CS212225B2 - Method of preparation of the 7-(substituted benzoylureido)-phenyl-acetamido derivatives of the cephalosporine - Google Patents

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- (alpha-substituted benzoylureido) phenylaetamido-cephalosporin derivatives of formula I,

where

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

R2 is a hydrogen atom or a hydroxy group,

R ₃ is hydroxy, n is 2 or 3, wherein at least two of R 6 are attached to adjacent carbon atoms, the position of R 6 is selected from positions 3 to 5 when R ₁ is alkyl of 1 to 4 carbon atoms, and z positions 2 to 6 when R 1 is hydrogen, R 6 is hydrogen or methoxy and R 6 is acetoxyl or -S-R 8 (where R 8 is a five membered heterocyclic ring including one or more nitrogen atoms and optionally a sulfur atom which is optionally substituted alkylsilyl; C 1 -C 4 carbon atoms and pharmaceutically acceptable salts thereof.

Cephalosporin derivatives having a benzoylureido group bonded at the alpha-position of the 7-acylamide side chain derivatives are known, for example, from U.S. Patent Nos. 3,925,368 and 4,061,630, British Patents δ. 1 479 711, 1 498 025, 1 505 885, 1 508 314, 1 518 722 and 1 521 073 and DOS 2 653 621. However, none of these patents discloses compounds having hydroxy or lower alkanoyloxy as a substituent on the corresponding henzoyl groups. .

Although U.S. Pat. No. 3,687,949, British Pat. No. 1,525,626 and Japanese Patent No. 5787/77 disclose alkanoyloxy as a substituent, not mentioning a hydroxyl group. However, the above three references provide only a general description of the alkanoyloxy group as one of several substituents, and two or three alkanoyloxy groups of which at least two 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 35 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 Pseudomonas or Serratia and the compounds of the invention exhibit 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 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.

The alkyl group represented by R in formula I is a group having 1 to 4 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl. Preferred examples are methyl and ethyl.

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 substituent Ry Therefore, when R 1 is a C 1 -C 4 alkyl, R 3 is either 3 or 4, or 3, 4, or 5. If R 1 is a hydrogen atom, the position of R 3 is either 2 or 3, 3 or 4, 2, 3, or 4, 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 position, the 3 and 4 position or the 3, 4 and 5 position.

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 alkyl groups. Alkyl is a C1-C4 group which may be optionally branched and includes, 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 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 procaine or dibenzylethylenediamine salts. 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 (I) and salts thereof, which comprises removing the protecting groups R 15 and R 21 and / or Y from a protected cephalosporin derivative of formula (IX),

(II) wherein R 1, R 2, K 3 and n are as defined above, R 8 is hydrogen, hydroxy or protected hydroxy, R 8g is hydroxy or protected hydroxy and the position of substituents R 8g is the same as R 3, Y is atom hydrogen or a protecting group, and at least one of R 8, R 6, and Y is a protecting group or is protected, and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt.

In the compound of Formula II, the hydroxyl protecting group for R 8, or R 8, is a protecting group that is readily removed under mild conditions.

Examples of this protecting group are C 2 -C 4 alkenoyl groups such as acetyl, propionyl, butyryl or isobutyryl.

While the removal of this C 2 -C 4 alkenoyl group may be accomplished by any conventional method, it is desirable to perform the removal using a base. The base which is useful in this method is an inorganic or inorganic base. Examples of inorganic bases are ammonium or alkaline salts of weak acids such as carbonic acid, acetic acid, phosphoric acid and the like, ammonium forms of acidic ion exchange resins and moniac. Examples of organic bases are primary, secondary or tertiary amines with one to three alkyl or hydroxyalkyl groups, alicyclic amines such as piperidine or morpholine.

Removal of the protecting group involves treating the protected compound with a base in a solvent in the presence of a compound containing alcoholic hydroxyl groups. In this embodiment, a suitable organic or inorganic base is soluble in the compound with alcoholic hydroxyl groups and in a solvent, preferably a base containing at least one basic nitrogen atom such as ammonia, propylamine, diethylamine, triethylamine, diethylaminomethanol, ethanolamine, triethanolamine, piperidine or morpholine. In addition to these bases, ammonium forms of weak acidic ion-exchange resins may be used in the form of a suspension in a solvent.

Compounds containing alcoholic hydroxyl groups are, for example, alcohols such as methanol, ethanol, ethylene glycol or glycerin, or hydroxy lower alkylamine, such as diethylaminoethanol, ethanolamine or triethanolamine.

A solvent may be used which dissolves the base salt of the protected cephalosporin derivative and the base salt of the cephalosporin derivative of the invention. Translations of these solvents are aprotic polar solvents such as dimethylformemide, non-polar solvents such as dichloromethane and chloroform. Furthermore, of the compounds having alcoholic hydroxyl groups, some may be used as the desired solvents, which compound must meet the solvent requirements as mentioned above. Examples of such compounds are methanol and glycerin.

Preferred combinations for cleavage of the protecting group are methanol-ammonia, triethylamine-triethanolamine-dimethylformamide, diethylaminoethanol-dimethylforoamide and the like. The amount of base may vary and depends on the number of protecting groups in the protected cephalosporin derivative and the total amount of base in the reaction system. The base is typically present in an amount of 1.1 to 30 moles, preferably 3 to 20 moles per mole of protected eephalosporin derivative is used.

Although these compounds are commonly used in large excess as solvents, they can be used in an amount of 2 to 30 moles per mole of protected cephalosporin derivative, when a solvent is also used. In the case of the triethylamine-triethanolamine-dimethylformamide system, one of the preferred systems, triethylamine is normally used in an amount of 0.1 to 10 mol, preferably 2-4 mol, and triethanolamine is commonly used in an amount of 0.7 to 10 mol, preferably 3 to 5 moles per mole of protected cephalosporin derivative. When methanol-ammonia is used, the amount of ammonia ranges from 1.1 to 30 mol, preferably from 2 to 10 mol, per mole of protected cephalosporin derivative and methanol is used in an amount sufficient to dissolve. In the case where the diethyleminoethenol-dimethylformamide system is used, the amount of diethylaminethenol is normally in the range of 2 to 30 mol, preferably 5 db and 20 mol per mol of protected cephalosporin derivative, and dlmethylformamide is used in an amount sufficient to act as a solvent.

Although the reaction temperature varies depending on the use of the compound having an alcoholic hydroxyl group, the appropriate base and the use of a solvent, a temperature in the range of -30 to 40 ° C is commonly used. At least in the case of a compound with hydroxyl groups which also act as a base, the temperature is usually from 10 to 40 ° C, preferably from 20 to 35 ° C, and in the case where the compound with hydroxyl groups also acts as a solvent the temperature is from -30 to 40 ° C, preferably from -25 to 35 ° C.

The reaction time is usually 30 minutes to 20 hours, preferably 1 to 10 hours.

In addition to the deprotection embodiments as explained above, other deprotection techniques from protected cephelosporin derivatives may also be used. One of these techniques is to use a combination of a base containing a nitrogen atom, such as ammonia or a primary or secondary lower alkylamine, with an aprotic solvent, such as dlmethylformamide, capable of dissolving the salt of the cephalosporin derivative with this base.

In another method, a) a base such as an inorganic base such as ammonia, ammonium carbonate, ammonium phosphate, ammonium acetate, sodium vinegar, alkali metal bicarbonate or alkali metal carbonate, organic bases such as ethylamine, diethylamine, trlethylamine, piperidine or morpholine or an ammonium form of a weakly acidic ion exchanger; and b) a solvent such as water or an aqueous mixture of a hydrophilic aprotic solvent such as dioxane.

When the compound of formula IX has a protecting group such as I, examples of protecting groups include groups capable of forming an ester with a carboxyl group such as trimethylsilyl or benzhydryl. The trimethylsilyl group attached to the carboxyl group is readily removed by reaction with water or an alcohol, while the benzhydryl group is readily removed by trifluoroacetic acid.

Other examples of a protecting group for X include organic or inorganic bases capable of forming a salt with a carboxyl group such as alkali metals, alkaline earth metals, and tertiary amines such as trlethylamine, N-methylplperidine, N-methylmorpholine, pyridine and the like. easily removed by acid treatment.

It will be understood that in the case where R 8 ^{and E} 32 3 ^{SO 4} protected groups, the cleavage of these groups is carried out in one way, while the cleavage of the protecting group for Y is carried out in a manner different from that used for R 1 and R 3. If it is necessary not only to remove ^one protecting group Rg ^ use or R g but also the protecting group for Y, the group Y is first removed so that the ring eefamový compound IX remained stable.

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 an β-aminophenylacetic acid or a substituted ureidophenylacetic acid, which is prepared in a conventional manner by reaction, e.g. 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.

For injection, ee may use emulsions, solutions or suspensions in an aqueous or oily carrier. 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 explained in greater detail in the examples.

Thin layer chromatography is carried out in the examples on silica gel 60 Pg54 (finished plates manufactured by E. Merck, Darmstadt) in the following mixtures:

I. Ethyl Ethoethanol-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 triethylemine (6.04 g) in anhydrous dichloromethane (20 ml) was added dropwise at room temperature to a solution of N-methyl-3,4-diacetoxybenzamide (15.0 g) and trimethylsilyl chloride (6.49 g) in anhydrous dichloromethane. (70 mL). After heating the mixture to reflux for 30 minutes, a solution (82 ml) of phosgene (42 ml) in anhydrous dichloromethane is added at -5 ° C to + 5 ° C and the temperature is 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 removal of the insoluble matter by filtration, was used for the next reaction.

2. Add N, O-bis (trimethylstlyl) acetamide (4.3 mL) to a suspension of 7-CD - (-) - alpha-amino-alpha- (4-hydroxyphenyl) acetamido] -3- (1,2, 3-triazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (4.0 g) in anhydrous dichloromethane (50 mL) at room temperature and stirring was continued until mixture was formed.

A solution of 30 ml of N- (3,4-diacetoxybenzoyl) -N-methylcarbamoyl chloride (2.8 g) in anhydrous dichloromethane prepared in Example 1, paragraph 1 is then added dropwise with stirring to the mixture. 10 ° C, the mixture was evaporated to dryness at room temperature under reduced pressure. Anhydrous methanol was added to the residue and the mixture was again evaporated to dryness in vacuo.

Ethyl acetate (150 mL) and cold saturated aqueous sodium bicarbonate solution (150 mL) were added to the residue, and the mixture was stirred vigorously with ice water. After removal of insoluble matter, the aqueous phase is isolated and its pH adjusted to 1 with 2N hydrochloric acid. The precipitate was filtered off, washed with water (100 ml) and dissolved in acetone (100 ml). The acetone solution is treated with activated carbon and the solvent is distilled off under reduced pressure. Treatment of the residue with diethyl ether (100 · 1) sw yields 4.5 g of 7- [D - (-) - alpha-3- (3,4-diecetoxybenzoyl) -3-methyl-1-ureido-alpha- (4-hydroxyphenyl) 3-Amino-3- (1,2,3-triazol-5-yl-thiomethyl) -3-cepein-4-carboxylic acid (referred to as Compound J below) as a pale yellow powder.

Thin Layer Chromatographs: Rf 0.40 (System II)

IR: v (cm ^-1 ) 3700-2300, 1770, 1760, 1680, 16.0.

NMR Spectrum: (acetod-dg, 60 MHz) .delta. (Ppm) 2.26 (6H, e), 3.20 (3H, e), 3.6 (2H, br),

4.1 (2H, br), 5.01 (1H, d, J = 5 Hz), 5.5 to 6.9 (2H, a), 6.7 to 7.7 (7H, m), 7, 86 (1 H, e).

UV spectrum: (EtOH) λ max (run) 268.

a) Following the procedure of Example 1 (2) ee from 7- [D - (-) - alpha-emino-alpha- (4-hydroxyphenyl) acetamido] -3-O-methyl-, 1H-tetrazol-5-ylthiomethyl) - 3-Cephem-4-carboxylic acids prepare. 7- [D - (-) - alpha-3- (3,4-Diaethoxybenzoyl) -3-methyl-1-ureido) -alpha- (4-hydroxyphenyl) abetamido] -3- (1-methyl-, H- tetrazol-5-ylthiomethyl) -3-eephem-4-carboxylic acid (listed as compound to S below).

Also chromatograph on Rf 0.52; System II iC spectrum: v (cm ^-1 ) 3,700 to 2,300, 1,775, 1,690, 1,510.

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.29 (6H, a), 3.17 (3H, b), 3.8 (2H, br), 3.98 (3H, s) 4.40 (2H, broad s), 5.09 OH, d, J = 5 Hz), 5.4-6.0 (2H, m),

6.7 to 7.7 (7H, m).

(b) As described above ee from 7- (D) - (-) - alpha-amino-alpha- (phenylaeetamido) -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3- cephem-4-carboxylic acids 7- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido] -alpha-phenylacetamido] -3- (5-methyl-1,3,4-thiadiezol- 2-ylthiomethyl-3-cephem-4-carboxylic acid (listed as compound P below) as a pale yellow powder.

Thin layer chromatography: Rf 0.50, System II.

IR: v (om ^- ) 3700-2300, 1775, 1690, 1510.

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.28 (6H, s), 2.67 (3H, e), 3.15 (3H, e), 3.74 (2H, brs) 4.44 (2H, ABq), 5.12 OH, d, J = 5 Hz), 5.6-6.0 (2H, m),

7.2-7.7 (8H, m).

(c) Following the procedure described above, from 7- [D - (-) -. alpha.-amino-.alpha .- (4-hydroxyphenyl) acetamide] -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) 3-Cephem-4-carboxylic acids prepare! 7- [D - (-) - alpha - {3- (3,4-diaoethoxybenzoyl) -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 Q below) as a white powder.

Thin-layer chromatography: Rf 0.48, System II.

IR: v (cm @ ^-1 ) 3700-2300, 1775, 1685, 1510.

NMR Spectrum: (acetone-d6, 60 MHz) .delta. (Ppm) 2.28 (6H, s), 2.68 (3H, a), 3.14 (3H, s),

3.65 (2H, broad a), 4.41 (2H, ABq), 5.03 OH, d, J = 5 Hz, 5.5-6.0 (2H, m),

6.6 to 7.6 (7H, m).

(d) Following the procedure described above ee from 7- [D - (-) - alpha-amino-alpha-phenylaeetamido] -3-O-methyl-1H-tetrazol-5-ylthiomethyl) -3-cepea-4-carboxylic acid prepared 7 -fD - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido-alpha-phenylaeetamide] -3- (1-methyl-1H-tetrazol-5-ylthiomethyl) - The 3-cephem-4-carboxylic compound (referred to as compound R below) as a white powder.

TLC: Rf 0.55, System II.

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

NUK spectrum: (acetone d-g, 60 MHz) .delta. (Ppm) 2.27 (6H, s), 3.13 (3H, s), 3.75 (2H, brs),

3.93 (3H, s), 4.3 (2H, broad s), 5.11 (1H, d, J = 5 Hz), 5.6-5.9 (2H, m),

7.2-7.7 (8H, m).

Example 2 (1) Under stirring and cooling, oxalyl chloride (5.3 g) was added to a solution of 2,3-diacetoxybenzamide (4.0 g) in 1,2-dichloroethane (40 ml). The mixture is gradually heated to boiling and allowed to react at this temperature for 10 hours. The solvent and excess oxalyl chloride were distilled off under reduced pressure to give 2,3-diacetoxybenzoyl isocyanate, which was dissolved in anhydrous dichloroethane (40 mL) for the next reaction.

(2) N, O-Bie (trimethylsilyl acetamide (20 mL) was added dropwise to a suspension of 7- (D (-) - alpha-amino-alpha-phenylacetamido) -3- (1-methyl-1H-tetretzol-5-ylthiomethyl)) Of 3-cephem-4-carboxylic acid (12.0 g) in anhydrous dichloromethane (200 mL) at room temperature and stirring was continued until the mixture was dissolved and a solution of the 2,3-diaethoxybenzoyl isocyanate obtained according to Example was added dropwise. The reaction mixture is then stirred for 2 hours at this temperature, evaporated to dryness under reduced pressure and, after adding anhydrous methanol to the residue, the mixture is again evaporated under reduced pressure to dryness. A mixture of ethyl acetate (250 mL) and cold saturated sodium bicarbonate solution (300 mL) was added to the residue, and the mixture was stirred under cooling with ice water, after removal of the insoluble matter, the aqueous phase was separated and adjusted to pH 1 with cold 2N acid. hydrogen chloride . The precipitate was filtered, washed with water (100 ml) and dissolved in acetone (150 mL). The solution is treated with activated carbon and the solvent is distilled off under vacuum. The residue was treated with diethyl ether (100 mL) to give 7 g of 7LD - (-) - alpha - {3- (2,3-di-ethoxybenzoyl) -1-urea} -alpha-phenylacetamido-3- (1- methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound M below as a pale yellow powder).

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

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.28 (6H, s), 3.75 (2H, brs), 3.95 (3H, s),

4.4 C2H, brs), 5.07 (1H, d, J = 5 Hz), 5.6-6.0 (2H, m), 7.2-8.0 (8H, m).

(a) 3,4-Diacetoxybenzoyl chloride and 7beta- [D - (-) - alpha - (3-methyl-1-ureido) -alpha-phenylacetamido] -7alpha-methoxy-3- (1- methyl-1H-tetrazol-5-ylthiomethyl) -3-cephemarboxylic acid to give 7beta- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido N -α-Phenylacetamido-7α-methoxy-3- (1-methyl-1H-tetrazol-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), 5.03 OH, s), 5.70 (1H, d, J = 7 Hz), 7 2 to 7.6 (8H, m), 8.6 (1H, s), 9.82 (1H, d, J = 7Hs).

Example 3

7- [D- (-) - alpha - {3- (3,4-Diaethoxybenzoyl) -3-methyl-1 -ureido} -alpha- (4-hydroxyphenylDacetaoido) -3- (1,2,3-triazole) (5-ylthiomethyl) -3-cephem-4-carboxylic acid (Compound J) (1.2 g), prepared according to Example 1 (2), was dissolved in 10 mL of methanol and methanolic ammonia (0.075) was added dropwise with stirring. g / ml) (3 ml) at -15 to -10 ° C, then stirred at the same temperature for 30 minutes, then poured into cold dilute hydrochloric acid (100 ml) and the precipitate was filtered off and washed The precipitate was dissolved in acetone (100 mL) and the solution was treated with charcoal and evaporated to dryness at room temperature in vacuo, and the residue was treated with diethyl ether (50 mL) to give 0.8 g of 7- [D - (-) -alpha- {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido-alpha- (4-hydroxyphenyl) acetamido] -3- (1,2,3-trlazol-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.

IR spectrum: ν (cm ^-1 ) 3700-2300, 1770, 1680, 1551 nmr spectrum: (DMSO-d6, 60 11Hz) δ (ppm) 3.11 (3K, s), 3.6 ( 2H, brs), 3.95 (2H, bre), 5.03 (1H, d, J = 5 Hz), 5.4 to 5.9 (2H, m), 6.6 to 7.5 (7H, m) 7.95 (1 H, s).

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

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

(a) Following the procedure described above, from 7- [D - (-) -. alpha.- {3- (3,4-diacetosybenzoyl) -3-methyl-1-ure-ldoj-alpha-phenylacetamido] -3- (1- methyl-1H-tetrazol-5-ylthiomethyl) -3-eephem-4-carboxylic acid (compound R) prepared 7- [D - (-) - alpha - [3- (3,4-dihydroxybenzoyl) -3-methyl- 1-ureido-alpha-phenylacetamido] -3- (1-methyl-1H-tetrazol-5-ylthlomethyl) -3-cephem-4-carboxylic acid (referred to as compound E below) as a pale yellow amorphous solid.

Thin Layer Chromatography: Rf 0.40, System II.

IR spectrum: ν (cm ^-1 ) 3700-2300, 1775, 1680, 1551 nmr spectrum: (DMSO-d6, 60 MHz) δ (ppm) 3.10 (3H, s), 3.6 ( 2H, br), 3.93 (3H, e), 4.31 (2H, brs), 5.1 (1H, d, J = 5 Hz), 5.4-5.8 (2H, m), 6 8 to 7.6 (8H, m).

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

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

(b) Following the procedure described above, from 7- [D - (-) - alpha- [3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido] alpha- (4-hydroxyphenyl) ecetemido] -3- (1-methyl-1H-tetrazol-5-ylthlomethyl) -3-cephem-4-carboxylic acid (compound S) affords 7- [D - (-) - alpha-3- (3,4-dihydroxybenzoyl) -3-methyl -1 -ureido-alpha - (4-hydroxyphenyl) acetamido] -3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic compound (referred to as compound P below) as a pale yellow amorphous solids.

Thin Layer Chromatography: Rf 0.39, System II.

IR: in (cm < 3 >) 3,700 to 2,300, 1,770, 1,675, 1,510.

NMR Spectrum: (DMSO-d6, 60 MHz), .delta. (Ppm) 3.11 (3H, s), 3.6 (2H, br), 3.95 (3H, e),

4.31 (2H, brs), 5.1 (1H, d, J = 5 Hz), 5.3-5.9 (2H, m), 6.5-7.5 (7H, m).

UV spectra: (EtOH) λ max (nm) 265, 280, (inflex).

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

(c) Following the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-diethoxybenzoyl) -3-methyl-1-ureido] - alpha -phenylethylamido] -3- (5-methyl- 1,3,4-thiadiezol-2-ylthiomethyl) -3-eephem-4-carboxylic acid (compound P) gives 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3- methyl-1-ureido-alpha-phenylacetamido] -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cefsm-4-carboxylic acid (referred to as Compound C below) as a yellow amorphous solids. Thin Layer Chromatography: Rf 0.40, System II.

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

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.69 (3H, s), 3.19 (3H, s), 3.68 (2H, brs),

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 reactions with ferric chloride *. positive (dark green).

(d) Following the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-diaethoxybenzoyl) -3-methyl-1-ureldo} -elfa- (4-hydroxyphenyl) aeetamldo] -3 - (5-ethyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-eephem-4-carboxylic acid (compound Q) affords 7- [D - (-) - alpha - {3- (3,4) -dihydroxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenyl) acetamido] -3- (5-methyl-1,3,4-thiadiazol-2-ylthio methyl) -3-eephem-4 -carboxylic acid (referred to as Compound D below) as a pale yellow amorphous solid.

Thin-layer chromatography: Rf 0.39, system II.

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

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.70 (3H, s), 3.18 (3H, s), 3.7 (2H, brs),

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).

(e) Using the procedure described above, 7- [D - (-) - alpha - {3- (3,4-diaethoxybenzoyl) -3-methyl-1-ureido} -alpha-phenylae tamido] -3-0,3, 4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid gives 7- [D - (-) - alpha - {3- (3,4-dihydroxybanzoyl) -3-methyl-1-ureido alpha-phenylacetamido -3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound O below) as a pale yellow amorphous solid.

Thin-layer chromatography: Rf 0.40, system II. IR spectrum: v (em ^-1 ) 3700 to 2300, 1775, 1680, 1520.

NMR Spectrum: (DMSO-d6, 60 MHz), .delta. (Ppm) 3.17 (3H, s), 3.7 (2H, br), 4.5 (2H, br),

5.04 (1H, d, J = 5Hz), 5.6-6.0 (2H, m), 6.9-7.7 (8H, m), 9.36 (1H, s).

UV spectrum: (EtOH) λ max (nm) 268.

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

(f) Following the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido] -alpha- (4-hydroxyphenyl) acetamido] -3- (1,3,4-thiadiazol-2-ylthiomethyl) -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-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as Compound H below) in form a pale yellow amorphous solid. Thin-layer chromatography: Rf 0.38, system II.

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

UJBX

N1R spectrum: (DMSO-d6, 60 MHz) δ (ppm) 3.18 (3H, s), 3.7 (2H, brs), 4.5 (2H, br),

5.05 (1H, d, J = 5 Hz), 5.5-6.0 (2H, m), 6.7-7.5 (7H, m), 9.37 (1H, s).

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

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

(g) Following the procedure described above, from 7 - [. D - (-) - alpha - {3- (2,3-diacetoxybenzoyl) -1-ureido] - alpha -phenylacetamido] -3- (1-methyl-1H-tetrazole) -5-ylthiomethyl) -3-eephem-4-carboxylic acid (Compound M) gives 7- [D - (-) - alpha - {3- (2,3-dihydroxybenzoyl) -1-ureido} -alpha-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: v (cm ^-1 ) 3700-2300, 1775, 1680, 1530, 1490.

NMR Spectrum: (acetone-d6, 60 MHz) .delta. (Ppm) 3.7 (2H, br), 3.96 (3H, s), 4.37 (2H, brs), 5.05 (1H, d, J = 5 Hz), 5.6 to 6.0 (2H, m), 6.8 to 7.7 (8H, m).

UV spectrum: (EtOH) λ max (nm) 256, 280 (inflex), 310 (inflex).

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

(h) Following the procedure described above, from 7- [D - (-) - alpha - {3- (3,4-diacetoxybenzoyl) -1-ureido} -alpha-phenylacetamido] -3- (5-methyl-1,3) 4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid gives 7- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -1-ureido} - alpha-phenylacetamiao -3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as the compound)

L below) in the form of a pale yellow powder. Thin-layer chromatography: Rf 0.36, system II

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

NMR Spectrum: (acetone-d6, 60 MHz), .delta. (Ppm) 2.69 (3H, s), 3.72 (2H, bre), 4.4 (2H, br). 5.13 (1H, d, J = 6Hz), 5.5-5.9 (2H, m), 6.8-7.7 (8H,).

UV spectrum: (EtOH) λ max (µm) 269, 290 (inflex).

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

Example 4

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

After stirring at 5-10 ° C for 1.5 hours, 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) was added to the residue and cold 1N hydrochloric acid and the organic phases were 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 solution (700 ml total),

The aqueous phases were separated and washed with ethyl acetate (100 mL). The aqueous phase was adjusted to pH 2.5 with 2M hydrochloric acid and extracted with ethyl acetate (500 mL). The organic phases were 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 - [3- (3,4-diaethoxybenzoyl) -, 3-methyl-1-ureido] -alpha-phenyloetic acid are obtained in the form of a white powder.

TLC: Rf 0.52;

X-ray spectrum: v (cm ^-1 ) 3700 to 2400, 1775, 1740, 1700, 1510.

NMR Spectrum: (DMSO) -dg, 60 MHz) δ (ppm) 2.29 (6H, s), 3.12 (3H, s), 5.35 (1H, d, J = 7Hz), 7.2 to 7.6 (8H, m), 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) phenyloetic acid (6.4 g) prepared by reaction of D - (-) - phenylglyeine with triethylamine (6.4 g) was added dropwise with stirring with methyl isocyanate at a temperature 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-diacetoxyhenzoyl chloride (7.9 g) in tetrahydrofuran (20 ml) was added dropwise and the mixture was stirred at 50 ° C for 2 hours. After cooling to a temperature below 10 ° C, a small amount of methanol was added and the insoluble material 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 Junyaku, K.K. Japan) using 3-4% methanol in chloroform. 5 g of product are thus obtained in the form of a white powder.

(a) The above procedure is repeated except that D - (-) - p-hydroxyphenylglycine is used in place of D (-) - phenylglycine to yield D (-) - alpha- [3- (3,4-diacetoxybenzoyl) -3-Methyl-1-ureido] -alpha- (4-hydroxyphenyl) acetic acid having the following characteristics. Thin Layer Chromatography: Rf 0.51, System I.

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

IDBX

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.2 (6H, s), 3.12 (3H, s), 5.23 (1H, d, J = 7Hz),

6.6 to 7.6 (7H, m), 9.50 (1H, d, J = 7Hz).

Example 5

1. Triethylamine (10.2 g) was added dropwise to a suspension of 7-amino-3-acetoxymethyl-3-cephem-4-carboxylic acid (8.2 g) in anhydrous dichloromethane (80 ml) at room temperature, followed by reaction mixture. Stir 30 minutes at the same temperature. The resulting mixture was used for the following reactions.

2. D - (-) - alpha- [3- (3,4-Diaethoxybenzoyl) -3-methyl-1-ureido] phenylacetic acid (8.6 g) prepared according to Example 4, paragraph 2, and pivaloyl chloride (3, 0 g) was dissolved in anhydrous dichloromethane (40 ml) and triethylamine (2.4 g) was added dropwise at a temperature below -15 ° C.

After stirring at the same temperature for 1 hour, the dichloromethane solution prepared in paragraph 1 above was added dropwise at a temperature below -10 ° C, followed by stirring at the same temperature for up to 1.5 hours. The solution was evaporated to dryness under reduced pressure, and ethyl acetate (200 mL) and cold saturated aqueous sodium bicarbonate solution (200 mL) were added to the residue. After insoluble matter was removed by filtration, the aqueous phase was adjusted to pH 1 with cold 2N hydrochloric acid and extracted with ethyl acetate (200 mL). The isolated organic phase was washed with cold saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue is chromatographed on a Diaion HP-20 resin column (trade name of the resin used for reverse phase chromatography manufactured by Mitsubishi Kasei Kogyo Kabushiki Kalsha, Japan). The methanol-water fractions (3: 1, v / v) were collected and the solvent was distilled off under reduced pressure. 7.0 g of 7- [D - (-) - alpha - {3- (3,4-dlacetoxybenzoyl) -3-methyl-1-ureido] alpha-phenylacetamido] -3-acetoxymethyl-3-cefem-4 are obtained. carboxylic acid (referred to as compound N) in the form of a white powder. Thin Layer Chromatography: Rf 0.62, System II.

iC spectrum: v (cm ^-1 ) 3700-2300, 1780, 1745, 1695, 1510.

NMR Spectrum: (acetone-d _6, 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, m).

(a) Following the procedure of Example 5, 7-amino-3-acetoxymethyl-3-cephem-4-carboxylic acid was reacted with D, (-) - alpha - [-] after slylating with N, O-bis (trimethylsilyl Dacmtamide). 3- (3,4-Diacetoxybenzoyl) -3-methyl-1-ureido] -α-α-hydroxyphenyl) acetic acid, prepared according to Example 4 (a), to give 7- [h - (-) - elfa - {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido} -alpha- (4-hydroxyphenyl) acetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid (hereinafter referred to as compound 0) in the form of white powder. Purification is performed by column chromatography over eilica gel instead of Dieion HP-20 by methanol-chloroform (1:20, v / v). Thin Layer Chromatography: Rf 0.54, System II.

IC spectrum: h. (cm ^-1 ) 3,700 to 2,300, 1,775, 1,740 to 1,670, 1,510.

IDBZ

NMR Spectrum: (acetone-d6, 60 MHz) .delta. (Ppm) 2.00 (3H, s), 2.28 (6H, s), 3.16 (3H, s),

3.5 (2H, brs), 4.6-5.3 (3H, m), 5.5-6.1 (2H, m), 6.7-7.7 (7H, m).

Example 6

2-Diethylaminoethanol (1 mL) was added dropwise to a solution of compound AA (350 g), prepared according to Example 2 (a), in dimethylformamide (1 mL) while cooling with ice water. After stirring at room temperature for 5 hours, the mixture was poured into a mixture of ethyl acetate (50 ml), acetone (10 ml) and 2N hydrochloric acid (50 ml), stirred and cooled with ice water. The organic phase was separated, washed with cold saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The residue was treated with diethyl ether to give 230 mg of 7beta- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido} -alpha-phenylecetamido] -7alpha-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.

Ιδ spectrum: (cm ^_1) of 3 700 to 2 300 1775, 1680 1 515th

max

MUK spectrum: (acaton-dg, 60 MHz) .delta. (Ppm) 3.18 (3H, a), 3.49 <3H, a), 3.5 <2H, ta ··),

3.94 (3H, a), 4.37 (2H, ABq), 5.03 (1H, a), 5.70 (1H, d, J-7Ha), 6.9-

7.7 (8H, a), 8.6 (1H, a), 9.90 (1H, d, J = 7 Hz).

UV Spectrum: (EtOH) λ max (µm) 272, 290 (λflex).

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

(a) Using the procedure of this example, ee from compound N prepared according to Example 5, paragraph (2), prepared 7- [p (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1] [alpha] -alpha-phenylacetamido-3-acetoxymethyl-3-cephem-4-carboxylic acid (referred to as Compound A below) as a pale yellow amorphous solid.

TLC: Rf 0.45, System II.

Ifi spectrum: v (cm ^-1 ) 3700-2300, 1775, 1675, 1520.

NMR Spectrum: (DMSO-d6, 60 MHz) .delta. (Ppm) 2.02 (3H, a), 3.12 (3H, a), 3.47 (2H, brs),

4.7 to 5.2 (3H, m), 5.5 to 5.9 (2H, m), 6.8 to 7.6 (8H, m).

UV spectrum: (EtOH) .lambda.max (nm) -265, 290 (lfflex).

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

(b) Following the procedure described above, Compound 0, prepared according to Example 5 (a), gave 7- [D (-) - alpha - {3- (3,4-dihydroxybenzoyl) -, 3-methyl-1-ureido} .alpha .- (4-hydroxyphenyl) acetamido] -3-acetoxymethyl-3-cephem-4-carboxylic acid (referred to as Compound B below), in the form of a pale yellow amorphous solid fly.

TLC: Rf 0.42, System II.

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

IDBjE

NMR Spectrum: (DMSO-d6, 60 MHz) δ (ppm) 2.01 (3H, s), 3.19 (3H, s), 3.5 (2H, brs),

4.80 to 5.15 (3H, m), 5.5 to 6.1 (2H, m), 6.7 to 7.6 (7H, m).

UV spectrum: (EtOH) .lambda.max (nm) 267, 292 (lfflex).

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

(c) Following the procedure described above, from 7beta- [D - (-) - alpha- {3- (3,4-diacetoxybenzoyl) -3-methyl-1-ureido} -alpha-phenylacetamido-7alpha-methoxy-3- ( 5-Methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-eephem-4-carboxylic acid gives 7beta- [D - (-) - alpha- {3- (3,4-dlhydroxybenzoyl) -3- methyl-1-urea-.alpha.-phenylacetamides -7alpha-methoxy-3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound U) below) 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 reactions with ferric chloride: positive (dark green), (d) Following the procedure described above, from 7beta- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-urea to 7-.alpha.-Phenylaoetam-7-.alpha.-methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid yields 7beta- [D - (-) - .alpha.-{ 3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido-.alpha.-phenylaetamido] -7 .alpha.-methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylo acetic acid (referred to as compound V) in the form of a pale yellow powder.

Thin Layer Chromatography: Rf 0.52, System II.

IR spectrum: ^ r _(Cb -1) _{3700 until 2300> 775)} 6 ₈ 0, 520th

UloX

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

(e) Following the procedure described above for 7beta- [D - (-) -. alpha.- {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureldo} -alpha- (4-hydroxyphenyl) acetamido-7alpha-methoxy -3- (1-methyl-1H-tetrazol-5-ylthioraethyl) -3-cephem-4-carboxylic acid gives 7beta- [D - (-) - alpha-.beta .- (3,4-dlhydroxybenzoyl) 13

2,2225

3-Methyl-1-urea-D-alpha- (4-hydroxyphenylacetamido) -7alpha-methoxy-3- (1-methyl-1H-tetrazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid (referred to as compound) W below) in the form of a light-colored powder.

Thin Layer Chromatography: Rf 0.49) System II.

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

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

(f) Following the procedure described above, from 7beta- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1-ureido] alpha - (4-hydroxyphenyl) acetamido] -7alpha- methoxy-3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid gives 7beta- [D - (-) - alpha-3- (3,4- dihydroxybenzoyl) -3-methyl-1-urea-.alpha .- (4-hydroxyphenyl) -a-acetamide-7alpha-methoxy-3- (5-methyl-1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem -4-carboxylic acid (referred to as compound X below) as a pale yellow powder.

Thin Layer Chromatography: Rf 0.50, System II.

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

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

(g) Following the procedure described above, from 7beta- [D - (-) - alpha - {3- (3,4-dihydroxybenzoyl) -3-methyl-1 -ureido} -alpha- (4-hydroxyphenyl) acetamide: 7 alpha-Methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid gives 7beta- [D - (-) - alpha - {3- (3,4- dihydroxybenzoyl) -3-methyl-1-ureido [alpha] - (4-hydroxyphenyl) acetamido-7α-methoxy-3- (1,3,4-thiadiazol-2-ylthiomethyl) -3-cephem-4-carboxylic acid ( reported as compound X below) as a pale yellow powder.

Thin-layer chromatography: Sf 0.50, system II.

IR: v (cm ^-1 ) 3700-2300, 1775, 1680, 16.0.

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

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 to 6 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 mlrabilis, 287, 0. Proteus morganl JU-244

11. Pseudomonas aeruginosa J-272

12. Pseudomonas aeruginosa J-169

13. Pseudomonas aeruginosa J-169-CM222.4. Pseudomonas aeruginosa GNB-75

Pseudomonas aeruginosp GNB-75-M57740

16. Pseudomonas aeruginosa KAN-2

Pseudomonas aeruginosa Ps-6

18. Serratia marcescens Ser - 25b, 9. Serratia marcescens FU-104

20. Enterobacter cloacae FU-250

Table

Tested compound bacteria AND (B) C D E ' F G 1 0.4 0.4 0.4 0.78 0.4 0.78 0.4 2 0.78 0.78 0.4 0.78 0.4 0.78 0.2 3 3.12 6.25 3.12 3.12 3.13 3.13 1.56 4 0.78 0.78 0.4 0.4 0.2 0.2 0.2 5 á 0,1 W0.1 šo, 1 50.1 0.1 50.1 áo, 1 6 -0.1 ž 0,1 SO, 1 50.1 SsO, 1 50.1 50.1 7 áo.i - 0,1 ao.i So.i -0.1 50.1 50.1 8 0.2 0.2 so, i 0.4 0.1 0.1 50.1 9 0.78 0.78 0.4 0.78 0.4 0.78 0.4 10 1.56 1.56 0.4 0.78 0.4 0.4 0.4 11 0.4 0.2 0.4 6.85 0.78 0.78 0.78 12 3.12 1.56 1.56 1.56 1.56 1.56 1.56 13 3.12 3.12 3.12 6.25 3.12 3.12 6.25 14 á 0,1 0.2 0.2 0.78 0.39 0.78 0.4 15 Dec ž 0,1 áo, 1 á 0,1 0.4 Ξο, ι 50.1 50.1 16 0.4 0.2 0.4 78; 0.78 0.78 0.78 17 0.78 0.78 0.78 1.56 0.78 1.56 0.78 18 0.78 1.56 0.4 0.78 0.4 0.78 0.4 19 Dec 1.56 6.25 0.78 1.56 0.78 1.56 0.78 20 May 0.4 0.78 0.4 1.56 0.78 1.56 0.78

Table

Tested compound bacteria H AND TO L T W cephaloridine cefazorln 1 0.76 0.76 3.12 1.56 3.12 3.12 0,025 0.20 2 0.4 0.78 1, 56 0.78 1, 56 3.12 0.013 0.10 3 3.22 3.12 6.25 3.12 1.56 1.56 0.2 0.78 4 0.4 0.4 0.78 0.4 0.4 0.4 0,025 0.40 5 0.39 50.1 0.78 0.78 £ 0.05 S0.05 6.25 1.56 6 50.1 50.1 1.56 1, 56 Ž0,05 = 0.05 1.56 1.56 7 50.1 ao.i 1.56 0.78 Sř0,05 S0.05 3.13 1.56 8 0.2 50.1 6.25 6.25 0.1 0.1 3.13 1.56 9 0.78 50.1 6.25 25 1.56 3.12 6.25 3.12 10 0.4 0.2 12.5 25 0.39 0.39 > 400 100 ALIGN! 11 1, 56 3.12 0.2 1, 56 0.4 0.4 > 400 > 400 12 0.78 24 0.2 6.25 0.78 0.78 > 400 > 400 13 6.25 50 0.1 6.25 0.78 0.78 > 400 > 400 14 0.4 0.78 6.25 1.56 0.1 0.2 800 > 800 15 Dec 0.2 50.1 12.5 0.1 S0.05 = 0.05 1.56 3.12 16 0.78 1, 56 0.2 1.56 0.4 0.4 > 400 > 400 17 0.78 6.25 0.78 6.25 0.78 1.56 > 400 > 400 18 0.78 0.4 6.25 50 0.20 0.4 50 400 19 Dec 1, 56 0.78 100 ALIGN! 200 0.39 0.39 > 400 > 400 20 May 3.13 0.4 6.25 50 0.20 0.4 > 400 400

Claims (13)

SUBJECT OF THE INVENTION CLAIMS 1. A process for the preparation of 7- (alpha-substituted benzoylureido) phenylecetamidoderivatives of cephalosporin of the formula I wherein R1 is hydrogen or C1-C4 alkyl, Rg is hydrogen or hydroxy, ^R 3 is hydroxy, n is 2 or 3, wherein at least two of R Ej ^D ° u attached to adjacent carbon atoms of substituents R is selected from positions 3 to 5, where R is alkyl having 1 to 4 carbon atoms and from 2 to 6 when R 1 is hydrogen, R 6 is hydrogen or methoxy, and R 6 is acetoxy or -S-R 8, wherein R 8 is a five-membered heterocyclic ring including one or more nitrogen atoms, and optionally a sulfur atom which is optionally substituted by a C 1 -C 4 alkyl group and pharmaceutically acceptable salts thereof, characterized in that the protecting groups R 6 and R 8 and / or Y are removed from the protected cefelosporin derivative of the general formula II, wherein R 4 is R 6, R 6a and not as defined above, R 6 is hydrogen, hydroxy or protected hydroxy, R 6g is hydroxy or protected hydroxy and the position of substituents such as R 6, Y is hydrogen or protecting group and at least one of R 8, R 8, and Y is a protecting group or is protected and optionally the compound of formula I obtained is converted into a pharmaceutically acceptable salt. 2. The method of claim 1, wherein the protected cephalosporin derivative is a compound wherein R8, R8 and R8 are independently lower alkanoyloxy. 3. A process according to claim 2 wherein the protected cephalosporin derivative is reacted with an organic or inorganic base. 4. The process of claim 3 wherein the inorganic base is a base selected from the group consisting of ammonium or alkali salts of carbonic acid, acetic, phosphoric, ammonium forms of weakly acidic ion exchange resins and ammonia, and the organic base is a base selected from groups including primary, secondary or tertiary alkyl · amines, primary, secondary or tertiary hydroxy-lower alkylamines, and alicyclic amines such as piperidine and morpholine. 5. A process according to claim 3, wherein the reaction with the base is carried out in the presence of a compound with an alcoholic hydroxyl group or groups in a solvent. 6. A process according to claim 5, wherein the base is a compound having one or more nitrogen atoms and the compound is an alcohol or a hydroxyalkylamine and the solvent is a non-protonic polar or non-protonic non-polar solvent. 7. A process according to claim 6, wherein triethylamine is used as the base, triethenolamine as said hydroxyl compound and dimethylformamide as the solvent. 8. A process according to claim 6, wherein ammonia is used as the base and methanol is used as the hydroxyl compound and the solvent is methanol. 9. The process of claim 6 wherein the base and the hydroxyl group are diethylaminoethanol and dimethylformamide is the solvent. 10. The process according to items 1 to 9 for the preparation of 7- (alpha-substituted benzoylureido) phenylacetamido-cephalosporin derivatives of the general formula 1 as defined in item 1, wherein R8 and n are as defined in item 1, R is alkyl of 1 to 4 carbon atoms , R 6 is methoxy, R 8 is -S-R 8, wherein R 8 is 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: removing the protecting group R ^ ^and g ^ 21 ^and / ^{net) 0 1} protected derivative ^of cephalosporin of formula II shown in item 1, wherein R ,, ^ 32 '^ ^{8 n} as defined in paragraph 1 and R ,, R and R 8 are as defined above, and optionally converting the obtained compound of formula X to a pharmaceutically acceptable salt. 11. A process according to items 1 to 9 for the preparation of 7- (alpha-substituted benzoylureldo) phenylacetamide derivatives of cephalosporin of general formula I as defined in item 1, wherein R 1, R 8, R 8 and n are as defined under 1. R 8 is acetoxy or -S-R 8, wherein R 8 is thiadiazole, optionally substituted with C 1 -C 4 alkyl, or trlazole, and pharmaceutically acceptable salts thereof, characterized in that the protecting groups R 32 and R 8 are removed and / or Y from a protected cephalosporin derivative of the general formula Ii, as defined in 1, wherein R 1, R 8, R 8, R 8, Y and n are as defined in point 1 and R 6 and R 8 are as defined above; to a pharmaceutically acceptable salt. 12. The process according to items 1 to 9 for the addition of 7- (alpha-substituted benzoylureido) phenylaeetamidoderivatives of the cephalosporin of formula I, as defined in item 1, wherein R8, R3 and n are as defined in item 1, R1 is hydrogen; hydrogen and Rg is -S-Rg, wherein Rg is tetrazole substituted with C1-C4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the protecting groups R-jg and R ₂ are removed and / or Y from a protected cephalosporin derivative of the general formula II as defined in item 1, wherein R 8, R 8, R 8, Y and n are as defined in 1 and R 1, R 3 and R 8 are as defined above; a pharmaceutically acceptable salt. 13. A process according to 1 to 9 for the preparation of 7- (alpha-substituted benzoylureido) phenylaetoetido derivatives of the cephalosporin of formula (X), as defined in (1), wherein R1, R8, R1 and n are as defined in (1). R 8 is -S-R 8 wherein R 8 is tetrazole substituted with C 1 -C 4 alkyl, and pharmaceutically acceptable salts thereof, characterized in that the protecting groups R 6 and R 8 are removed and / or Y from the protected cephalosporin derivative of the general formula II, as defined in 1, wherein R 1, R 8, R 8, R 8, Y and n are as defined in point 1 1 and R 6, R 8 are as defined above, and optionally the compound of formula I obtained is converted to a pharmaceutically acceptable salt.