FI60211C - 7-AMINO-3- (TETRAZOLO (4,5-B) PYRIDAZIN-6-YLTHIOMETHYL) -3-CEFEM-4-CARBOXYL SYRA FOR ANALYZING SAUCE MELLAN PRODUCTS WITH FRAMSTATION OF THE ANTIBACTERIAL 3-ACYL ACID - Google Patents

Description

GaSCT ΓβΊ MULTIPLE * PUBLICATION

IB] (11) utlAgcni NGSSKRIFT 6021 1 «SB C 10 IS 1901

Patent medJelat ^ v ^ (51) Kv.ik.3 / Int.a.3 c 07 D 501/18 ENGLISH — FI N LAN D (21) Puenttlhik.mu.-Ptnttniöknlni 79253 * + (22) H> k «MltpUvl - An * 6knlng * dtg 15.08.79 (23) AlkupUvi —Glttifh * udas 27.08.73 (41) Tullut lulkMctl - Bllvlt offantllf 15 · 08.79

Patent * and the National Board of Registration .. „« .ι, · ....., ..,. .....

'. . , (44) NlhtlylksIptiKHi and moonLjalsalsun pvm. - οη no ο.

The patent and registration are disclosed and the publication discloses -3X · u °. oi (32) (33) (31) Claim claimed — Bajird priorltet 30.08.72 06.09.72 USA 281 + 792, 286793 (71) Bristol-ftyers Company, 3 * + 5 Park Avenue, New York, NY, USA (US) (72) Takayuki Naito, Tokyo, Jun Okumura, Yokohama, Hajime Kamachi, Yokohama, Japan-Japan (JP) (7 * +) Oy Kolster Ab (5 * +) Antibacterial 3-thiolated 7 ~ acylaminocephalosporanic acid derivatives 7-amino-3- (tetrazolo [1,5-b] pyridazin-6-ylthiomethyl) -3-cephem-* -carboxylic acid 7-amino-3- (tetrazolo / 1,5-b7pyridazine) used as an intermediate in the preparation of 6-ylthiomethyl) -3-cephem-1-carboxylic acid for the preparation of other products with antibacterial activity against 3-thiolated 7-acylaminocephalosporan compounds (62) Separated from application 2670/73 (patent 57111) -Avdelad frän ansökan 57111)

The invention relates to an intermediate of the formula ch2nh2

(f \ - (CH?) -C-Ntf-J- "T" SvS

C00H

wherein n is 7-amino used in the preparation of the antibacterial 7- (o-aminomethylphenylacetamido and propionamido) -3-tetrazolo [*, 5-b7-pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid according to 1 or 2, 6021 -3- (Tetrazolo [1,5-b] pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid »of the formula H2N_- Ί 1 in

C00H

7- (o-Aminomethylphenylacetamido and propionamido) -3-tetrazolo [5-b] pyridazin-6-ylthiomethyl) -3-cephem-U-carboxylic acids have been found to be valuable bactericidal agents, a dietary supplement in animal feed, and a mastitis inflammatory agent. as therapeutic agents in poultry and other animals, including man, in the treatment of infectious diseases caused by many gram-positive and gram-negative bacteria.

There is a wealth of literature dealing with cefa-losporin antibiotics. The main prior art data in this field are presented below: 7-Phenylacetamidocephalosporanic acid, also referred to as the N-phenylacetyl derivative of 7-AGA, cephaloram, PACA, and apparently also phenazporine are well known in the scientific literature. Publications concerning the preparation and / or properties of this compound with or without substituents on the benzene ring and of the corresponding compounds in which the 3-acetoxymethyl group has been replaced by a methyl, hydroxymethyl and / or pyridinium group are as follows: A '·· 3 60211

Chauvette, R.R., et al. "Chemistry of Cephalosporin Antibiotics II. Preparation of a New Class of Antibiotics and the Relation of Structure To Activity", Journal of the American Chemical Society, 84 3401-4302 (1962).

Chauvette, R.R., et al. "Structure-Activity of Relationship: ':; Among 7-Acylamidocephalosporanic Acids", Antimicrobial Agents and Chemotherapy -1962, 687-694.

Cocker, J.D. et al., "Cephalosporanic Acids, Part II. Displacement of the Acetoxy Group by Nucleophiles", Journal of the Chemical Society, 5015-5031 (1965).

Cocker, J.D. et al., "Cephalosporanic Acids. Part IV. 7-Acylamido-Ceph-2-em-4-carboxylic Acids", Journal of the Chemical Society, 1142-1151 (1966).

Culp, H.W., et al., "Metabolism and Absorption of the 7- (Phenyl-acetamido-1-C '' Li) -cephalosporanic Acid", Antimicrobial Agents and Chemotherapy - 1963, 243-246.

Jago, M., "Antibacterial Activity of Some Derivatives of 7-Amirio-cephalosporanic Acid Against Staphyloccus Auereus and Synergism Between These and Other Antibiotics," Britt. J. Pharmacol., 22-23 (1964).

Loder, B., et al. , "The Cephalosporin C Nucleus (7-aminocephal.o-sporanic Acid) and Some of Its Derivatives," Biochemical Journal, 79, 408-416 (1961).

Nishida, M., et al. "Studies on Microbial Degradation of Cephalosporin 'C Derivatives II", The Journal of Antibiotics, 21, 375-478 (1968).

Nishida, M., et al., "Studies of Microbial Degradation of Cephalosporin C Derivatives I.", The Journal of Antibiotics, 21, 165-169 (1968).

Spencer, J.L., et al., "Chemistry of Cephalosporin Antibiotics VIII. Synthesis and Structure-Activity Relationships of Cephaloridine Analogues", Antimicrobial Agents and Chemotherapy - 1966, 573-580. Stedman, R.J., et al., "7-aminodesacetocycephalosporanic Acid and its Derivatives", J. Med. Chem., 7 (1), 117-119 (1964).

4,60211

Sullivan, H.R., et al., "Metabolism of Oral Cephalothin and Rexlated Cephalosporins in the Rat," Biochemical Journal, 102, 976-982 (1967).

Vymola, F., et al., "The Classification and Characteristics of Cephalosporin Antibiotics I. Systematic Study of the Quantitative Sensitivity of Some Pathogenic Microorganisms to Cephaloridine", Journal of Hygiene, Epidemiology of Microbiology and Immunology, 10, 180-189 (1966) .

Many other 7-acyl derivatives of 7-aminocephalosporanic acid have been reported in the patent literature, including 7- (4- (α-aminoalkyl)? 9-acetylido) cephalosporanic acids (U.S. Patent No. 3,582,241).

7 - ((p-aminophenyl) acetamido) cephalosporanic acid (U.S. Patent No. 3,422,100), 7- (halophenylthioacetamido) cephalosporanic acids (U.S. Patent No. 3 * 335,136). and an almost unlimited number of variations of the compounds included in the general formula (and often not otherwise described) disclosed in patents such as Dutch Patent 69/02013 (Farmdoc 39 * 172). 7- (p-aminophenylacetamido) cephalosporanic acid is disclosed in U.S. Patent No. 3 * 422,103 as well as the corresponding N-trityl derivative (see also Japanese Patent No. 2712/67) (Farmdoc 25,406).

U.S. Patent No. 3 * 219 * 662 claims compounds having the structure R-CH 2 -CCO-ACA, wherein R is phenyl, nitrophenyl (especially para-nitro), chlorophenyl, alkylphenyl and alkoxyphenyl. a group, and the corresponding phenoxy and substituted compounds, and all the corresponding compounds in which the 3-acetoxymethyl group is replaced by a 3-pyridinium methyl group. A broader group of such compounds, including a series in which R is a phenylthio group, and also compounds in which R is a benzyl group (i.e., 7- (P-phenylpropionamido) cephalosporanic acid), an alkoxybenzyl, alkanoyloxybenzyl, aminobenzyl group, etc. are disclosed for use at least in general. as starting materials in English Patent Nos. 1,012,943 »and 1,153,421 (Farmdoc 23,984), see also English Patent No. 1,001,478 and U.S. Patent No. 3,280,118. Other 7-phenylacetamidocephalosporanic acids having substituents on the benzene ring, such as hydroxy and amino groups, are disclosed as starting materials in British Patent Nos. 1,082,943 and 1,082,962.

U.S. Patent No. 3,341,531 describes 7- (o-, m- and p-carboxyamidomethylphenylacetamido) cephalosporanic acids and their betaines. A number of 7- (halo-, dihalo-, nitro-, and halonitrophenylacetamido) ceflosporan acids are mentioned as starting materials for the reaction with certain nucleophiles in U.S. Patent No. 3,431,259 (Farmdoc 27,715) · Other 7- (phenylacetamido) cephalosporanic acids the benzene ring has various substituents are disclosed in Japanese Patent No. 2712/67 (Farmdoc 25,406), Japanese Patent No. 26105/69 (Farmdoc 40.86θ), English Patent No. 1,178,471 (Farmdoc 27,715, see Dutch Patent 67 / OO906) and Japanese Patent No. 25785/69 (Farmdoc 40,847) ·

Replacement of the 3-acetoxy group of a cephalosporin with various heterocyclic thiols is described in a) South African patent 70/2290 (see also Dutch patent 70/05519 (Farmdoc 80,188 R)), in which the side chains were e.g.

7-α-aminophenylacetamido groups and typical heterocyclic thiols included 2-methyl-1,5,4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5-thiol; and b) in U.S. Patent No. 3,516,997, wherein the 7-chain side chains had a structure such as R 1 - (alk) m -CO-NH- and R 1 -S- (alk) m -CO-NH-, wherein R 1 was one of many aromatic heterocyclic groups, and numerous heterocyclic thiols at the 3 ”position were, e.g., 1-methyltetrazole-5-thiol and 2-methyl-1,3,4-thiadiazole-5-thiol, and c) U.S. Patent 3,563,983.

Various cephalosporins, including cephalosporin C by chance, have been reacted with nucleophilic, aromatic mercaptans to produce compounds having the structure

/OF

Acyl-NH-CH-CHOHo

III

O = C - N2, C-CH2-S-Ar

COOH

In U.S. Patent No. 3,278,531, Ar is a phenyl group or a particular substituted phenyl group or a particular aromatic heterocyclic ring designated, e.g., in column 5. · Similar nucleophiles, e.g., 2-mercaptopyrinidines, are disclosed in U.S. Patent Nos. 3,261,832; 3,479,350 and 3 * 5.02,665 and in British Patent 1,101,422, all published in the name of Claxo. A similar representation can be found in the English patent 1,109,525 published in the name of Ciba, e.g.

6021 1e from the position "h" denoting the group R ^. Additional nucleophiles of this type were disclosed by Fujisawa in Belgian Patent 714 * 518 (Farmdoc 35,307; Dutch Patent 6θ / θ6ΐ29 and South African Patent 2695/68), Canadian Patent 818,501 (Farmdoc 38,845) »British Patent 67/148) and British Patent 1,187,323 (Farmdoc U.S. Pat. No. 3,516,997 (Farmdoc 34,328; Dutch Pat. 1969, American Society for Microbiology, Bethesda, Maryland, pp. 236-243 and J. Antibiotics (Japan) 23 (3), 131-148 (ΐ97θ).

More recently, the replacement of the 3-acetoxy group of a cephalosporin with various heterocyclic thiols has been disclosed in U.S. Patent 3,563,903 and Dutch Patent 70/05519 (Farmdoc 80,188 R) in which the side chains were e.g. 7-α-aminophenylacetamido groups and typical heterocyclic thiol cyclic , 4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5-thiol; the latter corresponds to U.S. Patent 3,641,021, published February 8, 1972 on the basis of an application filed April 18, 1969. «Other similar disclosures can be found in U.S. Patent No. 3,563,983» Belgian Patent No. 771,189 (Farmdoc 12,817 T), Japanese Patent 72/05550 (Farmdoc 12,921 T) and Japanese Patent 72/0551 (Farmdoc 12,922 T).

Various cephalosporins with structure

Acyl-NH-CH-Cli CH

I 1 I

0 = C-N C-CH 2 -C 5 alkyl »

COOH

where acyl represents various side chains including an α-aminophenylacetyl group is described in some of the above patents and in Glaxo's Belgian Patent 734,532 (Farmdoc 41,619) and Belgian Patent 734,533 (Farmdoc 41,620) and U.S. Patent 3,668,203 ·

Cephalosporins having the structure 7 60211 ^ s \

Acyl-NH-CH-CH CEj,

I I I

0 «C — N C-CH9-X

g COOH 0 wherein X comprises -S-C- and -S-C- are disclosed in some of the above patents and U.S. Patent 3,239,515; 3,239,516; 3,245,435 # 3,258,461; 3,431,259 and 3,431,259 and 3,446,803.

Related publications in the scientific literature are J. Med. Chem. 8, 174-181 (1965) and J. Chem. Soc. (London) 1595-1605 (1965) »5015-5031 (1965) and 1959-1963 (1967).

6-Mercaptotetrazolo (4,5-b) pyridazine was described by B. Stanovnik et al., J. Org. Chem. 35 (4), 1138-1141 (1970) as compound 8 (R = H) under the name 6-mercapto-tetratolo [1,5-b] pyridazine.

There are various presentations in the literature on methanesulfonates of antibiotics. Accordingly, Belgian Patent 742,728 (Farmdoc 41466 R) describes ampicillin monomethanesulfonate. U.S. Patents 3,268,508 and 3,295,246 describe the conversion of one or more of kanamycin from four free amino acids to a methanesulfonate derivative. Methanesulfonate sodium betrazine is disclosed in U.S. Patent 3,205,137 and for neomycin see J. Antibiotics (Japan) 12, 114-115 (1959). The injectable antibiotic keelistomethane (marketed by Warner-Chilcott as "Coly-Mycin") is the sodium salt of colistin methanesulfonate; The Merck Index cites Koyama et al., Japan 4898 (1957) as an example. A detailed review of coliethomethane is provided on pages 315-320 in U.S. Pat. Dispensatory 26th Edition, J.B. Lippincott Company, Philadelphia, Fenns. It is described as the pentasodium salt of the Penta (methanesulfonic acid) derivative of Colist's Preparation A on pages 621-622 in U.S.P. XVII. U.S. Patent 2,599,950 discloses in Example VIII a "polymyxin-formaldehyde-sodium bisulfite compound." See also English Patents 874,028; 902,992 and 896,774 and Japanese Patent 15948/61.

8 60211

The final products of formula I ch2nh2 / = \ "/ N ^ \ ^ - (cH2) nC-» H-CH — CH CH, »'-' jfi -N C-CH -S - </) - ^ Ν * Ne ' 2 \ = /

COOH

prepared by acylation according to formula II / S \

HpN ~ 9H-CH \ h_ n- ^ ^ SL

COOH

compound with an acid of formula III CH, »™ m

/ \ ^ (CH 2) n C H °°

wherein B is an amino protecting group and n is 1 or 2 to give a compound of formula ch2nhb C \ 2 / "% s ff (CH,) n ~ C-NH-CH» ", V ^»

COOH

wherein B and n are the same as above, and then the amino-protecting group B is removed to give a compound of formula I.

These reactions are described in more detail in FI patent publication 57111.

5,60211

The compound of formula II can be prepared by reacting 7- & amino-cephalosporanic acid or a salt thereof with a thiol of formula li

HS - ^ _ N

or a salt thereof, preferably a sodium or potassium salt.

Replacement of an ester group of a 7-aminocephalosporanic acid, e.g. a 3-acetoxy group with a thiol group, is a well known reaction and can be carried out in aqueous solution in the presence of a dilute base such as sodium bicarbonate. The replacement is preferably performed at a temperature between about 50-100 ° C.

In the following examples, all temperatures are in degrees Celsius. 7-Amino-cephalosporanic acid is abbreviated as 7-ACA; -ACA- represents a radical having the structure / s \ -NH-CH-CH CH0 I 1 1 / N \ f ^ C'C "2 '

COOH O

n and thus the 7-ACA compound can be represented by the formula H-ACA-O-C-CH 2.

Example 1 7-Amino-3- (tetrazolo- (H.5-b) pyridazin-6-ylthiomethyl) -3-cephemic carboxylic acid

/ N

H2Ni- <N —N. I

ί-γ ^ - ^ - 0 · 000 ^ + hs '\ = ^) ==' N --V

co2h 7-ACA · 13 c ° 2h ''

II

10 6021 1 i) To a hot solution (50-60 ° C) of 9 * 56 g (0.062 mol) of tetrat Solo / ΐ *, 5-b7pyridazine-6-thiol (13) and 10l2 g (0.12l mol) sodium bicarbonate in 300 ml of water, 16.86 g (0.062 mol) of 7-ACA were carefully added and the mixture was kept at 80-85 ° for 30 min. About 7 g of sodium bicarbonate was added to the reaction mixture to dissolve the insoluble matter. The solution was treated with activated carbon, filtered and the filtrate was acidified to pH 5 with dilute hydrogen chloride. The precipitate was collected by filtration, washed with water, air dried and finally in vacuo over P 2 O 2 to give 11.17 g (61%) of the title compound, m.p. 218-250 ° C (decomposes).

ii) A stirred solution of 16.8 g (0.11 moles of tetrazolo [4,5-b] pyridazine-6-thiol) and 18.18 g (0.22 moles) of NaHCO3 per liter of 0.1- M phosphate buffer (pH 6.1) was heated at 50 ° C and 30 g (0.11 moles) of 7-ACA was added portionwise to the solution. After heating at 2.5 ° C for 2.5 hours, the mixture still contained insoluble matter. The reaction mixture was cooled to room temperature and the precipitated title compound was collected by filtration, washed thoroughly with 200 ml of water and air dried.

Additional product was obtained from the filtrate and washings by adding dilute HCl to pH 5 · Total yield 32.9 g (83%). Sp. 215-250 ° C (decomposes).

IR: Y 1800, 1615, 1538, 1360 cm -1.

* max 'UV: γ 1 * NaHCO 237 nm (t 19500), 275 nm (£ 12000), 310 nm (sh) (δ 5700).

Δ max NMR: SD 2 O + K 2 CO 2, 3.35 (1H, d, 18 Hz, 2-H), 3.76, (1H, d, 18 Hz, 2-H), ppm 1.00 ( 1H, d, 10 Hz, 3-CH 2), 1.18 (1H, d, 10 Hz, 3-CH 2), 1.93 (1H, d, 1 Hz, 6-H), 5.32 (1H, d, 1 Hz, 7-H), 7.16 (1H, d, 10 Hz, pyridazine-H), 8.18 (1H, d, 10 Hz, pyridazine-H).

Calculated from the formula: C 12 H 11 N 2 O 2 Sg: C, 39, M; H, 3.03; N, 26.83; S, 17.55 Experimental: C, 39.19; H, 2.71; N, 26.81; S, 17.35 ·

The following examples illustrate the preparation of 7- (o-aminomethylphenylacetamido and propionamido) -3- (tetrazoloyl, 5-bypyridazin-6-ylthiomethyl) -3-cephem-1-carboxylic acids from the intermediate of formula (II).

11 6021 1

Example 2

+ t-BuOCON ^ _ ^ j ^^ - ^ \ nH-BOC 2,4-DNP, DCC

B

Jl6 N.

/ ^ TJ

N-N i,.

H-ACA-S- ^ V ^ N (lT>

r ^ V ^ NH-BOC _ \ = J

UjLvCO 0 -G ~ NO 2 __> NOg

XL

NH-BOC N

CFvCOOH

U JL, 00-NH --- N- / f 5 - *

C02H

18 OC - r-rs ί N- ^ r Ä / c ^ -S * (j = "

0 CO2H

20 12 6021 1 t-Butoxycarbonyl azide (15) To a cooled solution of 100 g (0.76 moles) of t-butyl carbazate in 87 g of glacial acetic acid and 120 ml of water was added dropwise a solution of 60 g (0 , 85 moles) of sodium nitrite in 50 ml of water for 1 + 0 min. while maintaining the temperature at 10-15 ° C. At the end of the addition, stirring was continued for another 30 min. at the same temperature. To the mixture was added 100 ml of water, and the separated oil was extracted with five 100 ml portions of methylene chloride. The combined organic extracts were washed with 100 ml of 10% sodium bicarbonate solution and 100 ml of water successively and dried over anhydrous sodium sulfate. The methylene chloride was removed under reduced pressure with a water bath maintained at 1 * 0-1 I + 5 ° C. The remaining azide was distilled and recovered at ii5 ° C and 20 mm Hg. It weighed 92.7 (8U%).

o- (t-butoxycarbonylaminomethyl) phenylacetic acid (16)

To a solution of 70 g (0.35 moles) of o-aminomethylphenylacetic acid hydrochloride (U) and 11 g (1.15 moles) of triethylamine (TEA) in 1 * 00 ml of water was added dropwise a solution of 6U g (0 , 1 * 5 moles) of t-butoxycarbonylthiazide (15) in 300 ml of tetrahydrofuran (THF) with stirring at 0 ° C. At the end of the addition, the temperature was allowed to rise to room temperature and stirring was continued for 20 hours. Tetrahydrofuran was distilled off at less than 1 * 0 ° C, and the aqueous solution was washed with 200 ml of ether, layered with 200 ml of ethyl acetate, and acidified with dilute hydrochloric acid to pH 3 under cooling to 0 ° C. The organic layer was separated and the aqueous layer was extracted with four 200 mL portions of ethyl acetate. The combined ethyl acetate solution was washed with 200 ml of water, dried over anhydrous sodium sulfate and concentrated in vacuo. The concentrate was treated with 500 mL of n-hexane to give 87.9 g (95%) of product as 16 colorless needles that melted at 11-116 ° C.

2, L-Dinitrophenyl-α-butoxycarbonylaminomethylphenyl acetate (17) Dicyclohexylcarbodiimide (17.72 g, 0.086 mol) (DCC) was added in one portion to a mixture of o- (t-butoxycarbonylaminomethyl) phenylacetic acid, (16) 73 g, 0.086 mol) and 2,2 + -dinitrophenol (15.82 g, 0.086 mol) (2.1 * -DNP) in 250 ml of THF. The reaction mixture was stirred for 2 hours at room temperature. The precipitated dicyclohexylurea was filtered off and washed with 100 ml of THF. The filtrate and washings were combined and concentrated under reduced pressure below 50 ° C to give a viscous yellow oil which was triturated in n-hexane (150 mL) to give the product 17 as yellow needles. Yield 3M (91%). Sp. 76-77 ° C.

33U0, 1785, 1685, 1610, 15U0, 1530, 1500, 13 ^ 0 cm -1.

ZD.8LX

Calculating from the formula ^ 20 ^ 21 ^ 3 ^ 8: ^ 5 * 68; H, U, 91; N, 9.7

Experimental: C, 55.70; H, 5.05; N, 9.93.

13 6021 1 7- (α-Butoxycarbonylaminomethyl-phenylacetamido) -3- (tetrazolo- (1,5-b) pyridazin-6-ylthiomethyl) -3-cephem-1-carboxylic acid (18) To a solution of 20.26 g ( 0.017 mol) of product 17 in 150 ml of tetrahydrofuran, a solution of 11.10 g (0.039 mol) of the compound of formula (II) and 19.19 g (0.19 mol) of TEA in 150 ml was added. in a $ 50 aqueous solution of tetrahydrofuran at 0-5 ° C. The reaction mixture was stirred for 18 hours and concentrated under reduced pressure to remove tetrahydrofuran below 30 ° C. The aqueous concentrate was washed with two 200 mL portions of ether, the acids were taken up in pH 2 with dilute hydrochloric acid and extracted with five 200 mL portions of ether, acidified to pH 2 with dilute hydrochloric acid and extracted with five 200 mL portions of ethyl acetate. The combined extracts were washed with two 100 ml portions of water, dried over anhydrous sodium sulfate, treated with activated carbon and filtered. Evaporation of the solvent gave a pale yellow oil which was triturated with ether to give 13.89 g (58%) of product 18.

Sp. 166-173 ° C (decomposes).

fCRr »—Ί ffi: ^ max 1770 '1710» 1690 »1535' 1370. 1255, 1170 cm.

Analysis calculated for .H 2 O: C, 19.51; H, 1.79; N, 17.77, S, 10.17.

Experimental: C i + 9.58; 19.65; H, U »15i 1.59; N, 17, Ui; 17.66; S, 10.1.

7- (o-Aminomethylphenylacetamido) -3- (tetrazolo (1,5-b) -pyridazin-6-ylthiomethyl) -3-cephem-U-carboxylic acid (20) To 20 ml of trifluoroacetic acid was added in one portion 13.80 g ( 0.022 mol) of product 18 and the mixture was stirred for 15 min. 0.10 ° C. To the reaction mixture was added 300 ml of ether to give the product 20 as its trifluoroacetate, which was collected by filtration and washed with ether. The trifluoroacetate was dissolved in 20 ml of water and the pH was adjusted to 5 with ammonium hydroxide to give the zwitterion as a gum, which was separated by decantation and triturated with water. The solid product was collected by filtration, washed with 20 ml of water and 200 ml of acetonitrile successively and dried in vacuo over phosphorus pentoxide to give 5.10 g (U5%) of product 20, which was fairly pure but amorphous.

Recrystallization of product 20 - an amorphous powder (3.50 g) was dissolved in 100 ml of a 50% aqueous solution of tetrahydrofuran with heating to 60-70 ° C with vigorous stirring. The solution was treated with a small amount of activated carbon. The filtrate was cooled, the walls of the vessel were scraped with a glass rod and the solution was allowed to stand overnight in the refrigerator to give 2.27 g of product as 20 fine needles melting at 190-193 ° C (decomposes).

Analysis: Calculated for C 18 H 18 N 2 O 3 Sg.ljSHgO: C, 16.76; H, 1.30; N, 20.77; S, 11.89.

Experimental: C, 17.18; 1 ?, 37; H, 1.08; 3.88; N, 20.93; 20.23; S, 12.03.

* - · · 'lii 6021 1

Example 3 CH-CCCH-CCCEt ^ H3 r ^ V ^ KHg 9_ ^ K | p ^^ HO = CH-COGEt k ^ V / COCH C ^ Hj-ONa ^ ✓ "'k ^ CCCNa i. 32« - <' (f »^ CICCCEt H-AC / rS-P) —H r ^^ r ^^ KH-OCH-COOEt

-> -V—> kA ^ CC-NH ^ K_M / K

O-i ^ Nv LCH2 S

^ co2h ^ KCCCH ^ CC ^ T-r \ N_N ^ co2h 20.

Sodium o- (1-ethoxycarbonyl-1-propen-2-ylaminomethyl) phenylacetate (19).

To an alcoholic solution of sodium ethoxide (sodium metal 5.75 g (0.25 g atom) and absolute ethanol 500 ml) was added 41.26 g (0.25 mol) of o-aninomethylphenylacetic acid (obtained by neutralizing the hydrochloride with 4 aqueous ammonia) and 52.5 g (0.25 mol). moles) of ethyl acetoacetate sequentially. The mixture was refluxed for 6 hours and treated with activated carbon and filtered through diatomaceous earth ("Dicalite"). The filter pad was washed with 200 mL of hot ethanol. The combined filtrate and washings were evaporated to near dryness and cooled to 0 ° C to give product 19 as colorless needles which were collected by filtration, washed with 200 ml of ethanol and dried in vacuo over PoOc. Solid product 19 weighed 44.51 g · Additional product 19 was obtained by concentrating the mother liquor. Total catch 56.51 g ($ 79).

Sp. 251-252 ° C (decomposes).

i5 6021 1 7- (o-Aminomethylphenylacetamido) -3- (tetrazolo (4,5-b) pyridazin-16-ylthiomethyl) -3-cephem-4-carboxylic acid (20)

Ethyl formate (6.87 g, 0.0063 mol) was added in one portion to a stirred suspension of 17.44 g (0.057 mol) of sodium O- (1-ethoxycarbonyl-1-propen-2-ylaminomethyl) phenylacetate (19) 200 per ml of dry THF containing 1 ml of Ν, Ν-dimethylbenzylamine at -15 ° C. Stirring was stopped and a cooled solution of 20.80 g (0.057 moles) of product II and 9.60 g (0.095 moles) of triethylamine in 2CO ml of a $ 50 aqueous solution of THF was slowly added along the wall. The mixture was stirred vigorously for 30 min. At 0-15 ° C, treated with activated carbon and filtered through "Dicalite" diatomaceous earth. The layer was washed with 50 mL of a 50 mL aqueous solution of THF containing 8 mL of triethylamine. Formic acid (3 mL) was added to a combined solution of the filtrate and washings to precipitate unreacted 7-ACA (2.5 g), which was filtered off. The filtrate was mixed with 200 ml of ether and then with 15 ml of formic acid. The mixture was stirred for 10-15 min. at room temperature and the resulting precipitate was collected by filtration, washed with 100 ml of ether and 500 ml of water successively and dried in vacuo over PgSO 4 to give 23.05 g ($ 79) of product 20, m.p. 180-186 ° C (decomposes).

Recrystallization of product 20 - The amorphous product described above (12.93 g) was dissolved in 1.2 L of THF at 50-60 ° C with vigorous stirring, treated with 5 g of activated carbon and filtered. The filtrate was seeded and stored in the refrigerator overnight to give 7.60 g of product as 20 fine needles. Sp. 186-189 ° C (decomposes).

Example ** A suspension of 7- (o-aminomethylphenyl azotamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid ionic form (0.361 g) in 3 ml of methanol is cooled with ice and treat with a few drops of concentrated hydrochloric acid until a clear solution is obtained. 7- (o-Aminomethylphenylacetamido) -3- (tetrazolo (4,5) pyrazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid hydrochloride precipitates as a light brown solid upon addition of ether and is collected by filtration and dried. stupid helpers ·

Example 5 To a stirred suspension of the zwitterionic form of the zwitterionic form of 3- (o-aminomethylphenylacetamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid (0.361 g) is added an aqueous solution of 1 N sodium hydroxide at room temperature. until a clear solution (pH 10.8) is obtained. This solution is freeze-dried immediately to give the crude solid sodium 7- (o-aminomethylphenylacetamido) -3- (tetratolo [4,5-b] pyridazin-6-ylthiomethyl) -3-cephem-4. carboxylate instead.

Method for preparing dimethanesulfonate Example 6 OT2NH2

COOK

? 2H5 + HOCH ^ SOjNa CH ^ (CH2) 5CH-C00Na (30 i »S EH in acetone)

Heat in water 2 · Treat with charcoal and filter 3 · Add filtrate to anhydrous alcohol.

V

CH 2 N (CH 3 SO 3) g

f \ "/“ \ / H

Solid C VCH2-C-NH-CH-CH CH2 N — j </ _ \ c · l COONa

HgO | - EHA, 1T 60 21 1

Procedure:

Place 2.0 moles (about 1025 g on the anhydrous basis) of 7- (o-aminomethylphenylacetamido) -J- (tetrazolo- (4,5-b) -pyridazin-6-ylthiomethyl) -3-cephem-4- carboxylic acid, 540 g of sodium formaldehyde bisulfite (4.03 moles), 5000 ml of water and 2700 ml (4 to 87 moles) of $ 50 SEH (sodium 2-ethylhexanoate) dissolved in acetone in a suitable tank and heat the mixture with stirring to 40-45 ° C. The mixture dissolves in about 10 minutes to a yellow solution.

After heating for 15 minutes, add 50 g of decolorizing carbon ("Darko KB") to the solution and stir for another 15 minutes. 40-45 ° C esa.

Filter through diatomaceous earth (Dicalite) after maintaining the reaction mixture at 40-45 ° C for a total of 50 min.

Wash the carbon cake with 2000 ml of $ 50 aqueous ethanol. Combine the extracts, adjust the temperature to 25 ° C, and add the solution at 25 ° C to 112 liters of rapidly stirred $ 100 ethyl alcohol. A fine, white, amorphous precipitate is formed which is sodium di (7- (o-aminomethylphenylacetamido) -3- (tetratolo (4,5-b) pyrazin-6-ylthiomethyl) -5-cephem-4-carboxylate di ( sodium methanesulfonate).

Stir the suspension for about 10 min. and then filter and wash the cake with 15 1 of $ 100 ethyl alcohol.

Dry the cake at 50-55 ° C in an oven with air circulation for about 2 hours and then under vacuum at 4-6 mm for 24 hours.

The yield is about 1200-1400 g of amorphous, white, solid sodium 7- (o-aminomethylphenylacetamido) -5- (tetrazolo (4,5-6) pyridazin-6-ylthiomethyl) -5-cephem-4-carboxylate di (sodium methanesulfonate). The product usually contains a few percent water and possibly a small amount of ethanol.

This product may also be called sodium-7- (oN, N-bis (sodium sulfomethyl) aminomethylphenylacetamido) -5- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -5-cephem- 4-karbokeylaatti.

An example?

The following slurry is prepared: 2.19 g of sodium formaldehyde bisulfite (2 equivalents) 3.5 g of 7- (o-aminomethylphenylacetamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -5-cephem-4- carboxylic acid zwitterion (100-200 mesh).

25 ml of water (volume can be varied).

14 ml of 50% SEH isopropanol solution 18 6021 1

An almost finished solution is obtained after rapid stirring for about 0.5 hours at 24 ° C. The temperature of the mixture is rapidly raised to 40-43 ° C. This is maintained for about 2 minutes and then rapidly cooled to 20-23 ° C.

The solution is filtered to remove small insolubles (total dissolution time should not exceed 2 hours).

A solution with a pH of 7 * 3 is added for 5 min. to 600 ml of very rapidly stirred absolute ethanol (other alcohols such as anhydrous isopropanol may be used). An amorphous precipitate of sodium 7- (o-aminomethylphenylacetamido) -3- (tetratololo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate di (sodium methanesulfonate) is formed. The mixture is stirred for 3 min. The precipitate is collected by filtration, washed with 60 ml of ethanol (or isopropanol) and dried in vacuo at 50 ° C for 24 hours. The catch is about 4 # 3 g.

The product is soluble in water at a pH of about 7 in an amount of at least 200 mg / ml. Such a solution is stable for at least 2 hours at room temperature; dilute solutions are stable for longer. The product shows the same bactericidal properties as its predecessor zwitterion and is fully biologically active whether or not hydrolyzed back to the zwitterion.

! Example 8

Sodium 7- (N, N-bi8- (sodium sulfomethyl) aminomethylphenylacetamido) -3- (tetrazolo (4,3-b) pyridin-6-ylthiomethyl) -3-cephem-4-carboxylate A. Preparation using hydroxymethanesulfonate

A mixture of 1 g (1.95 mmol) of 7- (o-aminomethylphenylacetamido-3- (tetrazolo (4,5'-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid , 1.52 g (10 mmol) of sodium hydroxymethanesulfonate monohydrate, 6 ml (6 mmol) of a 1 M solution of SEH in ethyl acetate, 10 ml of isopropanol and 10 ml of water were stirred at room temperature for 3.5 hours, and the resulting solution was treated with 1 g of activated carbon and it was poured with stirring into 300 ml of absolute ethanol and the mixture was stirred at room temperature for 30 minutes to give a crystalline product which was collected by filtration, washed with three 30 ml portions of absolute ethanol and dried over PgO 2 at 45-52 ° C for 1 mm. 1.5 h to give 1.51 g of sodium 7- (oN, N-bis (sodium sulfomethyl) aminomethylphenylacetamido) -3- (tetrazolo- (4,5 “b) pyridazin-6-ylthiomethyl) -3-cephem -4-carboxylate, which was readily soluble in water (> 1 g / ml), mp> 270 ° C.

j 'is 6021 1 IE: rrar 1760, 1660-1620, I605, 1540, 1400, 1200, 1040 cm -1.

IDftX

TTV: λmax 242 nm (ε 18,000), 271 nm (ε 11,600 sh), 312 nm (ε 5,100, sh), max NMR: δ D2 3.34 (1H, d, 1Θ Hz, S-CH2), δ , 76 (1H, d, 1 Hz, S-CH 2), 3.85 ppm (2H, s, CO-CH 2 - / {), 4.04 (4H, N-CH 2 - & 3-CH 2 -S), 4.30 (4H, s, N-CH 2 SO 2 Na), 4.92 (1H, d, 4.5, Hz, 6-H), 5.42 (1H, d, 4.5, Hz, 7- H), 7.05-7.30 (4H, phenyl-H), 7.40, (1H, d, 10 Hz, pyridazine-H), 8.11 (1H, d, 10 Hz, pyridazine- B).

B. To be prepared using formalin and sodium bisulfite a) To a solution of 1 mL (10 mmol) of formalin and 1 g of sodium bisulfite in 10 mL of water was added sequentially 1.026 g (2 mmol) of 7- (o-aminomethylphenylacetamido) - 3- (Tetrazolo (4,5-6) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid, 6 ml of 1-M SEH solution and 10 ml of isopropanol. The mixture was stirred for 2.5 hours at room temperature and poured into 300 ml of ethanol. The obtained sodium 7- (oN, N-bis (sodium sulfosethyl) aminomethylphenylacetamido) -3- (tetrazolo (4,5'L) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate was collected by filtration. , washed with three 50 ml portions of ethanol and dried in vacuo. Hall 1, 6l g.

b) To a mixture of 1.026 g (2 mmol) of 7- (o-aminoacetylphenylacetamido) -3- (tetrazolo (4,5-6) pyrazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid, 6 ml of a 1 M solution of SEH in ethyl acetate, 10 ml of isopropanol and 10 ml of water, 1 ml (10 mmol) of 30 l of formalin were added. The mixture was stirred for 2 hours at room temperature to give a clear solution of a small amount of oily precipitate. After 1 g of sodium bisulfite was added to the solution, it was stirred for another 2 hours, during which time an oily precipitate dissolved in the solution. The reaction mixture was poured into 300 ethanol with vigorous stirring to give 1.61 g of sodium 7- (oN, N-bis (sodium sulfomethyl) aminomethyl-phenylacetamido) -3- (tetrazolo (4,5-6) pyridazin-6-ylthiomethyl). ) -3-cephem-4-carboxylate, which was collected by filtration, was washed with three 50 ml portions of ethanol and dried in vacuo.

Example 9 A, 7- (B- (o-t-butocarbonylaminomethylphenyl) propionamido) -3- (tetratolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid 20 ° 6 0 21 1

N, N-dicyclohexylcarbodiimide (0.41 g, 2 mmol) was added to a mixture of 8- (α-butoxycarbonylaminomethylphenyl) propionic acid (0.56 g, 2 mmol) and 2,4-dinitrophenyl (0.37 St 2 mmol) 5 ml * sea THF and the mixture was stirred for 1 hour at room temperature. The precipitated urea was filtered off. To the filtrate was added a solution of 7-β®1ηο-3- (tetrazolo (4 ', 5') pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid (0.75 g, 2 mmol) and triethylamine (0 , 61 g, 8 mmol) in 10 mL of 30 aqueous THP and the mixture was stirred for 18 h at room temperature. The reaction mixture was washed with ether (2 x 20 mL) and the aqueous layer was acidified to pH 2 with dilute HCl and extracted with ethyl acetate (3 x 50 ml). The combined extracts were washed with water (30 ml), treated with activated carbon, dried over anhydrous Na 2 SO 4 and evaporated to an oil under reduced pressure. The oil was triturated in 50 mL of ether to give 7- (3- (α-butoxycarbonylaminomethylphenyl) propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid · As a colorless solid which was collected by filtration and dried in vacuo over P 2 O 2.

Yield 0.33 (26 #). Sp. 110-120 ° C (decomposes).

m: 1780, 1710, 1690, 1530, 1370, 1250 cm -1.

UV: ^ 1 NaHCO 3 ^ ^ ^ 1? 400) 270 nm (sh) (ε 12,500), 310 nm (sh) maX (e 5000).

NMR: bp δ ° D 6 1.37 (9H, s, t-Bu-H), 4.96 (1H, d, 4 Hz, 6-R), 5.55 (1H, dd, 4 & 8 Hz , 7-H), 6.99 (4H, e, phenyl-H), 7.57 (1H, d, 10 Hz, pyridazine-H), 8.38 (1H, d, 10 Hz, pyridazine-H) , 8.69 (1H, d, 8 Hz, CONH).

Analysis calculated by the formula: C 20 H 18 NQO 2 S 2 * 1/2 HgO: C, 51.02; H, 4.91; N, 17.6 Experimental: C, 51.74; H, 4.83; N, 17.88.

B. 7- (3 '(o-Aminomethylphenyl) propionamido) -3- (tetrathibolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid

Trifluoroacetic acid (0.5 ml) and 7- (P- (o-tert-butoxycarbonylaminoacetylenyl) propionamido) -3- (tetratololo (4,5-b) Shirts g in-6-ylthiomethyl) - 3-Cephem-4-carboxylic acid (0.28 g, 0.46 mmol) was stirred with cooling to 0 ° C and stirred for 30 min. Ether (50 ml) was added to the mixture to give 7- (3- (o-aminomethylphenyl) propionamido) -3- (tetrazol (4,5-b) -pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid trifluoro acetate, which was separated by decantation, washed with ether, dissolved in water (1 mL) and the pH was adjusted to 6 with dilute NH 4 OH. The product 7- (8- (o-aminoacetylphenyl) propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiooxyethyl) -3-cephem-4-carboxylic acid was collected by filtration and dried. was evacuated in vacuo over PgO 2. Yield 0.10 g ($ 42), m.p. 190-197 ° C (decomposes).

IB: γ13 * 1760, 1665, 1600, 1535, cm -1.

ΟΙαλ

Analysis calculated from ®20: 49 * 33} H, 4 * 33} N, 20.92} S, 12.18.

Experimental: C, 49.04} H, 4.26} N, 20.17} S, 11.96.

Example 10 A. Sodium p- (o- (1-ethocycarbonyl-1-propen-2-ylaminomethyl) phenyl) propionate

To an alcoholic solution of sodium ethoxide (sodium metal 5 * 75 g (0.25 g atom) and absolute ethanol 500 ml) is added 0.25 mol of P- (o-aminomethylphenyl) propionic acid (obtained by neutralizing its hydrochloride with aqueous ammonia) and 32.5 g (0.25 moles) of ethyl acetoacetate in succession. The mixture is refluxed for 6 hours and treated with activated carbon and filtered through diatomaceous earth ("Dicalite"). The filter pad is washed with 200 ml of hot ethanol. The combined filtrate and washings are evaporated to near dryness and cooled to 0 ° C to give sodium O- (o- (1-ethoxycarbonyl-1-propen-2-ylaminomethyl) phenyl) propionate as colorless needles which are collected by filtration, washed with 200 ml of ethanol and dried in vacuo over P 2 O 3. Additional product is obtained by concentrating the mother liquor. Total catch about 50 g ·) 3. 7- (p- (o - aminomethylphenyl) propionamido) -3- (tetrazolo (4 ', 5 ~ b) "^ Ψ · 1 - ... - m, m. Im ι I - -.1111 1 I" "1 1 -1 1 1 - (.... ....... - "Pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid

Ethyl chloroformate (6.87 g, 0.0063 mol) is added in one portion to a stirred suspension of 0.057 mol of sodium B- (o- (1-ethoxycarbonyl-1-propen-2-ylaminomethyl) phenyl) propionate in 200 ml: in dry THF containing 1 ml of Ν, Ν-dimethylbenzylamine at -15 ° C. Stirring is stopped and a cooled solution of 20.80 g (0.057 mol) of 7-amino-3- (tetrazolo (4 * 5 ~ 6) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid and 9, 60 g (0.095 mol) of triethylamine in 200 ml of 50 aqueous THF are slowly added along the wall. The mixture is stirred vigorously for J min. 0-15 ° C, treated with activated carbon and filtered through Dicalite diatomaceous earth. The layer is washed with 50 ml of a 50% aqueous solution of THF containing 8 ml of triethylamine. Formic acid (3 ml) is added to the filtrate. and a combined solution of washing solutions to precipitate unreacted 7-ACA (2.5 g), which is filtered off, 200 ml of ether 22 6 0 21 1 and then 15 ml of formic acid are stirred in. The mixture is stirred for 10-15 minutes at room temperature and the precipitate obtained is collected. by filtration, washed with 10 ° C ether and 500 ml water successively and dried in vacuo over P 2 O 2 to give 7- (O- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (4 * 5- b) -pyridazin-6-ylthiomethyl) -5-cephem-4-carboxylic acid about 20 g.

Recrystallization of 7- (3- (o-aminomethylphenyl) propionamido) -5- (tetrazolo (4 ', 5') pyridin-6-ylthiomethyl) -3-cephem-4-carboxylic acid - The amorphous product described above (13 g) is dissolved in 1 .2 l of a 50% aqueous solution of CFP at 50-60 ° C with vigorous stirring, treated with 5 g of activated carbon and filtered. The filtrate is seeded and stored in the refrigerator overnight to give about 8 g of product as fine needles.

Example 11 7- (O- (o-Aminomethylphenyl) propionamido) -3- (tetrazolo (4 - 5 - *) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid a suspension of the zwitterionic form (0.361 g) in 3 ml of methanol is cooled with ice and treated with a few drops of concentrated hydrochloric acid until a clear solution is obtained. 7- (O- (o-Aninomethylphenyl) propionamido) -3- (tetrazolo (4,5-6) pyridazin-6-ylthio · methyl) -3-cephem-4-carboxylic acid hydrochloride precipitates as a light brown solid upon addition of ether and it is recovered and dried in vacuo over PgO 2.

Example 12 To a stirred suspension of the zwitterionic form of the zwitterionic form of 3- (O- (o-aminomethylphenyl) propionamido) -3- (tetrazolazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid (0.361 g) is added aqueous 1-N sodium hydroxide solution at room temperature. until a clear solution (pH 10.8) is obtained. This solution is freeze-dried immediately to give crude solid sodium 7- (O- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (4 ', 5-6) pyridazin-6-yl · thiomethyl) -3-hefem -4-carboxylate.

23 6021 1

Methods for preparing dimethanesulfonate Example 13 ch2nh2 j— f .sv Λ AcHjj— «a-c-im-cH — jm ch2» _ £ f / ~~ Nxc / ~ CHa'Sn (_ /

COOH

fsH5 + HOCHgSO ^ Na -f- CH ^ CHg) jCH-COONa · (30/6 S EH in acetone) 1. Heat in water 2. Treat with charcoal and filter 2. Add filtrate to anhydrous alcohol Ψ CH2N (CH2SO3Ma) 2 8 / s \ Λ c * COONa h20 __L_ eha.

* 2k 6 0 21 1

Procedure:

Set 2.0 moles (calculated as about 10 52 8 anhydrous) of 7- (O- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3- cefem-4-carboxylic acid, 540 g of sodium formaldehyde bisulfite (4 → 03 moles), 3000 ml of water and 200 ml (4.67 moles) of a $ 30 solution of SEH (sodium 2-ethylhexanoate) in acetone in a suitable container and heat the mixture with stirring. 40-45 ° C. The mixture dissolves to a yellow solution in about 10 minutes. After heating for 15 minutes, add 50 g of decolorizing carbon ("Darco KB *") to the solution and stir for another 15 minutes at 40-45 ° C. Filter through diatomaceous earth ("Dicalite") after holding the reaction mixture. At 40-45 ° C for a total of 20 min.

Wash the carbon cake with 2000 ml of a $ 50 aqueous ethanol solution. Combine the filtrates, adjust the temperature to 25 ° C, and add the solution at 25 ° C to 112 L of rapidly stirred $ 100 ethyl alcohol. A fine white amorphous precipitate is formed which is sodium 7- (P- (o-aminomethylphenyl) -propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4- carboxylate di- (sodium methanesulfonate).

Stir the suspension for about 10 min. and then filter and wash the cake with 15 1 of $ 100 ethyl alcohol.

Dry the cake at 50-55 ° C in a convection oven for about 2 hours and then under vacuum at 4-6 mm for 24 hours.

The yield is about 1200-1400 g of an amorphous, white, solid sodium 7- (B- (o-aminomethylphenyl) propionamido) -3- (tetrazolo- (4,5,6) pyridazin-6-ylthiomethyl) -3-cofem- 4-carboxylate di (sodium methanesulfonate).

The product usually contains a few percent water and possibly a small amount of ethanol.

This product is also known as sodium 7- (P- (oH, N-bis- (sodium sulfomotyl) aminomethylphenyl) propionamido) -3- (tetrazolo (4,5-fc) -pyridazin-6-ylthiomethyl) -3-cephem- 4-carboxylate.

Example 1 ^

Prepare the following slurry: 2.19 8 sodium formaldehyde bisulfite (2 equivalents) 3,5 8 7- (P- (o-aminomethylphenylpropionamido) -3- (tetrazolo (4,5-b) pyridathin-6-ylthiomethyl) -3 -cephem-4-carboxylic acid zwitterion (100-200 mesh) 25 ml water (volume can be varied) 14 ml 30 $ SEH isopropanol solution.

An almost finished solution is obtained with rapid stirring for about 0.5 hours at 25 ° C. The temperature of the mixture is rapidly raised to 40-45 ° C. This is maintained for about 2 minutes and then rapidly cooled to 20-23 ° C.

The solution is filtered to remove small insolubles (total dissolution time should not exceed two hours).

A solution with a pH of 7 «3» is added for 5 min. to 600 ml of very rapidly stirred absolute ethanol (other alcohols such as anhydrous iropropanol may be used). Forms amorphous sodium 7- (p- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate di (sodium methanesulfonate) sediment. The mixture is stirred for 3 min. The precipitate is collected by filtration, washed with 60 ml of ethanol (or isopropanol) and dried in vacuo at 30 ° C for 24 hours.

The catch is about 4 g.

The product is soluble in water at a pH of about 7 in an amount of at least 200 mg / ml. Such a solution is stable for at least 2 hours at room temperature; dilute solutions are stable for longer. The product exhibits the same bactericidal properties as its preceding zwitterion and is biologically fully active whether or not hydrolyzed back to the zwitterion.

Example

Sodium 7- (P- (oN.N-bis (sodium sulfomethyl) aminomethylphenyl) propionamido) -3- (tetrazolo (4.5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate A. Preparation using hydroxymethanesulfonate A mixture of 1.95 mmol of 7- (P- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate -acid, 1.52 g (10 mmol) of sodium hydroxymethanesulfonate monohydrate, 6 ml (6 mmol) of a 1 M solution of SEH in ethyl acetate, 10 ml of isopropanol and 10 ml of water are stirred at room temperature for 3.5 hours. The resulting solution is treated with 1 g of activated carbon and poured with stirring into 300 ml of absolute ethanol, and the mixture is stirred at room temperature for 30 minutes. to give a crystalline product which is collected by filtration, washed with three 50 ml portions of absolute ethanol and dried over PgO 4 at 45-52 ° C at 1 mm for 20 hours to give about 1.5 g of sodium 7- (p- (oN, N-bis (sodium sulfomethyl) aminomethylphenyl) propionamido) -3- (tetrasolo- (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylate, which is readily soluble in water ( > 1 g / ml).

26 6021 1 B. Preparation using formalin and sodium bisulphite (a) To a solution of 1 ml (10 mmol) of 30 # formalin and 1 g of sodium bisulphite in 10 ml of water is added successively 2 mmol of 7- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (1 +, 5-b) pyridazin-6-ylthiomethyl) -3-cephem-1-carboxylic acid, 6 ml of 1-M SEH solution and 10 ml of isopropanol. The mixture is stirred for 2.5 hours at room temperature and poured into 300 ml of ethanol. The resulting sodium 7 - (/ ®- (oN, N-bis (sodium sulfomethyl) aminomethylphenyl) propionamido) -3- (tetrazolo (U, 5-b) pyridazin-6-ylthiomethyl) -3-cephem-U- the carboxylate is collected by filtration, washed with three 50 ml portions of ethanol and dried in vacuo. Catch about 1.5 g.

b) To a mixture of 2 mmol of 7- (S- (o-aminomethylphenyl) propionamido) -3- (tetrazolo (1+, 5-b) pyridazin-6-ylthiomethyl) -3-cephem-1-carboxylic acid, To 6 mL of a 1 M solution of SEH in ethyl acetate, 10 mL of isopropanol and 10 mL of water is added 1 mL (10 mmol) of $ 30 formalin. The mixture is stirred for 2 hours at room temperature to give a clear solution with a small amount of oily precipitate. After adding 1 g of sodium bisulfite to the solution, it is stirred for a further 2 hours, during which time the oily precipitate dissolves in the solution. The reaction mixture is poured into 300 ml of ethanol with vigorous stirring to give about 1.5 g of sodium 7- (Λ- (ο-Ν, N-bis (sodium sulfomethyl) aminomethylphenyl) -propionamido) -3- (tetrazolo (U , 5-b) Pyridazin-6-ylthiomethyl-3-cephem-U-carboxylate, which is collected by filtration, washed with three 50 ml portions of ethanol and dried in vacuo.

Claims (2)

An intermediate of the antibacterial 7- (o-aminomethylphenylacetamido and propionamido) -3- (tetrazolo / k) of the formula ch2nh2 ^ -ch2-s - (^ ^) = J1 I COOH wherein n is 1 or 2. 7-Acin-3- (tetrazoloyl-5-bipyridazin-6-ylthiomethyl) -3-cephemazin-6-ylthiomethyl) -3-cephem-U-carboxylic acid a carboxylic acid of the formula H „N — j - SS \ / N ^ T 2 N —N J-II COOH