N— C~ OR ₄

About About

II or II _D

Cl—SO ₂ -R ₇ cl—C—alkyl-so ₂ —R7 Cl—C—aryl—SO ₂ or by reaction with OCN-SO2-R7, the compound of formula (XVIII) is prepared

When Z represents a protecting group such as benzyl or benzhydryl, removal of this group and the N-protecting groups from the compound of formula XVIII provides the desired compounds of formulae IV and VI in which R 1 or R 14 represents a carboxy group.

Also in the case where Z represents an alkyl group and Xi represents O—C—NH—R4, it is possible to treat the compound of formula XVIII to remove the N-protecting groups followed by mild hydrolysis in aqueous medium to prepare the desired compounds of formulas IV and VI in which Ri or R14 represents a carboxy group.

means

Compounds of formula III can be prepared by treating a carboxy-substituted azetidinone of formula IX with a haloalkyl of formula (XIX) halogen-R12 in the presence of a base, where halogen is Cl, Br, or I, to prepare an azetidinone of formula (XX)

Cl—(CH ₂ ) _q -

-1 1-]

every ₂ hours

O

The azetidinone of formula XX can also be prepared by reacting the azetidinone of formula VII with a haloalkyl of formula XIX in the presence of a base to prepare the azetidinone of formula

HS—np ₃

By treating a compound of formula XXI with an alkyl dihalide (xxi) ^:< 12 ^H CO ₂ H • · of formula VIII in the presence of a base, an azetidinone of formula is prepared.

XX.

The azetidinone of formula XX is then reacted in the same manner as the azetidinone of formula IX as described above to yield the desired compounds of formula III.

Compounds of formula II in which E1 is other than alkyl can be prepared by treating an olefin of formula

Rn(XXII)I

HC=c—CH=CH ₂ with chlorosulfonyl isocyanate to obtain the azetidinone of formula (XXIII)

Oxidation of compound XXIII, for example by treatment with potassium permanganate, followed by silylation, for example with tri(lower alkyl)silyl chloride, produces the azetidinone of formula (XXIV)

Rll

_CO2H

N—P ₃

The azetidinone of formula XXIV is then reacted in the same manner as the azetidinone of formula VII as described above to prepare the desired compounds of formula II.

Compounds of formula II, in which Eii and Ei both represent an alkyl group, can be prepared by treating an azetidinone of formula (XXV)

Ru

-Ri j—np ₃ where R1 and En both represent an alkyl group, in the same manner as the azetidinone of formula VII as set forth above. Dialkyl-substituted azetidinones of formula XXV are known in the art, see, for example, US Patent 4,775,670.

Compounds of formulae IV and VI in which R1 or R14 is -(CH2)2~ can be prepared by reacting an N-protected guanidine of general formula (XXVI)

N-Px

Ρχ— NC-NH— (CH ₂ ) _q — CH ₂ —CO ₂ CH ₃ with N-protected compound of general formula (XXVII)

P ₂ -N=CH-CH=CH-aryl where P2 represents a trimethylsilyl group, in the presence of lithium diisopropylamide, to obtain an azetidinone of the general formula

Pl— N- C-NH— (CH ₂ )

H

CH=CH—aryl

-NH

About (XXVIII)

N-Px is then reacted with groups including

An azetidinone of formula XXVIII with an acid chloride selected from

O

II

Cl—C— CH ₂ - O— Rio

O

II cl— C—NH—so ₂ —R?

About Rg

II I

Rg O l II

Cl-CN—A ₂ —N—c—R ₄

O

II

Cl— C—alkyl—SO ₂ — ^R 7 o

II

Cl— C—aryl—SO ₂ — R7

WED ⁹ T ⁹

Cl-CN—A ₂ —N—

O

II

C-OR4 or with CI-SO2-R7, or with OCN-SO2-R7, and thus, after subsequent reduction of the alkene group and removal of the protecting groups Pi, the desired compounds of formulas IV and VI are obtained.

Compounds of formulae II, IV and VI can also be prepared by reacting an acid chloride of general formula (XXIX)

Cl—(CH ₂ ) _q — C— Cl with N-protected compound of general formula ΐ ²

H ₃ C- O—ζθ^>— N= C— COOCH3 in the presence of a base, thereby preparing an azetidinone of general (XXX) • · · formula (XXXI)

Treatment of the azetidinone of formula XXXI with sodium azide affords the azetidinone of general azide, such as that of formula (XXXII)

N ₃ —(CH ₂ )

-cooch ₃

OCH ₃

Treatment of the compound of formula XXXII with ceric ammonium sulfate removes the methoxyphenyl group and the resulting azetidinone can be subjected to the same reaction as the azetidinone of formula X as described above to provide the desired compounds of formulas II, IV and VI.

Compounds of general formula I in which Ai is (CH ₂ ) / 1^2/ t

(CH ₂ ) _u — can be prepared by reacting a compound of the general formula (CH ₂ ) _t (CH ₂ ) _u -Li (XXXIII) where Li represents a leaving group such as bromine or iodine, with an azetidinone of the formula VII, XXI, XXIV or XXV in the presence of a base, whereby the reaction produces an azetidinone of the general formula (XXXIV) (CH ₂ )t (CH ₂ )

COOH

Removal of the protecting group Pi and treatment of the above azetidinone as described above for the azetidinone of formula XI provides the desired compounds of formula I. Alternatively, the azetidinone of formula XXXIV can be first treated with an acid chloride to introduce the desired group Xi, deprotected, and reacted with a guanylating agent of formula XII.

Alternatively, compounds of general formula I in which Ai is ✓ (CH ₂ ) _t ''(CH siu

Ri represents a -(CH2)2-aryl group and R2 and R3 represent hydrogen, prepared by reacting a compound of the general formula ( _CH2 ) _{U- CH2} - _COOCH3 (XXXV)

with a reagent of general formula XXVII in the presence of 1 lithiumdiisopropylamide to form an azetidinone of general formula (XXXVI)

Pl— (CH ₂ )

(CH ₂ ) _u •CH= CH-aryl

NH

Removal of the protecting group Pi and reaction with a guanylating agent of formula XII prepares the azetidinone of formula (XXXVII) NP _x ( _C H ₂ ) _t

HN-C—tt >

¹ ^XcH ) (CH ₂ ) _U

CH=CH-aryl

-NH

By reacting an intermediate of general formula XXXVII with an acid chloride selected from the group consisting of ci—c—r ₇

O ^<CH ₂ ) _v ·^

Cl—c—N ^Y (ch ₂ )·/

About II

-(CH ₂ ) _m i? Π — CN-1— ¹

Cl—C N-|

Cl-C—

Rs.

Re( _CH2 ) _o ( _CH2 )n

Rq O II *Bi •Re z^> ^B 2 b ₃ d—C CH ₂ -O—Rio Cl—CN—A ₂ —N—C—R* / ' oo _Ί II II

Cl C alkyl—SO ₂ —R7 Cl—C—aryl—SO ₂ —R7 >

ř ί ⁹ ? ř cl- c- N—A ₂ —N— c— OR ₄ or with CI-SO2-R7, or reaction with OCN-SO2-R7, and subsequent reduction of the alkene group and removal of the protecting group Pi, the desired compounds of formula I are obtained.

Alternatively, compounds of formula I in which Ai is (CH ₂ ) are possible.

(CH ₂ ) _u prepared from azetidinone of formula XXXVII. According to this method, a protecting group is introduced onto the ring nitrogen atom of the azetidinone of formula XXXVII, for example by treatment with tert-butyldimethylsilyl chloride. Treatment with ozone results in reduction of the group

-CH=CH to the aldehyde, which is then converted by Jones oxidation or treatment with sodium chlorite and amidosulfuric acid to the carboxylic acid. Removal of the silyl protecting group gives the azetidinone of the formula

(XXXVIII)

P _X N—C—

(ch ₂ ) _u -j-COOH

-NH

Treatment of the above azetidinone in the manner described for the azetidinone of formula XIII affords the desired compounds of formula I.

• · » · * · « · · > · · ·«·· ·«

The following method is a preferred method for preparing the intermediate of formula XIII. According to this method, a silyl-protected azetidinone of formula VII is treated with an N-protected iodine derivative of the general formula

Pi (XXXIX) X (CH ₂ )qI

Pl whereby, after subsequent removal of the silyl protecting group, an azetidinone of the general formula < (XL) is obtained

Pi \

pf

N-(CH ₂ )q,CO ₂ H

NH which can be isolated in the form of an amine salt, tert-butylamine salt.

for example

The protecting groups Pi are then removed from the azetidinone of formula XV and the resulting compound is reacted with a guanylating agent with two introduced protecting groups of formula XII to give the intermediate of formula XIII, which can be isolated again in the form of the tert-butylamine salt.

The iodine derivative of formula XXXIX can be prepared by reacting a doubly protected amine of the general formula

Pi (XLI) ^NH with an alkyl dihalide of formula VIII to prepare a chlorine derivative of formula (XLII)

Pi/N-( _CH2 ) _q -Cl

Pl

9 9 9 9 ·· 99 • * «999 *·

The chlorine derivative of formula XLII is then treated with sodium iodide in the presence of base to give the iodine derivative of formula XXXIX.

The alternative method below can also be used to prepare compounds of formulae IV and VI.

The reaction of the azetidinone of formula IX with benzyl chloroformate in the presence of triethylamine and dimethylaminopyridine produces the benzyl ester of general formula (XLIII)

C1-(CH ₂ )

By treating the chlorine derivative XLIII with sodium iodide, an iodine derivative of the general formula (XLIV) is obtained.

I-(CH ₂ ) _q ,CO ₂ -CH ₂

0'

-NP ₃

By reacting the above iodine derivative of formula XLIV with guanidine with two introduced protecting groups of formula (XLV)

N—Pi

Pl—

H prepares an azetidinone derivative of formula (XLVI)

N—Pi

H ₂ N—CN—(CH ₂ ) _q ^P 1 ,co ₂ -ch ₂ —(5>

-NP ₃

Removal of the silyl protecting group P3 from the azetidinone of formula XLVI, for example by reaction with ammonium fluoride, gives the azetidine derivative of formula (XLVII)

N-Px

H ₂ N—CN—(CH ₂ ) _q Pi ,CO ₂

-NH

0'

-ch ₂ -<5>

By reacting the intermediate of formula XLVII with an acid chloride selected from the group consisting of

O

II ci— c—r ₇ (CH ₂ ) _o ^ ^b 1, □( CH ₂ ) m

CJ-— V--« 1

Re

Cl-C—N

Cl-C— •Rs

Y( _CH2 ).

(CH ₂ )n ^ ^B 2 'B ₃

Cl—Č—CH ₂ —O— R10 Cl-CN—A ₂ —Ç—c—R ₄ z ' oo

II II

Cl—C—alkyl—S0 ₂ —R7 Cl—C—aryl—S0 ₂ —R7 o R9 Rg OO

II II 11 II cl- cN—A ₂ — N— c— OR4 cl- C-NH-SO ₂ —R ₇ or CI-SO2-R7, or by reaction with OCN-SO2-R7, a compound of the general formula is prepared • · (XLVIII)

N-Ri

H ₂ N—CN—(CH ₂ ) Pi

Removal of the benzyl protecting group and the N-protecting groups Pi from the azetidinone of formula XLVIII gives the desired compounds of formulas IV and VI. —N (CH ₂ ) _q

Compounds of formula I in which A1 is ²⁰ can be prepared by reacting an azetidinone of formula XV with an alcohol, bromide or iodide of formula XVI to prepare an azetidinone of formula (XLIX)

Pi

N-(CH ₂ ) _ _/ CO ₂ Z

J-NH

O

By reacting the intermediate of formula XLIX with an acid chloride selected from the group consisting of

O

II ci— c—r ₇ o

II

Cl—c-(CH ₂ ) _m

N-|—

Re

O

O

II

Cl—C—CH ₂ —O—Rio cl—C NH SO ₂ —R ₇

O R9 Rg O O

II II II II cl- c- N—A ₂ — N— c—R ₄ Cl— C—alkyl—SO ₂ —R7

O

II

Cl— C—aryl—SO ₂ — R7

R9 Rg O

I I II

N—A ₂ — N— c— OR ₄

O

II

Cl-C39 • · • · · • · * · · · • · · · · · · · · · · · · · or CI-SO2-R7, or by reaction with OCN-SO2-R9, a compound of formula (L) is prepared

Pi zCO ₂ Z

Pl

-(CH ₂ )q—

-Ns 'Xi

Removal of the protecting groups Pi, such as by treatment with trifluoroacetic acid in the case where Pi represents a tert-butoxycarbonyl group, produces the amine salt with trifluoroacetic acid of formula (LI)

HOOC-CF3 · H ₂ N (CH ₂ )

_CO2Z

-N.

Xi

Treatment of the compound of formula LI in the form of its amino salt with trifluoroacetic acid with a suitable aldehyde in the presence of a reducing agent such as sodium triacetoxyborate or cyanotrihydroborate affords the compound of formula (Lil) >co ₂ z

R ₂ o-NH (CH ₂ ) q—-(

Xi

Reaction of a compound of formula L1 with a guanylating agent of formula XII with two introduced protecting groups prepares an azetidinone of formula • fl • flflfl (L1)

Pχ-N—C—N (CH ₂ )R20

O

Removal of the protecting groups Pi and Z gives the desired compounds of formula I.

The azetidinone derivatives of formulas I to VI and various intermediates and starting materials used in their preparation contain one or two asymmetric carbons in the formula below shown at positions 3 and 4.

Of course, compounds of formula II in which R1 and R11 have the same meaning and compounds of formula VI in which R14 is hydrogen contain only one ring asymmetric carbon atom. Additional asymmetric carbon atoms may be present in compounds of formula I to VI depending on the meanings of the substituents R1, A1, Xi, X2, R13, X3 and R14. As is well known in the art, see for example J. March, Advanced Organic Chemistry, 4th edition, John Wiley & Sons, New York, NY (1991) pp. 94-164, said asymmetric carbon atoms allow the formation of enantiomers and diastereomers, and all such diastereomers, both in pure form and in the form of mixtures thereof, are included in the invention. Furthermore, if compounds of formula I to VI contain alkenes, they may, if substituted in an appropriate manner, exist as cis- or trans- isomers or mixtures thereof. All such forms are again within the scope of the invention.

The compounds of formulae I to VI can be prepared in the form of their pharmaceutically acceptable salts, their physiologically hydrolysable esters or in the form of their solvates. The compounds of formulae I to IV and VI, in which R1 or R2 represent a carboxy group, can exist in the form of an inner salt or in a zwitterionic form. All of the above forms are encompassed by the invention. Said pharmaceutically acceptable salts include salts with mineral acids such as hydrochloric, hydrobromic, phosphoric and sulphuric acid and also salts with organic carboxylic acids or sulphonic acids such as acetic, trifluoroacetic, citric, maleic, oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic, malic, methanesulphonic, p-toluenesulphonic and the like. The preparation of said acid addition salts is carried out by conventional methods.

The novel compounds of formulae I to V and the compound of formula IV exhibit tryptase inhibitory activity. This activity was confirmed on both isolated human skin tryptase and on recombinant human tryptase prepared from human recombinant beta-protryptase expressed by baculovirus in insect cells. The expressed beta-protryptase is purified by successive use of an affinity resin with an anchored heparin and an immunoaffinity column using an anti-tryptase monoclonal antibody. The protryptase is activated by autocatalytic removal of the N-terminal in the presence of dextran sulfate followed by removal of two N-terminal amino acids by dipeptidyl peptidase I (DPPI) to form the final active enzyme (Sakai et al., J.Clin.Ivest., 97, pp.988-995, 1996). Essentially equivalent results were obtained using the isolated native enzyme and the activated expressed enzyme. The above tryptase is stored in 2 M sodium chloride, 10 nM 4-morpholinepropanesulfonic acid pH

6.8.

The assay is performed in a 96-well microplate. 250 μΐ of assay buffer (containing low molecular weight heparin and tris(hydroxymethyl)aminomethane) is added to each well of the microplate (Nunc MaxiSorp) followed by 2.0 μΐ of the test compound in dimethyl sulfoxide. Substrate (10 μΐ) is then added to each well such that the final mixture contains either 370 μΜ benzoyl-arginine-p-nitroaniline (ΒΑΡΝΑ) or 100 μΜ benzyloxycarbonyl-glycine-proline-arginine-p-nitroaniline (CBz-Gly-Pro-Arg-pNA). Similar results were obtained using both substrates. The plate is then placed on the plate of a vortex mixer set at 800 (Sarstedt TPM-2). After a total of three minutes of incubation, 10 μΐ of tryptase working solution is added to each well (final tryptase concentration in case of using ΒΑΡΝΑ 6.1 mM and in case of using

Cbz-Gly-Pro-Arg-pNa 0.74 nM). The plate is then vortexed again for one minute and then incubated without shaking at room temperature for another 2 minutes. The plate is then evaluated for twenty minutes at room temperature using a plate reader (Molecular Devices UV max) in kinetic mode (wavelength 405 nm). To determine the concentration of compound that inhibits the enzyme activity by one half (IC50), the activity of the samples with affected activity (FCA) is plotted against the concentration of inhibitor (I) and the curve fitting the FCA/(1+[I]/IC50) curve is evaluated. The IC50 value is determined for each compound 2 to 4 times and the average value is calculated from the values obtained.

Due to the said activity against tryptase, the compounds of formula I to VI, as well as their inner salts, their pharmaceutically acceptable salts, their hydrolyzable esters and solvates, are suitable as anti-inflammatory agents, in particular in the treatment of chronic asthma and can also be used in the treatment or prevention of allergic rhinitis, inflammatory bowel disease, psoriasis, conjunctivitis, atopic dermatitis, rheumatoid arthritis, osteoarthritis and other chronic inflammatory joint diseases or diseases of joints with cartilage destruction. In addition, the said compounds are suitable in the treatment or prevention of myocardial infarction, stroke, angina and other consequences of rupture of atherosclerotic plaque. In addition, the said compounds may be suitable for the treatment or prevention of diabetic retinopathy, tumor growth and consequences of angiogenesis. Furthermore, the said compounds may be suitable for the treatment or prevention of fibrotic conditions, for example fibrosis, scleroderma, pulmonary fibrosis, liver cirrhosis, myocardial fibrosis, neurofibromas and hypertrophic scars.

Compounds of formula I to IV are also inhibitors of factor Xa and/or VIIa. Therefore, the compounds of formulae I to VI, as well as their inner salts, pharmaceutically acceptable salts, hydrolysable esters or solvates, may be useful in the treatment or prevention of thrombotic events associated with coronary arteries and cerebrovascular disorders, including the formation and/or rupture of atherosclerotic plaques, venous or arterial thrombosis, coagulation syndromes, ischemia and angina (stable and unstable), deep vein thrombosis (DVT), disseminated intravascular coagulopathy, pulmonary embolism, myocardial infarction, cerebral infarction, transient ischemic attack, atrial fibrillation, cerebral embolism, thromboembolic complications of surgery (such as hip or knee replacements, heart valves and endarterectomy) and occlusion of peripheral arteries, and may also be useful in the treatment or prevention of myocardial infarction, stroke, angina and other consequences of atherosclerotic plaque rupture. Compounds of formulae I to VI exhibiting inhibitory activity against factor Xa and/or factor VIIa may also be useful as inhibitors of blood clotting, for example in the collection, storage and fractionation of whole blood.

The compounds of formulae I to VI are also inhibitors of urokinase plasminogen activators. As a result, the compounds of formulae I to VI, as well as their inner salts or pharmaceutically acceptable salts, their hydrolysable esters or solvates, may be useful in the treatment or prevention of restenosis and aneurysms, in the treatment or prevention of myocardial infarction, stroke, angina and other consequences of rupture of atherosclerotic plaque, and may also be useful in the treatment of malignant diseases, for the prevention of the spread of metastases, for the prevention of prothrombotic complications of malignant tumours and as an adjunct to chemotherapy.

Compounds of Formulae I to V also have thrombin and trypsin inhibitory activity similar to that described by Han in the U.S. patent referenced above for compounds of Formula VI. Accordingly, compounds of Formulae I to V, as well as their inner salts or pharmaceutically acceptable salts, hydrolysable esters or solvates thereof, may be useful in the treatment or prevention of pancreatitis, in the treatment or prevention of thrombotic events associated with coronary arteries and cerebrovascular disorders as noted above, and may also be useful as inhibitors of blood coagulation during collection, storage and fractionation of whole blood.

Certain compounds of formulae I to IV are also useful in view of their selective inhibitory activity against tryptase. These compounds which have high inhibitory activity against tryptase are much less active against other enzyme systems including trypsin, thrombin and factor Xa. This selective activity on tryptase is evident in compounds of formulae I to IV, in which X1 or X2 represents a group

^R 25 and R 25 is a linking intermediate group terminated by a lipophilic group. Suitable linking intermediate groups include groups of 5 or more atoms such as

-CH ₂ ) ₅ -io-, -O-(CH ₂ )4-9-, —NH—(CH 2 )4 - 9—, — (CH ₂ )4 - 9—0—, -O-(CH ₂ )3-8-0-, —N—(CH ₂ )4 - 9—, -NH-(CH ₂ )3-8-0-, and

CH ₃

-N-(CH2)3-8-0-, etc.,

CH ₃ also as groups containing 3 or more atoms and a phenyl ring such as

-NH—(CH ₂ ) ₂ —O—(CH ₂ ) ₂

ch ₃ a

etc.

Suitable lipophilic terminal groups include those selected from the group consisting of aryl, substituted aryl, heteroaryl, aryl-A3-aryl, aryl-A3-substituted aryl, aryl-A3-substituted aryl, and aryl-A3-heteroaryl, etc. The compounds of formulae I to IV, as well as their inner salts and pharmaceutically acceptable salts, hydrolysable esters or solvates thereof, are useful as anti-inflammatory agents, particularly in the treatment of chronic asthma, and may be useful in the treatment or prevention of allergic rhinitis as well as other diseases listed above for non-selective tryptase inhibitors. It is believed that the selective inhibition of tryptase by these compounds results in fewer undesirable side effects.

The compounds of formulae I to VI, as well as their inner salts or pharmaceutically acceptable salts, their hydrolysable esters or solvates, can be administered orally, topically, rectally or parenterally, or they can be administered by inhalation into the bronchioles or nasal passages. The route of administration will of course vary depending on the disease to be treated. The amount of active compound administered will also vary depending on the route of administration and the disease to be treated.

An effective amount is in the range of about 0.1 to about 100 mg/kg, preferably in the range of about 0.2 to about 50 mg/kg, and more preferably in the range of about 0.5 to about 25 mg/kg/day in a single dose or in multiple doses administered at appropriate intervals throughout the day.

The composition used in the above-mentioned therapies can have various forms. Said forms include, for example, solid, semi-solid and liquid dosage forms such as tablets, pills, powders, liquid solutions or suspensions, liposomes, injection and infusion solutions. Said compositions can contain pharmaceutically acceptable carriers, preservatives, stabilizers and other agents commonly used in the pharmaceutical industry.

If the compounds of formulae I to VI, as well as their inner salts or pharmaceutically acceptable salts, their hydrolysable esters or solvates are to be used for the treatment of asthma or allergic rhinitis, it is advantageous to formulate them in the form of an aerosol. The term aerosol includes any phase of the active compound suspended in a carrier gas allowing inhalation into the bronchioles or nasal passages. Aerosol preparations comprise a suspension of droplets of the active compound in a carrier gas which is formed in a metered dose inhaler, nebuliser or air-jet nebuliser. Aerosol preparations also include dry powder compositions which are suspended in air or another carrier gas. The concentration of the active compounds of formulae I to VI in the solution used to prepare the aerosol preparations is from about 0.1 to about 100 mg/ml, more preferably 0.1 to about 30 mg/ml and most preferably from about 1 to about 10 mg/ml. Said solution typically contains a pharmaceutically acceptable buffer such as a phosphate or bicarbonate buffer suitable for adjusting the pH to about 5 to 9, preferably 6.5 to 7.8, and most preferably 7.0 to 7.6. As is consistent with conventional pharmaceutical practice, said preparations may contain preservatives and other agents.

In combination with the compounds of formulae I to VI, other pharmaceutically active agents may be used depending on the disease to be treated. For example, in the treatment of asthma, β-adrenergic agonists such as albuterol, terbutaline, formoterol, fenoterol or prenaline may be used, as may anticholinergic agents such as ipratropium bromide, anti-inflammatory corticosteroids such as beclomethasone, triamcinolone, flurisolide or dexamethasone, and anti-inflammatory agents such as cromolyn and nedocromil.

In addition to the novel compounds of formulas I to V and methods of using compounds I to VI, the invention includes novel intermediates and novel synthetic methods for preparing these compounds.

Preferred compounds of the invention are those compounds of general formula IV in which:

q means 3;

R1 represents a carboxy group:

O _n O ___ O ,-<

n /—\ ¹¹ / \ ^{11 yZ} ίη,.·.--γ2>-Κ< -C-γ^ΝΗ or — C—NH—(substituted alkyl);

X ₂ means

-cn 7 ÍI \_/ — cT ⁹ ř

N—(CH ₂ ) ₂ —N—C— OR _{4 ;} or ír T ⁹ T ⁹ ír

CN—(CH ₂ ) ₂ —N—C—R ₄ ·

R e represents aminocarbons 1, ^c NH(alkyl), or -c—N(alkyl)

Y means NR ₄ , íř ř

N—C—A ₃ R ₇ , N—C—A ₃ -0-R ₇ íir íir ií

N ^- C—O—A ₃ —R ₇ N—C—A ₃ -CR ₇ or

F~\

- ^R 4

R ₄ within the meaning of Y and X ₂ represents a group including alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, and heteroaryl;

R7 represents a group from the group consisting of alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, ··· ·»*··«· • · · ’ · ·· · aryl, -(CH2)1-4-aryl, -(CH2)1-«-aryl-A3-aryl,

A — Ν _χ neb» _CH ₂ -N _X ^r 5 _t ^Rs ;

A represents an amino group, -NH(alkyl) or R5

-(alkyl)2;

R9 represents a lower alkyl group;

A3 means a bond, an alkylene bridge of 1 to 6 carbon atoms, — (CH ₂ ) _d -N-(CH ₂ ) _e —

R21 or — (CH ₂ ) _d —O-(CH ₂ ) _e — .

d and e are independently selected from the group consisting of zero and an integer from 1 to 6;

R21 represents hydrogen or a lower alkyl group;

and their internal salt when edible salt.

or their pharmaceutical

Also preferred are those formulae I in which

Ai means compounds according to the invention of general

R2 and R3 both represent hydrogen;

R1 has the meaning given above;

Xi means

or

—CN-(CH ₂ ) 2 -NeR* where Y, R 4 and R 9 preferably have the meaning given above;

t represents 2 or 3;

u means one;

and an inner salt or a pharmaceutically acceptable salt thereof.

Most preferred are the following compounds of general formula IV including their inner salt or pharmaceutically acceptable salt form:

every ₂ hours

O

II

Kr Nc-( _CH3 )3 < H

0' • · *···« · • » · «·* · ··

c—iZ — c—nh ₂

Νχ /jZ π—C—O—CH—CH(CH ₃ ) ₂

S Á_/ n ¹ o ⁰ CH(CH ₃ ) ₂

/—\ u

Νχ / _N N— CO-CH-CH(CH ₃ ) ₂ £ v_y lo CH(CH ₃ ) ₂

The most preferred compounds are also the following compounds of general formula I including their inner salt or pharmaceutically acceptable salt form:

• · • · · · · • · « • · ’

I · · · · ·

O

N- c- O- CH—CH (CH ₃ ) 2 CH(CH ₃ ) ₂

The following compounds of general formula IV, including their inner salt or pharmaceutically acceptable salt form, are also preferred:

• · · *

NH χ

H ₂ NN' H _k CO ₂ H •yj

NC— CH ₂ -N[CH(CH ₃ ) ₂ ] ₂

NH

X

H ₂ NN' H >CO ₂ H

N\ N ? and about

o o II II

NCCN-CH—CH (CH ₃ ) 2

CH(CH ₃ ) ₂

NH

X

H ₂ NN' H _to CO ₂ H

-N. N—(CH ₂ ) ₂ —Ν' in

AND._

O—C(CH ₃ ) ₃ ch ₃ ch ₃

NH h ₂ n^n^^ ^/z <

H

_CO2Hn

CH ₃ ch ₃ \ II fi CN-(CH ₂ ) ₂ -N—c0 ch ₃ ch ₃ ch ₃ •ch ₃

NH

X h ₂ nn' H

Tf _to CO ₂ H

O

1_/ \ II

NN— c- o- CH- CH ₂ -CH (CH ₃ ) ₂

II

O

_CH2 -CH( _CH3 ) ₂

NH

Η ₂ Ν^Ν ^/ ^ ^χ/// ''' ² H kCO ₂ H

• * • · to · · · · ·

O

II

NH

AT

H ₂ NN' H

Hk Ν N— C—CH ₂ —O— CH—CH(CH ₃ ) ₂

CH(CH ₃ ) ₂ c-nh-ch ₂ -c-nh ₂

0' /-\ u

BL /N NC—0—CH—CH(CH ₃ ) ₂ c \_y 1 “ CH(CH ₃ ) ₂

To illustrate the invention, examples of its embodiments are given below.

Examples of embodiments of the invention

Example 1

and)

(homochiral)

N ₃

>C0 ₂ H

F——NH O

To a solution of diisopropylamine (13.5 mL, 95.9 mmol) in dry tetrahydrofuran (40 mL) under nitrogen at -78 °C with mechanical stirring was added dropwise over 5 min butyllithium in hexanes (2.5 M, 36.6 mL, 91.5 mmol). The mixture was then warmed to 0 °C, stirred for 30 min, then cooled again to -78 °C and (4S)-N-(tert-butyldimethyl)-55 was added in one portion.

silyl)-azetidin-2-one-4-carboxylic acid (10 g, 43.6 mmol) (Baldwin et al., Tetrahedron, Vol. 46, pp. 4733-4748). The reaction mixture, which is a gel-like suspension, is then stirred for 5 minutes at -78 °C, then the reaction mixture is warmed to -20 °C to -10 °C and stirred at this temperature for 30 minutes. Then 1-chloro-3-iodopropane (5.7 ml, 53.0 mmol) is added in one portion and the reaction mixture is stirred for 2 hours at -20 °C (the gel-like suspension disappears upon addition of 1-chloro-3-iodopropane). The reaction mixture was then poured into 1N HCl saturated with sodium chloride (300 mL) and the aqueous phase was extracted with ethyl acetate (1 x 150 mL), which was then washed twice with saturated 1N HCl. The aqueous layers were then extracted twice, sequentially, with ethyl acetate (2 x 150 mL). The combined organics were then extracted twice with water at pH 7.5-8 (2 x 100 mL, adjusted by dropwise addition of 25% sodium hydroxide). The combined basic aqueous portions were then washed with ethyl acetate (2 x 150 mL). The basic aqueous portions were then acidified with concentrated HCl to pH 3, saturated with sodium chloride (solid) and extracted with ethyl acetate (2 x 150 mL), which was dried over sodium sulfate, filtered and concentrated. Evaporation with toluene then gave the compound

as a crude yellow oil. TLC (silica gel, acetic acid in ethyl acetate) Rf=0.1, bands.

1%

To the crude chloride solution prepared above (less than

43.6 mmol) in dry dimethylformamide (40 ml) under nitrogen atmosphere were added tetrabutylammonium iodide (0.5 g, 1.36 mmol) and tetrabutylammonium azide (15 g, 52.7 mmol). The reaction mixture was then stirred for 72 hours at room temperature, after which the major portion of • · · · · · ♦ >

• · dimethylformamide is removed in vacuo. The product is extracted into ethyl acetate (150 mL), which is washed with 1 N HCl saturated with sodium chloride (3 x 150 mL). The aqueous layers are extracted twice in succession with ethyl acetate (2 x 150 mL). The combined organics are then extracted into water at pH 7.5-8 (2 x 100 mL, adjusted by adding 25% sodium hydroxide dropwise). The combined basic aqueous layers are then washed with ethyl acetate (3 x 150 mL). The basic aqueous layers are then acidified with concentrated HCl to pH 3, saturated with sodium chloride (solid) and extracted with ethyl acetate, dried over sodium sulfate, filtered and concentrated. Evaporation and drying with toluene afford the desired azide (6.56 g, 33.1 mmol) as a tan foam. TLC (silica gel, 0.1% acetic acid in ethyl acetate) Rf=0.1.

To a solution of the azide prepared in step (a) (6.5 g, 32.8 mmol) in dimethylformamide (40 ml) was added acetic acid (4 ml, 66 mmol) followed by 10% palladium on carbon (1.3 g). Hydrogen was then bubbled through the reaction mixture for 25 minutes and the reaction mixture was stirred for 6 hours under a hydrogen atmosphere. After degassing the mixture with nitrogen for 25 minutes, triethylamine (15 ml, 108 mmol) and N,N'-bis(benzyloxycarbonyl)-1-guanylpyrazole (15 g,

39.7 mmol) (Wu et al., Synthetic Communications, 23(21) pp.3055-3060, (1993). The reaction mixture was then stirred for 18 hours at room temperature. The reaction mixture was then filtered through Celite ^R and the pad was washed with ethyl acetate. The solvents were removed in vacuo and the residue was dissolved in ethyl acetate.

(150 mL) and washed with 1 N HCl saturated with sodium chloride (3 x 150 mL). The aqueous washings were successively extracted with ethyl acetate (2 x 100 mL). The combined organics were then extracted with water at pH 7.5-8 (2 x 150 mL, adjusted by adding 25% sodium hydroxide dropwise). The combined basic aqueous layers were then washed with ethyl acetate (3 x 150 mL) to remove excess Ν,Ν'-bis(benzyloxycarbonyl)-1-guanylpyrazole from the product. The basic aqueous layers were then acidified with concentrated KCl to pH 3, saturated with sodium chloride (solid), extracted with ethyl acetate (2 x 150 mL), dried with sodium sulfate, filtered, and concentrated. Evaporation drying with toluene gave a light brown foam. Purification by flash chromatography (silica gel, 1-3% acetic acid in ethyl acetate) gave the desired product (8.5 g, 17.6 mmol) as an off-white solid. TLC (silica gel, 1% acetic acid in ethyl acetate) Rf=0.2.

O

To a solution of the product prepared in step (b) (1.7 g,

3.52 mmol) in dimethylformamide (20 ml) at room temperature under nitrogen atmosphere was added solid sodium bicarbonate (1.5 g, 17.9 mmol), tetrabutylammonium iodide (200 mg, 0.54 mmol) and finally benzyl bromide (2.5 ml, 21.0 mmol). The reaction mixture was stirred for 48 hours at room temperature. Then the dimethylformamide was removed in vacuo and the residue obtained was dissolved in ethyl acetate and the ethyl acetate was then washed twice with saturated aqueous sodium bicarbonate. The organic phase was

• · · · separated, dried over magnesium sulfate, filtered, and concentrated to a brown oil. Purification by flash chromatography (silica gel, 0-10% methanol in methylene chloride) gave the desired product (1.84, 3.21 mmol) as a yellow oil.

O

O

To a solution of the product prepared in step (c) (46 mg,

0.08 mmol) in dry tetrahydrofuran (1.0 mL) was added to a solution of bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 100 μΐ, 0.1 mmol) at -78 °C under nitrogen. The reaction mixture was stirred for 10 min at -78 °C and then for 10 min at -20 °C. After cooling the reaction mixture to -78 °C, benzyl isocyanate (100 μΐ, 0.78 mmol) was added in one portion. The reaction mixture was then stirred for 5 min at -78 °C and then for 15 min at -20 °C. 1 N HCl (1 mL) was then added followed immediately by ethyl acetate (3 mL). The resulting biphasic solution was then stirred vigorously while being allowed to warm to room temperature. The organic phase was then separated and washed once with saturated aqueous sodium bicarbonate, dried over magnesium sulfate, filtered, and concentrated to give a light yellow residue. Purification by flash chromatography (silica gel, 0-30% ethyl acetate in hexane) gave the desired product (33 mg,

0.047 mmol).

E)

O

• · · · · • · · · · *

To a solution of the product prepared in step (d) (33 mg, 0.047 mmol) in dioxane (2 mL) was added concentrated HCl (4 μΐ, 0.048 mmol) followed by the catalyst, 10% palladium on carbon (15 mg). Hydrogen was then bubbled through the reaction mixture for 15 min. Water (0.5 mL) was then added and the reaction mixture was stirred under hydrogen for a further 1 h. The reaction mixture was then filtered through Celite ^R and the filter pad was then washed with three portions of water. The combined eluates were then lyophilized to give a white powder. Purification by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) followed by lyophilization gave the desired product (8.7 mg, 0.019 mmol). IR (film) 1773 cm ^-1 ; MS 348.0 (M+H) ⁺ , 346.3 (MH).

Example 2

To a solution of the product prepared according to Example 1(c) (92 mg, 0.161 mmol) in dry tetrahydrofuran (1.5 mL) at room temperature under nitrogen was added sodium hydride (60% in mineral oil, 20 mg, 0.5 mmol). The reaction mixture was stirred for 10 min and then cyclohexyl isocyanate (100 μΐ, 0.78 mmol) was added in one portion. The reaction mixture was then stirred for 30 min at room temperature. The reaction mixture was then slowly poured into ice-cold 1 N HCl (2.5 mL). The resulting solution was extracted with ethyl acetate. The organic phase was washed twice with saturated aqueous sodium bicarbonate and then once with brine. The organic layer was then dried over sodium sulfate, filtered, and concentrated. Purification by flash chromatography (silica gel, 0-10% methanol in methylene chloride) gave the desired product (91 mg, 0.13 mmol). MS 698.1 (M+H) ⁺ , 696.4 (MH) - .

h) h ₂ n

(homochiral)

To a solution of the product prepared in step (a) (91 mg,

0.13 mol) in dioxane (2 ml) was added concentrated HCl (11 μΐ, 0.132 mmol) followed by 10% palladium on carbon catalyst (45 mg). H2 was then bubbled through the reaction mixture for 45 min. Water (1 ml) was then added and the reaction mixture was stirred under nitrogen for a further 45 min. The reaction mixture was then filtered through Celite ^R which was then washed with three portions of water. The combined eluates were then lyophilized to give a white powder. Purification by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave the desired product (28 mg, 0.062 mmol) after lyophilization. IR (KBr) 1773 cm ^-1 ; MS 340.1 (M+H) ⁺ , 338.2 (MH).

·· ·· • · · • * * · • · ⁴ • · • · · · · ·

Example 3

O

The desired product was prepared according to the method of Example 1(d) using 4-biphenylisocyanate instead of benzylisocyanate. IR (film) 1776 cm -1 ; MS 768.1 (M+H) ⁺ , 766.2 (MH).

b) NH»HC1

To a solution of the product prepared in step (a) (30 mg, 0.039 mmol) in dioxane (4 mL) was added concentrated HCl (10 μΐ, 0.12 mmol) followed by 10% palladium on carbon catalyst (30 mg). H ₂ was then bubbled through the reaction mixture for 5 min and the reaction mixture was stirred under hydrogen atmosphere for 1.5 h. The reaction mixture was then filtered through Celite ^R , which was then washed with two portions of water and one portion of dioxane. The combined eluates were then lyophilized to give the desired product (16.3 mg, 0.036 mmol). IR (film) 1769 cm~1; MS 410.1 (M+H) ⁺ , 408.3 (MH)-.

Example 4

The method of Example 1 is followed, except that in step (d) phenethyl isocyanate is used instead of benzyl isocyanate, and the desired product is obtained by deprotection work-up according to the method of Example 1, step (e). IR (film) 1775 cm ^-1 ; MS 362.1 (M+H) ⁺ , 360.3 (MH) ".

Example 5

_H2N

O

H (homochiral)

CF ₃

• · · ·

The method of Example 1 is followed, except that in step (d) 4-trifluoromethylphenyl isocyanate is used instead of benzyl isocyanate, and the desired product is obtained by working up according to the method of Example 1, step (e). IR (film) 1761 cm-1; MS 402.1 (M+H) ⁺ , 400.2 (MH) - .

Example 6

NH«HC1

The procedure of Example 1 is followed, except that in step (d) (R)-α-methylbenzylisocyanate is used instead of benzylisocyanate, and the desired product is obtained by deprotection and work-up according to the method of Example 3, step (b). IR (KBr) 1777 cnr i; MS 362.1 (M+H) ⁺ , 360.2 (MH) -.

Example 7

NH»HC1

(homochiral)

The method of Example 1 is followed, except that in step (d) benzyl isocyanate is replaced by (S)-α-methylbenzyl isocyanate, and the desired product is obtained by deprotection and treatment according to the method of Example 3, step (b). IR (KBr) 1777 cm'i; MS 362.1 (M+H) ⁺ , 360.3 (MH)-.

Example 8 nh«cf ₃ cooh h ₂ n

(homochiral)

The procedure of Example 1 is followed, except that in step (d) 2-biphenyl isocyanate is used instead of benzyl isocyanate, and the desired product is obtained by deprotection by working up according to the method of Example 1, step (e). IR (KBr) 1780 cm-1; MS 410.1 (M+H) ⁺ , 408.2 (MH).

Example 9

The method of Example 1 is followed, except that in step (d) (R)-(-)-(1-naphthyl)ethyl isocyanate is used instead of benzyl isocyanate, and the desired product is obtained by deprotection and processing according to the method of Example 1, step (e). IR (KBr) 1777 cm; MS 412.3 (M+H) ⁺ , 410.2 (MH)

Example 10

The method of Example 1 is followed, except that in step (d) (S)-(+)-(1-naphthyl)ethyl isocyanate is used instead of benzyl isocyanate, and the desired product is obtained by working up according to the method of Example 1, step (e). IR (KBr) 1777 cm; MS 412.3 (M+H) ⁺ , 410.2 (MH) -.

Example 11

a) trans-4-phenylcyclohexyl isocyanate

To a solution of 4-phenylcyclohexanone (5 g, 28.7 mmol) in methanol (150 mL) at room temperature under nitrogen was added ammonium formate (18 g, 285 mmol) followed by sodium cyanohydroborate (1.85 g, 29.4 mmol) portionwise. The reaction mixture was stirred at room temperature for 24 h, then the methanol was removed in vacuo to give an oily residue. The residue was dissolved in methylene chloride (100 mL), and the solution was washed with 1 N sodium hydroxide (2 × 100 mL). The organic layer was then separated, dried over sodium sulfate, and concentrated to give an off-white solid. Purification by flash chromatography (silica gel, 0-20% methanol/methylene chloride) gave 3.46 g of trans-4-phenylcyclohexylamine as a white solid.

To a solution of trans-4-phenylcyclohexylamine (500 mg, 2.85 mmol) in toluene (5 mL) was added phosgene (20% in toluene, 5 mL) at room temperature under nitrogen. The resulting solution was heated at 80 °C for 24 h. The solvents were then removed in vacuo to give a solid residue. The residue was dispersed in ether and the mixture was filtered. The eluted portion was separated and concentrated to give 379 mg of trans-4-phenylcyclohexyl isocyanate as a light yellow oil. IR (film) 2259 cm ^-1 .

b)

The procedure of Example 1 is followed, except that in step (d) trans-4-phenylcyclohexylisocyanate is used instead of benzylisocyanate, and the desired product is obtained by deprotection and work-up according to Example 1(e). IR (KBr) 1778 cm-1; MS 416.2 (M+H) ⁺ , 414.4 (MH).

• · • ·

Example 12

NH»CF ₃ COOH

(homochiral)

and)

0= G=N

To a solution of 3-phenoxybenzaldehyde (5 g, 25.22 mmol) in methanol (125 mL) was added ammonium formate (7.95 g, 126.12 mmol) and sodium cyanohydroborate (4.75 g, 75.66 mmol) and the mixture was stirred overnight at room temperature. After 16 h, the mixture was evaporated in vacuo and partitioned between 1 N HCl and ethyl acetate. The aqueous layer was then basified with 6 N sodium hydroxide solution to pH 12 and re-extracted with ethyl acetate. The organic phase was then washed with brine, dried over sodium sulfate and concentrated to give 300 mg of the product as a colorless oil.

MS 199.2 (M+H) ⁺ .

The above amino compound (300 mg, 1.51 mmol) in toluene (2 mL) was added to a mixture of phosgene (3 mL of a 20% solution of phosgene in toluene) in toluene (2 mL). This mixture was heated at 80 °C for 2 hours, then stirred at 110 °C for 1 hour and then at 80 °C overnight. The mixture was then evaporated in vacuo and the residue suspended in ether and the mixture filtered. The eluted fractions were concentrated and co-evaporated with toluene to give the desired isocyanate as a brown oil (0.328 g). IR (film) 2263.5 cm ^-1 .

• Φ · ·« ♦· • 9

9 9 9

The benzyl ester product prepared according to Example 1(c) (69 mg, 0.121 mmol) was dissolved in tetrahydrofuran (1.5 mL) and cooled to -78°C. Bis(trimethylsilyl)amide (0.15 mL, 0.145 mmol) was then added over 2 minutes and the mixture was stirred at -78°C for 1 hour. A solution of the isocyanate from step (a) (0.328 g, 0.145 mmol) in tetrahydrofuran (1.2 mL) was then added over 1 minute and the reaction mixture was stirred at -78°C. After 30 minutes, the mixture was quenched with 0.5 N sodium hydrogen sulfate (10 mL) and extracted with ethyl acetate (2 x 10 mL). The organic phase was then washed with brine (1 x 15 mL), dried over sodium sulfate and concentrated to give a yellow oil (150 mg). Purification by flash chromatography (silica gel, 0-25% ethyl acetate/hexane) gave the desired product as a light yellow oil (50 mg). MS 798.1 (M+H) ⁺ , 796.3 (MH) ^- ; IR (film) 1776.6 cm ^-1 .

C)

O (homochiral

O'

• 0 0 ·

0« ·· ·» * * c · · • · 0 · · * • 0 ♦ · · *

To a solution of the product prepared in step (b) (45 mg, 0.056 mmol) in 1,4-dioxane (7 ml) containing 1 N HCl was added the catalyst, 10% palladium on carbon (25 mg, wet type). Hydrogen was then bubbled through the solution for 4 hours. The resulting mixture was filtered through Celite ^R , which was then repeatedly washed with 1,4-dioxane. The combined eluates were evaporated in vacuo to give a pale yellow sticky mass (40 mg). Purification by reverse phase preparative HPLC (YMC ODS 30 x 250 mm) using the mobile phase described in Example 1(e) gave the title compound (26 mg) as a white solid. MS 440.2 (M+H) ⁺ , 438.3 (MH) - ; IR (KBr) 1773 cnr i .

Example 13

To a solution of the benzyl ester product prepared according to Example 1(c) (57.9 mg, 0.101 mmol) in dry tetrahydrofuran (2 ml) at -78 °C under nitrogen was added • · · · sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 106 μΐ, 0.106 mmol). The reaction mixture was then stirred for 1 hour at -78 °C and then 1-benzyl-2-phenethyl isocyanate (35 mg, 0.147 mmol), (prepared by the method described in Anderson et al., J.American Pharm.Assoc., Vol.41, pp. 643-650 (1952)) was added in one portion. The reaction mixture was then stirred for 20 minutes at -78 °C. The reaction mixture was then quenched with potassium hydrogen sulfate (3 ml) and then ethyl acetate (5 ml) was immediately added. The resulting biphasic solution was then stirred vigorously while warming to room temperature. The organic phase was separated, dried over sodium sulfate, filtered, and concentrated to give a bright yellow residue. Purification by flash chromatography (silica gel, 0-20% ethyl acetate in hexane) gave the desired product (54.8 mg, 0.068 mmol). IR (film) 1776 curi; MS 810.2 (M+H) ⁺ , 808.4 (MH)'.

(homochiral)

To a solution of the product prepared in step (a) (54.8 mg, 0.068 mmol) in dioxane (3 mL) was added concentrated HCl (20 μΐ, 0.24 mmol) followed by the catalyst, 10% palladium on carbon (50 mg). Hydrogen was bubbled through the reaction mixture for 5 min and then the reaction mixture was stirred for 1.5 h under a hydrogen atmosphere. The reaction mixture was then filtered through Celite ^R , which was then washed with two portions of dioxane and three portions of water. The combined eluates were then lyophilized to give a white powder. Purification after lyophilization by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave the desired product.

product (21 mg). IR (KBr) 1776 cm - 1 MS 452.4 (M+H) ⁺ , 450.4 (MH) - .

a) 1-naphthylmethylisocyanate

A solution of phosgene (20% solution in toluene, 5 ml) was diluted with toluene (10 ml). A mixture of 1-naphthalenemethylamine (500 μΐ, 3.41 mmol) and triethylamine (0.95 ml, 6.82 mmol) was then added dropwise. The reaction mixture was then heated at reflux overnight. The mixture was then cooled to room temperature and the solvent was removed. The residue was stirred with ether (50 ml) for 10 minutes and the mixture was filtered. Concentration of the filtrate gave the crude product, from which the desired product (518 mg) was obtained as a colorless oil by flash chromatography (silica gel, methylene chloride). IR 2260 cm ¹ .

b)

O

To a solution of the benzyl ester product prepared according to Example 1(C) (144 mg, 0.25) in tetrahydrofuran (3 mL) at

At -78 °C, sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 300 μΐ, 0.30 mmol) was added dropwise. The mixture was stirred for 1 h at -78 °C. A solution of 1-naphthylmethylisocyanate (55 mg, 0.30 mmol) in tetrahydrofuran (1 mL) was then added. The reaction mixture was then stirred at -78 °C for a further 40 min. The reaction mixture was then quenched with 1 N potassium hydrogen sulfate (15 mL). The mixture was then extracted with ethyl acetate (2 x 40 mL). The organic layers were combined and washed with brine (15 mL), dried over magnesium sulfate, filtered and concentrated to give 185 mg of crude product as a yellow oil. Purification by chromatography (silica, 30-50% ethyl acetate/hexane) gave the desired product (122 mg) as a colorless oil. MS: (M+H) ⁺ 756.1; IR (KBr) 1776 cm-1, 1732 cm-1, 1639 cm-1.

A mixture of the product prepared in step (b) (117 mg, 0.15 mmol), 1 N HCl (170 μΐ, 0.17 mmol), palladium-on-carbon catalyst (10%, 50 mg) in dioxane (3 ml) was stirred for 1 hour at room temperature under a hydrogen atmosphere (under a hydrogen balloon). The completion of the reaction was confirmed by analytical HPLC. The mixture was then filtered through a pad of Celite ^R and concentrated to give the crude product (68 mg), which was purified by preparative reverse-phase HPLC as described in Example 1(e) to give the desired product (37 mg) as a white powder. MS (M+H) ⁺ 398.2, (MH) - 396.4; IR (KBr) 1780 cm-1, 1670 cm-1, 1541 cm-1.

Example 15 • · • · • · • » • ·

NH«HC1

Λ η ₂ ν ν ² Η £0 ₂ Η

Ύ”ΊΌ (homochiral)

COOCH ₃

The method of Example 1 is followed, except that in step (d) methyl-(S)-(-)-2-isocyanato-3-phenylpropionate is used instead of benzyl isocyanate, and subsequent deprotection and work-up according to Example 3 (b) gives the desired product. IR (KBr) 1769 cm ^-1 , 1674 cm ^-1 and 1632 cm ^-1 ; MS (M+H)+ 420.1.

Example 16

a) R-(+)-1-phenylpropyl isocyanate

A solution of phosgene (20% in toluene, 5 ml) was diluted with toluene (10 ml). Then a mixture of R-(+)-1-phenylpropylamine (640 μΐ, 4.40 mmol) and triethylamine (1.03 ml, 7.4 mmol) in toluene (5 ml) was added dropwise. Then another 10 ml of toluene was added because stirring was difficult. Then the reaction mixture was heated at reflux for 2 hours. TLC analysis showed the end of the reaction. Then the solvent was removed and the residue was stirred with ether (50 ml) for 10 minutes and the mixture was filtered. The filtrate was concentrated to give the crude product, which was purified by flash chromatography (silica, • · • -> · 1 • · · · · β as a colorless oil (410 mg)

2262 cm ^-1 .

methylene chloride) gives the desired compound. IR

b)

(homochiral)

The method of Example 1 is followed, except that in step (d) R—(+)—1— is used instead of benzyl isocyanate.

-phenylpropylisocyanate and then deprotection and workup according to Example 1(e) gives the desired compound. MS (M+H) ⁺

376.1, (2M+H) ⁺ 751.2; IR (KBr) 1780 cm 1 , 1670 cm 1 .

a) 2-naphthylmethyl isocyanate

To a solution of 2-naphthylacetic acid (840 mg, 4.5 mmol) in methylene chloride (15 ml) at room temperature was added a drop of dimethylformamide and oxalyl chloride (1 ml). The resulting mixture was stirred for 30 minutes at room temperature. The solvent was then removed and the residue was dissolved in acetone (15 ml) and cooled to 0 °C. A solution of sodium azide (700 mg, 11 mol) in water (10 ml) was then added. The reaction mixture was then stirred for 30 minutes at 5 °C and then poured into a mixture of ice-cold water (30 ml), ether (40 ml) and hexane (40 ml). The organic phase was separated, washed with brine, dried and concentrated to 5 ml. Chloroform (5 ml) was added to the residue. The resulting solution was added dropwise to chloroform (10 ml) at 80 °C. This mixture was heated at reflux for 1 hour. The solvent was then evaporated to give 680 mg of crude product. Purification by chromatography (silica, methylene chloride) gave 342 mg of the desired product as a white solid. IR (neat product) 2355 cm ^-1 , 2336 cm ^-1 , 2267 cm ^-1 .

b)

(O homochiral)

The method of Example 1 is followed, except that in step (d) 2-naphthylmethyl isocyanate is used instead of benzyl isocyanate, and then deprotection and workup according to Example 1(e) give the desired compound in the form of a flaky powder. MS (M+H) ⁺ 398.1; IR (KBr) 1778 cm ^-1 , 1541 cm ^-1 .

Example 18

About • ·

The procedure of Example 1 is followed, except that in step (d) 4-isocyantomethylbiphenyl is used instead of benzyl isocyanate, and then deprotection and work-up according to Example 1(e) give the desired product. IR (KBr) 1758 cm-1; MS 424.1 (M+H) ⁺ , 422.3 (MH)".

Example 19

(homochiral)

To a mixture of dibenzylamine (2.0 g, 9.8 mmol) and triethylamine (1.2 mL, 85 mmol) in toluene (15 mL) was added a solution of diphosgene (680 μΐ, 5.7 mmol) in toluene. The resulting mixture was stirred for 4 h at room temperature. The mixture was then poured into 2 N aqueous HCl (50 mL) and extracted with ethyl acetate (3 x 50 mL). The organic layers were combined, dried over magnesium sulfate, and concentrated to give 2.60 g of the desired product as a white solid. IR (film) 1731 cm ^-1 .

Oh' • ·

To a solution of the benzyl ester product prepared according to Example 1(c) (116 mg, 0.20 mmol) in methylene chloride (2 mL) were added triethylamine (42 μΐ), 4-dimethylaminopyridine (30 mg) and a solution of the carbamoyl chloride prepared in step (a) (78 mg, 0.30 mmol) in methylene chloride (2 mL). The resulting mixture was stirred for 3 days at room temperature. The reaction was then confirmed to be complete by HPLC. The reaction mixture was then quenched with 1 N sodium hydrogen sulfate (15 mL). The resulting mixture was extracted with ethyl acetate (100 mL). The organic layer was washed with brine (15 mL), dried over magnesium sulfate and concentrated to give the crude product. Purification by flash chromatography (30% ethyl acetate/hexane) gave the desired product (95 mg). MS (M+H) ⁺ 796.1, (MH) 794.4; IR (film) 1785 cm -1 , 1732 cm -1 , 1671 cm ^-1 , 1639 cm ^-1 .

C)

(homochiral)

A mixture of the product prepared in step (b) (95 mg, 0.12 mmol), 1 N HCl (145 μΐ) and the catalyst, 10% palladium on carbon (61 mg) in dioxane (3 ml) was stirred for 2 h at room temperature under a hydrogen atmosphere (hydrogen balloon). Analytical HPLC showed the reaction to be complete at this time. The reaction mixture was then filtered through a pad of Celite ^R and concentrated to give the crude product as a white powder. Purification by reverse phase HPLC using the mobile phase described in Example 1(e) gave the desired product as a white powder (36 mg). MS (M+H) ⁺ • · · · « · · · · *······ · • · « *

I · · ·

438.1, (M-H) 436.3; IR (KBr) 1786 cm“1, 1672 cmi

Example 20

NH»CF ₃ COOH h ₂ n^n

H

£O ₂ H /—\ í?

(homochiral) 'YVJ

N—CO—C(CH ₃ ) ₃

a) (H ₃ C) _:

To a solution of diphosgene (326 μΐ , 20% in toluene) in methylene chloride at 0 °C was added a mixture of tert.butylpiperazinecarboxylate (1.0 g) and triethylamine (753 μΐ) in methylene chloride (5 ml). The resulting mixture was stirred for 90 minutes at 0 OC. The end of the reaction could be confirmed by TLC. The reaction mixture was then quenched with water (20 ml). The organic layer was separated. The aqueous layer was extracted with methylene chloride (2 x 20 ml). The organics were combined and washed with water (10 ml) and brine (2 x 10 ml), dried over magnesium sulfate, filtered and concentrated to give the crude product. Purification by flash chromatography (methylene chloride) gave the desired product as a white solid. IR (KBr) 1680 cm ^-1 , 1747 cm ^-1 .

h)

O

O

O

To a solution of the benzyl ester product prepared according to Example 1(c) (85 mg, 0.15 mmol) in tetrahydrofuran at -78°C was added dropwise sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 180 μΐ). The mixture was stirred for 90 minutes at -78°C. Then a solution of the acid chloride prepared in step (a) (45 mg, 0.18 mmol) in tetrahydrofuran (1 ml) was added. The reaction mixture was stirred for 5 hours at -78°C. Then the reaction was quenched by the addition of 1 N potassium hydrogen sulfate (15 ml). The mixture was then extracted with ethyl acetate (3 x 30 ml). The organic layers were combined and washed with brine (2 x 15 ml), dried over magnesium sulfate, filtered and concentrated to give the crude product. Purification by flash chromatography (30-50% ethyl acetate/hexane) afforded 32 mg of the desired oil as a colorless oil. MS (M+H) ⁺ 785.4, (MH) ^- 783.7; IR (neat) 1786 cm ^-1 , 1732 cm" ¹ , 1681 cm" ¹ ·

C)

O

II

C-ο—C(CH ₃ ) ₃ (homochiral)

Deprotection and purification of the product prepared in step (b) (32 mg) according to the method of Example 19(c) gave 17 mg of the desired product as a flaky powder. MS (M+H) ⁺

427.1, (MH)"425.2; IR (KBr) 1792 cm" ¹ , 1760 cm" ¹ .

• · • · • · ·

Example 21

N— C-(CH ₃ ) ₃ H (homochiral)

a) ff (H ₃ C) ₃ C— o— cn^ o

N—CN—C (CH ₃ ) ₃ / H

To a solution of tert-butyl-1-piperazinecarboxylate (0.5 g, 2.68 mmol) in methylene chloride (10 mL) was added tert-butyl isocyanate (0.28 g, 2.82 mmol) and the mixture was stirred at room temperature. After 2 hours, the reaction mixture was evaporated in vacuo, suspended in water (50 mL) and extracted with ethyl acetate (2 x 50 mL). The organic phase was washed with saturated sodium chloride (1 x 50 mL) and filtered through a glass frit. The filtrate was dried over sodium sulfate and concentrated to give 0.53 g of the desired product as a white solid.

b) ,-< ⁰ /\ II

HN N—C—N—C (CH ₃ ) ₃ \ f H • « • · ·

The product prepared in step (a) (0.475 g, 1.67 mmol) was suspended in methylene chloride (4 ml) and trifluoroacetic acid (4 ml) was added over 1 minute. The resulting mixture was stirred at room temperature. After 1 hour, the mixture was evaporated in vacuo. The residue was dissolved in water, the pH was adjusted to 12-13 and the mixture was extracted with ethyl acetate (2 x 25 ml). The organic phase was washed with saturated sodium chloride (1 x 50 ml), filtered, dried over sodium sulfate and concentrated to give 0.113 g of the desired product as a white solid.

A solution of the benzyl ester product prepared according to Example 1(c) (65 mg, 0.114 mmol) in methylene chloride (1 ml) was cooled to -10 °C and triethylamine (24 μΐ, 0.17 mmol) was added followed by 20% phosgene in toluene (0.15 ml, 0.285 mmol). After 90 minutes at -10 °C, the mixture was evaporated in vacuo. The residue was taken up in methylene chloride (1 ml) and cooled to -10 °C. Triethylamine (24 μΐ, 0.17 mmol) was then added followed by the piperazine product prepared in step (b) (21 mg, 0.114 mmol). The resulting mixture was stirred at -10 °. After 1 hour, the mixture was quenched with 10% potassium dihydrogen phosphate (15 ml) and extracted with ethyl acetate (2 x 20 ml). The organic phase was washed with saturated sodium chloride (1 x 30 ml), dried over sodium sulfate and concentrated to give a light yellow oil. Purification by preparative HPLC (reverse phase, methanol, water, acid • · • · · · · · · · <

trifluoroacetic acid) gave 27 mg of the desired product as a white foam. MS 784.2 (M+H)+; IR (film): 1781.1, 1741.9, 1636.6 cm ^-1 .

A solution of the product prepared in step (c) (25 mg, 0.032 mmol) in 1,4-dioxane (7 mL) was treated with 1 N HCl (40 μΐ, 0.038 mmol) and the catalyst, 10% palladium on carbon (15 mg). Hydrogen gas was then bubbled through the reaction mixture for 2 hours. The reaction mixture was then filtered through a pad of Celite ^R , which was then repeatedly washed with 1,4-dioxane. The combined eluates were lyophilized to give 15 mg of the desired product as a white lyophilisate. MS 426.1 (M+H) ⁺ , 424.3 (MH)-; IR (KBr) 1780 cm-1 .

Example 22 (homochiral)

NH«HC1

O

C(CH ₃ ) ₃ • * θ O

II / \ II pt pifT \ , (H3O3C—O—CN N—C—CH2 C(CH ₃ )3

and)

A solution of tert-butyl-1-piperazinecarboxylate (0.5 g, 2.58 mmol) in methylene chloride (5 mL) was cooled to 0 °C. N,N-diisopropylethylamine (0.42 g, 3.22 mmol) and 4-dimethylaminopyridine (30 mg) were then added followed by tert-butylacetylchloride (0.36 g, 2.68 mmol) over 1 min. The resulting mixture was stirred at 0 °C for 2 h. After 2 h, the mixture was partitioned between water (20 mL) and ethyl acetate (2 x 20 mL). The organic layer was washed with brine (1 x 75 mL), dried over sodium sulfate, and concentrated to give 0.736 g of the desired product as a white solid. MS 285.0 (M+H) ⁺ .

b)

O

N—C- CH ₂ —C(CH ₃ ) ₃

The product from step (a) (0.7 g, 2.46 mmol) was dissolved in methylene chloride (7 ml) and trifluoroacetic acid (4 ml) was added over 1 minute. The resulting mixture was stirred at room temperature. After 1 hour, the mixture was evaporated in vacuo. The residue was dissolved in water, the pH was adjusted to 12-13 and the mixture was extracted with ethyl acetate. The organic phase was washed with brine, dried over sodium sulfate and concentrated to give 0.218 g of the desired product as a pale yellow oil. MS 184.9 (M+H)+.

C) íí

Wed

Cl—CN N—C—CH ₂ —C(CH ₃ ) ₃

A solution of the product prepared in step (b) (109 mg, 0.59 mmol) in methylene chloride (0.5 mL) was added at 0 °C to a mixture of phosgene (0.79 mL of 20% phosgene in toluene, 1.48 mmol) in methylene chloride (2 mL) followed by the addition of triethylamine (82 μΐ, 0.59 mmol). The resulting mixture was stirred for 1 h at 0 °C. The reaction mixture was then partitioned between water (25 mL) and ethyl acetate (2 x 25 mL). The organic phase was washed with 1 N HCl (40 mL), brine (50 mL), dried over sodium sulfate and concentrated to give a brown oil. Purification by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) yielded 70 mg of the desired product.

The product prepared in step (c) (70 mg, 0.122 mmol) was dissolved in tetrahydrofuran (1 mL) and cooled to -78° C. Sodium bis(trimethylsilyl)amide (0.15 mL, 0.146 mmol) was then added over one minute and the mixture was stirred at -78° C. for 1 hour.

A solution of the benzyl ester product prepared according to Example 1(c) (36 mg, 0.146 mmol) in tetrahydrofuran (0.5 ml) was then added and the reaction mixture was stirred at -78 °C. After 2.5 hours, the reaction mixture was quenched with 0.5 N potassium hydrogen sulfate solution (25 ml) and extracted with ethyl acetate (2 x 25 ml). The organic phase was washed with brine (1 x 50 ml), dried over sodium sulfate and concentrated to a yellow oil. Purification by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave 21 mg of the desired product as a colorless oil. MS 783.4 (M+H) ⁺ , 781.3 (MH) ".

E)

NH«HC1

O

Deprotection and workup of the product prepared in step (d) was carried out according to Example 21(d) to give 12 mg of the desired product as a white lyophilisate. MS 425.1 (M+H)+, 423.3 (M-H)-; IR (KBr) 1786, 1736 cm-1.

Example 23

_NH3CF3COOH

A _A CO ₂ H h ₂ nfo ₀ J- _v q -c—(CH ₂ ) ₂ (homochiral)

a) ⁰ O

II /~Λ I?

(H ₃ C) ₃ C— O— C— N— C— (CH ₂ ) ₂

(S°)

To a solution of tert-butyl-1-piperazinecarboxylate (500 mg, 2.68 mmol) in methylene chloride (4 ml) at 0 °C were added N,N-diisopropylethylamine (560 μΐ), 4-dimethylaminopyridine (33 mg) and a solution of 3-phenylpropanoic acid chloride (400 μΐ, 2.69 mmol). The resulting mixture was stirred for 2 hours at 0 °C. The reaction mixture was then quenched with water (20 ml). This mixture was extracted with ethyl acetate (2 x 50 ml). The organics were combined and washed

• · · with brine (2 x 10 mL), dried over magnesium sulfate, and concentrated to give 872 mg of the desired product (crude) as a yellow solid. MS (M+H) ⁺ 319.1.

b) and?

(CH ₂ ) ₂ —C— \_7

NH.CF3COOH

A mixture of the crude product prepared in step (a) (860 mg, 2.68 mmol), trifluoroacetic acid (10 mL) and methylene chloride (10 mL) was stirred for 1 h at room temperature. The reaction was confirmed by TLC. The solvent was then removed and 1 N sodium hydroxide solution (15 mL) was added. The reaction mixture was then extracted with ethyl acetate (100 mL). The combined organic solution was then washed with brine (10 mL), dried over magnesium sulfate, filtered and concentrated to give 238 mg of the desired product as a yellow oil which was used without further purification. IR (film) 1633 cm ^-1 .

A mixture of the product prepared in step (b) (200 mg, 0.92 mmol) and triethylamine (154 μΐ) in methylene chloride (4 mL) was added at 0 °C to a solution of phosgene in toluene (584 μΐ, 20%). The resulting mixture was stirred for 20 min at 0 °C. The completion of the reaction was confirmed by TLC. The solvent was then removed and anhydrous ether was added. The mixture was then filtered and the filtrate was concentrated to give the crude product as a yellow oil. Purification of the crude product by flash chromatography (50% ethyl acetate/hexanes) gave 235 mg of the desired product as a yellow oil. IR (film)

1741 cm-1, 1703 cm-1.

φφφ φ φ · · · • φ φ φφφφ ·

To a solution of the benzyl ester product prepared according to Example 1(c) (100 mg, 0.17 mmol) in tetrahydrofuran (2 ml) was added dropwise at -78 °C a solution of sodium bis(trimethylsilyl)azide (1.0 M in tetrahydrofuran, 210 μΐ, 0.21 mmol). The resulting mixture was stirred for 1 hour at -78 °. Then the product prepared in step (c) (58 mg, 0.21 mmol) in tetrahydrofuran (1 ml) was added. The reaction mixture was then stirred for 2.5 hours at -78 °. Analytical HPLC showed that the starting material had completely reacted. The reaction mixture was then quenched with 1 N potassium hydrogen sulfate (20 ml). The resulting mixture was then extracted with ethyl acetate (2 x 30 ml). The organics were combined, washed with brine (2 x 15 mL), dried (magnesium sulfate), filtered, and concentrated to give the crude product. Purification of the crude product by reverse phase HPLC gave 32 mg of the desired product as a colorless oil. MS (M+H) ⁺ 817.1, (MH) + 815.4.

e) (homochiral)

NH»CF ₃ COOH

H ₂ N^N'

H _to CO ₂ H

O

V • w ·· v * r » »

9 · 9 9 9 ··· ···· · · » · · 9 « 9 9 · ········ · •«99 99

Deprotection and purification of the product prepared in step (d) according to the method of Example 19(c) gave 10 mg of the desired product as a white powder. MS (M+H) ⁺ 459.2, (MH) 457.4; IR (KBr) 1790 cnr i , 1680 cnr i .

Example 24

a) (H ₃ C) ₃ CO—CN í? /—\ sj NC-OCH3

A solution of tert-butyl-1-piperazinecarboxylate (0.5 g, 2.68 mmol) in methylene chloride (5 mL) was cooled to 0 °C. N,N-diisopropylethylamine (0.42 g, 3.22 mmol) and 4-dimethylaminopyridine (30 mg) were then added followed by methyl chloroformate (0.25 g, 2.69 mmol) over 1 min. The mixture was stirred at 0 °C for 1 h. The mixture was then partitioned between water (20 mL) and ethyl acetate (2 x 20 mL). The organic phase was washed with brine (1 x 75 mL), dried over sodium sulfate and concentrated to give the desired product as a cream solid (0.636 g).

__ _to O

b) f ^{11 J} HN N—C— OCH ₃

The product prepared in step (a) (0.3 g, 1.23 mmol) was dissolved in methylene chloride (3 ml) and cooled to 0 °C. Trifluoroacetic acid (3 ml) was then added, the mixture was warmed to room temperature and stirred for 1 hour. The mixture was then evaporated in vacuo. The residue was dissolved in water, the pH was adjusted to 12-13 with 6 N sodium hydroxide, and extracted with ethyl acetate. The organic phase was washed with brine, dried over sodium sulfate, and concentrated to give 91 mg of the desired product free of the amine. IR (film) 1696.2 cm ^-1 ; MS 144.9 (M+H) ⁺ .

C)

Cl—C-

N— C-OCH3

II

A mixture of the product prepared in step (b) (89 mg, 0.617 mmol) and triethylamine (86 μΐ, 0.59 mmol) in methylene chloride (2 mL) was added to a mixture of phosgene (0.82 mL of a 20% solution of phosgene in toluene, 1.54 mmol). This mixture was stirred for 1 h at 0 °C. The reaction mixture was then partitioned between water (25 mL) and ethyl acetate (2 x 25 mL). The organic phase was washed with 1 N HCl (40 mL), brine (50 mL), dried over sodium sulfate and concentrated to give 108 mg of the desired product as a brown oil. IR (film) 1738.8, 1704.7 cnr».

O

The benzyl ester product prepared according to Example 1(c) (111 mg, 0.194 mmol) was dissolved in tetrahydrofuran (2 mL) and cooled to -78 °C. Then bis(trimethylsilyl)amide (0.23 mL, 0.234 mmol) was added over 1 min and the mixture was stirred at -78 °C for 1 h. Then a solution of the product prepared in step (c) (48 mg, 0.234 mmol) in tetrahydrofuran was added and the reaction mixture was stirred at -78 °C. After one hour, the mixture was quenched with 0.5 N potassium hydrogen sulfate (25 mL) and extracted with ethyl acetate (2 x 25 mL). The organic phase was then washed with brine (1 x 50 mL), dried over sodium sulfate and concentrated to give a pale yellow oil. Purification by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave 25 mg of the desired product as a colorless oil/foam. MS 743.1 (M+H) ⁺ , 741.4 (MH) - ; IE (film) 1786.6 cm -1

E)

O

Deprotection of the product prepared in step (d) (22 mg, 0.027 mmol) and work-up according to the method of Example 21(d) afforded 12 mg of the desired product as a white lyophilisate.

MS 385.1 (M+H) ⁺ , 383.2 (M-H)

IR (film) 1786 cm.

Example 25 (homochiral)

and)

* · • ·

To a mixture of phosgene (5.5 mL of a 20% phosgene in toluene solution, 11.01 mmol) in methylene chloride (5 mL) at 0 °C was added 3-phenyl-1-propanol (1.09 g, 7.34 mmol). The mixture was stirred at 0 °C for 5 h. The reaction mixture was then evaporated in vacuo to give a colorless oil. Purification by flash chromatography (silica gel, 0-5% ethyl acetate/hexane) gave 1.33 g of the desired product.

b)

O( _H3C ) ₃ —-co— c-

A solution of tert-butyl-1-piperazinecarboxylate (0.2, 1.07 mmol) in methylene chloride (3 mL) was cooled to 0 °C. N,N-diisopropylethylamine (0.25 g, 1.93 mmol) was then added and

4-dimethylaminopyridine (5-7 crystals) and then a solution of the product prepared in step (a) (0.2 g, 1.07 mmol) was added over 1 minute. This mixture was stirred for 1 hour at 0°C. The mixture was then partitioned between water (20 mL) and ethyl acetate (2 x 20 mL). The organic layer was washed with brine (1 x 75 mL), dried over sodium sulfate and concentrated to give 0.35 g of the desired product as a yellow oil.

C)

The product prepared in step (b) (0.35 g, 1.03 mmol) was dissolved in methylene chloride (4 ml) and cooled to 0 °C. Trifluoroacetic acid (4 ml) was then added over 1 minute, the mixture was warmed to room temperature and stirred for 1 hour. The mixture was then evaporated in vacuo. The residue was dissolved in water, the pH was adjusted to 12-13 with 6 N sodium hydroxide, and the mixture was extracted ·« 90 9 ··

0» * · V 909 ··♦· · » · · · 9 • » ··· ···»·*· *

009999 ·· · ·· with ethyl acetate. The organic phase was washed with brine, filtered over sodium sulfate and concentrated to give 0.23 g of the desired product as a white solid. MS 248.9 (M+H) ⁺ .

d)

Cl—

II c—

O

II co— (CH ₂ ) ₃

To a mixture of phosgene (0.53 mL of a 20% solution of phosgene in toluene, 1.01 mmol) was added a solution of the product prepared in step (c) (100 mg, 0.403 mmol) in methylene chloride (1 mL) followed by the addition of triethylamine (60 μΐ, 0.403 mmol). The mixture was stirred for 1 h at 0 °C. The reaction mixture was then partitioned between water (25 mL) and ethyl acetate (2 x 25 mL). The organic phase was then washed with 1 N HCl (40 mL), brine (50 mL), dried over sodium sulfate and concentrated to give 115 mg of the desired product.

O

A solution of the benzyl ester product prepared in Example 1(c) (43 mg, 0.075 mmol) in methylene chloride (1 ml) was cooled to 0°C and triethylamine (11 mg, 0.113 mmol) and 4-dimethylaminopyridine (6-8 crystals) were added. A solution of the product prepared in step (d) (58 mg) in methylene chloride (0.5 ml) was then added and the mixture was stirred for 45 minutes at 0°C and then for 3 hours at room temperature. The resulting mixture was evaporated in vacuo. The residue was purified by flash column chromatography ·· ·· • 9 9

999

9*

4449 ii ·· · ·· • ft · ·

9*4 9 9 9

9 9999949 9 9

4 9 9 9 » ·· · ·« · 9 · (silica gel, 30% ethyl acetate/hexanes) gave 40 mg of the desired product as a light yellow oil. MS 847.1 (M+H) ⁺ , 845.4 (MH) - ; IR (film) 1784 cm -*.

(homochiral)

O

Deprotection of the product prepared in step (e) (40 mg, 0.027 mmol) and treatment according to Example 21(d) gave 21 mg of the desired product as a white lyophilisate. MS 489.1 (M+H)*, 487.4 (MH)-; IR (KBr) 1784, 1667 cm ^-1 .

Example 26

(homochiral)

a) (H ₃ C) ₃ —C—O—C-

To a mixture of phosgene (3.6 mL of a 20% phosgene in toluene solution, 6.71 mmol) in methylene chloride (2 mL) at 0 °C was added a solution of tert-butyl-1-piperazinecarboxylate (0.5 g, 2.8 mmol) in methylene chloride (1 mL). Triethylamine (0.27 g, 6.71 mmol) was then added and the mixture was stirred for 1 h at 0 °C. The mixture was then partitioned between water (30 mL) and ethyl acetate (2 x 30 mL). The organic layer was washed with brine (1 x 60 mL), dried over sodium sulfate, and concentrated to give the crude product. Purification of the crude product by flash chromatography (silica gel, 40% ethyl acetate/hexane) gave 0.515 of the desired product as a white solid. IR (film) 1737.6, 1697.0 cm” ¹ .

( _CH3 ) ₃

A solution of the benzyl ester product prepared according to Example 1(c) (47 mg, 0.082 mmol) in methylene chloride (1 ml) was cooled to 0 °C and then triethylamine (12 mg, 0.123 mmol) and 4-dimethylaminopyridine (6-8 crystals) were added. A solution of the product prepared in step (a) (51 mg) in methylene chloride (1 ml) was then added and the mixture was stirred for 40 minutes at 0 °C and then for 4-5 hours at room temperature. The mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 48 mg of the desired product as a colorless oil. MS 785.1 (M+H) ⁺ , 783.4 (MH) ^- .

C)

O

O

CO2-CH2

n_^nh«cf ₃ cooh

The product prepared in step (b) (40 mg, 0.051 mmol) was dissolved in methylene chloride (1 mL) and cooled to 0 °C. Trifluoroacetic acid was then added over 1 min and the mixture was warmed to room temperature and stirred for 1 h. The mixture was then evaporated in vacuo to give the desired product as a yellow oil which was used in the next step without further purification. MS 685.1 (M+H) ⁺ , 683.3 (MH) ^- .

d)

NH«HC1

_H2N ^NH

> _CO2H

I (homochiral) ^{/ 111'HC1O}

Deprotection and treatment of the product prepared in step (c) (49 mg, 0.061 mmol) according to the method of Example 21(d) gave 15 mg of the desired product as a white lyophilisate. MS 327.0 (M+H)*, 325.0 (MH) ^- ; IR (KBr) 1786, 1653 cm - .

Example 27

NH«HC1

(homochiral) (h ₃ o ₃ —co—cn^_n-so ₂ —ςθ/

A solution of tert-butyl-1-piperazinecarboxylate (0.2 g, 1.07 mmol) in methylene chloride (2 mL) was cooled to 0 °C. N,N-diisopropylethylamine (0.167 g, 1.28 mmol) was then added and

I ί » · · • · · · • · · · · ····· · · ·

4-dimethylaminopyridine (30 mg) was added followed by benzenesulfonyl chloride (0.19 g, 1.07 mmol) over 1 minute. The mixture was stirred at 0 °C for 2 hours. After 2 hours, water (20 mL) was added and the mixture was extracted with ethyl acetate (2 x 20 mL). The organic layer was then washed with brine (1 x 75 mL), dried over sodium sulfate and concentrated to give 0.35 g of the desired product as a white solid.

The product prepared in step (a) (0.35 g, 1.07 mmol) was dissolved in methylene chloride (3 ml) and cooled to 0 °C. Trifluoroacetic acid was then added over 1 min, the mixture was warmed to room temperature and stirred for 1 h. The mixture was then evaporated in vacuo. The residue was dissolved in water, the pH was adjusted to 12-13 with 6 N sodium hydroxide and the mixture was extracted with ethyl acetate. The organic phase was washed with brine, dried over sodium sulfate and concentrated to give 0.208 g of the desired product as a pale yellow oil. MS 226.8 (M+H) ⁺ .

A mixture of the product prepared in step (h) (100 mg, 0.442 mmol) and triethylamine (62 μΐ, 0.442 mmol) in methylene chloride (1 mL) was added to a mixture of phosgene (0.59 mL of a 20% solution of phosgene in toluene, 1.1 mmol). The mixture was stirred for 1 h at 0 °C. The mixture was then evaporated in vacuo. The residue was suspended in ether and the mixture was filtered. The eluates were concentrated to give 100 mg of the desired product as a cream-colored solid.

d)

O

A solution of the benzyl ester product prepared according to Example 1(c) (64 mg, 0.112 mmol) in methylene chloride (1 mL) was cooled to 0 °C and then triethylamine (17 mg, 0.168 mmol) and 4-dimethylaminopyridine (6-8 crystals) were added. The product prepared in step (c) (58 mg, 0.168 mmol) was then added and the mixture was stirred for 45 min at 0 °C and then for 3 h at room temperature. The mixture was then evaporated in vacuo to give the crude product. Purification of the crude product by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave 28 mg of the desired product as a colorless oil. MS 825.1 (M+H) ⁺ .

e) (homochiral)

NH»HCl

Deprotection and treatment of the product prepared in step (d) (25 mg, 0.03 mmol) according to the method of Example 21(d) gave 12 mg of the desired product as a white lyophilisate. MS 467.0 (M+H) ⁺ , 465.3 (MH) - ; IR (film) 1787.25, 1662.13 cm 1 .

Example 28

NH4HC1 11

H ₂ NNH £0 ₂ H

O li • · · · · · «··· · · · · • · · * · «···« « · · · » * • · · « «· ·· · (homochiral)

NN— C- O- CH ₂ - CH (CH ₃ ) ₂ <_/

and)

O

II (H ₃ C) ₃ C—O—CNN CO— CH ₂ —CH(CH ₃ ) ₂ \_/

To a solution of tert-butyl piperazine carboxylate (500 mg, 2.68 mmol) in methylene chloride (2 mL) at 0 °C were added diisopropylethylamine (30 mg), 4-dimethylaminopyridine (30 mg) and a solution of isobutyl chloroformate (366 μΐ, 268 mmol) in methylene chloride (2 mL). The mixture was stirred for 1 h at 0 °C. The reaction mixture was then quenched with water (20 mL). The mixture was extracted with ethyl acetate (2 x 50 mL). The organics were combined and washed with brine (2 x 10 mL), dried over magnesium sulfate and concentrated to give 819 mg of the desired product as a yellow solid; IR (film) 1701 cm ^-1 , 1688 cm ^-1 .

b)

O

CO-CH ₂ -CH(CH ₃ )

A mixture of the crude product prepared in step (a) (800 mg, 2.79 mmol), trifluoroacetic acid (10 mL) and methylene chloride (10 mL) was stirred for 1 h at room temperature. After this time, the reaction was shown to be complete by TLC. The solvent was then removed and 1 N sodium hydroxide solution (15 mL) was added. The mixture was then extracted with ethyl acetate (100 mL). The combined organics were washed with brine (10 mL), dried over magnesium sulfate and concentrated to give 511 mg of the desired product as a yellow oil, which was used without further purification. IR (film) 1692 cm- ¹ .

C)

O

N—C-O-CH ₂ -CH (CH ₃ ) ₂

O

II

To a solution of phosgene in toluene (1.6 ml, 20%) was added a mixture of the product prepared in step (b) (477 mg, 2.56 mmol) and triethylamine (432 μΐ) in methylene chloride (5 ml) at 0 °C. The resulting mixture was stirred for 2 hours at 0 °C. After this time, the reaction was complete by TLC. The solvent was then removed and anhydrous ether (50 ml) was added. The mixture was then filtered and the filtrate was concentrated to give the crude product (481 mg) as an orange oil. Purification of the crude product gave 453 mg of the desired product as a yellow oil. IR (film) 1741 cmi, 1703 cmi.

d)

II

To a solution of the benzyl ester product prepared according to Example 1(c) (100 mg, 0.17 mmol) in methylene chloride (1 ml) were added triethylamine (36 μΐ), 4-dimethylaminopyridine (30 mg) and a solution of the product prepared in step (c) (65 mg, 0.24) in methylene chloride (1 ml). The resulting mixture was stirred for 4 hours at room temperature. After this time, the reaction was confirmed to be complete by analytical HPLC. The reaction mixture was then quenched with 1 N potassium hydrogen sulfate (15 ml). The mixture was then extracted with ethyl acetate (100 ml). The organic layer was washed with brine (15 ml), dried over magnesium sulfate and concentrated to give the crude product as a yellow oil. Purification

100 « · flash chromatography (50% ethyl acetate/hexane) afforded 81 mg of the desired product. MS 785.2 (M+H) ⁺ , 783.4 (MH) ^- .

E)

(homochiral

O ___{>-o-ch ₂ -ch(ch ₃ )2

and)

Deprotection and treatment of the product prepared in step (d) (80 mg, 0.10 mmol) according to the method of Example 21(d) gave 41 mg of the desired product as a white solid. MS (M+H) ⁺ 427.1, (MH) - 425.3; IR (KBr) 1786 cm, 1653 cm -1 .

Example 29

The desired product was prepared as an orange oil by the method of Example 25(a) to (d) except that phenethyl alcohol was used in place of 3-phenyl-1-propanol in step (a).

b)

O

101 ···» · «·· · · • · ··· ·····« ·

A solution of the benzyl ester product prepared according to Example 1(c) (69 mg, 0.121 mmol) in methylene chloride (1 ml) was cooled to 0 °C and triethylamine (18 mg, 0.181 mmol) and 4-dimethylaminopyridine (6-8 crystals) were added. A solution of the product prepared in step (a) (54 mg) in methylene chloride (1 ml) was then added and the mixture was stirred at 0 °C for 45 min and then at room temperature for 2.5 h. The mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 90 mg of the desired product as a colorless oil. MS 833.1 (M+H)+, 831.4 (M-H)-; IR (film) 1786, 1737.7 ctiri.

C)

NH»HC1 11

H ₂ NN'

H (homochiral)

K >co ₂ h U t—\ ' Γ O—(CH ₂ ) ₂ —(O)

J—·γ·υ

Deprotection and treatment of the product prepared in step (b) (85 mg, 0.108 mmol) according to the method of Example 21(d) gave 34 mg of the desired product as a white lyophilisate. MS 475.1 (M+H) ⁺ , 473.4 (MH) - ; IR (film) 1783 cnr i , 1665 cnr i .

Example 30 (homochiral)

NH»CF ₃ COOH

O

102

a) Cyclohexyl chloroformate

To a solution of phosgene in toluene (5.3 ml, 20%) was added a mixture of cyclohexanol (500 mg, 5.0 mmol) and triethylamine (836 μΐ) in methylene chloride (4 ml) at 0 °C. The resulting mixture was stirred for 2.5 hours at 0 °C. After this time, the end of the reaction could be demonstrated by TLC. The solvent was then removed and anhydrous ether (50 ml) was added. The mixture was then filtered and the filtrate was concentrated to give 726 mg of the desired product as a colorless oil, which was used without further purification. IR (film) 1776 cm“ ¹ .

About _{_} _ About

b) II /\ II (CH ₃ ) 3 — c— o— CN— c- O

To a solution of tert-butyl-1-piperazinecarboxylate (500 mg, 2.68 mmol) in methylene chloride (3 mL) at 0 °C were added diisopropylethylamine (560 μΐ), 4-dimethylaminopyridine (30 mg) and a solution of cyclohexyl chloroformate (473 mg, 2.68 mmol) in methylene chloride (2 mL). The resulting mixture was stirred at 0 °C and then allowed to warm to room temperature over 4 h. The reaction was then quenched by addition of water (15 mL). The reaction mixture was then extracted with ethyl acetate (100 mL). The organic layer was washed with brine (2 x 10 mL), dried over magnesium sulfate and concentrated to give 832 mg of the desired product as a white solid. IR (film) 1692 cm ^-1 .

^C) r~\ í?

HN N-C—

A mixture of the crude product prepared in step (b) (832 mg, 2.68 mmol), trifluoroacetic acid (2 mL) and methylene chloride (2 mL) was stirred at room temperature for 3 hours.

103

room temperature. After this time, the reaction was complete by TLC. The solvent was then removed and 1N sodium hydroxide solution (15 mL) was added. The mixture was then extracted with ethyl acetate (100 mL). The combined organic solution was then washed with brine (10 mL), dried over magnesium sulfate, filtered, and concentrated to give 535 mg of the desired product as a pale yellow oil, which was used without further purification.

^d) í? i-< ⁰

II /\ II Cl—C N— C- O

To a solution of phosgene in toluene (2.1 ml, 20%) was added at 0 °C a mixture of the product prepared in step (c) (459 mg) and triethylamine (448 μΐ) in methylene chloride (2 ml). The resulting mixture was stirred for 1 hour at 0 °C. After this time, the reaction was confirmed to be complete by TLC. The solvent was then removed and anhydrous ether (50 ml) was added. The mixture was then filtered and the filtrate was concentrated to give the desired product (626 mg) as an orange oil. Purification of the crude product gave 626 mg of the desired product as a yellow solid. IR (film) 1740 cm ^-1 , 1697 cm“ ¹ .

About About

To a solution of the benzyl ester product prepared according to Example 1(c) (113 mg, 0.14 mmol) in methylene chloride (1 mL) was added

104

triethylamine (34 μΐ), 4-dimethylaminopyridine (20 mg) and a solution of the product prepared in step (d) (65 mg, 0.24 mmol) in methylene chloride (1 ml) are added. The mixture is then stirred for 2 hours at room temperature. After this time, the end of the reaction can be demonstrated by analytical HPLC. The reaction mixture is quenched with potassium hydrogen sulfate (15 ml). The mixture is then extracted with ethyl acetate (100 ml). The organic layer is then washed with brine (15 ml), dried (magnesium sulfate) and concentrated to give the crude product as a colorless oil. Purification of the crude product by flash chromatography (50% ethyl acetate/hexane) gives 113 mg of the desired product. IR (film) 1786 cm ^-1 , 1734 cm ^-1 , 1683 cm ^-1 , 1639 cm ^-1 .

f) (homochiral)

NH»CF ₃ COOH

HER

H ₂ NN ² H

, _CO2H

Deprotection and purification of the product prepared in step (e) (113 mg, 0.14 mmol) according to the method of Example 19(c) gave 33 mg of the desired product as a white solid. MS (M+H)* 453.3, (M-H)" 451.5; IR (KBr) 1790 cm 1, 1674 cm 1 .

Example 31 (homochiral)

NH-HC1 at

H ₂ NN £O ₂ H

.C—C(CH ₃ ) ₃

105 • ·

and)

O

II cl—c--Ν N—CC(CH ₃ ) ₃

The desired product was prepared as a mass by the method of Example 22(a) to (c) using tert-butyl carbonyl chloride instead of tert-butyl acetyl chloride in step (a). IR (film) 1733.2, light brown solid except that 1616.4 cm-1 was used.

To a solution of the benzyl ester product prepared according to Example 1(c) (90 mg, 0.157 mmol) in methylene chloride (1 mL) was added triethylamine (24 mg, 0.236 mmol) and

4-dimethylaminopyridine (8-10 crystals). A solution of the product prepared in step (a) (59 mg, 0.236 mmol) in methylene chloride (1 ml) was then added and the mixture was stirred for 30 minutes at 0 °C and then for 6 hours at room temperature. The mixture was then evaporated in vacuo. Purification by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 89 mg of colorless product. MS

769.4 (M+H)+, 767.6 (M-H)-; IR (Film) 1785.4, 1733.4, 1679.4,

1635.9 cm ^-1 .

c) (homochiral) h ₂ n

NH«HC1

£O ₂ H o

II

106 ··· · · · * · ···· * ··· · ·

1 9 9 9 ········ ·

Deprotection and treatment of the product prepared in step (b) (87 mg, 0.11 mmol) according to the method of Example 21(d) gave 10 mg of the desired product as a white lyophilized solid. MS 411.2 (M+H) ⁺ , 409.5 (MH) - ; IR (KBr) 1788.0, 1742.0 cm -1 .

Example 32

The product was prepared according to Example 21 as follows:

can also be prepared

and)

O li (H3O3C-O-C-

g—c(ch ₃ )

To a solution of tert-butyl-1-piperazinecarboxylate (9.17 g, 49.2 mmol) in methylene chloride (40 ml) at 0 °C under nitrogen, a solution of tert-butyl isocyanate (10.28 g, 103 mmol) in methylene chloride (20 ml) was added over 2 minutes. The reaction mixture was stirred for 2 hours at room temperature and then poured into hexane (60 ml). The precipitate formed was filtered off and washed with hexane/methylene chloride (2:1 (2 x 50 ml). The combined eluted fractions were concentrated to a volume of about 20 ml, the precipitate formed was filtered off and washed as described above and combined with the solid fraction obtained above. The solid fraction was then dried in vacuo to give 14.1 g of the desired product. MS 286.2 (M+H) ⁺ .

b) /~\

O

II H

C— N— C(CH ₃ )

107 » · * • · • · · ·

To a solution of the product prepared in step (a) (14.1 g, 49 mmol) in methylene chloride (25 mL) at 0 °C under nitrogen was added dropwise over 5 min. Trifluoroacetic acid (25 mL) was then stirred at 0 °C for 20 min and then at room temperature for 2 h. The reaction mixture was then transferred to a beaker (100 mL) and diluted with ethyl acetate (200 mL) and water (200 mL). Sodium hydroxide solution (25% aqueous) was then added dropwise to the biphasic mixture with vigorous stirring until the pH of the aqueous phase reached about 12. The organic phase was then separated with further addition of ethyl acetate (200 mL). The combined organics were then dried over sodium sulfate, filtered and concentrated to give 11 mg of the desired product as a white solid.

C)

NO

II H c—NC(CH ₃ ) ₃

To a solution of phosgene (20% in toluene, 70 ml, 132 mmol) at 0 °C under nitrogen, a solution of the product prepared in step (b) (ca. 11 g) in methylene chloride/acetonitrile (1:1, 40 ml) was added dropwise over 10 minutes. Triethylamine (30 ml) was then added dropwise over 5 minutes. The reaction mixture was then stirred for 2 hours at room temperature. The reaction mixture was then transferred to a separatory funnel, diluted with ethyl acetate (150 ml) and washed with 2 N HCl (2 x 150 ml). The organics were dried over sodium sulfate, filtered and concentrated to give a yellow oil. Purification by flash chromatography (silica gel,

0-50% ethyl acetate in hexane) gave 7.8 g of the desired product.

108 • · • · · « · ·

To a solution of the benzyl ester product prepared in Example 1(c) (6.94 g, 12.13 mmol) in methylene chloride (50 mL) at room temperature under nitrogen was added the carbamoyl chloride product prepared in step (c) (4.5 g, 18.2 mmol), triethylamine (2.6 mL, 18.2 mmol) and dimethylaminopyridine (225 mg). The reaction mixture was stirred at room temperature for 4 hours and then another portion of the acid chloride product prepared in step (c) (1 g, 4 mmol) and another portion of triethylamine (1 mL, 7 mmol) were added. The reaction mixture was then stirred for an additional 3 hours. The reaction mixture was then diluted with hexane (5 mL) and the crude reaction mixture was loaded onto a silica column (wet with hexane) and purified by flash chromatography (0 to 60% ethyl acetate in hexane) to give 7.46 g of the desired product. MS 784.4 (M+H) ⁺ , 782.2 (MH) ^- .

E)

To a solution of the product prepared in step (d) (7.46 g, 9.57 mmol) in dioxane (125 mL) at room temperature under nitrogen was added water (50 mL), concentrated HCl (0.8 mL, 9.6 mmol) and 10% palladium on carbon catalyst (7.5 g, 50% water content). Hydrogen was then bubbled through the solution for 30 min and

109

I ·· ♦ ft • ftftft

then the reaction mixture is stirred under a hydrogen atmosphere (1 atmosphere) for hours. The reaction mixture is then filtered through a pad of Celite ^R , which is then washed with water (about 100 ml) until the product is detected in the eluate. The solution is then frozen and lyophilized to obtain

4.5 g of white solid. Purification by HPLC (reverse phase, methanol, water, trifluoroacetic acid), followed by lyophilization, filtration through polyvinylpyridine using water as the mobile phase, and final lyophilization gave 3.3 g of the desired product as a bulky white solid. MS 426.2 (M+H)♦, 424.4 (M-H)-; IR (KBr) 1777 cm-1.

Analysis: calculated for Ci8H31N7O5.1.56 H2O:

C, 47.66; H, 7.58; N, 21.62; O, 23.14; found:

C, 47.58; H, 7.37; N, 21.41.

Example 33

_NH3CF3COOH

O

and)

II / \ IÍ Ζ-Λ (H ₃ C) ₃ C—O—C—1S^ /N c— Cr ₂ \O/

The title compound was prepared as a yellow solid by the method of Example 23(a) using phenylacetyl chloride instead of 3-phenylpropanoic acid chloride.

110

A mixture of the product prepared in step (a) (2.68 mmol), trifluoroacetic acid (10 ml) and methylene chloride (10 ml) was stirred for 90 minutes at room temperature. After this time, the reaction was confirmed to be complete by TLC. The solvent was then removed and 1 N sodium hydroxide solution (10 ml) was added. The mixture was then extracted with ethyl acetate (100 ml). The organic solution was then washed with brine (20 ml), dried (magnesium sulfate), filtered and concentrated to give 536 mg of the desired product as a colorless oil which was used without further purification. IR (film) 1630 cm ^-1 .

C)

To a solution of phosgene (351 μΐ, 20% in toluene) in methylene chloride (1 ml) at 0 °C was added a mixture of the product prepared in step (b) (113 mg, 0.55 mmol) and triethylamine (92 μΐ) in methylene chloride (1 ml). The resulting mixture was stirred for 2 hours at 0 °C and then worked up as in Example 23(c) to give 74 mg of the desired product as a yellow solid. IR (film) 1735 cm“ ¹ , 1645 cm“ ¹ .

d)

O

111

» «« • · • · · ·

To a solution of the benzyl ester product prepared according to Example 1(c) (113 mg, 0.20 mmol) in methylene chloride (1 mL) were added triethylamine (34 μΐ), 4-dimethylaminopyridine (30 mg) and a solution of the product prepared in step (c) (74 mg, 0.28 mmol) in methylene chloride (2 mL). This mixture was stirred for 2 hours at room temperature. The reaction was then determined to be complete by analytical HPLC. The reaction mixture was then quenched with 1 N potassium sulfate (10 mL). The mixture was then extracted with ethyl acetate (100 mL). The organic layer was washed with brine (15 mL), dried (magnesium sulfate) and concentrated to give the crude product as a colorless oil. Purification by flash chromatography (30-50% ethyl acetate/hexane) yielded 73 mg of the desired product. MS (M+H) ⁺ 803.4, (MH) ^- 801.5; IR (film) 1785 cm-i, 1733 cm'i, 1733 cmi, 1677 cm'i, 1640 cm-i.

(homochiral)

E)

About II

Deprotection and workup of the product prepared in step (d) (67 mg, 0.83 mmol) according to the method of Example 19(c) gave 5 mg of the desired product as a white solid. MS (M+H) ⁺ 445.2, (MH) - 443.4; IR (film) 1782 cm -1 , 1677 cm -1 .

Example 34 (homochiral)

NH4HC1 11

H ₂ NN, every ₂ hours

—o-ch ₂ oo

112 • ·

a) ιι / \ ii

Cl C— N\ N C— O—

The desired compound was prepared as a yellow oil by the method of Example 30(a) to (d), except that cyclohexylmethanol was used in place of cyclohexanol in step (a). IR (film) 1702 cm ^-1 .

• from 1743 cm ^-1 is used,

b)

O

The product prepared in step (a) was reacted with the benzyl ester product prepared in Example 1(c) according to the method of Example 30(e) to give the desired product as a colorless oil. IR (film) 1786 cm ^-1 , 1732 cm ^-1 , 1680 cm ^-1 , 1639 cm ^-1 .

C)

NH»HC1 (homochiral)

O

Deprotection and condensation of the product prepared in step (b) according to the method of Example 21(d) gave the desired product as a white solid. MS (M+H) ⁺ 467.3, (MH) ^- 465.5; IR (KBr) 1778 cm ^-1 , 1541 cm ^-1 .

113 • · ·· *· · • « · · · ·<· ·«··· · ·

Example 35

O (homochiral)

and)

r—O—C ₂ H ₅

O'

To a stirred solution of the azetidinone product prepared according to Example 1(h) (40 mg, 0.83 mmol) and iodoethane (27 μΐ, 0.332 mmol) in dimethylformamide (200 μΐ) at room temperature was added cesium carbonate (14 mg, 0.042 mmol). After three hours, the reaction mixture was partitioned between ethyl acetate and water containing a small amount of sodium thiosulfate. The organic phase was separated, washed with saturated sodium chloride, dried over magnesium sulfate, and concentrated. The residue was purified by silica gel chromatography to give 33 mg of the desired product.

b)

O

II

O'

O

114 • ·

The product prepared in step (a) (86 mg, 0.168 mmol) and piperazinylcarbamoyl chloride of the formula ° /—\ Π cl-C— MC- N- C (CH ₃ ) ₃ (56 mg, 0.227 mmol) [prepared according to the method of Example 32(c)] were dissolved in methylene chloride (1.80 mL) and tetrahydrofuran (0.20 mL). Triethylamine (35 μΐ, 0.252 mmol) was then added followed by 4-dimethylaminopyridine (4.0 mg, 0.034 mmol). After 48 hours, the reaction mixture was concentrated and the crude product was purified by silica gel chromatography to give 71 mg of the desired product.

c) (homochiral)

NH«HC1

X h ₂ n^n

C— O-~ ^c 2 ^h 5 n _n -cn—C(CH ₃ ) ₃ ii ^H o

Deprotection and treatment of the product prepared in step (b) according to the method of Example 21(d) gave the desired product in the form of a lyophilisate. IR (KBr) 1788 cm ^-1

Example 36 nh*cf ₃ cooh

H ₂ N^ _>CO ₂ H

H ¹ (homochiral)

II CO-CHoCH ₂ -CH(CH ₃ ) ₂

115 • · about

N^/NH

and)

O

II (H3O3—c-o-c—

To a solution of homopiperazine (400 mg, 4.99 mmol) in tetrahydrofuran (40 ml) was added dropwise a solution of di-tert.butyldicarbonate (1.09, 4.99 mmol) and triethylamine (700 μΐ, 4.99 mmol) in tetrahydrofuran (15 ml) over 20 minutes. The reaction mixture was then stirred for 2 hours at room temperature. The mixture was then filtered and the filtrate was concentrated to give the crude product as a colorless oil. Purification by flash chromatography (5% 2 N ammonia in methanol/methylene chloride) gave 410 mg of the desired product as a colorless oil. IR (film) 1691 cm ^-1 .

b) ^{0 0}

II / \ II (H ₃ C) 3 C- O- C— N^ NH— c- O-CH ₂ —CH (CH ₃ ) ₂

To a solution of the product prepared in step (a) (374 mg, 1.87 mmol) in methylene chloride (2 mL) were added diisopropylethylamine (380 μΐ), 4-dimethylaminopyridine (35 mg) and a solution of isobutyl chloroformate (242 μΐ, 1.87 mmol) in methylene chloride (2 mL). The mixture was stirred at 0 °C and allowed to warm to room temperature overnight. The reaction mixture was then quenched with water (15 mL) and worked up as in Example 28(a) to give 508 mg of the desired product. IR (film) 1696 cm“ ¹ .

C)

About

NC-o-CH ₂ —CH (CH ₃ ) 2 • ·

116 * · · ··· ·· ···· · ·«· · · · • · · · · · ······· · • · · ··· ··· ····· ·· ···

The product prepared in step (b) (499 mg, 1.66 mmol) was treated with trifluoroacetic acid (2 mL) according to the method of Example 28(b) to give 324 mg of the desired product as a colorless oil which was used without further purification. IR (film) 1695 cm- ¹ .

d)

O

II ci—co \_y ^N

II

CO-CH ₂ —CH(CH ₃ ) ₂

The product prepared in step (c) (303 mg, 1.51 mmol) was treated at 0 °C with phosgene in toluene (1.2 ml, 20%) according to the method of Example 28(c) to give 298 mg of the desired product as a colorless oil. IR (film) 1738 cm-1, 1697 cm ^-1 .

To a solution of the benzyl ester product prepared according to Example 1(c) (113 mg, 0.20 mmol) in methylene chloride (1 mL) were added triethylamine (34 μΐ), 4-dimethylaminopyridine (10 mg) and a solution of the product prepared in step (d) (62 mg, 0.24 mmol) in methylene chloride (2 mL). The mixture was stirred overnight at room temperature. After this time, the end of the reaction could be indicated by analytical HPLC. The reaction mixture was then quenched with 1 N potassium hydrogen sulfate (15 mL) and workup according to Example 28(d) and subsequent purification afforded 92 mg of the desired product as a colorless oil. MS (M+H)+ 799.4, (M-H) 797.6.

117

F)

NH'CF ₃ COOH h ₂ n^n

H

_to what ₂ H

• · · · · · (homochiral)

c-0- ch ₂ —CH(CH ₃ ) 2

Deprotection and treatment of the product prepared in step (e) (81 mg, 0.10 mmol) according to the method of Example 19(c) gave 5 mg of the desired product as a colorless glass.

MS (M+H) ⁺ 441.3, (MH) - 439.4; IR (film) 1784 cm ¹ , 1665 cnr i.

Example 37

and)

(homochiral)

To a solution of the benzyl ester product prepared according to Example 1(c) (80 mg, 0.14 mmol) in tetrahydrofuran (2 mL) at -78 °C was added dropwise sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 154 μΐ, 0.15 mmol). The resulting mixture was stirred for 40 min at -78 °C. Then a solution of

of 2-thiophenecarbonyl chloride (34 μΐ, 0.30 mmol) in

118

tetrahydrofuran. The reaction mixture was then stirred at -78 °C for a further 6 hours and then left in a freezer (-50 °C) overnight. Analytical HPLC indicated that the reaction was complete. The reaction mixture was then quenched with saturated aqueous ammonium chloride (5 mL). The mixture was then extracted with ethyl acetate (3 x 15 mL). The organics were combined, washed with brine (2 x 10 mL), dried (magnesium sulfate), and concentrated to give the crude product, which was purified by flash chromatography (silica, 20-30% ethyl acetate/hexane) to give the desired product as a white solid. MS (M+H) ⁺ 683.7, (MH) 681.6; IR (KBr) 1796 cnr *, 1734 cm-1, 1640 cm-1.

b)

(homochiral)

Deprotection and treatment of the product prepared in step (a) (53 mg, 0.078 mmol) according to the method of Example 19(c) gave 11 mg of the desired product as a white powder. MS (M+H) ⁺ 324.9, (MH) 323.1.

Example 38 (homochiral)

119

To a solution of the benzyl ester product prepared according to Example 1(c) (73.1 mg, 0.127 mmol) in dry tetrahydrofuran (2 mL) at -78 °C under nitrogen was added sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 131 μΐ, 0.131 mmol). The reaction mixture was stirred for 30 min at -78 °C and then 4-biphenylcarbonyl chloride (29 mg, 0.134 mmol) was added in one portion. The reaction mixture was then stirred for 5 min at -78 °C and then for 15 min at -15 °C. 1 N HCl (1 mL) was then added followed immediately by ethyl acetate (3 mL). The resulting biphasic solution was then stirred vigorously while being allowed to warm to room temperature. The organic phase was then separated, dried over magnesium sulfate, filtered, and concentrated to give a light yellow residue. Purification by flash chromatography (silica gel, 0-30% ethyl acetate in hexane) gave 28 mg of the desired product. IR (film) 1797 cm -1 ; MS 753.1 (M+H) ⁺ .

(homochiral)

Deprotection and treatment of the product prepared in step (a) (28 mg, 0.037 mmol) according to the method of Example 19(c) gave 11.8 mg of the desired product as a lyophilisate. IR (film)

120 ··· · · · ··

1788 cm-i; MS 395.1 (M+H) ⁺ , 393.3 (MH)-.

Example 39

a) Biphenylacetyl chloride

To a suspension of biphenylacetic acid (30 mg, 1.41 mmol) in methylene chloride (10 mL) was added dropwise oxalyl chloride (246 μΐ, 2.86 mmol) and one drop of dimethylformamide. The mixture was stirred at room temperature for 20 min. The solvent was evaporated and the residue was re-evaporated twice with toluene to give 310 mg of the title product as a yellow solid.

To a solution of the benzyl ester product prepared according to Example 1(c) (120 mg, 0.21 mmol) in tetrahydrofuran (3 mL) was added

121 • · · ··· · · ··· · ··· · ··· ·

Sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 315 μΐ, 0.31 mmol) was added dropwise at -78 °C. The resulting mixture was stirred for 2.5 hours at -78 °C. A solution of biphenylacetyl chloride (58 mg, 0.25 mmol) in tetrahydrofuran (1 ml) was then added. The reaction mixture was stirred at -78 °C for a further 2.5 hours and then left in the freezer (-50 °C) overnight. The reaction mixture was then partitioned between ethyl acetate (50 ml) and water (15 ml).

The organic layer was separated and washed with brine (20 mL), dried (magnesium sulfate), filtered, and concentrated to give the crude product, which was purified by flash chromatography (silica, 20-30% ethyl acetate/hexane) to give 22 mg of the desired product as a yellow solid. MS (M+H) ⁺ 767.1; IR (KBr) 1796 cm ¹ , 1730 cnr ¹ , 1640 cnr ¹ .

C)

NH»CF ₃ COOH

Deprotection and treatment of the product prepared in step (b) (40 mg, 0.052 mmol) according to the method of Example 19(c) gave 11 mg of the desired product as a white flaky powder. MS (M+H) ⁺ 409.2, (MH) - 407.2; IR (KBr) 1782 cm- ¹ ,

1684 cm~ ¹ , 1645 cm” ¹ .

Example 40

122 ···· · ··· · · • · · · ·«···«· ·

a) 4-benzylpiperidinylcarbonyl chloride

To a mixture of phosgene (3.8 mL of a 20% solution of phosgene in toluene, 7.13 mmol) was added 4-benzylpiperidine (0.5 g, 2.86 mmol). The mixture was stirred for 1 h at 0 °C. The reaction mixture was then partitioned between water (25 mL) and ethyl acetate (2 x 25 mL). The organic phase was washed with 1 N HCl (1 x 40 mL), saturated sodium chloride (1 x 50 mL), dried over sodium sulfate, and concentrated to give a yellow oil. Purification by flash column chromatography (silica gel, 0-10% ethyl acetate/hexane) yielded 0.47 g of the title product. IR (film) 1733.1 cm ^-1 .

b)

To a cold solution of the benzyl ester product prepared as in Example 1(c) (63 mg, 0.11 mmol) in methylene chloride (2 mL) at 0°C was added triethylamine (17 mg, 0.165 mmol) and 4-dimethylaminopyridine (6-8 crystals). 4-Benzylpiperidinylcarbonyl chloride (39 mg, 0.165 mmol) was then added and the mixture was stirred for 45 min at 0°C and then for 2.5 h at room temperature. The mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 68 mg of the desired product as a colorless oil. MS 774.2 (M+H) ⁺ , 772.4 (MH)-; IR (film) 1785.1, 1732.0, 1674.2, 1639.3 cm.

123

c) (homochiral)

Deprotection and treatment of the product prepared in step (b) (65 mg, 0.084 mmol) according to the method of Example 21(d) gave 39 mg of the desired product as a white lyophilisate. MS 416.2 (M+H) ⁺ , 414 (MH) - ; IR (KBr) 1784, 1657 cm 1 .

Example 41

(homochiral)

a) N-piperidinylcarhonyl chloride

To a mixture of phosgene (4.7 mL of a phosgene in toluene solution, 8.81 mmol) in methylene chloride (5 mL) at 0 °C was added piperidine (0.3 g, 3.52 mmol). The mixture was then stirred for 1 h at 0 °C. The reaction mixture was then evaporated in vacuo. The residue was suspended in ether, filtered, and the eluates were concentrated to give a yellow oil. Purification by flash chromatography (silica gel, 0-20% ethyl acetate/hexane) gave 0.162 g of the title product. IR (film) 1738.9 cm ^-1 ,

124 ··

O

To a cooled solution of the benzyl ester product prepared according to Example 1(c) (120 mg, 0.21 mmol) in methylene chloride (1 mL) at 0 °C was added triethylamine (32 mg, 0.314 mmol) and 4-dimethylaminopyridine (10-15 crystals). N-piperidinylcarbonyl chloride (46 mg, 0.314 mmol) was then added and the mixture was stirred for 16 hours at room temperature. The mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel, 0-20% ethyl acetate/hexane) gave 95 mg of the desired product as a colorless gum. MS 684.3 (M+H) ⁺ , 682.5 (MH)-; IR (film) 1783.9, 1731.0 cnr i .

c) (homochiral)

Deprotection and treatment of the product prepared in step (d) (65 mg, 0.089 mmol) according to the method of Example 21(d) gave 11 mg of the desired product as a colorless glass. MS 326.3 (M+H) ⁺ , 324.3 (MH) - .

125

Example 42

NH3CF,COOH

O (homochiral)

a) 1,2,3,4-tetrahydroisoquinolinylcarbonyl chloride

To a cold mixture of phosgene (5 mL of a 20% solution of phosgene in toluene, 9.4 mmol) in methylene chloride (5 mL) at 0 °C was added 1,2,3,4-tetrahydroisoquinoline (0.5 g, 3.76 mmol). The resulting mixture was stirred for 1 h at 0 °C. The reaction mixture was then evaporated in vacuo. The residue was suspended in ethyl ether, filtered, and the eluates were concentrated to a light pink oil. Purification by flash chromatography (silica gel, 0-10% ethyl acetate/hexane) gave 0.586 g of the desired product. IR (film) 1735.3 cm ^-1 .

II

To a cooled solution of the benzyl ester product prepared according to Example 1(c) (77 mg, 0.135 mmol) in methylene chloride (2 mL) at 0°C was added triethylamine (20 mg, 0.2 mmol) and 4-dimethylaminopyridine (8-10 crystals). 1,2,3,4-tetrahydroisoquinolinylcarbonyl chloride (39 mg, 0.2 mmol) was then added and the mixture was stirred at room temperature for 2.5 hours.

126 ·· ·· 4*4 ftft * « ft ί · · # » · • ftftft ft ftftft · · • n ftftft · · ft 9 ft » ft • ••ft ftft ·· · ·Τ ft

The mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 66 mg of the desired product as a colorless oil. MS 732.3 (M+H) ⁺ , 730.7 (MH) - ; IR (film) 1790.2, 1732.0, 1673.8, 1638.9 cm ^-1 .

c) (homochiral)

NH«CF ₃ COOH U h ₂ nn ² H £O ₂ H

Deprotection and workup of the product prepared in step (b) (65 mg, 0.089 mmol) according to the method of Example 19(c) gave 33 mg of the desired product as a white foam. MS 374.2 (M+H) ⁺ , 372.4 (MH) - ; IR (film) 1788.0, 1668.0 cm 1 .

Example 43

(homochiral)

a) ^O\— CH ₂ — O— C— N 1? 1 (Cp) CHT" O- C- n

about what ₂ -ch ₂

127 • · · · • · « · · · ·

To a solution of the benzyl ester product prepared according to Example 1(c) (92 mg, 0.16 mmol) in tetrahydrofuran (2 ml) at -78 °C was added dropwise sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 180 μΐ, 0.18 mmol). The resulting mixture was stirred for 1 hour at -78 °C. Phenoxyacetyl chloride (24 μΐ) was then added. The reaction mixture was then stirred for a further 20 minutes at -78 °C and then left overnight in a freezer (-50 °C). Analytical HPLC showed that the reaction was incomplete after this time. A further 24 μΐ of phenoxyacetyl chloride was added and the mixture was stirred for a further 3.5 hours at -78 °C. The reaction mixture was then quenched with water (10 ml). It was then extracted with ethyl acetate (3 x 20 ml). The organics were combined and washed with brine (2 x 10 mL), dried (magnesium sulfate), filtered, and concentrated to give the crude product, which was purified by flash chromatography (silica, 30% ethyl acetate/hexane) to give 65 mg of the desired product as a colorless oil. MS (M+H) ⁺ 707.1, (MH) 705.4; IR (KBr) 1798 cm-1, 1640 cm- ¹ .

(homochiral)

Deprotection and treatment of the product prepared in step (a) (63 mg, 0.089 mol) according to the method of Example 21(d) gave 31 mg of the desired product as a white powder. MS (M+H) ⁺ 349.0, (MH) - 347.2; IR (KBr) 1800 cm -1 , 1723 cm -1 , 1649 cm -1 • ·

128

Example 44

a) 4-biphenyl sulfonyl chloride

To a suspension of triphenylphosphine resin (1.58 g) in methylene chloride (10 ml) was added dropwise sulfuryl chloride (392 μΐ, 4.74 mmol) at 0 °C. Then a solution of 4-biphenyl sulfonic acid (400 mg, 1.58 mmol) and triethylamine (220 μΐ) in methylene chloride (5 ml) was added. The resulting mixture was stirred at room temperature for 6 hours and left at 5 °C for 3 days. It was then filtered and the filtrate was evaporated. The residue was coevaporated twice with toluene to give the crude product as a white solid. Purification of the crude product by chromatography (silica, methylene chloride) gave 180 mg of the title compound as a white solid.

To a solution of the benzyl ester product prepared according to Example 1(c) (95 mg, 0.17 mmol) in tetrahydrofuran (3 mL) was added:

129 • · · · · · · · « · · · sodium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 255 μΐ, 0.26 mmol) was added dropwise at -78 °C. The resulting mixture was stirred for 40 minutes at -78 °C. Then a solution of

4-biphenylsulfonyl chloride (64 mg, 0.26 mmol) in tetrahydrofuran (1 mL). The reaction mixture was then stirred for an additional 20 min at -78 °C and then left in the freezer (-50 °) overnight. After this time, analytical HPLC indicated the reaction was complete. The reaction mixture was then quenched with 1 N potassium hydrogen sulfate (20 mL). The mixture was then extracted with ethyl acetate (2 x 50 mL). The organic layers were combined, washed with brine (20 mL), dried (magnesium sulfate), filtered, and concentrated to give the crude product as a yellow oil (158 mg). Purification of the crude product by flash chromatography (silica, 30% ethyl acetate/hexane) gave 76 mg of the desired product as a colorless oil. MS (M+H) ⁺ 789.0.

Deprotection and treatment of the product prepared in step (b) (70 mg, 0.088 mmol) according to the method of Example 19(c) gave 17 mg of the title product as a white, flaky powder. MS (M+MeOH+H) ⁺ 463.2, (M+MeOH-H) - 461.5; IR (KBr) 1773 cm*,

1665 cm ^-1 , 1595 cm ¹ .

Example 45 trans-3-[3-[(aminoiminomethyl)amino]propyl]-2-oxo-4-(2-phenylethyl)-N-(phenylmethyl)-1-azetincarboxamide monohydrochloride

130

A solution of trans-4-(2-phenylethenyl)-3-[3-[Ν',N''-bis(carbobenzyloxy)guanidino]propyl]-2-azetidinone (130 mg, 0.2 mmol, prepared according to Example 3 of U.S. Patent No. 5,037,819 to Han) in tetrahydrofuran (1.5) was cooled to -78°C under argon. A 1 M solution of sodium bis(trimethylsilyl)amide (0.21 mL) in tetrahydrofuran was then added, and the mixture was stirred for 15 minutes. Benzyl isocyanate (40 mg, 0.3 mmol) was then added dropwise, the mixture was allowed to warm to room temperature, and stirred for 1 hour. The reaction mixture was then diluted with 10% aqueous potassium hydrogen sulfate (10 mL) and extracted with ethyl acetate (3 x 10 mL); The combined organics were washed with water (25 mL), brine (25 mL) and dried over sodium sulfate. The solution was filtered and the solvent removed in vacuo to give an oil. The residue was purified by flash chromatography (silica, ethyl acetate:hexane 2:3) to give 69 mg of the desired product as a colorless oil. MS (M+H) ⁺ 674.

b) trans-3-[3-(aminoiminomethyl)amino]propyl]-2-oxo131

-4-(2-phenylethyl)-N-(phenylmethyl)-1-acetate carboxamide, monohydrochloride

A solution of the product prepared in step (a) (67 mg, 0.1 mmol) in dioxane (1.5 mL) containing 1 N HCl (0.15 mL) and 10% palladium on carbon catalyst was stirred for 2 h under hydrogen. The reaction mixture was then filtered and lyophilized to give 66 mg of the title compound as a colorless solid; mp 145-154 °C (dec). MS (M+H) ⁺ 408. IR (KBr) 1761 cm.

Example 46 trans-3-[3-(amino iminomethyl)amino]propyl]-N-methyl-oxo-4-(2-phenylethyl)-1-azetinecarboxamide, monohydrochloride (racemate)

O

The desired product (80 mg) was prepared as a colorless oil by the method of Example 45(a) except that methyl isocyanate (23 mg,

132

0.5 mmol). MS (M+H) ⁺ 598.

b) trans-3-[3-[(aminoiminomethyl)amino]propyl]-N-methyl-2-oxo-4-(2-phenylethyl)-1-azet incarboxamide, monohydrochloride

Deprotection and treatment of the product prepared in step (a) (77 mg, 0.13 mmol) according to the method of Example 45(b) gave 42 mg of the title product as a colorless solid; mp 138-146 °C (dec). MS (M+H) ⁺ 332. IR (KBr) 1761 cm ^-1 .

Example 47

[chiral (+)]

a) [chiral (-)] /-Λ ¹¹ (O)-CH2-OCN

About JI (θ)-CH2-O-C-N

CH=CH —<2>

Trans-4-(2-phenylethenyl)-3-[3-[N',N''-bis(carbobenzyloxy) guanidino]propyl]-2-azetidinone is separated into an enantiomerically pure (-) isomer and a (+) isomer on a Chiralpak-ADR preparative column using a 30% 2-propanol/hexane mixture as eluent.

• ·

133

b)

A solution of chiral (-)-trans-4-(2-phenylethenyl)-3-[3-[N',N''-bis(carbobenzyloxy)guanidino]propyl]-2-azetidinone (216 mg, 0.40 mmol) in tetrahydrofuran (2.0 mL) was cooled to -78 °C under argon. A 1 M solution of bis(trimethylsilyl)amide (0.44 mL) in tetrahydrofuran was then added and the mixture was stirred for 15 min. Acetyl chloride (32.2 mg, 0.41 mmol) was then added dropwise and the mixture was warmed to room temperature and stirred for 1 h. The reaction mixture was then diluted with 10% aqueous potassium hydrogen sulfate (10 mL) and extracted with ethyl acetate (3 x 10 mL); The combined organics were then washed with water (25 mL), brine (25 mL) and dried over sodium sulfate. The solution was then filtered and the solvent evaporated to give an oil. Purification of the residue by flash chromatography (silica, ethyl acetate:hexane 1:3) afforded 150 mg of the desired product as a colorless oil. MS (M+H) ⁺ 583.

C)

( + )]

A solution of the product prepared in step (b) (140 mg, 0.258 mmol) in dioxane (3.5 mL) containing 1 N HCl (0.3 mL) and 10%

134 • * • · ♦ · · · · · ··· · ··· · ·

palladium on carbon catalyst (60 mg) was stirred under hydrogen atmosphere for 1 hour. The reaction mixture was then filtered and lyophilized to give 64 mg of the desired product as a colorless solid. MS (M+H)* 317; IR (KBr) 1782 cm ¹ ; [q]d=+18° (c=1, methanol).

[chiral (+)]

The desired product was prepared according to the method of Example 47(b) and (c) using chiral (+)-trans-4-(2-phenylethenyl)-3-[3-[N',N''-bis(carbobenzyloxy)guanidino)propyl]-2-azetidinone.

(racemate)

and)

About <2>

135

A solution of cis-4-(2-phenylethenyl)-3-[3-[Ν',N''-bis(carbobenzyloxy)guanidino]propyl]-2-azetidinone (300 mg, 0.555 mmol, prepared according to Example 3 of US Patent 5,037,819 to Han) in tetrahydrofuran (2.5 mL) was cooled to -78°C under argon. A 1 M solution of sodium bis(trimethylsilyl)amide in tetrahydrofuran (0.8 mL) was then added and the mixture was stirred for 15 minutes. 4-Biphenylcarbonyl chloride (180 mg, 0.832 mmol) was then added dropwise, the mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction mixture was then diluted with 10% aqueous potassium hydrogen sulfate (15 mL) and extracted with ethyl acetate (3 x 15 mL); The combined organics were washed with water (25 mL), brine (25 mL) and dried over sodium sulfate. The solution was filtered and evaporated to an oil. The residue was purified by flash chromatography (silica, ethyl acetate:hexane, 1:4) to give 320 mg of the desired product as a colorless solid. MS (M+H) ⁺ 721.

b)

A solution of the product prepared in step (a) (300 mg, 0.48 mmol) in dioxane containing aqueous 1 N HCl (0.65 mL) and 10% palladium on carbon catalyst (150 mg) was stirred for 2 hours under hydrogen atmosphere. The reaction mixture was then lyophilized to give 178 mg of the desired product as a colorless solid. MS (M+H) ⁺ 455. IR (KBr) 1782 cm.

• ·

136

Example 50

and)

A solution of chiral trans-4-(2-phenylethenyl)-3-[3-[N',N-bis(carbobenzyloxy)guanidino]propyl]-2-azetidinone (264 mg, 0.49 mmol, in tetrahydrofuran (2.5 ml) under argon was cooled to -78 °C. A 1 M solution of sodium bis(trimethylsilyl)amide in tetrahydrofuran (0.74 ml) was then added and the mixture was stirred for 15 minutes. 4-Biphenylcarbonyl chloride (163 mg, 0.75 mmol) was then added dropwise, the mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction mixture was then diluted with 10% aqueous potassium hydrogen sulfate (15 ml) and extracted with ethyl acetate (3 x 15 ml); The combined organics were washed with water (25 mL), brine (25 mL) and dried over sodium sulfate. The solution was filtered and evaporated to an oil. The residue was purified by flash chromatography to give 290 mg of the desired product as a colorless solid. MS (M+H) ⁺ 721 .

• · ··· ···«··· • · · · · · · ·

137

b)

Deprotection and workup of the product prepared in step (a) (280 mg, 0.4 mmol) according to the method of Example 49(b) gave 172 mg of the desired product as a colorless solid. MS (M+H) ⁺ 455. IR (KBr) 1782 cm-1; [α] 22 = +12° (c = 1, methanol).

Example 51

O

The desired product (134 mg) was prepared as a colorless solid by the method of Example 49(a) using (2-naphthyloxy)acetyl chloride instead of 4-biphenylcarbonyl chloride. MS (M+H) ⁺ 725.

138 • · ·

Deprotection and treatment of the product prepared in step (a) (125 mg, 0.17 mmol) according to the method of Example 49(b) gave 76 mg of the desired product as a colorless solid. MS (M+H) ⁺ 459. IR (KBr) 1780 cm -1 .

Example 52 (homochiral)

O

The desired product (216 mg) was prepared as a colorless solid by the method of Example 50(a) using (2-naphthyloxy)acetyl chloride instead of 4-biphenylcarbonyl chloride. MS (M+H) ⁺ 725.

139

Deprotection and treatment of the product prepared in step (a) (200 mg, 0.276 mmol) according to the method of Example 49(b) gave 108 mg of the desired product as a colorless solid. MS (M+H) ⁺ 459. IR (KBr) 1780 cm - ¹ , [α] 22 = + 18° (c = 1, methanol).

Example 53

A solution of the carboxylic acid derivative prepared according to Example 1(b) (140 mg, 0.288 mmol) in tetrahydrofuran (2.5 mL) was cooled to -20°C under argon and then N-methylmorpholine (32.1 mg, 0.317 mmol) was added. 2 M dimethylamine (1.1 eq.) in tetrahydrofuran was then added and then benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (140 mg, 0.317 mmol) was added. The reaction mixture was then stirred for 4

140 « » hours 24 hours at -20 °C, then poured into 5% potassium hydrogen sulfate solution and extracted with ethyl acetate.

The ethyl acetate extract was then washed with water, brine and dried over sodium sulfate. The solvents were evaporated and the crude residue was purified by chromatography on silica using ethyl acetate as eluent to give 56 mg of the desired product as a colorless solid. MS (M+H) ⁺ 510.

A solution of the product prepared in step (a) (48 mg, 0.094 mmol) in tetrahydrofuran (0.4 mL) was cooled to -78 °C under argon. Sodium bis(trimethylsilyl)amide (1 M, 1.5 eq.) was then added and the mixture was stirred for 30 min. (R)-(-)-1-naphthenyl)-ethyl isocyanate (27.2 mg, 0.141 mmol) was then added. The mixture was stirred for 30 min at -78 °C, then allowed to warm to room temperature and stirred for 4 h. The reaction mixture was then poured into 5% potassium hydrogen sulfate and extracted with ethyl acetate. The ethyl acetate extract was then washed with water, brine and dried over sodium sulfate. The solvents were evaporated and the crude residue was purified by chromatography on silica using ethyl acetate:hexane (3:2) to give 25 mg of the desired product as a colorless glass. MS (M+H) ⁺ 707.

141 · « · ·

c) ·« *« ·· « · * * • · * * · • « · · · • · · · ·»·· ·· ··

A solution of the product prepared in step (b) (24 mg, 0.03 mmol) in dioxane (1 mL) containing HCl (1.5 eq.) was stirred under hydrogen atmosphere with 10% palladium on carbon catalyst (12 mg) for 2 h. The reaction mixture was then filtered and lyophilized to give a colorless solid (methanol).

then 14 mg of the desired product mass is obtained. MS (M+H) ⁺ 431; [α]d=+12° (c=1,

Example 54

HN (CH ₂ ) 2 “ N- C— O— C (CH1 I

CH(CH ₃ ) ₂ CH(CH ₃ ) ₂ (homochiral)

O

-N^°-C(CH3)3 CH(CH ₃ ) ₂

3)3

A mixture of di-tert-butyl dicarbonate (1.51 g, 6.9 mmol) and triethylamine (0.7 g, 6.9 mmol) in anhydrous tetrahydrofuran (3 mL) was added over 20 minutes to a solution of N,N'-diisopropylethylediamine (1.0 g, 6.9 mmol) in anhydrous tetrahydrofuran (2 mL). The mixture was stirred for 2 hours at room temperature. The mixture was then filtered and washed with methylene chloride. The filtrate and washings

142 • · ··· ··» >··· ^·

b)

Cl— C—N (CH ₂ ) ₂ -NC O—C(CH ₃ ) ₃

CH(CH ₃ ) ₂ CH(CH ₃ ) ₂

To a mixture of phosgene (0.24 ml of a 20% solution of phosgene in toluene, 0.45 mmol) in methylene chloride (1 ml) was added at 0 °C the product prepared in step (a) (44 mg, 0.18 mmol) and then triethylamine (25 μΐ, 0.18 mmol) was added. The resulting mixture was stirred for 1 hour at 0 °C. The reaction mixture was then evaporated in vacuo. Purification of the residue by flash chromatography (silica gel 0-10% ethyl acetate/hexane) gave about 30 mg of the desired product as a colorless oil. IR (neat) 1732.0, 1694.5 cm ^{- 1} .

C)

CO2CH2

About II

N—(CH ₂ ) ₂ -NC—OC(CH ₃ ) ₃

I o CH(CH ₃ ) ₂ CH(CH ₃ ) ₂

To a solution of the benzyl ester product prepared in Example 1(c) (40 mg, 0.07 mmol) in methylene chloride (1 mL) were added triethylamine (15 μΐ, 0.104 mmol) and dimethylaminopyridine (10-12 crystals), followed by the addition of the acid chloride derivative prepared in step (b) (25 mg, 0.084 mmol). The resulting mixture was stirred for 48 hours, then evaporated in vacuo and purified by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) to give the title compound.

143 gives 21 mg of the desired product as a colorless oil. MS

843.5 (M+H) ⁺ , 841.8 (MH)"; IR (film) 1785.1, 1733.1, 1681.7, 1640.9, cm ^-1 .

d)

NH

(homochiral)

About

N— (CH ₂ ) ₂ -n °-C(CH ₃ ) ₃

CH(CH ₃ ) ₂ CH(CH ₃ ) ₂

To a solution of the product prepared in step (c) (21 mg,

0.025 mmol) in 1,4-dioxane (5 ml) containing 1 N HCl (25 μΐ, 0.025 mmol) was added as a catalyst, 10% palladium on carbon (20 mg, wet type). Hydrogen was then bubbled through the solution for 4 hours. The reaction mixture was then filtered through a pad of Celite ^R , which was then washed again with 1,4-dioxane (10 ml) and water (15 ml).

The combined eluates were then lyophilized. The white lyophilizate was dissolved in water and passed through a polyvinylpyrrolidone bed using water as eluent. The eluates were lyophilized again to give 12 mg of the desired product as a white lyophilizate. MS 485.3 (M+H) ⁺ , 483.5 (MH) - ; IR (KBr) 1778.0, 1665.0 cm -1 .

Example 55 (homochiral

NH- _2CF3COOH

Λ

H ₂ NN ² H _aC0 ₂ H

144 • · · · · · · · · · · · · · · · · · · · · · · «··· • · · · · · · · · · · · · · *

and)

To the phosgene mixture (3.5 ml of a 20% solution of phosgene in toluene,

6.6 mmol) in methylene chloride (2 ml) at 0 °C is added

1-phenethylpiperazine (0.5 g, 2.63 mmol) and then triethylamine (0.37 ml, 2.63 mmol). The resulting mixture was stirred for 2 hours at 0 °C and then evaporated in vacuo. The residue was suspended in ether, filtered and the eluates were concentrated to give a creamy solid. Purification by flash chromatography (silica gel,

0-10% ethyl acetate/hexane) gave 32.2 mg of the desired product as a crystalline white solid. IR (film) 1729.3 O

To a solution of the benzyl ester product prepared according to Example 1(c) (129 mg, 0.23 mmol) in methylene chloride (3 mL) were added triethylamine (47 μΐ, 0.34 mmol) and dimethylaminopyridine (10-12 crystals) followed by the addition of the acid chloride derivative prepared in step (a) (86 mg, 0.34 mmol). The resulting mixture was stirred at room temperature for 5 hours and then evaporated in vacuo to a light yellow paste. Purification by flash chromatography (silica gel, 0-35% ethyl acetate/hexane) gave 120 mg of the desired product as a colorless oil. MS 789.4 (M+H) ⁺ , 787.7 (MH)-; IR (film) 1785.5, 1732.1, 1679.1, 1639.4, cm-1.

C)

NH·2 CF ₃ COOH

Λ h ₂ n N' ² H (homochiral) ^co ₂ ho

145

To a solution of the product prepared in step (b) (118 mg, 0.15 mmol) in 1,4-dioxane (8 mL) containing 1 N HCl (0.18 mL, 0.18 mmol) was added the catalyst, 10% palladium on carbon (60 mg, wet type). Hydrogen was then bubbled through the solution for 1 to 1.5 hours. The reaction mixture was then filtered through a pad of Celite ^R , which was washed again with 1,4-dioxane (10 mL) and water (15 mL). The combined eluates were lyophilized to give 77 mg of a white lyophilisate. Purification by HPLC (reverse phase, methanol, water, trifluoroacetic acid) gave 52 mg of the desired product as a white lyophilisate. MS 431.2 (M+H) ⁺ , 429.3 (MH) ^- ; IR (KBr) 1790.0, 1678.0 cm-1.

Example 56

H

(homochiral)

A solution of 1-(2-pyrimidyl)piperazine dihydrochloride (4.0 g, 16.9 mmol) in 1 N sodium hydroxide saturated with sodium chloride (40 mL) was extracted with ethyl acetate (2 x 30 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated to give 2.6 g of 1-(2-pyrimidyl)piperazine.

To a solution of phosgene (25 ml, 20% solution in toluene, 47.3 mmol) in methylene chloride (15 ml) was added dropwise over solid sodium bicarbonate (3 g) over 3 minutes under a nitrogen atmosphere and at room temperature a solution of

146 ··· 0 0 0 0 0 0 0

0 0 0 0 · · 0 · · · • · · * · 0000000 · · ··· » 0 t * · · ···· »· 00 « 00 ·0· l-(2-pyrimidyl)piperazine (2.6 g) in methylene chloride (5 ml). The resulting solution was then stirred vigorously for 10 minutes, then filtered through a sintered funnel, and the collected solids were washed with methylene chloride (2x5 ml). The combined eluates were concentrated in vacuo to give a white solid. Recrystallization of this solid from methylene chloride/hexane gave 3 g of the desired product as a white solid. IR (film) 1735 cm ^-1 .

O

To a solution of the benzyl ester product prepared in Example 1(c) (1.56 g, 3.23 mmol) in methylene chloride (15 mL) under nitrogen at room temperature was added the acid chloride prepared in step (a) (1.11 g, 4.8 mmol), triethylamine (700 μΐ, 5.0 mmol) and dimethylaminopyridine (200 mg, 1.64 mmol). The reaction mixture was stirred for 7 hours at room temperature, then diluted with hexane (5 mL) and then loaded onto the top of a silica gel column (wetted with methylene chloride) and purified by flash chromatography (0 to 30% ethyl acetate/methylene chloride) to give 1.6 g of the derivative product as a white foam. MS 763.2 (M+H) ⁺ , 761.7 (MH)-; IR (KBr) 1788 cm-1.

c) (homochiral)

O

147

Deprotection and treatment of the product prepared in step (b) (1.6 g, 2.1 mmol) according to the method of Example 21(d) gave 854 mg of the desired product as a white lyophilized solid. MS 409.2 (M+H) ⁺ , 407.5 (MH) - ; IR (KBr) 1777 cm 1 .

Analysis: calculated for Ci7H25NsOa.1.0 HCl.1.54 H2O:

C, 43.20; H, 6.84; N, 23.71; O, 18.75; Cl, 7.50; found:

C, 43.31; H, 6.59; N, 23.09; 0 (not determined);

Cl, 7.06.

Example 57

NH»2HCl χ

H ₂ NN (homochiral)

and)

,co ₂ h /-K Jk ^ ^CH 2

I ΊΟ (H3O3—C— 0-cN N \__y dd/NH \

co-ch ₂ II o

To a solution of N-carbobenzyloxy-L-phenylalanine (0.5 g, 1.6 mmol) in anhydrous methylene chloride (5 mL) was added triethylamine (0.22 mL, 1.6 mmol) and pyridinebenzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (0.83 g, 1.6 mmol), followed by the addition of tert-butyl-1-piperazinecarboxylate (0.3 g, 1.6 mmol). The resulting mixture was stirred at room temperature for 2 h. The mixture was then diluted with methylene chloride (30 mL) and washed with 1 N HCl (1 x 25 mL), saturated sodium bicarbonate (1 x 25

148 mL) and saturated sodium chloride (1 x 25 mL). The organic phase was dried over sodium sulfate and concentrated to give a light yellow oil. Purification by flash chromatography (silica gel, 0-20% ethyl acetate/hexanes) gave 554 mg of the desired product as a white foam. IR (film) 1698.2, 1649.6 cm ^-1 .

b) <-\

HN N i / NH ^x co-ch ₂

To a solution of the product prepared in step (a) (550 mg, 1.14 mmol) in anhydrous methylene chloride (3 mL) at 0 °C was added trifluoroacetic acid (3 mL). The mixture was then warmed to room temperature and stirred for 1.5 hours. The mixture was concentrated to give a colorless oil. This oil was dissolved in water, the pH was adjusted to 12-13 with sodium hydroxide (50% solution) and extracted with ethyl acetate (3 x 50 mL). The organic phase was dried over sodium sulfate and concentrated to give 0.48 g of the desired product as a pale yellow oil. MS 368.2 (M+H) ⁺ .

To a mixture of phosgene (1.11 ml of a 20% solution of phosgene in toluene, 2.1 mmol) in methylene chloride (3 ml) at 0 °C was added the product prepared in step (b) (0.322 g, 0.84 mmol) in

149 « · • · • · • · t · « · methylene chloride (3 ml) and then triethylamine (0.12 ml, 0.84 mmol) was added. The resulting mixture was stirred for 1 hour at 0 °C. The reaction mixture was then evaporated in vacuo. The residue was suspended in ether, filtered, and the eluates were concentrated to give a yellow oil. Purification by flash chromatography (silica gel, 0-20% ethyl acetate/hexane) gave 0.243 g of the desired product as a colorless liquid.

d)

To a solution of the benzyl ester product prepared in Example 1(c) (100 mg, 0.175 mmol) in methylene chloride (3 mL) were added triethylamine (37 µL, 0.262 mmol) and dimethylaminopyridine (10-12 crystals), followed by the addition of the acid chloride prepared in step (c) (120 mg, 0.262 mmol).

The resulting mixture was stirred for 4 hours at room temperature and then evaporated in vacuo. Purification of the crude product by flash chromatography (silica gel, 0-35% ethyl acetate/hexane) gave 131 mg of the desired product as a colorless oil. MS 966.4 (M+H) ⁺ , 964.6 (MH) - ; IR (film) 1788.0, 1738.3, 1677.1, 1637.0 cm -1 .

e) (homochiral)

NH»2HC1 H ₂ N^N^^A

O

every ₂ hours

A/“ ² nh ₂ w

150 in · • · ·

To a solution of the product prepared in step (d) (127 mg, 0.132 mmol) in 1,4-dioxane (8 mL) containing 1 N HCl (0.26 mL, 0.264 mmol) was added the catalyst, 10% palladium on carbon (65 mg, wet type). Hydrogen was then bubbled through the solution for 1 to 1.5 hours. The reaction mixture was then filtered through a pad of Celite ^R , which was then washed with 1,4-dioxane (10 mL) and water (15 mL). The combined eluates were lyophilized to give 64 mg of the desired product as a pale yellow lyophilisate. MS 474.2 (M+H)+, 472.4 (MH)-; IR (KBr) 1786.0, 1730.0, 1647.0 cm ¹ .

Example 58 (homochiral)

NH»HCl h ₂ n ^z ^'n ^x ^- ^x

H

O—C(CH ₃ ) ₃

and)

HN-(CH ₂ ) ₂ ch ₃

C(CH ₃ ) ₃

To a solution of N,N ¹ -dimethylethylenediamine (500 mg, 5.67 mmol) in tetrahydrofuran (35 ml) was added a solution of di-tert.butyldicarbonate (1.24 g, 5.67 mmol) and triethylamine (790 μΐ, 5.67 mmol) in tetrahydrofuran (15 ml). The reaction mixture was stirred for 4 days at room temperature. The mixture was then filtered and the filtrate was concentrated to give the crude product as a colorless oil. Purification by flash chromatography (10% 2 N ammonia in methanol/methylene chloride) gave 362 mg of the desired product as a colorless oil. IR (film) 1694 cm ^-1 .

• · about

II

C(CH ₃ ) ₃ cl— c— N (CH ₂ ) 2“ θ—

b)

O

II

151 ch ₃ ch ₃

To a solution of phosgene in toluene (1.4 ml, 20%) was added at 0 °C a mixture of the product prepared in step (a) (250 mg) and triethylamine (278 μΐ) in methylene chloride (3 ml). The resulting mixture was stirred for 5 hours at 0 °C. Anhydrous ether (10 ml) was added and the solid was filtered off. The filtrate was evaporated to give the crude product as an orange oil, from which 313 mg of the desired product was obtained as a colorless oil by flash chromatography (20-30% ethyl acetate/hexane). IR (film) 1740 cm ^-1 , 1694 cm ^-1 ,

C)

O

Γ o—C(CH ₃ )3

To a solution of the benzyl ester product prepared according to Example 1(c) (70 mg) in methylene chloride (2 ml) were added triethylamine (25 μΐ) dimethylaminopyridine (15 mg) and a solution of the acid chloride prepared in step (b) (45 mg, 0.18 mmol) in methylene chloride (1 ml). The resulting mixture was stirred overnight at room temperature. The reaction mixture was then quenched with 1 N sodium hydrogen sulfate (15 ml). This mixture was extracted with ethyl acetate (2 x 30 ml). The organics were combined and washed with brine (10 ml), dried over magnesium sulfate and concentrated to give 101 mg of crude product as a yellow oil. Purification by flash chromatography (30-50%

152 • · • » ethyl acetate/hexane) gives 77 mg of the desired product as a colorless oil. IR (film) 1786 cm ^-1 , 1733 cm ^-1 , 1681 cm ^-1 , 1639 cm ^-1 .

d)

NH»HC1 (homochiral)

H ₂ NT'> ^cq 2H ^H fj ff — ^ϊ; γ^~ ^(CHz} c— O— c (CH ₃ ) 3

_About ^CH3CH3

A mixture of the product prepared in step (c) (74 mg, 0.094 mmol), 1 N HCl (94 μΐ) and palladium-on-carbon catalyst (10%, 19 mg) in dioxane (2 mL) was stirred under hydrogen atmosphere (hydrogen balloon) for 1 h at room temperature. The reaction mixture was then filtered through a pad of Celite ^R and lyophilized to give 44 mg of the desired product as a white solid. MS 429.2 (M+H) ⁺ , 427.5 (MH); IR (KBr) 1784, 1663 cm.

Example 59 (homochiral)

To a solution of phosgene (3 mL, 20% solution in toluene, 5.6 mmol) in toluene (3 mL) under nitrogen at room temperature was added

153 a solution of N-methyl-4-phenylbenzylamine [158 mg, 0.8 mmol, prepared according to Dahn et al., Chim. Acta, 35, 1348-1358, (1952)] in toluene (3 ml) was added and then triethylamine (200 μΐ, 1.43 mmol) was added. After stirring at room temperature for 30 minutes, the solvents were removed in vacuo and the residue was purified by flash chromatography (silica gel, 100% methylene chloride) to give 136 mg of the desired product as an oily residue. IR (film)

To a solution of the benzyl ester product prepared as in Example 1(c) (145 mg, 0.25 mmol) in methylene chloride (3 mL) under nitrogen at room temperature was added the acid chloride prepared in step (a) (100 mg, 0.38 mmol), triethylamine (53 μΐ, 0.38 mmol) and dimethylaminopyridine (10 mg, 0.08 mmol). The reaction mixture was then stirred at room temperature for 6 hours, then diluted with hexane (1 mL) and loaded onto the top of a silica gel column (wetted with hexane) and purified by flash chromatography (0-20% ethyl acetate in hexane) to give 148 mg of the desired product as a light brown wax. MS

796.5 (M+H) ⁺ , 794.7 (MH) - ; IR (Film) 1786 cm-i.

C)

154 ft · · *

AND

Deprotection and purification of the product prepared in step (b) (148 mg, 0.186 mmol) according to the method of Example 19(c) gave 33 mg of the desired product as a white foam. MS 438.2 (M+H) ⁺ , 436.4 (MH) - ; IR (KBr) 1788.0, 1699.0 cm 1 .

Example 60

NH«HC1 h ₂ n^· _k CO ₂ H

N— C— O— C(CH ₃ ) ₃ H

a) í?

Cl—C-N,

O

II

IjT—c—o—C(CH ₃ ) ₃ H

To a stirred solution of 3-tert-butoxycarbonylaminoazetidine [250 mg, 1.45 mmol, prepared as described in Arimoto et al., J.Antibiot., 39(9), 1243-56, (1986)] and triethylamine (222 μΐ, 1.6 mmol) in methylene chloride (5 mL) was added phosgene (20% in toluene) at 0 °C. After 1 h, the reaction mixture was concentrated and the crude product was purified by silica gel chromatography to give 90 mg of the desired product.

The acid chloride prepared in step (a) (119 mg, 0.506 mmol) and the benzyl ester product prepared according to Example 1(c)

155 « · • · (193 mg, 0.337 mmol) was dissolved in methylene chloride (2.5 mL). Triethylamine (71 μΐ, 0.506 mmol) was then added followed by dimethylaminopyridine (8 mg, 0.067 mmol). After 12 h, the reaction mixture was concentrated and the crude product was purified by silica gel chromatography to give 180 mg of the desired product.

c) (homochiral)

NH-HC1

The product prepared in step (b) (80 mg, 0.104 mmol) was dissolved in 1,4-dioxane (1.0 mL) and water (0.10 mL). 1 N HCl (104 μΐ, 0.104 mmol) was then added followed by the catalyst, 10% palladium on carbon (16 mg). A hydrogen atmosphere was then introduced from the balloon. After stirring for 40 min at room temperature, the reaction mixture was diluted with water:1,4-dioxane (1:1) and filtered. Lyophilization of the filtrate gave 47 mg of the desired product. IE (KBr) 1792 cnr i.

Example 61 (homochiral)

156

and)

Ζθ)—ch ₂ —o—c-CH7- o- c-

what ₂ ch ₂ —<o>

_In the

_{NH2.CF3COOH ​}

To a stirred solution of the product prepared according to the method of Example 60(b) (100 mg, 0.13 mmol) in methylene chloride at 0°C was added dropwise trifluoroacetic acid (0.20 mL). The reaction mixture was then stirred at room temperature. After 40 minutes, the reaction mixture was concentrated in vacuo to give 120 mg of the desired product.

Deprotection and treatment of the product prepared in step (a) (120 mg, 0.153 mmol) according to the method of Example 60(c) gave 51 mg of the desired product. IR (KBr) 1788 cm ^-1 .

Example 62

NH»HC1

O

157

a) • · ··· ·····*· • · · · · · · «····· · · 0 ·· /δ\-ch ₂ — ocn

ABOUT JI <g>— CHJ-O-C-N

co ₂ ch ₂ cooch ₃

To a solution of the benzyl ester product prepared according to Example 1(c) (81.5 mg, 0.142 mmol) in dry tetrahydrofuran (5 mL) under nitrogen at -78 °C was added dropwise over 5 min 1 N 1,1,1,3,3,3-hexamethyldisilazane in tetrahydrofuran (143 μΐ, 0.14 mmol). After warming to -20 °C the reaction mixture was stirred for 30 min and methyl-(S)-(-)-2-isocyanato-3-phenylpropionate (29.2 mg, 0.14 mmol) dissolved in tetrahydrofuran was added dropwise. After stirring for one or more hours, the reaction solution was allowed to warm to 0°C, poured into a solution of potassium hydrogen sulfate (30 mL, pH adjusted to 3.5) containing crushed ice, and then extracted with ethyl acetate (3 x 15 mL). The combined organics were washed with water and brine, and finally dried over sodium sulfate. Concentration of the filtrate in vacuo gave 102 mg of crude product as a pale yellow oil. Purification by flash chromatography on silica gel using ethyl acetate/hexane (1:1) as eluent gave 90.8 mg of the desired product as a colorless oil. IR (film) 1780 cm- ¹ , 1743 cm ^-1 , and 1639 cm-1 ; MS 788.8 (M+H) ⁺ .

b)

NH*HC1

158

The product prepared in step (a) (0.12 mmol) was dissolved in dioxane (5 ml). Then, the catalyst, 10% palladium on carbon (40 mg), then 1 N HCl in ether (120 μΐ) were added and hydrogen was bubbled through the reaction suspension for 90 minutes at a constant flow. After the reaction was confirmed by TLC, the excess hydrogen was removed from the reaction medium using a nitrogen stream and then the catalyst was filtered off. The mixture was filtered through a HyfloSuperCel ^R pad, which was washed first with dioxane and then with a dioxane/water mixture to obtain a clear filtrate. This mixture was concentrated in vacuo and the residue was lyophilized to obtain 45 mg of the desired product as a white powder. IR (KBr) 1769 cm' ¹ , 1674 cm- ¹ and 1632 cm- ¹ ; MS 420.1 (M+H) ⁺ .

Example 63 (homochiral)

To a solution of N-carbobenzyloxy-D-phenylalanine (0.84 g, 2.68 mmol) in anhydrous methylene chloride (10 mL) was added triethylamine (0.37 mL, 2.68 mmol) and pyridine-benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (0.84 g, 2.68 mmol), followed by tert-butyl-1-piperazinecarboxylate (0.5 g, 2.68 mmol). The mixture was stirred at room temperature for 5 h, then methylene chloride (20 mL) was added and the mixture was

159 »· · · · · · I • · ftftft ft • ftftft · ftftft · • · ftftft ft······ washed with 1 N HCl (1 x 25 mL), saturated sodium bicarbonate (1 x 25 mL) and saturated sodium chloride. The organic phase was then dried over sodium sulfate and concentrated to give the crude product as a pale yellow oil. Purification by flash chromatography (silica gel, 0-30% ethyl acetate/hexane) gave 1.14 g of the desired product as a white foam.

b)

O

O

To a solution of the product prepared in step (a) (220 mg, 0.48 mmol) in anhydrous methylene chloride (2 mL) at 0 °C was added trifluoroacetic acid (2 mL). The mixture was then warmed to room temperature and stirred for 1.5 hours. The mixture was then concentrated to a colorless oil which was taken up in a solution of 1 N HCl in ether (0.48 mL) and the mixture was stirred vigorously. The resulting suspension was concentrated to give 230 mg of the hydrochloride salt of the desired product. MS 368.2 (M+H) ⁺ .

To a mixture of phosgene (0.36 mL of 20% phosgene in toluene solution, 0.68 mmol) and sodium bicarbonate (300 mg) in methylene chloride (4 mL) was added the product prepared in step (h) (110 mg, 0.27 mmol). The mixture was stirred for 3 hours at room temperature.

160 ·· ··· · · · • ·> · · ··· · · • · · · · ····*· · room temperature and then filtered, and by concentrating the eluted fractions, 165 mg of the desired product is obtained in the form of a clear gel.

IR (film) 1707.4, 1645.6 cm-».

To a solution of the benzyl ester product prepared in Example 1(c) (86 mg, 0.15 mmol) in methylene chloride (3 mL) were added triethylamine (31 μΐ, 0.23 mmol) and dimethylaminopyridine (10-12 crystals), followed by the acid chloride prepared in step (c) (99 mg, 0.23 mmol). The mixture was stirred at room temperature for 1 hour, then concentrated and purified by flash column chromatography (silica gel, 0-50% ethyl acetate/hexane) to give 90 mg of the desired product as a colorless oil. MS 966.5 (M+H) ⁺ , 964.7 (MH) ^- ; IR (film) 1786.4, 1727.5, 1639.5 cm-».

E)

Deprotection and treatment of the product prepared in step (d) (89 mg, 0.092 mmol) according to the method of Example 55(c) gave 68 mg of the desired product as a white lyophilisate. MS

161 • « • · · · · · · ···· · · « · · · • · ··· ········

474.3 (M+H)

Example 64

472.6 (M-H)~; IR (KBr) 1790.0, 1670.0 curi

and)

NH H ₂ N^

H

COOH (homochiral) _χ Ν— so ₂ —N (CH ₃ ) 2 O if (H,C) ₃ O-0- Ct^N-SCh—N (CH ₃ ) 2

To a solution of N-(tert-butoxycarbonyl)piperazine (600 mg, 3.22 mmol) in methylene chloride (15 mL) was added diisopropylethylamine (560 μΐ), dimethylaminopyridine (33 mg) and dimethylsulfamoyl chloride (462 mg, 3.22 mmol). The resulting mixture was stirred overnight at room temperature. The reaction was quenched by pouring in 1 N HCl (20 mL). This mixture was then extracted with ethyl acetate (2 x 100 mL). The extracts were combined and washed with brine (2 x 20 mL), dried over magnesium sulfate and concentrated to give 0.93 g of crude product as a white solid, which was used without further purification.

b) ^HC1 * ΠΝχ / ^N ~ ^S °2~ ^N (CH ₃ ) 2

A mixture of the product prepared in step (a) (0.60 g, 2.0 mmol), trifluoroacetic acid (15 mL) and methylene chloride (30 mL) was stirred at room temperature for 2 h. After this time, TLC showed the reaction to be complete. The residue was dissolved in a minimum amount of methylene chloride and then 1 N HCl/ether (2.0 mL) was added.

162 ml) and anhydrous white powder (M+H) ⁺ 194.1.

ether (20 ml). The product was filtered off to give 430 mg of the desired product in the form of MS.

C)

Cl— C- N\ SO ₂ —N (CH ₃ )

To a solution of phosgene (2.1 ml, 20% in toluene) in methylene chloride (20 ml) was added sodium bicarbonate (3.0 g) and then the product prepared in step (b) (300 mg, 1.3 mmol) was added. The resulting reaction mixture was stirred for 40 minutes at room temperature. After this time, TLC showed the reaction to be complete. The reaction was then terminated by filtering off the sodium bicarbonate. Evaporation gave 330 mg of the desired product. IR (film) 1738 cm ^-1 .

To a solution of the benzyl ester product prepared according to Example 1(c) (100 mg) in methylene chloride (1 mL) were added diisopropylethylamine (35 µL, 0.20 mmol), dimethylaminopyridine (23 mg) and a solution of the acid chloride prepared in step (c) (51 mg, 0.20 mmol) in methylene chloride (2 mL). The resulting mixture was stirred overnight at room temperature. After this time, HPLC indicated the end of the reaction. The reaction was then quenched by the addition of 1 N potassium sulfate. The mixture was then extracted with ethyl acetate. The extracts were combined, washed with brine, dried over magnesium sulfate and concentrated. The crude product obtained

The 163 product was purified by flash chromatography (3% methanol/methylene chloride) to give 101 mg of the desired product as a white foam. MS (M+H) ⁺ 792.4, (MH) ~ 790.7; IR (film) 1786 cm ^-1 , 1736 cm -1 , 1680 cm -1 , 1640 cm -1 .

e) (homochiral)

NH

H

COOH “VYJ

N—SO ₂ —N(CH ₃ ) ₂

A mixture of the product prepared in step (d) (95 mg, 0.12 mmol), 1 N HCl (120 μΐ, 0.12 mmol) and the catalyst, 10% palladium on carbon (49 mg) in dioxane was stirred under hydrogen atmosphere (hydrogen balloon) for 2 h at room temperature. After this time, analytical HPLC indicated the end of the reaction. The reaction mixture was then filtered through a pad of Celite ^R and concentrated to give the crude product (HCl salt). Purification by preparative HPLC (reverse phase, methanol, water, trifluoroacetic acid) followed by passage through a polyvinylpyrrolidone column gave 32 mg of the desired product as a white flaky powder.

MS 434.3 (M+H) ⁺ , 432.3 (MH)-; IR (KBr) 1778, 1663 cm-l.

Example 65 (homochiral)

_H2N

NH»HC1

O

O

II

O

II. c- N— CH ₂ — C— O—C (CH ₃ ) 3 ^k H o II

N—C—O—

164

and)

and?

(ch ₃ i ₃

A solution of the carboxylic acid derivative of the azetidinone prepared according to Example 1(b) (482 mg, 1.0 mmol) in tetrahydrofuran was cooled to -20°C under argon and N-methylmorpholine (223 mg, 2.2 mmol) was added. 1.1 equivalents of a 0.5 M solution of tert-butylglycine hydrochloride (184 mg, 1.1 mmol) were then added followed by benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (486 mg, 1.1 mmol). The reaction mixture was then stirred for 24 hours at -20°C, then poured into 5% potassium hydrogen sulfate solution and extracted with ethyl acetate. The ethyl acetate extract was washed with water and brine and the crude residue was purified by chromatography on silica gel using ethyl acetate as eluent to give 396 mg of the desired product as a colorless solid. MS 596 (M+H) ⁺ .

b) /δ\— ch ₂ — o- c- n ^x ~ ^/ o ¹¹ (ČP)-CHt-OC-

⁰

II H II

C- N— CH ₂ - C— O— C (CH ₃ ) 3

Y2/ ^N ~ J- O“ (CH ₂ ) ₂ -(O>

A solution of the product prepared in step (a) (200 mg, 0.336 mmol) and triethylamine (38 mg, 0.37 mmol) in methylene chloride (4 mL) was stirred at room temperature and then 1.1 equivalents of 1-phenethyloxypiperazine-4-carbonyl chloride (110 mg, 0.37 mmol) were added. Dimethylaminopyridine (10 mg) was then added and

165 • 9 9 I 959 9« • · · · · * · 9 · • · ♦ v · * » · · · • « « · · ···*»· · • t · · · · · · · · · · · · The reaction mixture was stirred for 30 hours. The reaction mixture was then diluted with methylene chloride, washed with brine, and dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography on silica gel using ethyl acetate to give 210 mg of the desired product as a colorless glassy residue. MS 856 (M+H) ⁺ .

c) (homochiral) nh.hci OO _> ^C_ n-ch ₂ -c— OC (CH ₃ ) ₃

Í~[ f~\ if r-, ς? ^γ ^Ν \_/ ^Ν ~ ^c “°- (CH ₂ ) ₂

O

A solution of the product prepared in step (b) (200 mg, 0.234 mmol) in dioxane (5 mL) containing 1.1 equivalents of HCl was stirred under nitrogen with the catalyst, 10% palladium on carbon (75 mg), for 2 h. The reaction mixture was then filtered and the solvents were evaporated to give 122 mg of the desired product as a colorless solid. MS 588 (M+H) ⁺ .

Example 66 (homochiral)

NH«HC1

H

\ “-CO— (CH ₂ > ₂

CN—CH ₂ —CO ₂ H

The product prepared according to Example 65 (30 mg, 0.05 mmol) was added to trifluoroacetic acid (1 ml) at 0 °C and the mixture was stirred for 30 minutes. The trifluoroacetic acid was then evaporated,

166 ·· ·· ·· • · » • ··* · · * · • · » · · · ··· • ^·

Example 67 (homochiral) r ^HC1 8 h _> ^c - ^{n ch} 3 ^H I /—\ 8

JN N— C— O— C— [CH(CH ₃ ) ₂ ]2 ° Y V_/ I o

and)

A solution of the carboxylic acid derivative of azetidinone prepared according to Example 1(b) (150 mg, 0.311 mmol) in tetrahydrofuran (3 ml) was cooled under argon to -20 °C and then N-methylmorpholine (34.6 mg, 0.342 mmol) was added. Then 1.1 equivalents of a 2 M solution of monomethylamine in tetrahydrofuran were added and then benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (151 mg, 0.341 mmol) was added. The reaction mixture was stirred for 48 hours at -20 °C, then poured into 5% potassium hydrogen sulfate solution and extracted with ethyl acetate. The ethyl acetate extract was then washed with water and brine, and dried over sodium sulfate. The solvents were then evaporated and the crude residue was purified by chromatography on silica using ethyl acetate to give 124 mg of the desired product as a colorless solid. MS 496 (M+H) ⁺ .

167

b) • · · · • · · • · · · • · · · • · · · 4 • · · · · ····· · · ·

A solution of the product prepared in step (a) (100 mg, 0.2 mmol) and triethylamine (23 mg, 0.225 mmol) in methylene chloride (2 mL) was stirred at room temperature and 1.1 equivalents of 1-diisopropylmethoxycarbonylpiperazine-4-carbonyl chloride (65 mg, 0.225 mmol) were added thereto. Dimethylaminopyridine (8 mg) was then added and the reaction mixture was stirred for 16 hours. The reaction mixture was then diluted with methylene chloride, washed with brine and dried over anhydrous sodium sulfate. Purification of the crude product by flash chromatography on silica using ethyl acetate gave 58 mg of the desired product as a glassy residue. MS 750 (M+H) ⁺ .

C)

ΝΗ.ΗΠ o (homochiral)

JI ¹¹ H

H ₂ N^ _aC-Ç-CH ₃

I £ /~Λ 1° ^C ~°“ ^{tCH ÍCH} 3 ^{} 2} ] 2

Oh H

A solution of the product prepared in step (b) (53 mg, 0.07 mmol) in dioxane (3 mL) containing 1.1 equivalents of 1 N HCl was stirred under hydrogen with a catalyst, 10% palladium on carbon (20 mg), for 2 h. The reaction mixture was then filtered and the solvent was evaporated to give 32 mg of the desired product as a colorless solid. MS 482 (M+H) ⁺ .

168 * ♦ · • · · · · • · · · ·

Example 68 (homochiral)

N—C—O—C (CH ₃ ) ₃

II about

about ₂ —ocn

C—N—CH ₃

Γ~\ /-N^ N N—O

J \_^ II o o

C—O—C(CH ₃ ) ₃

A solution of the product prepared according to Example 67(a) (120 mg, 0.242 mmol) and triethylamine (37 mg, 0.363 mmol) in methylene chloride (3.5 mL) was stirred at room temperature and 1.1 equivalents of 1-tert-butoxycarbonylpiperazine-4-carbonyl chloride (90 mg, 0.363 mmol) were added. Dimethylaminopyridine (6 mg) was then added and the reaction mixture was stirred for two hours. The reaction mixture was then diluted with methylene chloride, washed with brine and dried over anhydrous sodium sulfate. The crude product was purified by flash chromatography on silica using ethyl acetate as eluent to give 100 mg of the desired product as a colorless glass. MS 708 (M+H) ⁺ .

(homochiral)

N— C— O— C(CH ₃ ) ₃

169

• · · • · · · • » ·

A solution of the product prepared in step (a) (90 mg, 0.127 mmol) in dioxane (3 mL) containing 1.5 equivalents of HCl was stirred under hydrogen with a catalyst, 10% palladium on carbon (35 mg), for 2 hours. The reaction mixture was then filtered and the solvents were evaporated by lyophilization to give 38 mg of the desired product as a colorless solid. MS 439 (M+H) ⁺ ; [α]=14° (c=1, methanol).

Example 69

and)

O'

O (homochiral) c-nh ₂

N—C—O—C (CH ₃ ) 3 O

The desired product was prepared as a colorless solid by the method of Example 67(a) using 0.5 M ammonia in dioxane instead of 2 M monoethylamine. MS 482 (M+H) ⁺ .

b) (θ')— ch ₂ —o- c- n ⁰ II (θ^—CH7-0-C-

N

H

1?

170 • φ φφφφ

The desired product was prepared as a colorless glassy residue by reacting the product prepared in step (a) with 1-tert-butoxycarbonylpiperazine-4-carbonyl chloride according to the method of Example 68(a). MS 694 (M+H) ⁺ .

C)

NH.HC1 h ₂ n' ^A ' ^H ΧΪ ^N \_J - °- ^{c (CH} 3) ³ _k C-NH ₂ (homochiral)

The desired product was prepared as a colorless solid by deprotection and treatment of the product prepared in step (b) according to the method of Example 68(b). MS 426 (M+H) ⁺ .

Example 70 (homochiral)

The desired product was prepared as a colorless solid by the method of Example 67(a) using 1-tert-butoxycarbonylpiperazine instead of monoethylamine. MS • ·

171 • e · · ·

651 (M+H) ⁺ .

b)

A solution of the product prepared in step (a) (100 mg, 0.2 mmol) and triethylamine (23 mg, 0.225 mmol) in methylene chloride (2 mL) was stirred at room temperature and 1.1 equivalents of 1-diisopropylmethyloxycarbonylpiperazine-4-carbonyl chloride (65 mg, 0.225 mmol) were added. Dimethylaminopyridine (8 mg) was then added and the reaction mixture was stirred for 48 hours. The reaction mixture was then diluted with methylene chloride and dried over anhydrous sodium sulfate. Purification of the crude product by flash chromatography on silica using ethyl acetate as eluent gave 68 mg of the desired product as a colorless glass. MS 906 (M+H) ⁺ .

C)

NH«HC1 h ₂ n'^'n^ ^s * ^zZ45 'a H (homochiral)

O

C—O—C (CH ₃ ) ₃ /-=7 o

M ¹¹ \__y ^N ~ ^c ~°~ f “ ^[CH (CH ₃ ) ₂ ]:

A solution of the product prepared in step (b) (60 mg, 0.07 mmol) in dioxane (2 mL) containing 1.1 equivalents of 1.0 N HCl was stirred under hydrogen atmosphere with the catalyst, 10% palladium on carbon (25 mg), for 2 hours. The reaction mixture was then filtered and

» · ·

9

1 • 9 · 4

172 * · • · · · · · · · · lyophilization of the solvents yields 42 mg of the desired product as a colorless solid. MS 637 (M+H) ⁺ .

Example 71

To a solution of (R)-1-phenylaminopropane (420 mg, 2.82 mmol) and potassium carbonate (292 mg, 2.12 mmol) in tetrahydrofuran (5 mL) was added methyl iodide (200 mg, 1.41 mmol). The reaction mixture was stirred at room temperature for 4 h and then heated at 40-45 °C for 14 h. The reaction mixture was then filtered. The filtrate was concentrated and purified by flash chromatography [eluting with 5-10% ammonia (2 M in methanol) in methylene chloride] to give 43 mg of the desired product as a pale yellow oil.

To a solution of phosgene (0.265 ml, 0.536 mmol), 20% in toluene) in methylene chloride (1 ml) at 0 °C was added a solution of the product prepared in step (a) (40 mg, 0.268 mmol) and triethylamine (27 mg, 0.268 mmol) in methylene chloride (1 ml) over 10 minutes. The reaction mixture was then stirred for 1 hour at 0 °C. The reaction mixture was then

173 • · · · · · · · · · · · · · · · · · · · · ♦ • · · · · · ····· The mixture is concentrated and the residue is added to anhydrous ether (20 ml). The white solid formed is filtered off. The filtrate obtained is concentrated to give the desired product as a yellow oil.

A mixture of the benzyl ester product prepared according to Example 1(c) (80 mg, 0.14 mmol), the acid chloride derivative prepared in step (b) (44 mg, 0.21 mmol), dimethylaminopyridine (17.1 mg, 0.14 mmol) and triethylamine (21 mg, 0.21 mmol) in methylene chloride (3 mL) was stirred at room temperature for 7 hours. Purification of the reaction mixture by flash chromatography (using 25% ethyl acetate in hexane as eluent) gave 62 mg of the desired product as a colorless oil.

d)

A mixture of the product prepared in step (c) (60 mg, 0.08 mmol), catalyst, 10% palladium on carbon (8.48 mg, 0.0008

174

mmol), and 1 N HCl (0.08 mL, 0.08 mmol) in dioxane (3 mL) were stirred under hydrogen atmosphere (hydrogen balloon) for 1 h at room temperature. The reaction mixture was then filtered through a pad of Celite ^R . The filtrate was lyophilized to give 31 mg of the desired product as a white solid. MS 390.1 (M+H) ⁺ ; IR (film) 1780 caH.

Example 72 (homochiral)

NH«HC1

Η,ΐΛ _> ^C °2- ^CH 2- ^{CH (CH} 3 > 2 ^H Γ i?

N— C— N— C (CH ₃ ) ₃

O

a) <θ)-CH ₂ —OCN ⁰ /τt\ II Χχ , (Cý-CHy-OCN N ₄ CO ₂ -CH ₂ -CH(CH ₃ )2 'hi·.

NH

To a stirred solution of the azetidinone carboxylic acid prepared according to Example 1(b) (85 mg, 0.176 mmol) and 1-iodo-2-methylpropane (81 µL, 0.705 mmol) in dimethylformamide (500 µL) at room temperature was added cesium carbonate (29 mg, 0.088 mmol). After 24 hours, the reaction mixture was partitioned between ethyl acetate and water containing a small amount of sodium thiosulfate. The organic phase was separated, washed with saturated sodium chloride, dried over magnesium sulfate, and concentrated. The residue was purified by silica gel chromatography to give 62 mg of the desired product.

175 ··· * · · · · · · · · · · · · · · · · · · · · · · ····»· · ·

b)

O

The product prepared in step (a) (62 mg, 0.115 mmol) and the acid chloride product prepared according to Example 32 (c) were dissolved in methylene chloride (1.2 mL). Triethylamine (24 μΐ, 0.173 mmol) was then added followed by dimethylaminopyridine (3 mg, 0.023 mmol). After 12 hours, the reaction mixture was concentrated and the crude product was purified by silica gel chromatography to give 65 mg of the desired product.

c) (homochiral)

_H2N

NH«HC1

Λ

_to CO ₂ -CH ₂ -CH (CH ₃ ) ₂

YU about

N— C-NH •C(CH ₃ ) ₃

The product prepared in step (b) (65 mg, 0.087 mmol) was dissolved in 1,4-dioxane (1.0 mL). 1 N HCl (87 μΐ, 0.087) was then added followed by the catalyst, 10% palladium on carbon (12 mg). A hydrogen atmosphere was introduced via a balloon. After stirring at room temperature for 30 min, the reaction mixture was diluted with water:1,4-dioxane (1:1, 4 mL) and filtered. Lyophilization of the filtrate gave 47 mg of the desired product. IR (KBr) 1788 cm ^{- 1} .

176

Example 73 (homochiral)

NH»HC1

H _to CO ₂ -(CH ₂ ) ₂

^ΙΚ γ ^ζϊ \ν__7 ^Ν — C— N— C (CH3) 3

and)

The desired compound was prepared by the method of Example 72(a) using (2-iodoethyl)benzene instead of 1-iodo-2-methylpropane.

b)

"CO ₂ -(CH ₂ ) ₂

N— CN— C(CH ₃ ) ₃ H

The product prepared in step (a) (86 mg, 0.0147 mmol) and the acid chloride product prepared in Example 32(c) (51 mg, 0.206 mmol) were reacted according to the method of Example 72(b) to give 98 mg of the desired product.

177

c) (homochiral)

Deprotection and treatment of the product prepared in step (b) (97 mg, 0.122 mmol) according to the method of Example 72(c) gave 67 mg of the desired product. IR (KBr) 1790 cm ^-1 .

Example 74

_NH3CF3COOH

H ₂ N ^Λ N H

£O ₂ H (homochiral) /~Λ

VN N° _ΛΊ

C-O-CH—

The desired product was prepared as a colorless glassy mass by the method of Example 34, using cyclopentylmethanol instead of cyclohexylmethanol. MS 453.3 (M+H) ⁺ , 451.4 (MH) - ; IR (KBr) 1788 cm -1 , 1665 cm -1 .

Example 75 (homochiral)

NH«2HC1 u

H ₂ N Ν'

H /—\ í?

c—C—NH ₂ ~\ —\ ^K cA M )

II ' ' N—'

Oh H

178 • ·

a) ©—ano

II o—c—

V/ ¹ o

II

N—O-NH ₂

To a solution of N-carbobenzyloxypiperazine (5.5 g, 25 mmol) and diisopropylethylamine (9.6 mL, 7.1 gm, 55 mmol) in tetrahydrofuran (100 mL) at room temperature under argon was added dropwise over 15 min trimethylsilyl isocyanate (3.8 mL, 3.22 g, 28 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was then poured into water and extracted with ethyl acetate, washed with water and brine, and dried over sodium sulfate. The crude product was purified by column chromatography using 20% ethyl acetate/hexane to give 5.1 g of the desired product. MS (M+H) ⁺ 264.

b)

A mixture of the product prepared in step (a) (3.96 g, 15 mmol) and palladium on carbon catalyst (10%, 2 g) in methanol (100 mL) was stirred under hydrogen atmosphere (under a hydrogen balloon) for 1.25 h at room temperature. Filtration and concentration of the reaction mixture gave 2.0 g of the desired product.

H

About /'

-NH

179

A mixture of the azetidinone product in the carboxylic acid form prepared by the method of Example 1(b) (180 mg,

0.37 mmol), the product prepared in step (b) (62 mg,

0.48 mmol), diisopropylethylamine (84 μΐ, 0.48 mmol), ethyl-3-(dimethylamino)propylcarbodiimide hydrochloride salt (92 mg, 0.48 mmol) and 1-hydroxy-7-azabenzotriazole (65 mg, 0.48 mmol) in tetrahydrofuran (10 ml) were heated overnight at 60 °C. The reaction mixture was then diluted with methylene chloride, washed with water, dried over magnesium sulfate and concentrated to give the crude product. Purification of the crude product by flash chromatography (silica, 5-10% methanol/methylene chloride) gave 128 mg of the desired product as a white solid. MS 594.3 (M+H) ⁺ , 592.3 (MH) ^- .

A mixture of the product prepared in step (c) (110 mg, 0.185 mmol), the acid chloride prepared according to the method of Example 56(a) (63 mg, 0.28 mmol), diisopropylethylamine (96 µL) and 4-dimethylaminopyridine (18 mg) in methylene chloride (4 mL) was stirred for 18 h at room temperature. The reaction mixture was then quenched with saturated sodium chloride solution and extracted with ethyl acetate (3 x 50 mL). The extracts were combined, dried over magnesium sulfate and concentrated. The crude product was purified by flash chromatography (0-15% methanol/methylene chloride) to give 114 mg of the desired product as a white solid. MS 784.5 (M+H)+, 782.5 (MH) ^- ; IR (KBr) 1782 cm-1,

180

1732 cm ^-1 , 1643 cm ^-1 , 1586 cm ^-1

E)

_H2N

NH.2HC1 rf ₀ (homochiral) ir\ ř

C—1'^ N— c-nh ₂ ' N—v

N—< Λ /O C

--- N

About H <N

A mixture of the product prepared in step (d) (1.39 g, 1.77 mmol), 1 N HCl (3.54 mL, 3.54 mmol) palladium on carbon catalyst (10%, 750 mg) in dioxane (30 mL) was stirred under hydrogen atmosphere (under a hydrogen balloon) at room temperature for 3.5 hours. After this time, analytical HPLC indicated the end of the reaction. The reaction mixture was then filtered through a pad of Celite ^R and lyophilized to give 1.01 of a white solid. MS 260.6 (M+2H)2+; IR 1780 cmi, 1632 cmi.

NH χ

H ₂ NN' H

IV:

Other compounds of the general formula _K Ri have also been prepared

-N

X2 example

X2

R1 salt stereochemistry (M+H) ⁺

Oh,

II / ^C -\J

NCN—C(CH ₃ ) ₃

1.0 HCl homochiral 536

I? /-\ H “ ^C ”\ /~ ^C-NH 2

181

I · · « • · · « « • · · example

_X2

R1 salt stereochemistry (M+H) ⁺

About _{3 3}

O

II ch ₃

-COzH homochiral 453

-cn-(ch ₂ ) ₂ -n— cch ₃

CH ₃ •ch ₃

_-CO2H

About ch ₃ ch ₃ θ

-CN-(CH ₂ ) ₂ -N—C—O-ťpH—CH(CH ₃ )2 CH(CH ₃ ) ₂ homochiral 471

About r~\ »

1.0 HCl homochiral 539 _£_ _n ' jq— _C —O-CH—CH(CH ₃ ) ₂ 'VA

CH(CH ₃ ) ₂ oo

II II

-CN-(CH ₂ ) ₂ -C-NH ₂ ° _ O 2.0 CF 3 COOH

CH(CH ₃ ) ₂

595 homochiral

- C-NH-(CH ₂ ) ₂ “ N- [CH (CH ₃ ) ₂ ] ₂

OO 1.0 HCl homochiral 579 _ L C- O- CH- CH (CH ₃ ) 2 \_y I

CH(CH ₃ ) ₂

II / \ 11

-C-N )— C—

_NH2

182 • · ·· · · · · · • · · · · · · · ··· · ··· · ··· · « · · · · · ········

example

_X2

R1 salt stereochemistry (M+H) ⁺

1.0 HCl homochiral 615 '- ¹ CH(CH ₃ ) ₂

O

II

C—N. C—NH ₂

H II

O

O _ ysí— C- O- CH— CH (CH ₃ ) ₂

1.0 CF3CO2H 627 homochiral

CH(CH ₃ ) ₂

- C- - CH ₂ —©

O ₀ _ — c— O- CH- CH (CH ₃ ) ₂ \_7 I

CH(CH ₃ ) ₂

2.0 CF3CO2H 537 homochiral

O

II /"\

-C-N NH 'V?

OO ¹¹ '““Vr-o_C—NN ^{c 0} 'V/

CH-CH(CH ₃ ) ₂

CH(CH ₃ )2

1.0 CF3CO2H 626 homochiral

183 ··· · » Φ 9 9 9 ··<>* · ·«· 9 9 • · ·«· 9 9 9 9 9 9 9 · example X2 Ri salt stereochemistry (M+H) ⁺

-cn-ch ₂ —c-nh ₂ H

88 0 II —c- /—\ •\_y -CO2H --- homochiral 616 N- C- 0- CH— CH(CH ₃ ) ₂ 0 CH(CH ₃ ) ₂ 89 0 -CO2H --- homochiral 426 II /—What? -c— 1^^t-CN(C ₂ H 5 ) 2

184 example

X2

Z"λ "C—NN— CN-CH-CH(CH ₃ ) •\_y'

CH(CH ₃ ) ₂

O

II — c— _ o

N— c- N [ CH (CH ₃ ) • 0 · · 0 0 0 • · · 0 0 0

0 0 0 · 0*0

0 000 0000

0 0 0 00 00 0

Ri salt stereochemistry (M+H) ⁺

-CO2H --- homochiral 468

2)2

O _t __ O __ O

-CJ^_^N'C-1^_N- COC (CH ₃ )

O

II r~\ — c—N N-C-o·

-CH-O

-CO2H --- homochiral 454

-CO2H --- homochiral 539

-CO2H homochiral 549

1.0 HCl

-CO2H homochiral

1.0 CF3CO2H

O

- ^C -\J _N - c- O-CH- CH ₂ - CH (CH ₃ ) 2

_CH2 -CH( _CH3 ) ₂

497

185 ·· 9 9 999 99 » · « 9 9 9 9 9

9999 9 9»9 · « • · 999 9999999 9

9999 9» 99 « 99 »99 example X2

Ri salt stereochemistry (M+H) ⁺

-CO2H --- homochiral 466

-CO2H --homochiral 540 — C-

_CH2 -CH( _CH3 ) ₂

NCOC-(CH ₃ ) ₃

O

-CO2H --homochiral 540

O

II •cVy ^{N c} ^ .CH ₂ -CH(CH ₃ ) 2

N—C—OC—(CH ₃ ) ₃ Η ||

O

-CO2H --- homochiral 616 » π . _/p\

-c-^ _N -c _CH2 -^O>

N - C- O- CH— CH (CH ₃ ) ₂ O CH(CH ₃ ) ₂

186 ·· ·· «· · #· • · · · · · · * · « · · · · ··· · · • » ft·· ·····»· · ···« ΛΛ «· · ·· ···

example X2 Ri salt stereochemistry (M+H) ⁺ 99 -c-v7 ^c- O -CO ₂ H 1.0 HCl homochiral 437 100 í? /—v 2 /ΤΛ -CO2H 1.0 HCl homochiral 431

101 ⁰ η

II /—\ ír — C-Ν Ν—C—Ο\__y

-CO2H homochiral

0.02 CF3CO2H

469

CH— CH(CH ₃ )2 CH(CH ₃ ) ₂

102

CH ₃

-ch ₃

-CO2H homochiral 451

1.0 CF ₃ CO ₂ H íí _{__} O /\ II c- u— c— CH- CH ( ch ₃ ) 2

CH(CH ₃ ) ₂

103

-CO ₂ H --- homochiral 453

187 • · • · • c example

X2

Ri salt stereochemistry (M+H) ⁺

104 fř r-Ν N \_y

H

2.0 HCl homochiral 519 íř r~'\ π

V-C-NH2 ir

105

2.0 CF3CO2H

106

107

O

II — what

II

N-Cθ'

CH-CH(CH ₃ ) ₂

AND

CH(CH ₃ )2

homochiral 649 íi

-c-CO2H homochiral 341

1.0 HCl

-CO2H --homochiral 441 • · · · · · · * example

X2

188

R1 salt stereochemistry (M+H) ⁺

O

II r~\ — _N— SO2-CH2- CH(CH ₃ ) 2

110 ch _{3 o} ch ₃ II II II

CN-(CH ₂ ) 2“N—C o CH ₃

1.0 HCl homochiral 563

CH ₃

CH ₃ ch ₃ nh ₂ ⁰ /—\

-CH/— ^C °2 ^H

O—CH ₃

O

O—CH ₃

-CO2H homochiral

1.0 HCl

370

-CO2H --- homochiral 512

112

189 example X ₂ Ri salt stereochemistry (M+H) ⁺

113 -CO2H --- homochiral 512

O—CH ₃

I o o

II /—\ II — C— ^N-C—

O—CH ₃

114

-CO2H homochiral 468

About About

C-kQVČ— CH ₂ -N[CH(CH ₃ ) ₂ ] ₂

2.0 CF3CO2H

115

-CO ₂ H --- homochiral 496

1?

C-I^_N

About About

C— C— N— CH— CH (CH ₃ ) Η I

CH(CH ₃ ) ₂

190 example X2

Ri salt stereochemistry (M+H) ⁺

118

O

-CVJ

NC

-CO2H homochiral 434

2.0 CF3CO2H

119

-Ch ₃ c nh ₂ /-Λ n

-CO2H homochiral 439

2.0 CF3CO2H

120

O — cJ \jC-CH ₂ -N-}CH(CH ₃ ) ₂

V-/ II

2.0 CF3CO2H 579 homochiral

121 ? /~\ cVy'

N—C—NH ₂

-CO2H --- homochiral 512 íř /—\ íl

-CN N—C—C—CH ₃ '- ^/ H ₃ C N-jf—O—C(CH ₃ ) ₃

O

AC about

II

NCOC(CH ₃ ) ₃

H

122

-CO2H --- homochiral 427 • ·

191 example X2 Ri salt stereochemistry (M+H) ⁺

123 -CO2H mixture of homo- 461

125 θ 1.0 HCl homochiral 494 _ !L /A— C- O— C (CH ₃ ) 3 \_?

-it

126 o

v _ c— N- C— O- ChJ CH (CH3) 2

2.0 CF3COOH homochiral

551

127 O

-C-

NC— O-CH- CH(CH ₃ ) ₂

2.0 CF3COOH homochiral

594

192

example

Xa

Ri salt stereochemistry (M+H) ⁺

128

2.0 CF3COOH 619 homochiral

129

2.0 CF3COOH

-CO ₂ H homochiral 567

130 ff

2.0 CF3COOH 594 homochiral “ C- N- C— O- CH-| CH (CH ₃ ) íí / \ í?

- c—1^_M - CH ₂ - c—nh ₂

Other compounds of the general formula were also prepared

IV;

example

X2

193

131 íř z-Λ ř

-C—Ιξ_^N— c— NC(CH ₃ ) ₃

132 íř /—p.

•C—_N-

133 ir

134

About 11

-c-\J

Rl salt stereochemistry (M+H) ⁺ homochiral 426

2.0 HCl homochiral 520

C“NN—C—NH ₂

-CO2H homochiral 409

1.0 HCl

1.0 HCl homochiral 579

N-C-O-CH

CH(CH ₃ )2

CH(CH ₃ ) ₂

íř c—nh ₂

194 (racemate)

Example 135

_H2N

NH«HC1

--CH ₃

JN ⁰ SÍ(CH ₃ ) ₂

C(CH ₃ ) ₃

4,4-Dimethyl-2-azetidinone (17 g, 0.171 mol) and tert-butyldimethylsilyl chloride (28.43 g, 0.188 mol) were dissolved in methylene chloride (270 ml). A solution of diisopropylethylamine (44.80 ml, 0.257 mol) in methylene chloride (130 ml) was then added dropwise. The reaction mixture was then stirred for 24 hours at room temperature and then concentrated. The residue was partitioned between ethyl acetate and water. The organic phase was washed with 1 N potassium hydrogen sulfate, saturated sodium carbonate, dried over magnesium sulfate and concentrated. Purification of the residue by chromatography on silica gel gave 34.55 g of the desired product.

To a stirred solution of isopropylamine (1.08 mL, 7.73 mmol) in

195 • 9

9

9··· · · ♦ ·

« · 9 · tetrahydrofuran (5.0 ml) was added dropwise to a 1.6 M solution of butyllithium in hexane (4.83 ml, 7.73 mmol) at 0 °C. After 30 minutes, the solution was cooled to -78 °C and a solution of the product prepared in step (a) (1.50 g, 7.03 mmol) in tetrahydrofuran (2.5 ml) was added dropwise. After 40 minutes, a solution of 1-chloro-3-iodopropane (1.72 g, 8.43 mmol) in tetrahydrofuran (2.5 ml) was added dropwise. The temperature was then slowly increased to 0 °C. After 1 hour, the reaction was quenched by the addition of 1 N potassium hydrogen sulfate. The solution was then partitioned between ethyl acetate and water. The organic phase was washed with 1 N potassium hydrogen sulfate and saturated sodium chloride, dried over magnesium sulfate and concentrated. Purification of the residue by chromatography on silica gel afforded 1.46 g of the desired product.

C)

C(CH ₃ ) ₃

To a stirred solution of the product prepared in step (b) (1.36 g, 4.69 mmol) and tetrabutylammonium iodide (0.35 g, 0.94 mmol) in dimethylformamide (10 mL) was added sodium azide (0.91 g, 14.07 mmol). The reaction mixture was heated at 45 °C for 9 hours, then the solution was cooled to room temperature and partitioned between ethyl acetate and water. The organic phase was washed with saturated sodium chloride, dried over magnesium sulfate and concentrated to give 1.34 g of the desired product.

d) h ₃ ccooh»h ₂ n

C( _CH3 )3

196

The product prepared in step (c) (0.57 g, 1.92 mmol) was dissolved in 1,4-dioxane. Acetic acid (0.11 mL, 1.92 mmol) was then added followed by the catalyst, 10% palladium on carbon (0.15 mol %). A hydrogen atmosphere was introduced via a balloon. After stirring at room temperature for one hour, the solution was filtered and concentrated to give 0.59 g of the desired product.

E)

C(CH ₃ ) ₃

The product prepared in step (d) (0.59 g, 1.78 mmol) was dissolved in acetonitrile (8.0 mL). Triethylamine (0.26 mL, 1.87 mmol) was then added followed by N,N'-dicarbobenzyloxy-S-methylisothiourea (0.64 g, 1.78 mmol). After stirring at room temperature for 12 hours, the solution was partitioned between ethyl acetate and water. The organic phase was washed with saturated sodium chloride, dried over magnesium sulfate and concentrated. Purification of the residue by chromatography on silica gel gave 0.37 g of the desired product.

F)

197

To a stirred solution of the product prepared in step (e) (0.36 g, 0.62 mmol) in tetrahydrofuran (5 mL) at 0 °C was added dropwise a 1.0 M solution of tetrabutylammonium fluoride in tetrahydrofuran (0.69 mL, 0.69 mmol). The reaction mixture was then stirred at room temperature. After 1 hour, the solution was partitioned between ethyl acetate and water. The organic phase was washed with saturated sodium chloride, dried over magnesium sulfate and concentrated. Purification of the residue by chromatography on silica gel gave 276 mg of the desired product.

G)

Oh H

To a stirred solution of the product prepared in step (f) (0.12 g, 0.26 mmol) in tetrahydrofuran (1.5 ml) at -78 °C was added dropwise a 1.0 M solution of bis(trimethylsilyl)amide in tetrahydrofuran (0.28 ml, 0.28 mmol). After stirring for 30 minutes, phenylisocyanate (42 μΐ, 0.39 mmol) was added dropwise. The temperature was then slowly raised to 0 °C. After 30 minutes, the reaction was quenched by addition of a pH 4.0 buffer solution. The resulting mixture was partitioned between ethyl acetate and water. The organic phase was washed with saturated sodium chloride, dried over magnesium sulfate and concentrated. Purification of the residue by chromatography on silica gel gave 0.16 g of the desired product.

• · • ·

198 • · · · • · · »· · · * (racemate)

h)

NH«HC1 u

H ₂ N N'

H

CH ₃ •ch ₃

The product prepared in step (g) (0.16 g, 0.27 mmol) was dissolved in 1,4-dioxane. Then, a catalyst, 10% palladium on carbon (0.15 mol %) was added, followed by 4 N HCl in 1,4-dioxane (68 μΐ, 0.27 mmol). A hydrogen atmosphere was introduced via a balloon. Water (0.20 mL) was added to obtain the product. After another 30 minutes, the reaction mixture was diluted

50% acetonitrile in water (2.0 mL) and filtered. The solution was then concentrated to remove organic solvents and lyophilized to give 84 mg of the desired product. IR (KBr) 1753, 1707 cm-1; (M+H) ⁺ 318.

did not fall out of solution after minutes of mixing.

Other compounds of general formula II were also prepared:

N ^e H .0

199 • · • · · • · «

example Xi salt stereochemistry (M+H) ⁺ 136 0 II — c-ch ₃ 1.0 cf ₃ co ₂ h racemate 241 137 1.0 HCl racemate 303

138 1.0 HCI racemate 379 139 1.0 HCI racemate 371

140 0 — C-CHs-O— 1.0 HCl racemate 333

200 • · • ·

example Xi salt stereochemistry (M+H) ⁺ 141 1.0 HCl racemate 382 0 _ — C-NH-SO ₂ -^ý 142 1.0 HCl racemate 332 ch ₃ 143 1.0 CF3CO2H racemate 336 0 — C-NH— 144 1.0 HCl racemate 395 -so ₂ -@-c (CH ₃ ) 3 145 1.0 CF3CO2H racemate 339 -sat ₂ -@

201 • ·

Example 146 (homochiral)

The racemic product prepared according to Example 135 step (f) was separated on a Chiralpak-AD preparative column using 30% 2-propanol/hexane as eluent into pure enantiomeric (-) and (+) isomers.

A solution of the (+) isomer prepared in step (a) (125 mg, 0.268 mmol) in tetrahydrofuran (2 ml) was cooled to -78 °C under argon. A 1 M solution of bis(trimethylsilyl)amide (0.536 ml) in tetrahydrofuran was then added dropwise and the mixture was stirred for 20 minutes. Benzenesulfonyl chloride (95 mg, 0.536 mmol) was then added dropwise and the mixture was stirred for 4 hours.

· · · » 9 9··

202 » · · • ř · · hours at -78 °C. The reaction mixture was then diluted with 10% aqueous potassium hydrogen sulfate (10 mL) and then extracted with ethyl acetate (3 x 10 mL). The combined organics were washed with water (25 mL) and brine (25 mL), and dried over sodium sulfate. The solution was then filtered and the solvent was evaporated to give an oil. Purification of the residue by flash chromatography (silica, ethyl acetate:hexane 1:3) gave 149 mg of the desired product as a colorless oil. (M+H) ⁺ 607.

c) (homochiral)

_H2N

NH4OHCl

A solution of the product prepared in step (b) (140 mg, 0.23 mmol) in dioxane (4 mL) containing aqueous 1 N HCl (0.46 mL) and the catalyst, 10% palladium on carbon (70 mg), was stirred for 1 h under hydrogen. The reaction mixture was then filtered and lyophilized to give 78 mg of the desired product as a colorless solid. IR (KBr) 1778 cm 1; (M+H) ⁺ 339; [α]β = +12° (methanol), c = 1).

Other compounds of the general formula have also been prepared

NH h ₂ n^ n^^ ^//ó ·

H

CH ₃ ch ₃

Xi

II:

203 ·· ·· ► · · • < ♦· • · « · « »·»· ·· example

Xi salt stereochemistry (M+H) ⁺

147

1.0 CF3CO2H homochiral 409

- c— ch ₂ - o—

148

1.0 HCl homochiral 415

1.0 HCl homochiral 450

150

1.0 HCl homochiral 333

151

1.0 HCl homochiral 349

O

II

-o ch ₂ -o·

QVoh

204 • 9 • ··

example Xi salt stereochemistry (M+H) ⁺ 152 _o - c— ch ₂ - 0— 1.0 HCl homochiral 383 153 c— ch ₂ - 0 Q-^O^ 1.0 HCl homochiral 425 154 — homochiral 353

155

156

AND?

and?

—C-CH2-O-CH3

1.0 HCl homochiral 270

1.0 HCl homochiral 359 —c— ch ₂ - o— (CH ₂ ) ₂ —O— (CH ₂ ) 2“ O ^- ch ₃ íř

157

1.0 HCl homochiral 409

205 • · · · • · · • · · · • 9 · · • · · e · · · · ·

example Xi salt stereochemistry (M+H) ⁺ 158 1.0 CF3CO2H homochiral 409 0 -c— ch ₂ —o- YES) (Ό) 159 1.0 HCl homochiral 466 0 -ř- Sat ₂ ~N(C ₃ H ₇ ) ₂

Example 160 (homochiral)

and)

• · • ·

206

9 9 9 9 99*

According to Example 146, step (a), the prepared (-) isomer (0.163, 0.349 mmol) was dissolved in tetrahydrofuran (2 ml) and cooled to -78 °C. Sodium bis(trimethylsilyl)amide (0.52 ml, 0.524 mmol) was then added dropwise. The resulting mixture was stirred for 20 minutes. Benzenesulfonyl chloride (93 mg, 0.524 mmol) was then added and the reaction mixture was stirred for 1.5 hours at -78 °C and then stirred overnight at room temperature. The reaction was then quenched by the addition of 0.5 N potassium hydrogen sulfate solution (25 ml) and extracted with ethyl acetate (2 x 20 ml). The organic phase was then washed with brine (1 x 40 ml) and filtered over sodium sulfate. The filtrate was evaporated to a colorless oil. Purification of this oil by preparative HPLC (YMC ODS A 30 x 250 mm, 5 μ column) gave 108 mg of the desired product as an oil. (M+H) ⁺ 607.2; [α]ϋ=-7.0 ^θ (c=0.3, chloroform).

To a solution of the product prepared in step (a) (105 mg, 0.173 mmol) in 1,4-dioxane (15 mL) containing 1 N HCl (0.21 mL) was added the catalyst, 10% palladium on carbon (50 mg). Hydrogen was then bubbled through the solution for 1 hour. The reaction mixture was then filtered through a pad of Celite ^R and washed again.

1,4-dioxane. The filtrate and washings were combined and evaporated to give a colorless oil (53 mg). This oil was then further purified by reverse phase preparative HPLC (YMC ODS A 20 x

207

100 mm, 5 μ, fast elution column) and 23 mg of the desired product was obtained as an oil. (M+H) ⁺ 339; [α]ϋ=-9.3 ^θ (c=0.42, methanol).

Another compound of general formula II was also prepared:

_H2N

NH

H

_CH3 — _CH3

example Xi salt stereochemistry (M+H) ⁺ 161 —c-ch ₂ -o~^5) 1.0 CF ₃ CO2H homochiral 333

Other compounds of general formula II were also prepared:

208 ··· · · · · · • · · * · ft · · · « • · ··« ······· · ······ ·« 4 «·

example Xi Ri Ru salt stereochemistry (M+H) ⁺ 162 — C-NH— -every ₂ hours CH ₃ racemate 348 163 -Lh-@ -CO2CH3 ch ₃ 1.0 HCl racemate 362

Other compounds of general formula II were also prepared

NH

X h ₂ n N -j ^- *

Rn

LftRi

N

Xi

example Xi Ri Ru salt stereochemistry (M+H) ⁺ 164 -CO2H C-NH— CH ₃ 1.0 CF ₃ CO ₂ H racemate 348 165 -CO ₂ CH ₃ -C-NH-^^ ch ₃ 1.0 HCl racemate 362

209

Example 166

O

NH· 1.0 HCl ||

Λ. £ ^H 3 ^C-OCHa (racemate)

_H2N

N

N

The above racemic compound is prepared from D,L-aspartic acid by the method for preparing a chiral compound from L-aspartic acid (P.E. Finke et al.,

J.Med.Chem., Vol.38, p.2449, 1995).

To diisopropylamine (5.6 mL, 40 mL) in tetrahydrofuran (30 mL) was added 2.5 M butyllithium (14 mL) in hexane (35 mmol) under argon at -20 °C. The resulting mixture was stirred for 15 min and then cooled to -70 °C. A solution of the racemic product prepared in step (a) (4.00 g, 17.4 mmol) in tetrahydrofuran (16 mL) was then added over 5 min and the reaction mixture was warmed to -20 °C over 15 min. A solution of methyl iodide (2.72 mL, 43.7 mmol) in tetrahydrofuran (4 mL) was then added and the reaction mixture was stirred for 30 min at a temperature between -20 °C and 0 °C. • · • >

210 with dry ethyl ether (50 mL) and the reaction mixture poured into a mixture of ice and 80 mL of 1N HCl. The layers were then separated and the aqueous layer was extracted twice with brine, dried over sodium sulfate and concentrated to an amorphous solid. Workup with hexane and ethyl acetate gave 2.86 g of the desired product as a crystalline mass.

C)

Cl·

Si(CH ₃ ) ₂

C(CH ₃ ) ₃

To diisopropylamine (3.62 mL, 25.8 mmol) in tetrahydrofuran (20 mL) was added butyl lithium (9.2 mL) in hexane (23 mmol) under argon at -20°C. The resulting mixture was stirred for 15 min and then cooled to -70°C. A solution of the product prepared in step (b) (2.76 g, 11.3 mmol) in tetrahydrofuran (12 mL) was then added over 5 min and the reaction mixture was then warmed to -20°C over 15 min. A solution of 1-chloro-3-iodopropane (2.5 mL, 23.3 mmol) in tetrahydrofuran (5 mL) was then added and the reaction mixture was stirred for 30 min at a temperature ranging from -20°C to 0°C. Dry ethyl ether (50 mL) was then added and the reaction mixture was poured into a mixture of ice and 52 mL of 1N HCl. The layers were separated and the aqueous layer was extracted twice with ethyl ether. The combined extracts were washed twice with brine, dried over sodium sulfate, and concentrated to an oil. Repeated concentration of the oil from ethyl ether and hexane afforded 3.35 g of the desired product as an oil.

d)

CH ₃

II

Cl·

Si(CH ₃ ) ₂

C(CH ₃ ) ₃ o

211 ··· ·«· ··« «*··· b «·« · · • · ··« ······» · ·«· · · · · · «·«·«· ·· · · · ·

The product prepared in step (c) (1.92 g, 6 mmol) in ethyl ether (30 ml) was treated with excess diazomethane in ether in an ice-water bath until a permanent yellow color was obtained. The mixture was then flushed with nitrogen for 10 minutes and the solution was concentrated. The oily residue was taken up in ethyl ether and the solution was washed with cold dilute potassium hydrogen sulfate and then brine (twice), dried over sodium sulfate and concentrated to give 2.046 g of the desired product as an oil.

A mixture of the product prepared in step (d) (2.95 g, 8.83 mmol), sodium azide (2.44 g, 35.3 mmol) and tetrabutylammonium iodide (2.45 g, 6.64 mmol) in dimethylformamide (12 mL) was stirred under argon at 60°C for 16 h. This reaction mixture was then combined with a second reaction mixture obtained with the product from step (d) (334 mg, 1 mmol). The dimethylformamide was then removed in vacuo and the residue was taken up in ethyl acetate and diluted with aqueous lithium chloride. The ethyl acetate layer was then washed again with dilute lithium chloride and then brine (twice) and concentrated to give 3.53 g of the desired product as a crude viscous oil.

• ·

212

A solution of the product prepared in step (e) (3.53 g) in tetrahydrofuran (30 mL), acetic acid (1.2 mL) and 1.0 M tetrabutylammonium fluoride (20 mL) in tetrahydrofuran was stirred at room temperature for 2 hours, then concentrated to a residue which was taken up in ethyl acetate and brine. After extraction, the ethyl acetate layer was washed with brine (twice), dried over sodium sulfate and concentrated to an oil (9.33 g). Chromatography of the resulting oil using ethyl acetate:hexanes (7:3) gave 2.1 g of the desired product as an oil.

G)

HC1«H ₂ N' ch ₃ ,c-och ₃

The product prepared in step (f) (678 mg, 3 mmol) was hydrogenated for 2 hours in 20 mL of dioxane and 3.0 mL of aqueous 1 M HCl in the presence of a catalyst, 10% palladium on carbon (237 mg). The mixture was then filtered using aqueous dioxane, the filtrate was concentrated, and the residue was reconcentrated with dioxane to give 974 mg of the desired product as a crude thick gum.

213 • · ··«··· · · · · · ·

h)

To a solution of the product prepared in step (g) (945 mg) in dry methanol (7 ml) and dry dimethyl ether (7 ml) under argon atmosphere, N,N'-dicarbobenzyloxy-S-methylisothiourea (1.61, 4.5 mmol), triethylamine (1.5 ml,

10.7 mmol) and mercuric chloride (1.22 g, 4.5 mmol). The reaction mixture was stirred for 2 hours at room temperature and then filtered through Celite ^R with ethyl acetate. The filtrate was concentrated to give a residue which was taken up in ethyl acetate and diluted with potassium bisulfate. Extraction was then carried out twice with ethyl acetate, the combined ethyl acetate extracts were washed with brine (twice), dried over sodium sulfate, and concentrated to an oil (2.25 g). Purification by chromatography on silica gel (150 g) using ethyl acetate:hexane (6:4) gave 1.03 g of the desired product as an oily residue.

and)

N

H

To a solution of the product prepared in step (h) (582 mg, • ·

214

1.14 mmol), previously dried by azeotropic evaporation with tetrahydrofuran and toluene in tetrahydrofuran (8 ml), was added 1.0 M sodium bis(trimethylsilyl)amide (1.5 ml, 1.5 mmol) at -78 °C. The resulting mixture was stirred for 15 min and then phenylisocyanate (150 μΐ, 1.38 mmol) was added. The reaction mixture was then warmed to 0 °C over 30 min and poured into 8 ml of 10% potassium hydrogen sulfate in water. After extraction with ethyl acetate (three times), the combined ethyl acetate extract was washed with water (twice), dried over sodium sulfate, and concentrated to give a viscous oil (0.78 g). Purification by chromatography (silica gel, 60 g) using ethyl acetate:hexanes (35:65) gave 656 mg of the desired product as an oily residue.

3) (racemate)

O

NH· 1.0 HCl ||

She £ ^H 3 aP-och ₃

_H2NNH

-N.

The product prepared in step (i) (636 mg, 1.01 mmol) was hydrogenated for 2 hours in 12 ml of dioxane and 1.0 ml of 1 N HCl (1 mmol) in the presence of a catalyst, 10% palladium on carbon (223 mg) at a pressure of 1 atmosphere. After filtration using dioxane, the filtrate was concentrated and lyophilization of the residue from aqueous acetonitrile gave 336 mg of the desired product as a hygroscopic solid. IR (KBr) 1775 cm-1; (M+H) ⁺ 362.

• ·

215

Another compound of the general formula was also prepared

III:

example Xi

Ri

R12 salt stereochemistry (M+H) ⁺

167 CH3 1.0 HCl racemate 415

O

216 ···· ·· ·· · ·· ·· ··

To a solution of diisopropylamine (8.9 mL, 63.5 mmol) in tetrahydrofuran (50 mL) at -20°C under argon was added 22.6 mL of 2.5 M butyllithium hexane (56.6 mmol). The mixture was stirred for 15 minutes and then cooled to -70°C. A solution of the racemic compound prepared according to Example 166, step (a) (6.49 g, 28.3 mmol) in tetrahydrofuran (30 mL) was then added over 5 minutes and the reaction mixture was warmed to -20°C over 15 minutes. A solution of 1-chloro-3-iodopropane (6.3 mL, 58.7 mmol) in tetrahydrofuran (10 mL) was then added and the reaction mixture was stirred for 30 minutes at a temperature between -20°C and 0°C. Dry ethyl ether (125 mL) was added and the reaction mixture was poured into a mixture of ice, 1 N HCl (130 mL) and ethyl ether. After separation of the layers, the aqueous aqueous layer was extracted with ethyl ether (twice). The combined ethyl ether extract was washed with brine (twice), dried over sodium sulfate, and concentrated to an oil which upon further concentration from hexanes (5 x) gave 7.61 g of the desired product as an oil.

217

To diisopropylamine (4.7 mL, 33.5 mmol) in tetrahydrofuran (25 mL) at -20°C under argon was added 12 mL of 2.5 M butyllithium in hexane (30 mmol). The mixture was stirred for 15 min and then cooled to -70°C. A solution of the product prepared in step (a) (4.50 g, 14.7 mmol), previously dried by azeotropic evaporation from toluene and tetrahydrofuran, in tetrahydrofuran (20 mL) was then added over a period of 15 min and the reaction mixture was warmed to -20°C over a period of 15 min. A solution of methyl iodide (1.90 mL, 30.5 mmol) in tetrahydrofuran (6 mL) was then added and the reaction mixture was stirred for 30 min at a temperature between -20°C and 0°C. Dry ethyl ether (70 mL) was added and the reaction mixture was poured into a mixture of ice, 1 N HCl (66 mL) and ethyl ether.

After separation of the layers, the aqueous aqueous layer was extracted with ethyl ether (twice). The extracts were combined, washed with brine (twice), dried over sodium sulfate, and concentrated to an oil which upon further concentration from hexanes (5x) gave 4.60 g of an oil containing the 3α-methyl epimer and the corresponding 3β-methyl epimer in a 4:1 ratio.

3α-methyl-epimer + 3β-methyl-epimer

Treatment of the mixed product prepared in step (h) (1.24 g) with diazomethane according to the method of Example 166, step (d) gave 1.36 g of an oil containing the 3α-methyl epimer and

218 • · · · • · · · · · · corresponding 33-methyl-epimer in a ratio of 4:1.

3α-methyl-epimer + 3β-methyl-epimer

Treatment of the mixed product prepared in step (c) (1.35 g) with sodium azide and tetrabutylammonium iodide in dimethylformamide according to the method of Example 166, step (e) gave 1.21 of a crude oil containing 3α-methyl-epimer and 3β-methyl-epimer.

E)

About ^Sli3 i'

J-NH

O

3α-methyl-epimer + 33-methyl-epimer

Treatment of the crude mixed product prepared in step (d) (1.21 g) in tetrahydrofuran (10 ml) with 5.0 ml of 1.0 M tetrabutylammonium fluoride in tetrahydrofuran and acetic acid (0.3 ml) according to the method of Example 166, step (f) gave, after chromatography on 100 g of silica gel using ethyl acetate:hexanes (1:1), 343 mg of an oil containing the 3α-methyl epimer and the corresponding 3β-methyl epimer in a ratio of (86:14). Chromatography then gave an additional 256 mg of the mixture.

• 4 • · · · • · · · · · · · • · ·

219 ······ · · · · · ·

3α-methyl1-epimer + 33-methyl1-epimer

Hydrogenation of the mixed product prepared in step (e) (335 mg) as described in Example 166, step (g) and concentration of the resulting product from dioxane gave 469 mg of crude product as a gummy residue.

G)

3α-methyl-epimer + 3β-methyl-epimer

Treatment of the crude mixed product prepared in step (f) (460 mg) with N,N'-dicarbobenzyloxy-S-methylisothiourea and mercuric chloride according to the method of Example 166, step (h) gave, after chromatography on 75 g of silica gel using ethyl acetate:hexanes (6:4), 541 mg of a gummy residue containing the 3α-methyl epimer and the corresponding 3β-methyl epimer in the ratio (86:14).

220 • 0

h) Ιΐ

CH ₂ — O—C—N <Ì>~

CH2 _- OCN'

H ff

_{CH3.COCH3 ​}

3a-methyl-epimer and \-N

II ^H o

3β-methyl-epimer.

Treatment of the mixed product prepared in step (g) (500 mg) with sodium bis(trimethylsilyl)azide and phenyl isocyanate as described in Example 166, step (i), gave, after chromatography on 50 g of silica gel using ethyl acetate:hexanes (35:65), 525 mg of a gummy residue containing the 3α-methyl epimer and the corresponding 3β-methyl epimer in the ratio (86:14).

i) sP* ³ >p-OCH ₃ jj>H· 1.0 HC1~ II

_H2N ^NH

-N

O' _ _N

3a-methyl epimer JJ HO

3α-methyl-(epimer) (racemate) + 3β-πΐ6ί1ιγ 1-epimer

The mixed product prepared in step (h) (457 mg) was hydrogenated in 9 ml of dioxane with 0.73 ml of 1.0 N HCl in the presence of a catalyst, 10% palladium on carbon (160 mg) for 2 hours at 1 atmosphere. After filtration using dioxane, the filtrate was concentrated to a residue which was processed by lyophilization from aqueous

221 acetonitrile to 249 mg of white hygroscopic solid product containing 3α-methyl-epimer and corresponding 3β-methyl-epimer

in the ratio (88:12). IR (KBr) 1775 cm-1; (M+H) ⁺ 362. They were also prepared next with 1 lesson general formula III: NH SJ H ₂ NN R12 í i ^aRi H 0' Xj example Xi Rl R12 salt stereochemistry (M+H) ^{+ 169} í? -C-NH-(Î) ch ₃ 1.0 HCl racemate 415 -Lq 170 -CO2H ch ₃ — racemate 348 0 — C-NH-<^>

Example 171 (homochiral)

NH· 1 · OCF3COOH

A/x

O

II *COH

O

222

9 » • · « · • · * » 9 · ·

OH • ·« · »9

and)

O

II (H3O3—c—o-cHN. n>-COOH

Azetidine carboxylic acid is prepared from epichlorohydrin by the method described in A.G. Anderson Jr. and R. Lok, J. Org. Chem., Vol. 37, p. 3953 (1972). Said azetidine carboxylic acid is then converted to the desired product by the method described in WO97/23508 by Hansen T.L. et al.

b)

O

II (H3O3—c-o-οΧ

Br

To a stirred solution of the product prepared in step (a) (936 mg, 5 mmol) and tetrabromomethane (2.32 g, 7 mmol) in methylene chloride (910 ml) under argon at 0 °C was added dropwise a solution of triphenylphosphine (1.57 g, 6 mmol) in methylene chloride (6 ml). The reaction mixture was then stirred for 16 hours at room temperature. The reaction mixture was then concentrated in vacuo and the residue was triturated with ethyl ether. Filtration and evaporation of the filtrate gave 3.34 g of an oily residue, from which chromatography on silica gel using methylene chloride and then methylene chloride:ethyl acetate (19:1) as eluent gave 1.02 g of the desired product as an oily residue.

C)

O

II (H3O3-C-0-Cl

223 ·· 9* ·· · ·· • a· «*· *«· ···· · · * « «· • < · · · ······« « ··»··» 9 9

999* 99 99 9 99 9

A mixture of the product prepared in step (b) (1.00 g, 4 mmol) and sodium iodide (1.80 g, 12 mmol) in dry acetonitrile (10 mL) was stirred under argon at 65°C for 2.5 h and then concentrated in vacuo. The residue was taken up in ethyl acetate and water and the ethyl acetate layer was then washed with water (twice), dilute sodium thiosulfate, and water (twice), dried over sodium sulfate, and concentrated to give 1.18 g of the desired product as an oily residue.

d) iron (H ₃ C) ₃ — c— o- c—

C(CH ₃ ) ₃

To diisopropylamine (0.63 mL, 4.5 mmol) in tetrahydrofuran (3.5 mL) under argon at -20°C was added 1.6 mL of 2.5 M butyllithium in hexane (4 mmol). The resulting mixture was stirred for 15 min and then cooled to -70°C. A solution of (4S)-N-(tert-butyldimethylsilyl)azetidin-2-one-4-carboxylic acid (459 mg, 2.0 mmol) [Baldwin et al., Tetrahedron,

Vol.46, pp. 4733-4748, 1990] in tetrahydrofuran (2.5 ml) and the reaction mixture was warmed to -20 °C over 15 minutes. A solution of the product prepared in step (c) (1.19 g, 4 mmol) in tetrahydrofuran (3 ml) was then added and the reaction mixture was stirred at a temperature between -20 °C and 30 °C for 1.5 hours and then at -20 °C for 16 hours. The reaction mixture was then warmed to 0 °C and the reaction was quenched by the addition of 5% potassium hydrogen sulfate and then ethyl acetate. After extraction with ethyl acetate (three times), the ethyl acetate extracts were combined, washed with brine, dried over sodium sulfate and concentrated to an oily residue.

224 • · • » • · · • · · · 4

The residue was then dissolved in ethyl ether and washed with saturated sodium bicarbonate (twice). The combined sodium bicarbonate extracts were washed with ether and then layered with ethyl acetate. The pH was adjusted to 2.2 (10% potassium hydrogen sulfate) and after extraction with ethyl acetate (three times), the acidic ethyl acetate extract was washed with brine, dried over sodium sulfate and concentrated to give 624 mg of the desired product as a crude oil.

e) with

(H3O3— c— o-cCOOH

NH

To the crude product prepared in step (d) in tetrahydrofuran (3 ml) at 0-5 °C under nitrogen was added 2.7 ml of 1 M tetrabutylammonium fluoride in tetrahydrofuran. The reaction mixture was then stirred for 1.5 hours at room temperature, then the solvent was removed in vacuo, the residue was taken up in ethyl acetate, water and 10% potassium hydrogen sulfate (11 ml). After extraction with ethyl acetate (three times), the extracts were combined, washed with a small amount of water (twice), brine, dried over sodium sulfate and concentrated to give 470 mg of the crude desired product as an amorphous residue.

F)

O

II (H3O3— c— o-c-

» · · • · · · • · · · · ······

225 • · · · · c · « · · · ·

A solution of the product prepared in step (e) (466 mg), benzyl bromide (0.84 mL, 7.1 mmol) and sodium hydrogen sulfate (239 mg, 2.84 mmol) in dry dimethylformamide (4 mL) was stirred for 16 hours at room temperature under nitrogen. The reaction mixture was then diluted with ethyl acetate and water and the aqueous layer was extracted with ethyl acetate (twice). The ethyl acetate extracts were then combined, washed with dilute potassium hydrogen sulfate, water (twice) and brine, dried over sodium sulfate and concentrated to give 563 mg of an oil. Purification of this oil by chromatography on silica gel using methylene chloride followed by methylene chloride:ethyl acetate (1:1) as eluent gave 407 mg of the desired product as an amorphous residue.

To a stirred solution of the product prepared in step (f) (300 mg, 0.80 mmol) in methylene chloride (3 mL) was added trifluoroacetic acid (1 mL) at 0-5° C. After 5 minutes, the reaction mixture was stirred at room temperature for 1 hour and then concentrated in vacuo to give the desired product as a residue.

h)

226 ···· · · in · · · * »··

To a solution of the product prepared in step (g) (0.80 mmol) in dry dimethylformamide (3 mL) under argon was added Ν,N'-dicarbobenzyloxy-S-methylisothiourea (430 mg, 1.20 mmol), triethylamine (0.45 mL, 3.23 mmol), and mercuric chloride (326 mg, 1.20 mmol). The reaction mixture was then stirred at room temperature for 2.5 hours and then filtered through Celite ^R using ethyl acetate. The filtrate was washed with dilute aqueous potassium hydrogen sulfate (twice) and brine, dried over sodium sulfate, and concentrated to an oily residue (767 mg). Purification by chromatography on silica gel using methylene chloride:ethyl acetate (6:4) and methylene chloride:ethyl acetate (3:7) as eluent gave 342 mg of the desired product as an oily residue.

i) <2/ <2Ϊ

CH ₂ —OCN ch ₂ —O— C—

O

The product prepared in step (h) (136 mg, 0.23 mmol), which had been previously dried by azeotropic evaporation from tetrahydrofuran and toluene, in tetrahydrofuran (2 ml), was added to 0.30 ml of 1.0 M sodium bis(trimethylsilyl)amide (0.30 mmol) at -78 °C under argon. The resulting mixture was stirred for 10 min and then phenylisocyanate (31 μΐ, 0.28 mmol) was added. The reaction mixture was then warmed to 0 °C over 40 min and then poured into 2 ml of 10% potassium hydrogen sulfate and water. After extraction with ethyl acetate (three times), the ethyl acetate extracts were combined, washed with water (three times), dried over sodium sulfate and concentrated to give 193 mg of an oil. Purification by chromatography

227 • · * · ··· ······· · ····· · ·· · «· · on silica gel using a mixture of methylene chloride:ethyl acetate (98:2) and then methylene chloride:ethyl acetate (95:5) as eluent gave 121 mg of the desired product as an oily residue.

O

The product prepared in step (i) (115 mg, 0.163 mmol) was hydrogenated for 2 hours at 1 atmosphere in dioxane (4 ml) containing 1.0 N HCl (163 μΐ, 0.163 mmol) in the presence of a catalyst, 10% palladium on carbon (35 mg). After filtration using aqueous dioxane, the filtrate was concentrated to a residue. The residue was then lyophilized from aqueous dioxane to give 89 mg of crude product. Purification by HPLC [YMC S5 ODS 30 x 250 mm, 25 ml/min, using a gradient elution (10-40%) with solvent A (10% methanol + 90% water + 0.1% trifluoroacetic acid) and solvent B (90% methanol and 10% water + 0.1% trifluoroacetic acid)] gave 31 mg of the desired product as a hygroscopic solid after lyophilization. IR (KBr) 1780 cm-1; (M+H) ⁺ 346.

Example 172 (diastereomeric mixture)

1.0HCl'HN=

About aCOH

N- C- O— CH—· CH (CH ₃ ) 2 CH(CH ₃ ) ₂ • · ···· · v·· « · • · · · · ······* · • · · · · · · * ······ ·· · · ♦

228

and)

O

II (H3O3C— o-C-N

C-O-C2H5

To a stirred solution of ethyl nipecotate (6.2 mL, 40 mmol) and N,N-diisopropylethylamine (7.0 mL, 40 mmol) in methylene chloride (80 mL) at 0 °C under argon was slowly added

D-tert-butyl dicarbonate (10.9 g, 50 mmol). The cooling bath was then removed, dimethylaminopyridine (0.49 g, 4 mmol) was added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was then concentrated in vacuo and the residue was taken up in ethyl acetate. The ethyl acetate portion was then washed with dilute HCl (2 x), brine (2 x), dried over magnesium sulfate and concentrated to an oil which was passed through a silica gel column using hexane-ethyl acetate (8:2) to give 10.2 of the desired product as a colorless oil.

}—CH2-OH

N—' /

C=O

I am

AND

C-(CH ₃ ) ₃

To a stirred solution of the product prepared in step (a) (9.92 g, 38.5 mmol) in tetrahydrofuran (120 ml) at 0 °C under argon was added 1 M lithnoaluminum hydride (40.4 ml, 40.4 mmol) over 10 min. After 45 min, the ice-water cooled reaction mixture was quenched by the careful dropwise addition of 5 N sodium hydroxide (10 ml). The mixture was then stirred for 10 min and the semi-granular mixture was filtered. The filtrate was concentrated to an oil which was taken up in ether. The ether was then washed with brine, dried over sodium sulfate and concentrated to an oil which

229 • · solidifies in vacuum to 7.73 g of the desired product.

/

C=O and

O

AND

C-(CH ₃ ) ₃

A solution of the product prepared in step (b) (4.1 g, 20 mmol) in 30 mL of methylene chloride was added over 10 min to a stirred solution of triphenylphosphine (7.34 g, 28 mmol), imidazole (1.91 g, 28 mmol) and iodine (7.1 g, 28 mmol) in 70 mL of methylene chloride under argon at 0 °C. The cooling bath was then removed and the reaction mixture was stirred at room temperature for 1 h and then filtered. The filtrate was concentrated to give an oil which was stirred with ethyl acetate for 15 min. After filtration, the filtrate was washed with 5% sodium thiosulfate (3x) and then brine, dried over magnesium sulfate and concentrated to give 11.4 g of crude product, which was purified by chromatography on silica gel using methylene chloride to give 6.1 g of the desired product as a colorless oil.

C(CH ₃ ) ₃

Reaction of (4S)-N-(tert-butyldimethylsilyl)azetidin-2-one-4-carboxylic acid (1.15 g, 5 mmol) and the product prepared in step (c) (3.25 g, 10 mmol) according to the method of Example 171, step (d) gave 1.98 of the desired crude product as a foam.

230 products.

E)

_{to CO2H}

O

C(CH ₃ ) ₃

Treatment of the product prepared in step (d) (1.98 g) with tetrabutylammonium fluoride according to the method of Example 171, step (e) gave 1.64 g of the crude desired product as an oil.

F)

C(CH ₃ ) ₃

Treatment of the product prepared in step (e) (1.64 g) with benzyl bromide according to the method of Example 171, step (f), followed by chromatography on silica gel using methylene chloride and then methylene chloride/ethyl acetate (6:4) as eluent gave 1.23 g of the desired product as an oil.

G)

O

C(CH ₃ ) ₃ c ^x W ^r '' ^C “ ^O “f ^H ~ ^{CH(CH 3 ) 2}

About CH(CH ₃ ) ₂

231 ······ · · · · * ·

A solution of the product prepared in step (f) (402 mg, 1 mmol), triethylamine (0.28 mL, 2.0 mmol), i-diisopropylmethoxycarbonylpiperazine-4-carbonyl chloride (436 mg, 1.50 mmol) and dimethylaminopyridine (31 mg, 0.25 mmol) in methylene chloride (4.5 mL) was stirred under argon for 8 h at room temperature and then left overnight at 0 °C. The reaction mixture was then concentrated in vacuo and the residue was taken up in ethyl acetate, 10% potassium hydrogen sulfate (4 mL) and water. The ethyl acetate layer was washed again with dilute potassium hydrogen sulfate, water (2 x) and brine, dried over sodium sulfate and concentrated to a viscous oil (779 mg). Chromatography of this oil on silica gel using 10% and then 20% ethyl acetate in methylene chloride afforded 618 mg of the desired product as an oil.

To a stirred solution of the product prepared in step (g) (436 mg, 0.66 mmol) in methylene chloride (3 mL) was added trifluoroacetic acid (0.75 mL) at 0-5° C. After 5 minutes, the reaction mixture was stirred at room temperature for 2 hours and then concentrated in vacuo to a residue which was reconcentrated from methylene chloride (5 x) and then from chloroform (2 x) to give 648 mg of the title compound as an oil.

232 • · * · • · · · · · · · · · · ·

i) /—\ II (O/— CH ₂ —OCN

To a solution of the product prepared in step (h) (0.25 mL) in dimethylformamide (1.5 mL) under argon, Ν,Ν'-dicarbobenzyloxy-S-methylisothiourea (136 mg, 0.38 mmol), triethylamine (0.21 mL, 1.5 mmol) and mercuric chloride (103 mg, 0.38 mmol) were added sequentially. The reaction mixture was stirred at room temperature for 3 hours, diluted with ethyl acetate and filtered through Celite. The filtrate was then washed with dilute aqueous potassium hydrogen sulfate (2 ×), brine (2 ×), dried over sodium sulfate and concentrated to an oil (280 mg) which was purified by chromatography on silica gel using 15% and then 20% ethyl acetate in methylene chloride to give 146 mg of the desired product as an oily residue.

(diastereomeric mixture)

O-CH—CH(CH ₃ ) ₂ CH(CH ₃ ) ₂

The product prepared in step (i) (141 mg, 0.163 mmol) was hydrogenated for 1 hour at 1 atmosphere in dioxane (5 mL) containing 1.0 N HCl (0.163 mmol) in the presence of a catalyst,

233

10% palladium on carbon (42 mg). After filtration using dioxane, the filtrate was concentrated to remove dioxane, filtered and lyophilized to give 47 mg of the desired product as a white solid. IR (KBr) 1787 cm ^-1 , HPLC analysis indicated that the product contained a mixture of diastereomers (52:48).

Other compounds of general formula I were also prepared:

example -Ai- Xi

173

N.

CH ₂

Ri salt stereochemistry (M+H) ⁺

-CO2H --- homochiral 481

CH-CH(CH ₃ ) ₂

CH(CH ₃ ) ₂

ch ₃

1.0 HCl homochiral 452

174

234 example -Ai~ Xi

Ri salt stereochemistry (M+H) ⁺

175

1.0 HCl homochiral 374

177

CH ₂

-CO2H 1.0 HCl diastereomeric mixture 495

O

II

-cN— C- O- CH—- CH (CH ₃ ) ₂ CH(CH ₃ ) ₂

Example 178 (homochiral)

NH to

H ₂ N N> H roH fí •N

O'>c— N NCO— CH-CH(CH ₃ ) ₂ \ / ¹ '— ^f CH(CH ₃ ) ₂

235 • · · · · • · · · · · · • ·······

a) (H3O3-C-O-C·

To a solution of di(tert-butoxycarbonyl)amine (2.17 g, 10 mmol) in dimethylformamide (40 ml) at 0 °C was added a solution of 6.8 g of potassium tert-butyl amylate in toluene. The addition was carried out very slowly over 30 minutes. The reaction mixture was then stirred for one hour at 0 °C, then 1-chloro-4-iodobutane (2.18 g, 10 mmol) was added dropwise and stirring was continued for another two hours. For further work-up, water (20 ml) and hexane (20 ml) were added. The aqueous layer was extracted with additional hexane (3 x 20 ml). The combined organics were washed with 1.0 N ice-cold sodium hydroxide, saturated sodium dihydrogen phosphate solution, and finally brine. After drying over sodium sulfate and evaporation, 2.76 g of the desired product was obtained as a pale yellow oil.

^b)

II (H ₃ C) 3—c—O—c·

To a solution of the product prepared in step (a) (4.5 g, 15 mmol) in acetone (60 ml) was added sodium iodide (7.3 g, 49 mmol) and sodium bicarbonate (12 mmol). The reaction mixture was then heated under reflux at 60 °C for 12 hours. Additional sodium iodide (2 g) and acetone (30 ml) were then added and heating at reflux was continued for a further 8 hours. The reaction mixture was then evaporated, the residue (oil and solid) was extracted with hexane (5 x 30 ml), the combined extracts were washed with 2.0 N sodium sulfite and brine, and

236 « · « · · · · · · · · · • · · · · ·········· Drying with sodium sulfate and concentration afforded 5.02 g of the desired product as a yellow oil.

······ ·· · ·· ·

c) (H3C) 3—C—O—

(4S)-N-(tert-butyldimethylsilyl)azetidin-2-one-4-carboxylic acid (2.30 g, 10 mmol) [Baldwin et al., Tetrahedron, Vol.46, pp.4733-4748, 1990] was dissolved in tetrahydrofuran (10 ml) and added dropwise over 20 minutes at -50°C to a solution of 1-lithiumdiisopropylamide (21 mmol) in tetrahydrofuran (60 ml). After the addition was complete, the reaction mixture was allowed to warm to -20°C, at which point a solution of the product prepared in step (b) (4.80 g, 12 mmol) in tetrahydrofuran (10 ml) was added dropwise. Stirring was continued for two hours at -20°C after the addition was complete. The reaction mixture was then warmed to 0°C, water (100 ml) was added and the pH was adjusted to 12.5 by adding 1.0 N sodium sulfate.

After stirring for 1 hour at 0°C, the reaction solution was extracted with hexane (50 mL). The pH of the aqueous layer was then adjusted to pH 3 with 6.0 N HCl, and the aqueous layer was extracted with ethyl acetate (150 mL). The organic phase was then washed with brine and dried to give 4.13 g of the desired product as a colorless oil.

d) < O )-CH ₂ — OC—N ©— ch ₂ — o- c-ř/^

237 • · · · ·

To the product prepared in step (c) (2.5 g, 5 mmol) dissolved in methylene chloride (50 ml) at 0 °C was slowly added trifluoroacetic acid (25 ml). The reaction mixture was then stirred for 30 minutes, evaporated and toluene (20 ml) was added to the oily residue. The toluene was then evaporated again to remove excess trifluoroacetic acid to give 1.6 g of the compound of formula

which is used in the next step without purification.

The above trifluoroacetic acid salt (1.6 g) was dissolved in methanol (30 ml), cooled to -5 °C, then the pH was adjusted to 8.5 by addition of triethylamine (0.7 ml) and then Ν,N'-dicarbobenzyloxy-S-S-methylisothymourea (2 g, 5.5 mmol) was added. The reaction mixture was then stirred for 12 hours at room temperature. The methanol was then evaporated in vacuo and the oily residue was taken up in ethyl acetate (20 ml) and water (10 ml). The mixture was then cooled to 0 °C, the pH was adjusted to 3.0 with 2.0 N sodium hydrogen sulfate solution. The layers were separated and the aqueous layer was extracted with ethyl acetate (2 x 20 ml). The combined organics were washed with brine and extracted with ice-cold saturated sodium bicarbonate solution (3 x 20 ml). After cooling to 0°C, the aqueous phase was acidified with concentrated HCl to pH 3.2 and re-extracted with ethyl acetate (3 x 20 mL). The combined organic layers were washed with brine, dried over sodium sulfate and concentrated to give 1.95 g of the desired product as a colorless oil.

238

E)

CH ₂ —OC—N « A

CH ₂ — oCN

The product prepared in step (d) (497 mg, 1.0 mmol) was dissolved in tetrahydrofuran (5 ml) and butanol (155 μΐ, 1.5 mmol), dicyclohexylcarbodiimide (210 mg, 1.0 mmol), 4-dimethylaminopyridine (10 mg) and hydroxybenzotriazole (20 mg) were added with stirring. After stirring for 6 hours at room temperature, methylene chloride (5 ml) was added. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give 570 mg of crude product as a colorless oil. Purification by flash chromatography on silica gel using ethyl acetate/hexane gave 495 mg of the desired product. IR (KBr) 1745 cm-1.

f) <2><27

II ch ₂ —o—what

II

CH ₂ — oCN

The product prepared in step (e) (295 mg, 0.5 mmol) was dissolved in methylene chloride (7 mL). Diisopropylamine (194 mg, 1.5 mmol), 1-diisopropylmethoxycarbonylpiperazine-4-carbonyl chloride (218 mg, 0.75 mmol) and dimethylaminopyridine (10 mg) were added and the reaction mixture was stirred overnight at room temperature.

239 • ·

room temperature. Additional 1-diisopropylmethoxycarbonylpiperazine-4-carbonyl chloride (20 mg) was then added and stirring was continued for another 4 hours. The reaction mixture was then quenched with 10 mL of ice water (pH adjusted to 4 with potassium sulfate solution) and ethyl acetate. The aqueous layer was then extracted with ethyl acetate (2 × 10 mL) and the combined organics were washed with brine, dried over magnesium sulfate and concentrated in vacuo. The resulting colorless oil was purified by flash chromatography on silica gel using ethyl acetate:hexanes (4:6) as eluent to give 392 mg of the desired product. IR (KBr) 1790 cm ^-1 .

G)

NH

_H2N ,

C-OH (homochiral) c—N NCO—CH-CH(CH ₃ ) ₂ v_/

CH(CH ₃ ) ₂

A mixture of the product prepared in step (f) (96 mg, 0.11 mmol), 1 N HCl (110 μΐ, 0.11 mmol) and the catalyst, 10% palladium on carbon (37 mg) in dioxane (2.5 ml) was stirred for 1 hour under hydrogen atmosphere (under a hydrogen balloon). The reaction mixture was then filtered through a pad of Celite ^R , passed through a column of polyvinylpyrrolidone resin, and lyophilized to give 44 mg of the desired compound as a white flaky powder. IR (KBr) 1780 cm ^-1 , 1669 cm ^-1 ; (M+H) ⁺ 483.3;

(M-H)- 481.

240

·· ·« » · · • · · ·

4· * « · · • · · · • · · · · ·*·· ··

In addition to the compounds prepared by Han in U.S. Patent No. 5,037,819, other compounds of general formula VI have been prepared by Han:

NH η ₂ ν ^Λ ν^<... ' ^Ru

H for

N

X3

example X3 Rl 4 salt with stereochemistry (M+H) 179 _-CO2H 1.0 CF3CO2H homochiral 319 180 0 -every ₂ hours 1.0 HCl homochiral 334 -“Γ® 181 0 -CO2CH3 1.0 HCl homochiral 348 182 0 -CO2H 1.0 CF3CO2H racemate 33 --r®

241 • · ·* 9 » # ·« * · · ·«· *, · • · · · « »·· · · • * ··« ··>«··* » •44494 4 4 9 9 9

example X3 Rl 4 salt stereochemistry (M+H) 183 _Λ 1.0 HCl -'-•O racemate 401 184 0 II -C—CH3 1.0 HCl ~(CH ₂ , ₂ -^) homochiral 317 185 --šÁ2> _ 1.0 HCl -(CH ₂ ) ₂ -ïY> racemate 394

In addition, other compounds of general formula VI were prepared:

NH

HER

H ₂ NN' H ^P 14

X3

242 ·· ft· ·· » ·· ··· *«· 1 · ·«·· · » * · · · • * « · » · ···· · · · • » · · f · ·· ···· ·· ·· · »· ·

example X3 R 1 4 salt stereochemistry (M+H) 186 — (CH ₂ ) ₂ — 1.0 HCl racemate 394 187 0 li -c—ch ₃ — (CH ₂ ) ₂ — 1.0 HCl racemate 317

In addition, other compounds of general formula VI were prepared:

_H2N

NH

H ^14

N

243 ··· ··« · · · » ···· to · · · · · ♦ • · · · to ····»·· « · ··· ··· ··· ······ · · · · · t · J

example _x3 Rl 4 salt with stereochemistry (M+H) 188 0 -C-NH-^O^ -CO2CH3 1.0 CF3CO2H homochiral 348 189 — C-NH-^O^ -CO2H homochiral 332 In addition, formulas VI were prepared: NH JI H ₂ NN ^/X ^ ⁵ o' other compounds of general formula W" -N X ₃ example X3 Rl4 salt with stereochemistry (M+H) 190 0 — C-NH-{O^ -CO2CH3 1.0 HCl racemate 348 191 — C—NH-^O^ -CO2H 1.0 CF3CO2H racemate 334

244 • · · · ··· ·· · ··· · · · · · *» • · · · · ··· · J · • · ··· ······· β * • · · ··· · · · ···· ·· ·· · ··»

Example 192

The intermediate of Example 1(b) can also be prepared as follows:

a) (H ₃ C) ₃ -C—O—C--N-(CH ₂ ) ₃ —Cl

To a solution of di-tert.butyliminodicarboxylate (70.1 g,

322.6 mmol) in dimethylformamide (500 mL) was added potassium tert-amylate (25 wt% in toluene, 164.0 g, 322.6 mmol) over 15 min under nitrogen. The resulting creamy white solution was stirred at 0 °C for 40 min. 1-Chloro-3-iodopropane (60.0 g, 31 mL, 293.3 mmol) was then added and the mixture was stirred at 0 °C for 3 h. Hexane (500 mL) and water (300 mL) were then added to the mixture. The aqueous layer was separated and extracted with hexane (300 mL). The combined hexane extracts were washed with 1N sodium hydroxide (3 x 300 mL), saturated sodium hydrogen phosphate (300 mL), half saturated brine (300 mL), and brine (500 mL) and dried over sodium sulfate. Removal of sodium sulfate by filtration and subsequent concentration afforded 84.2 g of the desired product as a pale yellow oil, which was further dried overnight in vacuo.

245 • 0

000 000 0 0 0 0000 0 000 0 0 ·

0 040 0000000 0 0

In 0 ·00 0 · ·

000000 00 0 00 000

b) (H3O3-COC·— n-(ch ₂ ) ₃

The product prepared in step (a) (55.0 g, 187.2 mmol) was dissolved in acetone (550 ml). Sodium iodide (84.2 g,

561.6 mmol) and sodium bicarbonate (7.9 g, 93.6 mmol). The resulting mixture was stirred at 57 °C (oil bath) under nitrogen for 6 hours, and then additional sodium iodide (14 g, 93.4 mmol) was added. The reaction mixture was stirred for 12 hours, and then acetone was evaporated. Hexane (400 mL) and water (300 mL) were added to the resulting solid. The hexane layer was separated, washed with 5% sodium thiosulfate (300 mL), half-saturated brine (300 mL), and brine (300 mL), dried over sodium sulfate, and filtered through a 20 g pad of silica gel. The silica gel pad was then washed with hexane (400 mL). Concentration of the filtrate gave 68.2 g of the desired product as a pale yellow oil, which was then dried overnight in vacuo.

c) (H ₃ C) ₃

COOH •NH

To a solution of diisopropylamine (11.7 g, 16.2 mL, 115.3 mmol) in tetrahydrofuran (150 mL) under nitrogen at 0 °C was added butyllithium (2.5 M in hexane, 46 mL, 115.0 mmol).

The resulting solution was stirred for 30 minutes at 0°C and then cooled to -30°C. A solution of (4S)-N-(tert-butyldimethylsilyl)azetidin-2-one-4-carboxylic acid (12.0 g, 52.4 mmol) [Baldwin et al., Tetrahedron, Vol.46, pp.4733-4748, 1990] in

246 • a · · · ······· • · · · · f · • * · · «· ·· · tetrahydrofuran (60 ml) and the mixture was stirred at -20 °C for 30 minutes. Then the iodine derivative prepared in step (b) (24.0 g, 62.3 mmol) in tetrahydrofuran (30 ml) was added dropwise over 20 minutes and the resulting mixture was stirred at -20 °C for 2 hours. Then the reaction mixture was allowed to warm to 0 °C and water (300 ml) was added. Then the pH of the mixture was adjusted to 12.5 with 10% sodium hydrogen sulfate, stirred at 0 °C for 30 minutes and then washed with hexane (2 x 100 ml). The aqueous layer was cooled in an ice-water bath and acidified to pH 3.0 by dropwise addition of 6 N HCl. This solution was saturated with sodium chloride and extracted with ethyl acetate (3 x 150 mL). The combined ethyl acetate extracts were washed with brine (200 mL), dried over sodium sulfate, filtered, and concentrated to a yellow oil. The resulting oil was dissolved in acetonitrile (20 mL), and the acetonitrile was evaporated to give 16.4 g of the desired product as a yellow foam.

<ΖΟ ₂ Η hi,.

/-NH

O

The product prepared in step (c) (4.25 g, 11.4 mmol) was added to a 1:2 mixture of trifluoroacetic acid/methylene chloride (42 mL). The resulting mixture was stirred for 30 minutes under nitrogen at room temperature. Toluene (80 mL) was then added and the mixture was concentrated to a small volume (ca. 15 mL). Additional toluene (80 mL) was then added and the mixture was concentrated to dryness to give a yellow oil.

To the above oil was added a mixture of triethylamine (3.97 mL, 28.5 mmol) and methanol (42 mL) at 0 °C. Additional triethylamine (0.85 mL, 5.7 mmol) was then added and

247

N,N'-bis(benzyloxycarbonyl)-1-guanylpyrazole (4.31 g, 11.4 mmol) [Wu et al., Synthetic Communications, 23(21) pp. 3055-3060 (1993)]. The resulting mixture was then stirred at room temperature for 11 hours, and then concentrated in vacuo at 25°C to give a yellow oil.

To the oil was added ethyl acetate (30 mL) and water (10 mL), the mixture was acidified at 0°C to pH 3.2 by addition of 2 M potassium hydrogen sulfate saturated with sodium chloride. The acidic mixture was then poured into a separatory funnel. The layers were separated, the aqueous layer was washed with ethyl acetate (2 x 25 mL) checking that the pH of the aqueous solution was 2.9 to 3.2 in solution. The combined ethyl acetate solutions were washed with saturated sodium chloride solution (25 mL) and the product was extracted with saturated sodium bicarbonate (3 x 25 mL). The combined bicarbonate solutions were washed with ethyl acetate (2 x 25 mL), acidified to pH 3.2 at 0 °C with concentrated HCl, treated with saturated sodium chloride (solid) and finally extracted with ethyl acetate (3 x 25 mL). The combined ethyl acetate solution was dried over magnesium sulfate, filtered and concentrated in vacuo to give 4.61 (9.55 mmol) of the desired product as a pale yellow foam.

Example 193

The products of Examples 21 and 32 are also prepared as follows:

and)

JN—Si(CH ₃ ) ₂ o I

C(CH ₃ ) ₃

248

A dry three-necked flask equipped with a mechanical stirrer was charged with diisopropylamine (14.05 mL, 0.10 mmol) and maintained under argon. Tetrahydrofuran (anhydrous, 33 mL) was then charged to the flask and, while stirring and cooling the mixture to -20°C, a solution of butyllithium (38.3 mL of a 2.5 M solution in hexanes, 0.096 mol) was added dropwise and the solution was then stirred for 10 minutes at -20°. A solution of (4S)-N-(tert-butyldimethylsilyl)azetidin-2-one-4-carboxylic acid (10.0 g, 43.6 mmol) [Baldwin et al., Tetrahedron, Vol.46, pp.4733-4748, 1990] was then slowly added while maintaining the temperature at -20°C and the reaction mixture was stirred for 30 minutes. A solution of 1-chloro-3-iodopropane (5.6 mL, 52 mmol, 1.2 eq.) in tetrahydrofuran (30 mL) was then added over about 10 minutes. The reaction mixture was then stirred for about 2 hours at -20 °C and then 2.6 M potassium hydrogen sulfate (75 mL), water (100 mL) and ethyl acetate (100 mL) were added and the mixture was transferred to a separatory funnel. The aqueous layer (pH 2-3) was drained and back-extracted with ethyl acetate (2 x 50 mL). The organic solutions were combined and washed sequentially with water (2 x 50 mL), 10% sodium thiosulfate (1 x 50 mL) and then saturated sodium chloride to neutral pH. The organic solution was then dried over sodium sulfate (15 g), filtered and concentrated to an oil. The oil was seeded with a few crystals of the desired product and dried under high vacuum overnight. The resulting crystalline solid was slurried with hexane (100 mL), filtered, washed with hexane (100 mL), and dried under high vacuum to give 11.5 g of the desired product.

b) _aCO ₂ CH ₂ —

JN—Si(CH ₃ ) ₂

C(CH ₃ ) ₃ • · · · ·« « ·

249

The product prepared in step (a) (7.02 g, 22.9 mmol) was dissolved in methylene chloride (anhydrous, 40 mL) under argon. The resulting solution was cooled to 0 °C with stirring and triethylamine (3.5 mL, 25.2 mmol) was added slowly, maintaining the temperature at about 0 °C. Benzyl chloroformate (3.6 mL, 25.2 mmol) was then added along with an additional 10 mL of methylene chloride to facilitate stirring. 4-dimethylaminopyridine (2.8 g, 22.9 mmol) was then added in one portion as a solid, with significant evolution of gas. The solution was then stirred at 0 °C for 30 min. Additional benzyl chloroformate (0.3 mL, 2.5 mmol) was then added and the reaction mixture was stirred for a further 20 min. The reaction mixture was then quenched with 1 M potassium hydrogen sulfate (30 mL). The mixture was then transferred to a separatory funnel and the layers were separated. The organic layer was washed with 2 N potassium hydrogen sulfate (20 mL), the aqueous washings were combined and re-extracted with methylene chloride (25 mL). The organic solutions were combined, washed successively with water (25 mL), saturated sodium bicarbonate (25 mL), and saturated sodium chloride (2 x 25 mL), and dried over sodium sulfate (15 g). The methylene chloride solution was filtered through silica (15 g) and the silica was washed with 200 mL of hexane/ethyl acetate 3:1 (v/v). The filtrate was concentrated to an oil, evaporated under reduced pressure with toluene (2 x 25 ml) and dried in vacuo overnight to give 8.12 g of the desired product.

C)

N—Si(CH ₃ ) ₂ C(CH ₃ ) ₃

A solution of the product prepared in step (b) (10.55 g, 26.6 mmol) in 4-methyl-2-pentanone (40 ml) was stirred under an atmosphere of

250 argon. Sodium iodide (20 g, 133 mmol) was then added and the mixture was heated in the dark at 110°C for about 7 hours. The mixture was then cooled to room temperature, diluted with hexane (100 ml) and filtered through a pad of Celite ^R . The Celite ^R pad was then washed with hexane (2 x 50 ml). The filtrates were combined, washed with sodium thiosulfate (50 ml) and then water (50 ml). The aqueous washings were re-extracted with ethyl acetate (100 ml). The organic solutions were combined, concentrated to an oil, then dissolved in hexane (50 ml) and filtered through a pad of silica. The silica was then washed with hexane (200 ml) and then with a 4:1 (v/v) hexane/ethyl acetate mixture (500 ml).

The hexane/ethyl acetate solution is then concentrated to 11.84 g of the desired product, which is in the form of an oil.

(5 g, 20.5 mmol) and tetrahydrofuran (27 mL, anhydrous) were charged to a dry flask under nitrogen. The resulting solution was stirred and cooled to 0 °C. Sodium hydride (2.62 g, 65.5 mmol, 3.2 equivalents of a 60% dispersion in mineral oil) was then slowly added to the flask (exothermic). The suspension was stirred at 0 °C. Then, sodium hydride (2.62 g, 65.5 mmol, 3.2 equivalents of a 60% dispersion in mineral oil) was added portionwise.

N-(benzyloxycarbonyloxy)succinimide (8.2 g, 32.8 mmol, 1.6 equiv.) while maintaining the temperature at about 0°C. Cooling was then removed and the reaction mixture was allowed to warm to room temperature.

/

251 « · * · ♦ *- · · · • · · ····· • » · f

After 1 hour, another portion of N-(benzyloxycarbonyloxy)succinimide (1 g, 0.2 eq.) was added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was then cooled to about 0 °C and slowly quenched with 13% aqueous ammonium chloride. The layers were separated and the aqueous layer was re-extracted with ethyl acetate (3 x 20 mL). The combined organic solution was washed successively with 13% aqueous ammonium chloride (5 mL), water (15 mL), and saturated sodium chloride (2 x 15 mL). The organic solution was then dried over sodium sulfate, filtered, and concentrated. The resulting crude oil was charged to a flask with 2 M ammonia in methanol (51 mL, 101 mmol, 5 eq.) and stirred overnight at room temperature. The reaction mixture was then concentrated under reduced pressure and then coevaporated with hexanes (2 x 25 mL). The residue obtained was dissolved in methylene chloride (50 mL) and washed with water (50 mL). The aqueous layer was then re-extracted with methylene chloride (2 x 50 mL). The organic extracts were combined, washed with water (25 mL) and saturated sodium chloride (25 mL), dried over sodium sulfate, filtered, and concentrated to a solid. Crystallization of this residue from ethyl acetate afforded 4.65 g of the desired product as white crystals.

E)

O

The product prepared in step (d) (6.01 g, 18.37 mmol) was dissolved in 1-methyl-2-pyrrolidinone (anhydrous, 10 ml) and heated under argon to 35-40 °C. Potassium carbonate (finely ground and dried, 12.7 g, 92 mmol) was then added and the mixture was stirred for 25 minutes. The product prepared in step (c) was then added.

252 • · fl (9.5 g, 18.37 mmol) in 1-methyl-2-pyrrolidone (10 mL) and the reaction mixture was stirred for 8 hours at 35-40 °C. The reaction mixture was then cooled to room temperature, diluted with ethyl acetate (100 mL) and filtered into a separatory funnel. The mixture was then washed with 1 M potassium hydrogen sulfate (84 mL) and the layers were separated. The aqueous layer was then extracted with ethyl acetate (50 mL). The organic solutions were combined, washed successively with water (50 mL), 10% sodium thiosulfate (50 mL) and saturated sodium chloride (50 mL), dried over sodium sulfate, filtered and concentrated to an oil. The oil was dissolved in 50 mL of a 1:1 (v/v) hexane/ethyl acetate mixture and filtered through silica. The filtrate was concentrated to give 12.88 g of the desired product as a crude oil which was used without further purification.

The crude oil prepared in step (e) was dissolved in acetonitrile (50 mL) and water (5 mL). Ammonium fluoride (3.4 g, 92 mmol) and glacial acetic acid (5.25 mL, 92 mmol) were then added and the mixture was stirred for 30 min. The reaction mixture was then diluted with ethyl acetate (150 mL), transferred to a separatory funnel and washed with saturated sodium hydrogen sulfate (30 mL). The layers were then separated and the aqueous layer was re-extracted with ethyl acetate. The organic solutions were combined, washed with saturated sodium chloride, dried over sodium sulfate, filtered and concentrated to an oil. The oil was then dissolved in ethyl acetate (30 mL) and heated to 60 °C. Hexane was then added and the mixture was allowed to cool slowly while stirring and seeded. When crystallization begins, hexane (75 ml) is added in portions. The solid obtained is

253 « 9 · · · ·

9 · · ♦ 8 « *

filtered off, washed with hexane and dried to give 8.91 g of the desired product.

G)

About About

Cl— C— C—NH— C (CH ₃ ) ₃

In a dry 2-liter flask equipped with a mechanical stirrer and an argon inlet, 1-benzylpiperazine (40 mL, 230 mmol) and toluene (250 mL) were placed. Then, with vigorous stirring, tert-butyl isocyanate (27 mL, 236 mmol) was added dropwise rapidly over 15 minutes. A thick slurry containing the product was formed. The slurry was stirred for one hour to bring its temperature to 25 °C. Heptane (570 mL) was then added to the slurry over 30 minutes. The flask was then capped and placed in a cooling chamber (5 °C) for 4 hours. The product was then filtered off, washed with heptane (1 x 200 mL) to give 61.0 g of piperazine of the formula

O

II

C— NH-C(CH ₃ ) in the form of a white solid product.

Methanol (200 mL) was charged into a 500 mL flask equipped with a magnetic stirrer and a gas introduction tube. The flask was then cooled to 1°C and then acetyl chloride (5.7 mL, 79.9 mmol) was added over 10 min with stirring. The solution was allowed to warm to room temperature and then piperazine (20.0 g, 72.6 mmol) was added. Palladium hydroxide on carbon (8.0 g, water content 50% or less) was added and argon was bubbled into the mixture for 10 min.

Hydrogen is then introduced into the reaction mixture. After 3.5 hours, HPLC indicates that all of the starting material has reacted. The mixture is then

254 is filtered through Celite ^R and the Celite ^R is washed with methanol (60 ml). The filtrate is concentrated until a solid product begins to crystallize (175 ml of methanol is separated). Isopropyl alcohol (200 ml) is then slowly added with manual stirring. The resulting mixture is concentrated to a solid/liquid mixture (weight 153 g). This mixture is left undisturbed for 2 hours. The product is then filtered off, washed with isopropyl alcohol (1 x 30 ml) and air-dried to give 14.0 g of the hydrochloride salt of the formula

HC1»HN N—

O

II

C— NH-C(CH ₃ ) in the form of a yellow solid product.

In a 100 mL three-necked flask equipped with a magnetic stir bar and a gas introduction tube, the above hydrochloride salt (5.0 g, 22.6 mmol) and anhydrous methylene chloride (50 mL) were charged. 1,8-diazabicyclo[5.4.0]undec-7-ene (6.7 mL, 45.1 mmol) and pyridine (1.8 mL, 22.6 mmol) were then added to the mixture. The mixture became homogeneous. Dry carbon dioxide gas was then introduced into the mixture for 1 hour at room temperature.

In a dry 500 ml flask equipped with a magnetic stir bar, thionyl chloride (4.9 ml, 67.7 mmol) and anhydrous methylene chloride (25 ml) were charged. The solution was then cooled to -10 °C and dimethylformamide (0.17 ml, 2.26 mmol) was added. The above mixture, saturated with carbon dioxide, was then added over 35 min via cannula and under carbon dioxide pressure. The flask was then washed with methylene chloride (5 ml) and the washings were added to the reaction mixture. The reaction mixture was then stirred for 30 min at -10 °C. Then • · · • * · · · · · · I

The reaction mixture was poured into 0.5 M HCl (75 mL) and shaken vigorously. The organic layer was separated, dried over magnesium sulfate, filtered, and concentrated in vacuo to give 4.95 g of the desired product as a pale yellow solid.

O

The product prepared in step (f) (11.84 g, 20.7 mmol) was dissolved in anhydrous methylene chloride (100 mL) with stirring under argon. The carbamoyl chloride product prepared in step (g) (7.47 g, 26.9 mmol, 1.3 eq.), triethylamine (4.6 mL, 33.1 mmol, 1.6 eq.), and 4-dimethylaminopyridine (0.76 g, 6.2 mmol, 0.3 eq.) were then added, and the reaction mixture was stirred overnight at room temperature. The reaction mixture was then poured into 0.5 N HCl (110 mL), the layers were separated, and the organic layer was washed with another portion of 0.5 N HCl. The acidic aqueous layers were back-extracted with methylene chloride (50 mL) and combined with the main organic portion. The combined organic layers were washed with saturated sodium bicarbonate (100 mL) and saturated sodium chloride, dried over sodium sulfate, filtered, and concentrated to give 17.0 g of the desired product as a crude white solid.

256 • 0 · 0 000 · · 000 000 000

0000 0 000 · · « 000 0000000 0

00000· 00 · ·· ·

and)

_H2N

O

NC—N—C(CH ₃ ) ₃ H

The crude product prepared in step (h) (17.0 g) was dissolved in absolute ethanol (350 ml) with stirring. Argon was then introduced into the mixture and the catalyst, 10% palladium on carbon (1.7 g, 50 wt. % water) was added in one portion and argon was again introduced into the mixture. Hydrogen was then introduced into the mixture for 2 minutes and then the mixture was placed under a pressurized hydrogen atmosphere (balloon). Two more portions of palladium on carbon catalyst (1.7 g each) were then added to the reaction mixture, repeating the washing procedure. After about 4 hours, the reaction was considered complete (HPLC analysis). Argon was then introduced into the reaction mixture for 5 minutes and then the mixture was filtered through a pad of Celite ^R.

The Celite ^R was then washed with ethanol (2 x 125 ml). The combined ethanol filtrates were then concentrated to about 50 g and the concentrate was stirred for 4 days. Crystals formed at the bottom of the flask. These crystals were filtered off, washed with absolute ethanol (25 ml) and dried to give 7.74 g of the desired product as a white crystalline mass. This product was further purified by heating in 95% ethanol at about 40 °C for 30 minutes, followed by cooling, filtration and drying.

Example 194

The products of Examples 21, 32 and 193 were also prepared in the form of zwitterionic or inner salts as follows:

• ·

257

a) (H3O3-C-O-

_to CO ₂ H«NH ₂ C(CH ₃ )3

Into a dry, 12-liter three-necked flask (flask A) are charged 769.4 g of (4S)-N-(tert-butyldimethylsilyl)azetidinonecarboxylic acid and then 6 liters of dry tetrahydrofuran. Into another 5-liter three-necked flask (flask B) are charged 1537.8 g of the iodine derivative prepared according to Example 192, step (b), and then 2 liters of dry tetrahydrofuran. Into a 22-liter flask (flask C) are charged 4 liters of dry tetrahydrofuran and then 3.69 liters of lithium diisopropylamide under a nitrogen atmosphere.

The solution of 1 lithium diisopropylamide is cooled to -30 to -40 °C. Then the contents of flask A are introduced into flask C, while maintaining the temperature below -20 °C. The resulting mixture is stirred at -20 to -25 °C for about 30 to 60 minutes and then cooled to -35 °C to -40 °C. The contents of flask B are then added in portions over 25 to 45 minutes to flask C, while maintaining the internal temperature in flask C at about -20 °C. The resulting mixture is stirred for 2 to 3 hours at a temperature in the range of -20 °C to -23 °C. Then the reaction mixture is poured with cold water so that the internal temperature of the batch ranges from -20 °C to +5 °C. The mixture was then stirred for an additional 15 to 30 minutes and then the pH was adjusted to 8.0 by the addition of cold 6N HCl (1.69 L) to remove the silyl protecting group. The reaction mixture was then transferred to a phase separator and the upper organic layer was removed. The aqueous layer was washed twice with 4 L portions of hexane. The aqueous phase was then cooled to about 0 °C and treated with 6N HCl (about 400 mL) to a pH of 3.0. The batch temperature was maintained below 5 °C during this treatment. The cloudy aqueous phase was then extracted three times with 4 L portions of ethyl acetate. The combined organic extracts were then washed with brine (3 x 3 L) and concentrated to an oil. The oil

258 • · · · · · · · * ·

To a suspension of the product prepared in step (a) (10.0 g, 22.44 mmol) in methylene chloride (40 mL) at -5°C to +5°C under nitrogen was added trifluoroacetic acid (31.1 mL, 403.9 mmol, 18 equivalents). The resulting pale yellow clear solution was stirred at 0°C until less than 1% of the mono-tert-butoxycarbonyl intermediate was detected by HPLC (about 4 to 7 hours). The methylene chloride and trifluoroacetic acid were then removed in vacuo at room temperature. To the residue was added toluene (30 mL) and then removed in vacuo. The residue was then treated with isopropyl alcohol (10 mL) and then toluene (20 mL) and the resulting solution was concentrated in vacuo to an oil. This step was repeated once more.

Isopropyl alcohol (50 ml) was then added to the above oil (about 23 mg) and the resulting solution was cooled to 0°C. The pH was adjusted to 8.5 to 9.0 at -5°C to 5°C using triethylamine added dropwise (use

259 about 18 ml of triethylamine). Then add in one portion

N,B'-bis(benzyloxycarbonyl)-1-guanylpyrazole (8.07 g, 21.32 mmol, 0.95 equivalents). The resulting mixture was stirred under nitrogen at room temperature for about 30 minutes until the product/pyrazole ratio was greater than 25:1 as determined by HPLC. The solvent was then removed in vacuo at room temperature to give about 42 g of a yellow oil. This oil was diluted with ethyl acetate (70 mL) and water (70 mL) and then cooled to 0°C. The pH of the solution was adjusted to 3.0 with 3.0 M potassium hydrogen sulfate and the solution was then treated with sodium chloride until saturated. The organic layer was then separated and the aqueous layer was extracted with ethyl acetate (2 x 60 mL). The combined ethyl acetate fractions were washed with saturated sodium chloride solution (2 x 60 mL), dried over sodium sulfate and filtered. The solvent was then evaporated to give a yellow oil (14.4 g) which was then redissolved in ethyl acetate (40 ml). The resulting clear yellow solution was then warmed to 36-40 °C and treated dropwise with tert-butylamine (3.3 ml). After the salts had crystallized, the slurry was cooled to room temperature, stirred for 12 hours and then cooled to 4 °C and stirred for a further 12 hours. The product was then filtered off, washed with cold ethyl acetate/hexane (2x5 ml) and cold hexane (2x5 ml) and dried in vacuo to give 8 g of the desired product.

In a dry 50 ml flask under an argon atmosphere, • ·

260

product prepared in step (b) (5.0 g). Then N,N'-dimethylpropyleneurea (15 ml) is added and the mixture is stirred for 15 minutes. At this point the system is not homogeneous. Then the flask is placed in a water bath at 22 °C and benzyl bromide (2.1 ml, 1.96 equivalents) is added rapidly (no exothermic reaction). Then tert-butylamine (0.90 ml, 0.95 equivalents) is added dropwise (the temperature rises to 27.5 °C during the addition, is maintained there for about 1 minute after the addition is complete, and then decreases). When the temperature drops to 25 °C, the water bath is removed and the reaction mixture is stirred overnight (16 hours). After this time HPLC indicates the reaction is complete.

The reaction mixture was then diluted with ethyl acetate (30 mL) and tert-butyl methyl ether (30 mL). This solution was washed three times with 5% citric acid (1 x 30 mL, 2 x 15 mL) and then washed with saturated sodium chloride (1 x 15 mL). The resulting solution was dried over magnesium sulfate, filtered, and concentrated in vacuo to give 5.85 g of the desired product as an orange oil.

d) (ó>- CH ₂ — oO—N /T\ < O )—ch ₂ —o—c—-]

- S—o—ch ₂ —

O

CN—C(CH ₃ ) ₃

In a dry 50 ml flask under argon atmosphere, the product prepared in step (c) (2.5 g), ethyl acetate (35 ml), triethylamine (0.98 ml, 1.6 equivalents) and dimethylaminopyridine (0.16 g, 0.3 equivalents) are charged. The reaction mixture is left overnight (about 20 hours). HPLC analysis indicates 99.4% reaction completion. Then

261 • · the reaction mixture was filtered through a thick glass frit and the solids were washed twice with ethyl acetate. The filtrate was washed with 5% citric acid (2 x 15 ml) and once with saturated sodium chloride (15 ml). The organic phase was dried over magnesium sulfate, filtered and concentrated in vacuo to give 3.46 g of the desired product as a tan foam.

E)

O

A 12 L three-necked round-bottomed flask was charged with 10% palladium on carbon (50.34 g, 47.31 mmol) as catalyst, water (362 mL), ethanol (6883 mL), and the product prepared in step (d) (345 g). The mixture was stirred and bubbled with nitrogen for about 20 to 30 minutes, and then hydrogen was continuously introduced into it at a temperature of 15 to 25 °C until the reaction was complete by HPLC. Nitrogen was then introduced into the reaction mixture for about 20-30 minutes, then the mixture was filtered and the filter was washed with 2 L of ethanol/water (95/5). The ethanol/water was partially removed by concentration in vacuo at room temperature to obtain a solution containing one gram of product in 8 to 10 mL. Concentration gave a cloudy to white solution.

The above solution is allowed to crystallize overnight at room temperature with stirring (120-200 rpm). The product is then filtered off and the filter cake is washed three times.

262 • 9 · · · · • · ·

300 ml of cold ethanol/water (95/5, 0 to 5 °C). The filter cake is then dried under vacuum for 10 to 20 minutes. The solid obtained is then dried to constant weight in a vacuum oven at room temperature to give 155 g of the desired final product as a white solid.

and)

To a stirred solution of diisopropylamine (2.02 mL, 14.39 mmol) in tetrahydrofuran (18 mL) was added dropwise a 2.5 M solution of butyllithium in hexane (5.75 mL, 14.39 mmol) at 0°C. The reaction mixture was stirred for 30 min, then the solution was cooled to -30°C and a solution of (4S)-N-(tert-butyldimethylyl)azetidinonecarboxylic acid (1.50 g, 6.54 mmol) in tetrahydrofuran (8.0 mL) was added dropwise. The reaction mixture was then stirred for 30 min at -20°C to -30°C. A solution of the iodine derivative prepared according to Example 192, step (b), (3.02 g, 7.85 mmol) in tetrahydrofuran (4.0 mL) was then added over 10 min. After 2 hours, the reaction mixture was warmed to 0 °C and quenched with ice-cold water (35 mL). The pH was adjusted to 12.5 with 10% potassium hydrogen sulfate. After stirring for 30 minutes, the solution was washed with hexanes, cooled to 0 °C, and acidified to pH 3.0 with 5 N HCl. The aqueous solution was then saturated with sodium chloride.

263 • « ·♦ ··· · * ··» ··· ·· • · · · * · · · · · • · · · t «··*··· ·

and extracted with ethyl acetate (twice). The organic extracts were then combined, washed with brine, dried over magnesium sulfate, and concentrated to give 1.10 g of the desired product.

To a stirred solution of the product prepared in step (a) (1.10 g, 2.96 mmol) in dimethylformamide (10 mL) was added sodium bicarbonate (0.62 g, 7.40 mmol). Benzyl bromide (1.76 mL, 14.78 mmol) was then added. After 24 hours, the reaction mixture was partitioned between ethyl acetate and water. The organic phase was then separated, washed with brine, dried over magnesium sulfate, and concentrated. The crude product was purified by silica gel chromatography to give 1.26 g of the desired product.

c) (H ₃ C) ₃ -c—0—0

-j?H— CH(CH ₃ ) 2 CH(CH ₃ ) ₂

The benzyl ester product prepared in step (b) (227 mg, 0.491 mmol) and 1-diisopropylmethoxycarbonylpiperazine-4-carbonyl chloride (178 mg, 0.614 mmol) were dissolved in methylene chloride (2.0 mL). Triethylamine (103 μΐ, 0.737 mmol) was then added followed by dimethylaminopyridine (6.0 mg, 0.049 mmol).

264

After 48 hours, the reaction mixture was concentrated and the residue was partitioned between ethyl acetate and water. The organic phase was separated, washed with 5% sodium hydrogen sulfate and saturated sodium chloride, dried over magnesium sulfate and concentrated. The residue was purified by silica gel chromatography to give 300 mg of the desired product.

To a stirred solution of the product prepared in step (c) (300 mg, 0.418 mmol) in methylene chloride (1.8 mL) was added dropwise trifluoroacetic acid at 0 °C. The reaction mixture was then stirred at room temperature. After one hour, the reaction mixture was concentrated in vacuo. The crude product was dissolved in chloroform and concentrated. This procedure was repeated three times. The crude product was then dried in vacuo to give 297 mg of the desired product.

To a stirred solution of the product prepared in step (d) (198 mg, 0.314 mmol) in methylene chloride (1.50 mL) was added

265 • · • · · · · ·* • « · » · · • a · » « · • «»··>·· » acetaldehyde (15.0 mg, 0.330 mmol). Acetic acid (36 μΐ, 0.628 mmol) was then added. After 20 minutes of stirring, sodium triacetoxyhydroborate (100 mg, 0.471 mmol) was added. After 24 hours, the reaction mixture was diluted with ethyl acetate. One drop of acetic acid was then added and the reaction mixture was quenched with water. The organic phase was separated, washed with sodium chloride, dried over magnesium sulfate, and concentrated to give 175 mg of the desired product.

The product prepared in step (e) (174 mg, 0.264 mmol) and N,N'-dicarbobenzyloxy-S-methyl isothiourea (95 mg, 0.264 mmol) were dissolved in dimethylformamide (1.5 mL). Mercuric chloride (72.0 mg, 0.264 mmol) was then added followed by triethylamine (110 μΐ, 0.792 mmol). After 3 hours, the reaction mixture was diluted with ethyl acetate and filtered to remove mercury salts. The filtrate was washed with saturated sodium chloride, dried over magnesium sulfate and concentrated. The crude product was purified by silica gel chromatography to give 45 mg of the desired product.

g) (homochiral)

£O ₂ H v_/ (CH ₃ )

CH(CH ₃ ) ₂

About »· · • ·

266

The product prepared in step (f) (45 mg, 0.053 mmol) was dissolved in 1,4-dioxane (0.5 mL). 1 N HCl (53 μΐ, 0.053 mmol) was then added followed by the catalyst, 10% palladium on carbon (15 mg). A hydrogen atmosphere was introduced via a balloon. After stirring at room temperature for 3 h, the reaction mixture was diluted with water:1,4-dioxane (1:1) and filtered. Lyophilization of the filtrate gave 25 mg of the desired product. (M+H) ⁺ 497.

The above compound was prepared and isolated as a trifluoroacetic acid salt by the method of Example 195 except that benzaldehyde was used in step (e) instead of acetaldehyde. (M+H) ⁺ 559.

Example 197 (homochiral)

• ·

267

and)

To a solution of 6-phenylhexanoic acid (4.0 g, 20.81 mmol) and hydroxybenzotriazole (3.50 g, 22.89 mmol) in anhydrous methylene chloride (100 mL) was added ethyl 3-(dimethylamino)propylcarbodiimide hydrochloride salt (4.39 g, 22.89 mmol) at 0 °C. The resulting mixture was stirred for 30 min. 1-tert-butoxycarbonylpiperazine (3.88 g, 20.81 mmol) and diisopropylethylamine (4.35 mL, 24.97 mmol) were then added, resulting in a homogeneous solution. The solution was slowly warmed to room temperature over 3 h and stirred overnight. The solvent was then replaced with ethyl acetate (300 mL). The resulting solution is then washed with 0.25 M potassium hydrogen sulfate (pH 3-4), saturated sodium bicarbonate (pH 9-10), and brine, dried over magnesium sulfate, and concentrated to give the desired product in crude form as a colorless oil.

The crude product prepared in step (a) was dissolved in methylene chloride (160 ml). The solution was then cooled to 0°C and trifluoroacetic acid (40 ml) was added dropwise. The ice bath was then removed. The mixture was then stirred for one hour at room temperature. The solvents were then removed in vacuo. The residue was then diluted with ethyl acetate (200 ml). The solution was neutralized with saturated sodium bicarbonate (pH 10). The aqueous layer was then extracted with ethyl acetate (2 x 100 ml). The combined ethyl acetate solution was then washed with brine, dried

268 • Β · • · · · · with magnesium sulfate and concentration gives the desired product in the form of a colorless oil.

C)

• · · · • · · • · · · • 4 · ·

A solution of phosgene (20% in toluene, 50 mmol, 26.3 ml) was dissolved in methylene chloride (30 ml) and cooled to 0 °C. Then, a solution of the crude product prepared in step (b) and triethylamine in methylene chloride (30 ml) was slowly added to the above solution over 20 minutes. The resulting solution was stirred for 1.5 hours at 0 °C. The precipitate was filtered off. The filtrate was concentrated and the residue was purified by chromatography on silica gel (hexane:ethyl acetate 2:1, Rf=0.15) to give the desired product as a white solid. (M+H) ⁺ 323.3; IR (KBr) 1731 cm ^-1 .

d)

O

To a solution of the benzyl ester product prepared according to Example 1(c) (150 mg, 0.26 mmol) in methylene chloride (3 mL) were added triethylamine (0.043 mL, 0.31 mmol), the product prepared in step (c) (102 mg, 0.31 mmol) and 4-dimethylaminopyridine (1.6 mg, 0.015 mmol). The resulting solution was stirred for 3 hours and then the solvent was removed. Purification of the residue by chromatography on silica gel (hexane:ethyl acetate 1:1, Rf=0.22) gave 210 mg of the desired product as a colorless oil. (M+H) ⁺ 859.5,

269 • · · · · · 0 • · · (M-H)” 857.5.

E)

NH»HC1 (homochiral)

H ₂

ř (CH ₂ )

A mixture of the product prepared in step (d) (100 mg, 0.115 mmol), palladium on carbon catalyst (10%, 30 mg) and 1 N HCl (115 μΐ, 0.115 mmol) was stirred under hydrogen atmosphere (balloon) for 45 min at room temperature. Analytical HPLC after this time indicated the reaction was complete. The reaction mixture was then diluted with water (6 mL), filtered, and lyophilized to give 53 mg of the desired product as a white powder. (M+H) ⁺ 501.3, (MH)- 499.2; IP (KBr) 1785 cm i .

Example 198

and)

HCl (diastereomeric mixture)

*—C-(CH ₂ ) ₅ 270

A solution of the product prepared according to Example 172(f) (80.5 mg, 0.20 mmol), triethylamine (0.056 mL, 0.40 mmol), the product prepared according to Example 197(c) (97 mg, 0.30 mmol), and dimethylaminopyridine (6 mg, 0.05 mmol) in methylene chloride (1 mL) was stirred under argon for 21 hours at room temperature. The reaction mixture was then concentrated in vacuo and the residue was taken up in ethyl acetate, 10% potassium bisulfate, and water. The ethyl acetate layer was then washed again with dilute potassium bisulfate, water (2 x), and brine, dried over sodium sulfate, and concentrated to an oil (179 mg). Chromatography of the above oil on silica gel using 15% and then 25% ethyl acetate in methylene chloride afforded 122 mg of the desired product as an oil.

By processing the product prepared in step (a) (120 mg,

0.174 mmol) with trifluoroacetic acid in methylene chloride as described in Example 172 step (h) gave 174 mg of the crude desired product.

C)

O-CH ₂ ·

O

271 • ·

Treatment of the crude product prepared in step (b) with N,N'-dicarbobenzyloxy-S-methylisothiourea according to the method of Example 172, step (i), gave 204 mg of crude product. Purification of the product on silica gel using methylene chloride:ethyl acetate (3:1) gave 79 mg of the desired product as an oily residue.

d) (diastereomeric mixture)

O

The product prepared in step (c) (76 mg, 0.085 mmol) was hydrogenated in dioxane (3 mL) and 1.0 N HCl (0.085 mL, 0.085 mmol) for 1 h at 1 atmosphere of hydrogen pressure in the presence of a catalyst, 10% palladium on carbon (24 mg). The mixture was then filtered through aqueous dioxane, and the filtrate was concentrated to remove the dioxane, filtered, and lyophilized to give 42 mg of the desired product as a white solid. IP (KBr) 1784 cm ^-1 ; HPLC indicated a mixture of diastereomers (62:38).

Example 199 (racemate)

272 » ·

To a stirred solution of the protected dimethylazetidinone prepared according to Example 135(f) (55 mg, 0.12 mmol) in tetrahydrofuran (3 mL) was added sodium bis(trimethylsilyl)amide (1 N in hexanes, 0.13 mmol) at -78 °C and the resulting solution was maintained at this temperature for 30 min. A solution of the product prepared according to Example 197(c) (43 mg, 0.13 mmol) in tetrahydrofuran (1 mL) was then added dropwise and the resulting mixture was warmed to -20 °C. After 1 h, the reaction mixture was quenched with saturated aqueous ammonium chloride (5 mL) and extracted with ethyl acetate (2 x 5 mL). The combined organic layers were washed with brine and dried over magnesium sulfate. Filtration, concentration, and purification by column chromatography (silica gel, 50% ethyl acetate in hexanes) afforded 79 mg of the desired product as a colorless oil.

To a stirred solution of the product prepared in step (a) (79 mg, 0.11 mmol) in a mixture of ethanol (1.5 ml), water (0.5 ml) and ethyl acetate (1.5 ml) at room temperature was added

273 palladium on carbon catalyst (10%, wet, 15 mg). The resulting suspension was bubbled with hydrogen for 3 hours and then the reaction mixture was filtered. Concentration and purification by preparative HPLC (YMC ODS A 20 x 250 mm, 5 μ, 0-100% B in 35 minutes, 15 minutes residence time, A=10% methanol in water and 0.1% trifluoroacetic acid, B=90% methanol in water and 0.1% trifluoroacetic acid) gave 42 mg of the desired product as a white solid. (M+H) ⁺ 485.

In a similar manner, using the methods described in Examples 197 to 199, compounds of general formulae I, II, and IV can be prepared, in which Xi or X ₂ have the meanings given below:

example Xi or X ₂

200

O

201

—c N— C NH—(CH ₂ )

202

IÍ / \ ff ~ ^c ~^ /Nc— (ch ₂ ) 4-0-00/

203

204

274 * · · · · • · · · ··· · • ··· ····»·

205 / \ íř .

-l·^ N— C— O-(CH ₂ ) 3

206

207

208

209

210 yr

Ne—NH—(CH ₂ ) 3' ff /-λ íř r<yo)

C—\f— C—N (CH ₂ ) 3-Q-<^) '-' ch ₃ r~\ ír c—Ιξ ^Nr- C— (CH ₂ ) 3

Al

211

275

Claims (38)

PATENT CLAIMS 1. A compound of the general formula (Ό NH II H ₂ N— c—Αχ- A -j oh Ri Xi (II) NH II H ₂ N—C—NH—(CH ₂ ), q Λ -1 and Rl Xi (III) NH II H ₂ N— C —NH— (CH ₂ ) " q -1 q -Rl Xi (IV) NH H H H ₂ NC—NH—(CH ₂ Ιξ- q -1 q. The Rl X ₂ or (V) NH Rl3 H ₂ N— o—NH-(CH ₂ ) f X3 2Ί6 including an inner salt thereof, or a pharmaceutically acceptable salt thereof, or a hydrolyzable ester thereof, or a solvate thereof, wherein: R 1 represents a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, A ₂ -aryl, O II —c—r ₇ i? IJ — CN-|— ¹ Re( _ch2 )o II —c—o—R ₇ (CH ₂ )o ^(CH ₂ ) _v >^ _ • ^r 8 — CN k_(CH ₂ ) _w (CH ₂ )nb ₂ b ₃ o ll — C—CH ₂ —o—Ri0 -SO2-R7 , R ₉ ^r 9 — CNA ₂ -NCR _{4 t} —cNA ₂ -NC-OR ₄ , —C—alkyl—SO ₂ —R ₇ or O II C-aryl—SO ₂ -R ₇ or R 1 represents an alkyl group represents an alkyl group and R 3 represents, with the exception, hydrogen; that R ₂ R ₂ and R 3 both represent hydrogen, or R ₂ represents an alkyl group with the proviso that in this case R 3 represents hydrogen, or R 3 represents an alkyl group with the proviso that in this case R ₂ represents hydrogen; — C-aryl—S0 ₂ —R ₇ - ^c CH ₂ —O—R _X o , O II —SO2-R7 , C—NH S0 ₂ —R7 ír M -cna ₂ -ncr ₄ R ₉ or — CNA ₂ -N- C-OR4 Ai means -(CH 2 )p- _z —(CH ₂ )p— _; ( ^CH 2 )ra \ (CH ₂ ) _£ —N (CH ₂ ) _q — _{{ C} h ₂ ) _r - (CH ₂ ) -HN-CH ₂ -, -HN-(CH ₂ ) ₂ , or -HN-(CH ₂ ) ₆ R 5 and R 6 independently represent a group selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A 2 -cycloalkyl, A 2 -substituted cycloalkyl, aryl, substituted aryl, A 2 -aryl, A2~substituted aryl, aryl-A3~aryl, A-ary1-A3~aryl, • · 278 • · • · · • · · · · • · · * A · · 4 9 9 9 9 9 94 4 4 9 94 9 heteroaryl, A ₂ -heteroaryl, heterocycloalkyl, A2-heterocycloalkyl, aryl-A3-cycloalkyl, A ₂ -aryl-A ₃ -cycloalkyl, aryl-A ₃ -heteroaryl, aryl-A ₃ -heterocycloalkyl, A ₂ -aryl-A ₃ -heteroary1, A ₂ -aryl-A ₃ -heterocycloalkyl, cycloalkyl-A ₃ -aryl , A ₂ -cycloalkyl-A ₃ -aryl, cycloalkyl-A ₃ -cycloalkyl, A ₂ -cycloalkyl-A ₃ -cycloalkyl, cycloalkyl-A ₃ -heteroaryl, A ₂ -cycloalkyl-A ₃ -heteroaryl, cycloalkyl-A ₃ -heteroaryl, cycloalkyl-A ₃ -heterocycloalkyl and A ₂ -cycloalkyl-A ₃ -heterocycloalkyl. Re represents a group from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A ₂ -cycloalkyl, A ₂ -substituted cycloalkyl, aryl, substituted aryl, A ₂ -aryl, A ₂ -substituted aryl, aryl-A ₃ -aryl, A ₂ -aryl-A ₃ -aryl, heteroaryl, A ₂ -heteroaryl, heterocycloalkyl, A ₂ -heterocycloalkyl, aryl-A ₃ -cycloalkyl, A ₂ -aryl-A ₃ -cycloalkyl, aryl-A ₃ -heteroaryl, A ₂ -aryl-A ₃ -heteroaryl, aryl-A ₃ -heterocycloalkyl1, A ₂ -aryl-A ₃ -heterocycloalkyl, carboxy, alkoxycarbonyl, aryloxycarbonyl, ř λ C-N^ Rs. R4 R s alkoxycarbonylamino, aryloxycarbonylamino, arylcarbonylamino, -N(alkyl)(alkyloxycarbonyl), -N-(alkyl)(aryloxycarbonyl), alkylcarbonylamino, -N(alkyl)(alkylcarbonyl) and -N-(alkyl)(arylcarbonyl). m means number 1 to 5 ff Y represents 0, S, N-R4 , N-SO ₂ -R7 , N—0—A3—R7 279 ’ ::.. ...» · « .· : :': :♦·:· · :j r.«· ·· ·· · ** í ff í i? N—C—A ₃ -° R _7j ν— c— o—a ₃ —r ₇ n— c—a ₃ -c—r ₇ ^N “ ^c -N\_N-R ₄ , Nc-ίξ_N~C—R ₇ O II N-Cf~A « H /—\ H _ _/ NC-CH ₂ —o—R ₇ , NCl·^_Ν-c— 0—R ₇ or oo θ li /—\ í? II N- C- _N— C— Cr ₇ R ₇ represents a group from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, aryl, substituted aryl, A2-aryl, A2-substituted aryl, heteroaryl, A2-heteroaryl, heterocycloalkyl, A2-heterocycloalkyl, aryl-A ₃ -ary1, A2~aryl-A ₃ -aryl, aryl-A ₃ -cycloalkyl1, A2-aryl-A ₃ -cycloalkyl, aryl-A ₃ -heteroaryl A2~aryl-A ₃ -heteroaryl, aryl-A ₃ -heterocycloalkyl, A2-aryl-A ₃ -heterocycloalkyl, aryl-A ₃ -substituted aryl, A2~aryl-A ₃ -substituted aryl, aryl-A ₃ -substituted cycloalkyl, A2-aryl-A ₃ -substituted cycloalkyl, cycloalkyl-A ₃ -cycloalky1, A2-cycloalkyl-A ₃ -cycloalkyl, cycloalkyl-A ₃ -ary1, A2~cycloalkyl-A ₃ -aryl, cycloalkyl-A ₃ ~heteroaryl, A2~cycloalkyl-A ₃ -heteroary1, 280 • · • » * · « »· ·» • · * • flfl· • · • · fl·*· * > cycloalkyl-A3~heteroaryl, cycloalkyl-A3-heterocycloalkyl, A2-cycloalkyl-A3-heterocycloalkyl, cycloalkyl-A3~substituted cycloalkyl, A2-cycloalkyl-A3-substituted cycloalkyl, cycloalkyl-A3-substituted aryl, A2-cycloalkyl-A3~ -substituted aryl, substituted cycloalkyl-A3-cycloalkyl, A2-substituted cycloalkyl-A3-cycloalkyl, substituted cycloalkyl-A3-substituted cycloalkyl, A2~substituted cycloalkyl-A3-substituted cycloalkyl, substituted cycloalkyl-A3-aryl, A2-substituted cycloalkyl-A3-aryl, substituted cycloalkyl-A3-heteroaryl, A2~substituted cycloalkyl-A3-heteroaryl, substituted cycloalkyl-A3-heterocycloalkyl , A2~substituted cycloalkyl-A3 ^_ -heterocycloalkyl, substituted cycloalkyl-A3~substituted aryl, A2 ^_ substituted cycloalkyl-A3-substituted aryl, heteroaryl-A3~heteroaryl, A2-heteroaryl-A3-heteroaryl, heteroaryl-A3-cycloalkyl, A2~heteroaryl-A3~cycloalkyl, heteroaryl-A3 ^_ substituted cycloalkyl, A2-heteroaryl-A3~ -substituted cycloalkyl, heteroaryl-A3-aryl, A2-heteroaryl-A3-aryl, heteroaryl-A3-heterocycloalkyl, A2-heteroaryl-A3-heterocycloalkyl, heteroaryl-A3 ^_ substituted aryl, A2-heteroaryl-A3-substituted aryl, heterocycloalkyl-A3-heterocycloalkyl, A2~heterocycloalky1-A3-heterocycloalkyl, heterocycloalkyl-A3-cycloalkyl, A2~heterocycloalkyI-A3-cycloalkyl, heterocycloalkyl-A3-substituted cycloalkyl, A2-heterocycloalkyl-A3-substituted cycloalkyl, heterocycloalkyl-A3 ^_ aryl, A2-heterocycloalkyl-A3 ^- aryl, heterocycloalkyl-A3-substituted aryl, A2-heterocycloalkyl-A3 ^_ substituted aryl, heterocycloalkyl-A3~heteroaryl, A2-heterocycloalkyl-A3-heteroaryl, substituted aryl-A3~ -substituted aryl, A2-substituted aryl-A3-substituted aryl, substituted aryl-A3-cycloalkyl, A2-substituted aryl-A3 ^- cycloalkyl, substituted aryl-A3-substituted cycloalkyl, A2-substituted aryl-A3-substituted cycloalkyl, 281 • · • · substituted aryl1-A3~ary1, A2~substituted aryl-A3~ary1, substituted aryl1-A3 ^_ heteroary1, A2~substituted aryl-A3~ -heteroaryl, substituted aryl1-A3 ^_ heterocycloalkyl, A2-substituted aryl-A3-heterocycloalkyl, or Rs./ A ₂ — r ₄ Rs n and o mean 1 or 2 with the proviso that the sum of n and o is 2 or 3; v and w mean 1, 2, or 3 with the proviso that the sum of v and w is 3, 4, or 5. Re represents a group from the group consisting of hydrogen, halogen, amino, -NH(lower alkyl), -N(lower alkyl)2, nitro, alkyl, substituted alkyl, alkoxy, hydroxy, aryl, substituted aryl, A2-aryl, A2-substituted aryl, aryl-A3-aryl, A2-aryl-A3-aryl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, heteroaryl, A2-heteroaryl, heterocycloalkyl, A2-heterocycloalkyl, aryl-A3-cycloalkyl, A2-aryl-A3-cycloalkyl, aryl-A3-heteroaryl, A2-aryl-A3-heteroaryl, aryl-A3 ^- heterocycloalkyl, and A2-aryl-A3-heterocycloalkyl. B1, B2 and B3 each represent CH, or two of B1, B2 and B3 represent CH and the third represents N, or one of B1, B2 and B3 represents CH and the other two represent N. R9 represents hydrogen or a lower alkyl group. R10 represents a group from the group consisting of alkyl 282 • · A2-substituted cycloalkyl, aryl, substituted aryl, A2-aryl, A2~substituted aryl, aryl-A3-ary1, A2-aryl-A3~ -aryl, heteroaryl, A2~heteroary1, heterocycloalkyl, A2-heterocycloalkyl1, aryl-A3-cycloalkyl, A2-aryl-A3-cycloalkyl, aryl-A3~heteroaryl, A2-aryl-A3-heteroaryl, aryl-A3~heterocycloalkyl and A2-aryl-A3-heterocycloalkyl1. R20 represents a group from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, A2-aryl, and A2-substituted aryl. R21 and R22 independently represent a group selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, A2-cycloalkyl, A2-substituted cycloalkyl, A2-aryl, and A2-substituted aryl. means whole number 2 to 6; means whole number 1 to 6; means number 3 to 5; means 0, 1 or 2; means 1 or 2; means 1, 2, 3, or 4; u means 1, 2 or 3. 283 A ₂ represents an alkylene or substituted alkylene bridge of 1 to 10 carbon atoms, an alkenyl or substituted alkenyl bridge of 2 to 10 carbon atoms containing one or more double bonds, or an alkynyl or substituted alkynyl bridge of 2 to 10 carbon atoms containing one or more triple bonds. A3 represents a bond, an alkylene or substituted alkylene bridge of 1 to 10 carbon atoms, an alkenyl or substituted alkenyl bridge of 2 to 10 carbon atoms containing one or more double bonds, an alkynyl or substituted alkynyl bridge of 2 to 10 carbon atoms containing one or more triple bonds, -(CH ₂ ) _d -O-(CH ₂ ) _e -, -(CH ₂ ) _d -S-(CH ₂ ) _e -, -(CH ₂ )a—O—(CH ₂ ) _e — -(CH ₂ ) a ^- S-—(CH ₂ ) _e — z / -(CH ₂ )a—γ—(CH ₂ )e— -(CH ₂ )a—γ—C—γ—(CH ₂ ) _e — R ₂ i R ₂ i R22 • (ch ₂ ) a—γ—c—γ— (ch ₂ ) _e R21 R22 (CH2)" ^0- — (CH ₂ ) _e — R ₂ i (CH ₂ )—°“ ^{C (CIÍ 2 )e} R21 —( _CH2 )r' R ₂ i (CH ₂ ) _e — íř (CH ₂ ) a—C—(CH ₂ ) e or -γ—cR21 (CH ₂ ) _e — •(ch ₂ ) 284 d and e have a meaning independently selected from the group consisting of zero and an integer from 1 to 6. R 1 represents an alkyl group, and R12 represents an alkyl group; X2 means □(CH ₂ ) _m Ø ₆ (CH ₂ ) _o / vvo II -c— \ A (CH ₂ )n \ ^ ^B 2 ^b 3 (CH ₂ )v^ HC — CN <_(CH ₂ ) _w •Rs •c—CH ₂ -0-Rio '' _____τ, — C-alkyl“SO ₂ -R ₇ -C—NH SO ₂ - ^_ R7 ₎ > O 11 V' T ⁹ n — C— Ν- A ₂ - N- c— OR4 — CNA ₂ -NC-R4 — C-aryl—SO ₂ —R ₇ O ' O n , ¹¹ -CR? . with the proviso, — CR ₇ does not mean a group from the group consisting of alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, and substituted naphthylaminocarbonyl, or means -SO2R7 with the proviso that -SO2R7 does not mean a group from the group consisting of alkylsulfones, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfones, and substituted naphthylsulfones; 285 • · ··· ······ X3 represents a group from the group consisting of phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl and substituted naphthylsulfonyl; and R13 means O -C— (CH ₂ ) v^ About About II II -c— CH ₂ -O-Rio _r —C—alkyl—SO ₂ —R ₇ i? ΪΜ' M — C-aryl—SO ₂ -R ₇ —CN-A2-NC-R4 , •O II year ₉ o II — c- N- A ₂ - N- c— OR ₄ oo 11 11 R? _with the proviso that it does not mean a group from the group consisting of phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, carboxymethylaminocarbonyl, and alkoxycarbonylmethylaminocarbonyl, -SO2R7 with the proviso that -SO2R7 does not mean a group from the group consisting of alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, 286 * · • · naphthylsulfonyl, and substituted naphthylsulfonyl, or ^{0 11} » 'CO R7 _with the proviso that —q—o—R7 has a meaning other than alkoxycarbonyl, or Re with the proviso that when m is 1, 2, or 3, then Re does not represent a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, and aryloxycarbonyl. 2. A compound according to claim 1 of general formula (IV) NH II H ₂ N— C—NH— (CH ₂ ) _q Ri X2 including an inner salt or a pharmaceutically acceptable salt thereof, wherein q is 3; R 1 represents a carboxy group or —C—NH—(substituted alkyl); 287 • · · · · · • · · · · · · -9 · • ··· ······· ff Τ’ T’ ír -c—N—(CH ₂ ) ₂ —Nc—OR 4 , X2 means O II •c\_y or ff Τ’ T’ lf -CN-(CH ₂ ) 2 -NC—R ₄ R6 represents an aminocarbonyl group, —C—NH(alkyl) ) o or ll .,,-,χ -C—N(alkyl) 2 . Y is N-R4, ? ? N— C—A ₃ — R7 N— C—A3- O-R7 íř íř íf íř z“\ II II II vt—p. — Ν N Ra NCOA ₃ -R _7; NO-A3-C— R _{7 ( neb0} \_y R4 in the meaning of Y and X2 represents a group from the group consisting of alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, and heteroaryl; R7 represents a group from the group consisting of alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, aryl, -(CH2)1-4-aryl, -(CH2)1-4-aryl-A3-aryl, ^N X or _{_CH2} —N. ^R5 , Rs; _from Ra -N means amino, NH(alkyl) or -N(alkyl)25 R9 represents a lower alkyl group; A3 means a bond, an alkylene bridge of 1 to 6 atoms 288 • · • · — (CH ₂ ) _d -0-(CH ₂ ) _e — carbon, — (CH ₂ )a—γ (CH ₂ ) _e — or R21 d and e are independently selected from the group consisting of zero and an integer from 1 to 6; and R21 represents hydrogen or a lower alkyl group. 3. A compound according to claim 2, including an inner salt or a pharmaceutically acceptable salt thereof, which is a compound from the group consisting of: _{to CO2H} O /-\ n íT ^nc-(ch ₃ ) ₃ ΎΥ h o O II c-nh ₂ O-CT-CH(CH ₃ ) ₂ CH(CH ₃ ) ₂ 289 • · 0 0 · 0 0 0 0 0 0 0 0 0 0 ^/ \f__ c- o- CH- CH (CH ₃ ) 2 CH(CH ₃ )2 NH And _{Kr H2Nn} 0' SZO ₂ H 1 NV ?y-0-CH ₂ -CH(CH ₃ ) ₂ 0 _k C0 ₂ H - /-Vo-cHí-MCHlcHjHh 0' 290 NH χ H ₂ N Ν' Η £Ο ₂ Η . Ν NCCN-CH— CH(CH ₃ ) ₂ f \_y η ι Ο CH(CH ₃ ) ₂ NH O.χ I ch ₃ ο-C(CH ₃ ) ₃ NH h ₂ n^ ² H ΓΙ' „CO ₂ H N CH ₃ ch ₃ \ II ff CN-(CH ₂ ) ₂ -N—cCH ₃ ch ₃ ch ₃ ch _{3 CO2H} X ^/2\-cO-CH-CH ₂ -CH(CH ₃ ) ₂ II about _CH2 — CH( _CH3 ) ₂ 291 • · about CH(CH ₃ ) ₂ NH X h ₂ nn' H II c-nh-ch ₂ -c-nh ₂ N— ¹ o II O—CH—CH(CH ₃ ) ₂ CH(CH ₃ ) ₂ 4. A compound according to claim 3 of the formula O II ^C ^NC-(CH ₃ ) ₃ H including an inner salt or a pharmaceutically acceptable salt thereof. 5. A compound according to claim 3 of the formula 292 NH χ H ₂ N Ν'',_αΡ02Η Π r~\ f~\ including its inner salt or pharmaceutically acceptable salt 6. A compound according to claim 3 of the formula NH X H ₂ NN H or a pharmaceutically acceptable salt thereof 7. A compound according to claim 3 of the formula O X /Ν ^X N— c—O—CH— CH(CH ₃ )2 c \_/ 1 O CH(CH ₃ ) ₂ or a pharmaceutically acceptable salt thereof. 293 • · II /^ i? _r c- _W -e _H ¹ /—\ ··· ··· · · 8. A compound according to claim 3 of the formula O NH X H ₂ NN' H •nh ₂ -O N—' or a pharmaceutically acceptable salt thereof. 9. A compound according to claim 3 of the formula NH h ₂ : ( _CH2 ): or a pharmaceutically acceptable salt thereof. 10. A compound according to claim 1, of the general formula H H Rl About Xi including an inner salt thereof or a pharmaceutically acceptable salt thereof, wherein: R1 represents a carboxy group, J-O -ίO II c294 • · · · · • · · · · · • ···«·♦· · o II or —c—NH—(substituted alkyl); About j_ Xi represents—C—Ν Y ř TT or \_/ -CN-(CH ₂ ) 2—N—C—OR ₄ . ř Τ ⁹ , T ⁹ ř -CN—(CH ₂ ) 2—N—C—R ₄ t represents 2 or 3; u means 1; R6 represents an aminocarbonyl group, O—C—N(alkyl)2. -C—NH (alkyl) or U-Á-As—R? , N-C-A3-0—Ri ø ø i? N—C—O—A ₃ —R ₇ N—C—A ₃ -CR ₇ ír •N ^N—R ₄ R4 in the meaning of Y and X2 represents a group from the group consisting of alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, and heteroaryl; Y is N-R4, or R7 represents a group from the group consisting of alkyl, cycloalkyl, substituted alkyl, substituted cycloalkyl, aryl, -(CH2)1-4-aryl, -(CH2)1-4-aryl- _A3- aryl, / ^R4 _{from r4} Ν _χ or -CH ₂ —N Rs. Rs N _from R4 R 5 represents an amino group, -NH(alkyl), or 295 • · · · · • « · · · • · · » · · · · N(alkyl)2; Rg represents a lower alkyl group; A3 represents a bond, an alkylene bridge of 1 to 6 carbon atoms, () d~“N (CHo ) — v | or —( _{CH 2} ) _d - O-(CH ₂ ) _e — · R21 d and e are independently selected from the group consisting of 0 and an integer from 1 to 6; and R21 represents hydrogen or a lower alkyl group. 11. A compound according to claim 10 including an inner salt or a pharmaceutically acceptable salt thereof of formula '4. >co ₂ h ⁿ k r-\ ff ° _^NCO-Oi— CH(CH ₃ ) ₂ J CH(CH ₃ ) ₂ 12. A compound according to claim 10 including an inner salt thereof or a pharmaceutically acceptable salt thereof of the formula » · · · · • · · · * · • ·«·»··· · • · v · · Ri 296 13. Compound of general formula (I) NH II h ₂ nc—A _r -j O Xi (Π) NH II H ₂ N—C—NH—(CH ₂ ) q q -1 SF Ri Xi (III) NH H ₂ N—C—NH—<CH ₂ )7 q λ -j 4 About •Rl Xl or (IV) NH Η H II H ₂ N— c— NH—(CH ₂ ) q 0* -1 I. -Rl O X ₂ including an inner salt thereof or a pharmaceutically acceptable salt thereof, or a hydrolyzable ester thereof or a solvate thereof, wherein: A 1 , R 1 , R ₂ , R 3 , R 1 , q, and R ₂ have the meaning given in claim 1; 297 ff ff —C-mN-C— P.25 Xi and Χ2 mean «· ·· .·*.ϊ 9 · » · * · • ··· · ' _β « · * · *···· .····» »·Φ« ♦· ·· · R25 is a linking intermediate group attached to a lipophilic group. 14. A compound according to claim 13, wherein R25 represents a group selected from the group consisting of —N—( _CH2 ) _ch3 —O-( _CH2 ) Ho) , Ho) . —O-(CH ₂ ) Ho) , -(CH ₂ 74 , -nh-(ch ₂ )— δ) , — (CH ₂ ) ₆ -(θ) , NH— (CH ₂ ) 15. The compound of claim 14, wherein R25 means — (CH ₂ ) ₅ · « · · « • · · · · ··· ······· 298 16. A pharmaceutical composition suitable for treating and/or preventing conditions in mammals associated with tryptase, thrombin, trypsin, factor Xa, factor VIIa or urokinase plasminogen activator, comprising an effective amount of a compound according to claim 1, including an inner salt thereof or a pharmaceutically acceptable salt thereof, or a hydrolyzable ester thereof, or a solvate thereof, and one or more pharmaceutically acceptable carriers. 17. A pharmaceutical composition suitable for the treatment and/or prevention of asthma and allergic rhinitis, characterized in that it contains an effective amount of a compound according to claim 1, including an inner salt thereof or a pharmaceutically acceptable salt thereof, or a hydrolyzable ester thereof, or a solvate thereof, and one or more pharmaceutically acceptable carriers. 18. A pharmaceutical composition suitable for the treatment and/or prevention of chronic asthma, characterized in that it comprises an effective amount of a compound according to claim 3, including an inner salt thereof or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, wherein said composition is formulated into a preparation suitable for inhalation administration to the bronchioles. 19. A composition according to claim 18, characterized in that the active ingredient has the formula NH X H ₂ NN' H _A CO ₂ H rf , About II CC O NC-( _CH3 ) _3H 299 • · · · · · · ··· · ··· • · · ··· ···· · · · · · · · · · · · · · · · · · · and includes an inner salt thereof or a pharmaceutically acceptable salt thereof. 20. A method of treating and/or preventing a condition in a mammal related to tryptase, thrombin trypsin, factor Xa, factor VIIa, or urokinase plasminogen activator, comprising administering an effective amount of a composition according to claim 16. 21. A method of treating and/or preventing asthma or allergic rhinitis in mammals, comprising administering an effective amount of a composition according to claim 17. 22. A method of treating chronic asthma in mammals, comprising administering an effective amount of a composition according to claim 18 by inhalation into the bronchi. 23. three The method according to claim 22, characterized in that the active ingredient has the formula: O II H O and includes an acceptable salt thereof, an inner salt thereof, or a pharmaceutically acceptable salt thereof 24. A method of treating and/or preventing a medical condition in a mammal that is associated with tryptase, factor Xa, factor VIII, or urokinase, comprising administering an effective amount of a composition comprising 300 • · • · · · compound of formula (VI) NH II H ₂ NC—NH-(CH ₂ ) 7 including a zwitterionic or inner salt thereof, or a pharmaceutically acceptable salt thereof, or a hydrolyzable ester thereof, or a solvate thereof, wherein: f represents an integer from 3 to 5; R14 represents a group from the group consisting of hydrogen, carboxy, alkoxycarbonyl, alkylcarbonyl, phenylcarbonyl, substituted phenylcarbonyl, naphthylcarbonyl, substituted naphthylcarbonyl, alkylsulfonyl, phenylsulfonyl, substituted phenylsulfonyl, naphthylsulfonyl, substituted naphthylsulfonyl, phenylaminocarbonyl, substituted phenylaminocarbonyl, naphthylaminocarbonyl, substituted naphthylaminocarbonyl, _A2 -aryl, or _Όί2 )πι where m represents 1, 2 or 3 and R e represents hydrogen, carboxy, alkoxycarbonyl or aryloxycarbonyl; X 3 has the meaning given in claim 1; and A ₂ represents a straight or branched chain alkylene substituted alkylene bridge of 1 to 7 carbon atoms. 301 • · /X .. ·· · ··! ,.. . · . · % · ···· . ··» · ·χ ,* : ; * ί ι **ϊ* ’ · ·\ • · · * · ·; 25. Compound of the general formula N-Ri II HN—CN—(CH ₂ ) _q kh ,co ₂ zx ₂ where X 2 and q have the meaning given in claim 1; Pi represents an N-protecting group; and Z represents a benzyl or benzhydryl group. 26. Compound of the general formula N-Pi HN—CN—(CH ₂ ) _g · Pl H R13 wherein E13 and q are as defined in claim 1; and Pi represents an N-protecting group. 27. Method for preparing compounds of the general formula NH ll H ₂ N— CN— (CH ₂ ) qH Xi including their inner salts or their pharmaceutically acceptable salts 302 « · • · · · » · · · acceptable salts, where R15 means -C-N R4 Rs. OX(CH ₂ ) _v -x J. ' Y — C—N ^Y □ c—HN—' Rs (CH ₂ ) _o (CH ₂ ) xn -Bi < y — ^c — (CH ₂ )^ (CH ₂ )n ^ ^b 2 b ₃ Rs oi ³ f ³ í? . ø 1 ⁹ f' i? — CN—A ₂ — N— C—R ₄ ^or — CN—A ₂ —N— CR ₄ ' ^and Xi, q, v, w, o, n, Bi, _B2 , _B3 , R6, R7, R8, R9, Ra, R8 and m have the meaning given in claim 1; characterized by including: a) coupling of an intermediate of the general formula CO ₂ H — (CH ₂ ) _q —H N-Pi II HN— CN— (CH ₂ ) _q Pi -NH where Pi represents an N-protecting group, with an amine selected from the group consisting of 303 • · • · · • · « κθ ₂ ) _ν ^ HN ^Y '''-(cn.rí' ···· ·· ·· /* HN^ R ₅ ΗΝ· □ÍCH ₂ ) _m Re / HN^ (CH ₂ ) _O ^zBl. Rs (CH ₂ )n ^ ^B 2 b ₃ T' T ³ ί T'?' í? HN—A ₂ —N—C—R4 or HN—A ₂ —N—C—OR ₄ to form a compound of the general formula N- ^p ! ' ^R 15 HN-CN—(CH ₂ ) _g Pi ^H -NH b) reacting the product prepared in step (a) with an acid chloride selected from the group consisting of ^{0 11} 1 Cl—CN-|—* '(CH ₂ ), Cl— C—R ₇ Re( _CH2 )o II ( I Cl—C—N ^Y Cl—c— •Rs (CH ₂ n/ (CH ₂ )nb ₃ í? ^{0 11} II Cl—C—CH ₂ -O—R10 Cl—C—NH—SO ₂ —R ₇ 304 about Rg R ₉ O ¹ II O II Cl—CN—A ₂ —N—C—R _{4 }} Cl—C—alkyl-SO ₂ —R? > About θ R-9 R9 about Η II II li _C l- _C _ _aryl -SO ₂ -R7 , Cl-CNA ₂ -NC-OR _{4 C1} _ _SQ2 _ _R7 , or reaction with OCN-SO2--R9, to form a compound of the general formula N-Pi HN-CN— (CH ₂ ) _q P _X H *Rl5 N Xi c) processing the product prepared in step (b) to remove the N-protecting groups Pi and obtain the desired compounds. 28. Method for preparing compounds of general formula NH II H ₂ N— CN — (CH ₂ ) qH ,co ₂ h Xi including their inner salts or pharmaceutically acceptable salts, wherein Xi and q have the meaning given in claim 1, characterized in that it comprises: 305 • · a) reaction of a compound of the general formula .CO ₂ H N-Pl II HN—CN—(CH ₂ ) _q IH Pi -NH O where Pi represents an N-protecting group with an alcohol of the general formula HO-Z or with bromide or with iodide of the general formula Br-Z or I-Z, where Z represents a protecting group selected from the group consisting of a benzyl group and a benzhydryl group, to form a compound of the general formula N-Pl II HN—CN—(CH ₂ ) I H Pl,CO ₂ Z —NH b) reacting the product prepared in step (a) with an acid chloride selected from the group consisting of O II ci— c—r ₇ s? CT Cl—CN-|— ¹ Re 306 • · • · · · · • · · · Bl. Rs cl—C—N ^(CH ₂ ) _v i \ <^ ^B2 b ₃ or ₉ r ₉ o I Cl—c—CH ₂ -O— R _1o Cl—C—NH—SO ₂ —R ₇ Cl—CN—A ₂ —N—C—R ₄ O Rg R ₉ O II II li cl- c- N—A ₂ —N— c- OR ₄ oo II II Cl—SO ₂ -R _7j C1—C—alkyl—SO ₂ —R 7 or Cl—C—aryl—SO ₂ —R ₇ or reaction with OCN-SO2-R 7, to form a compound of the general formula N-n HN-ON-(CH ₂ ) _q ,co ₂ z -N Xi where R4, Rβ, R7, R8, _R9 , R10, v, w, o, n, m, B1, _B2 , _B3 , and _A2 have the meaning given in claim 1; c) treating the product prepared in step (b) to remove the protecting group Z and the N-protecting groups Pi to form the desired compounds. 29. A process according to claim 28 for the preparation of a compound of the formula NH χ h ₂ nn' H kCO ₂ H O II -f NCN—C(CH ₃ ) ₃ ií '-' ^H 307 • · • · or its internal salts or its pharmaceutically acceptable salts, characterized in that it comprises: a) reaction of a compound of formula nf ,co ₂ h NH b) reaction of the benzyl ester step (a) with the carbamoyl chloride of the product prepared in formula II NCN—C(CH ₃ ) ₃ H to form an azetidinone of the formula _{CO2CH2 ​} NN— C— N— C(CH ₃ ) ₃ \__/ H 0' 308 • · · · • · c) treating the product prepared in step (b) with hydrogen in the presence of a palladium-on-carbon catalyst to remove the benzyl ester protecting group and the benzyloxycarbonyl N-protecting groups and obtain the desired compound. 30. A compound of the general formula where H ₂ N—CN—(CH ₂ ) _q Pi q Xi have the meaning given in claim 1; and Pi represents an N-protecting group. A compound according to claim 30 wherein: q means 3; Xi means Pi means About f* O II O II NCN— C(CH ₃ ) ₃ ' Compound of the general formula 309 • « • · • · where q is an integer from 1 to 6; and Pi represents an N-protecting group. 33. A compound according to claim 32 wherein q is 3; and Pi means O 34. Method for preparing compounds of the general formula NH H H H ₂ N- C- NH-(CH ₂ )<j- q -th 1 COOH O X1 or their inner salts or their pharmaceutically acceptable salts, wherein q and X1 have the meaning given in claim 1, characterized in that it comprises: a) reaction of azetidinone of the general formula C1-(CH ₂ ) _ _χ ΟΟ ₂ Η J—np ₃ o where P3 represents a silyl protecting group, with 310 with benzyl chloroformate in the presence of triethylamine and dimethylaminopyridine to form a compound of the general formula ·« ·« ·« « • · · · · · «··· · ♦ · * 4 · · » · ····· C1-(CH ₂ ), ₍ /C0 ₂ —ch ₂ — Jnp ₃ b) treating the benzyl ester product prepared in step (a) with sodium iodide to form a compound of the general formula Yes ₃ o c) reacting the iodine derivative prepared in step (b) with a di-protected guanidine of the formula N-Pi II Pl— N-C-NH2 H to form an azetidinone compound of the general formula N-Pi H ₂ N— cn— (CH ₂ ) Pl J—np ₃ o where Pi represents an N-protecting group; d) reacting the azetidinone product prepared in step (c) with ammonium fluoride to form an azetidinone of the general formula 311 ·« *· ·· · • « 0 9 9 9 9 9 99 9 ··< _φ · ··· 9 9 9 9 9 9 9 9 9 9 9 9 99 9 99 9 9 9 N-Px II C-NPi e) reacting the azetidinone product prepared in step (d) with an acid chloride selected from the group consisting of •(CH ₂ ) _m O II Cl—c I- ^v _D cl—C—N cl— c—r ₇ x(CH ₂ )^ l \ro II ci-c Re (ch ₂ )t/ <J μ (CH ₂ )nb ₃ r ₈ Cl—C—CH ₂ -O— R ₁₀ Cl—C—NH—SO ₂ —R ₇ About Rg RgO Cl-CN—A ₂ —N—C—R ₄ Cl—C—alkyl-SO2—R7 *" 9 ⁹ ir ir ir' T ⁹ ir Cl—C—aryl—SO ₂ — ^R 7 Cl— C— N—A ₂ NC OR ₄ , or CI-SO2-R7, or reaction with OCN-SO2-R7, where R ₇ , R6, m, v, w, Y, o, n, Bi, B ₂ , B ₃ , Rs, R9, R10, R4 and A ₂ have the meaning given in claim 1, to form a compound of the general formula N—P II ¹ H ₂ N—CN— (CH ₂ )qPl _Z CO ₂ —CH ₂ ⁰ Xi 312 «9 · ·· ·· • · * f) processing the benzyl ester product prepared in step (e) to remove the benzyl protecting group and the Pi protecting groups and obtain the desired products. 35. A process according to claim 34 for the preparation of a compound of the formula NH at h ₂ n N' H O II _X N NCN—C(CH ₃ ) ₃ £ ^H o or an inner salt thereof or a pharmaceutically acceptable salt thereof, characterized in that it comprises: a) reaction of azetidinone of formula _{kCO 2} H —nJh ₃ Si-C(CH ₃ } ₃ ch ₃ with benzyl chloroformate in the presence of triethylamine and dimethylaminopyridine to form a compound of the formula .C0 ₂ ch ₂ Si—C(CH ₃ ) ₃ ch ₃ • · 313 • · • · • · · • · · · · · b) treating the benzyl ester product prepared in step (a) with sodium iodide to form a compound of the formula c) reacting the iodine derivative prepared in step (b) with a di-protected guanidine of the formula O to form azetidinone of the formula Si—C(CH) ₃ ch ₃ d) reacting the azetidinone product prepared in step (c) with ammonium fluoride to form an azetidinone of the formula O About • · 314 • · · · · · e) treating the azetidinone product prepared in step (d) with a carbamoyl chloride of the formula _About _ _{_} About II /\ II Cl— c— N\ N- C- NC(CH ₃ ) ₃ and obtaining the azetidinone of the formula O About and f) treating the azetidinone product prepared in step (e) with hydrogen in the presence of a palladium-on-carbon catalyst to remove the benzyl protecting group and the benzyloxycarbonyl protecting groups and obtain the desired product. 36. Process for preparing azetidinone compounds of the general formula N—Pi II HN—CN—(CH ₂ )q,CO ₂ H Pl -NH or their amine salts, where Pi represents an N-protecting group and q represents an integer from 1 to 6, characterized in that it includes: 315 a) reaction of a di-protected amine of the general formula P1 ^rl \ z ¹ .NH P1 with an alkyl dihalide of the general formula Cl-(CH ₂ ) q I ; to form a chlorine derivative of the general formula Friday N-(CH ₂ )q ^Cl b) treating the chlorine derivative prepared in step (a) with sodium iodide, optionally in the presence of a base, to form an iodine derivative of the general formula Friday Friday N-(CH ₂ )q -AND c) reacting the iodine derivative prepared in step (b) with a silyl-protected azetidinone of the formula .CO ₂ H Jnp ₃ where P3 represents a silyl protecting group giving, upon removal of the P3 protecting group, an azetidinone of the formula Friday Pi n-(ch ₂ )7 ,co ₂ h NH 316 which can optionally be isolated in the form of its inner salt; and d) treating the azetidinone product prepared in step (c) to remove the Pi protecting groups and reacting the resulting compound with a di-protected guanylating agent of the general formula N—P II Pl-N—C-L H where L represents a leaving group such as methylthio or pyrazolyl, to form the desired compound, which can optionally be isolated as an amine salt. 37. A process according to claim 36 for the preparation of an azetidinone compound of the formula or an amine salt thereof, comprising: a) reaction of a di-protected amine of the formula (h ₃ c) ₃ -c—o—c4nh 2 with an alkyl dihalide of the formula C1-( _CH2 )3-I; 317 »· · • · to form a chlorine derivative of the formula (H ₃ C) ₃ - C— O— C- -N— (CH ₂ ) 3— ^cl b) treating the chlorine derivative prepared in step (a) with sodium iodide in the presence of a base to form an iodine derivative of the formula Oh II (H ₃ C) ₃ -C— o—c N— (CH ₂ ) ₃ —I J2 c) reacting the iodine derivative with the sily1-protected azetidinone prepared in step (b) of the formula xCO ₂ H l si—C(CH ₃ ) ₃ ch ₃ giving, after removal of the tert-butyldimethylsilyl protecting group, an azetidinone of formula (H ₃ c) ₃ -c—o—C--N . 2 which can optionally be isolated in the form of its amine salt; and d) treating the azetidinone product prepared in step (c) with trifluoroacetic acid to remove the tert-butoxycarbonyl protecting groups and reacting the resulting amine with a di-protected guanylating agent of the formula 318 to form the desired compound, which can optionally be isolated as an amine salt. 38. A process according to claim 28 for the preparation of a compound of the formula NH χ H ₂ N Ν' H kCO ₂ H y ncn—C(CH ₃ ) ₃ lf '-' ^H or its internal salts or salts, characterized by a) reacting a compound of formula (I) to make it pharmaceutically acceptable by comprising: 319 with benzyl bromide in the presence of tert.butylamine and N,N'-dimethylpropyleneurea to form the benzyl ester of the formula O (5)-CH ₂ — O- CN í? O)— ch ₂ -ocn ^{/ h} H £O ₂ CH ₂ —O b) reacting the benzyl ester product prepared in step (a) with a carbamoyl chloride of the formula O II Cl—c—N NCN— C(CH ₃ ) ₃ \_/ H for the formation of azetidinone of the formula c) treating the product prepared in step (b) with hydrogen in the presence of a palladium-on-carbon catalyst to remove the benzyl ester and benzyloxycarbonyl groups N-protecting groups and obtaining the desired compound. • · 320 • · ,what ₂ of 39. A compound of the general formula ⁰ Xi wherein Xi, tau have the meaning given in claim 1; P1 represents an N-protecting group; and Z represents a benzyl or benzhydryl protecting group. 40. A method for preparing compounds of the general formula including their inner salts or pharmaceutically acceptable salts, characterized in that it comprises: a) reaction of a compound of the general formula P ₂ —N, .COOH (CH ₂ ) _u -NH O 321 • · where P2 represents a tert-butoxycarbonyl N-protecting group with an alcohol of the general formula HO-Z or with bromide or with iodide of the general formula Br-Z or I-Z where Z represents a benzyl compound of the general formula protecting group, to form P ₂ —N. (CH ₂ ) _u ,CO ₂ Z where tau have the meaning given in claim 1; b) reacting the product prepared in step (a) with an acid chloride selected from the group consisting of (CH ₂ ), Cl—C—R ₇ ⁰ 11 1 Cl—CN- — ¹ Re (CH ₂ ) o ₀ i' 1 l Cl—C—N ^Y (CH ₂ )·^ Cl-C— (CH ₂ )n ^ ^B 2 'b ₃ •r ₈ C!—C—CH ₂ -O— R10 Cl—C—NH—so ₂ —R ₇ About Rg R9 about About Rg RgO I II cl- c- N—A ₂ —N— c—R ₄ cl- c- N—A ₂ —N— C— OR ₄ 322 Cl—SO2-R7 cl—C—alkyl-SO ₂ — ^R 7 or reaction with OCN-SO2-R7, forming the formula C1— C— aryl— S0 ₂ ^R 7 or compounds of general ₀ /-\ Xi where R4, R6, R7, R6, R9, R10, v, w, o, n, m, B1, _B2 , B3 and _A2 have the meaning given in claim 1; c) treating the product prepared in step (b) to remove the protecting group P2; d) reacting the product prepared in step (c) with a di-protected guanylating agent of the general formula N-Pi II Pl-N—C—L H where L represents a leaving group such as methylthio or pyrazolyl to form a compound of the general formula 323 • · · • · · · · · · • · e) processing the product prepared in step (d) to remove the protecting groups Pi and Z and obtain the final desired product.