SU904533A3 - Method of preparing 7-(5-amino-5-carboxyvaleramido)-7-methoxy-3-(1-methyl-1h-tetrazol-5-yl)thiomethyl-delta-3-cephem-4-carboxylic acid or its alkaline metalsalts - Google Patents

Abstract

PURPOSE:7-Methoxycephalosporins of the formula II (R is heterocyclic thiols of the formula I, e.g. residue of 5-mercapto-1,3,4-thiadiazole-2-thioacetic acid of the formula III, or 5-mercapto-1-methyl-1H-tetrazol of the formurla IV).

Description

3 g The purpose of the invention is to obtain new antibiotics of a cephalosporic series and an expanding arsenal of effects on a living organism. This goal is achieved by a process for the preparation of compounds of the formula I, which consists in the fact that a stump is producing a 7-methoxycephalosporin-producing antibiotic microorganism belonging to the species Streptomyces in a culture medium containing nutrients, with the addition of 1-methyl-1H-tetrazole-5 -thylthiol of the formula, 1 or its salt, or its disulfide compound of the formula Thus, the target compound with a 1-methyl-1H-tetrazol-5-ylthiomethyl group in position 2 of the cephalosporin can be obtained directly in one stage of fermentation. Compound 1 has a high antimicrobial effect. The antimicrobial spectrum of compounds can be expanded or changed by replacing the side chain - the social group in the 7th position of the formula (1H- (CH2) for- 50 with another acyl group, for example, with the C-aminophenylacetyl group, with the C -carboxyphenylacetyl group, oC-sulfophenylacetyl group, o (α-oxyphenylacetyl group, pyridylthioacetyl group, thiadiazolylthioacetyl group, triazolyl acetyl group, cyanmethylthioacetyl group, trifluoroethylthioacetyl group, and t, p . Thus, the proposed compound is also suitable as an intermediate for the preparation of these derivatives with these acyl groups, for example, 7 (2-Cyanmethylthioacetamido-7-methoxy-3- {1-methyl-tetrazol-5-yl) -thiomethyl- A-cepheme - (- carboxylic acid and 7-methoxy-3- (1-methyltetrazol-5-ylthiomethyl) (- trifluoromethylthioacetamido) - -cephem- -carboxylic acid. An example of microorganisms useful in obtaining 7-methoxycephalosporin antibiotics, refer- with Streptomyces, can serve as a new strain of StreptomyCes Oganonensis VG 19 Z, previously isolated from the soil in Og o-Town, Khimibugun (Saitama prof., Japan), This strain was deposited at the Institute of Microbiological Industry, the Institute of Industrial Sciences and Technology of the Ministry of International Trade of Japan under 1 ° REKM-R 2725, as well as the American Type Culture Collection, 12301 Parklow Drive Gokville , Maryland 20852, USA, under the number of ATSS ZP67. Below are the mycological properties of this strain. 1. The morphological characteristics of the strain S. Oganonensis V-G 19 Z It grows in both natural and artificial media to form a well-branched substrate mycella, while the formation of aerial mycelium is insufficient and therefore the formation of spores is poor. Straight chains of spores are of type R (Rectus) or RF (Rectiflexibiles), each having 10-50 spores. Spores are elliptic, spherical or cylindrical in shape, size 0, - 0.60 x 0.55 - 0.90 microns. The surface of the dispute is smooth. Flagellary and sporangium spores are not observed. 2. The cultural characteristics of the strain S. Oganonensis V-G 19 Z are given in table 1. 3. Physiological properties shtamS .Oganonensis YG 19 Z «Formation Tyrosinase Negative Reduction of nitrate Positive Positive, Coagulation cream weak positive, weak Peptonization cream Polouhi Tel'nykh Hydrolysis of starch positive Liquefaction of gelatin 5E weak negative Decomposition of cellulose Hemolysis Positive Solubilization of calcium maleate A positive Utilization of carbon compounds S, oganonensis Y-6 19 Z. A source of uglerod Utilization Glucose + Arabinose + Sucrose Xylose "Inositol Manitol + Fructose + Ramnoza affinity) (the characteristic features of the strain Streptomyces oganonensis VG 19 Z are as follows: refers to the non-chromogenic strain Streptomyces; aerial mycelium is direct without mutant (type R or RF), spherical or elliptic spores, smooth surface; it gives a light yellow-gray to light yellow-brown culture on different media, air color; celee brown-white, yellow-linen 1 and yellow-gray; antibiotic VG 19 Z-DZ is produced, belonging to the goxycephalosporin group. When studying known strains with such properties, Streptomyces globlsporus is the closest. However, when compared with S.glob is porus, the strain VG 19 Z differs from it in the following paragraphs, shown in Table 2. From the data in the table it can be seen that the proposed strain is a new strain that differs from the known one. As Streptomyces ooanonensis related to the family of Streptomyces Streptomyces jumonjinensis, Streptomyces heteromorphus, Streptomyces panayensis and Strepto-. myces chartreusis. However, the proposed strains are not limited to the above mentioned strains; any strains belonging to the genus Streptomyces and producing 7 methoxycephalosporin can be used as well. antibiotics. 1-methyl-1H-tetrazol-5-ylthiol (II) can be used in the form of its inorganic salts, such as salts of alkali metals, alkaline earth metals, ammonium salts and the like, and salts of organic bases, such as salts of triethylamine, trietano amine, dicyclohexylamine, lysine, arginine, .histidine, basic water-soluble antibiotics, for example, kanaminine, alkaloid, basic protein, and so on, etc. Salts, having a high solubility in water, can be used selectively. In addition, the disulfide compound 1-methyl-1H-tetrazol-5-ylthiol (I) can be used. Cultivation is carried out according to the usual methods for all microorganisms, but it is better to conduct submerged cultivation in liquid culture. The foot is to use any culture medium containing nutrients for producing 7 methoxycephalosporin antibiotics strains belonging to the genus Streptomyces. As a carbon source, it can use glucose, sucrose, mannitol, glycerin, dextrin, starch, vegetable oils, etc., as a nitrogen source, meat extract, ceptone, glutemic flour, cotton seed flour, soy flour , peanut flour, fish meal, corn flour, dry yeast, yeast extract, ammonium sulfate, ammonium nitrate, urea and other organic and inorganic nitrogen sources. If necessary, metal salts can also be added to the culture medium, for example sulfates, nitrates, chlorides, carbonates, phosphates, etc., Na, K, MgSa, Zn, and Fe, as well as antibiotics or antifoaming agents. for example, methionine, cysteine, cystine, methyl oleate, lard, silicone oil, surfactants, etc. 1-Methyl-1H-tetrazol-5-yl-thiol (II) or its salt, or its disulfide compound (II) is usually added at a concentration of 0.1-5 mg / ml (preferably 0.5-2 mg / ml thiol). They can be added to the culture medium once before cultivation or in portions at the initial stage of cultivation. It is desirable to cultivate under aerobic conditions at a cultivation temperature of 18-35 ° (better than 30 ° C). In addition, the desired results are obtained with the pH of the culture medium (preferably 6-8). The duration of cultivation depends on the composition and temperature of the culture medium used, but usually it ranges from 3 to 10 days; thus, the target substance accumulates in the medium at the end of the cultivation. The target substance (1) can be isolated from the broth by the usual method used to isolate antibiotics from the culture broth mycelium: the target antibiotic is mainly contained in the culture broth, hence after the separation of the mycel by centrifuging the sludge by filtration, the target substances are extracted from the filtrate. Thus, the target substance is separated, isolated and purified by conventional methods characteristic of antibiotic production, for example, using the difference in solubility in the corresponding solvent, the difference in sorption agent for different sorbents, or the difference in distribution between the two liquids. These methods can be applied separately or in their respective combination, or repeatedly. The physical and chemical properties of the target compound are characterized by the following indicators: white powder shock, decomposes and becomes colorless at 160-170 °; readily soluble in; limited in methanol and poorly in other organic solvents; amphoteric substance with a positive inhydrin reaction; UV absorption spectrum when determined in phosphate buffer solution of 0.01 M at pH 6.4, the maximum absorption at 273 mmk; The IR spectrum, when determined as potassium bromide, is absorbed at 3413, 2920, 17b5, 8 1620, 1515 and .1390 cm-; The NMR spectrum defined in TMS as an external standard and heavy water gives the following signals: a value of 5 (ppm); 2.36 (H, multiplet), 2.96 (2H, multiplet), 3.98 (ZN, singlet), 38 (W, multiplet), 4.50 (ZN, singlet), 5.59 (1H, singlet), elemental analysis of the target substance in the purest form: C 37., and k, 2S, N 16.47, S 10.901; with hydrolysis 6 n. hydrochloric acid gives an oC-amino-pyridic acid, gives. 5-mercapto-1-methyl-1H-tetrazole by hydrolysis in methanol Dowex 50W (type H, trade name). From these data, it can be seen that the compound is a 7-methoxycephalosporin compound, since it gives signals at 3.98 ppm (OG, singlet, 7-OCHj) and 5.59 ppm (1H, singlet, 6 -CH) in the NMR spectrum, absorption in the IR spectrum at 1765 cm (cyclic lactam) and gives C-aminoadipic acid during acidic hydrolysis, in addition, the absorption goes at 4.50 ppm (ZN, singlet, tetrazole) M-methyl) in the NMR spectrum; in soft hydrolysis, 5 mercapto-1-methyl-1H-tetrazole is obtained. This compound has the above structure with a heterocyclic thiol. The results of the various chromatographic analyzes and paper electrophoresis for the target compound are given below. .The proposed Rf value of this compound in thin-layer chromatography using microcrystalline cellulose (Avicel SF, trade ° ° name), Target Compound 1 Cephalosporin C 7-methoxycephalosporin C Cefamycin C 0.26 0.26 YG 19 2-DZ YG 19 Z-A2 0 , 39 0.32 Apply solvent systems in the following ratio by volume: Isoprop, ol: butanol: acetic acid: 21: 3: 7: 9 water Butanol: acetic acid: water t: 1: 2 Butanol: acetic acid: water 6 : 1.5: 2.5 Control sample - cefamycin C is 7- (5-carboxivalamido) -3 carbamoyloxymethyl-7-methox1-cef M | carboxylic acid. V-G 19 Z-DZ and V-G 19 g-D2, used in comparative experiments, are new 7-methoxycephalospores compounds previously isolated from the culture fluid of Stre tomyces oganonensls. Below is the Rf value obtained by paper separation chromatography using Whatman No. 1 filter paper and a mixture of butanoacetic acid – water ratio in a ratio of 4: 1: 2 by volume. Target Compound I0.39 Cefalosporin C 0.36 7-methoxycephalos. Orin C 0.0 Cefamycin C 0.35 Y-6 19 Z-ДЗ 0.2 YG 19 Z-fl2 0.25 Then the compound is subjected to an anal Hitachi 635 High-Speed Liquid Chromatography Apparatus and get the following results: column - mm made of stainless steel; Hitachi 26 resin (cation exchange resin, trade name); solvent system 0.2 M citrate buffer solution (pH 3.6); feed rate — 0.5 ml / paper feed rate — I record, the instrument ero is 1.0 cm / min. Delay Time Target Compound Cephalosporin C 7 Methoxycephalosporin C Cefamycin C Y-G 19 Z-DZ Y-G 19 Z-D2 The analysis result for the above. The device under the following conditions is listed below. Column - | W Bondapak C (Walter Ltd.) mm; solvent system 10 - acetonitrile, a solution of acetic acid (pH 3i3) in the ratio 1: 9 by volume, - feed rate - 0.8 ml / min; feed rate of paper recorder 1, 0 cm / min. lag delay. (Target compound 1 min 52 s. The result obtained by high-voltage paper electrophoresis is the following: Whatman filter paper No. 1; solvent 10% acetic acid (pH 2.2); voltage 2 V / cm; duration 1 hour. Travel length Target compound 3, 3 cm. Cephalosporic 3.5 cm rin C 7-methoxy 3, cm phalosporin C 3.5 cm Cefamycin C 6,1 YG 19 Z-DZ 6,1 YG 19 2-D2 Cysteine 7.5 cm acid 1.2 cm of Glutathione in Table 3 shows the antimicrobial effect of the proposed compound together with cephalosporin C for comparison, Infusion agar disk method (pa 500 g / ml.) Numerical values indicate the diameter (mm) of the zone of inhibition. The proposed compound can be administered in various dosage forms as such or in combination with OTHER drugs. It can be administered orally, intramuscularly, intravenously, etc. capsules, tablets, powders, granules, solutions and suspensions. Various additives can be used to prepare the preparations, for example, mannitol, sucrose, glucose,. sterile distilled water, isotonic solution, vegetable oil, such as peanut oil, sesame oil. In addition, other ingredients can be added, for example, stabilizers, binders, antioxidants, preservatives, tabletting agents, suspending agents, viscosifying agents, fragrances, and the like. As salts of the compound, salts of inorganic or organic bases which are pharmacologically non-toxic are used. The dose of the drug depends mainly on the condition of the patient and the weight of the patient, as well as the method of administration, i.e. oral or parenteral route. Typically, 50 mg / kg is administered at a time several times a day.

Example 1. Preparation of 7- {5-amino-5-carboxivalamido) -3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-7-methoxy-D-cephem-carboxylic acid.

A culture medium containing 1 starch, glucose II, 1.5% soybean flour, 0.5 yeast extract, 0.1% potassium phosphate acid, 0.05% magnesium sulfate, and 0.3% sodium chloride is placed in Sakakuchi flasks 100 ml each and sterilized at 120 C for 20 min. Each flask is then inoculated with a strain of Streptomyces oganonensis V-G. 19 Z and cultured kQ h at 30 ° C. In addition, the above culture is placed in Sakaguchi flasks in 2 liter 00 ml each and after sterilization for 20 minutes each flask is inoculated up to 2-3% of the broth obtained above, and then cultured for 2k hours at 0 ° C to form - inoculum.

60 l of culture medium containing 7% starch, 2% gluten meal, 2% soy flour, 0.8% glycerol, 0.1 K1.1 SLOTS Casamino, 0.01% ferrous sulfate and 55 g sodium hydroxide are placed in two fermenters per 100 l together with 10 ml of Adecanol (trade name) as an antifoaming agent and after sterilization for 30 min at 12 (f 800 C 800 ml of inoculum are introduced into each fermenter, then culture is carried out for H h at 30 ° C. Then 1- methyl 5-mercapto-1H-tetrazole is dissolved in an aqueous solution of sodium hydroxide and the solution is sterilized under pressure and added to the stump iruemomu broth, placing it in the concentration of this solution was 0.05%, the mixture undertakings further cultured for 90 hours.

At the end of the cultivation, the broth is adjusted to pH 2.0 then mixed with Radiolite (trade name) with stirring. The mixture is filtered on a filter press and the filtrate is combined to give about 100 liters of filtrate. The filtrate is adjusted to pH 3.0 with an aqueous solution of sodium hydroxide, passed through a column with Amberlite CPD-2 (trade name) for 12 liters, and after washing the column with 30 liters of water, the column is eluted with 30 liters of 50% aqueous acetone solution. The eluate is evaporated to 5.5 liters and the concentrate is adjusted to pH 3.5 with a diluted solution and passed through a column with Amberlite 1RA-68 (chlorine type) (trade name) for 3 liters. The column is washed with 6 l of water and fractionated with an aqueous solution (pH 7.2) containing

5 1 M solution of sodium nitrate and 0.1 M solution of sodium acetate, get about 5 liters of solution containing the active antimicrobial substance. The solution is brought to pH 3.0, passed through a column with Amberlite HPD-2 (trade name) per 1 liter, washed with water and eluted with a 50% aqueous solution of acetone, get 00 ml of an aqueous solution,

5 containing an active antimicrobial agent. After lyophilization of the solution, about 5 g of a crude powder of compound II is obtained. The crude powder is subjected to chromatography.

0 on a column with 800 ml of Sephadex A-25 resin (acetic acid type) (trade name) with a small amount of 0.5 M ammonium bromide buffer solution in acetic acid to fractionate the active component. Anti-microbially active fractions are collected, passed through a column Amberlite HPD-2 (trade name) 500 ml, the column is washed with water and eluted with an aqueous solution of acetone. Fractions active against microbes are collected and evaporated to dryness in vacuo.

The precipitate formed is subjected to chromatography on a microcrystalline cellulose column (Avitsel) (torn name) filled with isopropanol: water (7: 3 by volume) mixed solvent, using the solvent system as described above. The obtained active antimicrobial fractions are collected, applied to a thin-layer plate of Avicell SF (trade name), treated with a butanol: acetic acid: water (6: 1.5: 2.5 by volume) solvent system and sprinkled onto the plate with 0.25% a solution of ninhydin-pi. oridine, then heated to wake up the staining. Then fractions with Rf 0.31 are collected. The fractions are evaporated in vacuo and dried, and the resulting residue is subjected to chromatography on a C with microcrystalline cellulose (Avice) using a mixture of solvents - iopropanol: butanol: acetic acid: water (volume ratio 21: 3: 7: 9). The obtained fractions possessing antimicrobial activity are subjected to thin layer chromatography on Avicel SF (trade mark) using a mixture of solvents of the same composition as above, and then, following the same operation as described above, the fractions with Rf 0 are collected. 39 and evaporate them in vacuo to dryness. 1 The residue thus obtained is chromatographed on a column with microcrystalline cellulose using a mixture of solvents butanol. Acetic acid: water (volume ratio 6: 1, 5) to purify the active component. The fraction thus purified is evaporated in a vacuum to dryness, dissolve in a small amount of distilled water and divided into columns with Sephadex G 10 (trade mark) using distilled water. Fractions possessing antimicrobial activity are collected and subjected to thin layer chromatography using a mixture of solvents butanol: acetic acid: water (volume ratio 6: 1.5: 2.5), as indicated above. Thereafter, fractions of Rf 0.31 are collected, concentrated, lyophilized, and about 60 mg of white 7- (5-amino-5-carboxyvalers to) -3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-7 are obtained -methoxy-D-cephem - (- carboxylic acid. Example 2. By the same method as used in example 1, in this example, using a solution of bis (1methyl-1H-tetrazol-5-yl) -disulfide prepared by dilution of the latter in water containing methanol and sterilized by filtration through a finely porous (Millipore) filter, instead of a solution of 1-methyl-5-mercapto-1H-tetrazole prepared dissolved By using it in water using an aqueous solution of sodium hydroxide, when it is sterilized under high pressure, 2b g of crude powder 9 of compound II is obtained, and by purifying it as in Example 2, about 37 mg of 7- (5-amino- 5-carboxivalamido) -3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-7-methoxy-L-cefem-C-carboxylic acid (compound I). Example 3. Dry matter capsules containing 120 mg of 7- (5-amino-5-carboxivalamido) -7-methoxy-3- (1-methyl-1H-tetrazol-5-yl) thiomethyl-A-cefem-carboxylic acid. Each capsule contains. 7- (5-A wno-5-carboxylic acid) -7-methoxy-3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-A-cepheme-4-carboxylic acid Lactose Magnesium stearate Capsule No. 3. 7- (5-amino-5-carboxivalamido) -7-methoxy-3- (1-methyl-1H-tetrazo-5-yl) -thiomethyl-D-cefem-carboxylic acid is ground to powder No. 60, then lactose and Magnesium stearate is sifted through a No. 60 fabric sieve onto the resulting powder, all ingredients are mixed for 10 minutes and the dry gelatin capsules are filled with this mixture. 3. Example k. Tablets containing 150 mg of 7- (5-amino-5-carboxy valeramido) -7-methoxy-3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-L-cephem-carboxylic acid. Tablets contain, mg: 7- (5-amino-5-carboxivalamido) -7-methoxy-3- (1-methyl-1H-tetrazol-5-yl) -thiomethyl-D-cephem - + - carboxylic acid 150 Acidic calcium phosphate IP115 Magnesium stearate 3 Lactose IP 39 The active ingredient is mixed with calcium phosphate and lactose, the mixture is granulated with 15 corn starch paste (mg) and sieved through a coarse screen, dried at tO С and sieved again through a K 16 screen, stearate is added Magnesium and rtpeccyt in tablets, approximately 8.5 mm in diameter. Example 5. Parenteral solution containing 500 mg of 7- (5-am no-5-cap6-oxivaleramido) -y-methoxy 3 {1-methyl-1I-tetrazol-5-yl) -thiol-A-cephem-carboxylic acid . Each vial contains 500 7- (5 amino-5-carboxivalamido) -7-methoxy-3- {1-methyl-1H-tetrazol-5-yl) -thiomethyl- (5: -cepheme-carnary acidic. Active the compound (50.0 g) was added to 150 ml of sterile water for injection, the resulting solution was adjusted to pH 8.0 with a dilute sodium hydroxide solution, and the solution was brought to 200 ml. This amount was divided into 100 ampoules, lyophilized and sealed ampoules. Example 6. Nutrient medium consisting of% starch, 1 ° i glucose, 1.5 soy flour, 0.5 yeast extract, 0.1 acid potassium phosphate and 0.05 sulphate rot, put into 500 ml Sakaguchi vessels in the amount of LLP ml and sterilized for 20 minutes at, then Streptomyces clavuligerus IF 013307 culture is sown into each container, and then cultured for 72 hours at a later time. aseptically transferred into 100 liters of Erlenmeyer flasks with a 500 ml capacity containing 50 ml of diluent consisting of 3 starch, 1.5% soy flour, 1.5 bolls of cotton, 0.5% sodium citrate, 0, Magnesium sulfate and 0.05 ferric sulfate hydrate, after which they are cultured with shaking at 2 7 ° C. After hours, a solution of 1-methyl-5-measure is prepared, then-1H-tetrazole according to the method described in Example 1, this solution is sterilized at high pressure and added to the nutrient medium, bringing its concentration to 0.6, after which the cultivation of the mixture continued in more. for 1bO h. After the end of the culture, the nutrient medium is filtered by pressure filtration and 2.5 liters of filtrate are obtained, the pH of the filtrate is adjusted to 2.0, washed with an equal amount of ethyl acetate, passed through a 50 ml Dowex 50 V column. (trade designation), the substance is washed in it and the substance is eluted from it with 125 ml of 0.2 M sodium phosphate solution (pH 4.6). The eluted solution is concentrated and separated on a microcrystalline cellulose column, from where the substance is eluted with a mixture of n- propanol; methanol: ode (7: 0.5: 2.5 by volume) to obtain in this way an aqueous solution containing the microbiologically active product. A similar procedure is also carried out using an n-butanol: acetic acid: water mixture (4: 1: 2 by volume). By lyophilizing the solution in this way, 29.5 mg of a powdered crude compound is obtained. The crude material is purified by liquid chromatography (Hitachi 2160 chromatograph, cationic ion exchange resin, 0.2 M citrate buffer solution at pH 3.6, flow rate 0.6 ml / min , the speed of movement of the diagram T cm / min). The microbiological active fraction, with an elution time of 6 minutes, is collected, concentrated, and then lyophilized, resulting in 9 mg of (5-amino-5-carboxyleramido) -7-methoxy-3- (1-methyl-1H-tetrazole). 1-yl) -thiomethyl-A-cephem-4-carboxylic acid having a white color. Similar operations are performed using Step, rochei 12908, Strep, limpanii IFO 13306 and Strep, viridochromgenes IFO cultures instead of Strep culture. clavuligerus tFO 13307 and get 28,23 and 25 mg of the crude product and 3 7 and 5 mg of the purified product respectively. Example 7. Each of the products obtained according to the experiments was dissolved in a small amount of distilled water and the pH of the solution was adjusted to a value of 6.0, sodium hydroxide was added to it, and the solution was lyophilized to give the corresponding sodium salts. The characteristics of the obtained salts, such as color reactions, IR, UV, MR spectra, refractive indices, chromatography on paper and liquid chromatography, solubility data are identical to those of the corresponding free acids.

17

90it533

18 Table 1

Good yellow sulfur to light brown

Nice light yellow

Good, light yellow to yellow brown

Good, light yellow brown

Nice, light yellow, brown

Moderate, cream

Good, light- Good, yellow-yellow-brown-gray to nevy brown BrownGood, olive- No forged-gray to dark olive

Good

Loffer serum medium

Not

Bad, gray

Slightly, light yellowish gray

Very light brownish gray

Light yellow brown

Very weak

Not

Brown-gray to dark yellowy gray

Yelto gray up

dark red

brown

Not

Not

19 O, S-0, 55 Spore size, µ Soluble pigment or glycerin-aspartic medium Utilization of rhinosis Negative Starch hydrolysis Strong Milk coagulation Positive Weak Milk Peptonization Milk production Cephalosporin antibiotics Positive

SOiiSSS

Claims (1)

20 Table 2 1.2-1, 8-2.0 or 0.9-1 L spherical Yellow to green yellow Positive Weak Negative Strong Negative 21EO BZ322 Invention formula - (1methyl-1H-tetrazol-5 yl) -thiomethyl1. The method of obtaining 7- (5 amino-5 of its salts with alkali metal for-carboxyvaleramido) -7-methoxy-3-muly I HOOC-CH- (CH2) 3-CONH-Uf -j V-lf in where M is a hydrogen atom or an alkaline or its salt, or its disulfide metal, a compound of formula I 11 characterized in that,; jf-- 7-tags-15 jj j producing | 4 a cephalosporin antibiotic microorganism belonging to type 1-.G Streptomyces, in culture medium containing nutrients, c. The method according to claim 1, about 1 t and h and adding 1-methyl-1H-tetrazole-5-20 o t and and with the fact that they use micl-ilthiol of the formula I: The organism of Streptomyces oganonensls Ij; jfY-G 19 2. jl i | Sources of information, NZ taken into account in examination I25 1. British patent Bf, CHjkl. C 2 A, pub. 1973. --cephem- -carboxylic acid or OCHz soy "