AU610278B2 - Novel intermediates of cephalosporin derivatives - Google Patents

Description

II any Note: No legalization or other witness required D wV en Q fit 250U41/i PHILLIPS ORMONDE AND FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne, Australia P17/2/83 I I I L~ 1

AUSTRALIA

Patents Act

COMPLETE

[ON

78 Int. Class Class Application Number: Lodged: Complete Specification Lodged: Accepted: I ublished: Priority Related Art: APPLICANT'S REF.: Div. of Name(s) of Applicant(s): -BRISTOL-MYERS COMPANY Addresses) of Applicant(s): 345 Park Avenue New York, New York 10154 UNITED STATES OF AMERICA Actual Inventor(s): Harluhiro Yamashita Masahisa Oka Jun Okumura Takayuki Naito fc~yl i ;a L r, t'~ Address for Service is: PHILL IPS, ORMONDE AND FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne, Australia, 3000 Complete Specification for the invention entitled: NOVEL INTERMEDIATES OF CEPHALOSPORIN DERIVATIVES The following statenment is a full description of this invention, including the best method of performing it known to applicant(s): P9I 1 84 Note: No legalization or other witness required Assistant Genra CZ' To: The Commissioner of Patents P18/7/78 2SoIca S PHILLIPS ORMONDE k FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne, Australia L' F This application is divided from Australian Patent Application No. 40862/85 in which is described and claimed 3-[3-(quaternaryammonio)-l-propen-l-yl]-3-cephem derivatives and processes for the preparation of such derivatives. The present invention is directed to novel intermediates which may be used in the preparation of 3 3 -(quaternaryammonio)-l-propen-l-yl]-3-cephem derivatives and the disclosure of Application No.

40862/85 is hereby incorporated by reference into the present application.

Summary of the Invention This application relates to novel intermediates of the formulae:- (a) N C-r CONH

N

2 H2N S OR 0 CH=CHCH 2

Z

COOH

XXVIII

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 4 I 3

-C-CH=CH-R

3

R

5

R

R

4

-C-CC-R

3

R

COOH

x

R

4

-C-COOH

R

-2- .i i~awar in which R; is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)alkoxy, and R 4 and R 5 are each independently hydrogen, methyl or ethyl, or R' and

R

5 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 carboxyl-protecting groups.

(b)

R

2 3

R

2 4

CH=N

R

2 5

CH=CHCH

2

Z

S0

COOR

2 2

XXIX

wherein R 2 2 is hydrogen or a conventional carboxyl-protecting group, and R 2 3

R

24 and R 25 are the same or different and are hydrogen, hydroxy, (lower)alkyl or (lower)alkoxy or a salt, solvate, hydrate or ester thereof.

(c) o o CH=CH-CEH 2

N-Q

0 coo XXII *2al~ix~ wherein -GNEEQ is a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof.

This application also relates to processes for the preparation of such novel intermediates.

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 oat ooo 0 0000 o wherein R l is amino or protected amino; R 2 is hydrogen, acyl, optionally substituted aryl, substituted alkyl, alkenyl, alkynyl, 0 .i 0 0: o oi 00 0 0 0 0 o o 00 0 0 0 0 0 0 -2b- _1 3 optionally substituted cycloalkyl, cycloalkenyl or an 0- or S-containing 5-membered heterocyclic ring substituted with oxo group(s); R 3 is hydrogen or alkyl; R 4 is hydrogen, acyloxyalkyl, acylthioalkyl, optionally substituted pyridinioalkyl, optionally substituted heterocyclylthioalkyl, alkyl, halogen, hydroxy or optionally substituted thiazolioalkyl; and R 5 is carboxy or protected carboxy; provided that R 5 is COO when R 4 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, 1983), 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.

B) European Patent Application No. 62,321, published October 13, 1982, discloses cephem compounds of the formula I 2-N \(D wherein R 1 is amino or protected amino; R 2 is an optionally substituted lower aliphatic hydrocarbon group, or cycloalkenyl; and the group of the formula

-N

4 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 R C-CONH- T6 4 CH 2-N

R

wherein R1and R 2are as defined above, R 4is a protected carboxyl group and X- is an acid residue.

C) European Patent Application No. 74,653, published M1arch 23, 1983, discloses cephem compounds of the formula 1C-CONH--- S 2 N COOS CH 2 4 wherein R1is amino or protected amino; Ris an optionally substituted lower aliphatic hydrocarbon group, cyclo(lower)alkyl or cyclo(lower)alkenyl; R 3is (lower)alkylamino, N-protected(lower)alkylamino, di (lower) alkylamino, sulf o(lower) alkylamino, hydroxy(.lower)alkylamino, N-protected hydroxy(lower)alkylamino, acyloxy(lower)alkyl, (lower) alkoxy (lower) alkoxy (lower ),'lkyl, di (lower) alkylamino- (lower)alkyl, (lower)alkylthio(lower)alkylt (lower)alkylthio, (lower)alkoxy, (lower) alkoxy (lower) alkoxy, hydroxy (lower )alkoxy, acyl (lower) alkyl, hydr oxy (lower) alkylthio, di (lower) alkylamino- (lower)alkylthio, N-containing unsaturated heterocyclic group, N-containing unsaturated heterocyclicthio, or N-containing unsaturated 5 or 6-rnembered heterocyclicthio (lower) alkyl which may be substituted with suitable substituent(s); and R 4is hydrogen or (lower)alkyl; or a salt thereof.

D) U.S. Patent 4,332,800, issL~ed June 1, 1982 to Tsutomu Teraji et al., discloses inter alia compounds of the formula R 1 -CONH wherein R 1is amino or protected amino; R 2is (lower)alkyl and X is hydrogen or carbamoyl.

E) European Patent Application No. 47,977, published March 24, 1982, discloses cephem compounds of the formula (0)

S

Am- T-C CONH

N

-R 2 0CH R 1 e coo 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); R 2 is hydrogen, optionally substituted alkyl, cycloalkyl or optionally substituted carbamoyl; and R is optionally substituted thiazolio, optionally substituted pyrazolio, tri(lower)alkylammonio or a pyridinio group of the formula Ra in which Ra is substituted (lower)alkyl [the substituent being cycloalkyl, phenyl, hydroxy, alkoxy, halogen, cyano, carbamoyl, carboxyl or sulfo], (lower)alkenyl or carboxy-substituted (lower)alkenyl, (lower)alkylthio or carboxy-substituted (lower)alkylthio, amino or monosubstituted amino [the substituent being (lower)alkyl, (lower)alkanoyl or aminobenzenesulfonyl], di(lower)alkylamino, substituted carbamoyl [the substituent being (lower)alkyl, hydroxy(lower)alkyl, (lower)alkoxy, hydroxy or cyano], di(lower)alkylcarbamoyl, thiocarbamoyl, cycloalkyl, phenyl, hydroxy, (lower)alkoxy, halogen, (lower)alkoxycarbonyl, (lower)alkanoyloxy, (lower)alkanoyl, carboxyl, sulfo, cyano, nitro or hydroxysulfo- (lower)alkyl; Rb is hydrogen or carbamoyl, or has the same meaning as Ra; and Rc is hydrogen or has the same meaning as R and salts thereof.

Although not formally related, European Patent Application No. 25,017, published March 11, 1981, has a similar disclosure.

F) European Patent Application No. 30:630, published June 24, 1981, discloses 3-vinyl cephem compounds of the formula i

I

7

S

R A-CONHN CH=CH N CH=CH 0 2

R

2 wherein R 1 is an optionally protected amino-substituted heterocyclic group which may also have halogen, or a group of the formula R3SO 2 HN 3 o 2H2N in which R 3 is (lower)alkyl; R 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 =NsOR 4 wherein R 4 is hydrogen, cyclo(lower)alkenyl, (lower)alkynyl, (lower)alkenyl [optionally substituted by carboxy or protected carboxy], (lower)alkyl [optionally substituted by one or more of carboxy, protected carboxy, amino, protected amino, cyano, phosphono, protected phosphono and a heterocyclic group which itself may be substituted]; and salts thereof.

This application specifically discloses compounds of the formula 8 in which OR 4 is methoxy, carboxymethoxy, tert-butoxycarbonylmethoxy or 1-tert-butoxycarbonylethoxy.

G) U.K. Patent Specification No. 1,399,086 published June 1975, contains a generic disclosure encompassing a vast number of cephalosporins of the formula

R-C-CO-NH

OR

COOH

wherein R is nydrogen or an organic group, Ra is an etherifying monovalent organic group linked to the oxygen through a carbon atom, B is)S or and P is an organic group. In one embodiment, P may be inter alia a vinyl group of the formula

R

3

-CH=C

4 in which R 3 and R 4 independently may be hydtogen, nitrile, (lower)alkoxycarbonyl, or substituted or unsubstituted aliphatic, cycloaliphatic, araliphatic or aromatic. However, the 1,2,4-thiadiazol-3-yl group is not identified as a possible R substituent and there is no disclosure or suggestion that P may be a quaternary ammonio-substituted 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.

9 H) European Patent Application No. 88,385, published September 14, 1983, discloses compounds of the formula

S

1 S R C---CONH

IR

2

R

3

R

4 in which R 1 is (unsubstituted) bhiadiazolyl; R is carboxy- (lower)alkyl or protected carboxy(lower)alkyl; R 3 is hydroqen, halogen or (lower)alkenyl; and R 4 is carboxy or protected carboxy.

Although 1-propenyl is listed as one of the possible meanings of R the application only exemplifies compounds where R 3 is hydrogen, chioro or vinyl.

I) U.S. Patent No. 4,307,233 issued to Daniel Farge et al. on December 22, 1981, discloses inter alia, 3-vinylcephalosporin derivatives of the formula

S

N CONH

H

2

NN

2 OR5 CHCH-N 4

COOH

in which R 5 inter alia may be alkyl, vinyl, cyanomethyl or a protective group such as 2-methoxyprop-2-yl, and R 3 and 14 are alkyl groups (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R 3 and R 4 taken together with the niti'ogen to which they are attached, may form a saturated heterocyclic ring of 5 or 6 members, optionally containing another hetero-atom selected from N, O and S, and optionally substituted by an alkyl group. The compounds are useful as intermediates in the preparation of 3-thiovinyl cephalosporin derivatives. There is no disclosure or suggestion of a I I 1,2,4-thiadiazol-3-yl moiety in place of the 2-aminothiazol-4-yl substituent or of a quaternary ammonio-substituted prop-nyl moiety for the 3-substituent. Published United Kingdom P? Lnt Application No. 2,051,062 is concordant thereto and has a similar disclop.ire, J) European Patent Application No. 53,537, published June 9, 1982, disclos.s, inter alia, 3-vinylcephalosporin derivatives of the formula CONH

S

S c CH=CH-N /R3 2 0a-R b 0 R 4 5

COOR

2 in which R a and R are the same or different and are hydrogen or alkyl, or taken together, form an alkylene group containing 2 or 3 carbon atoms, R is an acid protecting group, R 2 is an acid protecting group such as an ester, R 3 and R 4 are the same or different and are hydrogen, alkyl (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R 3 and R 4 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, O and S, and optionally substituted by an alkyl group.

The compounds are useful as intermediates in the preparation of 3-thiovinyl cephalosporin derivatives. There is no disclosure or suggestion of a 5-amino-l,2,4-thiadiazol-3-yl moiety in place of the 2-aminothiazol-4-yl substituent or nf a quaternary ammoniosubstituted propenyl group for the 3-substituent.

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

x /2 'PIY1~48t- I I P i I c ii ~i 11 K) European Patent Application No. 53,074, published June 2, 1982, generically discloses a vast number of 3-vinylcephalosporin derivatives of the formula R° -NH- Ia

COOR°

2a wherein R la (in one of several embodiments) may be in which R inter alia may be hydrogen, alkyl, vinyl, cyanomethyl, an oxime-proteting group such as trityl, etc., or a group of the f.ormula

-C-COORC

R 5 R in which R 5 and Rb 5 are the same or different, and may be hydrogen, alkyl or, taken together, an alkylene radical of 2 or 3 carbon atoms, and R 5 is hydrogen or an acid-protecting radical; R°2a is hydrogen or an acid-protecting radical such as methoxymethyl; RO (in one of several embodiments) may be a methyl group substituted by a 5- or 6-rembered aromatic heterocyclic ring containing a single hereto atom, such as 2- or 3-pyridyl, 2- or 3-thienyl or 2- or 3-furyl; and carooxyi-proteciLIg Lgruup, diu n- d a u dLr the same or different and are hydrogen, hydroxy, (lover)alkyl or (lower)alkoxy and z is chloro, bromo or iodo; or a salt, solvate, hydrate, or ester thereof.

12

R

3 i group of the formula

R

4

SO

2 0in which R 4 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

R°

-CH=C-SR

which are stated to have antibacterial activity.

Although this patent includes the possibility of Ro being a I methyl group substituted by an N-containing heterocyclic ring, in both the intermediates and final products (thus giving a heterocyclicsubstituted 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 ammonio-substituted propenyl group, The reference exemplifies Ro 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 (-03SR or -SR, respectively). Also there is no disclosure or suggestion of a 5-amino-l,2,4-thiadiazol-3-yl moiety in place of the 2-aminothiazol-4- yl substituent.

SL) European Patent Application No. 53,538, published June 9, 1982, discloses, inter alia, 3-vinylcephalosporn intermediates of the formula NUV~L LIi£1IJi UI' UJhIHALU6J9URiN DERIVATIVES The following statemaent is a full description of this invention, including the best method of performing it known to applicant(s): P19/3/84 (0) n~f

COOH.

in which n is 0 or 1, R5is hydrogen, al~kyl, vinyl, cyanc'methyl or an oximne-protecting group, and RP3 is halogen. There iz no disclosure or sugg~estion of a 5-amino-1,2,4-thiadiazol-3-yl moiety in place of the 2-arinothiazoJ,-4-yl substituent, and no disclosure or suggestion of a 3-halo-l-propen-l-yl substituent in the 3-positioi.

14 Complete Disclosure Australian Patent Application No. 40862/85 relates to novel cephalosporin derivatives which are potent antibacterial agents. More particularly, it relates to compounds of the formula

'S

N CONH

C

NN G RH J S OR2 CH=CH-CH 2

-NEQ

coP wherein R is hydrogen or a conventional amino-protecting group,

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 ators, or a group of the formula

R

4

R

4

COOH

3 3 -C-CH=CH-R -C-CSC-R or 15 R R x 4 -C-COOH 51 in which R 3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)alkoxy, and R and R are each independently S hydrogen, methyl or ethyl, or R and R taken together with the carbon atom to which they are kttached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and

-N--Q

-2a- I I-

I

is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof.

Also included within the scope of the invention are the solvates (including hydrates) of the compounds of Formula I, as well as the tautomeric forms of the compounds of Formula I, e.g. the 2-iminothiazolin-4-yl form of the 2-aminothiazol-4-yl moiety.

n ;i ii As shown in the structural formula, the compounds of Formula I have the "syn" or configuration with respect to the alkoxyimino group. Because the compounds are geometric isomers, some of the "anti" isomer may also be present. Application No.

40862/85 comprises compounds of Formula I containing at least 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 VIII and IX) also form geometric (cis and trans) isomers about the double bond of the propenyl group.

Both the cis and trans 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 asacetic, trifluoroacetic, citric, formic, maleic, oxalic, succinic, benzoic, tartaric, fumaric, t'andelic, 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.

I

-2b- 16 Examples of physiologically hydrolyzable esters of the compounds of Formula I include indanyl, phthalidyl, methoxymethyl, acetoxymethyl, pivaloyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, 5-methyl-2-oxo-l,3-dioxolen-4-ylmethyl and other physiologically hydrolyzable esters known and used in the penicillin and cephalosporin arts. Such esters are prepared by conventional techniques known in the art.

The compounds of Formula I in which R is hydrogen exhibit high antibacterial activity against various Gram-positive and Gram-negative bacteria, and are useful in che 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 and 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 reconsritution 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 Application No.

40862/85may, 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 I. 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 particular nature and severity of the disease, and is within the discretion of the physician. However, the dosaqe 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 ~_11 1 IL I I iLT-- II- i- 17 of the compounds may be desirable in the case of Pseudomonas infections.

The quaternary ammonio group of the formula

-N=Q

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 .R6 S7

-N-R

18

R

S R 6 7 8 in which R R and R 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 hydroxy gr-up may not be on an a-carbon, (lower)alkoxy with the provision that the alkoxy group may not be S 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)alkylamino, 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 alkyl, (lower) alkoxy (lower )alkoxy (lower) clkyl, di (lower) alkylamino- (lower)alkyl, (lower)alkylthio(lower)alkyl, (lower)alkylthio, systems include, for example, those of the formulae

R

10 0

G

-N

-NR

s K R 1

-N

S

G

-N-N9 1 1 0 0" and the like, in which R9 and Ri are the same or different and U- *~1 19 may be, for example, hydrogen, halogen, amino, (lower)alkyl, (lower)alkenyl, (lower)alkylthio, carboxy, hydroxy, (lower)alkoxy, (lower)alk.,xy(lower)alkyl, halo(lower)alkyl, hydroxy- Clower)alkyl, amino(lower)alkyl, Clower)alkylamino(lower)alkyl, di(lower)alkylamino(lower)alkyl, (lower)alkylamino, di(lower)alkylamino, carboxy(lower)alkyl, carboxy(lower)alkylamino, carbo .y(lower)alkylthio, carbamoyl, N-(lower)alkylcarbamoyl, formylaminc, acylamino, acyloxy, phenyl, pyridyl, amidino, guanidino and the like. Where the structure of the heterocyclic ring permits, R 9and R 10, taken-together, may be an alkylene group containing from 3 to 5 carbon atoms, e~g. propylene.

Examples of combined acyclic/cyclic quaternary ammorio groups include, for example, those of the formulae 12f" 12 12 12

-N

11 7 <1 R Il N

H

R

12 G/ 12 st R 12 R2 KR 12 C4) G) 1 'IN N- (lower) alkyl, -N R 1 R11 7 and the like, in which R may be, for example, (lower)alkyl, (lower) alkoxy (lower) alkyl, hydroxy(iower)alkyl with the provision that the hydroxy may not be on an a-carbon, carboxy (lower) alkyl, amino (lower )aJ kyl with the provision that the amino may not be on an a-carbon, (lower)alkenyl, halo (lower) alkyl, allyl and the like, and R may be, for example, hydrogen, hydroxy, halogen, (lower)alkyl, hydroxy (lower) alkyl, (lower) alkoxy (lower )alkyl, halo (lower) alkyl, amino (lower) alkyl, Clower)aikoxy, (lower)alkylthio, (lower )a3.kenyl amino, (lower )alkylamino, di(lower)alkylamino, acylamino, acyloxy, Icarbamoyl, amidino( lower )alkyl, phenyl, pyridyl, amidino, juanidino and the like.

Preferred quaternary-ammonio groups are those of the formulae R 13 ®I 1.4

R

~NS 16 and R 21 13 4 15 wherein R 13

R*

4 and R are the same or different and are (lower)alk yl, (lower)alkenyl, amino(lower)alkyl with the provision that the amino ma.y not he on an a-carbon, or hydroxy- (lower)alkyl with :he provision that the hydroxy group may not. be on an a-carbon; R 6is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkylthio, amino, (lower )alkylamino, di (lower) alkylamino, formylamino, (lower )alkanoylamino; carboxy, hydroxy, carboxy(lower)alkyl, car boxy (lower) alkylthio, hydr oxy (lower) alkyl, halo(lower)- 21 alkyl, arino(lower )alkyl, (lower)alkoxy(lower)alkyl, carbarnoyl or N-(lower)alkylcarbamoyl, or R 16may .represent a divalent alkylene group having 3 to 5 carbon atoms; R 17 is (lower)alkyl, (lower)alkoxy(lower)alkyl, halo- (lower)alkyl, allyl, hydroxy(lower)alkyl with the provision that the hydroxy group is not on the ca-carbon, ami>.o(lower)alkyl with the provision that the amino group is not on the a-carbon, or phenyl (lower )al kyl; R 18 is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkoxy (lower jalkyl, (lower )alkylthio, amino, (lower )alkylamino, di (lower )alkylamino, carboxy, hydroxyj, carboxy (lower) alkyl hydr oxy (lower) alkyl, amino (lower) al kyl, formylamino, (lower)alkanoylamino, carbamoyl or N-(lower)alkylcarbamoyl; n is an integer of from 1 to 3, inclusive;.

Z is CH 2 or, when n is 2, Z also may be S, 0 orc N-R 1 in which R 19 is hydrogen or (lower)alkyl; and R 20and R 21are the same or different and are hydrogen, (Iower)alkyl, (lower)alkoxy, (lower )alkyltniio, am~ino, (lower)alkylamino, di(lower)alkylamino, carboxy, hydroxy, hydroxy- (.lower)alkyl, arino(lower)alkylf (lower)alkoxy(lower)alkyl, carboxy (lower) alkyl, carboxy (lower) alkylainino, (lower )alkanoylamino, carboxy(lower)aJlkanoylamino, carbamoyl or N-(lower)alkylcarbamoyl.

Particularly preferred quaternary ammonio groups are tl-(lower)alkylpyrrolidinio (and especially N-methylpyrrolidinio), tri(lower)alkylammonio (and especially trimethylammonio), pyridinio, aminopyridiniot formylaminopyridinio, carbamoylpyridinio, amino(lower)alkylpyridinio, carbcxypyridinio, hydroxy(lower)alkylpyridinio, N-(lower)alkyltcarbamoylpyridinio, (lower)alkylenepyridinio, 2-methylthiazolio a~d 415-c Jpyridinio.

In the compounds of Formula 1, particularly preferred values of Rare (lower)alkyl (and especially methyl), cycloalkyl containing from 3 to 5 carbon atoms, l-carboxycycloalk-.-yl containing from 3 to 5 carbon atoms, allyl, propargyl and carboxy(lower)alkyl (an' especially 2-carboxyprop-2-yl). The most preferred compounds of Application No. 40862/85 are 1) 7-2 -m n f -h a i z l 3-l -e h x iio c tm d l 3- (tr imethylammon io) propen-1-yl 1-3-cephem-4-ca rboxyl ate, 2) 7-2 -m n -h a i zr-3y )2Te h x iio c tm d l 3-[3-Cl-methylpyrrolidifio)-l-propen-yl-l3cephem4-crboxylate, 3) 7-2(-mn-f,-~aizl3y)2mtoymnaea~dl 3-3prdno1poe--l--ehm4croyae 4) 7-2 -m n -h a i z l 3-l -e h x iio c tm d l 3-3(-mnprdno--rpn--l--ehm4cr~yae 7-2 -m n -h a iz l 3-l -e h x iio c tm d 3-[3-(3-formylaminopyridinio)-l-propen--yl-3-cephe- 4 ca rboxylate, 6) 7-(2.-(5-amino-1,2,4-thiadiazol-3-yl)-2-fethoXyimiloacetarridoV- 3-(3 -mnmtypriiiQ)--roe-1y ehe-4 carboxylate, 7) 7- 2 -m n h a i z l 3-l -e h x iio c tm d l 3-t3-(3-carbamoylpyridifio)-l-propen1yl-l3cephem- 4 ca rboxyl ate, 8) 7-2(-mn-,,-haizl3-l--ehxiioctmdl 3-[3-(4-carbamoylpyridinio)-l-propel-1-yl1-3-cephem-4carboxylate, 9) 7-2(-mn-,,-haizl3y)2mtoymnaeardl 3-3(-ehlhaoi)lpoe--l--ehm4 carboxylate, 7-2(-mn-,,-haizl3-l--ehxiioctt~dl 3-(3-(2-amino-5-thiazolo(4,5-c)pyridifio)l-propen-llV 3 cephem-4-ca rboxyl ate t 1 1) 7-(2-(5-amino-1,2,4-thiadiazol3-yl)2fethoxyiminoacetarnido)- 3-(3 -y xmtyly dno)-Ip pn -lI pe-4 ca rboxyl ate, 12) 72(amino-1,2,4-tiadiazo-3-y)-2-methoxyirminoaceta~UdoVintermediates in the preparation of 3-thiovinyl cephalosporin derivatives. There is no disclosure or suggestion of a 23 3- (3-hydr oxymethylpyr idini o) -l-pr open- 1-yl -3-cephem- 4carboxylate, 13) 7-[2-(5-amino-1,2,4-thiadiazo1-3-yl)-2-methoxyiminoacetamidoJ- 3-[3-(4-(N-methyicarbarnoyl~pyridinio)-1-propen-l-yl]-3cephem-4--carboxylate, 14) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]- 31-[3-(2,3--propylenepyridinio)-l-propen-1--yl)-3-cephem-4carboxylatet 7-f2-(5-amino-l,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetami-do]- 3-(3-(4-carbamoylpyridinio)-2 -propen-l-yl]-3-cephem-4ca rboxyl ate, 16) 7- (5-amino- 1,2, 4-th iadi azol-3-yl) -2-cycl open tyloxyiminoacetamido (4-carbamoylpyridinio propen-1-yl cephern-4-carboxylate, 17) 7-[2-(5-amino-l,2,4-thiadiazol-3-yl)-2-allyloxyiminoacetamido]-3-(3-(4-carbamoylpyridinio)-l-propen-1-yl1-3cephem-4-carboxylatet 18) 7-(2-(5-amino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetamido)-3-[3-(4-carbamoylpyridinio)-l-propen-1-y1]-3ceph em-4-carboxyl ate, 19) 7-(2-(5-amino-1,2,4-thiadi~zol-3-yl)-2-methoxyiminoacetamido)- 3-(3-(4-carboxypyridiv. ,,o)-l-propen-1-y)-3-cephen-4ca rbox<y1ate, 7- amino- 1, 2,f4- hiadiazol- 3-y)-2-et hoxyiminoacetamido I- 3-[3-(4-carboxypyridinio)-2-propen-1-yll-3-cephem-4carboxylate, 21) 7-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2--methoxyiminoacetam-ido)- 3-(3-(3-carbo~ymethylpyridinio)-l-propen-1-yl]-3-cephew-4carboxylate and 22) 7-t2-(5-amino-1,2,4-tbiadiazo2-3-yl)-2-methoxyiminoacetamidoJ- 4-carboxymethylthiopyridinio)-1-propen-1-yl )-3-cephem-4carboxylate.

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

I

24 [Roman Numeral Arabic Numeral Letter (if appropriate) f appropriate

L

The Roman Numeral designates whether the compound is a final product or an intermediate or 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 2 2 substituent R 2 If the particular R 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 R 2 substituent

R

2 when gEierically referring to an R 2 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 -N Q For convenience, the Arabic Numerals and Letters assigned to some of the preferred R 2 groups and quaternary ammonio groups are set forth below.

Arabic Numeral R 2 1 methyl 2 ethyl 3 allyl 4 propargyl a cyclopentyl Letter N Q A 1-methylpyrr olidin io B pyridinio C 2-amino-5-thi azolo[ 4,5-c]pyri din io D tr iethyl anion io E 3- am inopyr Idin io F 3-formylamiriopyridinio G 3-ca rbamoylpyr idin io H 4-c arbamoy lpyr id inio 3- amin amethylpyr idin io J 2-methylthiazolic K 3-hydr oxyrnethylpy ri din io L 4-hydr oxyrnethylpyrid in io 111- 4- (N-methyJlcar bamoyl )pyridinio N 4-c rboxypyr id in i 0 21 23-propyl enepyr idin io P 3-c arboxyme thy lpyrid in io Q 4-ca rboxyme thyl thiopyrid in io In the primary evaluation of the compounds of this invention, the Minimum Inhibitory Concentrations (MIC's) of the compounds were determined by the two-fold serial agar dilution method in Mueller-Hinton agar against 32 strains of test organams in six groups. The geometric means of the MIC's determined in these tests are shown in Table 1.

Table 1 Config. Geometric Mean of MIC (rncg/rnl) at Cmpd. dou~ble (G-)-III No. bond (7) I-lA E/Z=1/1 0.26 0.70 0.05 0.15 10.23 2.4 I-lA E/Z=7/l 0.13 0.35 0.029 0.05 0.17 1.4 I-lB E 0.20 0.40 0.016 0.044 0.11 1.6 I-IB E/Z=1/4 0.35 0.80 0.05 0.11 0.35 I-iC E 0.10 0.20 0.0071 0.033 0.087 3.8 I-lD E/Z=1/1 0.61 1.4 0.10 0.26 0.46 2.4 I-lD E/z=l0/l 0.30 0.53 0.05 0.076 0.26 1.3 I-lE E 0.20 0.40 0.0094 0.029 0.10 1.4 I-lF E 0.15 0.40 0.0094 0.033 0. 099 1.2 1-lG E 0.20 0.35 0.0001.1t 0.033 0.10 1.4 I-lH E 0.20 0.40 0.0l.j 0.043 0.10 0.97 I-11 E 0.80 1 6 0.10 0.20 0.69 3.1 I-1J E 0.17 0.35 0.025 0.076 0.15 1.6 1-1K E 0.35 0.80 0.029 0.044 0.20 I-iL E 0.26 0.61 0.029 0.088 0.15 2.6 1-im E 0.35 0.70 0.029 0.10 0.17 2.3 I-iN E/Z=7/1 1.2 1. 6 0.013 0.066 0.30 5. 7 E 0.17 0.35 0.029 0.033 0.11 14 1I-2H E 0.20 0.40 0.014 0.057 0.15 1.4 I1-2N E 1.2 2.1 0.016 0.11 0.35 4.7 I-2N z 1.4 3.1 0.044 0.15 0 A9 I-3H B 0.23 0.40 0.057 0.10 0.52 1.9 I-4H B 0.26 0.46 0.066 0.11 0.60 2.6 E 0.13 0.40 0,20 0.46 2.1 4.2 I1P E 0.8 1.6 0.013 0.087 0.34 14 I-iQ B 0.7 0.92 0.0095 0.044 0.23 14 z Penicillin-sensitive S. aureus (5 strains) Penicillin-resistantz S. aureus (5 strains) Cephalothin-sensitive E. (2 strains), Ki.

2neumonia (1 strain) and Pr. mirabiis (2 strains) Cephalothin-resistant E. coli (3 strains) and Ki.

pneumoniae (2 strains) M. rorganii (1 strain), Ent. cloacae (2 strains) and Ser. marcescens (2 strains) (G-)-III Ps. aeruginosa (7 strains) Table 2, below, gives the Protective Dose 50 (PD 50 in mice for a number of the compounds of Formula I againist selected microorganisms. Table 3 gives blood levels of various compounds of Formula I upon intramuscular administration of the test compounds to mice at a dosage of 20 mg/kg.

Table 2 PD 0 4 4 Cmpd. No.

I-lB I -lB o I~lC I-lG

I-IJJ

0 1-K 0 1-111 I-iN 0 I-10 0 I-2N S. aureus Smith 0. 44 0. 65 0 .22 0. 96 0.39 0. 35 0. 53 0. 96 2.0 0.26 5.0 E. coli Ju hi 0.028 0.072 0. 013 0.021 0.015 0.029

NT

NT

NT

0.17

NT

P. aeLuginosa A9843A 7.7

NT

NT

5.92 3.9

NT

NT

NT

NT

NT

NT

NT Not Tested Cmpd. No.

I-lB I-1C

I-ID

iI-1H i I -15 I-1K I-1M

I-IN

I-2N I-3H I-4H 28 Table 3

C

max (mcq/ml) 17 21 20 23 19 24 20 24 28 22 19 27 22 '1/2 (min) 21 32 19 16 16 14 23 19 32 20 47 22 32

AUC

(mcg hr/ml) 11 18 11 14 9.7 14 14 18 17 12 16 18 Application No. 40862/85 relates to processes for the preparation of the compounds of Formula I. The preferred procedures are shown below in Reaction Schemes la, Ib and Ic, while an alternative procedure is shown in Reaction Scheme 2.

The abbreviation "Ph" represents the phenyl group. Thus, the -CH(Ph) 2 moiety is the benzhydryl group, which is a preferred carboxyl-protecting group. When R 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.

Reaction Scheme la O N

CH

2 C1 COOCH(Ph) 2 \7 -COOH N2'/O2 HN7OR

III

c

S

N C ONH N 2N COOCH (Ph) 2 Nal or 1(1 N C CONE.

H 2 N N R 2 O00CH (Ph) 2 rv P (Ph)3 C ~4t4 4 0 4*4 4 0 4 4 0 444 4 I/P (Ph) 44 CONH

S

Ne 0 h 2 P (Ph) 3 COOCH (Ph) 2 Ibase N IFCONH H2 r~ S1-1N OR2 ViI CH=P (Ph)3 (Ph) 2 j 1 LiWU JL; I. L I. I= L U Q J. L Y U U 0 C) V J. '3 1,J III HN S OR 2 H2N Ph) 2 Nal or KI 0 0 C -ONH H 2~ OR 2

N

{(secondary amine) N CONH

H

2 NJS" OR E=CBCB 2 Ph) 2 Q _1q i ary amine) COOCH (Ph) 2 JR1"

Y

CONH

.XlI

Y

COOCH (Ph) 2 31 deblock

S

N C CONE

H

2 N S OR 2 =CHC 2 co Reaction Scheme la shows two alternate means of going from Compound IX to Compound XII. The direct route, utilizing a 4; tertiary amine is applicable for the preparation of all S compounds of Formula I. The indirect route, via Compound X, utilizes a secondary amine as reactant, and is quaternized in the following step. The secondary amine RR'NH may be acyclic (e.g.

dimethylamine) or cyclic pyrrolidine), and this indirect procedure therefore is suitable for the preparation of compounds S of Formula I in which the quaternary ammonio group is acyclic or S "mixed" acyclic/cyclic. This indirect route is not suitable for the preparation of compounds of Formula I wherein the quaternary nitrogen is in a fully unsaturated heterocyclic ring (e.g.

pyridinio, thiazolio, 2-amino-5-thiazolo[4,5-c]pyridinio, and the like).

F Reaction Scheme lb

II

V

and the like, in which R' and R~u are the same or different and 0 CH N l COOCH (Ph) 2

XIII

NaI or KI a

CH="

XITv HI2 (Ph) 2 P (Ph )3 I 1 ~4I~ 0*t 9 I P (Ph) 0a IH 2 P (Ph) 3 (Ph) 2 Ibase 0 N CH=P(Ph) 3 COOCH (Ph) 2

XVI

33 /lC 2 CHNO

XVI

NC. CH=CHCH 2 C1 OH (Ph) 2 Girard Reagent T or

S

OT N C CH=CHC

H

2 C1 COOCH (Ph)2 N -COOHII

OR

Vill as in Scheme la' I Reaction Scheme lb is a variation of RAeaction Scheme la in that the 7-amino, group of the starting material (II) is protectei as a Schiff base during jiiost of the reactiot steps and the desired 7-side chain acid is added'later in the synthesis.

otherwise, the general procedure is similar.

alKyl, carboxy(iower)alKyitfllo, nyaroxytiower)alKy, naioti.ower)- 34 Reaction Sch~eme lc T 11CH=CH-CH 2 C1 COOCH (Ph) 2 [NaI or KI XV I S

XIX

COOCH (Ph) 2 44

HN

N

R

(secondary amine) aX "CH=CHC 2

NA

COOCH~~(Ph) 2 R11 Y Q N X I (tertiary amine) yoS

XXI

COOCH (Ph) 2 L4, b-c 1pyrnliLo.

H

2

N

O CH=CHCH 2

-N-Q

XXII O

COO

N-acylation with III N C CONH N N

SOR

2 0

COO

CH=CHCH

2

-N-Q

Reaction Scheme Ic is a further variation of Reaction Scheme lb. In Reaction Schemes la and lb, quaternization of the 3-side chain is the last step, but in Reaction Scheme Ic the last step is acylation of the 7-amino group. The relationship between Reaction Schemes la, lb and Ic is shown in the following flowchart.

12 7- amin o- 1,2, 4 -th iadiaz ol-3 yl)2-me th oxyimi no a e tami do I H 2 N Il 10 N\OR 2 C 2

C'

IV COOCH(Ph) 2

CH

2 Cl P h 2 \c xiic tlb lc

\Y

C-cc -IT- I I H 2 N S.-IN N\ OR 7-N-Acylation lb T _-C=iH2c WVill CX1(Ph) 2 (Ph) 2 VniI Quaternization Deblcocking Quaternization lc I Deblocking n 2NP XXI I .7-N-Acylation lc Compound I *7"~ag3W-M-«.li. I-i-lliII. in.- I 37 In Reaction Schemes la, lb and Ic, the benzhydryl group was shown as the preferred carboxyl-protecting group. It will be appreciated by those skilled in the art that c'her carboxylprotecting groups, well-known in the art, may be used. The acylating acid III may be used in the form of a derivative sucn 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 aminoprotecting groups, e.g. N-trityl, N-formyl, or the like, The base used to convert the phosphonium iodide (VI or XV) to give the phosphorylide (VII or XVI) may be NaOH, Na 2

CO

3 IRA-410 (OH-) resin, IRA(CO 3 resin, or the like, or a mixture thereof. The chloroacetaldehyde used to convert the phosphorylide VII to the 3-chloropropenyl-3-cephem compound VIII (or Compound XVI to Compound XVII) may be the commercially available 40-50% aqueou.

solution, a distilled solution 70%) or the anhydrous aldehyde.

We have found that Compound VIII prepared from Compound VII (Scheme la) typically had a Z:E ratio of about 2:1 at the propenyl double bond, Compound VIII prepared from Compound XVIII (Scheme lb), on the othei 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 VI'I or XVI to XVII). We have also found that the use of an appropriate silyl reagent such as N,O-bis(trimethylsilyl)acetamide in the Wittig reaction (VII to VIII in Scheme la and XVI to XVII in Scheme lb) caused improvement of the yields and purity of VIII and XVII. The reaction is preferably carried out with equivalents of the silyl reagent. When the chloropropenyl cephem (VIII) was reacted with Nal in acetone to give the iodopropenyl cephem the double bond in the propenyl group was isomerized from Z to E during the iodination. A short reaction period retained the configuration of 'the parent Compound VIII to a large extent, while a long reaction period gave primarily the E isomer of Compound IX. However, an excessive reaction time at high L i i..

38 temperature gives a lower purity compound IX. We find that about minutes at 25 0 C and 2 hours at 5 0 C gives pure IX in good yield. When utilizing Reaction Scheme Ic, we have found That, when iodinating compound XIV with Nal, a p:'rer coimpund is obtained if the acetone solution is dilu-ed with Ci 4 when iodination is essentially completed, and the isomerization portion of the reaction is conducted in the acetone-CCl 4 mixture.

When iodination of the chloropropenyl cephem (XVII) to the iodopropenyl cephem (XIX) 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 minutes at room temperature to give pure XIX without dilution with CCl 4 in the course of the reaction.

Compound XII normally was deblocked without purification, and the final product was purified by reverse phase column chromatography utilizing a glass column containing the packing removed from a Waters' Associates PrepPAK-500/C 1 8 cartridge.

Reaction Scheme 2 N 1

OR

s

CONH

N

CH=CH-CH

2 I1 COOCH (Ph) 2

XXIII

N

1 N 2

XXIV

CH=CHi-CH 2I COOCH (Ph) 2 R~HNS N 2 CONH I W9 1 C CHCH-CH 2

N=

COOCH (Ph) 2 IReduction

XXV

N-

OR2 CH=CHi-CfH 2

NEQ

I (Ph) 2

XXVI

nJC deblock C CONH H2N OR 2 CH=CH N=Q cocP Reaction Scheme 2, shown in brief outline form above, is similar to Reaction Scheme la except that Compound XXIII (equivalent to Compound IX of Reaction Scheme la) is converted to its S-oxide prior to quaternization. Compound XXV is subsequently reduced, and the remainder of Reaction Scheme 2 is a Reaction Scheme la. In Reaction Scheme 2, it is preferred to protect the amino group of the 7-side chain with a known amino-protecting S group such as the trityl group.

The acylating acids of Forn.ula III 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 2 of compounds of Formula III wherein R is methyl, ethyl, propyl and isopropyl. U.S. Patent 4,390,534, referred to in the Prior Art section, above, exemplifies the preparation of a wide variety of compounds of Formula III wherein R 2 is, for example, cyclopentyl, 2-cyclopenten-l-yl, allyl, 2-propynyl, 1-tert. butyloxycarbonyl-lmethylethyl, 1-tert. butyloxycarbonyl-l-cyclopentyl, 1-ethoxycarbonyl-l-methylethyl, tert. butyloxycarbonylmethyl, 1-tert.

butyloxycarbonyl-2-methylpropyl, trityl, and the like.

Compound II herein (7-amino-3-ch.loromethyl-3-cephem-4- I- 41 carboxylate), used as a starting material in Reaction Schemes la, lb and Ic, 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.

Application 40862/85 also provides a process for the preparation of compounds of the formula N C C lR S

OR

G

H=CH-CI 2-N=Q cocP o 4 00* 0P 0 4 0 wherein R 1 is hydrogen or a conventional amino-protecting group, R 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 o'e 1 3

-C-CH=CH-R

'5

R

R

4 3 i -C-C=C-R

R

COOH

x R 4

COOH

R

in which R 3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)alkoxy, and R 4 and R 5 are each independently hydrogen, methyl or ethyl, or R 4 and R 5 taken together with the H2N N OR

III

carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and -N Q is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof; which process comprises reacting a compound of the formula 0 N C CONH 2 2S ON HN OR 2 0 CH=CHCH 2

Z

COOB

1 2 wherein R is the same as R or is a group of the formula i i ii U I i i i I R4

C-COOB

1

R

4 5 1 in which X, R 4 and R are as defined above, B is a conventional carboxyl-protecting group B2 is 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 and, if m is 1, reducing the sulfoxide by conventional means, and subsequently removing all blocking groups by conventional means.

Application No. 40862/85 also provides a process for the preparation of compounds of the formula CIC COOCH(Ph)

ICH

2

CHO

N-T C CO 2coG R1lt S OR 2 H=CH-CH2-N-Q wherein R 1 is hydrogen or a conventional amino-protecting group,

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

R

4 3

-C-CH=CH-R

3 15

R

R

4

S

3

-C-C=C-R

R

i 1 i i rri I i I

I

1 i 1 i!

R

4

CCOOH

R

i-n which R 3 is hydrogen, (lower)alkyl or carboxyl, r is halogen, hydroxy or (lower)alkoxy, and R 4 and R 5 are each independently hydrogen, methyl or ethyl, or R 4 and R taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and

-N=Q

is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof, which process comprises acylating a compound of the formula H2

N

XII

0c H=CHCH 2

-NQ(

COOCH(Ph) 2 CH=CHCH 2-N= Q coo e

XXII

with an acid of the formula N C--COOH 2HN OR 2' BHN S OR

III

2 2 or with an acylating derivative of said acid, wherein R is the S same as R or is a group of the formula

S.

04

I~

COOB

1

X

R

4

-C--COOB

1

R

5 1 in which X, R 4 and R 5 are as defined above, B is a conventional carboxyl-protecting group and 92 is hydrogen or a conventional amino-protecting group.

The reactions are carried out in a non-aqueous organic solvent such as dimethyl sulfoxide, hexamethylphosphoramide, methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile and the like, or mixtures of such solvents. The reactions are conveniently carried out at a temperature of from about -10 0 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, XIX, XXIII or XXIV; we normally prefer to utilize from about 25% to 100% excess of the tertiary amine.

Carboxyl-protecting groups suitable for use as B 1 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 diphenylmethyl (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 carboxyl-protecting groups which are readily removed by treatment with acid. Particularly preferred carboxyl-protecting groups are the benzhydryl and t-butyl moieties.

Amino-protecting groups suitable for use as B 2 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 preferred.

When the cephalosporin nucleus is utilized in the form of the 1-oxide (m 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 alkoxysulfonium salt itself is readily prepared by treatment of the 1-oxide with, for example, acetyl chloride.

This invention relates to novel intermediates of the formula %..LAn k" 2

N-

H2N

S

CH=CHCH

2 z

XXVIII

wherein Z is chloro, bromo, or iodo, R 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

R

I 3

-C-CH=CH-R

15

R

R

4 1 3

-C-C=C-R

R

COOH

4 4 4* 4 44 4 4 .44,e

R

4

-C-COOH

R

in which R 3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, hydroxy or (lower)alkoxy, and R 4 and R are each independently hydrogen, methyl or ethyl, or R 4 and R 5 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 carboxyl-protecting groups.

In another aspect, this invention relates to novel intermediates of the formula Otherwise, tne genera.± pLrotUUit Z:DL1A.LL 47 R 2 3 R 2 4

/CH=N

R 2.5

CH=CHCH

2

Z

C00R 22

XXIX

wherein R 2is hydrogen or a conventional carboxyl-protecting group, and R2 R24and R 25are the same or different and are hydrogen, hydroxy, (lower)alkyl or (lower)alkoxy; or a salt, solvate, hydrate or ester thereof.

In still another aspect, this invention relates to novel G intermediates of the formula 0 0 2 o 0 0O CH=CH-CH 2

-N=-Q

wherein is a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof.

As used herein, the terms acylamino and acyJloxy refer to an acylated amino or acylated hydroxy group in which the acyl moiety is (lower)alkanoyl formyl, acetyl, propionyl, butyryl, isobutyryll isovaleryl, etc.), aroyl benzoyl, etc.), (lower)alkanesulfonYl mesyl, ethanesulfonyl, etc.) or arylsulfonyl benzenesulfonylt tosyl,

AA±L

COOCH (Ph)2 48 As usedi herein, the terms (lower) alkyl" "(lower )alkoxy", "(lower )alkylthio" (or the like) mean straight or branched chain alkyl, alkoxy, alkylthio (or the like) groups containing from 1 to 6 carbon ahoms, inclusive. Similarly, the terms (lower)alkenyl and (lower)alkynyl mean alkenyl or alkynyl groups containing from 2 to 6 carbon atoms.

4.44 44 404 4 *0 0 (1 49 Example 1 N CCONH H H=CH-CH 2 2 N S NOCH oc 0*

CE

3 I-1A *Z/E1/1 2- (5-Amino-i, 2, 4-thiadiazol-3-yl )-2-rnethoxyiminoacetanido 13-( 1-methylpyrrolidinio)-l-propen-1-y1 1-3-cephem-4-carboxylate (I-lA) To a solution of diphenylmethyl 7-[2-(5-amino-J.,2,4thi adi azol- 3-yl) )-2-methoxyiminoacetamido 1- 3- 3-i odo-l1-pr open- yl)-3-cephem-4-carboxylate (IX-l) 150 mg, 0.21 mmole) in ethyl acetate (2 ml) was added a solution of 1-methylpyrrolidine (36 mg, 0.42 mmole) in ethyl acetate (I ml) ~.in one portion with stirring. The mixture was stirred for minutes and diluted with isopropyl ether (10 ml) to form a o precipitate, which was collected by filtration. A mixture of the osolid (13 0 mg) f ormi c acid (1 ml) an d concen tr ate d HCl 1 ml) was stirred at room temperature. After 1 hour, the reaction mixture was concentrated under reduced pressure, diluted with water (20 ml) and filtered. The aqueous solution was passed through a reverse phase column (the packing of PrepPABK-500/C 1 8 cartridge, 100 ml),t eluting with water and 10% CH 3 QH. The desired fractions were collected, and concentrated in vacuo to a s~ttall vo~lume and freeze-dried to give 13 mg of the title onpound (I-lA) melting at >280 0 C (dec.).

PXPU*sll~- CBC I i IR VKBr cm- 1 3400, 1760, 1660, 1610.

max UV Phosphate buffer (pH 7) nm (E 1 236 (372), 288 (322).

max 1 cm NMR 6 D 2 0 2.31 Q~ 3.12 (38H, s, N-CH 3 3.6 (58, ppm n, 2-H 3.79 (1H, s, 4.1 (2H, d, J=8, CH 2 4.2 (3H, s, OCH 3 )f 5.36 (18, d, 5.95 (3H, mi, 7-H 3-CH=CH), 6.66 (1/2H, d, J=l0, 3-CH cis), 7.0 (1/28, d, J=16, 3-CH trans).

Example 2

S

N C--C CONH N 2 NCH CH=CHCi 2 H S OCB 3 co I-lB 7-t2-(5-Amino-1,2,4-thiadazol-3-yl)-2-ethoxyiminoacetaiido)-3 [3-pyridinio-l-propen-1-yll-3-cephem-4-carboxylate

(I-IB)

A m .xture of diphenylmethyl 7-[2-(5-amino-112,4thiadiazol-3-y1)-2-methoxyiminoacetamido-3-(3-iodo--propef-lyl)-3-cephem-4-carboxylate (IX-l) (Et 716 mg, 1 mmole), pyridine (158 mg, 2 mioles) in dimethylsulfoxide (DMSO) (1 ml) was stirred for 1 hour at room temperature, To the mixture Was added ethyl acetate (20 ml) to precipitate a solid (620 mg), which was a'lded to formic acid (6 ml) containing sodium bisulfite (60 mg). The mixture was stirred for 30 minutes at 40 0 C and concentrated to dryness. The residue was dissolved in H20 (40 ml) and some insolubles Were removed. The aqueous solution was charged on a column of reverse phase (PrepPAK-500/C 1 8 100 ml) eluting with extent, while a long reaction period gave primarily the E isomer of Compound IX. However, an excessive reaction time at high 51

H

2 0 (300 ml) and 5% aqueous CH 3 OH (800 ml), and the eluate was monitored by uv (254 nm) and HPLC. The fractions aqueous containing the desired product were combined, concentrated to a small volume and lyophilized to yield 40 mg of the title compound melting at >200 0 C (dec.).

IR KBr cm- 1 3350, 1760, 1660, 1600.

max UV Phosphate buffer (pH 7) n 240 (352), 258 (366), max 1 cm 267 (279), 290 (469).

NMR 6

D

2 0+DMSO-d 6 3.74 (2H, br-s, 4.20 (3H, s, sOCH 3 ppm 5.92 (1H, d, J=4.5, 6.15 (1H, m, 3-CH=CH), 7.04 (1H, d, J=16, 3-CH trans), 8.2 (2H, m, Py-H3,5), 8.62 (1H, m, Py-H 4 8.97 (2H, m, Py-H2,U)' Example 3 N C H2N NS OCH 3 I-1B

CH=CH-CH

2 *Z/E=4/1 7- 2-(5-Amino-lf2,4-thiadiazol-3-yl )-2-methoxyiminoacetamido [3-pyridinio-l-propen- 1-yl-3-cephem-4-carboxylate (I-IB) The chloropropenyl compound, diphenylmethyl amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamidoi-3-(3chloro-1-propen-1-yl)-3-cephem-4-carboxylate (VIII-1) 937 mg, mmoles) was added to a stirred solution of pyridine (237 mg, 1 52 3 rmoles) in DMSO (3 ml) 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 ml) to separate the precipitate which was then collected by filtration, washed with ethyl acetate (10 ml) and dried to give 350 mg of the blocked product. The precipitate was treated with formic acid (3.4 ml) containing sodium bisulfite (34 mg) for 30 minutes at 0 C. After removal of the formic acid, the residue was purified by reverse phase column chromatography (packing of PrepPAK-500/C 18 cartridge, 100 ml) by eluting with 5% aqueous CH 3 OH, The fractions containing the des9ired product were combined on the basis of H-PLC analysis, evaporated under reduced pressure and lyophilized to give 41, mg of the title compound (1-13) Mp. >200 0 C (dec.).

IR :v r cm- 1 3300, 1760t 1660, 1600.

UV XPhosphate buffer (pH 7) nm (E 1 237 (386), 250 (377), MCIx 1 cm 258 (369), 265 (347),1 280 (311), N:P. 6D203.45 3,76 (each lii, d, J=16, 2-HI), 4,18 (3Hf s, ppm OCH 3 )f 5.34 (3H, m, CH=-CH-CH 2 5.92 1H, d, J= 4. 5, 7-H),r 6. 58 4/5H t d, J=11,v 3-CE4 cis) 7. 03 d, 3=16, 3-CH trans), 8.12 (24t, M, Py-HU 3 1 5 8.56 m, Py- 4 8.82 (2H, m, Py- 2 1 6 53 Example 4

S

i-C CONH- N UN3

CH=CHCI

2 -N

NN

I-ic 5 -Amin,,-tidiz.-l-2-methoxyirinoaceta)-2mio. 3.

2 -anino-5-thi azolo 4, pyridinio) lprpenlyl>...3cephem 4-carboxylate (IlC- A stirred solution of dipheanylmethyl 7-[2-(5-aino- 1,2, 4-thiadiazol-3-y-2-methoxyiminoacrt i ipdolpropen-1-yl)-3-cephem-4-carbox U,,Iate (IX-l) (E isome 714 mg, 1 mmole), '-aminothiazolo[45-:Jpyridine (prepared a~cording to the procedure of T. Takahashi et al., Pharm. Bull (Japan), 2, 34 (1954)] and dry DMSO (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 sodiun bisulfite (70 mj) were added to the powder (700 mg), and the mixture was stirred for 30 minutes at 40-45 0 c. After evaporation, the residue was triturated with H 2 0 (40 ml). Insolubles were filtered off, and the filtrate was chromatographed over a reverse phase column (PrepPAK-OO/C 18 100 ml), with H 2 0 and

CH

3 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-1c) as a colorless amorphous powder of the E isomer. Yield mg Mp. >200 0 C (dec.), 54 KBr -1 IR V cm 1 3300, 1760, 1660, 1630, 1600.

ma X- Uv XPhosphate buffer (pH 7) nm (E 1% 245 (499), 285 (286).

max 1 cm NMR 6

DMSO-

d 2 +D2 3.86 (38, s, OCH 3 4.98 (1H, d, ppm 5.2 (2H, m, CH=CH-CH 2 5.57 (1H, m, 3-CH=CH)r 5.96 mi, 7.16 (18, d, J=16, 3-CH trans), 8.36 8.45 (each 18, d, J=7, Py-H), 8.92 (18, s, Py-H).

N C0NH-r---r'C I C

H

2 N SN

C=CH-CH

2 -N(C8 3 )3 1400e I-iD *Z/E 1/i 2-(5-Amino-l,2,4 -thiadiazo1-3-yl)-2-methoxyiminoacetanido]-3r inhy a nmmon o -pr open-1-y)- 3-ceph em- 4-ca rboxyl ate (I-lD) To a solution of diphenylmethyl 7-(2-(5-aino-1,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido -3-(3-iodo-l-propen-lyl)-3-cephem-4-carboxylate (IX-l) 490 mi 58 6 mmole) in ethyl acetate (14 ml) was added a 0,1 M trimeth' -i e solution in ether (13.6 ml) in one portion. The inixt,.,e was stirred for 10 minutes and evaporated to dryness, and the residue 4 0 iwas triturated with ether (20 ml). The resulting solid (490 mg) was added to trifluoroacetic acid (0.2 ml) containing one drop of anisole. After 1.5 hours' stirring, the mixture was evaporated to dyness under reduced pressure and the residual oil was triturated with ether (20 ml). The re~u.lting precipitate was collected by filtration and dissolved in 820 (20 ml). Some inzolubles were removed, and the aqueous solution was eluted on a I

C

18 reverse phase column (the packing of PrepPAK-500/C 1 t cartridge, Waters' 30 ml) using watei: as eluant. Fractions containing the desfred compound were combined and conz7ntrated to a small volume and lyophilized to afford 30 mg of the title compound (I-lD) (Z/E 1/1) as a colorless amlorphous powder, melting at >150 0 C (dec.).

KBr IR v cm 1 3300, 1770, 1670, 1605.

max UV XPhosphate h:ffer (PH 7) nm (E 1 236 (389), 287 (343).

max 1 cm NMR 6 D 20 3.45 3.7 d, J=16, 3.81 (lE, S, 2-H), ppm O 4.1 (2H, d, J=8, -CH 2 4.21 (3H, s, OCqH 3 5.39 (1H, d, J=4.5, 5.95 (2H, m, 3-CH=CH 0 000000 6.61 (1/2H, d, J=l1, 3-CH cis), 7.05 (1/2H, d, 0000 J=16, 3-CH trans).

0000 .0 0 000 .0 Example 6 0 0 0 0 0S 0 Q -0 0 N ___CONHC O LT1 "0 2 H 2N SCH=CHCH o/

H

2 NN \S OCH 3 0 2 0 .0 000000 00 0 7- 2-(5-Amino-1,2,4-thiadiazo -1-yl)-2-methoxyiminoacetarr.jdo 3f3-(3-aminopyridinio)-l-propen-1-yli-3-cephem-4-carboxylat e t 0 a_ Diphenylmethyl 7-(2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamdo]-3-( 3-iodo-1-propen-1-yl)-3-cephem-4carboxylate 716 mg, 1 mmole) was added to a stirred solution of 3-aminopyridine (188 mg, 2 mmoles) in DMSO (1 ml).

S56 The mixture was stirred for 1 hour and diluted with ethyl acetate ml). The resulting precipitate was collected by filtration, washed with ethyl acetate and dried to give 520 mg of yellow powder. A nixture of the powder (500 mg), formic acid (5 ml) and sodium bisulfite (50 mg) was stirred for 30 minutes at 40 0 C. The 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/C 1 8 100 ml), with 7.5% aqueous CH 3 0H elution. The fractions containing the desired compound were evaporated and lyophilized to give the title compound (I-lE) (7 mg, melting at >185 0 C (dec.).

IR vKBr cm1 3400, 1765, 1675, 1620, 1600.

max UV Phosphate buffer (pH 7) nm 246 (403), 290 (468.

o0o max Icm 0 0 0000 0 0o NMR 6D20 3.72 (2H, m, 4.14 (3H, s, OCH 3 5.35 (3i, m, 0oo ppm 6-H CH=CH-CH 2 5.9 (1H, C, J=4.5, 6.1 (1H, 00 0'0 0 m, 3-CH=CH), 7.05 (1H, d, J=16, 3-CH, trans), 8.1 o o o o (1H, m, Py-H 5 8.54 (1H, br-s, Py-H 6 8.68 (1H, m, Py-H 4 9.4 (1H, m, Py-H 2 0 0O 0 00 Treatment of IX-1 (716 mg, 1 rmole) with 3-t-butoxy- 0 oo0 carbonylaminopyridine (324 mg, 2 mmoles) by a procedure similar to that described above gave 12 mg of I-1E.

000000 0 4 ooo0 o a0000 0 I .0 57 Example 7 NJ' $-CON N- H2 o OCH H 3

=H

I-KLE *Z/E=1/ 7- (5-Arnino- 2, 4-thiadiazol-3-yl )-2-rnethoxyiminoacetamido -3- 3-amino-1-pyriL~iaio) -l-propen-1-yl J-3-ceph em-4-carboxylate (1-lE) A mixture of diphenylmethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)--2-methoxyiminoacetamido]-3-( 3-i odo-l-pr open-l1yl)-3-cephem.-4-carboxyl ate (IX-l) 500 mg, 0.7 mmole) 0 0 0 0 and 3-aminopyridine (66 mg, 0.7 mxno2.e) in c-imethylsulfoxide (I a0 00 000 0 ml) was stirred for 20 minutes at room temperature. The mixture 0 was diluted with ethyl acetate (10 ml) and ether (10 ml), and the 0 .011 resulting precipitate was collected by filtration, washed with 00 0 ether (10 ml) and dried. The quaternized salt was dissolved in 0 Do formic acid (3 ml) containing concentrated HCl (0.3 ml) and stirred for 1.5 hours at room temperature. The mixture was 000 concentrated to dryness; under reduced pressure. TVhe residtie was 0 00 dissolved in 2% HCl (10) ml) and filtered. The aqueous layer was 00a00 chromatographed on a reverse phase column (PrepPAK-500/C 1 8 100 0 ml). After washing with water (500 ml), the column was elut.ed 000000 o0 with 5% aqueous CHJ3OH The fractions containing the title 0 0 c-oound were combined, concentrated ii vacuo and freeze-dried to 0 give 15 mg of the title compound (I-lE) as a 000000 0 0 colorless amorphous powder. Mp. >160*C (dec.).

0 0 quaternization step, at least one mole of the tertiary amine should be used per mole of Compound IX, XIX, XXIII or XXIV; we 58 I R V r cm- 1 3400, 1765, 1675, 1620, 1600.

max UV X Phosphate buf fer (pH 7) nm (E 1 244 (434), 287 (333).

max 1 cm NMR 6 DMS-d 6 +D2 03. 73 2H, in), 4 .14 (3H, s, 0CH 3 5.35 (3H, PPM in, 6-H CH=CH-CH 2 6.0 (2H, mn, 7-H 3-CH=CH) 6. 6 (1/2H, d, J=11, 3-CH cis) 7. 05 (1/2H, d, J=16, 3-CH trans) 8 .08 (18, mn, Py-H 5 8 .6 (2H, m, Py- 4 6 9.4 (18, in, Py- 2 Example 8

S

N C CONH

HO

H 2 N OCH 3 0C cH CHCH 2 7 2 -(S-Amino-1,2,4-thiadiazol-3-y)--methoxyiminoacetanido]-3- -[3(I-foryaioyiii)lpoe-1y)3cpe---abLl A mixture of diphenylinethyl 7-[2-(5-ainino-1,2,4th dao--l 2mtoymnaetmd (3ioolpoe-1 yl) -3-cephei,,-4-carboxyl ate (IX-1) 716 mng, I mmole) and 3formylaminopyridine (prepared according to the procedure of N.

Enoinoto et al., Bull. Chemn. Soc. Japan, 45, 2665 (1972)] (244 mg, 2 minoles) in DMSO (2 inl~, was stirrod at room temperature for 1 hour, and poured into ethyl acetate (200 ml). The precipitate was collected by filtration, washed well with ethyl acetate and dried. A mixture z f the quaternized salt (500 mg) and sodium bis."'fite (50 mg) in HCQOH (5 ml was s' 1rred at 40-500C for iinues and evaporated to dryness in vacuo, The residue was 59 dissol~ved in water (40 ml), neutralized with NaHCO 3 and then filtered to remove insoluble material. The clear filtrate was chromatographed on a reverse phase column, PrepPAK-500/C 18 (100 ml), with water and 5% CH 3 OH, 10% CH 3 OH, 20% CH 3OH and 30% CH 3 The fractions containing the desired compound were combined, concentrated in vacuo and lyophilized to give 16 mg of the title compound (I-lF) as a tan powder. Mp. >170 0 C (dac.).

KBr I IR :v cm 3340(br), 1760, 1670, 1620(br), 1590.

max Phoshatebuffr (p 7) 1% UV. X Popaebfe p nm (E )218 (428), 248 (362), max 1 cm 290 (474).

NMR 6

D

2 0+a C 3 3.68 (2H, br, 4.15 (3H, s, OCH 3 5.91 PPM (1H, d, J=4.5, 6.25 (1H, m, CH=CH-CH 2 6.98 (1H, d, J=16, 3-CH trans), 8.8-7.9 (4Hjm, Py-H), 9.38 (lH, br, NHCHO).

Example 9

S

N-CONH N CONH2 N 2 H 2 N NOCH 3 0 HC 2 co 1-1G *E 7- 2(5Aminol,2, 4thiadiazol3-yl) 2-methoxyiminoaetamido 1-3- 3- (3-carbamoylpyridinio) -1-propen-.l-yl1]-3-cephem-4-carboxylate (I-lG) To a solution of diphenylmethyl 7-[2-(S-amino-1,2,4thiadiazol-3-yl)-2--methoxyiminoacetamido-3-( 3-iodo-l-propen-1yl) -3-cephem-4-carboxyl ate (IX-1) 716 mg, 1 minole) in DMSO (2 ml) was added nicotinamide (244 mg, 2 mmnoles), and 1 the mixture was stirred at ambient temperature for 1.5 hours and poured into ethyl acetate (200 ml) with stirring. The resulting precipitate was collected by filtration. The quaternized salt (500 mg) was dissolved in HCOOH (5 ml) in the presence of sodium bisulfite mg), and the mixture was heated at 40-50*C for 40 minutes, with stirring, and evaporated to dryness. The residue was dissolved in water (40 ml) and an insoluble solid was filtered off and washed with a small amount of water. The filtrate and wash were combined and chromatographed on a reverse phase column, PrepPAK- 500/C 1 8 (100 ml). After elution with water and 10% and aqueous CH 3 OH, successively, the fractions containing the desired material were combined, concentrated in vacuo and freeze-dried to yield 21 mg of the title compound (I-lG) as a yellow powder. Mp. >175 0 C (dec.).

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

max 0° UV Phosphate buffer (pH 7) nm (E 235 (326), 274 /sh, max 1 cm 405), 290 (446).

o 0 OD O o NMR 6D2 3 3.68 (2H, br, 4.15 (3H, s, OCH3 5.32 Q 0 0 ppm (1H, d, J=4.5, 5.45 (1H, d, J=7, o o o

CH=CH-CH

2 5.88 (1H, d, J=4.5, 6.15 (1H, d-t, J=16 7, 3-CH=CH), 7.00 (1H, d, o o J=16, 3-CH trans), 8.23 (1H, m, Py-H 5 9.03 0 0 0000004 0 00 000 0 C 61 Example

S

N- CONH--- H2 S/ OCH 3 CH=CHCH2 CONH 2 I-1H *E 7 -[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido)-3- [3-(4-carbamoylpyridinio)-l-propen-l-yl)-3-cephem-4-carboxylate (I-1H) To a stirred solution of diphenylmethyl 7-(2-(5-amino- 1,2,4-thiiadiazol-3-yl)-2-methoxyiminoacetamido-3-(3-i.odo-lpropen-l-yl)-3-cephem-4-carboxylate (IX-1) 716 mg, 1 mmole)in dry DMSO (2 ml) was added isonicotinamide (244 mg, 2 mmoles).

The mixture was stirred at room temperature for 1 hour and then poured into ethyl acetate (200 ml). The resulting precipitate was collected by filtration, washed well with ethyl acetate and dried. A stirred mixture of the quaternized material (400 mg) and sodium bisulfite (40 mg) in HCOOH (4 ml) was heated at 40-50 0 C for 1 hour, and evaporated to dryness under reduced pressure. The crude solid war dissolved in water (40 ml). After filtration of an insoluble material, the filtrate was chromatographed on a reverse phase column (packing of PrepPAK/C 1 8 100 ml) using water and 10%, 20% and 30% aqueous CH 3 0H as eluant.

The fractions containing the desired compound were combined, evaporated and lyophilized to give 21 mg of the title compound (I-1H) agi a pale yellow powder. Mp. >180 0 C (dec.).

1 62 K~ 1 IR KBr cm 3340(br), 1760, 1670, 1600.

max UV XPhosphate buffer (pH 7) nm (E 222 (362), 285 (452).

max 1 cm 1MR D 2 O+NaHCO 3 3.68 (2H, br, 4.15 (3H, s, OCH 3 5.33 ppm (1H, d, J=4.5, 5.46 (2H, d, J=7,

CH=CH-CH

2 5.90 (lH, d, J=4.5, 6.17 d-t, J=16 7, 3-CH=CH), 7.02 (1H, d, J=16, 3-CH trans), 8.43 9.09 (each 28, d, J=7, Py-H).

,,,toExample 11 C CONS CH NH 2 N 0N 0 OH.FH CH=CHCH 00 2 N S OCH 3 2 o oE) I-i*E 0 0 o 0 0 0ooo 7 [2-((5-Ami no-1,r2,44-thi adia zol -3-yl 2-metthoxyiminoacetamidol- -3o 0 0 00 o 0 0 (.3-(3-aminomethylpyridino) -l-propen-l-yl -3-cephem-4-carboxylate 0 A mixture of diphenylmethyl 7-f2-(5-amino-1,2,4- 000000 thiadiazol 3 yl)-2-methoxyiminoacetamido]-3-(3-iodo-l-propen-lyl)-3-cephem-4-carboxylate (IX-1) 716 mg, 1 mmole) and 3-(tbutyloxycarbonylaminomethyl)pyridine (516 mg, 2 mmoles) in DMSO (2 ml) was stirred at ambient temperature for 30 minutes. The mixture was poured into ethyl acetate (200 ml), and the precipitate was collected by filtration, washed well with ethyl acetate and dried. A mixture of the quaternized salt (500 mg), sodium bisulfite (50 mg) in HCOQH (5 nl) was dtirred at 40-50 0 C for minutes and evaporated to dryness under reduced pressure. The residual solid was dissolved in water (40 ml), and the mixture 63 was neutralized with NaHCO Insoluble material filtered off, and the filtrate was chromatographed on a reverse phase column (packing of PrepPAK-500/C 18 cartridge, 100 ml), eluting with water, 10%, 20% and 30% aqueous CH 3 OH, successively.

The fractions containing the desired compound were combined, evaporated and lyophilized to provide 10 mg of the title compound (I-lI) as a tan powder.

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

max UV Phosphate buffer (pH 7) nm (E 1 235 (sh, 260), 286 max 1 cm (370).

0 0

I

0100 0 0 0 00 o 00 0P00000 0 0 0 0 SMR 6 D 2O+NaHCO3 ppm 3.68 (2H, br, 4.16 (3H, s, QCH 3 6.98 (lI, d, J=16. 3-CH trans), 8.05 (1H, m,

PY-H

5 8.50 (1H, m, Py-H 4 8,80 (2H, m, Py-H ,6)i Example 12

H

2 N CH=CHCH 2 U-CONH 2 aoas a I-lu 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3t3-(4-carbamoylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-1H) A mixture of diphenylmethyl 7-f2-(5-amino-l,2,4thiadiazo1-3-y2)-2-methoxyiminoacetamidoJ-3-(3-iodo-l-pren-lyl)--cephem-4-carboxylate (IX-l) (E isomar, 4.1 go 5.7 mmoles) and isonicotinamide (1.4 g, 11 mmoles) in dry DMSO (6 ml) was 64 stirred for 2 hours at room temperature while monitoring by TLC (silica gel plate, CHC13:CH 3 OH The reaction mixture was diluted with ethyl acetate (100 ml) to separate a yellow gum, which was treated with formic acid (40 ml) and sodium bisulfite (390 mg) at 45 0 C for 30 minutes. The resulting solution was concentrated to dryness. The residue was dissolved in H20 (100 ml) and insolubles were removed by filtration. The combined filtrate and water wash was applied to the top of a column containing reverse phase packing (PrepPAK-500/C, 8 120 ml). The column was eluted with H20. The eluate was collected in 300 ml fractions and monitored by uv (254 nm) and HPLC (Lichrosorb RP-18, 4 x 300 mm, 0.01 M ammonium phosphate buffer, pH 7.2 cortaining 20% CH 3 OH). Fraction Nos. 4 and 5 were combined and concentrated to a small volume. Lyophilization gave 250 mg of the title compound I-1H, melting at >180°C (dec.).

The spectra indicated that the product was identical to that obtained in Example Preparation of the hydrochloride To a suspension of Compound I-1H (98 mg, 0.18 mmole) in CH3OH (1 ml) was added HC1 (0.1 ml), and the mixture was stirred for 5 minutes. To the resulting ye '.low solution was added acetone (100 ml) to give a precipitate, which was collected by filtration, washed with acetone (2 x 10 ml) and dried in vacuo to give the hydrochloride salt of XI-1H as a colorless powder. Yield 88 mg Mp.

>190C (dec.).

IR V KBr cm- 3300, 1770, 1680, 1620.

max UV XPhosphate buffer (pH 7) nm (E 1% 227 (385), 286 max 1 cm (374).

N MR 6 2 02.32 (1H, acetone-H), 3.79 (2H, br-s, 4.17 PPM IH, s, OC 3 5.34 (1H, d, J=4.5, 5.49 d, J=7, CH=CH-CI-1 2 5.93 (1H, d, J=4.5, 6.28 (1H, d-t J=16 7, 3-CH=CHi) 7 .15 1H, d, J=16, 3-CH), 8.43 9.1 (each 2H, d, J=7, Py-H).

Exampl1e 13

S

N c CONH H N *C

H

2 3#

CH=CCH

COOE

C

I-J.J*E

7 2 -(5-Amino-1,2,4-thiadiazo-3-.yJi-2-methoxYiminoacetamido.3- 3 -(2-methylthiazolio)-l-propen-1-yl-3-cephem4-carboxylate To a mixture of diphenylmethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamidol-3-(3-iodo -l-..propenlyl)-3-cephem-4-carboxylate (IX-l) 714 mg, 1 mmole) and 2methyithiazole (prepared according to the procedure of R. P.

Kurkjy, E. V. Brown, J. Am. Chem. Soc., 74, 5778 (1952)) (198 mg, 2 mm oles) in dry CH 2 Cl 2 (10 ml) was added AgBF 4 (90% pure, 217 mg, 1 mmole) at -20 0 C. The mixture was stirred for 30 minutes at room temperature and filt".ered. The precipitate was extracted with 10% CH 3 QOH-CHC1 3 (3 x 20 ml). The combined extracts were washed with brine (2 x 5 ml), dried over MgSO 4 and evaporated .o dryness to give a yellow residue, which was triturated with isopropyl ether and filtered to yield 350 mg of the quaternized

__A

66 product. A mixture of this solid, sodium bisulfite (35 mg) and formic acid (3.5 ml) was stirred at 40 0 C for 30 minutes. The mixture was concentrated to remove the formic acid, and the residue was diluted with H 2 0 (40 ml). Some insolubles were removed by filtration. The filtrate was placed on a reverse phase column (PrepPAK-500/C, 8 100 ml). The column was eluted with H 2 0 (200 ml), 5% aqueous CH 3 OH (400 ml) and 10% aqueous

CH

3 OH (300 ml), successively. The fractions containing the desired product were pooled on the basis of HPLC analysis (Lichrosorb RP-18. 4 x 300 mm, 0.01 M ammonium phosphate buffer pH 7.2, containing 20% CH 3 OH). The combined solution was concentrated to a small volume and lyophilized to give 40 mg of the title compound (I-1J) Mp. >195 0 C KBr -1 IR v cm 1 3300, 1760, 1660, 1600.

max V Phosphate buffer (pH 7) nm 238 (442), 292 (421).

n;ax 1 cm NMR 6D 2 0+DMSO-d 6 ppm 3.06 (3H, s, thiazole-CH 3 3.74 (2H, br-s, 4.19 (3H, s, OCH 3 5.92 (1H, d, 6.1 (1H, m, 3-CH=CH), 6.8 (1H, d, J=16, 3-CH trans), 8.04 8.23 (each 1H, d, J=4, thiazo.e-H).

67 Example 14 N CONH S H 2 M- sH Ii Co.

H

2 N OCH N CHC 2

H

2

OH

COO 2 I-IL *E 7-[2-(5-Amino-, 2,4-thiiadiazol-3-yl)-2-methoxyiminoacetamido]-3- [3-(4-hydroxymethylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-lL) A mixture of diphenyl 7-[2-(5-amino-l1,2,4-thiadiazol- 3-yl)-2-methcxyiminoacetamido]-3-(3-iodo-l-propen-l-yl)-3-cephem- 4-carboxylate (IX-1) (E isomer,l.07 g, 1.5 mmole), 4-hydroxymethylpyridine (818 mg, 7.5 mmole) in CH3CN (4.5 ml) and CH 3 OH (3 ml) was stirred at room temperature under N 2 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 (3 1, ml) to give 1.28 g of the quaternized cephem ester as a yellow powder. A solution of the quaternized ester (1.25 g) and sodium bisulfite (600 mg) in 85 HCOOH (10 ml) was stirred at room temperature under N 2 atmosphere for one hour. After the addition of 85 HCOOH (5 ml), 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 g of the crude formate of the title compound. A suspension of this compound (1.15 g) in water (100 ml) was filtered to remove insolubles, which were washed with water (10 ml x The filtrate and the washes were combined and subjected to reverse phase column chromatography. The column, which was packed with the packing taken out of a prepPAK-500/C 1 1 3 cartridge column 68 (Waters) 60 ml), 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 (I-lL)as a pale yellow powder. To a suspension of the powder (90 mg) in methanol (9 ml) was added 1 M HC1 in CH3OH ml) and the mixture was stirred at room temperature and concentrated in vacuo. To the concentrate was added isopropanol to precipitate 77 mg of the hydrochloride of the title con~TDound.

Pale yellow powder. M. p. >190 0 C(dec.).

KBr -1 IR v cm 1775, 1670, 1635, 1530.

max Phosphate buffer (pH7) UV max nm 230 (22600), 264 (sh, 16300) NMR 6 D2 ppm 3.83 (2H, br. 2-CH), 4.17 (3H, s, OCH3), 5.06 (2H, s, NV \-CH20H), 5.36 (1H, d, Hz, 5.41 (2H, d, J=7 Hz, CH=CH-CH 2 5.94 (1H, d, J=4.5 Hz, 6.36 (1H, d-t, J=16 and 7 Hz, CH=CHCH 2 7.13 (1H, d, J=16 Hz,

CH=CH-CH

2 8.08 and 8.83 (each 2H, d, J=7 Hz, Py-H).

69 Example C CONH N N

H

2 N S OC2H CH CHCH -N )ONH 2 I-2H *E 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3- [3-(4-carbamoylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-2H) To a solution of 200 mg of 7-amino-3-[3-(4-carbamoylpyridinio)- l-propen-1-ylJ-3,cephem-4-carboxylate hydrochloride (E isomer) in 5 ml of 50 aqueous acetone was added portionwise 190 mg of 2-ethoxyimino-2-(5-amino-l,2,4-thiadiazol-3-yl)acetyl chloride hydrochloride [prepared according to the procedure described in published Japan patent application (Kokai) 57-24389 and the mixture was adjusted to pH 6.5-7.0 with 2 N Na2CO3 (about 1 ml). The reaction mixture was stirred at 10 °C for an hour, acidified to pH 2 with 1 N HCI and evaporated in vacuo. The residue was filtered and the filtrate was chromatographed on a column of HP-20, which was eluted with 500 ml 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 ml) to give 263 mg (93 of the title compound M.

p. 170 OC (dec.).

To a stirred suspension of 225 mg (0.40 mmole) of the above zwitterion in 10 ml of methanol was added 1 ml of 1 N HCI in CH 3 OH and the mixture was stirred at room temperature for minutes. The solution was filtered and concentrated under reduced pressure. To the residue was added 15 ml 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 (57 M. p. 160 OC (dec.).

Estimated purity 65 KBr -1 IR V cm 3300, 1780, 1680, 1620.

max UV Phosphate buffer (pH7) nm 227 max (22300), 288 (22800).

NMR D20 ppm 1.44 (3H, t, J=7 Hz, OCH 2

-CH

3 3.74 (2H, br. s, 2-H) 4.45 (2H, q, J=7 Hz,

OCH

2

-CH

3 5.36 (1H, d, J=4.5 Hz, 5.46 (2H, d, J=7 Hz, 3-CH=CH-CH 2 ),5.92 (1H, d, Hz, 6.20 (1H, m, 3-CH=CH), 7.04 d, J=16 Hz, 3-CH=CH), 8.43 <2H, d, J=7 Hz, Py-HA), 9.10 (2H, d, J=7 Hz, Py-H Example 16 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3- [3-(4-carbamoylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-lH) (E isomer) This Example shows the preparation of Compound I-1H via the last few steps of Reaction Scheme la or Ib, wherein the intermediate benzhydryl 7-[2-(5-amino-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3-[3-(4-carbamoylpyridinio)-l-propen-lyl]-3-cephem-4-carboxylate formate (XXVII-1H) is isolated.

71 A. Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-(4-carbamoyl-l-pyridinio)-l-propen-1-yl]-3cephem-4-carboxylate Formate (E isomer) (XXVII-1H) A solution of XII-lH (Y 1, E isomer) (34 g, pure) in a mixture of acetone and CH30H 200 ml) was placed on a column of Amberlite IRA-410 (formate form 340 ml). The column was eluted with the same solvent system. The first fraction (1 L) was evaporated to about 100 ml of the volume and the brown residue was triturated with isopropyl ether (400 ml).

The resulting powder was collected by filtration and dried under vacuum to afford 29 g (75% pure by HPLC) of the title compound XXVII-lH (E isomer) as a brown powder melting at >150 0 C (dec.).

IR vKBr cm- 3300, 1780, 1680, 1630, 1600.

max EtOH 1% UV XEtH nm (E 282 (186).

max 1 cm acetone-d /CH 08d(1/11 toR 6 O6 3-d/ 4 4.0 (38, s, OCH), 5.26, (18, d, ppm J=4.5 Hz, 5.43 (2H, d, 3=7 Hz, CH 2 N 5.99 (18, d, J=4.5 Hz, 6.5 (18, m, 3-CH=CH), 6.92 (1H, s, CHPh 2 7.1 (1H, d, 2 J=16 Hz, 3-CH), 7.35 (10H, m, Ph-H), 8.36 (18, s, HCOO), 8.46 9.12 (2H each, d, J=8 Hz, Py-H).

B. 7- 2-(5-Amino-1,2,4-thiadiazol-3-yl )-2-methoxyiminoacetamido]-3-[3-(4-carbamoyl-1-pyridinio)-l-propen-1-yl)-3-cephem-4carboxylate (I-1H) (E isomer) A mixture of XXVII-lH (E isomer) from Step A (29 g, pure) and 85% formic acid (290 ml) was stirred for 2 hours at room temperature. Evaporation of the mixture gave a brown oil which was triturated witn acetone (500 ml). The powder was collected by filtration, washed with acetone (2 x 100 ml) and 72 dried in vacuo to give 24 g (50% pure by HPLC) of the crude title compound. The brown solid was treated twice with 2 N HC1 (1 L and 0.5 The aqueous extracts were combined and placed on a column packed with Diaion HP-20 (1.5 The column was washed with water (8 L) and eluted with 30% CH3OH (5 The fraction containing the desired product was evaporated to about 30 ml.

The concentrate was treated with acetone (200 ml) 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 CH3OH (100 ml) was added N HCI in CH 3 OH (55 ml) at room temperature and the mixture was stirred for 30 minutes. The resulting clear solution was filtered to remove insolubles, concentrated to about ml of the volume and precipitated with isopropanol (200 ml).

The resulting powder was collected, washed with isopropanol ml) and dried in vacuo to give 10.5 g (85% pure) of the title compound I-lH (E isomer) (HC1 salt), melting at >180°C (dec.).

Pale yellow powder.

Example 17 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3- [3-(4-carbamoylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-1H) (E isomer) This example shows the preparation of Compound I-1H via the last few steps of Reaction Scheme la or lb, wherein intermediate XXVII-lH (the formate) is not isolated.

A solution of IX-1 (E isomer) (27.6 g, 38.5 mmole) and isonicotinamide (22.8 g, 187 mmole) in a mixture of CH 3 CN (120 ml) and CH,OH (100 ml) was stirred at room temperature for 1 hour -j under a nitrogen atmosphere. After evaporation of the organic solvents, the oily residue was triturated with isopropyl ether to give 50.5 g of a mixture of the quaternized salt and isonicotinamide. A solution of the mixture (50.3 g) and sodium bisulfite (16 g) in 85% HCOOH (160 ml) was stirred at room tempeiature for minutes and subsequently at 40 0 C for 1 hour under N 2 The ;r "j~r?3i~s~- 73 mixture was evaporated in vacuo. The residual oil was mixed with toluene (50 ml), evaporated azeotropically and triturated with acetone (400 ml) to give 27.8 g of the crude title compound.

This material was treated twice with 2 N HC1 (1 L and 0.5 L).

The acid extracts were combined and placed on a column of resin (1.5 The column was eluted with water (9L) and methanol (10L). The fractions containing the desired compound were combined and concentrated to give a yellow oil, which was triturated with acetone (300 ml) to yield 9.35 g of the zwitterion form of the title compound.

To a suspension of the product (9.3 g) in CH 3 0H (180 ml) was added 1 N HC1 in CH 3 OH (55 ml) to obtain a clear solution. The solution was concentrated to about 100 ml and diluted with isopropanol to precipitate 9.50 g (purity 75%) of the title compound I-lH (E isomer) as its hydrochlorid.e. Pale yellow amorphous powder. M.p. >195 0 C (dec.).

Example 18 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3- [3-(4-carbamoylpyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate (I-1H) (E isomer) This example shows the preparation of Compound I-1H via the last step (7-N-acylation) of Reaction Scheme Ic.

To an ice-chilled suspension of the 7-amino-cephem hydrochloride XXII-H (E isomer) (5.0 g, 12.6 mmole) in aqueous acetone (100 ml) 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-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetyl chloride hydrochloride (4.02 g, 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 tlc. After all of Compound XXII-H had been 74 consumed, the mixture was acidified to pH 3 by the addition of 2 N hydrochloric acid. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was diluted with acetone (400 ml) to separate the precipitate, which was collected by filtration to afford 9.59 g 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 ml) was filtered, and the filtrate was adsorbed on a column of HP-20 resin (500 ml). After washing with water the column was eluted with 25% aqueous isopropyl alcohol and the eluate was collected in 100-ml fractions. The desired fractions were pooled, acidified with 2 N hydrochloric acid ml) and concentrated. The residual oil was triturated with isopropyl alcohol (200 ml), and the precipitate was collected by filtration. After drying over phosphorus pentoxide, 5.18 g of the title compound I-lH (E isomer) hydrochloride was obtained as a yellow amorphous powder. M.p. >190°C Estimated purity Examale 19 Purification and crystallization of Compound I-1H (E isomer) Compound I-lH hydrochloride obtained in Example 16 was a pale yellow amorphous powder of 85% purity.

Procedure 1 Six grams of the 85% purity hydrochloride was dissolved in 20 ml of H 2 0 and filtered through a celite pad. The ambercolored filtrate (pH 2) was passed through a reverse phase column (the packing of prepPAK-500/C18 cartridge, Waters; 1,0 ml), which was eluted with water. The eluate was collected in 120-ml fractions with monitoring by HPLC*. Fraction No. 3 through fraction No. 5 were combined and concentrated to about 10 ml, and precipitated by acetone (100 ml) to give 3.3 g of the zwitterion form of I-1H (pale yellow amorphous powder; estimated purity To a suspension of the 95% purity powder (3.2 g) in

CH

3 OH (32 ml) was added N HC1 in CH 3 OH (18 ml), 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 ml. To the concentrate was added isopropanol (100 ml) to separate a pale yellow precipitate, which was collected by filtration, washed with isopropanol (5 ml) and dried to yield 2.6 g of the HC1 salt (amorphous powder; estimated purity A solution of the 95% purity hydrochloride (1 g) in water (4 ml) was adjusted to pH 6.5 with NaHCO 3 (200 mg) and stirred for 30 minutes. The crystals which separated during stirring were collected by filtration, washed with water (2 x ml) and dried in vacuo to give 710 mg of I-1H (zwitterion form) as pale yellow prisms. M.p. >185 0 °C Microanalysis showed it to be the trihydrate.

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

max UV Phosphate buffer (pH7) nm 227 (22000), 290 (23000), max NMR 6 DMSO-d 6

+D

2 0 ppm 3.45 (2H, br, s, 3.9 (3H, s, OCH 3 4.99 (1H, d, J=4.5 Hz, 5.16 (2H, d, J=7 Hz, CH 2 5.61 (1H, d, J=4.5 Hz, 7-H), 5.8 (1H, d-t, J=16 7 Hz, 3-CH=CH), 6.93 (1H, d, J=16 Hz, 3-CH), 8.18 8,89 (each 2H, d, J=7 Hz, Py-H).

Anal. Calc'd for C 2 1H 2 0

N

8

O

6

S

2 3H 2 0: C, 42.14; N, 18.72; Found: C, 42.41; N, 18.86; H, 4.38; S, 10.71.

H, 4.35; S, 11.00.

Column, Lichrosorb RP-18, 4x300 mm: Mobile phase, 0.01 M II_ i 76 phosphate buffer (pH 7.2)/CH3OH 85/15: Detection, uv (254 nm).

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

A solution of the 85% pure hydrochloride (250 mg) in water (1 ml) was treated with charcoal. The solution was adjusted to pH 6.5 with NaHCO 3 (60 mg) and decolorized with charcoal. The 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 (2 x 2 ml) and dried under reduced pressure to give 170 mg (80% recovery) of pale yellow prisms of I-1H (zwitterion form), melting at >185*C which was identical with that obtained by Procedure 1, (as shown by IR, UV,

NMR).

The crystalline zwitterion form of Compound I-1H was slightly soluble in water (6 mg/ml in saline at 23°C).

Example

S

N CONHCH2O H N OCH CH= CH-CH- 2 3- 1-1K *E 7 t 2- 5- Amino-i, 2, 4- thia diaz ol -3-yl 2-me tho xy imino ace tamido) 3- 3- 3hyd roxymethy lpyr id ini o) 1-propenl 1- 3 -cephem- 4 -carboxyla te (1-1K) (E isomer) A. Diphenylmethyl 2- (5-Axino-1, 2, 4-thiadiazol-3-yl)-2mrethoxyiminoacetamido 1-3- (3-hydroxymethylpyr idinio propenyl ]-3-cephem-4-catboxyl ate iodide CE-isomer) (XII-lK) To a solution of IX-l (E-isorner, 1.79 gr 2.5 mmoles) in'~ ml of CH 3 OHi and 7. 5 ml of CH 3 CN was added 3-hydroxymethylpyridine (545 mg, 5 mroles), and the mixture was stifrrdd at room temperature for 3 hours. The reaction mixture was poured into ethyl acetate (100 ml) With vigorous stirring. The resulting precipitate was collected by filtration, washed with a small volume of ethyl acetate and dried to give 2.06 g (100%) of the title compound XII-l' as a tan powder. Mp. 170-180 0 C (dec.), IR V a (KBr) in cm" 1780, 1725, 1675, 1615, 1530, 1385, 1225, 10 4 0, 750, 700, UJV X Xpa (C 2

H

5 8) in nm (E 1 290 (196).

1 cm NMR 6 (DMSO D 2 Q) in ppm 3.7 (2H, br.s, 3.91 (3H, s, OCH 3 4.70 (2H, s, Py-CH 2

OH),

5.28 (2H, mn, CH 2 -N 5.23 (1H, d, 5.90 (1H, d, 6.34 mn, 3-CH CH), 6.86 d, J=16Hz, 3-CH), 6.89 (1H, s, CHPh 2 7.35 (10H, mn, Ph-H), 7.9-8,9 (4H, mn, Py-H).

S. 7-(2-(5-AMino-1,2,4-thiadiazo-3-y)-2-nethoxYiinoacetamido)-3-(3-(3-ydroxyrnethylPyridilio)-l-propely_1-3ccphem-A-carboxylate (I-1K) (E isomer) A mixture of XII-lK (E isomer, 2.0 g, 2.4 inxoles) and soddium bisulfite (1 g) in 85% ECOOH (10 ml) was stirred for 2 hours at room temperature. The reaction mixture was concentrated to ca. 5 ml under reduced pressure. The oily residue was poured into acetone (100 ml) 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/C 1 8 cartridge (Waters)) to give 283 mg of 1-1K as an amorphous powder. The powder was crystallized from 4N H 2so4 and acetone to give 144 mg of the title compound I-lK as caolorless needles. Mp. 185-188 0 C (dec.).

IR V max (KBr) in cmin 1 1775, )680sh, 1660, 1630, 1225, )1045, 850.

UV X Ma (Phosphate buffer, pH 7) in nm (E 1% 236.5 (283)f 1cm) 275 sh (280), 29 2 .5 33 0) dryness to give a yellow residue, which was triturated with isopropyl ether and filtered to yield 350 mg of the quaternized C Ib t' I t r 6 (D 2 0) in ppm 3.75 (2H, s, 4.18 (3H, s, OCH 3 4.97 (2H, s, Py-CH 2 OH), 5.35 d, J=4Hz, 5.43 (2H, d, CH2-N 5.92 d, J=4Hz, 6.18 (1I, d-t, J=16Hz, J=6.5Hz, 3-CH=CH-), 6.97 (1H, d, J=16Hz, 3-CH), 8.13 (1H, d-d, J=8Hz, J=6Hz, Py-H), 8.60 (1I, d, J=8Hz, Py-H), 8.84 (1Hi, d, J=6Hz, Py-H), 8.90 (1H, s, Py-H).

Example 21 N CONH S

N

2N S OCH3 0 CH=CH-CH 2

ONHCHH

3 CO P I-1M *E 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(Z)-methoxyiminoacetamidol-3-([3-(4-N-methylcarbamoylpyridinio)-1-propenyl]-3cephem-4-carboxylate (1-114) (E isomer) A mixture of diphenylmethyl 7-[2-(5-amino-1,2,"thiadiazol-3-yl)-2-methoxyiminoacetamido -3-(3-iodo-l-propenyl)- 3-cephem-4-carboxylate (IX-l) (E isomer, 450 mg, 0.62 inmole) and 4-N-methylcarbamoyLpyridine (prepared according to the procedure of M. Samejima, Yakugaku Zasshi, 80, 1706 (1960)1 (215 mg, 1.38 mmoles) in acetonitrile (2 ml) 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 'jZ .rmic acid (2 ml) was stirred for 4 hours and then heated at 40 0 C for 30 minutes.

The mixture was evaporated under reduced pressure. The residue was triturated with acetone and the crude product was collected by filtration. The crude product was chromatographed on a column of HP-20 (1.5 x 18 cm) and the column was eluted with ater and aqueous methanol. The methanolic eluate was evaporated under 0 e, reduced pressure and the residue was freeze-dried to give 140 mg of an amorphous -powder, which was further purified by- HPLC (Column: Lichro sorb RP-18, Solvent: 15% CH 3 OH) and the eluate of IHPLC was freeze-dried to give 60 mg of the title product 1-11A. Mp. 180-183 0 C Estimated purity: IR Vmax (KBr) in cm- 1 1760, 1660, 1600.

UJV X mx(Phosphate buffer, pH 7) in nm 230 (22100), 286 (22100).

NMR (D 2 0) in ppm 3.08 (3H, s, CONHCH 3 3.72 (2H, s, 2-H), 4.16 (3H, s, OCtI 3 )r 5.35 (1H, d, 000 Oo6-H), 5.95 (1H, d, J=4.5Hz, 7. 00 OUI_ (1H, d, J=l6Hz, 3-CH), 8.35 (2H, df J-6Hz, pyridine-H), 9.05 (28,r J=J,6Hz, pyridine-H) Example 22 N CONH N* H 1N S OCR 3 cH Cl-=CH-CH1-'jj~ OOHi 0I-1N *E/Z 7/i 7-t2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido)-3t3-(4-carboxypyridinio)-l-2ropenyl)-3-cephem-4-carboxylate (I-lN) To a stirred suspension of isonicotinic acid (340 mg, 2.8 rrmoies) in dry DMF (3.5 ml) was added N,O-bis(trimethylsilyl)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-yl)-2-methoxyiminoacetamido]-3-(3iodo-l-propenyl)-3-cephem-4-carboxylatd (IX-l) (E isomer, 720 mg, 1 mmole) in one portion, and thz red solution was stirred for hours at room temperature. The reaction mixture was added phase column chromatography. The column, which was packed with the packing taken out of a prepPAK-500/C 1 8 cartridge column *i 81 dropwise to a stirred saturated -sodium chloride solution (50 m', containing sodium thiosulfate (150 mg). The yellow precipitate was collected by filtration, washed with water, and dried to obtain 722 mg of a pale yellow powder. The powder (700 mg) and sodium bisulfite (70 mg) were dissolved in 85% formic acid ml), and the solution was allowed to stand at room temperature for 1.5 hours. The mixture was suspended in toluene (50 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 8 g cartridge (Water's System 500), and the column was eluted by water (200 ml). The eluent was fractionated 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 of the title compound I-1N as a yellow powder. E/Z 7/1; 80% pure. Mp. >189 0 C (dec.).

IR V (KBr) in cm 1 1770, 1665, 1600.

max UV 1 (Phosphate buffer, pH 7) in im 227 (22500), max 290 (22100).

NMR 6 (D 2 0 NaHCO 3 in ppm 3.7 (2H, br.s), 4.15 (3H, s), 5.32 (1H, d, J=4Hz), 5.39 (2H, d, J=6Hz), 6.14 (1H, d-t, J=15.5 and 6Hz), 7.03 (1H, d, J=15.5Hz), 8.31 (2H, d, J=7Hz), 8.94 (2H, d, J=7Hz).

Example 23 N C

CONH

i N

C

N N 0 NCH=CH-CH- 2 N S- C 3 0 oe2- 1-10 7-(2-(5-Amino-l, 2, 4-thi ad iazol-3-yl)-2- -me thoxyiminoacetami do) 3-cyclopentenO2Yrid in io)-1-propenyl l-3cephem-4-carboxylate (I-10) (E isomer) A mixture of diphenylmethyl 7-12-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(3-iodo-l-propelyl)- 3-cephem-4-carboxylate CIX-1) (E isomer, 450 mg, 0.62 mmole) and 2,3-cyclopentenopyridine (217 mg, 1.83 mmole) in acetoriitrile (2ml) 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 aci3 (2 ml) was stirred under nitrogen for 3 hours at room temperature and then heated at 40 0 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 cm). The column was eluted with water and 30% aqueous methanol. Evaporation of the methanolic eluate under reduced pressure, followed by freeze-drying afforded 160 mg of an amorphous powder, which was further purified by EPLC (Column: Lichrosorb, Solvent: CH 08). The eluate of HPLC was freeze-dried to give 50 mg of the title product 1-10. Mp. >190 0 C Estimated purity: IR Vma x (KBr) in cm1 1765, 1670, 1600.

CiV Xmax (Phosphate buffer, pH 7) in nm 235 (20000), 283 (25000).

.s 3nun±u ana the mixture was stirred at room temperature for minutes. The solution was filtered and concentrated under h.

NMR 6 (D 2 0 NaHCO 3 in ppm 2.2-2.6 (2H, m, -CH 2 3.1-3.6 (4H, m, -CH 2 3.72 (2H, s, 4.17 (3H, OCH 3 5.33 (1H, d, J=4.5Hz, 5.90 (1H, d, 6.75 (1H, d, J=16Hz, 3-CH), 7.65-8.2 (3H, m, pyridine-H).

Example 24

H

2

N

cooe I-2N 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-(Z)-ethoxyiminoacetamido]-3-[3-(4-carboxypyridinio)-l-propenyl]-3-cephem-4carboxylate (I-2N, E isomer) and 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-(Z)-ethoxyiminoacetamido]-3-3-(4-carboxypyridinio)-l-propenyl]-3-cephem-4carboxylate (I-2N, Z isomer) To a chilled mixture of BSA (1.0 ml, 4.12 mmoles) and isonicotinic acid (506 mg, 4.12 mmoles) was added IX-2 (from Preparation No. 21) il.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 ml) to 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 g) in formic acid 5 ml) was heated at for 1 hour and evaporated under reduced pressure. The residue was triturated with acetone and filtered to give 900 mg of the crude product (E-propenyl isomer:Z-propenyl isomer 2:1).

Separation of the isomers was carried out by HPLC (Column: 84 Lichrosorb, Solvent: 15% CH 3 OH). The faster moving fractions of HPLC were collected, evaporated under reduced pressure and freeze-dried to give the E-propenyl isomer of I-2N (44 mg, yield The slower moving fractions gave the Z-propenyl isomer of 1-2N (32 mg, yield in a similar procedure.

I-2N, E isomer Mp.: >200 0 C (dec.).

IR :Vma x (KBr) in cm 1 1765f 1660, 1620, 1380.

UV X ~max (Water in nm 228 (22200), 291 (23600).

N MR 6 (D 2 0) in ppm 1.45 (3H, t, J=6Hz, CH 2 CH 3.72 s, 4.45 (2H, q, CH 2 CH 3 5.4.0 (1H, d, J=4Hz, 5.90 (1H, d, J=4Hz, 7-H), 7.05 (18, d, J=15Hz, 3-CH), 8.30 (2H, d, J=6Hz, Py-H), 8.95 (2H, d, J=6Hz, Py-H).

I-2N, Z isomer >200 0 C (dec.) I-R Vma (KBr) in cm- 1 1760, 1660(sh), 1620, 1370.

UV Xmax (Phosphate buffer, pH 7) in nm 2 25 22 40 0) 27 (sh, 16000).

NMR :6 (D 2 0) in ppm 1.45 (3H, t, J=7Hz, CH 2 CH 3 3.50 (1H, d, J=17Hz, 3.75 (18, d, J=17Hz, 2-H), 5.38 (1H, d, J=4Hz, 5.95 (18, d, J=4Hzj 6.62 (1H, d, J=llHzf 3-CH), 8.35 (2H, dt J=6Hz, Py-H), 8.92 (2H, d, J=6Hz, Py-H).

collected by filtration, washed witft acetone (2 x 100 ml) and

U-

Example N CONHS iN N SN' \O H 2 N .2 2 H'H 2-NCN 2 CH 2-CH=CH 2 Y 1,> 1-3H 7-[2-(Amino-1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)a c eta m ido I1- 3- 3- 4- ca rb a moyl1py r id injo pr o pe n y 1 3 -c e p hem-4 carboxylate (1-3H) (E isomer) To a solution of 35 mg (0.08 mole) of 7-amino-3-[3-(4carbamoylpyridinio)-l-(E)-propenyll-3-cephem-4-carboxylate hydrochloride in 2 ml of 50% aqueous acetone was added 52 mg of 2-t5-amino-l,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyim ino)acety.

chloride hydrochloride (from Preparation No. 25) and the mixture was adjusted to pH 6.5-7.0 with 2N Na 2 00O 3 The mixture wa.4 stirred at room temperature for 1 hour, acidified to pH 2 with 1lN HCl and concentrated under reduced pr'essure. The residue was chromatographed on a column of HP-20 resin which was el1uted with 300 ml of water and 30% CH 3 OH-H 2 0. Fractions containing the product were combined and evaporated und, r reduced pressure. The residue, 73 mg, was purified by a column of reverse phase carrie which was taken out of a PrepPAK-500/C 18 cartridge (Waters, ml). The column was eluted with water, 5% CH 3 OHt 10% CH 3 OH and CH 3OH, successively. Fractions containing the product were combined and lyophilize-d to afford 26 mg of the title product I-3M. Mp. 160 0 C (dec.).

IR ma in cm- 3400, 1765, 1680, 16050 1400.

UV mrax (Phosphate buffer, pH 7) in nm 226 (24600), 28E (22800).

*i aI u ovuumi DISulrlle (16 g) in 85% HCOOH (160 ml) was stirred at roo.m temperature for minutes and subsequently at 40 0 C for 1 hour under

N

2 The 2* 86 NMR 6 (D 2 0) in ppm 3.75 (2H, s, 5.41 (1H, d, 5.50 (4H, m, CH 2 N CH=CH 2 5.98 (1H, d, J=SHz, 6.20 (1l, m, 3-CH=CH), 7.09 (1R, d, J=17Hz, 3-CH), 8.50 (2H, d, J=7Hz, Py-H), 9.16 (2H, d, J=7Hz, Py-H).

Example 26

S

NC CONH NAS T N C C2 CH=CH-CH CONH2

H

2 N S 02 _2 2C-CH COcP I-4H *E 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)--propargyloxyiminoacetamido -3-[3-(4-carbamoylpyridinio)-1-propenyl)-3-cephem- 4 carboxylate (I-4H) (E isomer) To a solution of 86 mg (0.19 mmole) of 7-amino-3-(3- (4-carbamoylpyridinio)-1-E)-propenyl-3-cephem-4-carboxylate hydrochloride (XXII-H) in 2 mi of 50% aqueous acetone was added 63 mg of 2-propargyloxyimino-2-(5-amino-l1,2,4-thiadiazol-3-yl)acetyl chloride hydrochloride (from Preparation No. 26). The suspension was maintained at pH 6.5-7.0 with 2N Na 2

CO

3 and then stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 2 with IN HC1 and concentrated in vacuo. The residue was diluted with 30 ml of water, neutralized with NaHCO o 3 and filtered. The filtrate was transferred on the top of a column which was packed with reverse phase carrier (30 ml) taken Sfrom a PrepPAK-500/C 18 cartridge (Waters). The column was eluted with water, 5% C 3 OH, 10% CH 3 OH and 20% CH3 OH, successively.

Fractions containing the product were combined and lyophilized to afford 13 mg of the title product I-4H. Estimated purity Mp. 160*C.

IR Vma (KBr) in cm 1 3400, 2120, 1765, 1680, 1610.

max occasional additions of sodium bicarbonate. The reaction was also monitored by tic. After all of Compound XXII-H had been UV X max~ (Phosphate buffer, pH 7) in nm (E) 229 (24000), 288 (21200).

N MR 6 (D 2 0) in ppm 3.78 (2H, s, 5.15 (2H, d, J=i~z, CH 5.0(H dJSz 6-, CHN5. 5.9 d, =56H, 50 (2H, mn, _HN),5.98(Id z 6. 20 1lH, M, 3-CH=CH) 7 .05 IH, d, J=17Hz 3-CH) 8. 50 2H, d, J=7Hz, Py-H), 9. 16 2H, d, J=7Hz, Py-H) Example 27

S

CONH

N

0 N CH=CH-CH 2 -TN/ CONH 2 I 0 7 5-Akmino-i1, 2, 4 -thi ad iaz ol -3-yi -2-cycl open tyl oxyiminoac et amido I- 3- ca rbamoyl py r idin io) -i-pr openyl I- 3-ce ph em- 4carboxylate (1-5H) CE isomer) To a stirred solution of 139 mng (0,31 inmole) of 7-amino- 3 3 -ca r bamoylIpy r idin io)- 1-p rope ny11- 3 ceph em- 4-c ar box yIa t e hydrochloride in 3.5 ml of 50% aqueous acetone in an ice-cooling bath waa added portionwise 120 mng (0.44 inmole) of 1,2,4-thiidiazo-3-yl)-2-cyclopentyioxyiminoacetyI chloride hydrochloride (from Preparation No. 27). The mixture was adjusted to pH 6.5-7.0 with 2N NaCO 3 (0.9 ml) and stirred for 1 hour at 10 0 C. The reaction mixture was acidifi~ed to pH 2 with IN HC1 and evaporeated under reduced pressure. The residue was chromatographed on a column of HP-20 resin (20 ml) and was elUted with 300 ml] of water and 30% CH 3 OH-F 0, successively. Fractions containing the product were combined and concentrated in vacuo.

The residue was treated with 60 ml of acetone to give 111 mg of the title compound I-5H. M1p. 160 0 C (dec. ).Estimated purity form of I-lH (pale yellow amorphous powder; estimated purity

L

88 -l IR V (KBr) cm 3400, 1770, 1680, 1605, 1530.

max UV X (Phosphate buffer, pH 7) in nm 224 (23300), max 286 (24600).

NMR 6 (DMSO-d) in ppm 1.70 (8H, br.s, 4.68 c1H, br.s, 5.05 (1Hi, d, 5.30 (2H, m, CH 2 N 5.67 (1H, d-d, J=5Hz 7Hz, 6.20 (1H, m, 3-CH=CH), 7.08 (1Hi, d, J=17HZ, 3-CH), 8.34 ,2H, d, J=7Hz, Py-H), 9.11 (2Hf d, J=7Hz, Py-H), 9.38 (lHf d, J=7Hz, 7-NH).

Example 28

S

NCON

H

2 C00H S; ICH H=CH-CH 2 I-1P *E 7(2- (5-Amino-1,2,4- thiadiazol-3-yl)-2-methoxyimifloacetamido)-3- [3-(3-carboxymethylpyridinio)-l-propeflY-3-cephem-4-carboxylate (I-lP) (E isomer) A. Diphenylmethyl 7-2-(-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido3-3-[3-C3-carboxymethylpyridinio)-lpropenyl)-3-cephem-4-carboxylate (XII-lP, iodide, E isoer) To a suspension of 3-carbo;ymethylpyridine hydrochloride (0.89 gf 5 mmoles) in 10 ml of CH 2 C1 2 was added N,0-bis(trimethylsilyl)acetamide (4.97 ml, 18 mmoles), and the mixture was stirred at room temperatur6 until a clear solution was obtained. To the solution was added IX-l (1.79 g, mmoles), and the mixture was allowed to stand at room Column, Lichrosorb RP-18, 4x300 mm: Mobile phase, 0.01 M 89 temperature. After 3 hours, 3 ml of CH 3 0H was added to the cooled mixture and the solution was evaporated in vacuo to give an oil which was tril-urated with ethyl acetate to afford 2.28 g of the title compound XII-lP as a tan powder. Mp. 161 0 C (dec.).

IR Vmax (KBr) in cm- 1 1780, 1720, 1675, 1630, 1530, 1385, 1225, 1045, 755, 700.

UV X max (C 2 H 5 OH) in nm (E 1% 295 (188).

I cm IqMR 6 (DMSO D 2 0) in ppm 3.70 (211, br.s, 3.90 (5H1, sf 0C11 3 Py-CH CO), 5.25 (3H1, m, +j

-CH

2 N 5.92 (1H, d, 6.35 (lH, m, 3-CH=CH-)f 6.90 (2IHI d, J=1611z, 3-CH), 6.92 (111, s, CHPh 2 7.35 mt Ph-H), 8.8-9.0 (4H1, m, Py-H).

B. 7-[2-(5-Amino-1,2,4-thiadiazol-3-y1)-2-methoxyimirnoacetamido (3-carboxymethylpyridinio) -l-propenyl cephem-4-carboxylate (E isomer) A mixture of XII-lP (iodide.) (2,28 g) and sodium bisulfite (1.1 g) in 85% HCQQH (10 ml) was stirred at room temperature for 2 hours. The reaction mixture was concentrated to ca. 5 ml under reduced pressure. The oily residue was triturated with acetone (100 ml) to give 1.22 g of the crude product, which was purified by column chromatography (HP-20, 420 ml) to afford 533 mg of the title compound 1-iP from IX-I) as a pale yellow amorphous powder. Mp. 165 0 C (dec.).

ZR ma x (KBr) in cm- 1 1770, 1670o 1600, 1530, 1385, 1140, 1040.

UV Xmx(Phosphate buffer, pH 6.2) in nm (E 1% 23 4 37 4) ,ma1 cm 277sh (390), 290 (402) IIMR (D 2 0 NaHC 3 in ppm 3.78 (2H, s, 3.92 (2H, s, Py-CH 2 CO), 4.22 (3H, s, OCH 3 5.40 (1H, d, J=4Hz, 5.44 (2H, d, J=6.5Hz, -CH 2 N 5.97 (lE, d, J=4Hz, 6.20 (lE, d-t, J=16 6.5Hz, 3-CH=CH)r 7.08 (1H, d, J=l6Hz, 3-CE), 8.12.

(1H, d-df J=8 &i 7Hz, PY-H 5 8.53 (lE, d, J=8Hz, Py-H 4 8.82 (1H, d, J=7Hz, Py-H 6 )f 8.86 (lii, s, Py-H 2 Example 29 s-cH 2 C00 I- I.Q 7.-[2-(5-Amino-l, 2, 4-thiadi azol-3-yl )-2-methoxyiminoacetami do 1-3- 3-(4-carboxymethylthiopyridinio)-l-propenyl]-3-cephem-4carb-oylate (I-lQ) (E isomer) A. Diphenylmeth/l 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido)-3-(3-(4-carboxymethylthiopyridinio)- 1-propenyl!- 3-cephem- 4-ca rboxyl ate (XII-lQ, iodide, E isomer) To a suspension of 4-carboxymethylthiopyridine (0.88 g, mxnoles) in 1.0 ml of CH 2 C1 2 was added NtQ-bis(trimethylsilyl)acetamide (5 ml, 18 mmoles), and the mixture was stirred at room temperature until a clear solution was obtained. To the solution was added IX-2. (t isomer 1.79 g, 2.5 mmo3.es), and the mixture wzz L f C I i.L 91 allowed-to stand at room temperature. After 3 hours, 3 ml of CH3OH was added to the cold mixture and the solution was evaporated in vacuo to give oily ,,esidue which was triturated with ethyl acetate to give 2.43 g of the tite compound XII-lQ (iodide) as a tan powder. Mp. 155 0 C (dec.).

-1 IR max (KBr) in cm 1780, 1720, 1670, 1625, 1525, 1385, max 1225, 1115, 1040, 755, 700.

UV max (C 2

H

5 0H) in nm (E 1 312 (299).

1 cm NMR 6 (DMSO-d 6

D

2 0) in ppm 3.70 (2H, br.s, 3.93 (3H, s, OCH 3 5,07 (2H, m,

CH

2 5.23 (IH, d, 5.90 (1H, d, J=5Hz, 7-H), 6.29 (lH, m, 3-CH=CH), 6.87 (1H, d, J=16Hz, 3-CH), 6.91 (1H, s, CHPh2), 7.35 (10H, m, Ph-H), 7.88 8.58 (each 2H, d, J=6Hz, Py-H).

B. 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-(4-carboxymethylthiopyridinio)-1propenyl]-3-cephem-4-carboxylate (I-1Q) (E isomer) A mixture of XII-1Q (iodide, 2.43 g) and sodium bisulfite (1.1 g) in 85% HCOOH (10 ml) was stirred at room temperature for 2 iours. The reaction mixture was concertrated to ca. 5 ml under reduced pressure. The oily residue was triturated with acetone (100 ml), filtered and dried to give a crude product (1.39 which was purified by column chromatography (HP-20, 20 ml) to afford 577 mg of the title compound I-IQ (39% from IX-1) as a pale yellow amorphous powder.

Mp. 1880C (dec.).

IR max (KBr) in cm"- 1765, 1670, 1625, 1530, 1380, 1110, 1035.

I

UV m (Phosphate buffer, NMR 6 (D 2 0 NaHCO in ppm pH 6.2) in nm (E 234 (459), Cm 310 (678).

3.79 (2H, br.s, 4.10 (2H, s, S-CH 2 4.23 (3H, S, OCH 5.25 (2H, d, d=6.5Hz, CH 2 -N 5.39 (1H, d, J=4.OHz, 5.97 (1H, d, J=4Hz, 6.18 (lH, d-t, J=15.5Hz 6.3Hz, 3-CH=CH), 7.05 (lI, d, J=15.5Hz, 3-CH), 7.84 8.55 (each 2H, d, J17Hz, Py-H).

Example H3CNO/ 'CH=CH-Cfl 2

H

2

N

CbcP I-1A *E/Z 7/1 7- (5-Amino-i, 2, 4-thiadiazol-3-yl )-2-methoxviminoacetamido l 3-(3-(l-methylpyrrolidinio)-l-propenyl l--ehm4-abxlt sulfate (I-lA, sulfate) A. Diphenylmethyl 7 5-Amino-i, 2, 4-thiadiazol-3-yl methxyiminoacetamido)-3-[3-(l-methylpyrrolidinio)l rprapnyl...3-cephem-4-carboxylat_ (XII-lA, iodide) To a cold solution of diphenylmethyl 7-(2-(5-amino- 1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido (3-odopropenyl)-3-cephem-4-carboxylate (IX-l) (from Preparation No. 14) (21.5 g, 30 mmoles) in ethyl acetate (300 ml) was added dropwise a solution of 1-met'ylpyrrolidine (2.55 g, 30 mmoles) in ethyl acetate (0 ml) over a period of 1 hour at -5 to OOC, with stirring, After stirrin5 for an additional 10 minutes, the resulting precipitate Was collected by filtration and washed with aqueous methanol. The methanolic eluate was evaporated under 93 chloroform (200 ml) to give 23.0 g of t.he title compound (IX-lA, iodide), melting at >175*C (dec.).

IR :V max (KBr) in cm- 1 330C 1780, 1730, 1685, 1615.

UV :X max (CC 2

H

5 08) in rim (E 1 2118 (435), 295 (188).

B. Diphenylmethyl 7-(2-(5-Anmino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3-[ 3-C l-methylpv, rrolidinio)-lpropenyl]-3-cephem-4-carboxylate (XII-lA, chloride) The compound (XII-lA, iodide) (23 gf 28.7 mmoles) was dissolved in a mixture of acetone and methanol 230 ml) and applied on "n Amberlite IRA-410 (chloride form, 230 ml) column_ which was pretreated with the same mixed solvent. The column was developed with the solvent and the fractions containing the desired compound were combined ano' concentrated to in 6ily residue, which was triturated with ethyl acetate (300 ml) to yield 17. 9 g (87. of the title compound (XII-lA, chloride) melting at 190*C (dec.).

IR V ma x (KBr) in cm 1 3380, 1780, 1680, 1620.

UV Xx (C H 5 OH) in nm (E 1 220 (369), 290 (232).

max 2 51cm hours at room temperature. The reaction mixture was duutu 94 C. 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-(1-methylpyrrolidinio)-.-propenyll-3cephem-4-carboxylate sulfate (I-1A, sulfate) A mixture of the compound (XII-1A, chloride) (17.8 g, mmoles) in 85% formic acid (178 ml) 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 g of crude I-lA. Concentration of the filtrate and the acetone washings yielded additional 2.95 g of crude I-1A. Two crops of the crude material were combined and extracted with 2N HC1 (1 L and 0.5 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 evaporated in vacuo to an oily residue, which was triturated with isopropanol (200 ml) and acetone (200 ml), successively, to yield 7.09 g of a light yellow powder. This material (6.80 g) was dissolved in water (20 ml) and then subjected to column chromatography over the packing of PrepPAK-500/C 1 8 cartridge (90 ml), using water and 10. aqueous methanol as an eluent. The eluate was collected in fractions with monitoring by HPLC.. [Column, Nucleocil SSC-ODS-262, 8 x 100 mm; Mobile phase, 0.01M phosphate buffer (pH 7.2)/CH 3 OH 90:10; Detection, UV (254 Fraction No. 4 through Fraction No. 10 were combined, evaporated under reduced pressure and lyophilized to give 2.28 g of a yellow powder (E/Z 7/1, 70% pure) [Crop Fraction No. 11 through Fraction No. 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 A portion of Crop 1 (1.0 g) was purified by rechromatography on the packing of PrepPAK-500/C 1 8 cartridge (90 ml). The column was eluted with water and 5% aqueous methanol, successively. The eluate containing the desired compound was concentrated and lyophilized to yield 638 mg (E/Z 7/1, 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 IN sulfuric acid (5 ml). The solution was diluted h acetone (315 mrl), with stirring. The creamy -precipitate was collected by f iltration to obtain 1. 48 g of the title compound (I-lA, sulf ate) CE/Z 80% pure), melting at >185 0 C (dec.).

IR V max> (KBr) in cm1 3380, 3000, 1765, 1675, 1630, 1535, 1390, 1115.

UV X max~ (Phosphate buffer, pH 7) in nm 236 (19900), 291.5 (22500).

H

N LP 6 (D 2 0 NaHCO 3 in ppm 2.36 (4H1, br., N),3.15 (3H1, s, CH 3 iNG 3,62 br., 2-H andN~) 3.83 (l1i, br. r 4.13 (2Hf d, J=BHz, CH (311, s, OCH 3 5.39 (1H1, d, J=4.5Hz, 5.96 (lH, d, J=4.5Hz, 6.00 (111, m, 3-CH=CH) 6 67 (1/8H1, d, J=l OHz 3-CH, cis) 7.04 (7/8H, d, J=16Hz, 3-CU, trans).

FI Prrl 96 Example 32

S

N C CONH

H

2 N

OCH

3 0

CH=CH-CH

2 -N (CH 3 )3 I-1D *E/Z 10/1 Amino-1,2,4-thiadiazol-3-yl)-2-methoxyimin0acetamidoI- 3-[3-trimethylammonio-l-propenyl)-3-cephem-4-carboxylate (I-ID) A. Diphenylmethyl 7-[2-(5-amino-l,2,4-thiadiazol-3-yl)-2 methoxyiminoacetamido-3-(3-trimethylammonio-l-propenyl)- 3-cephem-4carboxylate (XII-lD, iodide) To a solution of 13.0 g (19 mmoles) of diphenylmethyl 7- 2-(5-amino-1,2,4-thiadiazol-3-yl )-2-methoxyiminoacetamido-3- (3-iodopropenyl)-3-cephem-4-carboxylate (IX-l, from Preparation No. 10) in 38 ml of dry ethyl acetate was added 1.75 ml (19.1 mmoles) of 1.1N trimethylamine in ethyl acetate at -51C, and the mixture was stirred for 1 hour at -5 0 C. The resulting precipitate was filtered off, washed well with CHCl 3 and dried to give 12.5 g of the title compound (XII-lD) as the iodide.

I.R V (KBr) in cm 1 3300, 1765, 1720, 1665.

UV Xmx (C 2

H

5 OH) in nm 300 (18400).

B. Diphenylmethyl 7-12-C5-Amino-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3-(3-trimethylammonio-l-projeyl)- 3-cephem-4-carboxylate (XII-lD, chloride) The iodide (XII-lD, 12.5 g) was dissolved in 60 ml of

CH

3 QH-acetone and passed through a column of ion-exchange resin [IRA-410 125 mll. The column was eluted with 300 ml of CH 3 OH-acetone and the eluate was evaporated in vacuo and triturated with 300 ml of isopropyl-ether to afford 10.4 g of the quaternary salt (XII-lD, chloride).

i-we j j-uieL:.4-propenyi isomer 2:1).

Separation of the isomers was carried out by HPLC (Column: 97 -1 IR v (KBr) in cm 3300, 1765, 1710, 1665, max UV max (C 2 H OH) in nm 298 (15100).

C. 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[ 3-trimethylamnonio-l-propenyl ]-3-cephem- 4-carboxylate (I-ID, sulfate, E isomer) A solution of 10.4 g (16.0 mmoles) of XII-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 eluted with water (1000 ml), CH3OH (200 ml, and 30% CH3OH (150 ml), successively. Fractions concaining the desired product were combined and concentrated under reduced pressure. The residue was purified by reverse phase chromatography. The column was packed with a packing obtained from a PrepPAK-500/C 1 8 cartridge (Waters, 200 ml).

Elution with water (600 ml) and 30% CH30H (200 ml), successively, followed by concentration of fractions containing the desired product gave 2.52 g of the title compound. A solution of the zwitterionic product (1.5 g) in 1N H2So 4 (5 ml) was added portionwise to 300 ml of acetone and the resulting precipitate was filtered and dried. Yield of I-1D sulfate was 1.42 g The ratio of E/Z was approximately 10/1 based on HPLC.

IR max (KBr) in cm 1 3380, 1765, 1665.

UV max (Phosphate buffer, pH 7) in nm 237 (19500), 293 (22400).

98 NMR (D C 2 0) in ppm 3.25 (9H, s, N -CH 3 3.94 (2H, S, 4.14 (2H, d, J=78z, CH 2 N 4.23 (3H, s, O-CH 3 5.42 (18, d, 6.00 (1H, d, J=4.5Hz, 7-H) 6. 23 (1H, d-t, J=7 16Hz, 3-CH=CH), 7.23 (1H, d, J=l6Hz, 3-CH) Example 32

S

C-C ONH N\ C 'YCONH 2 OCH 0o 2 7-[2-(5-Amino-1,2,4-tbiadiazol-3-yl)-2-methoxyiminoacetamido]- 33-C 4-carbamoylpyridinio)-l-propenyl]-3-cephem-4-carboxylate (I-18, E isomer) To a mixture of 7-amino-3-(3- (4-ca rbamoylpyridinio l-(E)-propenyl)-3-cephem-4-carboxylic acid hydrochloride (XXII-H, 397 mg, 1 mmole) and NaHCO 3 (168 mg, 2 mmoles) in aqueous DMF (water, 3.5 ml and D14F, 7.5 ml) was added benzotriazol-l-yl-2- (5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetate (479 mg, mxnoles) (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 ml 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 nH 6.8 with NaHCO 3 1 treated with decolorizing carbon, concentrate~d to ca. 1 ml and seeded with a few pieces of crystalline 1-18. After stirring overnight, the crystalline precipitate was collected by filtration to afford the title compound I-111 (zwitterion form). Yield 83 mg Mp.

>165 0 C Physico-chemical data of this product were identical to those of the compound in txample 99 Preparation No. 1 Diphenylmethyl 7-(2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido ]-3-chior omethyl-3-cephem-4carboxylate (IV-l) To a stirred suspension of 2-(5-amino-,2,4-thiadiazol- 3-yl)-2-methoxyiminoacetic acid (111-1) (2.1 g, 10 mmole) in dry CH 2 Cl 2 (50 ml) was added Pci 5 (2.09 g, 10 mmole) at -301C, and the mixture was stirred for 20 minutes at -15 to -20 OC. To the above acid chloride solution was added a solution of diphenylme thyl 7-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (11) (4.5 g, 10 mmole) in CH 2 Cl 2 (50 ml) containing N,O-bis-(trimechylsilyl)acetanide (10 gr 50 mmole) at -30 *C.

After stirring at -10 0 C for 1 hour, the mixture was concentrated to remove the CH 2 Cl 2 and diluted with ethyl acetate (200 ml).

The mixture was washed with 10 aqueous NaHCO 3 (2 x 40 ml) H 2 0 (2 x 20 ml) and brine (10 ml), successively, and dried over MgSO 4 The solvent was evaporated in vacuo and the resulting oily residue (10 g) was dissolved in CHC1 3 (20 ml) and chromatograpihed on a silica gel (Wako, gel C-200, 100 g containing 10 ml of 1/1.5 M pH 7 phosphate buffer) using 1 3 CH 3 OH-CHCl 3 Fractions containing the title compound were evaporated to give 5.7 g (95 of IV-1 as a yellow amorphous powder. M. p. >140 0

C

(dec. IR :v r cm- 3300, 1780, 1720, 1680, 1620.

ma x UV X EtOH nm 245 (1800), 280 (9900).

ma x 14MR 6 DMSO-d 6 3.53 (2H, ABq, 3.94 (3H, s, OCH 3 4.42 PPM (2H, s, 3-CH 2 5.22 (iH, d, J=4.5, 5.92 (iH, d-d, J=4.5 6, 6.93 (1H, s, CHPh 2 7.36 (iOH, m, Ph-H), 8.1 (2H, br-s, NH 2 9.58 (iN, d, J=6, 7-NH).

IR V max~ (KBr) in cm 1 3400, 2120, 1765, 1680, 1610.

100 Preparation No. 2 Diphenylmethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetarnido)-3-iodomethyl-3-cephem-4-carboxylate (V-1) A mixture of IV-1 from Prepa:,ation No. 1 (5.7 g, mmole) and Nal (4.3 g 29 mmole) in dry acetone (50 ml) was stirred for 5 minutes at room temperat'ure. The mixture was concentrated under reduced pressure anid the resulting oil was shaken with a mixture of ethyl acetate (100 ml) and H20(10 ml).

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 as a yellow amorphous powder melting t 120 0

C

(dec.).

IR v r cm- 3300, 1780, 1725, 1680, 1620.

ma x UV X tHnm 245 (17000), 282 (12000).

ma x NMR 6 DMd 6 3.72 (2H, ABq, 3.94 (3H, sr OCH 3 4.23 PPM (2H, s, 3-CH 2 )e 5.21 (1H, d, J=4.5, 5.89 (lHr J=4.5 6, 6.94 (11H, s, CHPh 2 7.35 (10H, mn, Ph-H), 8.12 br-s, NH 2 9,65 (1H, d, J=6, 7-NH).

Preparation No. 3 Diphenylmethyl 7-[2-(5-Amino-1,24-thiadiazo--3-yl)-2me thoxyiminoacetamido I-3-tr iphenylphosphonioriethyl-3-cephem-4carboxylate iodide (VI-l) A mixture of V-1 from Preparation No. 2' (690 mng, 1 mirole) and triphenyiphosphine (786 mng, 3 inmole) in. ethyl acetate ml) was stirred overnight at room Eemperature. The solid which separated was collected, washed with ethyl acetate (2 X purity 101 ml) and dried to give 950 mg (100 of the phosphonium iodide VI-l. M. p. 186 0 C IR v r cm- 33Uu, 1780, 1710, 1680, 1610.

max UV ritO m 268 (15000), 275 (13000), 300 (7300).

max N 1R a DM~SO- d 6 3.52 (2H, br-s, 28), 3.94 s, OCH 3 5.34 pp:-n (18, d, J=4.5, 5.9 M, 6.3 (18, 7.3 (10H, m, Ph-H), 7.8 (15H, m, Ph-LI).

Preparation No. 4 Diphenylmethyl 7-[2-(5-Amino-1,2,4-thiadia8.ol-3-yl)-2-methoxyiminoacetamido]-3-L (triphenylphosphoraniylidene)met',l),-3cephem-4-carboxylate (VII-1) A mixture of VI-1 from Preparation No. 3 (952 mg, 1 mmole), Amberlite IRA-410 (08- form, 500 mg) and N NaOH (4 ml) in CH 2 C1 2 (10 ml) was stirred for 1 hour at room temperature. The mixture was filtered and the separated organic layer was dried over MgSO 4 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 of the title compound VII-1. M.p. >180 0 C (dec.).

IR K cm- 3400, 1750, 1630.

max UV X EtOH rim 268 (12000), 276 (10000), 384 (23000).

max was obtained. To the solution was added IX-l (1.79 g, rnxoles), and the mixture was allowed to stand at room 102 Preparation No. Diphenylmethyl 7- [2-C 5-Awino-., 2, 4-tiadiazol-3-y -2--me thoxyiminoacetamido 3-chloro-1--propen-1-yl -3-cephemn-4-carboxylate CVIII-l) To a solution of VII-l from Preparation No. 4 (6.9 g, 8.4 mmhole) were added MgSO 4 (3 g) and 40% chioroacetaldehyde (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 g containing 10 ml of 1/1.5 M phosphate buffer) column by using CHCl 3 and CH C1 3 containing CH 3OH.

Fractions containing the desired product (0.5 1% CH 3 OH-CHCl 3 were evaporated in vacuo to give 1.6 g of the title compound VIII-l as a yellow amorphous powder, which was a mixture of the Z and E isomers with respect to the chioropropenyl moiety-~ Z/E=2/l, by nmr M.P. >130 0 C (dec.).

IR V Krcm- 3300, 1780, 1725, 1680, 1620.

max UV X EtOH nm 240 (20000), 286 (12000).

max NMR 6 DMSO-d 6

+D

2 O0 ppm 3.56 3.8 Cm, 2-H) 3.94 3H, s, 0CH 3 4.16 Cd, J=7.5, CH 2 Cl), 5.26 (1H, d, 5.87 (TH, d, J=4.5, 7-H), 6,28 (2/3H, d, J=11, 3-CE cis-H), 6.72 (1/311, d, J=16f 3-CH trans-H), 6.81 (2/311, s, CHlPh 2 6.92 (1/3H, s, CHPh 2 7.4 (10H, m, Ph-H).

I

103 Preparation No. 6 piphenylmethyl 7-Benzylideneamino-3-[(triphenylphosphoranylidene)methyl]-3-cephem-4-carboxylate (XVI) To a solution of diphenylmethyl 7-benzylideneamino-3- ((triphenylphosphonio)methyl]-3-cephem-4-carboxylate iodide (XV) [prepared according to the procedure of Japan published patent application (Kokai) 56-86187 (7/31/81)] (60 g, 70 mmole) in

CH

2 C1 2 (350 ml) were added N NaOH (140 ml) and Amberlite IRA-410 (OH form, 35 g) at 5 0 C. The mixture was stirred for 1 hour at 0 C and filtered. The organic layer was separated, dried over MgSO 4 concentrated to ca. 100 ml of volume and precipitated with ethyl acetate (500 ml). The resulting yellow solid was collected by filtration and dried in vacuo to afford 48 g of the title compound XVI, melting at 195-8 0 C (dec.).

IR KBr cm- 1 1770, 1620.

max Preparation No. 7 Diphenylmethyl 7-Benzylideneamino-3-(3-chloro-l-propen-l-yl)-3cephem-4-carboxylate (XVII) To a stirred solution of XVI from Preparation No. 6 (2.9 g, 4 mmole) in a mixture of CH 2 C1 2 (40 ml) and H 2 0 (10 ml), was added anhydrous chloroacetaldehyde (800 mg) at room temperature. To the mixture was added additional 800 mg of chloroacetaldehyde in three portions over 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 4 Evaporation of the solvent gave a red oil which was dissolved in a mixture of ethyl acetate and isopropyl ether 80 ml). The solution was washed with saturated aqueous NaHCO 3 (10 ml) and H 2 0 (10 ml), successively.

After drying over MgSO 4 removal of the solvent afforded 3,3 g of yellow oil. A solution of the oil in CH 2 C1 2 (50 ml) was filtered temperature until a clear solution was was added IX-1 (E isomer 1.79 g, 2. 5 mxnoles), and the mixture was 104 with aid of silica gel (12 g, Wakogel C-200) containing 1/1.5 M phosphate buffer (1.2 ml, pH 6.4) and the silica gel was washed with CH 2

C

2 (50,ml). The filtrate and washing were combined and evaporated to dryness. The residue was triturated with n-hexane to give 1.7 g of the title compound (XVII) as a yel2ow powder. The nmr spectrum indicated that the chloropropenyl moiety h~ad the Z configuration. M.p. ;,50 0 C (dec.).

1, VKBr cm- 1 3400, 1775, 1720, 1630.

ma x UV: XEO nm ma x M~R ~D MS0-d6 ppm 253 (11000), 258 (11000), 265 (10000), 273 (8300), 281 (7000), 290 (6300).

3.63 (2H, br-s, 4.0 (2H, m, CH 2 Cl), 5.42 (2H, m, 6-H 3-CH=CH), 5.72 (lE, di J=.4.5f 6.27 (1H, d, J=11, 3-CE), 6.85 (1li, s, C.4'h2 7, 33 (l10H, m, Ph-H) Preparation of anhydrous chloroacetaldehyde Anhydrous calcium chloride was added to a chilled solution of 50% aqueous chloroacetaldehyde (50 ml), with stirring, to separate it into two layers. The chloroacetaldehyde hydrate l-ayer ()(the upper layer) was separated and diluted with CHCl 3 (100 ml), mixed With MgSO 4 (20 heated to reflux for 5 minutes, and filtered. The solvent and water were removed azeotropically 56-64 0 C) and the residue was distilled to give anhydrous chioroacetaldehyde b.p. 70-826C/760 mm, IR: Vfilm c In 1720.

max R.P. Kurkjy, E. V, Brown, J. Amer, Chem. Soc., 741 5778 (1952).

S. Trippett. D, M. Walker, j. Chem, Soc., 1961 1266.

IR v x (KBr) in cm 1765, 1670, 1625, 1530, 1380, 1110, 1035.

e C, 105 H. O. House, V. K. Jones, G. A. Frank, J. Org. Chem., 29, 3327 (1964).

Preparation No. 8 Diphenylmethyl 7-Amino-3-(3-chloro-1-pZropen-1-yl)-3-cephem- 4 carboxylate (XVIII) 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 CH 3 OH (10 ml) containing acetic acid (0.25 ml), at 5 0 C. After stirring for 30 minutes at 5 0 C, the mixture was concentrated to remove the CH 3 OH and then ethyl acetate ml) was added. The ethyl acetate solution was washed with H20 (2 x 5 ml), saturated aqueous NaHCO 3 (5 ml) and brine (5 ml), successively and dried over MgSO 4 Evaporation of the solvent gave 145 mg of the title compound XVIII (Z isomer) as a yellow powder. M.p. >100 0 C (dec.), R vKBr cm- 1 3400, 1770, 1720.

max UV EtOH nm 252 (3700), 258 (3800), 260 (4000), 274 max (4000), 285 (4000).

Preparation No. 9 Diphenylmethyl 7-(2-(5-Amino-1,2,4-this "'azol-3-yl)-2-methoxyiminoacetamido]-3-(3-chloro-l-propen-1-y1)-3-cephem-4carboxylate (VIII-1) A mixture of 2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetic acie (111-1) (10,1 g, 50 mmole) and PC15 (10.4 g, 50 mmole) in dry CH 2 Cl 2 (100 ml) was stirred at -7 to -15 0

C

for 2 hours. The clear solution was poured into n-hexane (500 ml) to give a precipitate. The organic layer was discarded by 106 decantation and th e remaining solid was triturated with n-hexane (100 ml). The yellow precipitate was collected by filtration and dried in vccuo to give 12.5 g of the acid chloride, melting at 80 0 C (dec.).

IR V nujol cm- 1 1770.

max Tr-e acid chloride (25 mg, 0.1 mmole) Tas added~ to a solution of XVIII (Z i. omer) I:om Preparation No. 8 (44 mg, 0.1 mmole) i Iry CH 2 Cl 2 (5 ml) it room temperature, with stirring.

After 30 minutes, the mixture was concentrated under reduced pressure and diluted with ethyl acetate (20 ml) and saturated aqueous Nai-fCO 3 (5 ml). The organic layer was washed with saturated aqueous NaHCO 3 (5 ml), brine (5 ml), 10% HCl (5 ml) and br,'.ne (5 ml). The solvent was dried over MgSO 4 and th 'en evaporated to dryness to give the product as a yellow foam. Thefoam was purified by silica gel (Wakogel C-200f 1 g, containing C,.1 ml of 1/1.5 M phosphate buffer F-fI 6.4) column~ chromatography by elution with CH2 2 C1 2. H3O (100 to give 31 mg of the title compound VIII-l (Z isomer) as a yellow powder. M.p, >150 r -lc.) IR:V Krcm -13400t 1775, 1720, 1675, 1630.

ma x UV :X EtOH nm 240 (17000), 280 (10000).

ma x tIMR MOdt 3.6 (2H, m, 3.92 (31H, s, O-CH 3 4.0 (2H, PPM mr CH 2 Cl), 5.27 (2H, m, 6-H 3-CH=CH), 5.83 (lIH, d-d, J=4.5 10, 6.25 (1H, d, J=ll, 3-CH)f 6.03 (lHosf CHPh 2 7.33 (10H, m, Ph-H), 8.0 (21, br-s, NH 2 9.57 ClHr d, J=10, 7-NH).

107 Preparation No. DRiphenylmethyl 7-(2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido 1-3- (3-iodo-J.-propen-1-yl )-3-cephem-4-ca rboxylate (Ix-l) A solution of VIII-l from Preparation No. 5 (Z/E=2/l, 480 mg, 0.77 mmTole) in dry acetone (10 ml) 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 acetati (50 ml) and water (10 ml). The upper layer was washed with 10% w/v aqueous sodium thiosulfate solution (10 ml) and brine (10 ml) successively, and dried over MgSO 4 Evaporation of the solvent gave 540 mg of the title compound IX-l as a reddish amorphous solid, melting at >120 0 C (dec.).

IR v r cm- 3300, 1780, 1720, 1680, 1620.

max UV X tHnm 240 (21000), 290 (12000).

max N 1R 6 DMOD2 03 .6 7 2 H rri 2 H) 5. 29 1 H d r J 5, 6 ppm 5.95 (l1H, d, J=4.5, 6.27 d, trans), 6.87 6.96 (each 1/2H, s, CIIPh 2 7.4 (10H, m, Ph-H).

sulfuric acid (5 ml). The solution was diluted h acetone (315 108 Preparation No. 11 Diphenylmethyl 7- [2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido-3-(3-iodo-l-propen-1-yl)-3-cephen-4-carboxylate A mixture of VIII-1 (Z isomer) from Preparation No. 9 9 mmole) and N-I (4 g, 27 mmole) in dry acetone (100 ml) was stirred for 1.5 hours at room temperature. The mixture was evaporated and the resulting oil was diluted with ethyl acetate ml). The ethyl acetate layer was washed with 10% W/v aqueous sodium thiosulfata solution (10 ml) and H 2 0 (10 ml). Removal of the dried (MgSO solvent gave a yellow oil, which was solidified by trituration with isopropyl ether. Filtration of the precipitate gave 4.3 g of the title compound IX-1 as the E isomer. M.p. >165 0 C (dec.), KBr -1 IR v cm 3400, 1780, 1725, 1680, 1610.

max UV XEtOH nm 240 (18000), 297 (11000).

max DMSO-d +D 0 NMR 6 MSO-d6 +D 2 3.90 (3H, s, OCH 3 5,25 (1H, m, 5.95 ppm (1H, m, 6.72 J=16, 3-CH trans), 6,96 s, CH-Ph 2 7.4 (10H, m, Ph-H).

Preparation No. 12 Benzhydryl 7-Amino-3-[3-chloro-1-propen-1-yl-3-cephem-4carboxylate (Z-isomer) (XVIII) Compound XVIII is the common intermediate utilized in Reaction Schemes lb and 1c.

A. Benzhydryl 7-Benzyl ideneamino-3-triphenylphosphonip hyl-3cephem-4-carboxylate chloride (XV) r 109 To a suspension of benzhyd'ryl 7-amino-3-chloromethyl-3cephem-4-carboxylate hydrochloride (II hydrochloride) (200 g, 0.44 mole) in CH 2 C1 2 (940 ml) was added 1 N NaOH (440 ml) at room temperature. The mixture was shaken for 10 minutes and the organic layer was separated. To the organic layer were added MgSO 4 (75 g) and benzaldehyde (51 g, 0.48 mole) and the mixture was allowed to stand for 3 hours. The reaction mixture was filtered and the insolubles were washed with CH 2 C1 2 (200 ml). To the combined filtrate and washings was added triphenylphosphine (126 g, 0.48 mole). The mixture was concentrated to about 400 ml and allowed to stand 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 g M.p. 185-190 0 C (dec.).

IR v K B r cm 1780, 1720, 1630.

max UV CH 2 C1 2 nm 260 (24100).

max B. Benzhydryl 7-Benzylideneamino-3-[ (triphenylphosphoranylidene)methyl]-3-cephem-4-carboxylate

(XVI)

A mixture of XV (322 g, 0.42 mole) and 5 N Na 2

CO

3 (252 ml) in CH 2 C12 (1.6 L) was stirred vigorously for 15 minutes at room temperature. The organic layer was separated, dried over MgSO 4 and concentrated to about 500 ml of volume. The concentrate was added to acetone (1 with stirring, to give a light yellow crystalline powder which was collected by filtration to yield 237 g of XVI, melting at 195-198°C (dec.).

and triturated with 300 ml of isopropyi-etner to arrLuu Lu.of the quaternary salt (XII-1D, chloride).

110 KBr -1 IR v cm 1770, 1620.

max UV CH 2 C 2 nm 254 (23000), 389 (22000).

max CDC1 NMR 6 3 2.56 3.16 (2H, ABq), 5.00 (IH, d, J=4 Hz), 5.23 ppm (1H, d, J=4 Hz), 5.47 (1H, d, J=22 Hz), 6.95 (lH,.

7.2-7.8 (30H, 8.55 (1H, s).

C. Benzhydr yl 7-Amino-3- chloro-l-propen- l-yl]-3-cephem- 4carboxylate hydrochloride (Z isomer) (XVIII Hydrochloride) To a refluxing solution of XVI (214 g, 0.294 mole) and N,O-bis-(trimethylsilyl)acetamide (40 ml, 0.15 mole) in dry

CH

2 Cl 2 (2.9 L) was added dropwise, with stirring, a 50% solutionof chloroacetaldehyde (93 g, 0.59 mole) in CHCl 3 over a period of minutes. After standing for 30 minutes, the mixture was concentrated to dryness. To the residual oil were added CH 2 Cl 2 Girard Reagent T (99 g, 0.59 mole) and 10% aqueous HCI (300 ml), and the mixture was stirred for 1 hour at room temperature. The organic layer was washed with water (200 ml) and a saturated NaCl solution (200 ml), dried over MgSO 4 treated with charcoal (5 g) and filtered. The filtrate was cooled to -10 0

C

and treated with 1 N HCI in CH30H (300 ml). The mixture was stirred for 30 minutes at room temperature and concentrated to about 300 ml. The concentrate was diluted with ethyl acetate (400 ml) and seeded with a few crystals of XVIII hydrochloride.

After 2 hours the separated crystals were collected by filtration, washed with ethyl acetate (200 ml) and dried in vacuo to give 74 g of the title compound XVIII as its hydrochloride, melting at >185 0 C Pale yellow needles.

K~r IR V cm 2830, 17 80, 17 max UV EtX nm 286 (8800).

max NMR 6

MSO-

6 3.73 (2H, br, s, 3.97 (2H, m, CH 2 Cl,52 PPM (18, d, J=4.5 Hz, 5.37 (1H, d, J=4.5 Hz, 5.77 (1H, m, 3-CH=CH), 6.45 (1H, d, J=11 Hz, 3-),68 (1H, s, CHPh 2 7.33 (10H, br, s, Ph-H).

Anal. CaJlc'd for C H N 0SC1'C:C 78;H 23 21 2 3 1 C 78;H N, 5.87; S, 6.72; Cl, 14.85.

Found: C, 57.62; Hr 4.5,3; N, 5.70; S, 6.64; Cl, 14.89, Preparation No. 13 Benzhydryl 7- (5-Amino-i, 2, 4-thiadiazol-3-y1 thoxyiminoacetamido]-3-[ 3-chloro-l-propen-1-yl-3-cephem-4-ca koxylate (Z isomer) (VIII-l) To a stirred solution of XVIII (Z isomer) (20 g, 42 mmole) in CH 2 C1 2 (420 ml) containing N,O-bis(trimethylsilyl)acetamide (34 ml, 125 mmole) was added 2-(5-amino-1,2,4thiadiazol-3-yl )-2-methoxyimi noacetyl chloride hydrochloride (15.2 g, 59 mmole) in three portions over a period of 30 minutes )at -10 to 0 0 C. The mixture w,.s stirred for 30 minutes at 0-5 0

C

and concentrated under reduced pressure. !,he residual brown oil was dissolved in ethyl acetate (420 ml) and the solution was washed successively with saturated aqueous NaHCO 3 (3 x 15 ml), saturated aqueous NaCl (15 ml), 10% HCl (15 ml) and saturated aqueous NaCl (15 ml), and concentrated to about 50 ml of the volume. To the concentrate was added n-heptane (200 ml) to give 112 28.5 g (90% pure) of the title compound VIII-l (Z-isomer) as a colorless powder. M.p. >150 0 C (dec.).

IR :v r cm- 3400, 1780, 1720, 1680, 1620.

max UiV X EtHnm 240 (20000), 283 (12000).

ma x NMR 6 acetone-d 6 3.6 (2H, mn, 3.95 (3H, s, 0CH 3 ppm (2H, mn, CH 2 Cl), 5.32 (1H, d, J=4.5 Hz, 5.62 (1H, in, 3-CH=CH), 6.03 (lH( d, Hz, 6.32 (1H, d, 3=i1 Hz, 3-CH), 6.87 (1H, s, CHPh 2 7.33 (10H, br, s, Ph-H).

Preparation No. 14 Benzhydryl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-nethoxyiinoacetainido]-3-[3-iodo-l-propen-1-yl]-3-cephein-4-carboxylate (E isomer) (IX-l) A mixture of VIII-l (Z isomer) (28.5 g, 90% pure) and sodium iodide (19 g) in dry acetone (420 ml) was stirred for minutes at room temperature and allowed to stand at 5 0 C for 2 hours. The mixture was concentrated under reduced pressure. To the residue were added ethyl acetate (420 ml) and 10% w/v aqueous sodium thiosulfate solution (30 ml), and the mixture was shaken.

The organic layer was washed with water (30 ml), dried over McgSO 4 and evaporated to about 50 ml of volume. The concentrate was diluted with n-heptane (200 ml) to yieJ.' 30.6 g (95% pure) of the title compound IX-1 (E isomer) as a yellow powder, melting at >120 0 C (dec.).

113 IR :VKrc-13400, 1780, 1725, 1680, 1620.

max UV :X EtOH rim E) 306 (15000) ma x N MR 6 acetorie-d 6 PPM 3.71 (2H, m, 3.97 (3H, s, 0CH 3 4.9 (2H, d, J=8 Hz, CH 2 I1), 5.26 (1H, d, J=4.5 Hz, 6.03 (1H, d-d, J=4.5 8 Hz, changed to doublet J=4.5 Hz by D 2 0, 6.32 (1H, d-t, 8 Hz, 3-CH=CH), 6.79 (lHf d, J=15 Hz, 3-CH)f 6.98 (1H, s, CHPh 2 7.35 (10H, m, Ph-H), 7.63 (2H, br, s, disappeared by D 2 0, NH 2 8.52 (1H, d, J=8 Hz, disappeared by D 20, 7-NH).

Preparationi No. Benzhydryl 2-(5-Amirio-l,2, 4-thiadiazol-3-yl )-2-methoxyiminoacetamido]-3-(3-(4-carbamoyl-l-pyridinio)-l-propean-1-y11-3caphem-4-carboxylate Iodide (E isomer) (XII-lH) To a suspension of IX-l (E isomer) (30.5 g) arid isonicotinamide (26 g, 212 mmole) in CH 3 CN (120 ml) was added CH 3 OH (100 ml) until the m'ixture became clear. The solution was stirred for 2 hours under nitrogen atmosphere at room temperature arid concentrated to about 100 ml under reduced pressure. The residual semi-solid was triturated with isopropyl ether (200 ml).

The solvent was removed by decantation arid the residual yellow powder was washed with a mixture of isopropyl ether and CH 3

OH

3/1 120 ml) The powder was collected by filtration arid dried in vacuo to give 36 g (75% pure estimated by HPLC) of the title compound XII-lH (E isomer) as a light yellow powder melting at 15 0 0 C (dec. which separated was collected, washed with ethyl acetate (2 x F~n~P~ IR KBr cm- 1 3300, 1780, 1720, 1680, 1620.

max UV X E t OH nm (El

H

282 (170).

max 1 cm NMIR 6DMSO-d 6 ppm 3.72 (2H, m, 3.90 (3H, s, OCH3), 5.25 (3H, m, 6-H CH 5.9 (1H, d-d, J=4.5 8 Hz, changed to a doublet J=4.5 Hz by D 2 0 addition, 6.35 (1H, m, 3-CH=CH), 6.89 (1H, s, CHPh 2 6.9 (1H, d, J=16 Hz, 3-CH), 7.35 m, Ph-H), 8.06 (2H, br, s, disappeared by D 2 0,

NH

2 8.21 (2H, br, s, disappeared by D 2 0 addition, NH 2 8.36 9.07 (each 2H, d, J=6 Hz, Py-H), 9.57 (1H, d, J=8 Hz, disappeared by D 2 0 addition, 7-NH).

Preparation No. 16 Benzhydryl 7-Benzylideneamino-3-[3-chloro-l-propen-l-yl]-3cephem-4-carboxylate (XVII) (Z isomer) To an ice-cooled mixture of the crystalline 7-aminocephem intermediate XVIII (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 ml, 28 mmole) over a period of 20 minutes, to maintain the temperature of the reaction mixture below 10 0 C. The mixture was stirred with cooling for another 15 minutes, and the organic layer was separated, washed with saturated aqueous sodium bicarbonate (100 ml x 2) 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. This procedure was repeated 3 times, and the mixture was monitored by reverse phase tlc to confirm that all of the starting 7-amino- ~1 115 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 which was used for the next step without purification.

KBr -1 IR v cm 1 1780, 1725, 1635.

max UV XCH2 C 2 nm (E 257 (400).

max 1 cm NMR 6 CDC3 6.18 (1H, d, J=ll Hz).

ppm Preparation No. 17 0 Benzhydryl 7-Benzylideneamino-3-[3-(4-carbamoyll--pyridinio)-lpropen-l-yl]-3-cephem-4-carboxylate Iodide (XXI-H) (E isomer) To a chilled mixture of the 3-chloropropenylcephem XVII (Z isomer) (16.4 g) in acetone (5 ml), was added dropwise a solution of sodium iodide (6.3 g, 42 mmole) in acetone (30 ml) over 10 minutes under nitrogen atmosphere, and the mixture was stirred at room temperature. The reaction was monitored by the ratio of uv absorption [E 1 (255 nm)/E 1 (320 When 1 cm 1 cm the ratio reached below 1.30 (after 45 minutes), the mixture was diluted with carbon tetrachloride (400 ml), 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 was treated with a small amount of charcoal and diatomaceous earth, and filtered. The filter cake was washed with a 1:1 mixture (100 ml) of methylene chloride and carbon tetrachloride. 2o the combined solution of the filtrate and washings, was added a solution of isonicotinamide (3.5 g, 28.7 mmole) in dimathylformamide (20 ml) and the mixture was concentrated under :educed pressure. The concentrate was allowed to stand at room temperature for 1.5 hours and washed with isopropyl 116 eth.er (100 ml x The residual brown semi-solid was dissolved in methylene chloride (50 ml) and the solution was added dropwise, with stirring, to ethyl acetate (1.5 The resulting precipitate was collected by filtration and washed with ethyl acetate (200 ml). After drying over phosphorous pentoxide in vacuo, 17 g of the title compound XXI-H (E isomer) was obtained.

Yellow amorphous powder. M.p. 150-155 0 C Estimated purity 80% by nmr.

KBr -1 IR V cm 1 1775, 1725, 1690, 1635.

max UV hCH 2 C1 2 nm 258 (335), 298 (255).

max 1 cm NMR 6 DMSO-d 6 NMR 6 6 ppm 3.4-3.8 (2H, 5.35 (2H, 5.41 (1H, d, J=4 Hz), 5.73 (1H, d, J=4 Hz), 6.93 (1H, s), 6.97 (1H, d, 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- 4-carbamoyl-l-pyridinio)-l-propen-l-) 3-cephem-4carboxylate (XXII-H) (E isomer) To a suspension of the quaternized cephem XXI-H (17 g) in 8i% formic acid (25 ml) was added dropWise concentrated hydrochloric acid (5 ml), and the mixture was stirred at room temperature for 1.5 hours and treated with a small amount of Scharcoal. The mixture was filtered and washed with 85% formic acid (5 ml). The filtrate was combined with the wash and poured into acetone (1 with stirring. The resulting precipitate was collected by filtration to give 9.52 g of yellow-colored crude product. To a suspension of the crude material (9.5 g) in water ml) was added a small amount of charcoal, and the mixture was filtered. The filtrate was added dropwise, with stirring, to isopropyl alcohol (700 ml). The resulting precipitate was i cc After drying over MgSO 4 removal of the solvent afforded 3.3 g of yellow oil. A solution of the oil in CH 2 Cl 2 (50 ml) was filtered 117 collected by filtration, washed with a small amount of methanol ml), and dried to give 7.58 g of the title compound XXII-H (E isomer) as the hydrochloride. Light yellow powder. Estimated purity 85% by UV. M.p. 173-188 0 C (dec.).

IR KBr cm- 1 1795, 1680, 1620, 1575, 1540.

max UV Phosphate buffer (pH 7) n (E 1 294 (457).

max 1 cm N RR 6D 2 0+DC ppm 3.82 (2H, 5.17 (1H, d, J=5 Hz), 5.33 (2H, d, J=7 Hz), 5.43 (1H, d, 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 8z), 9.00 (2H, d, J=7 Hz).

Preparation No. 19 2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetyl Chloride Hydrochloride (III-1 as its acid chloride hydrochloride) i A. 2-Cyano-2-methoxyiminoacetamide To a stirred mixture of a-cyanoacetamide (252 g, 3 mole) and sodium nitrite (414 g, 6 mole) in water (600 ml) was added acetic acid (371 ml, 10 mole) at 5-10 0 C over 1.5 hours.

The 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 sulate (568 ml, 6 mole) at 15-20 0 C and the mixture was stirred at 45°C for 1.5 hours. The reaction mixture was adjusted to pH 8.5 with 6 N NaOH and allowed to stand at 5°C overnight to separate the precipitate, which was collected by filtration, washed with cold water and air-dried to give 292 g of the title compound as brown needles melting at 170-172°C.

S. Trippett, D. M. Walker, J. Chem. Soc. 19 61 1266.

IR V KBr m1 max 3400, 3180, 1720(sh), 1715, 1690, 1615, 1570.

UIV X 1 2 0 nm 238.5 (8290), 268 (sh, 3870).

I'ma x NMR: 6 DMSO-d 6 4.20 (3H, s, OCH 3 )f 7.85 (2H, br.

PPM

NH 2 Anal. Calc'd, for C 4

H-

5 N 3 0 2 Found: C, 37.80; H, 3.97; N, 33 .06 C, 37.43; H, 3,75; N, 32.51, B. 2-Methoxyiminopropanedinitrile A stirred Mixture of 2-cyano-2-methoxyiininoacetamide (88.9 g, 0.7 mole), sudium chloride (70 g) and phosphoxus oxychloride (97 ml, 1.05 mole) in dry 1,2-'Uchloroethane (350 ml) was refluxed for 16 hours. The -srlubles were filtered off through a dicalite pad and Washed dichloroethane. The filtrate and the wash were combined, and poured into stirred ice-water (1.5 L) to decompose the excess of phosphorus oXYchlorido. The organic phase was washed with 1.0% NaHCO 3 500 mI Y water (500 ml x 3) and a saturated NaCJ. solution (500 ml), and dried over MgSO 4 The filtrate was distilled under diminished pressure to give 61,5 g of the title compound boiling at 62 0 C/24 mm Hg. (Lit,, b.p. 47-.48-C/12 mm Hg), IR V Liquid Film cm& 1 3020, 2960, 22V5, 2020, 1530, 1455, max 1080.

N 11R: 6 CDCl 3 PPM 4,35 O3H, s, 0CH 3 for 2 hours. The clear solution was poured into n-hexane (500 ml) to give a precipitate. The organic layer was discarded by 119 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 2-methoxyiminopropanedinitrile (58.0 g, 0.53 mole) in ethanol (120 ml) at -15 to -10*C over a period of minutes, with stirring. The mixture was stirred at -10 0

C

overnight and then at ambient temperature (20-25 0 C) for one day.

The reaction mixture was partitioned between water (350 ml) and

CH

2 Cl 2 (350 ml), and the aqueous phase was saturated with sodium chloride, and extracted again with CH 2 Cl2 (300 ml). The organic extracts were combined, dried over MgSO 4 and evaporated in vacuo.

A solution of the residue in ethyl acetate (1.6 L) 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. Yield 67.6 g M.p. 152-4°C [Lit,, in.p. 150-155 0 C IR KBr cm-1 3160, 2900, 2360, 2235, 2000, 1665, 1555, max 1495, 1415.

UV 1 EtOH nm 243 (8500), 265 (sh, 5380), 305 (sh, 1400).

max NMR: 6 DMSO-d 6 ppm 1,88 (3H, s, CH 3 COOH), 4.15 (3H OCH 3 7.60 (4H, br.).

Anal. Calc'd for C4H 6

N

4

O-CH

3

COOH:

Found: C, 38.71; H, 5.41; N, 30.09 C, 38.71; 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 CH 3 OH (1.25 L) were added dropwise triethylamine (234 ml, 1.68 mole) at -10 0 C, and subsequently Br 2 S120 (41.6 ml, 0.806 mole) over 20 minutes at -15 to -10 0 C, and the the ti mixture was sti red for 20 minutes. To the mixture was added [Lit.* dropwise a solution of KSCN (78.3 g, 0.806 mole) in CH 3 OH (550 ml) over 1 hour at -15 to -10°C. After stirring at 0-5 0 C for 1 IR hour, the 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 g of the title UV compound. M.p. 263-5 0 C The m.p. of the compound prepared by us is higher by about 60 0 C than that given in the literature* 210-150C but our spectral and F. 2 microanalytical data are consistent for the structure. C KBr IR v K cm 1 3435, 3260, 3120, 2960, 2245, 2020, 1630, max 1545, 1455, 1415. methox (400 IT UV X Et O H nm 248 (13300), 310 (3470). -50 0

C.

max poured

L).

NMR: 6 DMSO-d 6 with t ppm 4.21 (3H, s, OCH), 8.30 (2H, br. NH 2 pressu Anal. Calc'd for CsH 5

N

5 OS: C, 32.78; H, 2.75; N, 38.23: S, 17.50 IR Found: C, 32.76; H, 2.51; N, 38.02; S, 17.50.

*Japar E. 2-(5-Amino-l,2,4-thiadiazo2-3-yl)-2-methoxyiminoacetic Acid (Brit.

II-1) A mixture of 2-(5-amino-l,2,4-thiadiazol-3-yl-2methoxyiminoacetonitrile (18.3 g, 0.1 mole) in 4 N NaOH (250 ml) was heated at 50-55°C with stirring for 3 hours, The reaction mixture was adjusted to pH 1 with H 3

PO

4 and washed with ethyl acetate (100 ml), saturated with NaCl, and extracted th~:;e times with a mixture of ethyl acetate and tetrahydrofuran (3 1, 300 ml x 2, and 200 ml x The extracts were combined, dried over MgSO 4 and concentrated under reduced pressure. The residue was triturated with isopropyl ether to afford pale yellow crystals of 121 the title acid. Yield 16.8 g M.p. 184-5*C (dec.).

m.p. 1,80-182 0 C IR v r cm- 3460, 3260, 3140, 1725, 1620, 1605, 1545.

max UV X X2 0 nm 234 (13200), 288 (sh, 3620).

max F. 2-CS-Amino- l,2,4-thiadiazol-3-yl)-2-methoxyimiloacetyl Chloride Hydrochlor'le To a suspension of 2-(5-amino-1,2,4-thiadiazol-3-yl)-2,metho~yi~iinoacetic acid (40.4 g, 0.2 mole) in dry CH 2 C1 2 (400 ml) was added PC1 5 (41.6 g, 0.2 mole) in one portion at 0 C. The mixture was stirred for 4 hours at 20 to :-5 0 C, r.

poured into a mixture of n-heptane and isopropyl. ether (2 2 The yellow precipitate was collected by filtratione washed with the same solvent mixture, and dried with KOH under reduced pressure to give 46.0 g of the title acid chloride.

IR V Nu jol cm- 1 1775.

max *Japan Kokai 57-158769 published September 30f 1.982, to Fujisawa (Brit, appl., 3/6/81) A Benzhydryl 7-Ben zyl- id ene amino- 3-tr iphenylphosphon i. ihvl-3cephem-4-carboxylate chloride (XV) 122 Prepaation No.

S

N-,rC CONH H2 N 5> N CH=CH-CH 2 -C1 C 2

H

5 COOCH(Ph) 2 VIII-2 Diphenylmethyl 7-f 2-(5-Amino-,2,4-thiadiazo.3-yl)-2-(Z)ethoxyiminoacet,nido] [3-chloro-1-propenyl J'-3cephep3-4carboxylate (VIII-2, Z isomer) To a mixture of N,O-bis(trimethylsilyl)acetamide (2.3 ml, 9 mm~oles) arid crystalline diphenyilmethyl 7-amino-3-[3- -h lor o-I- -Propen-l1-yl 1-3-ceph em-4-ca rboxyl ate hydrochloride 1:XVIII) (1.338 g, 2.8 mmoles) (from Preparation No. 12.) inmethylene chloride (10 ml) was added 2-(5-amino-l,2,4thiadiazol-3-y1)-24-(Z)-ethoxyiminoacetyI chloride hydrochloride (800 mg, 2.95 mxnoles) portionwise, with stirring, at -10*C and the mixture was allo, to stand at 0 0 C for 2 hours. The 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 VIII-2 as an amorphous powder. Yield 1.70 g Mp. >150 0 C (dec,).

I R v max (KBr) in cm1 3300, 1780, 1720, 1690, 1.380, 1220.

UV Xmax (C 2

H

5 OH) in nm 285 (11000).

14MR :6 (DMSO-d) in ppm 1.26 (3H, t, J=7Hz, CH 2 CH 3 4.25 (2H, q, J=7Hz, C

C

HH) 5.0N8 d-d, J=4 8Hze 6.26 (1H, d, J=IlHzl 3-CM), 6.85 (18, s, CHPh 2 9.53 (18, d, J=8Hz, 7-NH).

123 Preparation No. 21

S

-IOl 'N o N CH=CH-CH -1 H2 2 C 2

H

5 COOCH(Ph) 2 IX-2 *A mixture of E and Z isomers Diphenylmethyl 7-(2-(5-Amino-1,2,4-thiadiazol -3-yl)-2-(z)-ethoxyiminoacetamido] 3-iodo-l-propenyl I -3-cephem-4-carboxyl ate IX-2) A mixture of VIII-2 (1.90 g, 3 mmroles) (from Preparation No. 20) and sodium iodide (1.4 g, 9 mxnoles) in acetone (20 ml) was stirred for 10 m~i.nutes at room temperature and then allowed to stand at 5 0 C foi 3 hours. The mix ture was evaporated under reduced pressure, diluted with ethyl acetate (100 ml), washed with 10% sodium thiosulfate and water, and evaporated under reduced pressure. '?rituri~on of the'reqidue with isopropyl ether gave 1.82 g of the title product IX-2 as a light brown amorphous powder.

IR V max KBr) in cm- 3290,r 177I, 1720f 1670, 1530, 13 70 1220.

UV X max (C 2

H

5 OH) in nm (E 1 304 (199).

1 cm :ia-n~c- 124 Preparation No. 22 -CH=N- N S Nsf CH-CH-CH 2

CONH

COOCH(Ph) 2 XXI-H iodide *E Diphenylmethyl 7-Benzylideneamino-3-[(E)-3-(4-carbamoylpy idinio)-l-propenyl]-3-cephem-4-carboxylate (XXI-H iodide) (E isomer) To a chilled solution of the 3-chloropropenylcephem (XVII, Z isomer, 42.8 g, 90 mmoles) (from Preparation No. 16) in dry DMF (80 ml), was added KI (20 g, 120 mmoles) in one portion, and the mixture was stirred at room temperature. The .reaction 1% was monitored by the ratio UV absorption [E (255 nm)/ 1 cm E1% (320 When the ratio became below 1.10 (after 1 cm minutes), the mixture was diluted ;with 800 ml of methylene chloride, treated with active carbon (4 and filtered. The filter cake was washed with 100 ml of CH 2 C1 2 To the combined filtrate and weshings was added isonicotinamide (14.64 and the mixture war 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 600 ml). The residual brown semi-solid was dissolved in CH 2 Cl 2 (100 ml) and the solution was added dropwise to ethyl acetate (3 L) with vigorous stirring. After drying over P 2 0 5 in vacuo, 57.37 g of the quaternized title product XXI-H was obtained as the iodide. Yellow amorphous powder. Mp. 150-155 0 C This product was identical to that obtained by iodination with Nal (Preparation No. 17).

volume. To the concentrate was added n-heptane (200 ml) to give 125 Preparation No. 23 HC1- COOCH (Ph), XVIII

*Z

Diphenylmethyl 7-Amino-3-(3-chloro-l-propenyl)-3-cephem-4carboxylate hydrochloride (Z isomer) (XVIII, Hydrochloride) A 25% solution of chloroacetaldehyde (69 g, 0.22 mmoles) in CHCl 3 was added to a solution of XVI (80 g, 0.11 mole) in CH 2 Cl 2 (1.1 L) containing N,O-bis(trimethylsilyl)acetamide (16.2 ml, 0.06 mole) at -10 0 C in one portion, and the mixture was allowed stand overnight at 5 0 C. The mixture was concentrated to ca. 0.3 L, diluted with a mixed solvent of ethyl acetate and isopropyl ether 0.6 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 The combined filtrate and washing were concentrated to ca. 0.2 L, t.:eated with Girard Reagent T (60 g, 0.26 mole) and 4N HC1 (220 ml), and seeded with a few crystals of XVIII hydrochloride.

After stirring for 3 hours, the resulting crystals were collected by filtration, washed with water (0.5 L) and ethyl acetate L) and dried in vacuo to give 37 g of the title compound XVIII hydrochloride, melting at >185 0 C Pale yellow needles. This product was identical to that obtained in Preparation No. 12.

"rsTfs~-~ 126 Preparation No. 24

S

HC1*

H

2

N

N 0 CH=CH-CH -C1 COOCH(Ph) 2 XVIII *Z Diphenylmethyl 7-Amino-3-(3-chloro-l-propenyl)-3-cephem-4carboxylate hydrochloride (Z isomer) (XVIII, Hydrochloride) To a solution of chloroacetaldehyde (25% solution in

CHCI

3 628 mg, 2 mmoles) in CH 2 Cl 2 (10 ml) were added N,0-bis- (trimethylailyl)acetamide (0.135 ml, 0.5 mmole' and XVI (728 mg, 1 mmole), successively, at 5 0 C. The mixture was allowed to stand overnight at 5 0 C. The mixture was evaporated and diluted with a mixture of ethyl acetate and isopropyl ether 10 ml).

Insolubles were removed by filtration and the filtrate was concentrated to ca. 5 ml. The concentrate was treated with 4N HCI (2 ml), seeded with XVIII hydrochloride and stirred for 1 hour at room temperature. The crystals were collected by filtration, washed with ethyl acetate (10 ml) and water (10 ml) and dried in vacuo to give 384 mg of the title compound XVIII hydrochloride, melting at >185 0 C Pale yellow needles. This product was identical to that obtained by Preparation No. 12.

Preparation No. 2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)acetyl chloride hydrochloride (III-3 as its acid chloride hydrochloride A. Methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)- 2-(propen-3-yloxyimino)acetate A mixture of 685 mg (3.37 mm.oles) of N-(propen-3yloxy)phthalimide [prepared according to the procedure of E.

-I

127 Grochosaki J. Jurczak, Synthesis 1976 682] and 175 mg (3.35 mmoles) of hydrazine hydrate in 5 ml of C 2

H

5 OH was stirred for 1 hour at room temperature. The resulting precipitate was filtered off and the filtrate and washings were combined. To the solution was added 967 mg (3.37 mmoles) of methyl amino-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 and fractions containing the major product were combined and evaporated under reduced pressure. Yield 514 mg Mp. 83-86 0

C.

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

UV max (C 2H5 OH) in nm 223 (9700), 242 (10000).

NMR 6 (CDC1 3 in ppm 1.55 (9H, s, BOC-H), 4.40 (2H, d, J=SHz, O-CH2), 5.21 (2H, m, CH2=CH), 5.90 (1H, m, -CH=CH 2 9.50 (111, br.s,

NH).

B. 2-(5-t-Butoxycarbonylamino-l,2,4-thiadiazol-3-yl)-2- (propen-3-yloxyimino)acetic acidl) A solution of 770 mg (2.3 mmoles) of methyl butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-(propen-3-yloxyimino)acetate and 3.5 ml of 2N NaOH solution (7.0 mmoles) iti ml of CH3OH was refluxed for 30 minutes. The reaction mixture was concentrated in icuo and diluted with 10 ml of ethyl acetate-H 2 0 The water layer wa. separated, acidified to pH 2 with 6N HC1 and extracted with ethyl acetate (10 ml x 2).

The ethyl acetate solution was dried over MgSO 4 and concentrated by a rotary evaporator to afford 596 mg of the title compound. Mp. 134-135 0 C (lit 1 mp. 135-136 0

C.

IR max (Nujol) in cm 1 3150, 1745, 1710, 1550.

procedure was repeated 3 times, and the mixture was monitored by reverse phase tic to confirm that all of the starting 7-amino- 128 UV :X max (C 2 H 5 0OH) in nm (e) 223 (11000), 242 (11300).

N 1R 6 (DMSO-d 6 in ppm 1.55 (9H1, s, BOC-H), 4.77 (2H1, d, O-CH 2 5.22 (2H1, m, CH (1H1, m, CH=CH 2 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)l1) A solution of 570 mg (1.74 mmoles) of carbonylamino-1,2,4-thiadiazol-3-yl)--2-(propen-3-yloxyimino)acetic acid in 6 ml of trifluoroacetic acid was cllowed to stand for 1 hour at ambient temperature. Evaporation followed by trituration with 30 ml of isopropyl ether gave 376 mg of the title compound. Mp. 109 0 C (dec.).

IR Vmax(Nujol) in cm- 3180, 1-110, 1545, 1460.

UV X max (C 2 H 5 OH) in nm 245 (13500).

NMR 6 (DMSO-d 6 in ppm 4.77 (2H1, d, J=5UHz, O-C11 2 5.20 (211, m, CH=C11), 6.0 (1H1, m, CH=CH 2 1) Japan Kokai 57-112396 (7/13/82, Fu-jisawa) Brit. appl, 7935538 (10/12/79).

D. .2-(5-Amino-1,2,4-thiadiazol-3-y (pr open- 3-yloxyimwino) acetyl chloride hydrochloride A soluttLon of 350 mg (1.54 mmoles) of 111-3 and 410 ng (1.97 mmoles) of phosphorous pentachioride in dichioromethane ml) was stirred for 1 hour at 25 0 C. The reaction mixture was poured into 60 ml of n-hexane and the precipitate was filtered off. Yield 323 mg.I IR v ax(Nujol) in 1765.

rnmo1e I[ Li Lly i 11OL LI11IM.Ut k, 4U II.L j a.lIu Lilt! JIIILU Lt: Wdc t~uLj trated under zeduced pressure. The concentrate was allowed to stand at room temperature for 1.5 hours and washed with isopropyl 129 Preparation No. 26 2-(5-Amino-1,2,4-thiadiazol-3-yL)-2-propargyloxyiminoacetyI chloride hydrochloride (111-4 as its acid chloride hydrochloride) A. Methyl 2-(5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl)- 2-propar gyloxyiminoacetate A suspension of 870 mg (4.32 mmoles) of N-propargyloxyphthalimide 1 and 200 mg (4.0 rnxtoles) of hydrazine hydrate in ml of ethanol was stirred at 25*C for 1 hour and filtered. To the combined filtrate and washings was added 1.0 g (3.86 mmoles) of methyl 2-(5-t-butoxycarbonylamino-l,2,4-thiadiazol-3-yl)-2oxoacetate 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 of the title product. Mp. 72-75 0

C.

I :V max (KBr) in cm- 3200, 2380, 1745, 1-710, 1610.

UiV X mx(C 2H5O)in nm 235 (12200).

IIR 6 (DMSO-d 6 in ppm 1.56 (9H, S, BOC-H)( 3.55 (lii, t, J=2Hz, C- CH), 4.85 (2H, d, J-'2Hz, -CH 2 C-CH), 8.9 (18, br.s, NH).

1) Commercially available, Aldrich, 2) 1. Csendes et al., J. Antibiotics 36t 1020 (1983).

2-(5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2propargyloxyiminoa,-etic acid A solution of 490 ma (1.4 mmoles) of methyl butoxycarbonylamino-1, 2,4-tbtiadiazol-3-yl -2-propargyloxyiminoacetate and 2.2 ml of 2N aqueous NaOH solution (4.4 minoles) in 14 ml1 of CH 3 OH was refluxed for 30 minutes. The reaction mixt~ire was concentrated tinder reduced pressure and 10 ml of ethyl ml) was added a small amount of charcoal, and the mixture was filtered. The filtrate was added dropwise, with stirring, to isopropyl alcohol (700 ml). The resulting precipitate was 130 acetate-H 2 0 was added to the solution. The separated water layer was acidified to pH 2 with 6N HC1 and extracted with ethyl acetate (2 x 10 ml). Drying over MgSO 4 followed by evaporation of the organic layer gave 149 mg of the title product. Mp.

135 0 C (dec.).

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

max UV max (C 2

H

5 OH) in nm 233 (11500).

NMR 6 (DMSO-d 6 in ppm 1.55 (9H, s, BOC-H), 3.55 (1H, t, J=2Hz, C-CH), 4.89 (2H, d, J=2Hz, CH2 C-CH), 9.0 (1H, s, NH).

C. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetic acid (III-4)3 A solution of 410 mg (1.26 mmoles) of carbonylamino-1,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetic acid in 5 ml of trifluoroacetic acid was allowed to stand for 1 hour at 25 0 C. Evaporation followed by trituration of the residue with 25 ml of isopropyl ether gave 204 mg of the title compound, Mp. 156-158°C (dec.).

-1 IR max (Nujol) in cm 1 3300, 2480, 1730, 1610.

UV max (C2HsOH) in nm 234 (12000).

NMR 6 (DMSO-d 6 in ppm 3.52 (1H, t, J=2Hz, CSECH), 4.86 (2H, d, J=2Hz, CH 2 -CSCH), 8.10 (2H, br.s, NH 2 1 i 131 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 III-4 and 182 mg (0.88 mmole) of phosphorous pentachloride in dichloromethane (2 ml) was stirred for 1 hour at -5 0 C. The reaction mixture was poured into 30 ml of n-hexane and the precipitate was filtered off. Yield 65 mg IR ma x (Nujol) in cm 1 1770.

Preparation No. 27 2 -(5-Amino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloximinoacetyl chloride hydrochloride (III-5 as its acid chloride hydrochloride) A. Methyl 2-(5-t-butoxycarbonylamino-1,2,4-thi ad iazdiaol-3-yl) -2cycl opentyloxyiminoacetate A suspension of 860 mg (3.7 mmoles) of N-(cyclopentyloxy)phthalimide 1 and 185 mg (3.7 mmoles) of hydrazine hydrate in 5 ml of C 2 H OH was stirred for 1 hour at ambient temperature ani filtered. The filtrate and washings were combined and added to 1.06 g (3.7 mmoles) of methyl butoxycarbonylamino-1,2,4-thi3diazol-3-yl)-2-oXoacetate The solution was allowed 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 evaporation gave the title product. Yield 906 mg Mp. 115-118*C.

IR Vma (KBr) in cm-1 3200, 1745, 1710, 1550.

UV rpax (C 2 HOH) in nm 217 (1800), 252 (7600).

-ll 4' -r I ri 13' NMR 6 (CDC1 3 in ppm 1.51 (9H, s, BOC-H), 1.60 (8H, H<i 3.88 (3H, s, OCH3), 4.90 (1H,

H

br.s, Q 8.70 (la, br.s, NH).

1) U.S. Patent 3,971,778 (7/27/76; Glaxo), Brit. appi. 49255 (10/25/72).

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

B. 2-(5-t-Butoxvcarbonylamino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetic acid A solution of 500 mg (1.34 mmoles) of methyl 2-15-tbutoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetate and 2N NaQH solution (2 ml, 4 mmoles) in 15 ml of

CH

3 OH was refluxed for 30 minutes. The reaction mixture was evaporated and 10 ml of ethyl acetate-H 2 0 was added to the solution. The water layer was separated, acidified to pH 2 with 6N HCl and extracted with ethyl acetate (10 ml x The organic layer was washed with brine, dried over MgSO 4 and concentrated under reduced pressure to give 377 mg of the title compound. Mp. 185 0 C (dec.).

IR VMax (KBr) in cwC 1 3160, 1710, 1550.

UV Xmax (C 2 H50H) in nm 238 (13300).

NMR 6 (DMSO) in ppm 1.51 (9H, s, BOC-H), 1.70 (8H, br.s,, H-C 4.82 (1H, m, X] 0 C. 2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-cyclcpent loxyiminoacetic acid (III-5, z isomer) 3 A solution of 348 mg (0.97 mmoles) of carbonylamino-1,2,4-thiadiazo1-3-yl)-2- yclopentyloxyiminoacetic

I

:i r 133 acid in 2 ml 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 ml of isopropyl ether and 10 ml of hexane to give 215 mg of the title compound. Mp. 162-165 0 C (dec.) (li3): mp. 160-165 0

C

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

max UV max (C 2

H

5 OH) in nm 238 (13300).

NMR 6 (DMSO-d 6 in ppm 1.17-2.10 (8H, 4.60-4.98 (1H, 8.22 (2H, s).

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

D. 2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetyl chloride hydrochloride A solution of 190 mg (0.74 mmole) of III-5 and 2:9 mg mmole) of phosphorous pentachloride in dichloromethane ml) was stirred for 1 hour at room temperature. The reaction mixture was poured into 50 ml of n-hexane. The resulting precipitate was collected by filtration. Yield 122 mg -l IR max (Nujol) in cm 1 1760.

Preparation No. 28 Benzotriazol-l-yl-2-(5-amino-i,2,4-thiadiazol-3-yl)-2-methoxyiminoacetate A mixture of l-hydroxybenzotriazole (2.7 g, 20 mmoles) and dicyclohexylcarbodiimide (4.12 g, 10 mmoles) in 65 ml of DMF was stirred at room temperature. After 15 minutes, III-i (4.04 g, 20 mmoles) was added to the stirring mixture at 0 C, and stirring was continued for 3 hours. The reaction mixture was i c. 134 filtered to remove the insoluble urea, and the filter cake was washed with a small volume of DMF. The filtrate and washings were combined and poured into 800 ml of ice water. The precipitate was collected by filtration to give 5.24 g of the title compound as a light grey powder. Mp. 189-192 0 C (dec.).

-1 IR (KBr) in cm 1815, 1620, 1540, 1415, 1090, 1060, ma x 1005, 945, 865, 740.

UV Xmax (C 2

H

5 0H) in nm (E 1 246 (580), 283sh (228).

1 cm

Claims (12)

1. A compound of the formula S N CONH R 1 N\ OR 2 h=CII-CH 2 z COOB1 wherein R 1is hydrogen or a conventional amino-protecting group, R .is hydrogen, a straight or branched chain alkyl group contain- ing from 1 to 4 carbon atoms, a cycloalkyl or cycloal)kenyl ring containing from 3 to 6 carbon atoms, or a group of the formula R 4 R 4 COOH 13 1 CH R ,or R R -C-COOHi in which R 3is hydrogen, (lower)alkyi or carboxyl, X is halcae~n hydroxy or Clower)alkoxy, and R and R. are eacine-nd, hydrogen, methyl or ethyl, or R 4 and R 5, taken together wi_, carbon atom to which they are attached, mpay be a cycloalkyi---,_ ring containing from to 5 carbon atoms, B 1 is hydrogen or a conventional carboxyl-protectinq group and Z is chloro, bromc iodQ); or a salt, hydrate, solvate or ester thereof.

2. A compound of Claim 1 wherein Z is chloro or iodo, a. IR 2 is~ (lower)alkyl, cycloalkyl of 3 to 5 carbon atoms, 1-carboxycycloalk-l-yl of 3 to 5 carbon atoms, alkyl, propar.yl or carboxy(lower)alkyl; or a salt, hydrate, solvate or ester t her ecf, -135-

3. A compound of claim 2 wherein R 2 is methyl, ethyl, cyclopentyl, allyl, or propargyl.

4. A compound of claim 3, wherein BI is hydrogen or a benzhydryl group and RI is hydrogen or a trityl group. The compound of claim 4 which is diphenylmethyl

7-[2-(5-amlno-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacet- amido]-3-(3-iodo-l-propen-1Vyl).3-cephem-4-carboxylate, or a salt, hydrate, solvate or ester thereof. 6. The compound of claim 4 which is diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol-3.yl)-2-ethoxyininoacet- amidoj.3-(3-iodo-1.propen-1-yl)-3-cephem-4-carboxylate, or a salt, hydrate, solvate or ester thereof. 7. The compound of claim 4 which is diphenylmethyl 7-[2.(5-amino-1,2,4-thiadiazol-3.yl)-2-cyclopentyloxyimino- acetamido].3-(3-iodo-1-propen-1-yI).3-cephem-4-carboxylate, or a salt, hydrate, solvate or ester thereof.

8. The compound of claim 4 which is diphenylmethyl amidoj.3-(3-iodo-1.propen-1.yl).3-cephem-4-carboxylate, or a salt, hydrate, solvate or ester thereof.

9. The compound of claim 4 which is diphenylmethyl 7-[2-(5-amino-1,2,4-thiadiazol.3-yl)-2-propargylox~yimino- acetamido].3,(3-iodo-1-propen-1.yl),3-cephem-4-carboxylate. or a salt, hydrate, solvate or~ ester thereof. The compound of claim 4 which is diphemylmethyl 7. [2 ano-1,2 thiadiazol.3 -yl) .2 methoxyiminoacet- amido1.3-(3-chloro-1-propen-1.yl).3-cephem.4-carboxylate, or a salt, hydrate, solvate or ester thereof.

11. :The compound of claim 4 which is diphenylmethyl 7-[2-(5.amino-1,2,4-thiadiazol.3.yl).2-ethoxylminoacet- amidoj.3-(3'chloro-l-propen-1-yl).3-cephem.4-carboxylate, or a salt, hydrate, solvate or ester thereof.

12. The compound of claim 4 which is diphenylmethyl 7-[2,(5.arino-1,2,4-thiadiazol.3.yl).2-cyclopentyloxyimino- acetamido]-3-(3-chloro-1.propen-1-yl)-3,cephem.4-

136- carboxylate, or a salt, hydrate,, solvate or ester thereof. 13. The compound of claim 4 which is diphenylmethyl 7-[ 2 -(5-amino-1, 2 ,4-thiadiazol-3-yl).2-allyloxyiminoacet amido]-3-(3-chloro-l-propen-1-yl).3-cephem.4-carboxylate, or a salt, hydrate, solvate or ester thereof. 14. The compound of claim 4 which is diphenyl~methyl 7-[2-(5-amino-l, 2 ,4-thiadiazol-3.yl).2-propargyloxyimino acetamido]-3-(3-chloro-1.propen-1-yl).3-cephem.4- carboxylate, or a salt, hydrate, solvate or ester thereof. A compound of the formula H IN T"xii -N CH=CH-CH 2 NQ wherein -N Q is a quaternary ammonio group; or a salt, ester, solvate or hydrate thereof. 16. A compound of claim 15 wherein (D N Q is selected from

137- 16 Ri 14 RI S 1 16- R R 1 7 R 17 -Na8 -N -NH 2n R 18 1 and z wherein R1 3 R 14 and R 15are the same or different and are (lower)alkyl, (Jlower)alkenyl, amino(lower)alkyl with the pro- vision that the amino may not be on an a-carbon, or hydroxy- (lower)alky. with the provision that the hydroxy group may not be on an a-carbon; R 1 6 is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)- alkylthio, amino, (lower)alkylamino, di(lower)alkylamino, form'yi- amino, (lower )alkanoylarrino, carboxy, hydroxy, carboxy(lower)- alkyl, carboxy(lower alylthio, hydroxy(lower )alkyl, .halo(lower)alkyl, amino(lower)alkyl, (lower)alkoxy(lower )alkvl, carbamoyl or N-(lower)alkylcarbamoyl, or R V may represent a divalent alkylene group having 3 to 5 carbon atoms; R 17is (lower)alkyl, (lower)alkoxy(l.ower)a'Lkyl, a (lower)alkylt allyl, hydroxy(lower)alkyl with the provision tht t~Ie hydroxy group is not on the a-carbon, amirio(lower)alkyl the provision that the amino group is not on the a-carbon, phenyl (lower) alkyl; 18 is hydrogen, (lower)alkyl, (lower)alkoxy, (lower>- alkoxy(lower)alkyl, (lower)alkylthio, amino, (lower)alkylam'inc, di(lower)alkylamino, carboxy, hydroxy, carboxy(lower)alkyl, hydroxyllower)alkyl, amino(lower)alkyl; formylamino, (lower)- alkanoylamino, carbamoyl or N-(1ower)alkylcarbamoyl; -138- n is an integer of from I to 3, Inclusive; Z is Cfl 2 or, when n is 2, Z also may be S, 0Oor N.R 1 9 in which R1 9 is hydrogen or (lower)alkyl; and R 2 0 and R 2 1 are the same or different and are hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkylthio, amino, (lower)alkylamino, di(lower)alkylamino, carboxy, hydroxy, hydroxy(lower)alkyl, amino(lower)alkyl, (lower )alkoxy( lower) alkyl carboxy(lower)alkyl, carboxy(lower)alkylamino, (lower)alkanoylamino, carboxy(lower)alkanoylamino, carbamoyl or N-(lower)alkylcarbamoyl; or a salt, ester, solvate or hydrate thereof. 17. A compound of claim 15 wherein N- Q I -methylpyrrolidinio. 18. A compound of claim 15 wherein N -Q is pyridinio, 19, A compound of claim 15 wherein N. Q is 2 amino. 5.th,,.azolo[4, 5. cipyridinio. A compound of claim trimethylammonio. 21. A compound of claim 3-amino. pyridinio. 22. A compound of claim 3-formylaminopyridinio. 23 A compound of claim 3- cat.bamoylpyridlnio. 24. A compound of claim 4-carbamoylpyridinio. 15 wherein N--Q is 15 wherein N--Q 15 wherein N==Q 15 whecein N- Q is 02 15 wherein N- Q is 139 w A compound of claim 3-aminomethylpyridinio. 26. A compound of claim 2-methylthiazolio. 27, A compound of claim 3-hydroxymethylpyridinio. 28. A compound of cldim 4-hydroxymethylpyridinio. 29. A compound of claim 15 wherein N-Q is 15 wherein is 15 whdrein is 15 wherein N Q is G 15 wherein Nz-EQ is 4- (N-me thylcarbamoyl) pyridinio. A compound of claim 15 wherein N. Q 4-carboxypyridinio. 31. A compound of claim 15 wherein N Q 2,3. propylenepyridinio, 32. A compound of claim 15 wherein N -Q 3. carboxymethylpyridinio. 33. A compound of claim 15 wherein NEE _Q 4. carboxyme thyl thiopyridinio. 34. A compound of the formula R 2 3 R' CH= NS MxIX C00R1 2

140- wherein R 22 is hydrogen or a conventional carboxyl-protecting group, and R 2 1, R 24 and R 25 are the same or different and are hydrogen, hydroxy, (lower)alkyl or (lower)alkoxy and z is chioro, bromo or lodo; or a salt, solvate, hydrate, or ester thereof. A compound of claim 34 wherein R 22 is the benzhydryl group and z is chloro or iodo. 36. A compound of claim 35 wherein R 2 3 R 2 4 and R 25 are hydrogen. 37. The compond of claim 36 which is benzhydryl 7-benzylideneamino-3-(3-ch.lool-propen-1-yl].3-cephem-4- carboxylate. 38. 'The compond of claim 36 which is benzhydryl 7-benzylideneamino.3.[3-iodo-1-propen-1.yl]-3-cephem.4- carboxylate, 1.41. 39. A pxocess for the preparation of compounds of the formula VIII R 23 R2 4J =N S VIII 25 N ICC 2 R OO 2oCC wherein R 2is hydrogen or a conventional carboxyl-protectiig group, and R 23 R 24and R 25are the same or d 'ifferent and are hydrogen, hydroxy, (lower)al1kyl or (lower)alIkoxy and Z is chioro, bromo or jodo; or a salt, solvate, hydrate, or ester thereof, which comprises, reacting a compound of formula II, R 2 3 R 24 /ClioI 2 wherein R 23 1 R 24 and R 25 are as defined above, with a compound of formula III# -142- H2SN CH 2 C1 0 COOR 2 wherein R 2is a conventional carboxyl-protecting group, to give a compound of formula IV, R 2 3 07: CH 1C1 00R 22 then reActing .ompound of formula IV with sodium iodide or potassium icl.-e to give a coripound of formula 'I R 23 P.4 /L 4 -HS V R then reacting compound of formula V with trlphenylphosphino to give a compound of formula V!, -143- 1iQ A 4 1P (Ph )3 or reacting a compound of formnula IV with triphenyiphosphine to give a, compound of formula VI, then reacting a compound of formula VI with a base to give a compound of formula VII, (Ph) 3 00R2 then reacting a compound of formula VII, with ZCH 2 CHO, whe--in 7 is, chlotine 'bromine or iodine, to give a compound of formnula 23 R24 CH=N S N CH=CH-CH z -14 4- -sj /AIUILt.j± OL Z-kz-t-UtOXy- carbonylamino-1,2,4-tiadiazol-3-yl) -2...oe.ycloenyimiox tic 4 and finally if desired removing by conventional means the carboxyl-protecting group R 22 to give the corresponding free acid and converting said free acid to its salt or ester thereof. A process for the preparation of compounds of the formula R 1 U S OR 2 wherein R 1 is hydrogen or a conventional amino-protecting group, 2 R is hydrogen, a straight or branched chain alkyl group con- taining from 1 to 4 carbon atoms, a cycloalkyl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula R 4 -CH=CH-R 3 R R 4 CC-R3 R R 4 -C-COOH R in which R 3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, 4 5 hydroxy or (lower)alkoxy, and R and R are each independently hydrogen, methyl or ethyl, or R 4 and R 5 taken together with the carbon atom to which they are attached, may be a cyclo- alkylidene ring containing from 3 to 5 carbon atoms, B is -145- stirring was continued for 3 hours. The reaction mixture was hydrogen or a conventional carboxyl-protecting group and Z is chloro, bromo or iodo, or a salt, hydrate, solvate or ester thereof, which comprises reacting a compound of formula I, H 2 N S NN CH 2 C1 COOB 1 with a compound of formula II N -T -COOH II wherein R R and B are as defined above, to give a compound of formula III, N c- CONH I I I R 1 HN S 0 C CH 2 C1 COOB 1 L .4 then reacting compound of formula III with sodium iodide or potassium iodide to give a compound of formula IV, bS C- CONE RIHN S 0CH 2 1 COOB 1 then reacti~ng compound of formula IV with triphenyiphosphine to give a compound of formula V, R 1 HN0 H~R HP (Ph) 3 COoB 1 or reacting a compound of formula III with triphenylphosphin'e to give a compound of formula V, then reacting compound of formuila v with a base to give a compound of formula VI, S N F .ONH R N T H-p(Ph) I 00081 -147- i g HI-I then reacting compound of formula VI with C1CH 2 CHO to give compound of formula VII, S N 'CONH 2 N Rl S S CH=CHCHC1 VIl COOB 1 then reacting compound of formula VII with sodium iodide or potassium iodide to give compound of formula VIII, or reacting compound of formula VII with sodium bromide or potassium bromide to give compound of formula IX N- C CONH S R S/ 2 N V RH s OR H CH=CHCE 2 00oo81 N CONE O R (H=CHCHr IX COOB 1 and finally removing all blocking groups by conventional means and if desired converting the free acid compound to its salt or ester thereof. -148- ther ecf -13 41. A process for the preparation of compounds of the formula xi H 2N 0x CH=CH.cH 2 -N=-Q wherein -N Q is a quaternary ammonio, group; or a salt, ester, solvate or hydrate thereof, which comprises reacting a compound of formula I CHOI with a compound of formula II -S H 2 N i 0 J_:N CH 2 C1 COOCH (Ph) 2 -149- a* ~L AJ .J jJL j-L X-J-1-J~k fl~ JW -136. to give a compound of formula III, a CH=N S 9 CHzC1 COOCH (Ph) 2 then reacting a compound of formula III with sodium iodide or potassium iodide to give a compound of formula IV, S 0 CH= N N 0 COOCH (Ph) 2 then reacting compound of formula IV with triphenyiphosphine or reacting compound of formula III with triphenyiphosphine to gi;- compound of formula V, S e CH=N N y N H P (Ph) 3 COOCH (Ph 2 -150- -137. then reacting compound of formula V with a base to give compound of formula VI, S N CH=P(Ph) 3 COOCH(Ph) 2 further reacting compound of formula VI with C1CH 2 CHO to give compound of formula VII, CH=CH CH 2 C1 COOCH(Ph) 2 then reacting compound of formula VII with sodium iodide or potassium iodide to give compound Of formula VIII, CH=N VIII COOCH(Ph) 2 then further reacting compound of formula VIII, with a secondary -151- alkanoylanino, carbamoyL or N-(lower)alky I lcarbamoyl; alkanoylamino, carbamoyl or N-(lower)alkylcarbamoyl; -138- lr II L rl 1. I_ amine N ,R wherein R, RI are each methyl or together form Rpyrrolidine to give a compound of the formula IX, pyrrolidine to give a compound of the formula IXI Q CCOCH (Ph)2 then reacting compound IX with R"Y, wherein R" is a tertiaryamine QDN and Y is chloro, bromo or iodo, to give a compound of the- formula X, H S H=CHCH 2 iQ COOCH(Ph)2 or reacting compound VIII with a tertiary amine QB=N, wherein t. as defined above to give compound X, then treating compound X with Girard Reagent T or HCl to give compound XI, and if desired converting the free acid to a salt, ester, solvate or hydrate thereof. DATED: 3 November 1988 PHILLIPS ORMONDE FITZPATRICK Patent Attorneys for: BRISTOL-MYERS COMPANY J, .5- -152- i