IE58408B1

IE58408B1 - Chemical compounds

Info

Publication number: IE58408B1
Application number: IE86685A
Authority: IE
Original assignee: Bristol Myers Squibb Co
Priority date: 1984-04-09
Filing date: 1985-04-04
Publication date: 1993-09-22
Also published as: SE470260B; FI84830C; GB8721347D0; SE8901225D0; BE902148A; IE58403B1; ZA852236B; IT1190353B; DE3512225A1; NL192925B; YU118987A; PT80246A; SE8501680D0; YU46213B; NL192925C; DD249024A5; RU2056425C1; JPH0350754B2; HU193750B; NZ211659A

Abstract

7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(substituted)-iminoacetamido]-3 -[3-(quaternaryammonio)-1 - propen-1-yl]-3-cephem-4-carboxylates of the formula in which R<1> and R<2> are as defined herein and is a quaternary ammonio group as defined herein, and salts, solvates, hydrates and esters thereof, are potent antibacterial agents. Processes for their preparation and intermediates in such processes are described.

Description

This application relates to 7-l2-(5-amino-l„2,4-thiadiazol-3yl)-2-(substituted)iminoacetamido]-3-[3-(quaternaryammonio)-lpropen-l-yl]-3-cephem-4"Carboxyistes of the formula in which and R are as defined herein and is a quaternary anunonio group as defined herein, and to salts and esters thereof. This invention also relates to processes for the preparation of the compounds of Formula I and pharmaceutical compositions containing at least one compound of Formula I. • Μ A ) U.S. Ter a3 i et f ormula Patent 4., 390,534, issued June 28, 1983 to Tsutomu a 1., discloses cephem and cephem compounds of the wherein RA is amino or protected amino; R" is hydrogen, acyl optionally substituted aryl, substituted alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkenyl oc an 0- oc S-containing 5-membered heterocyclic ring substituted with oxo acylthioalkyl, optionally substituted pyr idimoalkyl, optionally substituted heterocyclylthioalkyl, alkyl, halogen, hydroxy or optionally substituted thiazolioalkyl; and R^ is carboxy or -4 protected carboxy; provided that R is COO when R' is optionally substituted pyridinioalkvl or optionally substituted thiazolioalkyl; and the dashed line indicates either a single or double bond.

European Patent Application No. 13,762, published August 6, 19SO is concordant thereto and has a similar disclosure. (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 d isclosures .

B) European Patent Application Mo. 62,321, published October 13, 1982, discloses cephem compounds of the formula R' COO ι , . „2 .,, wherein R" is amino or protected ammo; R is an optionally substituted lower aliphatic hydrocarbon group, or cycloalkenyl; and the group of the formula \ I is an optionally substituted heterocyclic cation group containing core than one nitrogen atom; and pharmaceutically acceptable salts theieof. Also disclosed are intermediates of the formula 2 wherein R and R are as defined croup and X~ is an acid residue.

C) European Patent Application 1983, discloses cephem compounds above, R is a protected carboxyl No. 74,653, published March 23, of the formula amino or protected amino; wherein R i.

R*' is an optionally substituted lower aliphatic hydrocarbon group, eyelc(1 ower) a1kyl or cyclo (lowerJalkenyl, R^ is (1 ower)a 1kylamino, N-protected (1 ower) a 1kylamino , dι(1 ower)alkylamino, suIfo(lower )alkylamino, hvdroxy(lower ) alkylamino, otected hydroxy(lower ) a 1kylami no , acyloxy(lower)alkyl, (lower )alkoxy(lower )alko xy(1ower)alky1, di ( lower Jalkylamino( lower)alkyl, {1ower)a 1kylfchio(1ower) a1ky1, ( lower )a1kylthio, ( lower ) a1koxy, (lower) a 1koxy(lower ) a 1k oxy, hydr oxy(lower)a 1koxy , acy1(1ower) alky 1, hydroxy(lower )alkyIthio , di (1ower ) alky laminoil ower)a 1kylthiο, N-containing unsaturated 5-membered heterocyclic group, N-containing unsaturated 5-membered heterocyclicthio, or N-containing unsaturated 5 or 6-membered heterocyclicthio ( 1 ower ) a 1ky 1 which may be substituted with suitable substituent(s); and 4 R is hydrogen or (1ower) a 1kyl; or a salt thereof.

D) U.S. Patent 4,332,60 0, isstfed June 1 , 1982 to Tsutomu Teraji et al . , discloses inter alia compounds of the formula 2 wherein R is amino or protected amino; R is (lower ) a 1kyl and X is hydrogen or carbamoyl.

E) European Patent Application No. 47,977, published March 24, 19S2, discloses cephem compounds of the formula (0) T Am —T —C CONH U . N O-R, ch2r1 COO wherein m is 0 or 1; Am is optionally substituted amino; T is a thiadiasolyl moiety (attached to the other groups by two of its carbon atoms); R, is hydrogen, optionally substituted alkyl, cycloalkyl or optionally substitu substituted thiazolio, optionally a 1 kylairmonio or a pyridinio group ed carbamoyl; and R^ is optionally substituted pyrazolio, tri(lower)of the formula in which Ru is substituted (lower)alkyl [the substituent being cycloalkyl, phenyl, hydroxy, alkoxy, halogen, cyano, carbamoyl,_ carboxyl or sulfo), (lower )alkenyl or carboxy-substituted (lower )alkenvl, (lower)alkylthio or carboxy-substituted (lower )alkylthio, amino or monosubstituted amino [the substituent being (lower ) alky 1, (lowerJalkanovl or aminobenzenesulfonyl], di (lower)alkylamino, substituted carbamoyl [the substituent being (lower)alkyl, hydroxy(lower)alkyl, (lower)alkoxy, hydroxy or cyano], di(lower)alkyIcarbamoyl, thiocarbamoyl, cycloalkyl, phenyl, hydroxy, (lower)alkoxy, halogen, (lower )alkoxycarbonvl, (lowerJalkanoyloxy, (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 Ra? and salts thereof.

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

P) European Patent Application Ho. 30,630, published June 24, 1981, discloses 3-vinyl cephem compounds of the formula Ί R CH-CH wherein R is an optionally protected amino-substituted heterocyclic group which may also have halogen, or a group of the formula 7 in which R 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 fi. Λ formula -N^OR'*, wherein R* is hydrogen, cyclo(lower)alkenyl, (lower )alkynyl, (1ower)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 ί £ f ormula COOH in which OR' is methoxy# carboxymethoxy, tert-butoxvcarbonylmethoxy or 1-tert-butoxycarbonylethoxy.

G) U.K. Patent Specification No. 1,399,086 published June 25, 1975, contains a generic disclosure encompassing a vast number of cephalosporins of the formula R-OCO-NE^ORa £ I COOH wherein R is hydrogen or an organic group, R 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 -CH~C in which R and R* independently may be hydrogen, nitrile, (lower)slkoxycarhonyl, or substituted or unsubstituted aliphatic, cycloaliphatic, araliphatic or aromatic. However, the 5-aminoi5 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 anunonio-substituted propenyl group. O.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. ζ' Η ) 14, European Patent Application 1983, discloses compounds of Mo. 88,385, published September th e formula ι ? m which R" is (unsubstituted) thiadiazolyl; R" is carboxy3 (lower)alkyl or protected carboxy (lower )alky1; R is hydrogen, halogen or (lower ) alkenyl; and R is carboxy or protected carboxy Although 1-oropenvl ,3 s listed as one of the possible meanings of 3 R , the application only exemplifies compounds where R is hydrogen, chloro 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 in which R ini ^,R \/l R alia may be alkvl, vinyl, cyanomethyl ,3 OX A protective group such as 2-methoxyprop-2-yl, and R and R' are alkyl groups (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R and R', taken together with the nitrogen to which they are attached, may form a saturated heterocyclic ring of 5 or 6 members, optionally containing another hetero-atom selected from N, 0 and S, and optionally substituted by an alkyl group. The compounds are useful as intermediates in the preparation of 3-thiovinyl cephalosporin derivatives. There is no disclosure or suggestion of a 5-amino1,2,4-thiadiazol-3-yl moiety in place of the 2-sminothiazoi-4~yl substituent or of a quaternary ammonio-substituted propenyl moiety for the 3-substituent. Published United Kingdom Patent Application No. 2,051,062 is concordant thereto and has a similar disclosure.

J) European Patent Application No. 53,537, published June 9, 1982, discloses, inter alia, 3-vinylcephalosporin derivatives of the formula H CH-N coor2 in which R- and r" are the same or different and are hydrogen or alkyl, or taken together, form an alkylene group containing 2 or carbon atoms r| is an acid protecting group, R, is an acid protecting group such as an ester, R^ and R„ are the same or different and are hydrogen, alkyl (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R^ and Rx, taken together with the nitrogen to which they are attached, may form a saturated heterocyclic ring of 5 or 6 members, optionally containing another hetero-atom selected from N, 0 and S, and optionally substituted by an alkyl group.

The compounds are useful as intermediates in the preparation of 3-thiovinyl 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 of a quaternary ammoniosubstituted propenyl group for the 3-substituent.

O.S. 4,423,214 is concordant thereto and has a similar d i sclosu re.

KJ European Patent Application No. 53,074, published June 2, 1982, generically discloses a vast number of 3-vinylcephalosporin derivatives of the formula wherein R' la (in one of sevei embodiments) may be N-AJ .c-coII N OR, in which Rg inter alia may be hydrogen, alkyl, vinyl, cyanomethyl, an oxime-protecting group such as trityl, etc., or a group of the formula C-COOR \b R e in which Ra_ and R^ b hydrogen, alkyl or, carbon atoms, and R R°2S is hydrogen or are the same or different, and may be taken together, an alkylene radical of 2 or 3 g is hydrogen or an acid-protecting radical,· an acid-protecting radical such as methoxymethyl R° (in one of several embodiments) may be a methyl group substituted by a 5- or 6-membered aromatic heterocyclic ring containing a single hereto atom, such as 2- or 3-pyridyl, 2- or 3-thienyl or 2- or 3-furyl; and 3 R^ is ® group of the formula R,SO,O4 2 , in which R^ may he alkyl, trihalomethyl or optionally substituted phenyl. i I These compounds are stated to he intermediates in the ί preparation of compounds in which the 3-substituent is a group of fhe formula R° I ~CH=C~SR which are stated to have antibacterial activity, Although this patent includes the possibility of R° being a methyl group substituted by an N-containing heterocyclic ring, in both the intermediates and final products (thus giving a heterocyclic substituted propenyl moiety), it teaches only that the heterocyclic ring is attached via one of its carbon atoms. Thus, there is no suggestion of a quaternary ammonio-substituted propenyl group.

The reference exemplifies R* in the intermediates and final : products only as methyl. Further, in both the intermediates and final product, the propenyl group must contain a second substituent 4 (-0^SR 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.

L) European Patent Application No. 53,538, published June 9, 1982, discloses, inter alia, 3-vinylcephalosporin intermediates of the formula in which n is 0 or 1, R is hydrogen, alkyl, vinyl, cyanomethyl 3 or an oxime-protecting group, and R is halogen. There is no disclosure or suggestion of a 5-amino-l,2,4-thiaciazol-3-yl moiety in place of the 2-aminothia2ol-4~yl subsoituent, and no disclosure or suggestion of a 3-halo-l-propen-l-yl substituent the 3-position.

Complete Disclosure This application relates to novel cephalosporin derive tives which are potent antibacterial agents. More particularly, it relates to comoounds of the formula R^KM' Nn •C li N \ ·) OR CONH CH-CH"CH9-N^Q cocP wherein κ is hydrogen or a conventional amino-protecting group, R is hydrogen, a straight or ing from 1 to 4 carbon atoms, containing from ί branched chain alkyl group contain a cycloalkvl or cycloalkenyl ring o 6 carbon atoms, or a group of the formula ,3 » f or - C-COOH A5 «η in which R is hydrogen, hydroxy or (lowerJalkoxy, hydrogenj methyl or ethyl carbon atom to which they lowerJalkyl or and R4 and R5 or R4 and R5* are attached. carboxyl, X is halogen, are each independently taken together with the may be a cvcloaXkylidene ring containing from 3 to carbon atoms, and is a quaternary ammonio group, end nontoxic pharmaceutically acceptable salts and physiologically hydrolyzable esters thereof Also included within the scope of the invention are the solvates and hydrates of the compounds of Formula 1, as well as the tautomeric forms of the compounds of Formula ι, e.g. the 2-iminothiazolin~4-yl form of the 2-aminothiazol-4-vl moiety. pr ocess In another aspect, this application relates to a >τ the ^reparation of the compounds of Formula I.

As shown in the structural formula, the compounds of Formula I have the ® syn® or " Z ® configuration with respect to the alkoxyimino group. Because the compounds are geometric isomers, some of the 'anti’' isomer may also be present. This invention comprises compounds of Formula I containing at least 90¾ of the "syn" isomer. Preferably the compounds of Formula 1 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 IK) also form geometric (cis and trans) isomers about the double bond of the propenyl group. Both the cis ( Z*) and trans (E") isomers of these compounds are specifically included within the scope of this invention.

The nontoxic pharmaceutically acceptable salts of the compounds of Formula 1 include salts with mineral acids such as hydrochloric, hydrobromic, phosphoric and sulfuric, or with organic carboxylic acids or sulfonic acids such as acetic, trifluoroacetic, citric, formic, maleic, oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic, malic, methanesulfonic, benzenesulfonic, p-toluenesulfonic and other acids known and used in the penicillin and cephalosporin arts. Preparation of these acid addition salts is carried out by conventional techniques. 7 Examples of physiologically hydrolyzable esters of the compounds of Formula I include xndanyl, phthalidyl, methoxypi val oyl oxymethyl , glycyloxymethyl , -methyl-2-oxo-1,3-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. methyl, acetoxymethyl phenylglycyloxvmethyl The compounds of Formula I in which R~ is hydrogen exhibit high antibacterial activity against various Gram-positive 1C and Gram-negative bacteria, and are useful in the treatment of bacterial infections in animals# including man. The compounds of Formula I may be formulated for parenteral use in a conventional manner utilizing known pharmaceutical carriers 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 reconstitution before use, e.g. with sterile, pyrogen-free 2f water. The compounds of Formula I may also be formulated as suppositories utilizing conventional suppository bases such as cocoa butter or other glycerides. The compounds of this invention may, if desired, be administered in combination with other antibiotics such es penicillins or other cephalosporins.

When provided in unit dosage forms the compositions will preferably contain from 50 to 1500 mg of the active ingredient of Formula I. The dosage of the compounds of Formula 1 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 dosage for adult human treatment will usually be in the range of from 500 to 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 750 to 3000 mg per day, in divided doses, normally will be sufficient, although higher daily doses of some 3 of the compounds may be desirable in the case of Pseudoronas infections.

The quaternary ammonio group of the formula N: 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 δ in which 8$, R? an(j may be the same or different and may, for example, be (lower)alkyl or substituted (lower)alkvl in which the substituents are, for example, halogen, amino with the provision that the amino group may not be on an c-carbon, hydroxy with the provision that the hydroxy group may not be on an a-carbon, (lower,alkoxy with the provision that the alkoxy group may not be on an α-carbon, (lower ) alkylthio, (lower ) alky lamino, di dower )alkylamino, carbamoyl, (lower )alkenyl, phenyl (lower )alkyl, phenyl or substituted phenyl (in which the substituents may be, for example, halogen, hydroxy, amino, (lowerJalkylamino, di(lower)alkylamino, acylamino, (lower )alkyl, (lower )alkylthio, (lower )alkoxy ) .

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 9 and the like, Ο may be, for example, hydrogen, halogen, amino, (lower ,alkyl, (lower )alkenyl, (1ower)alkyIthio, carboxy, hydroxy, (lower)alkoxy, (lower )a 1koxy(lower )alkyl, halo(lower)alkyl, hydroxyflower )alkyl, amino(lower ) alkyl, (1ower)alkylamino(lower )alkyl, di(lower ) a 1ky1 ami no (lower ) a 1ky1, {lower )alkylamino, di(lower)alkylamino, carboxy(lower )alkyl, carboxy (lower ) alkylamino, carboxy(lower)alkylthio, carbamoyl, N-(lower)alkylcarbamoyl, formylamino, acylamino, acyloxy, phenyl, pyridyl, amidino, guanidino . Where the structure of the heterocyclic 10 ring permits, R and R* , taken together, may be an alkylene group containing from 3 to 5 carbon atoms, e.g. propylene.

Examples of combined acyclic/cyclic quaternary ammonio groups include, for example, those of the formulae 1 and the like, in which R~ may be, for example, (1ower)alky1, ( lower)alkoxy(lower)alkvl# hydroxy (lower)alkyl with the provision that the hydroxy may not be on an α-carbon, carboxy (lower ) alkyl, ami.no( lower )alkyl with the provision that the amino may not be on an c-carbon, (lower ) alkenyl, halodower ) alky 1, allyl,, and R1 may be, for example, hydrogen, hydroxy, halogen, (lower)alkyl, hydroxydower)alkvl, (lower)alkoxy(lower)alkyl, halo (lower )alkyl, amino(lower ) alky 1, (lower ) alkoxy, (lower)alkylthio, (lower)alkenyl, amino, (lowerJalkylamino, di(Iower)10 alkylamino, acylamino, acyloxy, ’carbamoyl, amidinoilower ) alkyl, phenyl, pyridyl, amidino,. guanidino.

Preferred ouaternary-ammonio groups are those of the formulae Ί 3 Ί η Ί 5 wherein RA , R~* and S* are the same or different and are (lower ) alkyl, (lower ) alkenyl, amino(lower ) alky1 with the provision that the amino may not be on an e-carbon, or hydroxy(lowerJalkyl with the provision that the hydroxy group may not be on an e-carbon,· R1" is hydrogen, (lower)alkyl, (lower)alkoxy, (lower)alkylthio, amino, (lower )alkylamino, di (lower ) alkylamino, formylamino, (lower)alkanoylamino, carboxy, hydroxy, carboxy dower )alkyl, car boxy (lower ) alky lthio, hydr oxy (lower ) al ky 1, halodower)rj ί·** alkyl, amino!lower)alkyl, (lower)alkoxy(lower ) alkyl, carbamoyl or N-(lowerJalkylcarbamoyl, or R* may represent a divalent alkylene group having 3 to 5 carbon atoms? R17 is (lowerJalkyl, (lower)alkoxy(lower)alkyl, halo(lower ) alky 1, allyl, hydroxy(lower )alkvl with the provision that the hydroxy group is not on the α-carbon, amino (lowerJalkyl with the provision that the amino group is not on the e-carbon, or phenyl (lower Jalkyl; R R" is hydrogen, (lower Jalkyl, (lower Jalkoxy, (lower)alkoxy(lowerJalkyl, (1owerJalkylthio, amino, (1ower)alkylamino, di(lowerJalkylamino, carboxy, hydroxy, carboxy(lowerJalkyl, hydroxy(lower )alkyl, aminoilower ) alky 1, formylamino, (lower)aIkanoylamino, carbamoyl or N-(lowerJalkylcarbamoyl; n is an integer of from 1 to 3, inclusive; q Z is CH_ or, when n is 2, Z also may be S, 0 or N-R*-', « Λ M in which R ' is hydrogen or (lowerJalkyl· and 2i R and R " are the same or different and are hydrogen, (lower ) alkyl, (1 ower ) alkoxy, (lower ) alky lthio, amino, (lower)alkylamino, di (lower )alkylamino, carboxy, hydroxy, hydroxy( lower ) alkyl, amino(lower ) alky1, (lower ) alkoxy (lower )alkyl, carboxy (lowerJalkyl, carboxy (lowerJalkylamino, (lowerJalkanoylamino, carboxy(lower )alkanoylamino? carbamoyl or N-(lower )alkylcarbamoyl .

Parfcicularlv ©referred Quaternary ammonio grouos are N-(lowerJalkylpyrrolidinio (and especially N-methylpyrrolidinio) trx(lower )alkylammonio (and especially trimethylammonio) , pyri~ dinio? aminopyrid in io, formylaminopyridinio, carbamoylpyridinio, aminotlower Jalkylpyridinio, carboxypyridinio, hydroxy(lower )a Ikylpyr idinio, N-(lower Jalkylcarbamoylpyridinio, (lower)alkvlenepyridinio, S-methylthiasolio and 2-amino~S-thia2oloI4,5-c lpyridinio. χ η , , 2 values οχ R containing from 3 to 5 carbon the compounds of Formula I, particularly preferred are (lower)alkvl (and especially methyl), cycloalkyl atoms, 1-carboxycycloalk-l-yl containing from 3 to 5 carbon atoms, allyl, propargyl and carboxy(1o«er)alky1 (and especially 2-carboxyprop-2-yl). The most preferred compounds of the invention are ) ) δ) ) ) 1) 7~[2-(5-amino-1,2,4~thiadiazol~3~yl)-2-methoxyiminoacetamido]3-( 3 - (tr imethylam.monio) -1-propen-1-yl J-3-ceph em-4-carboxyl ate, 2) 7-(2-(5-amino-1,2,4-thiadiazal-3-yl)-2-methoxyiminoacetamido33 - I 3- (1-me thy Ipyr r ol idinio)-1-propen-1-ylJ -3-cephem-4-carboxylate, 3) 7-[2-(5-amino-1,2, 4-thiadiazol-3-yl) - 2-methoxyiminoacetamido )3-(3-pyr id inio-1-propen-1-yl]-3-cephem-4-carboxylate, 7-(2-(5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido3 3-(3-(3-aminopyr idinio)-1-propen-1-yl]-3-cephem-4-carboxylate, 7-(2-(5-amino-l,2,4-thia diazol-3-v1)-2-methoxyiminoacetamido]3-(3-( 3-form.ylaminopyridinio)-l-propen-l-yl ]-3-cephem-4ca rboxylate, 7-( 2-( 5-am.ino-l, 2,4-th ia diazol-3-y 1) - 2-me thoxy iminoacet amido J 3-( 3-( 3-aminomethyIpyr id in io) -1-pr open-1-yl 3 -3-cephem-4ca rboxylate , 7-(2-(5-amino-l,2,4-th ia diazol-3-y1)-2-methoxyiminoacetamido 3 3-(3-(3~carbamoyIpyridinio)-1-pxopen-l-vl]-3-cephem-4ca rboxylate , ) 7-( 2- (5-am.ino-l, 2,4-th ia di azol- 3-y 1) - 2-me thoxy iminoacetamido ] 3-(3-(4-carbamoyIpyridinio)-1-propen-l-yl3-cephem-4ca rboxylate , ) 7-(2-(5-amino-l,2,4-thiadiazol-3-y1)-2-methoxyiminoacetamido 3 3-(3-(2-methylthiazolio)-1-propen-l-yl]-3-cephem-4carboxylate, 7-1 2- (5-am.ino-l, 2,4-th ia diazol-3-yl) ~2~me thoxy iminoacetamido ]« 3-I 3-(2-amino-5-thiazolo[4,5-e3pyridinio)-1-propen-l-yl3-3cephem-4-ca rboxylate, 7-[2-(5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido3 3-(3-(4-hydroxymethyIpyridinio)-1-propen-l-ylJ-3-cephem-4ca rboxylate, 12) 7-(2-(5-amino-l,2,4-thia diazol-3-yl)-2-methoxyiminoacetamido3 11) 4 3-(3-(3-hydroxymethylpyndinio)-l-propen-l-yl]- 3-cephem-4ca rboxylate , 13) 7-(2-(5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido)3-(3-(4-{N-methylcarbamoyl)pyridinio )-1-propen-1-yl)-3cephem-4-carboxylate, 14) 7-(2-(5-amino~l,2,4-thiadia2ol~3-yl)-2-meth.oxyiminoacetamido]3-(3-(2,3-propylenepyr idini o)-l-propen-l~yl]- 3-cephem-4ca rboxylate, ) 7-(2-(5-amino-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetarn, ido)3-(3-(4-ca rbamoylpyr idinio)-ϊ-pr open-1-yl)-3-cephem-4carboxylate, 16) 7-(2-(5-amino-1,2,4-th iadiazol-3-yl)-2-cvclopen tyloxyiminoacetam.ido}~3-[3-( 4-car bamoy Ipvr id ini o) -1-propen-l-vl )-3cephem-4-carboxylate, ) 7-( 2-(5-am in o-l,2,4-th. iadi a zol-3-yl)-2-allyloxyimi noace t amido )-3-[3-(4~carbamoyIpvridinio)-l-propen-l~yl)-3cephem-4-ca rboxylate, IS) 7-(2-(5-amino-l,2,4-thiadiazol-3-yl )-2-propar gyloxyiminoacetamido)-3-I 3-(4-carbamoyIpyridinio)-l~propen-l-yl)-3cephem-4-ca rboxylate, ) 7- ( 2- (5-am. in o-l, 2,4~thiadiazol-3-yl)-2-me thoxy imi no a cet amido) 3-(3-(4-carboxypyridinio)-l-propen-1-yl )-3-cephem-4ca rboxylate , ) 7-( 2-(5-amin o-1,2,4-th ia diazol-3-yl)-2-ethoxyiminoacetarnido) 3-(3-(4-carboxypyridinio)-2-propen-1-yl)- 3-cephem-4carboxvlate, 21) 7-(2-(5-amino-l,2,4-th iadiazol-3-yl)-2-methoxyiminoacetamido) 3-(3-(3-carboxymethyipyridinioj-l-propen-i-yl]-3-cephem-4carboxylate and 22) 7-(2-(5-amino-l,2,4-th iadiazol-3-yl)-2-me thoxyiminoacetamido) 3-(3-(4-carboxymethyIthiopyr idinio)-1-propen-1-yl)-3--cephem-4carboxyl ate .

The numbering system utilized herein for the various reactants, intermediates and final products is as follows: η» [Roman Numeral] Arabic Numeral | (if appropriate) | Letter aooroor i at e) The Roman Numeral designates whether the compound is a final product (1] or an intermediate or other reactant [all other Roma Numerals). The Arabic Numerals and Letters are not used in thos instances where the overall class (genus) of compounds is meant.

The Arabic Numeral designates the particular meaning o 7 substituent R . 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 indicat this fact. No prime is used if the carboxyl group is unprotected. A ? · 'prime" also is used with the generic R substituent (i.e. R" ) when generically referring to an R" group containing protected carboxyl group.

The Letter at the end of the compound number refers to he particular meaning of the quaternary ammonio group -N=Q .

For convenience, the Arabic Numerals and Letters 9 assigned to some of the preferred R groups and quaternary ammonio groups are set forth below.

Arabic Numeral methyl = ethyl « allyl or ορ a rgy1 cvclopentyl G Lett er A B C D E F G H Ύ J K L M N P Q © « 1-methyl pyr r ol id in io « pyridinio « 2-amino-5-thiazolo(4,5-c]pyridinio « trimethylammonχo = 3-aminopvridinio » 3-formylaminopyridinio « 3-carbamovlpyridinio « 4-carbamoylpyridinio • 3-aminomethylpyridinio « 2-methylthiazolio « 3-hvdroxymethylpyridinio = 4-hydtoxymethyIpyridinio » 4-(N-methylcarbamovl)pvridinio « 4-carboxypyridinio = 2,3-propylenepyridinio * 3-carboxvmethylpyridinio ! = 4-carboxymethylthiopyridinio 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 organisms in six groups. The geometric means of the MIC's determined in these tests are shown in Table 1. (Γ§) ry Table 1 Conf iq Geometric Mean of MIC (mcg/ml) Cmpd. MO. double j ii bond j j (Gp)~I a (5) (G+)-Ib (5) (G-)-Ia (5) (G-)-Ib (G-)-IX (G-)-III (7) (5) (5) I-1A £/2=1/1 0.26 II 0.70 j • 0.05 0.15 0.23 2.4 1-1A E/Z=7/l | 0.13 0. 35 0.029 0.05 0.17 1.4 I-1B _ 1 e* 0.20 0.4 0 0.016 0.0 4 4 1 0.11 1.6 I-IB £/2=1/4 ] 0.35 0.80 i 0.05 0.11 0.35 3.5 I-1C £ 0.10 0.20 0.0071 0.033 0.087 3.8 I-1D £/2=1/1 0.61 1.4 0.10 0.26 0.46 0 λ j' c§ I-1D £/2=10/1 0.30 0.53 0.05 0.076 0.26 1.-3 I-1E E 0.20 0.40 0.0094 0.029 0.10 1.4 ! I-1F E 0.15 0.40 j 0.0094 0.033 0.099 1.2 I-1G 1 E 0.20 0.35 | 0.0094 0.033 'o.io 1.4 I-1H E 0.20 0.40 CL013 0.04 3 0.10 0.97 I-1I p 0.80 1.6 0.10 0.20 0.69 3.1 I-1J I E 0.17 0.35 0.025 0.07 6 0.15 1.6 I-1K E 0.35 0.80 0.02 9 0 « 0 4 4 0.20 3e5 T — i l £ 0.26 0.61 0.029 | 0.088 0.15 T 2. © Ϊ-1Μ P 0. 35 3 0.70 0.029 j 0.10 0.17 2.3 I-1N £/2=7/1 1.2 1.6 0.013 0.066 0.30 5.7 I-1O £ 0.17 0.35 0.029 0.033 0.11 14 I-2H S* 0.20 0 ffi 4 0 0.014 0.057 0.15 1.4 I"2N E 1 1.2 2.1 0.016 0.11 0.35 4.7 I-2N 2 1.4 3.1 0.044 0.15 i III 0.69 10 I-3H E 0.23 0.40 i 0.057 0.10 ι 0.52 1.9 1-4 W P ί & 0.26 0.. 46 1 0.066 0.11 1 0.60 2.6 I»5H 0.13 0.40 0.20 0.46 1 ·? 1 -· * Λ O "SO β a 1-1P E 0.8 1.6 0.013 0.087 5 0.34 1 λ X S! I-1Q E 0.7 I 0.92 0.0095 0.044 i 0.23 , Ϊ d X«s 8 ( G+) -la • Penicillin-sensitive s (G+ ) -Ib • Penicil1in-resistant S_ ( G-) -la Cephalothin-sensitive pneumoniae (1 strain) (G~) -Ib θ Cephalothin-res istant pneumoniae (2 strains) (G-) -II M. morganii (1 strain) Ser. marcescens (2 str ( G-) -III Ps. aeruginosa (7 stra Table 2, below, gives the Protective Dosecn (?D-n) in 50 50 rice for a number of the compounds of formula I against 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. ins ) Table 2 PD50 (mg/kg) ipd. No. S. aureus Smith E. coli Juhl P. aeruginosa A98 4 3 A I-1B 0.44 0.028 7.7 I-IB 0.65 0.072 NT I~1C 0.22 0.013 NT I-1G 0.96 0.021 5.92 I-1H 0.39 0.015 3.9 I-1J 0.35 0.029 NT I-1K 0.53 NT NT I-1M 0.96 NT NT I-1N 2.0 NT NT 1-10 0.26 0.17 NT I-2N 5.0 NT NT NT « Not Tested ι ip Table 3 c *1/2 (min) AUC (mcc hr/ml ) C mpd . No . max (mcg/ml) I-1B 17 21 11 I-1C 21 32 18 I-1D 20 19 11 I-1H 23 16 14 I-1J 19 16 9 . 7 I-1K 24 14 1 4 I-1M 20 23 i 4 I-IN 24 19 16 1-10 28 32 17 I -2N 22 20 12 I-3H 19 4 7 25 I-4H 27 22 16 I-5H 22 32 18 In another aspect, this invention relates to processes for the preparation of the compounds of Formula I. The preferred procedures are shown below in Reaction Schemes la, lb and Ic, while an alternative procedure is shown in Reaction Scheme 2.

The abbreviation Ph represents the phenyl group. Thus, the -CH(Ph)? 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 GOOCH(Ph) N τ-G -COOH \\ Ν ‘V , S·^ OR~ Ψ CONH ( CH,C1 COOCH (Ph) rv c II Η \ 2 OR Ρ(Ph)3 VI base CONH- *5 vii H= P (Ph) 3 COOCH (Ph)., C ICH,CHO CONH «ί i. c· II N \v->2 OR CONKcX •N Ca*C3CH.

OOCH(Ph) X (secondary amine) Q~N V *? pPfe 2.

(‘Cex* V COOCH(Ph) , Ito R R H,N // /\s \\ s OR X &£"V al amin x: «Ί8 ο uldeblock XX ,/\c/ X 2 OR CONE Reaction Scheme la shows two alternate means of going from Compound IX to Compound XII The direct route, utilizing a for the oreoaration of all tertiary amine (XI) is applicable 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 (e.g. pyrrolidine), and this indirect procedure therefore is suitable for the preparation of compounds of Formula I in which the quaternary ammonio group is acyclic or ’mixed* acvclic/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. pycidinio, thiazolio, 2-amino-5-thiazolo[4,5-clpyridinio).

Rea c 11 on Scheme lb II Nal or SI P(Ph) P (Ph) XV cr * \^^ch= p (ph)., I COOCH(Ph)» XVI ·1 C1CH,CHO CH=N· CH"CHCH^C1 0OOCH(Ph) Girard Reagent T or V HCI xv: H,Nη x^-CH-CHC^Cl I COOCH (Ph) « XVIII N —rr—C-COOH ft Ii // Η,Ν~\ OR* as in Scheme la Reaction Scheme lb is a variation of in that the /-amino group of the starting mater protected as a Schiff base during most of the r the desired 7-side chain acid is added later in Otherwise, the general procedure is similar.

Reaction Scheme la ial (II) is eaction steps, and the synthesis. r; R eaction Sc heme 1 c CH»N- CH=CH"CH,C1 xv: COOCH(Ph)2 Mai or KI XIX Q^N XI (ter am H· r!· V 3G N-acylation with III CH=CHCH Θ -N™Q Reaction Scheme lc 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 lc the last step is acylation of the 7-amino group. The relationship between Reaction Schemes la, lb and lc is shown in the following flowchart. 3? VXH x lb ay*· XVI2 Os-GjOi,Cl COCXH(Fh)2 Ouateraisa-cios ©«blocking Coapound I 7-N-Acy1ation x Ic 3 In Reaction Schemes la, lb and lc, the benzhydryl group was shown as the preferred carboxyl-protecting group. It will be appreciated by those skilled in the art that other carboxylprotecting groups, well-known in the art, may be used. The acylating acid III may be used in the form of a derivative such as its acid halide, activated ester, mixed acid anhydride, all of which are well-known in the act. We prefer to utilize it in the form of its acid chloride. Acylating acid III also may have its amino group protected bv any of the common aminoprotecting groups, e.g. N-trityl, N-formyl. The base used to convert the phosphonium iodide (VI or XV) to give the phosphorylide (VII or XVI) may be NaOH, N&jCO-j, IRA-410 (OH-) resin, IRAiCO^ ) resin, or the like, or a mixture tnereor. 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% aqueous solution, a distilled solution (e.g. 70%) or the anhydrous a1dehyde.

We have found that Compound VIII prepared from Compound VII (Scheme la) typically had a Z:E ratio of about 2.Ί at the propenyl double bond. Compound vuj prepared from Compound XVIII (Scheme lb), on the other hand, typically was almost exclusively the 2 isomer. The difference may not be in the route used, but in the conditions utilized in the Wittig reaction (vii to VIII or XVI to XVII). We have also found that the use of an appropriate silyl reagent such as Ν,Ο-bis(trimethylsilylJacetamide 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. Th© reaction is preferably carried out with 2-5 equivalents of the silyl reagent. When the chloropropenyl cephem (VIII) was reacted with Nal in acetone to give the iodopropenyl cephem (IX), the double bond in the propenyl group was isomerized from 2 to E during the iodination. A short reaction oeriod retained the configuration of the parent Compound VIII to a large extent, while a long reaction period gave primarily the £ isomer of Compound IX. However, an excessive reaction time at high temperature gives a lower purity compound 1X. minutes at 25°C and 2 hours at 5°C gives pure IX in good yield. When utilizing Reaction Scheme Ic, we have found that, when iodinating compound XIV with Nal, a purer compound is obtained if the acetone solution is diluted with CC1. when iodination is essentially completed, and the isomerization portion of the reaction is conducted in the acetone-CCl, mixtui έ When iodination of the chloropropenyl cephem (XVII) to the iodopropenyl cephem (XIX) was performed with KI in DMF, the We find that about to E proceeded as fas completed within 45 e XIX without dilution isomerization of the double bond from z the iodination did. The whole reaction minutes at room temperature to give pur with CC1. in the course of the reaction as Compound XII normally was deblocked without purifies15 tion, and the final product (I) was purified by reverse phase column chromatography utilizing a glass column containing the packing removed from, a Waters' Associates PrepPAK~500/C1 θ cartridge. xxi: R‘ c (8 ! I Reaction Scheme 2 \ 2 XOR — CONH' R1-HNZ S -p 1 OR 'Xp^CH^CK-Cn.

. , » COOCH (Ph) , XXIV R~-HN li il H H w g -C!l N h y K XOR2 \ 7 OR~ Q Reduction XXV CONHxs 1© XXVI /f~ ^\^^CH^CH-CH0NSQ COOCH(Ph) i 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 auaternization. Compound XXV is subsequently reduced, and the remainder of Reaction Scheme 2 is as 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 group such as the trityl group.

The acylating acids of Formula 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 L5 and isopropyl. U.S. Patent 4,390,534# referred to in the Prior Art section, above, exemplifies the preparation of a wide variety " o of compounds of Formula III wherein R* is, for example, cyclopentyl, 2-cyclopenten-l-ylf allyl, 2-propynyl, 1-tert. butyloxycarbonyl-1methylethyl, 1-tert. butyloxycarbonyl-l-cyclopentyl, 1-ethoxy2Q carbonyl-l-methylethyl, tert, butyloxycarbonylmethyl, 1-tert. butyloxycarbonyl-2-methylpropyl, tri tyi.

Compound ΙΣ herein (7amino-3-ehloromethyl-3-cephem-4~ -ί carboxylate), used as a starting material in Reaction Schemes 1a, lb and Ic, is a known compound.

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

The present invention also "provides a process for the preparation of compounds of the formula wherein R 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 R ’ ι •C~CH = CH~R' 1 5 R R • 3 -c~c-or COOH R " J-COOH J 5 in which R is hydrogen, hydroxy or (lower)alkoxy, hydrogen, methyl or ethyl (lower)alkyl or carboxyl, X is halogen. ill and R and R"* r 5 and R are each independently taken together with the 3 carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, an d © is a quaternary ammonio group, and nontoxic pharmaceutically 5 acceptable salts and physiologically hydrolyzable esters thereof which process comprises reacting a compound of the formula -> · wherein R" w j CK=CHCH,Z ' 1 COOB* is the same as R or is a group of the formula ,, 1 . . . , B is a conventional are as defined above, ·> grout), 3' is hvdrogen or a conventional in which X, R and R ca rboxyl-protecting amino-protecting group, Z is chloro, bromo or or one, with a tertiary amine Q===N (or secuen m is 1, reducing the sulfoxide by conventional means subsequently removing all blocking groups by convent odo, and m is zer wi th a R’Z) , and, i , and ional means .

The present invention also provides a process for the preparation of compounds of the formula -i wherein R" is hydrogen or a conventional amino-protecting group, 2 R is hydrogen, a straight or branched chain alkyl group contain ing 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 R -C-CH-CK-R3 , 1 5 R •C-C=C-R‘ COOH or C-COOH 1 5 R in which R3 is hydrogen, (lower)alkyl or carboxyl, X is halogen, 4 5 hydroxy or (lower)alkoxy, and R and R are each independently A 5 hydrogen, methyl or ethyl, or R* 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 is a quaternary ammonio group, and nontoxic pharmaceutically 15 acceptable salts and physiologically hydrolyzable esters thereof which process comprises acylating a compound of the formula η7ν· COO XXII with an acid of the formula B*HN Ν' // cII N -COOH \ 2 OR ii: •p s or with, an acylating derivative of said acid, wherein R~ is the sane ,as R" or is a group of the formula X COOB' C—COOB' in which. X, R' and R are carboxyl-protecting group amino-protecting group. as defined above, B* is 2 and B is hydrogen or a a conventional conventional The reactions are carried out in a non-aqueous organic I solvent such as dimethyl sulfoxide, hexamethylphosphoramide, methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile_ or mixtures of such solvents. The reactions are conveniently carried out at a temperature of from -10eC to -t-SO’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, ΧΧΣΙΙ or XXZV; we normally prefer fco utilise from 25% to 100% excess of the tertiary amine.

Carboxyl-protecting groups suitable for use as B in the above reactions are well-known to those skilled in the art and include aralkyl groups such as benzyl, ρ-methoxybenzyl, p-nitrobenzyl and diphenyImethyl (benzhydryl); alkyl groups such as t-butyl; haloalkyl groups such as 2,2,2-trichloroechyl, and other carboxyl protecting groups described in the literature, e.g. in U.K. Patent 1,399,086. w® 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 are also well-known in the art, and include the trityl group and acyl groups such as chloroacetyl, formyl and trichloroethoxvcarbonyl. 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 ~ 1), the 1-oxide is prepared by known procedures such as oxidation with m-chloroperbenzoic acid, peracetic acid, sodium tungstate . 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.

Intermediates useful for the purposes of the present Invention notably include compounds of the formula XXVIII wherein 2 is chloro, bromo, or iodo, R is hydrogen, a straight or branched chain alkyl group co"ntaining from 1 co 4 carbon atoms, a cycloalkvl or cycloalkenyl ring containing from 3 to 6 carbon atoms, or a group of the formula R 1 3 -C-CH=CH~R I Q R n I -C~C=CI 5 R COOH or R I -C-COOH '5 in which R is hydrogen, (lower)alkyl or 4 5 hydroxy or (lower )alkoxy, and R and R 4 5 hydrogen, methyl or ethyl, or R and R , carbon atom to which they are attached, carboxyl, X is halogen, are each independently taken together with the 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.

Intermediates useful f invention notably include or the purposes of the present also compounds of the formula 8 » β A O Vw> ΛΑ XXIX wherein a22 is hydrogen or a conventional carboxyl-protecting group, and R23, R24 and R23 are the same or different and are hydrogen? hydroxy, (lowerJalkyl or (lower)alkoxy; or a salt, solvate, hydrate or ester thereof.

In connection with the intermediates mentioned in the last two preceding paragraphs (i.e. the compounds of the formulae XXVIII and XXIX), we draw attention to our Divisional Application No. Οι *3« Intermediates useful for invention further include, as of the formula the purposes of the will be appreciated present compounds /7 © CHeCH-CH--N5rQ COO wh erein is a quaternary solvate or hydrate thereof. ammonio group; or a sal ester , In connection with the intermediates mentioned in the last preceding paragraph (i.e. the compounds of the formula XXII), we draw attention to our Divisional Application No.

As used herein, the terms acylamino and acyloxy refer to an acylated amino or acylated hydroxy group in which the acyl moiety is (lower,alkanoyl (e.g. formyl, acetyl, propionyl, butyryl, isobutyryl, isovaleryl ), aroyl (e.g. benzoyl), (lower )alkanesulfonyl (e.g. mesyl* ethanesulfonyl), arylsulfonyl (e.g. benzenesulfonyl, tosyl)» or Ο (lower )alky1 , mean straight "(lower ) alkoxy", or branched chain As used herein, the terms (lower )alkylthio" alkyl, alkoxy, alkylthio groups containing from 1 to β carbon atoms, inclusive. Similarly, the terms (lower ) alkenyl and (1ower)alkynyl mean alkenyl or alkynyl groups containing from 2 to β carbon atoms.

In the Examples section which no- followsP procedures are described which either directly illustrate the invention or are of interest in connection with it.

Example 1 7-( 2-( 5-Amino-l ,2,4-thie.diazol-3-yI)-2-methoxyiminoacetamido]-3· ( 3-(l-methylpyrrolidinio)-1-propen-l-yl]-3-cephem-4-carboxylate (I-1A) .10 To a solution of diphenvImethyl 7-(2-(5-amino-1,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(3-iodo-l-propen-1yl)-3-cephem-4-carboxvlate (IX-1) (Z/E=2/l, 150 mg, 0.21 mmole) in ethyl acetate (2 ml) was added a solution of 1-mefchylpyrrolidine (36 mg, 0.42 mmole) in ethyl acetate (I ml) in one portion with stirring. The mixture was stirred for 15 minutes and diluted with isopropyl ether (10 ml) to form a precipitate, which was collected by filtration. A mixture of th solid (130 mg), formic acid (1 ml) and concentrated HCl (0.1 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 Pr epPAK-500/C·, θ cartridge, 100 ml), eluting with water and 10% CH^OH. The desired fractions were collected, and concentrated in vacuo to a small volume and freeze-dried to give comoound (I-1A) (2/E-l/l), melting at mg (12%) of the title >280°C (dec. ).

IR : vKBr cm'1 3400, 1760, 1660, 1610. max UV NMR : ^Phosphate burfer (pH 7) nm (£;1% } 23δ (372), 2θθ { 322 ) max 1 cm _JJ 6D2° 2.31 (4H, m, ϊό ) , 3.12 (3H, s, ppm H m, 2-H £. N j ), 3 .79 (IH, s, 2-H) Λ7- 3=8, CH7N), 4.2 ( 3H, s, OCHg), 5.36 6-H), 5.95 (3H, m, 7 -H & 3~CH=CH), 6 3=10, 3-CH cis) e 7.0 (1 /2H, d, 3=16, Example 2 ocs3 jE.

E-IB 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetarnido]-3[3-pyridinio-l-propen-l-yl]-3-cephem-4-carboxylate (1-13) A mixture of diphenylmethyl 7-(2-(5-amino-l,2,4thiadiazol-3-yi)-2-methoxyiminoacetamido 3-3-(3-iodo-l-propen-lyl)-3-cephem-4-carboxylate (ΣΧ-1) (E, 716 mg, 1 mmole), pyridine (158 mg, 2 mmoles) 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 added to formic acid (6 ml) containing sodium bisulfite (60 mg). The mixture was stirred for 30 minutes at 4 0°C and concentrated to dryness. The residue was dissolved in H9O (40 ml) and some insolubles were removed. The aqueous solution was charged on a column of reverse phase (PrepPAK-SOO/C^g, 100 ml) eluting with H-,0 ( 300 ml) and 5% aqueous CH^OH (800 ml), end the eluate was monitored by uv (254 nm) and HPLC. The fractions (5% aqueous CH^OH) containing the desired product were combined, concentrated to a small volume and lyophilized to yield 40 mg (8%, of the title compound (I-1B), melting at >200°C (dec.).

IR uv K n r - 1 v cm - 3350, 1760, 1660, 1600. ΓΠϋ X ^Phosphate butfer (pH /) nm (£1% } 24Q (352), 258 (366)(maX 1 Cm 267 (279), 290 (469).

HER : ^O+DMSO-dg 3>74 {2H, bc„s> 2-K), 4.20 (3H, s,OCE3), P?rn 5.92 (IH, d, J=4.5, 7-H), 6.15 (IH, m, 3-CH=CH), 7.04 (IH, d, J=16, 3-CH trans), 8.2 (2H, m, Py-H3 $), 8.62 (IH, m, Py-Hj , 8.97 (2H, m, Py-H, g).

Example 3 H2N il 11 Λ N N v OCH.

CONHO // Xz io X-z 1-13 *Z/E=4/j 7-[2-(5-Amino-1,2,4-fchiadiazol-3-yl)-2-methoxyiminoace t.amido)-3[ 3-pyridinio-l-propen-l-Yl]-3-cephem-4-carboxylate (1-13) The chloropropenyl compound, diphenyImethyl 7-(2-(5ami no-1,2,4-thiad iazol-3-yl)-2-methoxviminoacetamido]-3-(3ch ioro-l-propen-l~yl)-3-cephem-4-carboxylate (vui-l) (Z, 937 mg, 1.5 mmoles) was added to a stirred solution of pyridine (237 mg, κ .·ι ·.£' ' ιΉ mmoles) 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 40°C. After removal of the formic acid, the residue was purified by reverse phase column chromatography (packing of PrepPAK-500/C^g cartridge, 100 ml) by eluting with 5% aqueous CKjOH. The fractions containing the desired product were combined on the basis of HPLC analysis, evaporated under reduced pressure and lyophilized to give 41 mg (5.5%) of the title comoound (1-13) (Z/E = 4/l). Mp. >200°C (dec.).

UV NMR KBr -1 υ cm max 3300, 1760, 1660, 1600.

^Phosphate buffer (PH 7) nm (E1% ) 237 (386), 250 (377 ) , max1 Ci? 2 58 (369), 265 ( 34 7 ) , 280 (311). 6D2° 3.45 ε 3.76 (each IH, d, J=16, 2-H), 4.18 (3H, S p ppm OCH3), 5.34 ( 3H , m , CH-CH-CH-, 6 6- H), 5.92 (IH t d e J — 4 .5, 7-H), 6. 58 (4/5H, d, J=ll, 3 - CH cis), 7a03 (1/5H, d, J = 16, 3- CH trans ), 8.12 (2H, m, Py-H 3,5 5 f 8.56 (1 H, m, pyH4 ), 8.82 (2H, m, Py-H^,g)· S 5 - ι 2-(5-Ami no-1,2,4-thiadiazoI-3-yl)-2-methoxyiminoacetamido 3-3[3-(2-amino-5-thi azolo[4,5-c3pyr idinio)-I-or open-l-ylj-3-cephem4-carboxylate (X-IC) A stirred solution of diphenyImethyl 7-[2-(5-amino1,2,4-thiadiazol~3~yl) -2-methoxyiminoacetamido 3-3-(3-iodo-lpr open-l-yl)-3-cephem-4-carboxylate (IK-1) (E isomer, 714 mg, 1 mmole), 2~aminothia2olo[4,5-c)pyridine [prepared according to the procedure of T. Takahashi et al., Pharm. Bull (Japan), 2., 34 (1954)3 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 sodium bisulfite (70 mg) were added to the powder (700 mg), and the mixture was stirred for 30 minutes at 40-45°C. After evaporation, the residue was triturated with H^O (40 ml). Insolubles were filtered off, and the filtrate was chromatographed over a reverse phase column (PrepPAK-500/C1 θ, 100 ml), with H^O and 10% CK-jOH 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 110 mg (19%). Mp. >200°C (dec. ).

IR : vKSr cm1 3300, 1760, 1660, 1630, 1600. max UV : ^Phosphate burfer (pH /) nm (£1% } 2<5 (499)/ 265 (286) max cm N KR &DMSO dg+D20 3g6 (3H, S' ock3), 4.98 (IH, d, J = 4.5, PPm 6-H), 5.2 (2H, m, CH=CH-CH2), 5.57 (IH, m, 3-CH»CH), 5.96 (IH, m, 7-H), 7.16 (IH, d, J-^lSj 3-CH trans), 8.36 & S.45 (each IH, c, J = 7, Py-H), 8.92 (IH, s, Py-H).

Example 5 N· -cII N - CONH H9N OCH- N Ί \^>%‘CH-CH-CH2-N (CH, ) I θ cocH I-1D /£ » 1/1 7-(2-(5-Ami no-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetarnido]-3(3-tr imethylammonio-1-propen-1-yl)-3-cephem-4-carboxylate (I-1D) To a solution of diphenylmethyl 7-(2-(5-amino-l,2,4thiadiazol-3~yl)-2-methoxyiminoacetamido J-3-(3-iodo-l-pr open-1yl )-3-cephem-’4-carboxylafce (IX-l) (Z/E=2/l, 490 mg, 0.68 mmole) in ethyl acetate (14 ml) was added a 0.1 M trimethylamine solution in ether (13.6 ml) in one portion. The mixture was stirred for 10 minutes and evaporated to dryness, and the residue was triturated with ether (20 ml). The resulting solid (490 mg) was added to trifluoroacetic acid (0.2 ml) containing one drop of anisole. After 1.5 hours’ stirring, the mixture was evaporated to dryness under reduced pressure and the residual oil was triturated with ether (20 ml). The resulting precipitate was collected by filtration and dissolved in K?O (20 ml). Some insolubles were removed, and the aqueous solution was eluted on a 7 Ο,θ reverse phase column (the packing of PrepPAK-50O/C^g cartridge, Waters' 30 ml) using water as eluant. Fractions contain ing the desired compound were combined and concentrated to a small volume and lyophilized to afford 30 mg (9.2%) of the title compound (I-1D) (Z/E » 1/1) as a colorless amorphous powder, melting at >15O°C (dec.).

IR : vKBr cm-1 3300, : max 1770* 1670, 1605. UV : ,Phosphate buffer ΛipH z) nm (E1% ) 236 (389), 267 (343) max 1 cm 2θ 3.45 & 3.7 (IH, d, J=16, 2-H), 3.81 (IH, s, 2-H), ppm © 4.1 (2H, d, J=8, -CH-,N), 4.21 (3H, s, OCHg), 5.39 (IH, d, J=4 .5, 6-H), 5.95 (2K, m, 3-CH=CH & 7-H), 6.61 (1/2H, d, J-ll, 3-CH cis) , 7.05 (1/2H, c, J=16 , 3-CH trans).

Example 6 II ***·*%.

Kg —C—CONH II N /θ[2°^·(5" Ajit’ i η ο ~ 1t 2 t ol-3-yl )-2-methoxyiminoacetamido)-3[3-(3- am inopyri din io)-1-propen-1-yl]-3-cephem~4-ca rboxylate ( I -IE) D ipheny Imet hyl 7-[2-(5-amino-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3-(3-iodo-1-propen-1-y1)- 3-cephem-4carboxylate (IX-1) (E, 716 mg, 1 mmole) was added to a stirred solution of 3-aminopyridine (188 mg, 2 mmoles) in DMSO (1 ml). 3 The mixture was stirred for 1 hour and diluted with ethyl acetate (20 ml). The resulting precipitate was collected by filtration, washed with ethyl acetate and dried to give 520 mg of yellow powder. A mixture of the powder (500 mg), formic acid (5 ml) and sodium bisulfite (50 mg) was stirred for 30 minutes at 40°C. The mixture was concentrated in vacuo, dissolved in H_O (40 ml) and filtered to remove insolubles. The aqueous solution was chromatographed on a column of reverse phase (packing of Prep,?AK-500/C^g, 100 ml), with 7.5% aqueous CHgOH elution, fractions containing the desired compound were evaporated and lyophilized to give the title compound (1-1E) (7 mg, 1.4%), melting at >185°C (dec.).

The IR UV NMR KBr -1 υ cm. ma x 3400, 1765, 1675, 1620, 1600 Phosphate buffer (pH 7) max &D2° ppm. nm „1% , Jt ) 1cm 3.72 (2H, m, 2-H), 4.14 (3H, 246 (403), 290 (468) s, OCH ^), 5.35 6-H fe CH=CH-CH2), 5.9 (IH, d, 3=4.5, 7-H), 6.1 (IH, m, 3-CH=CH), 7.05 (IH, d, 3=16, 3-CH, trans), 8.1 (IH, m, Py-Hg), 8.54 (IH, br-s, Py-Kg), 8.68 (IH, m, Py-H^), 9.4 (IH, m, Py-H2).

Treatment of IX-1 (716 mg, 1 mmole) with 3-t~butoxy~ carbonylaminopyridine (324 mg, 2 mmoles) by a procedure simila! to that described above gave 12 mg (2.3%) of I-1E. '3 Example 7 Η,Ν ΪΠΠ -CONBOCHCK \X^CH=CHCH,-N cocP NH.

I-1E 2/E « 1/1 7-(2-(5-Ami no-1,2,4-thiadiazol~3-yl)-2-methoxyininoacetamido]-3[ 3-(3-amino-l-pyridinio)-1-propen-l-y1j-3-cephen-4-carboxylate (I-1E) A mixture of diphenyImethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido ]-3-(3-iodo-1-propen-lyl)-3-cephem-4-carboxylate (IX-1) (Z/E=2/l, 500 mg, 0.7 mmole) and 3-aminopyridine (66 mg, 0.7 mmole) in dimethylsulfoxide (1 ml ) was stirred for 20 minutes at room temperature. The mixture was diluted with ethyl acetate (10 ml) and ether (10 ml), and the resulting precipitate was collected by filtration, washed with ether (10 ml) and dried. The quaternized salt was dissolved m formic acid (3 ml) containing concentrated HCl (0.3 ml) and stirred for 1.5 hours at room temperature. The mixture was concentrated to dryness under reduced pressure. The residue was dissolved in 2% HCl (10 ml) and filtered. The aqueous layer was chromatographed on a reverse phase column (PrepPAK-500/C^g, 100 ml). After washing with water (500 ml), the column was eluted with 5% aqueous CH^QH. The fractions containing the title compound were combined, concentrated in vacuo and freeze-dried to give 15 mg (4.2%) of the title compound (I-1E) (Z/E~l/1) as a colorless amorphous powder. Mp. >160°C (dec.). ΰ IR vKBr cm-1 3400, 1765, 1675, 1620, 1600 max : XPhOSphate buffer (pH 7) nm (£1% } 2^ (434), 2g? max cm NMR : 6DMSO~d6 + D2O 3>73 {2H, 4.14 (3H, 5.35 ( 3?. , Ppm m, 6-H & CH = CH-CH2), 6.0 (2H, m, 7-H 6 3-CH=CH), 6.6 (1/2H, d, J*=ll, 3-CH cis), 7.05 (1/2H, d, J*16, 3-CH trans), 8.08 (IE, m, Py-H^), 8.6 (2H, m, Py~H4 g), 9.4 (IH, Py-H,). h2n N 'U OCH.

CONH cod2> H=CHCH2~ NHCHO I-1F 7-(2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetarnidoj-3[.3-( 3-f or my 1 aminopyr idinio) -l-propen-l-yl ]- 3-cephem~4-carboxy1 at ( I-1P) A mixture of diphenylmethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido)-3-(3-iodo-l-propen-1yl)-3~cephem-4-carboxylate (XX—1) (E, 716 mg, 1 mmole) and 3formylaminopyridine [prepared according to the procedure of N. Enomoto et al., Bull. Chem. Soc. Japan, 4.5, 2665 (1972)] (244 mg 2 mmoles) in DMSO (2 ml) was stirred 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 of the quaternized salt (500 mg) and sodium bisulfite (50 mg) in HCOOH (5 ml) was stirred at 40-50°C for 80 minutes and evaporated to dryness in vacuo. The residue was dissolved in water (40 rai), neutralized with NaHCO^ and then filtered to remove insoluble material. The clear filtrate was chromatographed on a reverse phase column, PrepPAK-50Ο/C., θ (100 ml), with water and 5% CH-jOH, 10% CH3OH, 20% CH3OH and 30% CH3OK The fractions containing the desired compound were combined, concentrated in vacuo and lyophilized to give 16 mg (2.9%) of th title compound (1-1P) (E) as a tan powder. Mp. >170°C (dec.).

IR : vKBr cm'1 3340(br), 1760, 1670, 1620(br), 1590. max uv KKF.

^Phosphate buffer H 7> nm (E1% ) 216 (428), 248 max 1Cm 290 (474).0D,CH-NaHCO 3 3.68 (2H, br, 2-H), 4.15 (3E, s, OCH3), ppm (IH, d, J=4.5, 7-H) , 6.25 (IH, m, CH-CH 6.98 (IH, d, J«16, 3-CH trans), 8.8-7.9 m, Py -H), 9.38 (IH, br, NHCHO).

Example 9 •1G 7-[2-(5-Ami no-1,2,4~thiadiazol-3yl) - 2-methoxyiminoacetamido] -3[ 3- ( 3-carbamoylpyridinio)-l-propen-l-ylJ-3-cephem-4~carboxylate (I-1G) To a solution of diphenylmethyl 7-( 2-( 5-a,mino-l, 2,4thiadiazoI-3-y!)-2-methoxyiminoacetamido)-3-(3-iodo-l-propen-lyl)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) in DMSO ( ml) «as added nicotinamide (244 mg, 2 mmoles), and 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 (50 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, PrepPAK50C/C,, (100 ml). After elution with water and 5%, 10% and 20% aqueous CH^OH, successively, the fractions containing the desired material were combined, concentrated in vacuo and freeze-dried to yield 21 mg (3.8%) of the title compound (I~1G) (E) as a yellow powder. Mp. >175°C (dec.).

IR : vK3r cm-1 max 334C(br), 1760, 1670, 1600. UVλ Phosphate Λ buffer(pH 7) nm (EX% ) 235 (326), 274 (sh, max1 Cm 405), 290 (446). A’ MR : .D,ONaHCO, 0 2 J 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH^, 5.32 PP11'· (IH, d, J-4.5, 6-H), 5.45 (IK, d, J=7, CH-CH -CH?), 5.88 (IH, d, J=4.5, 7-H), 6.15 (IH, d-t, J«16 & 7, 3-CH=CH), 7.00 (IH, d, J-16, 3-CH trans), 8.23 (IH, m, Py-H^), 9.03 ( 2H, m, Py-H4&6), 9.34 (IH, s, Py-H2).

Example 10 (ft fi OCH., Ι-lH CONH 7-(2-( 5-Am.ino -1 ,2,4-thiadiazol-3-yl)2-methoxyiminoacetamido]-3[ 3-(4-carbamovlpyridinio)-1-propen-l-yl]-3-cephem-4-carboxylate (Σ-1Η) To a stirred solution of diphenylmethyl 7-[2-(5-amino1, 2 , 4-thiadiazol-3-yl)-2-methoxyiminoacetarnido J-3-(3-iodo-lpropen-l-yl)-3-cephem-4-carboxylate (IXl) (S, 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°C for 1 hour, and evaporated to dryness under reduced pressure. The crude solid was dissolved in water (40 ml). After filtration of sn insoluble material, the filtrate was chromatographed on a reverse phase column (packing of PrepPAK/C^g, 100 ml) using water and 5%, 10%, 20% and 30% aqueous CH^OH as eluant. The fractions containing the desired compound were combined, evaporated and lyophilized to give 21 mg (3.8%) of the title compound (I—IH) (E) as a pale yellow powder. Mp. >180°C (dec.). λ -Ji IR : uKBr cm’1 3340(br), 1760, 1670, 1600. max UV : ^Phosphate buffer (pH 7) nm (£l% } 222 (362), 285 ( 452 ) max 1 cm .D_CXNaHCO. HMR :62 Ppm 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH^, 5.33 (IH, d, J=4.5, 6-H), 5.46 (2H, d, J = 7, Ch’ = CH-CH2), 5.90 (IH, d, J = 4.5, 7-H), 6.17 (IH, d-t, J=16 & 7, 3-CH=CH), 7.02 (IH, d, J = 16, 3-CH trans), 8.43 & 9.09 (each 2H, c, J=7, Py-H). h2n OCH. t-1 7-(2-(5-Ami no-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3[3-( 3-am in ome thy lpyr id in io) - I-propen-1-yl ] - 3~ceph em-4-carboxyl at e (:-ii) A mixture of diphenvlmethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido J-3-(3-iodo-l-propen-1y1)-3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmole) and 3-(tbutyloxycarbonylaminomethyl)pvridine (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 HCOOH (5 ml) was stirred at 40-50°C for 50 minutes and evaporated to dryness under reduced pressure. The residual solid was dissolved in water (40 ml), and the mixture was neutralized with NaHCO^. Insoluble material was filtered off, and the filtrate was chromatographed on a reverse phase column (packing of PrepPAK-500/C1θ cartridge, 100 ml), eluting with water, 5%, 10%, 20% and 30% aaueous CH,OH, successively.

J * The fractions containing the desired compound were combined, evaporated and lyophilized to provide 10 mg (1.8%) of the title compound (1-11) (E) as a tan powder.

IR vKBr cm'1 3380(br), 1760, 165Q(sh), 1620(sh) io UV : xfhosphace buffet (pH 7) m (.1S , max 1 cm 235 (sh, 260), 286 ( 370 ) .

NMR : &D2°^NaHC°3 3.68 ( 2H, br, 2-H), 4.16 (3H, s, OCHg), 6.9-8 PPm (IH, d, J=16, 3-CH trans), 8.05 (IH, m, ) Pv-HK), 8.50 (IH, m, Py-H„), 8.80 (2H, m, Py-H 2,6 Example 12 N CONI OCH.

CB«CBCH -CONH. [ 2- ( 5-Amino-l, 2, 4-thiadi.azol-3-yl)-2~methoxyiminoacetamido]-3· ( 3- ( 4-ca rbamoylpyridinio) -1-aropen-l-yl ]-3-cephem-4-carboxyIate ( I-1H) A mixture of diphenylmethyl 7-[2-(5-amino-l,2,4thi adiazol-3-yl)-2-methoxyiminoacetarnido)-3-(3-iodo-1-propen-1yl)-3-cephem-4-carboxylate (IX-1) (E isomer, 4.1 g, 5.7 mmoles) and isonicotinamide (1.4 g, 11 mmoles) in dry DMSO (6 ml) was te ο stirred for 2 hours at room temperature while monitoring by TLC (silica gel plate, CHCl^zCH^OH * 3:1). 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°C for 30 minutes. The resulting solution was concentrated to dryness. The residue was dissolved m H?O (100 ml) and insolubles were removed by filtration. The combined filtrate ano water wash was applied to the top of a column containing reverse phase packing (PrepPAK-500/C^g, 120 ml). The column was eluted with H?O. 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 containing 20% CH^OH). Fraction Nos. 4 and 5 were combined and concentrated to a small volume. Lyophilization gave 25.0 mg (8.1%) of the title compound I~1H, melting at >180°Ο (dec.).

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

Preparation of the hydrochloride - To a suspension of Compound I-1H (98 mg, 0.18 mmole) in CH^OH (1 ml) was added 10% HCl (0.1 ml), and the mixture was stirred for 5 minutes. To the resulting yellow 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 I-1H as a colorless powder. Yield 88 mg (79%). Mp. >190°C (dec.).

IR :υΚΒΓ cm’1 3300, 1770, 1680, 1620 . max UV ; ^Phosphate buffer (pH 7) , • nm ( E 1% ) 227 (385), 266 ma x 1 cm (374) . NMR : 6D2° 2.32 (IH. s, acetone-H) , 3.79 (2H, br-s, 2-H), 4.17 ppm 3H, S, OCHg), 5.34 (IH, d , J- 4.5, 6-H), 5.49 (2H, d 3 = 7, CK=CH-CH2), 5.93 ( 1H, d, 3-4.5, 7-H), 6.28 (IH d-t, 3=16 & 7, 3-CH-CH) , 7.15 (IH, d, 3=16, 3-CH), 8.43 & 9.1 (each 2 K, d. 3 = 7 , Py-H ) . t t Example 13 r-r 7-i 2-(5-Ami no-1,2,4-thia <3 iazol-3-yl)-2-methoxyiminoacetamido]-3[3-(2-methylthiazolio)-1-propen-l-yl]~3~cephem-4-carboxylate (1-13) To a mixture of diphenylmethyl 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido]- 3-(3-iodo-1-propen-lyl)-3-cephem-4-carboxylate (IX-1) (E, 714 mg, 1 mmole) and 2methylthiazole [prepared according to the procedure of R. P. Kurkgy, Ε. v. Brown, J. Am. Chem. Soc., 7 4, 5778 (1952)] (198 nig, mmoles) in dry CH^Cl-, (10 ml) was added AgBF„ (90% ours, 217 mg, 1 mmole) at ~20°C room temperature and with 10% CH,OH-CHC1 !3~" — -3 washed with brine (2 The mixture was stirred for 30 minutes at iltered. The precipitate was extracted (3 x 20 ml). The combined extracts were x 5 ml), dried over MgSO„ and evaporated to ®a * dryness to give a yellow residue, which was triturated with isopropyl ether and filtered to yield 350 mg of the quaternized β 3 product. A mixture of this solid, sodium bisulfite (35 mg) and formic acid (3.5 ml) was mixture was concentrated 0°C for 3 0 minutes. The he formic acid, and the Some insolubles were The filtrate was placed on a reverse stirred at to remove residue was diluted with H^O (40 ml). removed by filtration, phase column (PrepPAK-500/C^g, 100 ml). The column was eluted with H^O (200 ml), 5% aqueous CH^OH (400 ml) and 10% aqueous CHgOH (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^OH). The combined solution was concentrated to a small volume and lyophilized to give 40 mg (7.7%) of the title compound (I-1J) (E). Mp. >195°C (dec.).

IR vKBc cm 1 3300, 1760, 1660, 1600. ma κ UV ^Phosphate buffer (pH 7) max nm (E1% ) 238 ( 442) , 292 (421 ) cm NMR : 0D2O+DMS°-d6 3.06 (3H, s, thiazole-CHg), 3.74 (2H, br-s, ppm 2-H), 4.19 (3H, s, OCHg), 5.92 (IH, d, J=4.5, 7-H), 6.1 (IH, m, 3-CK=CH), 6.8 (IH, d, J=16, 3-CH trans), 8.04 & 8.23 (each IH, d, J = 4, thiazole-H). 9 Example 14 -Ί 7-^2-(5-Am ino-I,2,4-thiadiazol-3-y1)-2-methoxyiminoacetamido]-3[ 3-(4-hyd roxymethvlpyridinio,-1-propen-1-yl]-3-cephem-4-carboxylate (I-IL) k mixture of diphenyl 7-[2-(5~amino-l,2,4-thiaciazol3-y1)-2-methoxyiminoacetam.ido ]- 3-(3-iοάο-1-pr open-l-yl)-3-cephem5 mmole), 4-hydroxy7.5 mmole) in CH^CN (4.5 ml) and CH^OH (3 M_ atmosphere for one 4-carboxvlate ( IX—1) (E isomer,1.07 g, methylpyridine (818 mg, ml) was stirred at room temperature unde hour. After removal o 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, 10 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 atmosphere for one hour. After the addition of 85 % HCOOH (5 ml), the mixture «as stirred under the same conditions for an additional hour. Toluene was added and the reaction mixture was evaporated azeotropicallv under reduced pressure. The residue g of the crude formate this compound (1,15 g) insolubles, which were filtrate and the washes was triturated with acetone to yield 1.17 of the title compound. A suspension of in water (100 ml) was filtered to remove washed with water (10 ml x 2). The were combined and subjected to reverse phase column chromatography The column, which was packed with the packing taken out of a prepPAK-500/C^g cartridge column Ο (Waters) δΟ 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 hy the addition of acetone to give 100 mg of the title compound (X-lL)as a pale yellow powder. To a suspension of the powder (90 mg) in methanol (9 ml) was added 1 H HCl in CH^OH (0.5 ml) and the mixture was stirred at room temperature and concentrated in vacuo. To the concentrate «as added isopropanol to precipitate 77 mg of the hydrochloride of the title compound. .1.0 Pale yellow powder. M. p. >190°C(dec.).

KBr IR : v w cm’1, 1775, 1670, 1635, 1530, max Phosphate buffer (pH?) UV · λ max nm (e), 230 (22600), 264. (sh, 163Ό0) 15 NMR : 6D2° ΡΡΠ' 3.83 (2H, br. 2-CH), 4.17 (3H, s, OCH3), J«4 .5 5.06 (2H, S, N^2y-CH,OH), 5.36 (IH, d, HZ, 6-H), 5.41 (2H, d, J«7 Hz, CH«CH-CH,), .94 (IH, d, J-4.5 Hz, 7-H), 6,36 (IH, d-t, J=16 and 7 Hz, CH»CHCH2), 7,13 (IH, d, J=16 Hz, CH^CH-CH,), 8.08 and 8.83 (each 2H, d, J-7 Hz, Py-a). 1 Example 15 >CONH' OC. sf X^^CBsCHCHu ja 2 CCKj ONH.

I - 2 H * £ -[2-(5-Ami no-1,2,4-thiadiazol-3-yl)-2-ethoxyiminoacetamido]-3i 3-( 4-carbamoylpyridinio)-1-propen-1-yl]-3-cephem-4-carboxylate (I-2H) To a solution of 200 mg of 7-amino~3-[3-(4-carbamovlpyridinio)- 1-propen-l-yl]-3-cephem-4-carboxylate hydrochloride (£ isomer) in 5 ml of 50 % aqueous acetone was added portionwise 190 mg of 2-ethoxyim.ino-2-( 5-am.ino-l, 2,4-thiadiazol~3-yl )acetvl chloride hydrochloride [prepared according to the procedure described in published Japan patent application (Kokai) 57-24389 (2/8/82)], and the mixture was adjusted to pH 6.5-7.0 with 2 N Na^COg (about 1 ml). The reaction mixture was stirred at 10 °C for an hour, acidified to pH 2 with 1 N HCl 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 (20 ml) to give 263 mg (93 I) of the title compound (I~2H). M. p. 170 °C (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 HCl in CH^OH and the mixture was stirred at room temperature for 30 minutes. The solution was filtered and concentrated under reduced pressure. To the residue was added 15 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 %). Μ. p. 160 °C (dec.). Estimated purity 65 %.

KBr IR : v „ cm1, 3300, 1780, 1680, 1620. Πια X UV ; phosphate buffer (pH7) njn (e), ?27 max (22300) , 288 (22800). NMR : 6D2° Ppm 1.44 (3H, t, J«7 3,74 (2H, br. s, Hz, OCH?~CH3), 2-H) 4.45 (2H, q, J=7 Hz, OCH2-CH3), 5.36 (IH, d, J»4.5 Hz, 6-H), 5.46 (2H, d, J«7 Hz, 3-CH=CH-CH2),5.92 (IH, d, J=4.5 Hz, 7-H), 6.20 (IH, m, 3-CH=CH), 7.04 (IH, d, J=16 Hz, 3-CH = CH), 8.43 (2H, d, J=7 Hz, Py-HA), 9.10 (2H, d, J = 7 Hz, Py~Hg).

Example 16 7-(2-(5-Amino-l,2,4-thiadiazoi-3-yl)-2-methoxyiminoacetamido]-3(3-( 4-carbamoylpyridlnio)-l-propen-l-yl )-3-cephem.~4-carboxylate (I-IH) (E isomer ) This Example shows the preparation of Compound I-1H via the last few steps of Reaction Scheme la or lb, wherein the intermediate benzhydryl 7-I2-(5-aminO"l,2,4-thiadiazol-3~yl)-2methoxyiminoacetamido )-3-1 3-(4-carbamoyIpyridinio)-1-propen-1y1]~3-cephem-4-carboxylate formate (XXVU-lH) is isolated. 3 A . Benzhydryl 7-(2-(5-Ami no-1,2 ,4-thiadia2ol~3-yl) -2-methoxyiir:noacetamido]-3-[ 3-( 4-carbamoyl-l-pyridinio)-l-propen-l-yl ] - 3cephem-4-carboxylate Formate (E isomer) (XXVII-1H) A solution of XII-1H (Υθ « ]θ , e isomer) (34 g, 75% pure) in a mixture of acetone and CH,OH (1/1, 20ύ ml) was olacec 3 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 >150eC (dec.). 2R : vKSr cm1 3300, 1780, 1680, 1630, 1600. ma x : λ nm. (c, ) z82 (18o). ma x 1cm -ac etone-d ^/CH-,ΟΗ-d ,(1/1) 0 o 3 4 4.0 (3H, s, OCHg), 5.26, (IH, d «s> ppm. 3=4.5 Hz, 6-H), 5.43 (2H, d, J- 7 Hz, CH-,Ν ' ) 5.99 (IH, d, 3=4.5 Hz, 7-H), 6. 5 (IH, m, 3-CH=CH), 6.92 (IH, s, CHPh2), 7.1 (IH, d, = 16 Hz, 3-CH), 7.35 (10H, m, Ph~H ), 8.36 (IH, s, HCOO), 8.46 & 9.12 (2H each, d, = 8 Hz, Py-H).

B . 7-(2-(5-Am ino-1,2,4-th iadiazol-3-y l)-2-me thoxyiminoacetamido)-3-[3-(4-carbamoyl-l-pvridlnio)-l-propen-l-yl)-3-cephem-4carboxylate (Ι-lH) (E isomer) A mixture of XXVii-lH (E isomer) from Step A (29 g, 75% 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 with acetone (500 ml). The powder was collected by filtration, washed with acetone (2 x 100 ml) and «*« /0 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 HCl (1 l and 0.5 L). The aqueous extracts were combined and placed on a column packed with Diaion HP-20 (1.5 L). The column was washed with water (8 L) and eluted with 30% CK^OH (5 L). 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 CH^OE (100 ml) was added N HCl in CH^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 50 ml of the volume and precipitated with isopropanol (200 ml). The resulting powder was collected, washed with isopropanol (50 ml) and dried in vacuo to give 10.5 g (85% pure) of the title compound I-lH (E isomer) (HCl salt), melting at >180°C (dec.). Pale yellow powder.

Example 17 7- [ 2-( 5-Ami no-1, 2,4-thiadiazol-3-yl)--2-me thoxy iminoacetamido )-3(3-( 4-ca rbamoylpyri din io) -1-pr open-1-yl)-3-ceph em-4-ca rbox ylate ( ϊ-IH) (E isomer ) This example shows the preparation of Compound I-1H via the last few steps of Reaction Scheme la or lb, wherein intermediate XXVH-IH (the formate) is not isolated.

A solution of ΪΧ-1 (E isomer) (27.6 g, 38.5 mmole) and isonicotinamide (22,8 g, 187 mmole) in a mixture of CH^CN (120 ml) and CH^OH (100 ml) was stirred at room temperature for 1 hour under a nitrogen atmosphere. After evaporation of the organic solvents,, the oily residue was triturated with isopropyl ether to give 50.5 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 temperature for 40 minutes and subsequently at 40®C for 1 hour under The mixture was evaporated in vacuo. The residual oil was mixed with toluene (50 ml), evaporated azeottopically and triturated with acetone (400 ml) to give 27.8 g of the crude title compound.

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

The acid extracts were combined and placed on a column of HP-20 resin (1.5 L). The column was eluted with water (9L) and 30% actions containing the desired compound o give a yellow oil, which was methanol (10L). The were combined and concentrated triturated with acetone (300 ml) to yield 9.35 g o) zwitterion form, of the title compound. rhe To a suspension of the product (9.3 g) in CH^OK (ISO ml) was added 1 N HCl in CH,OE (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-1H (Ξ isomer) as its hydrochloride. Paleyellow amorphous powder. M.p. >195°C (dec.).

Example 18 7-(2-( 5-Amin o-l, 2,4-thiadiazol-3-yl )-2~ιη6ϋ1;οχνιΠίίηο3θΘίβΓΡ-ίάο]-3( 3- ( 4-c a r bamoy Ipyr idin io ) -1-propen-l-yl ] - 3-ceph em.-4-ca rboxyl ate ( I - IH) ( E isomer ) This example shows the preparation of Compound I-1K via the last step (7-N-acylation) of Reaction Scheme Ic.

To an ice-chilled suspension or the /-ammo-cephem. hydrochloride XXII-H (E isomer) (5.0 g, 12.6 mmole) in 50% aqueous acetone (100 ml) was added sodium bicarbonate in small portions. The pH of the mixture throughout the reaction. To the about 7) was added 2~(5-amino-1, methoxyiminoacetyl chloride hydr small portions over a period of reaction mixture was maintained occasional additions of sodium b also monitored by tic. After al was monitored by a pH meter cold neutralized solution (pH 2,4-thiadiaxol-3-yl)-2- ©chloride (4.02 g, 15.6 mmole) in an hour, and the pH of the in the range of 6.8-7.5 by icarbonate. The reacti on was of Compound XXII-H had been 6 consumed, the mixture was acidified to pH 3 by the addition of 2 M 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 (1.5 L), 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 (10 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, 5.16 g of the title compound I-1H (E isomer) hydrochloride was obtained as a yellow amorphous powder. M.p. >190cC (dec.). Estimated purity 75% .

Example 19 Purification and crystallization of Compound I-1H (E isomer) Compound I-1H hydrochloride obtained in Example IS 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-,0 and filtered through a celite pad. The ambercolored filtrate (pH 2) was passed through a reverse phase column (the packing of prepPAK-500/C^g cartridge, Waters; 120 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 95%).

To a suspension of the 951 purity powder (3.2 g) in CH ..OH (32 ml) was added N HCl in CH^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 HCl salt (amorphous powder; estimated purity 95%).

A solution of the 95% purity hydrochloride (1 g) in water (4 ml) was adjusted to pH 6.5 with NaHCO^ (200 mg) and stirred for 30 minutes. The crystals which separated during stirring were collected by filtration, washed with water (2 x 5 ml) and dried in vacuo to give 710 mg of I-1H (zwitterion form) as pale yellow prisms. M.p. >185°C (dec.). Microanalysis showed it to be the trihydrate.

IR : vKBr cm"1 1780, 1695, 1660, 1630, 1610. max ,,,. , Phos ©hate buffer UV : λ ( ©H7 ) nm (ε) 227 (22000), 290 (23000). max , DMSO-d_+D.,0 h MR : ο o 2 ppm 3.45 (2H, br, ?-H) , 3 4.99 (IH, d, J-4.5 Hz, 6-H), 5.16 (2H, d, J«=7 HZ, CH2N+), 5.61 (IH, d, J = 4.5 Hz, 75.8 (IH, d-t, J*16 & 7 Hz, 3-CH«CH), 6.93 (IH, d, J*16 Hz, 3-03), 8.18 & 8.89 (each 2H, d, J=7 Hz, Py-H).

Anal. Calc’d for C,ΊH,QNgO6S2'3H^O * Column, Found; C, 42.14; H, 4.38; N, 18.72; s, 10.71. /*· /1 "7 a I * Ur ? ά> β X e* Β, 4.35; N, 18.86; s, 11.00. 10 mm; Mob ile phase, 0.01 M S 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 3 few crystals of the pure 1-1H.

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^ (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 (2x2 ml) and dried under reduced pressure to give 170 mg (80% recovery) of pale yellow prisms of I-1H (zwitterion form), melting at >185eC (dec.), which was identical with that obtained by Procedure 1, (as shown by IR, UV, NMR ;.

The crystalline zwitterion form, of Compound I-1K was slightly soluble in water (6 mg/ml in saline at 23°C). 3 Example 20 I-1K *E 7-(2-(5-Ami no-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetarnido)-3( 3-(3-hyd r oxymethylpyrid in io)-1-propeny1]-3-cephem-4-carboxylate £1- IK ) (E i somer) A . Diphenylmethyl 7-(2-(5-Amino-l,2,4-thiadiazol~3-yl)-2methoxyiminoacetamido]-3-(3-(3-hydroxymethylpyridinio)-lpropenyl)-3-cephem-4-carboxylate iodide (E-isomer) (XII-1K) To a solution of IX-1 (E-isomer, 1.79 g, 2.5 mmoles) in 10 2.5 ml of CHgOH and 7.5 ml of CH^CN was added 3-hydroxvmethylpyndine (545 mg, 5 mmoles), and the mixture was stirred 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-1X as a tan powder. Mp. 170-130°C (dec.).

IR : ν^χ (KBr) in cm"1 1780, 1725, 1675, 1615, 1530, 1385, 1225, 1040, 750, 700.

TV : (C,H5OH) in nm (E1% ) 290 (196). cm NMR i> (DMSO + D2O) in ppm 3.7 (2H, br.s, 2-H), 3.91 (3H, s, OCH3), 4.70 (2H, s, Py-CH^OH) , .28 (2H, m, CH2~N+), 5.23 (IH, d, J«5Hz, 6-H), 5.90 (IH, d, J=5Kz, 7-H), 6.34 (IH, m, 3-CH-CH) , 6.86 (IH, d, U=16Kz, 3-CH), 6.89 (IH, s, CHPh?), 7.35 (10H, m, Ph-H), 7.9-8.9 (4H, m, Py-H).

B . 7-(2-( 5-Ami no-l ,2,4-thiadiazol-3-vl )-2-methoxyiminoacetamido]-3-[3-(3-hydroxymethylpyridinio )-1-propenyl)-3cephem-4-carboxylate (I-1K) (E isomer) A mixture of XII.-IK (E isomer, 2.0 g, 2.4 mmoles) and sodium bisulfite (1 g) in 85% HCOOE (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/C1q cartridge (Waters)) to give 283 mg (22%) of I-1K as an amorphous powder. The powder was crystallized from 4N K2SO^ and acetone to give 144 mg of the title compound I-IK as colorless needles. Mp. 185-188°C (dec.).

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

% UV : λ (Phosphate buffer, pH 7) in nm (E ) 236.5 (283), 1 cm) 275 sh (280) , 292.5 (330).

I N MR : δ (D^O) in ppm 3.75 (2H, s, 2-H), 4.18 (3H, s, OCHg), 4.97 (2H, s, Py-CH2OH), 5.35 (IH, d, J-4HZ, 6-H), 5.43 (2H, d, 3-=6. 5Hz, CH7-N+), 5.92 (IH, d, 3«4Hz, 7-H), 6.10 {IH, d-t, 3=16Hz, 3=6.5Hz, 3-CH=CH-), 6.97 (IH, d, 3=16Hz, 3-CH), 8.13 (IH, d-d, J=8Hz, J=6Hz, Py-H), 8.60 (IH, d, J=8Hz, Py-H), 8.84 (IH, d, 3=6Hz, Py-H), 8.90 (IH, s, Py-H).

Example 21 CONI '©t' cocP CH=CH-CH. 7-(2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(2)-methoxyiminoac efc amido]-3-[3-(4-N-methylcar bamoy1pyridinio)-1-propenyl)-3cephem-4-carboxylate (I-1M) (S isomer) A mixture of diphenylmethyl 7-[2-(5-amino-l, 2,4t hi adiazol-3-yl)-2-methoxyiminoacetamido )-3-(3-iodo-l-pc openy1) 3- cephem-4-carboxylate (IX-1) (E isomer, 450 mg, 0.62 mmole) and 4- N-methylcarbamoylpyridine (prepared according fco the procedure of M. Same^ima, Yakugaku Zasshi, 80, 1706 ( 1960) J (215 mg, 1.58 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 85% formic acid (2 ml) was stirred for 4 hours and then heated at 40*C for 30 minutes. The mixture was evaporated under reduced pressure. The residue triturated with acetone and the crude oroduct was collected Itration. The crude product was chromatographe< on a column was by riAtiesc of HP-20 (1.5 x 18 cm) and the column was eluted with water and % aqueous methanol. The methanolic eluate was evaporated unde 2 reduced pressure and the residue was freeze-dried to give 140 .mg of an amorphous powder, which was further purified by HPLC (Column: Lichrosorb RP-18, Solvent: 15% CH^OH) and the eluate of HPLC was freeze-dried to give 60 mg (18%) of the title product I-1M. Mp. 180-183°C (dec.). Estimated purity: 80%.

IR : v (KBr) in cm 1 1760, 1660, 1600. max UV : λ (Phosphate buffer, pH 7) in nm (e) 230 ( 22100) , 266 max ( 22100 ) .

NMR : 6 (D?O) in ppm 3.06 (3H, s, CONHCH3), 3.72 <2H, s, 2-H), 10 4.16 (3H, s, OCH3), 5.35 (IH, d, J=4.5Hz, 6-H), 5.95 (IH, d, J=4.5Hz, 7-H), 7.00 (IH, d, J=16Hz, 3-CH), 8.35 (2H, d, J-6Hz, pyridine-H), 9.05 (2H, d, J=16K2T pyrIdine-H ), I-lN *E/Z * 7/1 - [ 2 - ( 5-Am.ino-l, 2,4-thiadiagol-3-yl)-2-methoxyiminoacetamido]-3[3-(4-ca rboxypyri dinio )-1-propenyl]-3-cephem-4-ca rboxylat e ( I-IN) To a stirred suspension of isonicotinic acid (340 mg, 2.8 mmoles, in dry DMF (3.5 ml, was added N,O-bis(trimethyl~ silyl,acetamide (0.7 ml, 2.8 mmoles) under nitrogen atmosphere.

To the resulting clear solution was added diphenylmethyl 7-(2(5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido 3-3-(325 iodo-l~pcopenyl)-3~cephem-4-carboxylate (IX-l) (E isomer, 720 mg, mmole) in one portion, and the red solution was stirred for 1.5 hours at room temperature. The reaction mixture was added 3 dropwise to a stirred saturated sodium chloride solution (50 ml) containing sodium thiosulfate (150 mg). The yellow precipitate was collected hy 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 (5 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 cf 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^g cartridge (Water's System 500), and the column was eluted by water (200 ml). The eluent was '5 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 (37%) of the title compound I-1N as a yellow powder. E/2 = 7/1; 80% pure. Mp. >189°C (dec.).

IR : v (KBr, in cm 1 1770, 1665, 1600. ma x UV λ (Phosphate buffer max pH 7) in nm (ε) 227 (22500), 290 (22100).

NMR : ό (D?O NaHCOp in ppm 3.7 d , (2H, br.s). 4.15 ( 3H, s) , (ih, a, 4Hz) , 5.39 (2H =6Hz), 6.14 (IH, d-t, .5 and 6Hz) , 7.03 (IH, a, .5Hz), 8.31 ( 2H, d, J = /Hz 8.94 (2H, d, J=7Hz) Example 23 NIl N CONK H,N N X\x: JocP H. x-xo 7-(2-( 5-Amino-1,2,4-thi ad iazol-3--y 1)-2-(2)-meth oxyi mi nose et amido ) - 3-[3-(2,3-cyclopentenopyr idin io )-1-propenylJ - 3c e oh em-4-c a r b ox y'la te (I -10) (E isomer ) A mixture of diphenylmethy1 7-(2-(5-amino-l,2,4thiadiazol-3-yi)-2-methoxyiminoacetamido]-3-(3-iodo-l-propenvl) 3-cephem-4-carboxylate (IX-1) (E isomer, 450 mg, 0.62 mmole) and 2,-cyclopentenopyridine (217 mg, 1.83 mmole) in acetonitrile (2 ml) 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 65% formic acid (2 ml) was stirred under nitrogen for 3 hours at room, temperature and then heated at 40°C for 30 minutes. The mixture was evaporated under reduced pressure and trituration of the residue afforded 391 mg of the crude product, which was purified by chromatography on a column of HP-20 (1.5 x 18 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 HPLC (Column: Lichrosorb, Solvent: 10% CH^OH). The eluate of HPLC was freeze-dried to give 50 mg (15%) of the title product 1-10. Mp. >190°C (dec.). Estimated purity 75%.

IR : v (KBr) in cm-1 1765, 1670, 1600. max UV : λ (Phosphate buffer, pH 7) in nm (e) 235 (20000), 283 ffiSX (25000).

NMR ό (D2O NaHCOg) in ppm 2.2-2.6 (2H, m, -CH2~), 3.1-3.6 (4H, m, -CH2~), 3.72 (2H, s, 2-H), 4.17 (3H, OCHg), 5.33 (IH, d, JM.5HZ, 6-H), 5.90 (IH, d, J=4.5Hz, 7-H), 6.75 (IH, d, J=16Hz, 3-CH), 7.65-8.2 (3H, m, pyridine-H).

Example 24 I-2N - [ 2 - ( 5-Amino-l ,2,4-thiadiazol-3-yl)-2"(Z)-ethoxyiminoacetamidoj-3-(3-(4-ca rboxypy ridinio)-1-pr openyl)-3-cephem-4cafboxylate (I-2N, E isomer) and 7-(2-(5-Am.ino-l,2,4~thiadiazol~3-yl)-2-(Z)-ethoxyimino15 ac et amido )-3-[3-(4-ca rboxypy r i din io )-I-pr openyl]-3-ceph em-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) (1.0 g, 1.37 mmoles), and the mixture was stirred under nitrogen at room temperature for 2 hours. The mixture was poured into 10% (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 (98%, 5 ml) was heated at 40°C for 1 hour and evaporated under reduced pressure. The residue was triturated with acetone and filtered to give 900 mg of the crude product (E-propenyl isomer:Z-propenyl isomer « 2:1). Separation of the isomers was carried out by HPLC (Column: 8 Lichrosorb, Solvent: HPLC were collected, 151 CH-jOH) The faster moving fractions o and evaporated under reduced pressur freeze-dried to give the E-propenyl isomer of I-2N (44 mg, yield 6%). The slower moving fractions gave the Z-propenyl isomer of I-2N (32 mg, yield 4%) in a similar procedure.

I-2N, i some; Mp.: >20 0°C (dec . ) .

IP. : v (KBr) in cm-1 1765, 1660, 1620, 1380. ma x UV : λ (Water) in nm (ε) 228 (22200), 291 (23600). max NMR : δ 1·^ ( 3H , t, J=6Hz, CH-jCH^), 3.72 (2K, s, 2-H), 4.45 (2H, q, CH2CH3), 5.40 (IH, d J=4Hz, 6-H), 5.90 (IH, d, J=4Hz, 7-H), 7.05 (IH, 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 Mp.: >200°C (dec.) .

IK : v (KBr) in cm 1 1760, 1660(sh), 1620, 1370. ma x UV ; a (Phosohate buffer, pH 7) in nm (ε) 225 (22400), 275 fli S IT * (sh, 16000).

N MR : ό (D,O) in ppm 1.45 (3H, t, J=7Hz, CH-,CH3), 3.50 (IH, d, J«17Hs, 2-H), 3.75 (IH, d, J«17Hz, 2-H), 5.38 (IH, d, J=4Kz, 6-H), 5.9S (IH, d, JMHZ, 7-H), 6.62 (IH, d, J»11HZ, 3-CH), 8.35 (2H, d, J-6Hz, Py-H), 8.92 (2H, d, J"6Hi >y-H) Example 25 Ν· η—C il CONH· JN HjN *0 οοσ- CH«CH-CH2~iT >-CONH_ - [2-( Ami no-1,2,4~thiadiazol-3-y1)-2-(propen-3-yloxyimino)acetamido)-3-(3-(4-carbamoylpyr idin io)-1-propenyl]-3-cepheir-4c a r b ο xylate (1-3H) (Ξ i somer) To a solution of 35 mg (0.03 mole) of 7-amino-3-[3-(4carbamoyIpyridinio)-1-(E)-propenyl]-3-cephem-4-carboxylate hydrochloride in 2 ml of 50% aqueous acetone was added 52 mg of 2- [ 5-amino-l,2,4-thiadiazol-3-yl,] -2- (propen-3-yloxyimino )acetyl chloride hydrochloride (from Preparation No. 25) and the mixfcure was adjusted to pH 6.5-7.0 with 2N Na?CO3> The mixture was stirred at room temperature for 1 hour, acidified to pH 2 with 1 HCI and concentrated under reduced pressure. The residue was chromatographed on a column of HP-20 resin which was eluted with 300 ml of water and 30% CH^OH-HjO. Fractions containing the product were combined and evaporated under reduced pressure. Th residue, 73 mg, was purified by a column of reverse phase carrie which was taken out of a PrepPAK-500/C^g cartridge (Waters, 30 ml). The column was eluted with water, 5% CH^OH, 10% CH^OK and 20% CH^OH, successively. Fractions containing the product were combined and lyophilized to afford 26 mg (62%) of the title product I-3H. Mp. 160°C (dec.,.

XR v (KBr) in cm ma x 3400, 1765, 1680, 1605, 1400.

UV λ (Phosphate buffer, oH 7) in nm (ε) 226 (24600), 288 ΓΠώΧ ( 22 800) . 3 NMR : ό (DjO) in ppm 3..75 (2H, s, 2-H) , 5.41 (IH, d, J«5Hz, 6-H), 5.50 (4H, m, CH,N* t CE^CHp, 5.96 (IH, d, J«5Hz, 7-H), 6.20 (IH, tn, 3-CH-CH), 7.09 (IH, d, J-17HZ, 3-CH), 8.50 (2H, d, J-7HZ, Py-H), 9.16 (2H, d, J«7Hz, Py-H).

Example 26 N· CII N COMB $ \^N:h«ch-ch2?n^~^\- conh.

CH.,c~=cb co-cP I-4H 7-.2-( 5-Amino-l, 2,4-thiadiazol~3-yl )-2-proparqyloxyiminoac etami do )-3-[3-(4-carbamoylpyri din io )-1-propenylj-3-cephem-4carboxylate (I-4H) (E isomer) To a solution of 86 mg (0.19 mmole) of 7-amino~3"[3(4-carbamoylpyridinio)-l-(E)-propenyl)-3-cephem-4-carboxylate hydrochloride (XXII-H) in 2 ml of 50% aqueous acetone was added 63 mg of 2-propargyloxyimino-2~(5-amino-l,2,4-thiadiazol-3-y1)acetyl chloride hydrochloride (from Preparation No. 26). The suspension was maintained at pH 6.5-7.0 with 2N Na2CO3 and then stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 2 with IN HCl and concentrated in vacuo. The residue was diluted with 30 ml of water, neutralized with NaHCO^ and filtered. The filtrate was transferred on the top of a column which was packed with reverse phase carrier (30 ml) taken from a PrepPAK-500/C,θ cartridge (Waters). The column was eluted with water, 5% CH^OH, 10% CH^OH and 20% CH^OH, successively. Fractions containing the product were combined and lyophilized to afford 13 mg (12%) of the title product I-4H. Estimated purity 70%. Mp. 160°C.

IR ; v_ (KBr) in cm'1 3400, 2120, 1765, 1680, 1610. π«·& X UV λ,___(Phosphate buffer, pH 7) in nm (ε) 229 ( 24000), Ind X 288 (21200) NMR ό (D2O) in ppm 3.78 (2H, s, 2-H), 5.15 (2H, d, U-lHz, -CH?-C"CH), 5.40 (IH, d, J*5Hz, 6-H), 5.50 (2H, m, CH-N+), 5.98 (IH, d, U=5Hz, 7-H), 6.20 (IH, m, 3-CH»CH), 7.05 (IH, c, J«17Hz, 3-CH), 8.50 (2H, d, U=7Hz, Py-H), 9.16 (2H, d, J«7Hz, Py-H).

MExample 27 CON ch-ch-ch9-n Ι-5Ϊ * p CONH. 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetamido 1-3-(3-(4-ca rbamoylpyr i din io )-1-propenyl]-3-cephem-4carboxylate (I-5H) (E isomer) To a stirred solution of 139 mg (0.31 mmole) of 7-amino 3-(3-(4-carbamoylpyrid inio)-1-propenyl]-3-cephem-4-carboxylate hydrochloride in 3.5 ml of 50% aqueous acetone in an ice-cooling bath was added portionwise 120 mg (0.44 mmole) of 2-(5-amino1,2,4-thiadiazol-3-yl)-2-cyclopentyloxyiminoacetyl chloride hvdrochloride (from Preparation No. 27). The mixture was (0.9 ml) and stirred fo: adjusted to pH 6.5-7.0 with 2N Na9CO. hour at 10°C. The reaction mixture was acidified to pH 2 with IN HCl and evaporated 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^OH-H^O, successively. Fractions ccntaining the product were combined and concentrated in vacuo. The residue was treated with 60 ml of acetone to give 111 mg (63%) of the title compound Σ-5Η. Mp. 160°C (dec.). Estimated purity 70%. Ο IR : v (KBr) cm ~ m«aX 3400, 1770, 1680, 1605, 1530. UV :Xmax (^osphate buff er, pH 7) in nm (ε) 224 (23300), 286 (24600). N MP : ό (DMSO-dg) in ppm 1.70 (8H, br.s, ), 4.66 ( IH, H \ y br.s, χ ), 5.05 (IH, d, J=5Hz, 6-H), 5.30 (2H, m, CH2n), 5.67 (IH, d-d, J = 5Hz £ 7Hz, 7-H), 6.20 (IH, m, 3-CH=CH), 7.08 (IH, d, J=17HZ, 3-CH) 8.34 (2H, d, J=7Hz, Py-H), 9.11 (2H, d, J=7Hz, Py-H), 9.38 (IH, d, J=7Kz, 7-NH).

Example 28 N. ,1 fi - CONH— I I n 1 J — W fs' OCH cocP CH-COOH I-1P *E 7-(2-(5-Amino-l,2, 4-thiadiazol-3-yl)-2-methoxyiminoacetamido]-3[ 3-( 3-carboxymethylpyridinio)-l~propenyl ]-3-eephem.-4-carboxylat.e (1-1P) (E isomer ) A. DiphenyImethyl 7-(2"(5-Amino-lg2f4-thiadia2ol-3-yl)-2methoxyiminoacetamido)-3-(3-(3-carboxymethylpyridinio)-1propenyl)-3-cephem-4-carboxylate (XII-1F, iodide, £ isomer) To a suspension of 3-carboxymethylpyridine hydrochloride (0.89 g, 5 mmoles) in 10 ml of CH?CI, was added Ν,Ο-ois(trimethylsilyl)acetamide (4.97 ml, 18 mmoles), and the mixture was stirred at room temperature until a clear solution was obtained. To the solution was added IX-1 (1.79 g, 2.5 mmoles), and the mixture was allowed to stand at room q ./ /t emperature .

After 3 hours, 3 ml of CH^OH was added to the cooled mixture and the solution was evaporated in vacuo to give an o f oil which was triturated with the title compound XII-1P as a ethyl tan acetate to powder. Mp. afford 161°C 2.28 g (dec .) 5 :r : v (KBr) in cm1 1780, HIS X 1720, 1675, 1630, 1530, 1385, 1225, 1045, 755, 700. uv (C-HcOK) in nm (E1% rn & X 2 5 ) 295 (188) . 1 cm N.’IR : ό (DMSO * I^O) PPm 3.70 (2H, br.s, 2-H), 3.90 (5H, s, OCH3 & Py-CH-CO), 5.25 (3H, m, -CH2N’ & 6-H), 5.92 (IH, d, J=4.5Hz, 7-H), 6.35 (IH, m, 3-CH=CH-), 6.90 (IH, d, J=16Hz, 3-CH), 6.92 (IH, S, CHPh,), 7.35 (10H, m, Ph-H), S.8-9.0 (4H, m, Py-H). 3. 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacet amido]~3-[3-( 3- ca rhox yme thyl pyridin lo)-l~pr openyl]- 3cephem-4-carboxylate {I-1?) (Ξ isomer ) A mixture of XII-1P (iodide) (2.28 g) and sodium bisulfite (1.1 g) in 85% HCOOH (10 ml) was stirred at room temperature for 2 hours. The reaction mixture was concentrated tc 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 I-IP (38%, from IX-1) as a pale yellow amorphous powder. Mp. 165®C (dec.), IB : v (KBr) in cm1 1770, 1670, 1600, 1530, 1385, 1140, ma x 1040 . ¢1 9 UV : X (Phosphate buffer, pH 6.2) in nm (Ξ max cm ) 234 (374), 277sh (390), 290 (402).

NMR (0^0 + MaHCO-j) in ppm 3.78 ( 2H, s, 2-H), 3.92 (2K, s, Py-CH2C0), , 4.22 (3H, s, OCHg), 5.40 ( IH, d, J-4HZ, 6-H), 5.44 ( 2H, d, J«6.5Hz, -CH?-N+), 5.97 ( IH, d, 1-- UHz, 7-H), 6.20 (IH, d-t, J=16 £ 6.5Hz, 3-CH = CH), 7.08 (IH, d, J=16Hz, 3-CH), 8.11 ( IH, d-d, J=8 £ 7Hz, Pv-Hg), 8.53 ( IH, d, J=Sh’z, Py-H4), 8.82 (IH, d, J= 7Hz, Py-Hg), 8.86 (IK, s, Py~H2 ) .

Example 29 S N-Cι y < OCH•CONH/COiP X © ΖΓ^λ CH«CH-CH,-N7 yS-CH-jCOOH Ϊ-IQ *E 7-12-(5-Amino-l,2,4-thiadiagol-3~yl)-2-me thoxyiminoacetarnido]-3[3-(4-carboxymethylthiopyridinio)-l-propenyl)~3-cephem-4ca rboxvlate (I—IQ) (E isomer) A· DiphenyImethyl 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2~ methoxyiminoacetamido]-3-(3-(4-carboxymethylthiopyridinio)I-propenyl1-3-cephem-4-carboxylate (XII-1Q, iodide, E isomer ) To a suspension of 4-carboxymethylthiopvridine (0.88 g, mmoles) in 10 ml of CH^Cl^ was added N,O-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-1 (S isomer 1.79 g, 2.5 mmoles), and the mixture was '3 allowed to stand at room temperature. After 3 hours, 3 ml of CH.OH was added to the cold mixture and the solution was 3 evaporated in vacuo to give oily residue which was triturated with ethyl acetate to give 2.43 g of the title compound XII-1Q (iodide) as a tan powder. Mp. 155°C (dec.).

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

UV ς___(C0HcOH) in nm (E1% ) 312 (299). maX 2 5 . cm NMR ; 6 (DMSO-dg + D2O) in ppm 3.70 (2H, br.s, 2-E), 3.93 (3H, s, OCH3), 5.07 (2E, m, N+), 5.23 (IK, d, J=5Hz, CH 6-H), 5.90 (IH, d, J=5Hz, 7-H), 6.29 (IH, m, 3~CH=CK), 6.87 (IH, d, J=16Hz, 3-CH), 6.91 (IK, s, CHPh2), 7.35 (10H, m, Ph-K), 7.86 & 8.56 (each 2H, d, J=6Hz, Py-H).

B . 7-(2-(5-Amino-l,2,4~thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[3-(4-ca rboxymethylthiopyridinio)-1pcopenyl]-3-cephem-4-carboxylate (I-1Q) (£ 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 hours. The reaction mixture was concentrated tr 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 g), which chromatography (HP-20, 20 ml) compound I-1Q (39% from IX-1) Mp. Ιθδ’Ο (dec.). was purified by column to afford 577 mg of the title as a pale yellow amorphous powder (KBr) in cm"1 1765, 1670, 1625, 1530, 1380, 1110, 1035 . 4 ι % UV : λ (Phosphate bufrer, pH 6.2) in nm (EA ) 234 (459), filcsX . 1 310(678).

NMR : ό (D^O + NaHCOj) in ppm 3.79 (2H, br . s, 2-H), 4.10 (2H, s, S- -CH? ) , 4. 23 (3H, s, OCH ) , 5 5.25 ( 2H, d, J =3 6.5Hz , CH?-N'T) , 5.39 (IH, d, . 0Hz, 6-H), 5.97 (IH, d, J »4Hz , 7-H), 6.18 (IH, d-t, J«15 . 5Hz & 6.5Hz , 3-CH=CH), 7.05 ( IH, d, J=15.5Hz , 3-CH), 1.0 7.84 & 8. 55 ( each 2H, d, J - 7 K Z , Py-H) . 3CX@/~1 CH2-N Ϊ-IA *E/2 - 7/1 7-(2-( 5-Ami no-1,2,4-thiadiazol-3-yl)-2-methoxyiminoacetamido) 1 5 3-(3-(l-methvIpyr rolid ini o)-1-propenyl3-3-cephem-4-carboxylate sulfate (I-1A, sulfate) A. Diphenylmethyl 7-(2-(5-Amino-1,2,4-thiadiazoI~3-yl)~2m.e thoxy imi noace tamido 3-3-[3-(1-methyIpyrrolidinio)-1propenyl3-3"Cephem-4-carboxylate (XII-1A, iodide) To a cold solution of diphenylmethyl 7-[2-(5-aminol,2,4-thiadiazol-3-yl) -2-me thoxy imi noace tamido 3 - 3- (3-iodopropenyl)-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-methylpyrrolidine (2.55 g, 30 mmoles) in ethyl acetate (30 ml) over a period of 1 hour at -5 to 0°C, with stirring. After stirring for an additional 10 minutes, the resulting precipitate was collected by filtration and washed with chloroform (200 ml) to give 23.0 g (95.8%) of the title compound (IX-1A, iodide), melting at >175°C (dec.), IF : υ (KBr) in cm1 3300, 1780, 1730, 1685, 1615. ΙπαΧ UV : λ _ (C^HeOH) in nm (E1% ) 218 (435), 295 (188). max 2 5 i cm B. Diphenylmethyl 7-[2-(5-Amino"I,2,4-thi3dxazoI~3-yl)-2 methoxyiminoacetamido]-3-[3-( l~methylpyrrolidinio)-lpr openyl)--3-cephem-4-ca rboxyl ate (XII-1A, chloride) The compound (XII-1A, iodide) (23 g, 28.7 mmoles) was dissolved in a mixture of acetone and methanol (1:1, 230 ml) and applied on an 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 and concentrated to an oily residue, which was triturated with ethyl acetate (300 ml) to yield 17.9 g (87.7%) of the title compound (XII-1A, chloride), melting at 190°C (dec.).

IR : υ (KBr) in cm’1 3380, 1780, 1680, 1620. ma x UV : Xmax (C,H5OH) in nm (E1% ) 220 (369), 290 (232).

C . 7-1 2-( 5-Amino-I, 2, 4-thiadiazol-3~yI) - 2-me thoxy i rr.inoacetamido]-313-(l-methylpyrrolidlnio)-l-propenyl]-3cephem-4-carboxylate sulfate (I-1A, sulfate) A mixture of the compound (XII-1A, chloride) (17.8 g, mmoles) xn 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-1A. 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 2Ν HCl (1 L and 0.5 L) . The combined extracts were adsorbed on a Diaion HP-20 resin (1.5 L column), which was eluted with water and 30% aqueous methanol. The desired fractions were collected and 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^θ cartridge (90 ml), using water and 10% aqueous methanol as an eluent. The eluate was collected in 20-ml fractions with monitoring by HPLC. [Column, Nucleocil SSC-ODS-262, 8 x 100 mm8· Mobile phase, 0.01M phosphate buffer (pH 7.2)/CH3OH « 90:10; Detection, OV (254 nm)]. 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 1], Fraction No. 11 through Fraction No. 85 were worked up in the same manner as described above to give 3.27 g of yellow powder (E/2, « 5/1, 70% pure) (Crop 23. A portion of Crop 1 (1.0 g) was purified by rechromatography on the packing of PxepPAK-50 0/C,θ 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 with acetone (315 7 rr 1), with stirring. The creamy precipitate was collected hy filtration to obtain 1.48 g of the title compound (I-1A, sulfate) ( E/2 « 7/1, 80% pure), melting at >185°C (dec.).

IR : (K3r ) in cm1 3380, 3000, 1765, 1675, 1630, 1535, 1390, 1115.

UV Ii MR ma x (Phosphate buffer, pH 7) in nm (e) 236 (19900), 291.5 (22500) _^S? o (^2° + NaHCOp in ppm 2.36 (4H, br., ), 3.15 ), 3.62 (SB, ), 3.83 (IH, br., 2-H), 4.13 (2K, d, J=8Hz, CH_N+), 4.22 (3H, s, OCH3), 5.39 (IH, d, J=4.5Hz, 6-H), 5.96 (IH, d , J ss 4.5 η z , 7-H), 6.00 (IH, m, 3-CH-CH), 6.67 (1/6H, d, J=10Hz, 3-CH, cis), 7.04 (7/6H, d, J«16Hz, 3-CH, trans).

Example 31 - [2-(5-Amino-1,2,4-thiadia2ol-3-yl)-2~methoxyiminoacetamido]3-[3-trimethvlammon io-l-oropenyl]-3-cephem-4~earboxylate (1-ID ) A . DiphenyImethyl 7-[2-(5-amino-l,2,4-thiadiazol~3-yl)-2~ methoxyiminoacetamido]-3-(3-trimethylammonio-1-propenyl)3-cephem-4-carboxylate (XII-1D, iodide) To a solution of 13.0 g (19 mmoles) of diphenylmethyl 7-[2~(5-amino-1,2,4-thiadiazol-3-yl)-2-me thoxyiminoacetarnido]-3(3~iodopropenvl)-3-cephem-4-carboxylate (IX-1, 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 -5®C, and the mixt ure was stirred for 1 hour at -5 °C. The resulting prec ipi tate was filtered off, washed well with CHCl^ and dried t give 12 . 5 g (88%) of the title compound (XII-lD) as the iodide. IR V max (KBr) in cm1 3300, 1765, 1720, 1665. UV » Λ A max (C HrOH) in nm (ε) 300 (18400). Λ> Ο B. Diphenylmethyl 7-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido)-3-(3-tr imethylammon io-l-propenyl) 3-cephem-4-carboxylate (XII-lD, chloride) The iodide (XII-lD, 12.5 g) was dissolved in 60 ml of CH^OH-aeetone (1:1) and passed through a column of ion-exchange resin [IRA-410 (Cl ), 125 ml]. The column was eluted with 300 m of CH-jOH-acetone (1:1), and the eluate was evaporated in vacuo and triturated with 300 ml of isopropyl-ether to afford 10.4 g (91%) of the quaternary salt (XII-1D, chloride). 9 IR max (KBr) in cm* 3300, 1765, 1710, 1665 UV (C-,Η.ΟΗ) in nm (ε) 298 (151C0) ma x 2 5 C. 7 -[ 2- ( 5-Am.ino -1, 2, 4-thiadiazol-3-y1)-2-methoxyiminoacetamido]- 3-[3-t r imethylammonio-l-propenyl]-3-cepher 4-c a rboxyla t e (I-ID, sulf ate, B 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 HR-20 (300 ml) which was eluted with water (1000 ml), 10% CHgOH (200 ml) and 30% CHgOH (150 ml), successively. Fractions containing 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/C1θ cartridge (Waters, 200 ml). Elution with water (600 ml) and 30% CHgOH (200 ml), successively, followed by concentration of fractions containing the desired product gave 2.52 g (18%) of the title compound. A solution of the zwitterionic oroduct (1.5 g) in IN 2,50., (5 ml) was added .portionwise to 300 ml of acetone and the resulting precipitate was filtered and dried. Yield of I-ID sulfate «as 1.42 g (80%). The ratio of E/2 was aooroximately 10/1 based on HPLC. v . „ (KBr) in cm iTUi X 3380, 1765, 1665 UV ma: (Phosphate buffer, pH 7) in nm (e) 237 (19500), 293 (22400).

IR i 9 ο NMR : δ (D7O) in ppm 3.25 ( 9H , S, N+-CH3), 3.94 ( 2H, s, 2-H), 4.14 (2H, d, J«7Hz, CH?N+), 4.23 (3H, s, O-CH3), 5.42 (IH, d, J«4.5Hz, 6-H), 6.00 (IH, d, J»4.5Hz, 7-H), 6.23 (IH, d-t, J»7 & 16Hz, 3-CH=CH), 7.23 IIH, d, J-16HZ, 3-CH).

H2N c N 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-me thoxyiminoacetamido]10 3-(3-(4-carbamoylpyridinio)-l-propenyl]-3~cephem-4-carboxylate (I-1H, E isomer ) To a mixture of 7-amino-3-(3-(4~carbamoylpyridinio) 1-(E)-propenyl]-3-cephem-4-carboxylic acid hydrochloride (XXIl-H, 397 mg, 1 mmole) and NaHCO^ (168 mg, 2 mmoles) in aqueous DMF (water, 3.5 ml and DMF, 7.5 ml) was added benzotriazol-l-yl-2( 5-amino-l,2,4-thiadiazol-3-yl)-2-methoxyiminoacetate ( 479 mg, 1.5 mmoles) (from Preparation No. 28). The mixture was stirred at room temperature for 3.5 hours. The reaction mixture was adjusted to pH 3-4 with 3N HCl and diluted with 200 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 pH 6.8 with NaHCO^, treated with decolorizing carbon, concentrated to ca. 1 ml and seeded with a few pieces of crystalline Σ-1Η. After stirring overnight, the crystalline precipitate was collected by filtration to afford the title compound I-1H (zwitterion form). Yield 83 mg (16%). Mp. >185°C (dec.). Physico-chemical data of this product were identical to those of the compound in Example 10.

Preparation No. 1 Diphenylmethyl 7-[2-(5~Amino-I,2,4-thiadia2ol-3-yl)-2irethoxyinunoacetemido)-3~chloromethyl-3-cePhem-4carboxylate (IV-1) J To a stirred suspension of 2-(5-amino-l,2,4-thiadiazol3-yl)-2-methoxyiminoacetic acid (Ill—1) (2.1 g, 10 mmole) in dry CH^Cl^ (50 ml) was added PCI,. (2.09 g, 10 mmole) at -30°C, and the mixture was stirred for 20 minutes at -15 to -20 °C. To the above acid chloride solution was added a solution of diphenyl10 methyl 7"amino-3-chloromethyl-3-cephem~4~carboxylate hydrochloride (II) (4.5 g, 10 mmole) in CH^Cl, (50 ml) containing Ν,O-bis~(trimethylsilyl) acetamide (10 g, 50 mmole) at -30 °C, After stirring at -10 aC for 1 hour, the mixture was concentrated to remove the CH2C1-, and diluted with ethyl acetate ( 200 ml).

The mixture was washed with 10 % aqueous NaHCO^ (2 x 40 ml), H?O (2 x 20 ml) and brine (10 ml), successively, and dried over MgSO^. The solvent was evaporated in vacuo and the resulting oily residue (10 g) was dissolved in CHCl^ (20 ml) and chromatographed 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 % CH3OH~CHC13.

Fractions containing the title compound were evaporated to give .7 g (95 %) of XV-1 as a yellow amorphous powder. Μ. ρ. >140 "C (dec.).

IR : UV vKBrcm_1 3300, 1780, 1720, 1680, 1620. ma x .EtOH NMR : PP^ max .DMSO-dg 245 (1800), 280 (9900). .53 (2S, ABq, 2-H), 3.94 (33, s, OCH3), 4.42 ( 2H, s, 3-CH2), 5.22 (IH, d, J«4.5, 6-H) , 5.92 ( IH, d-d, .5 & 6, 7-H) , 6.93 (IH, s, CHPh,) 7.36 (10H, m, Ph-H), 8.1 (2H, br-s, NH,) , 9.58 ( IH, d, J-6, 7-NH). <> Ρ repa r at i on No . 2 Diphenylmethyl 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido)-3-iodomethyl-3-ceph em- 4-ca rboxyl at e (V -1) A mixture of IV-1 from. Preparation No. 1 {5.7 g, 9.5 mmole) and Nal (4.3 g 29 mmole) in dry acetone (50 ml) was stirred for 5 minutes at room temperature. The mixture was concentrated under reduced pressure and the resulting oil was shaken with a mixture of ethyl acetate (100 ml) and H^O (10 ml). The organic layer was separated and washed with 10 % w/v sodium thiosulfate and brine, successiveiy. After drying, the ethyl acetate was removed in vacuo to give 6.1 g (93 %) of the title compound (V-l) as a yellow amorphous powder melting at > 120 °C (cec . ).

IR : vKBrcm-1 3300, 1780, 1725, 1680, 1620. max UV : XEt0Hnm (e) 245 (17000), 282 (12000). max NMR : 6DMSO-d6 3.72 ( 2H, ABq, 2-H), 3.94 (3H, S, OCHp, 4.23 ?pIP {2H, s, 3-CH2), 5.21 (IH, d, J=4.5, 6-H), 5.89 (IH, d-d, J=4.5 & 6, 7-H), 6.94 (IK, s, CHPh^, 7.35 (10H, m, Ph-H), 8.12 (2H, br-s, NH-,), 9.65 (IE, d, J=6, 7-NH).

Preparation No. 3 Diphenylmethyl 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2methoxyiminoacetamido]-3-trlphenylphosphon iomethyl-3-cephem-4carboxylate iodide (VI-1) A mixture of V-l from. Preparation No. 2 (690 mg, 1 mmole) and triphenylphosphine (786 mg, 3 mmole) in ethyl acetate (20 ml) was stirred overnight at room temperature. The solid which separated was collected, washed with ethyl acetate (2 x 10 j. Ο 3 ml) and dried to give 950 mg (100 %) of the phosphonium iodide VI-1. M. p. 186 °C (dec.).

IR uv NMR vn r Ί V °‘cm ~ 3300, 1780, 1710, 1680, 1610. max Ε* «- O X nm (ε) 268 (15000), 275 (13000), 300 (7300). ma x 0DMSO-d6 3.52 (2H, br„Sf 2H), 3.94 (3H, s, OCHg), 5.34 ppir (IH, d, 3=4.5, 6-H), 5.9 (IH, m, 7-H), 6.3 (IH, s), 7.3 (10H, m, Ph-H), 7.8 (15K, m, Ph-H).

L0 Preparation No. 4 D .·, ph eny lme thyl 7-(2-( 5-Amino-l ,2,4-thiadiazoI-3-yI) -2-methoxy lL'in oacetamido ) -3- ( (tr iphenylphosphoranvlidene ) me thyl ) -3c>-phem-4-carboxylate (VII-1) A mixture of VI-1 from Preparation No. 3 (952 mg, 1 5 mmole), Amberlite IRA-410 (OH form, 500 mg) and N NaOH (4 ml) in CH?Cl7 (10 ml) was stirred for 1 hour at room temperature. The mixture was filtered and the separated organic layer was dried over MgSOz and concentrated under diminished pressure. The eg resulting oil was triturated with ethyl acetate and the resulting !0 yellow precipitate was collected by filtration to give 740 mg (90%) of the title compound VII-1. M.p. >180°C (dec.).

I?. : vKBr cm"1 3400, 1750, 1630 . max UV : XEt0Hnm (e) 268 (12000), 276 (10000), 384 (23000). max 104 Preparation No. 5 Diphenylmethyl 7-[2-(5-Amino-l,2,4-thiadiazol-3-yl)~2-methoxyiminoacetamido]-3-(3-chloro-l-propen-1-yl) -3-cephem-4-carboxyla te (VI ΣΙ-l ) To a solution of Vll-i from Preparation No. 4 (6.9 g, 8.4 mmole) were added MgSO„ (3 g) and 40% chloroacetaldehyde (£10 mg, 8.4 mmole). The mixture was stirred for 1.5 hours at room temperature and then filter.ed. 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^, and CHCl^ containing CH^OH. fractions containing the desired product (0.5 - 1% CH^OH-Ch’Cl^) were evaporated in vacuo to give 1.6 g (30%) of the title compound viii-l as a yellow amorphous powder, which was a mixture of the 2 and E isomers with respect to the chloropropenyl moiety (2/3=2/1, by nmr). M.p. >130°C (dec.).

IR : vK3r cm 1 3300, 1780, 1725, 1680, 1620. max UV : x£t0Hnm (ε) 240 (20000), 286 (12000). max NMR .DMSO-d, + D?O 3.56 6 3.8 (m, 2-H), 3.94 (3H, s, OCH^, ppm 4.16 (d, J=7.5, CH?C1), 5.26 (1H, d, J=4.5, 6-H), 5.87 (IH, d, J»4.5, 7-H), 6.28 (2/3H, d, J-ll, 3-CHcis-H), 6.72 (1/3H, d, J=16, 3-CH trans-H), 6.81 (2/3H, S, CHPh?), 6.92 (1/3H, S, CHPh2), 7.4 (10H, m, Ph-H). '' 1 0 s Preparation No, 6 D ipheny Lmethy 1 7-Benzylideneamino-3-[(triphenylphosphoranylidene )methyl )-3-cephem-4-carboxylate (XVI) To a solution of diphenvlmethyl 7-henzylideneamino-3I (triphenylphosphonioJmethyl]-3-cephem-4-earboxylate iodide (XV) [prepared according to the procedure of Japan published patent application (Kokai) 56-86187 (7/31/81)] (60 gf 70 mmole) in CH2C19 (350 ml) were added N NaOH (140 ml) and Amberlite IRA-410 (OH** form, 35 g) at 5°C, The mixture was stirred for 1 hour at 5eC and filtered. The organic layer was separated,, dried over MgSO„, 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 (94%) of the title compound XVI, melting at 195-6®C (dec.). i IR : vK3r cm-1 1770, 1620. max Preparation Mo. 7 Diphenylmethyl 7-3enzyiideneamino-3- (l-chloro-^l-pr open-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,C1? (40 ml) and (10 ml), was added anhydrous chloroacetaldehyde (800 mg) at room temperature. To the mixture was added additional 800 mg of chloro15 minutes, the aqueous layer was removed and the organic was dried over MgSO„. Evaporation of the solvent gave a After drying over MgSO4, removal acetaldehyde 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. Af te: layered oil which was dissolved in a mixture of ethyl acetate and isopropyl ether (1/2, 80 ml). The solution was washed with saturated aqueous NaHCO^ (10 ml) and K9O (10 ml), successively. ir ded 3.3 g of the solvent a; yellow oil. A solution of the oil in CH?C1, (50 ml) was filter· 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^Cl, (50 ml). The filtrate and washing were combined and evaporated to dryness. The residue was triturated with n-hexane to give 1.7 g (80%) of the title compound (XVU) as a yellow powder. The nmr spectrum indicated that the chloropropenyl moiety had the Z configuration. M.p. >50°C (dec.).

IR : vKBr cm-1 3400, 1775, 1720, 1630. max UV : XEfc0Hnm (e) 253 (11000), 258 (11000), 265 (10000), 273 max (8300), 281 (7000), 290 (6300).

NMR : 0DMS0_d6 3.63 (2H, br-s, 2-H), 4.0 (2H, m, CH,-C1), 5.42 ppm (2H, m, 6-H £ 3-CH=CH), 5.72 (IH, d, J~4,5e 7-H), 6.27 (IH, d, J=ll, 3-CH), 6.85 (IH, s, CH?h7), 7.33 (10H, 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 layer (the upper layer) was separated and diluted with CHClj (100 ml), mixed with KgSO^ (20 g), heated to reflux for 5 minutes, and filtered. The solvent and water were removed azeo( ) tropically (b.p. 56-64*0 , and the residue was distilled to give anhydrous chlor©acetaldehyde , b.p. 70-82®C/760 mm.

IR : vfllin cm"1 1720. max (1) R.P. Kurkjy, Ε, V. Brown, J. Amer. Chem. Soc., 7 4, 5778 (1952). (2) S. Trippett, D. M. Walker, J. Chem. Soc., 1961 1266. (3) Η. 0. House, V. κ. Jones, G. A. Frank, J. Org. Chem., 29, 3327 (1964).

Preparation No. 8 Diphenylmethyl 7-Amino-3-(3-chloro-1-propen-1-yl) -3-cephem-45 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 [(cacboxyrcethyl)trimethylammonium chloride hydrazide] (251 mg, 1.5 mmole) in CH^OH (10 ml) containing acetic acid (0.25 ml), at 5*C. After stirring for 30 minutes at 5°C, the mixture was concentrated to remove the CHgOH and then ethyl acetate (20 ml) was added. Th® ethyl acetate solution was washed with H^OJ 2 x 5 ml), saturated aqueous NaHCOg (5 ml) and brine (5 ml), successively and dried over MgSO,. Evaporation of the solvent gave 145 mg (97%) of the title compound XVIII (2 isomer) as a yellow powder. M.p. >100°C (dec.).

IR : vKBr cm1 3400, 1770, 1720. max UV : xStOH nm (e) 252 (3700)? 258 (3800), 260 (4000), 274 (4000), 285 (4000).

Pr eparation No . 9 Diphenylmethy! 7-(2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-methoxyi mi noace tamido 3- 3-( 3-chlor o-1-propen-1-v 1) - 3-cephem-4ca rboxylate (Vii1-1) A mixture of 2-(5-amino-l,2,4-thiadiazol-3-yl)-2m.ethoxyiminoacetic acid (III-l) (10.1 g, 50 mmole) and PC(10.4 g, 50 mmole) in dry CH2C1-, (100 ml) was stirred at -7 to -15°C for 2 hours. The clear solution was poured into n-hexane (500 ml) to give a precipitate. The organic layer was discarded by 3 decantation and the remaining solid was triturated with n-hexane (100 ml). The yellow precipitate was collected by filtration and dried in vacuo to give 12.5 g (99%) of the acid chloride, melting at 8Q°C (dec.).

IR : vnu301 cH 1770. max The acid chloride (25 mg, 0.1 mmole) was added to a solution of XVlil (2 isomer) from Preparation No. 8 (44 mg, 0.1 mmole) in dry CH,Cl, (5 ml) at room temperature, with stirring. After 30 minutes, the mixture was concentrated under reduced pressure and diluted with ethyl acetate (20 ml) and saturated aqueous NaHCO^ (5 ml). The organic layer was washed with saturated aqueous NaHCO^ (5 ml), brine (5 ml), 10% HCl (5 ml) and brine (5 ml). The solvent was dried over MgSO. and then evaporated to dryness to give the product as a yellow foam. The foam was purified by silica gel (Wakogel ¢-200, 1 g, containing 0.1 ml of 1/1.5 M phosphate buffer pH 6.4) column chromatography by elution with CH,C1,~CK3OH (100 : 1), to give 31 mg (50%) of the title compound VilJ-1 (2 isomer) as a yellow powder. M.p. >150°c (dec.).

IR : vKBr cm1 3400, 1775, 1720, 1675, 1630. max U V : nm max (e) 240 (17000), 280 (10000). NMR : .DMSO-d, δ o 3.6 (2HS, m, 2-H) , 3.92 (3H„ s, O-CH3), 4.0 (2H, ppm m, CH,C1), 5.27 (2H, m, 6-H & 3-CH-CH/, 5.83 (IH, d-d, J®4.5 & 10, 7-H), 6.25 (IH, d, J-ll, 3-CH), 6.83 (1H,S, CHPh,), 7.33 (10H, m, Ph-H), 8.0 (2H, br-s, NH,), 9.57 (IH, d, J=10, 7-NH).

Preparation No. 10 Di ph eny lmethy 1 7-(2-( 5-Amino-l ,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido]-3-(3-iodo-l-pgopen-l-yl)-3~eephem-4-earhoxylate (IX-1) A solution of VIII-1 from Preparation No. 5 (2/E-2/I, 430 mg, 0.77 mmole) 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 acetate (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*. Evaporation of th< solvent gave 540 mg (98%) of the title compound IX-1 (2/E=l/l) as a reddish amorphous solid, melting at >120°C (dec.).

IR : KBr -1 υ cm 3300, 1780, 1720, 1630, 1620. max UV : λ EtOH λ nm (e) 240 (21000), 290 (12000). max NMR : ,DMS0vD-0 o 2 3.67 (2H, m, 2-H), 5.29 (IH, d, d 36 4 .5 i , 6-H), ppm 5.95 (IH, d, J«4.5, 7-H), 6. 27 ' (1/2H, d, , 3-CH cis), 6.72 (1/2H, d e 0 = 16, 3-CH CHPh2,, trans), 6.87 & 6.96 (each 1/2H, s, 7.4 (10H, m, Ph-H).

Preparation No. 11 Diphenylmethyl 7-(2-(5-Am ino--1,2,4-thiadia2ol-3-yl)-2-methoxyiminoacetamido)-3-(3-iodo-I-propen-l-yI)-3-cephem-4-carboxylate (IX-l) A mixture of VIII-1 (2 isomer) from Preparation No. 9 (5.6 g, 9 mmole) and Nal (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 (90 ml). The ethyl acetate layer was washed with 10% w/v aqueous sodium thiosulfate solution (10 ml) and B^O (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 (67%) of the title compound IX-l as the Ξ isomer. M.p. >165°C (dec.).

IR : vK3r cm1 3400, 1780, 1725, 1680, 1610. max UV ; XEt0H nm (ε) 240 (18000), 297 (11000). max NMR : 6DMSO~d6+D2° 3.90 (3H, s, OCH3), 5.25 (IH, m, 6-H), 5.95 ppm (IH, m, 7-H), 6.72 (d, J«16, 3-CH trans), 6.96 (IH, s, CH-?h2), 7.4 (10H, m, Ph-H).

Preparation No. 12 Benzhydryl 7-Amino-3-(3-chloro-1-propen-l-yl)-3-cephem-4carboxylate (2-isomer) (XVIII) Compound XVIH is the common intermediate utilised in Reaction Schemas lb and Ic.

A. Benzhydryl 7-Benzylideneamino-3-triphenylphosphoniomethyl-3cephem-4-earboxylate chloride (XV) To a suspension of benzhydryl 7-am ino-3-chloromethy1-3cephem-4-carboxylate hydrochloride (II hydrochloride) (200 g, 0.44 mole) in CH^Cl^ (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^ (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?C1? (200 ml). To tne combined filtrate and washings was added triphenylphosphine (126 g, 0.45 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 (96%).

M.p. 185-190°C (dec.).

IR : vK3r cm1 1780, 1720, 1630. max UV : XCH2C12 nm (ε) 260 (24100). max S. Benzhydryl 7-Benzvlideneamino-3-((triphenylphosphoran· ylidene)methyl3-3-cephem-4-carboxy!ate (XVI) A mixture of XV (322 g, 0.42 mole) and 5 N Na^CO^ (252 mi) in CHjCl, (1.6 L) was stirred vigorously for 15 minutes at room, temperature. The organic layer was separated, dried over MgSO. and concentrated to about 500 ml of volume. The concentrate was added to acetone (1 L), with stirring, to give a light yellow crystalline powder which was collected by filtration to yield 237 g (78%) of XVI, melting at 195-198°C (dec.).

IR : KBr v cm_1 1770, 1620. max UV : XCH2C12 nm (ε) 254 (23000), 389 (22000). max NMR :6CDC13 2.56 & 3.16 (2H, ABq), 5.00 (IH, d, 3 = 4 Hz), 5.23 ΡΡΠ’. (IH, d, J=4 Hz), 5.47 (IH, d, J*22 Hz), s), 7.2-7.8 (30H, m), 8.55 (IH, s). 6.95 (IH, C. Benzhydryl 7-Amino-3-[chloro-1-propen-l-yl)-3-cephem-4carboxylate hydrochloride (2 isomer) (XVIII Hydrochloride) To a refluxing solution of XVI (214 g, 0.294 mole) and Ν,O-bis-(trimethylsilyl)acetamide (40 ml, 0.15 mole) in dry CK^Cl^ (2.9 L) was added dropwise, with stirring, a 50% solution of chloroacetaldehyde (93 g, 0.59 mole) in CHCl^ over a period οι 15 minutes. After standing for 30 minutes, the mixture was concentrated to dryness. To the residual oil were added CK-.C1-. (1.5 L), Girard Reagent T (99 g, 0.59 mole) and 10% aqueous HCl (300 ml), and the mixture was stirred hour at room temperature. The organic layer was washed with water (200 ml) and a saturated NaCl solution (200 ml), dried over MgSO., treated with 4 eated with 1 N HCl in CH^OH (300 ml) The filtrate was cooled to -10°C The mixture was charcoal (5 g) and filtered, an d 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 (53%) of the title compound XVIII as its hydrochloride, melting at >185°C (dec.). Pale yellow needles. Α -3 ·£ JL A IR : v * cm ‘ max 2830, 1780, 1720. UV : ,EtOH λ nm ma x (e) 286 (8800). NMR :6DHSO-ds 3.73 (2H, br, s, 2-H), 3.97 (2H, m ppm (IH, d, J-4.5 HZ, 6-H), 5.37 (IH, 7-H), 5.77 (IH, m, 3-CH»CH), 6.45 Hz, 3-CH), 6.88 (IH, S, CHPhj), 7. s, Ph-H).

, CH,C1), 5.22 d, J«4.5 Hz, (IH, d, J~ll (10H, br, Anal. Calc’d for C,3H2 , N^SCl HCl: C, 57.87,- H, 4.65,N, 5.87,- S, 6.7 2; Cl, 14.85.

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

Preparation No. 13 Banzhydryl 7-[2-(5-Amino-1,2,4-th ia diazol-3-yl)-2-methoxyiminoacetamido]-3~[3-chloro-l-propen-l-yl)-3-cephem-4-carboxylate ( 2 isomer ) (VUI-1) To a stirred solution of XVZII (Z isomer) (20 g, 42 mmole) in CHjCl, (420 ml) containing N,O-bis(trimethylsilyl)acetamide (34 ml, 125 mmole) was added 2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetyl chloride hydrochloride (15.2 g, 59 mmole) in three portions over a period of 30 minutes at -10 to 0°C. The mixture was stirred for 30 minutes at 0-5eC and concentrated under reduced pressure. The residual brown oil was dissolved in ethyl acetate (420 ml) and the solution was washed successively with saturated aqueous NaHCOj (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 28.5 g (90% pure) of the title compound Vin-l (Z-isomer) as a colorless powder. M.p. >150°C (dec.).

IR : vK9r cm'1 3400, 1780, 1720, 1680, 1620. max UV : xEt0Hnm (£) 240 (20000), 283 (12000). max NMR : 6acetone"d6 3.6 (2H, m, 2-H), 3.95 (3H, s, OCHg), 4.0 ppm (2H, m, CH^Cl), 5.32 (IH, d, J=4.5 Hz, 6-H), 5.62 (IH, m, 3-C0=CB), 6.03 (IH, d, J«4.5 Hz, 7-H), 6.32 (IK, d, J=ll Hs, 3-CH), 6.87 (IH, s, CHPhg), 7.33 (10H, br, s, Ph-H).

Preparation No. 14 Ben zhydryl 7-f 2~(5-Amino-l,2,4~thiadiazoI-3-yl)-2-methoxyiminoacetamido)-!- [3-iodo-l-propen-l-yl)-3-cephem-4-carboxylate (Ξ isomer) (IX-1) A mixture of VIII-1 (Z isomer) (28.5 g, 90% pure) and sodium iodide (19 g) in dry acetone (420 ml) was stirred for X0 minutes at room temperature and allowed to stand at 5®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 MgSO^ and evaporated to about 50 ml of volume. The concentrate was diluted with n-heptane (200 ml) to yield 30.6 g (95% pure) of the title compound IX-1 (E isomer) as a yellow powder, melting at >120eC (dec.).

KB f - Ί IR : υΛΟ'· cm * 3400, 1780, 1725, 1680, 1620. max uv : XEt0H nm (e) 306 (15000). max NMR : . acetone-el, ο Θ PPm 3.71 (2H, m, 2-H), 3.97 (3H, s, OCH,), 4.0 —3 (IH, d, JM.5 Hz, 6-H), 6.03 (IH, d-d, J<=4.5 & 8 Hz, changed to doublet J«4.5 Bz by D,O, 7-H), 6.32 (IB, d-t, 8 Hz, 3-CH-CH), 6.79 (IH, d, J-15 Hz, (2H, d, J«8 Hz, CH,!), 5 J«15 & 3-CH), 6.98 (IB, s , CHPh 2 ), 7.35 (10B, m, Ph-H), 7.63 (2H, be, s, disappeared by D^O, NH2), 8.52 (IH, d, J-8 Hz, disappeared by D,O, 7-NH).

Preparation No. 15 Ben zhydry17- [2-(5-Amino-1,2,4-th iadi azol-3-y1)-2-methoxyiminoacetamido 3-3-(3-(4-carbamoyl-l~pyridinio)-1-propen-l-yl]-3cephem-4-carboxylate Iodide (E isomer) (XII-1B) To a suspension of IX-1 (E isomer) (30.5 g) and isonicotinamide (26 g, 212 mmole) in CB^CN (120 ml) was added CHjOH (100 ml) until the mixture became clear. The solution was stirred for 2 hours under nitrogen atmosphere at room temperature and 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 and the residual yellow powder was washed with a mixture of isopropyl ether and CH^OH (3/1, 120 ml). The powder was collected by filtration and dried in vacuo to give 36 g (75% pure estimated by HPLC, of the title compound XI2-1H (E isomer) as a light yellow powder melting at τη >150*C (dec.).

JL A IR : vKBc cm'1 3300, 1780, 1720, 1680, 1620. max UV ·. X£t0Hnm (e1%H ) 282 (170). max 1cm N MR .DMSO-d, o o PPrc 3.72 (2H, m, 2-H), 3.90 ( 3H , s, OCH-j) , 5.25 (3H, m, 6-H & CH,N+), 5.9 (IH, d-d, J«4.5 ί 8 Hz, changed to a doublet J«4.5 Hz by D^O addition, 7-H), 6.35 (IH, m, CHPh?), 6.9 (IH, d m, Ph-H), 8.06 (2H 3-CH--CH), 6.89 (IH, S, J«16 Hz, 3-CH), 7.35 (10H, br, s, disappeared hy D?O, NH?), 8.21 (2H, addition , NH^), Py-H), 9.57 (IH br, s, disappeared by D?O 8.36 & 9.07 (each 2H, d, J«6 Hz, d, J=8 Hz, disappeared by D?O addition, 7-NH).

Preparation No. 16 Benzhydryl 7-Benzylideneamino-3-[3-chloro-I-propen-1-yl)-3cephem-4-carboxylate (XVII) (2 isomer) To an ice-cooled mixture of the crystalline 7-amino20 cephem intermediate XViu (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 1O*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 tic to confirm that all of the starting 7-amino4 li. JL cephalosporin was converted to the Schiff's base. Removing the solvent jn vacuo gave 16.45 g of the title compound XVII (z isomer) as a pale yellow powder (estimated purity 85%; M.p. 74°C (dec.), which was used for the next step without purification.

IR : KBr v cm max1 1780, 1725, 1635. UV : A z 2 nm (E1% ) 257 (400). max 1 cm NMR : gCDCig 6.18 (IH, d, J«ll Bz). 10 PP*- Preparation No. 17 Benzhydryl 7-3enzylideneamino-3-[3-(4-carbamoyl-l-pyridinio)-1propen-l-yl 3-3-cephen'.-4-carboxvlate Iodide (XXI-K) (£ isomer) To a chilled mixture of the 3-chloropropenylcephem. xvn (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 (E1 % (255 nmJ/E1 % (320 nm)]. 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 >5 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. To the combined solution of the filtrate and washings, was added a solution of isonicotinamide (3.5 g, 28.7 JO mmole) in dimethylformamide (20 ml) and the mixture was concentrated under reduced pressure. The concentrate was allowed to stand at room temperature for 1.5 hours and washed with isopropyl 118 ether (100 ml x 3). 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 L). 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 (£ isomer) was obtained. Yellow amorphous powder. M.p. 150-155°C (dec.,. Estimated purity 80% by nmr.

IR : vKBr cm1 1775, 1725, 1690, 1635. max UV : λ χ £ nm (E1% ) 258 (335 ) , 296 (255) • max 1 cm NKH ; .^DMSO-dg PP* 3.4-3.8 (2H, b r .), 5.35 (2K, br . ), 5 .41 ( IH , d J=4 Hz), 5.73 (IH, d, 4 Hz) , 6.93 (IH, s) 9 6.97 (IH, d, J=16 Hz), 7 . 3-7 . 5 (15H, br . s) 9 8.40 (2H, d, J~6.5 HZ) , 9.15 (2H, d, J«6 .5 HZ ) Preparation No. 18 7-Amino-3-[3-(4-carbamoyl-l-pyridinio)-1-propen-l-yl3-3-cephem~4carboxylate (XXII-H) (E isomer) To a suspension of the quaternized cephem XXI-H (17 g) in 85% 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 charcoal. The mixture was filtered and washed with 85% formic acid (5 ml). The filtrate was combined with the wash and poured into acetone (1 L), 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 (50 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 119 collected by filtration, washed with a small amount of methanol (30 ml), and dried to give 7.53 g of the title compound XXII-K (& isomer) as the hydrochloride. Light yellow powder. Estimated pu r i ty 85% by UV . M.p. 17 3- -188°C (dec.). :r : KBr Ί v cm 1795, 1680, 1620, 1575, 1540. max UV : ,Phosphate buffer (pH7> nm (E1 % , 294 (457). max 1 cm NHR :6D,O+DC1 ppn1· 3.82 (2H, s, , 5 . 17 (IH, d, J=5 HZ), d, J = 7 Hz), 5.43 (IH, d, J=5 Hz), 6.37 (IK, d-t, J=16 & 7 Hz), 7.23 (IH, d, J=16 Hz), 8.34 (2H, d, J = 7 Hs), 9.00 (2H, d, J=7 Hz).

Preparation No. 19 2-(5-Amino-1,_2,4~thiadiazol-3-yl) - 2-methoxy im.in pace tyl Chlor ide Hydrochloride (III-l _as its acid chloride hydrochloride) 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®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 sulfate (563 ml, 6 mole, at 15-20’C end 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 (77%) of the title compound as brown needles melting at 170-172®C. 0 rz ώ r — 1 IR : v cm " 3400, 3180, 1720(sh), 1715, 1690, max 1615, 1570.

UV : λΗ2° nm (ε) 238.S (8290), 268 (sh, 3870). max NMR: &DMS0~d6 4.20 (3H, s, OCH-j), 7.85 (2H, br. NH,). ppm Anal. Calc'd. for Ο.Η,Ν,Ο,: C, 37.80,· H, 3.97; N, 33.06 j «3 Found: C, 37.43; H, 3.75; N, 32.51.

B. 2-Methoxyiminopropenedini trile A stirred mixture of 2-cvano-2-methoxyiminoaeetamide (86.9 g, 0.7 mole), sodium chloride (70 g) and phosphorus oxychloride (97 ml, 1.05 mole) in dry 1,2-dichloroethane (350 ml) was refluxed for 16 hours. The insolubles were filtered off through a dicalite pad and washed with dichloroethane. The filtrate and the wash were combined, and poured into stirred ice-water (1.5 L) to decompose the excess of phosphorus oxychloride. The organic phase was washed with 10% NaHCO^ (500 ml), water (500 ml x 3) and a saturated NaCl solution (500 ml), and dried over MgSO„. The filtrate was distilled under «SB diminished pressure to give 61.5 g (81%) of the title compound boiling at 62°C/24 mm ag. (Lit., b.p. 47-48°C/12 mm Hg).

Liauid Film ma x cm * 3020, 2960, 2245, 2020, 1530, 1455, 1080.

NMR: έ> 3 ppm 4.35 (3H, s, OCH 3).

IR 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-methoxyiminopropanedxnitrile (58.0 g, 0.53 mole) in ethanol (120 ml) at -15 to ~10°C over a period of 30 minutes» with stirring. The mixture was stirred at -10eC overnight and then at ambient temperature (20-25*0 for one day. The reaction mixture was partitioned between water (350 ml) and CK^Cl? (350 ml), and the aqueous phase was saturated with sodium chloride, and extracted again with CH^Cl? (300 ml). The organic extracts were combined» dried over MgSO^ 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 67.5 g (69%). M.p. 152-4°C (dec.). [Lit , m.p. 150-155°C (dec.)].

IR : vKBr cm1 3160, 2900, 2360, 2235, 2000, 1665, 1555, 1495, 1415. (ε) 243 (8500), 265 (sh, 5380), 305 (sh, 1400). rv . ,EtOE Lv : a nm. max Mvro. -DHSO-d_, NMR: ο o ppm Anal. Calc'd for 1.88 (3H, s, CHCOOH), 4.15 (3H, s( OCH3), 7.60 (4H, br.).

C4HgN4O-CH3COOH: C, 38.71; Η» 5.41; Ν» 30.09 Found: C, 38.71; H, 5.59; Ν» 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 CH3OH (1.25 L) were added dropwise triethylamine (234 ml, 1.68 mole) at -10eC, and subsequently Br? θ'* (41.6 ml, 0.806 mole) over 20 minutes at -15 to ~10°C, and the mixture was stirred minutes ?o the mixture was added dropwise a solution of KSCN (78.3 g, 0.806 mole) in CH^OH (550 ml) over l hour at -15 to -10°C. After stirring at 0-5°C for 1 hour, the mixture was poured into ice-water (12 L) to form a crystalline precipitate, which was collected hy filtration, washed with water and air-dried to give 120 g (98%) of the title compound. M.p. 263-5°C (dec.). The m.p. of the compound prepared by us is higher by about 60"C than that given in the literature* [m.p. 210-15°C (dec.)), but our spectral and .be structure microanalytical data are consistent IR : KBr -1 υ cm 34 35, 3260, 3120 max 1545, 1455, 1415 UV ; XEfc0H nm ( ε ) 248 ( 13300), maxxDMSO-d, NMR: δ 0 ppm 4.21 (3H, s, OCH Anal. Calc’d for C5H5N5OS: C, 32.78; H, 2.75; N, 38.23; S, 17.50 Found Ct s. 32.76; H, 2.51; N, 38.02 17.50. 2-(5-Amino-l,2,4-thiadiazol-3-vl)-2-methoxviminoacetic Acid (1 •1) A mixture of 2-(5-amino-l,2,4-fchiadiazol-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^PO^, and washed with ethyl ml x 2, and 200 ml x 1) NaCl, and extracted three fc imes and tet rahydrofuran (3 ; 1, 300 tracts were combined , dried over uced pr essure. The residue was triturated with isopropyl ether to afford pale yellow crystals oj 3 the title acid. Yield 16.8 g (83¾) . M. p. 184-5’C (dec .) I bit m.p. 180-182*0 (dec. ) ] . KBr -1 IR : v cm 3460, 32 60, 3140, 1725, 16 20, 1605, 1545 . max UV : λΗ2° nm (ε) 234 (13200), 288 (sh, 3620). max 2-(5-Amino-l,2,4-th iadiazol-3-yl)-2-methoxyiminoacgtyl Chloride Hydrochloride To a suspension of 2-(5-amino-1,2,4-thiadiazol-3-yl)-2methoxyim.inoacetic acid (III—1) (40.4 g, 0.2 mole) in dry CH^Cl, (400 ml) was added PCl^ (41.6 g, 0.2 mole) in one portion at -50°C. The mixture was stirred for 4 hours at -20 to -5°C, and poured into a mixture of n-heptane and isopropyl ether (2 : 1, 2 L). The yellow precipitate was collected by filtration, washed with the same solvent mixture, and dried with KOH under reduced pressure to give 46.0 g (90%) of the title acid chloride.

XR NujOl -1 .-αν J cm 1 / 7s. max *Japan Kokai 57-158769 published September 30, (Brit, appl., 3/6/81) 1982, to Fujisawa 4 VIII-2 *2 Diphenylmethyl 7-(2-( 5-Amino-l, 2P_4-thiadig,sol3-yl)~2-(2)ethoxyiminoacetamido3-3-(3-chloro-l-propenyl3-3-cephem-4carboxylate (7111-2, Z isomer) ml To a mixture of N,O-bis(trimethyIsilyl)acetamide (2.3 mmoles) and crystalline diphenylmethyl 7-amino-3-(3chloro-l-(Z)-propen-l-yl]-3-cephem-4-carboxylate hydrochloride (XVIII) (1.338 g, 2.8 mmoles) (from Preparation No. 12.) in methylene chloride (10 ml) was added 2-( 5-amino-l, 2, 4thiadiazol-3-yi)-2-{Z)-ethoxyiminoacetyl chloride hydrochloride (800 mg, 2.95 mmoles) portionwise, with stirring, at -1Q°C and the mixture was allowed to stand at 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 (95%). Mp. >150°C (dec.).

IR v . (KSr) in cm ma x 3300, 1780, 1720, 1690, 1380, 1220 UV : , (C-H-OH) in nm (e) ma x 2 5 NMR : 6 (DMSO-d-) in Dorn 0 1.26 ( 2H, ( 3i 285 (11000).

J == 7Hz, CH,CH3), 4.25 , CH-jCHg), 5.90 (IH, 6.26 (1H, d, s, CHPh,), q ι J = 7 Η z d-d, J=4 & 8Hz, 7-H), J=llHz, 3-CH), 6.85 (IB, (IH, d, J=8Hz, 7-NH, P r epa ration No . 21 h2h C2B5 CONH CH-CH-CH.

COOCH(Ph) IX-2 *A mixture of E and 2 isomers Diphenylmethyl 7-(2-( 5-Amino-l, 2,4-thiadj.azol -3-yl) - 2-( 2) -ethox iminoacetam.ido ]-3-( 3-iodo-l-propenyl ]-3~caphem-4-carhoxylate {IX-2) mixture of Viil-2 (1.90 g, 3 mmoles) (; :om Preparation No. 20) and sodium iodide (1.4 g, 9 mmoles) in acetone (20 ml) was stirred for 10 minutes at room temperature and then allowed to stand at 5°C for 3 hours. The mixture was evaporated under reduced pressure, diluted with ethyl acetate (100 ml), washed with 10% sodium thiosulfate and water, and evaporated under reduced pressure. Trituration of the residue with isopropyl ether gave 1.82 g (84%) of the title product jxas a light brown amorphous powder. :r max (KBr) in cm 3290, 1770, 1720, 1670, 1530, 1370, 1220 .

UV λ (C_HeOH) in nm (E~J ) 304 (199). max 2 5 χ cjn .1% Ρ r e pa r a t i on No . 2 2 I COOCH (Ph), " \zx/ :0NH9 XXI-H iodide *S Diphenylmethyl 7~Benzylideneamino-3-f(£)-3-(4-carbamoy1pyridinio)-l-propenyl)-3-cephem-4-carboxylate (XXI-H iodide) (£ isomer ) To a chilled solution of the 3-chloropropenylc.ephem (XVII, Z isomer, 42.8 g- 90 mmoles) (from Preparation No. 16) in cry 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 of UV absorption [E* (255 nm)/ cm E1% (320 nm)]. When the ratio became below 1.10 (after 45 I cm minutes), the mixture was diluted with 800 ml of methylene chloride, treated with active carbon (4 g), and filtered. The filter cake was washed with 100 ml of CH2C1O. To the combined filtrate and washings was added isonicotinamide (14.64 g), and the mixture was concentrated under reduced pressure. The concentrate was kept at room temperature for 1.5 hours and washed with a mixture of toluene and n-heptane (1:1, 600 ml). The residual brown semi-solid was dissolved in CH^Cl, (100 ml) and the solution was added dropwise to ethyl acetate (3 L, with vigorous stirring. After drying over ?2°5 ΪΗ vacuo, 57.37 g (88%) of the quaternized title product XXI-H was obtained as the iodide. Yellow amorphous powder. Mp. ISO-ISS^C (dec.). This product was identical to that obtained by iodination with Nal (Preparation No. 17). Ί! Λ. ® Pre Da ration No . 2 3 HCl CH-CH-CH,-Cl XVI COOCH(Ph) Diphenylmethyl 7-Am.ino-3- ( 3-chloro-l-oropenyl)-3-cephem-4carboxylate hydrochloride (2 isomer) (XVIII, Hydrochlor ide) A 25% solution of chloroacetaldehyde (69 g, 0.22 mmoles) in CHCl^ was added to a solution of XVI (80 g, 0.11 mole in CHjCl? (1.1 t) containing N,O-bis(trimethyIsilyl)acetamide (16.2 ml, 0.06 mole) at -10*C in one portion, and the mixture wa allowed to stand overnight at 5°C, ihe mixture was concentrated to ca. 0.3 L, diluted with a mixed solvent of ethyl acetate and isopropyl ether (1/2, 0.6 L), treated with silica gel (Wakogel C-100, 60 g) and filtered through a dicalite pad. The filter cake was washed with the same solvent system (0.2 L). The combined filtrate and washing were concentrated to ca. 0.2 L, treated with Girard Reagent T (60 g, 0.26 mole) and 4N HCl (220 ml), and seeded with a few crystals of XVIII hydrochloride.

After stirring for 3 hours, the resulting crystals were collects by filtration, washed with water (0.5 L) and ethyl acetate (0.5 L) and dried in vacuo to give 37 g (70%) of the title compound xvm hydrochloride, melting at >185°C (dec.). Pale yellow needles. This product was identical to that obtained in Preparation No. 12. 128 Will *Z Diphenylmethyl 7-Amino-3-(3-chloro-l-propenyl)-3-cephem-4carboxylate hydrochloride (Z isomer) (XVIII, Hydrochloride) To a solution of chlor©acetaldehyde (25% solution in CHCl^, 628 mg, 2 mmoles) in CH?C1? (10 ml) were added N,O-bis(trimethylsilyl)acetamide (0.135 ml, 0.5 mmole) and XVl (728 mg, mmole), successively, at 5eC. The mixture was allowed to stand overnight at 5°C. The mixture was evaporated and diluted with a mixture of ethyl acetate and isopropyl ether (1/2, 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 (80%, of the title compound XVIII hydrochloride, melting at >185°C (dec.). Pale yellow needles. This product was identical to that obtained by Preparation No. 12.

Preparation No. 25 2-(5-Amino-l,2,4-thiadiazol-3-vl)-2-(pr open-3-vloxyimino)acetyl chloride hydrochloride (III-3 as its acid chloride hydrochloride A. Methyl 2-(5-t-butoxyca rbonylamino-1,2,4-thiadiazol-3-yl)2-(propen-3-yloxyi.m.ino)acetate A mixture of 685 mg (3.37 mmoles) of N-(propen-3yloxy[phthalimide (prepared according to the procedure of E. 9 Grochosaki & J. Jurczak, Synthesis 1976 682] and 175 mg (3.35 mmoles) of hydrazine hydrate in 5 ml of C^HgOH 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 2-(5-t-butoxvcarbonylamino-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/ethy1 acetate (4:1) and fractions containing the major product were combined and evaporated under reduced pressure. Yield 514 mg (46%). Mp. 83-86°C.

IR v υ (KBr) in cm ma x 3100, 1745, 1710, 1610 UV Xm.ax H5OH) in nm (e) 223 (5700), 242 ( 10000) NMR : 5 (CDC13) in ppm 1.55 ( 9H , S, BOC-H), 4.40 (2H, d, J=5Hz, O-CH2), 5.21 (2H, m, CH,=CH), 5.90 (IK, m, -CH=CH,), 9.50 (IK, br.s, NH ) . 2-(5-t-3utoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2(propen-3-yloxyimino)ac et i c aci d1 A solution of 770 mg (2.3 mmoles) of methyl 2-(5-tbu toxyca rbonylamino-1,2,4"thiadiazol"3-yl)~2"(propen-3-yloxyxmxno) acetate and 3.5 ml of 2N NaOH solution (7.0 mmoles) in 15 ml of CH~OH was refluxed for 30 minutes. The reaction mixture 3 was concentrated in vacuo and diluted with 10 ml of ethyl acetate-HjO (1:1). The water layer was separated, acidified to pH 2 with 6N HCl and extracted with ethyl acetate (10 ml x 2). The ethyl acetate solution was dried over MgS04 and concentrated by a rotary evaporator to afford 596 mg (81%) of the title compound , Mp. 134-135CC ( ,.fcl).

I f· · <«» A 9 mp. 135-136’C.

IR υ „ (Nujol) in cm j ltd Λ 3150, 1745, 1710, 1550 0 uv : λ^χ (C9H5OH) in nm (ε, 223 (11000 ) , 242 ( 11300 ).

NMR : δ (DMSO-dg) in ppm 1.55 ( 9H , s, BOC-H) , 4.77 (2H, d, J=5Hz, O-CHg), 5.22 (2H, m, CH7*CH>, 6.0 (IH, m, CH=CH,). 1) I. Csendes, et a1., J. Antibiotics, 36 , 1020 ( 1983).

C. 2-(5-Amino-l,2,4~thiadiazol-3-yl)-2-{propen-3-yloxyimino)acetic acid (III-3)11 A solution of 570 mg (1,74 mmoles) of 2-(5-t-butoxycarbonyl amino-1 , 2,4-thiadiazol-3-y1)-2-(oropen-3-yIoxyimino,acetic acid in 6 ml of trifluoroacetic acid was allowed to stand for 1 hour at ambient temperature. Evaporation followed by trituration with 30 ml of isopropyl ether gave 376 mg (95%) of. the title compound. Mp, 109°C (dec.).

XR : υ (Nujol, in cm"1 3180, 1710, 1545, 1460. ma x UV : (C h OH) in nm (ε) 245 (13500). ma x zo NMR : ό (DMSO-dg) in ppm 4.77 (2H, d, J=5Hz, O-CH9), 5.20 (2H, m, CH=CH), 6.0 (IH, m, CH=CH7). 1) Japan Kokai 57-112396 (7/13/82, Fujisawa) Brit. appl. 7935538 (10/12/79).

D. 2-(5-Amino-1,2,4-thiadiazol-3-y1)-2-(propen-3-yloxyimino)acetyl chloride hydrochloride A solution of 350 mg (1.54 mmoles) of III-3 and 410 mg (1,97 mmoles) of phosphorous pentachloride in dichloromethane (5 ml) was stirred for 1 hour at 25*C. The reaction mixture was poured into 60 ml of n-hexane and the precipitate was filtered off. Yield 323 mg.

IR : v (Nujol) in cm"1 1765. m «5 x 131 Pre oarati on 2-(5-Amino-1,2,4-thiadi chloride hydrochloride azol-3-yl)-2-propargyloxyiminoacetyl (III—4 as its acid chloride hydrochloride) A. Methyl 2-(5-1-Butoxycar bonylamino-l,2,4-thiadiazol-3-yl)5 2-propar gyloxyiminoacetate A suspension of 870 mg (4.32 mmoles) of N-propargyl1 ) oxyphthalimide and 200 mg (4.0 mmoles) of hydrazine hydrate in 5 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-c-bucoxycarbonylamino-l,2,4-thiadiazol-3-yl)-2oxoacetate"\ The solution was allowed to stand for 1 hour and concentrated under reduced pressure. Purification by silica gel chromatography followed by evaporation afforded 319 mg (27%) of_ the title product. Mp. 72-75’C. :r uv v (KBr) in cm " 3200, 2380, 1745, 1710, 1610 max λ _ (C-HcOH) in nm (e) 235 (12200) m.cs x 2 5 NMR : δ (DMSO-dg) in ppm 1.56 (9H, s, BOC-H), 3.55 (IH, t, J-2HZ, C==CH), 4.85 (2H, d, J~2Hz, -CH,-C=CH), 8.9 (IH, br.s, NH). 1) Commercially available, Aldrich. 2) I. Csendes et al., J. Antibiotics 36, 1020 (1983). 2-(5-1-Butoxycarbonylamino-1,2,4-thiad iazol-3-yl)- 2propargyloxyiminoacetic acid A solution of 490 mg (1.4 mmoles) of methyl 2-(5-t25 butoxyearbonylami no-l,2,4-thiadiazol-3-yl)-2-propargyloxyiminoacetate and 2.2 ml of 2H aqueous NaOH solution (4.4 mmoles) in 14 ml of CHjOH was refluxed for 30 minutes. The reaction mixture was concentrated under reduced pressure and 10 ml of ethyl i. ό **> acetate-H^O (1:1) was added to the solution. The separated water layer was acidified to pH 2 with 6N HCl and extracted with ethyl acetate (2 x 10 ml). Drying over MgSCL followed by evaporation · of the organic layer gave 149 mg (89%) of the title product. Mp. 135°C (dec.).

IR : v (Nujol) in cm ~ 3350, 1720, 1570, 1550. ΓΠ3 X UV S λ (C.HcOH) in nm (e) 233 (11500). ma x 2 5 NMR : ό (DMSO-dg) in ppm 1.55 ( 9H , s, BOC-H), 3.55 (IH, t, J=2Hz, C—CH) , 4.89 (2H, d, J*2Hz, CH2C=CK ), 9.0 {IH, s , NH ) .

C. 2-(5-Amino-i,2,4-thiadiazoi~3-yl)-S-propargyloxviminoacetic acid (III—4) A solution of 410 mg (1.26 mmoles) of 2~{5-t-butoxycarbonylamino-l,2,4-thiadiazol~3-yl)-2-propargyloxyiminoacetic acid in 5 ml of trifluoroacetic acid was allowed to stand for 1 hour at 25®C. Evaporation followed by trituration of the residue with 25 ml of isopropyl ether gave 204 mg (72%) of the title compound. Mp. 156-158eC (dec.).

IR : v (Nujol) in cm 1 3300, 2480, 1730, 1610. ΣΛ3 X uv : λ (C-H,OH) in nm (ε) 234 (12000). ma x a j NMR : 6 (DMSO-dg) in ppm 3.52 (IH, t, J=2Hz, CSCfl), 4.86 (2K, d, J=2Hz, CH2-C^CH), 8.10 (2H, br.s, NH0).

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

D. 2-(5-Ami no-1f 2,4-thiadiazol-3-yl) -2-propargy1oxyiminoacetyl 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 °C. The reaction mixture was poured into 30 ml of n-hexane and the precipitate was filtered off. Yield 65 mg (34%).

IR : = (Nujol) in cm-1 1770. max Pre pa r at i on No . 2 7 2- ( 5-Amino-l, 2, 4-thiadiazpl-3-yl)-2-cycl opentyloximinoe,cetyl chloride hydrochloride (II1-5 as its acid chloride hydrochloride) A. Methyl 2-(5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2cyclopentyIoxyiminoacetate A suspension of 860 mg (3.7 mmoles, of N-(cyclopentyloxy)phthalimide1) and 185 mg (3.7 mmoles) of hydrazine hydrate in 5 ml of was stirred for 1 hour at ambient temperature and filtered. The filtrate and washings were combined and added to 1.06 g (3.7 mmoles) of methyl 2-(5-fc?) butoxycarbonylami no-1,2,4-thiadiazol-3-yl)-2-oxoacefate . The solution was allowed to stand for 1 hour at room temperature and concentrated in vac up. 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 l 81%). Mp. U5~118°C. IR : υ max (KBr) in cm •1 3200, 1745, 1710, 1550. (JV : λ max (C,H5OH) in nm (e) 217 (1800) , 252 (7600).

NMR : ό (CDCip in ppm 1.51 (9H, s, BOC-H), 1.60 (8H, br.s, H~("] ), 3.88 ( 3H , s, OCH3), 4.90 (IE, br . s ), 8.70 (IH, br.s, NH) 1) U.S. Patent 3,971,778 (7/27/76; Glaxo), Brit. appl. 49255 (10/25/72). 2) I. Csendes et al., J. Antibiotics 36, 1020 (1983). 2-(5--1-But oxy car bony 1 ami no-1,2,4-thiadxazoI-3-yl)-2-cyclope nt yloxyiminoacetic acid A solution of 500 mg (1.34 mmoles) of methyl 2-(5-tbutoxyearbonylamino-1,2,4-thiadiazol~3-y1)-2-cyclopentyloxyiminoacetate and 2N NaOH solution (2 ml, 4 mmoles) in 15 ml of CH,OH was refluxed for 30 minutes. The reaction mixture was evaporated and 10 ml of ethyl acetate-Η,Ο (1:1) 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 2). The organic lever was washed with brine, dried over Me SO, and concentrated under reduced pressure to give 377 mg (78%) of the title compound. Mp. 185°C (dec.).

IR : v (KBr) in cm ~ 3160, 1710, 1550, RleX UV : λ (C.HcOH) in nm (ε) 238 (13300) ma x 2 5 NMR ; δ (DMSO) in ppm 1.51 (9H, s, BOC-H), 1.70 (8H, br.s., } ι 4-®~{iBt’ X3 O' C. 2-(5-Amino-l,2,4-thiadiazol3-yl)-2-cvclooentvloxvimino3) acetic acid (III-5, 2 isomer)' A solution of 348 mg (0.97 mmoles) of 2~(5-t-butoxyca rbonylamino-1,2,4-thiadia2ol-3-yl)-2-cyclopentyloxyiminoacetic Φ O’ κ J. Hi* 5ft 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 (86%) of the title compound. Mp. 162-165°C (dec.) (lit1); mp. 1SO-165°C (dec . ) ].

IR : V max (Nujol, in cm1 3290, 3200, 1710, 1615, 1600. UV : λ max (C?H5OH) in nm (e) 238 (13300). NMR : ό (DMSO-d5) in ppm 1.17-2.10 (8H, m, , 4.60-4.98 (IH, m), 8.22 (2H, s). 3) Japan Kokai 57-158769 (9/30/82, Fujisawa) Brit. appl. 8107134 (3/6/81).

D. 2- (5-Am.ino-l, 2,4-thiadiazol-3-yl)-2-cyclopentvloxyiminoacetyl chloride hydrochloride A solution of 190 mg (0.74 mmole, of 111-5 and 219 mg (1.0 mmole, of phosphorous pentachloride in dichloromethane (5 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 hy filtration. Yield 122 mg (60%).

IR : vmax (Nujol) in cm1 1760.

Preparation No. 28 Benzotriazol-1-yl-2-(5-amino-l» 2,4-thiadiazol-3-yl)-2-methoxyiminoacetate A mixture of 1-hydroxyhenzotriazole (2.7 g, 20 mmoles) and dicyclohexylcarbodiimide (4.12 g, 20 mmoles) in 65 ml of DMF was stirred at room temperature. After 15 minutes, III-l (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 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 (82%) of the title compound as a. light grey powder. Mp. 189-192°C (dec.) IR v (KBr, in cm ~ 1815, 1620, 1540, 1415, 1090, 1060, max 1005, 945, 865, 740.

Claims

1. ) 7-(2-(5-amino-l,2,4-thiadiasol-3-y1)-2-methoxyiminoacetarnido]3-[3-( triae thylammonio)-l-propen-l-yl 3-3-cephem-4-carboxylate,

2. ) 7-(2-( 5-amino-l s 2 f 4-thiadiazol-3-yl)-2-mefchoxyiminoacetamido]3-03-( 1-methylpyrrolidinio)-l-propen-l-yl3-3-cephem-4carboxylate, 2 “ wherein R is the same as R~ or is a group of the formula s in which X, R and R are as defined above,,, 3 is a conventional carboxyl-protecting group, 3 is hydrogen or a conventional amino-protecting group, 2 ie chloro, bromo or iodo, 3. -£3-(4-carboxymethylthiopyridinio)-l-propen~l-yl3-3-cephe»-4 carboxylate. 29. A pharmaceutical composition which comprises 3) 7-[2-( 5-amino-l,2,4-thiadiasol-3-yl)-2-methoxyiininoacetamido]3-(3-pvridinio“l-propen-l-yl]-3-cephea-4-carboxylate„ 3. •COOS* 147

3. The compounds of claims 1 or 2 wherein R is (lower )alkyl, cvcloalkvl of 3 to 5 carbon atoms, 1-carboxycycloalk-l-yl of 3 to 5 carbon atoms, allyl, propargyl or carboxy (lower)alkyl. 4. ) 7-(2-( 5-amino-l, 2»4-thiadiazol-3-y1)-2~methoxyiminoacetamidoj3-(3-( 3-aminopyr idinio) -l-propen~l-yl]-3-cephea-4-carboxylate e 4 5 1 in which X, R and R are as defined'above, 3“ is a con2 ventional carboxyl-protecting group and 3 is hydrogen or a conventional amino-protecting group. 2θ The process of clain 26 , whereby compounds 1-22 are aanufactursd,

4. 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetarnido)-3-(3-(tr imethylammonio)-1-propen-l-yl)-3~cephem-4~carboxyiate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 4 u hydrogen, methyl or ethyl, or R and R J , taken together with the carbon atom to which they are attached, may be a cycloalkylidene ring containing from 3 to 5 carbon atoms, and □ md R are each independently .5 :Q is a quaternary ammonio group,· or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 5. An effective antibacterial amount of at least one compound of formula I, or a salt, hydrate or solvate thereof, plus a pharmaceutically acceptable carrier. 30. Medication comprising an effective antibacterial amount of at least one compound of formula I, 5) 7-(2-( 5-amino-l,2,4-thiadiasol-3-y1)-2-methoxyiminoacetamido]3-03-( 3-foraylasainopyr idinio)-l-propen-l-yl ]-3-cephem-4carboxvlate, 5. 4 4 is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolysable esters thereof, which process comprises reacting a compound of the formula b 2 hn CH«CHCs^ 5 R is hydrogen, a straight-br branched chain alkyl group containing from X to 4 carbon atoms,, a cycloalkyl or cycloalkenyl ring containing fro» z 3 to 6 carbon atoms, or a group of the formula *5

5. 7-(2-(5-amino-l, 2,4thiadiazol-3-yl)-2-methoxyiminoacetamido1-3-(3-(1-methylpyrrolidinio,-1-propen-l-ylJ-3~cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 6. ) 7-(2-( 5-amino-l„2,4-thiadiasol-3-y1ί-2-methoxyiminoacetamido]3-(3-( 3~aainomethylpyr idinio )-l-propen-X-yX3~3-cephem—4carboxylate,

6. 7-(2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamidoJ-3-ί 3-pyridinio-lpropen-l-yl ]-3-cephem-4~carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 7. ) 7-(2-( 5-amino-l,, 2,4-thiadiaaol-3-yl)-2-aefchoxyiminoacetaajido33-(3-( 3-carbaaoylpyridinio5-l-propen-l-yl3-3-cephem-4carboxylate, 7-(2-(5-amino-1,2, methoxyiminoacecamido]-3-(3-(2“amino-5j. 4 1 thiazolo( 4,5-c ] pyr id in io) -1-pr open-1-yl J- 3-ceph sip- 4-ca rboxyl ate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 7-(2-(5-amino-l,2,4tamidoI-3-ί3-(4-carbamoyl-4-csrhoxylate, or a nontoxic solvate, hydrate or thereof.

7. 7-(2-(5-amino-1,2,4thiadrazol-3-yl)-2-methoxyiminoace tarnido1-3-(3-(3-aminopyrid inio)-1-propen-1-yl]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 8. ) 7-(2-( 5-amino-l 2,4- thiadiasol-3-yl) -2-nethoxy iminoace tamido ]3-(3-( 4-carhamoyIpyridinio ) -l-propen-l-yl 3-3-eephes-4carboxylate,

8. 7-(2-(5-amino-l,2,4thiadi azol-3-v1)-2-methoxyiminoacetamido J-3-1 3-(3-formylaminopvridinio)-1-propen-l-yl]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 9. ) 7-(2-( 5-amino-l, 2,4-thiad£asol-3-yl)-2-aethoxyiminoacetamido]3-( 3-(2-me thyl thiazolio)-l-propen-l-yl ]-3-cephea-4-carboxy late,

9. 7-(2-(5-amino-l,2,4thiadiazol-3-y1)-2-methoxyiminoacetamido)-3-(3-(3-aminomethy1pyridinio)-1-propen-l-yl]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 10. Or a salt, hydrate or solvate thereof, in a pharmaceutically acceptable carrier, for use in a method of treating bactex'ial infections in a warmblooded mammal (including man) in need thereof. 31. The process of claim 26» substantially as 10) 7-(2-( 5-amino-l, 2»4-thiadiaseol~3~yl)-2-Esethoxyiminoacetamido]3-(3-(2-amino~5-thiazolo[4„5~c]pyridinio)-l-propen-l-yl]-3cephem-4-carboxylate, 10 hydroxy or (lower)alkoxy, and R' : and R are each independently 4 5 hydrogen, methyl or ethyl, or R and R, taken together with the carbon atom to which they are attached, say be a cycloalkylidene ring containing from 3 to S carbon atoms, and is a quaternary ammonio group, and nontoxic pharmaceutically acceptable salts and physiologically hydrolysable esters thereof, which process comprises reacting a compound of the formula wi th an acid of the formula B SN £>-COOH S3 \ _2* OR or with an acylating derivative of said acid.» wherein R* 2 same as R or is a grouo of the formula iis the ΐ COOB* 10 and » is aero or one, with a tertiary amine CSH (or sequentially with a secondary amine and a compound of the formula 51 - Ζ), and, if a ia 1, reducing the sulfoxide by conventional aeans, and subsequently removing all blocking groups by conventional meane. 27. A process for the preparation of compounds of the formula wherein ?» is hydrogen or a conventional aauno-protectmg group, R is hydrogen, a straight or branched chain alkyl group con5 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 in which R ie hydrogen, (lower)alkyl or carboxyl, X is halogen, 10 in which R” is hydrogen, (lower)alkyl or carboxyl, X is halogen, 4 4 hydroxy or (lower5alkoxy, and R and R” are each independently 4 3 hydrogen, methyl or ethyl, or R and R , taken together with the carbon atom to which they are attached, may be a cyclo™ alkylidene ring containing from 3 to 5 carbon atoms, and 10. 7-(2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido J-3-13-(4-{N-methy1carbamoyl)pyridinio)-1-propen-l-ylJ-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

10. 7-(2-(5-amino-l,2,4thi adiazol-3-y1)-2-methoxyiminoacetamido]-3-[3-(3-carbamoylpyridinio)-1-propen-l-yl]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 10 1° the claims which now follow,, the word ’’solvate” is to be understood as not including a ’’hydrate”. 137 CLAIMS R X 3N © : H«C H-C n 2 - N ” Q wherein R* is hydrogen cr a conventional amino-protecting group, 2 j------ _ ““-aight or branched chain alkyl group contain· a s i carbon atoms, a cycloalkyl or cycloalkenyl ring 3 to 6 carbon atoms, or a group of the formula R~ is hydrogen, ing from 1 to 4 containing from R 1 3 -C-CH=CH-R , 1 5 ; R •C-C3C-R' • 5 R 3 COOH or R 4 I •C-COOH I 5 R in which R is hydrogen, (lower)alkyl or carboxyl, χ is halogen, hydroxy or (lower)alkoxy, and R

11. ) 7-(2-(5-aaino-l, 2,, 4-thiadiatol-3-yI) -2-methoxyiainoacetamido]3-( 3-(4-hydroxyxaethylpyr idinio )-l-propen-l~yl3-3-cephem-4carboxylate,, 11. thiadiazol-3-v1,-2-methoxyiminoace pyridinio)-1-propen-l-yl]~3™cephem pharmaceutically acceptable salt, physiologically hydrolyzable ester 12. ) 7-(2-(5-amino-l,2 < 4-thiadiatol-3-yl)-2-methoxyiainoacetamido33-(3-(3-hydroxy»ethyIpyridinio)-1-propen-l-yl3-3-cephe«-4carboxylate,

12. 7-(2-(5-amino~l,2,4~ thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-1 3-(2-methylthiazolio,-1-propen-l-ylJ-3-cephem-4-carboxvlate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

13. ) 7-(2-( 5-amino-l, 2,,, 4-thiadiazol-3-y 1) -2-ae thoxy iminoace tamido ]3- (3-(4- K-aethylcarbaaoy1 pyridinio)-!-propen-1 - y 13 - 3 - c ephe m4- carboxylate, 13 . thiadiazol-3-yl)-2 13 8 A compound of Claim 1 selec ted from R 13 I N-R R 15 i s wherein R 1j , R 1- * and R 1 ^ are the same or different and ate {lowerJalkyl, {lower)alkenyl, &mino(lowerJalkyl with the provision that the amino may not be on an α-carbon, or hydroxy(lowerJalkyl with the provision that the hydroxy group may not be on an s-carbon; R 16 is hydrogen, (lowerJalkyl, {lowerJalkoxv, (lower)alkvifchio, amino, (lowerJalkylamino, di (lower ) alkyiamino, formylamino, (lowerJalkanoylamino, carboxy, hydroxy, carboxy (lower)alkyl, carboxy(lower )alkylthio, hydroxy(lower ) alkyl, halo(lowerJalkyl, aminoilowerJalkyl, (lowerJalkoxv(lowerJalkyl, carbamoyl or N-(lowerJalkylcarbamoyl, or R 1 ^ may represent a divalent alkylene group having 3 to 5 carbon atoms; R“ is (lower ) alkyl, (lower ) alkoxy (lowerJalkyl, halo(lowerJalkyl, allyl, hydroxy(lowerJalkyl with the provision that the hydroxy group is not on the e-carbon, amino(lower)alkyl with the provision that the amino group is not on the c-carbon, or phenyl (lower ) alkyl; R is hydrogen, (lowerJalkyl, (lowerJalkoxy, (lower)alkoxy(lower)alkyl, (lower ) alkylthio, amino, (lowerJalkylamino, di (lowerJalkylamino, carboxy, hydroxy,'carboxy (lower)alkyl, hycroxyilowerJalkyl, aminotlower ) alky 1, formylamino, (lower )alkanoylamino, carbamoyl or N-(lower )alkylcarbamoyl; n is an integer of from 1 to 3, inclusive; Z is CH. or, when n is 2, Z also may be S, 0 or N-R , 14. ) 7-(2-( 5-amino-l, 2,4-thiadiazol-3-yl)-2~iaethoxyiainoacetamido33-(3-(2,3-propylenspyridinio)-1-propen-l-yl3-3-cephem-4earboxylate,

14. 7-(2-(5~amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-(3-(4-hydroxymethylpyridinio)-1-propen-l-yl]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 15. Described in any of the foregoing Examples. 32. The process of claim 27» substantially as described in any of the foregoing Examples. 33. A compound of claim 1, prepared by a process as claimed in claim 26, 27, 28, 31 or 32. 15) 7-(2-(5-aaino-l,2,4-thiadiasol-3-yl)2-ethoxyiminoacetamido3— 3-(3-(4-carbamoyIpyr id ini o)-1-propen-1-yl3-3-cephem-4carboxylate,

15. 7-[2-(5~amino-l,2,4~ thiadiazol-3-yl)-2-methoxyiminoacetamido1-3-(3-(3-hydr oxymethvlpyridinio)-1-propen-l-ylJ-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

16. ) 7-(2-(S-amino-I,2, 4-thiadiazol-3-yl)-2-cyclopenfcyloxyijainoacefcamido]-3-(3-i4-carbaaoylpyridinio)-1-propen-l-yl3-3csphem-4-carboxylate, 17. ) 7-(2-( 5-amino-l, 2,4-thiadi azol-3-yl) -2-allyloxyisninoacetamido3-3-(3-(4-carbamoylpyridinlo)-I-propen~i~yl3“3cephem-4-carboxylate, IB) 7-(2-(5-amino-l,2,4~thiadi&sol~3-yl)~2~propargyloxyiminoacetaaido3-3-(3-(4-carbaaoyIpyridinio)-1-propen-l-yl3“3“ cephem-4-carboxyIate,

17. 7-[2-(5-amino-l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido .1-3- ( 3-( 2,3-propylenepvridinio)-1-propen-l-ylJ-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

18. 7-(2-(5-amino-l,2,4thiadiazol-3-yl,-2-ethoxyiminoacetamido 1-3-[3-(4-carbamoy1pyridinio)-l-propen-l-yl]-3-cephem-4-carboxylate f or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 19. ) 7-(2-(5-aaino-l # 2,4-thia

19. 7-[2-(5-amino-l,2,4thiadiazol-3-yl)~2-cyclopentyloxyiminoacetamido1-3-(3-(4carbamoylpyr idinio)-l-propen-l-yl )-3-cephem-4-carboxylate,· or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 19 z m which R is hydrogen or (lower)alkyl; and Η 2 θ and R 21 are the same or different and are hydrogen, (lower )alkyl, (lower ) alkoxy, (lower )alkylthio, amino, (lower )alkylamino, di (lower )alkylamino, carboxy, hydroxy, hydroxydower ) alkyl, amino(lower ) alkyl, (lower ) alkoxy (lower ) alky 1, carboxy (lower Jalkyl, carboxy(lower )alkylamino, (lower,alkanoylamino, carboxy(lower )alkanoylamino, carbamoyl or N-(lower ) alkylcarbamoyl . 20. 34. a pharmaceutical composition which comprises a compound as claimed in any of claims 1 to 25, or claim 33. plus a pharmaceutically acceptable carrier or excipient. 35. Use of a compound as claimed in any one of Claims 20) 7-(2-( S-asln.o-1,2,4-fchiadiasol-3-yl)-2-ethoxyiminoacetamidoj3-(3-(4-oarboxypyridinio)-2-propen-l-yl]-3~eeph«ffi-4carboxylate,

20. 7~[2-(5-smino-l,2,4thiadiazol-3-yl)-2-allyioxyiminoacetamido}-3-[3-(4-carbamoylpvridinio)-1-propen-l-yl3-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 21. ) 7-(2-(5-asino-l,2,4-fchiadia;coI-3-yl)-2-iaethoxyiainoacetamido3~ 3-(3-(4-carboxymethyIpvridinio)-1-propea-l-yl3-3-cephea-4carboxylate, and

21. 7-(2~(5-amino-l,2,4thiadiazol-3-yl)-2-propargyloxyiminoacetarnido3-3-(3-(4carbamovlpyridinioj-l-propen-l-yl]-3-cephem-4-carboxylate„ or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 22. ) 7-t2-(5-aaino-l,2,4-thiadiaxol-3-yl)-2-methoxyiminoacetamido]

22. 7-[2-(5-amino-l,2,4~ thiadiazol-3-yl)-2-methoxyiminoacetamido1-3-(3-(4-carboxypvridin.io)-1-propen-l-yl ]-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

23. 7-[2-(5-amino-l,2»4thiadiazol-3-yl)-2-ethoxyiminoacetamido1-3-(3-(4-carboxypyridinio)-2-propen-l-ylJ-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof.

24. 7-( 2-( 5-amino-l, 2» 4thiadiazol-3-yl)-2-methoxyiminoacetamido1-3-(3-(3-carboxymethyl pyridinio)-l-propen-l-ylJ-3-cephem-4-carboxylate, or a nontoxic pharmaceutically acceptable salt, solvate, hydrate or physiologically hydrolyzable ester thereof. 25. 7-(2-(5-amino~l,2,4thiadiazol-3-yl)-2-methoxyiminoacetamido3-3-(3~(4-ca rboxymethyithrcpyridinio)-1-propen-l-yl]-3-cephem-4-carboxylats, or a noncoxie pharmaceutically acceptable salt, solvate, hydrate or ίϊ 4 3 physiologically hydrolyzable ester thereof. 25. A process for the preparation of compounds of the formula wherein St 1 is hydrogen or a conventional aaino-protectmg group,

25. I to 25 for treating bacterial infections in a warm-blooded mammal (including man).