PL176409B1 - Derivatives of tetrahydropyrane, method of obtaining such derivatives and pharmaceutical composition containing them - Google Patents

Abstract

1. Derivatives of tetra-hydro-pyran with the general formula 1, where R1 refers to the group with the formula 2 or 3, where either R7 refers to a (possibly protected) hydroxyl group, acyloxyl group, a halogen, -OR10, where R10 refers to a lower alkyl, or a (possibly protected) -O(CH2)mOH group, where m is within the range between 2 and 4, or to -OCONH2 group, a R7a refers to hydrogen, or R7 and R7a together refer to an oxo group, R8 refers to a hydroxyl or methoxyl group, while R9 refers to a hydroxyl or acyloxyl group; R2 refers to hydrogen, an acyloxyl group or, possibly, a substituted hydroxyl group, while there is a single or double bond between the two atoms of carbon connected with a dashed line; R3 refers to methyl, ethyl, n-propyl or allyl; or R4 refers to hydrogen or hydroxyl group, while R4a refers to hydrogen, or else R4 and R4a together refer to an oxo group; or else R5 refers to a hydroxyl group, while R5a refers to hydrogen, or else R5 and R5a together refer to an oxo group; and A refers to a group with the formula -CH(OR6)-CH2-(CN2)n- lub -CH=CH-(CH2)n-, where the (CH2)n- part is connected to an atom of carbon, R6 refers to a lower alkyl group, while n is equal to 1 or 2. They are in a free form or, where such forms exist, in the form of salt.

Description

The present invention relates to tetrahydropyran derivatives, their preparation and the pharmaceutical composition containing them. More specifically, the invention relates to macrolide compounds, in particular tetrahydropyran derivatives of the formula I, in which:

Ri is a group of formula 2 or 3, in which either R7 is optionally protected hydroxy, acyloxy, halogen, -OR10 in which R10 is lower alkyl, optionally protected -O (CH2) _m OH, in which m is the number 2 -4, or -OCONH2 and R7a is hydrogen, or R7 and R7a together are oxo, Rs is hydroxy or methoxy and R9 is hydroxy or acyloxy;

R2 is hydrogen, acyloxy, or an optionally substituted hydroxy group, and there is a single or double bond between the two carbon atoms connected by a dashed line;

R3 is methyl, ethyl, n-propyl, or allyl; or R4 is hydrogen or hydroxy and R4a is hydrogen, or R4 and R4a together are oxo; or R5 is hydroxy and R5a is hydrogen, or R5 and R5a together are oxo; and

A is a group of the formula -CH (OR6) -CH2- (CH2) n- or -CH = CH- (CH2) n- in which the (CH2) n- portion is attached to the carbon atom, R6 is a lower alkyl group, and n is the number 1 or 2;

in free form and where such forms exist, in salt form, hereinafter abbreviated as compounds of the invention.

The optionally protected hydroxyl group represented by R2 and R7 in formula I should not be understood to include R2 or R7 otherwise defined as, for example, acyloxy or -OCONH2.

17 (6409

It is further understood that the term protected hydroxyl group or protected -O (CH2) mOH group refers to pharmacologically active compounds having a substituted OH group, a group substantially conferring protection against degradation during chemical synthesis; the protecting group is preferably a conventional hydroxyl protecting group such as tert-butoxycarbonyl or trialkylsilyl, especially tert-butyldimethylsilyl.

Ri is preferably a substituted cyclohexyl moiety as defined above. R2 and R7 preferably represent an optionally protected hydroxyl group, especially a hydroxyl group. R 3 is preferably ethyl or allyl, especially ethyl. R4 preferably together with R4a is oxo. R5 preferably together with Rsa is oxo. Re is preferably methyl or ethyl. Rjj is preferably methoxy. R9 is preferably a hydroxyl group. R10 is preferably methyl, m and n are preferably the number 2. A preferably is -CH = CH- (CH2) n-.

Lower alkyl is preferably straight or branched alkyl having from 1 to 4 carbon atoms, especially 1 or 2, especially 1 carbon atoms. Halogen is fluorine, chlorine, bromine or iodine, preferably chlorine or bromine, especially chlorine. The hydroxyl group is preferably a free hydroxyl group, e.g. unprotected. The acyloxy group is preferably a formyloxy, benzoyloxy or alkylcarbonyloxy group with a total of 2-5 carbon atoms in the alkylcarbonyl portion thereof.

Preferably, there is a single bond between the two carbon atoms connected by a dashed line.

A preferred subgroup of compounds of formula I comprises compounds of formula IV in which the meanings of the substituents are as defined above.

The compounds of the invention have a number of chiral centers and can therefore exist in different stereoisomeric forms. The process of the invention normally produces a mixture of such isomers. Depending on the conditions and type of reaction, the process can be controlled so as to advantageously produce specific isomers. The invention includes all stereo and optical isomers as well as racemic mixtures. The isomers can be separated or separated by traditional techniques. The preferred stereochemistry at the various chiral carbon atoms is represented by Formula 5, wherein the substituents are as defined above.

When R1 is a substituted cyclohexyl group as defined above, wherein R7 and R.7a are not oxo and R7 is not halogen, the preferred stereochemistry is shown in formula 6, while R7 is halogen, it is preferably in the β configuration.

When R5 is hydroxy it is preferably in the a configuration.

Part of OR6 in group A is preferably in the β configuration. This additional chiral center is formed by the process of variant a) for the preparation of compounds of formula

15. The a and β isomers may be separated by a conventional method, eg chromatography.

A further preferred subgroup of compounds of formula 1 are compounds of formula 7 in which:

Rs is a group of formula 8 or 9 wherein either R7S is hydroxy or 2-hydroxyethoxy optionally protected with a tert-butyldimethylsilyl group, or is an isopropylcarbonyloxy group or chlorine, and R7as is hydrogen, or R7s and R7as together represent an oxo group;

R2s is hydrogen, isopropylcarbonyloxy or hydroxy, and a single or double bond exists between the two carbon atoms connected by a dashed line;

R3s is ethyl or allyl;

R4 and R4a and R5 and R5a are as defined above; and As is a group of the formula -CH (OR6s) -CH2- (CH2) n- or -CH = CH- (CH2) n-, where the CH2 group is attached to the carbon atom, R6 is methyl or ethyl and n is given above.

A further subgroup of compounds of formula 1 includes compounds of formula 10 in which R2p and R7p are the same or different and represent an optionally protected hydroxyl group, and the other substituents are as defined above.

176 409

A further subgroup of compounds of formula I comprises compounds of formula 11 in which the substituents are as defined above.

A further subgroup of compounds of formula I comprises compounds of formula 12 wherein

R2 'p is hydrogen or an optionally protected hydroxyl group, and a single or double bond exists between the two carbon atoms connected by a dashed line; or R4p is hydroxy and R4ap is hydrogen, or R4p and R4ap together are oxo;

R7'p is halogen;

and the other substituents are as defined above.

A further subgroup of compounds of formula I comprises compounds of formula 13 in which the substituents are as defined above.

A further subgroup of compounds of formula I comprises compounds of formula 14 in which the substituents are as defined above.

In the subgroup of compounds of formula 14, A is a group of formula -CH = C 1 H 2 - (CH 2) as defined above. In a further subgroup, A represents a group of formula -CH (OR6) -CH2 (CH2) n- as defined above, especially -CH (OCH3) -CH2- (CH2) n-, where n is as defined above.

The compounds of the invention can be prepared by a method which includes:

a) for the preparation of compounds of formula 15 in which the substituents are as defined above, adequate irradiation in the presence of R6OH of the corresponding compound of formula 16 in which the substituents are as defined above, or

b) to prepare compounds of formula 17 in which the substituents are as defined above, an appropriate oxidation of the corresponding compound of formula 18 in which the substituents are as defined above, or

c) to prepare compounds of formula 19 in which the substituents are as defined above, appropriate elimination of R6OH from the corresponding compound of formula 20 in which the substituents are as defined above, or

d) to prepare compounds of formula 21 in which the substituents are as defined above, appropriate reduction of compounds of formula 17; and optionally converting reactive groups in the resulting compounds of formula I having suitable reactive groups, e.g.

- for the preparation of compounds of formula I in which R7 is halogen, the corresponding compounds of formula I in which R7 is hydroxy are halogenated, or

- for the preparation of compounds of formula 1 in which R2, R7 and / or R9 are acyloxy group, the corresponding compounds of formula 1 in which R2, R7 and / or R9 are hydroxy are acylated;

optionally deprotecting one or more of the hydroxyl groups in the resulting compounds of formula I having the protected hydroxyl group (s) ^) or protecting one or more hydroxyl groups in the resulting compounds of formula I having free ( e) hydroxyl group (s);

optionally separating the resulting stereoisomeric mixtures of compounds of formula I into the individual isomers; and recovering compounds of formula I in free form or, where they exist, in salt form.

The process according to the invention can be carried out in a known manner.

The process according to variant a) (irradiation) is a light initiated intermolecular oxidation / reduction reaction. It can be conducted with UV rays and / or visible light, optionally in an inert gas atmosphere, e.g. ultraviolet lamps emitting monochromatic or non-monochromatic UV rays and / or visible light, e.g. at temperatures between about -78 ° C and about 100 ° C C, preferably between about 0 ° C and about 30 ° C, preferably using pyrex filtered light or a long path filter, e.g. an aqueous solution of NaBr and Pb (NOs) 2 in alcohol R6OH or a mixture of alcohol R6OH with saturated or unsaturated hydrocarbon. with an ether, ketone or ester or a mixture of these solvents, which solvents or their components can participate in the reaction or act, for example, as a sensitizer, and optionally in the presence of further

A compound that may be involved in the reaction, e.g., as a sensitizer or an electron transfer aid.

Process according to variant b) (oxidation) may be carried out in a known manner, e.g. by reacting a compound of formula 18 with an oxidizing agent which reacts selectively with a hydroxyl group adjacent to an oxo group, such as Cu (OAc) 2, in an alcohol such as methanol or a cyclic ether such as tetrahydrofuran or water in a mixture of these solvents under an O2 atmosphere or in the presence of some other oxidizing agent, preferably at temperatures between about 0 ° C and about 90 ° C.

The process of variant c) (elimination) may be carried out by reacting a compound of formula 20, e.g. with an acid such as HF, HCl or p-toluenesulfonic acid, preferably in a solvent such as acetonitrile, methanol, tetrahydrofuran, dichloromethane or dimethylformamide or water or mixtures of of these solvents, preferably at temperatures between about -30 ° C and about 100 ° C, or by reaction with a salt such as NH 4 Cl or NH 4 Br, preferably in a solvent such as dimethylformamide, preferably at a temperature from about -30 ° C to about 100 ° C. at about 100 ° C, preferably at low pressure, the removal or introduction of the protecting groups of the hydroxyl group may be carried out in the same step or in separate steps.

Process according to variant d) (reduction) may be carried out, for example, by treating the compound of formula 17 with a reducing agent such as hydrogen sulfide, preferably in a solvent or solvent mixture such as pyridine / dimethylformamide, preferably at temperatures between about -70 ° C and about 10 ° C.

The conversion of the reactive groups can be carried out, for example, for halogenation by reaction with a halogenating agent such as dichlorotriphenyl phosphate in the presence of a base in a suitable solvent, e.g. toluene or tetrahydrofuran, at temperatures e.g. between about 0 ° C and 70 ° C and for acylation, by reaction, e.g. in an inert solvent such as acetonitrile or dichloromethane, with an acyl chloride or acyl anhydride in the presence of a binder such as 4-dimethylaminopyridine or with an acid in the presence of a carbodiimide such as dicyclohexylcarbodiimide.

Removal of protecting groups to remove e.g. tert-butyldimethylsilyl or tert-butyldimethylsilyl or tert -bufoxycarbonyl can be accomplished, e.g., by treatment with hydrofluoric or hydrochloric acid in a solvent such as acetonitrile, methanol or a mixture of methanol and ether. Depending on the chosen reaction conditions (e.g. duration or temperature), the removal can be controlled such that either all or only some of the protecting groups are removed. Partial deprotection is particularly desirable where a specific hydroxyl group is to react in the subsequent reaction.

In order to protect the free hydroxyl group or groups, depending on the reaction conditions selected, the reaction can be controlled such that all or only some of the potentially reactive hydroxyl groups are protected. Suitable protecting groups are conventional hydroxyl protecting groups such as tert-butoxycarbonyl or trialkylsilyl, preferably tert-butyldimethylsilyl.

The separation of the obtained stereoisomeric mixtures can be carried out e.g. by chromatography.

The above variants of the process according to the invention, e.g. a) and b) or a) and c), can also be carried out in a single reaction vessel without isolating the intermediate products.

A compound of the invention in free form may be converted into salt form, where such forms exist, e.g., into an acid addition salt, by the conventional method and vice versa.

The compounds of the invention can be isolated and purified from the reaction mixture in a conventional manner.

Insofar as the preparation of the compounds used as the starting materials is not described here, e.g. in the examples, these compounds are known or can be obtained in a known manner from known compounds.

The following examples illustrate the invention. They are not limiting. All temperatures are given in ° C. In NMR spectra, all shift values

176,409 chemicals are given in ppm; samples were determined by CDCb unless otherwise noted. The stereochemical configuration at different carbon atoms is as for FK 50, unless otherwise stated. All compounds are in free form. The following abbreviations were used:

Ac = acetyl cf = colorless foam db = double bond between two carbon atoms connected by a dashed line depr = protective groups removed iPr = isopropyl m = mixture of stereoisomers with respect to the indicated position mp. = melting point O = oxo group

OtBDMS = tert-butyldimethylsilyloxy group sb = single bond between two carbon atoms connected by a dashed line tBDMS = tert-butyl-dimethyl csilyl B = formula 22 group B '= formula 23 group D = formula 24 group E = formula 25 group F = group G = Formula 27 group J = Formula 28 group

FK 506 = a compound of formula 29 where R3 = CH. ₂ CH = CH ₂

FR 520 = a compound of formula 29 where R3 = C2H5.

Example I: Compound of formula 1 [R1 = B; R2 = OH; Sat; R3 = C2H5; R4 = OH (m); Rs = OH (a); R4a, Rsa = H; A = -CH (R ₆ ) CH2- (CH2) 2-, where OR6 = OCH3 (S)] [Process variant: deprotection]

250 mg of the compound of Example Ia (see below), 10 ml of methanol and 0.15 ml of ether saturated with dry HCl were stirred in an ice bath for 2 hours. The mixture was poured into an aqueous bicarbonate solution and extracted with ethyl acetate. The organic phase was dried over MgSO4 and the solvent was evaporated. Chromatography on silica gel of the residue gave the title compound as a colorless foam (cf).

Example Ia: Compound of Formula 1 [R1 = B ', other substituents - as in Example I; sb] [Variant a) of the process (irradiation)]

Argon-degassed solution of 3.8 g of the compound of formula 16 [R1 = B '; R2 = OH; Sat; R3 = C2H5; Rs = OH (a); Rsa = H; n = 2] in 1200 mL of methanol cooled in an ice bath and irradiated in a well-type reactor for 10 hours using a Hanau TQ-150 lamp and using a pyrex glass filter. Three batches were combined and chromatographed by HPLC on a polygosyl column using cyclohexane / isopropanol (9/1) to afford the title compound as a colorless foam (cf).

176 409

Η

Φ

Ν

Ρ

Ο

Ν

Ο • Η

Ν (ΰ-Η

Ν

Φ • η

Οι

Φ *

Ρ (0

C.

Ο

C.

Ν

Ρ

Ρ

Ο (0

Η

Φ • Η

Ν

Ό (0 γΜ> ι

Ν

Ρ

Οι ο

C (0 ω

• Η (0 • γ-,

Φ • Η β

Ν υ

• Η θ '

Ο rH <0

C.

Example VIII: Compound of formula 1 [R1 = B; R2 = OH; Sat; R3 = C2Hs; R4 + R4a = R5 + R5 _a a = 0; A = -CH = CH- (CH2> 2-] [Variant b) of the process: (oxidation)]

A mixture of 5 g of the compound of Example 9 (see below), 1.9 g of Cu (OAc) 2 · H 2 O and 500 ml of tetrahydrofuran was stirred under reflux and under an O 2 atmosphere for 7 hours. The solvent was removed in vacuo, the residue was fractionated between saturated aqueous bicarbonate and ethyl acetate, the organic phase was separated, dried (MgSO4), filtered, the solvent was removed in vacuo and the residue chromatographed on silica gel (pretreated with 2 wt% aqueous NaHCO3 and oven dried ) using ethyl acetate to give the title compound (foam). The foam was crystallized by dissolving in diethyl ether and adding n-pentane in small portions and scraping the walls with a rod to initiate crystallization (mp 169-172 ° C).

Example IX: Compound of Formula 1 [R f = B; R2 = OH; Sat; R3 = C2H5; R4 = OH (m); R4a = H; R5 + R5a = O; A = -CH = CH- (CH2) 2-] [Process Variant c) (Elimination)]

A mixture of 50 mg of the compound of Example 2 and 100 mg of ammonium chloride in 100 ml of dimethylformamide was reacted for 1.5 hours in a rotary evaporator under vacuum at 78 ° C so that the solvent slowly evaporated. The remaining solvent was removed under high vacuum and the residue was partitioned between brine and ethyl acetate. The organic phase was separated, dried (MgSO4), filtered, the solvent was removed in vacuo and the residue was flash chromatographed on silica gel (pretreated with 2 wt% NaHCO3 and dried in an oven) using ethyl acetate to provide the title compound (foam). This foam was crystallized by dissolving in diethyl ether and adding n-pentane in small portions and scraping the walls with a rod to start crystallization (mp 119-123 ° C).

Analogously as described in Examples VIII and IX, the following compounds of formula I were prepared:

continued on page 12

176 409 continued from page 11

about)

N

After

N

ABOUT

-H

Λί

N

RT • H

N tt)

About No. • n

-P

Rl c

o c

Rl &

yeah

P.

After

> <

H • H

H.

Λ

ABOUT

Rl

Ό

Rl • M> 1 ta o

c rt ra • H

X

R) • O

and)

-r-t c

N

O • H tP

About r-t

R) c

176 409

Example XXXIII: Compound of Formula 1 [R '= B; R2 = OH; Sat; R ₃ = C2H5; R4 = R4a = H; R ₅ + R ₅ a = O; A = -CH = CH- (CHz) ₂ -] [Variant d) of the process (reduction)]

H 2 S gas was bubbled for 10 minutes through a mixture of 200 mg of the compound of Example 8, 0.3 ml of pyridine and 4.8 ml of dimethylformamide. The reaction mixture was stirred at room temperature for 24 hours. H 2 S gas was again bubbled through the mixture for 10 minutes. The mixture was stirred at room temperature for an additional two days period. The mixture was diluted with toluene and the solvents were removed under high vacuum. The residue was partitioned between ethyl acetate and a saturated aqueous sodium bicarbonate solution. The aqueous phase was separated, washed with brine, dried over MgSO, and the solvent was removed in vacuo. The residue was flash chromatographed on silica gel (pretreated with 2% aqueous bicarbonate) with ethyl acetate to provide the title compound (foam).

Spectrum Example:

¹ H-NMR - Spectra

I 5.83 mp, J = 2.6 Hz); 5.29 (br, d, J = 44Hz); 4.32 (d, J = 8.5 Hz); 3.70 (dd, J = 1.2 and 9.5 Hz); 5.13 (s).

X major, major index); 5.63 brr (); ; 3.84 (dd ..

XI (main ι ^ κ ^ Ι); 5.69 0m; t; ; 4.65 ddd, J = 2.0; 3.25 (s).

XVII 4.96 t ', J = 2.8; 4.19 (c, c, J = 3.7 and 8, Hz ;; 3.92 (n) ;, 3.63 (d, J = 6.6 Hz); 3.63 (s).

XVIII (approx. 21 mixing ratio of isomers; jh ^ ynya ^ <^ me): 5.6) 9 ^ 1 = 2.7 Η) ;; 5.2, (d, J = 9.0 Hz); 5.10 (d, J = 1.6 Hz); 4.92 (d, J = 10.1 Hz); 4.46 (dd, J = 5.6 and 1.8 Hz).

XXVII tha 11th mixture of the Shomers): 7.11 (d, 1 = 8); ; 6.89 (d, J = 8.1); ; 3.03-2.96 (m;; 2.54 - 2.46 (m); 1.16 -1.12 (m).

XXVIII cca 11 meezzinim of the Szomers); 7.10 (d, J = 8.6 ;; 6.89 (d, J = ');; 4.7, 4.64 (m ;; 6.90 (sd, J = 9.5, 6.0); 2.57 - 2.48 (m); 1.18 -1.12 (m).

XXX 6.86 (d; J = 8.6H); ; 7.13 (<,, J = 8.5 Hz ..

XXXIIa, a mixture of the Szomers); 7.22 (d, J = ', 5;; 6.82 (d, 1 =', 6 ;; 6.719-3.59 (several multiplets); 6.68 (s); 6.60 (s) 0.90 (broad singlet).

XXXIIb (month: crnina of śzomerów); 7.05 (d, 1 = 8.1); ; 6463 (<,,; 5, ^^ 7th;; 5.62th;; 3.90

- 6.85 (m).

XXXIIc (you have ^ i ^ nn ^ a ϊζο (16ι ^); 7.09 (d, J ^^, 7 ;; 6.52 (d, J ^ i ^, 5 ;; 5.69 you;; 5, 64 you ;; 3.07

- 6.0 (m).

' ³ C-NMR - Spectra Ia 176.8-170.6,136.7, 630 ^ 299.3'12 ^, 5, 97.3, 84.3, 80.1

II (main component): 214.0,173.3,170.7,139.7,131.1, 129.7, 12 ^, 97.0,80.5.75.2.52.1.

III (main index): 214.0, 173.1, 170.3, 139.7, 132.6,130.1,123.9, 96.9.84.2.78.2, 76.9, 75.6 ' 74.9, 76.52, 76.46 '72.5, 71.166.7, 52.2.

IV (main component): 213, Ί 176.6, 170, ', 139.9,165.7, 131,', 129.7,126.3,116.5, 97.0, 84.2 '80.5, 77, 7, 75.2, 52.1.

V (main component): 213.1, 173.2, 169.2, 130.0, 122.8, 99.6.88.3.84.2,

78.5, 77.0, 74.1, 76.5, 71.6, 71.2, 69.6 '59.7, 54.2.

VI (main Idadnik): 213.0,173.0, 170, ', 114), 0, 130.3, 123.6, 99.2.88.4.84.1

79.0 '77.6, 74.2' 76.5, 72.2, 71.2, 69.9, 59.8, 55.8, 54.0.

VII (major isomer): 2 (4.1, 173.4. ΉΟΙ, 14.7, 7.12. ^, 70.7.67, '.

176 409

VIII

IX

XII

XIII

XIV

XV

XVI

XIX

XX

XXI

XXII

Xxiii

Xxiv

Xxv

XXVI

XXIX

XXXI

XXXII XXXIII (main component): 213.9,190.8,167.7,162.9, 139.5,131.9,129.8, 123.9,123.2,

109.7, 98.9, 53.0.

(major component): 212.9, 171.2, 168.8,140.3, 132.4, 130.3, 125.4, 123.1, 106.7,

99.5,

195.2,169.5,167.5,136.5,130.6,129.0, 125.3, 96.5, 79.5, 785.2, 53.85, 49.4.

(main ingredient): 213.6, 195.7, 169.0, 167.3, 138.85, 132.25, 130.3, 123.5, 97.0,

84.2, 63.8, 54.5.

(main component), 212,9,195,4,169,1,167.3, 139.0,135.5,131.2, 129.9.1 ^^^ 1L,

116.7, 96.9, 84.3, 79.4.

214.5.188.1.168.6.164.4.140.2.132.3.129.5, ^^ 2. ^^, 98.7, 88.85, 84.2, 78.0, 76.1,

73.5, 71.7, 69.2, 59.0, 57.0.56.6, 56.3, 55.6.55.3

213.6,188.0,168,1,163.9,139.9,131.6,130.0,123.1, 99.1, 88.6, 84.2, 78.4, 76.1,

73.6, 71.4, 69.1, 59.0, 55.0.

169.8,169.0,138.0,131.7,128.3, 127.7, 123.4,109.0, 98.4, 55.4 (main component), 211.9,171.2,168.9,140.4,135.5,132.4,130.4, 125.3, 122.4,

116.5.106.7, 99.5, 84.1, 78.7, 69.0.

213.4, 170.0,169.0,140.3,132.1,130.6,130.4,122.9,109.3,100.0.84.1.78.9.74.1,

73.5, 71.4, 70.5, 69.4, 59.0.

(major isomer): 212.7, 171.5, 168.9, 140.1, 132.1, 131.7, 125.4, 123.1, 106.6,

99.7, 67.1 (ca 2: 3 mixture of isomers): 214.2, 213.3, 168.2, 167.6, 162.9, 139.6, 138.9, 13 ° C, 1 ° C, 131, 13,129.8, 129.2,123.9,123.23,123.16,121.9,112.1,109.8, 98.9, 97.5 (main component): 213.0,194.5 or 190.9,167.7,162.9,139.6,135.4,131, 9.129.8,

123.9.122.6.116.7.109.7, 98.9, 84.2.

213.7,187.0,167.7,159.0,140.3,132.0,130.4,129.4,122.8,113.2.99.2.84.2.79.0,

76.5, 73.58, 73.55, 71.5.69.1, 59.0, 57.7, 56.5, 56.2, 55.4 (main component): 195.7.168, ^, 162, 8,135.4,131.4,128.3,127.9,123.0,109.5,

99.6, 52.6 (approx 45:40 isomer mixtures): 201.4,199.4,194.9,188.8,168.4,167.5,163.3,

163.1, 148.1, 145.3,139.1,138.2, 133.9, 131.4, 129.9,129.6,129.5,127.7, 124.0,

123.7.123.5.121.8.110.9.110.2, 99.0, 98.1

213.8, 195.4, 169.2, 167.4, 138.7, 123.8, 96.9, 83.4, 79.4, 78.3, 74.7, 73.6, 72.6, 71.5, 67.4

213.9, 190.8,167.7, ~ 163,139.5,123.9,123,2,109.7, ~ 99, 67.3

214.8, 170.9,168.1,140.9,132.6,129.2,123.7,122.3,107.3,98.5.84.2.77.4.76.9,

74.2, 73.6, 70.9, 69.4, 37.4

Compounds of formula I in free form and where salts exist, in pharmaceutically acceptable salt form, here abbreviated as agents of the invention, have pharmacological activity. They are therefore indicated for use as pharmaceuticals. In particular, they have anti-inflammatory, immunosuppressive and anti-proliferative effects.

The anti-inflammatory effect can e.g. be determined by the following test methods:

1. Inhibitory degranulation of mammary cells in vitro (test method as described e.g. in EP 569337):

176 409

The agents of the invention inhibit the degranulation of mammary cells (IC50) in this assay at a concentration of as little as 50 nmol.

2. Oxazolone-induced allergic contact dermatitis (in mice) in vivo (test method such as described e.g. by F. M. Dietrich and R. Hess in Int. Arch. Allergy, 38 (1970), 246-259):

The agents of the invention show an activity (inhibition of inflammation swelling) of up to 50% in this test after a single topical application as a 0.01% solution. Hydrocortisone (1.2%) is inactive under these conditions in this model.

3. DNFB (Pig) allergic contact dermatitis in vivo (test method such as that described e.g. in EP 315978):

The double application of 0.13% of the preparation of the agent according to the invention causes the inhibition of the inflammatory reaction up to 50%.

For example, immunosuppressive and anti-proliferative effects can be determined by the following test methods:

4. Inhibition of the proliferative response of lymphocytes to allogeneic stimulation in an in vitro mixed lymphocyte response (MLR) [test method such as that described e.g. in T. Meo, The MLR in the Mouse, Immunological Methods, L. Lefkovits and B. Pemis, Eds , Academic Press, NY (1997), 227-239]:

Agents of the invention suppress lymphocyte proliferation (IC50) in this assay at concentrations as low as 1 nmol.

5. Inhibition of the proliferation of human keratinocytes in vitro (test method as described e.g. in EP 539326):

The agents of the invention are active in this test at concentrations of as little as 7 µmol with an inhibition of approximately 50%.

6. In vitro macrophilin-12 binding assay [test method described by K. Baumann et al., Tetrahedron Letters 34, (1993), 2295-2298]:

The agents of the invention bind macrophilin with an affinity (IC50) comparable to that of rapamycin and FK 506.

7. IL-2 / IL-2 reporter gene assay / in vitro assay [assay method such as described by G. Baumann et al., Transpl. Proc. 24 / Suppl.2 (1993), 43-48]:

The agents according to the invention show IC 50 in this test in concentrations of only 0.2 mmol.

The preferred agent for the foregoing indications is the compound of Example 8 (5,6-dehydro-ascomycin) whose full chemical name is 17-ethyl-1,44-dihydroxy-12 [2- (4-hydroxy-3-methoxycyclohexyl) -1 -methylvinyl] -23,25-dimethOxy-13,19,21,27-tetramethyl-L1, 28-dioxa-4-azatricyclo- [23.3.1.04 ⁹ ] octacose-5,18-diene-2,3.10, 16-tetraon [1R, 5Z, 9S, 12S- [1E (1R, 3R, 4R)], 13R, 14S, 17R, 18E, 21S, 23S, 24R, 25S, 27R].

For example, it can be determined that in Test 3 above, this compound as a 0.13% formulation has better activity than the corresponding 0.13% formulation of dexamethasone. Thus, this indicates that the compound can be administered to larger mammals, for example humans, by a similar regimen of administration at similar or lower dosages than conventionally employed with dexamethasone for the above uses.

The agents of the invention, in particular the compound of Example 8, show less skin atrophy after topical application compared to potent topical corticosteroids and also less systemic side effects compared to FK 506, as demonstrated e.g. by assessing toxicity in rats after subcutaneous administration.

The agents of the invention are therefore indicated as anti-inflammatory agents and as immunosuppressive and anti-proliferative agents for topical or systemic use for the prevention and treatment of inflammatory and hyperproliferative conditions and conditions requiring immunosuppression, such as:

a) treatment of inflammatory and hyperplastic skin diseases, such as psoriasis, atopic dermatitis, contact dermatitis and further eczema dermatoses, seborrheic dermatitis, lichen planus, pemphigus, bullous pemphigoid, epidermal blistering, vasculitis, erythema, cutaneous eosinophils, lupus erythematosus and acne;

176 499

b) prevention and treatment of allergic diseases such as extrinsic asthma, runny nose, conjunctivitis, atopic eczema, urticaria / angioedema, food / drug allergy and anaphylaxis;

c) prevention and treatment:

- resistance in organ or tissue transplantation, e.g. heart, kidney, liver, bone marrow and skin,

- host-versus-graft reaction, such as following bone marrow transplantation, and autoimmune diseases such as progressive rheumatism, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, severe muscle weakness, type I diabetes and uveitis,

- cutaneous manifestations of immune disorders; and

d) alopecia areata.

Agents can be administered systemically or topically. For use in the above indications, the appropriate dose will of course vary depending, for example, on the particular agent employed, the patient, the mode of administration, and the nature and severity of the condition being treated. However, in general, good results have been obtained with intracorporeal administration at daily doses of from about 0.15 mg / kg to about 1.5 mg / kg of animal body weight. An indicated daily dosage in the larger mammal is in the range from about 10 mg to about 100 mg, conveniently administered, for example, in divided doses up to four times a day or in retard form. When applied topically, satisfactory results have been obtained with topical application in an active substance concentration of about 0.1% to about 3% several times a day, e.g. 3 times a day. Examples of galenic forms are lotions, gels, creams, sprays, and solutions.

The agents of the invention may be administered by any conventional route, in particular enterally, e.g. orally, e.g. in the form of tablets, capsules or topically, e.g. in the form of liquids, gels, creams, aerosols, ophthalmic or nasal solutions for the topical treatment of the skin and membranes. mucous membranes, e.g. of the eyes, respiratory tract, vagina, mouth and nasal cavity.

Pharmaceutical compositions, e.g. for topical application, comprising an agent of the invention in combination with at least one pharmaceutically acceptable carrier or diluent may be prepared in a conventional manner by mixing with a pharmaceutically acceptable carrier or diluent. In single dose form, the dose is, for example, from about 2.5 mg to about 50 mg of the active ingredient.

Topical application includes e.g. skin. Further topical application relates to the eyes for the treatment of immune mediated ocular conditions such as, for example, autoimmune diseases, e.g., uveitis, corneal transplantation, and chronic keratitis; allergic conditions, e.g., vernal conjunctivitis; inflammation and corneal transplantation by topically applying an agent of the invention to the ocular surface in an ophthalmic pharmaceutically acceptable carrier.

An ophthalmic carrier is such that the agent is kept in contact with the surface of the eye for a sufficient period of time to allow the agent to penetrate the cornea and inner regions of the eye, e.g., inner chamber, posterior chamber, vitreous humor, aqueous extracellular fluid, vitreous extracellular fluid, cornea, pupil / ciliary body, lens, choroid, eye / retina, and sclera. The pharmaceutically acceptable ophthalmic carrier can, for example, be an ointment, a vegetable oil or an encapsulating material.

Another form of topical treatment is the application of an agent of the invention to bronchial and alveolar epithelium by inhalation, e.g., an aerosol or powder, e.g. in asthmatic patients.

The agents of the invention inhibit the infiltration into the airways of antigen-induced inflammatory cells following topical administration into the airways by the pulmonary route. Therefore, they are indicated for use in the treatment of respiratory or pulmonary diseases such as asthma. This effect can be demonstrated using the standard

176,409 test methods, e.g. by measuring the effect on allergen-induced pulmonary eosinophilia (in vivo) as described in EP 577544.

Accordingly, it is desirable to use the agents of the invention in the treatment of diseases or conditions responsive to or requiring local treatment of the respiratory tract or the lungs, in particular inflammatory diseases or airway obstructive diseases. They are especially indicated for the treatment of diseases and conditions of the respiratory tract and lungs with or characterized by the penetration of inflammatory cells or other inflammatory events accompanied by inflammatory cells, e.g. eosinophils and / or neutrophils. , accumulation, particularly in the treatment of asthma. They are also indicated for use in the treatment of bronchitis or for the treatment of chronic or acute airway obstruction associated with pneumoconiosis and eosinophilic airway disorders.

For the foregoing purposes, the agents of the invention may be employed in any dosage form suitable for topical administration to the desired site, e.g. by the pulmonary route by inhalation from a suitable dosing device, e.g. in any suitable finely dispersed or finely dispersing form capable of being administered for administration to the desired site. the respiratory tract or the lungs, for example in a finely divided form, or in suspension or in solution in any suitable (e.g. pulmonary administration) solid or liquid carrier medium. When administered in a dry fine form, the agents of the invention may, for example, be used as such, e.g. in micronized form without additional materials, diluted with another suitable finely divided inert solid carrier or diluent, in a coated fine form or in any other suitable form known in the art. in this field, pulmonary administration of a finely divided solid form.

Pulmonary administration can be accomplished with a suitable known inhalation or dry liquid drug delivery system, e.g., an aerosol spray, fluid spray, dry powder inhaler, or the like. Preferably, a dispensing device is used, i.e. capable of delivering a predetermined amount of an agent according to the invention with each actuation. Such devices are known.

Pharmaceutically acceptable diluents or carriers acceptable for topical pulmonary administration include, e.g., dry powder formulations of the active ingredient (e.g., an agent of the invention) in substantially pure form, e.g. as used in the art for dry powder delivery from an inhalation device. Means or devices enabling or facilitating topical application include, in particular, an inhalation device, as well as containers and the like from which the active ingredient can be delivered in a form suitable for topical application. Preferred embodiments will be those that allow topical administration to the respiratory tract or lungs, e.g. by inhalation.

Dosages for an agent of the invention for the treatment of diseases or conditions of the respiratory tract or lungs, e.g. for use in the treatment of an inflammatory or obstructive airways disease, e.g. asthma, e.g. by inhalation, are in the order of 0.1 mg to 10 mg per day, e.g. from about 0.5 mg to about 5 mg, preferably from about 1 mg to about 3 mg per day. The dosing will suitably be delivered from the metered amount delivery system in a series of 1 to 5 actuations for each administration, with administration being 1 to 4 times daily. The dosage for each administration will therefore be from about 0.0025 mg to about 10 mg, more preferably from about 0.125 mg to about 5 mg, e.g. when administered from a metered delivery device, e.g., capable of delivering from about 0.25 mg to about 5 mg. about 3 mg of the agent for each operation.

While the anti-inflammatory and immunosuppressive and anti-proliferative effects are the main effects of the agents of the invention, they also possess some degree of activity for increasing the sensitivity to, or increasing the efficacy of, drug chemotherapy. This activity can be determined by the test method described in EP 360760.

The agents of the invention are therefore indicated for use to reverse various types of resistance, e.g., acquired or congenital, to drug chemotherapy or to increase

176 409 drug therapy susceptibility, e.g. as a means to reduce regular concentrations of a chemotherapeutic dosage, e.g. in the case of anti-cancer or cytostatic drug therapy, as a means of reducing the overall drug toxicity, and especially as a means to reverse or reduce resistance, including acquired or congenital resistance to chemotherapy.

The invention therefore also relates to the use of an agent of the invention as a pharmaceutical, in particular anti-inflammatory, and as an immunosuppressive and anti-proliferative agent; an agent of the invention for use as a pharmaceutical; the use of an agent of the invention in the manufacture of a pharmaceutical composition which comprises admixing with at least one pharmaceutically acceptable carrier or diluent; and a method for preparing a pharmaceutical composition which comprises mixing an agent of the invention together with at least one pharmaceutically acceptable carrier or diluent. It further comprises a pharmaceutical composition comprising an agent of the invention in association with at least one pharmaceutically acceptable carrier or diluent. It further comprises a method of treating inflammatory and hyperproliferative conditions and conditions requiring immunosuppression which comprises administering a therapeutically active amount of an agent of the invention to the patient in the event of such administration.

176 409

176 409

PATTERN 2

MODEL 3

176 409

MODEL 5

176 409

MODEL 8

176 409

176 409

MODEL 13

0CH ₃ óch ₃

5α

176 409

176 409

oh ₃ oh ₃

MODEL 16

0CH ₃ oh ₃

MODEL 17

176 409

OCH ₃ OCH ₃

MODEL 19

176 409

PATTERN 21

176 409

XX hoch ₂

WZdR 24

Ct

H ₃ CO

WZdR 25 tBDMS0 (CH ₂ ) A iPrC (0) 0

H ₃ C0 * /

H ₃ CO

WZdR 26 WZdR 27

176 409

Publishing Department of the UP RP. Circulation of 70 copies

Price PLN 4.00.

Claims (10)

Patent claims CLAIMS 1. Tetrahydropyran derivatives of the formula I wherein R1 is a group of formula 2 or 3, wherein either R7 represents optionally protected hydroxy, acyloxy, halogen, -OR10 wherein R10 represents lower alkyl, optionally protected -O (CH ₂₎ m OH wherein m is 2 -4, or -OCONH ₂ and R7a is hydrogen, or R7 and R7a together are oxo, Rs is hydroxy or methoxy and R9 is hydroxy or acyloxy; R2 is hydrogen, acyloxy, or an optionally substituted hydroxy group, and there is a single or double bond between the two carbon atoms connected by a dashed line; R3 is methyl, ethyl, n-propyl, or allyl; or R4 is hydrogen or hydroxy and R4a is hydrogen, or R4 and R4a together are oxo; or R5 is hydroxy and Rsa is hydrogen, or R5 and R5a together are oxo; and A is a group of the formula -CH (OR $) - CH ₂ - (CN ₂ ) n- or -CH = CH- (Ch ₂ ) "- in which the (CH 2) n- portion is attached to a carbon atom, R6 is lower alkyl, and n is 1 or 2; in pc ^ iit ^^ ci free and where such forms exist, in salt form. 2. A compound according to claim Formula 4, wherein the substituents are as defined in claim 1 1. 3. The compound of claim 1 5, wherein the substituents are as defined in claim 1 of formula 5, 1. 4. The compound of claim 1 1 of formula 7, wherein Rs is a group of formula 8 or 9 in which either R7s is hydroxy or 2-hydroxyethoxy optionally protected with a tert-butyldimethylsilyl group or isopropylcarbonylcksyl or chlorine and R7as is hydrogen, or R7s and R7as taken together are oxo; R2s is hydrogen, iscprcpylcarbonyloxy or hydroxy, and there is a single or double bond between the two carbon atoms connected by a dashed line; R3s is ethyl or allyl; R4 and R4a as well as R5 and Rsa are as defined in the claim 1, and A is a group of the formula -CH (OR6) -CH ₂ - (CH ₂ ) _n - or -CH = CH- (CH ₂ ) n-, with the CH2- group attached to the carbon atom, R6 being lower alkyl, and n is as defined in claim 1. 5. The compound according to p. 1 of Formula 10, wherein R2p and R7p are the same or different and represent an optionally protected hydroxyl group, and the other substituents have the meanings given in claim 1, 1 or formula 11, wherein the substituents are as defined in claim 1 or 1, or in this claim or of formula 12, in which R2'p is hydrogen or an optionally protected hydroxyl group, and a single or double bond exists between the two carbon atoms connected by a dashed line; or R4p is hydroxy and R4ap is hydrogen, or R4p and R4ap together are oxo; R7'p is halogen; and the other substituents are as defined in claim Or in claim 1 or of formula 13 in which the substituents are as defined in claim 1 or 1 or in this claim. 6. Compounds according to claim 1 Of formula 14, wherein R2'p and R 7p are the same or different and represent an optionally protected hydroxy group, and the other substituents are as defined in Claim 7. 1. 176 409 Ί. A compound of Formula 1 in which Ri is the group of formula 22, R2 represents a hydroxyl group and there is a single bond between two carbon atoms connected by a dashed line, R3 is ethyl, R4 is a hydroxyl group in the a or β configuration, R5 is a hydroxyl group in the a configuration, R4a and R5a are hydrogen and A is a group of the formula -CH (OR <,) - CH2- (CH2) 2- where OR> is a methoxy group in the β configuration and the stereochemical configuration at various other asymmetrically substituted carbon atoms is as for formula 29 in wherein R3 = CH2CH = CH2, in free form or, if such forms exist, in salt form. 8. Compound of formula 1 which is 17-ethyl-1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxycldohexyl) -1-methylvinyl) -23,25-dimethoxy-13,19,21,27 11,28dioksa-tetramethyl-4-azatricylo [23.3.1.0 ⁴ '9joktakoza-5,18-diene-2,3,10,16-tetraone [1R, 5Z, 9S, 12S [1E (1R, 3R, 4R) ], 13R.,. 14S, 17R, 18E, 21S, 23S, 24R, 25S, 27R] in free form or, if such forms exist, in salt form. 9. A process for the preparation of compounds of formula 1, wherein R1 is a group of formula 2 or 3, wherein R 7 is optionally protected hydroxy, acyloxy, halogen, -OR 10, wherein R d is lower alkyl, optionally protected -O (CH2) _m OH, wherein m is 2 4, or a -OCONH2 group and R7a is hydrogen, or R7 and R7a are oxo, Rs is hydroxy or methoxy and R9 is hydroxy or acyloxy; R2 is hydrogen, acyloxy, or an optionally substituted hydroxy group, and there is a single or double bond between the two carbon atoms connected by a dashed line; R3 is methyl, ethyl, n-propyl, or allyl; R4 is optionally hydrogen or hydroxy and R4a is hydrogen, or R4 and R4a together are oxo; R5 is optionally hydroxy and R5a is hydrogen, or R5 and R5a together are oxo; and A is a group of the formula -CH (OR6) -CH2- (CH2) n- or -CH = CH- (CH2) n, in which the (CH2) n- portion is attached to the carbon atom, R> is a lower alkyl group, and n is the number 1 or 2; in free form, and where such forms exist, in salt form, characterized in that a) to prepare compounds of formula 15, irradiation in the presence of R> OH is the corresponding compound of formula 16, wherein R1, R2, R3, Rs, R5a, R> and n are as defined above, or b) to prepare a compound of formula 17, oxidize the corresponding compound of formula 18, wherein Ru R2, R3, R5 and R5a and A are as defined above, or c) to prepare compounds of formula 19, eliminating R> OH from the corresponding compound of formula 20, wherein R1, R2, R3, R4, R4a, R5, Rsa, R6 and n are as defined above, or d) to prepare compounds of formula 21, a compound of formula 17 is reduced, wherein R1, R2, R3, R5, R5a, and A are as defined above, and optionally reactive groups in the resulting compounds of formula I having the appropriate groups are converted reactive, e.g. - for the preparation of compounds of formula I in which R7 is halogen, the corresponding compound of formula I in which R7 is a hydroxyl group is halogenated, or - for the preparation of compounds of formula 1 in which R2, R7 and / or R9 are acyloxy, the corresponding compound of formula 1 in which R2, R7 and / or R9 are hydroxy is acylated; Optionally, protect one or more of the hydroxyl groups in the resulting compounds of formula 1 having the protected hydroxyl group (s) ^) or protect one or more of the hydroxyl groups in the resulting compounds of formula 1 having a free ( e) hydroxyl group (s); optionally separating the resulting stereoisomeric mixture of compounds of formula I into the individual isomers; and the compound of formula I is collected in free form or, where present, in salt form. 10. Pharmaceutical composition, characterized in that the active ingredient is a compound of the formula 1 in which Ri is a group of formula 2 or 3 in which R7 is optionally protected hydroxy, acyloxy, halogen, -OR10 in which R10 is lower alkyl, optionally protected -O (CH2) mOH in which m is 2-4 , or -OCONH2 and R7a is hydrogen, or R7 and R7a together are oxo, Rs is hydroxy or methoxy, and R9 is hydroxy or acyloxy; R2 is hydrogen, acyloxy, or an optionally substituted hydroxy group, and there is a single or double bond between the two carbon atoms connected by a dashed line; R3 is methyl, ethyl, n-propyl, or allyl; R4 is optionally hydrogen or hydroxy and R4a is hydrogen, or R4 and R4a together are oxo; R5 is optionally hydroxy and R5a is hydrogen, or R5 and Rsa together are oxo; and A is a group of the formula -CH (OR6) -CH2- (CH2) n- or -CH = CH (CH2) _n- with the (CH2) n- portion attached to the carbon atom, R6 is lower alkyl, and n is the number 1 or 2; in free form or, where such forms exist, in pharmaceutically acceptable salt form.