SU1250173A3 - Method of producing derivatives of cephalosporin - Google Patents

Abstract

Novel cephalosporin derivatives of the formula <IMAGE> wherein R<1> is hydrogen or a conventional amino-protecting group, and R<2> is a straight or branched alkyl group containing from 1 to 4 carbon atoms, allyl, 2- butenyl or 3-butenyl, or is a group <IMAGE> wherein R<3> and R<4> each are independently hydrogen, methyl or ethyl, or R<3> and R<4>, taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and nontoxic pharmaceutically acceptable salts, physiologically hydrolyzable esters and solvates thereof exhibit potent antibacterial activity and can be incorporated into pharmaceutical compositions.

Description

This invention relates to a process for the production of new cephaloporin derivatives, which. are antibiotics and can be used in medical practice.

The purpose of the invention is the creation of new antibiotics of the cephalosporin series, which have increased activity and low toxicity.

 . , ..:

Example 1. 7- (g) -2-methoxy-imino 2- (2-aminothiazol-A-yp) acetamido to-3- (1-methyl-1-pyr-olidinium) IU type -3-cephem-A carboxylate (b).

A. Benehyde 3-chloromethyl ester L-7- (g) -2-Mexoxyimino -2- (2-tritylaminothiazol-yl) acetamido - -3-cephem-4-carboxylic acid Gy.

3-Chloromethyl-7-amino-3-cephem-4-carboxylic acid benzhydryl ester (2.29. G, mmo) in (57 ml) is treated with bic- (trimethylsilyl) acetamide (BCA, 4.09 nl 16.6 mmol) at a temperature of 50 minutes for obtaining a clear solution. An acid chloride solution, prepared from (2) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetic acid (2.04 g, 4.60 mmol), and PClj ( 1.15 g, 5.52 mmol) in methylene chloride (20 ml), the mixture is stirred at room temperature for 30 minutes, poured into cold water (200 ml) and extracted with ethyl acetate (3 x 10 ml), combined, the extracts are washed with aqueous NaCl, dried and evaporated. The remaining syrup (4 g) is chromatographed on silica gel (150 g) 6 column by elution with 10: 1 and 3: 1 mixture of toluene and ethyl acetate successively (successively. The fractions containing the desired mixture are combined and evaporated to give 2 , 61 g (68%) of compound VI in the form of amorphous powder. NMR, S gDCl, ppm: 3.50 (2H, s); 4.0- (3N, s); 4.33 (2H, s); 4.96 (1H, d); 5.87 (1H, KB); 6.65 (1H, s); 6.90 (1H, s); 7.3 (25H, m).

B. Benzhydryl ester of 3-iodomethyl-7-j (Z) -2-methoxyimino-2- (2-tritylaminothiazol-4-yl) acetamido-3- -cepheme-4-carboxylic acid.

The mixture of the 3-chloromethylsilicate of Example 1A (1.50 g, 1.79 mmol) and NaJ (1.34 g, 8.93 mmol) in methyl ethyl ketone (30 ml) was stirred at room temperature for 1 hour.

After the solvent is removed, the residue is dissolved in ethyl acetate (100 ml) and washed with water, aqueous 8.8. and aqueous NaCl, dried and evaporated with

obtaining the target compound

(1.47 g, 89%) as an amorphous powder.

NMR (JcDClj, 3, 55 (2H, AVQ.); 4.00 (3N, s); 4.25 (2H, s);

4.97 (1H, d); 5.80 (1H, q), 6.65 (1H, s); 6.90 (1H, s); 7.3 (25H, m). C. 7- (E) -2-methoxyimino-2- (2-aminothiazol-4-.yl) acetamido-3- (1-methyl-1-pyrrolidinium) methyl 1-3-cephem-.

5-4-carboxylate (1a) ..

A mixture of the compound according to Example 1B (4.5 g, 4.93 mmol) and H-methylpyrro. lidine (0.65 ml, 6.20 mmol). (45 ml) was stirred at room temperature for 20 minutes. Ether (300 ml) is added to the mixture to isolate a quaternary salt of blocked cephalosporin, which is collected, filtered, and treated with 90% trifluoroacetic acid (TFA) (40 ml) at room temperature for 1 hour. Then the mixture is evaporated at reduced pressure and temperature below 20 ° C. The residue was triturated with ether to give the TFA salt of compound a (2.40 g), which was dissolved in methanol (5 ml) and treated with a 1% solution of sodium-2-ethylhexoate (NEEG) in ethyl acetate (8 ml)

5 at room temperature for 30 minutes. After adding ethyl acetate (100 mp), the resulting precipitate (1.94 g) is collected by filtration. A high pressure liquid chromatography (HPLC) analysis shows that the crude product is 7 / J purity with a ratio between the A-isomer and -isomer of 1: 8. Purification of the product by HPLC was repeated three times (Likhrosorb RP-18, 3-300 mm),. elute with 5% aqueous CHjOH or. 0.01 M ammonium phosphate buffer (pH 7.2) containing 5% CHjOH to obtain 35 mg (1.5%) of the desired product as a colorless powder. Estimated purity (ghvd) 90%. T.Sh1. (with decomposition). . IR spectrum (KVg) 1770; 1660; 1620, 55, UV spectrum, „„ “phosphate buffer, pH 7) and (2): 235 (16200); 258 (15400).

NMR, D, 0, 2.31 (4H, m); 3.08 (ЗН, s); 3.63 (4H, m); 4.09

0

(ZN, s); 5.43 (1H,, and 4.8 Hz), 5.93 (1H, d), 7.08 (1H, s).

Pp and. M e p 2. 7- (2) -2-methoxy 1mio-2- (2-amino-thiaool-4-yl) acetates (1-methyl-1-pyrrolidinium) methyl 3-3-cephem-4 -carbroxylate (1a).

To a stirred solution of 20.4 g (21.9 mmol) of the compound of example 1B, in 150 ml of dry methylene chloride, was added 2.42 g (28.5 mmol) of 1-methylpirrrlidine, in one portion of HFN at room temperature. The mixture is stirred for 5 minutes and poured into 1000 m of ether with vigorous stirring from equipment 4 precipitates, which is filtered, washed with ether (5 x 30 nl) and dried in vacuum to obtain 19.3 g of the blocked product as a pale yellow powder .

IR spectrum, (KBG), 1m0 “l, cm: 3400; 1780 (s), 1740, 1675/1530.

TLC: solvent (ethanol / CHC1 (1: 3), Rf 0.30, (Rf = 0.95 for compound VIIa).

The solid is dissolved in 185 ml of TFA / water (99: 1), stirred for 1 hour at room temperature, and concentrated to 30 ml at a temperature below 10 seconds. The concentrate is poured into 1000 ml of EtF at. vigorous, stirring to form, which is filtered off, washed with ether 5x 40 ml) and dried in vacuum to obtain 10.6 g of a pale yellow powder. The powder is dissolved in 20 ml of methanol and the solution is filtered. To the filtrate is added 45 ml of 0.8 and NEEG IB ethyl acetate. The resulting suspension is poured onto 400 ml of ethyl acetate and filtered to obtain 8.08 g of solid matter, which is a mixture of the desired compound and the corresponding l-isomer () according to the HPLC analyte (LIHROSB RP-18, 10-15% methanol in 0.01 M phosphate buffer, pH 7). A second run of 28.9 g (31.0 mmol) of compound VII6 gave 16.0 g of the crude product (1: 8). Isolation of the desired d-eomer from the combined crude product (24.08 g) using preparative HPLC (System 500, Waters Associates, PropPAC 500 / Cj, 5-10C CHjOH) yields 769 mg of Compound 1c.

Example 3. 7-1 (g) -2-methoxy-imino-2- (2-aminathiazl-4-yl) acetami2501734

Aoj-3- (1-methyl-1-pyrrolidinium) me-. Tyl 3-Z-cephem-4-carboxylate (Ta).

A series of experiments were conducted to determine the effect of the amount of solvent and the reaction time on the compound 1 ° and the ratio aV d in the reaction product. The general technique is as follows.

To a suspension of the 3-iodomethyl derivative of Example 1B (45 mg, 0.048 mmol) in the specified amount of the indicated solvent was added a solution of N-methylpyrrolcdine (0.01 ml, 0.097 1 mmol) in ether (0.1 ml) and the mixture was stirred at room temperature for a specified period. The reaction mixture was diluted with ether (5 ml) and the resulting precipitate was collected by filtration and mixed Ayu.t with 90% TFA. The mixture is stirred for Hr and evaporated to dryness under reduced pressure at a temperature below to give the product. The ratio for the product is determined by HPLC (Ichrocop6, mobile phase 0.1 M ammonium phosphate buffer, pH 7.2, containing 15% CHjOH, retention time min, D 5.56 min). 30 Product yield and isomer ratios for each experiment are given in Table. one.

PRI me R 4. 7- (g) -2-ethoxy5 imino-2- (2-aminothiazol-4-yl) acetates, up to 1-3-3 (1-methyl-1-pyrrolidine) methyl-3- 3-cephem-4-carboxylate (j).

A. Benzhydryl ester of 3-chloromethyl-7- (g) -2-ztoxyimino-2- (2-tri® tilaminothiazol-4-yl) acetamido-3-cephem-4-carboxylic acid.

To a solution of (g) - 2-ethoxymino-2- - (2-trisch1-amino-thiazol-4-yl) acetic acid (1.095 g, 2.4 mmyl) in dichloromethane (20 ml) is added phosphorus chloride (500 Mg ). After stirring for 1 hour at room temperature, the mixture is added in one portion to the cooled

on ice to a solution of 7-amino-3-chloromethyl-3-cephem-4-carboxylic acid benzhydryl ester (1.083 g, 2.4 mmol) and BSA (1 ml) in dichloromethane (20 ml), after stirring

5 for half an hour, the reaction mixture was poured into IOZ-aq aqueous NaHCOj (200 ml) and extracted with CHCl j (100.ML). Extract washed

512501

One day, dried MgSO4 and evaporated under reduced pressure.

The residue is chromatographed on a silica gel column. CHCl elution,

J

ate said compound as j

morphic powder, t, 76 g (86%).

NMR, ((CDClI), 1.40 (3N, .CH.CHj); 3.55 (2H, ABca, 2-CH,); 37 (2H, s, -CHjO); 4.60 (2H, q, CH, CH,); 4.90 (1H, g, 6-H); 5.89 10 (1H, g, 7-H); 6.88 {1H, s, H thiazole;, 91 (1H , s, CH benzhydryl).

B. Diphenylmethyl 7- (Z) -2-ethoxyimino-2- (2-tritylaminothia-. Ol-4-yl) acetamido-3-iodomethyl-3-15-cephem-4-carboxylic acid ester.

A mixture of the compound from example 4A (1.07 g, 1.25 mmol) and NaJ (562 mg, 2.75 mmol) in acetone (20 ml) was stirred for 1 hour. The mixture was filtered. 20, the filtrate was poured into water and ek. stratified with ethyl acetate. The organic layer is sequentially washed with 5% aqueous, water and saturated aqueous NaCl, dried over. 25 MgS04 and evaporated to obtain 1.04 g (89%), the specified connection

NMR, CDCl ,, 3.55 (2H, q, 2-CH); 4.27 (2H, s, CHj-1); 5.02, (1H, d, 6-H); 5.87 (1H, d, 7-H); 30

6.68 (1H, s, H thiazole ring), 6.93 (1H, s. SI benzhydryl).

C. 7- (Z) -2-ethoxymin Q-2- (2-Chaminothiaool-4-yl) acetamide J-3- (t- -methyl-1-pyrrolidinium) methyl-3-ce-35 fem-4-carboxylate ( ife).

A mixture of the compound from Example 4B (333 mg, 0.33 mmol) and N-methylpyrrolidine (60 mg, 0.7 mmol in (5 ml) was stirred for half an hour at room temperature and then evaporated in vacuo. The residue was washed ether and dissolved in 90% aqueous TFA. After standing for half an hour at room temperature, the mixture is concentrated under reduced pressure. To the concentrate, ether is added to separate the even vertical product, which is collected by filtration and dissolved in a small amount of methanol. The solution is chromatographed on an HP-2 column (40 ml). sc 30% aqueous -nm CHjOH followed by lyophilization gives 0.062 g of a mixture of isomers and 55 g ( "5: 1). The mixture was purified by HPLC (RP-18 Pihrosorb, 8x300 mm, 15% methanol) and the target isomer A

g-hell

501

j

ten ;

 . 15

- .20 . 25

, thirty

35

; 55 th

hell

736

(Ib) isolated as a pale yellow powder 4.9 mg (2.7%).

UV spectrum, cubic acid oxide (phosphorus buffer, pH 7), nm (t) 235 (15000); 258 (14,000).

NMR, JDjO, 1.43 (3N, t); 2.33 (4H, m); 3.10 (ЗН, s); 3.64 (4H, m); 4.3.6 (2H, q); 5.44 (1H, d); 5.95 (1H, d); 7.08 (1H, s).

EXAMPLE 5 7- (Z) -2- (2-npo-poxyimino) -2- (2-aminothiazol-4-sh1) acetamido} -3- (1-methyl-1-pyrrolidinium ) methyl 3-cephem-4-carboxylate (IiJ.

A. Diphenylmethyl ester of 3-chloromethyl-7- (Z) (2-propoxyimine) -2- (2-tritylaminothiazol-4-yl) acetamido-3-cephem-4-carboxylic acid.

A mixture of (E) -2- (2-propoxyimino) -2- - (2-triethylamino-thiaz ol-4-yl) acetic acid. (707 mg, 1.5 mmol) and phosphorus pentachloride (344 mg, 1.65 mmol) in dichloromethane (14 ml) are stirred at room temperature for 1 h and drunk in a solution of 7-amino-3-3-chloromethyl benzine ester -Z-cephem-4-carboxylic acid (677 mg, 1.5 mmol) and BSA (1.1 ml, 4.5 mmol) in dichloromethane (15 ml). The reaction mixture was stirred at room temperature for half an hour, diluted with ethyl acetate (200 ml) and water (3 X. 100 ml), dried over sodium sulfate and evaporated to give 1.4-g (100%) of the title compound.

: IR spectrum, (КВГГст: 3360; 3020, ЗОбО; 2960; 1785; 1725; 1680; 1520.. ": 1500; 1450; 1375; 1300;

125o; 1160; 1090; yubo; loio; 990;

840; 740; 700

UV spectrum, l EtOH, nm: 240 (24600); 260 S20700).

NMR, CDCl, 1.35 (6H, d, I 6 Hz); 3.50 (2H, s); 4.35 (2R; s); 4.58 (1H, m, 1 "6 Hz); 5.00 (1H, I - 4.5 Hz); 5.91 (1H, gg-, I 4.5 and 9 Hz; g for 1-4.5 Hz), 6.68 (1H, s), 6.88 (1H, s), 7.25 (25H , with).

B. Diphenylmethyl dt 3-yl-domethyl-7-L (Z) -2- (2-propoxymino) 2- - (2-tritylaminothiazol-4-ylacetamino-3-cephem-4-carboxylic acid ester).

A mixture of (Z) -2- (2-pro-oximemino--2- (2-triethylaminothiazol-4-ip) acetic acid (500 mg, 0.55 mmol). And iodide. Sodium (248 mg, 1, mcl) in acetone (10 ml) stirred at

room temperature for 50 min. After evaporation, the residue was dissolved in ethyl acetate (15 ml), washed successively with 10% aqueous sodium thiosulfate (10 ml), water (10 ml) and aqueous NaCl (10 ml), dried over sodium sulfate and evaporated to give 494 g ( 90%) of the indicated target compound.

 IR spectrum, (KBG), mV “s, article H 3360; 3040, - 3020; 2960-, 1785; 1720;

1080; 1boo; 1550; isoo; 1450; 1370; 1300; 123P; 1150; 1115; yuo; 990; 90o; 840; 730; 700

. UV spectrum,) EtOH, nm, (K): 240 (2A900); 260 (19400).

HMP, JCDC1 ppm: 1.30 (6H, dI 6 Hz), 3.37 and 3.70 (W each, d, I 16 Hz); 4.22 (2H, s), 4.55 OH, m, 1-6 Hz); 4.95 (W, g, I - 4.5 Hz); 5.83 (1H, g-g,, 5 9 Hz, g for D40) jL.6.66 (1H, s), 6.87 (1H. S); 7.25 (25 N, s).

C. 7- (Z) -2- (2-nponoKCHHMHHo) -2 -2 (2-aminothiazol-4-yl) acetamido-3- (1 st, ethyl-1-pyrrolidinium) methyl - I-3- cephem-4-carboxylate (Tc).

A mixture of the compound from Example 5B (545 mg, 0.55 mmol) and β-methyl pyrrolidine (70 mg, 0.82 mmol) in dichloromethane (10 ml) was stirred at room temperature for half an hour and diluted with ether (100ml) The resulting residue is collected by filtration. A solution of isadica in 90% TFA (4.5 ml) was stirred at room temperature for half an hour and evaporated in vacuo. The residue is triturated with ether to obtain 317 mg of crude product, which is chromatographed on an HP-20 column (50 mp), eluted with water (500 ml) and 3.0% CH, OH (500 ml). The eluate is concentrated in 30% and lyophilized.

Example 6. 7 G (X) -2-allyl-imino-2- (2-aminothiazol-4-yl) acetates (1-methyl-1-pyrrolidium) meth-3-cephem-4-carboxylate (7c1).

A. 7- (Z) -allyloxyimino-2- (2-trntidaminotol-sol-4-yl) acetamido-3-chloromethyl-3-cepheme-4-carboxylic acid 7- (Z) -allyl-amino-imidotinamide ester

In a suspension of benzyl ester-7-amino-3-chloromethyl-3-tsrfem-4-carbo

CH ,, pH

to obtain 109 mg of a mixture of isomers (

6/1), 100 mg of which was purified using HPLC (Likhrosorb RP-tS, 15% MeOH), to give 5 mg (3%) of the title compound by the order of 1C.

UV spectrum, “in 1K (PH 7, buffer), nor (f): 236 (15100), 252 (14600).

O, 1.42 (6H, d, 1-6 Hz); 2.33 (4H, s); 3.10 (ЗН, s);

ten

New acid (1.35 g, 3, mmol) in methylene chloride (20 ml) are added with B (1.1 ml, 4.5 mmol) and the mixture is stirred for-half a day at room temperature 15 to form a clear solution . A mixture of (2) -2-allyLok imino-2- (2-tritylaminothiazol-4-yl) hydrochloric acid (1.40 g, 3.0 mmol) and phosphorus pentachloride (690 mg, 20 3.3 mmol) in methylene chloride (20 ml is stirred for 15 minutes at room temperature and poured in one portion into a solution of trimethylsilylated compound V. 25 The mixture is stirred for 20 minutes at room temperature and diluted with acetate (200 ml), washed with aqueous sodium bicarbonate and water, dried and distilled off under reduced pressure. 30 The oil-like residue was purified on silica gel using column chromatography (Wako-he e, C-200, 30 g.) The column is blued with chloroform and the fractions containing the desired product are combined. Distillation under reduced pressure gives the desired product as an amorphous powder, yield 2.32 g (89%). M.P. 100-115 With (decomposition of the infrared spectrum (KBV), JMCIKC, cm:

40 3990; 1790; 1730; 1680; 1530; 138o;

1250; 1160; 1020.

NMR, CDCl,. 3.50 (2H,

a n); 4.32 (2n, s, 3-cHj.); 4.6-6.1

, (7H, m, CHjCHrCH, 6,7-H); 6; 70 (1H, 45 s, H thiazole); 6.90 (1H, s,); 7.1-7.6 (Zones, m, protons of phenyl).

Found,%: C 64, 13; 63.99; H 4.6 4.64; N 7.50, 7.30; S 6.85, 6.85; C1 7.55, 7.46.

35

50 C4gH NjOf S, C1-1 / 3 CHClj

. Calculated,%: C 64.05; H 4.45; N 7.73; S 7.03; C1 7.82.

B. Benzhydryl ester of 7- f (Z) - -2-allyloxyimino-2- (tritylaminoia3, 65 (4H, s); 3.83 and 4.23 (1H, each-55 sol) -4-yl) acetamido - 3-iodomethyl-3-, dy, d, Hz); 5.45 (1H, d, I - - cepheme-4-carboxylic acid. - 4.5 Hz) | 5.95 (1H, d, 5 Hz); A mixture of the compound from example 6L 7.05 (1H, s). . (2.30 g, 2.65 mmol) and sodium iodide

12501738

Example 6. 7 G (X) -2-allyloxy-imino-2- (2-aminothiazol-4-yl) acetam- (1-methyl-1-pyrrolidium) methyl-3-cephem-4-carboxylate (7c1).

A. 7- (Z) 2-Allyloxyimino-2- (2-trntidiaminothiazol-4-yl) acetamido-3-chloromethyl-3- Behydroxy ester 7- (Z). cepheme-4-carboxylic acid.

In a suspension of benzyl ester-7-α-amino-3-chloromethyl-3-tsrfem-4-carbo 10

New acid (1.35 g, 3, mmol) in methylene chloride (20 ml) are added with BSA (1.1 ml, 4.5 mmol) and the mixture is stirred for about 24 hours at room temperature 15 to form a clear solution. A mixture of (2) -2-allyLoxy-imino-2- (2-tritylaminothiazol-4-yl) acetic acid (1.40 g, 3.0 mmol) and phosphorus pentachloride (690 mg, 20.3 mmol ) in methylene chloride (20 ml) is stirred for 15 minutes at room temperature and poured in one portion into a solution of trimethylsilylated compound V. 25 The mixture is stirred for 20 minutes at room temperature and diluted with ethyl acetate (200 ml), washed aqueous sodium bicarbonate and water, dried and distilled off under reduced pressure. 30 The oil-like residue is purified-on silica gel using column chromatography (Waco-gel, C-200, 30 g). Column zyruyut chloroform and combine the fractions containing the target product. Distillation under reduced pressure gave the desired product as an amorphous powder, yield. 2.32. g (89%). T. pl. 100-115 C (decomposition). IR spectrum (KBG), JMCIKC, cm:

0 3990; 1790; 1730; 1680; 1530; 138o;

1250; 1160; 1020.

NMR, CDCl,. 3.50 (2H,

a n); 4.32 (2n, s, 3-cHj.); 4.6-6.1

(7H, m, CHjCHrCH, 6.7-H); 6; 70 (1H, 5 s, H thiazole); 6.90 (1H, s,); 7.1-7.6 (Zones, m, protons of phenyl).

Found,%: C 64, 13; 63.99; H 4.61, 4.64; N 7.50, 7.30; S 6.85, 6.85; C1 7.55, 7.46.

35

0 C4gH NjOf S, C1-1 / 3 CHClj

. Calculated,%: C 64.05; H 4.45; N 7.73; S 7.03; C1 7.82.

B. Benzhydryl ester 7- f (Z) - -2-allyloxyimino-2- (tritylaminoia-C2 g, 13.3 mmol) in acetone (15 ml) is stirred for 1 hour at a temperature of TJOHH and then evaporated under reduced pressure . The PSICT or oil-like residue in ethyl acetate (200 ml) was washed with sodium thiosulfate and water, evaporated under reduced pressure to give the compound as an amorphous powder, which was used in the next stage without further purification. Exit 2.52. g (99%).

C. 7- G (2) -2-apliLoximino 2- (2- -amino-thiazod-A-yl) acetamido-3- (1 - -methyl-1-pyrropidinyl) methyl 3 -3-cephem-4-carboxyl 1 (I ).

A mixture of the compound from Example 6B (478 mg, 0.5 mmol) and N-methylpyrrolidine (0.05 ml, 0.5 mmol) in methylene chloride (5 ml) was stirred for 20 minutes at room temperature and diluted with ether ( 50 ml) to precipitate a quaternary product (500 mg yield), a mixture of a quaternary product and TFA (2 ml) are allowed to stand at room temperature for one and a half hours and diluted with ether to precipitate a crude salt of TFA product (yield 265 mg), which chromatographic on a column of HP-20 (1.8x18 cm). The column was eluted with water and 30% aqueous methanol. The methanol eluate is evaporated under reduced pressure and the residue is lyophilized to give an amorphous powder (yield 124 mg), which contains a 35% pure product (17%) and the corresponding isrmer (83%). The mixture is purified by HPLC (Likhrosorb RP-18, 0.01 (pH 7); CHjOH "85:15). The eluate is acidified to pH 3. 40 is diluted with HCl and chromatographed on an HP-20 column (1.8 x 10 cm). The column is eluted with water and then with 30% aqueous methanol. The methanol eluate is evaporated under reduced pressure and; $ 4 residue is lyophilized to give.

of the indicated compound (t (i) as an amorphous powder (yield 13 mg, 5.1%). mp. 155 ° C (decomp.)

JK-spectrum, (KVg) Moiite cm: 50

3600-2800; 1770; 1670; 1610, 1530; 1200;

UV spectrum, (pH 7, buffer), nm (€): 235 (16600); 253 (15600) ..- 55

NMR; (PP, O, 2.1-2.5 (4H,. S, H pyrrolidine), 3.10 (3N, s, CHj); 3.4-3.8 (4H, m, pyrrolidine); 5.95

Uh.g; 4H, 2; H); 7.10 (1H, s, H thiazole),

Example 7. (2-aminothiazol-4-yl) - (g) 2-carboxyprop-2-oximemino-acetamidoJ-3- (1-methyl -1 -1-pyrrolidinium) methyl-3-cephem-4- - carboxylate (1 ").

A. Benzhydryl ester of 3-chloromethyl.-7-1 (g) -2- (2-tert-butoxycarbonyl-prop-2-oxyimino-2- (2-tritylamine-thiaol-4-yl) acetamido-3 cephem- - 4-carboxylic acid.

Method 1. A mixture of (Z) - -2 (2-tert-butoxycarbonylprop-2-hydroxyimino) -2-2-tritylaminothiazol-4-yl) acetic acid (1.94 g, 3.6 mmol), DDC ( 742 mg, 3.6 aviol) and H-hydroxybenzene triethanol (486 mg, 3.6 mmol) in tetragvdrofuran (T.HF) were stirred at room temperature for 45 minutes, during which time dicycine hydrochloride urea was separated. The latter is removed by filtration, and the filtrate is mixed with benzhydryloethyl 1m-7-amino-3-chloromethyl-3-cephem-4-carboxylic acid ester (1.5 g, 3.6 m / m). The mixture is stirred overnight at room temperature, and then evaporated in vacuo. The residual oil is dissolved in .CHClj (20 ml), washed with saturated aqueous NaHCO and saturated with aqueous NaCl, dried over MgSO4 and evaporated to dryness. The residue (3.9 g) was dissolved in n-hexane CHCl1 (1: 2) and passed through a silica gel column (40 g), using the same system, of solvents, the fractions containing the desired compound were evaporated in vacuo from semi-. 1.3 g- (39%) of the indicated compound melting at a temperature above (decomp.).

/ N-

IR spectrum, (KBG), m.v., .cm; 3990, - 1790; 1715; 1690.

UV spectrum, (EtOH), nm: 240 (E, 280); 265 (EC; 190).

NMR rfCDClj, 1.45 (ZN, s); 1.63 O 1.66 (6H, each s); 3.49 (2H, broad s); 4.34 (2H, s), 4.96 (1H, d, I - 4.5 Hz), 5.90 (1H, dd, I - 4.5 and 7.5 Hz); 6.66 (1H, s), 6.86 (1J, s); 7.0-7.5 - (25H, and); 8.23 (1H, d, I 7.5 Hz).

Me is wild 2. A solution of 7-amio-3-ormethyl-3-cephem-carboxylic acid bisyl ester (g, 4.49 mmol) in CHjCl

eleven

(A6.5 ml) of rbrsb are kneaded for AO minutes with obtaining a clear solution. To the solution was added a solution of acid chlorohydride, which is obtained from the compound of Sha (2.56 g, A, 49 mmol) and peij (1.12 g, 5.38 mmol) in methyl chloride (26 ml). The mixture was stirred at room temperature for half an hour, poured into cold water (100 ml) and extracted with ethyl acetate (3 x 50 ml). The combined ecotectum is washed with aqueous NaCl, dried and evaporated. The residual syrup (3 g) is chromatographed in a column of silica gel (100 g) eluted with 10, 1 with a mixture of goluol and ethyl acetate. The fractions containing the desired compound are combined and evaporated to give 2.84 g (65%) of this compound,

B. Benehydryl ester of 7- (Z) - -2 (2 tert-butoxycarbonylprop-1-hydroxyimino) -2- (2-tritylaminothiazol-4-yl) acetamide-J-3-iodomethyl-3-cefem-4 carboxylic acid. .

A mixture of the compound from Example 7A (500 mg, 0.53 mmol) and NaJ (240 mg, 1.6 mmol) in acetone (3 ml) was measured for 2 hours at room temperature and then evaporated in vacuo. To . GHjClji (20 ml) and ode (10 ml) are added. The organic layer is called 10% by volume. sodium thiosulfate solution (5 ml) and 1 ml of NaCl

1250173Г2

poured with ether (80 ml). Formed

the precipitate is collected by filtration and is triturated with ether / to afford 420 mg. quaternary product that

5 block with 90% TFA (4.2 ml) at room temperature for 1 Then the reaction mixture is evaporated to a dry residue. Add ether to the residue to obtain crude TFA salt

10 of compound .lo (245 mg, quantitative yield), which is a mixture of 1: 4 isomers l. Cheese product is purified by the method JHVY - (Likhrosorb RP-18: 4 x 3.00 m

 5 elution with 0.01 M ammonium phosphate buffer (pH 7.0), 10% content). The fraction containing the target is collected and evaporated to a large volume. Concentrate Argument

20 to a pH of about 2 by adding 1M HC1

and passed through a column (X 15 cm) to remove inorganic salts. HjO column flush

(1000 ml) and eluted with 30% CH, | OH 25 The eluate was evaporated and lyophilized to give 21 mg (10%) of the desired product (1e) as a colorless powder. T. pl. 160 C (decomp.). . IR spectrum, (КВг), с „(с 340 30 1775, 1610.

 UV spectrum pH 7 nm () (155500).

NMR, (Y 0,0, million

IMOKC phosphate buffer, 237 (15,700); 257

1.65 (6H, s);

p ml), dried over MgSO4; and evaporated 2,3 C4H, m); 3.09 (ZN, s), 3.7 (4H,

to a dry residue, taking into account 540 mg (50%) of the said compound as an amorphous powder melting at (decomp.).

40

m); 4.0 (2H, m); 5.44 (1H, d); I 4.8 Hz 5.94 (1H, d), 7.15 (1H, s

The compounds of formula I exhibit high antibacterial activity against gram-positive and gram-negative bacteria, are suitable for the treatment of bacterial infections of the stomach H1X as well as humans, can be used for the preparation of preparative forms intended for consumption by parenteral, in the usual way using well-known pharmaceutical, carriers and excipients and may be presented as single dosage forms or as multiple dosage forms. The compositions may be presented in type solutions, suspensions or emulsions in oily or aqueous diluents, and may contain conventional dispersing, suspending and stabilizing agents. Compositions M. may be presented in the form of dry. Cm

,-one

3350;

(s; 270);

IR spectrum (KVg) „1790, 1690.

UV spectrum, V EtOH: 240,265 (190).

NMR (JCDCl ,, 1.44 (9H, s); 1.65, (6H, s), 3.54 (W, 4.28 C2H, s); 4.98 (1H, d, I 4, 5 Hz; 5.85 (W, dd, I 4.5 and 7.5 Hz); 6.70 (1H, s); 6.90 (1H, s); 7.1-7, 5 (25H, m).

C. (2-aminothiaol-4-hyp) g (2) - -2- (2-carboxyprop-2-oxyimino) acetamido-3- (1-methyl-1-g1Hrrolidknium) methyl-3-cephem-- 4-carboxy am (1e).

The mixture of the iodomethyl derivative of Example 7B 038 mg, 0.51 mmol) and .Y-methylpyrrolide at (0.079 m, 0.076 mmol) a, (10.8 nl) are left to stand at room temperature for half an hour, and then dilute.

250173Г2

poured with ether (80 ml). Formed

the precipitate is collected by filtration and washed with ether / to obtain 420 mg of a quaternary product, which is

5 was blocked with 90% TFA (4.2 ml) at room temperature for 1 hour. Then the reaction mixture was evaporated to a dry residue. Ether is added to the residue to give crude TFA salt.

10 of compound .lo (245 mg, quantitative yield), which is a mixture of 1: 4 isomers l. The crude product is subjected to purification method JHVY - (Likhrosorb RP-18: 4 x 3.00 mm,

 5 elution with 0.01 M ammonium phosphate buffer (pH 7.0), 10% content). The fraction containing the target is collected and evaporated to a small volume. Concentrate made

20 to a pH of about 2 by adding 1M HC1

and passed through a column (2 x X 15 cm) to remove inorganic salts. HjO column flush

(1000 ml) and elute with 30% CH, | OH; 5 The eluate is evaporated and lyophilized to obtain 21 mg (10%) of the desired product (1e) as a colorless powder. T. pl. 160 C (decomp.). . IR spectrum, (КВг), с „(с 3400j 0 1775, 1610.

 UV spectrum pH 7 nm () (155500).

NMR, (Y 0,0, million

IMOKC phosphate buffer, 237 (15,700); 257

1.65 (6H, s);

2.3 C4H, m); 3.09 (ZN, s), 3.7 (4H,

2.3 C4H, m); 3.09 (ZN, s), 3.7 (4H,

m); 4.0 (2H, m); 5.44 (1H, d); I 4.8 Hz 5.94 (1H, d), 7.15 (1H, s).

The compounds of formula I exhibit high antibacterial activity. against gram-positive and gram-negative bacteria, suitable for the treatment of bacterial infections of animals Hb1X, as well as humans, can be used for the preparation of preparative forms intended for use parenterally, in the usual way: using known pharma. pharmaceuticals, carriers, and excipients, and may be presented as single dosage forms or as multiple dosage forms. -. The compositions may be presented in type solutions, suspensions or emulsions in oily or aqueous diluents, and may contain conventional dispersing, suspending and stabilizing agents. Compositions M. may be presented in the form of dry13

o Powder to be diluted before use, for example, sterile water that does not contain pyrogenic substances. The compounds of formula 1 may also be formulated as candlesticks using conventional bases for this purpose, such as cocoa butter or other glyceride. If necessary andredlagay. These compounds may be prescribed in combination with other antibiotics, such as leucicillins or other cephalosporins.

In the tea / tea 11rigotovlenn in the form of unit dosage form kompozii191I

Preferred B content is 50-1500 mg of the active ingredient Form1y || The dosage of adult treatment dosage is preferably 500-5000 mg / day, depending on the frequency and method of administration. In the case of intramuscular or intravenous administration to an adult, the total dose in porcine is usually 750-3000. mg / day in the form of separate dosages, although for some Moxet compounds, the daily dose in the case of

-

-coHH-r-rS i fi o dj ShgOsn

l

DOS

cefotaxime; connection for comparison

50173U

bacterium infections of the genus Pseudo-mpnas.

Preferred are compounds of formula 1 wherein R is methyl

5 or ethyl The most preferred compound with R is methyl. In the course of the initial evaluation of the proposed compounds, a shnimliy inhibitory concentration ((CC) for medianeny and two

10 compounds for comparison (cefotaxime and ((eft)) are determined by the method of double-dilution of agar using Mouller-Hinton agar for 32 strains of test organisms,.

13 groups divided into ioecTb.

The geometric mean MIC values defined in this activity are shown in Tables 2 and 5. Level 20 contents in KpoBii after intramuscular administration to mice (20 mg / kg) are given in Table. 3

Resistance (NICK / µg / ml) To one or several 2S testable compounds among 240 strains of Entebrocterlaceae in the Nuler-Quinton medium are shown in Table. four.

soon

I-conh-v

 livj LKaiy.V

-H-s

0

COC) ®

i c- jr-CH3: cooH

livj LKaiy

COC) ®

ceftaeidime, compound for comparison

HzN

; -coNH

0 “2

LcHrfO coo®

(one)

test compounds.

15

I All tested compounds are more active than cefotaxime in the otnoteniya groups (Q -) - II and (G -) - III tested organisms, with the most preferred compound I a being clearly more active. All tested compounds are more active than ceftazidime in relation to the (G +) - Ie and (G +) - 1 groups. test organisms, with the most preferred jo compound being clearly more active than ceftazidime in relation to all groups of test organisms, with the exception of (G -) - III, which are somewhat more sensitive to ceftazidime.

Absorption of the most preferred compound Jo and compounds for comparison (cefotaxime and ceftazodime) was determined in mice after a single intramuscular injection of the test compound (dissolved in 1M phosphate buffer; pH 7) at a dosage of 20 mg / kg. Blood samples were collected from orbital sinuses into capillary tubes treated with heparin and sown on Moller – Hinton medium using Morganella Morganii A 9695 as the test organism. Blood levels at different time intervals of the magnitude of the sub-periods. (ti /) and the area under the curves (AUC) are presented in Table. 3..

Tests were also conducted to expose organisms resistant to

5 o

5017316

preferably the compound of the formula Id, cefotaxime and ceftazide, the MIC values for these three compounds are. The scientific research institute in the ratio of 240 Entero-5 bacteriaceae strains whether the MJfK environment was equal to or greater than 8 dp of at least one of the tested individuals in the Mugol-Hinton medium was arbitrarily taken as evidence of the resistance of the organism. Of the 240. strains, 27 were resistant to at least one of the test compounds. From tab. 4 follows that 3 organisms are resistant to 15 compounds 1, 15 to ceftazidime and 18 to cefotaxime. Compound 1e is more active than cefotaxime in the otnoteniya group (G -) - II of the test organisms and much more active than cefotaxime in the 20 respect of the group (G -) - III of the test organisms (PS. Aeruginosa). It is more active than ceftazidime in relation to all groups of gram-positive test organisms, excluding (G -) - III. (PS. Aeruginosa), which are more sensitive to ceftazidnmu,

The test compound on the table. five,.

; -coNH-r-Ф Ф,

UWH-Tg W # X

 y-o (f l y% -VxJ (СНЗ

soon

Table 1

Note and e. (Sya -) - tv

(C) -T (G -) - lo

(5 strains), sensitive to ne scillin; (5 strains) sensitive to penicillin; (2.shtamma). К .1 Ш1гамм) and (2), sensitive to cefa Aotin ;; . (C -) - 1b - (3 strains) and (3 Strains), resistant to dafalothin;

Compound

1o, K-methyl (“) Cefotdxim (M Ceftazidnm (s)

Note: () - average value for 2 tests;

(b) - one trial;

(c) - average value for 3 tests.

J9

Escherichia coli Escherichia coli Klebsiella pneumoniae Enterobacter aerogenes Enterobacter aerogenes

Enterobacter cloacae

Enterobacter cloacae Enterobacter cloacae Enterobacter cloacae Citrobacter freundii

"

Citrobacter species Proteus vulgaris Morgdnella raorganii

Serratla marcescens Serratia marcescens Serratia marcescens Serratia marcescens Serratia marcescens

The total number of resistant strains.

1250173

20 Table L

27

15

18

n Rome e h an not. (G +) - Jlci - sensitive to penicillin

(5 strains); . () -Tb - sensitive to penicillin

  (5 strains); , (C) r1 sensitive to cefakotin

(2 strains), K1. (1 strain) and V, 2 strains); (G) -jl - resistant to cephalotin

(3 strains) and (3 strains); (G -) - II - (1 strain) ,. (2 strains) and

(2 strains); (O -) - lIl - (6 strains);

(o) average value for 5 ..sKcnepHMeHtoB.

Compiled by Z. Latypov. Presented A. Kozoriz Tehred N. Voikalo-Proofreader A. Zimokosov

Order 4341/60 Circulation 379. -Subscription

VNIILY 1 of the USSR State Committee

on affairs of inventions and O1gryty 113033, Moscow, Zh-35, Raushsk nab. d 4/5

Production and printing company, U Sgorod, ul. Project 4

Claims (1)

• METHOD OF OBTAINING ARBITRATION-: NEW CEPHALOSPORIN of the general formula where R is straight or branched. C ₄ -C ₄ -alkyl, allyl, 2-butenyl, 3-butenyl or 2-carboxypropyl, wherein the compound of the general formula: wherein R ₄ is straight or branched -C ₄ -alkyl, allyl, 2-bute. nyl, 3-butenyl or 2-carboxypropyl, in which the · 2-carboxy group is protected as an ester such as tert-butyl; In ₄ - a group that protects amino. a group such as tertiary; The ester forming group, such as diphenylmethyl or benzhydryl, is reacted with N-methylpyrrolidine in a solvent at room temperature, the protective groups are removed in the resulting product by acid hydrolysis and the desired product is isolated. Priority by feature. : 04.03.82 when R is straight or branched C ₄ -C ₄ alkyl, allyl, 2-butenyl or 3-butenyl; '' 03/12/82 at R - 2-carboxypropyl.