JPS6368598A - Production of n-(s-3-alkyl-heptanoyl)-d-gamma- glutamyl-glycyl-d-alanine and production intermediate - Google Patents

Abstract

(57) [Abstract] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the following formula: [wherein R is methyl or ethyl, R4 and R5 are each hydrogen, or one of R4 and R5 is hydrogen and the other is (C1- C6) alkyl or (C
6-C8) cycloalkylmethyl] An advantageous method for producing an immunomodulator represented by trans-R
-3-alkyl-4-heptenoic acid (VIb below), 5-
3-methyl-6-heptenoic acid (xb below) and 5-3
- Process for the production of alkylhebutanoic acids and production intermediates, represented by the following absolute stereochemical formula: [wherein R is as defined above]; the latter as intermediates in the synthesis of compounds of formula (1) and the following absolute stereochemical formula: % formula % [wherein either (but not both) of the dashed lines represent a double bond, R is as defined above, and R and R are both hydrogen (nla, or a pharmaceutically acceptable salt thereof), or R and R are each independently (C1-
C6) alkyl, (C6-C,)cycloalkylmethyl, or (mb) where R is methyl if the double bond is in the terminal 6°7-position. Concerning regulators (or seventy precursors).

Optically pure 5-3-methylheptenoic acid (11°R-
CH3) is usually one of the quinine salts from the corresponding racemate.
It is produced with variable yields by several crystallizations at 5°C [Lθvene et al., J. Btol, Chem, vol. 95, 1-2].
4 (1932) and 2-n-butyl-butyric acid on page 18.
It is named 4]. Optically active 3-methylheptanoic acid has since been produced by many other methods [Soai et al., p. 215 (1985)].
; U.S. patent of Ohno et al.! 4,564.62
No. 0 (1986); Mori 2808-281
1 page (1981'); Mukaiya
ma) s4, Tam. Chem, Lett,
) 1981.913-916; Posne
r) et al., vol. 103, pp. 2886-2888 (1981)
; Mukaiyama et al., Bulletin of the Chemical Society of Japan (Bun, Chem
, Soc.

Japan) Vol. 51, pp. 3368-3372 (19
78); Mθyers et al., J. A., Am., Kem.

Tsuku, Vol. 98, pp. 2290-2294 (1976)], but these production methods generally have one or more drawbacks (the maximum possible yield is 50%, and at least 50% waste is required, at least 50% of the by-products are carried to the final stage of the manufacturing process, and the raw acid is inherently impure and contains large amounts of organometallic reagents that are difficult to handle. The currently used asymmetric epoxidation (so-called Sharpless decomposition) has been reported so far [Katsuki et al. (Ka
tsuki) and Sharpless
), p. 5976 (1980); Sharpless et al.
Pure 589-604, 1983): Stereospecific Claisen condensation is also known [Ch.
an) et al., U.S. Pat. No. 4,045,475].

The relatively new field of immunotherapy, and specifically the area of immunomodulation, continues to develop rapidly. A variety of naturally occurring compounds have been discovered. Such compounds include N2-C1-(N2-L-threonyl-L
-lysyl)-L-prolyl]-L-arginine, the tetrapeptide tuftsin. Synthetic f:z pebtibiglycan derivatives, particularly those known as muramyldi and petibiglycan, have received particular attention.

Generally, the formula (
1) Immunomodulators and their usage were published on November 2, 1985.
1 horse was released on the 5th, simultaneously pending POT 6th POT/
It is disclosed in US 85102351. Since this application is not yet publicly available, methods for making such compounds and methods for their use are included in this invention to support the utility of this invention.

The immunostimulatory peptides described above are also described in a number of patent specifications:
No. 024,355; D-alanyl-L
- Tetrapeptides and interpeptides containing glutamyl or L-alanyl-D-glutamyl groups are 198
British Patent No. 2,053,231, issued February 4, 1981 and West German Patent No. 3,024, issued January 8, 1981.
No. 281, respectively; derivatives of the C-terminal amino acids garicine or diaminopimelic acid or N-acyl-alanine to γ-D-glutamyl tributylate are described in West German Patent No. 3, issued January 15, 1981. No. 024,369; Lactoyltetrapeptide 1 consisting of N-lactylalanyl, glutamyl, diaminopimeline and carboxymethylamino components is described in European Patent No. 11283, issued May 23, 1980; Formula (~: C○-NH-CH-RdIAl [wherein P is hydrogen or acyl and R1) is especially hydrogen, lower alkyl, human 90oxymethyl or benzyl, Rc and R1 are respectively hydrogen, carboxy, -C
ONRgR (Rg is aqueous, lower alkyl optionally substituted with hydroxy and Hh is mono- or dicarboxy lower alkyl) and Re is hydrogen or carboxy, provided that one of Hd and Ro is hydrogen; is carboxy or -CONRgRh; Rf is hydrogen, m is 1 to 3, and n is θ to 2
The poly, , 16 ptyto 9 and 70 resin in which carboxy and amine groups are retained are described in US Pat. No. 4.
311,640 and 4.322,341, European Patent Application Nos. 25,482, 50,856, 5
No. 1,812, No. 53,388, No. 55,846 and No. 57,419. Also, the above formula [A]
Peptides similar to noheftide (but forming an R' Ham-based amino acid group) are described by Ives et al., U.S. Pat. No. 4,565,653, European Patent Application No. 157.
Peptide 1, in which R4 forms a heterocyclic amino acid, is described in Yves European Patent Application No. 178,845.

(1982) N2(γ-D-glutamyl)-meso-
2(L),2(D)-diamino-pimelic acid with the formula +Al
compound (n is 1 and Ha is CH3CH(OH)
-CO-, Rb is CH3, each of Ro and Ro is -COOH, and R4 is -CONHCH2COO
It is reported as the minimum structure that can exhibit a characteristic biological reaction.

The alternative compound is known as FK-156.

We now use the above formula m (R-C:Ha, R'-R5
A more stable crystalline form of the immunomodulatory compound N-(S-3-methylheptanoyl)-D-r-glutamyl-glycyl-D-alanine has been discovered.

In addition, we are currently investigating the use of organometallic reagents and the R
- Intermediates with high optical purity, avoiding uneconomical by-products of enantiomers.
We have discovered an effective manufacturing method for (■) above.

In the one-step construction route, the first step is the Sharpless-type decomposition of trans-4-alken-3-ols of the formula: where R is as defined above. This step is particularly carried out with titanium tetraisoproboxylic acid and L-...-
The racemate of formula (IV) is oxidized to the S-enantiomer in the presence of diisopropyl tartrate in a relatively inert solvent, sufficient to maintain the desired unreacted R-ethene thiomer of the absolute stereochemistry [Vl]. tert-butyl hibiloberoxide.

In the second step, the compound with formula (■) is ! JC(Ox-
C3) alkyl] orthoacetate, and the intermediate allyl-enol ether is rearranged in the presence of an acid in a solvent inert to the reaction without isolation to give the absolute stereochemical formula (VIa) (C1~
C3) alkyl)(-3-alkyl-4-heptenoate).

(VIa) X-0R";R'-(Ct-03)a
lkyl (VIb) X-0H (VIc) X-cl! (VIa) Forms alkyl hebutanoic acid. In another method, the ester is first hydrogenated to produce (CI-03)alkyl 5-3-alkyl butanoate and then hydrolyzed to form the formula (It).
of acid is obtained.

Yet another method involves the decomposition of trans-4-alke/-3-ol-Yl:acid mercuric [N9(OA
c) 2] Converted to allyl vinyl ether ■ by the action of ethyl vinyl ether in the presence of a catalyst. Next, by heating allyl vinyl ether, the formula (VI (1
) is rearranged to R-4-heptenal in R-3-alcohol,
Next, this is mixed with, for example, Joneθ reagent (dilute #
chromic anhydride in an inorganic acid) to form the above R
-3-alkyl-4-heptenoic acid is formed.

(■) We also investigated the use of organometallic reagents from readily available S-citronellol and the following absolute stereochemical formula: R
- uneconomical by-products of enantiomers and 1! The intermediates with high optical purity, S-3-methyl-6-heptenoic acid (X(1) below) and 5-3-methylheptanoic acid (II, R-OH3 above), were obtained in good total yield. He also discovered an effective method for manufacturing at low cost.

The first step in this synthesis is the conventional method, in which a conventional silyl protecting group (e.g. t-methyldimethylsilyl group) is used.
with the following absolute stereochemical formula: [wherein R8 is hydrogen (Xlla) or zolyl hydroxyl retaining group (Xl[1)]]. ] It produces compounds of the following ar type. The method used is specifically explained in the following production examples, but when the compound of the above formula (■
R8 is the same method previously applied to R-citronellol in the preparation of the R-enantiomer of t-butyldimethylsilyl). The R-enantiomer not used here is used in the synthesis of proxiphomin [Tapolczay
) Gin, Jiei Kem Tsuku,.

Kem Komun 1985. pp. 143-145].

The present invention particularly relates to intermediate compounds of the formula: % formula % and the following step: fat
(b) the compound is a compound of formula (Xa); (b) the compound is a compound of formula (Xa); In some cases, by hydrolysis with dilute inorganic acids, formula (XI))
oxidizing a compound of formula (Xb) with chromic anhydride in a dilute inorganic acid, which step may be carried out as a separate step or carried out simultaneously with the next step; to form 5-3-methyl-6-heptenoic acid of formula (Xi); and if desired +cLl catalytic hydrogenation of the compound of formula (Xd) to form 5-3-methyl-6-heptenoic acid of formula (II) (R is methyl). To form 3-methylheptanoic acid, a compound of formula (XU) can be combined with formula (Xti) and formula (
11°R-OH3) into an optically active acid.

When R8 is hydrogen, there is an advantage of fewer manufacturing steps, but when R8 is a silyl protecting group,
There is an advantage of ease of purification when the intermediate product is isolated from stage +a+.

A preferred human 90oxy protecting group is trimethylsilyl, p-
t-butylphenethyldimethyl-silyl and t-butyldimethylsilyl. A preferred dilute inorganic acid is H2
It is SO4. The compound of formula (Xla) was synthesized by ozonolysis with methyl sulfide treatment of S-citronellol, and the following step: (el) S-citronellol was reacted with t-butyldimethylsilyl chloride 3, (f) the raw hydroxyl It is preferable to synthesize the compound of formula (X11b) (R8 is t-butyldimethyl-silyl) by ozonolyzing the protected citronellol with methyl sulfide treatment.

The invention also provides the following step: la) The activated form of R-4-heptenoic acid or 5-3-methyl-6-heptenoic acid (e.g. the acid chloride VIC or Xe)
of the formula: [wherein R and R are both benzyl or R6
and R7 are benzyl and the other is (C1-)alkyl or (C6-C8)cycloalkylmethyl] to form an intermediate compound of the above formula (llllb) (R and R corresponds to said R and H); and (b) hydrogenating said intermediate compound in a solvent inert to the reaction in the presence of a hydrogenation catalyst, simultaneously reducing the double bond and forming ( It also relates to an improved process for the preparation of immunomodulators of formula (1), comprising hydrolyzing the group(s).

Finally, the present invention provides immunomodulatory agents and/or compounds (1
) regarding the precursor of the above formula @). Compounds of formula (llla) (R-R-hydrogen) relate to ester hydrolysis of diester compounds of formula (Illb).

As used herein, the expression "reaction-inert solvent" means a solvent that does not interact with the unwinding starting materials, intermediates or products to adversely affect the yield of the desired product.

A preferred group for R is ethyl. Preferred acid catalysts are Lewis acids (eg dry HC, 1% AlCe3). Without limiting the invention, racemic trans-
4-hexen-3-ol or trans-4-hepten-3-ol is preferably obtained by reaction of ethyl Grignard reagent with crotonaldehibi or 2-tentenal, respectively.

The invention is easy to implement. R-) Lance-4-alken-3-ols (7) may be obtained by the method of asymmetric epoxidation, the so-called Sharpless decomposition (reference above). According to this method, racemic undesirable S-
ml of trans-2-hexen-4-ol in the presence of a 1 molar excess of L-1+1-diisopropyl tartrate and 1 molar equivalent of titanium tetraisopropoxide in *Vt,
t-Butylhydroxy is present in a small amount per mole of racemic hexenol (both 0.5 mole, but clearly 1 mole).
(less than mol). The reaction is carried out under anhydrous conditions in a reaction-inert solvent, such as methylene chloride, at low temperatures, generally in the range of -20 to -80°C. Isolate the desired unreactive R-trans-2-alken-4-ol by standard methods such as distillation or chromatography. Low boiling point solvents such as pentenes and ethers are preferred for such chromatography due to the relatively high volatility of the desired product.

The next step is to combine R-trans-4-alken-3-ol (tho) and (C1-0
3)) Condensation with realkyl orthoacetate/Claisen rearrangement. Although this reaction is carried out in a reaction-inert solvent, it is generally carried out at a room temperature (e.g.
It is advisable to carry out the process simply in excess orthoacetate ester at 160°C. When triethyl orthoacetate (boiling point 142°C) is the reagent, it is convenient to use the reflux temperature. This reaction is carried out in the presence of an acid catalyst. Propionic acid, pivalen, 2.4-
Acids such as dinitrate/-/l/, dry HCg and AlCl3 are effective. Preferred catalysts are Lewis acids. The most preferred catalyst is AeG13.

In one method, the resulting ester of formula (Ma) is hydrolyzed by conventional methods to yield the corresponding unsaturated acid of formula (■b).

The method of the invention is also easy to implement. Commercially available S-citronellol OJ) was prepared as described above and as described below in F! As explained in the Examples, it is first converted to one of the optically active starting materials of formula (Xn) by conventional methods. Compound (Xn) is converted stepwise to 8-3-methyl-6-heptanoic acid or 5-3-methylheptanoic acid of the above formulas (X(1) and (n)), respectively, as detailed in the following paragraphs. Transform.

In the first step, for example, in an aprotic solvent inert to the reaction, e.g. or
) carry out the reaction. Although temperature is not a critical factor in phospholane formation, preferred temperatures are within the range of ±25°C, most preferably within the range of ±10°C. aldehy)'(Xu&)
Alternatively, a hydroxy-protected aldehyde (XIlb) is reacted with phosphorane in a similar solvent and within a similar temperature range to form a hydroxy- or hydroxy-protected olefin of formula (XI)) or (Xa), respectively.

The second step, which is only necessary if a hydroxy protecting group is present, is to convert the tertiary alcohol (Xb) into the acid (Xa) by hydrolysis, most conveniently the oxidation of the primary alcohol (Xb) to the acid (Xa).
The protecting groups are removed by the aqueous acidic conditions used in the step. Jeans oxidation is carried out using the so-called Jeans reagent, G.
An aqueous solution of H2CrO4 formed from rO3 and a strong acid is used. Dimensional reagents typically contain excess concentrated H2
It is prepared from SO4 and CrO3 and water in a 1=1 if ratio and first diluted with water to a desired concentration, eg about 3M. generally miscible with water, such as acetone,
Alcohol as a solution in an organic solvent inert to the reaction (
XIla), protected alcohol (Xllb)? :
React with at least 2 molar equivalents of Dirons reagent.

Under these conditions, the silyl ether group is rapidly hydrolyzed to form an alcohol, which is then oxidized to the acid (Xa). Temperature is not critical, for example 0-50°C is usually a satisfactory temperature, with ambient temperature (eg 17-27°C) being most convenient.

Di-4nes oxidation occurs through the primary oxidation of an alcohol to an aldehyde of formula (Xc), which aldehyde is
It is not normally isolated by lens oxidation. If isolation of the intermediate aldehyde is desired, oxidation of the alcohol with a more selective oxidizing agent, such as pyridinium chlorochromate, clearly forms the intermediate aldehyde)' (Xc), which can then be Oxidation (by Diene's reagent or other predominant reagent) to form acid (Xd).

If desired, an unsaturated acid (■l)) or (Xa)
in activated form [for example, formula ("4c") or (X13
) as the acid chloride of the formula: [where RIR is independently represents (C1-C6) alkyl, (C, -C8) cycloalkylmethyl or benzyl] and is combined with a compound represented by the formula (m'
forming the immunomodulatory diester compound of b).

This is hydrolyzed in conventional manner to form an immunomodulatory diacid compound of formula ([[la) or a pharmaceutically acceptable salt thereof.

Alternatively, compounds of formula (II t:+ ) (R and/or R or Hensyl) could be hydrogenated to form immunomodulatory compounds of formula (1). In this hydrogenation, the square bond is saturated with hydrogen and at the same time the benzyl group is hydrogenolyzed. Hydrogenation can be carried out with supported (e.g. Raney nickel, Pa/C) or unsupported (e.g. FthC)
J3) is carried out in a solvent inert to the reaction over a hydrogenation catalyst such as nickel or a noble metal. Solvent, temperature and pressure are not critical. Suitable solvents include, but are not limited to, lower alcohols; ethers such as tetrahydrofuran or dimethoxyethane; and esters such as ethyl acetate. To avoid the expense of heating or cooling, it is preferred to use ambient temperature without cooling, even if the reaction is somewhat exothermic. Although the pressure is not critical, it is preferably less than 7 atmospheres to avoid the use of expensive high pressure equipment. Hydrogenation over paZc at a pressure of 3 to 5 times atmospheric pressure is particularly well suited for the reaction according to the invention.

Alternatively, the unsaturated acids of formula (■b) or (Xa) can be hydrogenated under the same conditions as Hakunaka described in the above-mentioned Noragraph, and the 5-3-alkylhebutane a! form (■);
That is, by reversing the hydrogenation/hydrocracking steps K, the 5-3-alkylhebutane e(n> is (C1-C3
) 7/l/Kyl-try-R-3-°alkyl-4-heptenoate (■a) via (C1-C3)alkyl-5-3-alkyl-heptenoate. Finally, the 5-3-alkylheptanoic acid is activated in the manner detailed above, combined with the diester of formula (■) above, and hydrogenated in the manner described above to obtain the immuno- and regulatory compound of formula (1). Form.

In yet another process of the invention, R-)lans-4-alken-3-ol is reacted by the action of ethyl vinyl ether (ethyl It is preferred that the vinyl ether is in excess), at a temperature in the range of 25 to 40 °C (preferably ethyl vinyl ether C boiling point 36 °C)
is converted to the corresponding vinyl ether of formula (DC) at a reflux temperature of .

The vinyl ether (K) produced is generally heated in a high-boiling, reaction-inert oil solvent, such as xylene or decalin, at 140-200°C, if necessary under pressure, to give a steric effect. By specifically translocating
An unsaturated aldehyde of formula (■d) is formed. Formula (fVl
)) to obtain the unsaturated acid of this aldehyde l-#(■
d) is conveniently easily oxidized by the so-called Jeannes reagent (an aqueous H 2 CrO 4 solution formed from CrO 2 and a strong acid). Jinns reagent typically contains excess sulfuric acid and water in a l:1 ff+, quantitative ratio of GrO3.
and diluted with water to the desired concentration (eg, about 3M). To form the unsaturated acid (■b), at least 1 mol of the unsaturated aldehyde (■a)'B is generally added as a solution in a water-miscible, reaction-inert solvent, such as acetone. React with an equivalent amount of Dixons reagent. The temperature is not critical (eg 0-50°C is usually sufficient, but ambient temperature # (eg 17-27°C) is most convenient).

The crystalline form of the compound of formula (1) (R is methyl, R4 and R5 are hydrogen) is obtained by crystallization from an organic solvent or a combination of organic solvents. Suitable solvents are acetone, acetonitrile/ethanol and tetrahydrofuran/ether. The preferred solvent from the standpoint of product recovery is acetonitrile/ethanol, but from the standpoint of product yield, acetone is preferred. This new form has distinct stability advantages over prior art amorphous lyophilizates. This new form is easier to handle and more concentrated.

It is less static, allowing the production of more complex dosage forms.

Pharmaceutically acceptable monobasic salts or monobasic salts of compounds of formula (1) or Cl1l&) are generally prepared by preparing a solution, preferably an aqueous solution of an acid, in appropriate stoichiometric proportions of NaOH,
KO) (obtained by treatment with a base such as Na2CO3 or an amine. The salt is isolated by evaporation or precipitation.

The products of the invention of formula (1) or Qll) are useful as agents for the clinical and therapeutic treatment of diseases caused by various pathogenic bacteria, especially Gram-negative bacteria, in mammals, including humans. These products are also useful as immunostimulants in animals, including humans, at increased risk of infection due to pre-existing or clinically induced immunosuppression.

The test involved the Charles River Breeding Laboratory.
Normal or immunocompromised C from Laboratory)
Use 3H/HθN male mice at °C. Mice were allowed to acclimate for 5 days before use, and then incubated with a divine dilution of the 0.2 at needle (1
The animals were treated subcutaneously (SC) or orally (PO) with test compound (00°1O21 and 0.1μ/kg) or Glacebo (pyrogen-free saline). Treatment regimens depended on the infecting organism used. That is, in normal mice, K'1ebsiela pneumonia
a) 24 and 0 hours before testing against K; and in immunocompromised mice, treatments were performed 3, 2 and 1 days before testing against E. coli or S. aureus. The test is intramuscular administration (IM) in the lower back for K. pneumoniae.
In the case of E. coli and P. aureus, administration was performed by intraperitoneal injection (IP). An amount of 0.2 au was used in the test. Mortality was recorded after 7 days in the case of K. pneumoniae and after 3 days in the case of the other two bacteria tests.

Culture Preparation: Cultures were streaked onto Brain Heart Injection (BHI) agar for purification from frozen blood stocks. Three colonies were picked from the 18 hour plate culture and placed in BHI broth 9. This bouillon culture was grown on a rotary shaker and plated for 3 days.
After 2 hours of growth at 7°C, 0.2 tj was streaked onto several BHI agar plates. 3
After 18 hours of ink-baking at 7°C, slants were inoculated with fc# and Svectronic (sp) with BH broth.
The culture density was adjusted using electronic) 20 and diluted appropriately to reach the LD90 test level in normal mice.

When the compound (I) or (2) of the present invention is used as an infection preventive agent or an immunostimulant in humans, -ff
As standard compositions in trap formulations, they are conveniently administered by oral, subcutaneous, intramuscular, intravenous or intraperitoneal routes. For example, these compounds can be administered as tablets, pills, powders, or granules containing excipients such as starch, lactose, certain clays, and the like. They can also be administered as capsules, mixed with the same or similar excipients. The compounds can also be administered as oral suspension solutions, emulsions, syrups and elixirs with flavoring and coloring agents.

For shed administration of the therapeutic agents of the invention, tablets or capsules containing from about 50 to about soom9 of the active ingredient are suitable for most uses.

The most suitable dosage for each patient will be determined by the physician and will vary depending on the particular age, weight and response as well as the route of administration. Preferred oral dosage ranges are from about 1.0 to about 3001 nq/kg/day in single or divided doses. Preferred parenteral dosage values are from about 10 to about 100 /
L9/day, with a more preferred range being about 1.0 to about 2 oytq/day.

The following examples are set forth to illustrate the invention, but should not be construed as limiting the invention (although many variations of the invention are possible within the scope and spirit of the invention). be.

Example 1 In a 500WLj-3 neck round bottom flask equipped with a magnetic stirrer, septum, thermometer and N2 inlet.

CH, C/2270a/ and 7.2 g of 4A type molequinyl sieves were charged. Ti(OGH(CH3)2')
4 (10,7ate.

0.036 mol) was added via syringe and the mixture was heated to -66°C.
at this point to reduce the viscosity with CH2Cl2
L-m-diisopropyl tartrate diluted with 10-
,1'9. 01043 mol) was added through the cannula, and an additional 5 d of CH2Cl2 was added while washing the cannula. The temperature was raised to -62°C, at which point racemic trans-2-hexen-2-ol (3,609,0
,036 mol) was added and CH2Cl25- was added while washing the cannula.

After stirring for 8 minutes in an acetone/dry ice bath, the temperature was reduced to -68°C, at which point t-butylhydroperoxy) (3M) as a solution in toluene was 7.18ζe0
．． 22 mol) was added via syringe.

The reaction mixture was warmed to -35°C and kept at this temperature in a refrigerator for 1 hour.
It was maintained for 8 hours. The cold mixture was then filtered through a slotted paper and 540 d of acetone and 20° C.
into the stirred mixture, which was cooled to . The mixture was gradually warmed to room temperature and stirred for 20 hours. The mixture was filtered over diatomaceous and washed with CH2Ce2. The combined furnace liquid and washing liquid were stripped without heating to obtain an oily substance (17,629). Since the desired product is volatile in nature, excessive sl-IJ pulsing is avoided. The oil was applied to 179 g of silica gel with xane as eluent:
Chromatographic analysis was performed using inpropyl ether (4:1) with monitoring by TLC (thin layer chromatography). The solvent was distilled off using a short glass helix packed column with a variable take-off head. This removes the solvent with a boiling point up to the distillate temperature of 69.5° C. (from which temperature the distillate temperature begins to decrease) to yield a substantially pure title product as a bottom;
tlcRf 0.25 (hexane:ether 3:1
). A repeat of this experiment used the highly volatile pentane:ether (4:1) as the eluent to facilitate solvent removal and minimize product loss during stripping.

Example 2 The product of the previous example (o, 49), chenyl orthoacetate (3 ml) and pivalic acid were heated in rapid flow for 2.5 hours,
After cooling, the entire reaction mixture was chromatographed on silica gel, starting with hexane:ether (
6:1) to give 0.459 of the product, then eluting with hexane:ether (7:1) to give the purified title product 0.3
59 (C).

Engineering R (film): 2960.1736, 1456, 1367.1334° 1280.1232, 1172, 1028, 962° 8
40 provisions.

Instead of this, pure S-trans-2-hexene-4-
The pentane/ether column containing ether was directly introduced into this stage, and the pentane and ether were first distilled off from the reaction mixture.

Substantially the same results were obtained using propionic acid in place of pivalic acid. Although dried HCe and 2,4-dinitrophenol have been used successfully as acid catalysts, AeC;113, which is used in the following examples, is the preferred catalyst.

Example 3 R-3-Methyl-4-heptenoic acid Method A A 35 al.
-7 Lasconi, implementation full 2 (7) raw meat (1
, 59,0,015 mol). Add chenyl orthoacetate (9m, 0.049 mol) and then
IC:e3 (C,12L O,009 mol) Processed, the mixture was reconstituted on a Soxhlet extractor for 2 hours. By this time, tic has become the intermediate ethyl R-3-methyl-4
- indicated that the conversion to hexanoate was complete; t
/cRf O, 75 (4:l hexane:ethyl acetate),
Rf O, 85 (15:5:2 to #san:ether:
acetic acid).

The reaction mixture was cooled and diluted with 15 m6 of 2 N NaOH and CH
30H12- and incubated for 27 hours at ambient temperature. By this time tdc indicated that hydrolysis was complete. The reaction mixture was stripped of methanol, diluted with 12 ml of H2O, and aH2c12 (
25+1! tX 3 ). - Moronishiri CH2
012 was backwashed with 2N NaOH (25adX 1 ).

Combine the aqueous layer and backwash liquid and use concentrated hydrochloric acid to...IKtJ
Node t,,0H2CJ2'''C-Extracted 3 times, Ta.CH
2Cl2M were combined, dried over JSO4 and stripped to give the title product as an oil (1,289,60%
); lc Rf 0.65 (15:5:2 hexane:ether:acetic acid); 1H-n m r (CD Cg 3) delta (-) 9
．． 4 (8, IH).

-CO2H), 5.5 (m, 2H, -CI-
CH-)t 3.0-1.6 (m, 5H), 1.3-
0.8 (m, 6H); IR (74 m) 3400-24
00,2960,2925,2860°1708.14
58,1410,1380,1295°1228.11
90,1152,1100.930cm.

Method B The ester product of Example 2 was treated with CH30)I25□e
and lN NaOH11,5:πl were added. The mixture was stirred at ambient temperature for 3.5 hours. At this point t
lc indicated that hydrolysis was complete. The mixture was extracted with ether (35++!/x2) and concentrated H (12
and extracted with ether (35aex3). The acid extracts were combined, dried and stripped to give the title product (o,2o!J) which is the same as the product to the process.

Example 4 Ethyl 5-3-methylheptanoate Product of Example 2 (C,2i)'& Paar
Under 4 atmospheres of hydrogen in a hydrogenator, tW(l ethyl 40I
Hydrogenate over 5% Pd/G O, 29 (moistened with 50% water) in ILt for 3 hours. The catalyst is recovered by filtration over cane earth. The title product is recovered by stripping of Solvent F.

Example 5 Method A 5% of the product of Example 3 (C, 2051) in ethyl acid 40 - 1 port under 4 atmospheres of hydrogen in a nord hydrogenation apparatus
Pa 100.21) (moistened with 50% water) for 3 hours. The catalyst was recovered by filtration over kerosene and the oil (C,
The title product was recovered as 29).

When desired, the purified title product was obtained by distillation under high vacuum; bp 77-79°C, 10.2 mm; 1H-nmR.
CDGe3) Deniri (llllff): 12.0
(s, -COOH), 1.0 (d.

-CH5), 0.6-2.8 (m, residual 13H);
IR (Film) 3400-2400, 2960, 29
25゜2B60, 170B, 1458, 1410, 13
80°1295.1228,1190,1152,11
00°930m-1: (α:11) =-6,4
1° (1% in -OH30H); 22.5 n D = 1.427 - Method B The product of Example 4 is hydrolyzed according to Example 3, Method BK to give the title product.

Example 6 Acid product from Example 5 or Example 27 (8.5 g,
0.062 mol) CH2Gg218m/KffJ
I understand. Oxalyl chloride (5.36m, 7.8 OL
0.0614 mol) was mixed into this solution and the mixture was
I left it for a while. During this time the reaction was complete, as evidenced by no more gas evolution. This solution of acid chloride 9 was immediately used directly in Example 8, Method C. Alternatively, the acid chloride was isolated by stripping of the solvent and further purified (boiling point 45°/1,5 order) using method AK of Example 8, if desired by distillation.

Example 7 The acid product of Example 3 (C, 7479.5 mmol) is converted by the method of Example 6 to a solution of the title product in CH2Cl2, which is used directly in Example 9 below. Alternatively, the reaction mixture could be stripped to give the title product, ? 4. If desired, distill under reduced pressure.

Example 8 Method A D-gamma-glutamyl (alpha benzyl ester)-glycyl-D-alanine benzyl ester hydrochloride in methylene chloride 50g (Preparation Example A) 1. o 9 (2,03 mmol) and triethylamine 616 (6,09 mmol), 5-3-methylheptanoyl chloride)'
660 m9 (4.06 mmol) were added and the reaction mixture was stirred at room temperature for 80 hours. The methylene chloride was evaporated in vacuo and the residue was dissolved in ethyl 11ate. The resulting solution was washed successively with 2.5% hydrochloric acid, water, 10% potassium carbonate, water, and brine. The organic phase was separated, dried over magnesium sulfate and concentrated in vacuo. The residue was triturated with diethyl ether and filtered under a nitrogen atmosphere to give the title product, all of which was summarized in Example 10. Method AK was used directly.

Method B Product of Preparation 4 (C,7i, 1.53 mmol), C
H20g25g and triethylamine (C,212rnl
.

1.531 mol) were combined and stirred under N2 atmosphere. 5-3-Methylheptanoic acid (Example 5; 0.209.1.39 mmol) in 24 ml of CH2CI was added followed by dicyclohexylcarbodiimibi (C,
2869° (1.37 mmol) was added and the mixture was stirred for 16 hours. The reaction mixture was filtered, the p-liquid was stripped and the remaining i'k ff1=ethyl acid 1 OILe and the solution was diluted with 2.5% HGe 5mt N205
rslt 10% and 2CO35! Wash sequentially with ILl and saline 5d, M9So.

Drying on top gave the title product 71■ (88%).

Method C In a 500 tptl-four round bottom flask equipped with a stirrer, a thermometer insertion funnel and a N2 inlet, the product of Preparation Example 4 (32,8!l), 05059 mol) was added to a C
H2Cl2175rILl! and cooled to 0-5°C.

While maintaining this temperature range, triethylamine (24
, 7rxl, 17.99. 0.177 mol, 3 eq.) was slowly fed over 15 minutes. Maintaining an ice-water bath, the 5-3- in CH2C71!2 from Example 6
The entire batch of methylheptanoyl chloride 1 was added over 15 minutes while increasing the temperature to 21°C. Stirring in the ice-water bath was continued for 45 minutes, during which time the gelatinous mixture became thick and could no longer be stirred.

The soft gelatin mass was broken and mixed with 10% H (4125 rttl and OH2Cg250 a). The organic layer was separated and 10% H20 (125 door g x 2), 10% 2CO3 (
125ml x 2) Oyohi H20 (125ml 1x
1) Y: Continuous Vcflc cleaning, drying over MgSO4, stripping to a wet white solid 82.
I got 39. These solids were taken up in hot ethyl acetate 500R1. Upon gradual cooling to ambient temperature, the title product precipitated out profusely.

The mixture was further diluted with 40-40 ml of ethyl acetate to maintain easy stirring. The purified title product was collected by filtration and dried under vacuum at 40°C (31.1%
, 90.5%).

"H-nmRCDCe3) delta (P): 8.
4-8.1 (m.

3H), 7.15 (θ, 10H), 5.1 (s, 4H)
, 4.4-4.2 (m, 2H), 3.7 (a, 2H)
, 2. z(t, 2H).

2.1-1.7 (m, 6H), 1.4-1.1 (m, l
0H).

0.92-0.8 (m, 6) 1).

Example 9 N-(R-3-methyl-4-hebutenoyl)-D-γ-
The product of Preparation Example 4 (2,779.5 mmol) is combined with the entire batch of 0820 g2 medium acid chloride from Example 7 according to method C of Example 8. The product is recovered by stripping the washed and dried organic layer, dissolving the residue again in ethyl acetate and restripping.

Example 1O N-(S-3-Methylheptanoyl)-D-[gamma] - Method A The entire product of Example 8, Method A was dissolved in 65 rat of methanol. Palladium hydroxide was added to this ff[, and the mixture was photographed for 3 hours in a hydrogen atmosphere of 4 atm. The catalyst was purified and the solvent was removed under vacuum. Put the residue in water
f4 was thawed and freeze-dried to obtain the desired product.

NMR scan (DMSO-a6) showed the following absorptions: 8.27-8.03 (m, 3H), 4.32-4.1 (
m, 2) i).

3.72 (d, J-6Hz, 2H)*2.
22 (tt J=8 f(z.

2H), 2.27-1.68 (m, 6H), 1.42-
1.0 (m.

10H) and 0.94-0.8 (m, 6H).

Weigh the material (C, 509) on the title raw FJ of Example 8,
CH30H25rat, 20”o pa (oi-
t)2/c 90 m? (Moistened with 30% water)
The same fluffy electrostatic title product was obtained when this method was carried out using 0.249. Engineering R (nujol mul'l) 3300.
2940. 1740°1650.1540,146
8 and 1380 cm + all peaks except the last two peaks were broad and poorly resolved.

Method B The product of Example 8, Method C (30,89) was slurried in 300 mL of absolute ethanol in a 21-autoclave. 5% Pa/C1.5tl 9 (moistened with 50% water) was added and the mixture was hydrogenated at 4 atmospheres for 1 hour. During this time, hydrogen absorption was completed.

The catalyst was filtered first on paper and then on 0.45 micron nylon millibore, 100-150++ ethanol for transfer and washing! / was used. The filtrate and washings were combined and stripped to give a wet white solid. This was dissolved in 150 ml of a hot mixture of absolute ethanol and acetonitrile (1:10).

Clarified by hot filtration and boiled to 35d.
Gradual cooling to room temperature, granulation, and filtration yielded 20.19 (94%) of the crystalline, thick, non-static title product, which was then thoroughly decomposed from the main beak. Features include: 3340.3300, 2900, 2836, 1725° 1650.1628, 1580, 1532, 1455° 1410.1370, 1280, 1240, 1216a
nd 1175m Method C The crystalline product (9,41) prepared by method B above was heated to reflux for 1 hour to obtain acetone 10100
O! dissolved in it. The solution was cooled to room temperature and inoculated with a trace amount of the product of Method B to induce crystallization. After stirring for 6 hours, the purified title product was collected by filtration, washed with a minimum amount of acetone, and dried under vacuum at 35° C. (7, 259). It showed the same IR peak as the crystals from acetonitrile/ethanol in Method B.

PROCEDURE The product of Example 9 (C,509) was treated with 5% Pa10O in 125 g of absolute ethanol in a Parr hydrogenator.
, 0269C moistened with 50% water) was added. The mixture was hydrogenated under 4 atmospheres of hydrogen for 2.5 hours. Collected by catalytic filtration, Part F was subjected to IJ tubing to give the title product as a sticky solid, which was crystallized by the method of Example 9.

Example 11 N-(R-3-Methyl-4-hebutenoyl)-D The product of Example 9 (1 portion) was dissolved in 0H30f (5d).

IN NaOH (2,50 mA') is added and the mixture is stirred at ambient temperature for 3 hours. Stripped the CH30H and diluted the aqueous residue with H2O7,5at;
Extract with ethyl acetate (75agX2) and acidify with IN HCI to ptl 3.0. The acidified aqueous phase is extracted successively with fresh ethyl acetate and stripped of the extract to give the title product, which is lyophilized according to Example 10, Method A.

Example 12 Process (-trans-4-hepten-3-ol) by the method of Example 1 except that -20
The title product of Preparation Example 5 (10!;
), '0.088 mol) was converted to the title product of Example 12. After all reagents were added, the mixture was stirred at -20°C for 1.5 hours.

It was then warmed to room temperature and cooled by dropwise addition of 20 rtl of H2O and then 6 ttl of 30% NaOH (saturated with NaCe). Stir the cooled mixture for 20 minutes,
Diluted with 0820g230IILl, separated the aqueous layer and washed with fresh co2cg2 (50t/X'2). The organic layers were combined, dried over MgSOA, distilled to 50 rtl, and the residue was eluted on silica gel, first with pentane and then with pentane:
Chromatographic analysis was performed using ether (4:1).

The initial fluxin soda containing low polar impurities was discarded.

Pure product fraccine (4:1) with Rf□, 3
pentane:ether) and stripped to give the title product (5.7 g) as a clear colorless liquid.

Example 13 According to Example 3, Method A, but for isolation CH2Ce2
The product of Example 12 (5
,79,0,05m/) with ethyl orthoacetate, rearranged and saponified to give the title product (1,49) as an oil; tic Rfo, 4 (2:1
The ethyl ester of Φsan:ether) and '1 showed 1RfO,25 in the same tic system.

Example 14 The product of Example 13 (1,49) was hydrogenated over 5% Pd(OH)2/G2O,15) in 0H30H20d at 4 atmospheres for 1 hour. The catalyst was recovered by filtration.

The solvent was stripped from the P solution, leaving 8! Distillation of the product of Example 14 (1,
0! J, 0.00633-r-le) aH2cg2 1
Dissolved in oa. Oxalyl chloride (C, 547t/
, 0.00627 mol) was added and the mixture was stirred for 1 hour. During this time, gas generation ceased. The crude title product solution was evaporated under a stream of N2 to 7 d and used directly in the next step.

By the same method, the unsaturated acid of Example 13 is converted to a solution of R-3-ethyl-4-heptanoyl chloride.

Example 16 N-(S-3-ethylheptanoyl)-D-r-glutamyl (α-benzyl ester)-/IJ sil-D-alanine benzyl ester Title product of Preparation 4 (1,19,0,00222 mole)”￥ aH2ce230, d and triethylamine y (
C, 935 m/, 0.00666 mol).

C of 5-3-ethylheptanoyl chloride of Example 15
N20g2 solution (2,3a/, 0.00211 mol)
were added and the mixture was stirred for 0.75 h under N2,
0 scraps I 10% HC1, N20. Washed successively with 10% 2C03 and saturated brine, dried over MgSO4 and stripped to give a solid residue, which was triturated with ether and collected by Pweek (C.8 g). .

The product was taken up in ethyl acetate, rewashed as above, and restripped to give the secondary isomer (C, 7g), which was applied as a MV solution on silica gel to CHGI3:CH
Chromatographic purification using 30H (97:3) afforded purified product 1fi (467■).

Combining 3-ethyl-4-heptanoyl chloride 0 with the title product of Preparation 4 by the same method yields N-(R-
3-Ethyl-4-heptanoyl)-D-γ-glutamyl (α-benzyl ester)--glycyl-D-alanine benzyl ester is obtained.

Example 17 N-(S-3-methylheptanoyl)-D-γ-glutamyl-glycyl-D-alanine Title product of Example 16 (467■)y! Example 14
It was hydrogenated by the method of After recovering the catalyst, the filtrate was stripped to obtain a foam, which was placed in N20, filtered and lyophilized to yield the top product 238rn9.
I got it.

'H-nmr (DMSO-a6) delta (IIF)
: 8.20-8.00 (m, 3H), 4.24-4
．． 16 (m, 2H).

3.74-3.60 (m, 2H), z, ts (t, J=
7,2H), 2.02 (d, J=7,2H), 2.02
-1.60 (m, 3H), 1.26 (a, J=5.3H
), 1.26-1.08 (m, sH), 0.92-0.
74 (m, 5) ().

By the same method, the unsaturated product of Example 16 is converted to the same product.

Example 18 N-(R-3-ethyl-4-hebutenoyl)-D-γ-
Glutamyl-glycyl-7-ynin The unsaturated product of Example 16 was hydrolyzed by the method of Example 11,
The above title product is obtained.

Example 19 N-(S-3-ethylheptanoyl)-D-γ-glutamyl (α-benzyl ester)-glycyl - 5-3-ethylheptanoyl chloride of Example 16 and D- γ-glutamyl (α-benzyl ester)-1') sil-D-alanine butyl ester (1,0
9,0,00222 mol) to produce the title product, isolated and similarly purified. Leaf yield is 0.7
It was 9.

N-nmRDMSO-d6) delta(1)I'1
11): 8.22 (d.

J = 7t I H) + 8.12-8.00 (m
, 2H) -4,40-4,16(m, 2H), 4.
08-3.95 (m, 2H) + 3.75-3.62 (m
, 2H), 2.18 (t, J=5, 2H).

2.02 (d, J = 6, 2H), 2.04-1.62 (
m, 3H).

1.60-1.46 (m, 2H), 1.38-4.10
(m, 15H).

0.90-0.75 (m, 9H).

By the same method %R-3-ethyl-4-heptenoylchloribi
-D-γ-glutamyl (α-benzyl ester)-crysyl-D-alanine is converted to butyl ester.

Example 2O N-(S-3-ethylheptanoyl)-D-γ-glutamyl-glycyl-D-alanine butyl The title product of Example 19 was hydrogenated by the method of Example 14 to give the title product above. was manufactured.

After recovering the catalyst, strip the P liquid to obtain a solid,
The product was recovered by trituration with ether and filtration (256rIP'i).

melting point 130-131°C).

By the same method, the unsaturated product of Example 19 is converted to the same product as above.

Example 21 R-3-ethyl-4-hebutynal Example 12 in dry ethyl vinyl ether 300d
R-) lance-4-hepten-3-ol (3,09
) was heated under reflux, during which time N9(OAc) 2
5 additions <29 times, 2 hours apart). 1 more
After refluxing for 8 hours, the clear solution is cooled to room temperature, treated with 0.5 m/ml of glacial acetic acid and stirred for 3 hours. Dilute the resulting solution with ether and make 5% KOH solution 200#! / and extracted three times with ether. The ether extracts were combined, dried over K2CO3 and stripped to give the allyl vinyl ether, R-1 lance-0-vinyl-4.
-hepten-3-ol is obtained.

The allyl vinyl ether (2,3
Reflux solution 9) for 10.5 hours. The resulting solution is then cooled to room temperature and fed directly onto a silica gel column. After removing the decalin by a hexane flash, the title product is recovered by elution and stripping from the column with ether.

Example 22 The product of Example 21 (100 ml) is dissolved in 4 d of acetone and cooled to 0-5°C. In another flask, CrO3 (
72,19,0,72 mol) was mixed with 50 m/N20, stirred at 0-5°C, and concentrated H2So462.1d
The mixture was diluted with 4 N20 Y by gradually adding [.

250 d to obtain H2CrO402, 88 M-M solution (Jean's reagent) Z. This solution (1d)Y is added dropwise to the above acetone solution over a period of 1 hour. As the reagents are added, the temperature increases and is maintained at 17-25°C. 6 of the mixture
recool to 0.degree. C. and add 70 d of 2-pronozonol over 10 minutes (during which time the temperature rises to 20.degree. C.);
It is then concentrated in vacuo to give an oil, which is added with stirring to %
Add 8 ml of 2,5 N NaOH over 50 minutes and keep warm at 22±5°C. Heat the mixture, filter on diatomaceous filter, and wash with hot 2.5 N NaOH. Combine solution F and washing solution, and add isopropyl ether (10
Extract with 0mAX3). -2NN of combined organic I8
Back-extract with aOH 200πl. - In the aqueous layer (f
Acidify the aqueous layer to pH 1,0 by slowly adding lHc,e 0.5 rttl and extract the product into 300 dX3 fresh isopropyl ether. The organic extracts are combined and stripped to give the title product as an oil.

Example 23 S-7(t-butyldimethylsilyloxy)-5-methyl-1-heptene (methyl) in a 500 d four round bottom flask equipped with a stirrer, thermometer, N2 inlet and addition funnel. Triphenylphosphonium bromide (25,71J, 0.
072 mol, 1.25 equivalent t) was slurried in THF' (tetrahydrofuran) 77d and cooled in an acetone/ice water bath. n-Butyllithium (1.6 in hexane)
N solution 43.2 rtte, 0.069 m#.

1.20) Charge into the addition funnel. Initially -8℃
Addition of butyllithium to the slurry over 1 hour caused the temperature to rise and maintain the temperature at ±1°C. mixture +
Stir for an additional 0.5 h at 7Q~2°C to dissolve LiB.
Complete formation of the intermediate methylenetriphenylphosphorane in a dilute suspension of r'? I made sure. THF14m/!
The aldehyde product of Preparation Example 2 (14, IL 0.0
576 mol) was added in small portions over 40 minutes while maintaining the temperature between 3 and 7°C. After stirring for an additional 15 min, % hexane:ether (3:1)'&
No starting aldehyde was detected by the tea used (Ftf of starting aldehyde O,6; Rf of product
o, 9s).

The reaction mixture was warmed to ambient temperature and diluted with 150 ae and 90 at of ethyl acetate. The organic layer was separated and washed with H2O (100 dX2).
were combined and backwashed with 40 rtl of ethyl acetate. The organic layers were combined, dried over MgSOA, and stripped to give 25 g of an oil, which was triturated with 1 d of hexane, filtered over a sintered glass funnel, and solidified in situ. Hexane (10dX4)' was added to pulp again, and the combined hexane solution and washings were stripped to give the title product (13,59,9) as an oil.
6.6%).

1H-nmRCDC;/3) Delta (1111):
5.4-6.2 (m.

0.0 (s, 5i(CH3)2), 8 mol% (
CsHs)3Pote pollution (7,6,s, 1.25
H).

Example 24 Method A The aldehyde product of Preparation Example 6 (
26,39,0,20 mol (corrected for purity) was reacted with methylenetriphenylphosphorane. Formation of a gummy solid was observed during the addition of the aldehyde solution, but once heated to ambient temperature for processing, the reaction became a thin slurry. 82050051 reaction mixture
1 and ethyl acetate 30011A'. The layers were separated and washed with H2° (250mA!X3). - The combined aqueous layers were backwashed with ethyl acetate (300g x 2). The three organic layers were combined, dried over MgSO4, stripped and suitable for further processing in method B of fertilization.

65.7 g of an oil containing 25.69 (100%) of the title product and about 40 g of triphenylphosphine oxyto were obtained.

The pure title product is more readily obtained by conventional hydrolysis of the product of Example 23 in dilute sulfuric acid, such as in Example 26 below. The title product is isolated by extraction into ethyl acetate, drying over M9SO, and stripping.

Example 25 The title product of Example 24 (1,14 teeth, 001 mol) was dissolved in 20 g of CH220 t! Dissolved in Ll and cooled to 0°C.

Pyridine chlorochromate (4,309,0,02 mol) is added in small portions, maintaining the temperature at 0-5°C. The mixture is allowed to warm to ambient temperature, stirred for 2 hours, passed through a silica gel band, and oven stripped to give the title product as an oil, which is further purified by distillation if desired.

Example 26 Method A 2000 with stirrer, thermometer and addition funnel χ
d In a three-way round bottom flask, the title product of Example 23 (
819, corrected for purity, 0.33 mol) was dissolved in 400 alVCfn acetone and cooled to 0-5°C. In a separate flask, CrO3 (72,19°0.72 mol)
and H2O50IILeZ are mixed, stirred at 0-5[deg.]C, concentrated H2So462.1 trtl is added slowly, and the mixture is diluted with H2O to 250 g to obtain a 2.88M solution of H2CrO4 (Jean's reagent). Add 1.2 of this solution (240d, 0.67 mol) to the above acetone solution.
Added in portions over time. Upon addition of the reagents, the temperature rose rapidly to 17°C and was maintained at 17-25°C. No exotherm was observed at the end of the addition. The mixture was recooled to 6°C and 2-propatool 70d was added over 10 minutes (during which time the temperature rose to 20°C).
, then concentrated in vacuo to give an oil, to which was added 5 N NaOH with stirring and maintaining the temperature at 22±5°C.
400rrtl wokaeta. i,':+J"4tx reaction i
Fb mixture #tNr: Diluted with H2O400- and filtered through diatomaceous filter. It was pulped in moist Cake 44H2O4QOa and 50aj 5N NaOH, warmed on a steam bath and refiltered. The F solutions were combined and washed with isopropyl ether (300t/X3). - The combined M layer was back-extracted with 2N NaOH200a/. - Gradually add I#HcJ50 to the combined aqueous layer and remove it.
! (Acidified to 1,0 and the product was extracted into fresh isopropyl ether (300d x 3). The organic extracts were combined and stripped to give the title product as an oil (29.19.61 %). Secondary basic extraction of the diatomaceous earth filter cake followed by similar isolation yielded a further 7.7 g (16%) of the combined product ``H-nmRCDCe3) delta ( Lord)!
! 11.9 (s, -COOH), 5.8 (m
, -CH), 5.0(m, =CH2) and 1
．． 0 (a, -OH,)
, 3% by weight) of isopropyl ether. This contamination does not prevent the use of this product for other treatments.

Method B Example 24, Product of Method A <65.79. S-3
-Methylhept-6-en-1-ol26.39゜0
．． (containing 20 moles) tX: Oxidized according to Method A, this example. Improper tool cooled at 0-5℃ for 1
After stirring for an hour (during which time the reaction mixture turned completely green), the organic solvent was stripped, the aqueous residue was diluted with H2O250- and isopropyl ether (25
0m/X3).

When the organic extracts were combined and treated with 160-2N NaOH, a large amount of (c,H,)3po (a contaminant in the starting material) precipitated.
Washed with H. F solution 7-combined, r@7 separated, organic W
Washed with 80 m/Iy I N NaOH. Combine all aqueous layers and add isopropyl ether (250 m/×3
), acidified to pH 1, OK with concentrated HCg50-,
The desired product was purified with fresh isopropyl ether (250 dX
3) Medium pressure extraction. -Combined acidic organic extract￥M9S
Dry over O4 and strip to give the title product 14.
Obtained 0g. If desired, it is further purified by distillation.

Example 27 Pearl hydrogenation @ 10% pa/c (moistened with 50% water, 1.649), ethyl acetate 150d and unsaturated CRC of Example 26 3,289, corrected for purity) The slurry was hydrogenated under 4 atmospheres for 1.5 hours. During this time, hydrogen absorption was completed. Catalyzed by filtration at the diatomaceous top? Recover.

Furnace liquor guest stripping gave the title product as an oil (3.21.96%).

Alternatively, 5% pa/c in a 1e-autoclave
(Moistened with 50% water, 29)? The title product of Example 26 (33,
311), corrected for line spacing)! Charge, 4 atm, 3
Hydrogenation was carried out at 0 to 31°C for 2 hours, during which time hydrogen absorption was completed. The catalyst was recovered by filtration using a diatomaceous excavation, and the filtrate was collected.
Obtained, this'%: Distilled under reduced pressure to purify the title raw material (2
9,69,87,6%)? Obtained.

Boiling point: 77-79℃ 10.2nw, H-nmRC
DC/? 3) Delta (PGII): 12.0 (
s, -COO limit) 1 1.0 (a, -C character,)
.

0.6-2.8 (m, residual 13H); IR fi #
) 3400-2400, 2960, 2925, 286
0,1708.145g.

1410.1380,1295,1228,1190,
1152°1100.930m, [α:]D--6.
41' (C-1%.

) in CH30H: nl) = 1.427° Example 28 S-3-Methyl-6-heptenoyl chloride The process of Example 6 was followed by the acid product of Example 4! Title product no CH
2Cl2 solution and used directly in Example 29 below. Alternatively, the reaction mixture can be dripped,
The title product is obtained and, if desired, distilled under reduced pressure.

Example 29 N-(S-3-methyl-6-hebutenoyl)-D-γ-
Glutamyl (alpha benzyl ester')-Crystalline prepared by Method C of Example 8, FI Example 4 Product C2,779,5
mmol) was combined with the entire batch of CH2Cl2 acid chloride 9 from Example 28.

2.779 of the 1(4') initial product, recovered by washing and stripping of the dried organic layer, was taken up in 20 μl of hot ethyl acetate, this solution was diluted with 20 m6 of hexane, and the % purified product was removed by filtration. Recovered (2.2
4L77%), melting point 137.5-139.5°C.

Example 3O N-(S-3-Methyl-6-heptanoyl)-D The product of Example 29 (1 g) is eluted in CH30H5tttl. Add IN NaOH (2,50 Wtl),
The mixture is stirred at ambient temperature for 3 hours. CH30H
The aqueous residue is diluted with 7.5 ml of H2O, extracted with ethyl acetate (7,5a/, x2) and acidified to pH 3,0 with INHC,l. The acidified aqueous layer is extracted successively with fresh ethyl acetate and the extract is stripped to give the M Zhao product, which is lyophilized according to Example 10, Method A.

Example 31 N-(S-3-methyl-6-heptenoyl)-D-γ-
D-γ-glutamyl (α-benzyl ester)-glycyl-D-alanine butyl ester (Preparation Example 4) was prepared by preparing 5-3-methyl-6-naphthyl ester according to Example 8, Method A. Coupling with noyl chloride (Example 28) gives the title product above.

Example 32 N-(S-3-methylhebutenoyl)-D-7-glutamyl-glycyl-D-alaninebutylnis The product of Example 31 is converted to the title product above by the hydrogenation method of Example 10. let

Production example 1 31 equipped with stirrer, addition funnel, thermometer and nitrogen supply port! - A four-loaf flask was charged with THF400d. Cool the THF to -70°C and add ethylmagnesium bromide (214°600, d, 1.2 mol in THF).
added. While maintaining the temperature between -70℃ and -60℃,
Crotonaldehyde (78d, 0.94 mol) was processed for 7 minutes, stirred for 20 minutes at -0-70°C, then the mixture was gradually warmed to -20°C and after stirring for 1 hour, the mixture was stirred again. Cool to -70°C and saturated NH
Carefully quench with 4C7!200g (foaming at first) and warm to ambient temperature with 200% CHaCOOH and 1 % water.
Diluted with 00m/. This two-layer system is saturated with NaCβ. The aqueous layer was separated and extracted with ether (500dX2). The original stale phase and the ether extract were combined, washed with saturated NaHCO3 (250+xJX4), and the M
The residue was distilled under atmospheric pressure (699° boiling point 133-137°C).
,.

”H-nmRGDGe3) delta (pp): 5.5
3 ('+n combination m, 2H), 3.93 (m, IH), 2
．． 26(s, IH).

1.71 (d, 3H), 1.43 (m, 2l-4), 0
．． 84 (t.

3H).

Production Example 2 N-t-butyloxycarbonylglycine (109,5
1 dicyclohexylcarbodiimide (12 , 39, 60 mmol) was added and the resulting reaction mixture was allowed to warm to room temperature.

After 18 hours, the mixture was filtered and the %F solution was removed under vacuum.

The residue was dissolved in 200 d of ethyl acetate and the organic layer was diluted with 2.5 ml of ethyl acetate.
% hydrochloric acid, water, saturated hydrogen carbonate solution) and a saline solution. The organic layer was separated, dried over magnesium sulfate, and evaporated under reduced pressure. 200ml of dioxane saturated with hydrogen chloride was added to the resulting oil. After 30 minutes, 400 tttl of diethyl ether were added and the product was filtered under nitrogen atmosphere (10.99. Yield 70%
).

Production Example 3 N-t-butoxycarbonyl-D-γ-glutamyl (α
dicarbonyl-Dr-fk tamic acid α-benzyl ester (509,1431 mol) and N-hydroxy-succinamide (17,39,150 mmol) in hydrosucciniamide Add dicyclohexylcarbodiimide (30,99,15 mmol) to 1500 d of methylene chloride containing
The resulting reaction mixture was stirred at room temperature for 188 hours. The solids were filtered and the lachrymal fluid was concentrated in vacuo. The residue was triturated with diethyl ether and the solid was removed under nitrogen (43.7
9, yield 68%).

Production Example 4 Dicarbonyl-D-γ-glutamyl (α-benzyl ester) human 90-oxysuccinamide ester (4,3' If, 9.4
A solution containing glycyl-D-alanine benzyl ester salt ff (2,719,9,92 mmol) and triethylamine (1,09,9,92 mmol) was stirred at room temperature for 18 hours and then It was concentrated in vacuo. Fresh ethyl acetate 20011! The solution was washed with 2.5% hydrochloric acid, water, 10% potassium carbonate and saline solution. The organic layer was separated, dried over magnesium sulphate and evaporated under reduced pressure. The residue was treated with 200 ytl of dioxane saturated with hydrogen chloride and stirred for 2 hours. The solution was concentrated to dryness under vacuum and the residue was triturated with diethyl ether. The solid was filtered under nitrogen (3.41 overall yield 73%).

By the same method, the product of Preparation Example 3 and glycyl-D-
Alanine butyl ester is D-γ-glumemyl (α
benzyl ester)-glycyl-D-alanine butyl ester.

Production Example 5 Racemic trans-4-hepten-3-ol Production Example 1
% 2-pentenal (25 g), but using common ether as the solvent.

0.30 mol) was converted to the title product and distilled (2
5.69; boiling point 145-150°C).

Preparation Example 6 0-(t-styldimethysilyl)-8-citro stirrer, thermometer %N2 In a 500WLJ four round bottom flask equipped with a feed port and an addition funnel, S-citronellol (5471'), 3. 5 mol) of DMF (dimethylformamide)”) 547 #!/ to ambient temperature (211
? It was dissolved in :). Imidazole (262,11
1), 3.85 mol, 1.1 equivalent) was added. temperature is 1
decreased to 3°C, but further lowered to -6,5°C in an ice/acetone bath
It was lowered to . tert-butyldimethylchlorosilane (5
80.3 g, 3.85 mol, 1.1 mol) was added over 1.25 hours and the temperature was gradually increased to 11° C. over the same period. After another 0.25 hours, tgc (3
:I hexane:ether) showed complete conversion to the desired product (Rfo of starting material, 2; Rfo of product, 9
).

For isolation, mix the mixture with 500 ml of hexane and ice/
Water 1000WLI! I added it inside. The layers were separated and the organic layer was washed with ice-cold 0,25 NHC/2000 d. Combine the two aqueous layers and backwash with 500m/h of hexane.
Combine the hexane washings with the organic layer and add saturated N-He○3
500-, dried over M2SO4, and stripped to give a stoichiometric yield of the title product (986,7 g
) was obtained. This yield is theoretical due to the retention of a small amount of solvent i
It is 104% of t.

IH-nmRCD (J3) delta (F): 5.2 (
t, J=7Hz, -cH) * 3.65(t, J
-6,5Hz, -o-cdan2-)).

1.7 and 1.65 (23,2-C(Cu2)) r
O-8(st-3tC(Gdan3)3)- and 0.
0 (e, -3i (C view 3) 2) - Preparation Example 7 S-6- (t-, /Tyldimethylsilyloxy) - Mechanical stirrer, 03102 Straight glass feed tube, thermometer and etrate tube to saturation In a 500al four-hole round bottom flask with a communicating outlet, the product of Preparation Example 6 (corrected for 81.2L solvent-containing iK,
0.30 mol) of CH2G621201 and CH30H8
Dissolved in 1ml. Na HG Oa (6,
3L O, 075 mol, 0.25 equivalent Jt) was added,
The mixture was cooled to -10°C in an acetone/dry ice bath. 03102 was allowed to puzzle into the reaction mixture for 6 hours while the temperature was further lowered and maintained between -72° and 75°C. After less than L hours, no 03 was absorbed into the reaction mixture, as evidenced by the formation of a yellow color in the KI) lap. tg using hexane as acid separation liquid
c indicated that the reaction was completed (Rfo of the starting material
, 3, Rfo of the intermediate, 0) Purge excess G3 from the reaction mixture with N2 and dimethyl sulfide (26,4al, 22.49.0.3
6 mol, 1.2 eq.
no longer indicates a residual intermediate (the RtO of the intermediate
, 8: 1'(fO,05) of the product. The mixture was then stripped and the residue was washed with 150 d of ethyl acetate and 20 d of N20
300vJK was distributed. Add new H2O300d to the organic layer.
Washed with. - Add the combined N20 layer to new ethyl acetate 1
It was backwashed at 50°C. Combine organic JW1 and MgSO4
Dry on top, strip and oily! A final stripping under high vacuum for 16 hours gives a chemical yield of 74.79 ml of the title product. The yield is 101.9% of the total due to mild solvent contamination.

"H-nmRCD0g3) delta (pIP): 9.
75 (t.

-CHO), 3.6 (t, J=6H2I -0-
CH2-)I O,9(aj-8tC(Cview3)3)I
O,0(s, -51(CH3)2).

Preparation Example 8 Method A A 250 d three-necked round bottom flask was equipped with a magnetic stirrer, a thermometer, a gas supply line and a gas outlet line leading to a gas wash bottle containing saturated KI gold. 0H2C12 in this flask
S-citronellol (
31,259,0.20 mol) and cooled to -8°C. 0210 while maintaining the temperature between -2° and 10°C.
3 was allowed to puzzle into the reaction mixture for 4.5 hours. By this time, scavenging of excess 03 by the KI bath solution was seen, and mastery of the reaction was verified by a positive starch/KI-'-z''-test on the reaction mixture. was maintained at -5°C, purged with N2, and treated with methyl sulfide (
17.7tA! .

15.09. 0.24 mol) was added. The reaction mixture was warmed to ambient temperature and then stirred for 16 hours.

During this time, isopropyl ether is used as an eluent.
ea indicated that the reaction was completed (Hf of S-citronellol 0.7. Rfo of product, 4), finally the solvent was stripped to give an oil 50.89. This oil was diluted with 30 rnl of ethyl surface acid and N2025 rttl to form a single phase. Further ether 150m/
Two phases formed upon addition of .

The layers were separated and the organic layer was washed with Hzo (25υ x 2) and dried over MgSO4, stripped to a colorless oil (
23,99) was obtained. - The combined aqueous layer was diluted with ethyl acetate (5
The extracts were combined, dried and stripped to give an additional colorless oil 7.89 dX2). The colorless oils were combined and further stripped to give the title product (
28.3L (108.7% of theory) was obtained. this is(
(as mentioned above) is pure except for solvent contamination by 'cla' and is suitable for use in other reactions as mentioned above.

Method B The product of Preparation Example 2 is hydrolyzed by conventional methods, such as the dilute sulfuric acid method of Example 4 above. The title product is isolated by extraction into ethyl acetate and stripping as in the method described above.

(4 other people)

Claims (21)

[Claims] (1) Next step: (a) In the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate, the racemic trans-
4-hexen-3-ol or trans-4-hepten-3-ol is converted into unreacted trans-R-4-hexen-3-ol or trans-R-4-hepten-3- by oxidizing the S-enantiomer. (b) said trans-R-4-hexene- in the presence of acid;
3-ol or trans-R-4-hepten-3-ol is condensed with tri[(C_1-C_3)alkyl]orthoacetate to form (C_1-C_3)alkyl R-3
-Methyl-4-heptenoate or R-3-ethyl-
4-heptenoate ester; and (c) hydrolyzing said (C_1-C_3) alkyl ester to obtain said R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid. R-3-methyl-4-heptenoic acid or R-
Method for producing 3-ethyl-4-heptenoic acid. (2) Next step: (a) In the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate, the racemic trans-
4-hexen-3-ol or trans-4-hepten-3-ol is oxidized to the S-enantiomer and unreacted trans-R-4-hexen-3-ol or trans-R-4-hepten-3- (b) said trans-R-4-hexene- in the presence of an acid;
3-ol or trans-R-4-hepten-3-ol is condensed with tri[(C_1-C_3)alkyl]orthoacetate to form (C_1-C_3)alkyl R-3
-Methyl-4-heptenoate or R-3-ethyl-
obtaining 4-heptenoate; (c) ester hydrolysis of said (C_1-C_3) alkyl ester to form R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid; (d ) The 4-heptenoic acid is catalytically hydrogenated to form the S-3
- methylheptanoic acid or S-3-ethylheptanoic acid; or (e) catalytic hydrogenation of said (C_1-C_3) alkyl ester to obtain (C_1-C_3) alkyl S-3-methylheptanoate or S - producing 3-ethylheptanoate, (f) ester hydrolyzing the S-3-methylheptanoate or S-3-ethyl-heptanoate ester to produce the S-3-methylheptanoic acid or A method for producing S-3-methylheptanoic acid or S-3-ethylheptanoic acid, which comprises producing S-3-ethylheptanoic acid. (3) Claim 1 in which each of the alkyl groups (C_1 to C_3) is an ethyl group and the acid is AlCl_3.
The method described in Section 1 or Section 2. (4) Next step: (a) trans-4-hexen-3-ol or trans-4-hepten-3-ol in the presence of titanium tetraisopropoxide and L-(+)-diisopropyl tartrate; The S-enantiomer is oxidized to produce unreacted trans-R-4-hexen-3-ol or trans-R-
(b) the trans-R-4-hexen-3-ol in the presence of Hg(CH_3CO_2)_2; or by reacting trans-R-4-hepten-3-ol with ethyl vinyl ether to produce the corresponding O-vinyl ether; (c) by heating said O-vinyl ether in a solvent inert to the reaction; rearranged to R-3-methyl-
4-heptenal or R-3-ethyl-4-heptenal; and (d) producing the R-3-methyl-4-heptenal by oxidizing the 4-heptenal with chromic anhydride in a dilute inorganic acid.
A method for producing R-3-methyl-4-heptenoic acid or R-3-ethyl-4-heptenoic acid, which comprises producing 4-heptenoic acid or R-3-ethyl-4-heptenoic acid. (5) The method according to claim 4, wherein the inorganic acid is sulfuric acid. (6) Next step: (a) S-6-hydroxy-4 in a solvent inert to the reaction
-Methylhexanal or S-6-(silyl protected)hydroxy-4-methylhexanal is reacted with methylenetriphenylphosphorane to produce S-7-hydroxy-5
-Methyl-1-heptene or S-7- (silyl protected)
forming hydroxy-5-methyl-1-heptene; (b) if the product of step (a) contains a silyl protecting group, by hydrolysis of said protecting group in a dilute inorganic acid;
forming said S-7-hydroxy-5-methyl-1-heptene, this step being carried out as a separate step or carried out simultaneously with the next step; (c) forming said S-7 in a dilute inorganic acid; -Hydroxy-5-
Methyl-1-heptene is oxidized with chromic anhydride to produce S-3
- A method for producing S-3-methyl-6-heptenoic acid, comprising forming methyl-6-heptenoic acid. 7. The method of claim 6, wherein S-6-hydroxy-4-methylhexanal is the reactant of step (a) and the inorganic acid is sulfuric acid. (8) S-6-(silyl protected) hydroxy-4-hexanal is the reactant of step (a) and steps (b) and (c)
) are carried out simultaneously, the silyl protecting group is -butyldimethylsilyl and the inorganic acid is sulfuric acid. (9) Next step: (a) S-6-hydroxy-4 in a solvent inert to the reaction
-Methylhexanal or S-6-(silyl protected)hydroxy-4-methylhexanal is reacted with methylenetriphenylphosphorane to produce S-7-hydroxy-5
-Methyl-1-heptene or S-7- (silyl protected)
Forms hydroxy-5-methyl-1-heptene; (
b) if the product of step (a) contains a silyl protecting group, hydrolysis of said protecting group in dilute inorganic acid to form said S-7-hydroxy-5-methyl-1-heptene; This step is carried out as a separate step or simultaneously with the next step; (c) said S-7-hydroxy-5-
oxidation of methyl-1-heptene with chromic anhydride to form S-3-methyl-6-heptenoic acid; and (d) catalytic hydrogenation of the double bond of said 6-heptenoic acid to form S-3-methyl-6-heptenoic acid. - A method for producing S-3-methylheptanoic acid, comprising forming 3-methylheptanoic acid. 10. The method of claim 9, wherein S-6-hydroxy-4-methylhexanal is the reactant of step (a) and the inorganic acid is sulfuric acid. (11) S-6-(silyl protected)hydroxy-4-hexanal is the reactant of step (a) and step (b) and (
10. The process of claim 9, wherein c) is carried out simultaneously, the silyl protecting group is t-butylmethylsilyl and the inorganic acid is sulfuric acid. (12) The following absolute stereochemical formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, R is methyl or ethyl, and R^4 and R^
5 is each hydrogen, or one of R^4 and R^5 is hydrogen, and the other is (C_1 to C_6) alkyl or (C_6 to C_8) cycloalkylmethyl. , (a) R-trans-3-methyl-4-heptenoic acid, R
The activated form of -trans-3-ethyl-4-heptenoic acid or R-trans-3-methyl-6-heptenoic acid has the following absolute stereochemical formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R ^6 and R^7 are each benzyl, or one of R^6 and R^7 is benzyl and the other is (C_1-C_6) alkyl or (C_6-C_8) cycloalkylmethyl] (IIIc) [In the formula, one of the dashed lines (but not both) represents a double bond; When the bond is in the terminal 6,7-position, R is methyl], and (b) by hydrogenation of the compound of formula (IIIc), the double bond is removed. A method comprising the steps of simultaneous reduction and hydrogenolysis of the benzyl ester group to form a compound of formula (1). (13) The activated form of acid is acid chloride, and R^6 and R
13. The method of claim 12, wherein both ^7 are benzyl and R^4 and R^5 are both hydrogen. (14) The following absolute stereochemical formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, one of the broken lines represents a double bond, R is methyl, and R^2 and R^3 are respectively hydrogen or R^2 and R^3 each independently (C_1 to C_
6) Alkyl, (C_6-C_8) cycloalkylmethyl or benzyl; provided that the double bond is terminal 6,7-
or a pharmaceutically acceptable cationic salt thereof when R^2 and R^3 are hydrogen. (15) The compound according to claim 14, wherein R^2 and R^3 are each hydrogen. (16) The following absolute stereochemical formula: ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ A crystalline compound represented by (17) R-trans-4-hexen-3-ol or R-trans-4-hepten-3-ol. (18) The following absolute stereochemical formula: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, R is methyl or ethyl, X is hydrogen, O
R^1 is OH or Cl or forms other activated forms of the acid when X is OH and R^1 is (C_
1-C_3) alkyl]. (19) Claim 1 in which X is OH or Cl
Compound according to item 8. (20) The following absolute stereochemical formula: ▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R^9 is -CH_2OR^1^0, -CH_2
OH, -CHO, -COOH or other activated forms of COCl or the acid COOH, where R^1^0 is hydrogen or the usual silylhydroxy protecting group. (21) The compound according to claim 20, wherein R^9 is -COOH or -COCl.