CS212224B2 - Method of preparation of 7-(substituted benzoylureido) phenylacetamido derivatives of the cephalosporine - Google Patents

Description

The 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- (alpha-substituted benzoylureido) phenylacetamide derivatives of the cephalosporin of formula I,

where

R ₁ is hydrogen or C 1 -C 4 alkyl,

R 1 is C 2 -C 4 alkanoyloxy, n is 2 or 3, wherein at least two of R 1 are attached to adjacent carbon atoms, the position of R 1 is selected from positions 2 to 6, is hydrogen or methoxy and is acetoxyl; S-Rg, wherein Rg is a five-membered heterocyclic ring including one or more nitrogen and optionally sulfur atoms, optionally substituted with C1-C4 alkyl, and pharmaceutically acceptable salts thereof.

Cephalosporin derivatives having a benzoylureido group attached at the alpha position of the 7-acyl amide side chain derivatives are known, for example, from U.S. Patent Nos. 3,925,368 and 4,066,630, British Patent Nos. 1,479,711, 1,498,025, 1,505,885, 1 508,314, i.e. 518,722 and 1,521,073 and DOS 2,653,621. However, none of these patents discloses compounds having a hydroxy 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. 5787/77 cite a lower alkanoyloxy group as substituents, they do not mention a hydroxyl group. However, the above three references provide only a general description of a lower alkanoyloxy group as one of several substituents, and two or three lower alkanoyloxy groups of which at least two are bonded to adjacent carbon atoms are not claimed. Further, no particular compounds having lower alkanoyloxy groups as benzoyl substituents are listed.

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 Fseudomonas or Serratla and the compounds of the invention show a significantly higher antibacterial effect than my cefazorin, cephaloridine or other commonly used cephalosporin antibiotics.

The compounds of the invention have excellent in vivo properties after administration, such as ebsorption, excretion, 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.

Among the compounds of the invention, the compounds of formula I wherein R 1 is C 2 -C 4 alkanoyloxy are not only useful as such, but are also used as intermediates since the alkanoyl group is removed to give a compound of formula I wherein R 1 is hydroxyl.

The alkyl group represented by R ₁ in formula I is C 1 -C 4, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. Preferred examples are methyl and ethyl. The alkanoyloxy group of alkanoyloxy represented by R1 is a group of 2 to 4 carbon atoms which may be branched, which may include, for example, acetyl, propionyl, n-butyryl and isobutyryl. A preferred example is acetyl.

The position of substituent R1 is cell 2 and 3 position, 3 and 4 position, 2, 3 and 4 position, 3, 4 and 5 position, 2, 4 and 5 position, 2, 3 and 5 position, or 2, 3 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 with one or more C 1 -C 4 alkyl groups. 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 such as 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 derivative 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 the formula I and their salts, characterized in that the alpha-ureidobenzylcephalosporin of the formula VII is reacted,

HNCONH -CH -CONH f -S

R, X ₀ - ^ ^Ν γ- ™ 2 ^η *

COOY (VII) wherein R, R ^ and R ^ are as defined above and Y is hydrogen or a protecting group, or a reactive derivative thereof with a benzoyl halide of formula VIII

O ^cox (Λ where Hj is as defined above and Xj is a halogen atom, and optionally is converted to a hydrogen atom, and optionally the compound of formula I obtained is converted into a suitable salt.

(VIII) a substituent Y on a pharmaceutical

The reactive, alpha-ureidobenzylcephalosporin derivative of formula (VII) is a derivative in which the urido group of the acyl group at position 7 is activated to the reactive form.

The reaction is conveniently carried out in a solvent. The solvent to be used in this reaction is an inert organic solvent such as chloroform, dichloromethane, tetrahydrofuran, ethyl acetate, dioxane, acetonitrile and the like. The reaction is conveniently carried out at a temperature of from -10 to 25 ° C, preferably from 0 to Low: 14 ° C. Although the reaction time varies and depends on the reaction temperature and the respective reactants and solvent, it is normally from 1 to 46 hours, preferably from 1 to 10 hours.

Isolation of the compounds of the invention from the reaction mixture can be carried out by any conventional method. For example, extraction 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 and 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 Aoids, 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 various application routes, 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.

212224 4 /

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 upon the body weight of the age, the type of disease and the mode of administration and the frequency of administration.

The process for preparing the compounds of the invention is explained in more detail in the following examples.

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

1. Ethyl acetate-ethanol-acetic acid (25: 5: 1, v / v);

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

Example 1

7- [D - (-) - alpha - (3-methyl-1-ureido) -alpha-phenylacetamido] -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem -4-carboxylic acid (4 g) prepared by reacting one molar equivalent of 7- (D - (-) - alpha-amino-phenylacetamido) -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3 -cephem-4-carboxylic acid with 1.2 molar equivalent of methyl isocyanate was suspended in dichloromethane (80 ml of N, O-bis (trimethylsilyl) acetamide (7.5 ml) was added dropwise to the suspension at 5-10 ° C and the reaction mixture The mixture is stirred at 5-10 ° C until a solution is formed.

To the mixture was added a solution (10 mL) of 3,4-diacetoxybenzoyl chloride (1.9 g) in anhydrous dichloromethane, and the mixture was stirred at room temperature for 5 hours. The reaction mixture is then evaporated to dryness at room temperature under reduced pressure and, after adding anhydrous methanol to the residue, the mixture is again evaporated to dryness under reduced pressure. A mixture of ethyl acetate (150 ml) and cold saturated aqueous sodium bicarbonate solution (200 ml) was added and the mixture was thoroughly mixed under cooling with ice water. After the insolubles were removed, the aqueous phase was separated and the pH adjusted to 1 by adding cold 2N hydrochloric acid. . The precipitate was filtered off, washed with water (100 ml) and dissolved in acetone (100 ml). The solution is treated with activated carbon and the solvent is evaporated off under reduced pressure. The residue is dissolved in a mixture of methanol and chloroform, chromatographed on a silica gel column, and then eluted with the same mixture of solvents. The eluted fractions were collected and the solvent removed by distillation under reduced pressure. The residue was treated with diethyl ether (50 mL) to give 3.5 g of 7- [D - (-) - alpha3- (3,4-di-acetoxybenzoyl) -3-methyl-1-urido) -alpha-phenylaoetamido N -3- (5-methyl-1,3,4-thiazadol-2-ylthiomethyl) -3-oephem-4-carboxylic acid (referred to as compound P below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.50, System II.

iC spectrum:? (? ^-1 ) 3700-2300, 1775, 1690, 1510.

NMR Spectrum: (acetone-d6, 60 MHz) .delta. (Ppm) 2.28 (6H, s), 2.67 (3H, s), 3.15 (3H, s),

3.74 (2H, brs), 4.44 (2H, ABq), 5.12 (1H, d, J = 5Hz), 5.6-6.0 (2H, m), 7.2-7, Δ (8H, m).

(a) 2,3-Diacetoxybenzoyl chloride and 7- [D - (-) - alpha-ureido-alpha-phenylacetamido] -3- (1-methyl-1H-tetrazole-5-) were reacted as described above in this example. ylthiomethyl) -3-cephem-4-carboxylic acid to give 7- [D - (-) - alpha - {3- (2,3-diacetoxybenzoyl) -1-ureido} -alpha-phenylacetamido] -3- (1) methyl-1H-tetrazol-5-ylthiomethyl) -3-oephem-4-carboxylic acid (referred to as compound M below) as a pale yellow powder.

IC spectrum: v (cm ^-1 ) 3700-2300, 1775, 1690, 1530, 1490.

UlcAX

NMR Spectrum: (acetone-d6, 60MHz), .delta. (Ppm) 2.28 ($ H, s), 3.75 (2H, brs), 3.95 (3H, s), 4.4 (2H, brs) 5.07 (1H, d, J = 5 Hz), 5.6-6.0 (2H, m), 7.2-8.0 (8H, m).

(b) 3,4-Diacetoxybenzoyl chloride and 7- [3- (C) -alpha- (3-methyl-1 -ureido) -alpha-phenylaeetamido] -3-acetoxymethyl-3-cephem are reacted as described above. -4-carboxylic acid to give 7- [D - (-) - alpha- {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido] alpha-phenylacetamido] -3-acetoxymethyl-3-cephem -4-carboxylic acid (hereinafter referred to as compound N) in the form of a white powder.

Thin Layer Chromatography: Rf 0.62, System II.

IR: v _{3,700, e} , 2,300, 1,780, 1,745, 1,695, 1,510.

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.00 (3H, s), 2.28 (6H, s), 3.15 (3H, s),

3.5 (2H, broad s), 4.6-5.3 (3H, m), 5.5-6.0 (2H, m), 7.2-7.7 (8H, o (c) 7- [D - (-) - alpha - (3-Methyl-1-ureido) -alpha-phenylaoetamido] -3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3- was reacted as described above. cefem-4-carboxylic acid and 3,4-diacetoxybenzoyl chloride to give 7- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido] -3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-eephem-4-carboxylic acid (referred to as compound R below) as a white powder.

TLC: Rf 0.55, System II.

IR spectrum: _R3 ™ _700-2 300 1775, 1690 1 510th

ifloX

NMR Spectrum: (acetone-d6, 60 MHz) .delta. (Ppm) 2.27 (6H, s), 3.13 (3H, s), 3.75 (2H, brs), 3.93 (3H, s) 4.37 (2H, broad s), 5.11 (1H, d, J = 5 Hz), 5.6-5.9 (2H, m), 7.2-7.7 (8H, m).

(d) Reaction of 3,4-diacetoxybenzoyl chloride and 7beta- [D - (-)

-alpha- (3-methyl-1-ureido) -alpha-phenylacetamido] -7alpha-methoxy-3-acetoxymethyl-3-eephem-4-carbonic acid affords 7beta- [D - (-) - alpha -? 3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido-7alpha-methoxy-3-acetoxymethyl-3-cephem-4-carboxylic acid (referred to as compound Z below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.60, System II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1740-167, 1550.

ιπηχ

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.01 (3H, s), 2.28 (6H, s), 3.17 (3H, s),

3.2 (2H, brs), 3.50 (3H, s), 4.91 (2H, ABq), 5.09 (1H, s), 5.72 (1H, d, J = 7 Hz), 7 2 to 7.7 (8H, m), 8.60 (1H, s), 9.85 (1H, d, J = 7Hz).

(e) 3,4-Diacetoxybenzoyl chloride and 7beta-1D - (-) were reacted as described above.

-alpha- (3-methyl-1 -ureido) -alpha-phenylacetamido] -7 alpha-methoxy-3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid afforded 7beta- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido] -7alpha-methoxy-3- (1-methyl-1H-tetrazole) -5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound AA below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.54, System II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1720-167, 1510.

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.28 (6H, s), 3.16 (3H, s), 3.48 (3H, s),

3.8 (2H, broad s), 3.94 (3H, s), 4.36 (2H, ABq), 4.03 (1H, s), 5.70 (TH, d, J = 7 Hz), 7.2-7.6 (8H, m), 8.6 (1H, s), 9.82 (1H, d, J = 7Hz).

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

Bacillus subtilis PCI-219

2. Staphylococcus aureus 209 P

Staphylococcus aureus JU-5

4. Sarcina lutea B

5. Escherichia coli NIHJ

6. Shigella flexneri 2b

7. Salmonella paratyphi A

Klebsiella pneumoniae 15C

9. Proteus mirabilis 1287

10. Proteus morgani JU-224

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 KAN-2

Pseudomonas aeruginosa Ps-6

18. Serratia Marcescens Ser-25b

19. Serratia márcescens FU-104

20. Enterobacter cloacae FU-250

Table

Tested compound bacteria M N P R OF AA cephaloridine cefazorin 1 3.12 0.4 0.78 0.78 6.25 3.12 Š 0.025 0.20 2 0.78 0.78 0.78 1.56 6.25 1.56 S0.013 0.10 3 6.25 3.12 1.56 3.12 6.25 3.12 0.2 0.78 4 0.4 0.4 0.78 0.78 1.56 0.4 0,025 0.40 5 0.78 0.1 0.2 0.2 0.1 0.1 6.25 1.56 6 1, 56 0.1 0.2 0.2 0.2 = 0.05 1.56 1.56 7 1, 56 0.2 0.2 0.4 0.2 0.1 3.13 1.56 8 6.25 0.2 0.4 0.4 0.4 0.2 3.13 1.56 9 6.25 1.56 3.12 3.12 3.12 1.56 6.25 3.12 10 12.5 0.78 1.56 1.56 1.56 0.78 > 400 100 ALIGN! 11 0.2 0.4 1.56 1.56 1.56 0.4 > 400 > 400 12 0.2 3.12 3.12 3.12 3.12 0.78 > 400 > 400 13 0.1 6.25 6.25 6.25 6.25 1.56 > 400 > 400 14 12.5 0.2 0.4 0.78 0.4 0.1 800 > 800 15 Dec 12.5 0.1 0.2 0.4 0.78 0.1 1, 56 3.12 16 0.2 0.4 1.56 1.56 1.56 0.4 . > 400 > 400 17 0.78 1.56 3.13 3.13 6.25 1.56 > 400 > 400 18 6.25 0.78 1.56 1.56 1.56 0.39 50 400 19 Dec 100 ALIGN! 1.56 1.56 3.13 3.13 0.78 > 400 > 400 20 May 6.25 0.39 0.78 0.39 0.78 0.20 > 400 400

SUBJECT OF THE INVENTION

Claims (4)

1. A process for preparing 7- (substituted alpha-benzoylureido) fenylacetamidoderivétů cephalosporin of formula I (XCON-CONH-CH-CONH-I ^{<R3> R1} N O ^N ^ y " COOH (I) wherein R ₁ is hydrogen or C 1 -C 4 alkyl, R 1 is C 2 -C 4 alkanoyloxy, n is 2 or 3, wherein at least two of R 1 are bonded to adjacent carbon atoms, R 4 is hydrogen or methoxy and R8 is acetoxy or -S-R8, wherein R8 is a five membered heterocyclic ring comprising one or more nitrogen atoms and optionally a sulfur atom optionally substituted by C1-4 alkyl, and pharmaceutically acceptable salts thereof, by the alpha-ureidobenzylcephalosporin of formula VII, HNCONH - CH - CONH AND n. COOY (VII) wherein R p R 6 and R g are as defined above and Y is a hydrogen atom or a deprotecting group, or a reactive derivative thereof is reacted with a benzoyl halide of formula VII wherein R j and n are as defined above and X is a halogen atom; is a protecting group, is converted to a hydrogen atom and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt. 2. The process according to claim 1, wherein the reaction is carried out in an inert organic solvent at a temperature of from -10 to 25 [deg.] C. for 1 to 48 hours. 3. Process according to claim 1 or 2 for the preparation of 7- (alpha-substituovanýoh benzoylureido) fenylaoetamidoderivétů cephalosporin of formula I, referred to in paragraph 1, wherein R, R₁ and n are defined as in point 1, R ₁ is alkyl of 1 to 4 and Rg is an acetoxy or -S-Rg group, wherein Rg is tetrazole substituted with C1-C4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the alpha-ureidobenzylcephalosporin of formula (VII) as set forth in point 1, wherein: R1 and Y are as defined in 1 and R1 and R8 are as defined above, or a reactive derivative thereof is reacted with a benzoyl halide of the formula VIII as defined in 1, wherein R1, X and n are as defined in 1, and when Y is a protecting group, the protecting group 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 preparing the 7- (alpha-substituted benzoylureido) phenylacetamido-cephalosporin derivatives of formula I, as defined in item 1, wherein n is as defined in item 1, R 1 is C 1 -C 4 alkyl; R 6 is hydrogen and R 8 is -S-R 8, wherein R 8 is thiadiazole substituted with C 1 -C 4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the alpha-ureidobenzylcephalosporin of formula (VII) as set forth in point 1, wherein Y is as defined in point 1, and R 1, R 2 and R 8 are as defined above, or a reactive derivative thereof is reacted with a benzoyl halide of the general formula (VIII) as defined in point 1, wherein R 1, X and n are as defined in point 1 and when Y is a deprotecting group, the deprotecting group is converted to a hydrogen atom and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt.