FI84830C - FOERFARANDE FOER FRAMSTAELLNING AV THERAPEUTIC ACTIVATE KEY - Google Patents

Description

1 84830

A process for the preparation of therapeutically active cephem derivatives

The invention relates to the process 7- [2- (5-amino-1,2,4-thia-5-diazol-3-yl) -2- (substituted) iminoacetamido] -3- [3-quaternary ammonio) -1- for the preparation of propen-1-yl] -3-cephem-4-carboxylates of the formula Nn-C CONH-i R ^ S \ 10 li iline

RHlN s' N \ 2 0 CH = CH-CH2®N = Q

0R cocP ^ 1 ^ 15 wherein R1 and R2 are as defined below and - ^ = Q is a quaternary ammonium group as defined below, and for the preparation of their salts and esters. The invention also relates to intermediate compounds for use in the process.

20 U.S. Patent 4,390,534 (issued June 28, 2006).

on 1983; Tsutomo Teraji et al. ) discloses cephem and cepham compounds of formula 25 R1-ff-C-CONHT-R3 VI J-K4 ^ 2 5

OR R

Wherein R 1 is amino or protected amino; R 2 is hydrogen, acyl, -J ": optionally substituted aryl, substituted alkyl, alkenyl, alkynyl, optionally substituted cycloalkyl, cycloalkenyl, or an oxygen or sulfur-containing 5-35 membered heterocyclic ring substituted with oxo-2,84830 oxo groups (oxo groups); R 3 is hydrogen or alkyl, R 4 is hydrogen, acyloxyalkyl, acylthioalkyl, optionally substituted pyridinoalkyl, optionally substituted heterocyclylthioalkyl, alkyl, halogen, hydroxy or optionally substituted thiazoloalkyl, and R 5 is carboxy or protected carboxy; COO 'if R 4 is optionally substituted pyridinoalkyl or optionally substituted thiazoloalkyl, and the dashed line indicates either a single bond or a double bond.10 European Patent Application 13,762 (published August 6, 1980) corresponds to the aforementioned U.S. Patent.

U.S. Patents 4,381,299 (issued April 26, 1983), 4,331,665 (issued May 25, 1982), and 4,332,798 (issued June 1, 1982) are each published in U.S. Patent 4,390,534. and are similar in content.

European Patent Application 62,321 (published October 23, 1982) discloses cephem compounds of the formula R 1 wherein R 1 is amino or a protected amino; R 2 is an optionally substituted lower aliphatic hydrocarbon group or cycloalkenyl; and the group of the formula "-N © '\' 384830 is an optionally substituted heterocyclic cationic group having more than one nitrogen atom; and pharmaceutically acceptable salts thereof. Intermediate compounds of formula 5 are also provided.

I 0 | CH -N φ I

* 2 * 4 λ \ /

10 OR R

wherein R1 and R2 are as defined above, R4 is a protected carboxyl group and xP is an acid residue.

European Patent Application 74,653 (published March 23, 1983) discloses cephem compounds of the formula r1- / ~ Vc-c ° nhi-j ^ Svi 3

vh JXX

? Wherein R 1 is amino or a protected amino; R 2 is an optionally substituted lower aliphatic hydrocarbon group, cyclo- (lower) alkyl or cyclo (lower) alkenyl; R 3 is (lower) alkylamino, N -protected (lower) alkylamino, di (Alem-30 pi) alkylamino, sulfo (lower) alkylamino, hydroxy (lower) alkylamino, N-protected hydroxy (lower) alkylamino, acyloxy (lower) alkyl, (lower) alkoxy (lower) alkoxy (lower) alkyl, di (lower) alkylamino (lower) alkyl, (lower) alkylthio (lower) alkyl, (lower) alkylthio, 35 (lower) alkoxy, (lower) alkoxy (lower) alkoxy, hydroxy- 84330 (lower) alkoxy, acyl (lower) alkyl, hydroxy (lower) alkylthio, di (lower) alkylamino (lower) alkylthio, nitrogen-containing unsaturated 5-membered heterocyclic group, nitrogen-containing unsaturated 5-membered hetero-5-cyclylthio, or nitrogen-containing unsaturated 5- or 6-membered heterocyclylthio (lower) alkyl which may be substituted with a suitable substituent (s); and R 4 is hydrogen or (lower) alkyl; or a salt thereof.

U.S. Patent 4,332,800 (issued June 1, 1982; Tsutomo Teraji et al.) Discloses, e.g. compounds of formula OR 2 CO 0 e wherein R 1 is amino or protected amino; R 2 is (lower) alkyl and X is hydrogen or carbamoyl.

European Patent Application 47,977 (published March 24, 1982) discloses cephem compounds having the formula '- 25 (O) A m

Am-T-C-CONH -.- V.

0-R, COcP

- 30 2 wherein m is 0 or 1; Am is optionally substituted amino; T is a thiadiazolyl group (attached to other groups via its two carbon atoms); R 2 is hydrogen, optionally substituted alkyl, cycloalkyl or optionally substituted carbamoyl; and R 1 is an optionally substituted thiazolio, an optionally substituted pyrazolo, a tri (lower) alkylammonium, or a pyridinyl group of the formula

Ra 5

Rc / ^ = ^ Rb wherein R * is substituted (lower) alkyl, wherein the substituent is cycloalkyl, phenyl, hydroxy, alkoxy, halogen, cyano, carbamoyl, carboxyl or sulfo, (lower) alkenyl or carboxy-substituted (lower) alkenyl , (lower) alkylthio or carboxy-substituted (lower) alkylthio, amino or monosubstituted amino, wherein the substituent is (lower) alkyl, (lower) alkanoyl or aminobenzenesulfonyl, di (lower) alkylamino, substituted carbamoyl, wherein the substituent is (lower) alkyl, hydroxy (lower) alkyl, (lower) alkoxy, hydroxy or cyano, di (lower) alkylcarbamoyl, thiocarbamoyl, cycloalkyl, phenyl, hydroxy, (lower) alkoxy, halogen, (lower) alkoxycarbonyl, (lower) alkanoyloxy, (lower) alkanoyl, carboxyl, sulfo, cyano, nitro or hydroxysulfo (lower) alkyl; Rb is hydrogen or carbamoyl, or has the same meaning as Ra; and Rc is hydrogen or has the same meaning as Ra; and their salts.

European Patent Application 25,017 (published March 11, 1981) is similar in content, although not formally related to the former.

European Patent Application 30,630 (published June 24, 1981) discloses 3-vinyl glycemic compounds of the formula R 1 -A-CONHn-v.

- - 35 CT CH * CH2 R2 6 84830 wherein R1 is an optionally protected amino-substituted heterocyclic group which may also contain halogen or a group of the formula

5 R3SO2HN

<Q- wherein R3 is (lower) alkyl; R 2 is carboxy or protected carboxy; and A is lower alkylene which may be substituted with amino, protected amino, hydroxy, oxy and a group of the formula = Nn / OR 4 wherein R 4 is hydrogen, cyclo (lower) alkenyl, (lower) alkynyl, (lower) ) alkenyl optionally substituted with carboxy or protected carboxy, (lower) alkyl optionally substituted with one or more carboxy, protected carboxy, amino, protected amino, cyano, phosphono, protected phosphono- and a heterocyclic group which itself may have substituents; and their salts.

This patent application relates in particular to compounds of the formula V 25 N - r - C-CONH -, - r ^ S ^

ΓΊΓ »Π JL

H2N ^ s ^ Ns o N'Vj ^ sCH »CH2 OR4 *

COOH

Wherein OR4 is methoxy, carboxymethoxy, tert-butoxycarbonylmethoxy or 1-tert-butoxycarbonylethoxy.

GB Patent Publication 1,399,086 (published June 25-75, 1975) generally discloses a very large number of 7,84830 cephalosporins of the formula R-C-CO-NH-j-S B \ ss Jy., ORa.

COOH

wherein R is hydrogen or an organic group, R "is an ether-forming monovalent organic group bonded through an oxygen carbon atom, B is> S or> S-> 0, and P is an organic group. In one embodiment, P may be e.g. a vinyl group of formula / R3

15 -CH = C

V

wherein R 3 and R 4 may independently be hydrogen, nitrile, (lower) alkoxycarbonyl or a substituted or unsubstituted aliphatic, cycloaliphatic, araliphatic or aromatic group. However, the 5-amino-1,2,4-thiadiazol-3-yl group is not identified as a possible R substituent and does not suggest or suggest that P may be a quaternary ammonio-substituted propenyl group.

U.S. Patent 3,971,778 and its patents 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 are similar in content.

30 European Patent Application 88,385 (published September 14, 1983) discloses compounds of the formula

R-C-CONH-I-R-SN

: 35 N

1r2 r4 8 84830 wherein R1 is (unsubstituted) thiadiazolyl; R 2 is carboxy (lower) alkyl or protected carboxy (lower) alkyl; R 3 is hydrogen, halogen or (lower) alkenyl; and R 4 is carboxy or protected carboxy. Although 1-propenyl is mentioned as one of the possible meanings of R3, only compounds in which R3 is hydrogen, chlorine or vinyl are described in the application as an example.

U.S. Patent 4,307,233 (issued December 22, 1981; to Daniel Farge et al.) Discloses, e.g. 3-vinyl-10 likephalosporin derivatives of the formula N-n— c - CONH -, - Ss \ I Γ il E? 1 / 1S B2nA's '' «O ^ -N C" -C »-n (

15 or5 I

COOH

Where R5 can e.g. be alkyl, vinyl, cyanomethyl or a protecting group such as 2-methoxyprop-2-yl, and R3 and R4 are alkyl groups (optionally substituted with hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl groups, or R3 and R 4 together with the nitrogen atom to which they are attached may form a saturated heterocyclic ring having 5 or 6 members, optionally substituted with another heteroatom of N, O and S, and optionally substituted with an alkyl group. The compounds are useful as intermediates in the preparation of 3-thienylcephalosporin derivatives. As Sii-30, it is not shown or suggested that the 2-aminothiazol-4-yl substituent be replaced by a 5-amino-1,2,4-thiadiazol-3-yl group or a quaternary ammonio-substituted propenyl group as the 3-substituent.

Published GB Patent Application 2,051,062 corresponds to these 35 U.S. publications and is similar in content.

9,84830

European Patent Application 53,537 (published June 9, 1982) discloses e.g. 3-vinyl-cephalosporin derivatives of the formula

5 N --- C-CONH -, - r ^ S "S

hAXn 0XXAch = cH- <3 OC-COOR 5 <i00R2 * 4 R3 Rb R 5 R 5 10 wherein R% and Rb5 are the same or different and represent hydrogen or alkyl or together form an alkylene group having 2 or 3 carbon atoms, Rc5 is an acid protecting group, R2 is an acid protecting group such as an ester, R3 and R4 are the same or different and represent hydrogen, alkyl (optionally substituted by hydroxy, alkoxy, amino, alkylamino or dialkylamino) or phenyl, or R3 and R 4 together with the nitrogen to which they are attached may form a saturated heterocyclic ring having 5 or 6 members, optionally having another heteroatom of N, O and S, and optionally having an alkyl group as a substituent. The compounds are useful as intermediates in the preparation of 3-thiovinyl cephalosporin derivatives. It does not suggest or suggest that the 2-amino-25-thiazol-4-yl substituent be replaced by a 5-amino-1,2,4-thiadiazol-3-yl group or a quaternary ammonio-substituted propenyl group as the 3-substituent.

U.S. Patent No. 4,423,214 corresponds to this application and is similar in content.

30 European patent application 53 074 (published on 2 June 1982) generally discloses a very large number of 3-vinyl cephalosporin derivatives having:

formula (OI

φ X1:;> 35 R ° la-NH- | -f S R °

CH = C-R J

C00R ° 2a 10 84830 wherein R ° la (in one of several embodiments) may be N —- r— C — CO- i T i h9n ^ sx n

5 ‘N

0 or5 where R5 can be e.g. hydrogen, alkyl, vinyl, cyanomethyl, an oxime protecting group such as trityl, etc., or a group of formula 10 -C-COORc5 R% / VRbs wherein Ra5 and Rb5 are the same or different and may be hydrogen, alkyl or together an alkylene radical having 2 15 or 3 carbon atoms, and Rc5 is hydrogen or an acid protecting radical; R ° 2a is hydrogen or an acid protecting radical such as methoxymethyl; R 0 (in one of several embodiments) may be a methyl group which may be substituted by a 5- or 6-membered aromatic heterocyclic ring having a single heteroatom such as 2- or 3-pyridyl, 2- or 3-thienyl or 2- or 3-furyl; and R 3 is a group of the formula R 4 SO 2 O- wherein R 4 may be alkyl, trihalomethyl or optionally substituted phenyl.

These compounds are said to be intermediates in the preparation of compounds in which the 3-substituent is a group of the formula 30 · 'R °

-CH = C-SR

which have been reported to have antibacterial activity.

il 84830

Although this patent excludes the possibility that R 0 is a methyl group having a nitrogen-containing heterocyclic ring as a substituent in both intermediates and end products, thus forming a heterocyclic-substituted propenyl group, it only mentions that the heterocyclic ring is attached to one heterocyclic ring. Thus, there is no reference to a propenyl group substituted with a quaternary ammonium group. The reference in the Example R ° intermediates and final products of Example 10 is only methyl. In addition, both the intermediate compounds and the final product must have a second substituent on the propenyl group (-03SR4 or -SR, respectively). Nor does it suggest or suggest that the 2-aminothiazol-4-yl substituent at position 15 be a 5-amino-1,2,4-thiadiazol-3-yl group.

European Patent Application 53,538 (published June 9, 1982) discloses e.g. 3-Vinyl-cephalosporin intermediates of formula (o) n 20 t N C — CONH-i-j H2N <^ V's' ^ N \ s ° Ν'νγί? Ί ^ CH = CH-R3 OR 1

COOR

Wherein n is 0 or 1, R 5 is hydrogen, alkyl, vinyl, cyanomethyl or an oxime protecting group, and R 3 is halogen. It does not suggest or suggest that the 2-aminothiazol-4-yl substituent be replaced by a 3-halo-1-propen-1-yl substituent.

The present patent application relates to a process for the preparation of novel cephalosporin derivatives which are effective antibacterial agents. More specifically, it relates to a process for the preparation of compounds of formula 12 8 4 830

n-1— c — comi -, - s'5 N

5 R1HN '^' s '' \ O CH = CH-CH®NsQ (I)

01.2 cocP

10 wherein R 1 is hydrogen or a common amino protecting group; R 2 is an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, propenyl or propargyl, or

-CH2-CH = CH2 or -CH2-C = CH

15

QsN-O is a quaternary ammonium group which is tri (lower alkyl) ammonio, N- (lower alkyl) pyrrolidinio, 2-methylthiazole or 2-amino-5-thiazolo [4,5-c] pyridinio or pyridinio. Which is optionally substituted (lower alkyl), amino, formylamino, carboxy, carboxy (lower alkyl) thio, hydroxy (lower alkyl), amino (lower alkyl), carbamoyl or N- ( lower alkyl) carbamoyl group, or an alkylene group having 3 to 5 carbon atoms attached at the 2,3-position, and their non-toxic pharmaceutically acceptable salts and physiologically hydrolyzable esters.

The process according to the invention is characterized in that a) a compound of formula (0) m N -r-C-CONH-t li "II] B2HN '^ \ s ^ no CH = CHCH:, Z (VI1) 35 Nor2 I! 2

υκ COOB

13 84830 wherein R 2 is as defined above, B1 is a common carboxyl protecting group, B2 is hydrogen or a common amino protecting group, Z is chlorine, bromine or iodine, and m is zero or one, is reacted with a tertiary amine Q = N (or successively with the secondary amine RR'NH and a compound of formula R "Z), and if m is 1, the sulfoxide is reduced by conventional means, followed by sequential removal of all protecting groups by conventional means; or b) a compound of formula 10

H2NtO

N \ j ^ n'CH = CHCH2 (^ N = Q (XI)

cocP

15 is reacted with an acid of formula 20

2 / rrcooH

B2HN N \ 2 25 or an acylating derivative of said acid, wherein R2 is as defined above, B1 is a common carboxyl protecting group and B2 is hydrogen or a common amino protecting group, and if desired, the compounds of formula I are converted into their non-toxic pharmaceutically acceptable salts 30 or its physiologically hydrolyzable esters.

The compounds to be prepared also include solvates (including hydrates) of the compounds of formula I as well as tautomeric forms of the compounds of formula I, e.g. the 2-imin-3-thiazolin-4-yl form of the 2-aminothiazol-4-yl group.

i4 84830

The invention also relates to certain intermediates used in the preparation of the compounds of formula I in said process.

As shown in the structural formula, the compounds of formula I may be in the "synH" or "Z" configuration with respect to the alkoxyimino group. Since these compounds are geometric isomers, little "anti-isomer" may be formed. The compounds of formula 1 prepared according to the invention contain at least 90% of the "syn" isomers Preferably, the compounds of formula I are formed as "syn" isomers, which are substantially free of the corresponding "anti" isomers.

In addition to possible geometric isomers with respect to the alkoxyimino group, compounds of formula I (and intermediate compounds of formulas VIII and IX) may also be formed, which are also geometric (cis and trans) isomers at the double bond of the propenyl group and such compounds and cis ("Z") that trans ("E") isomers are specifically included in the compounds to be prepared.

Non-toxic pharmaceutically acceptable salts of the compounds of formula I include salts with inorganic acids such as hydrogen chloride, hydrogen bromide, phosphoric acid and sulfuric acid, or organic carboxylic acids or sulfonic acids such as acetic acid, trifluoroacetic acid, citric acid, citric acid, citric acid with oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid, malic acid, p-toluenesulfonic acid and other known acids used in connection with penicillin and cephalosporins.

Examples of physiologically hydrolysable esters of the compounds of the formula I are indalyl, phthalidyl, methoxymethyl, acetoxymethyl, pivaloyloxymethyl, glycidyloxymethyl, phenylglycidyloxy 84830 methyl, 5-methyl-2-oxo-1,3 -dioxolen-4-ylmethyl esters and other physiologically hydrolyzable esters known and used in connection with penicillin and cephalosporins.

The compounds of formula I wherein R 1 is hydrogen have a very potent antibacterial activity against a variety of gram-positive and gram-negative bacteria and are useful in the treatment of bacterial infections in animals, including humans.

The compounds of formula I may be formulated for parenteral use in a conventional manner using known pharmaceutical carriers and excipients and may be presented in unit dosage form or in multi-dose containers. The compositions may take such forms as solutions, suspensions, or emulsions in oily or aqueous media, and may contain conventional dispersing, suspending or stabilizing agents. The compositions may also be in the form of a dry powder suitable for reconstitution prior to use, e.g. with sterile, pyrogen-free water. The compounds of formula I may also be formulated in the form of suppositories using conventional suppository masses such as cocoa butter or other glycerides. The compounds of the invention may, if desired, be administered in conjunction with other anti-biotics such as penicillins or other cephalosporins.

When presented in unit dosage form, the compositions preferably contain from about 50 to about 1,500 mg of the active ingredient of formula I. The dosage of the compounds of formula I will depend on factors such as the weight and age of the patient, as well as the specific nature and severity of the disease, and will be at the discretion of the physician. However, the dose used to treat an adult human is usually about 500 to 5,000 mg per day, depending on the frequency of administration and the route of administration. When administered intramuscularly or intravenously to an i6 84830 adult, a sufficient total dose is usually about 750 to 300 mg per day in divided doses, although higher doses of some compounds may be desirable in the case of Pseudomo-5 nas infections.

Preferred compounds of the invention are 1) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (trimethylammonio) -1 -propen-1-yl] -3-10 cephem-4-carboxylate, 2) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] —3- [3- (1-methylpyrrolidino) -1-propen-1-yl] -3-cephem-4-carboxylate, 3) 7- [2- (5-amino-1,2,4-thiadiazole -3-yl) -2-methoxyiminoacetamido] -3- [3-pyridinio-1-propen-1-yl] -3-cephem-4-carboxylate, 4) 7- [2- (5- Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-aminopyridinium) -1-propen-1-yl] -3-cephem-4- carboxylate, 5) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-formylaminopyridino) -1- propen-1-yl] -3-cephem-4-carboxylate, 6) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3 - [3- (3-aminomethylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 7) 7- [2- (5-amino-1,2,4-thiadiazol-3- yl) -2-meth oxyiminoacetamido] -3- [3- (3-carbamoylpyridino) -1-propen-1-yl] -3-cephem-4-carboxylate, 8) 7- [2- (5-amino-1,2,2 4-Thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridino) -1-propen-1-yl] -3-cephem-4-carboxylate, 9) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (2-methylthiazolio) -1-propen-1-yl ] - 3-cephem-4-carboxylate, 35-10) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyimino] 8 4 8 30 noacetamido] - 3- [3- (2-amino-5-thiazolo [4,5-c] pyridino) -1-propen-1-yl] -3-cephera-4-carboxylate, 11) 7- [2- (5- Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-hydroxymethylpyridinium) -1-propen-5-yl] -3-cephem-4 carboxylate, 12) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-hydroxymethylpyridinium) -1- propen-1-yl] -3-cephem-4-carboxylate, 13) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] - 3- [3- (4-N-methylcarbamoylpyride) nio) -1-propen-1-yl] -3-cephem-4-carboxylate, 14) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2- Methoxyiminoacetamido] -3- [3- (2,3-propylenepyridino) -1-propen-1-yl] -3-cephem-4-carboxylate, 15) 7- [2- (5-amino- 1,2,4-Thiadiazol-3-yl) -2-ethoxyiminoacetamido] -3- [3- (4-carbamoylpyridino) -1-propen-1-yl] -3-cephem-4- carboxylate, 16) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-cyclopentyloxyiminoacetamido] -3- [3- (4-carbamoylpyridinium) -1-20 propen-1-yl] -3-cephem-4-carboxylate, 17) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-propenyloxyiminoacetamido] -3 - [3- (4-carbamoylpyridinium) -1-propen-1-yl] -3-cephem-4-carboxylate, 18) 7- [2- (5-amino-1,2,4-thiadiazole-3 -yl) -2-propargyl-oxyiminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 19) 7- [2 - (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carboxypyridinium) -1-propen-1-yl] -3- cephem-4-carboxylate, 20-20) 7- [2- (5-amino-1,2,4-thiadiazole) 1-3-yl) -2-ethoxyiminoacetamido] -3- [3- (4-carboxypyridino) -2-propen-1-yl] -3-cephem-4-carboxylate, 21) 7- [2 - (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-carboxymethylpyridinium) -1-propen-3-yl] -3-yl] -3 -cephem-4-carboxylate and 84830 22) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4 -karboksimetyylitiopyridinio) -l-pro-pen-l-yl] -3-cephem-4-carboxylate.

The numbering system used here for the various reagents, intermediates and final products is as follows: [Roman - Arabic number Letter number] (if applicable) (if applicable) 10 "J * -

The Roman numeral denotes either the final product [I] or the intermediate compound or other reagent [all other Roman numerals]. Arabic numerals and letters 15 are not used in cases where the general class (species) of compounds is meant.

The Arabic numeral denotes a certain meaning of the substituent R2. If a particular R2 group has a carboxyl group protected by a common carboxyl protecting group, a comma (') is used after the Arabic numeral to indicate this. A "comma" is not used if the carboxyl group is unprotected. A "comma" is also used with a general R 2 substituent (i.e., R 2 ') to refer generally to an R 2 group having a protected carboxyl group.

25 The letter at the end of the compound number refers to a specific quaternary ammonium group: Φ

-NhQ

30 meaning.

For convenience, Arabic numerals and letters are shown below for a few primary R2 groups and quaternary ammonium groups.

is 84830

Arabic numeral_R * _ 1 = ethyl 2 = ethyl 3 = allyl 5 4 propargyl 5 = cyclopentyl

Kir_ ain_-N = Q_ A 1-methylpyrrolidino 10 B pyridine C = 2-amino-5-thiazolo [4,5-c] pyridine D = trimethylammonium E = 3-aminopyridine 15 F 3-formylaminopyridine G = 3-carbamoylpyridine H = 4-carbamoylpyridine I = 3-aminomethylpyridine J = 2-methylthiazole 20 K 3-hydroxymethylpyridine L = 4-hydroxymethylpyridine M 4- (N-methylcarbamoyl) pyridine N = 4-carboxypyridine 25 O 2,3-propylene pyridine carboxymethylpyridine Q 4-carboxymethylpyridine

In the primary evaluation of the compounds of formula I, the minimum inhibitory concentrations (MICs) of the compounds were determined by a two-fold agar dilution method on Mueller-Hinton agar against 32 species of test organisms in six groups. The geometric mean MICs determined in these experiments are shown in Table 1.

2o 84830

table 1

Compound Configuration Geometric MIC (pg / ml)

n: ° bond ° 1S_ (G +) ”Ia <G +) ~ Ib (G -) - Ia (G -) - Ib (G -) - II (G -) - III

for (5) (5) (5) (5) (5) (7) I-1A E / Z = 1/1 0.26 0.70 0.05 0.15 0.23 2.4 I-1A E / Z = 7/1 0.13 0.35 0.029 0.05 0.17 1.4 I-1B E 0.20 0.40 0.016 0.044 0.11 1.6 I-1B E / Z = 1 / 4 0.35 0.80 0.05 0.11 0.35 3.5 I-1C E 0.10 0.20 0.0071 0.033 0.087 3.8 I-1D E / Z = 1/1 0.61 1.4 0.10 0.26 0.46 2.4 I-1E E 0.20 0.40 0.0094 0.029 0.10 1.4 I-1F E 0.15 0.40 0.0094 0.033 0.099 1.2 I-1G E 0.20 0.35 0.094 0.033 0.10 1.4 I-1H E 0.20 0.40 0.013 0.043 0.10 0.97 I-1I E 0.80 1.6 0 .0 0.20 0.69 3.1 I-1J E 0.17 0.35 0.025 0.076 0.15 1.6 I-1K E 0.35 0.80 0.029 0.044 0.20 3.5 I-1L E 0.26 0.61 0.029 0.088 0.15 2.6 I-1M E 0.35 0.70 0.029 0.10 0.17 2.3 I-1N E / Z = 7/1 1.2 1 6 0.013 0.066 0.30 5.7 I-IO E 0.17 0.35 0.029 0.033 0.11 14 I-2H E 0.20 0.40 0.014 0.057 0.15 1.4 I-2N E 1.2 2.1 0.016 0.11 0.35 4.7 I-2N Z 1.4 3.1 0.044 0.15 0.69 10 I-3H E 0.23 0.40 0.057 0.10 0.52 1, 9 I-4H E 0.26 0.46 0.066 0.11 0.60 2.6 I-5H E 0.13 0.40 0.20 0.46 2.1 4.2 I-1P E 0.8 1.6 0.013 0.087 0.34 14 I-1Q E 0.7 0.92 0.0095 0.044 0.23 14 2i 84 8 30 (G +) - Ia: Penicillin-sensitive S. aureus (5 strains) (G +) - Ib: Penicillin-resistant S. aureus (5 strains) (G -) - Ia: Cephalothin-sensitive E. coli (2 strains), Kl. pneu monete (1 strain) and Pr. mirabilis (2 strains) 5 (G -) - Ib: Cephalothin-resistant E. coli (3 strains) and Kl.

pneumoniae (2 strains) (G -) - II: M. morganii (1 strain), Ent. clocae (2 strains) and Ser. marcescens (2 strains) (G -) - III: Ps. aeruginosa (7 strains) 10

The following Table 2 indicates the protective dose (PD50) in mice against selected microorganisms for a few compounds of formula I.

Table 3 shows the blood values of various compounds of formula I when administered intramuscularly to test mice at a dose of 20 mg / kg.

Table 2 _PD50_ S. aureus E. coli P. aeruginosa

Compound No._Smith_Juhl_A9843A_ I-1B 0.44 0.028 7.7

I-1B 0.65 0.072 NT

: 'V 25 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

30 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

- 35 NT = not tested 22 8 4 8 30

Table 3

c ~ .. T1 / 2 AUC

Compound No. 1 (pg / ml) (min) _ (pg t / ml) 5 I-1B 17 21 11 I-1C 21 32 18 I-1D 20 19 11 I-1H 23 16 14 I-1J 19 26 9 .7 10 I-1K 24 14 14 I-1M 20 23 14 I-1N 24 19 18 1-10 28 32 17 I-2N 22 20 12 15 I-3H 19 47 25 I-4H 27 22 16 I-5H 22 The preparation of the compounds of formula I is detailed in Reaction Schemes Ia, Ib and Ie and in Reaction Scheme 2 below.

Thus, the abbreviation "Ph" represents a phenyl group, thus the -CH (Ph) 2 moiety is a benzhydryl group, which is preferably a carboxyl protecting group, If R2 has a carboxyl group, it is desirable that the carboxyl group be protected with a conventional carboxyl protecting group such as t- Y represents chlorine, bromine or iodine.

23 84830

Reaction scheme la

H _N ^ S

rr ί 0i-NY ^ CH2Cl COOCH (Ph) 2

N -] - C-COOH

Il 11 II

H? N c ^ N N (111) \ N 2

OR

V

h il jpCONH — j "j H2N'J ^ S ^ N N \ CH2C10R COOCH (Ph) 2

Nai it KI N /

N - Γ— C-CONH - S 'S' 'N

: l | '{.Il> I P (Ph) -j S / N N ^ - N / A. 3

::: H2N \ O CH2I

f) D 2 ': COOCH (Ph) - (v); p (ph). / 3 ΙΔ v 24 84830 N -] - C-CONH -f S N. £) AS ^ N I j_N ^ ¢ 1 © IVI> H2N ° ^ j ^ S'CH2P (Ph) 3 OR2 COOCH (Ph) 2 base

'V

✓ S \ N-p- C-CONH —i-r ^ AS ^ -N \ (VII) H2N ö N ^ 0 y ^^ CH = P (Ph) 3 OR2 COOCH (Ph) 2 C1CH2CHO

V

N --- C-CONH —1-S 'S N

AS-N I 0J_hvJs (VIII> H2N N ^ 0 ^ CH = CHCH2C1 or2 COOCH (Ph) 2

- That's CI

V

25 S 4 o 3 O

S ^

N-- OCONH —-S

X '1 | M J αχ »H2N S 0 \ CH = CHCH2l OR2 COOCH (Ph) 2 R \ / \ HN \ R '\ secondary amine \

V

ii ^ r-rc0NHT ~ fSxi, XI1 I J_M p tertiary- h2n Νχ O ch = chch2-n ^ nen amilni

OR2 COOCH (Ph) R

(X)

R " '

V

r Deprotection of ~ n— rcoNH η — r ^ / H2 ^ -N i OR2 COOCH (Ph) 2 (XII)

V

N - j-C-CONH —-f Y

a.s-n i ® h2n N 0 oh = chch2-n = q \ 2 Θ

; OR COO

26 8 4 8 30

Reaction Scheme Ia shows two alternative means of obtaining Compound IX from Compound XII according to the invention.

The direct route using tertiary amine (XI) 5 is suitable for the preparation of all compounds of formula I.

In an indirect manner, a secondary amine is used as a reagent via compound X, and the quaternization takes place in the next step. The secondary amine RR'NH may be acyclic (e.g. dimethylamine) or cyclic (e.g. pyrrolidine), and this indirect process is therefore suitable for the preparation of compounds of formula I in which the quaternary ammonium group is an acyclic or "mixed" acyclic / cyclic combination. This indirect route is not suitable for the preparation of compounds of formula I in which the quaternary nitrogen is present in a completely unsaturated heterocyclic ring (e.g. pyridinio, thiazolio, 2-amino-5-thiazolo [4,5-c] pyridinio, and the like). ).

27 84830

Reaction Scheme lb (II)

^ ^ r- CHO

V

CH = N - ^> (XIII) QJ N \ ^^ CH2C1 \ COOCH (Ph) 2 \

Nai tai ΚΙ N.

V

CH;:; N-- QX- N \ <^ CH2I P (Ph) 3 <XIV) COOCH (Ph) 2 P (Ph) 3 (I / / ”^) - CH = N —- γθ ^ - - :, _n A © (xv) 0 ^ CH2P (Ph) 3 COOCH (Ph) 2 ....: base

.-: V

O- ™ Tj ‘X

A- N

: -. Ο CH = P (Ph) 3 COOCH (Ph) 2 28 84 830 Reaction Scheme le

ft -CH = N-j-S S N

\ - / I I (XVII) 0? N \> ^ CH = CH-CH2C1

COOCH (Ph) 2 Nai or KI

C ^ c ^ -pT5 '] Ύ CH = CH-CH2I COOCH (Ph) 2

/ R 1 HN

^ R

secondary amine V

\ __ / CH N j ^ I (XI), vvX O CH = CHCH9- <R tertiary- (XX) I 2 \ R | nen amine

.! R " '

<^> - ch = n— | - ^ /

: O ^ - CH = CHCH2? N = Q

(XXI) COOCH (Ph) 2

V

: T: 1 © (XXII> QS? - N \ ^^ CHC = CHCH2-N = Q:. ·. COO ©:: N-acylation

with compound III

V

29 84830

η ~ Π | C "C0NH Ί ΓS

«2“ S '\ N'ViJNCH = CHCH2-NHQ

OR2 coo0 Reaction Scheme Ic is a further variation of Reaction Scheme 1b.

In Reaction Schemes 1a and 1b, the quaternization of the 3-side chain is the last step, but in Reaction Scheme le the last step is the acylation of the 7-amino group.

The relationship between reaction schemes 1a, Ib and 1e is shown in the following flow chart.

30 84830 y UI) COOCH (Ph) 2 \ / \! B "f \ i N - jT-C-CONH —- SS C- CH NJ ζ h2A ^ \ 2JXAch2C1 (xiii) 0 \ cn ^ 2 OR2 I COOCH (Ph) .p (IV) COOCH (Ph) 2 \ / - lb la

::. H2NtO

v 7-N acylation ^ Ο CH = CHCH „C1 S <------ I 2 N-jp-C-CONH-1- ^ N (XVIII) COOCH (Ph) 2 HN ^ S 'Nv ° ^ ~ NV ^ OH = CHCH ~ Cl 2 \ or2 T 2 (VIII) COOCH (Ph) 2: -. Quaternization Quaternization: .-. la Protection Removal of protection lb removal: .-. ν '

Ψ S

H2N — Γ ~ ί ^ ;;;; Compound z ^ - .. yiomti </ - YkCH = CD ® lc coo®: (XXII) 3i 84830

In Reaction Schemes Ia, 1b and 1e, the benzhydryl group is shown as the primary carboxyl protecting group. It will be readily apparent to those skilled in the art that other carboxyl protecting groups well known in the art may be used.

Acylating acid III can be used in the form of a derivative such as its acid halide, active ester, mixed acid anhydride, etc., all of which are well known in the art. Its use as its hap-10 pokloridin is recommended. Also, the amino group III of the acylating acid may be protected with any common amino protecting groups, e.g., N-trityl, N-formyl, or the like.

The base used to convert the phosphonium iodide (VI or XV) to the phosphorylide (VII or XVI) may be NaOH,

Na 2 CO 3, TRA-410 (OH ") resin, IRA (CO 3 =) resin or a similar base or a mixture thereof.

The chloroacetaldehyde used to convert phosphorylide VII to 3-chloropropenyl-3-cephem compound VIII (or to convert compound XVI to compound XVII) may be a commercially available 40-50% aqueous solution, distilled solution (e.g. 70%) or anhydrous aldehyde.

It was found that the typical Z: E ratio of Compound VIII prepared from Compound VII (Scheme Ia) at the propylene-two-bond bond was about 2: 1. On the other hand, compound VIII prepared from compound XVIII (Scheme Ib) was typically almost exclusively the z-isomer. The difference cannot be due to the route used, but to the conditions used in the Wittig reaction (VII -> VIII or XVI -> XVII). It was also found that the use of a suitable silyl reagent such as N, O-bis- (trimethylsilyl) acetamide in the Wittig reaction (VII -> VIII in Scheme Ia and XVI -> XVII in Scheme Ib) resulted in improved yields and purity of compounds VIII and XVIII. . The reaction is preferably carried out using 2-5 equivalents of silyl reagent.

32 8 4 8 30

When the chloropropenyl cephem compound (VIII) was reacted with NaI in acetone to give the iodopropenyl cephem compound (IX), the double bond in the propenyl group was isomerized from Z to E during iodination. With a short reaction time, the configuration of the parent compound VIII was retained in large amounts, with the long reaction time primarily producing the E-isomer of compound IX. If the reaction time is too long at high temperature, compound IX is obtained, but less pure. It was found that in about 10 to 10 minutes at 25 ° C and in 2 hours at 5 ° C, pure compound IX is obtained in good yields. Using Reaction Scheme le, it was found that iodination of XIV with NaI yields a purer compound if the acetone solution is diluted with CCl 4 with the iodination substantially complete and the isomerized portion of the reaction mixture is passed into acetone-CCl 4.

When iodination of the chloropropenyl cephem compound (XVII) to the iodopropenyl cephem compound (XIX) was performed with KI in DMF, the isomerization of the double bond from Z to E ta-20 occurred as rapidly as the iodination. The whole reaction was completed in 45 minutes at room temperature to give pure compound XIX without dilution during the reaction with CCl 4.

Compound XII was deprotected normally without purification and the final product was purified by reverse phase column chromatography using a glass column packed with Waters' Associates PreoPAK-500 / C18 cartridge.

‘* 33 84 830

Reaction Scheme 2 «—Π-C“ CONH -j- (XXIII)

R HN 0 XcH = CH-CH2I

OR2 COOCH (Ph) 2 0 N —ίΓ ~ C-C0NH -yX s "Ί (XXIV) R1HN '^ V * S' 'N Νν'γ ·' '' ^ ΟΗ <: Η-ΟΗ2Ι ^ 0R2 COOCH (Ph ) 2 V Ϊ N -! - C-CONH —I- (S ^ n ll I 1 <=, <xxv) R1HN / ^ 's''NN NY ^ ch = ch-ch2n = q

Nvor2 COOCH (Ph) 2: · reduction Ψ N-1-C-CONH —i- {> 3® || Il k I I φ <XXVI>

r1hnA-S-N \ 0 <* - N'v ^ XcH-CH-CHjNSQ

'or2 COOCHlPh) 2 deprotection ... ψ

:. · N - j-C-CONH —-S S 'N

ls, «t ® (I) h2n ö n o vv ^ ^ ch = ch2nsq> 9 ö OR COtT '34 8 4 8 30

Reaction Scheme 2, shown above in the form of a brief outline, is otherwise similar to Reaction Scheme Ia, except that Compound XXIII (corresponding to Compound IX in Reaction Scheme Ia) is converted to its S-oxide prior to quaternization. Compound XXV is then reduced, and the remainder of Reaction Scheme 2 is similar to Reaction Scheme Ia. Reaction Scheme 2 primarily recommends protecting the 7-side chain amino group with a known amino protecting group such as a trityl group.

The acylating acids of the formula III used in this case are either known compounds or can be easily prepared by known methods.

European Patent Publication 7,470, published October 12, 1980 (patent application published February 6, 1980), exemplifies the preparation of compounds of formula III wherein R 2 is methyl, ethyl, propyl and isopropyl.

U.S. Patent No. 4,390,534, cited above, in the prior art description mentions by way of example the preparation of a wide variety of compounds of formula III wherein R 2 is, for example, cyclopentyl, 2-cyclopenten-1-yl, allyl, 2-propynyl, 1-tert-butyloxycarbonyl -1-methylethyl, 1-tert-butyloxycarbonyl-1-cyclopentyl, 1-ethoxycarbonyl-1-methylethyl, tert-butyloxycarbonylmethyl, 1-tert-butyloxycarbonyl-2-methylpropyl, trityl and their similar group.

The compound II (7-amino-3-chloromethyl-3-cephem-4-carboxylate) mentioned herein, which was used as a starting material in Reaction Schemes Ia, Ib and Ic, is a known compound.

The tertiary amines of formula XI (and the secondary amines RR'NH) used in the preparation of the quaternary ammonio compounds of formula I are either known compounds or are readily prepared by those skilled in the art. Many amines are available in 35 commercial products.

35 84830

The reactions of the process of the invention are carried out in an anhydrous organic solvent such as dimethyl sulfoxide, hexamethylphosphoramide, methylene chloride, chloroform, ethyl ether, hexane, ethyl acetate, tetrahydrofuran, acetonitrile and the like, and the like. The reactions are conveniently performed at a temperature of about -10 to + 50 ° C; it is usually recommended to carry out the reactions at room temperature. During the quaternation step, at least 1 mole per mole of compound IX, XIX, XXIII or XXIV must be used; it is usually recommended to use about 25 to 100% excess tertiary amine.

Carboxyl protecting groups suitable for use as the R1 group in the above reactions are well known to those skilled in the art and include aralkyl groups such as benzyl, p-methoxybenzyl, p-nitrobenzyl and diphenylmethyl (benzhydryl); alkyl groups such as t-butyl; haloalkyl groups such as 2,2,2-20 trichloroethyl and other carboxyl protecting groups described in the literature, e.g. GB 1 399 086. Priority is given to the use of carboxyl protecting groups which are readily removed by acid treatment. Particularly preferred carboxyl-protecting groups are benzhydryl and t-butyl groups.

. Amino protecting groups suitable for use as a B2 group are also well known in the art and include a trityl group and acyl groups such as chloroacetyl, formyl and trichloroethoxycarbonyl. Preference is given to amino protecting groups which can be easily removed by treatment with an acid, e.g. a trityl group.

When the cephalosporin core is used in the form of 1-oxide (m = 1), 1-oxide is prepared by known methods such as oxidation with m-chloroperbenzoic acid, per-35 acetic acid, sodium tungstate, etc. The 1-oxide can then be reduced by known methods, e.g. an alkoxysulfonium salt with an iodide ion in an aqueous medium. The alkoxysulfonium salt itself is readily prepared by treating the 1-oxide with, for example, acetyl chloride.

The invention also relates to novel intermediate compounds of the formula N-n-C -CONH-i- 10 Π N 11 j_N i

S-X N \ CH = CH-CH-Z

OR I 1

cooB

wherein R 1 is hydrogen or a common amino protecting group, R 2 is an alkyl group having 1 to 4 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, propenyl or propargyl, B 1 is hydrogen or a common carboxyl protecting group and Z is chlorine, bromine or iodine; or a salt, hydrate, solvate or ester thereof.

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, etc.); aroyl (e.g. benzoyl, etc.), (lower) alkanesulfonyl (e.g., mesyl, ethanesulfonyl, etc.) or arylsulfonyl (e.g., benzenesulfonyl, tosyl, etc.) As used herein, the terms "(lower) alkyl", "(lower) alkoxy" , "(lower alkylthio" (or similar terms) means straight or side chain alkyl, alkoxy, alkylthio groups (or the like) having 1 to 6 carbon atoms, inclusive. Similarly, the terms "(lower) alkenyl "and" (lower) alkynyl "mean alkenyl or alkynyl groups having 35 to 6 carbon atoms.

37 84830

Example 1 njc-conh —-r '"S ^ i _N * 0 / ^ 5 h2n ^ s" \ jX \ Ch = ch-ch2-n ^^ OCH3 COOe CH3 1-1A * Z / E = 1/1 10 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (1-methylpyrrolidino) -1-propen-1-yl] - 3-Cephem-4-carboxylate (I-1A) To a solution of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] - 3-15 (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (Z / E = 2/1) (150 mg, 0.21 mmol) in ethyl acetate (2 ml), a solution of 1-methylpyrrolidone (36 mg, 0.042 mol) in ethyl acetate (1 ml) was added 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 solid (130 mg), formic acid (1 mL) and concentrated HCl (0.1 mL) was stirred at room temperature. After 1 h, the reaction mixture was concentrated in vacuo, diluted with water (20 mL) and filtered. The aqueous solution was passed through a reversed phase column (PrepPAK-500 / C18 cartridge pack, 100 mL) eluting with water and 10% CH 3 OH. The desired fractions were collected and concentrated in vacuo to a small volume and lyophilized to give 13 mg (12% of the title compound (I-1A) (Z / E = 1/1), melting at> 280 ° C (dec).

IR: KBr cm-1 \ make 3400, 1760, 1660 and 1610.

UV: λmax (phosphate buffer, pH 7) nm (E1 * 1 cm) 236 (372),

288 (322). _1_H

35 NMR: D 2 O δ ppm: 2.31 (4H, m, N), 3.12 (3H, s, 38 84 3 30 N-CH 3), 3.6 (5H, m, 2-H & |), 3.79 (1H, s, 2-H), 4.1 (2H, d, J = 8, CH 2 N), 4.2 (3hT 2 s, OCH 3), 5.36 (1H, d, J = 4 , 5, 6-H), 5.95 (3H, m. 7-H & 3-CH = CH), 6.66 (1 / 2H, d, J = 5 10, 3-CH cis) and 7, 10 (1 / 2H, d, J = 16, 3-CH trans).

Example 2 N-j-C-CONH —J — r η J | N II J1 A * 10 i, 2n ^ s "N 0 ch-ch-ch2-n ^ 0CH3 COOe

I-1B * E

7- [2- (5-amino-1,2,4-thlazolol-3-yl) -2-methoxy-iminoacetamido] -3- [3-pyridinio-1-propen-1-yl] -3-cephem -4-carboxylate (I-1B)

Mixture of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl ) -3-cephem-4-carboxylate (IX-1) 20 (E, 716 mg, 1 mmol), pyridine (158 mg, 2 mmol) in dimethyl sulfoxide (DMSO) (1 mL), was stirred for 1 h at room temperature. Ethyl acetate (20 ml) was added to the mixture to precipitate a solid (620 mg), which was added to formic acid (6 ml) with sodium hydrogen sulfite (60 mg). The mixture was stirred for 30 minutes at 40 ° C and evaporated to dryness. The residue was dissolved in water (40 ml) and some insoluble matter remained. The aqueous solution was applied to a reverse phase column (PrepPAK-500 / Cie, 100 mL), eluted with water (300 mL) and 5% aqueous CH 3 OH (800 mL), and the eluate was monitored by UV (254 nm) and By HPLC. Fractions (5% aqueous CH 3 OH solution) containing the desired product were combined, evaporated to a small volume and lyophilized to give 40 mg (8%) of the title compound (I-1B), m.p.> 200 ° In C (decomposing).

39 84 330 IR: KBr cm-1 Vmake 3350, 1760, 1660 and 1600.

UV: ^ make. (phosphate buffer, pH 7) run (E1 \ cm) 240 (352), 258 (366), 267 (279) and 290 (469).

NMR: D 2 O + DMSO-d 6 δ ppm 3.74 (2H, br - s, 2-H), 4.20 (3H, 5 s, OCH 3), 5.92 (1H, d, J = 4.5, 7-H), 6.15 (1H, m, 3-CH = CH), 7.094 (1H, d, J = 16, 3-CH trans), 8.2 (2H, m, Py-H4) and 8 .97 (2H, m, Py-H26).

Example 3 10 N-η-C-CONH -] - f S ^ h2 (As'N o-LnY ^ ch.ch-ch2® ^> 0CH3 COO® I-IB * Z / E = 4/1 15 7- [2- (5-amino-l, 2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3-pyridinio-l-propen-2-yl] -3-cephem-4- carboxylate (I-1B)

Chloropropenyl compound, diphenylmethyl-7- [2- (5-20 amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-chloro-1-propen-1-yl) - 3-Cephem-4-carboxylate (VIII-1) (Z, 937 mg, 1.5 mmol) was added to a stirred solution of pyridine (237 mg, 3 mmol) in DMSO (3 mL) with Nai (11 mL). mg, 0.075 mmol). The mixture was allowed to stand overnight at room temperature in the dark.

The mixture was diluted with ethyl acetate (10 mL) and dried to give 350 mg of the protected product. The precipitate was treated with formic acid (3.4 ml) with sodium bisulfite (34 mg) for 30 minutes at 40 ° C. After removal of formic acid, the residue was purified by reverse phase column chromatography (packing PrepPAK-500 / Cie cartridge, 100 mL) eluting with 5% aqueous CH 3 OH. The fractions containing the desired product were combined by HPLC analysis, evaporated to dryness in vacuo and freeze-dried to give 41 mg (5.5%) of the title compound (I-1B) (Z / E = 4/1). Sp. > 200 ° C (decomposed).

IR: KBr cm-1 N max 3300, 1760, 1660 and 1600.

UV; Amax (phosphate buffer, pH 7) nm (E1 * a cm) 237 (386), 5,250 (377), 258 (369), 265 (347) and 280 (311).

NMR: D 2 O δ ppm 3.45 & 3.76 (each 1H, d, J = 16, 2-H), 4.18 (3H, d, OCH 3), 5.34 (3H, m, CH = CH- CH 2 & 6-H), 5.92 (1H, d, J = 4.5, 7-H), 6.58 (4 / 4H, d, J = 11, 3-CH cis), 7.03 ( 1 / 5H, d, J = 16, 3-CH trans), 8.12 (2H, m, Py-H35), 8.56 (1H, m, Py-H4) and 8.82 (2H, m , Py-H26).

Example 4 .S.

N-p- C-CONH -j-p ^ 15 Η2Η ^ 3 · 'Ν t 0J — l) vyACH, CH-CH2V ^ j— »XOCH3 coo0 k ^ X * S’S> NlH2

I-1C * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (2-amino-5-thiazolo [4,5-c] [pyridino] -1-propen-1-yl] -3-cephem-4-carboxylate? ti (I-1C): A stirred solution of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (E-isomer, 714 mg, 1 mmol), 2-aminothiazolo [4,5- c] pyridine [prepared according to the method of T. Hakahashi et al .; Pharm. Bull (Japan) 2 (1954), p. 30 34] and dry DMSO (1 mL) were kept at room temperature for 1 hour. Ethyl acetate (20 ml) was added to the reaction mixture to give a yellow powder (710 mg). To the powder (700 mg) were added formic acid (7 ml) and sodium hydrogen sulfite (70 mg), and the mixture was stirred for 30 to 35 minutes at 40 to 45 ° C. After evaporation to dryness, 4i 84 8 30 of the residue were triturated with water (40 ml). The insoluble matter was filtered off and the filtrate was chromatographed on a reverse phase column (PrepPAK-500 / C10, 100 ml) with water and 10% CH 3 OH as eluent. The fractions containing the desired product 5 were combined and the solvent was removed in vacuo. Freeze-drying gave the desired product (Ι-Ιο) as a colorless amorphous E-isomer powder. Yield 110 mg (19%). Sp. > 200 ° C (dec.

IR: KBr cm'1 V> max 3300, 1760, 1660, 1630 and 1600.

10 UV: Anaks. (phosphate buffer, pH 7) nm (E1, 11cm) 245 (499) and 285 (286).

NMR: DMSO-d 6 + D 2 O δ ppm 3.86 (3H, s, OCH 3), 4.98 (1H, d, J = 4.5, 6-H), 5.2 (2H, m, CH = CH -CH 2), 5.57 (1H, m, 3-CH = CH), 5.96 (1H, m, 7-H), 7.16 (1H, d, J - 16, 3-CH trans), 8.36 15 & 8.45 (1H, d, J = 7, Py-H each) and 8.92 (1H, s,

Pooh).

Example 5

N -C-CONH-1-S S

20 ^ * ® H „NN 0 ^ ^ CH = CH-CH_-N (CH,) _ noch I θ 33 3 cooö I-1D * Z / E - 1/1 ·: · 25 7— [2 - (5 -amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-trimethylammonio-1-propen-1-yl) -3-cephem-4-carboxylate (I-1D )

To a solution of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-30 (3-iodo-1-propen-1-yl yl) -3-cephem-4-carboxylate (IX-1) (Z / E = 2/1; 490 mg, 0.68 mmol) in ethyl acetate (14 mL) was added 0.1 M ethereal trimethylamine (13.6 mL). ) in a single installment. The mixture was stirred for 10 minutes and evaporated to dryness and the residue triturated with ether 35 (20 ml). The resulting solid (490 mg) was added to trifluoroacetic acid (0.2 mL) with one drop of anisole. After stirring for 1.5 hours, the mixture was evaporated to dryness in vacuo and the residual oil was triturated with ether (20 ml). The resulting precipitate was collected by filtration and dissolved in water (20 ml). Minor solids were removed and the aqueous solution was eluted on a C18 reverse phase column (packed PrepPAK-500 / C18 cartridge; Waters; 30 ml) using water as eluent. Fractions containing the desired compound were combined and concentrated to a small volume and lyophilized to give 30 mg (9.2%) of the title compound (I-1D) (Z / E = 1/1) as a colorless amorphous powder, melting> 150 °. In C (decomposing).

IR: KBr cm-1 Vmax 3300, 1770, 1670 and 1605.

15 UV: Aaek ·. (phosphate buffer, pH 7) nm (E1,; lcm) 236 (389) and 287 (343).

NMR: D, δ ppm 3.45 & 3.7 (1H, d, J = 16, 2-H), 3.81 (1H, δ s, 2-H), 4.1 (2H, d, J = 8, -CH 2 N), 4.21 (3H, s, OCH 3), 5.39 (1H, d, J = 4.5, 6-H), 5.95 (2H, m, 3-CH = CH & 7-H), 6.61 (1 / 2H, d, J = 11, 3-CH cis) and 7.05 (1 / 2H, d, J * 16, 3-CH trans).

Example 6 N-n-C-CONH -j-S S ^ v - Ajl I '

0CH3 COO® W

I-1E * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-aminopyridinium) -1-propen-1-yl] ] -3-cephem-4-carboxylate (I-1E)

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-Cephem-4-carboxylate (IX-1) (E; 716 mg, 1 mmol) 43,84830 was added to a stirred solution of 3-aminopyridine (188 mg, 2 mmol) in DMSO (1 mL). The mixture was stirred for 1 hour and diluted with ethyl acetate (20 mL). The precipitate formed was collected by filtration, washed with ethyl acetate and dried to give 520 mg of a yellow powder. A mixture of powder (500 mg), formic acid (5 ml) and sodium hydrogen sulphite (50 mg) was stirred for 30 minutes at 40 ° C. The mixture was evaporated to dryness in vacuo, dissolved in water (40 ml) and filtered to remove insoluble material. The aqueous solution was chromatographed on a reverse phase column (packing PrepPAK-500 / C18, 100 mL) eluting with 7.5% aqueous CH 3 OH. Fractions containing the desired compound were evaporated to dryness and lyophilized to give the title compound (I-1E) (7 mg, 1.4%), melting at> 185 ° C (dec).

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

UV; ÄmakB. (phosphate buffer, pH 7) nm (E1, slcm) 246 (403) and 290 (468).

NMR: D 2 O δ ppm 3.72 (2H, m, 2-H), 4.14 (3H, s, OCH 3), 5.35 (3H, m, 6-H & CH = CH-CH 2), δ .9 (1H, d, J = 4.5, 7-H), 6.1 (1H, m, 3-CH = CH), 7.05 (1H, d, J = 16, 3-CH trans) , 8.1 (1H, m, Py-H5), 8.54 (1H, br-s, Py-H6), 8.68 (1H, m, y Py-H4) and 9.4 (1H, m , Py-H2).

Treatment of IX-1 (716 mg, 1 mmol) with 3-t-butoxycarbonylaminopyridine (324 mg, 1 mmol) in a similar manner to that described above afforded 12 mg (2.3%) of I-1E.

Example 7 30 N ΓΪ-C-CONH —I Γ ^ ΝΗ, i '1 li J_1 i * ®> r <

H, N ^ Nv O ^ X-Y ^ CH-CH-CHj-N J

: T: «» s ClxP

35 I-1E * X / E = 1/1 44 84 830 7- [2- (5-amino-1,24-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3- Amino-1-pyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate (I-1E) A mixture of diphenylmethyl 7- [2- (5-amino-5 1,2,4 -thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (Z / E = 3 / 2, 500 mg, 0.7 mmol) and 3-aminopyridine (66 mg, 0.7 mmol) in dimethyl sulfoxide (1 mL) were 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 in formic acid (3 mL) with concentrated HCl (0.3 mL) and stirred for 1.5 h at room temperature. The mixture was evaporated to dryness in vacuo. The residue was dissolved in 2% HCl (10 mL) and filtered. The aqueous layer was chromatographed on a reverse phase column (PrepPAK-500 / C18, 100 mL). After washing with water (500 mL), the column was eluted with 5% aqueous CH 3 OH. The fractions containing the title compound were combined, evaporated to dryness in vacuo and lyophilized to give 15 mg (4.2%) of the title compound (I-1E) (Z / E = 1/1) as a colorless amorphous powder. Sp. > 160 ° C (decomposed).

IR: KBr cm -1 vacancies 3400, 1765, 1675, 1620 and 1600.

UV: λmax (phosphate buffer, pH 7) nm (E1 cro) 244 (434) and 287 (333).

NMR: DMSO-d 6 + D 2 O δ ppm 3.73 (2H, m), 4.14 (3H, s, OCH 3), 5.35 (3H, m, 6-H & CH = CH-CH 2), δ, O (2H, m, 7-H & 3-CH = CH), 6.6 (1 / 2H, d, J = 11, 3-CH cis), 7.05 (1 / 2H, d, J = 16) , 3-CH trans), 8.08 (1H, m, Py-H5), 8.6 (2H, m, Py-H46) and 9.4 (1H, m, Py-H2).

45 84830

Example 8

N τι-C-CONH-i (^ NHCHO

Ji 'i I __' A * »/ r \ 5 h, n- ^ s" k 0 ^ —N \ ^ CH-CH-CH2-N ^ och3 cöo®

I-1F * E

7- C2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-10-iminoacetamido] -3- [3- (3-formylaminopyridinio) propen-1-yl] -3- Cephem-4-carboxylate (I-1F) A mixture of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3 iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) 15 (E, 716 mg, 1 mmol) and 3-formylaminopyridine [prepared according to the method of N. Enomoto et al. in accordance with; Bull. Chem. Soc. Japan, 45 (1972), p. 2665] (244 mg, 2 mmol) in DMSO (2 mL), was stirred at room temperature for 1 h, and poured into ethyl acetate (20 mL). The precipitate was collected by filtration, washed well with ethyl acetate and dried. A mixture of quaternized salt (500 mg) and sodium bisulfite (50 mg) in HClOH (5 ml) was stirred at 40-50 ° C for 80 minutes and evaporated to dryness in vacuo. The residue was dissolved in water (40 mL), neutralized with NaHCO 3 and then filtered to remove insoluble material. The clear filtrate was chromatographed on a reverse phase column (PrepPAK-500 / C18, 100 mL) with water and 5% CH 3 OH, 10% CH 3 OH, 20% CH 3 OH and 30% CH 3 OH. with CH 3 OH. Fractions containing the desired compound were combined, evaporated to dryness in vacuo and lyophilized to give 16 mg (2.9%) of the title compound (I-1F) (E) as a tan powder. Sp. > 170 ° C (decomposed).

IR: KBr cm -1 max 3340 (br), 1760, 1670, 1620 (br) and 1590.

46 8 4 8 30 UV: A. „akB. (phosphate buffer, pH 7) nm (EI), 218 (428), 248 (362) and 290 (474).

NMR: D 2 O + NaHCO 3 δ ppm 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH 3), 5.91 (1H, d, J = 4.5, 7-H), 6.25 (1H, m, CH = CH-CH 2), δ 6.98 (1H, d, J = 16, 3-CH trans), 8.8-7.9 (4H, m, Py-H) and 9.38 (1H, br, NHCHO).

Example 9 N "Ti, C" CONH T "f 8 Ί -rCONH 2 10 il N 1_N, ^ 1 * ^ '0 ^ -n \ ^ NCi..ch-ch2-n 0CH3 coo®

I-1G * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-carbamoylpyridinium) -1-propen-1-yl] -3-Cephem-4-carboxylate (I-1G) To a solution of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3 -20 (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (E; 716 mg, 1 mmol) in DMSO (2 mL), nicotinamide (244 mg , 2 mmol) and the mixture was stirred at ambient temperature for 1.5 hours and poured into ethyl acetate (200 ml) with stirring. The precipitate formed was collected by filtration. The quaternized salt (500 mg) was dissolved in HClOH (5 ml) in the presence of sodium bisulfite (50 mg) and the mixture was heated at 40-50 ° C with stirring for 40 minutes and evaporated to dryness. The residue was dissolved in water (40 ml) and the insoluble material was filtered off. The filtrate and washings were combined and chromatographed on a reverse phase column (PrepPAK-500 / C18, 100 mL) after sequential elution with water and 5%, 10 and 20% aqueous CH 3 OH. solutions, the fractions containing the desired substance were combined, evaporated to dryness in vacuo and lyophilized to give 21 mg (3.8%) of the title compound (I-1G) (E) as a yellow powder, mp> 175 ° C (decomposed).

IR: KBr cm-1 3340 (br), 1760, 1670 and 1600.

UV: λmax (phosphate buffer, pH 7) nm (E1C01) 235 (326), 5274 (sh, 405) and 290 (446).

NMR: D 2 O + NaHCO 3 δ ppm 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH 3), 5.32 (1H, d, J = 4.5, 6-H), 5.45 (1H, d, J = 7, CH = CH-CH 2), 5.88 (1H, d, J = 4.5, 7-H), 6.15 (1H, d - t, J = 16 & 7, 3-CH = CH), 7.00 (1H, d, J = 16, 3-CH trans), 8.23 (1H, m, Py-H5), 9.03 (2H, m , Py-H4a6) and 9.34 (1H, s,

Py-H2).

Example 10 N -j-C-CONH -T-f S 15 15 „: JL *„ N '^' sx n n \ ^^ ch = ch-ch2-n Vconh2 2 N ° CH3 COO®

I-1H * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridinium) -1-propen-1-yl] -3-Cephem-4-carboxylate (I-1H) To a stirred solution of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyimino-25] acetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (E; 716 mg, 1 mmol) in dry DMSO (2 mL) , isonicotinamide (244 mg, 2 mmol) was added. The mixture was stirred at room temperature for 1 hour and then poured into ethyl acetate (200 ml). The resulting precipitate 30 was collected by filtration, washed well with ethyl acetate and dried. A stirred mixture of quaternized material (400 mg) and sodium bisulfite (40 mg) in HClOH (4 mL) was heated at 40-50 ° C for 1 h and evaporated to dryness in vacuo. The crude solid 35 was dissolved in water (40 mL). After the insoluble material was filtered off, the filtrate was chromatographed on a reverse phase column (packing PrepPAK / C18, 100 ml) using water and 5-, 10-, 20- and 30% CH3OH in water as eluent. solutions of. Fractions containing the desired compound were combined, evaporated to dryness and lyophilized to give 21 mg (3.8%) of the title compound (I-1H) (E) as a pale yellow powder. Sp. > 180 ° C (decomposed).

IR: KBr cm-1 3340 (br), 1760, 1670 and 1600.

UV: ^ maka. (phosphate buffer, pH 7) nm (E1% 1 ca) 222 (362) and 10,285 (452).

NMR: D 2 O + NaHCO 3 δ ppm 3.68 (2H, br, 2-H), 4.15 (3H, s, OCH 3), 5.33 (1H, d, J = 4.5, 6-H), 5.46 (2H, d, J = 7, CH * CH-CH2), 5.90 (1H, d, J = 4.5, 7-H), 6.17 (1H, d - t, J * 16 & 7, 3-CH = CH), 7.02 (1H, d, J = 16, 3-CH trans) and 8.43 & 9.09 (each 2H, d, 3 = 7, Py-H ).

Example 11 N-j-j-C-CONII - ^ ^ 9H2NH2

H2N ^ S "N L

20 OCHj \ == /

I-1I * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-aminomethylpyridinium) -1-25 propen-1-yl] -3-Cephem-4-carboxylate (I-1I)

A mixture of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) ) -3-cephem-4-carboxylate (IX-1) (E, 716 mg, 1 mmol) and 3- (t-butyloxycarbonylamino-30-style) pyridine (526 mg, 2 mmol) in DMSO (2 mL) were stirred at ambient temperature. at 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 quaternized salt (500 mg) and sodium bisulfite (50 mg) in HCOOH (5 ml) was stirred at 49-48 ° C for 40 minutes and evaporated to dryness in vacuo. The remaining solid was dissolved in water (40 mL) and the mixture was neutralized with NaHCO 3. The insoluble matter was filtered off and the filtrate was chromatographed on a reverse phase 5 column (packed PrepPAK-500 / Cie cartridge, 100 ml) eluting successively with water and 5-, 10-, 20- and 30% aqueous CH 3 OH solutions. Fractions containing the desired compound were combined, evaporated to dryness and freeze-dried to give 10 mg (1.8%) of the title compound 10 (I-1I) (E) as a tan powder.

IR: KBr cm-1 V> maka 3380 (br), 1760, 1650 (sh) and 1620 (sh). UV: Amaka (phosphate buffer, pH 7) nm (E1 em) 235 (sh, 260) and 286 (370).

NMR: D 2 O + NaHCO 3 δ ppm 3.68 (2H, b 2, 2-H), 4.16 (3H, s, 15 CH 3), 6.98 (1H, d, J = 16, 3-CH trans), 8.05 (1H, m,

Py-H5), 8.50 (1H, m, Py-H4) and 8.80 (2H, m, Py-H26). Example 12 N-n-C-CONiI -T-S S ^ 1 20 11N ^ S ^ N n o ^ «Y <ch: ch-ch ^ coNh2 0CH3 C00®

Ι-1Η * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] -3- [3- (4-carbamoylpyridinium) -1-propen-1-yl] -3-Cephem-4-carboxylate (I-1H) Mixture of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3 - (3-30 iodo-1-propen-1-yl) -3-cephem-4-carboxylate (IX-1) (E-isomer; 4.1 g, 5.7 mmol) and isonicotinamide (1.4 g) , 11 mmol) in dry DMSO (6 mL) was stirred for 2 h at room temperature as monitored by TLC on a silica gel plate (CHCl 3 CH 3 OH = 3: 1). The reaction mixture was diluted with ethyl acetate (100 mL) to separate the yellow resin 84,830, which was treated with formic acid (40 mL) and sodium bisulfite (390 mg) for 30 minutes at 45 ° C. The resulting solution was evaporated to dryness. The residue was dissolved in water (100 ml), and the insoluble matter was removed by filtration. The combined filtrate and washings were applied to the top of the column with reverse phase packing (PrepPAK-500 / C18, 1120 mL). The column was eluted with water. 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 10, pH 7.2 with 20% CH30H). Fractions 4 and 5 were combined and evaporated to a small volume. Freeze-drying afforded 250 mg (8.1%) of the title compound (I-1H), melting at> 180 ° C (dec).

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

Preparation of hydrochloride

To a suspension of I-1H (98 mg, 0.18 mmol) in CH 3 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 form a precipitate which was collected by filtration, washed with acetone (2 x 10 mL) and dried in vacuo to give the hydrochloride salt of Compound I-1H as a colorless powder. Yield 88 mg (79%). Sp. > 190 ° C (dec.). IR: KBr cm'1 V> max 3300, 1770, 1680 and 1620.

UV: <Amake. (phosphate buffer, pH 7) nm (E1 *! ^) 227 (385) and 286 (374).

NMR: D 2 O δ ppm 2.32 (1H, s, acetone-H), 3.79 (2H, br - s, 2-H), 4.17 (3H, s, OCH 3), 5.34 (1H, d, J = 4.5, 6-H), 5.49 (2H, d, J = 7, CH = CH-CH 2), 5.93 (1H, d, J = 4.5, 7-H) ), 6.28 (1H, d - t, J = 16 & 7, 3 - CH = CH), 7.15 (1H, d, J = 16, 3 - CH) and 8.43 & 9.1 ( each 2H, d, J = 7, Py-H).

51 84830

Example 13. S.

N - j-C-CONH —i-r ^ 5 I J— * © H2N ^^ S N 0 "Y ^ CH ^ H-CH ^ N-n X ° CH3 COO0 H3C —g '

I-1J * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (2-methylthiazolio) -1-propen-1-yl] -3-Cephem-4-carboxylate (I-1J)

To a mixture of diphenylmethyl-7- [2- (5-amino-15,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl yl) -3-cephem-4-carboxylate (IX-1) (E; 714 mg, 1 mmol) and 2-methylthiazole [prepared according to the method of RP Kurkjy and EV Brown; J. Am. Chem. 74 (1952), pp. 5778] (198 mg, 20 mmol) in dry CH 2 Cl 2 (10 mL), AgBF 4 (90% purity; 217 mg, 1 mmol) was added at -20 ° C. The mixture was stirred ... for 30 minutes at room temperature and filtered. The precipitate was extracted with 10% CH 3 OH / CHCl 3 (3 x 20 mL).

. The combined extracts were washed with brine (2 x 5 ml), dried over MgSO 4 and evaporated to dryness to give a yellow residue which was triturated with isopropyl ether and filtered to give 350 mg of quaternized product. A mixture of this solid, sodium hydrogen sulfite (35 mg) and formic acid (3.5-30 ml) was stirred at 40 ° C for 30 minutes. The mixture was concentrated to remove formic acid and the residue was diluted with water (40 mL). A small amount of insoluble matter was removed by filtration. The filtrate was applied to a reversed phase column * ... * (PrepPAK-500 / C18, 100 mL). The column was eluted sequentially with water (200 mL), 5% aqueous CH 3 OH (400 mL), 52 84 8 30, and 10% aqueous CH 3 OH (300 mL). fractions were combined by HPLC analysis (Lichrosorb RP-18; 4 x 300 mm, 0.01 M ammonium sulfate buffer, pH 7.2 with 20% CH 3 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: KBr cm-1 3300, 1760, 1660 and 1600.

UV; Aeake. (phosphate buffer, pH 7) nm (E1 * ^ ,,,) 238 (442) and 10 292 (421).

NMR: D 2 O + DMSO-d 6 δ ppm 3.06 (3H, s, thiazole-CH 3), 3.74 (2H, br - s, 2-H), 4.19 (3H, s, OCH 3), δ, 92 (1H, d, J * 4.5, 7-H), 6.1 (1H, m, 3-CH = CH), 6.8 (1H, d, J = 16, 3-CH trans) and 8.04 & 8.23 (1H, d, J = 4, thiazole-H each). 15 Example 14 N ---- C-CONH - | -f S ^ u N ^ s'N \ - ^ ch = ch-ch2 ^ ch2oh 20 0CH3 C00®

I-1L * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-; Iminoacetamido] -3- [3- (4-hydroxymethylpyridinio) -1'-propen-1-yl] -3-cephem-4-carboxylate (I-1L) • A mixture of diphenyl-7- [2 - (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate ( IX-1) (E-isomer; 1.07 g, 1.5 mmol), 4-hydroxymethylpyridine (818 mg, 7.5 mmol) in CH 3 CN (4.5 mL) and CH 3 OH (3 ml), stirred at room temperature under N 2 for 1 hour. After removal of the solvents by evaporation, the residual oil was triturated with isopropyl ether, the precipitate was collected by filtration and washed with a mixture of isopropyl ether and methanol (53: 8430) (3: 1; 10 ml) to give 1.28 g of 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 N2 for 1 hour. After the addition of 85% HCOOH (5 mL), the mixture was stirred under the same conditions for an additional hour. Toluene was added and the reaction mixture was evaporated to dryness azeotropically in vacuo. The residue was triturated with acetone to give 1.17 g of crude formate of the title compound. A suspension of this compound (1.15 g) in water (100 mL) was filtered to remove insoluble material, which was washed with water (2 x 10 mL). The filtrate and washings were combined and chromatographed on a reverse phase column. 15 A column packed with a packing that had been taken

From a PrepPAK-500 / C18 cartridge column (Waters) (60 mL), was developed sequentially with water, 5% methanol, and 10% methanol. Fractions containing the desired compound were combined, concentrated in vacuo and precipitated by the addition of acetone to give 100 mg of the title compound (I-1L) as a pale yellow powder.

To a suspension of the powder (90 mg) in methanol (9 mL) was added 1 M HCl in CH 3 OH (0.5 mL) and the mixture was stirred at room temperature and concentrated in vacuo. Isopropanol was added to the concentrate to give 77 mg of the hydrochloride of the title compound. Pale yellow powder. Sp. > 190 ° C (dec.).

IR: KBr cm-1 V3 ^ 1775, 1670, 1635 and 1530.

UV: ^ - payment. " (phosphate buffer, pH 7) nm (ε), 230 (22,600) and 30,264 (incl. 16,300).

NMR: D 2 O δ ppm 3.83 (2H, br, 2-CH), 4.17 (3H, s, OCH 3, 5.06 (2H, s, O CH 2 OH), 5.36 (1H, d, J * 4.5 Hz, 6-H), 5.41: -: 35 (2H, d, J = 7 Hz, CH 2 CH-CH 2), 5.94 (1H, d, J = 4.5 Hz, 54 84830 7-H), 6.36 (1H, d - t, J = 16 and 7 Hz, CH = CHCH 2), 7.13 (1H, d, J = 16 Hz, CH = CH-CH 2), 8, 08 and 8.83 (2H, d, J = 7 Hz, Py-H each).

Example 15 5 N - ^ - C-CONH —r> XOC „Hc '© \ = /

2 5 COO

10 I-2H * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-ethoxyiminoacetamido] -3- [3- (4-carbamoylpyridinium) -1-propen-1-yl] - 3-Cephem-4-carboxylate (I-2H) To a solution of 200 mg of 7-amino-3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3-cephem-4 carboxylate hydrochloride (E-isomer) in 5 ml of a 50% aqueous acetone solution, 190 mg of 2-ethoxyimino-2- (5-amino-1,2,4-thiadiazole-3- yl) acetyl chloride-20 hydrochloride [prepared according to the method described in published Japanese Patent Application JP (Kokai 57-24 389 (August 2, 1982)] and the pH of the mixture was adjusted to 6.5-7.0 with 2 N Na 2 CO 3 (about 1 The reaction mixture was stirred at 10 ° C for 1 h, acidified to pH 2 with 1 N HCl, and concentrated on an HP-20 column eluting sequentially with 500 mL of water and 25% isopropanol. The fractions containing the desired product were combined and evaporated to dryness in vacuo. treated with isopropanol (20 mL) to give 263 mg (93%) of the title compound (I-2H). Sp. 170 ° C (dec.).

To a stirred suspension of 225 mg (0.40 mmol) of the above zwitterion in 10 mL of methanol was added 1 mL of 1 N HCl in methanol and the mixture was stirred and concentrated in vacuo. 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%). Sp. 160 ° C (decomp.). Estimated purity 65%.

IR: KBr cm-1 V> max 3300, 1780, 1680 and 1620.

UV: Amacli (phosphate buffer, pH 7) nm (£), 227 (22,300) and 288 (22,800).

NMR: D 2 O δ ppm 1.44 (3H, t, J = 7 Hz, OCH 2 -CH 3), 3.74 (2H, br, s, 2-H), 4.45 (2H, q, J = 7 Hz , OCH2-CH3), 5.36 (1H, d, 10 J = 4.5 Hz, 6-H), 5.46 (2H, d, J = 7 Hz, 3-CH = CH-CH2), 5 .92 (1H, d, J = 4.5 Hz, 7-H), 6.20 (1H, m, 3-CH = CH), 7.04 (1H, d, J = 16 Hz, 3-CH) = CH), 8.43 (2H, d, J = 7 Hz,

Py-HA) and 9.10 (2H, d, J = 7 Hz, Py-HB).

Example 16 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridinium) -1-propen-1-yl ] -3-cephem-4-carboxylate (I-1H) (E-isomer) This example illustrates the preparation of I-1H via the last few steps of Reaction Schemes Ia or Ib, wherein the intermediate benzhydryl-7- [2- (5-amino) - 1,2,4-Thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridino) -1-propen-1-yl] -3-cephem-4-carboxylate formate (XXVII -1H) is isolated.

25 A. Benzhydryl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl-2-methoxyiminoacetamido] -3- [3- (4-carbamoyl-1-pyridinium) -1-propen-1-yl] -3-cephem-4-: carboxylate formate (E-isomer) (XXVII-1H)

A solution of Aoli compound XII-1H (Y '= 1 *; E-iso-30 mer) (34 g; purity 75%) in a 1/1 mixture of acetone and CH 3 OH, 200 mL) was taken up in Amberlite IRA-140- column (formate form, 340 ml). The column was eluted with the same solvent mixture. The first fraction (1 liter) was evaporated to a volume of about 100 ml and the brown residue was triturated with isopropyl ether (400 ml). The resulting 56 84 830 powder was collected by filtration and dried in vacuo to give 29 g (75% purity by HPLC) of the title compound XXVII-1H (E-isomer) as a brown powder, melting at> 150 ° C (decomposed). ).

Δ IR: KBr cm -1 1B 3300, 1780, 1680, 1630 and 1600.

UV: EtOH Amax nm (E1 * 3 cJ 282 (186).

NMR: acetone-d 6 / CH 3 OH-d 4 (1: 1) δ ppm 4.0 (3H, s, OCH 3), 5.26 (1H, d, J = 4.5 Hz, 6-H), 5.43 (2H, d, J = 7 Hz, CH 2 N *), 5.99 (1H, d, J = 4.5 Hz, 7-H), 6.5 (1H, m, 3-CH = CH), 6 , 92 10 (1H, s, CHPh 2), 7.1 (1H, d, J = 16 Hz, 3-CH), 7.35 (10H, m,

Ph-H), 8.36 (1H, s, HCOO) and 8.46 & 9.12 (each 2H, d, J = 8 Hz, Py-H).

B. 7 - [(5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoyl-1-pyridinyl) -15-propene 1-yl] -3-cephem-4-carboxylate (I-1H) (E-isomer)

A mixture of XXVII-1H (E-isomer; 29 g; 75% purity) and 85% formic acid (290 ml) obtained in Step A was stirred for 2 hours at room temperature. Evaporation of the mixture to dryness 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 dried in vacuo to give 24 g (50% purity by HPLC) of the crude title compound; “25 t. The brown solid was treated twice with 2 N

With HCl (1 liter and 0.5 liters). The aqueous extracts were combined and applied to a column packed with Diaion HP-20 (1.5 liters). The column was washed with water (8 L) and eluted with 30% CH 3 OH. The fraction containing the desired product 30 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% purity) of the title compound (as a zwitterion) as a yellow powder.

To a suspension of this product in CH 3 OH (100 mL) was added 84830 A solution of 1 N HCl in CH 3 OH (55 mL) was adjusted at room temperature and the mixture was stirred for 30 minutes. The resulting clear solution was filtered to remove insoluble material, concentrated to a volume of about 50 mL, 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% purity) of the title compound I-1H (E-isomer) (HCl salt), m.p.> 180 ° C. (decomposing). Pale yellow powder.

Example 17 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridino) -1-propene-1- yl] -3-cephem-4-carboxylate (I-1H) (E-isomer) This example illustrates the preparation of I-1H via the last few steps of Reaction Schemes Ia or Ib without isolating Intermediate XXVII-1H (formate).

A solution of IX-1 (E-isomer; 27.6 g, 38.5 mmol) and isonicotinamide (22.8 g, 187 mmol) in CH 3 CN (120 mL) and CH 3 OH (100 mL) was added. in the mixture, 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 then at 40 ° C for 1 hour under N 2 protection. 30 Mana. The mixture was evaporated to dryness in vacuo. The residual oil was taken up in toluene (50 ml), evaporated to dryness azeotropically and triturated with acetone (400 ml) to give 27.8 g of crude title compound. This material was treated twice with 2 N HCl (1 L 35 and 0.5 L). The acid extracts were combined and applied to a column, 58,84830, containing HP-20 resin (1.5 L). The column was eluted with water (9 L) and 30% methanol (10 L). Fractions containing the desired compound were combined and evaporated to dryness to give a yellow oil which was triturated with acetone (300 ml) to give 9.35 g of the title compound as a double ion.

To a suspension of the product (9.3 g) in CH 3 OH (180 mL) was added 1 N HCl in CH 3 OH (55 mL) to give a clear solution. The solution was concentrated to about 10,100 mL and diluted with isopropanol to give 9.50 g (75% purity) of the title compound I-1H (E-isomer) as its hydrochloride. Pale yellow amorphous powder. Sp. > 195 ° C (dec.).

Example 18 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoylpyridino) -1-propene-1- yl] -3-cephem-4-carboxylate (I-1H) (E-isomer) This example illustrates the preparation of I-1H via the last step of Reaction Scheme le (7-N-acylation).

To an ice-cooled suspension of 7-amino-no-cephem hydrochloride XXII-H (E-isomer; 5.0 g, 12.6 mmol) in 50% aqueous acetone (100 mL) was added 25 small portions of sodium bicarbonate were added. The pH of the mixture was monitored throughout the reaction with a pH meter. To a cold neutralized solution (pH about 7) was added 2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetyl chloride hydrochloride (4.02 g, 15.6 mmol) in small portions over one hour. The pH of the reaction mixture was maintained at 6.8 to 7.5 by the occasional addition of sodium hydrogen carbonate. The reaction was also monitored by TLC. After complete consumption of compound XXII-H, the mixture was acidified to pH 3 by the addition of 2N hydrochloric acid. The mixture was filtered and the filtrate was concentrated in vacuo. The residue was diluted with 59 84 8 30 acetone (400 mL) to separate the precipitate, which was collected by filtration to give 9.59 g of the crude title compound as a pale yellow powder. The estimated purity by HPLC was 40%. A suspension of 5 crude products (9.5 g) in 2N hydrochloric acid (150 ml) was filtered and the filtrate was adsorbed onto a column of HP-20 resin (500 ml). After washing with water (1.5 L), the column was eluted with 29% aqueous isopropyl alcohol and the eluate was collected in 100 ml fractions. The ha-10 fractions were combined, acidified with 2N hydrochloric acid (10 ml) and evaporated to dryness. The residual oil was triturated with isopropyl alcohol (200 ml) and the precipitate was collected by filtration. After drying over phosphorus pentoxide, 5.18 g of the hydrochloride of the title compound I-1H (E-isomer) was obtained as a yellow amorphous powder. Sp. > 190 ° C (dec.). Estimated purity 75%.

Example 19

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

Method 16 6 g of 85% pure hydrochloride was dissolved in 20 ml of water and filtered through a pad of Celite. The tan filtrate (pH 2) was passed through a reversed phase column (packing from a PrepPAK-500 / C18 cartridge; Waters; 120 ml) eluting with water. The eluate was collected in 120 ml fractions by HPLC monitoring [Lich-30 rosorb RP-18 column, 4 x 300 mm; mobile phase, 0.01 M phosphate buffer (pH 7.2) / CH 3 OH = 85:15; detection, UV (254 nm)]. Fractions number 3-5 were combined, concentrated to about 10 mL and blended with acetone (100 mL) to give 3.3 g of the zwitterionic form of I-1H (pale yellow-amorphous powder; estimated purity 95%).

60 84830

To a suspension of 95% pure powder (3.2 g) in CH 2 OH (32 mL) was added 1 N HCl in CH 3 OH (18 mL) and the mixture was stirred at room temperature until a clear solution was obtained. The solution was filtered and the filtrate was concentrated to about 10 mL. Isopropanol (100 mL) was added to the concentrate to separate a pale yellow precipitate which was collected by filtration, washed with isopropanol (5 mL) and dried to give 2.6 g of HCl salt (amorphous powder; estimated purity 95 10%).

The pH of a solution of 95% pure hydrochloride (1 g) in water (4 mL) was adjusted to 6.5 with NaHCO 3 (200 mg) and stirred for 30 minutes. The crystals separated during stirring were collected by filtration to give 710 mg of compound I-1H (in the form of a zwitterion) as pale yellow prisms. Sp. > 185 ° C (dec.). Microanalysis showed it to be a trihydrate.

IR: KBr cm-1 ^ makB 1780, 1695, 1660, 1630 and 1610.

UV; Amax (phosphate buffer, pH 7) nm (ε) 227 (22,000) and 20,290 (23,000).

NMR: DMSO-d 6 + D 2 O δ ppm 3.45 (2H, br, s, 2-H), 3.9 (3H, s, OCH 3), 4.99 (1H, d, J = 4.5 Hz, 6-H), 5.16 (2H, d, J = 7 Hz, CH 2 N +), 5.61 (1H, d, J = 4.5 Hz, 7-H), 5.8 (1H, d - t , J = 16 & 7 Hz, 3-CH = CH), 6.93 (1H, d, J = 16 Hz, 3-CH) and: 25.18 & 8.89 (2H, d, J = 7 Hz, Py-H).

: V Analysis for C 21 H 20 N 8 O 6 S 2 * 3H 2 O: calculated; C 42.14 H 4.38 N 18.72 S 10.71 obtained: C 42.41 H 4.35 N 18.86 S 11.00

Method 2 After crystalline Compound I-1H was obtained by Method 1, the crystalline zwitterionic form of Compound I-1H could be obtained directly from crude I-1H-hydrochloride by seeding a few crystals of pure Compound I-1H.

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

no 84830

The pH of the solution was adjusted to 6.5 with NaHCO 3 (60 mg) and decolorized with charcoal. A few pieces of crystals obtained by Method 1 were added as a seed to the filtrate and stirred overnight at room temperature.

The separated crystals were collected by filtration, washed with water (2 x 2 mL) and dried in vacuo to give 170 mg (80% recovered) of pale yellow prisms of I-1H (zwitterionic form) melting at> 185 ° C (decomposed). ; which compound was identical to the compound obtained by Method 10 (as indicated by IR, UV and NMR values).

The crystalline zwitterionic form of I-1H was slightly water soluble (6 mg / ml in brine at 23 ° C). Example 20

15 S

ii-T-r ° "-rr Ί ^ ch2 ° h 0CH3 C00® X == /

20 I-1K * E

·: 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-hydroxymethylpyridinyl) -1: propenyl] -3-cephem-4-carboxylate (I-1K) *: ·; 25 (E-isomer) A. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-hydroxymethylpyridine) ) -1-propenyl] -3-cephem-4-carboxylate iodide (E-isomer) (XII-1K) To a solution of IX-1 (E-isomer; 1.79 g, 2.5 mmol) In 2.5 mL of CH 3 OH and 7.5 mL of CH 3 CN, 3-hydroxymethylpyridine (545 mg, 5 mmol) was added 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 precipitate formed 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-1K as a tan powder. Sp. 170-180 ° C (dec.).

IR: KBr οη'1 ^, 1780, 1725, 1675, 1615, 1530, 1385, 1225, 1040, 750 and 700.

UV: (C 2 H 5 OH) λmak8 nm (E1 \ cm) 290 (196).

NMR: DMSO + D 2 O δ ppm 3.7 (2H, br, s, 2-H), 3.91 (3H, s, OCH 3), 4.70 (2H, s, Py-CH 2 -OH), δ, 28 (2H, m, CH 2 N +), 5.23 (1H, d, J = 5 Hz, 6-H), 5.90 (1H, de, J = 5 Hz, 7-H), 6.34 ( 1H, m, 3-CH = CH), 6.86 (1H, d, J = 16 Hz, 3-CH), 6.89 (1H, s, CHPh 2), 7.35 (10H, m, Ph- H) and 7.9-8.9 (4H, m, Py-H). B. 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-hydroxymethylpyridinio) -15-propenyl] -3 Cephem-4-carboxylate (I-1K) (E-isomer)

A mixture of XII-1J (E-isomer; 2.0 g, 2.4 mmol) and sodium bisulfite (1 g) in 85% HClOH (10 mL) was stirred at room temperature for 2 h. 20 modes. The reaction mixture was concentrated to about 5 mL in vacuo. 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 a PrepPAK-500 / Cie cartridge (Waters) packing] to give 283 mg (22% ) I-1K as an amorphous powder. The powder was crystallized from 4 N H 2 SO 4 and acetone to give 144 mg of the title compound I-1K as colorless needles. Sp. 30 185-188 ° C (dec.).

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

(phosphate buffer, pH 7) nm (E1, 11cb) 236.5 (283),: 275 (sh) (280) and 292.5 (330).

63 84830 NMR: D 2 O δ ppm 3.75 (2h, s, 2-H), 4.18 (3H, s, OCH 3), 4.97 (2H, s, Py-CH 2 OH), 5.35 (1H, d, J = 4 Hz, 6-H), 5.43 (2H, d, J = 6.5 Hz, CH 2 N +), 5.92 (1H, d, J = 4 Hz, 7-H), 6, 18 (1H, d - t, J = 16 Hz, J = 6.5 Hz, 3-CH = CH-), 6.97 (1H, d, 5 J = 16 Hz, 3-CH), 8.13 (1H, dd, J = 8 Hz, J = 6 Hz, Py-H), 8.60 (1H, d, J = 8 Hz, Py-H), 8.84 (1H, d, J = 6 Hz) , Py-H) and 8.90 (1H, s, Py-H).

Example 21

10 n ~ n ~~ rC0t, HT ~ O

As / 11 N -Ns. ® ft \ H2N b N 0 CH = CH-CH2-N y-CONHCH3

N ° CH3 COO © W

I-1M * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2- (Z) -methoxyiminoacetamido] -3- [3- (4-N-methylcarbamoylpyridine) ) -1-propenyl] -3-cephem-4-carboxylate (I-1M) (E-isomer) A mixture of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazole-3 (yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propenyl) -3-cephem-4-carboxylate (IX-1) (E-isomer; 450 mg, 0.62 mmol) and 4 -N-methylcarbamoylpyridine [prepared according to the method of M. Samejima; Yakugaku Zasshi 80 (1960) p. 1706] (215 mg, 1.58 mmol) in acetonitrile (2 mL) was stirred under nitrogen for 5 hours at room temperature. The mixture was evaporated to dryness in vacuo and the residue triturated with ether to give 530 mg of the quaternary salt.

A mixture of 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 to dryness in vacuo. The residue was triturated with acetone and the crude product was collected by filtration. The crude product was chromatographed on an HP-20-84848 column (1.5 x 18 cm) and the column was eluted with water and 30% aqueous methanol. The methanolic eluate was evaporated to dryness in vacuo and the residue was lyophilized to give 140 mg of an amorphous powder which was further purified (HPLC column; Lichrosorb RP-18, solvent: 15% CH 3 OH) and the HPLC eluate was lyophilized to give 60 mg (18%) of the title product I-1M were obtained. Sp. 180-183 ° C (dec.). Estimated purity 80%. IR: KBr cm-1 max 1760, 1660 and 1600.

UV: λmax (phosphate buffer, pH 7) nm (6) 230 (22,100) and 286 (22,100).

NMR: D 2 O δ ppm 3.08 (3H, s, CONHCH 3), 3.72 (2H, s, 2-H), 4.16 (3H, s, OCH 3), 5.35 (1H, d, J = 4.5 Hz, 6-H), 5.95 (1H, d, J = 4.5 Hz, 7-H), 7.00 (1H, d, J = 16 Hz, 3-CH), .35 (2H, d, J = 6 Hz, pyridine-H) and 9.05 (2H, d, J = 16 Hz, pyridine-H).

Example 22

N rt-C-CONH —j- {S N

20 II N | J_N A * Φ /? - ^

H t) S "N CM = CH-CH 2 -N V COOH

NoCH3 COO® W

I-1N * E / Z = 7/1 25 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4- carboxypyridinio) -1-prop-enyl] -3-cephem-4-carboxylate (I-1N) To a stirred suspension of isonicotinic acid (340 mg, 2.8 mmol) in dry DMF (3.5 mL), Li-30 was adjusted under nitrogen to give N, O-bis (trimethylsilyl) acetamide (0.7 mL, 3.8 mmol). To the resulting clear solution was added in one portion diphenylmethyl 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propenyl ) -3-cephem-4-carboxylate (IX-1) (E-isomer; 720 mg, 1 mmol) and a red solution of 65 84 8 30 were stirred for 1.5 hours at room temperature. The reaction mixture was added dropwise to a stirred saturated sodium chloride solution (50 ml) containing sodium thiophosphate (150 mg). The yellow precipitate was collected by filtration, washed with water and dried to give 722 mg of a pale yellow powder. The powder (700 mg) and sodium hydrogen sulfite (70 mg) were dissolved in 85% formic acid (5 mL) and concentrated. The residue was triturated with acetone (70 ml) and the precipitate was isolated by filtration to give 421 mg of a yellow powder. This crude powder (400 mg) was suspended in water (2 ml), and sodium hydrogen carbonate was added to the suspension. The resulting dark solution was adsorbed onto a packed packed (50 mL) PrepPAK / C18 cartridge (Waters ’System 500) and the column was eluted with water (200 mL). The eluate was divided into 10 fractions (20 ml each) and the desired fractions (fractions 4-7) were combined, acidified to pH 3 with 2N hydrochloric acid and evaporated to dryness. The residue was triturated with acetone (30 ml) and the precipitate was collected by filtration to give 201 mg (37%) of the title compound I-1N as a yellow powder. E / Z 0 7/1; purity 80%. Sp. > 189 ° C (dec.).

IR: KBr cm -1, 1770, 1665 and 1600.

UV: (phosphate buffer, pH 7) nm (&) 227 (22,500) and 25,290 (22,100).

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

66 84830

Example 23

N - C-CONH -T-f S N

5 H2 ^ S ^ 10 oJ- ^ CH: CH-cH ^ f3 0CH3 COO0 I (

I-IO * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (Z) -methoxyiminoacetamido] -3- [3- (2,3-cyclopentenopyridinyl)] ) -1-propenyl] -3-cephem-4-carboxylate (1-10) (E-isomer)

A mixture of diphenylmethyl-7- [2- (5-amino-15,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-

Iodo-1-propenyl) -3-cephem-4-carboxylate (IX-1) (e-isomer; 450 mg, 0.62 mmol) and 2,3-cyclopentenopyridine (217 mg, 1.83 mmol) in acetonitrile (2 ml) was stirred under a nitrogen atmosphere for 4 hours at room temperature. After evaporation in vacuo, the mixture was triturated with ether to give 560 mg of the quaternary salt. A mixture of the solid and 85% 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 to dryness in vacuo and the residue triturated to give 391 mg of crude product, which was purified by chromatography on an HP-20 column (1.5 x 18 cm). The column was eluted with water and 30% aqueous methanol. Evaporation of the methanolic eluate to dryness in vacuo followed by freeze-drying gave 160 mg of an amorphous powder which was further purified by HPLC (column: Lichrosorb; solvent: 10% CH 3 OH). The HPLC eluate was lyophilized to give 50 mg (15%) of the title product 1-10. Sp. > 190 35 ° C (dec.). Estimated purity 75%.

67 84 830 IR: KBr cm-1 max. 1765, 1670 and 1600.

UV: A «aks. (phosphate buffer, pH 7) nm (6-) 235 (20,000) and 283 (25,000).

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

Example 24 10 ^ S.

N-j-j-C-CONH - | -S ^

H2N ^ S ^ \ ° N'V ^ Vs CH = CH-CH? N ^ ~ \ -COOH

? Lp w C2Hc

15 25 I-2N

7- [2- (5-Amino-l, 2,4-thiadiazol-3-yl) -2- (Z) -etok-si] -3- [3- (4-karboksipyridinio) -l-propenyl ] -3-cephem-4-carboxylate (I-2N) (E-isomer) 20 and 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (Z) - Ethoxyiminoacetamido] -3- [3- (4-carboxypyridinio) -1-propenyl] -3-cephem-4-carboxylate (I-2N) (Z-isomer) To a cooled mixture of BSA (1 .0 mL, 4.12 mmol) and isonicotinic acid (506 mg, 4.12 mmol), compound IX-2 (obtained from Preparation No. 21) (1.0 g, 1.37 mmol) was added and the mixture was stirred. protected under nitrogen at room temperature for 2 hours. The mixture was poured into 10% Na 2 S 2 O 3 solution (20 mL) to precipitate 1.3 g of quaternary salt, which was collected by filtration, washed with water and dried. A mixture of solid and sodium hydrogen sulphite (0.3 g) in formic acid (98%; 5 ml) was heated at 40 ° C for 1 hour and evaporated to dryness in vacuo. The residue was triturated with acetone 35 and filtered to give 900 mg of crude product (E-68 8 4 830 propenyl isomer / Z-propenyl isomer = 2: 1). Separation of the isomers was performed by HPLC (column: Lichrosorb; solvent: 15% CH 3 OH). The faster moving HPLC fractions were collected, evaporated to dryness in vacuo and lyophilized to give the E-propenyl isomer of I-2N (44 mg; 6% yield).

The slower moving fractions gave the Z-propenyl isomer of I-2N (32 mg; 4% yield) by the same method.

10 I-2N, E-isomer

Sp. > 200 ° C (decomposed).

IR: KBr cm · 1 ^ max. 1765, 1660, 1620 and 1380.

UV: (water) nm (ε) 228 (2200) and 291 (23,600).

NMR: D 2 O δ ppm 1.45 (3H, t, J = 6 Hz, CH 2 CH 3), 3.72 (2H, S, 2-H), 4.45 (2H, q, CH 2 CH 3), 5.40 ( 1H, d, J = 4 Hz, 6-H), 5.90 (1H, d, J = 4 Hz, 7-H), 7.05 (1H, d, J = 15 Hz, 3-CH), 7.05 (1H, d, J = 15 Hz, 3-CH), m 8.30 (2H, d, J = 6 Hz, Py-H) and 8.95 (2H, d, J = 6 Hz), Pooh).

I-2N, Z-isomer 20 M.p. > 200 ° C (decomposed).

IR: KBr cm * 1 V3 „ak ,. 1765, 1660 (incl.), 1620 and 1370.

UV: λmax (phosphate buffer, pH 7) nm (ε) 225 (22,400) and 275 (sh) (16,000).

NMR: D 2 O δ ppm 1.45 (3H, t, J = 7 Hz, CH 2 CH 3), 3.50 (1H, d, '25 J - 17 Hz, 2-H), 3.75 (1H, d, J = 17 Hz, 2-H), 5.38 (1H, d, J = 4 Hz, 6-H), 5.95 (1H, d, J »4 Hz, 7-H), 6.62 (1H , d, J = 11 Hz, 3-CH-, 8.35 (2H, d, J = 6Hz, Py-H) and 8.92 (2H, d, J = 6Hz, Py-H).

Example 25: 30 N -C-CONH -i-p 2 ch2-ch = ch2 (100 ©

- ••• 35 I-3H * E

69 84 830 7- [2- (5-Amino-1,2,4-thiadlazol-3-yl) -2-propen-3-yloxyimino) acetamido] -3- [3- (4-carbamoylpyridine) ) -1-propenyl] -3-cephem-4-carboxylate (I-3H) (E-isomer) To a solution of 35 mg (0.08 mol) of 7-amino-3- [3- (4-carbamoylpyridine) ) -1- (E) -propenyl] -3-cephem-4-carboxylate hydrochloride in 2 ml of a 50% aqueous acetone solution, 52 mg of 2- [5-amino-1,2,4-thiadiazol-3-yl ] -2- (propen-3-yloxyimino) acetyl chloride dihydro-10 chloride (from Preparation No. 25) and the pH of the mixture was adjusted to 6.5-7.0 with 2 N Na 2 CO 3 solution. The mixture was stirred at room temperature for 1 hour, acidified The residue was chromatographed on an HP-20 resin column eluting with 300 ml of water and 30% CH 3 OH / aqueous solution, and the product-containing fractions were combined and evaporated to dryness in vacuo. was purified on a reverse phase column with a support taken up in Pre pPAK-500 / C18 cartridge (Waters; 30 ml). The column was eluted sequentially with water, 5% CH 3 OH, 10% CH 3 OH and 20% CH 3 OH. The product-containing fractions were combined and lyophilized to give 26 mg (62%) of the title product I-3H. Sp. 160 ° C (dec.), I '·'; IR: KBr cm-1 V> ek, 3400, 1765, 1680, 1605 and 1400.

-: 25 UV; ^ Oake (phosphate buffer, pH 7) nm (€ -) 226 (24,600) and 288 (22,800).

NMR; D 2 O δ ppm 3.75 (2H, s, 2-H), 5.41 (1H, d, J * 5 Hz, 6-H), 5.50 (4H, m, CH 2 N * & CH = CH 2), 5.98 (1H, d, J = 5 Hz, 7-H), 6.20 (1H, m, 3-CH = CH), 7.09 (1H, d, J = 17 Hz, '3- CH), 8.50 (2H, d, J = 7 Hz, Py-H) and 9.16 (2H, d, J <7

Hz, Py-H).

70 84830

Example 26 ^ s \ N p-C-CONH —I-S ^ sKss "I J__N. * @ /? ~ \ 5 h2n d o nch = ch-ch2-n Vconh2 I COO ^

CH 2 C = CH

I-4H * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-propargyloxyiminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-propenyl] -3-cephem-4-carboxylate (I-4H) (E-isomer)

To a solution of 86 mg (0.19 mmol) of 7-amino-3-15 [3- (4-carbamoylpyridinio) -1- (E) -propenyl] -3-cephem-4-carboxylate hydrochloride (XXII-H) 2 In 1% aqueous acetone, 63 mg of 2-propargyloxyimino-2- (5-amino-1,2,4-thiadiazol-3-yl) acetyl chloride hydrochloride (from Preparation No. 26) was added. The pH of the suspension was maintained at 6.5-7.0 with 2 N Na 2 CO 3 solution and then stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 2 with 1 N HCl and evaporated to dryness in vacuo. The residue was diluted with 30 mL of water, neutralized with NaHCO 3 and filtered. The filtrate 25 was transferred to the top of a column packed with a reversed phase support (30 mL) taken from a PrepPAK-500 / C18 cartridge (Waters). The column was eluted sequentially with water, 5% CH 3 OH, 10% CH 3 OH, and 20% CH 3 OH. Fractions containing product were combined and lyophilized to give 13 mg (12%) of the title product I-4H. Estimated purity 70%. Sp. 160 ° C. IR: KBr cm-1 night 3400, 2120, 1765, 1680 and 1610.

UV: \ Bak ·. (phosphate buffer, pH 7) nm (ε) 229 (24,000) and 288 (21,200).

35 NMR: D 2 O δ ppm 3.78 (2H, s, 2-H), 5.15 (2H, d, J = 1 Hz, -CH 2 -C = CH), 5.40 (1H, d, J = 5 Hz, 6-H), 5.50 (2H, m, CH-N *), 5.98 (1H, d, J = 5 Hz, 7-H), 6.20 (1H, m, 3- CH = CH), 71 84830 7.05 (1H, d, J = 17 Hz, 3-CH), 8.50 (2H, d, J = 7 Hz, Py-H) and 9.16 (2H, d , J = 7 Hz, Py-H).

Example 27. s \ 5 N-j- | -C-CONH η-T ^ „2m- ^ s ^ n \ 0 ^ - ch’ch-ch2® / ^ ~ \ -conh2 V coo0

[j I-5H * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-cyclopentyloxyiminoacetamido] -3- [3- (4-carbamoylpyridinyl) -1-propenyl] -3-Cephem-4-carboxylate (I-5H) (E-isomer) To a stirred solution of 139 mg (0.31 mmol) of 7-amino-3- [3- (4-carbamoylpyridinio) -1-propenyl ] - 3-cephem-4-carboxylate hydrochloride in 3.5 ml of a 50% aqueous solution of acetone in an ice-cold bath, 120 mg (0.44 mmol) of 2- (5-amino-1,2,4-20 thiadiazole) were added portionwise. -3-yl) -2-cyclopentyloxyiminoacetyl chloride dihydrochloride (from Preparation No. 27). The pH of the mixture was adjusted to 6.5-7.0 with 2 N Na 2 CO 3 solution (0.9 mL) and stirred for 1 h at 10 ° C. The reaction mixture was acidified to pH 2 with 1 N HCl and evaporated to dryness in vacuo. The residue was chromatographed on an HP-20 resin column (20 mL) and eluted sequentially with 300 mL of water and 30% CH 3 OH / aqueous solution. The product-containing fractions were combined and evaporated to dryness in vacuo. The residue was treated with 60 ml of acetone to give 111 mg (83%) of the title compound I-5H. Sp. 160 ° C (decomp.). Ar damaged purity 70%.

IR: KBr cm -1 3400, 1770, 1680, 1605 and 1530.

UV: max (phosphate buffer, pH 7) nm (ε) 224 (23,300) and 286 (24,600).

728 4 830 NMR: DMSO-d 6 δ ppm 1.70 (8H, br, s, H- (Q), 4.68 (1H, br,

B

s, ^ J), 5.05 (1H, d, J = 5 Hz, 6-H), 5.30 (2H, m, CH 2 N *), 50 5.67 (1H, dd, J = 5 Hz) & 7 Hz, 7-H), 6.20 (1H, m, 3-CH = CH), 7.08 (1H, d, J = 17 Hz, 3-CH), 8.34 (2H, d, J = 7 Hz, Py-H), 9,100 (2H, d, J = 7 Hz, Py-H) and 9.38 (1H, d, J = 7 Hz, 7-NH).

10 Example 28

N-s-C-CONH-N-SNV] CH2COOH

15 OCH3 COO®

I-1P * E

7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-carboxymethylpyridino) -201-propenyl] -3-cephem -4-Carboxylate (I-1P) (E-isomer) A. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-carboxymethylpyridinio) -1-propenyl] -3-cephem-4-carboxylate (XII-1β; iodide) (E-isomer)

To a suspension of 3-carboxymethylpyridine hydrochloride (0.89 g, 5 mmol) in 10 mL of CH 2 Cl 2 was added N, O-bis (trimethylsilyl) acetamide (4.97 mL, 18 mmol) and the mixture was stirred at room temperature. until a clear solution was obtained. Compound IX-1 (1.79 g, 2.5 mmol) was added to the solution and the mixture was allowed to stand at room temperature. After 3 h, 3 mL of CH 3 OH was added to the cooled mixture and the solution was evaporated to dryness in vacuo to give an oil which was triturated with ethyl acetate to give 2.28 g of the title compound XII-1P as a tan powder. . Sp. 161 ° C (dec.).

IR: KBr cm -1 max 1780, 1720, 1675, 1630, 1530, 1385, 1225, 1045, 755 and 700.

5 UV: Aerte. (C 2 H 5 OH) nm (E1% 1 cm) 295 (188).

NMR: DMSO + D 2 O δ ppm 3.70 (2H, br, s, 2-H), 3.90 (5H, s, OCH 3 & Py-CH 2 CO), 5.25 (3H, m, -CH 2 N * & δ -H), 5.92 (1H, d, J = 4.5 Hz, 7-H), 6.35 (1H, m, 3-CH = CH-), 690 (1H, d, J = 16 Hz) , 3-CH = CH-), 6.90 (1H, d, J = 16 Hz, 3-CH), 6.92 (1H, 10 s, CHPh 2), 7.35 (10H, m, Ph-H ) and 8.8-9.0 (4H, m, Py-H).

B. 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (3-carboxymethylpyridinio) -1-propenyl] -3-cephem -4-Carboxylate (I-1P) (E-isomer) A mixture of XII-1P (iodide) (2.28 g) and sodium hydrogen sulfite (1.1 g) in 85% HCOOH (10 ml), stirred at room temperature for 2 hours. The reaction mixture was concentrated to about 5 mL in vacuo. the oily residue was triturated with acetone (100 ml) to give 1.22 g of crude product which was purified by column chromatography (HP-20; 420 ml) to give 533 mg of the title compound I-1P (38%, from compound IX -1) as a pale yellow amorphous powder. Sp. 165 ° C (dec.).

IR: KBr cm-1 V> maka 1770, 1670, 1600, 1530, 1385, 1140 and 1040.

UV: ABak8 (phosphate buffer, pH 6.2) nm (E1 \ CB) 234 (374), 277 (sh) (390) and 290 (402).

NMR: D 2 O + NaHCO 3 δ ppm 3.78 (2H, s, 2-H), 3.92 (2H, s, Py-CH 2 CO), 4.22 (3H, s, OCH 3), 5.40 (1H , d, J - 4 Hz, 6-H), 5.44 (2H, d, J = 6.5 Hz, -CH 2 -N *), 5.97 (1H, d, J = 4 Hz, 7- H), 6.20 (1H, d - t, J = 4 Hz, 7-H), 6.20 (1H, d -, J = 16 & 6.5 Hz, 3-CH = CH), 7 .08 (1H, d, J = 16 Hz, 3-CH), 8.11 (1H, d - d, J = 8 & 7 Hz, Py-H5), 8.53 (1H, d, J 35 = 8 Hz, Py-H4), 8.82 (1H, d, J = 7Hz, Py-H6) and 8.86 (1H, s, Py-H2).

74 84830

Example 29. S \ N —-C-CONH —-r ^

Flat." I „J_. A *

5 H N b N 0 nCH = CH-CH2-N 'Vs-CH2COOH

S ° CH3 COO © W

I-1Q * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-10-iminoacetamido] -3- [3- (4-carboxymethylthiopyridinium) -1-propenyl] -3 -Cephem-4-carboxylate (I-10) (E-isomer) A. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadlazol-3-yl) -2-methoxyaminoacetamido] - 3- [3- (4-carboxyl-15-methylthiopyridinio) -1-propenyl] -3-cephem-4-carboxylate (XXI-10, iodide) (E-isomer) To a suspension of 4-carboxymethylthiopyridine (0, 88 g, 5 mmol) in 10 mL of CH 2 Cl 2, N, O-bis- (trimethylsilyl) acetamide (5 mL, 18 mmol) was added, and the mixture was stirred at room temperature until a clear solution was obtained. Compound IX-1 (E-isomer; 1.79 g, 2.5 mmol) was added to the solution and the mixture was allowed to stand at room temperature. After 3 h, 3 mL of CH 3 OH was added to the cold mixture and the solution was evaporated to dryness in vacuo to give an oily residue which was triturated with ethyl acetate to give 2.43 g of the title compound XII-10 (iodide) as a tan powder.

Sp. 155 ° C (dec.).

IR: KBr cm-1 1780, 1720, 1670, 1625, 1525, 1385, 1225, 3015, 1040, 755 and 700.

UV: λββΙΐΒ (C2H5OH) nm (E1S! LcJ 312 (299).

; NMR: DMSO-d 6 + D 2 O δ ppm 3.70 (2H, br, s, 2-H), 3.93 (3H, s, OCH 3), 5.07 (2H, m, CH 2 -N *), δ , 23 (1H, d, J * 5 Hz, 6-H), 5.90 (1H, d, J = 5 Hz, 7-H), 6.29 (1H, m, 3-CH-CH), Δ 6.87 (1H, d, J = 16 Hz, 3-CH), 6.91 (1H, s, CHPh 2), 7.35 75 84830 (10 H, m, Ph-H) and 7.88 - 8.58 (2H, d, J = 6 Hz, Py-H each).

B. 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carboxymethylthiopyridin-5-nio) -1-propenyl] -3-Cephem-4-carboxylate (I-10) (E-isomer)

A mixture of XXI-10 (iodide; 2.43 g) and sodium hydrogen sulfite (1.1 g) in 85% HCOOH (10 mL) was stirred at room temperature for 2 hours. The reaction mixture was concentrated to about 5 mL in vacuo. The oily residue was triturated with acetone (100 ml), filtered and dried to give the crude product (1.39 g) which was purified by column chromatography (HP-20; 20 ml) to give 577 mg of the title compound 15 I-1Q (39 % of compound IX-1) as a pale yellow amorphous powder. Sp. 188 ° C (dec.).

IR: KBr cm-1> 1765, 1670, 1625, 1530, 1380, 1110 and 1035.

UV: AmakB (phosphate buffer, pH 6.2) nm (E1, (IclI1) 234 (459) 20 and 310 (678).

NMR: D 2 O + NaHCO 3 δ ppm 3.79 (2H, br, s, 2-H), 4.10 (2H, s, S-CH 2), 4.23 (3H, s, OCH 3), 5.25 ( 2H, d, J = 6.5 Hz, CH 2 -N +), 5.39 (1H, d, J * 4.0 Hz, 6-H), 5.97 (1H, d, J = 4 Hz, 7 -H), 6.18 (1H, d - t, J = 15.5 Hz & 6.5 Hz, 3-CH = CH), 7.05 (1H, d, J = 15.5 Hz, 3 -CH) and 7.84 & 8.55 (2H, d, J = 7 Hz, Py-H each).

Example 30. S.

N -j-C-CONH -i-S

As' ”I 0J

H2N & N 0 X CH = CH-CH2-N I

0CH3 C00® I-1A * E / Z = 7/1 76 84 8 30 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (1-methylpyrrolidinio) -1-propenyl] -3-cephem-4-carboxylate sulfate (I-1A; sulfate) 5 A. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazole) - 3-yl) -2-methoxyaminoacetamido] -3- (1-methylpyrrolidinio) -1-propenyl] -3-cephem-4-carboxylate (XII-1A; iodide)

To a cold solution of diphenylmethyl-7- [2-10 (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodopropenyl) -3- cephem-4-carboxylate (IX-1) (from Preparation No. 14) (21.5 g, 30 mmol) in ethyl acetate (300 mL) was added dropwise a solution of 1-methylpyrrolidine (2.55 g, 30 mmol). in ethyl acetate for 15 hours (-5 ml) at -5-0 ° 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-1X; iodide), m.p.> 175 ° C. (decomposing).

IR: KBr cm-1> ak8 3300, 1780, 1730, 1685 and 1615.

UV: λβΛ ,. (C 2 H 5 OH) nm (EI + cm) 218 (435) and 295 (188).

B. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadlazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (1-methyl-pyrrolidino) -1-propenyl ] -3-cephem-4-carboxylate (XXI-1A; chloride)

Compound (XII-1A; iodide) (23 g, 28.7 mmol) was dissolved in a mixture of acetone and methanol (1: 1; 230 mL) and applied to an Amberlite IRA-410 column (chloride form; 230 mL). , which was pretreated with the same solvent mixture. The column was developed as a 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 give 17.9 g (87.7%) of the title compound (XXI-1A; chloride), which melted. At 190 ° C (dec.: 35 ten).

77 84830 IR: KBr cm'1 V> mak> 3380, 1780, 1680 and 1620.

UV: <* max. (C 2 H 5 OH) nm (EI + cm) 220 (369) and 290 (232).

C. 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (1-diethylpyrrolidino) -1-5 propenyl] -3 -Cephem-4-carboxylate sulfate (I-1A; sulfate)

A mixture of compound (XII-1A; chloride) (17.8 g, 25 mmol) in 85% formic acid (178 mL) was stirred at room temperature for 2 hours under a nitrogen atmosphere. The mixture was evaporated to dryness in vacuo and the oily residue was triturated with acetone to give 9.80 g of crude compound I-1A. Evaporation to dryness of the filtrate and acetone washings gave an additional 2.95 g of crude compound I-1A. Two batches of crude material were combined and extracted with 2 N HCl (1 L and 0.5 L). The combined extracts were adsorbed onto Diaion HP-20 resin (1.5 L column) eluting with water and 30% aqueous methanol. The desired fractions were collected and evaporated in vacuo to an oily residue which was triturated successively with isopropanol (200 ml) and acetone (200 ml) to give 7.09 g of a pale yellow powder. This material (6.90 g) was dissolved in water (20 ml) and then chromatographed on a column of PrepPAK-500 / C18 cartridge packing (90 ml). 25 ml of water and 10% aqueous methanol as eluent.

The eluate was collected in 20 ml fractions by HPLC monitoring [column: Nucleocil SSC-ODS-262, 8 x 100 mm; mobile phase: 0.01 m phosphate buffer (pH 7.2) / CH 3 OH 90:10; detection: UV (254 nm)]. Fractions number 4-10 were combined, evaporated to dryness in vacuo and freeze-dried to give 2.28 g of a yellow powder [E / Z --- = 7/1; purity 70% (batch 1)]. Fractions 11 to 85 were worked up in the same manner as described above to give 3.27 g of a yellow powder [E / Z * 5/1; purity 70% (lot 2)]. A portion of batch 1 (1.0 g) was purified by rechromatography on 78,84830 PrepPAK-500 / C18 cartridge pack (90 mL). The column was eluted successively with water and 5% aqueous methanol. The eluate containing the desired compound was evaporated to dryness and lyophilized to give 638 g (E / Z = 7: 1; purity 80%) of a yellow powder. Another portion of batch 1 (1.14 g) was worked up in the same manner to give 880 mg (E / Z = 7/1; purity 80%) of a yellow powder. The two purified samples were combined and a portion (1.45 g) was dissolved in 1 N sulfuric acid (5 mL). The solution was diluted with acetone (315 mL) with stirring. The cream precipitate was collected by filtration to give 1.48 g of the title compound (I-1A; sulfate) (E / Z = 7/1; purity 80%), melting at> 185 ° C (dec).

IR: KBr cm-1 W 3380, 3000, 1765, 1675, 1630, 1535, 1390 and 1115.

UV: ΔBaks (phosphate buffer, pH 7) nm (ε) 236 (19,900) and 291.5 (22,500). 1 H NMR: D 2 O + NaHCO 3 δ ppm 2.36 (4H, br, N), 3.15 (3H, s, 20 J-CH 3 ± N), 3.62 (5H, br, 2 -HN), 3.83 (1H, br, 2-H), -v- 4.13 (2H, d, J = 8 Hz, CH 2 N *), 4.22 (3H, s, OCH 3), δ, 39 (1H, I '' ·· d, J = 4.5 Hz, 6-H), 5.96 (1H, d, J = 4.5 Hz, 7-H), 6.00 (1H, m, 3-CH = CH), 6.67 (1 / 8H, d, J = 10 Hz, 3-CH cis) and 7.04 (7 / 8H, d, J = 16 Hz, 3-CH trans) .

Example 31

N -j-C-CONH - | -f S N

30 n \ N * © HX n CH = CH-CH2-N (CH3) 3 2 \ * »och3 COO ^ I-1 * D / E / Z = 10/1 79 84 330 7- [2- (5-amino- 1,2,4-Thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3-trimethylammonio-1-propenyl] -3-cephem-4-carboxylate (I-1D) A. Diphenylmethyl- 7- [2- (5-Amino-1,2,4-thiadiazol-5-yl) -2-methoxyiminoacetamido] -3- (3-trimethylammonio-1-propenyl] -3-cephem-4 carboxylate (XII-ID; iodide)

To a solution of 13.0 g (19 mmol) of diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-10-iminoacetamido] -3- (3- iodopropenyl) -3-cephem-4-carboxylate (IX-1; from Preparation No. 10) in 38 mL of dry ethyl acetate was added 1.75 mL (19.1 mmol) of a 1.1 N solution of trimethylamine in ethyl acetate - At 5 ° C and the mixture was stirred for 1 hour at -5 ° C. The precipitate formed was filtered off, washed well with CHCl 3 and dried to give 12.5 g (88%) of the title compound (XII-1D) as an iodide.

IR: KBr cm-1 v'7max 3300, 1765, 1720 and 1665.

UV: ^ make. (C 2 H 5 OH) nm (&) 300 (18,400).

B. Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-trimethylammonio-1-propenyl) -3 Cephem-4-carboxylate (XII-1D; chloride)

Iodide (XII-1D; 12.5 g) was dissolved in 60 ml of CH 3 OH-25 / acetone solution (1: 1) and passed through an ion exchange resin column [IRA-410 (Cl '); 125 ml]. The column was eluted with 300 ml of CH 3 OH / acetone solution (1: 1) and the eluate was evaporated to dryness in vacuo and triturated with 300 ml of isopropyl / ether solution to give 10.4 g (91%) of 30 quaternary salt (XII- 1D; chlorides).

IR: KBr cm-1 3300, 1765, 1710 and 1665.

UV: ABaks. (C 2 H 5 OH) nm (ε) 298 (15,100).

mp 8,4830 C. 7- [2- (5-amino-1,2,4-thladiazol-3-yl) -2-methoxyiminoacetamido] -3- [3-trimethylammonio-1-propenyl] - 3-Cephem-4-carboxylate (I-1D; sulfate) (E-isomer) To a solution of 10.4 g (16.9 mmol) of XII-1D (chloride) in 20.8 mL of 85% formic acid, allowed to stand for 3 hours at room temperature and evaporated to dryness in vacuo. The residue was treated with 210 ml of isopropanol and the precipitate was filtered off. The solid (120.1 g) was triturated with 210 ml of water and neutralized with sodium hydrogen carbonate. The suspension was filtered and the filtrate was chromatographed on an HP-20 column (300 mL) eluting sequentially with water (1000 mL), 10% CH 3 OH (200 mL) and 30% CH 3 OH (150 mL). ). Fractions containing the desired product were combined and evaporated to dryness in vacuo. The residue was purified by reverse phase chromatography. The column was packed with packing from a PrepPAK-500 / C18 cartridge (Waters; 200 mL). Elution successively with water (600 ml) and 20% and 30% CH 3 OH (200 ml) and then evaporating the fractions containing the desired product to dryness gave 2.52 g (18%) of the title compound. A solution of the zwitterionic product (1.5 g) in 1 N H 2 SO 4 (5 mL) was added portionwise to 300 mL of acetone and the resulting precipitate was filtered off and dried. The sulfate yield of I-1D was 1.42 g (80%). The E / Z ratio was approximately 10/1 based on HPLC.

IR: KBr cm-1 V> nake 3380, 1765 and 1665.

UV: λ max (phosphate buffer, pH 7) nm (ε) 237 (19,500) and 30,293 (22,400).

NMR: D 2 O δ ppm 3.25 (9H, s, N * -CH 3), 3.94 (2H, s, 2-H), 4.14 (2H, d, J = 7 Hz, CH 2 N *), 4 , 23 (3H, s, O-CH 3), 5.42 (1H, d, J = 4.5 Hz, 6-H), 6.00 (1H, d, J = 4.5 Hz, 7-H) ), 6.23 (1H, d - t, J = 7 & 16 Hz, 3 - CH = CH) and 7.23 (1H, d, J = 16 Hz,.

81 84830

Example 32 N - j-C-CONH - | -f S ^ 5 H2N- ^ S "N O Cli * CH-CH ® / ^> - CONH2 OCH3 COO®

I-1H * E

7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-10-iminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-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 (XXII-H; 397 mg, 1 mmol) and NaHCO 3 (168 mg, 2 mmol) in aqueous DMF (3.5 mL water and 7.5 mL DMF) was added to benzotriazol-1-yl-2- (5-amino-1,2,4- thia-diazol-3-yl) -2-methoxyiminoacetate (479 mg, 1.5 mmol) (from Preparation 28). The mixture was stirred at room temperature for 3.5 hours. The pH of the reaction mixture was adjusted to 3-4 with 3 N HCl and the mixture was 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 pH of the solution was adjusted to 6.8 with NaCO 3, treated with decolorizing carbon, concentrated to about 1 mL, and a few pieces of crystalline Compound I-1H were added as seed. After standing overnight, the crystalline precipitate was collected by filtration to give the title compound I-1H (zwitterionic form). Yield-30 to 83 mg (16%). Sp. > 185 ° C (dec.). The physicochemical values of this product were identical to those of Example 10.

82 84830

Preparation 1

Diphenylmethyl 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-chloromethyl-3-cephem-4-carboxylate (IV-1) 5 To the stirred mixture to a suspension of 2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetic acid (111-1) (2.1 g, 10 mmol) in dry CH 2 Cl 2 (50 mL ), PCl 5 (2.09 g, 10 mmol) was added at -30 ° C and the mixture was stirred for 20 minutes at -15 to -20 ° C. To the above acid chloride solution at -30 ° C was added a solution of diphenylmethyl 7-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (II) (4.5 g, 10 mmol) in CH 2 Cl 2 ( 50 mL) with N, O-bis (trimethylsilyl) acetamide (10 g, 50 mmol). After stirring for 1 h at -10 ° C, the mixture was concentrated to remove CH 2 Cl 2 and diluted with ethyl acetate (200 mL). The mixture was washed successively with 10% aqueous NaHCO 3 (2 x 40 mL), water (2 x 20 mL) and brine (10 mL) and dried over MgSO 4. The solvent was evaporated in vacuo and the resulting oily residue (10 g) was dissolved in CHCl 3 (20 ml) and chromatographed on silica gel (Wako gel C-200; 100 g with 10 ml of 1 / 1.5 M phosphate buffer, pH 7). ) using 1 to 3% CH 3 OH / CHCl 3. Fractions containing the title compound were evaporated to dryness to give 5.7 g (95%) of IV-1 as a yellow amorphous powder. Sp. > 140 ° C (decomposed).

IR: KBr cm-1 3300, 1780, 1720, 1680, 1620.

UV: λmax nm (ε) 245 (1,800) and 280 (9,900).

NMR: DMSO-d 6 δ ppm 3.53 (2H, ABq, 2-H), 3.94 (3H, s, OCH 3), δ 4.42 (2H, s, 3-CH 2), 5.22 ( 1H, d, J = 4.5, 6-H), 5.92 (1H, d - d, J = 4.5 & 6.7-H), 6.93 (1H, s, CHPh2), 7 .36 (10H, m. Ph-H), 8.1 (2H, br - s, NH 2), 9.58 (1H, d, J = 6, 7-NH).

83 8 4 8 3 0

Preparation 2

Diphenylmethyl 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-iodomethyl-3-cephem-4-carboxylate (VI) 5 A mixture of was compound IV-1 from Preparation 1 (5.7 g, 9.5 mmol) and NaI (4.3 g, 29 mmol) in dry acetone (50 mL) was stirred for 5 minutes at room temperature. The mixture was evaporated to dryness in vacuo and the resulting oil was shaken with a mixture of ethyl acetate (100 ml) and water (10 ml). The organic layer was separated and washed successively with 10% (w / v) sodium thiosulfate solution and brine. After drying, the ethyl acetate was removed in vacuo to give 6.1 g (93%) of the title compound (V-1) as a yellow amorphous powder, melting at> 120 ° C (dec).

IR: KBr cm-1 Vnakg 3300, 1780, 1725, 1680 and 1620.

UV: Amax (EtOH) nm (ε) 245 (17,000) and 282 (12,000). NMR: DMSO-d 6 δ ppm 3.72 (ABq, 2-H), 3.94 (3H, s, OCH 3), 4.23 (2H, s, 3-CH 2), 5.21 (1H, d , J = 4.5, 6-H), 5.89 (1H, 20 d - d, J = 4.5 & 6.7-H), 6.94 (1H, s, CHPh 2), 7.35 (10H, m, Ph-H), 8.12 (2H, br - s, NH 2) and 9.65 (1H, d, J = 6, 7-NH).

Preparation 3

Diphenylmethyl 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-triphenylphosphonomethyl-3-cephem-4-carboxylate iodide (VI-1 )

A mixture of compound V-1 from Preparation 2 (690 mg, 1 mmol) and triphenylphosphine (786 mg, 3 mmol) in ethyl acetate (20 mL) was stirred overnight at room temperature. The separated solid was collected, washed with ethyl acetate (2 x 10 mL) and dried to give 950 mg (100% phosphonium iodide VI-1). Sp.

186 ° C (dec.).

35 IR: KBr cm-1 3300, 1780, 1710, 1680 and 1610.

84 84830 UV: Amaka (EtOH) nm (ε) 268 (15,000), 275 (13,000) and 300 (7,300).

NMR: DMSO-d 6 δ ppm 3.52 (2H, br - s, 2H), 3.94 (3H, s, OCH 3), 5.34 (1H, d, J = 4.5, 6-H), 5.9 (1H, m, 7-H), 6.3 (1H, s), 7.3 (10H, m, Ph-H) and 7.8 (15H, m, Ph-H).

Preparation 4

Diphenylmethyl 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3 - [(triphenylphosphoranylidene) methyl] -3-cephem-4-carboxylate -10 ti (VII-1)

A mixture of compound VI-1 from Preparation 3 (952 mg, 1 mmol), Aberlite IRA-410 (0T 'form; 500 mg) and 1 N NaOH (4 mL) in CH 2 Cl 2 (10 mL) , was stirred for 1 hour at room temperature. The mixture was filtered and the separated organic layer was dried over MgSO 4 and evaporated to dryness in vacuo. The resulting oil was triturated with ethyl acetate and the resulting yellow precipitate was collected by filtration to give 740 mg (90%) of the title compound VII-1. Sp. > 180 ° C (decomposed).

IR: KBr cm-1 3400, 1750 and 1630.

UV: Et 2 O (EtOH) nm (&) 268 (12,000), 276 (10,000) and 384 (23,000).

Preparation 5

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-chloro-1-propen-1-yl) -3 Cephem-4-carboxylate (VIII-1)

To a solution of VII-1 from Preparation 4 (6.9 g, 8.4 mmol) was added MgSO 4 (3 g) and .... 40% chlorosertaldehyde (810 mg, 8.4 mmol). ). The mixture was stirred for 1.5 hours at room temperature and then filtered. The filtrate was eluted with silica gel (Wako gel C-200; 100 g with 10 ml of 1 / 1.5 M phosphate buffer) on a column using CHCl 3 and CHCl 3 with 35 CH 3 OH. Fractions containing the desired product (0.5-1% 85 84 S 30 CH 3 OH / CHCl 3) were evaporated to dryness in vacuo to give 1.6 g (30%) of the title compound VIII-1 as a yellow amorphous powder which was a mixture of the Z and E isomers of the chloropropenyl moiety. (Z / E = 2/1, based on NMR-5a).

IR: KBr cm-1 key 3300, 1780, 1725, 1680 and 1620.

UV: cA „kB. (EtOH) nm (E) 240 (20,000) and 286 (12,000). NMR: DMSO-d 6 + D 2 O δ ppm 3.56 & 3.8 (m, 2-H), 3.94 (3H, s, OCH 3), 4.16 (d, J = 7.5, CH 2 Cl 2), 5.26 (1H, d, J = 4.5, 6-H), 5.87 (1H, d, J = 4.5, 7-H), 6.28 (2 / 3H, d, J = 11.3-CH cis-H), 6.72 (1 / 3H, d, J = 16, 3-CH trans-H), 6.81 (2 / 3H, s, CHPh2), 6.92 ( 1 / 3H, s, CHPh 2) and 7.4 (10H, m, Ph-H).

Preparation 6 15 Diphenylmethyl 7-benzylideneamino-3- (triphenylphosphoranylidene) methyl] -3-cephem-4-carboxylate (XVI)

To a solution of diphenylmethyl-7-benzylideneamino-3 - [(trifluorophenylphosphonio) methyl] -3-cephene-20 ml-4-carboxylate iodide (XV) [prepared according to the method disclosed in JP Patent Application (Ko-kai) 56-86187 (July 31, 1981)] (60 g, 70 mmol) in CH 2 Cl 2 (350 mL) was added 1 N NaOH (140 mL) Amberlite IRA-410 (0H "form; 35 g) at 5 ° The mixture was stirred for 25 h at 5 ° C. and filtered, the organic layer was separated, dried over MgSO 4, concentrated to a volume of about 100 mL, and stirred with ethyl acetate (500 mL), and the resulting yellow solid was collected. by filtration and dried in vacuo to give 48 g (94%) of the title compound XVI, melting at 195-198 ° C (with decomposition).

IR: KBr cm-1 1770 and 1620.

86 84330

Preparation 7

Diphenylmethyl 7-benzylideneamino-3- (3-chloro-1-propen-1-yl) -3-cephem-4-carboxylate (XVIII) To a stirred solution of compound XVI 5 from Preparation 6 (2.9 g, 4 mmol) ) In a mixture of CH 2 Cl 2 (40 ml) and water (10 ml), anhydrous chloroacetaldehyde (800 mg) was added at room temperature. An additional 800 mg of chloroacetaldehyde was added in three portions over 1 hour and the mixture was maintained at pH 6-9 by the addition of 1 N NaOH. After 15 minutes, the aqueous layer was removed and the organic layer was dried over MgSO 4. Evaporation of the solvent gave a red oil which was dissolved in a mixture of ethyl acetate and isopropyl ether (1: 2; 80 ml). The solution was washed successively with saturated aqueous NaHCO 3 (10 mL) and water (10 mL). After drying over MgSO 4 and removal of the solvent, 3.3 g of a yellow oil were obtained. A solution of the oil in CH 2 Cl 2 (50 mL) was filtered through silica gel (12 g; Wako gel C-200) with 1 / 1.5 M phosphate buffer (1.2 mL; pH 6.4) and the silica gel was washed with CH 2 Cl 2 (50 mL). The filtrate and washings were combined and evaporated to dryness. The residue was triturated with n-hexane to give 1.7 g (80%) of the title compound (XVII) as a yellow powder. The NMR spectrum showed that the chloropropenyl moiety was in the Z configuration. Sp. > 50 25 ° C (decomposed).

IR: KBr cm'1 s? Baka <3400, 1775, 1720 and 1630.

uv: A Baka. (EtOH) nm (ε) 253 (11,000), 258 (11,000), 265 (10,000), 273 (8,300), 281 (7,000) and 290 (6,300).

NMR: DMSO-d 6 δ ppm 3.63 (2H, br - s, 2H), 4.0 (2H, m, CH2-Cl), 5.42 (2H, m, 6-HS3-CH = CH ), 5.72 (1H, d, J = 4.5, 7-H), 6.27 (1H, d, J = 11, 3-CH), 6.85 (1H, s, CHPh 2) and 7 .33 (10H, m, Ph-H).

Preparation of anhydrous chloroacetaldehyde Anhydrous calcium chloride was added with stirring to a cooled solution of 50% aqueous chloroacetaldehyde (50 ml) as the mixture separated into two layers. Chloroacetaldehyde hydrate layer [R. P. Kurkju and E. V. Brown; J. Amer. Chem. Soc. 74 (1952), p. 5778] (upper layer) was separated and diluted with CHCl 3 (100 mL), stirred with MgSO 4 (20 g), heated to reflux for 5 minutes and filtered. The solvent and water were removed by azeotropic distillation (b.p. 56-64 ° C) [S. Trippett and D. M. Walker; J. Chem. Soc. (1961), p. 1266], and the residue was distilled to give anhydrous chloroacetaldehyde [H. O. House, V. K. Jones and G. A. Frank; J. Org. Chem. 29 (1964), p. 3327], p. 70-82 ° C / 760 mm.

IR: cm-1 film max 1720.

Preparation 8 15 Diphenylmethyl 7-amino-3- (3-chloro-1-propen-1-yl) -3-cephem-4-carboxylate (XVIII)

A solution of compound XVII from Preparation 7 (180 mg, 9.34 mmol) in ethyl acetate (10 mL) was added at 5 ° C to a solution of Girard Reagent T [(carboxy-20-methyl) trimethylammonium chloride hydrazide] (251 mg , 1.5 mmol) in CH 3 OH (10 mL) with acetic acid (0.25 mL). After stirring for 30 minutes at 5 ° C, the mixture was evaporated to dryness to remove CH 3 OH and then ethyl acetate (20 mL) was added. The ethyl acetate solution was washed successively with water (2 x 5 mL), saturated aqueous NaHCO 3 (5 mL) and brine (5 mL) and dried over MgSO 4. Evaporation of the solvent gave 145 mg (97%) of the title compound XVIII (Z-isomer) as a yellow powder. Sp. > 100 ° C (decomposed).

30 IR: KBr cm’1 v> eak. 3400, 1770 and 1720.

UV: A “ake. (EtOH) nm (ε) 252 (3,700), 258 (3,800), 260 (4,000), 274 (4,000) and 285 (4,000).

88 84 830

Preparation 9

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-chloro-1-propen-1-yl) -3- Cephem-4-carboxylate (VIII-1) 5 A mixture of 2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetic acid (III-1) (10.1 g , 50 mmol) and PCl 5 (10.4 g, 50 mmol) in dry CH 2 Cl 2 (100 mL) were stirred at -7 to -15 ° C for 2 h. The clear solution was poured into n-hexane (500 mL) to give a precipitate. The organic layer was discarded by decantation and the remaining solid was triturated with n-hexane (100 mL). The yellow precipitate was collected by filtration and dried in vacuo to give 12.5 g (99%) of acid chloride, melting at 80 ° C (decomposed).

15 IR: Nujol cm'1 v ^ aks 1770.

The acid chloride (25 mg, 0.1 mmol) was added to a stirred solution of XVIII (Z-isomer) from Preparation 8 (44 mg, 0.1 mmol) in dry CH 2 Cl 2 (5 mL) at room temperature. After 30 minutes, the mixture was evaporated to dryness in vacuo and diluted with ethyl acetate (20 mL) and saturated aqueous NaHCO 3 (5 mL). The organic layer was washed with saturated aqueous NaHCO 3 (5 mL), brine (5 mL), 10% HCl (5 mL) and then evaporated to dryness to give the product as a yellow foam. The foam was purified by chromatography on a silica gel column (Wako gel C-200; 1 g with 0.1 ml of 1 / 1.5 M phosphate buffer, pH 6.4) eluting with CH 2 Cl 2 / CH 2 OH (100: 1). to give 31 mg (50%) of the title compound 30 VIII-1 (Z-isomer) as a yellow powder. Sp. > 150 ° C (ha-so).

IR: KBr cm-1 v * mak8 3400, 1775, 1720, 1675 and 1630.

UV: Afflak .. (EtOH) nm (ε) 240 (17,000) and 280 (10,000).

89 84830 NMR: DMSO-d 6 δ ppm 3.6 (2H, m, 2-H), 3.92 (3H, s, O-CH 3), 4.0 (2H, m, CH 2 Cl 2), 5.27 ( 2H, m, 6-H & 3-CH = CH), 5.83 (1H; dd, J = 4.5 & 10, 7-H), 6.25 (1H; d, J = 11, 3- CH), 6.83 (1H, s, CHPh 2), 7.33 (10H, m, Ph-H), 8.0 (2H, br - s, 5 NH 2) and 9.57 (1H, d, J = 10, 7-NH).

Preparation 10

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem -4-Carboxylate (IX-1) A solution of VIII-1 from Preparation 5 (Z / E = 2/1; 480 mg, 0.77 mmol) in dry acetone (10 mL) with Nai ( 346 mg, 2.3 mmol) was stirred for 30 minutes at ambient temperature. The reaction mixture was evaporated to dryness in vacuo. The resulting oil was partitioned between ethyl acetate (50 mL) and water (10 mL). The upper layer was washed successively with 10% (w / v) aqueous sodium thiosulfate (10 mL) and brine (10 mL) and dried over MgSO 4. Evaporation of the solvent gave 540 mg (98%) of the title compound 20 IX-1 (Z / E = 1/1) as a reddish amorphous solid melting at> 120 ° C (dec).

IR: KBr cm-1 Vmake 3300, 1780, 1720, 1680 and 1620.

UV: Amaks. (EtOH) nm (E) 240 (21,000) and 290 (12,000).

NMR: DMSO + D 2 O δ ppm 3.67 (2H, m, 2-H), 5.29 (1H, d, J = 4.5, 6-H), 5.95 (1H, d, J = 4.5, 7-H), 6.27 (1 / 2H, d, J = 11, 3-CH cis), 6.72 (1 / 2H, d, J = 16, 3-CH trans), 6 , 87 & 6.96 (each 1 / 2H, s, CHPh 2) and 7.4 (10H, m, Ph-H). Preparation 11

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (3-iodo-1-propen-1-yl) -3 -Cephem-4-carboxylate (IX-1) A mixture of VIII-a (Z-isomer from Preparation 9) (5.6 g, 9 mmol) and NaI (4 g, 27 mmol) in dry acetone (100 ml), stirred for 1.5 hours at room temperature. The mixture was evaporated to dryness and the resulting oil was diluted with ethyl acetate (90 ml). The ethyl acetate layer was washed with 10% (w / v) aqueous sodium thiosulfate (10 mL) and water (10 mL). Removal of the dried (MgSO 4) solvent gave a yellow oil which solidified on trituration with isopropyl ether. Filtration of the precipitate gave 4.3 g (67%) of the title compound IX-1 as the E-isomer. Sp. > 165 ° C (decomposed).

IR: KBr cm-1 Vmax 3400, 1780, 1725, 1680 and 1610.

10 UV: Ληβκβ. (EtOH) nm (ε) 240 (18,000) and 297 (11,000).

NMR: DMSO-d 6 + D 2 δ ppm 3.90 (3H, s, OCH 3), 5.25 (1H, m, 6-H), 5.95 (1H, m, 7-H), 6.72 ( d, J = 16, 3-CH trans), 6.96 (1H, s, CH-Ph 2) and 7.4 (10H, m, Ph-H).

Preparation 12 15 Benzhydryl-7-amino-3- [3-chloro-1-propen-1-yl] -3-cephem-4-carboxylate (Z-isomer) (XVIII) Compound XVIII is a general intermediate used in Reaction Schemes Ib and Ie. .

A. Benzhydryl-7-benzylideneamino-3-triphenylphosphonomethyl-3-cephem-4-carboxylate chloride (XV)

To a suspension of benzhydryl-7-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (II hydrochloride) (200 g, 0.44 mol) in CH 2 Cl 2 (940 mL) was added 1 N NaOH at room temperature. (440 ml). The mixture was shaken for 10 minutes and the organic layer was separated. To the organic layer were added MgSO 4 (75 g) and benzaldehyde (51 g, 0.48 mol) and the mixture was allowed to stand for 3 hours. The reaction mixture was filtered and the insoluble material was washed with CH 2 Cl 2 (200 mL). Triphenylphosphine (126 g, 0.48 mol) was added to the combined filtrate and washings. 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 as a pale yellow crystalline powder which was collected by filtration and dried under vacuum. Yield 322 g (96%). Sp. 185-190 ° C (dec.).

IR: KBr cm-1? MacB 1780, 1720 and 1630.

UV: ^ makB. (CH 2 Cl 2) nm (ε) 260 (24,100).

B. Benzhydryl-7-benzylideneamino-3 - [(triphenylphosphoranylidene) methyl] -3-cephem-4-carboxylate (XVI)

A mixture of compound XV (322 g, 0.42 mol) and 5 N Na 2 CO 3 (252 mL) in CH 2 Cl 2 (1.6 L) was stirred vigorously for 10 minutes at room temperature. The organic layer was separated, dried over MgSO 4 and concentrated to a volume of about 500 mL. The concentrate was added to acetone (1 L) with stirring to give a pale yellow crystalline powder which was collected on a filter pad to give 237 g (78%) of Compound XVI, melting at 195-198 ° C (dec).

IR: KBr cm * 1 V> make 1770 and 1620.

UV: Amake. (CH 2 Cl 2) nm (ε) 254 (23,000) and 389 (22,000).

NMR: CDCl 3 δ ppm 2.56 & 3.16 (2H, ABq), 5.00 (1H; d, J = 4 Hz), 5.23 (1H, d, J = 4 Hz), 5.47 (1H, d, J = 22 Hz), 6.95 (1H, s), 7.2-7.8 (30H, m) and 8.55 (1H, s).

C. Benzhydryl-7-amino-3- [chloro-1-propen-1-yl] -3-cephem-4-carboxylate hydrochloride (z-isomer) (XVIII; hydrochloride) To a boiling solution of compound XVI (214 ') g, 0.294 mol) and N, O-bis (trimethylsilyl) acetamide (40 ml, 0.15 mol) in dry CH 2 Cl 2 (2.9 L) were added dropwise with stirring a 50% solution of chlorine. acetaldehyde (93 g, 0.59 mol) in CHCl 3 over 15 minutes. After standing for 30 minutes, the mixture was evaporated to dryness. To the residual oil were added CH 2 Cl 2 (1.5 L), Grignard Reagent T (99 g, 0.59 mol) and 10% aqueous HCl (300 mL) and the mixture was stirred for 1 h at room temperature. The organic layer was washed with water (200 mL) and saturated NaCl solution (200 mL), dried 92,84830

With MgSO 4, treated with charcoal (5 g) and filtered. The filtrate was cooled to -10 ° C and treated with 1 N HCl in CH 3 OH (300 mL). The mixture was stirred for 30 minutes at room temperature and concentrated to about 300 mL. The concentrate was diluted with ethyl acetate (400 ml) and a few crystals of XVIII hydrochloride were added as seed. After 2 h, 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 the hydrochloride, melting at> 185 ° C (dec). Light yellow needles.

IR: KBr cm-1 2830, 1780 and 1720.

UV: λ (EtOH) nm (ε) 286 (8,800).

NMR: DMSO-d 6 δ ppm 3.73 (2H, br, s, 2-H), 3.97 (2H, m, CH 2 Cl 2), 5.22 (1H, d, J = 4.5 Hz, δ -h), 5.37 (1H, d, J = 4.5

Hz, 7-H), 5.77 (1H, m, 3-CH = CH), 6.45 (1H, d, J = 11 Hz, 3-CH), 6.88 (1H, s, CHPh 2) and 7.33 (10H, br, s, Ph-H). Analysis for C 23 H 21 N 2 O 3 SCl »HCl calculated: C 57.87 H 4.65 N 5.87 S 6.72 Cl 14.85 Found: C 57.62 H 4.53 N 5.70 S 6.64 Cl 14.89

Preparation 13

Benzhydryl 7- [2- (5-amino-l, 2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3 - [- chloro-1-propen-l-yl] -3-cephem -4-carboxylate 25 (Z-isomer) (VIII-1)

To a stirred solution of XVIII (Z-isomer) (20 g, 42 mmol) in CH 2 Cl 2 (420 mL) with N, O-bis (trimethylsilyl) acetamide (34 mL, 125 mmol) was added 2- (5 -amino-1,2,4-thiadiazol-3-yl) -2-methoxy-30-iminoacetyl chloride hydrochloride (15.2 g, 59 mmol) in three portions over 40 minutes at -10 to 0 ° C. The mixture was stirred for 30 minutes at 0-5 ° C and evaporated to dryness in vacuo. The residual brown oil was dissolved in ethyl acetate (420 mL) and the solution was washed sequentially with saturated aqueous NaHCO 3 (3 x 15 mL), saturated 93 84 830

Aqueous NaCl (15 mL) and concentrated to a volume of about 50 mL. To the concentrate was added n-heptane (200 ml) to give 28.5 g (90% purity) of the title compound VIII-1 (Z-isomer) as a colorless powder. Sp. > 150 ° C (decomposed).

IR: KBr cm -1 a V> roake 3400, 1780, 1720, 1680 and 1620.

UV: ABake. (EtOH) nm (1) 240 (20,000) and 283 (12,000).

NMR: acetone-d 6 δ ppm 3.6 (2H, m, 2-H), 3.95 (3H, s, OCH 3), 4.0 (2H, m, CH 2 Cl 2), 5.32 (1H, d, J = 4.5 Hz, 6-H), 5.62 (1H, m, 3-CH = CH), 6.03 (1H, d, J = 4.5 Hz, 7-H), 6, 32 (1H, d, J = 11 Hz, 3-CH), 6.87 (1H, s, CHPh 2), 7.33 (10H, br, s, Ph-H).

Preparation 14

Benzhydryl-7- [2- (5-amino-1,2,4-thiadiazol-3-15-yl) -2-methoxyiminoacetamido] -3- [3-Iodo-1-propen-1-yl] -3 Cephem-4-carboxylate (E-isomer) (IX-1)

A mixture of VIII-1 (Z-isomer) (28.5 g; purity 90%) and sodium iodide (19 g) in dry acetone (420 ml) was stirred for 10 minutes at room temperature and allowed to stand at 5 ° C. 2 hours. The mixture was evaporated to dryness in vacuo. Ethyl acetate (420 ml) and 10% (w / v) aqueous sodium thiosulfate solution (30 ml) were added to the residue, and the mixture was shaken. The organic layer was washed with water (30 ml), dried over MgSO 4 and evaporated to a volume of about 50 ml. The concentrate was diluted with n-heptane (200 mL) to give 30.6 g (95% purity) of the title compound IX-1 (E-isomer) as a yellow powder, melting at 30> 120 ° C (dec).

: IR: KBr cm'1 V>, ak. 3400, 1780, 1725, 1680 and 1620.

UV: λββ1ί ,. (EtOH) nm (ε) 306 (15,000).

NMR: acetone-d 6 δ ppm 3.71 (2H, m, 2-H), 3.97 (3H, s, OCH 3), 4.0 (2H, d, J = 8 Hz, CH 2 I), 5.26 (1H, d, J = 4.5: 35 Hz, 6-H), 6.03 (1H, d - d, J = 4.5 & 8 Hz, are converted to doublets, J = 4.5 Hz by D 2 O, 7-H), 6.32 (1H, d - t, J = 15 & 8 Hz, 3-CH = CH), 6.79 (1H, d, J = 15 Hz) , 3-CH), 6.98 (1H, s, CHPh 2), 7.35 (10H, m, Ph-H), 7.63 (2H, br, s, disappeared with D 2 O, NH 2) and 8 .52 (1H, d, J 5 = 8 Hz, disappeared under D 2 O, 7-H).

Preparation 15

Benzhydryl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- [3- (4-carbamoyl-1-pyridinyl) -1- Propen-1-yl] -3-cephem-4-carb-10-oxylate iodide (E-isomer) (XII-1H)

To a suspension of IX-1 (E-isomer) (30.5 g) and isonicotinamide (26 g, 212 mmol) in CH 3 CN (120 mL) was added CH 3 OH (100 mL) until the mixture became bright. The solution was stirred for 2 hours under a nitrogen atmosphere at room temperature and concentrated to about 100 mL in vacuo. The remaining semi-solid was triturated with isopropyl ether (200 mL). The solvent was removed by decantation and the remaining yellow powder was washed with a mixture of isopropyl ether and CH 3 OH (3: 1; 120 mL). The powder was collected by filtration and dried in vacuo to give 36 g (75% purity by HPLC) of the title compound XII. -1H (E-isomer) as a pale yellow powder, melting at> 150 ° C (decomposed).

IR: KBr cm-1 3300, 1780, 1720, 1680 and 1620.

UV: <Aeake (EtOH) nm (E1, Hlai) 282 (170).

NMR: DMSO-d 6 δ ppm 3.72 (2H, m, 2-H), 3.90 (3H, s, OCH 3),

5.25 (3H, m, 6-H & CH 2 N *), 5.9 (1H, d - d, J = 4.5 & 8 Hz, converted to doublets, J = 4.5 Hz with the addition of D 2 O, 7 -H), 6.35 (1H, m, 3-CH = CH), 6.89 (1H, s, CHPh 2), 6.9 (1H, d, J

- 16 Hz, 3-CH), 7.35 (10H, m, Ph-H), 8.06 (1H, br, s, disappeared with D 2 O, NH 2), 8.21 (2H, br, s , disappeared with the addition of D 2 O, NH 2), 8.36 & 9.07 (each 2H, d, J * 6 Hz, Py-H) and 9.57 (1H, d, J = 8 Hz, disappeared with the addition of 35 D 2 O : ta, 7-NH).

95 84830

Preparation 16

Benzhydryl-7-benzylideneamino-3- [3-chloro-1-propen-1-yl] -3-cephem-4-carboxylate (XVIII) (Z-isomer 5) To an ice-cold mixture of crystalline 7-aminocephem intermediate XVIII (Z-isomer) (13.4 g, 28 mmol) and benzaldehyde (3.3 g, 31 mmol) in ethyl acetate (150 mL) were added dropwise over 20 minutes while maintaining 0.5 N sodium hydroxide (56 mL, 28 mmol). the temperature of the reaction mixture was below 10 [deg.] C. The mixture was stirred under cooling for a further 15 minutes and the organic layer was separated and washed with saturated aqueous sodium hydrogen carbonate solution (2 x 100 ml) and dried over magnesium sulphate. The residual oil was dissolved in carbon tetrachloride (50 ml) and re-evaporated to dryness, this procedure was repeated 3 times and the mixture was monitored by reverse phase TLC to detect any starting material (7-amino acid). 20 cephalosporins) to Schiff's base. Removal of the solvent in vacuo gave 16.45 g of the title compound XVII (Z-isomer) as a pale yellow powder (estimated purity 85%); mp. 74 ° C (dec.); which was used without purification in the next step.

25 IR; KBr cm-1 V5 ^ 1780, 1725 and 1635.

UV: ^ „ak8. (CH 2 Cl 2) nm (E 1 CB) 257 (400).

NMR: CDCl 3 δ ppm 6.18 (1H, d, J = 11 Hz).

Preparation 17

Benzhydryl-7-benzylideneamino-3- [3- (4-carbamoyl-30-yl-1-pyridinyl) -1-propen-1-yl] -3-cephem-4-carboxylate iodide (XXI-H) (E-isomer) Chilled To a mixture of 3-chloropropenyl-lycememe compound XVII (Z-isomer) (16.4 g) in acetone (5 mL) was added dropwise over 10 minutes a solution of sodium iodide 96,84830 (6.3) under nitrogen-35 atmosphere. g, 42 mmol) in acetone (30 mL), and the mixture was stirred at room temperature. The reaction was monitored by UV-absorption ratio [E1 * 1CB (255 nm) / Ex * lcin (320 nm)]. When the ratio reached less than 1.30 (45 after 5 minutes), the mixture was diluted with carbon tetrachloride (400 mL) and allowed to stand at room temperature. After the ratio was smoked to a value less than 1.10 (after 3 hours), the mixture was concentrated to half its volume. The concentrate was treated with a small amount of charcoal and diatomaceous earth and filtered. The filter cake was washed with a 1: 1 mixture of methylene chloride and carbon tetrachloride (100 mL). To the combined filtrate solution and washings was added a solution of isonicotinamide (3.5 g, 28.7 mmol) in dimethylformamide (20 mL) and the mixture was evaporated to dryness in vacuo. The concentrate was allowed to stand at room temperature for 1.5 hours and then washed with isopropyl ether (3 x 100 mL). The remaining 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 phosphorus pentoxide in vacuo, 17 g of the title compound XXI-H (E-isomer) were obtained. Yellow amorphous powder. Sp. 150-155 ° C (dec.). Estimated purity by NMR 80%.

IR: KBr cm-1, 1775, 1725, 1690 and 1635.

UV: λ max. (CH 2 Cl 2) nm (EI +) 258 (335) and 298 (255).

NMR: DMSO-d 6 δ ppm 3.4 - 3.8 (2H, br), 5.35 (2H, br), 5.41 (1H, d, J = 4 Hz), 5.73 (1H, d , J = 4 Hz), 6.93 (1H, s), 6.97 (1H, d, J = 16 Hz), 7.3-7.5 (15H, br, s), 8.40 ( 2H, d, J = 6.5 Hz) and 9.15 (2H, d, J = 6.5 Hz).

97 84 830

Preparation 18 7-Amino-3- [3-4-carbamoyl-1-pyridino) -1-propen-1-yl] -3-cephem-4-carboxylate (XXII-H) (E-isomer) 5 To a suspension in which was quaternized cephem compound XXI-H (17 g) in 85% formic acid (25 ml), concentrated hydrochloric acid (5 ml) was added dropwise 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 washings and poured into acetone (1 L) with stirring. The precipitate formed was collected by filtration to give 9.52 g of a crude product stained yellow. 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 collected by filtration, washed with a small amount of methanol (30 ml) and dried to give 7.58 g of the title compound XXII-H (E-isomer) as the hydrochloride. Pale yellow powder. Purity estimated by UV 85%. Sp. 173-188 ° C (dec.).

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

UV: Amak (i (phosphate buffer, pH 7) nm (E1 * lcm) 294 (457). NMR: D2 O + DC1 δ ppm 3.82 (2H, s), 5.17 (1H, d, J = 5 Hz), 5.33 (2H, d, J * 7 Hz), 5.43 (1H, d, J = 5 Hz), 6.37 (1H, d - t, J = 16 & 7 Hz), 7 , 23 (1H, d, J = 16 Hz), 8.34 (2H, d, J = 7 Hz) and 9.00 (2H, d, J = 7 Hz).

Preparation 19 30 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetyl chloride hydrochloride (II-1; acid chloride-1-hydrochloride) A. 2-Cyano-2-methoxy-aminoacetamide

To a stirred mixture of α-cyanoacetamide 35 (252 g, 3 mol) and sodium nitrite (414 g, 6 mol) in water ββ 84830 (600 ml) was added acetic acid (371 ml, 10 mol) at -5-10 ° C. Within 1.5 hours. The mixture was stirred for another 1.5 hours and the pH was adjusted to 8.5 with 6 N NaOH. Dimethyl sulfate (568 mL, 6 mol) was added to the mixture at 15-20 ° C and the mixture was stirred at 45 ° C for 1.5 hours. The reaction mixture was adjusted to pH 8.5 with 6 NaOH and allowed to stand overnight at 5 ° C to separate the precipitate which was collected by filtration, washed with cold water and air dried to give 292 g (77%) of 10. the title compound as brown needles melting at 170-172 ° C.

IR: KBr cm-1 V> make 3400, 3180, 1720 (sh), 1715, 1690, 1615 and 1570.

UV: (H 2 O) nm {f.) 238.5 (8 290) and 268 (sh, 3870).

NMR: DMSO-d 6 δ ppm 4.20 (3H, s, OCH 2), 7.85 (2H, br, NH 2). Analysis for C4H5N3O2:

Calculated: C 37.80 H 3.97 N 33.06

Found: C 37.43 H 3.75 N 32.51 B. 2-Methoxyiminopropane nitrile 20 A stirred mixture of 2-cyano-2-methoxyiminoacetamide (88.9 g, 0.7 mol), sodium chloride ( 70 g) and phosphorus oxychloride (97 ml, 1.05 mol) in dry 1,2-dichloroethane (350 ml) were boiled for 16 hours. The insoluble material was filtered off through a pad of dicalite and washed with dichloroethane. The filtrate and washings were combined and poured into a stirred ice / water mixture (1.5 L) to decompose excess phosphorus oxychloride. The organic phase was washed with 10% NaHCO 3 (500 mL), water (3 x 500 mL) and saturated NaCl solution (500 mL) and dried over MgSO 4. The filtrate was distilled in vacuo to give 61.5 g (81%) of the title compound, boiling at 62 ° C / 24 mmHg (vol., Bp-47-48 ° C / 12 mmHg).

IR: Liquid membrane cm'1 ^ max. 3020, 2960, 2245, 2020, 1530, 1455 and 1080.

35 NMR: CDCl 3 δ ppm: 4.35 (3H, s, OCH 3).

99 8 4 8 3 0 C. 2-Cyano-2-methoxyiminoacetamidinium acetate

To a solution of anunonium chloride (28.4 g, 0.53 mol) in 28% aqueous ammonia (355 mL) and ethanol (180 mL) was added with stirring and dropwise at 5-15-15 ° C over 30 minutes. a solution of 2-methoxyiminopropanedenitrile (58.0 g, 0.53 mol) in ethanol (120 ml). The mixture was stirred at -10 ° C overnight and then at ambient temperature (20-25 ° C) for one day. The reaction mixture was partitioned between water (350 mL) and CH 2 Cl 2 (350 mL), and the aqueous phase was saturated with sodium chloride and extracted again with CH 2 Cl 2 (300 mL). The organic extracts were combined, dried over MgSO 4 and evaporated to dryness in vacuo. The pH of the residual ethyl acetate solution (1.6 L) was adjusted to 3-4 with acetic acid, whereupon the title compound precipitated as crystals which were collected by filtration and washed with ethyl acetate. Yield 67.6 g (69%). Sp. 152-154 ° C (dec.) [Lit., m.p. 150-155 ° C (dec.)].

IR: KBr cm-1: 3160, 2900, 2360, 2235, 2000, 1665, 1555, 2095 and 1415.

UV: (Et 2 H) nm (ε) 243 (8,500), 265 (sh, 5,380), 305 (sh, 1,400).

NMR: DMSO-d 6 δ ppm 1.88 (3H, s, CH 3 COOH), 4.15 (3H, s, OCH 3) and 7.60 (4H, br).

Analysis for C4H6N4O · CH3C00H

Calculated: C 38.71 H 5.41 N 30.09

Found: C 38.71 H 5.59 N 29.51 D. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetonitrile To a suspension of 2-cyano 2-Methoxyiminoacetamidinium acetate (125 g, 0.672 mol) in CH 3 OH (1.25 L) at -10 ° C was added dropwise triethylamine (234 mL, 1.68 mol) followed by Br 2 (41, 6 ml, 0.806 mol) over 20 minutes at -15 to 10 ° C and the mixture was stirred for 20 minutes. A solution of KSCN (78.3 g, 0.806 mol) in CH 3 OH (550 mL) was added dropwise to the mixture at -15 to 100 8 4 8 3 0 -10 ° C over 1 hour. After stirring for 1 h at 0-5 ° C, the mixture was poured into ice / water (12 L) to form a crystalline precipitate which was collected by filtration, washed with water and air dried to give 120 g (98%) of the title compound. . Sp. 263-265 ° C (dec.). The m.p. is about 60 ° C higher than reported in the literature [Japan Kokai 57-158768, published September 30, 1982, Fujisawa (GB Application 3/6/81); mp. 210-215 ° C (ha so)], but the spectral and microanalytical values are consistent with the structure.

IR: KBr cm-1 mac8 3435, 3260, 3120, 2960, 2245, 2020, 1630, 1545, 1455 and 1415.

UV: ABak8 (EtOH) nm (S) 248 (13,300) and 310 (3,470).

NMR: DMSO-d 6 δ ppm 4.21 (3H, s, OCH 3), 8.30 (2H, br, NH 2). Analysis for C5H5N5OS

Calculated: C 32.78 H 2.75 N 38.23 S 17.50

Found: C 32.76 H 2.51 N 38.02 C. E. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetic acid (111-1)

A mixture of 2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetonitrile (18.3 g, 0.1 mol) in 4 N NaOH (250 mL), heated at 50-55 ° C with stirring for 25 hours. The pH of the reaction mixture was adjusted to 1 with H 3 PO 4 and the mixture was washed with ethyl acetate (100 mL), saturated with NaCl and extracted 3 times with a 3: 1 mixture of ethyl acetate and tetrahydrofuran (2 x 300 mL and 1 x 200 mL). The extracts were combined, dried over MgSO 4 and evaporated to dryness in vacuo. The residue was triturated with isopropyl ether to give pale yellow crystals of the title acid. Yield 16.8 g (83%). Sp. 184-185 ° C (dec); Japan Kokai 57-158768, published September 30, 1982, Fujisawa (GB Application 3/6/81); mp. 35 180-182 ° C (dec.)].

οι 84830 IR: KBr cm-1 3460, 3260, 3140, 1725, 1620, 1605 and 1545.

UV: (H 2 O) n® (^) 234 (13,200) and 288 (sh, 3620).

F. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxy-5-iminoacetyl chloride hydrochloride

To a suspension of 2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetic acid (III-1) (40.4 g, 0.2 mol) in dry CH 2 Cl 2 ( 400 mL), PCl 5 (41.6 g, 0.2 mol) was added in one portion at -50 ° C. The mixture was stirred for 4-10 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 over KH in vacuo to give 46.0 g (90%) of the title acid chloride.

15 IR: (Nujol) cm-1 Vmake 1775.

Preparation 20

N -j-C-CONH —I-S S N

ÄS-N I J_ * 20 H2N ö 0 "γ- ^ CH = CHCH2C10R2 COOCH (Ph) 2

VIII-2 * Z

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (Z) -ethoxyiminoacetamido] -3- [3-chloro-1-propenyl] -3 Cephem-4-carboxylate (VIII-2; Z-isomer)

To a mixture of N, O-bis (trimethylsilyl) acet-30-amide (2.3 mL, 9 mmol) and crystalline diphenylmethyl-7-amino-3- [3-chloro-1- (Z) -propene-1- yl] -3-cephem-4-carboxylate hydrochloride (XVIII) [1.338 g, 2.8 mmol (from Preparation 12)] in methylene chloride (10 mL) was added 2- (5-amino-1,2,4-thiadiazole). 3-yl) -2- (Z) -ethoxyiminoacet-35-yl chloride hydrochloride (800 mg, 2.95 mmol) portionwise at 102 8 4 8 3 0 -10 ° C with stirring and the mixture was allowed to stand at 0 ° C 2 hours. The mixture was diluted with ethyl acetate (200 ml), washed with water and evaporated to dryness in vacuo. Trituration of the residue with isopropyl ether gave 5-title product VIII-2 as an amorphous powder. Yield 1.70 g (95%). Sp. > 150 ° C (decomposed).

IR: KBr cm -1 macr 3300, 1780, 1720, 1690, 1380 and 1220.

UV: AmakB (C 2 H 5 OH) nm (ε) 285 (11,000).

NMR: DMSO-d 6 δ ppm 1.26 (3H, t, J = 7 Hz, CH 2 CH 3), 4.25 (2H, 10 q, J = 7 Hz, CH 2 CH 3), 590 (1H, d - d, J = 4 & 8 Hz, 7-H), 6.26 (1H, d, J = 11 Hz, 3-CH), 6.85 (1H, s, CHPh 2) and 9.53 (1H, d, J = 8 Hz, 7-NH).

Preparation 21

15 N-p-C-CONH —- ^ S N

H \ 0J ch = ch-ch2-i ° COOCH (Ph)? C2H5 20 IX-2 * Mixture of E and Z isomers

Diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2- (Z) -ethoxyiminoacetamido] -3- [3-iodo-1-2-propenyl] -3 Cephem-4-carboxylate (IX-2)

A mixture of VIII-2 (1.90 g, 3 mmol) (from Preparation 20) and sodium iodide (1.4 g, 9 mmol) in acetone (20 mL) was stirred for 10 minutes at room temperature and then allowed to stand at 5 °. At C for 30 3 hours. The mixture was evaporated to dryness in vacuo, diluted with ethyl acetate (100 ml), washed with 10% sodium thiosulphate and water and evaporated to dryness in vacuo. Trituration of the residue with isopropyl ether gave 1.82 g (84%) of the title product IX-2 as a light brown amorphous powder.

103 84830 IR: KBr cm-1? Make <3290, 1770, 1720, 1670, 1530, 1370 and 1220.

UV: ^ max. (C 2 H 5 OH) nm (EI + cm) 340 (199).

Preparation 22 ch "n -] - cs' ί ^ o ^ N ^ ch: ch-ch®N ^ cok„ 2 COOCH (Ph) 2 10

XXI is H; iodides * E

Diphenylmethyl 7-benzylenedenylamino-3 - [(E) -3- (4-carbamoylpyridinyl-1-propenyl] -3-cephem-4-15 carboxylate (XXI-H; iodide) (E-isomer) In a cooled solution of 3 Chloropropenylcephem (XVIII, Z-isomer; 42.8 g, 90 mmol) (from Preparation 16) in dry DMF (80 mL) was added in one portion of KI (20 g, 120 mol) and the mixture was stirred. The reaction was monitored by UV-absorption ratio [E1 \ cn (255 ηπΟ / E1 ca (320 nm)], and when the ratio reached less than 1.10 (after 45 minutes), the mixture was diluted to 800 ° C. ml of methylene chloride, treated with activated carbon (4 g) and filtered, the filter cake was washed with 100 ml of CH 2 Cl 2, isonicotinamide (14.64 g) was added to the combined filtrate and washings, and the mixture was evaporated to dryness. The concentrate was kept at room temperature for 1.5 hours and washed with a mixture of toluene and n-heptane 30 (1: 1; 600 ml). the semi-solid was dissolved in CH 2 Cl 2 (100 mL) and the solution was added dropwise with vigorous stirring to ethyl acetate (3 L). After drying over P2 O5 in vacuo, 57.37 g (88%) of the quaternized title product XXI-H as iodide were obtained.

35 Yellow amorphous powder. Sp. 150-155 ° C (dec.).

104 84 830 This product was identical to the product obtained by iodination with Nairn (Preparation 17).

Preparation 23 5 HC1.H2N- ^ N ^ ^ S'CH = CH-CH2-C1 COOCH (Ph) 2 XVIII * z 10

Diphenylmethyl 7-amino-3- (3-chloro-1-propenyl) -3-cephem-4-carboxylate hydrochloride (Z-isomer) (XVIII; hydrochloride) 25% solution of chloroacetaldehyde (69 g) , 0.22 mmol) in CHCl 3 was added in one portion at -10 ° C to a solution of compound XVI (80 g, 0.11 mol) in CH 2 Cl 2 (1.1 L) with N, O- bis (trimethoxysilyl) acetamide (16.2 mL, 0.06 mol), and the mixture was allowed to stand overnight at 5 ° C. The mixture was concentrated to about 0.3 L of 20 mL, diluted with a solvent mixture of ethyl acetate and isopropyl ether (1: 2; 0.6 L), treated with silica gel (Wako gel C-100; 60 g) and filtered through a pad of di-calite. . The filter cake was washed with the same solvent mixture (0.2 L). The combined filtrate and washings were concentrated to about 0.2 liters, treated with Girard Reagent

Account (60 g, 0.26 mol) and 4 N HCl (220 mL) and a few hydrochloride crystals of XVIII were added as seed. After stirring for 3 hours, the formed crystals were collected by filtration, washed with water (0.5 L) and ethyl acetate (0.5 L) and dried in vacuo to give 37 g (70%) of the title compound XVIII (hydrochloride), which melted. > 185 ° C (decomposed). Light yellow needles. This product was identical to the 12 products in the manufacture.

105 84830

Preparation 24 HCl-HjN-1-Ss> Q-S— N CH = CH-CH2-C1 5 COOCH (Ph) 2

XVIII * Z

Diphenylmethyl 7-amino-3- (3-chloro-1-propenyl) -3-cephem-4-carboxylate hydrochloride (Z-isomer) 10 (XVIII; hydrochloride)

To a solution of chloroacetaldehyde (25% solution in CHCl 3; 628 mg, 2 mmol) in CH 2 Cl 2 (10 mL) at 5 ° C was sequentially added N, O-bis (triethylsilyl) acetamide (0.135 mL, 0.5 mmol) and Compound XVI (728 mg, 1 mmol). The mixture was allowed to stand overnight at 5 ° C. The mixture was evaporated to dryness and diluted with a mixture of ethyl acetate and isopropyl ether (1: 2; 10 ml). The insoluble matter was removed by filtration, and the filtrate was concentrated to about 5 ml. The concentrate was treated with 4 N HCl (2 mL), XVIII hydrochloride was added as a seed 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), m.p.> 185 ° C (dec.). . Light yellow needles. This product was identical to the product obtained in Preparation 12.

Preparation 25 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2- (propen-3-30-yloxyimino) acetyl chloride hydrochloride (III-3; acid chloride hydrochloride) A. Methyl 2- (5-t-Butoxycarbonylamino-1,2,4-thia-diazol-3-yl) -2-propen-3-yloxyimino) acetate A mixture of 685 mg (3.37 mmol) of N-propen-3 -; 35 yloxy) phthalimide [prepared by E. Grochosaki and J.

106 84 830

According to Jurczak's method; Synthesis 1976: 682] and 175 mg (3.35 mmol) of hydrazine hydrate in 5 mL of C 2 H 5 OH were stirred for 1 h at room temperature. The precipitate formed was filtered off and the filtrate and washings were combined. To the solution was added 967 mg (3.37 mmol) of methyl 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-oxoacetate and the mixture was allowed to stand for 1 hour at room temperature and then evaporated. dry using a rotary evaporator. The residue was purified by chromatography on silica gel. The column was eluted with n-hexane / ethyl acetate (4: 1) and the product-rich fractions were combined and evaporated to dryness in vacuo. Yield 514 mg (46%). Sp. 83-86 ° C. IR: KBr cm-1 vBak, 3100, 1745, 1710 and 1620.

15 UV: ÄmakB. (C 2 H 5 OH) nm (ε) 223 (9,700) and 242 (10,000).

NMR: CDCl 3 δ ppm 1.55 (9H, s, BOC-H), 4.40 (2H, d, J = 5 Hz, O-CH 2), 5.21 (2H, m, CH 2 = CH), δ , 90 (1H, m, -CH = CH 2) and 9.50 (1H, br, s, NH).

B. 2- (5-t-Butoxycarbonylamino-1,2,4-thiadiazol-20-yl) -2- (propen-3-yloxyimino) acetic acid [I. Silent et al .; J. Antibiotics 36 (1983), p.

1020]

A solution of 770 mg (2.3 mmol) of methyl 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2- (prop-25-en-3-yloxyimino) acetate and 3.5 mL of 2 N NaOH solution (7.0 mmol) in 15 mL of CH 3 OH were boiled for 30 minutes. The reaction mixture was evaporated to dryness in vacuo and diluted with 10 ml of ethyl acetate / water (1: 1). The aqueous layer was separated, acidified to pH 2 with 6 N HCl and extracted with ethyl acetate (2 x 10 mL). The ethyl acetate solution was dried over MgSO 4 and evaporated to dryness on a rotary evaporator to give 596 mg (81%) of the title compound. Sp. 134-135 ° C [Lit. (reference above): m.p. 135-136 ° C].

35 IR: (Nujol) cm-1 3150, 1745, 1720 and 1550.

107 84 830 UV: λ max (C 2 H 5 OH) nm (ε) 223 (11,000) and 242 (11,300). NMR: DMSO-d 6 δ ppm 1.55 (9H, s, BOC-H), 4.77 (2H, d, J = 5 Hz, O-CH 2), 5.22 (2H, m, CH 2 = CH) and 6.0 (1H, m, CH = CH 2).

C. 2- (5-Amino-1,2,4-tladlazol-3-yl) -2- (propen-3-yloxyimino) acetic acid (III-3) [Japan Kokai 57-112 396 (7 / 13/82; Fujisawa); GB Patent Application 7,935,3538 (10/12/79)]

To a solution of 570 mg (1.74 mmol) of 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2- (propen-3-10-yloxyimino) acetic acid 6 in 1 ml of trifluoroacetic acid, was allowed to stand for one hour at ambient temperature. Evaporation to dryness followed by trituration with 30 ml of isopropyl ether gave 376 mg (95%) of the title compound. Sp. 109 ° C (dec.).

IR: (Nujol) cm-1 3180, 1710, 1545 and 1460.

UV: ABakB. (C 2 H 5 OH) nm (ε) 245 (13,500).

NMR: DMSO-d 6 δ ppm 4.77 (2h, D, J = 5 Hz, O-CH 2), 5.20 (2H, m, CH * CH) and 6.0 (1H, m, CH = CH 2) .

D. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2- (propen-3-yloxyimino) acetyl chloride / hydrochloride

A solution of 350 mg (1.54 mmol) of III-3 and 410 mg (197 mmol) of phosphorus pentachloride in dichloromethane (5 mL) was stirred for 1 h at 25 ° C. The reaction mixture was poured into 60 ml of n-hexane and the precipitate was filtered off. Yield 323 mg.

IR: (Nujol) cm-1 V? Bakg 1765.

Preparation 26 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-propargyloxy-iminoacetyl chloride dihydrochloride (III-4; 30 in acid chloride dichlorohydrin) to methyl Methyl 2- (5-t-butoxycarbonylamino-1 , 2,4-thia-diazol-3-yl) -2-propargyloxyiminoacetate A suspension of 870 mg (4.32 mmol) of N-propargyloxyphthalimide [commercially available; 35 Aldrich) and 200 mg ( 4.0 mmol) of hydrazine hydrate in 5,108,84830 ml of ethanol, stirred at 25 ° C for 1 hour and filtered. To the combined filtrate and washings was added 1.0 g (3.86 mmol) of methyl 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-oxoacetate 5 [I. Silent et al .; J. Antibiotics 36 (1983), p. 1020].

The solution was allowed to stand for one hour and evaporated to dryness in vacuo. Purification by chromatography on silica gel followed by evaporation to dryness gave 319 mg (27%) of the title product. Sp. 72-75 ° C.

10 IR: KBr cm-1 3200, 2380, 1745, 1710 and 1610.

UV: Anak. (C 2 H 5 OH) nm (ε) 235 (12,200).

NMR: DMSO-d 6 δ ppm 1.56 (9H, s, BOC-H), 3.55 (1H, t, J = 2 Hz, CsCH), 4.85 (2H, d, J = 2 Hz, - CH 2 -CHCH) and 8.9 1H, br, S, NH).

B. 2- (5-t-Butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-propargyloxyiminoacetic acid A solution of 490 mg (1.4 mmol) of methyl 2- (5-t -butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-prop-argyloxyiminoacetate and 2.2 ml of a 2N aqueous solution of NaOH (4.4 mmol) in 14 ml of CH 3 OH , boiled for 30 minutes. The reaction mixture was concentrated in vacuo, and 10 ml of ethyl acetate / water (1: 1) was added to the solution. The separated aqueous layer was acidified to pH 2 with 6 N HCl and extracted with ethyl acetate (2 x 10 mL). With a Kui-25 dowel of MgSO 4 and then evaporation of the organic layer to dryness, 149 mg (89%) of the title product were obtained. Sp. 135 ° C (dec.).

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

UV: λmax (C2H5OH) nm (&) 233 (11,500).

NMR: DMSO-d 6 δ ppm 1.55 (9H, s, BOC-H9, 3.55 (1H, t, J = 2 Hz, C = CH), 4.89 (2H, d, J = 2 Hz) , CH 2 Cl 2 CH) and 9.0 (1H, s, NH).

109 84830 C. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-propargyloxyiminoacetic acid (III-4) [Japan Kokai 57-112396 (7/13/82; Fujisawa ); GB Patent Application 7,935,538 (10/12/79)] 5 To a solution of 410 mg (1.26 mmol) of 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3- yl) -2-propargyloxyiminoacetic acid in 5 ml of trifluoroacetic acid, allowed to stand for one hour at 25 ° C. Evaporation to dryness followed by trituration of the residue with 25 ml of isopropyl ether gave 204 mg (72%) of the title compound. Sp. 156-158 ° C (dec.).

IR; (Nujol) cm-1 make 3300, 2480, 1730 and 1610.

UV: imakE. (C 2 H 5 OH) nm (r.) 234 (12,000).

NMR: DMSO-d 6 δ ppm 3.52 (1H, t, J = 2 Hz, CeCH), 4.86 (2H, 15 d, J = 2 Hz, CH2-C = CH) and 8.10 (2H, br, s, NH 2).

D. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-propargyloxyiminoacetyl chloride hydrochloride A mixture of 175 mg (0.07 mmol) of III-4 and 182 mg (0 , 88 mmol) of phosphorus pentachloride in dichloro-20 methane (2 ml), was poured into 30 ml of n-hexane and the precipitate was filtered off. Yield 65 mg (34%).

IR: (Nujol) cm-VBaks. 1770.

Preparation 27 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-cyclopentyl-25-hydroxyiminoacetyl chloride hydrochloride (II1-5; acid chloride hydrochloride) A. Methyl 2- (5-t-butoxycarbonylamino) 1,2,4-thiadiazol-3-yl) -2-cyclopentyloxyiminoacetate A suspension of 860 mg (3.7 mmol) of N- (cyclo-30pentyloxy) phthalimide [U.S. Patent 3,971,778 (7/12/76; Glaxo); GB Patent Application 49,255 (10/25/72)] and 185 mg (3.7 mmol) of hydrazine hydrate in 5 ml of C 2 H 5 OH, stirred for 1 hour at ambient temperature and filtered. The filtrate and washings were combined and added to 1.06 g (3.7 mmol) of methyl 2- (5-t-butoxycarbonylamino) 84830 no-1,2,4-thiadiazol-3-yl) -2-oxoacetate. [I. Silent et al .; J. Antibiotics 36 (1983), pp. 1020]. The solution was allowed to stand for one hour at room temperature and evaporated to dryness in vacuo. The residue was purified by chromatography on a silica gel column. Elution with n-hexane / ethyl acetate (4: 1) followed by evaporation to dryness gave the title product. Yield 906 mg (81%). Sp. 115-118 ° C.

IR: KBr cm-1 3200, 1745, 1720 and 1550.

10 UV: A make. (C 2 H 5 OH) nm (^) 217 (1800) and 252 (7,600).

NMR: CDCl 3 δ ppm 1.51 (9H, s, BOC-H), 1.60 8H, br, s,

B

HQ), 3.88 (3H, s, OCH 3), 4.90 (1H, br, s, XJ-8.70 (1H, br, s, NH) .O 15 B. 2- (5-t- Butoxycarbonylamino-1,2,4-thladiazol-3-yl) -2-cyclopentyloxyiminoacetic acid A solution of 500 mg (1.34 mmol) of methyl 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazole) 3-yl) -2-cyclopentyloxyiminoacetate and 2 N NaOH solution (2 mL, 4 mmol) in 15 mL of CH 3 OH were boiled for 30 minutes, the reaction mixture was evaporated to dryness and 10 mL of ethyl acetate was added to the solution. The aqueous layer was separated, acidified to pH 2 with 6 N HCl and extracted with ethyl acetate (2 x 10 mL), the organic layer was washed with brine, dried over MgSO 4 and evaporated to dryness in vacuo. 377 mg (78%) of the title compound were obtained, mp 185 ° C (dec.).

IR: KBr cm * 1 V> eak. 3160, 1720 and 1550.

UV: Aeak, (C 2 H 5 OH) nm (ε) 238 (13,300).

NMR: DMSO δ ppm 1.51 (9H, s, BOC-H), 1.70 (8H, br, s,

B

Η-ΖΠ) and 4.82 (1H, m, VH).

0 '- 111 84830 C. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-cyclopentyloxyiminoacetic acid (III-5; Z-isomer) [Japan Kokai 57-158769 ( 9/30/82; Fujisawa); GB Patent Application 8,107,134 (3/6/81)] 5 To a solution of 348 mg (0.97 mmol) of 2- (5-t-butoxycarbonylamino-1,2,4-thiadiazol-3- yl) -3-cyclopentyloxyiminoacetic acid in 2 ml of trifluoroacetic acid was allowed to stand for one hour at room temperature. The reaction mixture was evaporated to dryness in vacuo. The residue was triturated with 5 mL of isopropyl ether and 10 mL of hexane to give 215 mg (86%) of the title compound. Sp. 162-165 ° C (dec., [Ref. (Ref. Above): mp 160-165 ° C (dec.)].

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

15 UV: Amaks. (C 2 H 5 OH) nm (ε) 238 (13,300).

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

D. 2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-cyclopentyloxy-aminoacetyl chloride hydrochloride A solution of 190 mg (0.74 mmol) of III-5 and 219 mg ( 1.0 mmol) of phosphorus 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 precipitate formed was collected by filtration. Yield 25 122 mg (60%).

IR: (Nujol) cm-1 1760.

Preparation 28

Benzotriazol-1-yl-2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetate A mixture of 1-hydroxybenzotriazole (2.7 g, 20 mmol) and dicyclohexylcarbodi -imide (4.12 g, 20 mmol) in 65 mL of DMF was stirred at room temperature. After 15 minutes, compound III-1 (4.04 g, 20 mmol) was added to the stirred mixture at 0 ° C and stirring was continued for 3 hours. The reaction mixture was combined to remove 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 gray powder. Sp. 189-192 ° C (dec.).

IR: KBr cm * 1)? Maka. 1815, 1620, 1540, 1415, 1090, 1060, 1005, 945, 865 and 740.

UV: Amaka (C 2 H 5 OH) nm (EI 1 cm) 246 (580) and 283 (sh) (228).

Claims (4)

A process for the preparation of therapeutically active cephemic derivatives of the formula 5 N-1-C CONH-j's R1HN / 3 S / \ O NN 2 NCH = CH-CH 2 ^ N = Q 10 or 2 c <xP (i) wherein R 1 is hydrogen or a common amino protecting group; R 2 is an alkyl group containing 1-4 carbon atoms, a cycloalkyl group containing 3-6 carbon atoms, propenyl or propargyl, or -CH 2 -CH = CH 2 or -CH 2 -C 3 CH is a quaternary ammonio group which is a tri (lower alkyl) ammonio, N- (lower alkyl) pyrrolidinio, 2-methylthiazolio or 2-amino-5-thiazolo [4 , 5-c] pyridinio or pyridinio, which is optionally substituted by a (lower alkyl) -, amino, formylamino, carboxy, carboxy, (lower alkyl) thio, hydroxy (lower alkyl) - , amino (lower alkyl), carbamoyl, or N- (lower alkyl) carbamoyl group, or with an alkylene group attached to the 2,3-positions containing from 3 to 5 carbon atoms, and to prepare their non-toxic pharmaceutically acceptable salts and physiologically hydrolyzing esters, characterized in that a) a compound of formula 120 84830 (0, mm N - γ - C-CONH -, - S 'S \ 5. J is II JZJ) BHN ^ Νχ O ^ NY ^ CH = CHCH_Z (VII) O R COOB1 where R 1 is as defined above, B 2 is a common carboxyl O protecting group, B is hydrogen or a common amino protecting group, Z is chlorine, bromine or iodine, and m is zero or one, is reacted with a tertiary amine Q 2 N (or successively an amine RR'NH and a compound of formula R "Z), and if m is 1, is reduced the sulfoxide by ordinary means, and then all ordinary protecting groups are gradually removed, or b) a compound of the formula COO 30 is reacted with an acid of the formula γ-35's, N n 2 OR * or with said acid acylating derivative, wherein R 2 is above 121 84830, B hydrolyzing esters. Process according to claim 1, characterized in that the following compounds are prepared: 1) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetate amido] -3- [3- (trimethylammonio) -1-propen-1-yl] -3-cephem-4-carboxylate, 2) 7- [2- (5-amino-1,2,4-thiadiazole-3) -yl) -2-methoxy-iminoacetamido] -3- [3- (1-methylpyrrolidinio) -1-propen-1-yl] -3-cephera-4-carboxylate, 3) 7- [2- ( 5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3-pyridino-1-propen-1-yl] -3-cephem-4-carboxylate, 4) 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3- (3-aminopyridinio) -1-propene-1 -yl] -3-cephem-4-carboxylate, 5) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [ 3- (3-Formylaminopyridinio) -1-propen-1-yl] -3-ke-fem-4-carboxylate, 6) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) ) -2-Methoxy-iminoacetamido] -3- [3- (3-aminomethylpyridinyl) -1-propen-1-yl] -3-quin-4-carboxylate, 7) 7- [2- ( 5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3- (3-carbamoylpyridinio) 1-propen-1-yl] -3-cephem-4-carboxylate, 8) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido ] -3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 9) 7- [2- (5-amino-1,2,4-thiadiazole) 3-yl) -2-methoxy-iminoacetamido] -3- [3- (2-methylthiazolio) -1-propen-1-yl] -3-cephem-4-carboxylate, 7- [2 - (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-imino-acetamido] -3- [3- (2-amino-5-thiazolo [4,5-c] pyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 11) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-imino - acetamido] -3- [3- (4-hydroxymethylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 12) 7- [2- (5-amino-1,2,4 (thiadiazol-3-yl) -2-methoxy-imino-acetamido] -3- [3- (3-hydroxymethylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 13) 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3- (4-N-methylcarbamoylpyridinio) -1-propen-1-yl ] -3-cephem-4-carboxylate, 14) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-imino-acetamido] -3- [3- ( 2,3-propylenpyridinio) -l-propen-l-yl] -3- Cephem-4-carboxylate, 15) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-ethoxy-imino-acetamido] -3- [3- (4-carbamoylpyridinio) ) -1-propen-1-yl] -3-cephem-4-carboxylate, 16) 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-cyclopentylio] iminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 17) 7- [2- (5-amino-1,2,4-thiadiazole) -3-yl) -2-propenyl-oxy-iminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxylate, 18) 7- [2 - (5-Amino-1,2,4-thiadiazol-3-yl) -2-propargyl-oxyminoacetamido] -3- [3- (4-carbamoylpyridinio) -1-propen-1-yl] -3- cephem-4-carboxylate, 19) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-imino-acetamido] -3- [3- (4-carboxypyridinio) -1-propen-1-yl] -3-quinate-4-carboxylate, 20) 7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-ethoxy] imino-acetamido] -3- [3- (4-carboxypyridinio) -2-propen-1-yl] -3-ke-fem-4-carboxylate, 21) 7- [2- (5-amino-1,2 (4-Thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3- (3-carboxymethylpyridinio) -1-propen-1-yl] -3-cephem-4-carboxy Lat, 22) 7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- [3- (4-carboxymethylthiopyridinio) -1-propene -1-yl] -3-cephem-4-carboxylate, or their non-toxic, pharmaceutically acceptable salt, silicate, hydrate or physiologically hydrolyzing ester. A compound of the formula chο ch-ch-ch 2 Z R COOB * wherein R is hydrogen or a common amino protecting group, R is an alkyl group containing 1-4 carbon atoms, a cycloalkyl group containing 3-6 carbon atoms, propenyl or proparyl. gyl, B1 is hydrogen or a common carboxyl protecting group and Z is chlorine, bromine or iodine; or its salt, hydrate, sulphate or ester. A compound according to claim 3, characterized in that it is diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- (3-Iodo-1-propen-1-yl) -3-cephem-4-carboxylate, diphenylmethyl-7- [2- (5-amino-1,2,4-thiadiazol-3-yl) - 2-ethoxy-iminoacetamido] -3- (3-iodo-1-propen-1-yl) -3-cephem-4-carboxylate, diphenylmethyl-7- [2- (5-amino-1,2, 4-thiadiazol-3-yl) -2-methoxy-iminoacetamido] -3- (3-chloro-1-propen-1-yl) -3-cephem-4-carboxylate or diphenylmethyl-7- [2- (5-Amino-1,2,4-thiadiazol-3-yl) -2-ethoxy-iminoacetamido] -3- (3-chloro-1-propen-1-yl) -3-cephem-4-carboxylate Or its salt, hydrate, soivate or ester.