KR810001525B1 - Process for the preparation of peptide derivatives - Google Patents

Abstract

Peptide derivs.(I; R1 = H, methyl, hydroxymethyl, mono-, di-or trihalomethyl; R2 = lower alkyl, hydroxy-(lower alkyl) or guanidino(lower alkyl), useful as bactericides, were prepared by the reaction of II(R10 = R1 or protected hydroxymethyl; R3, R4 = H or lower alkyl protecting group) and III(R5 = protected amino group; R20 = R2 or protected hydroxy-(lower alkyl) or guanidino-(lower alkyl) group.

Description

Method of Preparation Peptide Derivatives

The present invention relates to a method for preparing a peptide derivative of the following formula (I) useful as an antibacterial agent.

In the above structural formula

R¹ is hydrogen, methyl or hydroxymethyl or mono-, di- or trihalomethyl

R² is lower alkyl, hydroxy- (lower alkyl) or guanidino- (lower alkyl) that is not a characteristic group of a-amino acids in the form normally found in proteins

The arrangement at the carbon atom represented by (a) is (R) when R¹ is not hydrogen and

In the carbon atom represented by (b), the arrangement is (L).

"Lower alkyl" is straight-chain or branched alkyl having up to 8 carbon atoms, and includes ethyl, propyl, butyl, tertiary butyl, pentyl, hexyl and the like.

Examples of hydroxy- (lower alkyl) represented by R² are 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxybutyl and the like.

“Halo” is fluoro, chloro, bromo, urethra, and examples of halomethyl are chloromethyl, dichloromethyl, trifluoromethyl and the like.

In the formula (I), when R¹ is not hydrogen, the arrangement at the carbon atom represented by (a) is (R). Namely, it is an arrangement obtained by substituting a carboxyl group of natural α-amino acid with a phosphorus site.

Preferred peptide derivatives prepared by the present invention are compounds of formula (I) wherein R 1 is hydrogen or methyl and pharmaceutically nontoxic salts thereof. More preferred are compounds of formula (I) wherein R2 is lower alkyl, in particular ethyl, n-propyl or n-butyl.

The compound of the above-mentioned structural formula (I) is as follows.

(1R) -1- [L- (α-aminobutyryl) amino] -ethylphosphonic acid,

(1R) -1- (L-norroylamino) -ethylphosphonic acid,

(1R) -1- (L-norvalylamino) -ethylphosphonic acid,

(1R) -1- [L- (α-aminobutyryl) amino] -methylphosphonic acid,

(1R) -1- [L-norroylamino] -ethylphosphonic acid,

(1R) -1- (L-norvalylamino) -methylphosphonic acid,

(1R) -1- (L-homoarginylamino) -ethylphosphonic acid and

(1R) -1- (L-homoarginylamino) -ethylphosphonic acid.

According to the present invention, the peptide derivative of formula (I) and pharmaceutically non-toxic salt thereof are present in a protecting group or condensation product after condensation of a compound of formula (II) with a protected α-amino acid of formula (II) Prepare groups separately.

R ¹⁰ in the above formula is R¹ described above or protected hydroxy methyl,

R 3 and R 각각 are each hydrogen or a lower alkyl protecting group,

The atomic arrangement at the carbon atom represented by (a) is (R) when R ¹⁰ is not hydrogen,

R ⁵ is a protected amino group,

R ²⁰ is R ² as described above or a corresponding protected form of hydroxy- (loweralkyl) or guindano- (loweralkyl),

The atomic arrangement at the carbon atom represented by (b) is (L).

The protected amino group represented by R ⁵ among the protected amino acids of the structural formula (III) as starting material may be any of the protected amino groups well known in the peptide chemistry art.

In a preferred embodiment of the preparation of the invention the amino group is protected with an aralkoxycarbonyl, in particular benzyloxycarbonyl, or tert-butoxycarbonyl group. However, amino groups can be protected with formyl, trityl, trifluoroacetyl, or 2 (biphenylyl) -isopropyloxycarbonyl groups, for example, or can be in the form of phthalimido groups.

The protecting group present in the protected hydroxy methyl group R ¹⁰ or the protected hydroxy (lower alkyl) group R ²⁰ can be any of the usual hydroxy protecting groups, for example aralkoxycarbonyl (eg benzyloxy Carbonyl), lower alkanoyl (eg acetyl, propionyl, etc.), aroyl (eg benzoyl), lower alkyl (eg tertiary butyl) or lower aralkyl (eg benzyl).

The condensation reaction of the compound of formula (II) with the protected α-amino acid of structure (III) can be carried out by methods known in peptamide chemistry. For example, it is carried out according to the mixed anhydride method, the azide method, the active ester method, the acid chloride method, the carbodiimide or the EEDQ (1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline) method. .

In one method, the compound of formula II can be condensed with the protected α-amino acid of formula III, wherein the carboxyl group is a mixed anhydride residue formed with an organic or inorganic acid. Suitably, the protected α-amino acid of formula (III) may be converted to tri (lower alkyl) amine (e.g., in an inert solvent such as tetrahydrofuran, 1,2-dimethoxyethane, dichloromethane, toluene, petroleum ether, etc.). Treated with a tertiary base such as triethylamine) or N-ethylmorpholine, and the resulting salts are reacted at a low temperature with a suitable chloroformate (e.g. lower alkyl formate such as ethyl chloroformate or isobutyl chloroformate). Next, the resulting mixture aqueous is suitably condensed with the compound of formula II in situ.

Alternatively, the compound of formula II is reacted with a protected α-amino acid of formula III, wherein the carboxy group is in the acid azide form.

This condensation reaction is preferably carried out at low temperature in the presence of an inert organic solvent such as dimethylformamide or ethyl acetate.

Alternatively, a compound of formula (II) may be prepared, wherein the carboxy group is of the active ester form (e.g. 4-nitrophenyl, 2,4,5 trichlorophenyl, or N-hydroxysuccinimide ester The condensation reaction is suitably carried out in an inert polysolvent such as aqueous dimethylformamide, or when both R 3 and R 중 in the compound of formula (II) are lower alkoxy, It is also suitable to carry out in the same lower alkanol.

Alternatively, the compound of formula II is condensed with the protected α-amino acid of formula III, wherein the carboxy group is in acid chloride form. This condensation reaction is preferably carried out at low temperature in the presence of a base.

In another method, the compound of formula II is condensed with the protected α-amino acid of formula III in the presence of carbodiimide (ie dicyclohexyl carbodiimide) or EEDQ. This condensation reaction can be carried out in the presence of an inert organic solvent (eg methylene chloride or lower alkanols such as methanol, ethanol, etc.) at room temperature or below room temperature.

Separation of the protecting group from the protecting group or the condensation product can be carried out by known methods. That is, the method practically used for separation of a protecting group or the method described in the literature is used.

For example, alkoxycarbonyl (eg benzyloxycarbonyl), secondary butoxycarbonyl or biphenylyl-isopropyloxycarbonyl groups can be separated by hydrolysis (eg with HBr in glacial acetic acid). process). Aralkoxycarbonyl (eg benzyloxycarbonyl) may be separated by hydrocracking (eg in the presence of pd / c or platinum hydroxide).

Quaternary butoxycarbonyl or 2-biphenylyl-isopropyloxycarbonyl groups can be separated using hydrogen chloride in dioxane. The trityl group can be separated by treating with acetic acid, and the phthalimido group can be converted to an amino group by decomposition with hydrazine. Lower alkyl protecting groups, denoted R and R, can be isolated by treatment with HBr in glacial acetic acid or by treatment with trimethyl chlorosilane or trimethylbromosilane followed by aqueous hydrolysis. If more than one protection group is present, the separation of these protections can be performed in one or several steps depending on the nature of the groups. However, it is preferable to use a protection group that can be separated in one step.

The compounds of formula (I) above are positive and pharmaceutically toxic with strong acids (ie hydrochloric acid, bromic acid, sulfuric acid, methanesulfonic acid, paratoluenesulfonic acid, etc.) and bases (ie HaOH, KOH, etc.) Forms nontoxic salts.

Starting materials of the formulas (II) and (III) are known compounds or can be prepared by known methods.

Peptide derivatives prepared according to the present invention have an antimicrobial activity against a wide range of Gram-positive and Gram-negative bacteria such as: Escherichia coli, Staphylococcus aureus, Hemolytic, Inpurenza, Modified bacteria, Young bacteria, Stool Streptococcus, Bacillus subtilis, Jutis bacteria, Pseudomonas aeruginosa, existing bacteria, etc.

In addition, the peptide derivatives of the present invention enhance the effectiveness of antibiotics, including penicillin and cephalosporin antibiotics and D-cycloserine. Antibiotics that are enhanced by the peptide derivatives of the present invention include amoxicillin, cepradine, cephalotin, cephalexin, calbenicillin, ampicillin, penicillin G, sulfenicillin, cefazoline, sepoxithin, rifampicin, [( R) 1- (2-furoyloxy) -3-methylbutyl] penicillin, (6R) -6-[[((hexahydro-1H-azin-1-yl) -methylene amino] penicillic acid, sephamandol, (Pivaloyloxy) methyl (6R) -6 [[((hexahydro-1H-azin-1-yl) methylene] amino] penicillane cephaloridine, cephaloglycine, peneticillin, methicillin, propi Silin, tikaracillin, amoxicillin, arginine salt, phosphonomycin, vancomycin and kanamycin.

The present invention also encompasses pharmaceutical preparations containing the aforementioned peptide derivatives, which may contain other antibiotics with additives as necessary.

Carriers used in the pharmaceutical preparations according to the present invention may be compatible with the peptide derivatives described above or with antibiotics, if any, and may be solid or liquid carrier materials suitable for administration.

The carrier material is an organic or inorganic carrier material suitable for enteral (eg oral) or parenteral administration. Carriers include water, gelatin, lactose, starch, magnesium, stearate, talc, vegetable oil, gum arabic, polyalkyleneglycol, jerry and the like.

The pharmaceutical preparations may be formulated in solid form (eg tablets, dragees, suppositories or capsules) or in liquid form (eg solutions, suspensions, emulsions). Pharmaceutical formulations may be used with conventional sterilization methods and may contain auxiliaries such as preservatives, stabilizers, wetting agents, emulsifiers, salts and buffers used to change the osmotic pressure. The range of pH when using buffers is known in the art. When the pharmaceutical formulation of the present invention contains a peptide derivative and an antibiotic, the weight ratio of the peptide derivative and the antibiotic is very wide.

In general, pharmaceutical preparations may comprise peptide derivatives and antibiotics in a weight ratio of 1: 100 to 100: 1, preferably 1:64 to 64: 1, but particularly preferred is a weight ratio of 1: 16-16: 16: 1. .

The daily dose of peptide derivatives, administered alone or in combination with antibiotics, varies widely depending on the type of peptide derivative, the type of antibiotic, the route of administration and the infection to be treated. By way of example, a daily dose of about 2000 mg to 400 mg when administered orally when administered with a peptide derivative alone is 800 to 2000 mg when administered parenterally. When the peptide derivative and the antibiotic are administered in combination, the daily dose for oral administration is about 750 to 1500 mg in the combined dose of the peptide derivative and the antibiotic, and the daily dose for parenteral administration is about 200 mg to 2000 mg.

The daily dose may be administered once or in divided doses and, as already mentioned, is tailored to the particular circumstances as determined by the prescribing physician in varying degrees depending on the individual's needs. The following examples illustrate the invention in detail.

Example 1

(A) Preparation of Starting Material

Dissolve 5.0 g (48 mmol) of L-α-aminobutyric acid in 24 ml of 2-N sodium hydroxide and dissolve the solution to 0 ° C. 12.3 g (72 mmol) of benzyl chloroformate and 18 ml (72 mmol) of 4-N sodium hydroxide are added alternately for 0.5 h with stirring, keeping the temperature at above 10 ° C., keeping the pH at about 11 and stirring. The mixture is slowly lowered to room temperature and stirred overnight. The mixture is slowly lowered at room temperature and stirred overnight. The mixture is extracted with 50 ml of dimethyl ether and the organic and aqueous phases are separated. 5N hydrochloric acid is added to the aqueous phase and acidified to pH 2 to obtain an oily mixture. The mixture is extracted twice with 75 ml of dimethyl ether. The extract is dehydrated over sodium sulfate and evaporated to give 9.6 g of colorless oil. This oil is dissolved in 10 ml of ethyl acetate and 100 ml of petroleum ether (boiling point 40 to 60 ° C.) is added to obtain an oily solution which is left at 0 ° C. The white crystalline material obtained was filtered and washed with petroleum ether to yield 8.40 g (73%) of N-benzyloxycarbonyl-L-a-aminobutyric acid having a melting point of 76 to 78 ° C.

(C = 1% in ethanol).

(B) process

(Iii) Dissolve 8.30 g (35 mmol) of N-benzyloxycarbonyl-L-α-aminobutyric acid and 4.02 g (35 mmol) of N-hydroxy succinimide in 75 ml of dimethoxyethane with stirring and the solution to 0 ° C. Cool. 7.94 g (38.5 mmol) of dicyclohexyl carbodiimide were added and the mixture was stirred at 0 ° C. for 2 hours and then left at 0 ° C. overnight. The byproducts of the solid are filtered off.

The filtrate is evaporated to yield a hard gum of N-benzyloxy carbonyl-L-α-aminobutyric acid N-hydroxysuccinimide ester. This is used without further purification.

(Ii) 4.38 g (35 mmol) of (1R) -1-aminoethylphosphonic acid is dissolved in a mixture of 40 ml of water, 7.07 g (70 mmol) of triethylamine and 40 ml of dimethylformamide while stirring. The resulting solution was cooled to 0 ° C. and 35 mmol of N-benzyloxycarbonyl-L-α-aminobutyric acid N-hydroxysuccinimide dissolved in 40 ml of dimethylformamide was added dropwise. The mixture is stirred at 0 ° C. and raised to room temperature with stirring overnight. The mixture is filtered to remove small amounts of solids. The filtrate was evaporated under an oil-pump vacuum and the residue was dissolved in a mixture of 30 ml of methanol and 10 ml of water and passed through an ion exchange resin column (BDH, Zerclit 225, SRC 13, RSO ₃ H: 150 g: regenerated in an acid cycle). Elute with solvent.

Collected in the acid eluate and evaporated to give a slightly gummy white solid. This solid is treated with 100 ml of diethyl ether and the diethyl ether is decanted and removed. The residual solid is dissolved in a mixture of 100 ml of methanol and 50 ml of water and the resulting solution is titrated with pH 4-4.5 with 4-N aqueous benzylamine (titrant: 8.0 ml, theoretical 8.75 ml).

The mixture solidifies upon reaching the end point of the titration.

The precipitate was filtered off, washed with water and dried to give: (1R) -1-[(N-benzyloxycarbonyl-L-α-aminobutyryl) amino] -ethylphosphonic acid having a melting point of 228 to 231 ° C (decomposition). 8.84 g of benzylamine salt are obtained.

(Iii) 45% HBr in which 8.7 g (19.3 mmol) of monobenzylamine salt of (11R) -1-[(N-benzyloxycarbonyl-L-α-aminobutyryl) amino] -ethylphosphonic acid was dissolved in glacial acetic acid 20 ml of a mixture of 8 ml of glacial acetic acid is stirred at room temperature for 6 hours. 100 ml of ether is added and the specimen is precipitated. This specimen was decanted by adding 100 ml of diethyl ether and dissolved in 100 ml of methanol. The solution is stirred and added twice with 5 ml of propylene oxide to pH 5 to obtain a white precipitate. The mixture was left overnight and the precipitate was filtered, washed with methanol and diethyl ether and dried to give crude (1R) -1- (L-α-aminobutyrylamino] -ethylphosphonic acid 4.05 having a melting point of 98 ° C. (decomposition). get g

Recrystallized from a mixture of 750 ml of water and 1500 ml of ethanol to obtain a white crystalline precipitate which was washed with ethanol, washed with diethyl ether and dried under vacuum to give (1R) -1- (L-) having a melting point of 296 to 298 ° C (decomposition). 3.58 g of α-aminobutyrylamino) -ethylphosphonic acid is obtained.

Example 2

(A) Preparation of Starting Material

5.0 g (42.5 mmol) of L-Norvaline are treated with benzylchloroformate and 10.2 g (60 mmol) sodium hydroxide as in Example 1 (A). The crude oil product is recrystallized from 10 ml of diethyl ether and 20 ml of petroleum ether (boiling point 40 ° to 60 ° C.) to give 8.2 g (77%) of N-benzyloxycarbonyl-L-norvaline having a melting point of 85 ° to 87 ° C. .

(B) process

(V) 8.1 g (32 mmol) N-benzyloxycarbonyl-L-norvaline, 3.68 g (32 mmol) N-hydroxysuccinimide, in the same manner as in Example 1 (B) (iii) ) And 7.21 g (35 mmol) of dicyclohexyl carbodiimide were treated with 25 ml of ethanol to obtain a white crystalline substance, and then 25 ml of petroleum ether (boiling point 40 to 60 ° C.) was added and filtered. 9.82 g of N-benzyloxycarbonyl-L-norvaline N-hydroxy succinimide ester can be used directly in the next step. Melting point of the purified sample is 95-97 ° C.

(Ii) 9.82 g (28 mmol) N-benzyloxycarbonyl-L-norvaline N-hydroxysuccinimide ester as described in Example 1 (B) (ii), and (1R) -1- (1R) -1-[(N-benzyloxycarbonyl) having a melting point of 225 to 230 ° C (decomposition) by using 37.5 g (30 mmol) of aminoethylphosphonic acid and treating with a mixture of 150 ml of methanol and 30 ml of water instead of diethyl ether 7.61 g of monobenzylamine of -L-norvaline) amino] -ethylphosphonic acid are obtained.

The filtrate was evaporated, the residue was treated with 100 ml of hot water, filtered and washed with ethanol and again with diethyl ether to give a second product, (1R) -1-[(decomposition) with a melting point of 231 to 234 ° C. (decomposition). 1.99 g of monobenzylamine salt of benzyloxycarbonyl-L-norvaline) amino] -ethylphosphonic acid are obtained.

인 (1R)-1-(L-노르발린아미노)-에틸포스폰산 3.54g(82%)을 수득한다.(Iii) the same procedure as in Example 1 (B) (iii), except for (1R) -1-[(N-benzyloxycarbonyl-L-norvaline) amino] -monophosphonic acid monobenzylamine Melting point 260-262 DEG C (decomposition) by recrystallization from a mixture of 80 ml of water and 160 ml of ethanol using 9.0 g (19 mmol) of salt:

3.54 g (82%) of phosphorus (1R) -1- (L-norvalineamino) -ethylphosphonic acid is obtained.

Example 3

(A) Preparation of Starting Material

5.0 g (38 mmol) of L-norleucine are treated with 9.7 g (57 mmol) of benzylchloroformate and sodium hydroxide according to the same method as in Example 1 (A). The product N-benzyloxycarbonyl-L-norleucine is crystallized and liberated in the form of an oil.

(B) process

(Iii) the same procedure as in Example 1 (B) (iii), except for N-benzyloxycarbonyl-L-norleucine (about 38 mmol), 4.38 g (38 mmol) of N-hydroxysuccinimide; N-benzyloxycarbonyl-L-norleucine N-hydroxy succinimide as a crystalline solid was treated with 8.65 g (42 mmol) of dicyclohexylcarbodiimide and the resulting crude oil product was treated with 100 ml of diethyl ether. 7.47 g of ester are obtained.

Melting point 82-84 ° C:

(Ii) Performing the same method as Example 1 (B) (ii), except for 7.40 g (20.5 mmol) of N-benzyloxycarbonyl-L-norleucine N-hydroxysuccinimide ester and (1R) -1 2.56 g (20.5 mmol) of aminoethyl phosphonic acid, methanol / water (5: 1) instead of methanol / water (3: 1) in the ion exchange step, and the resulting crude product was evaporated and treated with ether. 5.64 g of (1R) -1-[(N-benzyloxycarbonyl-L-norlosilyl) amino] -ethylphosphonic acid having a melting point of 192 to 194 ° C (decomposition), dissolved in 150 ml of hot water, and cooled. (74%) is obtained.

(Iii) Example 1 (B) (iii), but in the same manner as (1R) -1-[(N-benzyloxycarbonyl-L-norrocyl) amino] -ethylphosphonic acid 5.20 g (14.0) Mmol), and the obtained product was recrystallized from a mixture of 70 ml of water and 280 ml of ethanol to obtain 3.02 g of (1R) -1- (L-norroylamino) -ethylphosphonic acid having a melting point of 254 to 256 ° C (decomposition). 91%).

Example 4

(Iii) 17.4 g of N-benzyloxycarbonyl-L-norvaline N-hydroxy succinimide ester as in the same manner as in Example 1 (B) (ii) and Example 2 (B) (ii) (50 mmol) and 5.55 g (50 mmol) of aminomethyl phosphonic acid were treated with benzylamine in a mixture of 250 ml methanol and 50 ml to give 17.3 g of the crude monobenzylamine salt of the product (product) having a melting point of 195 to 198 ° C. Get The filtrate is evaporated and the residue is recrystallized from a mixture of methanol (80 ml) and water (20 ml) to give 2.19 g of pure product having a melting point of 180 to 185 ° C. (decomposition). Total yield: 19.5g (86%).

0.5 g of product (1) was recrystallized from 10 ml of hot water to obtain 0.39 g of monomethylamine salt of pure (N-benzyloxycarbonyl-L-norvaline) amino methylphosphonic acid having a melting point of 203 to 205 ° C (decomposition). .

(b) Performing the same method as Example 1 (B) (iii), using 18.9 g (42 mmol) of the monobenzylamine salt of (N-benzyloxy carbonyl-L-norvaline) -aminomethylphosphonic acid Recrystallization from a mixture of 80 ml of water and 160 ml of methanol yielded 6.65 g (75%) of L- (norvalineamino) methyl phosphonic acid having a melting point of 273 to 275 ° C (decomposition).

Example 5

N-benzyloxycarbonyl-L-norvaline (45.4 g, 0.181 mol) is stirred in methylene chloride solution (200 ml) and dimethyl aminomethylphosphonate hydrochloride (31.76 g, 0.181 mol) is added. The resulting suspension is cooled to −12 ° C. and anhydrous triethylamine (25.3 ml 0.181 mol) is added dropwise. At the end of the dropping, the cold mixture is stirred for 15 minutes and E, E, D, Q (56.8 g, 0.23 mol) dissolved in methylene dichloride (100 ml) is added rapidly. The mixture is stirred with cooling for 2 hours and stirred at room temperature for 1 week. The mixture is washed with water (100 ml) and again with 1N hydrochloric acid (4 times, 100 ml each). The total pickling is extracted with methylenedichloride (2 x 50 ml) and the combined organic solutions are washed again with water (100 ml) and finally with 15% KHCO 3 solution (3 x 100 ml) and then dehydrated over anhydrous sodium sulfate.

The solution is filtered and evaporated. The residual oil is redevaporated with benzene to give 78.13 g of residue. First crystallize. The oil is treated with anhydrous ether (200 ml). Very rapid crystallization spreads into solution.

The solution is refrigerated for 1 hour, the solid is filtered off, washed with ether and dried to constant weight under dryness to give 58.2 g of material having a melting point of 77 to 80 ° C. This is dissolved in hot ethyl acetate (200 ml) and the solution is filtered. To the cooled filtrate is added ether (200 ml).

The mixture is crystallized and refrigerated overnight, then the solid is filtered off, washed with ether and dried in vacuo to yield 50.64 g of a residue having a melting point of 82 to 84 ° C.

[(N-benzyloxycarbonyl-L-norvalyl) amino] dimethyl phosphate is stirred for 5 hours in 45% HBr dissolved in acetic acid (120 ml). At first, carbon dioxide is generated rapidly.

Ether (500 ml) is added to precipitate oil. The mixture is stirred for 40 minutes and left as it is. The ether is decanted off and the oil is washed twice with 500 ml each. Dissolve the oil in methanol (300 ml), stir the solution and add propylene oxide (40 ml). After about 5 minutes the product starts to crystallize. The mixture is adjusted to pH 4 and refrigerated overnight. Filter through solid, wash well with methanol and then ether. It is dried in vacuo to give 20.13 g of residue having a melting point of 228 to 289 ° C.

It is dissolved in hot water (200 ml) and filtered.

400 ml of ethanol is added to the filtrate and left to crystallize.

It was refrigerated for 2 hours, the solid was filtered, washed with ethanol, washed with ether again, dried over phosphorus pentoxide under vacuum, and 16.66 g of (L-norvalylamino) -methyl phosphonic acid having a melting point of 293 to 294 ° C (decomposition). To obtain.

Example 6

(a) L-nitro-homoarginine (2.33 g, 10 mmol, melting point 227-230 ° C. (decomposition)) was treated with 3.41 g (20 mmol) benzylchloroate and sodium hydroxide according to the method of Example 1 (A). do. The crude oily product is recrystallized from ethyl acetate (2 × 100 ml) instead of diethyl ether.

The crude oil phase product N-benzyloxycarbonyl-W-nitro-L-homoarginine obtained is dehydrated over sodium sulfate and evaporated to give a crude material which can be used in the next step without purification.

(b) Performing the same method as Example 1 (B) (iii), using dimethylformamide (50 ml) as a solvent, and crude N-benzyloxycarbonyl-W-nitro-L-homoarginine (obtained above About 4.2 g (about 10 mmol), 1.15 g (10 mmol) of N-hydroxysuccinimide and 2.27 g (11 mmol) of dicyclohexylcarbodiimide were used to remove 1.73 g of dicyclohexylene and the filtrate was Evaporation gives the crude product N-benzyloxycarbonyl-W-nitro-L-homoarginine N-hydroxysuccinimide ester on yellow oil, which is used in the next step without further purification.

(C) By the method of Example 1 (B) (ii), about 10 mmol of N-benzyloxy carbonyl-W-nitro-L-homoarginine N-hydroxysuccinimide ester (1R) -1-amino It is reacted with 1.25 g of ethylphosphonic acid and treated with ion exchange resin RSO ₃ H using 5: 1 instead of MeOH: H ₂ O 3: 1. The eluate was evaporated and partitioned into water (300 ml) and ethyl acetate (150 ml) to obtain an insoluble substance which was filtered and dried to yield 1.36 g of crude product (product 1, checked by TLC and NMR), having a melting point of 190 to 193 ° C. To obtain.

The filtrate solvent layer is separated, the aqueous layer is evaporated and dried and the residue is treated with acetone (50 ml) to give a white precipitate.

The precipitate was filtered off, washed with acetone and dried to give (1R) -1-[(N benzyloxycarbonyl-W-nitro-L-homoarginine) amino] -ethylforce having a melting point of 184 to 188 ° C (decomposition). 1.43 g of phonic acid (product 2) are obtained.

Product 1 and Product 2 are used in the next short without the need for further purification.

(d) The same procedure as in Example 1 (B) (iii), except for (1R) -1-[(N-benzyloxycarbonyl-W-nitro-L-homoalginyl) amino-ethylphosphonic acid ( (1R) -1- (LW-nitro homoal group having a melting point of 190 ° C. (decomposition) using 2.7 g (about 5.7 mmol) of the products 1 and 2) obtained above and recrystallized with a mixture of 15 ml of water and 150 ml of ethanol. 1.23 g of nile amino) -ethylphosphonic acid are obtained.

The filtrate was evaporated and recrystallized from water (150 ml) and ethanol (100 ml) to yield 0.14 g of product having a melting point of about 190 ° C. (decomposition).

Total yield: 1.37 g (71%)

(e) 1.2 g (3.5 mmol) of (1R) -1-LW-nitro-homoarginyl amino-ethylphosphonic acid prepared above was dissolved in 500 ml of water, and 0.35 g of 10% pd / C was added. This mixture is hydrogenated at room temperature and atmospheric pressure. The mixture is filtered to remove the catalyst and the liquor is evaporated. The solid residue was dissolved in 15 ml of cold water, the solution was filtered, and the solution was treated with 120 ml of ethanol to obtain a precipitate. 0.76 g of amino) -ethylphosphonic acid is obtained.

Example 7

(Iii) 45 mmol of N-benzyloxycarbonyl-L-α-aminobutyric acid N-hydroxysuccinimide ester and aminomethylphosph in the same manner as in Example 1 (B) (ii) dissolved in 40 ml dimethylformamide [(N-benzyloxycarbonyl-L-α-aminobutyryl) having a melting point of 185-195 ° C. (decomposition) using 44.4 g (40 mmol) of phonic acid and 4: 1 instead of methanol: water 2: 1. 14.1 g of monobenzylamine salt of amino] -methyl-phosphonic acid is obtained. The filtrate was evaporated and the residue was recrystallized from a mixture of 50 ml of water and 50 ml of methanol to yield 1.6 g of a white crystalline precipitate, a product having a melting point of 205 to 2.8 ° C. (decomposition).

(Ii) The same procedure as in Example 1 (B) (iii), except for mono ((N-benzyloxycarbonyl) -L-α-aminobutyryl) amino] -methyl-phosphonic acid, prepared above. Using 15.6 g (about 36 mmol) of benzylamine salt, 5.92 g of crude product having a melting point of 253 to 255 ° C. (decomposition) is obtained.

The mixture was recrystallized from 60 ml of water and 120 ml of ethanol to obtain 4.93 g of (L-α-aminobutyrylamino) -methylphosphonic acid having a melting point of 263 to 265 ° C (decomposition).

Example 8

(Iii) 2.52 g (7 mmol) N-benzyloxycarbonyl-L-norycin N-hydroxysuccinimide ester and 0.78 aminomethylphosphonic acid in the same manner as in Example 3 (B) (ii) Treat the acid eluate with benzylamine directly using g (7 mmol) and 2: 1 instead of methanol / water 5: 1 in the ion exchange step (evaporation, treatment with ether and subsequent dissolution in methanol / water are omitted) Agitated with 100 ml of acetone to yield 0.56 g of monobenzylamine salt of [(N-benzyloxycarbonyl-L-norroyl) amino] -methyl phosphonic acid having a melting point of 185-189 ° C. (decomposition). The solution is evaporated and treated with 50 ml of diethyl ether to further obtain 1.9 g of product having a melting point of 170 ° to 180 ° C. (decomposition).

(Ii) 2.40 g of monobenzylamine salt of [(N-benzyloxycarbonyl-L-norroyl) amino] -methyl-phosphonic acid, prepared in the same manner as in Example 1 (B) (iii) From about 5.2 mmol, the two products are used to obtain 1.04 g of crude product having a melting point of 263 to 266 ° C. (decomposition).

The mixture was recrystallized from 20 ml of water and 120 ml of ethanol to give 0.88 g of (L-norroylamino) -methylphosphonic acid having a melting point of 272 to 274 ° C (decomposition).

The following describes a typical pharmaceutical formulation containing the peptide derivative of the present invention.

Formulation Example A

Parenteral preparations containing the following active ingredients may be formulated.

Claims (1)

A process for preparing a peptide derivative of formula (I) characterized by condensing a compound of formula (II) with a protected α-amino acid of structure (III) and separating the groups present in the protecting group or condensation product .

In the above structural formula R¹ is hydrogen, methyl or hydroxymethyl, or mono-, di- or trihalomethyl, R 2 is lower alkyl, hydroxy- (lower alkyl) or guanidino (lower alkyl), which is not a characteristic group of α-amino acids of the type normally found in proteins, The arrangement at the carbon atom represented by (a) is (R) when R¹ is not hydrogen, The arrangement at the carbon atom represented by (b) is (L), R ¹⁰ is R 1 described above, or protected hydroxy methyl, R³ and R⁴ are hydrogen or lower alkyl protecting group, The arrangement at the carbon atom represented by (a) in formula (II) is (R) when R ¹⁰ is not hydrogen, R ²⁰ is R ² as described above or a protected hydroxy- (lower alkyl) or guanidino- (lower alkyl) group R ⁵ is a protected amino group and the arrangement at the carbon atom represented by (b) in formula (III) is (L).