KR840000010B1 - Method of preparing for muranyldipeptide derivatives - Google Patents

Abstract

Muramyldi-peptide derivatives (I) are prepared from compound (III) reacted with active ester of aliphatic acid, or from compound (II) reacted with alkyl-diamine (H2N-C(R1)H-(CH2)n-NHCO-A) of alkylamine (H2N-A) or amino group acyllized; where x= L-alanine, L-cerine, L-barine or glycine; Y= -NH-A or -NHC(R1)H(CH2)n-NHCO-A (where R1 = h atom, C1-C6 alkyl, carboxamide or carboxyl group, n = 1-6, and A = C7-C30 chained aliphatic hydrocarbon); and acy1 = acy1 group of C2-C6 aliphatic carboxylic acides.

Description

Method for Preparing Muramil Dipeptide Derivatives

The present invention relates to a method for producing a muramyl peptide derivative of the following structural formula (1) effective for the prevention and treatment of infectious bacteria.

In the above structural formula

X is an amino acid residue (e.g., L-alanine, L-serine, L-valine, glycine, etc.)

(여기에서 R₁은 수소원자, 탄소수 1 내지 6의 알킬그룹, 카복스아미드 그룹 또는 카복실그룹이고 n은 1 내지 6의 정수이며Y is -NH-A or

Wherein R ₁ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a carboxamide group or a carboxyl group and n is an integer of 1 to 6

A is a straight or branched, saturated or unsaturated aliphatic hydrocarbon residue having 7 to 30 carbon atoms and "acyl" is an acyl group of aliphatic carboxylic acid having 2 to 6 carbon atoms.

A characteristic feature of the compounds according to the invention is the peptide bond with the terminal amino group (isoglutamine) with an alkylamino group (NH-A) or a dialkylamino group in which one of the amino groups is acylated.

In recent years, we have observed the progress and prognosis of patients with leukemia, malignant lymphoma, various carcinomas or metabolic deficiencies in organs. It has been found that they tend to occur frequently.

Despite the improved selection of bacteria involved in treatment, improved preventive measures against infection and the widespread use of several chemotherapeutic agents, the results from common infections and incurable diseases were death. This situation led to damage of the resistant factor against infection by phagocytes such as polymorphonuclear leukocytes. Damage to the resistant factor not only reduces the host's defense against the infecting bacterium itself, but also the defense of iatogenic agents such as anticancer agents, immunosuppressants and paracortex. Therefore, the exploration of drug development having a prophylactic and therapeutic effect against the above-mentioned infectious bacteria has increased. However, in the present situation, the problem is caused by the resistant bacteria obtained by frequently using the chemotherapeutic agent treated with the infectious bacteria using a weakly toxic bacterium developed by the bacterial change phenomenon, so it has no direct effect on the bacterial resistance. The development of new types of drugs with prophylactic and therapeutic effects has emerged.

Some examples known to exhibit these effects and actions include bacterial whole cells, cytoskeleton and extracts such as lipopolysaccharides or cord factors, such as mycobacteria, corynebacteria, streptococcus and listeria. Derived from microorganisms. However, these bacteria cannot be used in the human body. This is because of side effects such as immunity and purulent.

It has been reported that N-acetylmuramil-L-alanyl-D-isoglutamine (hereinafter referred to as muramildipeptite) has a protective effect against certain infectious organisms (see Proc. Natl. Acad. Sci. USA Vol. 74, 5, pp. 2089-2093). However, the effects exhibited in these compounds are not satisfactory enough. PCT WO 79/00802 also describes muramyldipeptide derivatives, but these compounds differ in structure from the compounds of the present invention.

The effect of the compound of the present invention was confirmed by the method described below.

1. Preventive effect against microbial infection In 20 rats of each group (weight 26g ± 1g, 5 weeks old) subcutaneously injected 0.1ml each of compounds 1 to 7 and control compounds 1 to 4 of the present invention E after 24 hours Coli E77156 strains were inoculated subcutaneously at doses of 6 × 10 ⁶ , 9 × 10 ⁶ and 1.2 × 10 ⁷ , respectively. Efficacy is assessed as survival rate of the juju after 7 days of infection. It is evident from the results of Table 1 that the compound of the present invention has an excellent prophylactic effect against infecting bacteria.

TABLE 1

Preventive effect on strains infected with E. Coli E 77156

* Survival shows the difference between the groups treated with the compounds of Table 1 and the untreated groups.

2. Auxiliary action

According to the method by Coinventer, Kotani et al. (Biken Journal, Vol. 20, 95 103, 1977), a Conyl test was conducted to measure the co-operation index using a mole mote. The results are shown in Table 2 and showed that the compounds of the present invention are superior to the co-action of muramdipeptides.

TABLE 2

Conil test

Control compounds 1 to 4 and compounds 1 to 11 of the present invention described in Tables 1 and 2 are as follows.

(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-L-라신,Control Compound 1:

(N-acetylmuramil-L-alanyl-D-isoglutaminyl) -L-lacin,

Control compound 2: N-acetylmurayl-L-alanyl-D-isoglutamine,

Control compound 3: N-acetylmurayl-L-seryl-D-isoglutamine,

Control compound 4: N-acetylmurayl-L-valyl-D-isoglutamine,

-(N-아세틸무라밀-L-알라닐-D-이소 글루타미닐)-

-옥타 노일-L-리신,Inventive Compound 1:

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Octanoyl-L-lysine,

-(N-아세틸무라밀-L-알라닐-D-이소 글루타미닐)-

-라우 로일-L-리신,Inventive Compound 2:

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Lau Lauyl-L-Lysine,

-(N-아세틸무라밀-L-알라닐-D-이소 글루타미닐)-

-스테아 로일-L-리신,Inventive Compound 3:

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Stearoyl-L-lysine,

-(N-아세틸무라밀-L-세릴-D-이소 글루타미닐)-

-스테아 로일-L-리신,Inventive Compound 4:

-(N-acetylmurayl-L-seryl-D-isoglutaminyl)-

Stearoyl-L-lysine,

-(N-아세틸무라밀-L-발릴-D-이소 글루타미닐)-

-스테아 로일-L-리신,Inventive Compound 5:

-(N-acetylmurayl-L-valyl-D-isoglutaminyl)-

Stearoyl-L-lysine,

-(N-아세틸무라밀-L-알라닐-D-이소 글루타미닐)-

-트리아콘타 노일-L-리신,Inventive Compound 6:

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

-Triconta noil-L-lysine,

-(N-아세틸무라밀-L-알라닐-D-이소 글루타미닐)-

-이소펜타 데카노일-L-리신,Inventive Compound 7:

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Isopenta decanoyl-L-lysine,

Inventive Compound 8: N-acetylmurayl-L-alanyl-D-isoglutamine octylamide,

Inventive Compound 9: N-acetylmurayl-L-alanyl-D-isoglutamine laurylamide,

Inventive Compound 10: N-acetylmurayl-L-alanyl-D-isoglutamine stearylamide and

Inventive Compound 11: N-acetylmurayl-L-valyl-D-isoglutamine stearylamide.

The compound of the present invention can be prepared according to the following scheme.

In the above structural formula

n, acyl, R ₁ , A, X and Y are as defined above.

In particular, the compound of formula (I) of the present invention is a method of reacting muramic acid dipeptide (II) with a compound of formula (IV) (method a) or reacting tripeptides (III) of muramic acid with a compound of formula (V) It can manufacture by (b).

(I)의 반응은 펩타이드 합성에서 일반적으로 사용되는 방법 (예, 카보디이미드법, 활성 에스테르법 산무수물법등)에서 선택할 수 있다. 예를 들면 축합 반응은 구조식(Ⅱ) 및 (Ⅳ)화합물을 N,N-디메틸-포름아미드(이하 DMF라 칭함), 테트라하이드로푸란 또는 이의 혼합물중에 용해 또는 현탁시킨다음 N-하이드록시석신이미드, 1-하이드록시벤조트리아졸, N-하이드록시-5-노르보르넨-2,3-디카복스이미드 또는 펜타클로로페놀등에서 선택한 시약(i) 및 디사이클로 헥실 카보디이미드, 1-에틸-3-(3-디메틸아미노프로필) 카보디이미드 또는 이의 염산염과 같은 카보디이미드(ii) 존재하에 실온 내지 60℃의 온도에서 약 3시간 내지 2일간 반응시킴으로써 수행될 수 있다.When using method a condensation reaction, ie (II) + (IV)

The reaction of (I) can be selected from methods generally used in peptide synthesis (eg, carbodiimide method, active ester method, acid anhydride method, etc.). For example, the condensation reaction dissolves or suspends the compounds of formulas (II) and (IV) in N, N-dimethyl-formamide (hereinafter referred to as DMF), tetrahydrofuran or mixtures thereof and then N-hydroxysuccinimide , Reagent (i) selected from 1-hydroxybenzotriazole, N-hydroxy-5-norbornene-2,3-dicarboximide or pentachlorophenol and dicyclohexyl carbodiimide, 1-ethyl-3 In the presence of carbodiimide (ii) such as-(3-dimethylaminopropyl) carbodiimide or hydrochloride thereof.

When the reaction of (III) + (V)-(I) is used in the condensation reaction of method (b), the reaction conditions used in method (a) can be similarly used. For example, the compound of formula (V) may be dissolved or suspended in tetrahydrofuran, chloroform or a mixture thereof and then N-hydroxysuccinimide, 1-hydroxybenzotriazole, N-hydroxy-5norbornene-2 In the presence of carbodiimide with a reagent selected from, 3-dicarboximide, pentachlorophenol and the like at room temperature to 60 ° C. for several hours or overnight, an active ester of formula (V) is obtained. It can be condensed.

The condensation reaction is generally carried out with stirring for several hours or two days at 0 to 60 ° C., preferably at 25 to 40 ° C., as a solvent or a mixture used in peptide synthesis, for example, DMF, tetrahydrofuran, water, chloroform, or the like. can do.

The invention is illustrated by the following examples and preparation methods.

Example 1

-(1-α-0-벤질-4,6-0-벤질리덴-N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-벤질옥시카보닐-L-리신 벤질에스테르를 115ml의 60% 아세트산에 현탁시키고 끓는 수욕상에서 약 1시간동안 가열한다. 실온으로 냉각한 후 반응혼합물을 감압하에 농축시키고 잔사에 물을 가한다. 침전된 결정을 여과하여 모으고 DMF-에틸아세테이트로 재결정시켜 7.6g의

-(1-α-D-벤질-N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-벤질옥시카보닐-L-리신 벤질 에스테르를 얻는다. 융점 : 207 내지 210℃ (분해).11.4 g

-(1-α-0-benzyl-4,6-0-benzylidene-N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Benzyloxycarbonyl-L-lysine benzyl ester is suspended in 115 ml of 60% acetic acid and heated for about 1 hour in a boiling water bath. After cooling to room temperature, the reaction mixture is concentrated under reduced pressure and water is added to the residue. The precipitated crystals were collected by filtration and recrystallized with DMF-ethyl acetate.

-(1-α-D-benzyl-N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Obtain benzyloxycarbonyl-L-lysine benzyl ester. Melting point: 207-210 degreeC (decomposition).

＋56.6°(C=0.3, DMF, 17시간후).

+ 56.6 ° (C = 0.3, DMF, after 17 hours).

H₂OElemental analysis: C ₄₇ H ₆₂ O ₁₄ N ₆

H ₂ O

Calculated Value (%): C 59.80, H 6.73, N 8.90

Found (%): C 59.65, H 6.67, N 8.96

The compound obtained above is dissolved in 20 times acetic acid and hydrogenated at room temperature in the presence of palladium-carbon under hydrogen stream, then the catalyst is filtered off and the filtrate is concentrated under reduced pressure. The residue is purified by chromatography on silica gel using n-butanol-acetic acid-water (4: 1: 1 in volume ratio) as eluent. The fraction containing the desired compound is concentrated under reduced pressure and treated with a basic ion exchange resin (acetate type). The eluate is lyophilized to yield N ^α- (N-acetylmuramil-L-alanyl-D-isoglutaminyl) -L-lysine.

-(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-옥타노일-L-리신을 수득한다.300 mg of N ^α- (N-acetylmuramil-L-alanyl-D-isoglutaminyl) -L-lysine in 7 ml of DMF and ice-cooled with 148 mg of octanoic acid active N-hydride Roxy-5-norbornene-2,3-dicarboxamide ester and 0.05 ml of N-methylmorpholine are added. After 30 minutes, the mixture is gradually warmed to room temperature and stirred overnight. The reaction mixture is concentrated under reduced pressure and diethyl ether is added to the residue. The precipitated crystals are collected by filtration and washed successively with diethyl ether and water. 310 mg of recrystallized from DMF-diethyl ether

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

-Octanoyl-L-lysine is obtained.

Melting Point: 126 ~ 130 ℃ (Decomposition)

＋28.8°(C=0.2, DMF 17시간후).

+ 28.8 ° (C = 0.2, after DMF 17 hours).

Elemental analysis: C ₃₃ H ₅₈ O ₁₃ N ₆ 2H ₂ O

Calculated Value (%): C 50.63, H 7.98, N 10.74

Found (%): C 50.32, H 7.54, N 11.00

The following compounds are prepared in the same manner as above.

-(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-라우로일-L-리신,

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Lauroyl-L-lysine,

Melting Point: 174.5 to 176.5 ° C. (Decomposition)

＋30.2°(C=0.6 DMF, 17시간후).

+ 30.2 ° (C = 0.6 DMF, after 17 hours).

Elemental _{analysis: C 37 H 66 O 13 N} 6 · H 2 O

Calculated Value (%): C 54.13, H 8.35, N 10.24

Found (%): C 53.90, H 8.40, N 10.17

-(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-스테아로일-L-리신

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Stearoyl-L-Lysine

Melting point: 175.0 to 177.0 ° C. (decomposition).

＋25.9°(C=0.6, DMF, 17시간후).

+ 25.9 ° (C = 0.6, DMF, after 17 hours).

Elemental _{analysis: C 43 H 78 O 13 N} 6 · H 2 O

Calculated Value (%): C 57.06, H 8.91, N 9.29

Found (%): C 57.21, H 8.87, N 9.19

-(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-트리아콘타노일-L-리신

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

-Tricontanoyl-L-lysine

Melting point: 165.0-168 ° C. (decomposition).

H₂OElemental analysis: C ₅₅ H ₁₀₂ O ₁₃ N ₆

H ₂ O

Calculated Value (%): C 61.03, H 9.78, N 7.76

Found (%): C 60.89, H 9.58, N 7.63

-(N-아세틸무라밀-L-알라닐-D-이소글루타미닐)-

-이소펜타데카노일-L-리신

-(N-acetylmurayl-L-alanyl-D-isoglutaminyl)-

Isopentadecanoyl-L-lysine

Melting Point: 144.5 ~ 147.5 ℃ (Decomposition)

＋20.2°(C=0.3, DMF, 17시간후).

+ 20.2 ° (C = 0.3, DMF, after 17 hours).

Elemental analysis: C ₄₀ H ₇₂ O ₁₃ N ₆

Calculated Value (%): C 56.85, H 8.59, N 9.95

Found (%): C 57.26, H 8.63, N 9.81

-스테아로일-L-리신N ^α- (N-acetylmurayl-L-seryl-D-isoglutaminyl)-

Stearoyl-L-Lysine

Melting Point: 175.0 to 177.0 ° C (Decomposition)

＋27.8°(C=0.5, DMF, 17시간후)

+ 27.8 ° (C = 0.5, DMF, after 17 hours)

H₂OElemental analysis: C ₄₃ H ₇₈ O ₁₄ N ₆

H ₂ O

Calculated Value (%): C 55.52, H 8.78, N 9.04

Found (%): C 55.46, H 8.62, N 9.06

-(N-아세틸무라밀-L-발릴-D-이소글루타미닐)-

-스테아로일-L-리신

-(N-acetylmuramil-L-valyl-D-isoglutaminyl)-

Stearoyl-L-Lysine

Melting Point: 182.0 ~ 185.0 ℃ (Decomposition)

＋22.8°(C=0.5, DMF, 17시간후).

+ 22.8 ° (C = 0.5, DMF, after 17 hours).

H₂OElemental analysis: C ₄₅ H ₈₂ O ₁₃ N ₆

H ₂ O

Calculated Value (%): C 58.48, H 9.05, N 9.09

Found (%): C 58.48, H 8.89, N 9.12

Example 2

300 mg of N-acetylmural-L-alanyl-D-isoglutamine was dissolved in 2 ml of DMF, and 72 mg of octylamine, 64 mg (0.56 mmol) of N-hydroxysuccinimide and 7 ml of tetrahydrofuran Add. 115 mg (0.56 mmol) of dicyclohexylcarbodiimide are added thereto while cooling with ice and stirring. After 30 minutes the mixture is gradually warmed to room temperature and allowed to react overnight. The precipitated dicyclohexylurea is filtered off, the filtrate is concentrated under reduced pressure and water is added to the residue to give a precipitate. Recrystallization of the precipitate with DMF-diethyl ether gave 165 mg of N-acetylmurayl-L-alanyl-D-isoglutamine octylamide.

Melting Point: 174.0 to 175.5 ° C. (Decomposition)

＋30.5°(C=0.4, 아세트산, 21시간후).

+ 30.5 ° (C = 0.4, acetic acid, after 21 hours).

Elemental _{analysis: C 27 H 49 O 10 N} 5 · H 2 O

Calculated Value (%): C 52.15, H 8.26, N 11.26

Found: (%): C 51.88, H 8.00, N 11.35

Preparation of active esters of fatty acids.

1.0 mmol of fatty acid is dissolved in 10 ml of tetrahydrofuran, cooled with ice and stirred with 206 mg of dicyclohexylcarbodiimide and 179 mg of N-hydroxy-5-norbornene-2,3-dicarboximide Add. After 30 minutes the mixture is gradually warmed to room temperature and allowed to react for 5 hours. The precipitated dicyclohexyl urea is filtered off and the filtrate is concentrated. Anhydrous diethyl ether is added to the residue, and the insolubles are filtered off. The filtrate is concentrated under reduced pressure and dried to give the active N-hydroxy-5-norbornene-2,3-dicarboximide ester of fatty acid as white crystals.

When preparing an active ester of triacontanoic acid, a mixed solvent of tetrahydrofuran and chloroform (1: 1 in volume ratio) is used as a reaction solvent.

Although the present invention has been described in detail in the specification, those skilled in the art may make changes and modifications within the scope of the present invention.

Claims (1)

Reacting a compound of formula (III) with an active ester of a fatty acid (A-COOH) or a compound of formula (II) acylated with an alkylamine (H ₂ NA) or an amino group

Reacting with a method for producing a muramyldipeptide derivative of formula (I).

In the above structural formula X is the amino acid residue of L-alanine, L-serine, L-varin or glycine Y is -NH-A or

Wherein R ₁ is a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a carboxamide group or a carboxyl group, n is an integer of 1 to 6 and A is a straight or branched, saturated or unsaturated aliphatic hydrocarbon having 7 to 30 carbon atoms "Acyl" is an acyl group of aliphatic carboxylic acid having 2 to 6 carbon atoms.