GB2194790A

GB2194790A - Intermediates useful for preparing antibacterial agents

Info

Publication number: GB2194790A
Application number: GB08721347A
Authority: GB
Inventors: Haruhiro Yamashita; Masahisa Oka; Takayuki Naito; Jun Okumura
Original assignee: Bristol Myers Co
Current assignee: Bristol Myers Co
Priority date: 1984-04-09
Filing date: 1987-09-10
Publication date: 1988-03-16
Also published as: GB2194789A; AU4086285A; YU118987A; ES8706694A1; FR2563832A1; SE8901226D0; JPH0262557B2; IE58403B1; ES557060A0; HUT37622A; GB2157293B; NL8501002A; DE3512225C2; IE850866L; CA1276929C; AT388735B; LU85840A1; GB2157293A; SU1487814A3; JPH0350754B2

Description

1 1 1 GB2194790A 1

SPECIFICATION

Intermediates useful for preparing antibacterial agents Summary of the Invention This application relates to 7[2-(5-amino-1,2,4thiadiazol-3-yi)-2-(substituted)iminoacetamidol-3-[3(quaternaryammonio)-1propen-l-yil-3-cephem-4-carboxylates of the formula

N--T- C - CONH - - ', S 11 3:L 6N H H H 0 \ OR2 0 C000 CKH%-N =-Q 1 in which R' and R2 are as defined herein and -ON-=Q is a quaternary ammonio group as defined herein, and to salts and esters thereof. This invention also relates to processes for the preparation of the compounds of Formula 1, to pharmaceutical compositions containing at least one compound of Formula 1, and to intermediates in their preparation.

Background and Prior Art (A) U.S. Patent 4,390,534, issued June 28, 1983 to Tsutomu Teraji et aL, discloses cephem and cepham compounds of the formula

R1 N-S, _ - S 3 CONH R4 R5 \ 5 A N W wherein RI is amino or protected amino; R2 is hydrogen, acyl, optionally substituted aryl, substituted alkyl, alkenyl, alkynyl, optionally substituted cycloalkenyl or an 0- or S-containing 5membered heterocyclic ring substituted with oxo group(s); R3 is hydrogen or alkyl; R4 is hydro- gen, acyloxyalkyl, acylthioalkyl, optionally substituted pyridinioalkyl, optionally substituted heterocyclylthioalkyl, alkyl, halogen, hydroxy or optionally substituted thiazolioalkyl; and R5 is carboxy or protected carboxy; provided that R6 is COO- when R4 is optionally substituted pyridinioalkyl or optionally substituted thiazolioalkyl; and the dashed line indicates either a single or double bond.

European Patent Application No. 13,762, published August 6, 1980 is concordant thereto and has a similar disclosure.

U.S. Patents 4,381,299 (issued April 26, 2983), 4,331,665 (issued May 25, 1982) and 4,332,798 (issued June 1, 1982) each issued on parent applications of U.S. 4,390,534, and have similar disclosures. (13) European Patent Application No. 62,321, published October 13, 1982, discloses cephem compounds of the fprmula R1 N -CONH "S N ---1.1 S,,-' N H 0 \ 1R2 - 0 coo 9 wherein R1 is amino or protected amino; R2 is an optionally substituted lower aliphatic hydrocar55 bon group, or cycloalkenyl; and the group of the formula -NO 1 X - - 1 is an optionally substituted heterocyclic cation group containing more than one nitrogen atom; and pharmaceutically acceptable salts thereof. Also disclosed are intermediates of the formula The present Application has been divided out from Application No.85.08846 (21257293).

2 GB2194790A 2 N R1 -CONH -F, ' XE) 1 /- - ' S N H 1 -N (1) '1 OR2 0 R4 CH2 wherein R, and R2 are as defined above, R4 is a protected carboxyl group and X- is an acid 10 residue. (C) European Patent Application No. 74,653, published March 23, 1983, discloses cephem compounds of the formula Rl--H4-'-7--C-CONH 3 H S / N rCH 2N r O-R2 0 n 0aR c 0 wherein RI is amino or protected amino; R2 is an optionally substituted lower aliphatic hydrocarbon group, cyclo(lower)alkyl or cyclo (lower)alkenyl; R3 is (lower)alkylamino, N-protected(lower)alkylamino, di(lower)alkylamino, sulfo(lower)alkylam ino, hydroxy(lower)alkylamino, N-protected hydroxy(lower)alkylamino, acyloxy(lower)alkyl, (lower)alkoxy(lower)alkoxy(lower)aikyl, di(lower)alkylamino(lower)alkyl, (lower)alkylthio(lower)alkyl, (lower) alkylthio, (lower)alkoxy, (lower)alkoxy(lower)alkoxy, hydroxy(lower)alkoxy, acyl(lower)alkyl, hy droxy(lower)alkylthio, di(lower)alkylamino(lower)alkylthio, N-containing unsaturated 5-membered heterocyclic group, N-containing unsaturated 5-membered heterocyclicthio, or N-containing un saturated 5 or 6-membered heterocyclicthio(lower)alkyl Which may be substituted with suitable 30 substituent(s); and R4 is hydrogen or (lower)alkyl; or a salt thereof.

(D) U.S. Patent 4,332,800, issued June 1, 1982 to Tsutomu Teraji et aL, discloses intera alia compounds of the formula R' N--5-C -CONH - 5 H 11 -S/ -- H 6R2 t1 -% N CH l 2m.,X C00e wherein R' is amino or protected amino; R2 is (lower)alkyl and X is hydrogen or carbamoyl. (E) European Patent Application No. 47,977, published March 24, 1982, discloses cephem 45 compounds of the formula (0)m Am -T- C- CONH S 0 N N 1 F C-R2 CH2R1 coo (D wherein m is 0 or 1; Am is optionally substituted amino; T is a thiadiazolyl moiety (attached to the other groups by two of its carbon atoms); R2 is hydrogen, optionally substituted alky], cycloalkyl or optionally substituted carbamoyl; and R, is optionally substituted thiazolio, optionally substituted pyrazolio, tri(lower)aikylammonio or a pyridinio group of the formula -1 3 GB2194790A 3 Ra G) -H Rc'tRb in which Ra is substituted (lower)alkyl [the substituent being cycloalkyl, phenyl, hydroxy, alkoxy, 10 halogen, cyano, carbamoyl, carboxyl or suifo], (lower)alkenyl or carboxy- substituted (lower)aikenyi, (lower)alkylthio or carboxy-substituted (lower)aikyithio, amino or monosubstituted amino [the substituent being (lower)aiky]. (lower)alkanoyl or aminobenzenesuifonyll, diflower)alkylamino, substituted carbamoyl [the substituent being (lower)alkyl, hydroxy(lower)aiky], (lower)aikoxy, hydroxy or cyanol, di(lower)aikylcarbamoyi, thiocarbamoyl, cycloalky], phenyl, hydroxy, (lower)aikoxy, halo- 15 gen, (lower)aikoxycarbony], (lower)aikanoyloxy, (lower)aikanoyi, carboxyl, suifo, cyano, nitro or hydroxysulf,o(lower)aikyi; Rb is hydrogen or carbamoyl, or has the same meaning as Ra; and Re is hydrogen or has the same meaning as Ra; and salts thereof.

Although not formally related, European Patent Application No. 25,017, published March 11, 20 1981, has a similar disclosure. (F) European Patent Application No. 30,630, published June 24, 1981, discloses 3-vinyl cephem compounds of the formula

R1-A-CONH PN S - C"CH2 0 R2 wherein RI is an optionally protected amino-substituted heterocyclic group which may also have halogen, or a group of the formula R3 SO 2 H N ",,C, y in which R:3 is (lower)alkyl; R2 is carboxy or protected carboxy; and A is lower alkylene which may have a substituent selected from amino, protected amino, hydroxy, oxo and a group of the formula =N-OR4, wherein R4 is hydrogen, cyclo(lower)alkenyl, (lower)alkynyl, (lower)alkenyl [optionally substituted by carboxy or protected carboxyl, (lower)alkyl [optionally substituted by one or more of carboxy,- protected carboxy, amino, protected amino, cyano, phosphono, protected 45 phosphono and a heterocyclic group which itself may be substituted]; and salts thereof. This application specifically discloses compounds of the formula N C-CONH S -7 11 H2 N---,\ S /N N \0)ii CH CH2 R4 0 in which OR4 is methoxy, carboxymethoxy, tert-butoxycarbonyimethoxy or 1- tert-butoxycarbonyl- 55 ethoxy. (G) U.K. Patent Specification No. 1,399, 086 published June 25, 1975, contains a generic disclosure encompassing a vast number of cephalosporins of the formula

4 GB2194790A 4 R- C- 10-NH H\ H p oe 0 1 p COON wherein R is hydrogen or an organic group, Ra is an etherifying monovalent organic group linked 10 to the oxygen through a carbon atoms B is S or S'O' and P is an organic group. In one embodiment, P may be inter alia a vinyl group of the formula R3 -CH= C "\R4 in which R3 and R4 independently may be hydrogen, nitrile, (lower)alkoxycarbonyl, or substituted 25 or unsubstituted aliphatic, cycloaliphatic, araliphatic or aromatic. However, the 5-amino-1,2, 4thiadiazol-3-yl group is not identified as a possible R substituent and there is no disclosure or suggestion that P may be a quaternary ammoniosubstituted propenyl group. U.S. Patent 3,971,778 and its divisionals Nos, 4,024,133, 4,024,137, 4,064,346, 4,033,950, 4,079,178, 4,091,209, 4,092, 477 and 4,093,803 have similar disclosures.

(H) Eurpean Patent Application No. 88,385, published September 14, 1983, discloses corn- 30 pounds of the formula Rlc- CONH - 0 H N R3 W R4 in which RI is (unsubstituted) thiadiazolyl; R2 is carboxy(lower)alkyl or protected carboxy(lower)al- 40 kyl; R3 is hydrogen, halogen or (lower)alkenyl; and R4 is carboxy or protected carboxy. Although 1propenyl is listed as one of the possible meanings of R3, the application only exemplifies compounds where R3 is hydrogen, chloro or vinyl. (1) U.S. Patent No. 4,307,233 issued to Daniel Farge et al. on December 22, 1981, discloses 45 inter alia, 3-vinylcephalosporin derivatives of the formula N C- CONN S a N2N"S) N \ OR5 1/ R3 COON C"CH-H" R4 in which R5 inter alia may be alkyl, vinyl, cyanomethyl or a protective group such as 2 methoxyprop-2-yl, and R3 and R4 are alkyl groups (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R3 and R4, taken together with the nitrogen to which they are attached, may form a saturated heterocyclic ring of 5 or 6 members, optionally containing another hetero-atom selected from N, 0 and S, and optionally substituted by an alkyl group. The compounds are useful as intermediates in the preparation of 3-thiovinyl 60 cephalosporin derivatives. There is no disclosure or suggestion of a 5- amino-1,2,4-thiadiazol-3-yl moiety in place of the 2-aminothiazol-4-yi substituent or of a quaternary ammonio-substituted propenyl moiety for the 3-substituent. Published United Kingdom Patent Application No.

2,051,062 is concordant thereto and has a similar disclosure.

(J) European Patent Application No. 53,537, published June 9, 1982, discloses inter alia, 3- 65 -1 GB2194790A 5 vinyleephalosporin derivatives of the formula c N C _CONH S 11 R3 ITN CH-N H2N \ S 0 \-C-COORC COOR2 CH Z\ b R5 R5 in which Ra and R b are the same or different and are hydrogen or alkyl, or taken together, form 5 an alkylene group containing 2 or 3 carbon atoms, Rc is an acid protecting group, R2 is an acid protecting group such as an ester, R, and R4 are the same or different and are hydrogen, alkyl (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R3 and IR, taken together with the nitrogen to which they are attached, may form a saturated heterocyclic ring of 5 or 6 members, optionally containing another hetero-atom selected from N, 0 and S, and optionally substituted by an alkyl group. The compounds are useful as intermedi- ates in the preparation of 3-thiovinyl cephalosporin derivatives. There is no disclosure or sugges- 20 tion of a 5-amino-1,2,4-thiadiazol-3-yi moiety in place of the 2- aminothiazol-4-yl substituent or of a quaternary ammonio-substituted propenyl group for the 3-substituent.

U.S. 4,423,214 is concordant thereto and has a similar disclosure.

(K) European Patent Application No. 53,074, published June 2, 1982, generically discloses a vast number of 3-vinylcephalosporin derivatives of the formula wn Cla-NH R 1 3N ' ' - CH= C- R3 0 COOR c) 2a wherein R',. (in one of several embodiments) may be N 1-10- --TN I U N2N ' -.S" "OR5 in which R, inter alia may be hydrogen, alkyl, vinyl, cyanomethyl, an oxime-protecting group such as trityl, etc., or a group of the formula c R a/ -,\ R b 5 5 in which Ra and R b are the same or different, and may be hydrogen, alkyl or, taken together, an 5 alkylene radical of 2 or 3 carbon atoms, and RC5 is hydrogen or an acid- protecting radical; R02a is hydrogen or an acid-protecting radical such as methoxymethyl; R' (in one of several embodi ments) may be a methyl group substituted by a 5- or 6-membered aromatic heterocyclic ring containing a single hereto atom, such as 2- or 3-pyridy], 2- or 3-thienyl or 2- or 3-furyl, and R. 55 is a group of the formula R,SO,O- in which R4 may be alkyl, trihalomethyl or optionally substituted phenyl.

These compounds are stated to be intermediates in the preparation of compounds in which the 3-substituent is a group of the formula 6 GB2194790A 6 R 0 i -CH= C-SR which are stated to have antibacterial activity.

Although this patent includes the possibility of R being a methyl group substituted by an Ncontaining heterocyclic ring, in both the intermediates and final products (thus giving a heterocyclic-substituted propenyl moiety), it teaches only that the heterocyclic ring is attached via one of its carbon atoms. Thus, there is no suggestion of a quaternary ammoniosubstituted propenyl 10 group. The reference exemplifies R in the intermediates and final products only as methyl. Further, in both the intermediates and final product, the propenyl group must contain a second substituent (-0,SR4 or - SR, respectively). Also there is no disclosure or suggestion of a 5amino- 1,2,4-thiadiazol-3-yi moiety in place of the 2-aminothiazol-4-yl substituent.

(L) European Patent Application No. 53,538, published June 9, 1982, discloses, inter alla, 3- 15 vinylcephalosporin intermediates of the formula H C-CONN N\OR5 H2 S375 0)- -- CH=CH- R3 in which n is 0 or 1, R5 is hydrogen, alkyl, vinyl, cyanomethyl or an oxime-protecting group, and R3 is halogen. There is no disclosure or suggestion of a 5-amino-1,2,4-thiadiazol-3-yi moiety in place of the 2- aminothiazol-4-yl substituent, and no disclosure or suggestion of a 3- halo-lpropen-l-yl substituent in the 3-position.

Complete Disclosure

This application relates to novel cephalosporin derivatives which are potent antibacterial agents. More particularly, it relates to compounds of the formula N r 5 35 A 11 11 A N OHN S \ OR2 wherein R' is hydrogen or a conventional amino-protecting group, R2 is hydrogen, a straight or branched chain alkyl group containing from 1 to 4 carbon atoms, a cycloalkyl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula 45 R4 R4 COOH -c- CH=M-0 -C'-C=_ C-0 1 1 or RS 50 4 -C- COOH ' 1 RS in which R3 is hydrogen, (lower)aikyl or carboxyl, X is halogen, hydroxy or (lower)alkoxy, and R4 and R5 are each independently hydrogen, methyl or ethyl, or R4 and R5, taken together with the 60 carbon atom to which they are attached, may'be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and 7 GB2194790A 7 is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts of physiologically hydrolyzable esters thereof. Also included within the scope of the invention are the solvates (including hydrates) of the compounds of Formula 1, as well as the tautomeric forms of the compounds of Formula 1, e.g. the 2-iminothiazolin-4-yl form of the 2- aminothiazol-4-yi moiety.

In another aspect, this application relates to a process for the preparation of the compounds of Formula I and to certain intermediates in their preparation.

As shown in the structural formula, the compounds of Formula I have the "syn" or -Zconfiguration with respect to the alkoxyimino group. Because the compounds are geometric isomers, some of the "anti" isomer may also be present. This invention comprises compounds of Formula I containing at least 90% of the "syn" isomer. Preferably the compounds of Formula I are "syn" isomers which are essentially free of the corresponding "anti" isomers.

In addition to geometric isomers possible with respect to the alkoxyimino group, the compounds of Formula I (and the intermediates of Formulae Vill and IX) also form geometric (cis and trans) isomers about the double bond of the propenyl group. Both the cis ("Z") and trans ("E") isomers of these compounds are specifically included within the scope of this invention.

The nontoxic pharmaceutically acceptable salts of the compounds of Formula I include salts with mineral acids such as hydrochloric, hydrobromic, phosphoric and sulfuric, or with organic carboxylic acids or sulfonic acids such as acetic, trifluoroacetic, citric, formic, maleic, oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic, malic, methanesulfonic, benzenesulfonic, p-toluenesulfonic and other acids known and used in the penicillin and cephalosporin arts.

Preparation of these acid addition salts is carried out by conventional techniques.

Examples of physiologically hydrolyzable esters of the compounds of Formula I include indanyl, phthalidyl, methoxymethyl, acetoxymethyl, pivaloyloxymethyl, glycycloxymethyl, phenylglycyloxy methyl, 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyI and other physiologically hydrolyzable esters known and used in the penicillin and cephalosporin arts. Such esters are prepared by conven tional techniques known in the art.

The compounds of Formula I in which R1 is hydrogen exhibit high antibacterial activity against various Gram-positive and Gram-negative bacteria, and are useful in the treatment of bacterial infections in animals, including man. The compounds of Formula I may be formulated for parenteral use in a conventional manner utilizing known pharmaceutical carriers ane excipients, and may be presented in unit dosage form or in multi-dosage containers. The compositions may be in the form of solutions, suspensions or emulsions in oily or aqueous vehicles, and may contain conventional dispersing, suspending or stabilizing agents. The compositions may also be in the form of a dry powder for reconstitution before use, e.g. with sterile, pyrogen-free water.

The compounds of Formula I may also be formulated as suppositories utilizing conventional suppository bases such as cocoa butter or other glycerides. The compounds of this invention may, if desired, be administered in combination with other antibiotics such as penicillins or other cephalosporins.

When provided in unit dosage forms the compositions will preferably contain from about 50 to about 1500 mg of the active ingredient of Formula 1. The dosage of the compounds of Formula I is dependent on such factors as the weight and age of the patient as well as the 45 particular nature and severity of the disease, and is within the discretion of the physician.

However, the dosage for adult human treatment will usually be in the range of from about 500 to about 5000 mg per day, depending on the frequency and route of administration. When administered intramuscularly or intravenously to an adult human, a total dosage of from about 750 to about 3000 mg per day, in divided doses, normally will be sufficient, although higher daily doses of some of the compounds may be desirable in the case of Pseudornonas infections.

The quaternary ammonio group of the formula % = 0 may be acyclic, cyclic, or a combination of the two, and may contain one or more additional hetero atoms selected from nitrogen, sulfur and oxygen.

An example of an acyclic quaternary ammonio group is a group of the formula 8 GB2194790A 8 OR6 1 -N - R7 1 R8 in which R6, R 7 and R8 may be the same or different and may, for example, be (lower)alkyl or substituted (lower)alkyl in which the substituents are, for example, halogen, amino with the provision that the amino group may not be on an a-carbon, hydroxy with the provision that the 10 hydroxy group may not be on an a-carbon, (lower)alkoxy with the provision that the alkoxy group may not be on an a-carbon, (lower)alkylthio, (lower)alkylamino, di(lower)alkylamino, carbamoyl, (lower)alkenyl, phenyl(lower)alkyl, phenyl or substituted phenyl (in which the substituents may be, for example, halogen, hydroxy, amino, (lower)alkylamino, di(lower)aikylamino, acylamino, (lower)alkyl, (lower)alkylthio, (lower)alkoxy or the like).

Examples of cyclic quaternary ammonio groups are fully unsaturated monocyclic heterocyclic ring systems, and bicyclic heterocyclic ring systems in which at least one N-containing ring is fully unsaturated. Suitable cyclic quaternary ammonio ring systems include, for example, those of the formulae 20 0 R9 TJ--3, R9 G R9 0 00 25 N R9 30 R9 R9 j 10 U','S R-10 0 N-N R9 (9 N R9 35 R9 FR10 -+!-RlO u,'.1 0 --, S,-' H,S,-,N 0 AI(O R9 40 R10 R9 R10 N - WC LO w)- 0 N 45 1 G) N R9 0 N::, N c j 50 N S R9 R10 1 E) R10 (D - H N n,., 6o R19 and the like, in which R9 and R") are the same or different and may be, for example, hydrogen, halogen, amino, (lower)aiky], (lower)alkenyl, (lower)alkyithio, carboxy, hydroxy, (lower)aikoxy, (lower)aikoxy(lower)aikyl, halo(lower)alkyi, hydroxy(lower)aikyi, amino(lower)alkyi, (lower)aikylami- no(lower)alkyi, di(lower)aikylamino(lower)aikyi, (lower)aikylamino, di(lower)aikylamino, carboxy(low- 65 9 GB2194790A 9 er)aikyl, carboxy(lower)aikylamino, carboxy(lower)aikyithio, carbamoyl, N- (iower)aikylcarbamoyl, formylamino, acylamino, acyloxy, phenyl, pyridyl, amidino, quanidino and the like. Where the structure of the heterocyclic ring permits, R9 and R10, taken together, may be an alkylene group containing from 3 to 5 carbon atoms, e.g. propylene.

Examples of combined acyclic/cyclic quaternary ammonio groups include, for example, those of the formulae R12 02 0 //--R12 (D R12 R11 R11 0 WY/H\,,5 10 R12 of-)l R12 0 /-v R12 15 N 0 m S -N NH RT/ RI1 20 0 (D -N R12.7 N R12 R11 R11 25 W-\ 0 -N N N N-(lower)alkyl -N R12 R11 R1 2 30 R11 Ril and the like, in which R" may be, for example, (lower)alkyl, (lower)aikoxy(lower)aikyi, hydroxy (lower)aikyl with the provision that the hydroxy may not be on an a- carbon, carboxy(lower)aikyi, 35 amino(lower)aikyl with the provision that the amino may not be on an acarbon, (lower)aikenyi, halo(lower)alky], allyl and the like, and R12 may be, for example, hydrogen, hydroxy, halogen, (lower)aikyi, hydroxy (lower) a] kyl, (lower)aikoxy(lower)alkyi, halo(lower)aikyi, amino(lower)alkyi, (lower)aikoxy, (lower) alkylthio, (lower)aikenyi, amino, (lower) alkyla mino, di(lower)alkylamino, acy 40]amino, acyloxy, carbamoyl, amidino(lower)aikyi, phenyl, pyridyl, amidino, quanidino and the like.

Preferred quaternary-ammonio groups are those of the formulae RB G R16 G) 1 N -N-R14 R16 R R17 R17 50 (D 1 0 J R18 - N -(CH2)n and -N N 18 SAR21 R R20 55 wherein R 13, R 14 and R15 are the same of different and are (lower)alkyi, (lower)aikeny], amino(low er)aikyl with the provision that the amino may not be on an a-carbon, or hydroxy(lower)aikyl with the provision that the hydroxy group may not be on an a-carbon; R 16 is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkylthio, amino, (lower)aikylamino, diflower- 60 alkylamino, formylamino, (lower)alkanoylamino, carboxy, hydroxy, carboxy(lower)aikyi, carboxy(( lower)alkylthio, hydroxy(lower)aikyl, halo(lower)ai.kyi, amino(lower)alkyl, (lower)alkoxy(lower)alkyi, carbamoyl or N-(iower)aikylearbamoyi, or R 16 may represent a divalent alkylene group having 3 to carbon atoms; R17 is (lower)alkyi, (lower) alkoxy(lower)aiky], halo(lower)aikyi, allyl hydroxy(lower)aikyl with the 65 GB2194790A 10 provision that the hydroxy group is not on the a-carbon, amino(lower)aikyl with the provision that the amino group is not on the a-carbon, or phenyl (lower)alkyl; R18 is hydrogen, (lower)aikyl, (lower)aikoxy., (lower)alkoxy(lower)aiky], (lower)aikyithio, amino, (lower)aikylamino, di(lower)aikylamino, carboxy, hydroxy, carboxy(lower)aikyi, hydroxy(lower)aikyi, amino(lower)aikyi, formylamino, (lower)alkanoylamino, carbamoyl or N- (iower)aikylcarbamoy]; 5 n is an integer of from 1 to 3, inclusive; Z is CH2 or, when n, is 2, Z also may be S, 0 or N-R19, in which R19 is hydrogen or (lower)aiky]; and R20 and R21 are the same or different and are hydrogen, (lower)alky], (lower)alkoxy, (lower)alkyl thio, amino, (lower)alkylamino, di(lower)alkylamino, carboxy, hydroxy, hydroxy(lower)alkyi, amino- 10 (lower)alkyi, (lower)alkoxy(lower)alkyi, carboxy(lower)aiky], carboxy(lower)alkylamino, (lower)alka noylamino, carboxy(lower)aikanoylamino, carbamoyl or W(lower)alkylcarbarnoyl.

Particulary preferred quaternary ammonio groups are W(lower)alkylpyrrolidinio (and especially N methylpyrrolidinio), tri(lower)alkylammonio (and especially trimethylammonio), pyridinio, aminopyri dinio, formyiaminopyridinio, carbamoylpyridinio, amino(lower)alkylpyridinio, carboxypyridinio, hydroxy(lower)aikylpyridinio, N-(lower)aikylcarbamoylpyridinio, (lower)alkylenepyridinio, 2-methylthiazolio and 2-amino-5-thiazolo[4,5clpyridinio.

In the compounds of Formula 1, particularly preferred values of R 2 are (lower)alkyl (and especially methyl) cycloalkyl containing from 3 to 5 carbon atoms, 1-carboxycycloalk-l-yl containing from 3 to 5 carbon atoms, allyl, propargyl and carboxy(lower)aikyl (and especially 2-carboxyprop2-yl). The most preferred compounds of the invention are (1) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3(trimethy lammonio)-1- propen- 1 -yll-3-cephem4-carboxylate, (2) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(1methyl pyrrolidinio)-1- propen-l-yi]-3-cephem-4-carboxylate, (3) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3pyridinio -l-propen-l-yil- 3-cephem-4-carboxylate, (4) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamido]-3-[3-(3aminop yridinio)- 1 -pro pen- 1 -y11-3-cephem-4-carboxylate, (5) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3-(3formyl aminopyridinio)- 30 1 -propen- 1 -y11-3-cephem-4-carboxylate, (6) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3-(3aminom ethylpyridi- nio)- 1 -propen- 1 -yi]-3-cephem-4-carboxylate, (7) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3carbam oyipyridinio)-1- propen- 1 -y11-3-cephem-4-carboxylate, (8) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4carbam oylpyridinio)-1- propen- 1 -y11-3-cephem-4-carboxylate, (9) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3-(2methyi thiazolio)-1- propen- 1 -y11-3-cephem-4-carboxylate, (10) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3(2-amino-5 -thia- 40 zolo[4,5-c]pyridinio)- 1 -propen- 1 -yi]-3-cephem-4-carboxylate, (11) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3(4-hydrox ymethylpyridi- nio)- 1 -propen- 1 -y11-3-cephem-4-carboxylate, (12) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3(3-hydrox ymethylpyridi- nio)- 1 -propen- 1 -y11-3-cephem-4-carboxylate, (13) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3(4-IN-met hyicarbam- oyllpyridinio)- 1 -propen- 1 -yi]-3-cephem-4-carboxylate, (14) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamido]-3-[3(2,3-prop yienepyridi- nio)-1-propen-l-yil-3-cephem-4-carboxylate,.

(15) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-ethoxyiminoacetamido]-3-[3-(4carbamo yipyridinio)-1- 50 propen- 1 -y11-3-cephe ' m-4-carboxylate, (16) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-cyclopentyloxyiminoacetamidol3-[3-(4 -carbamoylpy- ridinio)- 1 -propen- 1 -y11-3-cephem-4-carboxylate, (17) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-allyloxyiminoacetamidol-3-[3(4-carba moylpyridinio)- 1 -propen- 1 -yi]-3-cephem-4-carboxylate, (18) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetamidol-3[3-(4-c arbamoylpyridi- nio)-1-propen-l-yil-3-cephem-4-carboxylate, (19) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3(4-carbox ypyridinio)-1- propen- 1 -yi]-3-cephem-4-carboxylate, (10) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-ethoxyiminoacetamidol-3-[3-(4carboxy pyridinio)-2- 60 propen- 1 -y11-3-cephem-4-carboxylate, (21) 7-[2-(5-amino-1,2,4thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3-(3carbox ymethylpyridi- nio)-1-propen-l-yil-3-cephem-4-carboxylate and (22) 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3(4-carbox ymethyithio- pyridinio)-1-propen-l-yi]-3-cephem-4-carboxylate.

11 GB2194790A 11 The numbering system utilized herein for the various reactants, intermediates and final products is as follows:

- F 1ROM&n I(Arabic Numeral i ú appropriate) e Letter 5 (if appropriate) The Roman Numeral designates whether the compound is a final product [11 or an intermediate or 10 other reactant [all other Roman Numerals]. The Arabic Numerals and Letters are not used in those instances where the overall class (genus) of compounds is meant.

The Arabic Numeral designates the particular meaning of substituent R2. If the particular R2 group contains a carboxyl group which is protected by a conventional carboxyl-protecting group, a---prime-(') is used after the Arabic Numeral to indicate this fact. No---prime-is used if the carboxyl group is unprotected. A---prime-also is used with the generic R2 substituent (i.e. RT) when generically referring to an R2 group containing a protected carboxyl group.

The Letter at the end of the compound number refers to the particular meaning of the quaternary ammonio group % = 0 For convenience, the Arabic Numerals and Letters assigned to some of the preferred R2 25 groups and quaternary ammonio groups are set forth below. Arabic Numeral 1 2 3 R 2 - methyl ethyl allyl 4 propargyl cyclopentyl m 12

1 L K H 0 p Q GB2194790A 12 - 6) Letter -- _MO A 1-rethylpyrrolidinio a pyridinio 5 c 2-amino-5-thiazolo[4#5-CIPYridiniO D trimethylamonio z 3-aminopyridinio 10 3-foroylazinopyridinio G 3-carbasnoylpyridinio 4-carbarnoylpyridinio 15 3-amincmethylpyridinio j 2-methylthiazolio 3-hydroxymethylpyridinio 4-hydroxymethylpyridinio 4-(Nmethylcarhamoyl)pyridinio 4-carboxypyridinio 2#3-propylenepyridinio 3carboxymethylpyridinio 4-ca rboxymethyl thiopyri din io In the primary evaluation of the compounds of this invention, the Minimum Inhibitory Concen trations (MIC's) of the compounds were determined by the two-fold serial agar dilution method 35 in Mueller-Hinton agar against 32 strains of test organisms in six groups. The geometric means of the MIC's determined in these tests are shown in Table 1.

13 GB2194790A 13 Table 1

1 15 Conf ig. Geometric mean of IMIC (meglai) at 1 CIRP6. double (G+).la (G+)-Ib M-)-U M-)-1b M-)-11 (G-)-III bond (5) (5) (5) (5) (5) (7) 1-1A EIZW1/1 0.26 0.70 0.05 0.15 0.23 2.4 I-1A EIZa7/1 0.13 0.35 0.029 0.05 0.17 1.4 I-1B - E 0.20 0.40 0.016 0.044 0.11 1.6 1-18 ' E/Z-114 0.35 0.80 0.05 0.11 0.35 3.5 I-lc z 0.10 0.20 0.0071 0.033 0.087 3.8 1-1D ú/z-1/1 0.61 1.4 0.10 0.26 0.46 2.4 1-1D E/Z-1011 0.30 0.53 0.05 0.076 0.26 1.3 1-1E 0.20 0.40 0.0094 0.029 0.10 1.4 1-1F 0.15 0.40 0.0094 0.033 0.099 1.2 1-1G 0.20 0.35 0.0094 0.033 O.iO 1.4 1-15 0.20 0.40 0.013 0.043 0.10 0.97 E 0.80 1.6 0.10 0.20 0.69 3.1 E 0.17 0.35 0.025 0.076 0.15 1.6 I-1x z 0.35 0.80 0.029 0.044 0.20 3.5 I-1L 0.26 0.61 0.029 0.088 0.15 2.6 I-1m 0.35 0.70 0.029 0.10 0.17 2.3 1-1N EIZ-7/1 1.2 1.6 0.013 0.066 0.30 5.7 I-10 z 0.17 0.35 0.029 0.033 0.11 14 1-2E z 0.20 0.40 0.014 0.057 0.15 1.4 I-2N z 1.2 2.1 0.016 0.11 0.35 4.7 I-2N Z 1.4 3.1 0.044 0.15 0.69 10 1-3E z 0.23 0.40 0.057 0.10 0.52 1.9 1-4m 0.26 0.46 0.066 0.11 0.60 2.6 I-5m 0.13 0.40 0.20 0.46 2.1 4.2 I-1p 0.8 1.6 0.013 0.087 0.34 14 0.7 0.92 0.0095 0.044 0.23 14 14 GB2194790A 14 (G+)-1a Penicillin-sensitive S. aureus (5 strains) (G+)-Ib Penicillin-resistant S. aureus (5 strains) (G -)-fa Cephalothin-se-nsitive E. coli (2 strains), KI.

pneumoniae (1 strain) and Pr. mirabilis (2 strains) 5 (G-)-Ib Cephalothin-resistant E. coli (3 strains) and KI.

pneumoniae (2 strains) (G-)-Ii M. morganfi (1 strain), Ent. cloacae (2 strains) and Ser. marcescens (2 strains) (G-)-111 Ps. aeruginosa (7 strains) 10 Table 2, below, gives the Protective Dose,, (PD51) in mice for a number of the compounds of Formula 1 against selected microorganisms. Table 3 gives blood levels of various compounds of Formula 1 upon intramuscular administration of the test compounds to mice at a dosage of 20 mg/kg.

Table 2

PD so (Inglkg) 20 5. aurcus E. Coli P. aeruginosa CE2d. No. smith Juhl A9843A 25 1-1B 0.44 0.028 7.7 1-1B 0.65 0.072 NT I-lc 0.22 0.013 NT I-IG 0.96 0.021 5.92 30 1-15 0.39 0.015 3.9 0.35 0.029 NT I-1x 0.53 NT NT 35 1-1m 0.96 NT INT 1-1N 2.0 NT NT 1-10 0.26 0.17 HT 1-2N 5.0 HT NT 40 NT a Not Tested GB2194790A 15 Table 3

Cleax T112 AUC 5 Cmd. No. (MC g /A1 (min) (incg br/wil) I-1B 17 21 11 I-lc 21 32 18 1-1D 20 19 11 10 I-1H 23 16 14 1-11 19 16 9.7 I-1x 24 14 14 15 I-im 20 23 14 1-1N 24 19 18 1-10 28 32 17 1-2N 22 20 12 20 1-21E 19 47 25 1-4E 27 22 16 -SE 22 32 is 25 In another aspect, this invention relates to processes for the preparation of the compounds of Formula 1. The preferred procedures are shown below in Reaction Schemes la, 1b and lc, while an alternative procedure is shown in Reaction Scheme 2. The abbreviation --Ph- represents the 30 phenyl group. Thus, the -CH(Ph)2 moiety is the benzhydryl group, which is a preferred carboxy] protecting group. When R2 contains a carboxyl group, it is desirable to protect the carboxyl group with a conventional carboxyl-protecting group such as the t-butyl moiety. Y represents chloro, bromo or iodo.

16 GB2194790A 16 Reaction Scheme la 5 PN:;ICH2Ct 11 COOCH(PM2 H-T-F-COON 10 - 1 N2N S,,,' N L, OR2 Ill 15 ,- 5 -, N E-CONN \ 11 - 1 H N\ 2H//\ S\ OR2 COOCH(M)7 Nal or KI 25 H 1 - CO N H 1,-5 -'/\ S/ N N \ OR2 0)- N CH21 v 1 COOIN(PM2 P(PN3 P1ph)3 N C-CONH S Y(D I -F, CH2 P(Ph)3 vi HXO\ S/ OR2 0 N OOCH(Ph)2 base 45 C - CONH S N \\ 11 H2- S/ N H\ OR2 0 H CH 2P(PW3 V11 COOCH(PN2 C1CH2CHO 55 - N C-CONH "'s if N N N CH=CHCHZCI Vill 11 H28 5 "" \ OR2 0 -F" COO C H (Ph)2 17 GB2194790A 17 IX Nal or KI N H S/ 'OR2 CK H2N"' COOCH(PM2 NCH21 ,- R HN 'R' (secondary amine) N--\.-t -CONN 5 HAI\ S/ N N " OR2 0 WINCH2-N \R d?OOCH (M2 X R 11 v N - CONH - S Y e I -11 / N H-' 2 N CH=CHCH G =O; 1 "" H2N S OR 0 TN, 2-m- 0 =-N xl (terti ary ami ne) X11 COOCH (PM2 deblock N C-CONH 5 45 \\1 N H N (D Ir-11\ 11/ H2H \CR2 0 CH= IN %- NE 0 3PN - 100 G) 50 18 GB2194790A 18 Reaction Scheme 'I a shows two alternate means of going from Compound IX to Compound XIL The direct route, utilizing a tertiary amine (M), is applicable for the preparation of all compounds of Formula 1. The indirect rou ' te, via Compound X, utilizes a secondary amine as reactant, and is quaternized in the following step. The secondary amine WNH may be acyclic (e.g. dimethylamine) or cyclic (e.g. pyrrolidine), and this indirect procedure therefore is suitable for the preparation of compounds of Formula 1 in which the quaternary ammonio group is acyclic or -mixed- acyclic/cyclic. This indirect route is not suitable for the preparation of compounds of Formula 1 wherein the quaternary nitrogen is in a fully unsaturated heterocyclic ring (e.g. pyridinio, thiazolio, 2-amino-5thiazolo[4,5-c]pyridinio, and the like).

19 GB2194790A 19 Reaction Scheme lb II rkNO 5 t==:yf CH =N X111 10 G 7Mfl,' 0 C02C1 COO C H (M2 Nal KI G CH = NI T:N 5 CH 21 XIV 0 O0CH(PM2 P(PN3 P(PN3 1 - 30 (3-CH = N - (D y (E) 35 H CH2P(Phh xv base COOCKN2 40 H 3 N 0 N WP(PM3 COO CH(M2 M GB2194790A 20 C1CH2CHO 5 S 0 N 9-- CH="CHCH2CI XVII 10 COOCH (Ph)2 Girard Reagent T or H2N S HC1 15 0 CHCHCH2CI XVIII OOCH (Ph)2 20 N -,T.- -CUOH III H2N-'4\/S/N N\ OR2 25 V,,, as in scheme W'" 1 Reaction Scheme lb is a variation of Reaction Scheme 1 a in that the 7- amino group of the starting material (11) is protected as a Schiff base during most of the reaction steps, and the 30 desired 7-side chain acid is added later in the synthesis. Otherwise, the general procedure is similar.

G62194790A 21 (yeg=h 5 Reaction Schen"O 1 c cg=eg-ú"2ú1)(VII % al or M 1, P c" \--==i WA McondarY amme) r- c"=% /R XX XIX z XXI 0 XXII as# Xl Rertiary amine) x-acitation with K\ 92 0 cw G-D 1 G) is 22 GB2194790A 22 Reaction Scheme 'I c is a further variation of Reaction Scheme lb. In Reaction Schemes 'I a and 1 b, quaternization of the 3-side chain is the last step, but in Reaction Scheme 1 c the last step is acylation of the 7amino group. The relationship between Reaction Schemes l a, l b and 'I c is shown in the following flowchart.

N 5 5 1 N \b 1 1 0PN CM2C1 c 10 11 COOMPM2 N -CONH- 17-7 S TN,V - _CH' H2N A N ''OR2 0 CH2C' \-=ry" 91r_ CH2C' 15 IV COOCH(Ph12 X111 COOCH(PhI2 lb 1C 20 N---r C- CONN S 7-N-Acylation I J: - N lb N S"N N\OR2 CHrCHCH2C1 0 CHCH2C1 Vill COOCH(Ph)2 XVIII COOCH(Ph)2 la Quaternization lc Guarternization 30 1b Deblocking 7-H-Acylation 1 Deblocking Compound 1 lc N -1" 5 N 0;:-,: CH=CHCHiN-=0 35 XXII COC In Reaction Schemes l a, lb and 1c, the benzhydryl group was shown as the preferred carboxyl-protecting group. It will be appreciated by those skilled in the art that other carboxyl- 40 protecting groups, well-known in the art, may be used. The acylating acid III may be used in the form of a derivative such as its acid halide, activated ester, mixed acid anhydride, etc., all of which are well-known in the art. We prefer to utilize it in the form of its acid chloride. Acylating acid III also may have its amino group protected by any of the common amino-protecting groups, e.g. N-trityl, N-formyl, or the like. The base used to convert the phosphoniurn iodide (VI 45 or XV) to give the phosphorylide (VII or XVI) may be NaOH, Na,CO,, IRA- 410 (OH-) resin, IRA(CO,) resin, or the like, or a mixture thereof. The chloroacetaldehyde used to convert the phosphorylide Vil to the 3-chloropropenyl-3-cephem compound Vill (or Compound XVI to Com pound XVII) may be the commercially available 40-50% aqueous solution, a distilled solution (e.g. 70%) or the anhydrous aldehyde.

We have found that Compound Vill prepared from Compound VII (Scheme la) typically had a Z:E ratio of about 2:1 at the propenyl double bond. Compound Vill prepared from Compound XVIII (Scheme lb), on the other hand, typically was almost exclusively the Z isomer. The difference may not be in the route used, but in the conditions utilized in the Wittig reaction (VII to Vill or XVI to XVII). We have also found that the use of an appropriate silyl reagent such as 55 N,O-bis(trimethylsilyl)acetamide in the Wittig reaction (Vil to Vill in Scheme la and XVI to XVII in Scheme lb) caused improvement of the yields and purity of Vill and XVIL The reaction is preferably carried out with 2-5 equivalents of the silyl reagent. When the chloropropenyl ce phem (VIII) was reacted with Nal in acetone to give the iodopropenyl cephem (IX), the double bond in the propenyl group was isomerized from Z to E during the iodination. A short reaction 60 period retained the configuration of the parent Compound Vill to a large extent, while a long reaction period gave primarily the E isomer of Compound IX. However, an excessive reaction time at high temperature gives a lower purity compound IX. We find that about 10 minutes at 25'C and 2 hours at 5'C gives pure IX in good yield. When utilizing Reaction Scheme Ic, we have found that, when iodinatIng compound XIV with Nal, a purer compound is obtained if the 65 23 GB2194790A acetone solution is diluted with CC14 when iodination is essentially completed, and the isomeriza tion portion of the reaction is conducted in the acetone-CCI, mixture. When iodination of the chloropropenyl cephem (XVII) to the iodopropenyl cephern (M) was performed with KI in DMF, the isomerization of the double bond from Z to E proceeded as fast as the iodination did. The whole reaction completed within 45 minutes at room temperature to give pure XIX without 5 dilution with CCI, in the course of the reaction.

Compound X11 normally was deblocked without purification, and the final product (1) was purified by reverse phase column chromatography utilizing a glass column containing the packing removed from a Waters' Associates PrepPAK-500/C1. cartridge.

24 GB2194790A 24 Reaction Scheme 2 ll c - COHN --P:::N 5 0 -HN J, 5,0 H N \OR2 0 N C"CH-CH21 Mill CO0CH(M7 H S xxlv Rl - HNJ SA H., n 1/1 OR2 0 COOCH(Phh 0 1 0 N ---F C- CONH - 1 11 1, N 0 N ' E0 xxv op N 0 CH=CH-CH2 RIAN A, S.,N N,' 0 R2 CDOCH(PN.) 30 Reduction 35 R1AN k S,.A N 'OR2 N---Ti- C - CC N H ','S (D XXVI CO0CH(PM2 deblock N C- CONH 'A J1111 1 N2N S ', N N \\ OR2 0 - N_. (D 1 oge CH=CH2"a GB2194790A 25 Reaction Scheme 2, shown in brief outline form above, is similar to Reaction Scheme 1 a except that Compound XXIII (equivalent to Compound IX of Reaction Scheme 'I a) is converted to its S-oxide prior to quaternization. Compound XXV is subsequently reduced, and the remainder of Reaction Scheme 2 is as Reaction Scheme 1 a. In Reaction Scheme 2, it is preferred to protect the amino group of the 7-side chain with a known amino-protecting group such as the trityl group.

The acylating acids of Formula Ill herein are either known compounds or are readily prepared by published procedures. European Patent Specification 7,470 published October 12, 1983 (application published February 6, 1980) exemplifies the preparation of compounds of Formula Ill wherein R2 is methyl, ethyl, propyl and isopropyl. U.S. Patent 4,390,534, referred to in the Prior 10 Art section, above, exemplifies the preparation of a wide variety of compounds of Formula Ill wherein R2 is, for example, cyclopenty], 2-eyclopenten-l-yi, allyi, 2- propynyl, 1-tert. butyloxycar bonyl-l-methylethyi, 1-tert. butyloxycarbonyi-l-cyclopenty], 1- ethoxycarbonyl-l-methylethyi, tert.

butyloxycarbonyimethyi, 1-tert. butyloxycarbony]-2-methylpropyi, trityl, and the like.

Compound 11 herein (7-amino-3-chloromethyl-3-cephem-4-carboxylate), used as a starting ma terial in Reaction Schemes la, 1b and lc, is a known compound.

The tertiary amines of Formula XI (and the secondary amines RR'NH) utilized in preparing the quaternary ammonio compounds of this invention are either known compounds or are readily prepared by those of ordinary skill in the art. Many of the amines are commercially available.

The present invention also provides a process for the preparation of compounds of the 20 formula h c -CONH S 11 11 25 A s,'N H (D H CHCH-CM2 H Q R1 HN DR2 wherein R' is hydrogen or a conventional amino-protecting group, R2 is hydrogen, a straight or 30 branched chain alkyl group containing from 1 to 4 carbon atoms, a cycloalkyl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula R1 4 R 5 4 R, 4 COOH -C- CH=CH-0 -C- c=- C-0 or 1 0 1 R5 in which R3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)aikoxy, and R4 and R' are each independently hydrogen, methyl or ethyl, or R4 and R5, taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 50 carbon atoms, and @H= 0 is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof, which process comprises reacting a compound of the formula 26 GB2194790A 26 (0) m -C - CONN S 11 -Fl 82HN'S/ N H,, 0 R21 0 H COOB1 C"CH CH21 wherin RT is the same as R2 or is a group of the formula COOB1 R4 1 or - c- coo 91 15 1 X RS in which X, R4 and R5 are as defined above, BI is a conventional carboxyl- protecting group, B2 is 20 hydrogen or a conventional amino-protecting group, Z is chloro, bromo or iodo, and m is zero or one, with a tertiary amine Q-N (or sequentially with a secondary amine RR'NH and a compound of the formula R"Z), and, if m is 1, reducing the sulfoxide by conventional means, and subse quently removing all blocking groups by conventional means.

The present invention also provides a process for the preparation of compounds of the 25 formula N C -CONH 11 11 (D 30 1 N N N C H=CH- CH2- N -= 0 OHN OR2 0 0OG wherin R' is hydrogen or a conventional amino-protecting group, R2 is hydrogen, a straight or 35 branched chain alkyl group containing from 1 to 4 carbon atoms, a cycloalkyl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula R4 1 RS C 4 COON -C- CH=CH-0 -C- - c=c-0 1 0 -C- COOH 1 R5 or in which R3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)aikoxy, and R4 and R5 are each independently hydrogen, methyl or ethyl, or R 4 and R5, taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 55 carbon atoms, and TH =_ 0 is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof, which process comprises acylating a compound of the formula 27 GB2194790A 27 H2W 7N S (D 0 ' CH=CHCH2-H-a coo 0 XXlI 5 with an acid of the formula \X 11 82HN-S/N N %,0R2 1 or with an acylating derivative of said acid, wherein RT is the same as R2 or is a group of the formula COOBI R4 1 1 20 or 1 1 -C-COOB 1 X RS 25, 25 in which X, R4 and R5 are as defined above, B' is a conventional carboxyl- protecting group and B3 is hydrogen or a conventional amino-protecting group.

The reactions are carried out in a non-aqueous organic solvent such as dimethyl suifoxide, hexamethylphosphoramide, methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile and the like, or mixtures of such solvents. The reactions are conveni ently carried out at a temperature of from about - 1 O'C to about + 50'C; we normally prefer to conduct the reactions at room temperature. During the quaternization step, at least one mole of the tertiary amine should be used per mole of Compound IX, XlX, XXIII or XXIV; we normally prefer to utilize from about 25% to 100% excess of the tertiary amine.

Carboxyi-protecting groups suitable for use as B' in the above reactions are well-known to those skilled in the art and include aralkyl groups such as benzyl, p- methoxybenzyl, p-nitrobenzyl and diphenyimethyl (benzhydryl); alkyl groups such as t-butyl; haloalkyl groups such as 2,2,2- ' trichloroethyl, and other carboxyl protecting groups described in the literature, e.g. in U.K. Patent 1,399,086. We prefer to utilize carboxy-protecting groups which are readily removed by treat- 40 ment with acid. Particularly preferred carboxyl-protecting groups are the benzhydryl and t-butyl moieties.

Amino-protecting groups suitable for use as B2 are also well-known in the art, and include the trityl group and acyl groups such as chloroacetyl, formyl and trichloroethoxycarbonyl. Amino protecting groups which are readily removed by treatment with acid, e.g. the trityl group, are 45 preferred.

When the cephalosporin nucleus is utilized in the form of the 1 -oxide (m= 1), the 1 -oxide is prepared by known procedures such as oxidation with m-chloroperbenzoic acid, peracetic acid, sodium tungstate, etc. The 1-oxide subsequently may be reduced by known procedures, e.g.

reduction of the corresponding alkoxysulfonium salt with iodide ion in an aqueous medium. The 50 alkoxysulfonium salt itself is readily prepared by treatment of the 1- oxide with, for example, acetyl chloride.

In another aspect, this invention relates to novel intermediates of the formula N c- CONN 5 55 \\ 11 XXVIlI H2H S/ m H\ OR2 OJ:5 ChCHCH2Z wherein Z is chloro, bromo, or iodo, R 2 is hydrogen, a straight or branched chain alkyl group containing from 1 to 4 carbon atoms, a cycloalkyl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula 28 GB2194790A 28 R4 4 COON -C- CH=CH-R3 -C-C=- C-0 or R C -C- CODH 1 R5 b in which R3 is hydrogen, (lower)aikyl or carboxyl, X is halogen, hydroxy or (lower)aikoxy, and R4 15 and R5 are each independently hydrogen, methyl or ethyl, or R4 and R5, taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and salts and esters thereof. Also included are compounds of Formula XXVIII in which the amino and/or carboxyl groups are protected by conventional amino- protecting or 20 carboxyl-protecting groups. In still another aspect, this invention relates to novel intermediates of the formula R23 R24 CH=N - 5 R25 0 CH=CHCH2Z p MIX C9OR22. 30 wherein R22 is hydrogen or a conventional carboxyl-protecting group, and R23, R24 and R25 are the same or different and are hydrogen, hydroxy, (lower)alkyl or (lower)aikoxy; or a salt, solvate, hydrate or ester thereof.

In still another aspect, this invention relates to novel intermediates of the formula N2N -N 5 (D C00e XXII 40 wherein -1)N=-Q is a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof. 45 As used herein, the terms acylamino and acyloxy refer to an acylated amino or acylated hydroxy group in which the acyl moiety is (lower)alkanoyl (e.g. formyi, acetyl, propionyl, butyryl, isobutyryl, isovaleryl, etc.), aroyl (e.g. benzoyl, etc.), (lower)aikenesuifonyl (e.g. mesyl, ethanesulfonyl, etc.) or aryisuifonyl (e.g benzenesuifony], tosyi, etc.). As used herein, the terms "(iower)aikyi" "(iower)aikoxy", "(iower)aikylthio" (or the like) means straight or branched chain alkyl, alkoxy, alkylthio (or the like) groups containing from 1 to 50 6 carbon atoms, inclusive. Similarly, the terms (lower)aikenyl and (lower)alkynyl mean alkenyl or alkynyl groups containing from 2 to 6 carbon atoms.

Example 1

H C-CONH - J WCH-CeN ' H H @ H2H 0 n- OCH3 CO6Ci-;/ c] l1A 7/E=1/1 CH3 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidoj-3-[3-(1methylpyrrolidinio)- 1-pro pen-l-yll-3-cephem-4-carboxylate (1-1A) To a solution of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido]- 65 29 GB2194790A 29 3-[3-(1-iodo-propen-l-yi)-3-cephem-4-carboxylate ([X-1) (Z/E=2/1, 150 mg, 0.21 mmole) in ethyl acetate (2 mi) was added a solution of 1methylpyrrolidine (36 mg, 0.42 mmole) in ethyl acetate (1 mi) in one portion with stirring. The mixture was stirred for 15 minutes and diluted with isopropyl ether (10 mi) to form a precipitate, which was collected by filtration. A mixture of the solid (130 mg), formic acid (1 mi) and concentrated HCl (0.1 mi) was stirred at room temperature. After 1 hour, the reaction mixture was concentrated under reduced pressure, diluted with water (20 m]) and filtered. The aqueous solution was passed through a reverse phase column (the packing of PrepPAK-500/C1, cartridge, 100 mi), eluting with water and 10% CH,OH. The desired fractions were collected, and concentrated in vacuo to a small volume and freeze-dried to give 13 mg (12%) of the title compound (1-1A) (Z/E=1/1), melting at >2WC 10 (dec.).

IR VKBr cm-1 3400, 1760, 1660, 1610.

max UV: APhosphate buffer (pH 7) nm (E"Y-) 236 (372), 288 (322).

max 1 Cm -H 20NIVIR: 6D20 2.31 (4H, rn,), 3.12 (3H, s, N-CH3), 3.6 (5H, P11M I:

rn, 2-H & N), 3.79 (1H, s, 2-H), 4.1 (2H, cl, 25 11- J=8, CH2N), 4.2 (3H, s, OCH3), 5.36 (11-1, d, J=4.5, 6-H), 5.95 (3H, m, 7- H & 3-CH=C", 6.66 (1/2H, d, J=10, 3-CH cis), 7.0 (1/2H, d, J=16, 3-CH trans).

Example 2

C- CON H 5 11 11 N N (D 0 -N N,/' H2NJS.' J CH: ' 'EH-CH2 OCH3 cooe E 1-113 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3pyridinio)- 1-propen- l-yll-3-ce- phem-4-carboxylate (]- 1B) A mixture of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido1-3- (3-iodo-l-propen-l-yi)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole), pyridine (158 mg, 2 mmoles) in dimethyisuifoxide (DIVIS0) (1 mi was stirred for 1 hour at room temperature. To the mixture was added ethyl acetate (20 mi) to precipitate a solid (620 mg), which was added to formic acid (6 mi) containing sodium bisulfite (60 mg). The mixture was stirred for 30 minutes at 45 4WC and concentrated to dryness. The residue was dissolved in H20 (40 mi) and some insolubles were removed. The aqueous solution was charged on a column of reverse phase (PrepPAK-500/C,, 100 mi) eluting with H20 (300 mi) and 5% aqueous CH,,OH (800 mi), and the eluate was monitored by uv (254 nm) and HPLC. The fractions (5% aqueous CH301-1) containing the desired product were combined, concentrated to a small volume and lyophilized to 50 yield 40 mg (8%) of the title compound (1- 113), melting at >20WC (dec.).

IR: VKBr em-' 3350, 1760, 1660, i610.

max UV: APhosphate buffer (pH 7) nm (E'%) 240 (352), 258 (366), max 'em 267 (279), 290 (469).

NMR: 3D20+MSO-d6 3.74 (2H, br-s, 2-H), 4.20 (3H, s, OCH,), PPM 5.92 (1H, cl, J=4.5, 7-H), 6.15 (1H, m, 3-CH=CM, 7.04 (1H, d, J=16, 3-CH trans), 8.2 (2H, rn, Py-H.3,5), 8.62 (1H, rn, Py-H4), 8.97 (2H, m, Py-H2,6).

GB2194790A 30 Example 3

C CONH N _'S"N NII I N 4L G H2 0 n-CH=CH-CH2- Nv473 OCH3 CgOE) I-IB Z/E=4/1 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3pyridinio -l-propen-l-yll-3-ce- 10 phem-4-carboxylate (/- 1B) The chloropropenyl compound, diphenyimethyl 7[2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-(3-chloro-1propen-l-y])-3-cephem-4-carboxylate (V111-1) (Z, 937 mg, 1.5 mmoles) was added to a stirred solution of pyridine (237 mg, 3 mmoles) in DMSO (3 mi) containing Nal (11 mg, 0.075 mmole). The mixture was allowed to stand overnight at room temperature in the dark. The mixture was diluted with ethyl acetate (30 mi) to separate the precipitate which was then collected by filtration, washed with ethyl acetate (10 mi) and dried to give 350 mg of the blocked product. The precipitate was treated with formic acid (3.4 m]) containing sodium bisulfite (34 mg) for 30 minutes at 4WC. After removal of the formic acid, the residue was purified by reverse phase column chromatography (packing of PrepPAK-500/C,8 20 cartridge, 100 mi) by eluting with 5% aqueous CH30H. The fractions containing the desired product were combined on the basis of HPLC analysis, evaporated under reduced pressure and lyophilized to give 41 mg (5.5%) of the title compound (1-1B) (Z/E=4/1). Mp. >20WC (dec. ).

IR: VCr cm-J 3300, 1760, 1660, 1600. 25 max UV: APhosphate buffer (pH 7) nm (E"16) 237 (386), 250 (377), max I cm 258 (369, 265 (347), 280 (311).

NIVIR: 6D20 3.45 & 3.76 (each 1 H, cl, J = 16, 2-H), 4.18(3H, s, OCH3), 5. 34 (3H, m, CH=CH-CH, & 6-H), 5.92 (1H, cl, J=4.5, 7-H), 6.58 (4/5H, d, J= 11, 3-CH cis), 7.03 (l/5H, d, J=16, 3-CH trans), 8.12 (2H, m, Py-1-1..J, 8.56 (1H, m, Py-Hj, 8.82 (2H, m, Py-H2.6), EXAMPLE 4

N C-CONH 5 11, N N \ (D N H2H /P.,CH=CHCHI-N 03 1-1 C E S-'O'NH2 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-(2amino-5-thiazolo[4,5 -clpyridinio)-l-propen-1-yl]-3-cephem-4-carboxylate (I-IQ A stirred solution of diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)- 2-methoxyiminoacetami50 dol-3-(3-iodo-l-propen-1-yl)-3-cephem-4- carboxylate (IX-1) (E isomer, 714 mg, 1 mmole), 2aminothiazolo[4,5- c]pyridine [prepared according to the procedure of T. Takahashi et a/., Pharm. Bull (Japan), 2, 34 (1954)] and dry DIVISO (1 ml) was kept for 1 hour at room temperature. To the reaction mixture was added ethyl acetate (20 ml) to give a yellow powder (710 mg). Formic acid (7 ml) and sodium bisulfite (70 mg) were added to the powder (700 mg), and the mixture 55 was stirred for 30 minutes at 40-45'C. After evaporation, the residue was triturated with H20 (40 ml). Insolubles were filtered off, and the filtrate was chromatographed over a reverse phase column (PrepPAK-500/C,,, 100 ml), with H20 and 10% CH,OH as eluant. The fractions containing the desired product were combined, and the solvent was removed under reduced pressure. Lyophilization gave the desired product (I-1Q as a colorless amorphous powder of the E isomer. 60 Yield 110 mg (19%). Mp. >200'C (dec. ).

IR VKBr cm-1 3300, 1760, 1660, 1630, 1600.

max 31 GB2194790A 31 uv: APhosphate buffer (pH 7) nm (E I'I'.) 245 (499), 285 (286).

max 1 Cm NMR: &MSO-d6+D20 3.86 (3H, s, OCH,), 4.98 (1H, d, J=4.5, PPM 6-H), 5.2 (2H, m, CH=CH-CH2), 5.57 (11H, rn, 3-CH=CM, 5.96 (1H, m, 7-1-1), 7.16 (11-1, cl, J = 16, 3-CH trans), 8.36 & 8.45 (each 1 H, d J=7, Py-H), 8.92 (1H, s, Py-H).

Example 5

1-1D H 11 C-CONH S H H N (E), 1\ N ChCH-CH2-H (CH313 M2NJS-" F 't.' OCH3 0 G 00 Z/E=1/1 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidoj-3-(3- trimethylammonio1-propen- 1- yl)-3-cephem-4-carboxylate fl- 1D) To a solution of diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido]- 3-(3-iodo-l-propen-1-yl)-3-cephem-4-carboxylate (IX-1) (Z/E=2/1, 490 mg, 0.68 mmole) in ethyl acetate (14 ml) was added a 0. 1 M trimethylamine solution in ether (13.6 ml) in one portion. The mixture was stirred for 10 minutes and evaporated to dryness, and the residue was triturated with ether (20 ml). The resulting solid (490 mg) was added to trifluoroacetic acid (0.2 25 ml) containing one drop of anisole. After 1.5 hours' stirring, the mixture was evaporated to dryness under reduced pressure and the residual oil was triturated with ether (20 ml). The resulting precipitate was collected by filtration and dissolved in H20 (20 ml). Some insolubles were removed, and the aqueous solution was eluted on a C18 reverse phase column (the packing of PrepPAK-500/Cl, cartridge, Waters' 30 ml) using water as eluant. Fractions containing the 30 desired compound were combined and concentrated to a small volume and Iyophilized to afford mg (9.2%) of the title compound (I-1D) (Z/E=1/1) as a colorless amorphous powder, melting at >150'C (dec.).

IR: VKBr cm- 1 3300, 1770, 1670, 1605. 35 max uv: A Phosphate buffer (pH 7) nm (E 1%) 236 (389), 287 (343).

max 1 em NIVIR: &O 3.45 & 3.7 (1 H, d, J = 16, 2-H), 3.81 (1 H, s, 2-H), PPM @ 4.1 (21-1, d, J=8, -CH2N), 4.21 (31-1, s, OCH3), 5-39 (11-1, d, J=4.5, 6- H), 5.95 (21-1, m, 3-CH=CH & 7-H), 6.61 (1/21-1, d, J= 11, 3-CH cis), 7. 05 (1/21-1, d, J=16, 3-CH trans).

Example 6

H C- CONH - 5 NH2 j, 11 11 - N ED) D N2N S,, N N N CH=CH C H2\/17 \OCH3 E) coo 1-1E E 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3- aminopyridinio)1-propen- 1 yll-3-cephem-4-carboxylate (1-1E) Diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2- methoxyiminoacetamido]-3-(3-iodo-1 -pro- 60 pen- 1 -yl)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) was added to a stirred solution of 3-aminopyridine (188 mg, 2 mmoles) in DIMS0 (1 ml). The mixture was stirred for 1 hour and diluted with ethyl acetate (20 mi). The resulting precipitate was collected by filtration, washed with ethyl acetate and dried to give 520 mg of yellow powder. A mixture of the powder (500 mg), formic acid (5 mi) and sodium bisulfite (50 mg) was stirred for 30 minutes at 40'C. The 32 GB2194790A mixture was concentrated in vacuo, dissolved in H20 (40 ml) and filtered to remove insolubles. The aqueous solution was chromatographed on a column of reverse phase (packing of PrepPAK-500/C18, 100 mi), with 7.5% aqueous CH30H elution. The fractions containing the desired compound were evaporated and lyophilized to give the title compound (1-1Q (7 mg, 1.4%), 5 melting at >l8WC (dec.).

IR VKBr cm-1 3400, 1765, 1675, 1620, 1600.

max UV: APhosphate buffer (pH 7) nm (E "Y-) 246 (403), 290 (468).

max I Cm NMR: 3DO 3.72 (2H, m, 2-H), 4.14 (3H, s, OCH3), 5.35 (3H, m, P6-H & CH=CH-CH2), 5.9 (1H, d, J=4.5, 7-H), 6.1 (1H, rn, 3-CH=CM, 7.05 (1H, d, J=16, 3-CH, trans), 8.1 (1H, m, Py-HJ, 8.54 (1H, br-s, Py-H6), 8. 68 (1H, m, Py-H4), 9.4 (1H, rn, Py-H2).

Treatment of IX-1 (716 mg, 1 mmole) with 3-t-butoxycarbonylaminopyridine (324 mg, 2 mmoles) by a procedure similar to that described above gave 12 mg (2.3%) of 1-1E.

Example 7

N 7-1- CONH T N 5 - 11 E) N 0 N IF "'OCH3 0 0 M2N n[H CHCH2 ú00 1-1E Z/E=1/1 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3- aminoI-pyridinio)- 1-pro pen-1-yl]-3-cephem-4-carboxylate (I-1E) A mixture of diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3- (3-iodo-l-propen-1-yl)-3-cepthem-4-carboxylate ([X-1) (Z/E=2/1, 500 mg, 0. 7 mmole) and 3 aminopyridine (66 mg, 0.7 mmole) in dimethylsulfoxide (1 ml) was stirred for 20 minutes at 35 room temperature. The mixture was diluted with ethyl acetate (10 ml) and ether (10 ml), and the resulting precipitate was collected by filtration, washed with ether (10 ml) and dried. The quaternized salt was dissolved in formic acid (3 ml) containing concentrated HCI (0.3 ml) and stirred for 1.5 hours at room temperature. The mixture was concentrated to dryness under reduced pressure. The residue was dissolved in 2% HCI (10 ml) and filtered. The aqueous layer 40 was chromatographed on a reverse phase column (PrepPAK-500/C,, 100 ml). After washing with water (500 ml), the column was eluted with 5% aqueous CH,OH. The fractions containing the title compound were combined, concentrated in vacuo and freeze-dried to give 15 mg (4.2%) of the title compound (I-1E) (Z/E=1/1) as a colorless amorphous powder. Mp. >160'C (dec.).

IR: VK1k cm-1 3400, 1765, 1675, 1620, 1600.

max UV: APhosphate buffer (pH 7) nm (E'%) 244 (434), 287 (333).

max 1 CM NIVIR: &MSO-de+D20 3.73 (2H, m), 4.14 (3H, s, OCH,), 5.35 (3H, PPM m, 6-H & CH=CH-CH2), 6.0 (2H, m, 7-H & 3-CH=CM, 6.6 (l/2H, d, J= 11, 3-CH cis), 7.05 (l/2H, d, J=16, 3-CH trans), 8.08 (1H, m, Py-HJ, 8.6 (2H, rn, Py-H4A, 9.4 (1 H, rn, Py-H2).

33 GB2194790A 33 Example 8

N N, N2Nk S,-' OCH3 0 100 0 CW-CHCH2- 11, 11F E 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3formylaminopyridinio)- 1 -pro-10 pen-l-yll-3-cephem-4-carboxylate (1-1F) A mixture of diphenyimethyl 7-[2(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamido1-3(3-iodo-lpropen-l-yi)-3-cepthem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) and 3formyiaminopyridine [prepared according to the procedure of N. Enomoto et al., Bull, Chem. Soc. Japan, 45, 2665 (1972)l (244 mg, 2 mmoles) in DIVISO (2 mi) was stirred at a room temperature for 1 15 hour, and poured into ethyl acetate (200 mi). The precipitate was collected by filtration, washed well with ethyl acetate and dried. A mixture of the quaternized salt (500 mg) and sodium bisulfite'(50 mg) in HCOOH (5 mi) was stirred at 40-50'C for 80 minutes and evaporated to dryness in vacuo. The residue was dissolved in water (40 mi), neutralized with NaHC03 and then filtered to remove insoluble material. The clear filtrate was chromatographed on a reverse phase 20 column, PrepPAK-500/C18 (100 mi), with water and 5% CH30H, 10% CH30H, 20% CH30H and 30% CH30H. The fractions containing the desired compound were combined, concentrated in vacuo and lyophilized to give 16 mg (2.9%) of the title compound (1-1F) (E) as a tan powder. Mp.>l7WC (dec.).

IR UV:

VKBr cm-1 3340(br), 1760, 1670, 1620(br), 1590.

max 1Phosphate buffer (pH 7) nm (P'/) 218 (428), 248 (362), Max 1 CM290 (474).

NIVIR: 3D20 + NaHC03 PPM 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH,), 5.91 (1H, d, J=4.5, 7-H), 6.25 (1H, m, CH=CH-CH2), 6.98 (1H, d, J=16, 3-CH trans), 8.8-7.9 (4H, m, Py-H), 9.38 (1H, br, NHCHO).

Example 9

N C CONH S 11 1 h, + 2TNS - li 40 N2N j, S.-' N N 1.1 QICH3 N Q- CH=CHCH2 1-1G NH2 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3carbamoylpyridinio)- 1-pro pen- l-yll-3cephem-4-carboxylate (1-1G) To a solution of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamidol- 3-(3-iodo-l-propen-l-yi)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) in DIVISO (2 mi) was added nicotinamide (244 mg, 2 mmoles), and the mixture was stirred at ambient tempera- 50 ture for 1.5 hours and poured into ethyl acetate (200 mi) with stirring. The resulting precipitate was collected by filtration. The quaternized salt (500 mg) was dissolved in HCOOH (5 mi) in the presence of sodium bisulfite (50 mg), and the mixture was heated at 40- 50T for 40 minutes, with stirring, and evaporated to dryness. The residue was dissolved in water (40 m]) and an insoluble solid was filtered off and washed with a small amount of water. The filtrate and wash 55 were combined and chromatographed on a reverse phase column, PrepPAK-500/C, (100 mi). After elution with water and 5%, 10% and 20% aqueous CH30H, successively, the fractions containing the desired material were combined, concentrated in vacuo and freeze-dried to yield 21 mg (3.8%) of the title compound (1-1G) (E) as a yellow powder. Mp. >175'C (dec.).

IR 0111 em-' 3340 (br), 1760, 1670, 1600.

wax 34 GB2194790A 34 IJIV: APhosphate buffer (pH7) nm (E"".) 235 (326), 274 (sh, Max NMR: 5D20+NaHC03 PPM 1 405), 290 (446).

3.68 (2H, br, 2-H), 4.15 (3H, s, OCH3), 5.32 (1 H, cl, J = 4.5, 6-H), 5. 45 (1 H, d, J = 7, CH=CH-CH,), 5.88 (1H, d, J=4.5, 7-H), 6.15 (1H, d-t, J=16 & 7, 3-CH=CI-4, 7.00 (1H, d, J=16, 3-CH trans), 8.23 (1H, m, Py-H,), 9.03 (2H, m, Py-H4&6), 9.34 (1H, s, Py-H2), Example 10

N __ f - CONN S I I I A (DCON H US,,,N' N FN 'A N 'n H=CHCH2-N 2 \OCM3 cool, ' 1-1H E 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4carbamoylpyridinio)- 1 -pro- 20 pen-l-yll-3-cephem-4-carboxylate (1-114) To a stirred solution of diphenyImethyl 7-[2-(5-amino-1,2,4-thiadiazol-3yi)-2-methoxyiminoacetamidol-3-(3-iodo-l-propen-l-yl)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) in dry DIVISO (2 mi) was added isonicotinamide (244 mg, 2 mmoles). The mixture was stirred at room temperature for 1 hour and then poured into ethyl acetate (200 mi). The resulting precipitate 25 was collected by filtration, washed well with ethyl acetate and dried. A stirred mixture of the quaternized material (40 mg) and sodium bisulfite (40 mg) in HCOOH (4 m]) was heated at 40-50'C for 1 hour, and evaporated to dryness under reduced pressure. The crude solid was dissolved in water (40 mi). After filtration of an insoluble material, the filtrate was chromato graphed on a reverse phase column (packing of PrepPM/C1, 100 mi) using water and 5%, 10%, 20% and 30% aqueous CH30H as eluant. The fractions containing the desired compound were combined, evaporated and lyophilized to give 21 mg (3.8%) of the title compound (1-1H) (E) as a pale yellow powder. Mp. >l8WC (dec.).

IR VKBr cm-1 3340(br), 1760, 1670, 1600.

max uv: APhosphate buffer fpH 7) nm (E'%) 222 (362), 285 (452), Max 1 CM NMR: 6D20 N,HC03 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH3), 5.33 40 PPM (11-1, d, J=4.5, 6-H), 5,46 (2H, d, J=7, CH=CH-CH2), 5.90 (11-1, d, J=4.5, 7-H), 6.17 (11-1, d-t, J16 & 7, 3-CH=C", 7.02 (11-1, d, J= 16, 3-CH trans), 8.43 & 9.09 (each 2H, d, 45 J=7, Py-H).

Example 11

E- CONH NI HN H 'ACH3 0 H 0 CH=CHCH2-N 2 coo 1-11 E CH2NH2 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3aminomethylpyridinio)- 1- propen-l-yll-3,cephem-4-carboxylate (1-11) A mixture of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido1-3- (3-iodo-l-propen-l-yi)-3-cephem-4-carboxylate (]X-1) (E, 716 mg, 1 mmole) and 3-(t-butyloxycarbonylaminomethyl)pyridine (516 mg, 2 mmoles) in DIVISO (2 mi) was stirred at ambient tempera ture for 30 minutes. The mixture was poured into ethyl acetate (200 mi), and the precipitate was collected by filtration, washed well with ethyl acetate and dried. A mixture of the quater nized salt (500 mg), sodium bisulfite (50 mg) in HCOOH (5 mi) was stirred at 40-50C for.80 minutes and evaporated to dryness under reduced pressure. The residual solid was dissolved in GB2194790A 35 water (40 mi), and the mixture was neutralized with Nal---1CO, Insoluble material was filtered off, and the filtrate was chromatographed on a reverse phase column (packing of PrepPAK-500/C, cartridge, 100 mi), eluting with water, 5%, 10%, 20% and 30% aqueous CH,OH, successively. The fractions containing the desired compound were combined, evaporated and lyophilized to 5 provide 10 mg (1.8%) of the title compound (1-11) (E) as a tan powder.

IR VKBr cm-1 3380(br), 1760, 1650(sh), 1620(sh).

max uv: ZPhosphate buffer (pH 7 nm (E'%) 235 (sh, 260), 286 maX 1 em (370).

NMR: 6D20+NaHCO3 3.68 (2H, br, 2-H), 4.16 (3H, s, OCH3), 6.98 ppm (1 H, d, J = 16, 3-CH trans), 8.05 (1 H, m, Py-H5), 8.50 (1 H, m, Py-H4), 8.80 (2H, m, Py-H2,6), Example 12

N CONH - N H OCH CK02 "N CONN2 o:N 3 COOG 1-1H E 25 7-[2-(5-Amino1,2,4-thiadiazol3-yl)-2-methoxyiminoacetamidol-3-[3-(4carbamoylpyridinio)- 1-pro pen- l-yl)-3-cephem-4-carboxylate (1-1H) A mixture of diphenyImethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido1-3- (3-iodo-l-propen-l-yi)-3-cephem-4-carboxylate (]X-1) (E isomer, 4.1 9, 5. 7 mmoles) and isonico- 30 tinamide (1.4 9, 11 mmoles) in dry DMSO (6 ml) was stirred for 2 hours at room temperature while monitoring by TLC (silica gel plate, CHC13:C11301---1=3:1). The reaction mixture was diluted with ethyl acetate (100 mi) to separate a yellow gum, which was treated with formic acid (40 m]) and sodium bisulfite (390 mg) at 45'C for 30 minutes. The resulting solution was concen trated to dryness. The residue was dissolved in H20 (100 mi) and insolubles were removed by 35 filtration. The combined filtrate and water wash was applied to the top of a column containing reverse phase packing (PrepPAK/500/C,, 120 m]). The column was eluted with H20. The eluate was collected in 300 mI fractions and monitored by uv (254 nm) and HPLC (Lichrosorb RP-18, 4 x 300 mm, 0.0 1 M ammonium phosphate buffer, pH 7.2 containing 20% CH30H). Fraction Nos. 4 and 5 were combined and concentrated to a small volume. Lyophilization gave 250 mg 40 (8.1%) of the title compound 1-1H, melting at >l8WC (dec.).

The spectra indicated that the product was identical to that obtained in Example 10.

Preparation of the hydrochloride-To a suspension of Compound 1-1H (98 mg, 0.18 mmole) in CH30H (1 mi) was added 10% HCI (0. 1 mi), and the mixture was stirred for 5 minutes. To the resulting yellow solution was added acetone (100 mi) to give a precipitate, which was collected 45 by filtration, washed with acetone (2 x 10 mi) and dried in vacuo to give the hydrochloride salt of 1-1H as a coiorless powder. Yield 88 mg (79%). Mp. >1WC (dec.).

IR j,KBr cm-1 3300, 1770, 1680, 1620.

max uv: APhosphate buffer (pH 7) nm (E 1%) 227 (385), 286 max 1 cm(374).

NMR: JD20 2.32 (1H, s, acetone-H), 3.79 (2H, br-s, 2-H), 4.17 Pp. 3H, s, OCH3), 5.34 (1H, d, J=4.5, 6-H), 5.49 (2H, d, J=7, CH=CH-CH2), 5.93 (1H, d, J=4.5, 7-H), 6.28 (1H, d-t, J=16 & 7, 3-CH=CM, 7.15 (111, d, J=16, 3-CR, 8.43 & 9.1 (each 2H, d, J=7, Py-H).

0 36 GB2194790A 36 Example 13

S N-W-c.-CONN ) H H EIN coo I-ii E 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3[3-(2methyi thiazolio)-1-propen-l- 10 yll-3-cephem-4-carboxylate (/- W) To a mixture of diphenyimethyl 7-[2-(5amino-1.2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol3-(3-iodo-l-propen-lyl)-3-cephem-4-carboxylate (IX-1) (E, 714 mg, 1 mmole) and 2methylthiazoie [prepared according to the procedure of R. P. Kurkjy, E. V. Brown, J. Am. Chem. Soc., 74, 5778 (1952)] (198 mg, 2 mmoles) in dry CHCl, (10 mi) was added AgBIF, (90% pure, 217 mg, 1 mmole) at -20'C. The mixture was stirred for 30 minutes at room temperature and filtered. The precipitate was extracted with 10% CH,01-1-CHC13 (3x20 m[). The combined extracts were washed with brine (2x5 mi), dried over M9S04 and evaporated to dryness to give a yellow residue, which was triturated with isopropyl ether and filtered to yield 350 mg of the quaternized product. A mixture of this solid, sodium bisulfite (35 mg) and formic acid (3.5 mi) was stirred at 20 400C for 30 minutes. The mixture was concentrated to remove the formic acid, and the residue was diluted with H20 (40 m]). Some insolubles were removed by filtration. The filtrate was placed on a reverse phase column (PrepPAK-500/C1, 100 mi). The column was eluted with H20 (200 mi), 5% aqueous C1-1,0H (400 m]) and 10% aqueous CH30H (300 mi), successively.

The fractions containing the desired product was pooled on the basis of HPLC analysis (Lichro- 25 sorb RP-18, 4x300 mm, 0.01 M ammonium phosphate buffer pH 7.2, containing 20% CH30H).

The combined solution was concentrated to a small volume and lyophilized to give 40 mg (7.7%) of the title compound (1-1J) (E). Mp. >l9WC (dec.).

IR VKBr cm--I 3300, 1760, 1660, 1600.

max uv: APhosphate buffer (pH 7) nm (E'%) 238 (442), 292 (421).

max 1 cm NIVIR: &20DMSO-d, 3.06 (31-1, s, thiazole-CH3), 3.74 (2H, br-s, PPM 2-1-1), 4.19 (31-1, s, OCH,), 5.92 (11-1, d, J=4.5, 7-1-1), 6.1 (11-1, m, 3-CH=C", 6.8 (1 H, d, J = 16, 3-CH trans), 8.04 & 8.23 (each 11-1, d, J=4, thiazole-H.

Example 14

N C-CONN 5 11 11 N N N EH =EH EH) NF-"\CHH W2J,,5, 4 OCH3 p n- 2 CODE) \==Y 1-1L E 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4hydroxymethylpyridin io)- 1 propen-l-yll-3-cephem-4-carboxylate (1-1Q A mixture of diphenyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-(3-iodo- 1-propen-l-yi)-3-cephem-4-carboxylate (IX-1) (E isomer, 1.07 9, 1.5 mmole), 4-hydroxymethyl pyridine (818 rng, 7.5 mmole) in CH3CN (4.5 mi) and CH30H (3 mi) was stirred at room temperature under N2 atmosphere for one hour. After removal of the solvents by evaporation, the residual oil was triturated with isopropyl ether, collected by filtration, and washed with a mixture of isopropyl ether and methanol (3A, 10 mi) to give 1.28 9 of the quaternized cephem ester as a yellow powder. A solution of the quaternized ester (1.25 9) and sodium bisulfite (600 mg) in 85% HCOOH (10 mi) was stirred at room temperature under N2 atmosphere for one hour. 60 After the addition of 85% HCOOH (5 mi), the mixture was stirred under the same conditions for an additional hour. Toluene was added and the reaction mixture was evaporated azeotropically under reduced pressure. The residue was triturated with acetone to yield 1.17 9 of the crude formate of the title compound. A suspension of this compound (1.15 9) in water (100 m]) was filtered to remove insolubles, which were washed with water (10 mix2). The filtrate and the 65 37 GB2194790A 37 washes were combind and subjected to reverse phase column chromatography. The column, which was packed with the packing taken out of a prepPAK- 5001C,, cartridge column (Waters) 60 mi), was developed with water, 5% methanol and 10% methanol, successively. The fractions containing the desired compound were combined, concentrated under reduced pressure, and precipitated by the addition of acetone to give 100 mg of the title compound (1-1L) as a pale yellow powder. To a suspension of the powder (90 mg) in methanol (9 mi) was added 1 M HCl in CH.0H (0.5 mi) and the mixture was stirred at room temperature and concentrated in vacuo. To the concentrate was added isopropanol to precipitate 77 mg of hydrochloride of the title compound. Pale yellow powder. M.p. >190oC (dec.).

max IR: v',", cm-1, 1775, 1670, 1635, 1530.

max ILIV: APhosphate buffer (pH 7) nrn (e), 230 (22600), 264 (sh, 16300) JD2 0 3.83 (2H, br. 2-CH), 4.17 (3H, s, ppm OCH3), 5.06 (2H, s, CH20H), 5.36 0H, cl, 20 J=4.5 Hz, 6-H), 5.41 (2H, d, J=7 Hz, CH=CH-CH2), 5.94 (1H, d, J=4.5 Hz. 7-H), 6.36 (1H, d-t, J=16 and 7Hz, CH=CHCH2), 7.13 (1H, d, J16 Hz, CH=CH-Cl-1J, 8.08 and 8.83 (each 2H, d, 25 J=7 Hz, Py-H).

Example 15 -CONH ',' 5 30 1---1 H N IM\ N2N 5--- 01- n WCHCH2- U \-CON H 2 \ 0C2NS COOO "IC=F 1-2H E 35 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamidol-3-[3-(4carbamoylpyridinio)- lpropen l-yll-3-cephem-4-carboxylate (1-2H) To a solution of 200 mg of 7-amino-3-[3-(4-carbamoylpyridinio)-1-propen-lyl]-3-cephem-4- carboxylate hydrochloride (E isomer) in 5 mI of 50% aqueous acetone was added portionwise 40 mg of 2-ethoxyimino-2-(5-amino-1,2,4-thiadiazol-3-yi)acetyl chloride hydrochloride [prepared according to the procedure described in published Japan patent application (Kokai) 57-24389 (2/8/82)], and the niixture was adjusted to pH 6.5-7.0 with 2 N Na2C03 (about 1 mi). The reaction mixture was stirred at 10'C for an hour, acidified to pH 2 with 1 N HCl and evaporated in vacuo. The residqe was filtered and the filtrate was chromatographed on a column of HP-20, 45 which was eluted with 500 mI of water and 25% aqueous isopropanol, successively. Fractions containing the desired product were combined and evaporated under reduced pressure. The oily residue was treated with isopropanol (20 m]) to give 263 mg (93%) of the title compound (1-2H). M.p. 170'C (dec.).

To a stirred suspension of 225 mg (0.40 mmole) of the above zwitterion in 10 mi of methanol was added 1 mi of 1 N HCl in CH30H and the mixture was stirred at room temperature for 30 minutes. The solution was filtered and concentrated under reduced pressure. To the residue was added 15 mI of isopropyl alcohol, and the resulting precipitate was collected by filtration and dried in vacuo to give the title compound as its hydrochloride. Yield 146 mg 55 (57%). M.p. 1600C (dec.). Estimated purifty 65%.

M VKBr cm-1, 3300, 1780, 1680, 1620.

max UV: APhosphate buffer (pH 7) nm (e), 227, (22300), 288 (22800).

max 38 GB2194790A 38 NIVIR: 6D20 1.44 (31-1, t, J=7 Hz, OCH2-CH3), pp3.74 (2H, br. s, 2-H) 4. 45 (2H, q, J=7 Hz, OCH27CH,), 5.36 (1H, d, J=4.5 Hz, 6-H), 5.46 (2H, d, J=7 Hz, 3-CH=CH-CH2), 5.92 (11-1, d, J=4.5 Hz, 7-H), 6.20 (1H, m, 3-CH=CM, 7.04 (11-1, d, J=16 Hz, 3-CH=CH), 8.43 (2H, d, J=7 Hz, Py-HA), 9.10 (2H, d, J=7 Hz, Py-1-1j.

Example 16

7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4carbamoylpyridinio)- 1 -pro- 10 pen- l-yll-3-cephem-4-carboxylate (l- M) (E isomer) This Example shows the preparation of Compound 1-1H via the last few steps of Reaction Scheme la or lb, wherein the intermediate benzhydryl 7-[2-(5-amino-1,2,4thiadiazol-3-yi)-2methoxyiminoacetamidol-3-[3-(4-carbamoylpyridinio)-1propen-l-yil-3-cephem-4 -carboxylate for- mate (XXVII-1ffi is isolated.

A. Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-. 2methoxyiminoacetamidol-3-[3-(4-carbamoyl1-pyridinio)-1-propen-l-yll-3-cephem-4-carboxylate Formate (E isomer) (XXVII-M) A solution of X11-1H (Y0=10, E isomer) (34 9, 75% pure) in a mixture of acetone and CH30H (l/1, 200 m[) was placed on a column of Amberlite IRA-410 (formate form 340 mi). The column was eluted with the same solvent system. The first fraction (1 L) was evaporated to about 100 mi of the volume and the brown residue was triturated with isopropyi ether (400 m[). The resulting powder was collected by filtration and dried under vacuum to afford 29 g (75% pure by HPLC) of the title compound XXVII-11-1 (E isomer) as a brown powder melting at >l5WC (dec.).

IR VK13r cm-1 3300, 1780, 1680, 1630, 1600.

max 20' UV: AEtOH nm (E'%) 282 (186). 30 max 1 CM NIVIR: jacetone-dICH3OHd(1 11) 4.0 (31-1, s, OCH,), 5.26, (1 H, d, PPM J=4.5 Hz, 6-H), 5.43 (2H, d, J=7 Hz, CH2W), 5.99 (1H, d, J=4.5 Hz, 7-H), 6.5 (11-1, rn, 35 3-CH=CM, 6.92 (11-1, s, ChPh2), 7.1 (11-1, d J=16 Hz, 3-CM, 7.35 (10H, m, Ph-M, 8.3 (1 H, s, HCOO), 8.46 & 9.12 (21-1 each, d, J=8 Hz, Py-H).

40 B.7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4- carbamoyl 1-pyridinio)- 1-propen-l-yll-3-cephem-4-carboxylate (I-IH) (E isomer) A mixture of MVII-1H (E isomer) from Step A (29 9, 75% pure) and 85% formic acid (290 mi) was stirred for 2 hours at room temperature. Evaporation of the mixture gave a brown oil which was triturated with acetone (500 mi). The powder was collected by filtration, washed 45 with acetone (2 X 100 m]) and drid in vacuo to give 24 9 (50% pure by HPLC) of the crude title compound. The brown solid was treated twice with 2 N HCI (1 L and 0.5 L). The aqueous extracts were combined and placed on a column packed with Diaion HP-20 (1. 5 L). The column was washed with water (8 L) and eluted with 30% CH,OH (5 L). The fraction containing the desired product was evaporated to about 30 mi. The concentrate was treated with acetone (200 50 m]) to give a precipitate, which was collected by filtration and dried in vacuo to give 10. 1 g (85% pure) of the title compound (zwitterion form) as a yellow powder. To a suspension of this product in CH30H (100 mi) was added N HCI in CH30H (55 mi) at room temperature and the mixture was stirred for 30 minutes. The resulting clear solution was filtered to remove insol ubles, concentrated to about 50 mi of the volume and precipitated with isopropanol (200 mi). 55 The resulting powder was collected, washed with isopropanol (50 mi) and dried in vacuo to give 10.5 g (85% pure) of the title compound 1-11-1 (E isomer) (HCI salt), melting at >l8WC (dec.).

Pale yellow powder.

Example 17 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol3-[3-(4-carbamoylpyridinio)- 1 pen- l-y11-3-cephern-4-carboxylate (1- M) (E isomer) This example shows the preparation of Compound 1-1H via the last few steps of Reaction Scheme la or lb, wherein intermediate MVII-1H (the formate) is not isolated.

A solution of IX-1 (E isomer) (27.6 9, 38.5 mmole) and isonicotinamide (22.8 9, 187 mmole) 65 39 GB2194790A 39 in a mixture of CH3M (120 mi) and CH,OH (100 mi) was stirred at room temperature for 1 hour under a nitrogen atmosphere. After evaporation of the organic solvents, the oily residue was triturated with isopropyl ether to give 50.5 9 of a mixture of the quaternized salt and isonicotinamide. A solution of the mixture (50.3 9) and sodium bisuifite (16 9) in 85% HCOOH (160 m]) was stirred at room temperature for 40 minutes and subsequently at 40'C for 1 hour under N, The mixture was evaporated in vacuo. The residual oil was mixed with toluene (50 m]), evaporated azeotropically and triturated with acetone (400 m[) to give 27. 8 9 of the crude title compound. This material was treated twice with 2 N 1HIC1 (1 L and 0.5 L). The acid extracts were combined and placed on a column of HP-20 resin (1.5 L). The column was eluted with water (90 and 30% methanol OOL). The fractions containing the desired compound were combined and concentrated to give a yellow oil, which was triturated with acetone (300 mi) to yield 9.35 g of the zwitterion form of the title compound.

To a suspension of the product (9.3 g) in CH,OH (180 mi) was added 1 N HCl in CH30H (55 mi) to obtain a clear solution. The solution was concentrated to about 100 m] and diluted with isopropanol to precipitate 9.50 9 (purity 75%) of the title compound 1- 11H (IE isomer) as its 15 hydrochloride. Pale yellow amorphous powder. M.p. >195'C (dec.).

Example 18 7-[2-(5Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol3-[3-(4-carbamoylpyridinio)- 1-pro- pen- l-yll-3-cephem-4-carboxylate (lM) (E isomer) This example shows the preparation of Compound 1-1H via the last step (7- N-acylation) of Reaction Scheme le.

To an ice-chilled suspension of the 7-amino-cephem hydrochloride XM-H (E isomer) (5.0 g, 12.6 mmole) in 50% aqueous acetone (100 mi) was added sodium bicarbonate in small portions. The pH of the mixture was monitored by a pH meter throughout the reaction. To the cold neutralized solution (pH about 7) was added 2(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetyl chloride hydrochloride (4.02 9, 15.6 mmole) in small portions over a period of an hour, and the pH of the reaction mixture was maintained in the range of 6.8-7.5 by occasional additions of sodium bicarbonate. The reaction was also monitored by tic. After all of Compound XXII-H had been consumed, the mixture was acidified to pH 3 by the addition of 2 N hydrochloric acid. 30 The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was diluted with acetone (400 m]) to separate the precipitate, which was collected by filtration to afford 9.59 9 of the crude title compound as a light yellow powder. Estimated purity 40% by HPLC. A suspension of the crude product (9.5 g) in 2 N hydrochloric acid (150 mi) was filtered, - and the filtrate was adsorbed on a column of HP-20 resin (500 mi). After washing with water 35 (1.5 L), the column was eluted with 25% aqueous isopropyl alcohol and the eluate was collected in 100-mi fractions. The desired fractions were pooled, acidified with 2 N hydrochloric acid (10 mi) and concentrated. The residual oil was triturated with isopropyl alcohol (200 mi), and the precipitate was collected by filtration. After drying over phosphorus pentoxide, 5.18 g of the title compound 1-1H (IE isomer) hydrochloride was obtained as a yellow amorphous powder. M.p. >l90'C (dec.). Estimated purity 75%.

Example 19

Purification and crystallization of Compound 1-1H (E isomer) Compound 1-1H hydrochloride obtained in Example 16 was a pale yellow amorphous powder 45 of 85% purity.

Procedure 1 Six grams of the 85% purity hydrochloride was dissolved in 20 m] of H20 and filtered through a celite pad. The ambercolored filtrate (pH 2) was passed through a reverse phase column (the 50 packing of prepPAK-500/C18 cartridge, Waters; 120 m]), which was eluted with water. The eluate was collected in 120-mi fractions with monitoring by HPLC. Fraction No. 3 through fraction No. 5 were combined and concentrated to about 10 m], and precipitated by acetone (100 m]) to give 3.3 9 of the zwitterion form of 1-111 (pale yellow amorphous powder; estimated purity 95%).

To a suspension of the 95% purity powder (3.2 9) in CH30H (32 mi) was added N HO in CH30H (18 mi), and the mixture was stirred at room temperature until a clear solution was obtained. The solution was filtered and the filtrate was concentrated to about 10 mi. To the concentrate was added isopropanol (100 mi) to separate a pale yellow precipitate, which was collected by filtration, washed with isopropanol (5 mi) and dried to yield 2.6 g of the HCI salt 60 (amorphous powder; estimated purity 95%).

A solution of the 95% purity hydrochloride (1 g) in water (4 mi) was adjusted to pH 6.5 with NaHC03 (200 mg) and stirred for 30 minutes. The crystals which separated during stirring were collected by filtration, washed with water (2 x 5 ml) and drid in vacuo to give 7 10 mg of l- 1 H (zwitterion form) as pale yellow prisms. M.p. >185'C (dec.). Microanalysis showed it to be the 65 GB2194790A 40 trihydrate.

IR VKBr cm-1 1780, 1695, 1660 1630, 1610.

max UV: 'I Phosphate buffer tpH 7) nm (e) 227 (22000), 290 (23000).

max Anal. Calc'd for C.^^O,S2. 3H30. C, 42.14; H, 4.38; N, 18.72; S, 10.71.

Found: C, 42.4 1; H, 4.35; N, 18.86; S, 11.00.

NMR: JDMSO-d6+D,0 3.45 (2H, br, s, 2-H), 3.9 (3H, s, OCH3), PPM 4.99 (1H, d, J=4.5 Hz, 6-H), 5.16 (2H, d, 10 J=7 Hz, CH2W), 5.61 (1H, d, J=4.5 Hz,7-H), 5.8 (1H, d-t, J=16 & 7 Hz, 3-CH=CP4, 6.93 (1 H, d, J=16 Hz, 3-CM, 8.18 & 8.89 (each 2H, d, J=7 Hz, py-H).

15 20.

Procedure 2 Once crystalline 1-1H had been obtained from Procedure 1, it was possible to obtain the crystalline zwitterion form of 1-11-1 directly from the crude 1-11-1 hydrochloride by seeding with a few crystals of the pure 1-1H.

A solution of the 85% pure hydrochloride (250 mg) in water (1 mi) was treated with charcoal.

The solution was adjusted to pH 6.5 with NaHC03 (60 mg) and decolorized with charcoal. The 25 filtrate was seeded with a few pieces of the crystals obtained from Procedure 1 and stirred overnight at room temperature. The separated crystals were collected by filtration, washed with water (2x2 mi) and dried under reduced pressure to give 170 mg (80% recovery) of pale yellow prisms of 1-1 H (zwitterion form), melting at > 185'C (dec.), which was identical with that obtained by Procedure 1, (as shown by IR, UV, NMR).

The crystalline zwitterion form of Compound I-IH was slightly soluble in water (6 mg/mi in saline at 23'C).

Example 20

N C - CONN - 5 CH20H C2CH CH.) N H 2N"l"S"'N N'9CH3 Op?0 - 2 coo 1-1K E 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3hydroxymethylpyridin io)- 1 propenyll-3-cephem-4-carboxylate (1-1K) (E isomer) A. Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-[3-(3-hydrox ymethylpyridinio)-1-propenyll-3-cephem-4-carboxylate iodide (E-isomer) (X11-1K) To a solution of IX-1 (E-isomer, 1.79 9, 2.5 mmoles) in 2.5 mi of CH,OH and 7.5 mi of CH,CN was added 3-hydroxymethylpyridine (545 mg, 5 mmoles), and the mixture was stirred at room temperature for 3 hours. The reaction mixture was poured into ethyl acetate (100 mi) with vigorous stirring. The resulting precipitate was collected by filtration, washed with a small 50 volume of ethyl acetate and dried to give 2.06 9 (100%) of the title compound X11-1K as a tan powder. Mp. 170-180'C (dec.).

IR: v,,,,, (KBr) in cm-1 1780, 1725, 1675, 1615, 1530, 1385, 1225, 1040, 750, 700.

UV: A ax (C2H,OH) in nm (E'%) 290 (196).

1 CM Column, Lichrosorb W18, 4x300 mm: Mobile phase, 0.01 M phosphate buffer (pH 7.2)/CH3OH=85/15: 60 Detection, uv (254 nm).

41 GB2194790A 41 NIVIR: J (DIVISO+D,0) in ppm 3.7 (2H, br.s, 2-H), 3.91 (3H, s, OCH3), 4. 70 (2H, s, Py-CH2-01-1), 5.28 (2H, m, CH,-N+), 5.23 (1H, d, J=5Hz, 6-H), 5.90 (1H, cl, J=51-1z, 7-H), 6.34 (1H, m, 3-CH=CM, 6.86 (1H, cl, J=1161- 1z, 3-CH), 6.89 (1H, s, CHPh,), 7.35 (10H, m, Ph-H), 7.9-8.9 (4H, m, Py- H).

B. 7-[2-(5-Amino-1,2.4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3hydrox ymethylpyridi- 10 nio)-1-propenyll-3-cephem-4-carboxylate (1-1K) (E isomer) A mixture of X11-1K (E isomer, 2.0 g, 2.4 mmoles) and sodium bisulfite (1 g) in 85% HCOOH (10 ml) was stirred for 2 hours at room temperature. The reaction mixture was concentrated to ca. 5 mi under reduced pressure. The oily residue was poured into acetone (100 mi) with vigorous stirring. The precipitate was collected by filtration, washed with a small amount of acetone and dried to give 1.1 g of a tan powder, which was purified by column chromatography (using the packing of a PrepPAK-500/C1, cartridge (Waters)] to give 283 mg (22%) of 1-1K as an amorphous powder. The powder was crystallized from 4N H.S04 and acetone to give 144 mg of the title compound I-1K as colorless needles. Mp. 185-188'C (dec.).

IR: vm,,,, (KBr) in cm-1 1775, 1680sh, 1660, 1630, 1225, 1045, 850.

UV: A (Phosphate buffer, pH 7) in rim (E'%) 236.5 (283), 275 sh (280), max 'em) 292.5 (330).

NIVIR: (5 (D20) in ppm 3.75 (2H, s, 2-H), 4.18 (3H, s, OCH3), 4.97 (2H, s, Py-CH2OH), 5.35 (1 H, d, Jz, 6-H), 5.43 (2H, d, J=6.51-1z, CH2-NI), 5.92 (1E, cl, J=41-1z, 7-H), 6.18 (1H d-t, J=16Hz, J=6.5Hz, 3-CH=CH-), 6.9 (1H, cl, J=16Hz, 3-CH), 8.13 (1H, d-d, J=8Hz, J=61-1z, Py-H), 8.60 (1H, cl, J=81-1z, Py-H), 8.84 (1H, d, J=61-1z, Py-H), 8.90 (1 H, s, Py-H).

Example 21

N-F C-CONN S 0 G H2Ni,'S'-, H K, 0 N CH=CH-CH,)-N3CONHC OCH3;;: e 2 coo I-1m E 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-(Z)-methoxyiminoacetamidol-3-[3(4-N-methylcarbamoy lpyri- 45 dinio)-1-propenyll-3-cephem-4-carboxylate (1-1M) (E isomer) A mixture of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2- methoxyiminoacetamido1-3(3-iodo-l-propenyi)-3-cephem-4-carboxylate (IX-1) (E isomer, 450 mg, 0.62 mmole) and 4-N methylcarbamoylpyridine [prepared according to the procedure of M. Samejima, Yakugaku Zasshi, 80, 1706 (1960)l (215 mg, 1.58 mmoles) in acetonitrile (2 m]) was stirred under nitrogen atmosphere for 5 hours at room temperature. The mixture was evaporated under reduced pressure and the residue was triturated with ether to give 530 mg of the quaternary salt. A mixture of the solid and sodium bisulfite (150 mg) in 85% formic acid (2 m]) was stirred for 4 hours and then heated at 40'C for 30 minutes. The mixture was evaporated under reduced pressure. The residue was triturated with acetone and the crude product was collected by 55 filtration. The crude product was chromatographed on a column of HP-20 (1. 5 x 18 cm) and the column was eluted with water and 30% aqueous methanol. The methanolic eluate was evapo rated under reduced pressure and the residue was freeze-dried to give 140 mg of an amorphous powder, which was further purified by HPLC (Column: Lichrosorb RP-18, Solvent: 15% CH3 OH) and the eluate of HPLC was freeze-dried to give 60 mg (18%) of the title product 1-1M. Mp. 180-183'C (dec.). Estimated purity: 80%.

IR: vr,,;,,, (KBr) in cm-1 1760, 1660, 1600.

42 GB2194790A 42 UV: A,,,,x (Phosphate buffer, pH 7) in nm (e) 230 (22100), 286 (22100).

NIVIR: 3 (D20) in ppm 3.08 (3H, s, CONHCH,), 3.72 (2H, s, 2-H), 4.16 (3H, s, OCH3), 5.35 (1H, d, J=4.5 Hz, 6-H), 5.95 (1H, d, J=4.51-1z, 7-H), 7.00 (1H, cl, J16Hz, 3-CH), 8.35 (2H, cl, J-6Hz, pyridine-H), 9.05 (2H, d, J=161-1z, pyridine-H).

Example 22

H 11 -CONN H 5 G177\ ) SO, H ' 0 0 - CH=CH-1m COOH M2H CH3 n H nne W_ i-1N ElZ=7/1 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(4carboxypyridinio)- 1-prope20 nyll-3-cephem-4-carboxylate (I- M) To a stirred suspension of isonicotinic acid (340 mg, 2.8 mmoles) in dry DMF (3.5 ml) was added N,O-bis(trimethylsilyi)acetamide (0.7 ml, 2.8 mmoles) under nitrogen atmosphere. To the resulting clear solution was added diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido]-3-(3-iodo-l-propenyl)-3-cephem-4-carboxylate (IX1) (E isomer, 720 mg, 1 mmole) in one portion, and the red solution was stirred for 1.5 hours at room temperature. The 25 reaction mixture was added dropwise to a stirred saturated sodium chloride solution (50 ml) containing sodium thiosulfate (150 mg). The yellow precipitate was collected by filtration, washed with water, and dried to obtain 772 mg of a pale yellow powder. The powder (700 mg) and sodium bisulfite (70 mg) were dissolved in 85% formic acid (5 ml), and the solution was allowed to stand at room temperature for 1.5 hours. The mixture was suspended in toluene (50 30 ml) and concentrated. The residue was triturated with acetone (70 ml), and the precipitate was isolated by filtration to afford 421 mg of a yellow powder. This crude powder (400 mg) was suspended in water (2 ml) and to the suspension was added sodium bicarbonate, The resulting dark solution was adsorbed on a column of the packing (50 ml) of a PrepPAK/C,, cartridge (Water's System 500), and the column was eluted by water (200 ml). The eluent was fraction- 35 ated into 10 fractions (20 ml of each), and the desired fractions (Fractions Nos. 4-7) were combined, acidified to pH 3 with 2N hydrochloric acid, and concentrated. The residue was triturated with acetone (30 ml) and the precipitate was collected by filtration to give 201 mg (37%) of the title compound I-1N as a yellow powder. E/Z=7/1; 80% pure. Mp. >189'C (dec.).

IR: vm,. (KBr) in cm - 1 1770, 1665, 1600.

UV: Amax (Phosphate buffer, pH 7) in nm (e) 227 (22500), 290 (22100).

45 NIVIR: (5 (D20+NaHCO3) in ppm 3,7 (2H, br.s), 4.15 (3H, s), 5.32 (1H, d, J=4Hz), 5.39 (2H, cl, J=61-1z), 6.14 (1H, d-t, J=15.5 and 6Hz), 7.03 (1 H, cl, J=15.51-1z), 8.31 (2H, cl, J=71-1z), 50 8.94 (2H, cl, J=7Hz).

Example 23

S 55 H --- CONH IN' N 11 1 CH=CH-CH np n_ M2H S ON3 0 coo E) 2 60 1-10 E 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-(Z)-methoxyiminoacetamidol-3-[3(2,3-cyclopentenopy ridi65 nio)-1-propenyll-3-cephem-4-carboxylate (1-10) (E isomer) 43 GB2194790A 43 A mixture of diphenyimethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetamido1-3(3-iodo-l-propenyl)-3-cephem-4-carboxylate (IX-1) (E isomer, 450 mg, 0,62 mmole) and 2,3cyclopentenopyridine (217 mg, 1.83 mmole) in acetonitrile (2 mi) was stirred under nitrogen atmosphere for 4 hours at room temperature. After evaporation under reduced pressure, the mixture was triturated with ether to give 560 mg of the quaternary salt. A mixture of the solid and 85% formic acid (2 m[) was stirred under nitrogen for 3 hours at room temperature and then heated at 40'C for 30 minutes. The mixture was evaporated under reduced pressure and trituration of the residue afforded 391 mg of the crude product, which was purified by chromatography on a column of HP-20 (1.5 x 18 em). The column was eluted with water and 30% aqueous methanol. Evaporation of the methanolic eluate under reduced pressure, followed by 10 freeze-drying afforded 160 mg of an amorphous powder, which was further purified by HPLC (Column: Lichrosorb, Solvent: 10% CH,OH). The eluate of HPLC was freeze-dried to give 50 mg (15%) of the title product 1-10. Mp. > l9O'C (dec.). Estimated purity: 75%.

* IR: vmax (KBr) in em-' 1765, 1670, 1600.

UV: A (Phosphate buffer, pH 7) in nm (e) 235 (20000), 283 (25000).

NMR (1),O+NaHCO.) in ppm 2.2-2.6 (2H, m, -CH2_), 3.1-3.6 (4H, m, -CH2_), 3.72 (2H, s, 2-H), 4.17 (3H, OCH3), 5.33 (1H, d, J=4.5Hz, 6-H), 5.90 (1H, d, J=4.5Hz, 7-H), 6.75 (1H, d, J=16Hz, 3-CH), 7.65-8.2 (3H, m, pyridine-H).

Example 24

H 5 N (D 5,N N \0 0 CH=CH-CH N, N2NJ 2 COOH e 1-2N C2H5 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-(Z)-ethoxyiminoacetamidol-3-[3-(4carboxypyridinio) - 1-prope nyll-3-cephem-4-carboxylate (1-2N, E isomer) and 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(Z)-ethoxyiminoacetamidol-3-[3-(4car boxypyridinio)-1-prope- 40 nyll-3-cephem-4-carboxylate (1-2N, Z isomer) To a chilled mixture of BSA (1.0 mi, 4.12 mmoles) and isonicotinic acid (506 mg, 4.12 mmoles) was added IX-2 (from Preparation No. 21) (1.0 g, 1.37 mmoles), and the mixture was stirred under nitrogen at room temperature for 2 hours. The mixture was poured into 10% Na2S203 (20 mi) of precipitate 1.3 g of the quaternary salt, which was collected by filtration, washed with water and dried. A mixture of the solid and sodium bisulfite (0.3 9) in formic acid (98%, 5 m]) was heated at 40'C for 1 hour and evaporated under reduced pressure. The residue was triturated with acetone and filtered to give 900 mg of the crude produce (E-propenyl isomer: Z-propenyl isomer=2:1). Separation of the isomers was carried out by HPLC (Column:

Lichrosorb, Solvent: 15% CH30H). The faster moving fractions of HPLC were collected, evapo- 50 rated under reduced pressure and freeze-dried to give the E-propenyl isomer of 1-2N (44 rng, yield 6%). The slower moving fractions gave the Z-propenyl isomer of 1-2N (32 mg, yield 4%) in a similar procedure.

1-2N, E isomer Mp.: >200'C (dec.). IR: vm;,,, (KBr) in em-' 1765, 1660, 1620, 1380.

UV: -A (Water) in nm (e) 228 (22200), 291 (23600).

44 GB 2 194 790A 44 NMR: 8 (D20) in ppm 1.45 (3H, t, J=6Hz, CH,CHJ, 3.72 (2H, s, 2-H), 4.45 (2H, q, CH2CH3), 5.40 (1H, cl, J=4Hz, 6-H), 5.90 (1H, cl, J=4Hz, 7-H), 7. 05 (1H, cl, J=15Hz, 3-CH), 8.30 (2H, cl, J=6Hz, Py-H), 8.95 (2H, cl, J=61- 1z, Py-H).

1-2N, Z isomer Mp.: >200'C (dec.).

IR: v,,, (KBr) in cm-1 1760, 1660(sh), 1620, 1370.

UV: Az (Phosphate buffer, pH 7) in rim (e) 225 (22400), 275 (sh, 16000).

NMR: (5 (D20) in pprn 1.45 (3H, t, J=7Hz, CH2CH3), 3.50 (11-1, cl, J17Hz, 2-H), 3.75 (11-1, cl, J=171-1z, 2-H), 5.38 (1H, cl, J=4Hz, 6-H), 5. 95 (1H, cl, J = 4Hz, 7-H), 6.62 (1 H, d, J = 11 Hz, 3-CH), 8.35 (2H, cl, J=6Hz, Py-H), 8.92 (2H, cl, J=6Hz, Py-H).

Example 25 S

N C-CONN N H N f CONN2 N2N s', \D 0 CH=CH-CHN G 2 ==r 1-3H H2- CH: C H 2 E 7-[2-Amino-1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)-acetamidol-3-[3(4 -carbamoylpyridinio)-1- 30 propenyll-3-cephem-4-carboxylate (1-3H) (E isomer) To a solution of 35 mg (0.08 mole) of 7-amino-3-[3-(4-carbamoylpyridinio)- 1-(E)-propenyi]-3cephem-4-carboxylate hydrochloride in 2 mi of 50% aqueous acetone was added 52 mg of 2-[5 amino1,2,4-thiadiazol-3-yi)]-2-(propen-3-yioxyimino)acetyl chloride hydrochloride (from Preparation No. 25) and the mixture was adhusted to pH 6.5-7.0 with 2N Na2C03. The mixture was stirred at room temperature for 1 hour, acidified to pH 2 with 1N HCl and concentrated under reduced pressure. The residue was chromatographed on a column of HP-20 resin which was eluted with 300 mi of water and 30% CH301-1-1-120. Fractions containing the product were combined and evaporated under reduced pressure. The residue, 73 mg, was purified by a column of reverse phase carrier which was taken out of a PrepPAK-500/C, cartridge (Waters, 30 mi). The column 40 was eluted with water, 5% CH30H, 10% CH30H and 20% CH30H, successively. Fractions containing the product were combined and lyophilized to afford 26 mg (62%) of the title product 1-31-1. Mp. 160'C (dec.).

IR: Vmax (KBr) in pm- 1 3400, 1765, 1680, 1605, 1400. 45 UV: A,,,, (Phosphate buffer, pH (7) in nm (e) 226 (24600), 288 (22800).

NMR: (5 (D20) in ppm 3.75 (2H, s, 2-H), 5.41 (1H, cl, J=51-1z, 6-1-1), 5.50 (41-1, m, C1-12NI, & CH=CH2), 5.98 (1H, cl, J=51-1z, 7-H), 6.20 (11-1, m, 3-(CH=CM, 7.09 (1H, d, J=17Hz, 3-CH), 8.50 (2H, cl, J=7Hz, Py-H), 9.16 (21-1, cl, J=7Hz, Py-H).

Example 26

N----F C-CONN 60) A N N N2N 5 1\0 0 CH--CH-CH2- -CONH2 1 -CH C000 Rl=r CH2C 1-41-1 E GB2194790A 45 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetamidol-3-[3(4-carbamoylpyridi nio)- 1propenyll-3-cephem-4-carboxylate (1-4H) (E isomer) To a solution of 86 mg (0.19 mmole) of 7-amino-3-[3-(4carbamoylpyridinio)-1-(E)-propenyll-3cephem-4-carboxylate hydrochloride (XXII-H) in 2 mI of 50% aqueous acetone was added 63 mg of 2-propargyloxyimino-2-(5-amino-1,2,4-thiadiazol-3-yi)-acetyI chloride hydrochloride (from Preparation No. 26). The suspension was maintained at pH 6.5-7.0 with 2N Na2C03 and then stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 2 with 1N HCl and concentrated in vacuo. The residue was diluted with 30 m[ of water, neutralized with NaHCO, and filtered. The filtrate was transferred on the top of a column which was packed with reverse phase carrier (30 mi) taken from a PrepPAK-500/C, cartridge (Waters). The column was eluted with water, 5% CH,OH, 10% CH30H and 20% CH30H, successively. Fractions containing the product were combined and lyophilized to afford 13 mg (12%) of the title product 1-4H. Estimated purity 70%. Mp. 1600C.

IR: v,,a,, (KBr) in cm-1 3400, 2120, 1765, 1680, 1610.

UV: An,, (Phosphate buffer, pH 7) in nm (e) 229 (24000), 288 (21200).

NMR: (5 (D,O) in ppm 3.78 (2H, s, 2-H), 5.15 (2H, d, J = 1 Hz, -CH2-C-CH), 5.40 (1H, d, J=5Hz, 6-H), 5.50 (2H, m, CH-N+), 5.98 (1H, cl, J=5Hz, 7-H), 6.20 (1H, m, 3-CH=CM, 7.05 (1H, cl, J=171-1z, 3-CM, 8.50 (2H, d, J=71-1z, Py-H), 9.16 (2H, d, J=7Hz, Py-H).

Example 27 5

N OF H CONN2 H2H S''N 0 --CH--CH-H2-H 1-5H rIIII 'E M Ole 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetamidol-3[3-(4-carbamoylpyri dinio)- 1 pro,dignyll-3-cephem-4-carboxylate (1-5H) (E isomer) To a stirred solution of 139 mg (0.31 mmole) of 7-amino-3-[3-(4carbamoylpyridinio)-1-prope- nyll-3-cephem-4-carboxylate hydrochloride in 3.5 mi of 50% aqueous acetone in an ice-cooling bath was added portionwise 120 mg (0.44 mmole) of 2-(5-amino-1,2,4- thiadiazol-3-yl)-2-cyclopentyloxyiminoacetyl chloride hydrochloride (from Preparation No. 27). The mixture was adjusted to pH 6.5-7.0 with 2N Na2C03 (0.9 mi) and stirred for 1 hour at 10'C. The reaction mixture was acidified to pH 2 with 1N HCl and evaporated under reduced pressure. The residue was chromatographed on a column of HP-20 resin (20 mi) and was eluted with 300 mi of water and 30% CH,OH-H20, successively. Fractions containing the product were combind and concentrated 45 in vacuo. The residue was treated with 60 m] of acetone to give 111 mg (83%) of the title compound 1-5H. Mp. 160'C (dec.). Estimated purity 70%.

IR : Vmax (KBr) cm 1 3400, 1770, 1680, 1605, 1530.

UV: A (Phosphate buffer, pH 7) in nm (e) 224 (23300), 286 (24600).

46 GB2194790A 46 NMR: (5 (DIMSO-Q in ppm 1.70 (81H, br.s, H]), 4.68 (1 H, br.s, 5.05 (11H, cl, J=5Hz, (6-H), 5.30 (2H, m, CH,W), 5.67 (1H, 10 cl-cl, J=51Hz & 7Hz, 7-H), 6.20 (11H, m, 3-CH=CM, 7.08 (11H, cl, J=171Hiz, 3-CH), 8.34 (2H, cl, J=7Hz, Py-H), 9.11 (2H, d, J=7Hz, Py-H), 9.38 (1H, cl, J=7Hz, 7-NH). 15 Example 28

N C-CONN 5 --IF 11 Nk SN N, 0 N --- CH=CH-CH2-Nd OCH3 T COOE) 1-1 P 0 H2COOH E 7-[2-(5-Amino1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol-3-[3-(3carboxymethylpyridin io)- 1propenyll-3-cephem-4-carboxylate (1-1P) (E isomer) A. DiphenyImethyl 7[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-[3-(3-carbox 30 ymethylpyridinio)- 1-propenyll-3cephem-4-carboxylate (X11- 1P, iodide, E isomer) To a suspension of 3-carboxymethylpyridine hydrochloride (0.89 9, 5 mmoles) in 10 mi of CH,Cl, was added N,O-bis(trimethyisiiyi)acetamide (4. 97 mi, 18 mmoles), and the mixture was stirred at room temperature until a clear solution was obtained. To the solution was added IX-1 (1.79 g, 2. 5 mmoles), and the mixture was allowed to stand at room temperature. After 3 hours, 3 mi of CH30H was added to the cooled mixture and the solution was evaporated in vacuo to give an oil which was triturated with ethyl acetate to afford 2.28 9 of the title compound X11-11P as a tan powder. Mp. 161'C (dec.).

IR Vmax (KBr) in em 1780, 1720, 1675, 1630, 1530, 1385, 1 225, 1045, 755, 700.

UV: A (C,H,OH) in nm (E'%) 295 (188).

max 1 em NIVIR: J (DIMSO+D20) in ppm 3.70 (21-1, br.s, 2-H), 3.90 (5H, s, OCH, & Py-CH,CO), 5.25 (3H, m, -CH2N 1 & 6-H, 5.92 (1 H, cl, J=4.51Hiz, 7-H), 6.35 (1H, rn, 3-CH=CH-), 6.90 (1H, cl, J=16Hz, 3-CH), 6.92 (1H, s, CHPh2), 7.35 50 (10H, m, Ph-H), 8.8-9.0 (4H, m, Py-H).

B.7-[2-(5-Amino-.1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(3car boxymethylpyridinio)- 1-propenyll-3-cephem-4-carboxylate (1- 1(E isomer) A mixture of X11-11P (iodide) (2.28 9) and sodium bisulfite (1.1 9) in 85 HCOOH (10 mi) was stirred at room temperature for 2 hours. The reaction mixture was concentrated to ca. 5 mi under reduced pressure. The oily residue was triturated with acetone (100 m]) to give 1.22 9 of the crude product, which was purified by column chromatography (HP-20, 420 mi) to afford 533 mg of the title compound 1-11P (38%, from W-11) as a pale yellow amorphous powder. Mp. 60 1WC (dec.).

IR: vm,,x (KBr) in em-' 1770, 1670, 1600, 1530, 1385, 1140, 1040.

47 GB2194790A 47 UV: 2. (Phosphate buffer, pH 6.2) in nm (E'%) 234 (374), 277sh (390), 290 (402).

max 1 CM NMR: (5 (D,O+NaHCO.) in ppm 3.78 (2H, s, 2-1-1), 3.92 (2H, s, Py-CH,CO), 4.22 (3H, s, OCH3), 5.40 (1H, cl, J=41-1z, 6-H), 5.44 (2H, d, J=6.51-1z, -CH.-N-1), 5.97 (11-1, d, J=41-1z, 7-H), 6.20 (1H, d-t, J=16 & 6.5Hz, 3-CH=CM, 10 7.08 (11-1, d, J=161-1z, 3-CH), 8.11 (11-1, d-d, J=8 & 7Hz, Py-H5), 8.53 (11-1, d, J=81-1z, Py-HJ, 8.82 (1H, d, J=71-1z, Py-H.), 8.86 (11-1, s, Py-1-12).

Example 29

S 11 11 N N (D g,[-- N ' N OCH3 coo E) 1-1Q m C-CONH E 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-[3(4carboxymethylthiopyr idinio)- 1-propenyll-3-cephem-4-carboxylate (1-1Q) (E isomer) A. Diphenyimethyl 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-[3-(4-carbox- ymethytthiopyridinio)- 1 -propenyll-3-cephem-4-carboxylate (Xll- 1 Q, iodide, E isomer) To a suspension of 4-carboxymethylthiopyridine (0.88 g, 5 mmoles) in 10 mi of CH,Cl, was added N,O-bis(trimethyisiiyi)acetamide (5 mi, 18 mmoles), and the mixture was stirred at room temperature until a clear solution was obtained. To the solution was added IX-1 (E isomer 1.79 g, 2.5 mmoles) and the mixture was allowed to stand at room temperature. After 3 hours, 3 m] of CH30H was added to the cold mixture and the solution was evaporated in vacuo to give oily residue which was triturated with ethyl acetate to give 2.43 g of the title compound X11-1Q (iodide) as a tan powder. Mp 155T (dec.).

IR: v,,,, (KBr) in cm--' 1780, 1720, 1670, 1625, 1525, 1385, 1225, 1115, 1040, 755, 700.

uv: Amax (C2H5OH) in nm (E'%) 312 (299).

1 CM NIVIR: 6 (DMSO-d6+D20) in ppm 3.70 (2H, br.s, 2-H), 3.93 (31-1, s, OCH3), 5.07 (21-1, m, CH2-NI.), 5.23 (11-1, d, J=SHz, 6-1-1), 5.90 (11-1, d, J=51-1z, 7-H), 6.29 (11-1, m, 3-CH=CM, 6.87 (11-1, d, J=161-1z, 3-CH), 6.91 (11-1, s, CHPh2), 7.35 (10H, m, Ph-H), 7.88 & 8.58 (each 2H, cl, J=6Hz, Py-H).

B. 7-[2-(5-Amino- 1,2,4-thiadiazol-3yl)-2-methoxyiminoacetamidol-3-[3-(4carboxymethylthiopyr i55 dinio)- 1-propenyll-3-cephem-4-carboxylate (I1Q) (E isomer) A mixture of X11-1Q (iodide, 2.43 9) and sodium bisulfite (1.1 9) in 85% HCOOH (10 m]) was stirred at room temperature for 2 hours. The reaction mixture was concentrated to ca. 5 mi under reduced pressure. The oily residue was triturated with acetone (100 mi), filtered and dried to give a crude product (1.39 g), which was purified by column chromatography (HP-20, 20 mi) to afford 577 mg of the title compound 1-1Q (39% from W-1) as a pale yellow amorphous 60 powder. Mp. 18WC (dec.).

[R: v.,,, (KBr) in cm-1 1765, 1670, 1625, 1530, 1380, 1110,1035.

48 GB2194790A 48 UV: A (Phosphate buffer, pH 6.2) in nm (E'%) 234 (459), 310 (678).

max 1 cm NMR: 8 (13,0+NaHCO,) in ppm 3.79 (2H, br.s, 2-H), 4.10 (2H, s, S-CH2), 4.23 (3H, s, OCH3), 5.25 (2H, d, J=6.5Hz, CH27W), 5.39 (1H, d, J=4.0Hz, 6-H), 5.97 (1H, cl, J=4Hz, 7-H), 6.18 (1H, d-t, J=15.5Hz & 6.5Hz, 3-CH=CM, 10 7.05 (111, d, J=15.51-1z, 3-CH), 7.84 & 8.55 (each 2H, d, J=7Hz, Py-H).

Example 30

H C CONH - S 1 r- H3C G H2Nk.'M N 04: N CH--CH-CH2-N \OCH3 0 G coo E/Z=7/1 1-1A 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(lmethylpyrrolidinio)- I-propenyll-3-cephem-4-carboxylate sulfate (/- 1A, sulfate) A. Dipheny/methyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-[3-(I-methyl pyrrolidinio)-1-propenyl]-3-cephem4-carboxylate (XII-1A, iodide) To a cold solution of diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)2-methoxyiminoacetamidoj-3-(3-iodopropenyl)-3-cephem-4-carboxylate (IX-1) (from Preparation No. 14) (21.5 g, 30 mmoles) in ethyl acetate (300 ml) was added dropwise a solution of 1-methylpyrrolidine (2.55 g, 30 mmoles) in ethyl acetate (30 ml) over a period of 1 hour at -5 to O'C, with stirring. After stirring for an additional 10 minutes, the resulting precipitate was collected by filtration and washed with chloroform (200 ml) to give 23.0 g (95.8%) of the title compound (IX-1A, iodide), melting at >175'C (dec.).

IR: vmax (KBr) in em 1 3300, 1780, 1730, 1685, 1615.

UV:), (CH,OH) in nm (E'%) 218 (435), 295 (188).

max 1 cm B. Diphenyimethyl 7-[2-(5-Amino- 1, 2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-[3-(1 -methylpyrrolidinio)-1-propenyll-3-cephem4-carboxylate (X11-1A, chloride) The compound (X11-1A, iodide) (23 9, 28.7 mmoles) was dissolved in a mixture of acetone and methanol (1A, 230 mi) and applied on an Amberlite IRA-410 (chloride form, 230 mi) column which was pretreated with the same mixed solvent. The column was developed with the 45 solvent and the fractions containing the desired compound were combined and concentrated to an oily residue, which was triturated with ethyl acetate (300 mi) to yield 17.9 9 (87.7%) of the title compound (X11-1A, chloride), melting at 1900C (dec.).

IR: (KBr) in cm 1 3380, 1780, 1680, 1620.

UV: A (C,H,OH) in nm (E'%) 220 (369), 290 (232).

max 1 cm C. 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3-(1methyl pyrrolidinio)-1- 55 propenyll-3-cephem-4-carboxylate sulfate (1-1A, sulfate) A mixture of the compound (X11-1A, chloride) (17.8 9, 25 mmoles) in 85% formic acid (178 mi) was stirred at room temperature for 2 hours under a nitrogen atmosphere. The mixture was evaporated in vacuo and the oily residue was triturated with acetone to give 9.80 9 of crude 1-1A. Concentration of the filtrate and the acetone washings yielded additional 2.95 g of crude 60 1-1 A. Two crops of the crude material were combined and extracted with 2N HQ (1 L and 0.5 L). The combined extracts were adsorbed on a Diaion HP-20 resin (1.5 L column), which was eluted with water and 30% aqueous methanol. The desired fractions were collected and evapo rated in vacuo to an oily residue, which was triturated with isopropanol (200 mi) and acetone (200 mO, successively, to yield 7.09 9 of a light yellow powder. This material (6.80 g) was 49 GB2194790A 49 dissolved in water (20 ml) and then subjected to column chromatography over the packing of PrepPAK-500/C,, cartridge (90 ml), using water and 10% aqueous methanol as an eluent. The eluate was collected in 20-ml fractions with monitoring by HPLC. [Column, Nucleocil SSC-ODS-262, 8 x 100 mm; Mobile phase, 0.0 1 M phosphate buffer (pH 7.2)/CH,OH =90: 10; Detection, UV (254 nm)]. Fraction No. 4 through Fraction No. 10 were combined, evaporated under reduced pressure and Iyophilized to give 2.28 g of a yellow powder (E/Z=7/1, 70% pure) [Crop 1]. Fraction No. 11 through Fraction No. 85 were worked up in the same manner -as described above to give 3.27 g of yellow powder (E/Z=5/1, 70% pure) [Crop 2]. A portion of Crop 1 (1.0 g) was purified by rechromatography on the packing of PrepPAK-500/C,, cartridge (90 ml). The column was eluted with water and 5% aqueous methanol, successively. The eluate 10 containing the desired compound was concentrated and Iyophilized to yield 638 mg (E/Z7/1, 80% pure) of yellow powder. Another portion of Crop 1 (1. 14 g) was worked up the same way to give 880 mg (E/Z=7/1, 80% pure) of yellow powder. The two purified samples were combined and a portion (1.45 g) dissolved in 1N sulfuric acid (5 ml). The solution was diluted with acetone (315 ml), with stirring. The creamy precipitate was collected by filtration to obtain 15 1.48 g of the title compound (I-1A, sulfate) (E/Z=7/1, 80% pure), melting at >185'C (dec.).

IR Vinax (KBr) in cm-1 3380, 3000, 1765, 1675, 1630, 1535, 1390, 1115.

UV: (Phosphate buffer, pH 7) in nm (e) 236 (19900), 291.5 (22500).

NMR: (5 (D20+NaHCO3) in ppm 2.36 (4H, br., N,'), 3.15 (3H, s, CH, -ICH3 NJ), 3.62 (5 H, 30 br., 2-H and I), 3.83 (1 H, br., 2-H), 4.13 (2H, d, J=8Hz, CH2N,), 4.22 (31-1, s, OCH3), 5,39 35 (1H, d, J=4.5Hz, 6-H), 5.96 (11-1, d, J=4.5Hz, 7-H), 6.00 (11-1, m, 3-CH=CH, 6.67 (1/8H, d, J=lOHz, 3-CH, cis), 7.04 (7/8H, d, J=16Hz, 3-CH, trans). 40 Example 31

S H C-CONH HNjS.' 4 0 9 N (CH3)3 2 N\OCH3-7i-"?e CHI'CH-CH2- coo 1-1 D E/Z=10/1 50 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-[3- trimethylammonio1propenyll 3-cephem-4-carboxylate (/- W) A. Diphenyimethyl 7-[2-(5-amino1,2,4-thiadiazol-3-yl)-2- methoxyiminoacetamidol-3-(3-trimethylammonio-l-propenyl)-3-cephem-4-carboxylate (X11-1D, iodide) To a solution of 13.0 9 (19 mmoles) of diphenyimethyl 7-[2-(5-amino-1,2,4thiadiazol-3-yi)-2methoxyiminoacetamido]-3-(3-iodopropenyl)-3-cephem-4-carboxylate (IX-1, from Preparation No.

10) in 38 m] of dry ethyl acetate was added 1.75 mi (19.1 mmoles) of 1. 1 N trimethylamine in ethyl acetate at -WC, and the mixture was stirred for 1 hour at -5'C. The resulting precipitate was filtered off, washed well with CHCI, and dried to give 12.5 g (88%) of the title compound 60 (X11-1D) as the iodide.

IR: vm,, (KBr) in em-' 3300, 1765, 1720, 1665.

GB2194790A 50 UV: (C^OH) in nm (e) 300 (18400).

B. Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-(3-trimethylammonio-l-propenyl)-3-cephem-4-carboxylate (X11-1D, chloride) The iodide (X11-11D, 12,5 9) was dissolved in 60 m[ of CH,OH-acetone (M) and passed 5through a column of ion-exchange resin [MA-410 (C]-), 125 m]]. The column was eluted with 300 mi of CH,OH-acetone (1:1), and the eluate was evaporated in vacuo and triturated with 300 mi of isopropyl-ether to afford 10.4 9 (91%) of the quaternary salt (X11- 11D, chloride).

IR: vr (KBr) in cm-1 3300, 1765, 1710, 1665.

UV: Amax (C2H5OH) in nm (e) 298 (15100), C. 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidoj-3-[3- trimethylammonio- 1 -pro penyll-3-cephem-4-carboxylate (/- 1D, sulfate, E isomer) A solution of 10.4 g (16.0 mmoles) of Xll-1D (chloride) in 20.8 ml of 85% formic acid was allowed to stand for 3 hours at room temperature and concentrated in vacuo. The residue was treated with 210 ml of isopropanol and the precipitate was filtered off. The solid (10.1 g) was triturated with 210 ml of water and neutralized with sodium bicarbonate. The suspension was filtered off and the filtrate was chromatographed on a column of HP-20 (300 ml) which was 20 eluted with water (1000 ml), 10% CH30H (200 ml) and 30% CH30H (150 ml), successively.

Fractions containing the desired product were combined and concentrated under reduced pres sure. The residue was purified by reverse phase chromatography. The column was packed with a packing obtained from a PrepPAK-500 C1, cartridge (Waters, 200 ml). Elution with water (600 ml) and 30% CH30H (200 ml), successively, followed by concentration of fractions containing the 25 desired product gave 2.52 g (18%) of the title compound. A solution of the zwitterionic product (1.5 g) in 1N H2SO4 (5 ml) was added portionwise to 300 ml of acetone and the resulting precipitate was filtered and dried. Yield of I-11D sulfate was 1.42 g (80%). The ratio of E/Z was approximately 10/1 based on HPLC.

IR: v,,,, (KBr) in cm- 1 3380, 1765, 1665.

UV: A (Phosphate buffer, pH 7) in nm (e) 237 (19500), 293 (22400).

4 NMR: 3 (D,O) in ppm 3.25 (9H, s, NI -CH,), 3.94 (2H, s, 35 2-H), 4.14 (2H, d, J=71-1z, CH2NI), 4.23 (3H, s, O-CHJ, 5.42 (1H, d, J=4.5Hz, 6-H), 6.00 (1H, d, J=4.51-1z, 7-H), 6.23 (1H, d-t, J=7 & 16Hz, 3-CH=CM, 7.23 (1 H, d, J = 16Hz, 3-CH). 40 Example 32

N - ONH - 5 - J J CH= H2H OC3 0 r_ 4!X==/ coo E) 1-1 H E 7-[2-(5-Amino1,2,4-thiadiazol3-yl)-2-methoxyiminoacetamidol-3-[3-(4carbamoylpyridinio)- 1-propenyll-3-cephem4-carboxylate (1-1H, E isomer) To a mixture of 7-amino-3-[3-(4-carbamoylpyridinio)-1-(E)-propenyll-3cephem-4-carboxylic acid hydrochloride (XXII-H, 397 mg, 1 mmole) and NaHCO, (168 mg, 2 mmoles) in aqueous DIVIF (water, 3.5 m] and DIVIF, 7 5 mi) was added benzotriazol-lyi-2-(5-amino-1,2,4-thiadiazol-3-yi)-2methoxyiminoacetate (479 mg, 1.5 mmoles) (from Preparation No. 28). The mixture was stirred at room temperature for 3.5 hours. The reaction mixture was adjusted to pH 3-4 with 3N HCl and diluted with 200 mi of acetone to give a precipitate, which was collected by filtration. The crude product was dissolved in a small volume of aqueous THF and the solution was adjusted to pH 6.8 with NaHCO, treated with decolorizing carbon, concentrated to ca. 1 mi and seeded with a few pieces of crystalline 1-1H. After stirring overnight, the crystalline precipitate was collected by filtration to afford the title compound 1-1H (zwitterion form). Yield 83 mg (16%). Mp. >185'C (dec.). Physico-chemical data of this product were identical to those of the compound in Example 10.

51 GB2194790A 51 Preparation No. 1 Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-chloromethyi -3cephem-4-carboxylate (IV- 1) To a stirred suspension of 2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetic acid (111-1) (2.1 9, 10 mmole) in dry CH2C12 (50 H) was added PC1, (2.09 9, 10 mmole) at -300C, and the mixture was stirred for 20 minutes at -15 to -20'C. To the above acid chloride solution was added a solution of diphenyimethyl 7-amino-3-chloromethyi-3-cephem-4- carboxylate hydrochloride (11) (4.5 9, 10 mmole) in CH2C12 (50 mi) containing N,O-bis-(trimethyisilyl)acetamide (10 g, 50 mmole) at -30'C. After stirring at -WC for 1 hour, the mixture was concentrated to remove 10 the CH2C12 and diluted with ethyl acetate (200 mi). The mixture was washed with 10% aqueous NaHCO, (2x40 mi), H20 (2x20 mi) and brine (10 mi), successively, and dried over M9S04. The solvent was evaporated in vacuo and the resulting oily residue (10 g) was dissolved in CHCI, (20 ml) and chromatographed on a silica gel (Wako gel C-200, 100 g containing 10 mI of 1/1.5 M pH 7 phosphate buffer) using 1-3% CH,OH-CHC13. Fractions containing the title compound was evaporated to give 5.7 g (95%) of IV-1 as a yellow amorphous powder. M. p. >l4WC (dec.).

M: VUr cm-1 3300, 1780, 1720, 1680, 1620.

max UV: ZEtOHnm (8) 245 (1800), 280 (9900). max NIVIR: 6MSO-d6 3.53 (2H, A13q, 2-H), 3.94 (3H, s, OCH,), 4.42 (2H, s, 3- CH2), 5.22 (1H, d, J=4.5, 6-H), 5.92 (1H, d-d, J=4.5 & 6, 7-H), 6.93 (1H, s, CHPh2), 7.36 (10H, rn, Ph-H), 8.1 (2H, br-s, NH2), 9.58 (1H, d, J=6, 7- NH).

Preparation No. 2 Diphenylmethyl 7-[2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-iodomethy]-3-ce- phern-4-carboxylate (V- 1) A mixture of IV-1 from Preparation No. 1 (5.7 9, 9.5 mmole) and Nal (4.3 9 29 mmole) in dry acetone (50 m]) was stirred for 5 minutes at room temperature. The mixture was concen- 35 trated under reduced pressure and the resulting oil was shaken with a mixture of ethyl acetate (100 mi) and H20 (lomi). The organic layer was separated and washed with 10% w/v) sodium thiosulfate and brine, successively. After drying, the ethyl acetate was removed in vacuo to give 6.1 g (93%) of the title compound (V-1) as a yellow amorphous powder melting at >l2WC (dec.). 40 M VUr cm-1 3300, 1780, 1725, 1680, 1620.

max UV: VEtOH nm (e) 245 (17000), 282 (12000).

max NIVIR: 6DMSO-d 6 3.72 (2H, ABq, 2-H), 3.94 (3H, s, OCH3), 4.23 PPM (2H, s, 3-CH2), 5.21 (1H, d, J=4.5, 6-H), 5.89 (1H, d-d, J=4.5 & 6, 7-H), 6.94 (1H, s, ChPh2), 1.35 (10H, m, Ph-H), 8.12 (2H, br-s, NH2), 9.65 (1H, d, J=6, 7-NH).

Preparation No. 3 Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-triphenylpho sphoniomethyl-3-cephem-4-carboxylate iodide (V1-1) A mixture of V-1 from Preparation No. 2 (6jO mg, 1 mmole) and triphenylphosphine (786 mg, 3 mmole) in ethyl acetate (20 mi) was stirred overnight at room temperature. The solid which separated was collected, washed with ethyl acetate (2 x 10 mi) and dried to give 950 mg (100%) of the phosphonium iodide VI-1. M. p. 18WC (dec.).

IR: VKBr cm-1 3300, 1760, 1710, 1680, 1610.

max 52 GB2194790A 52 UV: A110H nm (e) 268 (15000), 275 (13000), 300 (7300).

max NIVIR: &MSO-d6 3.52 (2H, br-s, 2H), 3.94 (3H, s, OCH,), 5.34 pprn (1H, d, J=4.5, 6-H), 5.9 (1H, m, 7H), 6.3 (1H, s), 7.3 (10H, m, Ph-H), 7.8 (15H, m, Ph-H).

Preparation No. 4 Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-[(triphenylp hosphoranylidene)methyll-3-cephem-4-carboxylate (VII- 1) A mixture of V1-1 from Preparation No. 3 (952 mg, 1 mmole), Amberlite IRA- 410 (OH- form, 500 mg) and N NaOH (4 mi) in CH,Cl, (10 mi) was stirred for 1 hour at room temperature. The mixture was filtered and the separated organic layer was dried over M9S04 and concentrated under diminished pressure. The resulting oil was triturated with ethyl acetate and the resulting yellow precipitate was collected by filtration to give 740 mg (90%) of the title compound V11-1. M.p. >l8WC (dec.).

IR VKBr cm-1 3400, 1750, 1630.

max UV: AEtOH nm (e) 268 (12000), 276 (10000), 384 (23000).

max Preparation No. 5 Diphenyimethyl 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2- methoxyiminoacetamidol-3-(3-chloro- 1-pro penl-yl)-3-cephem-4-carboxylate (V111-1) To a solution of V11-1 from Preparation No. 4 (6.9 9, 8.4 mmole) were added MgSO, (3 9) and 40% chloroacetaldehyde (810 mg, 8.4 mmole). The mixture was stirred for 1.5 hours at room temperature and then filtered. The filtrate was eluted on silica gel (Wakogel C-200, 100 9 30 containing 10 mi of 1/1.5 M phosphate buffer) column by using CHO, and CHC13 containing CH30H. Fractions containing the desired product (0.5-1% CH301-1CHC13) were evaporated in vacuo to give 1.6 9 (30%) of the title compound V111-1 as a yellow amorphous powder, which was a mixture of the Z and E isomers with respect to the chloropropenyl moiety (Z/E=2/1, by nrm). M.p. >l3WC (dec.). 1 35 IR VKBr cm-1 3300, 1780, 1725, 1680, 1620.

max UV: AE10H nm (e) 240 (20000), 286 (12000). 40 max NIVIR: 6DMSO-d6+D20 3.56 & 3.8 (m, 2-H), 3.94 (3H, s, OCH3), PPM 4.16 (d, J=7.5, CH20), 5.26 (1 H, cl, J=4.5, 6-H), 5.87 (1H, d, J=4.5, 7-H), 45 6.28 (213H), cl, J = 11, 3-CH cis-H), 6.72 (1 /3H, cl, J = 16, 3-CH trans-H), 6.81 (2/3H, s, ChPhA, 6.92 (l/3H, s, ChPh2), 7.4 (1 OH, m, Ph-H). - 50 Preparation No. 6 DiphenyImethyl 7-Benzylideneamino-3-[(triphenylphosphoranylidene)methyll3-cephem-4-carboxy late (XVI) To a solution of diphenyimethyl 7-benzyiideneamino-3- [(triphenylphosphonio)methyll-3-cephem4-carboxylate iodide (XV) [prepared according to the procedure of Japan published patent appli- 55 cation (Kokai) 56-86187 (7/31/81)l (60 9, 70 mmoie) in CH,Cl, (350 mi) were added N NaOH (140 mi) and Amberlite IRA-410 (OH form, 35 9) at 5'C. The mixture was stirred for 1 hour at WC and filtered. The organic layer was separated, dried over M9S04, concentrated to ca. 100 mi of volume and precipitated with ethyl acetate (500 mi). The resulting yellow solid was collected by filtration and dried in vacuo to afford 48 9 (94%) of the title compound XVI, 60 melting at 195-WC (dec.).

IR: VK13r cm-1 1770, 1620.

max 53 GB2194790A 53 Preparation No. 7 Diphenylmethyl 7-Benzylideneamino-3-(3-chloro-l-propen-l-yl)-3-cephem-4- carboxylate (XVII) To a stirred solution of XVI from Preparation No. 6 (2.9 g, 4 mmole) in a mixture of CH2C12 (40 mi) and H20 (10 mi), was added anhydrous chloroacetaldehyde (800 mg) at room tempera ture. To the mixture was added additional 800 mg of chloroacetaldehyde in three portions over 5 a period of 1 hour, while the pH of the mixture was kept between 6 to 9 by addition of N NaOH. After 15 minutes, the aqueous layer was removed and the organic layer was dried over MgSO, Evaporation of the solvent gave a red oil which was dissolved in a mixture of ethyl acetate and isopropyl ether (l/2, 80 mi). The solution was washed with saturated aqueous NaHCO, (10 mi) and H,0 (10 m]), successively. After drying over M9SO, removal of the solventO afforded 3.3 9 of yellow oil. A solution of the oil in CH2C12 (50 mi) was filtered with aid of silica gel (12 g, Wakogel C-200) containing 1/1.5 M phosphate buffer (1.2 mi, pH 6.4) and the silica gel was washed with CH2C12 (50 m]). The filtrate and washing were combined and evaporated to dryness. The residue was triturated with n-hexane to give 1.7 g (80%) of the title compound (XVII) as a yellow powder. The nmr spectrum indicated that the chloropropenyl moiety had the Z5 configuration. M.p. >5WC (dec.).

M: vKB, em-' 3400, 1775, 1720, 1630.

max UV: ZEtOH nm (e) 253 (11000), 258 (11000), 265 (10000), 273 max (8300), 281 (7000), 290 (6300).

NMR: 5DMSO-d. 3.63 (2H, br-s, 2-H), 4.0 (2H, m, CH2-CI), 5.42 ppm (2H, m, 6-H & 3-CH=CM, 5.72 (1H, d, J=4.5, 7-H), 6.27 (1 H, d, J = 11, 3-CH), 6.85 (1 H, s, ChPhA, 7.33 (10H, m, Ph-H).

Preparation of anhydrous chforoacetaidehyde Anhydrous calcium chloride was added to a chilled solution of 50% aqueous chloroacetal- dehyde (50 ml), with stirring, to separate it into two layers. The chloroacetaldehyde hydrate layer (1) (the upper layer) was separated and diluted with CHCI, (100 mi), mixed with MgSO, (20 g), heated to reflux for 5 minutes, and filtered. The solvent and water waere removed azeotropi cally (b.p. 56-64'C) (2), and the residue was distilled to give anhydrous chloroacetaldehyde (3), 35 b.p. 70-82'C/760 mm.

IR: 01- cm - 1 1720.

max (1) R.P. Kurkjy, E. V. Brown, J. Amer. Chem. Soc., 74, 5778 (1952). (2) S. Trippet, D. M. Walker, J. Chem. Soc., 1961 1266. (3) H. 0. House, V. K. Jones, G. A. Frank, J. Org. Chem., 29, 3327 (1964).

Preparation No. 8 Diphenylmethyl 7-Amino-3-(3-chloro-l-propen-1-yl)-3-cephem-4-carboxylate (XV111) A solution of XVII from Preparation No. 7 (180 mg, 0.34 mmole) in ethyl acetate (10 ml) was added to a solution of Girard Reagent T [(carboxymethyl)trimethylammonium chloride hydrazide] (251 mg, 1.5 mmole) in CH30H (10 ml) containing acetic acid (0.25 ml), at YC. After stirring for 30 minutes at 50C, the mixture was concentrated to remove the CH,OH and then ethyl acetate 50 (20 ml) was added. The ethyl acetate solution was washed with H20 (2x5 ml), saturated aqueous NaHC03 (5 ml) and brine (5 ml), successively and dried over MgSO,. Evaporation of the solvent gave 145 mg (97%) of the title compound XVIII (Z isomer) as a yellow powder. M.p.

> 1 00"C (dec.).

IR 1,KBr cm--I 3400, 1770, 1720.

max UV: AE10H nm (c) 252 (3700), 258 (3800), 260 (4000), 274 max (4000), 285 (4000). 60 Preparation No. 9 Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-(3-chloro-1-p ro- penl-yl)-3-cephem-4-carboxylate (XVIII- 1) 54 GB2194790A 54 A mixture of 2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetic acid (111-1) (10.1 g, 50 mmole) and PC[, (10.4 g, 50 mmole) in dry CH2CI2 0 00 M0 was stirred at - 7 to - 15'C for 2 hours. The clear solution was poured into n-hexane (500 ml) to give a precipitate. The organic layer was discarded by decantation and the remaining solid was triturated with n-hexane (100 ml). The yellow precipitate was collected by filtration and dried in vacuo to give 12.5 g (99%) of the acid chloride, melting at 800C (dec.).

IR: Vniijol CM-1 1770.

max - The acid chloride (25 mg, 0.1 mmole) was added to a solution of XVIII (Z isomer) from Preparation No. 8 (44 mg, 0.1 mmole) in dry CH,C12 (5 mi) at room temperature, with stirring. After 30 minutes, the mixture was concentrated under reduced pressure and diluted with ethyl acetate (20 m]) and saturated aqueous NaHCO, (5 mi). The organic layer was washed with saturated aqueous NaHC03 (5 m]), brine (5 mi), 10% HCI (5 mi) and brine (5 mi). The solvent was dried over M9SO, and then evaporated to dryness to give the product as a yellow foam. The foam was purified by silica gel (Wakogel C-200, 1 g, containing 0.1 mi of 1/1.5 M phosphate buffer pH 6.4) column chromatography by elution with CH2C12-CH301-1 (100:1), to give 31 mg (50%) of the title compound V111-1 (Z isomer) as a yellow powder. M.p. >l5WC (dec.).

IR: vKBr cm-1 3400, 1775, 1720, 1675, 1630.

max UV: ACOH nm (e) 240 (17000), 280 (10000).

max NIVIR: jMSO-d6 3.6 (21-1, m, 2-1-1), 3.92 (3H, s, O-CH,), 4.0 (21-1, PPM m, CHCl), 5.27 (21-1, m, 6-H & 3-CH=CM, 5.83 (11-1, d-d, J=4.5 & 10, 7-H), 6.25 (1 H, d, J= 11, 3-CM, 30 6.83 (1H,s, CHPhA, 7.33 (10H, m, Ph-H), 8.0 (2H, br-s, NH2), 9.57 (1H, cl, J=10, 7-NH).

- Preparation No. 10 DiphenyImethyl 7[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-[3-iodo-l-pr open- 35 l-yl)3-cephem-4-carboxylate (IX- 1) A solution of V111-1 from Preparation No. 5 (Z/E=2/1, 480 mg, 0.77 mmole) in dry acetone (10 mi) containing Nal (346 mg, 2.3 mmole) was stirred for 30 minutes at ambient temperature.

The reaction mixture was evaporated under reduced pressure. The resulting oil was partitioned between ethyl acetate (50 mi) and water (10 mi). The upper layer was washed with 10% w/v 40 aqueous sodium thiosulfate solution (10 m]) and brine (10 mi) successively, and dried over MgSQ,. Evaporation of the solvent gave 540 mg (98%) of the title compound IX-1 (Z/E=1/1) as a reddish amorphous solid, melting at >l2WC (dec.).

IR VKB, cm-1 3300, 1780, 1720, 1680, 1620.

max UV: AEIOH nm (8) 240 (21000), 290 (12000). max 50 NMR: JDMSO D203.67 (21-1, m, 2H), 5.29 (1H, d, J=4.5, 6-H), PPM 5.95 (1H, d, J=4.5, 7-H), 6.27 (1/2H, d, J = 11, 3-CH cis), 6.72 (1/21-1, d, J = 16, 3-CH trans), 6.87 & 6.96 (each 1/2H, s, ChPh2), 7.4 (1 OH, m, Ph-H). 55 Preparation No. 11 Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido1-3-(3-iodo-1-pr open- l-yl)-3-cephem-4-carboxylate (IX- 1) A mixture of V111-1 (Z isomer) from Preparation No. 9 (5.6 9, 9 mmole) and Nal (4 g, 27 60 mmole) in dry acetone (100 mi) was stirred for 1.5 hours at room temperature. The mixture was evaporated and the resulting oil was diluted with ethyl acetate (90 m]). The ethyl acetate layer was washed with 10% w/v aqueous sodium thiosulfate solution (10 m]) and H20 (10 M1), Removal of the dried (MgSOJ solvent gave a yellow oil, which was solidified by trituration with isopropyl ether. Filtration of the precipitate gave 4.3 9 (67%) of the title compound IX-1 as the GB2194790A 55 E isomer. M.p. > 1WC (dec.).

IR: VKBr cm-1 3400, 1780, 1725, 1680, 1610.

max UV: ROH nm (e) 240 (18000), 297 (11000).

max NW:JMSO-d6+D20 3.90 (3H, s, OCI-1j, 5.25 (1 H, m, 6-H), 5.95 ppm (11-1, m, 7-1-1), 6.72 (d, J=16, 3-CH trans), 6.96 10 (1H, s, CH-Ph2), 7.4 (10H, m, Ph-H).

Preparation No. 12 Benzhydryl 7-Amino-3-[3-chlora-1-propen-l-yll-3-cephem4-carboxylate (Z-isomer) (XVI/1) Compound XVIII is the common intermediate utilized in Reaction Schemes 1b and lc.

A. Benzhydryl 7-BenzylideneMino-3-triphenylphosphoniomethyl-3-cephem-4carboxylate chloride (XV) To a suspension of benzhydryl 7-amino-3-chloromethyi-3-cephem-4- carboxylate hydrochloride (11 hydrochloride) (200 9, 0.44 mole) in CH2C12 (940 m]) was added 1 N NaOH (440 m]) at room 20 temperature. The mixture was shaken for 10 minutes and the organic layer was separated. To the organic layer were added M9SO, (75 9) and benzaidehyde (51 9, 0.48 mole) and the mixtue was allowed to stand for 3 hours. The reaction mixture was filtered and the insolubles were washed with CH2C12 (200 mi). To the combined filtrate and washings was added triphenylphos phine (126 9, 0.48 mole). The mixture was concentrated to about 400 mi and allowed to stand 25 for 4 days. The resulting viscous oil was diluted with ethyl acetate (1 L) and triturated to separate the title compound XV a pale yellow crystalline powder which was collected by filtration and dried in vacuo. Yield 322 9 (96%). M.p. 185-190'C (dec.).

IR VKBr CM - 1 1780, 1720, 1630. 30 max UV ACH2CI2 nm (8) 260 (24100).

niax B. Benzhydryl 7-Benzyfideneamino-3-[(triphenylphosphoranylidene)methyll-3cephem-4-carboxy late - (X VO A mixture of XV (322 9, 0.42 mole), and 5 N Na2C03 (252 mi) in CH2C12 (1. 6 L) was stirred vigorously for 15 minutes at room temperature. The organic layer was separated, dried over M9S04 and concentrated to about 500 mi of volume. The concentrate was added 40 to acetone (1 L), with stirring, to give a light yellow crystalline powder which was collected by filtration to yield 237 9 (78%) of XVI, melting at 195-198'C (dec.).

IR VKBr CM 1 1770, 1620.

max UV: CHc'2 nm (e) 254 (23000), 389 (22000).

Max NIVIR,5MC13 2.56&3.16 (21-1, ABq), 5.00 (11-1, d,J=4 Hz), 5.23 PPM (11-1, d,J=4Hz), 5.47 (1H, cl,J=22 Hz), 6.95 (11-1, s), 7.2-7.8 (301-1, m), 8.55 (11-1, s).

C. Benzhydryl 7-Amino-3-[chloro-l-propen-l-yll-3-cephem-4-carboxylate hyd S ochloride (Z isomer) (XVIII Hydrochloride) To a refluxing solution of W (214 9, 0.294 mole) and N,O-bis- (trimethyisiiyi)acetamide (40 mi, 0.15 mole) in dry CH,C12 (2.9 L) was added dropwide, with stirring, a 50% solution of chloroacetaidehyde (93 9, 0.59 mole) in CI-ICI, over a period of 15 minutes. After standing for minutes, the mixture was concentrated to dryness. To the residual oil were added CH2C12 (1.5 L), Girard Reagent T (99 g, 0.59 mole) and 10% aqueous HCI (300 mi), and'the mixture 60 was stirred for 1 hour at room temperature. The organic layer was washed with water (200 m]) and a saturated NaCI solution (200 ml), dried over M9SO,, treated with charcoal (5 g) and filtered. The filtrate was cooled to - 1 O'C and treated with 1 N HCl in CH30H (300 mi). The mixture was stirred for 30 minutes at room temperature and concentrated to about 300 m]. The concentrate was diluted with ethyl acetate (400 mi) and seeded with a few crystals of XVIII 56 GB2194790A 56 hydrochloride. After 2 hours the separated crystals were collected by filtration, washed with ethyl acetate (200 m]) and dried in vacuo to give 74 9 (53%) of the title compound XVIII as its hydrochloride, melting as > l8WC (dec.). Pale yellow needles.

IR VKBr cm-1 2830, 1780, 1720.

max UV: ZEtOH nm (e) 286 (8800).

max 10 NMR: JOMS0d6 3.73 (2H, br, s, 2-H), 3.97 (2H, m, CHCl), 5.22 ppm (11H, cl, J=4.5 Hz, 6-H), 5.37 (1H, cl, J=4.5 Hz, 7-H),5.77(1H,m,3-CH=CM,6.45(1H,d,J=11 Hz, 3-CM, 6.88 (11H, s, CliPh2), 7.33 (10H, br, s, Ph-M. 15 Anal. Cale'd for C23H211M203SCI. HCI: C, 57.87; H, 4.65; IM, 5.87; S, 6. 72; Cl, 14.85.

Found: C, 57.62; H, 4.53; N, 5.70; S, 6.64; Cl, 14.89.

Preparation No. 13 Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidol3-[3-chforo-1-p ropen-1- yll-3-cephem-4-carboxylate j isomer) (VIII- 1) To a stirred solution of XVIII (Z isomer) (20 g, 42 mmole) in CH2C12 (420 m]) containing N,O bis(trimethyisiiyi)-acetamide (34 mi, 125 mmole) was added 2-(5-amino-1,2, 4-thiadiazol-3-yi)-2- methoxyiminoacetyl chloride hydrochloride (15.2 9, 59 mmole) in three portions over a period of 25 minutes at -10 to O'C. The mixture was stirred for 30 minutes at O-WC and concentrated under reduced pressure. The residual brown oil was dissolved in ethyl acetate (420 mi) and the solution was washed successively with saturated aqueous NalHIC03 (3 x 15 mi), saturated aque ous NaCI (15 mi), 10% HCI (15 mi) and saturated aqueous NaCI (15 m]), and concentrated to about 50 mi of the volume. To the concentrate was added n-heptane (200 mi) to give 28.5 9 30 (90% pure) of the title compound V111-1 (Z-isomer) as a colorless powder. M.p. >l5WC (dec.).

: VKBr cml 3400, 1780, 1720, 1680, 1620.

max UV: AEtOH nm (e) 240 (20000), 283 (12000).

max NIMR: jacetone-d. 3.6 (2H, m, 2-H), 3.95 (3H, s, OCH,), 4.0 PPM (2H, m, CHCl), 5.32 (111, cl, J=4.5 Hz, 40 6-H), 5.62 (1H, m, 3-CH=CM, 6.03 (1H, cl, J = 4.5 Hz, 7-H), 6.32 (1 H, cl, J = 11 Hz, 3-CH), 6.87 (1H, s, ChPh2), 7.33 (10H, br, s, Ph-H).

Preparation No. 14 Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamidol-3-[3-iodo-l-pr open-lE yll-3-cephem-4-carboxylate E isomer) (IX- 1) A mixture of V111-1 (Z isomer) (28.5 9, 90% pure) and sodium iodide (19 9) in dry acetone (420 mi) was stirred for 10 minutes at room temperature and allowed to stand at WC for 2 hours. The mixture was concentrated under reduced pressure. To the residue was added ethyl 50 acetate (420 m]) and 10% w/v aqueous sodium thiosulfate solution (30 mi), and the mixture was shaken. The organic layer was washed with water (30 ml), dried over MgSO, and evapo rated to about 50 mi of volume. The concentrate was diluted with n- heptane (200 mi) to yield 30.6 g (95% pure) of the title compound IX-1 (E isomer) as a yellow powder, melting at >l2WC (dec.).

IR VKBr cm-1 3400, 1780, 1725, 1680, 1620.

max UV: AROH nm (e) 306 (15000).

max 57 GB2194790A 57 NMR: 5acetone-d6 3.71 (2H, m, 2-H), 3.97 (3H, s, OCH3),4.0 PPM (2H, cl, J=8 Hz, CH21), 5.26 (1H, cl,J=4.5 Hz, 6-H), 6.03 (1 H, d-d, J=4.5 & 8 Hz, changed to doublet J=4.5 Hz by D,O, 7-H), 6.32 (1H, d-t, J = 15 & 8 Hz, 3-CH = CH, 6.79 (1 H, d, J = 15 HZ, 5 3-CM, 6.98 (1H, s, CHPh,), 7.35 (10H, m, Ph-H), 7.63 (2H, br, s, disappeared by D,O, NH2), 8.52 (1H, d, J=8 Hz, disappeared by D,O, 7-NM.

10 Preparation No. 15 Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetamidol3-[3-(4-carbam oyl-l-py- ridinio)-1-propen-l-yll-3-cephem-4-carboxylate Iodide (E isomer) (XII-1H) To a suspension of IX-1 (E isomer) (30.5 9) and isonicotinamide (26 9, 212 mmole) in CH.M (120 mi) was added CH30H (100 mi) until the mixture became clear. The solution was stirred for 15 2 hours under nitrogen atmosphere at room temperature and concentrated to about 100 mI under reduced pressure. The residual semi-solid was triturated with isopropyl ether (200 m[).

The solvent was removed by decantation and the residual yellow powder was washed with a mixture of isopropyl ether and CH,OH (3/1, 120 m[). The powder was collected by filtration and dried in vacuo to give 36 9 (75% pure estimated by HPLC) of the title compound X11-11-1 (E 20 isomer) as a light yellow powder melting at >l5WC (dec.).

IR VKBr cm-1 3300, 1780, 1720, 1680, 1620.

max UV: AROH nm (EI-AH) 282 (170).

max 1 Cm NMR: JWSOd6 3.72 (2H, m, 2-H), 3.90 (3H, s, OCH,), 5.25 (3H, PPM rn, 6-H & CHN1), 5.9 (1H, d-d, J=4.5 & 8 Hz, 30 changed to a doublet J=4.5 Hz by D20 addition, 7-H), 6.35 (1H, rn, 3-CH=CM, 6.89 (1H, s, CHPhA, 6.9 (1H, d, J=16 Hz, 3-CiA, 7.35 (10H, rn, Ph-H), 8.06 (2H, br, s, disappeared by D201 NH2), 8.21 (2H, br, s, disappeared by D20 35 addition, NH2), 8.36 & 9.07 (each 2H, d, J6 Hz, Py-M, 9.57 (1H, d, J=8 Hz, disappeared by D20 addition, 7-NH).

1 45 Preparation No. 16 Benzhydryl 7-Benzylideneamino-3-[3-chloro-l-propen-1-yll-3-cephem-4- carboxylate (XV//) (Z isomer) To an ice-cooled mixture of the crystalline 7-amino-cephem intermediate XVIll (Z isomer) (13.4 g, 28 mmole) and benzaldehyde (3.3 g, 31 mmole) in ethyl acetate (150 ml) was added dropwise 0.5 N sodium hydroxide (56 mi, 28 mmole) over a period of 20 minutes, to maintain the temperature of the reaction mixture below 10'C. The mixture was stirred with cooling for another 15 minutes, and the organic layer was separated, washed with saturated aqueous sodium bicarbonate (100 mlx2) and dried over magnesium sulfate. To the dried solution was added a small amount of charcoal and the mixture was filtered. The filtrate was concentrated to dryness. The residual oil was dissolved in carbon tetrachloride (50 ml), and concentrated again. 50 This procedure was repeated 3 times, and the mixture was monitored by reverse phase tlc to confirm that all of the starting 7-amino-cephalosporin was converted to the Schiff's base.

Removing the solvent in vacuo gave 16.45 g of the title compound XVII (Z isomer) as a pale yellow powder (estimated purity 85%; M.p. 74'C (dec.), which was used for the next step without purification.

IR VKBr CM-1 1780, 1725, 1635.

max UV:ACH2c'2nm (E'%) 257 (400).

max 1 cm NMR: 5CDC13 6,18 (1 H, cl, J = 11 Hz).

PPM 40.

58 GB2194790A 58 Preparation No. 17 Benzhydryl 7-Benzyiideneamino-3-[3-(4-carbamoyl-l-pyridinio)-1-propen-1yll-3-cephem-4-c arboxyl- ate Iodide (XXl-H) (E isomer) To a chilled mixture of the 3-chloropropenylcephem XVII (Z isomer) (16.4 9) in acetone (5 mi), was added dropwise a solution of sodium iodide (6.3 9, 42 mmole) in acetone (30 mi) over 10 5 minutes under nitrogen atmosphere, and the mixture was stirred at room temperature.

The reaction was monitored by the ratio of uv absorption [E] % (255 nm)/Ell% (320 nm)l.

em c When the ratio reached below 1.30 (after 45 minutes), the mixture was diluted with carbon tetrachloride (400 mi), and allowed to stand at room temperature. When the ratio came to below 1.10 (after 3 hours), the mixture was concentrated to a half its volume. The concentrate 10 was treated with a small amount of charcoal and diatomaceous earth, and filtered. The filter cake was washed with a 1:1 mixture (100 mi) of methylene chloride and carbon tetrachloride.

To the combined solution of the filtrate and washings, was added a solution of isonicotinamide (1.5 g, 28.7 mmole) in dimethylformamide (20 mi) and the mixture was concentrated under reduced pressure. The concentrate was allowed to stand at room temperature for 1.5 hours and 15 washed with isopropyl ether (100 m[x3). The residual brown semi-solid was dissolved in methylene chloride (50 m]) and the solution was added dropwise, with stirring, to ethyl acetate (1.5 L). The resulting precipitate was collected by filtration and washed with ethyl acetate (200 mi). After drying over phosphorous pentoxide in vacuo, 17 9 of the title compound M-H (E isomer) was obtained. Yellow amorphous powder. M.p. 1150-155'C (dec.). Estimated purity 80% 20 by nmr.

IR VKBr CM-1 1775, 1725, 1690, 1635.

max UV:),CH2c' nm (E'%) 258 (335, 298 (255).

max 1 CM NMR: jDfASO-d. 3.4-3.8 (211, br.), 5.35 (2H, br.), 5.41 (1H, d, PPM J=4 Hz), 5.73 (1H, d, J=4 Hz), 6.93 (1H, s), 6.97 (1H, cl, J=16 Hz), 7.3-7.5 (15H, br. s), 8.40 (2H, d, J=6.5 Hz), 9.15 (2H, d, J=6.5 Hz). Preparation No. 18 7-Amino-3-[3-(4-carbamoyl- 1-pyridinio)- 1-propen-

l-yll-3-cephem-4- carboxylate (XXII-H) (E isomer) To a suspension of the quaternized cephem XXl-H (17 g) in 85% formic acid (25 mi) was added dropwise concentrated hydrochloric acid (5 mi), and the mixture was stirred at room temperature for 1.5 hours and treated with a small amount of charcoal. The mixture was filtered and washed with 85% formic acid (5 mi). The filtrate was combined with the wash and poured 40 into acetone (1 L), with stirring. The resulting precipitate was collected by filtration to give 9.52 9 of yellow-colored crude product. To a suspension of the crude material (9.5 9) in water (50 m]) was added a small amount of charcoal, and the mixture was filtered. The filtrate was added dropwise, with stirring, to isopropyl alcohol (700 mi). The resulting precipitate was collected by filtration, washed with a small amount of methanol (30 mi), and dried to give 7.58 9 of the title 45 compound MI-Fl (E isomer) as the hydrochloride. Light yellow powder. Estimated purity 85% by UV. M.p. 1173-188'C (dec.).

IR VKBr CM 1 1795, 1680, 1620, 1575, 1540.

max uv: A Phosphate buffer (pH 7) nm (E'%) 294 (457).

max 1 cm NMR: 3D20+M 3.82 (2H, s), 5.17 (1H, cl, J=5 Hz), 5.33 (2H, ppm cl, J=7 Hz), 5.43 (1H, cl, J=5 Hz), 6.37 (1H, d-t, J=16 & 7 Hz), 7.23 (1H, d, J=16 Hz), 8.34 (2H, d, J=7 Hz), 9.00 (2H, cl, J=7 Hz).

Preparation No. 19 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetyI Chloride Hydrochloride (111-1 as its acid chloride hydrochloride) A. 2Cyano-2-methoxyiminoacetamide To a stirred mixture of a-cyanoacetamide (252 9, 3 mole) and sodium nitrite (414 9, 6 mole) in water (600 m]) was added acetic acid (371 mi, 10 mole) at 5-10'C over 1.5 hours. The 65 59 GB2194790A 59 mixture was allowed to stir for another 1.5 hours and adjusted to pH 8.5 with 6 N NaOH. To the mixture was added dimethyl suffate (568 m], 6 mole) at 1520'C and the mixture was stirred at 4WC for 1.5 hours. The reaction mixture was adjusted to pH 8.5 with 6 N NaOH and allowed to stand at WC overnight to separate the precipitate, which was collected by filtration, washed with cold water and air-dried to give 292 g (77%) of the title compound as brown needles melting at 170-1720C.

IR: VKBr cm-1 3400, 3180, 1720(sh), 1715, 1690, 1615, 1570.

max UV: AH20 nm (g) 238.5 (8290), 268 (sh, 3870).

max NIVIR: 6MSO-d6 4.20 (3H, s, OCH,), 7.85 (2H, br. NH2).

PPM Anal. Calc'cl. for C^N302: Found:

B. 2-Methoxyiminopropanedinitrile C, 37.80; H, 3.97; N, 33.06 C, 37.43; H, 3.75; N, 32.51.

A stirred mixture of 2-cyano-2-methoxyiminoacetamide (88.9 g, 0.7 mole), sodium chloride (70 g) and phosphorus oxychloride (97 ml, 1.05 mole) in dry 1,2-dichloroethane (350 ml) was refluxed for 16 hours. The insolubles were filtered off through a dicalite pad and washed with dichloroethane. The filtrate and the wash were combined, and poured into stirred icewater (1.5 L) to decompose the excess of phosphorus oxychloride. The organic phase was washed with 10% NaHC03 (500 ml), water (500 mlx3) and a saturated NaCl solution (500 ml), and dried over MgSO,. The filtrate was distilled under diminished pressure to give 61.5 g (81%) of the title compound boiling at 62'C/24 mm Hg. (Lit., b.p. 47-48'C/12 mm Hg).

IR VUquidFilm cm-1 3020, 2960, 22451 2020 1530, 1455, 1080.

max NIVIR: JCDcl., 4.35 (3H, s, OCH3).

PPM C. 2-Cyano-2-methoxyiminoacetamidinium Acetate To a solution of ammonium chloride (28.4 g 0.53 mole) in 28% aqueous ammonia (355 ml) and ethanol (180 ml) was added dropwise a solution of 2methoxyiminopropanedinitrile (58.0 g, 0.53 mole) in ethanol (120 ml) at 15 to - 1 OC over a period of 30 minutes, with stirring.

The mixture was stirred at - 1 O'C overnight and then at ambient temperature (20-25C) for one 40 day. The reaction mixture was partitioned between water (350 ml) and CH2CI2 (350 ml), and the aqueous phase was saturated with sodium chloride, and extracted again with CH2CI2 (300 ml).

The organic extracts were combined, dried over MgSO, and evaporated in vacuo. A solution of the residue in ethyl acetate (1.6 Q was adjusted to pH 3-4 with acetic acid to precipitate the title compound as crystals, which were collected by filtration and washed with ethyl acetate. 45 Yield 67.6 g (69%), M.p. 152-4'C (dec.) [Lit., m.p. 150-155'C (dec).].

PPM IR VKBr cm--I 3160, 2900, 2360, 2235, 2000, 1665, 1555, max 195, 1415.

UV: AE10H nm (e) 243 (8500), 265 (sh, 5380), 305 (sh, 1400).

-ax NIVIR: JWSO-da 1.88 (3H, s, CH3COOH), 4.15 (3H, s, OCH3), 7.60 (4H, br.).

Anal. Calc'd for C4H6N,O. CH3COOH: C, 3 8.7 Found: C, 38.7 1; 1; H, 5.41; N, 30.09 H, 5.59; N, 29.51 D. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetonitrile To a suspension of 2-cyano-2-methoxyiminoacetamidinium acetate (125 g, 0. 672 mole) in CH30H (1.25 L) were added dropwise triethylamine (234 m], 1. 68 mole) at - 1WC, and subsequently Br, (41.6 mi, 0.806 mole) over 20 minutes at -15 to -10'C, and the mixture was stirred for 20 minutes. To the mixture was added dropwise a solution of KSM (78.3 9, 0.806 mole) in CH30H (550 ml) over 1 hour at - 15 to - 1 O'C. After stirring at O-WC for 1 hour, the GB2194790A 60 mixture was poured into ice-water (12 L) to form a crystalline precipitate, which was collected by filtration, washed with water and air- dried to give 120 9 (98%) of the title compound. M.p. 263-WC (dec.). The m.p. of the compound prepared by us is higher by about WC than that given in the literature [m.p. 210-15'C (dec.)], but our spectral and microanalytical data are 5 consistent for the structure.

IR VKBr cm-1 3435, 3260, 3120, 2960, 2245, 2020, 1630, max 1545, 1455, 1415.

UV:AEtOH nm (e) 248 (13300), 310 (3470).

max NMR: 5DMSO-d. 4.21 (3H, s, OCI-1J, 8.30 (21-1, br. NH2).

PPM Anal. Calc'd for C.1---1^OS: C, 32.78; H, 2.75; N, 38.23; S, 17.50 Found: C, 32.76;H, 2.51; N, 38.02; S, 17.50.

E. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetic Acid (111-1) A mixture of 2-(5-amino-1,2,4thiadiazol-3-yl-2-methoxyiminoacetonitrile (18.3 9, 0.1 mole) in 4 20 N NaOH (250 mi) was heated at 50-55'C with stirring for 3 hours. The reaction mixture was adjusted to pH 1 with H,PO,, and washed with ethyl acetate (100 mi), saturated with NaCI, and extracted three times with a mixture of ethyl acetate and tetrahydrofuran (3.1, 300 mlx2, and mix 1). The extracts were combined, dried over M9SO, and concentrated under reduced pressure. The residue was triturated with isopropyl ether to afford pale yellow crystals of the title acid. Yield 16.8 9 (83%). M.p. 184-WC (dec.). [Lit.. m.p. 180-182'C (dec.)].

IR: VKBr cm-1 3460, 3260, 3140, 1725, 1620, 1605, 1546.

max UV: AH20 nm (E) 234 (13200), 288 (sh, 3620).

max F. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetyl Chloride Hydrochloride To a suspension of 2-(5-amino-1,2,4-thiadiazol-3-yi)-2-methoxyiminoacetic acid (111-1) (40.4 9, 35 0.2 mole) in dry CH2C12 (400 mi) was added PC], (41.6 g, 0.2 mole) in one portion at -500C.

The mixture was stirred for 4 hours at -20 to -WC, and poured into a mixture of n-heptane and isopropyl ether (2A, 2 L). The yellow precipitate was collected by filtration, washed with the same solvent mixture, and dried with KOH under reduced pressure to give 46.0 9 (90%) of the title acid chloride.

M VNLIjol CM-1 1775.

max Preparation No. 20 N C-CONH 5 11 1 H2N kS,.A 9\ 0 V111-2 z -1 1 C2H5 o- k. CH2CH-CH2-CI COOMPM2 Z 5Q DiphenyImethyl 7-[2-(5-Amino1,2,4-thiadiazol-3-yl)-2-(Z)- ethoxyiminoacetamidol-3-[3-chloro- 1-pro penyll-3-cephem-4-carboxylate (V111-2, Z isomer) To a mixture of N,O-bis(trimethyisiiyi)acetamide (2.3 mi, 9 mmoles) and crystalline diphenyime thyl 7-amino-3-[3-chloro-l-(Z)-propen-l-yil-3-cephem-4-carboxylate hydrochloride (XVIII) (1.338 9, 2.8 mmoles) (from Preparation No. 12) in methylene chloride (10 mi) was added 2-(5-amino 1,2,4-thiadiazol-3-y])-2-(Z)-ethoxyiminoacetyl chloride hydrochloride (800 mg, 2.95 mmoles) por tionwtse, with stirring, at -10'C and the mixture was allowed to stand at O'C for 2 hours. The 60 mixture was diluted with ethyl acetate (200 ml), washed with water and evaporated under reduced pressure. Trituration of the residue with isopropyl ether afforded the title product V111-2 as an amorphous powder. Yield 1.70 9 (95%) Mp. >1500C (dec.).

Japan Kokai 57-158769 published September 30, 1982, to Fujisawa (Brit. appl., 3/6/81).

61 GB2194790A 61 IR: v (KBr) in cm-1 3300, 1780, 1720, 1690, 1380, 1220.

UV: (C^OH) in nm (e) 285 (11000).

NIVIR:.5 (DIVISO-dj in ppm 1.26 (3H, t, J=7Hz, CH2CHJ, 4.25 (2H, q, J=7Hz, CH2CH3), 5.90 (1H, d-d, J=4 & 8Hz, 7-H), 6.26 (1H, cl, J=111-1z, 3-CH), 6.85 (1H, s, CHPh2), 9.53 (1H, d, J=81-1z, 7-NH).

Preparation No. 21 H C - CONN - 11 11 S"N N\0 H 49 N2NJ P CH=CH-C 1 1 0 C2H5 CODCH(M2 IX-2 A mixture of E and Z isomers Diphenyimethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(Z)ethoxyiminoacetamidol-3-[3-iodo-l -propenyll-3-cephem-4-carboxylate (IX2) A mixture of V111-2 (1.90 9, 3 mmoles) (from Preparation No. 20) and sodium iodide (1.4 9, 9 mmoles) in acetone (20 mi) was stirred for 10 minutes at room temperature and then allowed to stand at 5'C for 3 hours. The mixture was evaporated under reduced pressure, diluted with ethyl 25 acetate (100 mi), washed with 10% sodium thiosulfate and water, and evaporated under re- duced pressure. Trituration of the residue with isopropyl ether gave 1.82 9 (84%) of the title - product IX-2 as a light brown amorphous powder.

IR: v (KBr) in cm-1 3290, 1770, 1720, 1670, 1530, 1370, 1220. 30 UV (C21-1,01-1) in nm (E"') 304 (199).

max Preparation No. 22 1 CM n-CH= N - N -CONH2 COO CH (M2 M-H iodide E Diphenyimethyl 7-Benzylideneamino-3-[(E)-3-(4-carbamoylpyridinio)-1propenyll-3-cophem-4-ca r- 45 boxylate (XXl-H iodide) (E isomer) To a chilled solution of the 3-chloropropenylcephem (XVII, Z isomer, 42.8 9, 90 mmolesI (from Preparation No. 16) in dry DMF (80 mi), was added KI (20 9, 120 mmoles) in one portion, and the mixture was stirred at room temperature.

The reaction was monitored by the ratio of UV absorption [E'% (255 nm)/E1% (320 nm)]. 50 1 CM 1 CM When the ratio became below 1.10 (after 45 minutes), the mixture was diluted with 800 m] of methylene chloride, treated with active carbon (4 g), and filtered. The filter cake was washed with 100 mi of CH2C12. To the combined filtrate and washings was added isonicotinamide (14.64 g), and the mixture was concentrated under reduced pressure. The concentrate was kept at room temperature for 1.5 hours and washed with a mixture of toluene and n-heptane (1A, 600 m]). The residual brown semi- solid was dissolved in CH2C12 (100 mi) and the solution was added dropwise to ethyl acetate (3 L) with vigorous stirring. After drying over P,O, in vacuo, 57.37 g (88%) of the quaternized title product XXI-H was obtained as the iodide. Yellow amorphous powder. Mp. 150-155'C (dec.). This product was identical to that obtained by iodination with Nal (Preparation No. 17).

62 GB2194790A 62 Preparation No. 23 KI'N2N 5 0 CH-CH- CH2-Cl coo CH(Fhh xVill Z Diphenylmethyl 7-Amino-3-(3-chloro-1-propenyl)-3-cephem-4-carboxylate hydrochloride (Z isOMer) (XVIII, Hydrochloride) A 25% solution of chloroacetaidehyde (69 g, 0.22 mmoles) in CHCI, was added to a solution of W (80 g, 0.11 mole) in CH,Cl, (1.1 L) containing N,O- bis(trimethyisiiyi)acetamide (16.2 mi, 0.06 mole) at -10'C in one portion, and the mixture was allowed to stand overnight at WC.

The mixture was concentrated to ca. 0.3 IL, diluted with a mixed solvent of ethyl acetate and isopropyl ether (l/2, 0.6 L), treated with silica gel (Wakogel C-100, 60 g) and filtered through a dicalite pad. The filter cake was washed with the same solvent system (0. 2 L). The combined filtrate and washing were concentrated to ca. 0.2 L, treated with Girard Reagent T (60 9, 0.26 mole) and 4N HCI (220 mi), and seeded with a few crystals of XVIII hydrochloride. After stirring 20 for 3 hours, the resulting crystals were collected by filtration, washed with water (0.5 L) and ethyl acetate (0.5 L) and dried in vacuo to give 37 g (70%) of the title compound XVIII hydrochloride, melting at > 18WC (dec.). Pale yellow needles. This product was identical to that obtained in Preparation No. 12.

Preparation No. 24 MCH2N- 5 CH=CH- CH2-Cl [00 CH(Phh XVIII z DiphenyImethyl 7-Amino-3-(3-chloro-1-propenyl)-3-cephem-4-carboxylate hydrochloride (Z isomer) 35 (XVIII, Hydrochloride) To a solution of chloroacetaldehyde (25% solution in CHC13, 628 mg, 2 mmoles) in CH2C12 (10 mi) were added N,O-bis-(trimethyisiiyi)acetamide (0.135 mi, 0.5 mmole) and W (728 mg, 1 mmole), successively, at WC. The mixture was allowed to stand overnight at WC. The mixture was evaporated and diluted with a mixture of ethyl acetate and isopropyl ether (l/2, 10 mi). Insolubles were removed by filtration and the filtrate was concentrated to ca. 5 m]. The concentrate was treated with 4N 1-10 (2 mi), seeded with XVIII hydrochloride and stirred for 1 hour at room temperature. The crystals were collected by filtration, washed with ethyl acetate (10 mi) and water (10 mi) and dried in vacuo to give 384 mg (80%) of the title compound XVIII hydrochloride, melting at > 18WC (dec.). Pale yellow needles.. This product was identical to that obtained by Preparation No. 12.

Preparation No. 25 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)acetyl chloride hydrochloride (111-3 as its acid chloride hydrochloride A. Methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-(propen-3yloxyimino)ace tate A mixture of 685 mg (3.37 mmoles) of W(propen-3-yloxy)phthalimide [prepared according to the procedure of E. Grochosaki & J. Jurczak, Synthesis 1976 682] and 175 mg (3.35 mmoles) of hydrazine hydrate in 5 m] of C^OH was stirred for 1 hour at room temperature. The resulting precipitate was filtered off and the filtrate and washings were combined. To the 55 solution was added 967 mg (3.37 mmoles) of methyl 2-(5-t- butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-oxoacetate, and the mixture was allowed to stand for 1 hour at room temperature and concentrated by a rotary evaporator. The residue was purified by silica gel chromatography.

The column was eluted with n-hexane/ethyl acetate (4:1) and fractions containing the major product were combined and evaporated under reduced pressure. Yield 514 mg (46%). Mp. 60 83-86'C.

IR: v (KBr) in cm-1 3100, 1745, 1710, 1610.

63 GB2194790A 63 UV: A (C2H,OH) in nm (e) 223 (9700), 242 (10000).

NMR: J (C13C13) in ppm 1.55 (9H, s, BOC-1-1), 4.40 (2H, cl, J=5Hz, O-CH2), 5.21 (2H, m, CH2=CH).- 5.90 (11-1, m, -CH=CH2), 9.50 (11-1, br.s, NH).

B. 2-(5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-(propen3yloxyimino)ace tic acid(') A solution of 770 mg (2.3 mmoles) of methyl 2(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3yi)-2-(propen3yioxyimino)acetate and 3.5 mi of 2N NaOH solution (7.0 mmoles) in 15 mI of CH,OH was refluxed for 30 minutes. The reaction mixture was concentrated in vacuo and diluted 10 with 10 mi of ethyl acetate-1-1,0 (1:1). The water layer was separated, acidified to pH 2 with 6N HO and extracted with ethyl acetate (10 mIX2). The ethyl acetate solution was dried over M9SO, and concentrated by a rotary evaporator to afford 596 mg (81%) of the title compound. Mp. 134-13WC (lit(l: mp. 135-136'C.

IR: Vmax (Nujol) in cm-1 3150, 1745, 1710, 1550.

UV: Am. (C,H,OH) in nm (e) 223 (11000), 242 (11300).

NMR: ú5 (DMSO-d6) in ppm 1.55 (9H, s, BOC-H), 4.77 (21-1, d, J=5Hz, OCH2), 5.22 (21-1, m, C142=CH), 6.0 (1H, m, CH=CH2).

(1) 1. Csendes, et al., J. Antibiotics, 36, 1020 (1983).

C. 2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)acetic acid (111-3)(') A solution of 570 mg (1.74 mmoles) of 2-(5-t-butoxycarbonylamino-1,2,4- thiadiazol-3-yl)-2(propen-3-yloxyimino) acetic acid in 6 mi of trifluoroacetic acid was allowed to stand for 1 hour at ambient temperature. Evaporation followed by trituration with 30 mi of isopropyl ether gave 30 376 mg (95%) of the title compound. Mp. 1OWC (dec.).

IR: vna>, (Nujol) in em -1 3180, 1710, 1545, 1460.

UV A (C2H,OH) in nm (a) 24,5 (13500).

35 NIVIR: 3 (DMSO-d6) in ppm 4.77 (21-1, d, J=5Hz, O-CH2) 5.20 (21-1, m, CH=CH), 6.0 (1H, m, CH=CH2).

(1) japan Kokai 57-112396 (7/13/82, Fujisawa) Brit. appl. 7935538 (10/12/79).

D. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)acetyI chloride hydrochloride A solution of 350 mg (1.54 mmoles) of 111-3 and 410 mg (1.97 mmoles) of phosphorous pentachloride in dichloromethane (5 ml) was stirred for 1 hour at 25'C. The reaction mixture was poured into 60 ml of n-hexane and the precipitate was filtered off. Yield 323 mg.

IR: v.,,, (Nujol) in cm-1 1765.

Preparation No. 26 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2propargyloxyiminoacetyl chloride hydrochloride (111-4 as its 50 acid chloride hydrochloride) A. Methyl 2-(5-t-Butoxycarbonylamino-1,2,4thiadiazol-3-yl)-2-propargyloxyiminoacetate A suspension of 870 mg (4.32 mmoles) of N-proparglyoxyphthalimide(l) and 200 mg (4.0 mmoles) of hydrazine hydrate in 5 mi of ethanol was stirred at 2WC for 1 hour and filtered. To the combined filtrate and washings was added 1.0 9 (3.86 mmoles) of methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-oxoacetate(2). The solution was allowed to stand for 1 hour and concentrated under reduced pressure. Purification by silica gel chromatography followed by evaporation afforded 319 mg (27%) of the title product. Mp. 72-75'C.

IR: vna,, (KBr) in cm-1.3200, 2380, 1745, 1710, 1610. 60 UV: Amax (C2H5OH) in rim (e) 235 (12200).

64 GB2194790A 64 NIVIR: 3 (DMSO-d.) in ppm 1,56 (91-1, s, BOC-H), 3.55 (1 H, t, J=2Hz, C=CH), 4.85 (2H, d, J=21HIz, -CH2-C=CH), 8.9 (11-1, br.s, NH).

(1) Commercially available, Aldrich.

(2) 1. Csendes et al., J. Antibiotics 36, 1020 (1983).

B. 2-(5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl)2propargyloxyiminoacetic acid A solution of 490 mg (1.4 mmoles) of methyl 2-(5-t-butoxycarbonylamino-1, 2,4-thiadiazol-3yl)-2-propargyloxyiminoacetate and 2.2 m] of 2N aqueous NaOH solution (4. 4 mmoles) in 14 mi 10 of CH,OH was refluxed for 30 minutes. The reaction mixture was concentrated under reduced pressure and 10 mi of ethyl acetate-1-1,0 (1:1) was added to the solution. The separated water layer was acidified to pH 2 with 6N HO and extracted with ethyl acetate (2 x 10 m]). Drying over MgSO, followed by evaporation of the organic layer gave 149 mg (89%) of the title product.

Mp. 13WC (dec).

IR: v,,,,,, (Nujol) in cm-1 3350, 1720, 1670, 1550.

UV: max (C2H,OH) in nm (6) 233 (11500). 20 NIVIR: 6 (DMSO-d6) in ppm 1.55 (91-1, s, BOC-1-1), 3.55 (1 H, t, J=21-1z, C=-CH), 4.89 (21-1, cl, J=21-1z, CH2C-CH), 9.0 (1H, s, NH).

C. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetic acid (1114)(3) A solution of 410 mg (1.26 mmoles) of 2-(5-t-butoxycarbonylamino-1,2,4thiadiazol-3-yi)-2propargyloxyiminoacetic acid in 5 mi of trifluoroacetic acid was allowed to stand for 1 hour at 25'C. Evaporation followed by trituration of the residue with 25 m] of isopropyl ether gave 204 mg (72%) of the title compound. Mp. 156-158'C (dec.).

IR: v_ (Nujol) in cm - 1 3300, 2480, 1730, 16 10.

UV:)max (C2H,OH) in nm (c) 234 (12000).

NMR: J (DMSO-d6) in ppm 3.52 (11-1, t, J=2Hz, C-CH), 4.86 (21-1, d, J=211z, CH,-C-=CH), 8.10 (21-1, br.s, NH2).

(3) Japan Kokai 57-112396 (7/13/82, Fujisawa) Brit. appl. 7935538 (10/12/79).

D. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetyl chloride hydrochloride A mixture of 175 mg (0.07 mmole) of 111-4 and 182 mg (0.88 mmole) of phosphorous pentachloride in dichforomethane (2 mi) was stirred for 1 hour at -WC. The reaction mixture was poured into 30 mI of n-hexane and the precipitate was filtered off. Yield 65 mg (34%).

IR : v,,,. (Nujol) irl cm-1 1770.

Preparation No. 27 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2cyclopentyloxyiminoacetyl chloride hydrochloride (111-5 as its 50 acid chloride hydrochloride) A. Methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2cyclopentyloxyiminoaceta te A suspension of 860 mg (3.7 mmoles) of N-(cyclopentyloxy)phthalimide(l) and 185 mg (3.7 mmoles) of hydrazine hydrate in 5 mi of C,H,OH was stirred for 1 hour at ambient temperature and filtered. The filtrate and washings were combined and added to 1.06 9 (3.7 mmoles) of methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-oxoacetate(2). The solution was allowed 55 to stand for 1 hour at room temperature and concentrated in vacuo. The residue was purified by silica gel column chromatography. Elution with n-hexane-ethyl acetate (4:1) followed by evapora- tion gave the title product. Yield 906 mg (81%). Mp. 115-118'C.

At 60]R: Vmax (KBr) in cm-1 3200, 1745, 1710, 1550. 60 UV: A (C2H5OH) in nm (e) 217 (1800), 252 (7600).

GB2194790A 65 NIVIR: 8 (CDCI,) is ppm 1.51 (9H, s, BOC-H), 1.60 (8H, br.s, H]), 3.88 (31-1, s, OCH3), 4.90 (1 H, H 0)G ' 8.70 (11-1, br.s, NH).

* (1) U.S. Patent 3.971,778 (7/27/76; Glaxo), Brit. appl. 49255 (10/25/72).

(2) 1. Csendes et al., J. Antibiotics 36, 1020 (1983).

B. 2-(5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2cyclopentyloxyiminoaceti c acid A solution of 500 mg (1.34 mmoles) of methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3yi)-2eyclopentyloxyiminoacetate and 2N NaOH solution (2 mi, 4 mmoles) in 15 mi of CH30H was refluxed for 30 minutes. The reaction mixture was evaporated and 10 mi of ethyl acetate- 15 H,0 (1:1) was added to the solution. The water layer was separated, acidified to pH 2 with 6N HCI and extracted with ethyl acetate (10 mix2). The organic layer was washed with brine, dried over M9S04 and concentrated under reduced pressure to give 377 mg (78%) of the title compound. Mp. 18WC (dec.).

IR: Vmax (KBr) in cm-1 3160, 1710, 1550.

UV: Amax (C2H5OH) in nm (e) 238 (13300).

NIVIR: (5 (DMSO) in ppm 1.51 (9H, s, BOC-H), 1.70 (8H, br.s., 30), 4.82 (1 H, M C. 2-(5-Amino- 1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetic acid (111-5, Z isomer)(3) A solution of 348 mg (0.97 mmoles) of 2-(5-tbutoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2cyclopentyloxyiminoacetic acid in 2 mi of trifluoroacetic acid was allowed to stand for 1 hour at room temperature. The reaction mixture was concentrated under reduced pressure. The residue was triturated with 5 mi of isopropyl ether and 10 mi of hexane to give 215 mg (86%) of the title compound. Mp. 162-165'C (dec.) [liffl): Mp. 160-165'C (dec.)].

IR: v,,,, (Nujol) in cm-1 3290, 3200, 1710, 1615, 1600.

UV: Am (C^OH) in nm (e) 238 (1j300).

NIVIR: ú5 (DMSO-d.) in ppm 1.17-2.10 (8H, m), 4.60-4.98 (11-1, m), 8.22 (21-1, s).

(3) Japan Kokai 57-158769 (9/30/82, Fujisawa) Brit. appl. 8107134 (3/6/81).

D. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetyl chloride hydrochloride A solution of 190 mg (0.74 mmole) of 111-5 and 219 mg (1.0 mmole) of phosphorous pentachloride in dichloromethane (5 mi) was stirred for 1 hour at room temperature. The reaction 50 mixture was poured into 50 m] of n-hexane. The resulting precipitate was collected by filtration. Yield 122 mg (60%).

IR: v,,,, (Nujol) in cm-1 1760.

Preparation No. 28 Benzotriazol- l-yi-2-(5-amino1,2,4-thiadiazol-3-yl)-2- methoxyiminoacetate A mixture of 1-hydroxybenzotriazole (2.7 9, 20 mmoles) and dicyclohexylcarbodiimide (4.12 g, mmoles) in 65 mi of DIVIF was stirred at room temperature. After 15 minutes, 111-1 (4.04 9, 20 mmoles) was added to the stirring mixture at WC, and stirring was continued for 3 hours. 60 The reaction mixture was filtered to remove the insoluble urea, and the filter cake was washed with a small volume of DW The filtrate and washings were combined and poured into 800 mi of ice water. The precipitate was collected by filtration to give 5.24 g (82%) of the title compound as a light grey powder. Mp. 189-192'C (dec.).

66 GB2194790A 66 iR: v,,,,, (KBr) in cm-1 1815, 1620, 1540, 1415, 1090, 1060, 1005, 945, 865, 740.

UV (C2H,,OH) in nm (E",6) 246 (580), 283sh, (228).

max

Claims

1. A compound of the formula 1 cm H2H N 5 ED 0 -, CH=CH- C- N =- 0 CoGe wherein 0 H = 0 is a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof. 25 2. A compound of Claim 1 wherein (D - N = 0 XXII is selected from z R13 0 H R16 N-R14 J-06 TH R R17 R17 1 01 0 N 40 _N -N-(CH2)n and N 0 R18 A m - O's I. R21 0 R1 8 R20 45' wherein R13, R 14 and W-' are the same or different and are (lower)alkyi, (lower)aIkenyl, amino(low er)aikyl with the provision that the amino may not be on an acarbon, or hydroxy(lower)alkyl with the provision that the hydroxy group may not be on an a-carbon; R16 is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkyithio, amino, (lower)alkylamino, diflower)- 50 alkylamino, formylamino, (lower)aikanoylamino, carboxy, hydroxy, carboxyl(iower)aikyi, carboxy (lower)aikylthio, hydroxy(lower)aikyl, halo(loweralky], amino(lower)aikyi, (lower)alkoxy(lower)alky].

carbamoyl or N-flower)alkylcarbamoyl, or R16 may represent a divalent alkylene group having 3 to carbon atoms; R17 is (lower)alkyl, (lower)alkoxy(lower)alkyl, halo(lower)alkyi, ally], hydroxy(lower)aikyl with the 55 provision that the hydroxy group is not on the a-carbon, amino(lower)alkyl with the provision that the amino group is not on the a-carbon, or phenyl (lower)aikyi; R18 is hydrogen, (lower)alkyl, (lower)aikoxy, (lower) alkoxy(lower) alkyl, (lower) alkylthio, amino, (lower)aikylamino, diflower)alkylamino, carboxy, hydroxy, carboxy(lower)aikyl. hydroxy(lower)aikyi, amino (lower) alky], formylamino, (lower)aikanoylamino, carbamoyl or W(lower)alkylcarb amoy]; 60 n is an integer of from 1 to 3, inclusive; Z is CH, or, when n is 2, Z also may be S, 0 or N-R19, in which R19 is hydrogen or (lower)alkyl; and R20 and R21 are the same or different and are hydrogen, (lower)aikyi, (lower)alkoxy, (lower)alkyl thio, amino, (lower)alkylamino, di(lower)alkylamino, carboxy, hydroxy, hydroxy(lower)alkyl, amino- 65 67 GB2194790A 67 (lower)alkyi, (lower)alkoxy(iower)aikyl, carboxy(lower)aikyi, carboxy(lower)alkylamino, (lower)alkanoylamino, carboxy(lower)alkanoylamino, carbamoyl or W(lower)alkylcarbarnoyl; or a salt, ester, solvate or hydrate thereof.

3.

4.

5.

6.

7.

8.

9.

11 12 13 14 16 17 18 19 compound of Claim 1 wherein compound of Claim 1 wherein compound of Claim 1 wherein compound of Claim 1 wherein compound of Claim 1 compound of Claim 1 compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 A compound of Claim 1 N=Q is 1-methylpyrrolidinio. 0) N=-Q is pyridinio. a N=-Q is 2-amino-5- thiazolo[4,5-clpyridinio. E) N=-Q is trimethylammonio. 0 wherein N=-Q is 3-aminopyridinio. G) wherein N=-Q is 3-formylaminopyridinio. (9 wherein N=-Q is 3-carbamoylpyridinio. wherein N=-Q is 4-carbamoylpyridinio.

E) wherein N-=Q is 3-aminomethylpyridinio. (D wherein N-Q is 2methylthiazolio. a wherein N=-Q is 3-hydroxymethylpyridinio.

wherein N-=Q is 4-hydroxymethyl-pyridinio. (D wherein N=-Q is 4-(Nmethylcarbamoyl)pyridinio. (D wherein N-Q is 4-carboxypyridinio.

wherein N=-Q is 2,3-propylenepyridinio. E) wherein N-Q is 3carboxymethylpyridinio. (D wherein N=-Q is 4-carboxymethylthiopyridinio. A process for the preparation of compounds of the formula XI H2NT--rs) N (D - 0 CO Oe CH=CH- c- N = 0 E) wherein---Qis a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof, which comprises reacting a compound of formula 1 with a compound of formula 11 N2N 5 0 EH2E E 00 EW PM2 to give a compound of formula Ill, xl 1 5Q 11 60 68 GB2194790A 68 CH= NI H CH2 CH(PM2 then reacting a compound of formula III With sodium iodide or potassium iodide to give a 10 compound of formula IV, C H = N S 0 H CH2Ct COOCH(PM2 Ill 5 IV 15 2 then reacting compound of formula W with triphenylphosphine or reacting compound of formula 20 Ill with triphenylphosphine to give compound of formula V, 5.0 r",CH=N y (D TQC%(M3 0 COOM W2 then reacting compound of formula V with a base to give compound of formula VI, E H \==77 C H P ( P h) OPN COO CH (PM2 V 25 V1 35 further reacting compound of formula VI with CICH,CHo to give compound of formula VII, 40 S (:y C H =1 L--Nf I', ', CH:CHCH[ COO [H (Ph)2 VII 45 then reacting compound of formula VII with sodium iodide or potassium iodide to give com- 50 pound of formula Vill, j -7,CH = N S 5 \===/ H 0 IN-CH - CH21 ii, COOCHIPM2 Vill 9 69 GB2194790A 69 R then further reacting compound of formula Vill, with a secondary amine HN wherein R, R R' are each methyl or together form pyrrolidine to give a compound of the formula IX, OCHA N S R /Y CH=CHCH2-N-' R' COO CH (P h)2 IX then reacting compound IX with WY, wherein W' is a tertiaryamine QN and Y is chloro, bromo or iodo, to give a compound of the formula X, r-CH=N S e N Y 0 N-13 CH=CHCH2 COOCH(PM2 X or reacting compound Vill with a tertiary amine Q-N, wherein Q is as defined above to give compound X, then treating compound X with Girard Reagent T or HCI to give compound X1, and if desired converting the free acid to a salt, ester, solvate or hydrate thereof.

21. A process as claimed in claim 20 substantially as described in respect of any of the 30 foregoing Examples.

22. A compound as claimed in claim 1 prepared by a process as claimed in claim 20 or 21.

Published 1988 at The Patent Office, State House, 66/71 High Holborn, London WC 1 R 4TP. Further copies may be obtained from The Patent Office, Sales Branch, St Mary Cray, Orpington, Kent BR5 3RD. Printed by Burgess & Son (Abingdon) Ltd. Con. 1/87.