CS202564B2 - Process for preparing derivatives of 6-/d-2-amino-2-phenylacetamido/penicilanic acid - Google Patents

Abstract

The compounds of the formula I, in which the substituents have the meaning mentioned in Claim 1, are obtained by condensaton of 6-(D-2-amino-2-phenylacetamido)penicillanic acid having an optionally protected acid group with an N-protected amino acid of the formula HOOC-CH(NHZ)-CH2COR<3'> or HOOC-CH2-CH(NHZ)-COR<3'>. The novel penicillins obtained in this manner are antibacterially active and are suitable as animal feed additives and for the control of infectious diseases caused by Gram-positive or Gram-negative bacteria. <IMAGE>

Description

The invention relates to a process for the preparation of 6- [D-2-acylamido-2-phenylacetamido) penicillanic acid of the formula I

With ^CH 3 Q CQOH (1) in which

R1 represents a hydrogen atom or ¹ hydroxyl group,

R ² represents a group of formula -CO-CH (NH2 J - CH2 COR ³ or -COCH 2 CH (NH 2) - COR ³ and R 3 is selected from the group consisting of hydroxyl, lower alkylamino, di-lower alkylamino, lower alkoxy and hydroxy-lower an alkylamino group or, in the case where R 1 represents a hydroxyl group, R 3 represents an amino group.

The terms "lower alkyl" and "lower alkoxy" in the specification. The invention and the definition of the invention are to be understood as branched or unbranched alkyl and alkoxy groups having 1 to 6 carbon atoms.

U.S. Patent Nos. 3,268,513 and 3,340,252 disclose that 6- (D-2- (DL-2-aminoglutaramido) -2-phenylacetamido) -pentanoic acid is prepared by the reaction of 6- (D-2-amino-2-phenylacetamido) -penicillanic acid DL-glutamine, and that said glutarimide derivative has an antimicrobial effect against gram-positive and gram-negative bacteria. On the other hand, German patent application DOS '1 934 783 discloses that 6- (D-2- (L-2-) acid). 4'-imine-2-phenylacetamido] penicillanic acid is prepared by condensation with diphenylisopropyloxycarbonyl-L-asparagine followed by deprotection.

The present invention provides novel penicillins that are useful as antibacterial agents, as feed supplements in animal nutrition, and as chemotherapeutic agents for poultry and mammals, including humans, in the treatment of infectious diseases caused by Gram-positive and Gram-negative bacteria. In particular, the penicillanic acid compound of the formula I according to the invention shows a significant antimicrobial action against a wide variety of microorganisms and can be effectively absorbed by either oral or parenteral administration to humans and animals. In particular, penicillanic acid, a compound of penicillanic acid I, exhibits a significant antimicrobial action against bacteria belonging to the genera Pseudomonas and Staphylococcus. For example, 6- [D-2-amino-3-N-methylcarbamoylpropionamido] -2-p-hydroxyphenylacetamido] penicillanic acid exhibits minimal inhibitory concentrations [Μ. I. C.j. Agar dilution method, on heart broth agar, about 12.5 µg / ml against Pseudomonos aeruginosa Аз. On the other hand Μ. I. C. sulbenicillin [i. disodium salts of 6- (D-2-sulfonyl-2-phenylacetamido) penicillanic acid, 6- [D-2- (DL-2-aminoglutaramido) -2-phenylacetamido] penicillanic acid and 6- [D- 2- (L-2-amino-3-carbamoylpropionamido] -2-phenylacetamido] penicillanic is 12.5, 50 and more than 100 [mu] g / ml against said microorganism. The N-methylcarbamoylpropionamido) -2-hydroxyphenylacetamido] penicillanic compound according to the invention against Staphylococcus aureus ATCC 6538 for St. epidermis 10 131R is also about 2-4 times higher than the effect of Sulbenicillin and 6- [D-2- (L-amino- The penicillanic acid compound I according to the invention can furthermore have a significant antimicrobial action against Gram-positive and gram-negative bacteria, for example belonging to the genera Streptococcus, Escherichia, Klebsiella, Salmonella and Bacillus. distinctly low For example, when 6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid is administered to mice at a dose of 10,000 mg / kg, none of the mice will die in 14 days after application.

The penicillanic acid compound I of the invention may find pharmaceutical use either in the form of the free acid or a salt thereof. Physiologically acceptable salts of the compounds I are, for example, non-toxic salts of metals such as sodium, potassium, calcium and aluminum, ammonium and substituted ammonium salts, for example salts of non-toxic amines such as trialkylamines including triethylamine, procaine, dibenzylamine. N-benzyl-β-phenethylamine, L-efenamine, Ν, Ν'-dibenzylethylenediamine, dehydroabietylamine, N, N'-bis-dehydroabietylethylenediamine and other amines used to prepare salts with benzylpenicillin. The penicillanic acid compound I is very water soluble and can be administered either orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously). The daily dose of Compound I may be from 0.25 to about 20 g, in particular 0.5 to 10 g, and especially about up to about 4 g for an adult patient. Compound I may further be used in the form of a pharmaceutical composition comprising the same compound in conjunction with or in admixture with a pharmaceutical diluent suitable for enteral or parenteral administration. Suitable diluents are, for example, gelatin, lactose, glucose, sodium chloride, starch, magnesium stearate, talc, vegetable oil and other known pharmaceutical diluents. The pharmaceutical preparations may be in solid form as tablets, dragees, pills or capsules or in liquid form as solutions, suspensions or emulsions. They may be sterilized and / or may further contain adjuvants such as stabilizing, wetting and emulsifying agents.

The penicillanic acid compound I of the present invention can be prepared by condensation of 6- (D-2-amino-2-phenylacetamido) penicillanic acid of formula II

R ¹ has the abovementioned meaning and

Y ¹ represents a hydrogen atom or a protecting group, which is a benzyl, phthalimidomethyl, 2,2,2-trichloroethyl or phenacyl group, with an N-protected amino acid of the general formula III

W — ОН (III), in which

W represents a group of formula -COCH (NHZ) - CH2 COR ³ or -COCH 2 CH (NHZ) optionally -COR ³ where R ³ in compound I represents a hydroxy group, W may be a group of the formula -COCH (NHZ) ^2, or Y -CH2COO —COCH2 — CH (NH2) —COOY ²

R ³ has the abovementioned meaning,

Y ² denotes a benzyl group and

Z is a protecting group which is O-nitrophenylsulfenyl, benzyloxycarbonyl or 1-methyl-2-methoxycarbonylvinyl in the presence of diphenylphosphoric azide and at a temperature of 20 to -20 degrees Celsius in an inert solvent or with a reactive derivative of a compound of formula III at from 40 to -30 ° C in an inert solvent to give 6- (D-2-acylamido-2-phenylacetamido) penicillanic acid of formula IV

CH-CONH

in which

R ¹ , Y ¹ and W are as defined above, after which the protecting group (s) are removed from compound IV.

Removal of the protecting group or groups from compound IV when Z is an o-nitrophenylsulfenyl group is carried out by (i) treatment with two to three molar equivalents of thioamide at 40 to 0 ° C, (i) when Z is benzyloxycarbonyl and / or a Y ¹ or Y ² benzyl group by shaking in a hydrogen atmosphere in the presence of palladium on VASO or palladium on carbon or palladium black at 40 to 0 ° C; (iii) when Z is a 1-methyl-2-methoxycarbonylvinyl group an aqueous solution of hydrohalic acid, (i) when Y ¹ is a phenacyl or phthalimidomethyl group, by treatment with an alkali metal salt of a thiol compound or thiophenol at a temperature of-10 to 10 ° C, (v) when Y ¹ is 2, 2,2-trichloroethyl group by reduction with formic acid and zinc dust at a temperature of 10 to -5 ° C.

The penicillanic acid compound II in which Y ¹ represents a hydrogen atom may be used in the reactions of the invention in the form of the free acid or a salt thereof. Examples of suitable said salts are alkali metal salts such as sodium and potassium salts and lower trialkylamine salts such as triethylamine and trimethylamine salts.

The condensation reaction of the compound of penicillanic acid II with amino acids III is readily feasible. For example, said condensation reaction is preferably carried out in the presence of diphenylphosphoric azide [i. №РО (ОСбН5) 2] in solvent. The reaction is suitably carried out at temperatures of from -20 to 20 ° C, in particular from -10 to 5 ° C. The reaction is particularly preferably carried out in the presence of an organic tertiary amine such as trimethylamine and triethylamine. Suitable reaction solvents are dimethyl formamide, dimethylsulfoxide and dichloromethane. The acid III may optionally be converted into its reactive derivatives prior to the condensation reaction mentioned above. Suitable examples of reactive derivatives of these acids are the corresponding mixed anhydrides (e.g. lower alkoxycarbonyl esters such as ethoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl and tert-amyloxycarbonyl esters, lower alkanoyl esters such as pivaloyl esters), reactive esters and nitro esters (e.g. hydroxysuccinimide) and azide. Mixed anhydrides of acid III can be prepared by conventional methods, for example by reaction of these acids with a lower alkoxycarbonyl halide (e.g. bromide or chloride) or a lower alkanoyl halide (e.g. bromide or chloride) at -40 to 5 ° C in the presence of an acid binder in a solvent . Preferred solvents are chloroform, dichloromethane, dimethylsulfoxide and dimethylformamide, examples of acid binders are N, N-dimethylaniline, N-methylmorpholine, pyridine etc. On the other hand, active esters of acid III can be prepared by reaction of this acid with p-hydroxyphenol or N -hydroxysuccinimide at -10 to 30 ° C in the presence of dehydrating agents (e.g. tetrahydrofuran, dioxane or diglyme). Azides can be prepared by reacting the hydrazides of these acids with sodium nitrite in a dilute mineral acid (e.g., sulfuric acid). The condensation reaction of the penicillanic acid compound II with the reactive amino acid derivative III thus obtained is carried out at a temperature of 40 to -30 ° C, in particular at a temperature of 5 to -15 degrees Celsius, in a solvent. Suitable reaction solvents are dimethylformamide, chloroform, tetrahydrofuran and methylene chloride.

Removal of the protecting group or groups from 6- (D-2-acylamido-2-phenylacetamido) penicillanic acid IV can be carried out by suitable methods depending on the type of protecting group or groups used. For example, when o-nitrophenylsulfenyl groups are used as a protecting group Z, said group can be removed by reacting compound IV with about 2 to 3 equivalents of thioamide in a solvent. Examples of suitable thioamide are thiacetamide, thiobenzamide, thiourea and 2-mercapto-5-methyl-1,3,5-triadiazole. The reaction is preferably carried out at temperatures from 40 to 0 ° C, in particular at temperatures from 20 to 10 ° C. Ethyl ether, tetrahydrofuran, dioxane, lower alkanol (e.g. methanol, ethanol) or a mixture of these solvents are suitable reaction solvents. If the protecting group Z is a benzyloxycarbonyl group and / or each of the protecting groups Y ¹ and Y ² is a benzyl group, removal of said protecting group or group may be accomplished - COCH (NH 2) CH 2 COOY 2 or (wherein the pin shaking compound IV with hydrogen gas in The catalytic hydrogenation is preferably carried out at a temperature of from 40 to 0 ° C, in particular at 20 to 10 ° C under normal pressure, Examples of preferred catalysts are palladium on barium carbonate, palladium on activated carbon and palladium black. If the protecting group Z is 1m, ethyl-2- (lower Jalkoxycarbonylvinyl), said group may be removed by treatment with dilute mineral acids (e.g. 0.5 to 1% hydrochloric acid) at a temperature of from 5 to 5%. 10 ° C in a solvent (e.g., methyl ethyl ketone, methyl isobutyl ketone) If phenacyl or phthalimidomethyl groups are used as protecting groups X ¹ , said group may be removed by reaction with an alkali metal salt of the thiol compound or thiophenol. This reaction is preferably carried out at a temperature of -10 to 10 ° C in a solvent such as tetrahydrofuran or isopropyl alcohol. The 2-ethylhexyl ester of 2-mercaptoacetic acid is preferably used as the thiol compound. If the protecting group Y ^{1 is a} 2,2,2-chloroethyl group, this group is removed by reducing the compound IV with 90% formic acid and zinc dust by conventional means. The reaction is preferably carried out at a temperature of 10 to -5 ° C.

The starting compound (III) according to the invention can be obtained by methods known per se. For example, an N-protected amino acid III in which W represents a group of formulas —COCH (NH2J — CH2COR3 or —COCH22NHZJ - COR3 (wherein R3 is as defined above) is prepared by reacting an amino acid of formula R3COCCH — CH (NH'2 ^! ) - COOH or

R 3 COCH (NH 2) —CH 2 COOH wherein

R ³ is as defined above, with a compound of formula

Z - X in which

X represents a halogen atom and

Z is as defined above.

The reaction may be carried out preferably at a temperature of 0 to 40 ° C in the presence of an acid-binding agent (e.g. sodium carbonate, potassium carbonate sodium hydroxide) in a solvent (e.g. water, tetrahydrofuran, dioxane or mixtures thereof).

The starting compound of formula (III) wherein W is a group of formulas (COCH 22 NH ² ) -COOY ²

Y 2 and W have the meanings given above) can be prepared by reaction. amino acids of general formula

Y2OCO — CH2CH (NH2) —COOH or

Y 2 OC-CH (NH 2) -CH 2 COOH wherein

Y ² is as defined above, with a compound of formula

Z — X in which

Z and X have the same meaning as previously described.

The practical examples and preferred embodiments of the invention are illustrated by the following examples.

He did

1) A solution of 1.5 g (11.8 mmol) of DL-2-amino-3-N-methylcarbamoylpropionic acid hydrochloride (i.e. DL-N'-methylasparagine hydrochloride) in 16 ml of water is neutralized with potassium carbonate and added thereto. 5 ml of tetrahydrofuran are added. To the solution was added 2.5 g of o-nitrophenylsulfenyl chloride at 5-10 ° C. The reaction mixture was stirred at this temperature for 2 hours. During the reaction, the mixture was kept slightly alkaline with potassium carbonate (pH 8). Water (10 ml) was added to the reaction mixture and the insoluble material was filtered off. The filtrate is washed twice with ethyl acetate, acidified with citric acid and then extracted three times with 50 ml of ethyl acetate. The ethyl acetate extracts were washed with water, dried and evaporated to remove the solvent. 2.8 g of DL-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoyl- are obtained. 151 DEG-152 DEG.

2) 1.5 g (5 mmol) of DL-2- (o-nitrophenylsulfenylamino) -3-N-methyl-4-carbonyl-propionic acid are dissolved in 30 ml of tetrahydrofuran. To the solution was added 1.14 g (5.5 mmol) of dicyclohexylcarbodiimide and 632 mg (5.5 mmol) of N-hydroxysuccinimide at 0-5 ° C and the mixture was stirred at this temperature for 16 hours. Insoluble materials are filtered off. The solvent is then removed by evaporating the filtrate at 20 degrees Celsius under reduced pressure. and the crystalline precipitate thus obtained is washed with a mixture of benzene and ether. 1.2 g of N- [DL-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropoxy] succinimide are obtained in crystalline form.

M.p. 133-135 ° C.

3) 450 mg (1.13 mmol) of N- [DL-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionyloxy] succinimide and 6.05 mg (1.3 mmol) of triethylamine salt of 6- [D- -2-Amino-2- (p-hydroxyphenyl) acetamido] penicillanic is dissolved in a mixture of 10 ml of chloroform and 15 ml of dimethylformamide • The solution is stirred at 0-5 ° C for 16 hours. The solution was evaporated at about 30 ° C under reduced pressure to remove the solvent, ether was added to the residue, and the precipitate thus obtained was collected by filtration to give 700 mg of 6- [D-2- (DL) acid as a yellow caramel. -2- (4-nitrophenylsulphenylamino) -3-N-methylcarbamoylpropionamido) -2-p-hydroixylphenylacetamido] penicilane.

Infrared absorption spectrum:

In max ° ^1: 3270, ¹⁷⁷⁵ > ¹⁷²⁰ ' ¹⁶⁴⁵ cm ⁴

4) 710 mg (1.1 mmol) of 6- [D-2- (DL-2-omtrophenylsulfenylamino) -3- N -methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid and 450 mg (3.3 mmol) of thiobenzamide is dissolved in a mixture of 20 ml of ethanol and 5 ml of tetrahydrofuran. The solution is stirred at normal temperature for 30 minutes. The reaction mixture was evaporated at about 30 ° C under reduced pressure to remove the solvent. The residue thus obtained is stirred with 20 ml of tetrahydrofuran and collected by filtration. The precipitates thus obtained are dissolved in 10 ml of aqueous tetrahydrofuran. To this was added 30 ml of ethyl acetate and the aqueous layer was separated. After washing twice with a mixture of tetrahydrofuran and ethyl acetate, the aqueous layer is freeze-dried. 450 mg of 6- [D-2- (DL-2-amino-3-N, N, 4 ', 3', 4 ', 4'-carbam (4, 3', 4 ', 4'-pionic acid) -2-p-hydroxyphenylacetamido) are obtained. penicilane in the form of a colorless powder.

M.p. 159-198 ° C (dec.). Infrared absorption spectrum:

ks ^ ¹ x ³² 8 ^0, 1760, 1650, 1595 cm ^1st

Thin layer chromatography:

Rf = 0.43 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)) ·

Infrared absorption spectrum of potassium salt:

h: 3300, 1760, 1650, 1595 cm ⁴ .

Example 2

1) 3 g of D-2-amino-hydrochloride (i.e., D-N-methyl-methylparagine hydrochloride) 5 g of o-nitrophenylsulfenyl chloride, 30 g of water, 10 g ml of tetrahydrofuran and 12 g of potassium carbonate were treated as described in Example 1 - 1). 5 g of D-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionic acid are obtained.

Melting point 134-136 ° C.

2) 1.57 g (5.2 mmol) of D-2- (p-nitrophenylsulphenylamino) -β-β-β-β-imidazole; ¹ Bamoylpropionic acid, 1.3 g dicyclohexylcarbodiimide, 788 mg N-hydroxysuccinimide and 15 ml tetrahydrofuran were treated in the same manner as described in the example.

1—2). 1.8 g of N- [D-2- (o-nitrophenylsulfenylamino) -3-N-methyl-1-carbamoylpropionyloxy] succinimide are obtained.

Melting point 126-128 ° C.

3) 800 mg (2 mmol) of N- [D-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionyloxy] succinimide and 932 mg (2 mmol) of the triethylamine salt of N- [D-2-amino-2- (p) Hydroxyphenylene acetamido-1-penicillanic acid is dissolved in 15 ml of dimethylformamide and stirred at 0 to 5 degrees Celsius for 16 hours, after which the solution is worked up as described in Example 1-3. 1.15 g of 6- [D-2- (D-2-o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionamido -) - 2-p-hydroxyphenylacetamide o] penicillanic acid are obtained in the form of a yellow powder.

Melting point 165-167 ° C (dec.).

4) 1.10 g (1.7 mmol) of 6- [D-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid and 830 mg (6.0 mmol) of thiobenzamide is dissolved in a mixture of 15 ml of methanol and 5 ml of tetrahydrofuran. The solution was stirred at normal temperature for 45 minutes. Thereafter, the reaction mixture was worked up in the same manner as described in Example 1-4). 700 mg of 6- [D-2- (D-amino-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid are obtained in the form of a colorless powder.

Mp 198-201 ° C (dec.).

Infrared absorption spectrum:

^In n ^1: 3280, 1760, 1660 cm ⁴ ,

Thin layer chromatography:

R f = 0.43 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Infrared absorption spectrum of potassium 3300 ^, 1760 ^, 1660 ^, 1600 cm ^-1 .

Common salts:

nujo ₁ ⁱⁿ max.

Example

1) 4.5 g [36 mmol) of acid hydrochloride. L-2-Amino-2-N-methylcarbamoylpropionic acid (i.e. L-N'-methylasparaffin hydrochloride), 7.6 g of o-nitrophenylsulfenyl chloride, 40 g of water, 20 ml of tetrahydrofuran and 18 g of potassium carbonate are treated as described above. in Example 1-1).

7 g of L-2- (o-isophenylsulfenylamine) -3-N-methylcarbamoyl propionic acid are obtained.

M.p. 132-135 ° C.

2) 3.2 g (11 mmol) of L-2- (o-nitrophenylsulphenylamino) -3-N-methylcarbamoylpropionic acid, 2.6 g of dicyclohexylcarbodiimide, 1.6 g of N-hydroxysuccinimide and 35 ml of tetrahydrofuran are treated. as in Example 1-2). 3.5 g of N- [L-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionyloxy] succinimide are obtained.

127 DEG-129 DEG.

3) 792 · mg (2 mmol) of N- [L-2- (o-nitrophinylsulfenylamino) -3-N-methylcarbamoylpropionyloxy] succinimide and 932 mg (2 mmol) of triethylamine salt of 6- [D-2-amino-2- (p- hydroxyphenyl) acetamido] penicillanic is dissolved in 15 ml of dimethylformamide and stirred at 0-5 ac for 16 hours, after which the solution is worked up as described in Example 1-3. 1.05 g of 6- [D-2- (L-2-p-hydroxyphenyl) acetamido] penicillanic acid is obtained in the form of a caramel.

Infrared absorption spectrum:

ΐ ^: 3260 1775, 1720, 1635 cm ^-1 .

4) 1.0 g (1.55 mmol) of 6- [D-2- (L-2- / o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionamido [2-p] hydroxyphenyl] acetamido-penicillanic acid and 740 mg (5.4 mmol) of thiobenzamide was dissolved in a mixture of 40 ml of methanol and 10 ml of tetrahydrofuran, and the reaction mixture was stirred at room temperature for 40 minutes, after which the reaction solution was worked up as described in Examples 1-4. ). 620 mg of 6- [D-2- (L-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid are obtained.

Mp 197-200 ° C (dec.).

Infrared absorption spectrum:

ⁱⁿ mma? ^1: 3250, 40, 1650 cm -1.

Thin layer chromatography:

R f = 0.43 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Example 4

1) 2.10 g (10 mmol) of D-2-amino-3-N-isopropylcarbamoylpropionic acid (i.e. D-N'-isopropylasparagine), 2.2 g of o-nitrophenylsulfenyl chloride, 20 ml of water, 20 ml of tetrahydrofuran and 1.0 g of potassium carbonate was treated as in Example 1 - 1).

2.3 g of D-2-o-isopropylphenylsulfenylamino) -3-N-isopropylcarbamoylpropionic acid are obtained.

M.p. 146-147 ° C (dec.).

2) 981 mg (3 mmol) of D-2- (o-nitrophenylsulfenylamino) -3-N-isopropylcarbamoylpropionic acid, 639 mg of dicyclohexylcarbodiimide, 356 mg of N-hydroxysuccinimide and 20 ml of tetrahydrofuran were treated as described in Example 1-2. ). 1.13 g of N- [D-2- (o-nitro-phenyl) -sulphenylamino] -3-N-isopropylcarbamoyl-propionyloxy-succinimide are obtained in crystalline form. Mp 144-145 ° C (dec.).

3) 699 mg (1.5 mmol) of triethylamine salt of 6- [D-2-amino-2- (p-hydroxyphenyl) acetamido] penicillanic acid are dissolved in 10 ml of dimethylformamide at a temperature of 3-5 ° C. 636 mg (1.5 mmol) of N- [D-2- (o-nitrophenylsulfenylamino) -3-N-isopropylcarbamoylpropionyloxy] succinimide are added and the mixture is stirred at this temperature for 2 hours. To 50 ml of ice-water, then the aqueous mixture is washed twice with 30 ml of ethyl acetate, acidified with citric acid and extracted three times with 30 ml of ethyl acetate, the combined extracts are washed three times with 20 ml of water, dried and evaporated under reduced pressure at less than 25 ° C to remove the solvent, gave 950 mg of 6- [D-2- (D-2- (o-nitrophenylsulfenylamino) -3-N-isopropylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid in crystalline form.

Mp 152-154 ° C (dec.).

Infrared absorption spectrum:

^V max ^1: 3275 '1780' 1730, 1640,

1620 cm -1.

4) 920 mg (1.36 mmol) of 6- [D-2- (D-2-t-enylsullenylamino) -3-N-isopropylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid are dissolved in a mixture of 20 · Ml of ethanol and 5 ml of tetrahydrofuran. To the solution is added 616 mg (4.5 mmol) of thiobenzamide at normal temperature and the solution is stirred at this temperature for 1 hour. The reaction mixture was evaporated under reduced pressure at a temperature below 25 ° C to remove the solvent. Ether was added to the residue and a crystalline precipitate was collected. After washing with tetrahydrofuran, the precipitate is dissolved in 50 ml of water. The aqueous solution was then washed twice with a 3: 1 mixture of tetrahydrofuran and ethyl acetate, washed with ether and lyophilized. Ether was added to the powdered lyophilizate and a crystalline precipitate was obtained by filtration. 620 mg of 6- [D-2- (D-2-amino-3-N-isopropylcarbamoylpropionamido) -2- (4-hydroxyphenylacetamide) penicillanic acid in crystalline form are obtained.

Melting point 198-200 ° C (dec.).

Infrared absorption spectrum:

νmax x ¹ : 3250, 1760, 1650 cm ^-1 .

Thin layer chromatography:

_Rf = 0.38 [silica gel plate, solvent: n-butanol-acetic acid (in the ratio 4: 1: 1)].

Example 5

1) 2.5 g (11 mmol) of D-2-amino-3-Nn-butylcarbamoylpropionic acid hydrochloride (ie D-N'-butylasparagine hydrochloride), 2.2 g of o-nitrophenylsulfenyl chloride, 25 ml of water, 25 ml of tetrahydrofuran and 1.2 g of sodium hydroxide are reacted under the conditions described in Example 1-1. 2.6 g of D-2- (o-nitrophenylsulfenylamino) -3-N-n-butylcarbamoylpropionic acid are obtained.

M.p. 143-144 ° C (dec.).

2) 1.023 g (3 mmol) of D- 2- (o-nitrophenylsulfenylamino) -3-N-butylcarbamoylpropionic acid, 639 mg (3.1 mmol) of dicyclohexylcarbodiimide, 356 mg (3.1 mmol) of N-hydroxysuccininide and 20 ml of tetrahydrofuran The reactions were carried out under the conditions described in Example 1-2. 1.04 g of N- [D-2- (o-nitrophenylsulfenylamino) -3-N-n-butylcarbamoylpropionyloxy] succinimide is obtained.

135 DEG-136 DEG C. (dec.).

3) 699 mg (1.5 mmol) of triethylamine salt of 6- [D-2-amino-2- (p-hydroxyphenyl) acetamido] penicillanic acid, 657 mg (1.5 mmol) of N- [D-2- ( o-nitrophenylsulfenylamino) -3-N-butylcarbamoylpropionyloxy] succinimide and 10 ml of dimethylformamide were reacted in the same manner as in Example 4-3). 1010 mg of 6- [D-2- (o-nitrophenylsulphenylamino) -3-N-n-butylcarbamoylpropionamido) -2-p-hydroxyphenylacetamide] penicillanic acid is obtained in the form of a caramel.

Infrared absorption spectrum:

h 3275, 1770, 1725, 1640 cm ¹ .

4) 980 mg (1.42 mmol) of 6- [D-2- (D-2- (o-nitrophenylsulfenylamino) -3-N-butylcarbamoylpropionamido) -2-p-hydroxyphenylacetamide] penicillanic acid, 616 mg (4) (5 mmol) of thiobenzamide and 15 ml of ethanol were reacted as described in Example 4- (4). 570 mg of 6- [D-2- (D-2-amino-3-N-n-butylcarbamoylpropionamide) -penicillanic acid is obtained in crystalline form.

Mp 188-191 ° C (dec.).

Infrared absorption spectrum:

nujol ^p max.

: 3270, 1760, 1650 cm ¹ .

Thin layer chromatography:

_Rf = 0.45 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Example 6

1) 2.7 g (11 mmol) of D-2-amino-3-Nn-hexylcarbamoylpropionic acid hydrochloride (ie D-N'-hexylasparagine hydrochloride), 2.2 g of o-nitrophenylsulfenyl chloride, 30 ml of water, 25 ml of tetrahydrofuran and 1.2 g of sodium hydroxide were reacted as described in Example 1-1. 2.8 g of D-2- (o-nitrophenylsulphenylamino) -3-N-n-hexylcarbamoylpropionic acid are obtained.

115 DEG-116 DEG C. (dec.).

2) 1.11 g (3 mmol) of D-2- (o-nitrophenylsulfenylamino) -3-Nnh, exylcarbamoylpropionic acid, 639 mg (3.11 mmol) of dicyclohexylcarbodiimide, 356 mg of N-hydroxysuccinimide and 20 ml of tetrahydrofuran are reacted as described in Example 1-2). 1.23 g of N-n-hexylcarbamoylpropionyloxy] succinimide are obtained.

128 DEG-129 DEG C. (decomposition).

3) 699 mg (1.5 µmol) of triethylamine salt of 6- [D-2-amino-2- (p-hydroxyphenyl) acetamido] penicillanic acid, 699 mg (1.5 mmol) of N- [D-2- (o nitrophenylsulfenylamino) -3-N-hexylcarbamoylpropionyloxy] succinimide and 10 ml of dimethylformamide were reacted as described in Example 4-3). 980 mg of 6- acid is obtained.

- [D-2- (o-nitrophenylsulfenylamino) -3-N-n-hexylcarbamoylpropionamido] -2-p-hydroxyphenylacetamido] penicillanic acid as a caramel.

Infrared absorption spectrum:

vm ^ x. ^{1 1} 3275, 1770, 1725, 1635 cm ^1st

4) 714 mg (1 mmol) of 6- [D-2- (o-nitrophenylsulfenylamino) -3-N-hexylcarbamoylpropionamido-2-p-hydroxyphenylacetamido] penicillanic acid, 472 mg (3.5 mmol) of thiobenzamide and 10 ml of ethanol are reacted as described in Example 4-4). 390 mg of 6- [D-2-] acid are obtained.

- (D-2-amino-3-N-n-hexylcarbamoylpropionamido) -penicillanic acid in crystalline form.

Infrared absorption spectrum:

in ^ ° ^1: 3250, 1770, 1655 cm ^1st

Thin layer chromatography:

Rf = 0.53 (silica gel plate, solvent: n-butanol-acetic acid-water 4: 1: 1).

20'2564

Example 7

1) 5 g (33 mmol) of D-2-amino-3-carbamoylpropionic acid (ie D-asparagine),

8.5 g. (O-nitroeenylsulphenyl chloride, 40 ml of water, 20 ml of tetrahydrolurane, and 5.5 g of potassium carbonate were reacted as in Example 1-1). 6.7 g of D-2- (o-nitrolenylsulenylamino) -3-carbamoylpropionic acid are obtained.

163 DEG C. (dec.).

2) 2.85 g (10 mmol) of D-2- (o-nitrolenylsulphenylamino) -3-carbamoylpropionic acid, 2.28 g of dicyclohexylcarbodiimide, 1.27 g of N-hydroxysuccinimide and 50 ml of tetrahydrolurane are reacted as described above. in Example 1-2). G of N- [D-2- (o-nitrolenylsulfenylamino) -3-carbamoylpropionyloxy] succinimide are obtained. 135 DEG-136 DEG C. (dec.).

3) 380 mg (1.00 mmol) of N- [D-2- (o-nitrol enylsulphenylamino) -3-carbamoylpropionyl] succinimide and 500 mg (1.07 mmol) of triethylamine salt of 6- [D-2] -amino-2- (p-hydroxyl enyl) acetamido] penicillanic acid is dissolved in a mixture of 4 ml of chloroform and ml of dimethylformamide. The solution was stirred at 0-5 ° C for 26 hours. After the reaction, the solution is evaporated at about 30 degrees Celsius under reduced pressure to remove the solvent. 10 ml of an aqueous 10% citric acid solution was added to the residue, and the mixture was extracted with a mixture of 20 ml of ethyl acetate and 10 ml of tetrahydrolurane. The extract was washed with water, dried and evaporated under reduced pressure at about 30 ° C to remove the solvent. 800 mg of 6- [D-2- (N-trifluoromethylsullenylamino) -3-carbamoylpropiramido) -2-p-hydroxylenylacetamido] penicillanic acid are obtained in the crude product. This crude product was dissolved in a mixture of 2 ml of tetrahydrolurane and 8 ml of ethanol. To this solution was added thiobenzamide (410 mL, 3 mmol) and the solution was stirred at room temperature for 15 minutes. The reaction mixture was evaporated under reduced pressure at about 30 ^o C. The residue is treated with 20 ml tetrahydroluranu and the bright yellow precipitate was isolated liltrací. The precipitate is dissolved in 30 ml of water, the insoluble matter is removed by filtration, and the filtrate is then freeze-dried. The colorless amorphous powder obtained is suspended in 3 ml of ethanol. 100 mg (1.0 mmol) of triethylamine and 180 mg (1.0 mmol) of potassium salt of α-ethyl capro are added for the first time. 20 ml of ether are then added and the colorless precipitate is collected by filtration. 370 mg of potassium salt of 6- [D-2- (D-2-amino-3-carbonylpropionamido) -2-p-hydroxylenylacetamido] penicillanic acid are obtained.

213 DEG-215 DEG C. (decomposition).

Infrared absorption spectrum:

ⁱⁿ nuP ¹ . _0j 330 1 ₇ ^{60, 1660,} 1590 c ^m1 '.

Thin-layer chromatography:

R f = 0.30 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)). .

Example 8

1) 594 mg (1.5 mmol) of N- [D-2- (o-nitrolenylsullenylamino) -3-N-methylcarbamoylpropionyloxy] succinimide and 900 mg (2.0 mmol) of the triethylamine salt of D-aminobenzylpenicillin are dissolved in a mixture of 25 ml of chloroform and 10 ml of dimethylformamide, the solution was stirred at 0-5 ° C for 17 hours. After reaction, the solution was worked up as described in Example 1-3. 870 mg of 6- [D-2- (DL-2- (o-nitro-phenylsulphenylamino) -3-N-methylcarbamoylpropirnamido) -2-0-enylacetamido] penicillanic acid are obtained in the form of yellow crystals. 135 DEG-140 DEG C. (decomposition).

Infrared absorption spectrum:

^at 3 ° ¹ : 3280 ^, 1780, 1730 ^, 1635 cm ^-1 .

2) 860 mg (1,35 µmol) of 6- [Da- (DL-2- / o-mirolenylsulphenylamino) -3-N-methylcarbamrylpropionamido) -2-1-enylacetamido] penicillanic acid and 547 mg (4 0.5 mmol) of thiobenzamide was dissolved in. mixture of 50 ml of methanol and 10 ml of tetrahydrolurane. The solution is stirred at normal temperature for 15 minutes. The reaction solution is then worked up in the same manner as in the example

1—4). 580 mg of 6- [D-2- (DL-2-amino-3-N-methylcarbamoylpropionamido) -2-0-phenylacetamide]] penicillanic acid are obtained in the form of a colorless powder.

Mp 193-195 ° C (dec.).

Infrared absorption spectrum:

.nu.max: 3250, 1760, 1645, 1590 cm @ -1.

Thin-layer chromatography:

R f = 0.45 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Example 9

1) 1.19 g (3 mmol) of N - [D-2- (1-mtrrlenylsullenylamino) -3-N-methyl] carbamrylprrpionylo-4] y] succinimide and 1.35 g (3 mmol) of the triethylamine salt of D-aminobenzylpenicillin are treated with Dissolve in a mixture of 25 ml of chloroform and 15 ml of dimethylformamide. The solution was stirred at 0-5 ° C for 15 hours. After completion of the reaction, the solution was treated in the same manner as in the example

1—3). There was obtained 1.81 g of 6- [D / 2 / (D-2- (o-nitrr) phenylsulphenyl-nitrile-3-N-methylcarbamoyl) propionamido) -2-lenylacetamido] penicyl202564 new as yellow needles. 145-147 ° C (dec.).

Infrared absorption spectrum:

nujol max.

^: 3250, 1780, 1725, 1630 cm ¹ .

2) 1.80 g (2.86 mmol) of 6- [D-2- (D-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionamido) -2-phenylacetylamido] penicillanic acid; and 1) 23 g (8.98 mmol) of thiobenzamide are dissolved in a mixture of 30 ml of methanol and 5 ml of tetrahydrofuran. The solution was stirred at normal temperature for 20 minutes. The reaction mixture was worked up as in Example 1-4. There were obtained 1.23 g of 6- [D-2- (D 1 - (1 R) - (R) - R (R) - (R) - (R, R) carbamoylpropionamido) -2-phenylacetamido]. m.p. 193 DEG-195 DEG C. (decomposition).

Infrared absorption spectrum:

in тал ¹ : 3275, 1760, 1650 cm ⁴ .

Thin layer chromatography:

Rf = 0.4-5 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)).

Infrared absorption spectrum of potassium salt:

nujol ⁱⁿ max.

: 3290, 1765, 1650, 1600 cm ⁴ .

Example 10

1) 1.60 g (4 mmol) of N- [L-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionyloxysuccinimide and 1.80 g of triethylamine salt of D-1a-aminobenzylpenicillin are dissolved in 25 ml. dimethylformamide. The solution was stirred at 0-5 ° C for 16 hours. After the reaction, the solution was worked up as in Example 1-3). 2.45 g of .alpha. (D, N- (4-Zo-nitrophenylsulphenylamino) -3-N-methylcarbamoylpropionamido) -2-phenylacetamido] pentanilic acid are obtained in the form of yellow needles. Melting point 144-146 ° C (decomposition).

Infrared absorption spectrum:

ⁱⁿ maX: 3250, 1770, 1730, 1635 cm ⁴ .

2] 2.40 g (3.81 mmol) of 6- [D-2- (L-2-O'-nitrophenylsulfenylamino-2-N-methylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid and 1.80 g (13.14 mmol) of thiobenzamide was dissolved in a mixture of 30 ml of methanol and 25 ml of tetrahydrofuran, and the solution was stirred at room temperature for 30 minutes, after which the reaction solution was worked up as described in Examples 1-4. 1.51 g of 6- [D-2- (L-2-amino-3-N-methylcarbamoylpropionamido) -2] acid are obtained.

-phenylacetamido] penicilane in the form of a colorless powder.

Mp 191-193 ° C (dec.).

Infrared absorption spectrum:

h: 3270, 1760, 1650 cm ⁴ .

Thin layer chromatography:

Rf = 0.45 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)).

Infrared absorption spectrum of potassium salt:

^v 2 ¹ : 3300, 1765 ^, 1650 ^, 1600 cm ⁴ , example 11

1) 1.6 g (10 mmol) of DL-2-amino-3 N - ^ - ^ e h ⁱ ylkarbamoylpiOpi ^ C ^ N ^ ew (i.e. DL-N-ethylasparagine) 2. g of nitrophenylsulphenyl chloride, 20 ml of water, 8 ml of tetrahydrofuran and 1.9 g of potassium carbonate are reacted as described in Example 1-1. 1.7 g of DL-2- (c-nitrophenylsulfanylmethyl) -butylpropionic acid are obtained.

178 DEG-180 DEG C. (decomposition).

2) 850 mg (2.7 mmol) of DL-2- (o-tert-enylamino) N-ethylcarbamoylpropionic acid, 577 mg of dicyclchexylcarbodiimide, 322 mg of N-hydrosuccinimide and 20 ml of tetrahydrofuran are reacted as described in Example 1 —2). 850 mg of [[E] -2- (c-nitrophenylsulfonylamino) -3-N-ethylcarbamoylpiperidinium-yuccinimide] are obtained in the form of a yellow caramel.

3) 820 mg (2 mmol) of N 1 [DL 12 - (o-nitro-phenylsulfenylamino) -3-N-ethylcarbamoyl-propynyl] succinimide and 900 mg (2 mole) of the lithium lithium salt of D-α-aminobenzyliline are dissolved in 15 ml of dimethyl. The solution was stirred at 0 to 5 degrees Celsius for 16 hours. After the reaction, the solution was worked up as in the example

1—3). There was obtained 1.19 g of 6- [D-2- (DL7-Dimethylphenylsulfrylamyl) -3-ethylcarbamoylpropionamide] -4-phenylacetamido] penicillanic acid as a caramel.

Infrared absorption spectrum:

ⁱⁿ 22. ¹ : 3270 ^, 1770 ^, 1720 ^, 1640 cm ⁴ .

4) 1.15 g. (1.78 mmol) of 6- [D-2- (DL12- (c-nitrophenylsulfonylamino) -3-N-ethylcarbamoylpropylpiperamide) -2-phenylacetamide] -pentilic acid and 822 mg (6 mmol) of thiobenzamide were dissolved in a mixture of 20 ml of methanol The solution was stirred at room temperature for 20 minutes and the reaction solution was then treated as in Example 1-4. 600 mg of 6- [D-2- (DL-2-amino-3-N-ethylcarbamoylpropionamido) -2-phenyl-acetamido] penicillanic acid are obtained.

Melting point 190-193 ° C (decomposition).

Infrared absorption spectrum:

¹ at 3270, 1760, 1650 cm ^1st

Thin layer chromatography:

R _f = 0.475 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)].

Infrared absorption spectrum of potassium salt:

May ^1: 3280, 1765, 1650, 1600 cm ¹ .

Example 12

1) 1.75 g (10 mmol) of DL-2-amino-3-N-propylcarbamoylpropionic acid (i.e. DL-N-n-propylasparagine), 2.2 g of o-nitrophenylsulfenyl chloride, 20 ml of water, 10 ml of tetrahydrofuran and 2.2 g of potassium carbonate were reacted in the same manner as described in Example 1-1. 2.2 g of DL-2- (-nitrophenylsulfenylamino) -3-N-n-propylcarbamoylpropionic acid are obtained. Melting point 200-205 ° C [decomposition].

2) 1.9 g (6 mmol) of DL-2- (o-nitrophenylsulfenylamino) -3-N-propylcarbamoylpropionic acid, 1.3 g of dicyclohexylcarbodiimide, 700 mg of N-hydroxysuccinimide and 20 ml of tetrahydrofuran are reacted as described above. in Example 1-2). 2.4 g of N- [DL-2- (o-nitrophenylsulfenylamino) -3-N-n-propylcarbamoylpropionyloxy] succinimide are obtained in the form of a yellow caramel.

3) 450 mg (1.06 mmol) of N- [DL-2- (o-nitrophenylsulfenylamino) -3-Nn-propylcarbamoylpropionyloxy] succinimide and 495 mg (1.1 mmol) of the triethylamine salt of D-aminobenzylpenicillin are dissolved in 15 ml dimethylformamide. The solution was stirred for 16 hours at 0-5 ° C. After the reaction, the solution was worked up as in Example 1-3). 230 mg of 6-

- [D-2- (o-Nitrophenylsulfenylamino) -3-N-n-propylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid as a caramel.

Infrared absorption spectrum:

ν _x ⁰¹ : 3280, 1770, 1725, 1645 cm ^-1 .

4) 210 m: g (0.31 mmol) of 6- [D-2-] acid

- (DL-2- (o-nitrophenylsulphenylamino) -3-N-n-propylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid and 137 mg (1.0 mmol) of thiobenzamide were dissolved in 10 ml of methanol. The solution was stirred at room temperature for 20 minutes. The reaction solution was worked up in the same manner as in Example 1-2. 75 mg of 6- [D-2- (DL-2-amino-3-N-n-propylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid are obtained. Melting point 175-178 ° C (decomposition).

Infrared absorption spectrum:

»S ^{1: 3250} ' ¹⁷⁶⁵ - ^{1650 cm} * ·

Thin layer chromatography:

_Rf = 0.55 [silica gel plate, solvent: n-butanol-acetic acid-water mixture (4: 1: 1)].

Example 1 a d 1 3

1) A solution of 13.5 g of DL-N- (o-nitrophenylsulfenyl) aspartic anhydride in 20 ml of tetrahydrofuran and 30 ml of methanol is heated at a temperature between 50-60 ° C for 30 minutes. The solvent was removed under reduced pressure. The residue is dissolved in 100 ml of tetrahydrofuran and 5.1 g of N-hydroxysuccinimide and 9.1 g of dicyclohexylcarbodiimide are added to the solution. The reaction mixture was then allowed to stand at room temperature for 17 hours. Precipitated urea was removed by filtration and the filtrate was evaporated under reduced pressure to remove the solvent. It is obtained

14.2 g of N- [DL-3- (o-nitrophenylsulfenylamino) -3-methoxycarbonylpropionoxy] succinimide.

M.p. 158-160 ° C (dec.).

2) A solution of 2.4 g of N- [DL-3- (o-nitrophenylsulfenylamino) -3-methoxycarbonylpropionyloxy] succinimide and 1.5 g of n-butylamine in 25 ml of tetrahydrofuran is allowed to stand at room temperature for 2 days. The solution was evaporated under reduced pressure to remove the solvent. 1.8 g of DL-2- (o-nitrophenylsulphenylamino) -3-N-n-butylcarbamoylproponic acid methyl ester are obtained.

Melting point 104-105 ° C.

3) 2 ml of hydrazine hydrate are added to a solution of 1.7 g of DL-2- (o-nitrophenylsulfenylamino) -3-N-n-butylcarbamoylpropionic acid methyl ester in 30 ml of methanol. After 1 hour the yellow crystals were collected. It is obtained

1.2 g of DL-2- (o-nitrophenylsulfenylamino) -3-N-n-butylcarbamoylpropionyl hydrazide. Melting point 180-182 ° C.

4) To a solution of 888 mg of DL-2- (o-nitrophenylsulfenylamino) -3-Nn-butylcarbamoylpropionyl hydrazide in 15 ml of tetrahydrofuran containing 5 ml of 10% sulfuric acid and 5 ml of acetic acid is added in portions at a temperature of -5 to -10. ° C 345 mg sodium nitrite. After 20 minutes, 20 mL of cold water was added to the reaction mixture. The reaction mixture is extracted with 20 ml of ethyl acetate. The extract was dried and then evaporated under reduced pressure to remove the solvent. 700 mg of DL-2- (o-nitrophenylsulphenylamino) -B-N-n-butylcarbamoylpropionylazide are obtained in the form of a yellow caramel.

5) 700 mg (1.9 mol) of DL-2- (o-nitrophenyl (sulfenylamino) -3-N- n-butylcarbamoylpropionylazide and 900 mg (2 mmol) of the triethylamine salt of D-.alpha.-aminobbenylpening of Cil are dissolved in 15 ml. The solution was stirred at 0-5 ° C for 48 hours, after which the solution was worked up as in Example 1-1). 210 mg of 6- [D-2- (DL 4 / o-nitrophenylsulfenylamino) -3 (N (n-butylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid are obtained as a yellow caramel.

Infrared absorption spectrum:

h: 3300, 1780, 1725, 1640 cm ¹ .

6) 200 mg (0.3 mmol) of 6- [D-2- (DL-2-o-nitrophenylsulphenylamino) -3-Nn-butylcarbamoylpropionamido) -2-phenylacetamido-iodicillanic acid and 300 mg (2.19 mmol) of thiobenzamide Dissolve in a mixture of 10 ml of methanol and 1 ml of tetrahydrofuran.

is stirred at room temperature for 30 minutes. Thereafter, the reaction solution was treated as described in Example 1-4. 85 mg of 6- [D-2- (D-2-amino-3-N-butylcarbamoylpropionamyl) -2-phenylacetamide] -penicillanic acid are obtained in the form of a colorless powder.

Melting point 190-193 ° C (decomposition).

Infrared absorption spectrum:

v тя? ^1: 3375, 1885, 1645 cm ^-1 .

Thin layer chromatography:

R f = 0.51 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)).

Example 14

(1) 1,76 g (10 mmol) of DL-2-amino-3-N-β-hydroxyethylcarbamoylpropionic acid (ie DL / N ^, - / - hydroxydroxylasparagine), 2,2 g of o-nitrophenyl chloride, 20 ml of water, 10 ml of tetrahydrofuran and 2.2 g of potassium carbonate were reacted as described in Example 1-1. 2.1 g of DL-2- (o-nitroftnylsulftnylamino) / 3 N / ^/ J4 / ydIΌoyythyllkaгbamoylpropionové. Melting point 123-125 ° C.

2) 1.64 g (5 mmol) of DL-2- (o-nitrophenylsulphenylamino) -3-N- (3-hydroxyedioxy) ethylcarbamoylpropionic acid, 1.13 g of dicyclohexylcarbodiimide, 633 mg of N-hydroxysuccinimide and 40 ml of tetrahydrofuran were reacted as described in Example 1-2. There was obtained 1.7 g of (3-N- (4-hydroxyethylcarbamoylpropionyloxy) succinimide) as a yellow, diamond-like substance.

3) 852 mg (2.0 mmol) of N- [DL-2- (o-nitrophenylsultenyaammo) -3-N (-1H-3-hydroxyethylcarbamoylpropionyloxy) succinimide 675 mg (1.5 mmol) of the triethylamine salt of D-benzylpe] - (?) The solution was dissolved in 30 ml of chloroform and stirred at 0-5 ° C for 16 hours, after which the solution was worked up as described in Example 1-3 to give 550 mg of 6- [6-] acid. D-2- (o-nitrophenylsulfenylamino) penicillanic acid as a yellow caramel.

Infrared absorption spectrum:

in me :? ^1: 3270, 1770, 1720, 1650 cm ^-1 .

4) 500 mg (0.76 mmol) of 6- [D-2- (DL-2- (o-nitrophenylsulfanylmino) -3-hydroxy-3-hydroxymethylcarbanoylpropionanido) -2-phenylacetyl] idojepenilic acid; 390 mg (2.8 mmol) of thiobenzamide are dissolved in a mixture of 10 ml of methanol and 2 ml of tetrahydrofuran. The solution was stirred at room temperature for 20 minutes. The reaction solution was then treated as described in the example

1—4). 280 mg of 6- [D-2- (DL / 2-anino: -3- [1,1-N- (3-ydyoxyethylcabamoylpro) pionamido) (2) phenylacttanide] penicillanic acid are obtained in the form of a colorless powder.

Melting point: 177-180 ° C (dec.).

Infrared absorption spectrum:

h: 3260, 1760, 1655 cm ¹ .

IHdX.

Thin layer chromatography:

Rf = 0.49 (silica gel plate, solvent: n-butane-acetic acid-water 4: 1: 1)].

Example 15

1) 1.61 g (10 mmol) of DL-2-amino-O (3-N-diethylcarbonyl-propionic acid (i.e. DL-N ^, -dinetolylsparagine), 2.1 g of c-nitrophenylsulfenyl chloride, 20 ml of water, 15 ml of tetrahydrofuran and 2.1 g of potassium carbonate were reacted as described in Example 1-1. 1.9 g of DL 4 - (c-nitrophenylsulfanylincin) -3-N-dintetylcarbbanoylproponic acid are obtained.

M.p. 144-146 ° C.

(2) 1.39 g (4.3 mmol) of DL-2- (o-nitrophenylsulfenylamino) -3-N-dimethylcarbamoylpiperidine, 9.36 mg of dicyclicexylcarbamide, 518 mg of N-hydroxysuccininide and 40 ml of tetrahydrofuran are reacted as described in Example 1-2. 1.6 g of N- [DL-2- (c-nitrophenylsulfonylamino) -3-N (diethylcarbamylpiperidinyl) succinide are obtained.

126 DEG-128 DEG C. (decomposition).

3) 820 mg (2 mmol) of N- [DL-2 ((o-nitroft (nylsulfenyl lamino) -3-N-dimethylcarbamoylpropionyloxy) succinimide) and 900 mg (2 mmol) of triethylamine salt of D-benzylpenicillin are dissolved in a mixture of 15 ml of chloroform and 13 ml of dimethylformamide, and the solution was stirred at 0-5 ° C for 16 hours, after which the solution was worked up as in Example 1-1 to give 1.22 g of 6- [D-2- (DL-2). - (o-Nitrophenylsulfenylamino) -3-N-dimethylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid as a yellow caramel.

Infrared absorption spectrum:

h ¹ : 3200, 1780, 1730, 1630 cm ¹ .

4) 1.21 g (1.88 mmol) of 6- [D-2- (DL-2- (o-nitrophenylsulfenylamino) -3-N-dimethylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid and 1.03 g (7.5 mmol) of thiobenzamide are dissolved in a mixture of 30 ml of methanol and 10 ml of tetrahydrofuran. The solution was stirred at normal temperature for 20 minutes. Thereafter, the reaction solution was treated as described in Example 1-4. 660 mg of 6-f-2- (DL-2-amino-3-N-dimethylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid are obtained.

Mp 192-194 ° C (dec.).

Infrared absorption spectrum:

h 3240, 1770, 1630 cm ¹ .

Thin layer chromatography:

R ₍ = 0.451 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio))].

Infrared absorption spectrum of potassium salt:

in max * ^{1: 32401 17601 16401 1580 cra1} '

Example 16

(1) 720 mg (2 mmol) of DL-2-benzyloxycarbonylamino-3-benzyloxycarbonylpropionic acid and 210 mg (2 mmol) of triethylamine are dissolved in 7 ml of dichloromethane and a solution of 270 mg (2 mmol) of isobutylchlorocarbonate in 2 ml of dichloromethane is added. temperature to -20 to -15 ° C. 10 minutes after the addition, a solution of 740 mg (2 mmol) of triethylamine salt of D-.alpha.-aminobenzylpenicillin in 4 ml of dimethylformamide was added dropwise. The reaction mixture is stirred at -15 to -10 ° C for 30 minutes and then at -10 to 0 ° C for 30 minutes. After the reaction, the reaction mixture is concentrated under reduced pressure at about 30 ° C to remove the solvent. 5 ml of 5% aqueous citric acid solution and 10 ml of ethyl acetate are added to the obtained residue and the organic solvent layer is separated. The organic solvent layer is washed with water, dried and then mixed with 20 ml of triethylamine (2 mmol). The mixture thus obtained was evaporated under reduced pressure at a temperature of about 30 ° C to remove the solvent. 970 mg of triethylamine salt of 6- [D-2- (DL-2-benzyloxycarbonylamino-3-benzyloxycarbonylpropionamido) -2-phenylacetamido] penicillanic acid are obtained in the form of colorless crystals.

Melting point 110-113 ° C (decomposition).

Infrared absorption spectrum:

in max ^{: 33001 17751 17351 16901 16101} 1520 cm ¹

(2) 200 mg (0.3 mmol) of triethylamine salt of 6- [D-2- (DL-2-benzyloxycarbonylamino-3-benzyloxycarbonylpropionamido) -2-phenylacetamido] penicillanic acid are dissolved in 6 ml of ethanol and 300 ml are added to the solution. mg of 30% palladium on barium carbonate. The solution was shaken at room temperature in a hydrogen atmosphere for 4 hours at atmospheric pressure. The insoluble material after the reaction was removed by filtration and the filtrate was evaporated under reduced pressure at about 30 ° C to remove the solvent. To the residue is added 5 ml of ether. The colorless precipitate thus obtained is isolated by filtration, washed with chloroform and dried. 60 mg of triethylamine salt of 6- [D-2- (DL-2-amino-3-hydroxycarbonylpropionamido) -2-phenylacetamido] penilcianoic acid are obtained.

Melting point 189-191 ° C (decomposition).

Infrared absorption spectrum:

v nuM. _{3300 1750 1665 1595 1540} Шал.

cm ¹ .

Thin layer chromatography:

_Rf = 0.36 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Example 17

1) 9.2 g (50 mmol) of DL-2-amino-3-methoxycarbonylpropionic acid hydrochloride (i.e. DL-aspartic acid O-N-methyl ester hydrochloride), 10.5 g of o-nitrophenylsulfenyl chloride, 50 ml of water, 70 ml of tetrahydrofuran and 24.3 g of potassium carbonate were reacted in the same manner as in Example 1-1. 9.8 g of DL-2- (o-nitrophenylsulfenylamino) -3-methoxycarbonylproponic acid are obtained.

Melting point 108-109 ° C.

2) 9 g (30 mmol) of DL-2- [o-nitrophenylsulfenylamino) -3-methoxycarbonylpropionic acid, 6.7 g of dicyclohexylcarbodiimide, 4 g of N-hydroxysuccinimide and 80 ml of te202564 trahydrofuran are reacted as in Example 1. —2). 11 g of N- [DL-2- (o-nitrophenylsulfenylamino) -3-methoxycarbonylpropionyloxy] succinimide are obtained. Melting point 125-127 ° C.

3) 794 mg (2 mmol) of N- [DL-2- (o-nitrophenylsulfenylamino) -3-methoxycarbonylpropioinyloxy] succinimide and 900 mg (3 mmol) of the triethylamine salt of N 2 - (1-nitrophenylsulfenylamino) -3-methoxycarbonylpropioinyloxy-succinimide. The piperidine is dissolved in a mixture of 25 ml of chloroform and 4 ml of dimethylformamide. The solution was stirred at 0-5 ° C for 15 hours. After the reaction, the solution was treated as described in Example 1-3). 1.2 g of 6- [D-2- (DL-2- (o -trophenylsulfenylamino) -3-methoxycarbonylpropionamido) -2-phenylacetamido] penicillanic acid are obtained in the form of a yellow caramel.

Infrared absorption spectrum:

in liquid max.

: 3300, 1780, 1725, 1650 cm ¹ .

4) 1.2 g (19 mmol) of 6- [D-2- (DL-2- (o -trophenylsulfenylamino) -3-methoxycarbonylpropionamido) -2-phenylacetamido] penicillanic acid and 800 mg (5.84 mm.ol) thiobenzamide was dissolved in a mixture of 20 ml of methanol and 3 ml of tetrahydrofuran. The solution was stirred at room temperature for 45 minutes. Thereafter, the reaction solution was treated as in Example 1-4). 1.51 g of 5- [D-2- (DL-2-amino-3-methoxycarbonylpropioinamido) -4-phenylacetamido] penicillanic acid are obtained in the form of a colorless powder, m.p. 185 DEG-188 DEG C. (dec.

Infrared absorption spectrum:

in T ³ * ¹ : 3250, 1760, 1730, 1660 cm ^-1 .

Thin layer chromatography:

R _f = 0.453 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)).

Infrared absorption spectrum of potassium salt:

v ^ ° ^l: 3280, 1760, 1730, 1650, 1595 cm ^4th

In the same manner as described above, the following compound was prepared:

6- [D-2- (D-2-Amino-3-methoxycarbonylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid. Melting point 170-173 ° C (decomposition).

Infrared absorption spectrum:

in hats: 3330, 1760, 1720, 1650, 1600 cm ⁴ .

Thin layer chromatography:

Rj = 0.42 [silica gel plate, solvent 26 n-butanol-acetic acid-water (4: 1: 1 ratio)].

Example 18

(1) 18,4 · g of D-2-amino-3-methoxycarbonylproponic acid hydrochloride (ie D-aspartic acid O-methyl ester hydrochloride), 2 g of o-nitrophenylsulphenyl chloride, 80 ml of water, 80 ml of tetrahydrofuran and 26 g of potassium carbonate are reacted as in Example 1 - 1). 18 g of D-2- (o-nitrophthylsulphenyl / amino) -3-methoxycarbonylpropionic acid are obtained. Melting point 86-87 ° C.

2) 3 g (10 mmol) of D-2- (o-nitrophenylsulfanylamino) -3-methoxycarbonylpropionic acid, 2.3 g of dicyclohexylcarbodiimide, 1.35 g of N-hydro (xysuccinimide and 30 ml of tetrahydrofuran) are introduced into reaction as in Example 1-2). 3.5 g of N- [D-2- (o-nitrophenylsulfenylamino) -3-methoxycarbonylpropionyloxy] succinimide are obtained. Melting point 133 ° C [decomposition],

3) 794 mg (2 mm &lt; 3 &gt;) of N- [D-2- (o-mtrophthylsulphonylamino) -3-methoxycarbonylpropionyloxysuccinimide and 900 mg (2 mmol) of triethylamine salt of D-.alpha.-aminobenzyipenicillin are dissolved in 25 ml of chloroform. The solution was stirred for 16 hours at 0-5 ° C. After the reaction, the solution was treated as in Example 1-3). There was obtained 1.15 g of 6- [D / 2- (D-2- (o-maphthophenylsulphenylamino) -3-methoxycarbonylpropenamido) -2-phenylacetyl] penicillanic acid as a caramel solid.

Infrared absorption spectrum:

Й ¹ : 3270, 1775, 1720, 1640 cm ⁴ .

4) 1.10 g (1.74 mmol) of 6- [D-2- (D-2-Z-nitrophenylsulphenylamino) -3-methoxycarbonylpropionamido) -2-phenylacetamidephenicillanic acid and 835 mg (6.1 mmol) thiobenzamide was dissolved in a mixture of 30 ml of methanol and 5 ml of tetrahydrofuran. The solution is stirred for 45 minutes at normal temperature. The reaction mixture is then worked up as in Example 1-4). 680 mg of 6- [D-2-amino-3-methoxycarbonylpropionamido) -2-phenylacetamido) -pentilic acid are obtained.

M.p. 185-188 ° C (dec.).

Infrared absorption spectrum:

h ^: 3250 ^, 1765, 1725 festerL 1655 cm * ¹ .

Thin layer chromatography:

R _f = 0.453 (silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)).

Example 19

1) 720 mg (2 mmol) of DL-2-benzyloxycarbonylamino-3-carboxypropionic acid benzyl ester and 210 mg (2.1 mmol) of triethylamine are dissolved in 7 ml of dichloromethane and a solution of 270 mg (2 mmol) of isobutyl chloroformate is added. -20 to -15 ° C. 10 minutes after this addition, a solution of 990 mg (2.2 mmol) of the triethylamine salt is added dropwise thereto. The reaction mixture is then treated as in Example 16-1). 1.30 g of triethylamine salt of 6- [D-2- (DL-3-benzyloxycarbonylpropionamido) -2-phenylacetamido] penicil- iate are obtained. new in the form of a colorless powder. 115 DEG-119 DEG C. (decomposition).

Infrared absorption spectrum:

ⁱⁿ max ¹ ' ^: 3300, 1775, 1730, 1660, 1610,

1500 cm ⁴ .

2) • 660 mg (0.83 mmol) of triethylamine salt of 6- [D-2- (DL-3-benzyloxycarbonylamino-3-benzyloxycarbonylpropionamido) -2-phenylacetamido] -penicillanic acid are dissolved in 20 ml of ethanol and dissolved in a solution. 500 mg of 30% palladium on barium carbonate are added. The solution was shaken at room temperature under a hydrogen atmosphere at atmospheric pressure for 2 hours. Thereafter, the reaction mixture was worked up as in Example 16-2). 220 mg of triethylamine salt of 6- [D-2- (DL-3-amino-3-hydroxycarbonylpropionamido) -2-phenylacetamido] penicillanic acid are obtained in the form of an off-white powder. Mp 185-188 ° C (dec.). Infrared absorption spectrum:

ⁱⁿ max ¹ : 32 ^, 50> 1770 ^- 1650, 1600, 1530 cm ⁴ .

Thin layer chromatography:

_Rf = 0.28 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

Example 20

1) 1.6 g (10 mmol) DL-3-amino-3-N-ethylcarbamoylpropionic acid (i.e. DL-N'-ethylisoasparagine), 2.2 g o-nitrophenylsulfenyl chloride, 20 g water, 20 ml tetrahydrofuran and 2 g 3 g of potassium carbonate were reacted as in Example 1 - 1). 2.0 g of DL-3- (o-nitrophenylsulphenylamino) -3-N-ethylcarbamoylpropionic acid are obtained.

Melting point 165-167 ° C (decomposition).

2) 1.05 g (3.35 mmol) of DL-3- (o-nitrophenylsulfenylamino) -3-N-ethylcarbamoylpropionic acid, 700 mg of dicyclohexylcarbodine, 391 mg of N-hydroxysuccinimide and 40 ml of tetrahydrofuran are reacted as in Example 1-2). It is obtained

1.2 g of N- [DL-3- (o-nitrophenylsulfenylamino) -3-N-ethylcarbamoyl] -pionyloxy] succinimide.

M.p. 157-158 ° C.

3) 800 mg (2 mmol) of N [[DL-3- (o-n-triphenylsulfenylamino) -3-N-ethylcarbamoylpropionyloxy] -succinimide and 900. Dissolve mg (2 mmol) of triethylamine for the first time in D-α-aminobenzylpenicillin salts in 15 ml of dimethylformamide. The solution is stirred for 16 hours at 0 to > 5 ° C. After the reaction, the reaction solution is treated as described in Example 1-3). 1.09 g of 6- [D-2- (DL--3- (o-nitrophenylsulphenyl) lamino] -3-N-ethylcarbamoylpropionamide) -2-phenylacetamide-enenillanilic acid are obtained in the form of a caramel.

Infrared absorption spectrum:

in max ^1: 3260-1775, 1725, 1620 cm ⁴ .

4) 1.05 g (16 mmol) of 6- (D-2-) acid

- enylsulfenylamino / -3-N-ethylcarbamrylprrpirnamol-2 '- 2-phenylacetamido -penicilane and 781 mg [5.7. • mmol) of thiobenzamide is dissolved in a mixture of 30 ml of methanol and 5 ml of tetrahydrofuran. The solution was stirred at room temperature for 45 minutes. Thereafter, the reaction mixture was worked up as described in Example 1-4. 730 mg of 6-D-2 - [(DL-3-amino-3-N-ethylcarbamoylpropionamido) -2-phenylacetamido] -penicillanic acid are obtained.

Melting point 200-203 (decomposition).

Infrared absorption spectrum:

ν ° ^1: 3890 ^, 1765 ^, 1655 cm ⁴ .

Thin layer chromatography:

R _f = 0.475 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1 ratio)].

Example 1 a d 2 1

1) 1.75 g (10 mmol) of DL-3-amino-3-Nn-propylcarbamoylpropionic acid (i.e. DF-n-propylasparagine), 2.2 g of o-nitrophenylsulfenyl chloride, 20 ml of water, 20 ml of tetrahydrofuran and 2 3 g of potassium carbonate are reacted as in Example 1 - 1). 2.8 g of DL-3- [o-nitrenylsulfenylamino) -3-N-n-propylcarbamoylpropionic acid are obtained.

148 DEG-150 DEG C. (decomposition).

2) 981 mg (3 mmol) of DL-3- (o-nitrophenylsulphenylamino) -3-Nn-propylcarbamrylpropionic acid, 618 mg of dicyclohexylcarbodiimide, 345 mg of N-hydroxysuccinimide and 40 ml of tetrahydrofuran are reacted as described in Example 1-2). 1.1 g of N- (DL-3- (o-nitrophenylsulfenylamino) -N-n-n-propylcarbamoylpropionyloxy] succinimide are obtained in the form of a yellow caramel.

3] 848 mg (2 mmol) of N- [DL-3- (o-nitrophenylsulfenylamino) -3-Nn-propylcarbamoylpropionyloxy] succinimide and 900 mg (2 mmol) of the triethylamine salt of D-benzylpenicillin are dissolved in 20 ml of dimethylformamide. for 16 hours at 0-5 [deg.] C. After the reaction, the solution is worked up as described in the example

1—3). 800 mg of 5- [D-2- (DL-37-nitrophenylsulfenylamino) -3-N-n-propylcarbamoylpropionamido] penicillanic acid are obtained in the form of a yellow caramel.

Infrared absorption spectrum:

v. 3275, 1775, 1725, 1635 cm -1.

4) 780 mg (1.17 mmol) of 6- [D-2- (D ^ - ^ - ^ - (^ - nk: rofenylsulfeny] and ^ m ^ ^ r ^ o _/ ^/ - ^ 3-Nn -propylcarbamoylpropionamido] percilanic acid and 561 mg (4.1 mmol) of thiobenzamide are dissolved in a mixture of 15 ml of methanol and 2 ml of tetrahydrofuran.

The solution was stirred for 20 minutes at room temperature. Thereafter, the reaction mixture was worked up as described in Example 1-4. 390 mg of 6- [D-2- (DL-3-amino-3-Nn-propylcarbamoylpropionamido) -2-phenylacetamiido] penicillanic acid are obtained. Mp 181-184 ° ^C (dec):

Infrared absorption spectrum:

nujol max.

: 3280, 1770, 1660 cm-1,

Thin-layer chromatography:

Rf ₌ 0.55 [silica gel plate, solvent: n-butanol-acetic acid-water (4: 1: 1)].

3) 820 mg (2 mmol) of N- [DL-3- (o-nitrophenylsulfenylamino) -3-N-dimethylcarbamoylpropionyloxy] succinimide and 900 mg (2 mmol) of the triethylamine salt of Da-amino are dissolved in a mixture of 25 ml of chloroform. and 12 ml of dimethylformamide. The solution was stirred for 18 hours at 0 to 5 · ^Q C. After the reaction solution was treated in the same way SE- · as described in Example 1 -3). There was obtained 1.25 g of 6- [D-2- (DL-7-O-010-α-dimethylamino) -2-phenylacetamido] penicillanic acid as a yellow caramel.

Infrared absorption spectrum:

ⁱⁿ max ^: 3270, 1780, 1725, 1630 · cm ¹ .

4) 1.20 g (1.86 mmol) of 6- [D-2- (DL-3- (o-nitrophenylsulfenylamino) -3-N-dimethylcarbamcylpiperidine] -2-phenylacetamido] pentanilic acid and 800 mg (5.8 mmol) of thiobenzamide was dissolved in a mixture of 20 ml of methanol and 3 ml of tetrahydrofuran. The solution was stirred for 30 minutes at room temperature. The reaction solution is then worked up as described in Example 1-4). 810 mg of 6- [DL-3-amino-3-N-dimethylcarbamoylpiperamidamide] -2-phenylacetamido] penicillanic acid is obtained in the form of a colorless powder.

207 DEG-210 DEG C. (decomposition).

Infrared absorption spectrum:

^v , ^I ™. ^{J * I:} 3250, 1760, 1645 cm ^{@ 4} .

Thin layer chromatography:

Rf = 0.392 [silica gel plate, solvent: n-butane-acetic acid-water (4: 1: 1 ratio)].

Example 22

Infrared spectrum · potassium salts:

1) 1.6 g (10 mmol) of DL-3-amino-3-methyl-4-amino-3-amino-3-amino-3-amino-3-amino-3-amino-3-amino-3-amino-3-amino-3-amino g of c-nitrophenylsulfenyl chloride, 20 ml of water, 20 ml of tetrahydrofuran and 2.2 g of potassium carbonate were reacted as in Example 1-1). 1.9 g of DL-3- (c-nitphenylsulfenylamino) -3-N-n-dimethylcarbamnylpropionic acid are obtained.

M.p. 144-146 ° C.

2) 1.88 g (6 mmol) of DL-3- (o-nitrophenylsulfenylamino) -3-N-dimethylcarbamoylpropionic acid, 124 g of dicyclohexylcarbodiimide, 748 g of N-hydroxysuccinimide and 40 ml of tetrahydrofuran were reacted as in Example 1 —2). 900 mg of N- [DL-3- (o-terferylsulfenylamino) -3-N-dimethylcarbamoylpiperidine] succinimide are obtained.

Melting point · 171-172 ° C, nu jol max.

: 3260, 1760, 1640, 1595 cc.

Example · 23

2.99 g (10 millimoles) of D-2- (o-nitrophenylsulfenylamino) -3- (N-methylcarbamoyl) propionic acid and 1.11 g (11 millimoles) of triethylamine are dissolved in 30 ml of dimethylformamide and the acid azide is added. diphenylphosphoric acid (11 millimoles) at 0-5 ° C. The mixture was stirred at the same temperature for 20 minutes. 4.95 g of the triethylamine salt of D-.alpha.-amincbenzylpenicillin are added and the mixture is stirred at the same temperature for 3 hours and then at room temperature for 1 hour. After completion of the reaction, 150 ml of water and 303 mg (3 millimoles) of triethylamine were added to the reaction mixture, and the aqueous mixture was washed with 30 ml of ethyl acetate for 20.5%. The aqueous layer was then separated, adjusted to pH 3.5 with citric acid and extracted twice with a mixture. tetrahydrofuranethyl acetate (1: 1). The extract is washed with water, dried and then evaporated at a temperature not exceeding 40 ° C under reduced pressure to remove the solvent. 20 ml of ether-dichloromethane (1: 1) was added to the obtained residue, and the crystalline precipitate was collected by filtration and dried in vacuo to give 5.80 g of 6- [D-2- (D-2- / o-nitrophenylsulphenyl) acid. lamino [3-N-methylcarbamoylpropionamido] -2-phenylacetamido] penicilane in the form of yellow needles The physicochemical properties of this product are identical to those obtained in the example.

This product was treated as in Example 1-2 to give 6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid as colorless form. powder.

Melting point 198 to 201 ^and C (dec).

Example 1 a d 2 4

Dissolve 488 mg of triethylamine salt of D-2- (2-methoxycarbonyl-1-methylvinylamino) -3-N-methylcarbamoylpropionic acid in 10 ml of chloroform and add a solution of 274 mg of isobutyl chloroformate in 2 ml of chloroform at -20 to -15 ° C. Three minutes after the addition, 967 mg of 6- [D-2-amino-2- (p-hydroxyphenyl) acetamido] penicillanic acid phenacylate was added to the solution, followed by stirring at -5 to 0 degrees Celsius for 30 minutes at at 0 DEG C. for 1 hour After reaction, the mixture is evaporated at a temperature not exceeding 40 degrees Celsius to remove the solvent to give 6- [D-2- (D-2- / 2-phenylacetate) phenacyl ester as a crude product. -methoxycarbonyl] 11-methylvinyl] penicillanic.

ml of tetrahydrofuran and 5 ml of 0.5% hydrochloric acid were added to the above crude product and the mixture was stirred for 1 hour. To the mixture was added 5 mL of ethyl acetate. Then; shake the mixture. The aqueous layer was separated from the mixture, adjusted to pH 10 with aqueous sodium bicarbonate solution while cooling, and extracted with a mixture of 10 mL of tetrahydrofuran and 5 mL of ethyl acetate. The extract was washed with water, dried and then evaporated at a temperature not exceeding 40 ° C to remove the solvent. There was thus obtained 6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamide] penicillanic acid phenacylate as a crude product. To a crude product of 6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid phenacylate was added a solution of 264 mg of thiophenol in 6 ml of dimethylformamide and allowed to stand at temperature at 16 ° C for 40 minutes. To the reaction solution was added 15 ml of acetone. The resulting pale yellow precipitate was collected by filtration and dried. 537 mg of 6- [D-2- [2- [2- (10,10-O-1,4-dimethyl-propionamido) -2-p-hydroxyphenylacetamido] -2-p-hydroxyphenylacetamido] penicillanic acid sodium salt is obtained. The physicochemical properties of this product are identical to the compound obtained in Example 2-4].

Example 1 a d 2 5

957 mg of D-2- (o-nitrophenylsulfenylamino) -3- (N-methylcarbamoyl) -propionic acid 2,4,5-trichlorophenyl ester and 1,048 g (2 millimoles) of phthalimidomethyl ethyl ester of 6- [D-2-amino-2- (p- The hydroxyphenyl i) acetamido] penicillanic acid is dissolved in 10 ml of dimethylformamide, stirred at 0-5 ° C for 18 hours, and 30 ml of water, 25 ml of tetrahydrofuran and 20 ml are added to the reaction solution. The organic layer of solvent was separated from the mixture and washed with 10 ml of 5% aqueous citric acid solution, 10 ml of aqueous sodium bicarbonate solution and 10 ml of saturated aqueous sodium chloride solution, then the organic layer was dried and evaporated at room temperature. To the residue is added ether and the precipitate is collected by filtration to give 1.017 g of 6- [D-2- (D-2-o-nitrophenylsulphenylamine) phthalimidomethyl ester of phthalimidomethyl ester. 3-N-methylcarbamoylpropi onamido) -2-hydroxyphenyl] acetic acid in the form of yellow needles.

105 DEG-107 DEG C. (decomposition).

Infrared absorption spectrum:

ⁱⁿ ^ ° ^{1: 3250, 1780,} 1725, 1640 ^cm-first

Thin layer chromatography:

R ₍₌ 0.72 (silica gel plate, solvent: ethyl acetate-tttIahydrofuIan- acetic acid (10: 10: 1).

1.63 g of 6- [D-2- (D-2-O-nitrophenylsulfenylamino-3-N-methylcarbamoyl-p-opionamido) -2-p-hydroxyphenylacetamido] penicillanic acid phthalimidomethyl ester and 500 mg of n-octyl-2-mercaptoic sodium salt The solution was dissolved in 10 ml of dimethylformamide. The solution was stirred at 0-5 ° C for 40 minutes. To the reaction mixture were added 20 ml of water and 10 ml of ethyl acetate, and the mixture was shaken. The aqueous layer was separated from the mixture, acidified with 5% aqueous citric acid solution and extracted with 30 ml of a 2: 1 mixture of tetrahydrofuran ethyl acetate. The extract was washed with saturated aqueous sodium chloride solution, dried. and then evaporating at a temperature not exceeding 40 ° C to remove the solvent. Ether was added to the residue at 202564 and the precipitate was collected by filtration. 1.0 g of 6- [D-2- (D-2- (o-nitrophenylsulfenylamino) -3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid is obtained in the form of yellow needles.

This product was treated in the same manner as described in Example 2-4 to give 6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicllanic acid. Mp 198-201 ° C (dec.).

Example 26

1) 285 mg of D-2-benzyloxycarbonylamino-3-carbamoylpropionic acid, 115 mg of N-hydroxysuccinimide and 101 mg of N-methylmorpholine are dissolved in 2 ml of dimethylformamide and 137 mg of isobutyl chloroformate are added dropwise with stirring. After five minutes, a solution of 6- [D-2-amino-2- (p-hydroxyphenyl) -acetamido] penicillanic acid benzyl ester in 1 mL of dimethylformamide was added to the mixture, followed by stirring at -5 to 5 ° C for 30 minutes and 1 hour. The reaction was quenched with saturated aqueous sodium chloride solution and extracted with 5 mL of tetrahydrofuran and 3 mL of ethyl acetate, and the extract was washed with dilute aqueous citric acid, aqueous sodium bicarbonate, saturated aqueous sodium chloride. The extract was dried and evaporated at a temperature not exceeding 40 ° C to remove the solvent, ether was added to the residue to give 670 mg of 6- [D-2-benzyloxycarbonylamino-2-carbamoylpropionamido) benzyl ester. 2-p-hydroxyphenylacetamido] penicilane as a colorless crystalline powder.

Melting point 100-102 ° C (dec.). Yield 95%.

Infrared absorption spectrum:

ⁱⁿ max ^{1: 3280} ' ^{17701 17401 16951 1645} cm ¹ .

Thin layer chromatography:

R _f = 0.45 (silica gel plate, solvent: ethyl acetate-tetrahydrofuran (2: 1)

2) 335 mg of 6- [D-2- (D-2-benzyloxycarbonylamino-3-carbamoylpropibamido) -2-p-hydroxyphenylacetamido] penicillanic acid benzyl ester and 300 mg of 30% palladium on carbon are suspended in 3 ml of methanol. The suspension is shaken for 1 hour under an atmosphere of hydrogen at atmospheric pressure. After the reaction, the catalyst is filtered off and the filtrate is evaporated at a temperature not exceeding 40 ° C to remove the solvent. 3 ml of tetrahydrofuran are added to the residue, the crystalline precipitate is collected by filtration and washed with tetrahydrofuran. 200 mg of 6- [D-2- (D-2-amino-3-carbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid are thus obtained.

Melting point 202-205 ° C (decomposition).

Example 27

1) 994 mg of 2,2,2-trichloroethyl ester of 6- [D-2-amino ^L 2- (p-hy droxyfenylacetamido] penicillanic acid was added to 20 ml of a dimethylformamide solution containing 556 mg of D-2-benzyloxycarbonylamino-3-N 412 mg of dicyclohexylcarbodiimide are added with stirring at -5 to 0 DEG C. and the mixture is stirred at the same temperature for three hours, after which the precipitate is filtered off, 50 ml of water are added to the filtrate and the aqueous mixture is extracted. The mixture was washed with 5% aqueous citric acid solution, aqueous saturated sodium bicarbonate solution, dried and evaporated to remove the solvent, ether was added to the residue, and the precipitate was filtered and washed with ether. It is obtained

1.39 g of 6- [D-2- (D-2-benzyloxycarbonylamino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid 2,2,2-trichloroethyl ester as a yellow crystalline powder.

Melting point 173-175 ° C (dec.).

Infrared absorption spectrum:

^h max ^{01: 33001 17751 17001 1650 cm l} ·

Thin layer chromatography:

R _f = 0.75 (silica gel plate, solvent: ethyl acetate-tetrahydrofuran 2: 1).

2) 758 mg of 6- [D-2- (D-2-benzyloxycarbonylamino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid 2,2,2-trichloroethyl ester and 650 mg of zinc dust are added to 30 ml of 90% formic acid. The mixture was stirred at 0 ° C for 40 minutes. After the reaction, water (50 ml) was added and the mixture was extracted with tetrahydrofuran (30 ml) and ethyl acetate (30 ml). The extract was washed with water. A solution of 500 mg of triethylamine in 15 ml of water is then added to the extract. After shaking the mixture of said extract and triethylamine solution, the aqueous layer is separated, the pH is adjusted under cooling with 5% citric acid, and again extracted with a mixture of 20 ml of tetrahydrofuran and 15 ml of ethyl acetate. The extract thus obtained was washed with water, dried and evaporated to remove the solvent. Ether was added to the residue to give 266 mg of 6- [D-2- (D-2-benzyloxycarbonylamino-2-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid.

Infrared absorption spectrum:

ⁱⁿ mi? ^1: 3280, 1770, 1720, 1640 cm ^-1 .

Thin layer chromatography:

^Rf plate of silica gel, solvent tetrahydrofuran-acetic acid {50: 3: 3).

3) O- [D-2- (D-2-Benzyloxycarbonylamino) -3-N-methylcarbamoylpropionamido) -2-hydroxyphenylacetamido] penicillanic acid was treated as described in Example 27-2) to treat to give O- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamide] penicillanic acid.

Melting point 198 - 201 ° C (decomposition).

it

Pí ф

ω Си О

I >> <

д .Ц ф%

Ξ3 тз Д е

cd cd Ó д д

Е g я

3 °

4- »

В ό ό

Д Д. □ са -5 я -д я g я · = я · ±

ЕйЕсЕ £ я.йЕйЕ яшА · г * 4 * ^Г Т 'Гг СИ £> Ч Р Р и *' ¹

Д

СЛ 4- »ф в дц Д>. Д о tť, Си

Рч о сл

Во о £ д .д

Д CVd Д-д g я Е яВ 'Т * гт <' - rrt j> ^ d о сл ф ^с гД д

сл сл .Р

В cd

Cd cd Д cd сл х; Л i όχ ⁽ сл

X X

О О X Д Д о 4-> 4— · ф ф

Е 6

it it it it it it it it Pi Pi Pi Pi Pi Pi Pi 'Pi O O O O O O O O O ČM ω см У O cq O eq O cq O O Cm cm X X X E X X X X O O O ω Q O o ω

ZZZZ z Z ^Z Z

X X OF X X X X X O O Q O O O O O O O O O O O O O ω ω AT O O ω O o

it Pi it Pi it Pi it Pi it Pi it Pi it Pi tO 10 n »Pi Pi Pi Pi Pi O O O O O O O O O O O O cq o Cm at cm O cm O CM O CM O Ol O O O - X X X X X X OF X X I £ O O O ω ω O O ^{ο ω X} eq X eq E cm X eq E CM X ČM X CM E T? 'M oj XXX 4Γ zzz ^p OF OF OF of of of OF X X X X X X OF X X X X O O O O ω O ω O O O O O O O O O O O o o o o O ω O O ω O ω o o o u

<N tO

Pi

Ό T3

CD До S Sw> £ £ £ Pi

it Pi it Pi it Pi it Pi it Pi O O O O O O O O O ω cm Cm cm cm cm E E X X X O O ω ω ω r— ' / —X / —5 _t -------- p г — 4 N N N N N X X X X X of of OF OF OF '-' '-' X_______t X______t X X X X X ω O O O O O O O O O at ω ω O O

X X X X X X X X X 5 O O O O O r4 ¢ 4 VI m

X

X O

XXX

CM>

& & O

it Pi it it Pi Pi it Pi it Pi O O O O O O ω o O O ČM Cm CM ČM eq X X X X X O o o O O / -> / —ч N N N N N X X X X X OF Z Z OF OF '--------AND *________F ON ⁴ - ' X X X X X O ω o O O O o o O O O o o at ω

it Pi it Pi it Pi it Pi it Pi O O O O O O ω O O O cm eq cm eq eq X X E E E O O O O ω > r ~ \ F- t ----- X N tM N N N X X X X X Z _z of OF OF of X X X X X O O O O at O O O O O O O O O at

to c X có tó ~ Π °° η Huo '? О cm eq 4? xx X - N ^N S йшдг | ee §

X ω o ω

I ee§uu E oo E QO _O ¹¹ O

XX §X

X ^XX

X ^f XX

X

What about 00 σ>

ТГ 1П r- rH co t>. co cd rH τ-H r-rH o τ — i OJ CO

Ή rH rH rH

it

P4

what the

P4 O ω

Ю tO

O cd

СЧ

Z Z <N

Z Z z z z z

O CD

WHAT

From cd it

O СЭ what of o bo

O ω

WHAT

X

O to t o O O O O what what Z X u ω φ

CD-4 O N>

>> and • гИ t Ф> o

About what

r4 honors o ss> øx> Q PL |

Z o what π o o o

AND

WHAT OF WHAT

X Z

WHAT

From what what

Z z z z

Z Z o o o o o

I I

Honors g К a

me> »from φ

it

O u

N K Z

Z oo co

I u o o

I со honor cd cd

ДО 40

With cd it

O o

N Z z

E O co

Z u o ω

it it 04 / it 04 / it 04 / O O O O at O O AT what what WHAT N X of of OF O at ω ^OF N . N N X OF OF OF at OF of of WHAT of of of of O CD O O O O O O O ω O O

it it

04 O O cd o

Co Co

Z z u <1

Z Z Z Z

rH CM WHAT Φ ΙΌ CM CM CM CM CM

WHAT WHAT WHAT

Note:

(a): o-nitrophenylsulfenyl, (b): benzyloxycarbonyl, (c) 1-methyl-2-methoxycarbonylvinyl

Pharmacological evaluation

Methods:

(1) Determination of antimicrobial activity

The minimum inhibitory concentrations (ΜI C, ^ µg / ml) of each test compound were determined by a conventional agar dilution test on an agar broth with cardiac infusion. Each test microorganism was picked with a bacteriological loop (pre-cultivation on trypto-soybean for 16 hours, 10-fold dilution), spread on plates with said agar broth and cultured for 24 hours at 37 ° C.

(2) Determination of acute toxicity

Each test compound was administered introvenously to mice. At 72 hours after administration, a 50% lethal dose [LD 50] was calculated according to the method of Green et al., Brit. J. Pharmacol., 6, 572 [1951].

The results of this pharmacological evaluation are summarized in Tables 1 to 3 below.

Table 1

Antimicrobial efficacy

Test compounds (with reference to the numbers - examples in the description of the invention)

Minimum inhibitory concentration (^ g / ml) Test microorganisms Staphylococcus Pseudomonas aureus 199A aeruginosa Аз

6- [D-2- (DL-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamidio] -penicillanic acid (Example 1)

6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Examples 2, 24, 25 and 27)

6- [D-2- (L-2-amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Example 3)

6- [D-2- (D-2-amino-3-N-isopropylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido) penicillanic acid (Example 4)

6- [D-2- (D-2-amino-3-N-n-butylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Example 5)

6- [D-2- (D-2-amino-3-N-n-hexylcarbamoylpropionamido) -2-p-hydroixyphenylacetamido] penicillanic acid (Example 6)

6- [D-2- (D-2-amino-3-carbamoylpropionamido) -2-p-hydroxyphenylacetamido) penicillanic acid (Examples 7 and 26)

6- (D-2- (DL-2-amino-3-N-methylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid (Example 8)

6- [D-2- (D-2-Amino-3-N-methylcarbamoylpropionamido) -2-phenylacetamide] penicilanoic acid (Examples 9 and 23)

6- (D-2- (L-2-amino-3-N-methylcarbamoylpropionamido) -2-phenylacetamido] penicillanic acid (Example 10)

12.5

3.13

6.25

12.5

12.5

12.5

3.13

12.5

6.25

6.25

12.5

12.5

12.5

12.5

12.5

12.5

12.5

12.5

12.5

Test compounds (with reference to example numbers in the description of the invention) Minimum inhibitory concentration (g g / ml) Test microorganisms Staphylococcus aureus 199A Pseudomonas aeruginosa Аз 6- [D-2- (DL-amino) -S-N-ethylcarbamoyl- 12.5 50 propionamido) -2-phenylacetamido] penicillanic acid (Example 11) 6- [D-2- (DL-2-amino-3-N-n-propylcarb- 50 inoylpropionamido) -2-phenylacetamido ¹ ] penicillanic acid (Example 12) 6- [D-2- (DL-2-amino) -3-Nb utylcarbamoyl- 6.25 50 propionamido) -2-phenylacetamido] penicillanic acid (Example 13) 6- [D-2- (DL-2-amino-3-N- / 3-hydroxyethyl) 12.5 50 1 carbamoylpropionamido) -2-phenylacetamido] penicillanic acid (Example 14) 6- [D-2- (DL-2-amino-3-N-dimethylcarb- 25 50 4 moylpropionamido) -4-phenylacetamido] penicillanic acid (Example 15) 6- [D-2- (DL-2-amino-3-hydroxycarbonyl- 25 propionamido) -2-phenylacttamido] penicillinic acid (Example 16) 6- [D-2- (DL-2-amino-3-methylcarbamoyl- 12.5 12.5 propionamide · ¹ ido) -2-phenylacetamido] penicillanic acid (Example 17) 6- [D-2- (D-2-3-aminoι mlethoxykarbonyl- 12.5 25 Propionamido ¹ ) -4-thienylacetamido] penicillanic acid (Example 18) 6- [D-2- (DL-3-amino-3-hydroxycarbonyl-) 12.5 6.25 propionamido ¹ ) -2-phenylacetamido] penicillanic acid (Example 19) 6- [D-2- (Ы-3-amino-3-methylcarbamoyl- 12.5 50 ^ι propionamido) -2-phenylacttamidO ^1] penicillanic acid (Example 20) 6- [D-2- (DL-3-amino-3-Nn-propylkarba- 12.5 50 moylpropionamido) -2-phenylacetamido] ptnicilanoic acid (Example 21) 6- [D-2- (DL-3-amino-3-N-dimethylcarb- 12.5 25 moylpropionamido) -2-phenylacetamido] penicillanic acid (Example · 22) (Control) Ampicillin 25 > 100 *

Table 2

Antimicrobial efficacy

Test compounds (with reference to the Minimum Inhibitory Concentration (µg / ml) numbers of the Examples in the description of the invention.) Test Microorganisms

Escherichia Pseudomonas coli NIH) JO — 1 cch K — 12 aeruginosa 35R

6- [D-2- [DL-2-Amino-3-N-methylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Example 2)

6- [D-2- (D-2-amino-3-N-methylcarbamoylprapionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Examples 2, 24, 25 and 27)

6- [D-2- (D-2-Amino-3-N-propylcarbamoylpropionamido) -2-p-hydroxyphenylacetamido] penicillanic acid (Example 4)

6- [D-2- (D-2-Amino-3-N-n-butylcarbamoyl propionamido) -2-p-hydroxyphenylaetamide] -penicillanic acid (Example 5)

6- [D-2- (D-2-Amino-3-N-n-hexylcarbamoylpropionamide) -2-p-hydroxyphenylation] -4a-mdo] penicillanic acid (Example 6)

6- [D-2- (D-2-amino-3-carbamoylpropionamido) -2-p-hydroxyphenylacetamide] penicillanic acid (Examples 7 and 26)

6- [D-2- (DL-2-amino-3-N-methylcarbamoylpropionamido) -4-phonylacetamido]] penic acid (Example 8)

6- [D-2- (D-2-amino-3-N-methylcarbamoylpropionamido) -4-phenylacetamido] penicillanic acid (Examples 9 and 23) (Control)

Ampicillin

3.22 3.22 25 3.22 2.56 22.5 6.25 3.22 25 3.22 3.22 25 - - 25 3.22 3Д2 25 6.25 2., 56 50 6.25 2.56 50 6.25 3.22 > 200

Table 3

Acute toxicity

Test compounds (referring to the example numbers in the description of the invention)

Acute toxicity

LD 50, mg / kg (i.v.)

6- [D-2- (D-2-amino-3-N-methylcarbamoyl-> 20,000 propionamido ') -2-p-hydroxyphenylacetamido] penicillanic acid (Examples 2, 24, 25 and 27)

6- [D-2- (L-2-Amino-3-N-methylcarbamoyl-5,475 propionamido) -2-p-hydroxyphenylacetamide] penicillanic acid (Example 3)

6- [D-2- (D-2-amino-3-carbamoylpropion-4,620 amido) -2-p-hydroxyphenylacetamido] penicillanic acid (Examples 7 and 26) 6- [D-2- (D-2- amino-3-N-methylcarbamoyl-2,280 (opionamido) -2-pyrrolidin-2-yl) penicillanic acid (Examples 9 and 23) (Control)> 2,000

Ampicillin

- ^ 7 ^ 202564

Claims (2)

I will invent the subject 1. A process for producing derivatives of 6- (D-2-amino-2-phenylacetamido) penicillanic ^obecn.ého! of formula I CHNH R ^z CH ₃ CH ₃ COQH (!) In which _ R1 represents a hydrogen atom or a hydroxyl group, R2 represents a group of the formula —COCH (NH2) —CH2COR3 or —COCH2 — CH (NH2) —COR3 and R @ 3 denotes a hydroxyl, lower C1 -C6 alkylamino group, a lower C1 -C6 alkylamino group, a lower C1 -C6 alkoxy group or a C1 -C6 hydroxy lower alkyl group, optionally R3 denotes an amino group when R1 is a hydroxyl group or a physiologically acceptable salt thereof, characterized in that 6- (D-2-amino-2-phenylacetamido) -penicillanic acid of the formula II in which: R1 is as defined above and Y1 is hydrogen or a protecting group which is benzyl, phthalimidomethyl, 2,2,2-trichloroethyl or 'phenacyl group, the N-protected amino acid of formula III ^J becného W - OH (III), in which W represents a group of formula -COCH (NHZ) -CH2COR3 or --COCH2 CH (NHZ) -COR 3, R 3 represents optionally, if a hydroxyl group W may be a group of 'the formula -COCH (NHZ) ^2, or --COCH2 -CH2COOY —CH (NHZ) —COO Y2, R3 is as defined above, Y 2 is benzyl; Z denotes a protecting group which is o-nitrophenylsulfenyl, benzyloxycarbonyl or 1-methyl-2-methoxycarbonylvinyl in the presence of diphenylphosphoric azide and at a temperature of 20 to -20 ° C in an inert solvent or a reactive derivative of the compounds of formula III at a temperature from 40 - ^30? C in an inert solvent to give 6- (D-2-acylamido-2-phenylacetamido) penicillanic acid of formula IV wherein R1, Y1 and W are as defined above, the protecting group (s) are removed from compound (IV) and the product is optionally converted into a physiologically acceptable salt. 2. The process of claim 1 wherein the deprotection of the compound (s) from (IV) is carried out when Z is an o-nitrophenylsulfenyl group by treatment with two to three molar equivalents of thiamide at a temperature of 40 to 0 ° C; in the event that Z represents benzyloxycarbonyl and / or Y ¹ Y ² or- benzyl shaken in a hydrogen atmosphere in the presence of palladium on carbonate 'BaCOč barium or palladium on carbon or "palladium black at 40 to 0" C, or, wherein Z is a 1-methyl-2-methoxycarbonylvinyl group by treatment with an aqueous solution of hydrohalic acid, or when Y ¹ is a phenacyl or phthalimidomethyl group, by treatment with an alkali metal salt of a thiol compound or thiophenol at a temperature of -10 to 10 ° C; that Y ¹ denotes a 2,2,2-trichloroethyl group by reduction with formic acid and zinc dust at temperatures 10 to -5 ° C. 1 presses drawings of the north, n., Plant 7, Mast