CS212226B2 - Method of preparation of 7-(substituted benzoylureido)- phenyl-acetoamido derivatives of the cephalosporine - Google Patents

Abstract

The invention relates to cephalosporin derivatives having a broad antibacterial spectrum and they exhibit antibacterial effect on various gram-negative and gram-positive bacteria. More specifically, the invention relates to 7- (β-substituted-benzoylureido) -phenylacetamide derivatives the cephalosporin of formula (I) where is R1 is hydrogen or alkyl of 1 to 4 carbon atoms, x R 8 is hydrogen or hydroxy, R6 is hydroxy, n number 2 or 3 taking. at least two of R 1 are attached to adjacent carbon atoms, position R 1 is selected from positions 3 to 5 when R 1 is C 1 -C 4 alkyl, and from positions 2-6 when R 1 is hydrogen, R 1 is hydrogen or methoxyl and R 8 is 5-membered a heterocyclic ring including one or more nitrogen atoms and optionally a sulfur atom, which is optionally substituted by C 1 -C 4 alkyl and pharmaceutically acceptable suitable salts.

Description

The present invention relates to cephalosporin derivatives having a broad antibacterial spectrum and exhibiting antibacterial activity against various gram-negative and gram-positive bacteria.

More specifically, the invention relates to 7- (β-substituted-benzoylureido) phenylacetamido derivatives of cephalosporin of formula I,

where is

R 1 is hydrogen or C 1 -C 4 alkyl, _x

R 8 is a hydrogen atom or a hydroxy group,

R @ 1 is hydroxy, n is the number 2 or 3, wherein. at least two of the substituents R 1 are attached to adjacent carbon atoms, the position of the substituent R 1 is selected from positions 3 to 5 when R ₁ is alkyl of 1 to 4 carbon atoms, and positions 2 to 6 when R 1 is hydrogen, R R 6 is hydrogen or methoxy and R 8 is a five-membered heterocyclic ring including one or more nitrogen and optionally sulfur atoms optionally substituted with C 1 -C 4 alkyl and pharmaceutically acceptable salts thereof.

Cephalosporin derivatives having a benzoylureldo group attached at the alpha position of the 7-acylaidide side chain derivatives are known, for example, from U.S. Patent Nos. 3,925,368 and 4,061,630, British Patent Nos. 1,479,711, 1,498,025, 1,550,885, 1,508 However, neither of these patents disclose compounds having a hydroxyl or lower alkanoyloxy group as a substituent on the corresponding benzoyl group.

Although U.S. Patent No. 3,687,949, British Patent No. 1,525,626, and Japanese Patent No. 5,950,176. . No. 5787/77 disclose alkanoyloxy as a substituent, not mentioning a hydroxyl group. However, the three references above provide only a general description of the alkanoyloxy group as one of several substituents, and two or three lower alkanoyloxy groups, at least two of which are bonded to adjacent carbon atoms, are not claimed. The following are no specific compounds having alkanoyloxy groups as benzoyl substituents.

As explained above, the cephalosporin derivatives of the invention of formula (I) are novel.

The derivatives according to the invention show a high antibacterial effect against both Gram-positive and Gram-negative bacteria. In particular, they are effective against bacteria belonging to Peeudomonas or Serratia. The compounds of the invention exhibit a considerably higher antibacterial effect than cefazorin, cephaloridine or other commonly used cephalosporin antibiotics.

The compounds of the invention have excellent properties in vivo application, such as absorption, secretion, distribution, metabolism and the like, and have the ability to prevent infection by bacteria. Because of these properties, the compounds of the invention are useful as antibacterial agents.

When R 1 is a C 1 -C 4 alkyl group and the hydroxyl group is present on the benzoyl nucleus at the 2 or 6 position, the ureido group becomes unstable, thus limiting the position of the R 1 j substituent. Therefore, when R 1 is alkyl, the position of R 1 is in the 3 and 4 positions or 3, 4 and 5. For the other combinations of R 1 and R 1, the position of R 1 is b and the 2 and 3 positions are 3 and 4 respectively; 4 position, 3, 4 and 5 position, 2, 4 and 5 position, 2, 3 and 5 position, or 2, and 6 position. Preferred are the 2 and 3 positions, the 3 and 4 positions or the 3, 4 and 5 positions.

The five-membered heterocyclic group represented by Rg of formula I includes, for example, 1,3,4-thiadiazole, triazole and tetrazole, which may optionally be substituted by one or more alkyl groups of 1 to 4 carbon atoms. Alkyl is a group having 1 to 4 carbon atoms which may be optionally branched and include, for example, methyl, ethyl, n-propyl and isopropyl. Preferred alkyl is methyl.

Since the cephalosporin derivatives of the invention have a carboxyl group, they are capable of forming salts with various alkaline compounds. All such salts are also within the scope of the invention. Examples of salts of the compounds of the invention are inorganic alkali salts, for example, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium salts and salts with organic bases such as salts with procaine or dibenzylethylenediamine. These salts can be prepared in a conventional manner, i.e. by reacting the free carboxyl group of the cephalosporin derivatives with the inorganic or organic base described above.

Due to the asymmetric carbon in the 7-acetamido group, the end products of the invention are optical isomers, i.e., DL-, D- and L-isomers. All such isomers are also within the scope of the invention.

The present invention provides a process for the preparation of cephalosporin derivatives of formula 1 and salts thereof, characterized in that the 3-acetoxymethylcephalosporin of formula II,

R.

'1

R.

CON-CONH

WITH

ch ₂ -o-coch _{3 (W} wherein R 1, R 8, R p and R 8a are as defined above and X is hydrogen or a protecting group, is reacted with a thiol or a salt thereof of formula III,

(III) wherein R 8 is as defined above, and when X is a protecting group, X is converted to a hydrogen atom and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt.

In this reaction, the thiol is generally used in the form of the sodium or potassium salt. The thiol in free form may be used in the presence of an inorganic base such as an alkali metal hydroxide, an alkali metal carbonate, an alkali metal bicarbonate and the like or in the presence of an organic base such as a trialkylamine and the like.

The reaction is conveniently carried out in a solvent such as acetone, methanol, ethanol, tetrahydrofuran and the like, and their aqueous solutions may also be used.

The reaction is generally carried out with heating or at room temperature. That is, re-? the operating temperature is usually in the range of 20 to 70 ° C, preferably 45 to 55 ° C. The reaction time is from 5 to 60 hours, preferably from 5 to 30 hours.

Isolation of the compounds from the reaction mixture is accomplished by any conventional method. For example, extraction is performed with an organic solvent, such as dichloromethane, chloroform or ethyl acetate, and chromatography on silica gel, ion exchange resin, crosslinked dextran, high-porosity styrene polymer or acrylic ester or the like.

Optical isomers, such as the D- or L-isomers of the compounds of the invention, can be prepared using an optically active starting compound, such as alpha-aminophenylacetic acid or substituted ureidophenylacetic acid, which is prepared by conventional resolution methods, for example as described in JP Greenstein and M Winitz in Chemistry of the Amino Acids, Vol. 1, pp. 715-760, John Wiley and Sons, N.Y. (1961).

The compounds of the invention may be formulated into various pharmaceutical formulations prepared for a variety of methods of administration, in the manner described for other cephalosporin compounds.

The invention also relates to pharmaceutical compositions for the treatment of humans and animals. The preparation is carried out using conventional pharmaceutical carriers, diluents and additives.

Emulsions, solutions or suspensions in an aqueous or oily carrier may be employed for injection. Suppositories are prepared using conventional suppository ingredients such as coconut oil or glycerides.

The content of active compound varies and varies depending on the mode of administration.

It is commonly used above 0.1%, such as 5 to 99%, preferably 10 to 60%.

The amount administered for an adult is between 100 and 3000 mg per day. Preferably, an amount in the range of 500 to 2000 mg per day is used and is dependent on body weight, age, disease type and method of administration and frequency of administration.

The process for preparing the compounds of the invention is further illustrated in the Examples.

Thin-layer chromatography is carried out in the examples on silica gel 60-254 (metal plates manufactured by E. Merck, Darmstadt) in the following mixtures:

I. ethyl acetate-ethanol-acetic acid (25: 5: 1, v / v),

II. ethyl acetate-ethanol-acetic acid-water (10: 4: 2: 1, v / v).

Example 1

1. A solution of triethylamine (6.04 g) in anhydrous dichloromethane (20 ml) was added dropwise to a solution of N-methyl-3,4-diacetoxybenzamide (15.0 g) and triethylsilyl chloride (6.49 g) in anhydrous dichloromethane at room temperature. (70 ml). After refluxing the mixture for 30 minutes, a solution (82 ml) of phosgene (42 ml) in anhydrous dichloromethane was added at -5 to +5 ° C and the temperature was allowed to gradually rise to room temperature. Excess phosgene and solvent were evaporated under reduced pressure to give crude N- (3,4-diacetoxybenzoyl) -N-methylcarbamoyl chloride. The product was dissolved in cold anhydrous dichloromethane (50 mL) and after insoluble matter was removed by filtration, it was useful for the next reaction.

2. Add N, O-bis (trimethylsilyl acetamide (44.3 mL) to a suspension of D - (-) - phenylglycine (14.0 g) in anhydrous dichloromethane (150 mL) at room temperature and stir the reaction mixture until complete. A solution of N- (3,4-dieoethoxybenzoyl-N-methylcarbamoyl chloride in anhydrous dichloromethane obtained in step 1) was added dropwise with stirring.

After stirring for 1.5 hours at 5-10 ° C, the mixture was evaporated to dryness at room temperature and after addition of methanol to the residue, the mixture was evaporated again to dryness under reduced pressure at room temperature.

Ethyl acetate (500 mL) and cold 1 H hydrochloric acid were added to the residue, and the organic phase was separated. The organic phase was washed with cold saturated aqueous sodium chloride solution (500 mL) and extracted three times with cold saturated aqueous sodium bicarbonate in a total volume of 700 mL.

The aqueous phase was separated and washed with ethyl acetate (100 mL). The aqueous phase was adjusted to pH 2.5 with 2N hydrochloric acid and extracted with ethyl acetate (500 mL).

The organic phase was separated and washed with cold saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate and evaporated to dryness. 15.0 g of D (-) - alpha-.beta .- (3,4-diacetoxybenzoyl-3-methyl-1-ureido) -alpha-phenylacetic acid are obtained in the form of a white powder: TLC: System I: Rf 0.52

IR: in cm ^-1 3700 to 2400, 1775, 1740, 1700, 1510.

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.29 (6H, s), 3.12 (3H, s), 5.35 (1H, d, J = 7 Hz),

7.2-7.6 (8H, a), 9.65 (1H, d, J = 7Hz).

The above compound was also prepared as follows. To a mixture of tetrahydrofuran (50 ml), trimethylsilyl chloride (7 g) and D (-) - alpha - (3-methyl-1-ureido) phenylacetic acid (6.4 g) prepared by reaction of D (-) - phenylglycine with methyl isocyanate is added dropwise. from stirring triethylamine (6.4 g) below 10 ° C. After the addition was complete, the mixture was stirred at 40-50 ° C for one hour, then cooled to below 10 ° C. A solution of 3,4-dieoethoxybenzoyl chloride (7.9 g) in tetrahydrofuran (20 mL) was added dropwise to the mixture, followed by stirring at 50 ° C for two hours.

After cooling to a temperature below 10 ° C, a small amount of methanol was added to the mixture and the insoluble matter was filtered off. The filtrate was evaporated to dryness at room temperature under reduced pressure and the residue was purified by silica gel column chromatography (Wakogel C-200, manufactured by Wako Junyeku, K.K. Japan) using 3-4% methanol in chloroform. 5 g of product are obtained in the form of a white powder.

3. To a solution of D - (-) - alpha - [3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido] phenyloetic acid (2.0 g) in methanol (20 mL) was added dropwise with ice cooling. water 29% aqueous ammonia solution (4 mL). The mixture was stirred for 30 minutes, and the temperature was allowed to slowly rise to room temperature, then the reaction mixture was concentrated at room temperature under reduced pressure. Ethyl acetate (100 mL) and cold saturated aqueous sodium bicarbonate solution (70 mL) were added to the residue, and the pH of the aqueous solution was adjusted to 8.5.

The aqueous phases were separated and after adjusting the pH to 2.5 with cold 2N hydrochloric acid, extraction was performed with ethyl acetate (200 mL). The organic phase is separated, washed with cold saturated aqueous sodium chloride solution, dried over anhydrous sodium sulphate and evaporated to dryness under reduced pressure to give 1.3 g of D (-) - alpha - [3- (3,4-dihydroxybenzoyl) -3 methyl-1-ureido] phenylacetic acid as a white powder.

4. D (-) - [alpha] - [3- (3,4-Dihydroxybenzoyl) -3-methyl-1-ureido] -alpha-phenylacetic acid (4.0 g) dried over phosphorus pentoxide, dissolved in anhydrous tetrahydrofuran ( 60 ml) containing 1-hydroxybenzotriazole (1.56 g). To the solution was added dropwise 20 ml of a solution of Ν, Ν'-dicyclohexylcarbodiimide (2.87 g) in anhydrous tetrahydrofuran at 0 ° C under cooling with an ice bath and a nitrogen atmosphere. Stirring is continued and the temperature is allowed to slowly rise to room temperature over 2 hours and the resulting precipitate, Ν, Ν'-dicyclohexylurea is filtered off. The filtrate containing D (-) - alpha - [3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] phenylacetic acid 1-benzotriazolylester is useful for the next reaction.

5. Add N, O-bis (trimethylsilyl) acetemide (11.5 mL) under a nitrogen atmosphere at room temperature to a suspension of 7-amino-3-acetoxymethyl-3-oephem-4-carboxylic acid (6.32 g) in anhydrous dichloromethane (100 mL) and then stirred until complete dissolution. The solution prepared in paragraph 4 in tetrahydrofuran was added dropwise while maintaining the temperature at 5-10 ° C during the addition, followed by stirring for 8 hours. The mixture was then evaporated to dryness at room temperature and, after adding anhydrous methanol to the residue, re-evaporation under reduced pressure.

Ethyl acetate (100 mL) was added to the residue, and cold saturated aqueous sodium bicarbonate solution (150 mL) was added to the residue, and the mixture was thoroughly mixed. After insoluble matter was removed by filtration, the pH of the filtrate was adjusted to 1.0 with 2N hydrochloric acid. The precipitate was filtered off, washed with water (100 ml) and dissolved in acetone (70 ml). The solution is treated with activated carbon and evaporated to dryness under vacuum at room temperature.

Acetone (5 mL) was added to the residue, and the mixture was treated with diethyl ether (50 mL).

3.5 g of 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] -phenyl-phenylacetamido] -3-acetoxymethyl-3-cefem-4 are obtained. -carboxylic acids (referred to as Compound A below) as a pale yellow amorphous solid.

TLC: Rf 0.45, System II.

IR spectrum: _{3 700-2} 300 1775, 1675 1 520th

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.02 (3H, s), 3.12 (3H, s), 3.47 (2H, brs), 4.7-5.2 ( 3H, m), 5.5-5.9 (2H, m), 6.8-7.6 (8H, m).

UV spectrum: (EtOH) λ max (nm) 265, 290 (inflex).

Color reaction with ferric chloride: positive (dark green).

(a) The procedure described above is repeated except that intermediate D (-) - alpha - [3- (3,4-diaoethoxybenzoyl) -3-methyl-1-ureido] - alpha - (4-hydroxyphenyl) acetic acid has the following characteristics:

Thin Layer Chromatography: Rf 0.51, System I.

IR: v (cm ^-1 ) 3700-2300, 1770, 1740, 1700-167, 1510.

NMR Spectrum: (DMSO-d6, 60 MHz) [delta] (ppm) 2.2 (6H, s), 3.12 (3H, s), 5.23 (1H, d, J = 7 Hz), 6.6 to 7.6 (7H, m), 9.50 (1H, d, J = 7Hz) to give 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3 methyl-1-ureido-N- (4-hydroxyphenyl) acetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid (referred to as Compound B below) as a pale yellow amorphous solid.

TLC: Rf 0.42, System II.

Spektrδ spectrum: in (cm ´) 3,700 to 2,300, 1,775, 1,675, 1,510.

NMR Spectrum: (DMSO-d6, 60 MHz), < 5 (ppm) 2.01 (3H, s), 3.19 (3H, s), 3.5 (2H, bre), 4.80-5. 15 (3H, m), 5.5-6.1 (2H, m), 6.7-7.6 (7H, a),

UV spectrum: (EtOH 1) λ max (nm) 267, 292 (inflex).

Color reaction with ferric chloride positive (dark green).

Example 2

7- [D - (-) - alpha - {3- (3,4-Dihydroxybenzoyl) -3-methyl-1-ureido} -afarphenylecetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid (compound A) (0.57 g) prepared according to Example 1, paragraph 5, is suspended in a phosphoric acid buffered solution of pH 6.3 (10 mL). Sodium bicarbonate (0.70 g) was dissolved in the suspension and then 5-methyl-2-mercapto-1,3,4-thiadiazole (0.13 g) was added. The solution is left for 24 hours, maintaining the pH between 6.0 and 6.5 with dilute hydrochloric acid and sodium bicarbonate, and maintaining the temperature between 45 and 55 ° C.

The reaction mixture was cooled, washed with ethyl acetate and the aqueous phase separated. The pH of the aqueous phase is adjusted to 1.0 with dilute hydrochloric acid and the precipitate is filtered off. The precipitate was washed with water (50 mL) and dissolved in acetone (20 mL). The solution was treated with activated carbon and evaporated to dryness under reduced pressure at room temperature.

The residue was treated with diethyl ether (10 mL) to give 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha-phenylecetamido] -3- ( 5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound C below) as a pale yellow amorphous solid.

Thin Layer Chromatography: Rf 0.40 (System II).

Ιδ spectrum: (cm ´) 3,700 to 2,300, 1,775, 1,680, 1,515.

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.69 (3H, s), 3.19 (3H, s), 3.68 (2H, bre), 4.4 (2H, br) 5.04 (1H, d, J = 5 Hz), 5.6-6.1 (2H, m), 6.9-7.7 (8H, m).

UV spectrum: (EtOH) λ max (nm) 272.

Color reaction with ferric chloride: positive (dark green).

(a) Following the procedure described above, from 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} - alpha - (4-hydroxyphenyl) acetamido] -3 -acetoxymethyl-3-cephem-4-carboxylic acid gives 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} - alpha - (4-hydroxyphenyl) acetamido -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound D below) as a pale yellow amorphous solid.

Thin Layer Chromatography: Rf 0.39, System II.

Spektrδ spectrum: in (cm ´) 3,700 to 2,300, 1,775, 1,680, 1,510.

Nuclear Magnetic Resonance Spectrum (DMSO-d6, 60 MHz) δ (ppm) 2.70 (3H, a), 3.18 (3H, a), 3.7 (2H, bre), 4.4 (2H, br), 5.05 (1H, d, J = 5 Hz), 5.5-6.0 (2H, m), 6.7-7.5 (7H, m).

UV spectrum: (EtOH) λ max (nm) 270, 280 (inflex).

Color reaction with ferric chloride *. positive (dark green).

(to) Following the procedure described above, from 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido] -3-acetoxymethyl-3-cephem -4-carboxylic acid: 7- [D (-) - alpha- [3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha-phenylecetamido] -3- (1-methyl-1H-) tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound E below) as a pale yellow amorphous substance.

Thin Layer Chromatography: Rf 0.40, System II.

Spektrδ spectrum: in (cm ´) 3,700 to 2,300, 1,775, 1,680, 1,515.

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 3.10 (3H, s), 3.6 (2H, br), 3.93 (3H, s), 4.31 (2H, brs) 5.1 (1H, d, J = 5 Hz), 5.4-5.8 (2H, m), 6.8-7.6 (8H, m).

OT spectrum: (EtOH) λ max (nm) 265, 285 (inflex).

Color reaction with ferric chloride: positive, (dark green) t

(c) Following the procedure described above, from 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} - alpha - (4-hydroxyphenyl) acetamido] -3- acetoxymethyl-3-cephem-4-carboxylic acid affords 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] alpha - (4-hydroxyphenyl) acetams to give -3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound F below) as a pale yellow amorphous solid.

Thin layer chromatography: Rf 0.39, System II.

IR spectrum: in (cm &lt; 3 &gt;) 3,700 to 2,300, 1,770, 1,675, 1,510. '

NMR Spectrum: (DMSO-d6, 60 MHz), s (ppm) 3.11 (3H, s), 3.6 (2H, br), 3.95 (3H, s), 4.31 (2H, brs) 5.1, 1H (d, J = 5 Hz), 5.3-5.9 (2H, m), 6.5-7.5 (7H, m).

OT spectrum * (EtOH) λ max (nm) 265, 280 (inflex), 290 (itlex).

Color reaction with iron chloride team: positive (dark green).

(d) Following the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido-1-alpha-phenylaeetamido] -3-acetoxymethyl-3- cefem-4-carboxylic acid: 7- [D (-) -. alpha .- {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido] -3- (1,3,4) thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound G below) as a pale yellow amorphous solid.

Thin Layer Chromatography: Rf 0.40, System II.

IR: (cm -1) 3,700 to 2,300, 1,775, 1,680, 1,520.

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 3.17 (3H, s), 3.7 (2H, br), 4.5 (2H, br), 5.04 (1H, d, J = 5 Hz), 5.6-6.0 (2H, m), 6.9-7.7 (8H, m), 9.36 (1H, s).

OT spectrum: (EtOH) .lambda.max (nm) 268.

Color reaction with ferric chloride: positive (dark green).

(e) the procedure hereinbefore described of 7- [D (-) - a- {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -elfa- (4 ^J! hydroxyphenyl) acetamido] -3vacetoxymethyl -3-cephem-4-carboxylic acid gives 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenyl) acetamido] - 3- (1,3,4-thiediezol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound H below) as a pale yellow amorphous solid.

Thin Layer Chromatography: Rf 0.38, System II.

IR: (cm ^-1 ) 3700-2300, 1775, 1680, 1510.

NMR Spectrum: (DMSO-d6, 60 MHz), s (ppm) 3.18 (3H, s), 3.7 (2H, brs), 4.5 (2H, tSr), 5.05 (1H, d) , J = 5 Hz), 5.5 to 6.0 (2H, m), 6.7 to 7.5 (7H, m), 9.37 (1H, s). "

OT spectrum: IStOH) λ max (nm) 270, 280 (inflex).

Color reaction with ferric chloride: positive (dark green).

(f) Following the procedure described above, from 7- [D- (D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenylacetamido) - 3-acetoxymethyl-3-cephem-4-carboxylic acid gives 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenyl) acetamido-3- (1,2,3-triazol-5-ylthiomethyl) 3-cephem-4-carboxylic acid (referred to as Compound I below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.39, System II.

ifi spectrum: (cm “) 3,700 to 2,300, 1,770, 1,680, 1,515.

NMR Spectrum: (DMSO-d6, 60MHz), δ (ppm) 3.11 (3H, s), 3.6 (2H, brs), 3.95 (2H, brs),

5.03 (1H, d, J = 5Hz), 5.4-5.9 (2H, m), 6.6-7.5 (7H, m), 7.95 (1H, s).

OT spectrum: (EtOH) λ max (nm) 272,280 (inflex).

Color reaction with ferric chloride: positive (dark green).

(g) Following the procedure described above, from 7- [D - (-) -. alpha.- {3- (2,3-dihydroxybenzoyl) -1-ureido] -alpha.

-phenylaeetmido] -3-acetoxymethyl-3-eephem-4-carboxylic acid and 1-methyl-5-mercapto-1H-tettazole gives 7- [D - (-) - alpha - {3- (2,3-dihydroxybenzoyl)] -1-ureido} -α-phenylacetamido-3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound K below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.38, System II.

IR spectrum: in (cm -1) 3,700 to 2,300, 1,775, 1,680, 1,530, 1,490.

NMR Spectrum: (acetone-d6, 60 MHz) δ (ppm) 3.7 (2H, br), 3.96 (3H s), 4.37 (2H, brs),

5.05 (1H, d, J = 5Hz), 5.6-6.0 (2H, m), 6.8-7.7 (8H, m).

UV spectrum: (EtOH) λ max (nm) 256, 280 (inflex).

Color reaction with ferric chloride: positive (dark green).

(h) Using the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -1-ureldo} -alpha

-phenylacetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid and 5-methyl-2-mercapto-1,3,4-thiadiazole gives 7- [D (-) - alpha - {3- (3,4) -dihydroxybenzoyl) -1-ureido-alpha-phenylacetamido -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound L below) in the form of light of yellow powder.

Thin Layer Chromatography: Rf 0.36, System II.

IR: v (cm ^-1 ) 3700-2200, 1770, 1675, 1525.

NMR Spectrum: (acetone-d6, 60 MHz) δ (ppm) 2.69 (3H, s), 3.72 (2H, brs), 4.4 (2H, br),

5.13 (1H, d, J = 6Hz), 5.5-5.9 (2H, m), 6.8-7.7 (8H, m).

UV spectrum: (EtOH) λ max (nm) 268, 290 (inflex).

Color reaction with ferric chloride: positive (dark green).

(i) Following the procedure described above, from 7-beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] -alpha-phenylacetamido] -7alpha-methoxy-3-acetoxymethyl- Of 3-cephem-4-carboxylic acid and 1-methyl-5-mercapto-1H-tetrazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] - alpha-phenylacetamido] -7-alpha-methoxy-3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound T below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.51, System II.

IR: (cm ^-1 ) 3700-2300, 1775, 1680, 1515.

NMR Spectrum: (acetone-d6, 60 MHz) δ (ppm) 3.18 (3H, s), 3.49 (3H, s), 3.5 (2H, brs),

3.94 (3H, s), 4.37 (2H, ABq), 5.03 (1H, s), 5.70 (1H, d, J = 7Hz), 6.9 to 7.7 (8H) m, 8.6 (1H, s), 9.90 (1H, d, J = 7Hz).

UV spectrum: (EtOH) λ max (nm) 272, 290 (inflex).

Color reaction with ferric chloride: positive (dark green).

(j) Following the procedure described above, from 7beta- [D - (O-alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] -alpha-phenylacetamido] -7alpha-methoxy-3-acetoxymethyl Of 3-cephem-4-carboxylic acid and 5-methyl-2-mercapto-1,3,4-thiadiazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl -1-urido-alpha-phenylacetamido] -7alpha-methoxy-3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound U below) in in the form of a pale yellow powder.

Thin Layer Chromatography: Rf 0.52, System II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1680, 1515.

Color reaction with ferric chloride: positive (dark green).

(k) Following the procedure described above, from 7beta- [D (-) - alpha - {3- (3,4-hydroxybenzoyl) -3-methyl-1-ureido] -alpha-phenylacetamido] -7alpha-methoxy-3-acetoxymethyl Of 3-cephem-4-carboxylic acid and 2-mercapto-1,3,4-thiadiazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1 -ureidoj] -alpha-phenylacetamido] -7alpha-methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound V) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.52, System II.

IR: ^v (cm ^-1 ) 3700-2300, 1775, 1680, 1520.

Color reaction with ferric chloride: positive (dark green).

(1) Following the procedure described above, from 7beta-.alpha .- (-) - alpha-.beta. Of 3-cephem-4-carboxylic acid and 1-methyl-5-mercapto-1 H -tetrazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-, ureido 7-β-Methoxy-3- (1-methyl-1H-tatrazol-5-ylthiomethyl) -3-oephem-4-carboxylic acid (referred to as compound W below) of yellow powder.

TLC: Rf 0.49, System II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1675, 1510.

Color reaction with ferric chloride: positive (dark green).

(m) Following the procedure described above, from 7beta- [D (-) - alpha- {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenylacetamido) -7alpha-methoxy-3 of acetoxymethyl-3-cephem-4-carboxylic acid and 5-methyl-2-mercapto-1,3,4-thiadiazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3- methyl-1-ureido-alpha- (4-hydroxyphenyl) acetamido-7-alpha-methoxy-3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid ( reported as compound X below) as a yellow powder.

Thin layer chromatography: Rf 0.50, IT system.

IR spectrum: ν (cm ^-1 ) 3700-2300, 1775, 1680, 1510.

Color reaction with ferric chloride: positive (dark green).

(n) Following the procedure described above, from 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenylacetamido) -7alpha-methoxy-3 of acetoxymethyl-3-cephem-4-carboxylic acid and 2-mercapto-1,3,4-thiadiazole gives 7beta- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3 * methyl-1 -ureido} - alpha - (4-hydroxyphenyl) acetamido] -7-alpha-methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound Y below) ) in the form of a yellow powder.

Thin Layer Chromatography: Rf 0.50, System II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1680, 1510.

Color reaction with ferric chloride: positive (dark green).

The minimum inhibitory concentration (MIC in µg / ml) for some species of bacteria is determined for the compounds of the invention prepared according to Examples 1 and 2 and the results are summarized in the following table. In the table, the bacteria are numbered according to the following data.

Bacilus subtilis PCI-219

2. Staphylococous aureus 209 P

Staphylococcus aureus JU-5

4. Sarcina lutea B

5. Escherichia coli NIHJ 6. Shigella flexneri 2b

7. Salmonella paratyphi A

Klebsiella pneumonise 15C

9. Proteus mirabilis 1287

10. Proteus morgani JU-244

11. Pseudomonas aeruginosa J-272

12. Pseudomonas aeruginosa J-169

Pseudomonas aeruginosa J-169 CM222

14. Pseudomonas aeruginosa GNB-75

Pseudomonas aeruginosa GNB-75-M57740

16. Pseudomonas aeruginosa ΚΑΝ-2

Pseudomonas aeruginosa Ps-6

18. Serratia marcescens Ser-25b

19. Serratia marcescens FU-104

20. Enterobacter cloacae FU-250.

Tested compound bacteria C D E F O H 1 1 0.4 0.78 0.4 0.78 0.4 0.78 0.78 2 0.4 0.78 0 4 0.78 0.2 0.4 0.78 3 3.12 3.12 3.13 3.13 1.56 3.12 3.12 4 0.4 0.4 0.2 0.2 0.2 0.4 0.4 5 <0.1 £ 0.1 0.1 £ .0.1 £ 0.1 0.39 £ 0.1 6 <0.1 £ 0.1 £ 0.1 40.1 £ 0.1 40.1 £ 0.1 7 0,1 0,1 <0.1 <0.1 £ 0.1 Sat 1 <o, 1 £ 0.1 8 <0.1 0.4 0.1 0.1 £ ° »i 0.2 £ 0.1 9 0.4 0.78 0.4 0.78 0.4 0.78 £ 0.1 10 0.4 0.78 0.4 0.4 0.4 0.4 0.2 11 0.4 6.25 0.78 0.78 0.78 1.56 3.12 12 1.56 1.56 1.56 1.56 1.56 0.78 24 13 3.12 6.25 3.12 3.12 6.25 6.25 50 14 0.2 0.78 0.39 0.78 0.4 0.4 0.78 ’ 15 Dec <OJ 0.4 £ 0.1 £ 0.1 £ 0.1 0.2 £ 0.1 16 0.4 0.78 0.78 0.78 0.78 0.78 1.56 17 0.78 1.56 0.78 1.56 0.78 0.78 6.25 18 0.4 0.78 0.4 0.78 0.4 0.78 0.4 19 Dec 0.78 1.56 0.78 1.56 0.78 1.56 0.78 20 May 0.4 1.56 0.78 1.56 0.78 3.13 0.4

Test compound

bacteria TO L T W OF AA oefa- loridine cefa- zorin 1 3.12 1.56 3.12 3.12 6.25 3.12 = 0.025 0.20 2 1.56 0.78 1.56 3.12 6.25 1.56 = 0.013 0.10 3 6.25 3.12 1.56 1.56 6.25 3.12 0.2 0.78 4 0.78 0.4 0.4 0.4 1.56 0.4 0,025 0.40 5 · 0.78 0.78 <0.05 £ 0.05 0.1 0.1 6.25 1.56 6 1.56 1.56 £ 0.05 £ 0.05 0.2 £ 0.05 1.56 1.56 7 1.56 0.78 £ 0.05 £ 0.05 0.2 0.1 3.13 1.56 8 6.25 6.25 0.1 0.1 0.4 0.2 3.13 1.56 9 6.25 25 1.56 3.12 3.12 1.56 6.25 3.12 10 12.5 25 0.39 0.39 1.56 0.78 > 400 100 ALIGN! 11 0.2 1.56 0.4 0.4 1.56 0.4 > 400 > 400 12 0.2 6.25 0.78 0.78 3.12 0.78 > 400 > 400 , 3 OJ 6.25 0.78 0.78 6.25 1.56 > 400 > 400 14 6.25 1.56 0.1 0.2 0.4 0.1 800 > 800 15 Dec 12.5 0.1 = 0.05 = 0.05 0.78 0.1 1.56 3.12 16 0.2 1.56 0.4 0.4 1.56 0.4 > 400 > 400 17 0.78 6.25 0.78 1.56 6.25 1, 56 > 400 > 400

242226 continued table

Tested product bacteria TO L T W OF AA cefa- loridine cefa- zorin 18 6.25 50 0.20 0.4 1.56 0.39 50 400 19 Dec 200 200 0.39 0.39 3.13 0.78 > 400 > 400 20 May 6.25 50 0.20 0.4 0.78 0.20 > 400 400

SUBJECT OF THE INVENTION

Claims (4)

SUBJECT OF THE INVENTION A process for the preparation of 7- (alpha-substituted benzoylureido) phenylacetamide derivatives of oephalosporin of the general formula I, wherein: R @ ₁ is hydrogen or C1 -C4 alkyl, R @ ₂ is hydrogen or hydroxy, R 1 is hydroxy, n number 2 or 3, wherein at least two of R 1 are bonded to adjacent carbon atoms and the position of R 1 is selected from positions 3 to 5 when R 1 is alkyl of 1 to 4 carbon atoms, and position 2 up to 6 when R 6 is a hydrogen atom, R 6 is hydrogen or methoxy and R 8 is a five membered heterocyclic ring containing one or more nitrogen atoms and optionally a sulfur atom optionally substituted by C 1 -C 4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the 3-acetoxymethylcephalosporin of formula II wherein R ₁ , R ₂ , R 1, R 2 and n are as defined above and Y is a hydrogen atom or a protecting group, reacted with a thiol or a salt of formula III, HS-Rg (III) wherein Rg is as defined above, and when Y is a protecting group, Y is converted to a hydrogen atom and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt. 2. The process of claim 1, wherein the reaction is carried out at a temperature of 20 to 70 ° C for 5 to 6 hours in a solvent selected from the group consisting of acetone, methanol, ethanol and tetrahydrofuran. 3. The process according to items 1 and 2, for the preparation of 7- (alpha-substituted benzoylureido) phenylacetamide derivatives of cephalosporin of general formula 1, as mentioned in item 1, wherein R 1, R 8, R 8a and n are as defined in 1 and R 6 is and R 8 is triazole, thiadiazole optionally substituted with C 1 -C 4 alkyl, and tetrazole substituted with C 1 -C 4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the 3-acetoxymethylcephalosporin of formula (II) referred to in point 1 wherein R, Rg, Rj, Y and n are as defined in 1 and R 1 is as defined above, reacted with the thiol or a salt thereof of the general formula (III) as defined in item t, where R g is as defined above, and Y is a protecting group, Y is converted to a hydrogen atom, and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt. 4. The process of items 1 and 2 for the preparation of 7- (alpha-substituted benzoylureido) phenylacetamide derivatives of the cephalosporin of formula I as set forth in 1, wherein R 1, R 6, R 6 and n are as defined in 1 and R 1 is hydrogen and R 6 is triazole, thiadiazole, optionally substituted with C 1 -C 4 alkyl, or tetrazole substituted with C 1 -C 4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the 3-acetoxymethylcephalosporin of formula (II) above is provided in R ₁ , R 8, R 8, Y and n are as defined in 1 and R 1 is as defined above, reacted with a thiol or a salt of formula III as defined in 1, wherein R 3 is as defined above, and Y is a protecting group, Y is converted to a hydrogen atom, and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt.