PL156175B1 - A method of production of n-/s-3-alkylheptanol/-d-glutamylglycy-d-alanine and intermediate compounds used in this synthesis - Google Patents

Description

REPUBLIC

POLAND

Polish Republic

Application number: 267444

Date of notification: August 25, 1987

PATENT DESCRIPTION® PL © 156175 ® BI © I ⁿ tCl * ^:

C07K 5/04

MYTEMU

Method for the preparation of N- (S-3-alkylheptanoyl) -D- - glutamylglycylD-alanine and its esters

The holder of the patent: Priority: Pfizer Inc., New York, US August 27, 1986, US, PCT (US86) 01772 August 27, 1986, US, PCt (uS86) o1775 Proxy: PNZ "Polservice", Warsaw, PL © Application announced: July 21, 1988 BUP 15/88 The following was announced about the grant of the patent: February 28, 1992 WUP 02/92

PL 156 175 BI

1. Preparation method of N- (S-3-alkylheptanoyl) D-glutamylglycyl-D-alanine and its esters of absolute stereochemical formula 1, wherein R is a methyl or ethyl radical, both R ⁴ and R ⁵ represent hydrogen or one of them is a hydrogen atom and the second is a (C 1 -C 6) alkyl or (C 1 -C 6) cycloalkylmethyl radical, characterized in that (a) R-trans-3-methyl-4-heptenoic acid, R-trans-3-ethyl acid -4-heptenoic acid or S-3-methyl-6-heptenoic acid in an activated form is coupled with a compound of the absolute stereochemical formula 2, in which the two symbols R 6 and R ⁷ represent benzyl radicals, or one of them represents a benzyl radical and the other is a radical (C 1 -C 6) alkyl or (C 1 -C 6) cycloalkylmethyl, to give a compound of the absolute stereochemistry 3, where R, R ⁶ and R 7 are as defined above, and one dashed line, but not both, represents a double bond with the proviso that when the double bond is at the terminal 6.7 then R is ...

H 0 CH 3 H

Formula 1

C0? R ⁵

METHOD OF PREPARING N- / S-3-alkylheptanollo / -D- j-GOLlTTAMLOOlLICYL-D-AANINL AND ITS ESTERS

Claims (3)

Pateno claims 1. ^A process as preparation ^{of N} - / 3-3 alkylOH ^p-tanoilo / - ^D - ^ T-glutamyloglicylo-D-alanine and its esters of the absolute stereochemical formula steeocceernicznym 1, in which R is methyl or ety4 5 Iowa, R and R are both hydrogen atoms or one of them represents a hydrogen atom and the other represents a (C 1 -C 8) alkyl or (C 8 -C 6) cycloalkylmethyl radical, characterized in that (a) R-trans-3- Metyoo-4chppt of speech, R-tΓans-3 -ethylO4-4chepetocwy acid or s-3-eetyll-6-hepezloic acid in an activated form is coupled with a compound of the absolute steeochemical formula 2, in which both the symbols R ^D and R 'represent radicals benzyl, or one of them is a benzyl radical and the other is a (C-C) alkyl or (Cg-C /) cycloalkylnethyl radical, giving a compound of the absolute stehochemical formula 3 in which 6 7 R, R and R Raja Notices to Mariners ^e j defined and one dotted line, but not both, is a double bond, wherein when a double bond is in the end position 6, 7, then R represents a methyl radical, and / b / relationship with the above-described! The formula 3 is hydrogenated to reduce the bond with simultaneous hydrolysis of the benzyl ester groups or the benzyl ester group. 2. The method according to claim, characterized in that for the preparation _- 4 5 compound of Formula 1 in which both R and R are hydrogen atoms, activated acid, the acid chloride is reacted with a compound of formula 2, in which both the symbols R and R 'represent benzyl radicals. 3. The method according to p. A compound according to claim 1, characterized in that, in the case of the preparation of the crystalline compound of the formula 4, it is crystallized from an organic solvent or by the commission of organic solvents. The present w / rnlazku is a method wyytwwrzania immmutleh / ings N / s-3-alkiloheptazoiW 4glutamyloglicylo-D-D-3laninn and its esters of the general formula 1, wherein R OZNA ^connector ro ^{d k} n methyl or ethyl, R ¹ and ^r1 are both hydrogens, or it en ^d of symtaoM R ¹ and ^{R 1} is hydrogen mark and dru ^g and ^k represents degeneration ^{/ C} i-Cg ^g -alk ^of head lu ^b / ^ -C ^ CC cyktaolkHometylowy. The relatively new field of immeutOartmriOol / i., Especially the department dealing with immunomooulation, is developing rapidly. Numerous compounds of natural origin have been investigated, among them the tetrapept, called tuftsynr ^, i.e. n / 2 - L-threonyl-Lolysyl Z-3 prolyl 7-L · -rgnite. Much attention has been given to the synthesis of peptidoglycan derivatives, especially to compounds known as mLu-amyl dipeptides. The subject of the invention is a method of annealing N- (S-3-allylheptatooyl) -D-glutamylglycyl-D-αlatine and its steeoicheπ] ic esters 1, where R is a methyl or ethyl radical, both symbols R and R represent hydrogen atoms, or one of them represents a odor atom, and the other is a / C 8 -alkyl or / C 8 -C 8 / cycloalkyl-methyl radical, which consists in the fact that / a) R-tans-3-methyl- 4-hephtoic acid, R-tans-3-ethylO444chepttlCwy or S-3-methyl-6-hepetonic acid in an activated form is coupled with a compound with an absolute value and a chemical concentration of 2, in which both the symbols R ° and R ¹ represent radicals benzyl groups, or one of them is a benzyl radical and the other is a (C 1 -C 8) alkyl radical or (C 8 -C 6) cyylalkyl reliethyl radical, which gives a compound of the absolute formula stehlichemical. e 3, in which 156 175 R ^k and R ⁷ paradise ^± defined above, and one of the dotted line, but not both, is a double bond, wherein when a double bond is in the end position 6, 7, then R represents a methyl radical, and / b / the compound of above described in C 1-6 is hydrogenated by reducing the double bond with simultaneous hydrolysis of the benzyl ester groups or the benzyl ester group. Preferably, an acid chloride is used as the activated form of the acid and a compound of formula II in which R and R are benzyl radicals is used as the compound of formula II. The result is Ζ 5 of a compound of formula 1 in which R and R are hydrogen. In one particular case, the process according to the invention results in a crystalline compound having the absolute steroochemical formula 4, which consists in carrying out the crystallization of the compound obtained according to the invention from an organic solvent or from a commission of organic solvents. According to the method of the invention, unsaturated acids in active form, e.g. as an acid chloride, either as a conventional anhydride mixture, or activated with a conventional dehydrating coupling agent such as dίoylohrxylcarbodiieid, are coupled in the usual manner with a compound of wt i-ze 2 to obtain a compound of formula 3. The compound of formula III is then hydrogenated to give an immaterial compound of formula 1. During this hydrogenation, the double bond is saturated and the benzyl group or groups are hydrolyzed. The hydrogensinir is reacted in an inert solvent in the presence of a catalyst, e.g. nickel or a supported noble metal such as Ranney nickel or Pd / c, or without a support, e.g. RWH3. The type of solvent and temperature are not critical. Suitable solvents are lower alcohols, ethers, e.g. dioxane, tetrahydrofuran and dimethyethases, and esters, e.g. ethyl acetate. Preferably room temperature is used, but in the case of an exothermic reaction, cooling may not be used so as not to increase the cost of the process. The pressure is also not of decisive importance and, in order to avoid the expensive equipment of the WLe, a pressure of less than 7000 hPa is used. Hydrogenation with palladium on carbon is preferably carried out at a pressure of 3-6 times the atmospheric pressure. , 4 5 The compound of formula I, in which R is methyl and R and R are hydrogen, are preferably obtained in crystalline form by crystallization from an organic solvent or a mixture of such solvents. Suitable solvents are acetone, acetonitrile with ethanol, and tetrahydrofuran with ether. In view of the efficiency of the crystallization process, a mixture of acetonitrile and ethanol is the preferred solvent, but acetone is more preferable for the purity of the product. This new form of this compound is much more valuable than the known product in the form of liofll · izivanegd. The product in this new form is more compact, easy to minimize and less electrostatic, so that it can be used in various ways. The following examples illustrate the method in the woług whmlazku. EXAMPLE 1 N- (S-3-meyyl-4-heptenoyl) -D-1-glutamid (c. Bzyloxy) carbonate benzyl ester ^of d (glycidyl-D-alaniase). In a 500 ml four-neck round bottom flask equipped with a stirrer, thermomeer, incubator and nitrogen inlet, 32.8 g (0.059 moles) of the compound obtained in the manner described in Example 9 are dissolved in 175 ml of CH ^ 1 g, cooled to the temperature of ^y 0- ⁵ ° ^C and maintained c ^e tempeΓatuΓę ^adding stΓ ^ free ^ ^Mirza, for ¹ 5 minutes, 24.7 ml, i.e. 17.9 g / 0.177 mooa 3 równowianZki / tΓÓjenydaaπlizn. Keeping the mixture still in the ice-water bath, all of the solution of S1-methylheptenoyl chloride in CHgCl3, same as in Example 10, was added over 15 minutes. The temperature of the mixture was raised to 21 ° C and the mixture was rubbed without removing the cooling bath. continue for 45 minutes until dndla, when the mixture becomes gelatinous and does not mix. The obtained race is ground, mixed with 125 ml of 10% HCl and 50 ml of CHgClg, the organic layer is separated, washed successively with 2 x 125 ml of water, 2 x 125 ml of 10% K ^^ O, and 125 ml Water, then dried over MgSO 4 and the solvent removed to give a white solid. This product was dissolved in 500 ml of hot ethyl acetate and the solvent was stripped, causing the title product to crystallize. Example II. : \ '- (S-3-netylheptar.oyl) -D- < E > glycuaarnylglycyl-D-alanine. 30.8 g of the product obtained according to Example 1 are mixed in 300 ml of anhydrous ethanol in a 2-liter autoclave, then 1.54 g of 5% Pd / θ with a water content of 505 are added and the mixture is hydrogenated at 4050 hPa for 1 hours until the uptake of hydrogen is complete. The catalyst is then filtered off, first through filter paper and then through 0.45 micron nylon, using 100-150 ml of ethanol to filter and rinse. The filtrate with the stream is evaporated and the wet white solid residue is dissolved in 150 ml of a hot mixture of anhydrous ethanol with acetonitrile 1:10, then filtered hot, evaporated to a volume of 35 ml, cooled to room temperature, granulated and filtered to give 20. 1 g / 94% of theory / a crystalline, compact and non-electrosaatic product. This product has an ir spectrum (powder in nujol) with well defined sharp peaks at 3340, 3300, 2900, 2836, 1725, 1650, 1629 ^, 1580 ^, 1532 ^, 1455 ^, 1410, 1370 ^, 1280 ^, 1240, 1216 and 1175 ^{cm -1} . 9.4 g of the crystalline product obtained as described above are dissolved in 1000 ml of acetone and refluxed for 1 hour, then the solution is cooled to room temperature and seeded with a trace of crystals of the product in order to cause crystallization. . After stirring for 6 hours, the precipitate is filtered off, washed with a small amount of acetone, and dried in vacuo at 35 ° C to give 7.25 g of product, which in the ir spectrum shows peaks identical to the product obtained from acetonitrile / ethanol as described above. . Example III. N- (S-3-ethyl-4-heptenoyl) -D-ff-glutornyl / cC be.ocyloxycarbonyl o / -glycyl-D-rlarine benzyl ester. 1.1 g (0.00222 mole) of the product obtained in Example 9 are dissolved in 30 ml of CH 2 Cl 2, 0.935 ml (0.00666 mol) of thymethylamine is added, followed by the addition of 2.3 ml (0.00211 mol) of chloride. S6-ethylheptenoyl and stirred under a nitrogen atmosphere for 0.75 hours. Then the mixture is washed successively with 20 ml of 10% HCl, water and 10% K 2 CO 3 solution and brine, dried over MgSO 4 and evaporated. The solid residue is triturated with ether and filtered off. Example IV. N- (S-3-ethyl-4-heptenoyl) -D-yL-glutameloglycyl-D-alanine. 467 mg of the title product of the previous example are hydrogenated by the method described in Example 2 and, after the catalyst has been filtered off, the filtrate is evaporated. The foamy residue is dissolved in water, filtered and lyophilized to give the title product. ^Example V. ^E ster tonzylowy N - ^ - ^ me-methyl-c-lieptenoilo / - ^- - glutamyl-iT / benzyloksylkιrbonnlo / -glycyl-D-rlaniny. Using the method of Example 1, 2.77 g (5 mmmH) of the product of Example 9 was coupled with the acid chloride in CH 2 Cl 2. The filtered organic solution was washed with water, dried and evaporated to give 2.77 g of the product, which was dissolved in 20 ml. hot ethyl acetate, dilute the solution with 20 ml of hexane, cool and filter the precipitate to give 2.24 g (77% of theory) of the title product, melting at 137.5-139.5 ° C. . Example VI. N- (S-3-eeiylheptanoyl) -D-glutamylglycyl-D · -alanine. In a similar manner to that described in Example 2, the product of the previous example is converted into the title compound. Example VII. Glycyl-D-alanine benzyl ester hydrochloride. For cold / 0 ° C / solution 10 g / 57 mimoi / ^ ^τ -III-Γboot Ooksykrrlti> nylogicin, 20 g / 57 mimoi / p-toluenesilphonate of benzyl ester D - alanine and 5.77 g / 57 mrally / triets of Oαamines in 100 ml of methylene chloride, 12.3 g (60 mmole) of awncyclohexylcarbodiimide are added and the mixture is allowed to warm to room temperature. After 18 hours, the mixture is filtered, the filtrate is evaporated under reduced ηΐβηίθπι, the residue is washed with 2.5% Hydrochloric acid, water, saturated sodium bicarbonate solution and brine, then the organic solution is dried over MgSO 4 and evaporated under reduced pressure. 200 ml of dioxane saturated with hydrogen chloride are added to the oily residue, and after 30 minutes, 400 ml of diethyl ether are added. After filtering off the precipitate under nitrogen atmosphere, 10.9 g of product of theory are obtained. Example VIII. Hydroxysuccinic acid amide N-tertiary ester of uuook S-carbonyl-D-f-glutamyl (oC-benzyl). To 1500 ml of methylene chloride containing 50 g / 143 mmooe / N-tertiary acid / cC- benzyl ester / Uuloxyc ^ Γbonslσ-D- ^ Γ-glutamate and 17.3 g / 150 mimol / N-hydroxysuccinimide are added 30.9 g (15.5 H) of dicyclohexylcarbodiimide were added and the resulting mixture was stirred at room temperature for 18 hours. The precipitate is then filtered off, the filtrate is evaporated under reduced pressure and the residue is triturated with diethyl ether. After filtering off the precipitate under nitrogen, 43.7 g (68% yield) are obtained. When ^k order IX. Hydrogen chloride acid ^{k b} enz ^y lowe ^g of D- ^ -glutamylo- / C ^C- bbenzyloks carbonyl ^y / -glycyl-D-alanine. A solution of 4.3 g (9.45%) of N-tertiary buoxycabbonyl-D- Jf-glutamyl / σ6bieric acid / hsdroxsbursyn amide, 2.71 g (9.92 nevi) benzyl glycyl-D- ester hydrochloride alanine and 1.0 g (9.92 imoyl) triethylumine in 100 ml of methylene chloride were stirred at room temperature for 18 hours and then evaporated in vacuo. The residue is dissolved in 200 ml of ethyl acetate, the solution is washed with 2.5% hydrochloric acid, 10% potassium carbonate solution and brine, dried over NgSO4 and evaporated under reduced pressure. The solid is mixed with 200 ml of dioxane saturated with hydrogen chloride, stirred for 2 hours and evaporated to dryness in vacuo. The residue is triturated with diethyl ether and filtered off under nitrogen to give 3.41 g (73% yield) of the product. In an analogous manner, the product of the preceding example and the glycyl-D-alanine butyl ester are converted to D-1 -L-glutamyl / C-benrsyloxy-carbonyl-D-glycyl-D-alanins ester. Example X. S-3-Methylheptenoyl chloride. 8.5 g (0.062 moles) of S-3-methyl-4-hepenoic acid are dissolved in 18 ml of CHgCl, and 5.36 ml (7.80 g, 0.0614 moles) of oxalyl chloride are added and the imesanine is left for 4 hours, until the reaction is complete, as evidenced by the cessation of gas leakage. The obtained acid chloride solution is used directly in Example 1. Example XI. S 1 -methyl 4 -heptenoyl chloride. 0.747 g (5 mml) of S-3-methyl66-hepeenoic acid is converted to a solution of the title compound at C5C2 using the method set out in Example 10 and used directly in Example 5.