GB2101117A - New O-substituted oxime derivatives of 2-amino-thiazol-4-yl-2-hydroxyimino acetic acid - Google Patents

Abstract

The invention relates to O-substituted oxime derivatives of 2-amino-thiazol-4-yl-2-hydroxyimino acetic acid of the general formula: <IMAGE> wherein R16 represents a protecting group for the amino radical and R'i represents: a radical -CX-R'13 in which R'13 represents an alkyl radical having from 2 to 4 carbon atoms, an alkoxy group having from 1 to 4 carbon atoms, a phenyl radical or a radical -(CH2)n-NR''2R''3, in which n represents 0 or an interger of from 1 to 4 and R''2 and R''3, which may be the same or different, each represent a hydrogen atom or an alkyl radical with the proviso that both R''2 and R''3 do not both represent hydrogen when n is zero, or R''2 and R''3 together with the intervening nitrogen atom represent a phthalimido, piperidino or morpholino group, and X is oxygen or sulphur; a radical -CX-NH2; a radical -CHR'4-CO2A''', in which R'4 represents a phenyl or cyano radical and A''' represents an easily-cleavable ester group, a radical <IMAGE> a radical -(CH2)n'-R'5, in which n' is an integer of from 1 to 4 and R'5 represents an alkoxy radical having from 1 to 4 carbon atoms, a radical Alk-S(O)ns- in which Alk represents an alkyl radical containing from 1 to 4 carbon atoms and ns is 0, 1 or 2, or an acyl radical having from 2 to 4 carbon atoms; a radical -(CH2)n2-CN in which n2 is 2, 3 or 4; a radical -(CH2)n'-CX'-NH2, in which X' represents a sulphur atom or, when n' is an integer other than 1, an oxygen atom, a radical <IMAGE> in which R'7 represents a methyl or amino radical; a radical <IMAGE> a radical <IMAGE> in which the positive charge is satisfied by an external anion or internally; a radical -(CH2)n'-R5a in which R5a is a halogen atom, or a radical -S-Rar in which Rar represents a phenyl radical or a 5- or 6-membered aromatic heteroatoms selected from sulphur, nitrogen and oxygen, the phenyl and heterocyclic radicals being optionally substituted by one or more radicals selected from amino, nitro and cyano radicals and alkyl radicals having from 1 to 4 carbon atoms; a radical Rb which is a radical -(CH2)n'-R5b in which R5b represents an imidazolyl, morpholinyl or N-alkyl piperazinyl radical, the N-alkyl radical containing from 1 to 4 carbon atoms; a radical -(CH2)n'-N3; or a radical -(CH2)n'-NR18R19 in which R18 represents a group removable by acid hydrolysis or by hydrogenolysis or an alkyl radical having from 1 to 4 carbon atoms and R19 represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, or R18 and R19 together with the intervening nitrogen atom form a phthalimido group. The derivatives are useful in the preparation of certain cephalosphoranic acids as described in Application Serial No. 2,017,702.

Description

1 GB 2 101 117 A 1

SPECIFICATION

New 0-substituted oxime derivatives of 2-amino-thiazol-4-yi-2hydroxyimino acetic acid This invention relates to new 0-substituted oxime derivatives of 2-amino- thiazol-4-yi-2-hydroxyimino acetic 5 acid useful in the preparation of 0- substituted oxime derivatives of 7-(amino-thiazofyi)-acetamido-cep.h-3em- 4-carboxylic acid described and claimed in our co-pending Application No. 79-11275 (Serial No. 2,017,702) from which the present Application is divided.

In particular, the compounds described and claimed in our co-pending Application are the new syn isomer lo 7-(amino-thiazoiyi)-acetamido-ceph3-em-4-carboxylic acid derivatives of the general formula:

NH2 (11) 5 N 0 15 N H 5 N 20 0 B 0 PN Ft CO2A 25 wherein:

a) B,represents a radical R which is:- 0 a radical -C-R' in which X represents a sulphur or 11 X oxygen atom and R' represents:

an alkyl or alkoxy radical having from 1 to 4 carbon atoms, ora phenyl radical R2 ora radical -(CH2),,-N 1.11R3 in which n represents 0 or an integerfrom 1 to 4 and R2 and R3, which maybe the same or different, each 40 represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, or R2 and R3 together with the intervening nitrogen atom form a piperidino, morpholino or phthalimido group; ii) a radical -CH-C02K 1 M4 in which A' represents a hydrogen atom, an alkali metal atom, an equivalent of an alkaline-earth metal atom or of a magnesium atom, an ammonium group, a substituted ammonium group derived from an organic amino base or an easily-cleavable ester group, R4 represents a phenyl, hydroxyethyl or nitrile radical; iii) the y-lactone of the formula -7(? 0 or iv) a radical -(CHAn-R5 in which n' represents an integer of from 1 to 4 and R5 represents: an alkoxy radical containing from 1 to 4 carbon atoms or a radical Alk - S(O),- in which Alk represents an alkyl radical containing from 1 to 4 carbon atoms and ns represents 0, 1 or 2,.136 ora radical N in which R6 and R7, which may be the same or different, each represent a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, or R6 and R7 together with the intervening nitrogen atom form a phthalimido or 1-pyridino group, or, when n' is an integer other than 1, a cyano radical, 2 GB 2 101 117 A ora radical - C-NH2 11 X, 2 in which X' represents a sulphur atom or, when n' is an integer other than 1, an oxygen atom, ora 4-methy]- or4-amino1,3-thiazol-2-yl radical, ora 1,2,3,4-tetrazol-5-yl radical, oran aziclo radical, oran acyl radical having from 2to 4carbon atoms; b) B represents a radical % which is a radical -(CHAn'-R5,, in which R5,, represents a halogen atom, or a radical -S-Ra, in which R,r represents a phenyl radical or a 5- or 6- membered aromatic heterocyclic radical 10 containing from 1 to 4 heteroatoms selected from sulphur, nitrogen and oxygen, the phenyl and heterocyclic radicals being optionally substituted by one or more radicals selected from amino, nitro and cyano radicals and alkyl radicals having from 1 to 4 carbon atoms; or c) B represents a radical Rb which is a radical -(CH2),'-R5b in which R5b represents an imiclazolyl, morpholinyl or N-alkyl piperazinyl radical, the N-alkyl radical containing from 1 to 4 carbon atoms; R, 15 represents a chloro or methoxy radical; oran alkyl, cycloalkyl or alylthio radical having in each case from 1 to 5 carbon atoms; ora radical -CH2-S-R12 in which R12 represents an optionally-substituted heterocyclic radical containing nitrogen, an acyl radical having from 2 to 4 carbon atoms, the 2-oxo-[3H]-thiazolin-4-yl-carbonyl radical or the 3-m ethyl - 1,2-oxazo 1 -5-yl -ca rbo nyl radical; orthe acetoxymethyl or carbamoyloxymethyl radical; 0 11 ora radical -NH-C-Alk; and A represents a hydrogen atom, or an alkali metal atom, an equivalent of an alkaline-earth metal atom orof a 25 magnesium atom, an ammonium group, a substituted ammonium group derived from an organic amino base or an easily-cieavable ester group and acid addition salts of the derivatives of general formula 1' formed with mineral or organic acids.

According to the present invention there are provided compounds of the general formula:

30 N H R 16 (B) 5"- N 35 \-- C 02H N if ORi 40 wherein R1 6 represents a protecting g rou p for the amino radical and Wi represents:

a radical -CX- Wi3 in which Wi3 represents an al kyl radical having f rom 2 to 4 carbon atoms, an alkoxy group having f rom 1 to 4 ca rbon atoms, a phenyl radical or a radical - (CH2),- N W2W3, in which n represents 0 or an integer of from 1 to 4 and W2 and W3, which may be the same or different, each represent a hydrogen 45 atom or an alkyl radical having from 1 to 4 carbon atoms with the proviso that both W2 and W3 do not both represent hydrogen when n is zero, or W2 and W3 together with the intervening nitrogen atom represent a phthalimido, piperidino or morpholino group and X is oxygen or sulphur; a radical -CX-NH2; a radical -CHR'4-C02A"', in which W4 represents a phenyl or cyano radical and A represents an easily-cleavable ester group; a radical 0 55 a radical -(CH2),'-R'5, in which n is an integer of from 1 to 4 and W5 represents an alkoxy radical having 60 from 1 to 4 carbon atoms, a radical Alk-S(O), in which Alk represents an alkyl radical containing from 1 to 4 carbon atoms and ns is 0, 1 or 2, or an acyl radical having from 2 to 4 carbon atoms; a radical -(CH2),,2-CN in which n2 is 2, 3 or 4; 3 GB 2 101 117 A -- 3 a radical -(CH2)r,'-CX'-NH2, in which X' represents a sulphur atom or, when n' is an integer other than 1, an oxygen atom, a radical 5 N jo in which W7 represents a methyl or amino radical; a radical a radical N-N (CH2,)n' -, 11 -<,v - N H a radical -(CH2)n-R5, in which R5,1 is a halogen atom, or a radical -S- Rar in which Rar represents a phenyl radical or a 5- or 6-membered aromatic heterocyclic radical containing from 1 to 4 heteratoms selected from sulphur, nitrogen and oxygen, the phenyl and heterocyclic radicals being optionally substituted by one or more radicals selected from amino, nitro and cyano radials and alkyl radicals having from 1 to 4 carbon atoms; a radical Rb which is a radical -(CH2)n'-R5b in which R5b represents an imidazolyl, morpholinyl or Walkyl piperazinyl radical, the Walkyl radical containing from 1 to 4 carbon atoms; a radical -(CH2)n'-N3; or a radical -(CH2)n-NIR18R19 in which R18 represents a group removable by acid hydrolysis or by hydrogenolysis or an alkyl radical having from 1 to 4 carbon atoms and R19 represents a hydrogen atom or 30 an alkyl radical having from 1 to 4 carbon atoms, or R18 and R19 together with the intervening nitrogen atom form a phthalimido group.

In one preferred aspect the invention provides compounds of the general formula:

NHR16 (XIII) 35 5''N -,,C 0 2 H 1\ 0 in which R16, n' and R5,, are as defined above.

In another preferred aspect the invention provides compounds of the general formula:

\(CH2) n' - R.5c, NHIR N C 0 2H N (X1111) 1\ 0 (CH2)nli - Hcit i in which W16 represents a trityl or chloroacetyl radical, n'i represents 1 or 2 and Hal' represents a bromine or iodine atom.

4 GB 2 101 117 A In yet another preferred aspectthe invention provides compounds of the general formula:

4 NHR 16 (V110 5"N 5 \-- ' C02H N \ 0 10 R b in which R16 and Rb are as defined above.

In the above formula (B) the radical R% maybe inter alia an acyl group containing from 3 to 5 carbon atoms 15 such as a propionyl, butyryl, isobutyryl, valeryi, isovaleryl or tert- valeryl group, as well as a corresponding sulphur derivative. Radical Wi may also be a methoxycarbonyl, ethoxycarbonyl propyloxycarbonyl, isopropyloxycarbonyl, butoxycarbonyl, isobutyloxycarbonyl or tert- butoxycarbonyl group, or a correspond ing sulphur derivative such as the methoxythiocarbonyl group.

Other possibilities for radical R% include the carbamoyl, N-methylcarbarnoyl, N,N-dimethyl-carbarnoyl, 20 aminoacety], dimethylaminoacetyl, methyl am in opropiony], climethylaminopropionyl, aminovaleryl, dimethyla m i nova leryl, N-piperidinocarbony], N-piperidino-acetyl, Npiperidino-propionyl, N-phthalimido carbonyl, N-phthalimido-acetyl, N-phthalimido-propionyl and benzoyl groups, and the acetyimethyl, acetylethy], propionyl methyl, propionylethyl, methoxymethy], methoxyethyl, ethoxyethy], methoxypropyl, methylth io methyl, methylthioethyl, ethy[thiomethyl and ethylthioethyl groups and oxidised forms of the 25 sulphur-containing radicals such as the methyisulphinyimethyl and methyisu 1 phony] methyl groups.

Radical Wi may also represent the methylaminomethyl, dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl, phthalimidomethyl, phthalimidoethy], phthalimidopropyl, th ioca rba moyl methyl, carbamoylethyl, carbamoylpropyl, th ioca rba moyl ethyl, 4-amino-thiazol2-yl-methyl, 4-methyl-thiazol-2-yl- methyl, 1,2,3,4-tetrazol-5-yi-m ethyl and 1,2,3,4-tetrazol-5-yl-ethyl groups.

When R"i represents the radical -(CH2)n'-R5,, the substituent R5a may be a halo group such as fluoro, chloro, bromo or iodo; and Rar may be a phenyl group; a 1,2,3-,1,2,5-,1,2, 4- or 1,3,4-thiadiazolyl group; a 1 H-tetrazolyl group; a 1,3-thiazoly] group; a 1,2,3-, 1,2,4- or 1,3,4triazoly] group; a 1,2,3-, 1,2,4-,1,2,5- or 1,3,4-oxadiazolyl group; a 2-, 3- or 4-pyridinyl group; a 2- or 3-furyl group; a 2- or 3-thienyl group; or a 2- or 3-pyrolyl group. The phenyl and heterocyclic radicals may be unsubstituted or substituted by one or more 35 radicals selected from amino, nitro, methyl, ethyl, propyl, isopropyl, buty], sec-butyl, tert-butyl and nitrile.

In the compounds of the invention the protecting group R16 may be, for example, an alkyl radical having from 1 to 6 carbon atoms, and preferably is a tert-butyl or tert-amyl radical. R16 may also represent an aliphatic acyl group, an aromatic or heterocyclic acyl group or a carbamoyl group. Examples of such acyl groups include alkanoyl groups such as the formyl, acety], propionyl, butyny], isobutynyl, valeryl, isovaleryl, 40 oxalyi, succinyl and pivaloyl radicals; alkoxycarbonyl or cycloalkoxycarbonyl groups such as methoxycar bonyl, ethoxycarbonyl, propoxycarbonyl, 1 -cyclopropylethoxycarbonyi, isopropoxycarbonyl, butoxycar bonyl, tert-butoxycarbonyl, pentoxycarbonyl, tert-pentoxycarbonyl and Inexyloxycarbonyl radicals; the benzoyl, toluoyl, naphthoyi, phthaloyl, mesyi, phenylacetyl and phenylpropionyl radicals; and aryfalkoxycar bonyl groups such as the benzyloxycarbonyl radial.

These acyl groups may be substituted - for example, by a chlorine, bromine, iodine or fluorine atom - and examples of substituted acyl groups are the chloroacetyl, dichloroacety], trichloroacetyl, trifluoroacetyl and bromoacetyl radicals.

The substituent R16 may also represent a lower aralkyl group such as the benzyl, 4-methoxy-benzyl, phenylethy], trityl, 3,4-dimethoxy-benzyl and benzhydry] radicals, a haloalkyl group such as the trichlor oethyl radical, a chlorobenzoyl, para-nitrobenzoyl, para-tert-butyl- benzoyl, phenoxyacetyl, caprylyl, n decanoyl or acryloxyl radical, or a methylcarbamoyl, phenylcarbamoyl or naphthylcarbamoyl group or a corresponding thiocarbamoyl group.

The possibilities for substituent R16 listed above do not constitute an exhaustive list, and it will be clear to those skilled in the art that other protecting groups, particularly those known from peptide chemistry, can 55 also be used.

Among the preferred protecting groups are the formyl, acetyl, ethoxycarbonyl, mesyl, trifluoroacetyl, chloroacetyl and trityl radicals.

The invention is especially concerned with compounds as described above in which substituent R16 in the compounds of general formula VIA - Vim shown below is a trityl, chloroacetyl, tert-pentoxycarbonyl, 60 tert-butoxycarbonyl or benzy[oxycarbonyl radical.

Especially preferred compounds are those of the above formula (B) in which:

(i) R5a represents a bromine or iodine atom or a phenylthio, 2pyridinylthio, 2-amino-1,3,4-thiadiazol-5-ylthio, 1 -methyl-l H-tetrazol-5-ylthio, 2-amino-phenylthio, 5-nitro-2- pyridiny[thio or 3-cya n o-6-m ethyl -2 pyridinylthio group.

GB 2 101 117 A 5 (ii) R"i represents a radical -CO- R' and R' represents an a] kyl radical having from 2 to 4 carbon atoms, a phenyl radical or a radical -(CH2)1-NR"2R"3 in which n is 0 or 1.

R, 4 1 (iii) Wi represents a radical -uti-tuuR 5 and W4 represents a phenyl radical.

(iv) Wi represents the y lactone of the formula 10 0 (v) R% represents a radical -(CH2)n'-R'5 in which n' is 1 or 2 and W5 represents an acetyl radical. (vi) 13% represents a benzoy], phthalimidoacetyl, phthyalimidomethyl or tetrazo 1 -5-yl-m ethyl radical.

As indicated above the compounds of the present invention are useful as intermediates in the preparation of the compounds of formula (1') of Application Serial No. 2,017,702. Thus the Specification of the parent Application describes as part of the earlier invention processes in which, interalia, compounds of the 20 general formulae:

NH R 16 5"'N \-- "I- C 0 2 H N 1 O-C-R il 13 NHR16 X 5,1N -" C 0 2 H NHR16 511. N C02H 0 -C -NH2 11 N H R 16 X C02H N \ O-CH-CO2A Ill 1 R 1 4 N \0 0 \ J (VIA) (WA) (VIB) (VIC) 6 GB 2 101 117 A 6 NH R 16 5) N \-- "-C 02H N \ C) - (CH2)nt R 1 5 NHIRiA S 1/ N N H R 16 N N NHR16 N \-- -"I- C02H NHR16 N -- -'I- C 0 2 H C02H N l\ 0 (CH2),2 - CN , C02H 1\ 0 (CH2)ni-C-NH2 11.

X N 0-(CH2)n N R7 N N'-N \ o- (CH2W NHR16 N N N H R16 ". N (VID) (VIE) (VIF) (VIG) (VIH) 1 H C02H N I.

1 o - (CH2)ni - HaL C02H NI-o(CH2)n (V1 1 H) (Vij) 7 GB 2 101 117 A 7 NHR16 NHR16 N 5 N C02H (VIK) C02H 5 N 1\0- (CH2) n' - N3 N 1\ 0 (VI'L) NHR16 R b 10 N C02H (VIL) 15 N 0-(CH2)ni-NRI8 Rig NHR16 20 (vim) C02H 25 \ 0 1 C H 2) n 1 - 5 - RQ r or functional derivatives thereof are reacted with a compound of the general formula:

H2N::_ N R, 0 C02A' (IX) to obtain, respectively, compounds of general formulae IIIA, Ill'A, 111B, IIIC, IIID, HIE, II1F, IIIG,][]H, 1IN, IIIJ, 111K, HIL, 1WL and 111m as described in the parent Specification.

The various compounds of general formula (B) may be prepared directly or indirectly from compound of the general formula: 45 NHR16 (V) C02H 50 N 1_. OH 55 Thus, for example, the compounds of general formula VIA may be prepared by a process in which a compound of general formula V is treated with a functional derivative of the acid HO-CX-Wi3, and this reaction is preferably carried out underthe preferred conditions described in the parent Specification forthe 60 analogous reaction to prepare compounds of the general formula IIIA.

The compounds of general formula WA may be prepared by a process in which a compound of general formula V is reacted with a compound of formula X=C=NI-11.

The compounds of general formula VIB may be prepared by a process in which a compound of general formula V is reacted with a compound of the general formula Y-CHW4-CO2A in which Y represents a 65 halogen atom, or a sulphate orsuiphonate group.

8 GB 2 101 117 A The compounds of general formula Vic may be prepared by reacting a compound of general formula V with a compound of the general formula 8 V -/-\c 5 k 1 The compounds of general formula VID maybe prepared by reacting a compound of the general formula V 10 with a compound of the general formula Y-(CH2)n,-R'5.

The compounds of general formula VIE may be prepared by reacting a compound of general formula V with a compound of the general formula Y(CHAn2-M The compounds of general formula VIF may be prepared by a process in which a compound of the general 15 formula:

NHR16 N (VIO \-- "" C02H N \ 0-(CH2W -C N is treated either with hydrogen sulphide or, when n' isan integer other than 1, by hydrolysis in the presence of a base, to obtain the corresponding compound of general formula VIF.

The compounds of general formula VIF wherein X' represents a sulphur atom may be treated with a compound of the general formula W7-CO-CH2-Hal to obtain a compound of general formula VIG.

The compounds of general formula W may also be treated with an azide to obtain the corresponding 30 compound of general formula V1H.

As in the preparation of the compounds of general formula VIA, the preferred conditions for the preparation of compounds WA, V113, VIC, VID, VIE, VIF, VIG and VIH are as described in the parent Specification in relation to the analogous compounds of general formulae 1WA, IME3, IIIC, HID, HIE, II1F, IIIG and 111H.

The starting materials of general formula W wherein n' is 2, 3 or 4 are compounds of general formula VIE 35 and may be prepared by the process described hereinbefore. The compounds VII wherein n' is 1 may be prepared by an analogous process in which a compound of general formula V is reacted with a compound of the general formula Y-CH2-CN, and preferably chloroacetonitrile.

The compounds of general formula WH may be prepared as follows:

a product of the general formula:

N H R t.& "IN 45 N 0 H 50 (wherein Re represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms) - which are compound V and its alkyl esters - is treated with a compound of the general formula Hal -(CH2)n-Hal (wherein the substituents Hal may be the same or different) to obtain a compound of the general formula:

9 N H R [6 C 02R C GB 2 101 117 A 9 N (Vi-H) '11\ 0 10 \1 (CH2) ni - H at which, when % represents a hydrogen atom, is a compound WH and, when Re represents an alkyl radical 15 having from 1 to 4 carbon atoms, is treated first with a base and then with an acid to yield a compound WH.

The compound of general formula: Hal-(C1-12),-Hal, in which Hal preferably represents a bromine or iodine atom, is preferably reacted with the compound V' in the presence of a base to neutralize the hydrohalic acid formed. Suitable bases include mineral bases such as sodium or potassium carbonate or bicarbonate, and organic amine bases such as are known to the skilled man.

The saponification of the esters of general formula W'H wherein R. represents an alkyl radical having from 1 to 4 carbon atoms is carried out under typical known conditions for such reactions. It is possible, for example, firstly to react the ester with a base such as sodium hydroxide, potassium hydroxide or barium hydroxide, and then with an acid such as dilute hydrochloric acid, acetic acid orformic acid.

The compounds of general formula VIj may be prepared by a process in which a compound of general 25 formul WH is reacted with pyridine.

The compounds of general formula VIK may be prepared by a process in which a compound of general formula WH is reacted with an azide.

The compounds of general formula VIL may be prepared by a process in which a compound of general formula WH is reacted with an amine of general formula NHIR181319.

The compounds of general formula WL may be prepared by a process in which a compound WH is reacted with imidazole, morpholine or an Walkyl piperazine.

The compounds of general formula Vim may be prepared by a process in which a compound WH is reacted with a compound of general formula Rar-SH.

The preparation of the compounds of general formulae V1J, VIK, VIL, WL and Vim are preferably carried Out 35 under the preferred conditions described in the parent Specification in relation to the preparation of the compounds IN, 111K, HIL, 1WL and 111m.

It is to be understood that the invention includes functional derivatives of the compounds of the invention which may be reacted with a compound of general formula IX in the above- mentioned process more specifically described in the parent Specification. In a preferred method of carrying out the process described 40 in the parent Specification a compound of general formula IX is treated with a functional derivative of one of the compounds of general formulae VIA to Vim and preferred functional derivatives include a halide, a symmetric or mixed anhydride, an amide or an activated ester.

By way of example, the mixed anhydride may be that formed with isobutyl chloroformate. An example of an activated ester is the ester formed with 2,4-dinitrophenol or 1 - hydroxy-benzo-1 -triazole. Preferred halides 45 are the chloride and the bromide.

It is also possible to employ the acid azicle or acid amide.

The anhydride can be formed in situ by the action of N,N'-disubstitutedcarbodiimides such as N,N-dicyclohexylearbodiimide.

It is to be understood also that the whole of the disclosure of the parent Specification is incorporated herein by way of reference. That is to say insofar as the parent Specification includes a detailed description of the various compounds of the present invention, as well as their various substituents, together with their preparation and use, those details are to be considered as part of the present Specification, along with the various generalities and preferences expressed in the parent Specification.

Following is a description byway of example taken from the parent Specification showing various specific 55

Examples relating to the preparation and use of the compounds of the present invention.

The following Examples and Formulations are given only by way of illustration, to show certain preferred aspects of the invention.

Example 1: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2acetoxyimino- acetylaminolceph-3-em4carboxylic acid.

At ambient temperature, a solution of 0.756 g of the syn isomer of diethylammonium 3-acetoxym ethyl -7-2[2-tritylamino-thiazol-4-yil-2-hydroxyimino-acetylamino)-ceph-3-em-4carboxy late, as prepared below in 1 mi of pyridine and 0.5 mi of acetic anhydride was agitated for 30 minutes. 3 mi of water were added, followed by 2N hydrochloric acid until the pH of the whole was 1. The whole was then vacuum-filtered, and the solid 65 GB 2 101 117 A thus obtained was washed with water and dried to give 0.75 g of crude acylated product. In a water bath, at 45'C, a suspension of 0.520 9 of the crude product thus obtained in 1 mi of aqueous formic acid (HC02H/H20 2:1) was agitated for five minutes, after which 0.5 mi of water were added and the agitation in the water bath continued for 10 minutes at 450C. After the subsequent addition of 5 m[ of ethanol, the whole was concentrated under reduced pressure, and the concentrate taken up with diethyl ether. After evaporating to dryness under reduced pressure, washing and drying, 0.304 g of impure product (containing some deacylated derivative) were obtained. The impure product was dissolved in 1 mi of acetic anhydride and 2 drops of pyridine, from which the purified expected product (0.30 g) was precipitated with 10 m[ of diethyl ether and then recovered.

Microanalysis.. C17H1708N5S2: 523.0 Calculated: C% 44.80 Found: 44.6 N.M.R. Spectrum 2.05 p.p.m.: 0Ac 6.75 p.p.m.: H5 of the thiazol.

H% 3.75 N% 14.70 S% 12.26 3.8 14.5 11.7 Preparation of the syn isomer of diethylammonium 3-acetoxymethyl-7[2-(2- tritylaminothiazol-4-yl)-220 hydroxyimino-acetylaminol-ceph-3-em-4- carboxylate. Stage A: The.y isomer of 2-(2-tritylaminothiazol-4-yl)-2-(1-methyl- 1-methoxyethoxyimino)acetic acid.

At ambient temperatu re, 12.9 g of the syn isomer of 2-hydroxyim ino 2-(2tritylaminothiazol-4-yi)acetic acid in 120 CM3 of methylene chloride and 12 CM3 of 2-methoxy propene were agitated for 20 minutes. The whole was concentrated to dryness and after the addition of 60 CM3 of methylene chloride and 12 CM3 of methoxy propene, agitation was continued for 30 minutes. The whole was then concentrated to dryness under reduced pressure to obtain the expected product, which was used in the following stage without further purification.

Stage 8: Thesyn isomer of diethylammonium 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-(1-methyl- 1-methoxy-ethoxyimino) acetylaminol-ceph-3-em-4-carboxylate.

One dissolves the amount of syn isomer of 2-(2-tritylaminothiazol-4-yi)-2(1 -methy]-1 -methoxyethoxyimino)acetic acid obtained by following the procedure described in Stage A after starting with 47.25 9 of 2hydroxyimino-2-(2-tritylaminothiazol-4-y])-acetic acid, in 230 CM3 of methylene chloride. 12.5 g of dicyclohexyl carbodiimide were added, and the whole was agitated for an hour at ambient temperature. The dicyclohexylurea thus formed was separated by vacuum-filtration and rinsed with a little methylene chloride 35 (9.82 g of dicyclohexylurea were formed). To the filtrate (and rinsings) were added a solution of 13.6 g of 7-amino-cephalosporanic acid in 70 cm' of methylene chloride and 14 CM3 of triethylamine, and the whole was agitated for two hours at ambient temperature. After washing in ampoule form with 350 cm 3 of N hydrochloric acid, decanting, washing with water, drying and concentrating to dryness, a residue was obtained, which was dissolved in 100 CM3 of ethyl acetate. Crystallisation was then initiated, and after 30 40 minutes 5.5 g of starting product were recovered by vacuum-filtration. The filtrate remaining was concentrated to dryness, and the formed residue agitated for 30 minutes with 200 cm 3 of isopropyl ether.

After vacuum-filtration and drying 37.35 g of crude product was obtained, which was purified as follows.

The product was dissolved in 148 cm 3 of ethyl acetate, and 5.5 CM3 of diethylamine were added. The product was then precipitated under intense agitation by adding 650 CM3 of diethyl ether, and after vacuum-filtration, washing with diethyl ether and drying 26.35 g of the expected purified product were obtained. To obtain a second yield of product, the filtrate from the vacuum-filtration was concentrated to dryness and the residue formed taken up with 50 cM3 of ether. After drying 2.8 g of product identical to that of the first yield in T.L.C. were obtained. The combined products were used without any further treatment in the following stage.

N.M.R. Spectrum (CDC13) 60 MHz CH3(a) (a) = 1. 54 p. p. m.

0 CH3(a) (b)=3.27 p.p.m. 55 OCH3(b) proton of the thiazol ring: 6.78 p. p. m.

60 Stage C: The syn isomer of diethylammonium 3-acetoxy-methyl-7-[2(2tritylaminothiazol-4-yl)-2hydroxyimino-acetylaminol-ceph-3-em-4-carboxylate.

7.6 g of the syn isomer of diethylammonium 3-acetoxy-methyl-7-(2-[2tritylaminothiazol-4-yll-2-[1 -methyl 1-methoxy-ethoxyiminol-acetylamino)-ceph-3-em-4-carboxylate, prepared in Stage B above, were dissolved in 30 CM3 of acetone and 10CM3 of 2N hydrochloric acid. The whole was agitated for 40 minutes at ambient 65 11 GB 2 101 117 A 11 temperature, after which 20 CM3 of water were added and the acetone was driven off at WC under reduced pressure.25 CM3 of ethyl acetate were added, and after decanting, re-extracting, washing with water, drying and vacuum-filtering, 1 CM3 of diethylamine was added to the filtrate. After crushing and freezing the product formed was separated by vacuum- filtration and washed with ether to give 6 g of pure product.

Analysis: C38H4007N6S2 Calculated: C% 60.30 H% 5.33 N% 11.10 S% 8.47 Found: 60.5 5.7 10.9 8.2 N.M.R. Spectrum (C13C13) 60 MHz 6.63 p.p.m. thiazol proton 7.33 p.p.m. trityl proton Example 2: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)2(phthalimidoacetoxyimino)acetylaminol-ceph-3-em-4-carboxylic acid.

In 4 mi of methylene chloride were put into suspension under agitation 0. 756 g of diethylaminium 3-acetoxymethyl-7-[2-(2-trityl am i no-th lazol-4yl)-2-hyd roxyi m in o-acetyla m i no]-cep h-3-em-4-ca rboxyl ate, syn isomer prepared in Example 1. 0.335 g of phthalimido acetyl chloride were added, and after 5 minutes at ambient temperature the product was precipitated by the introduction of 10 m] of isopropyl ether, and then recovered byvacuum-filtration, pasting with water and drying. The crude product obtained was agitated for 20 20 minutes at 35-40'C in 2 mi of aqueous formic acid (formic acid: water 2:1), and the product recovered again by adding 10 mi of water, vacuum-filtering, taking up with ether, vacuum-filtering again and then drying. The product thus recovered was further purified by dissolving it in 4 mi of acetone, vacuum-filtering away the insoluble matter and then precipitating with 10 mi of ether. In this way 0.280 g of purified product were obtained.

Analysis: C15H2001ON6S2: 628.16 Calculated: N% 13.77 S% 10.20 Found: 12.510.4 U. V. Spectrum (ethanol) Max. 217 - 218 nm Inf. 230 nm Inf. 238 nm 35 Inf. 260 nm Inf. 300 nm HCl N/10 (ethanol) Max. 217 - 218 nm Inf. 231 nm 40 Max. 239 nm Max. 258 nm Inf. 280 nm N.M.R. Spectrum DIVISO 7.18 p.p.m.: thiazol.

E] = 782 E] = 844 E] = 453 E] = 242 E] = 107.

E' = 773 E' = 429 E' = 352 E1 = 1 286 E] = 230.

Example 3: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2NN-(dimethyl- carbamoyloxyirji:n-o)acetylaminol-ceph-3-em-4-carboxylic acid In 4 mi of methylene chloride were put into suspension under agitation 0. 756 g of diethylamonium 3-acetoxymethyl-7-[2-(2-trityl am i no-th i azol -4-y1)2-hyd roxyi m i no- acetyl am in o]-cep h-3-e m -4-ca rboxyl ate, syn iO isomer, prepared in Example 1.

0.2 mi of pyridine and 0.2 mi of dimethylcarbamoyl chloride were added, and after dissolving, the whole was washed with water and then with water acidified to pH 2.

The organic phase was dried, concentrated to dryness and crushed with ether, after which 0.6 g of crude product was recovered by vacuum filtration. This was agitated for 15 minutes at 4WC with 2 mi of aqueous formic acid (formic acid:water 2: 1), then 5 mi of ethanol were added and the whole was concentrated to dryness to give a residue which was taken up with ethyl acetate. 0.384 g of crude product were recovered and then purified by dissolving in an acetone: methanol mixture (1: 1), adding ether until light floculation occurs, vacuum-filtering away the insoluble matter and precipitating the product from the filtrate with ether.

In this way 0.213 g of purified product were obtained.

Analysis: C181-12008N6S2: 511.52 Calculated: N% 16.4 Found: N% 15.2 so 12 GB 2 101 117 A I.R. Spectrum Nujol (registered Trade Mark) P lactam 1770 cm-1 C=0 1724cm-1 Amide 1667 cm-1 Amide 111532 cm-1 U. V. Spectrum ROH Max. 230 nm Ell = 357 E 18 300 Inf. 251 nm Ell = 261 Max. 300 nm Ell= 87 10 ROH HCl N/10 Max. 256 nm Ell = 270 E 13 800 Inf. 280 nm Ell = 218.

N.M.R. Spectrum DMSO 10.3 p.p.m. = thiazol. 15 Example 4: The,yn isomer of 3acetoxymethyl-7-[2-(2-aminothiazol-4-yl)- 2(phthalimidomethyloxyimino)acetylaminol-ceph-3-em-4-carboxylic acid trifluoroacetate.

Stage A: DiphenyImethyl 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)2-(phthalimido- methyloxyimino)acetylaminol-ceph3-em-4-carboxylate.

At am bient tem perature a mixture containing 0.085 9 of diphenyl methyl 3-acetoxy methyl -7-[2-(2 trityla minoth iazol-4-yi)-2-hyd roxyi m i no-acetyla m inol-cep h-3-em-4- ca rboxyl ate as prepared below, 0.12 g of bromoethyl phthalimide, 0.069 g of potassium carbonate and 0.4 m] of dimethyisulphoxide, was agitated briskly for 15 minutes. 10 m[ of 0.1 N hydrochloric acid were added to form a precipitate, which was vacuum-filtered, washed with water and dried to obtain 0.121 g of crude product. This was dissolved in 1 mi 25 of ethyl acetate and treated with active charcoal, and after eliminating the solvent the residue was crumbled with ether and vacuum-filtered to obtain 0.075 g of purified product.

LR. Spectrum CHC13 NH 3408 cm-1 P lactam 1782 cm-1 phthalimide 1 phthalimide 11 1730 cm 35 ester amide 1665 cm-1 C=C, C=N 1614cm-1 40 aromatic 1599 cm-1 amide 11 1511 cm-1 NH 1493 cm - 45 C-N-OR 1033 cm N.M.R. Spectrum WC13 7.03 p.p.m. aromatics (trityl- and diphenyimethyl) 7.8 p.p.m.: phthalimide 6.9 P.P.M.: C02 CH 3.4 p.p.m.: CH2S Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- (phthalimidomethyloxyim7n-o-)acetylaminol-ceph-3em-4-carboxylic acid trifluoroacetate.

To 0.32 g of the product obtained in Stage A were added 3 mi of trifl uoroacetic acid, and the whole was 60 agitated briskly for 10 minutes at ambient temperature, and then concentrated at WC under reduced pressure. The residue was crumbled in 30 m] of isopropyl ether, vacuum- filtered and rinsed with isopropyl ether to obtain 0.21 g of the expected product.

13 GB 2 101 117 A 13 Preparation: The syn isomer of diphenylmethyl 3-acetoxymethyl-7-[2-(2- tritylaminothiazol-4-yl)-2hydroxyimino-acet71aminol-ceph-3-em-4-carboxylate.

A) The syn isomer of diphenylmethyl 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-(1-methyl- 1 methoxy-ethoxyimino)acetylaminol-ceph-3-em4-carboxylate.

4.15 g of the product obtained in Stage B of the preparation of Example 1 were introduced into 40 CM3 Of 5 methylene chloride and 55 cm' of 0.1 N hydrochloric acid. The whole was agitated for 10 minutes at ambient temperature, and after decanting the organic phase was twice washed with 25 CM3 of water, then dried, vacuum-filtered and rinsed with methylene chloride. Over ten minutes and under agitation, 15 CM3 of 8% diazodiphenyl methane in benzene were added; the whole was agitated for 15 minutes at ambient temperature after which the solvents were evaporated under reduced pressure at 300 to give a residue which 10 was taken up and crumbled in isopropyl ether. After again evaporating the solvent under reduced pressure and taking up with isopropyl ether, the solid was vacuum-filtered, rinsed and dried to obtain 4.41 g of expected product.

N.M.R. Spectrum (CIDC13) 60 MHz c S 11 a = 1.53 p.p.m. b = 2.01 p.p.m. c = 3.26 p.p.m. d = 6.78 p.p.m. e = 7.33 p. p. m.

0 ---, NE 1 C(I 3 (a) N.0 C11 3 (c) N 0 30 C00 f. H2 (c) B) The,yl isomer of diphenylmethyl 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-hydroxyiminoacetylaminol-ceph-3-em-4-carboxylate.

2.775 g of the product obtained above were placed in 14 cm' of acetone and 4.5 CM3 of N hydrochloric acid.

The whole was agitated for two hours at ambient temperature, after which the acetone was driven off under reduced pressure. 20 CM3 of ethyl acetate were added, and after agitating and decanting, the organic phase 45 was washed four times with 10 CM3 of slightly salted water. The wash waters were extracted with 5 CM3 Of ethyl acetate, and the organic fractions were combined and dried, vacuum- filtered, rinsed with ethyl acetate and then evaporated under reduced pressure to drive off the solvent. The residue formed was taken up with ether and crystallised, and the drystals were crumbled, vacuum-filtered and rinsed with ether, to obtain, after drying, 1.88 g of desired product. Rf = 0.5 (eluant = ether with 20% acetone).

N.M.R. Spectrum (C13C13) 60 MHz 6.88 p.p.m. proton of the thiazol cycle 7. 33 p.p.m. proton of the phenyl nuclei.

Example 5: The syn isomer of sodium [3-acetoxymethyl-7-2-(2-aminothiazol4-yl)2-(phthalimido- methyloxyamino) acetyliminol-ceph-3-em-4-carboxylate.

0.21 9 of the product obtained in Example 4 was dissolved in 0.4 mi of methanol, and with agitation, 0.6 mi of a molar solution of sodium acetate in methanol were slowly added to the solution. 2 m] of ethanol were 60 then slowly added, and the resultant precipitate was vacuum-filtered and rinsed with ethanol then ether to give 0.127 g of the desired product.

14 GB 2 101 117 A U. V. Spectrum HCl N/10 (ethanol) Max. 217 nm p 46,500 Inf. 237 nm F 20,000 Max. 252 nm p 16,300 Inf. 301 nm F 7,400 5 Infl 320 nm p 5,850.

N.M.R. Spectrum DIVISO 8.05 p.p.m. aromatic 10 2 p.p.m. 0Ac I.R. Spectrum in nujol C=0 1776 - 1764 -1724 CM-1 Amide 1689 cm-1 14 C=C,C=N 1659 cm-1 15 Aromatic 1611 cm-1 Amide 11 1545cm-1 C07 1525 cm-1 20 1511 cm-1 Example 6: The yn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(a.-carboxybenzyloxy- imino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate.

StageA: Thesyn isomer of diethylammonium 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-((X-tbutyoxycarbonylbenzyloxyimino)acetylaminolceph-3-em-4-carboxylate.

At ambient temperature, a mixture of 1.37 g of 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yi)-2hydroxyimino-acetylaminol-ceph-3-em-4-carboxylic acid, obtained in the course of Stage C of the previous preparation of Example 1, and 20 m] of methylene chloride was agitated. 20 m[ of water were added, and 2.8 mi of triethylamine slowly added. A solution of 4.8 g of t-butyl a-bromo phenyl acetate in 5 mi of methylene 30 chloride was introduced to the emulsion obtained, and the whole was agitated for 23 hours at ambient temperature before acidifying to pH 1 by the addition of 10 mi of 2N hydrochloric acid. After extracting with chloroform, washing the extracts with water, drying and distilling to dryness under reduced pressure at a maximum temperature of WC, 6.1 g of brown oil were obtained. This was dissolved in 5 m[ of ethyl acetate, and 0.25 m[ of diethylamine followed by 50 m] of isopropyl ether were added to the solution. The solid obtained was separated by vacuum-filtration to give 1.11 g of impure product and the mother liquors gave a second portion of 0.1 g of product identical to the first. The two portions of impure product were purified by dissolving in 5 m[ of ethyl acetate, filtering the solution and evaporating the solvent to dryness to give a residue which was taken up with ether recovered again by vacuum- filtration, rinsed with ether and dried at ambient temperature under reduced pressure. 0.96 g of the desired product were obtained, which were used 40 in the following stage without further treatment.

I.R. Spectrum ClHIC13 NH + associate 3372 cm-1 P lactam 1781 cm-1 45 AcO 1738 shoulder C ester 1729 cm-1 11 0 50 amide CW- amide 11 C=N=OR 1681 cm-1 1602 cm-1 1526 cm-1 1493 cm-1 1063 cm - 1 U. V. Spectrum EtO H Inf. 230 nm E 304 Inf. 260 nm E 189 Infl 303 nm E 60 c 9,700 60 Max. 447 nm E] = 8 Inf. 515 nm E] = 3 COH HCl N/10 Max. 267 nrn E] = 160 c 15,000 65 GB 2 101 117 A 15 N.M.R. Spectrum 6.8 p.p.m. thiazol.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(a- carboxybenzyloxyimino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate.

At ambient temperature 0.882 g of the product of Stage A above and 8.9 mi of trif luoroacetic acid were agitated together for 10 minutes. After dissolution the solution was concentrated under reduced pressure at a maximum temperature of WC to a volume of about 3 mi, which volume, after cooling, was taken up with 30 mi of isopropyl ether and agitated therein at ambient temperature for 10 minutes. After vacuum-filtration, rinsing with isopropylether and drying under reduced pressure at 20-250,0. 512 g of impure product were 10 obtained. This was taken up with 2 mi of acetone containing 1% water and agitated therein for 5 minutes at ambient temperature, before slowly diluting under agitation with 20 mi of diethyl ether. After 10 minutes the product was separated by vacuum-filtration, rinsed with diethyl ether and dried at ambient temperature under reduced pressure to give 0.443 g of purified product.

Example 7: The syn isomer of sodium 3-acetoxymethyl-7-[2-(2-aminothiazol4-yl)-2-(acarboxybenzyloxyimino)acetylaminolceph-3-em-4-carboxylate.

At ambient temperature, 0.421 g of the product of Example 6 were dissolved in 2.1 mi of a 1 M methanolic sodium acetate solution and 2.1 mi of methanol. The formed solution was treated with 40 mg of active charcoal and vacuum-filtered on silica, and the filtrate was concentrated under reduced pressure at a maximum temperature of 30'Cto a volume of 1 mi. To this was added 10 mi of 100% ethanol, and the resultant precipitate was separated by vacuum-filtration, rinsed with ethanol then ether and dried under reduced pressure at ambient temperature to give 0. 275 g of the desired product.

Analysis: C231-11909N5S2Na2: 619.5 25 Calculated: C% 44.59 H% 3.09 N% 11.3 S% 10.35 Found: 44.8 3.5 11.3 10.2 U. V. Spectrum EtOH/HCI N/10 Max. 260 nm E' = 266 E 16,500 30 Inf. 276 nm EI = 1 222 F_ 13,700 Inf. 395 rim E] = 8 I.R. Spectrum Nujol P lactam 176cm-1 35 Amide 11 1532 cm-1 C=N-OR 1027 cm-1 Example 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- (2-oxotetrahydrofuran-3yloxyimino)aceiy-la-minolceph-3-em-4-carboxylic acid.

Stage A: Mixture of the A2 and A3 isomers of the syn isomer of diphenylmethyl 3-acetoxymethyl-7-[2-(2 tritylaminothiazol-4-yl)-2-(2-oxotetrahydrofuran-3yloxyimino)acetylaminolce ph-3-em-4-carboxylate.

At ambient temperature, 1.7 g of diphenyimethyl 3-acetoxymethyi-7-[2-(2tritylaminothiazol-4-yi)-2- hyd roxyi m in o-acetyl am i nolceph-3-e m-4-ca rboxyl ate, obtained in the preparation of Example 4, was dissolved in 8.5 mi of dimethylformamide. Under agitation, 0.69 g of anhydrous potassium carbonate then 45 3.2 mi a-bromo-butyrolactone were added, and the resultant suspension was agitated for 45 minutes at ambient temperature and under an inert atmosphere, after which 20 mi of water, 12 mi of N hydrochloric acid and 30 mi of ethyl acetate were added. The organic phase was separated by decanting and washed with water containing sodium chloride until a neutral pH was achieved. After drying and evaporating the solvent to dryness under reduced pressure at a temperature lower than 35', 6.6 g of a clear brown oil were obtained, 50 which was taken up with 20 mi of isopropyl ether and agitated therein for 5 minutes at 40'C. The isopropyl ether phase was separated by decanting, and the operation described above was repeated to obtain a dry extract of about 2.5 g, which was treated in the same way with diethyl ether. The residue was taken up with mi of diethyl,ether and agitated therein at ambient temperature for 15 minutes. The insoluble matter was then separated by vacuum-filtration, rinsed with diethyl ether and dried under reduced pressure to give 1.513 g of an amorphous yellow product, which was used in the following stage without further treatment.

LR. Spectrum NH P lactam 3395 cm-1 1785 cm-1 lactone 0Ac + ester 1744cm-1 Amide 1691 cm-1 16 GB 2 101 117 A 16 C=C C=N 1655 cm-1 Amide H 1599 cm-1 Aromatic 1588 cm-1 5 1515 cm 1493 cm-1 10 N.M.R. Spectrum CD03 6.80 p.p.m. thiazol Stage 8: The syn isomer of diphenylmethyl 1-oxo-3-acetoxymethyl-7-[2- 2tritylaminothiazol-4-yl)2-(2oxotetrahydrofuran-3-yloxy-imino)acetylaminolceph-3-em-4-carboxylate. 15 Under an inert atmosphere and under agitation, 1.525 g of a mixture of the A2 and A3 isomers obtained in the previous stage were dissolved in 8 mi of methylene chloride. The solution was cooled to W- +WC with a bath of iced water, and at that temperature and over a period of 20 minutes 0. 420 g of metachloroperbenzoic acid in 8 mi of methylene chloride were added. The whole was agitated for one hour at the same temperature and concentrated under reduced pressure without heating to give a residue, which was taken up with 20 mi 20 of ethyl acetate and washed therein with a solution of sodium bicarbonate and then with a solution of sodium chloride until neutrality was reached. The organic phase was then dried and distilled to dryness under reduced pressure at a temperature lower than 3WC to obtain a second residue, which was taken up with 10 mi of diethyl ether. The insoluble matter was separated by vacuum-filtration at ambient temperature and rinsed with diethyl ether to give 1.390 g of yellow product.

LR. Spectrum (CH03) NH 3386 cm-1 C=0 lactone + P lactam 1797 cm-1 30 0Ac 1737 cm - 1 conjugated ester amide 1691 cm-1 C=C C=N 35 aromatic 1634 1599 - 1587 - 1523 - 1595 cm-1 Amide 11 S=0 possible 40 1044 - 1035 - 1023 cm-1 + oxime ether U. V. Spectrum (EtOH) Inf. 257 nm Inf. 305 nm (ROH HCl NI,,) Max. 265 nm Inf. 300 nm N.M.R. Spectrum (CDC]3) 2 p.p.m.: 0Ac 6.73 and 6.77 p.p.m.: thiazol Ell = =196 Ell = = 53 E5,000 E] = 200 19,000 Ell == 85 8,100 6.7 p.p.m.: -COOCH 55 7.3 p.p.m. aromatics.

Stage C: The syn isomer of diphenylmethyl3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2(2oxotetrahydrjf-u-ran-3-yloxy-imino)acetylaminolceph-3em-4-carboxylate.

1.328 g of the product obtained in the preceding stage were dissolved in 6.6 mi of dimethyl formamide.

The solution was cooled to -20'C, and under an inert atmosphere 0.6 mi of phosphorus trichloride were added over 30 seconds. The whole was agitated for 5 minutes at -20'C, and then 30 m] of ethyl acetate, 30 m] of a saturated solution of sodium bicarbonate and 15 9 of ice were introduced. After extracting and decanting, the product was re-extracted with 20 m[ of ethyl acetate and washed therein with a solution of 65 17 GB 2 101 117 A 17 sodium chloride until neutrality was reached. After drying and distilling to dryness under reduced pressure and at a temperature lower than 4WC, a residue was obtained which was taken up with 10 mi of diethyl ether, recovered by vacuum-filtration, rinsed and then dried under reduced pressure and at ambient temperature to obtain 1.039 g of product, which was purified on silica by eluting with methylene chloride containing 10% ether. After distilling to dryness and taking up with ether, vacuumfiltering at ambient temperature and 5 drying, 0.751 g of product were obtained.

N.M.R. Spectrum CDC13 2 p.p.m.: 0Ac 6.8 p.p.m.: thiazol (P 6.9 p.p.m.: COOCH 7.3 p.p.m.: aromatics.

Stage D: 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2-oxo tetrahydrofuran3-yloxy-imino)acetylaminolceph-3-em-4-carboxylic acid At 2W-25'C and for 10 minutes, 7.2 mi of trif luoroacetic acid and 0.720 g of diphenyimethyl 3-acetoxymethyi-7-[2-(2-tritylaminothiazol-4-yl)-2-(2-oxotetrahydrofuran3-y loxy-imino)acetylamino]ceph-3- 20 em-4-carboxylate, syn isomer, obtained in the previous stage, were agitated together. The whole was concentrated undir -reduced pressure at a temperature lower than WC to a volume of about 2 mi, which was taken up, while cooling with a water and ice bath, with 30 mi of isopropyl ether. After agitating for 10 minutes at ambient temperature and then vacuum-filtering, 0.483 g of product were obtained. 0.480 g of the product were taken up with 0.5 mi of anisole and 4.8 mi of trifluoroacetic acid, and the whole was agitated for 5 minutes at ambient temperature and then concentrated to a volume of about 1 mi, which was taken up with 20 mi of isopropyl ether. After vacuum-filtering 0.472 g of product were obtained, and to 0.458 g of this were added 1.8 mi of formic acid containing 50% water. The whole was heated to WC for 10 minutes in an inert atmosphere, and whilst still hot was filtered to separate a light insoluble matter. The filtrate was then placed in a water bath not exceeding WC and after cooling was concentrated to dryness under reduced pressure to 30 give a residue which was taken up with 3 mi of water. After vacuum-filtering, rinsing with a very little water and then with ether, 0.283 g of product were obtained and used without further treatment in the following example.

Example 9: The syn isomer of sodium 3-acetoxymethyl-7-[2-(2-aminothiazol4-yl)-2-(2-oxotetrahydrofuran3-yloxy-imino)acetylaminolceph-3-em-4-carboxylate.

0.283 9 of 3-acetoxymethyi-7-[2-(2-aminothiazol-4-yi)-2-(2oxotetrahydrofuran-3-yioxy- imino)acetylaminolceph-3-em-4-carboxylic acid obtained in Example 8 were dissolved in 1 mi of 1 M methanolic sodium acetate solution and 1 mI of methanol. The solution was treated with 30 g of active charcoal, filtered on silica and rinsed 3 times with methanol, and the filtrate obtained was concentrated under reduced pressure to a volume of 1 mf at a temperature not exceeding WC. The 1 mi volume was diluted with 10 mi of ethanol at 100', and after vacuum-filtering, rinsing with ethanol at 100' and then with ether, 0.165 9 of product were obtained.

Analysis: C191-11809N5S2Na: 547.5 45 Calculated: C% 41.68 H % 3.31 N% 12.79 S% 11.72 Na% 4.1 Found: C% 43.8 H% 3.5 N% 11.8 S% 10.7 Na%4.2 U. V. Spectrum ROHMC1 N/10 Inf. 220 nm E] = 255 Max. 260 nm E] = 302 P lactam F 16,500 LR. Spectrum Nujol 1765 cm-1 k lactone Amide C=CC=N coo- Amide 11 1673 cm-1 1611 cm-1 1535 cm-1 18 GB 2 101 117 A N.M.R. Spectrum DIVISO 1. 98 p. p. m.: 0Ac 6.78 p.p.m.: thiazol 4.33 p.p. m.: COOCH2 18 Example 10: The syn isomer of the disodic salt of 3-acetoxymethyl-7-[2-(2aminothiazol-4-yl)-2-(1-carboxy-3hydroxypropoxy-imino)acetylaminolceph-3em-4-carboxylic acid.

0.257 g of sodium salt obtained in Example 9 were agitated with 2.7 m] of a 0.0865 M aqueous solution of sodium carbonate for 4 hours at ambient temperature. After leaving for a further 12 hours, the whole was 10 distilled to dryness under reduced pressure at a maximum temperature of WC to give a residue which was taken up with 2 mi of methanol. The resulting solution was filtered to eliminate the brown insoluble matter and distilled to dryness to give a second residue, which was taken up with 2.5 m[ of ethanol. After recovering the insoluble matter by filtration and rinsing with ethanol then ether, 0.226 g of the desired product were obtained.

Analysis: C19H19010Na2S2Ns 587.5 Calculated: C% 38.84 H% 3.26 N% 11.92 S% 10.91 Na% 7.82 Found: C% 39.8 H% 3.7 N% 10.7 S% 10.1 Na% 7.6 20 U. V. Sp ectrum EtO H/H C 1 NI, 0 Infl 224 nm Ell = 198 Max. 262 nm Ell = 225 E l 5,000 N.M.R. Spectrum D20 25 7.03 p.p.m. thiazol.

I.R. Spectrum lactam 1763 cm-1 amide 1667 cm-1 30 coo- 1575 cm-1 Example 11: The syn isomer of 3-acetoxym ethyl- 7-[2- (2-aminothiazol-4yl)-2-(2-oxo tetrahydrofuran-3-yloxy imino)acetylamino]ceph-3em-4-carboxylic acid.

Stage A: Diethylammonium 3acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)2(2-oxotetrahydrofuran-3- 35 yloxy-imino)acetylamino]ceph-3-em-4-carboxylate, Under agitation and under an inert atmosphere, 1.367 g of 3-acetoxymethyi7-[2-(2-tritylaminothiazol-4yi)-2-hydroxymimino-acetylaminolceph-3-em-4-carboxylic acid, obtained as an intermediate in Stage C of the preparation of Example 1, were dissolved in 20 mI of methylene chloride. 20 mi of water, 2.8 mi of triethylamine and then 1 mi of a-bromo -butyrolactone were added, and the whole was agitated for 17 hours 40 at ambient temperature before adding another 1 mi of a-bromo - butyrolactone followed three hours later by 1.4 m] of triethylamine and, after a further 5 hours' agitation, by yet another 1.5 m[ of a-bromo -butyrolactone and 1.4 m[ of triethylamine. The agitation was then continued for 16 hours, after which 20 m[ of 2N hydrochloric acid were added for acidification. The organic phase was decanted, washed with water until neutral, dried and distilled to dryness under reduced pressure to give a residue which was taken up with 10 45 m] of ethyl acetate. The insoluble matter was separated by vacuum- filtration and rinsed, afterwhich the filtrate (and rinsings) was dried and distilled to dryness. The resultant residue was taken up in 10 mi of ethyl acetate, to which was slowly added 0.2 mi of diethylamine, and the diethylamine salt of the starting product was separated by vacuum-filtration at ambient temperature and rinsed with ethyl acetate then diethyl ether.

The filtrate and rinsings were distilled to dryness and the formed residue was taken up with 2 mI of ethyl 50 acetate, to which was then added 20 mi of isopropyl ether. Finally, the precipitate obtained was recovered by vacuum-filtration to give 0.500 g of the desired product.

U. V. Spectrum EtOH - HCl N/10 Max. 265 nm E] = 179 E 15,000 55 N.M.R. Spectrum (CDC]3) 6.76 p.p.m. thiazol Stage 8: The.yn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2oxotetrahydrofuran3-yloxy- 60 imino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate.

0.5 g of diethylamine salt obtained in Stage A were introduced into 5 CM3 of trifluoroacetic acid, and the whole was agitated for 15 minutes at ambient temperature before distilling under reduced pressure to a volume of about 2 CM3. 20 CM3 of isopropyl ether were then added in one portion, and after agitating for 15 minutes at ambient temperature, separating the product by vacuum- filtration, rinsing 5 times with 2 CM3 Of 65 19 GB 2 101 117 A 19 isopropyl ether and then drying under reduced pressure, the desired product was obtained.

Stage C: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-oxotetrahydrofuran3-yloxy- imino)acetylaminolceph-3-em-4-carboxylic acid.

The trifluoroacetate obtained above was placed in a mixture consisting of 2 CM3 of a 1:1 methylene 5 chloride/methanol solution and 0.5 em 3 of a 1 M ethanolic solution of pyridine, and pasted therein for 15 minutes at ambient temperature. Over a period of two minutes, the whole was diluted with 8 CM3 Of sulphuric ether containing 2% ethanol, and then agitated. The desired product was recovered by vacuum-filtering, ringsing with diethyl ether and drying under reduced pressure.

N% 19.2 S% 6.3 17.8 5.8.

Example 12: The syn isomer of sodium 3acetoxymethyl-7-[2-(2-aminothiazol4-yl)-2-tetrazol-5-ylmethoxyimino)acetylaminolceph-3-em-4-carboxylate. Stage A: The syn isomer of 2-(2tritylaminothiazol-4-yl)-2ftetrbzol-5-ylmethoxyimino)acetic acid.

2.43 9 of 2-(2--tritylaminothiazol-4-yi)-2-cyanomethoxyimino-acetic acid, the preparation of which is described below, were agitated in 12 mi of dimethylformamide. 1.5 g of sodium azide and 1.5 g of ammonium chloride were added and the whole was heated for 5 hours at 750C and then left to cool at ambient temperature before adding 120 ml of distilled water, 40 mi of ethyl acetate and 30 mi of formic acid. The precipitate thus formed was separated by vacuum-filtration, rinsed with water, ethyl acetate then diethyl ether and dried to give 1.275 g of the desired product. 0.127 g of product were recovered from the filtrate.

Analysis: C26H2103N7S = 511.55 Calculated: C% 61.0 H% 4.1 Found: 61.1 4.6 U. V. Spectrum (ROH) (+ DMSO) Inf. 259 nm Inf. 265 nm Inf. 271 rim Inf. 294 nm F_ 11,000 F_ 6,600 ROH / HCl N11, Inf. 270 nm Max. 275 nm E 13,700 N.M.R. Spectrum DMSO 7.29 p.p.m.: trityl 6.88 p.p.m.: thiazol.

I.R. Spectrum Aromatic 1608 em-' 40 NH 1624 em-' C=N N=N 1580 em-' C021557 em-' 1491 em-' 45 Stage& Thesyn_isomerof t-butyl3-acetoxymethyl-7-[2-(2-tritylaminothiazol- 4-yl)-2-(tetrazol-5-ylmethoxyimino)acetylaminolceph-3-em-4-carboxylate.

To 1.024 g of 2-(2-trityiaminothiazoi-4-yi)-2-(tetrazol-5-ylmethoxyimino)acetic acid prepared in the previous stage were added 0.656 mg of t-butyl 7-ami no-3-acetoxymethyi- ceph-3-em-4-carboxyl ate and 2 m] 50 of pyridine. After the addition of 0.5 9 of dicyclohexylcarbodiimide in 5 mi of methylene chloride, the whole was agitated for an hour at ambient temperature, and then the dicyclohexylurea precipitate thus formed was eliminated by vacuum-filtration. 25 m] of N hydrochloric acid were added to the filtrate, and after agitating for 5 minutes, the organic phase was separated by decanting, washed with 25 ml of water and dried. The organic solvent was then distilled away under reduced pressure to give a residue, which was taken up with 55 isopropyl ether and crumbled therein. The solid matter was recovered by vacuum-filtration and rinsed with isopropyl ether, and after drying 1.778 g of crude product were obtained. This was dissolved in 4 m] of ethyl acetate, treated with active charcoal and filtered on silica. Then, over 5 minutes and under agitation, 20 mi of isopropyl ether were added to the filtrate, and the whole agitated for a further 5 minutes. Finally, the solid was separated byvacuum-filtration, rinsed with isopropyl ether and dried to give 1.412 9 of a white product. 60 M.P. = 1580C.

Analysis: C40H3907N9S2 Calculated:

Found:

C% 58.5 H% 4.8 N% 15.3 S% 7.8 58.5 5.0 14.7 7.6 GB 2 101 117 A U. V. Spectrum (ROH - HCI N/10) Max. 267-268 nm c = 19,400 Inf. 290 nm I.R. Spectrum (CHC13) P lactarn 1787 em-' Esters 1738cm-1 Amide 1686cm-1 1673 em N.M.R. Spectrum (C1DC13) 7.2 p.p.m.: trityl 6.75 p.p.m.: thiazo) 1.5 p.p.m.: t-butyl 2.05 p.p.m.: acyl Stage C: The syn isomer of 3-acetoxymethyl7-[2-(2-aminothiazol-4-yl)-2- (tetrazol-5-ylmethoxyimino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate.

At am bient tern peratu re, 1.226 g of t-butyl 3-acetoxym ethyl -7-[2-(2trityl a m inoth iazol-4-yi)-2-(tetrazol-5yimethoxyimino)acetylaminolceph-3-em-4-carboxylate and 12 m] of trifluoroacetic acid were agitated together for 30 minutes, after which the acid was partially evaporated under reduced pressure and a precipitate thrown down by the addition of 120 m] of isopropyl ether. The precipitate was separated by vacuum-filtration, rinsed with isopropyl ether and dried to give 1.061 g of a product containing some starting material, which had only partially reacted, as an impurity. However by repeating the whole operation twice, starting with trifluoroacetic acid, 0.879 g of the desired product were obtained.

Stage D.. The syn isomer of sodium 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(tetrazol-5-yl- methoxyimino)acetylaminolceph-3-em-4-carboxylate. The trifluoroacetate obtained in the previous stage was dissolved in 1.8

CM3 of methanol, and under agitation and at ambient temperature, 2.8 em' of a molar solution of methanolic sodium acetate were slowly added, followed over 5 minutes by 18 em 3 of ethanoi. The agitation was continued for 15 minutes whilst cooling in a bath of ice water, and the solid formed was separated by vacuum-filtration, rinsed with ethanoi then diethyl ether and dried to give 0.447 g of the desired sodium salt in the form of crystals.

Analysis: C17H1707N9S2 MW = 523.5 Calculated: C% 37.4 H% 3.0 N% 23.1 S% Found: 37.8 3.2 21.1 N.M.R. Spectrum (deuterochloroform) 6.73 p.p.m. thiazol 7.25 p.p.m. trityl 11.1 The preparation of the yn isomer of 2-(2-tritylaminothiazol-4-yl)-2- cyanomethoxyimino-acetic acid used as 45 starting material in Stage A.

1) The.yn isomer of cyanomethyl2-(2-tritylamino-thiazol-4-yl)-2cyanomethyloxyimino-acetate.

Under an inert atmosphere, 12.9 g of 2-(2-tritylaminothiazo)-4-yi)-2hydroxyimino-acetic acid, syn isomer, 9.12 g of neutral potassium carbonate, 60 em 3 of dry dimethyl formamide and 7.6 em' of chloroacetonitrile were mixed together, and agitated. The mixture then congealed, after which it was left for 65 hours in an enclosed atmosphere before being poured into a mixture of 750 CM3 of water, 130 CM3 of normal hydrochoric acid and 150 CM3 of ethyl acetate. Afteragitation, the insoluble matterwas separated by vacuum-filtration and rinsed with ethyl acetate and then with water. The phases of the filtrate were separated by decanting, and after washing with 100 em 3 of water and re-extracting with 3 x 100 cm3 of ethyl acetate, the organic phase was dried. Insoluble matter was removed by vacuum- filtration, and after rinsing, the filtrate was concentrated to dryness to give a residue, which was chromatographed on a column of silica, eluting with ether. The ether was then driven off, and 8.69 g of the desired product were obtained in the form of oil.

N.M.R. Spectrum CIDC13 60 MHz Thiazol proton 6.8 p. p. m.

Trityl proton:7.37 p. p. m.

2) The syn isomer 2-(2-tritylaminothiazol-4-yl)2-cyanomethoxyimino-acetic acid 8.69 g of the product obtained in 1) were placed in 52 em 3 of dioxan, and the whole was cooled in an ice 65 21 GB 2 101 117 A 21 bath. Over twenty minutes, 17.1 CM3 of a normal solution of sodium hydroxide were added dropwise, after which the whole was left to reheat spontaneously. 10.5 CM3 of 2N hydrochloric acid were then added, and the dioxan and almost all the water were driven off before adding 20 cm' of water and 30 CM3 of diethyl ether and agitating for 15 minutes. The crystals formed were separated by vacuum-filtration, rinsed with water then diethyl ether and dried to give 4.32 g of the desired product, melting at about 1800C with decomposition.

N.M.R. Spectrum CDC13 60 MHz 4.7 p.p.m.: OCH2CN 6.7 p.p.m.: thiazol proton 7.34 p.p.m.: trityl proton.

Example 13: TheLyn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2benzoyloxyimino- acetylaminolceph-3-em-4-carboxylic acid.

StageA: The yn isomer of diethylaminium 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2benzoyloxyimino-acetylaminolceph-3-em-4-carboxylate.

To a solution of 0.683 g of the syn isomer of 3-acetoxymethy]-7-[2-(2tritylaminothiazol-4-yl)-2hydroxyimino-acetylaminolceph-3-em-4-carboxylic acid in 10CM3 of methylene chloride, were added 0.2 cm 3 of pyridine and 0.2 cm' of benzoyl chloride. The whole was agitated for 10 minutes at ambient temperature and then washed in a separating funnel with water acidified to pH 1, and after drying, vacuum-filtering and concentrating to dryness 0.737 g of a crude product were obtained. This was dissolved 20 in 5 cm 3 of ethyl acetate, and 0, 1 CM3 of diethylamine were added to the solution. After 10 minutes the whole was vacuum-filtered to obtain 0.27 9 of the desired diethylamine salt.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2benzoyloxyimino- acetylaminolceph-3-em-4-carboxylic acid.

The 0.27 g of diethylamine salt were agitated for 10 minutes at 450C with 2 CM3 of 50% aqueous formic acid, after which the whole was distilled to dryness and the residue obtained ground with diethyl ether. 0.155 of crude product were separated and dissolved in 0.5 CM3 Of methanol, from which a solid was precipitated by introducing 5 CM3 of diethyl ether. The solid was separated to give 0. 14 g of desired product.

Analysis: C22H1906N5S2 MW = 513.5 Calculated: N% 13.4 Found: 13.4 Example 14.. The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2-(2aminoethoxyimino)acetylaminolceph-3-em-4-carboxylic acid bis trifluoroacetate. Stage A: The syn isomer of ethyl 2-(2tritylaminothiazol-4-yl)-2-(2-tritylaminoethoxyimino)-acetate. Under an inert atmosphere 12.2 9 of the syn isomer of ethyl 2-(2tritylaminothiazol-4-yi)-2-(2- iodoethoxyimino) acetate were introduced into 80 cm' of anhydrous dimethyl formamide and 12.4 g of triethylamine. The whole was heated to 1 00'C for 5 hours after which 6.2 g of tritylamine were added. The temperature was maintained for a further 7 hours at 1 00'C, before being allowed to return to ambient temperature, at which stage 1600 cm' of distilled water were introduced. The product was extracted with 6 x 250 CM3 of benzene and after washing with water then with a saturated solution of sodium bicarbonate then with a saturated solution of sodium chloride and drying, 23.5 g of a resin were obtained. This was chromatographed on silica, eluting with a benzene ether (95:5) mixture, and the principal was again chromatographed over silica, this time eluting with pure methylene chloride. 3.6 g of pure product were obtained.

N.M.R. Spectrum (CIDC13) Thiazol proton = 6.46 p.p.m. CHAH triplet centred on 2.45 p.p.m. Q = 5 Hz) Stage 8: The syn isomer of 2-(2-tritylaminothiazol-4-yl)-2-(2- tritylaminoethoxyimino) acetic acid.

Whilst under nitrogen, 2 g of the ethyl ester prepared in Stage A were introduced into 10CM3 of dioxan and 55 66 CM3 of absolute ethanol, and 3 CM3 of normal sodium hydroxide solution were added to the formed mixture. After leaving for 65 hours, the precipitate formed was separated by vacuum-filtration and washed three times with 3.5 CM3 of dioxan ethanol (1:6.6) mixture. A first yield of 1.445 g of sodium salt was obtained. The above process, which is one of saponification, was repeated under identical conditions in respect of the mother liquors remaining, which gave a second yield of 0. 440 g of sodium salt.

The 1.445 9 of the first yield was poured into 30 CM3 of water and 30 cm3 of chloroform and, under vigorous agitation, normal hydrochloric acid was added until pH 2 was obtained (about 1.9 CM3). The organic phase was decanted and washed until neutral with 4 X 10 CM3 of water, each fraction of washing water being subsequently re-extracted with 3 cm' of chloroform. The total chloroformic phase was then dried and evaporated to dryness to give a white powder, which was pasted twice with 2 CM3 of dichloroethane and 22 GB 2 101 117 A then twice with 2 CM3 of isopropyl ether. After drying under reduced pressure until a constant weight, 1.202 g of product were obtained. M.p. = 176'C with decomposition.

The second yield of 0.440 g of sodium salt was treated in the same way to give 0.325 g of desired product. M.p. = 176'C with decomposition.

Thus 1.527 g of product were obtained in total.

N.M.R. Spectrum (CDC13) 6.25 p.p.m. proton in 5 of the thiazol 2.95 p.p.m. CH2-N 22 Stage C: The syn isomer diphenylmethyl3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-(2- tritylaminoethoxyimino) acetylaminolceph-3-em-4-carboxylate.

Under nitrogen, 0.286 g of the acid prepared in Stage B were introduced into 2 cm 3 of methylene chloride.

The suspension thus formed was agitated and to it were added dropwise 0.4 cm 3 of a solution prepared extemporaneously by mixing 1.4 cm3 of triethylamine and a quantity of methyl chloride sufficient to obtain 15 cm 3 of solution. The whole was cooled in a bath of acetone and dry ice to -20'C and leftfor 5 minutes to balance the formed solution, after which whilst under agitation 0.4 Crn 3 of a solution prepared extemporaneously by mixing 1.25 cm' of pivaloyl chloride with a sufficient quantity of methylene chloride to obtain 10CM3 of solution, were added dropwise. The bath was subsequently allowed to warm to - 1 O'C, and was left at this temperature for 30 minutes before being allowed to warm further to 10'C in 10 minutes. 0.176 9 20 of cliphenyImethyl 7-amino-cephalosporanate were then immediately alcohol, and the whole was allowed to return to ambient temperature. After leaving for an hour and 20 minutes, a further 17.6 mg of the diphenyl methyl ester were added, and the whole was then agitated for 30 minutes at ambient temperature, placed for hours in a refrigerator, allowed to return to ambient temperature and subsequently evaporated to dryness under reduced pressure. Finally, the residue was filtered on silica, eluting with a benzene ethyl acetate (8:2) mixture, to give 0.208 g of the desired product.

N.M.R. Spectrum (CDC13) 1.99 p.p.m. = O-C-CH3 0 4.38 p.p.m. = N-O-CH2 6.71 p.p.m. = proton in 5 of thiazol 6.88 p.p.m. = C02CH - 02 Stage D: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- (2min aminoethoxyi7 ' o)acetylaminolceph-3-em-4carboxylicaciclbistrifluoroacetate.

186 mg of diphenyimethyl 3-acetoxymethy-7-[2-(2-tritylaminothiazol-4-yl)2-(2-tritylaminoethoxyimino) acetylaminolceph-3-em-4-carboxylate prepared in Stage C was introduced into 1.8 CM3 of pure trifluoroace- tic acid. The yellow solution thus obtained was agitated for 3 minutes at ambient temperature and then, in an 40 inert atmosphere and in an iced water bath, 18 cm 3 of isopropyl ether were rapidly added. The whole was agitated for 10 minutes, after which the solid formed was separated by vacuum-filtration, rinsed with isopropyl ether then diethyl ether and dried to give 100 mg of a white powder. M.P. = about 21 O'C (decomposition).

N.M.R. Spectrum DMSO 2.03 p.p.m. = O-C-CH3 1 0 3.17p.p.m.==N-O-CH2 6.85 p.p.m. = proton in 5Uf thiazol.

Thepreparation of the syn isomerof ethyl2-(2-tritylaminothiazol-4-yl)-2(2-iodoethoxyimino) acetate used as starting material in Stage A.

a) The syn isomer of ethyl2-(2-tritylaminothiazol-4-YI)-2-(2bromoethoxyimino) acetic acid. 55 In an inert atmosphere, a mixture of 4.94 g of ethyl 2-(2- tritylaminothiazol-4-yl)-2-hydroxyimino acetate hydrochloride syn isomer, was introduced into 10CM3 of dimethyl formamide, and at ambient temperature and over 3 minutes, 4.14 g of potassium carbonate were added. The whole was agitated for 20 minutes at 2WC after which 8.65 CM3 of 1,2-dibromoethane were added. After agitation for a further 30 hours the whole was poured into a medium containing 100 CM3 of distilled water and 20 CM3 of methylene chloride. The 60 aqueous and organic phases were then separated by decanting, and after re- extracting with methylene chloride, washing with distilled water and re-extracting again, the combined organic phases were vacuum-filtered, with rinsing, and distilled to dryness. The crude product obtained was chomatographed on silica eluting with benzene containing 5% diethyl ether; and a first fraction was collected which was recrystallised in methanol after dissolution at 50-60'C and vacuum- filtering at 0' - +5'C 1.15 g of cream white 65 23 GB 2 101 117 A 23 product were obtained. M.p. = 1170C.

A homogenous fraction of 1.258 g was subsequently obtained.

N.M.R. Spectrum = p.p.m. (CDC13) triplet = 3.55 J = 7 Hz CHIBr triplet = 4.51 J = 6 Hz N-O-CH2 single 6.55, thiazoi proton cycle.

b) The syn isomer of ethyl 2-(2-tritylaminothiazol-4-yl)-2-(2iodoethoxyimino) acetate.

6 g of ethyl 2-(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxyimino) acetate syn isomer prepared in a) were 10 introduced into 60 em' of methyl ethyl ketone and 2.141 g of sodium iodide. The whole was taken to reflux for one hour and 10 minutes and then evaporated under reduced pressure to give a residue, which was taken up with 120 CM3 of methylene chloride. After washing five times with 40 CM3 of water, each washing being re-extracted with 2 cm3 of methylene chloride, the combined organic phases were dried and evaporated to dryness to give a resin, which was mixed with diethyl ether. After drying under reduced pressure, 6.22 g of 15 product were obtained. M.p. = 1 WC.

N.M.R. Spectrum = (CDC13) CH21 = triplet centred at 3.31 p.p.m. Q - 7 Hz) Proton at 5 of thiazol: 6.53 p.pim.

Example 15.. The.yn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2-(2-aminoethoxyimino) acetylaminolceph-3-em-4-carboxylic acid.

220 mg of the bis trifl uoroacetate prepared i n Exam pie 14 were introduced i nto a tu be and 1.6 em' of a molar solution of sodium acetate in methanol were added. The whole was agitated until complete dissolution was obtained, and then the sides of the tube were rinsed with 0.66 cm3 of methanol before the addition of 18.6 em 3 of absolute ethanol. Precipitation then occurred and at that time an equivalent mixture obtained in the same way but starting with 100 mg of bis trifluoroacetate was added. After 1 hour 50 minutes, the combined mixtures were vacuum-filtered to separate the insoluble matter, which was washed with ethanol then with diethyl ether and dried until constant weight under reduced pressure to give 181 mg 30 of a white powder melting at 27WC with decomposition. Rf = 0.12 (ethyl acetate ethanol water - 60:25:15) The white power was purified as follows:

mg of the powder were introduced into 1 CM3 Of distilled water. After agitating for 5 minutes, pyridine was slowly added until a pH of 7.0 to 7.2 was obtained. After a further 15 minutes of agitation, the whole was vacuum-filtered, rinsed with 0.5 em' of water and 40 em 3 of acetone were added to the filtrate, which was 35 then agitated for 5 minutes, left to rest for 20 minutes and vacuum- filtered to recover the product. After washing three times with acetone and drying 99.5 mg of a purified white powder were obtained.

U. V. Spectrum Max. 261 nm Ell = 348 Example 16: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)- 2-(2-amino-2-thioxoethoxyimino) acetylamino]ceph-3-em-4-carboxylic acid.

0.502 g of sodium 3-acetoxymethyi-7-[2-(2-aminothiazol-4-yi)-2cyanomethoxyimino-acetyilceph-3 -em-4- carboxylate, syn isomer, were placed in 2 em 3 of dimethylformamide and 0. 14 em 3 of triethylamine. After 45 total dissolution, some gaseous hydrogen sulphide was slowly introduced (bubble by bubble) over 15 minutes, and then the whole was left under agitation at ambient temperature for 30 minutes. 20 em' of isopropyl ether were subsequently added and after agitation the isopropyl ether phase was separated, leaving an oil remaining at the bottom of the container. 7 CM3 of ethanol were added to that oil, and after agitation an insoluble substance was formed, which was separated byvacuum- filtration, rinsed with ethanol 50 and dried to isolate 0.388 g of crude product. This was purified by dissolving (0.309 g in 1.5 em 3 of water, adding 30 mg of charcoal, agitating and removing the charcoal by vacuum- filtration whilst rinsing with water. 5 drops of pure formic acid were added to the filtrate, and the precipitated product was rinsed with water and dried to obtain 0.195 g of purified product.

24 GB 2 101 117 A N.M.R. Spectrum: (CH1)2SO 24 (c) 1 5 (b) 10 CIA CC-CH (a) 2 3 (c) CH CO H 0 15 20 (a) single 2.02 p.p.m. (d) single 6.83 p.p.m.

(b) single 3.55 p.p.m. (e) single 7.25 p.p.m.

(c) single 4.75 p.p.m. (f) double centred on 9.73 p.p.m. Q=81-14 25 U. V. Spectrum (in ethanol, N/10 hydrochloric acid) max. 265 nm Ell = 468 c= 24,100 The preparation of the syn isomer of sodium 3-acetoxymethyl-7-[2-(2- aminothiazol-4-yl)-2cyanomethoxyiminoajet-ylaminolceph-3-em-4-carboxylate used in Example 16 as starting material:

A) The syn isomer of t_butyl 3-acetoxym ethyl- 7-[2-(2-tritylaminothiazol4-yl)-2-cyanometh yloxyimino acetylarr7in-olceph-3-em-4-carboxylate.

In an inert atmosphere 1.875 g of the syn isomer of 2-(2tritylaminothiazol-4-yl)-2- cyanom ethyl oxyi m in oacetic acid obtained as in the preparation of Example 12 were mixed with 1.312 g of 35 t-butyl 7-aminocephlosporanate in 12 CM3 of dry methylene chloride. The whole was agitated, and a solution of 960 mg of dicyclohexyl carbodiimide in 12 CM3 of dry methylene chloride added. After agitating and leaving at ambient temperature for one hour 45 minutes, 457 mg of formed clicyclohexylurea were separated by vacuum-filtration. The filtrate was then concentrated to dryness to give a residue, which was chromatorgraphed on silica while eluting with methylene chloride then diethyl ether. The resulting rich fractions were collected, and the diethyl ether therein was driven off, the residue being taken up with ether.

Crystallisation was subsequently initiated, and the whole was left in the refrigerator whilst the product slowly crystallised. The crystals were separated by vacuum-filtration, rinsed with diethyl ether at O'C by pasting and then dried to isolate 776 mg of desired product.

M.P. = 180'C with decomposition.

N.M.R. Spectrum 60MHzWC13 4.9 p. p. m. O-CH2- CN 6.8 p. p. m. proton of the thiazol cycle 7.31 p.p.m. proton of trityl. 50 B) The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- cyanomethyloxyiminoacetylaminolceph-3-en7-4-carboxylic acid trifluoroacetate 779 mg of the product obtained in A) were introduced into 4 CM3 of trifluoroacetic acid, and agitated until dissolution. 17 minutes after the introduction the solution was poured into 40 cm' of isopropyl ether, and 55 after agitation the insoluble matter was separated by vacuum-filtration and dried to isolate 523 mg of the desired product.

c) The syn isomer of sodium 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2cyanomethyloxyimino- acetylat7inolceph-3-em-4-carboxylate.

The 523 mg of trifluoroacetate obtained in B) were dissolved in 2 CM3 of a normal methanlic solution of 60 sodium acetate, and the formed solution was diluted with 6.6 CM3 of ethanol and agitated for 10 minutes.

The insoluble matter was separated by vacuum-filtration, rinsed with ethanol and dried to give 226 mg of the desired sodium salt. M.p. = about 20WC with decomposition.

GB 2 101 117 A 25 N.M.R. Spectrum: CDC13,60 MH, 4.98 p. p. m.: O-CH2- CN 6.86 p. p. m. protoWnof the thiazol cycle.

Analysis: C17H1507N6S2Na Calculated C% 40.64 H% 3.01 Na% 4.57 5 Found: 40.2 3.3 4.5 Example17: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- (2amino-2thioxoethoxyimino)a-cetylaminolceph-3-em-4-carboxylic acid. StageA: 2-(2tritylaminothiazol-4-yl)-2-(2-amino-2-thioxo-ethoxyimino) acetic acid.

2.814 g of 2-(2-trityl am in oth lazo 1-4-yl)-2-(cya no m eth oxyi m in o) acetic acid obtained in the preparation of Example 12 were introduced into 12 cm 3 of a solution containing 1.6 cml of triethylamine diluted with 20 CM3 of dimethylformamide. At ambient temperature gaseous hydrogen sulphide was introduced for 30 minutes, after which the mixture was corked and left for 2 hours 10 minutes under agitation. 100 CM3 of water 1 followed by 14.4 CM3 of normal hydrochloric acid were then added, and the whole was agitated vigorously before removing the insoluble matter formed by vacuum-filtration and rinsing with water. Methylene chloride was added to the filtrate, and the crystals formed were separated by vacuumfiltration, rinsed and dried to isolate 2.47 g of the desired acid. M.P. = 18WC.

N.M.R. Spectrum - (CH3)2SO 6.96 p.p.m.: trityl, 7.33 p.p.m. proton in 5 of thiazol.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4yl)-2-(2-amino-2-thioxo- ethoxymino)ic-etylaminolceph-3-em-4-carboxylic acid.

1.01 g of the acid obtained in Stage A and 0.656 mg of 7aminocephalosporanic acid were introduced into 2 CM3 of pyridine. After dissolution 0.5 g of dicyclohexyl carbodiimide in 5 CM3 of methylene chloride were added, and the whole was agitated for one hour at ambient temperature. The formed precipitate was then removed by vacuum-filtration, rinsing with methylene chloride, and dried to isolate about 0.5 g of dicyclohexylurea. The filtrate was poured into 25 CM3 of normal hydrochloric acid. After agitating for 5 minutes, the organic phase was separated by decanting, washed with 25 CM3 of water, dried and vacuum-filtered and the solvent was driven off from the filtrate to leave a residue, which was taken up and crumbled in isopropyl ether. The insoluble matter was then separated by vacuum-filtration and rinsed to obtain the desired product in impure form. This was dissolved in 4 CM3 of ethyl acetate, and 0. 18 g of active charcoal were added, which were subsequently removed by vacuum-filtration, rinsing with ethyl acetate. 35 The filtrate was agitated, and over 5 minutes 20 cm3 of isopropyl ether were introduced before continuing the agitation for a further 5 minutes. The insoluble matter was recovered by vacuum-filtration, rinsing with isopropyl ether, to obtain after drying the desired purified product.

StageC: The syn isomer of 3-acetoxymethyl-7-[2-(2aminothiazol4-yl)-2-(2amino-2-thioxoethoxyimino)aje-t-ylaminolceph-3-em-4-carboxylic acid.

The product obtained in Stage B was introduced into 12 cm' of trifluoroacetic acid, and the whole was agitated for 30 minutes at ambient temperature. The acid was partially evaporated under reduced pressure and precipitation was initiated with 120 cm.3 of isopropyl ether. The precipitate was crumbled and then recovered byvacuum-filtration, rinsing with isopropyl ether, to obtain after drying, a product identical to that 45 of Example 16.

Example18: The syn isomer of 3-acetoxymethyl-7-[2-(2- aminothiazol-4-yl)2-(2-oxopropoxyimino)acetylaminolceph-3-em4-carboxylic acid trifluoroacetate.

StageA: The syn isomer of 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4yl)-2-(2-oxopropoxyimino)acetylaminolceph-3-em-4-carboxylic acid.

7.6 g of diethylammonium 3-acetoxymethyi-7-[2-(2-tritylaminothiazol-4-yi)2-hydroxyaminoacetylaminolceph-3-em-4-carboxylic acid, syn isomer, prepared in Example 1, was introduced into 114 CM3 of methylene chloride and 114 CM3 of distilled water. Then at 200C and under agitation, 14 cm3 of triethylamine were added. The whole was agitated for 5 minutes, after which 8.45 CM3 of bromoacetone were added at 2W-25'C. After agitation at 25'Cfor 5 hours, the whole was acidified to pH 1-2 at 15'C with 6 cm 3 of hydrochloric acid. The organic phase was separated by decanting, and produetwas extracted from the aqueous phase with 250 CM3 methylene chloride. The organic phases were combined and washed with water, and after re-extracting the product from the washing waters, the total organic phase was dried and distilled to dryness to obtain 8.48 g of a resin. This was dissolved in 8. 5 CM3 of methylene chloride, and precipitation therefrom as then induced bythe addition of 85 CM3 of diethyl ether. After agitation for 2 hours at ambient temperature, the precipitate was separated by vacuum-filtration, rinsed three times with 10CM3 of ether and dried to obtain 6.27 g of the desired product.

26 GB 2 101 117 A 26 Purification by converting to the diethylammonium salt:

The product obtained above was taken up in 62 CM3 of ethyl acetate, and pasted thereon at ambient temperature for 5 minutes. Insoluble matter was removed by vacuum-filtration, rinsing with ethyl acetate, and the filtrate was concentrated to 50 CM3 before the introduction in one portion of 0.6 CM3 of diethylamine.

After agitation, the gum formed was rinsed three times with 5 CM3 of ethyl acetate. The filtrate was diluted with 130 CM3 of isopropyl ether, and the precipitate thus formed was agitated in the mother liquorfor 15 minutes before being separated by vacuum-filtration, rinsing with isopropyl ether. After drying, 3.27 g of the desired salt were obtained.

3.25 g of this salt were dissolved in 25 CM3 of distilled water and 98 CM3 of ethyl acetate, and 50 CM3 of a saturated aqueous solution of sodium chloride were added. One gum was decanted, it being recovered with 10 the aqueous phase. After acidification to pH 1-2 with 2.5 CM3 of concentrated hydrochloric acid, in the presence of 50 CM3 methylene chloride, product was extracted from the aqueous phase with methylene chloride (50 CM3 twice). The organic phase was then washed with water, dried, vacuum-filtered with rinsing and distilled to obtain 1.79 g of a resinous product. This was dissolved in 4 CM3 of methylene chloride, and after dilution with 40 CM3 of diethyl ether, the insoluble matter was recovered by vacuum-filtration, rinsed 15 with diethyl ether and dried to obtain 1.22 g of the desired product.

N.M.R. Spectrum - CDC13 2.03 p. p. m. O-C-CH3 1 20 0 2.13 p.p.m.CH2-C-CH3 0 6.8 p.p.m. proton in 5 of thiazol.

StageB: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2oxo- prop oxyimino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate.

1.22 g of the product obtained in Stage A were introduced into 12 CM3 of trifluoroacetic acid and agitated therein for 15 minutes at 20-22'C. The whole was reduced in vacuo to 6 CM3 and then diluted with 60 CM3 Of isopropyl ether, and the precipitate thus formed was agitated at ambient temperature for 15 minutes before being separated by vacuum-filtration, rinsing five times with 5 CM3 of isopropyl ether, to obtain, after drying under reduced pressure, 0.97 g of crude product.

This was purified by dissolving in 4 CM3 of acetone containing 1 % water and then adding 0.2 9 of active charcoal, which, after agitating for 5 minutes at 20-22'C, was separated by filtration and rinsing five times with 1 CM3 of acetone containing 1% water. At 2W-22'C, 90 CM3 of diethyl ether were added to the filtrate, and after agitation at 20-22'C for one hour the insoluble matter was separated by vacuum-filtration, rinsing with diethyl ether and dried to obtain 0.57 g of the desired product.

Example19: The syn isomer of sodium 3-acetoxymethyl-7-[2-(2-aminothiazol4-yl)-2-(2-oxoprop oxyimino)acetylamino]ceph-3-em-4-carboxylate. 0.57 9 of the trifluoroacetate obtained in Example 18 was dissolved in a mixture of 0.25 CM3 of distilled 45 water, 2.3 CM3 of methanol and 2.3 cm' of a normal methanlic solution of sodium acetate. The light insoluble 45 matter obtained was removed by vacuum-filtration rinsing with 2.3 CM3 of methanol, and the filtrate was concentrated to about 3 CM3 to obtain a coloured solution, which was diluted with 20 CM3 of ethanol. After agitating at 20-22'C for 10 minutes, the insoluble matter was separated by vacuum-filtration, rinsing three times with 3 CM3 of ethanol, and dried to obtain 0.38 g of the desired sodium salt. 50 By concentrating the mother liquors it was possible to obtain by vacuum-filtration a second portion of 0.12 50 g of sodium salt, which was pasted at 20'C for 5 minutes in 0.6 CM3 of methanol. After dilution with 6 CM3 Of ethanol and subsequently rinsing, 0.07 g of sodium salt were obtained. The sodium salt thus obtained (0.45 g) was purified by dissolving at ambient temperature in 2 cm' of distilled water, and slowly and under agitation at 2WC adding 12 CM3 of acetone. The precipitated gums were removed by vacuum-filtration, and the filtrate was brought to dryness to obtain a reside. The residue was 55 treated as just-described, starting with 1.5 cM3 of distilled water and 10CM3 of acetone, and the resulting residue was taken up with 5 CM3 of pure ethanol. The insoluble matter was then vacuum-filtered at 20'C, rinsing three times with 1 cm 3 of ethanol, and dried under reduced pressure to obtain 0.26 g of the desired product.

N.M.R. Spectrum (CH3)SSO 2 p.p.m. O-C-CH3 0 27 GB 2 101 117 A 27 2.13 p. p. m. CH2-C-CH3 0 6.78 p.p.m.: proton at5 position ofthiazol. 5 Example20: The syn isomer of 3-methyl-7-[2-(2-aminothiazol-4-yl)-2-(2aminoethoxyimino)acetylamino]ceph-3-em-4-carboxylic acid bis trifluoroacetate. StageA: The syn isomer of diphenylmethyl3-methyl-7-[2(2-tritylaminothiazol-4-yl)-2-(2-tritylaminoethoxyimino)acetylaminolceph-3-em-4-carboxylate.

0.923 g of the syn isomer of 2-(2-trityiaminothiazol-4-yi)-2-(2tritylamino-ethoxyimino)acetic acid, prepared according to the methoddescribed in Stage B of Example 14, were introduced into and then dissolved in 6.5 cm 3 of methylene chloride and 1.3 cm 3 of a solution of triethylamine, the triethylamine solution being prepared by mixing 1.4 CM3 of triethylamine and a sufficient quantity of methylene chloride to obtain 10 cm3 of solution. After dissolution, the whole was cooled to -20'C, and 1.3 CM3 of a solution of pivaloyl chloride 15 (prepared by mixing 1.25 cm' of pivaloyl and a quantity of methylene chloride sufficient to obtain 10 cm3 of solution) were added. The temperature was then allowed to rise to 1 O'C and maintained there fore 35 minutes, before being allowed to rise further to +1 O'C, at which point 0. 494 g of benzhydryl 7-amino-desacetoxy-cephalosporanic acid were added. The whole was left to attain ambient temperature and was agitated for 1 hour 20 minutes, after which a further 77 mg of benzhydryl 7-amino-desacetoxy- 20 cephalosporanic acid were added. After agitation for one hour and evaporating to dryness under reduced pressure, the residue was chromatographed on silica, efuting with a benzene ethyl acetate mixture (8:2), to obtain 0.504 of the desired product.

N.M.R. Spectrum (CDC13) 2.1 p.p.m. methyl at3 position 6.93p.p.m. proton of di phenyl methyl CH02 StageB: The syn isomer of 3-methyl-7-[2-(2-aminothiazol-4-yl)-2-(2-amino- ethoxyimino)acetylaminolceph-3-em-4-carboxylic acid bis trifluoroacetate.

* 357 mg of the product obtained in Stage A were introduced into 3 CM3 of trifluoroacetic acid, and the solution formed was agitated for 2 hours 30 minutes and then placed in a bath of iced water before quickly adding under agitation 40 c& of a mixture (50:50) of isopropyl ether and petroleum ether (fraction 64-75'C).

The whole was agitated for 10 minutes and the insoluble matter was then vacuum-filtered, rinsed with isopropyl ether then diethyl ether and dried until constant weight to obtain 200 mg of white powder. M.p. 35 about 250'C with decomposition.

N.M.R. Spectrum (CH3)2SO 2.03p.p.m. methyl at3 position 6.88p.p.m. proton at5 position ofthiazoi.

Example21: The syn isomer of 3-acetoxymethyl-7[2-(2-aminothiazol-4-yl)-2(2bromoethoxyimino)acetylaminolceph-3-em-4-carboxylic acid trifluoroacetate. StageA: The syn isomer of t-butyl 3-acetoxymethyl-7-[2(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxyimino)acetylaminolceph3-em-4-carboxylate.

1.185 g of the syn isomer of 2-(2-tritylaminothiazol-4-yi)-2-(2-bromoethoxyimino)acetic acid and 0.725 g of t-butyl 7-amino-cephalosporanate were introduced into 20 cml of methylene chloride. The whole was cooled, and then under an inert atmosphere, 4.75 CM3 of 0.5M dicyclohexylcarbodlimide solution in methylene chloride were slowly introduced. After agitating at 00 - +5'C for one hour 30 minutes dicyclohexylurea crystallises out, which after agitating for a further one hour at 20-25'C was separated by 50 vacuum-filtration at 20'C, rinsing with methylene chloride. 0.293 g of dicyclohexylurea were obtained. The filtrate was brought ot dryness under reduced pressure to obtain 2.09 g of impure product, which was purified by chromatography on silica, eiuting with a mixture of benzene and sulphuric acid (1: 1) to obtain 1.129 g of the desired product.

StageB: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2-(2bromo- ethoxyimino)acetylaminoIceph-3-em-4-carboxylic acid trifluoroacetate.

1.301 g of product obtained from Stage A were introduced into 13 CM3 of trifluoroacetic acid, and the whole was agitated at ambient temperature for 10 minutes, with total dissolution occurring after one minute of agitation in an inert atmosphere. The formed solution was distilled under reduced pressure to drive off 60 trifluoroacetic acid whilst in a water bath at a temperature lower than 35'C, the distillation being continued until a residual volume of about 5 CM3 was obtained. Whilst cooling further with an ice bath and whilst under agitation, 42 CM3 of isopropyl ether were added, and a yellow precipitate was obtained, which after agitation at ambient temperature for a further 30 minutes was separated by vacuum- filtration, rinsed three times with 2.6 CM3 of isopropyl ether and dried under reduced pressure at ambient temperature to obtain 0.893 g of the65 28 GB 2 101 117 A desired product.

28 The preparation of the syn isomer of 2-(2-tritylaminothiazol-4-yl)-2-(2- bromo-ethoxyimino) acetic acid used as starting material in Stage A.

a) The syn isomer of ethyl 2-(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxyimino) acetate.

Under argon, a mixture of 4.94 g of ethyl 2-(2-tritylamino-4-thiazoiyi)-2hydroxyimino-acetate hydrochloride, syn isomer, were introduced into 10CM3 of dimethyl formamide, and at ambient temperature and over 3 minutes, 4.14 g of potassium carbonate were added. The whole was agitated for 20 minutes at 20'C before the addition of 8.65 cm' of 1,2- dibromoethane, and after agitation for a further 30 hours, the whole was poured into a medium containing 100CM3 of distilled water and 20 CM3 of methylene chloride. The organic 10 aqueous phased were separated by decanting before extracting product from the aqueous phase with methylene chloride, washing the combined organic phases with distilled water and re-extracting product from the washings. The entire organic phase was then dried, vacuum-filtered and distilled to dryness to give a crude product, which was chromatographed on silica, eluting with benzene containing 5% diethyl ether. A first fraction was collected, with was reerystallised from methanol after dissolving at 50-60'C and vacuum-filtering at 0' - +5'C. 1.16 9 of cream white product were obtained, m.p. = 117'C, and subsequently a homogenous fraction of 1.258 g was obtained.

b) The syn isomer of2(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxyimino)acetic acid.

Under argon, 2.88 g of ethyl 2-(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxyimino) acetate, syn isomer, 20 obtained from a), were introduced into 4.3 cm' of dioxan. The whole was agitated at ambient temperature in an inert atmosphere until dissolution after which over 3 minutes and at 20-25'C were added 30.6 CM3 of 0.5M ethanolic solution of potassium in ethanol. After agitating at ambient temperature in an inert atmosphere for 24 hours, the potassium salt, which had crystallised out after 2 to 3 hours of reaction, was separated by vacuum-filtration at ambient temperature, rinsing with 1 CM3 of diethyl ether then three times with 2 CM3 Of 25 methylene chloride, to obtain 1.6 g of the desired salt. This was taken up with a mixture of 10 c& of distilled water, 10CM3 of methylene chloride and 1 CM3 of 2N hydrochloric acid, which was then agitated in an ampoule for several minutes before the addition of 10 CM3 of distilled water, 10CM3 of methylene chloride and 1 cm 3 of 2N hydrochloric acid. The methylene chloride phase was washed to neutrality with three times 20 CM3 of distilled water, and the product was re-extracted from the washing waters with 10 CM3 Of methylene chloride. The methylene chloride phases were then combined, dried, vacuum-filtered with rinsing and distilling to dryness to obtain 1.465 g of resin. This was purified by taking up with 15 CM3 Of 1,2-dichloroethane and dissolving therein whilst heating to about 4WC. The temperature was then brought down to 2WC, when crystallisation started and the whole was agitated for 3 hours at 2WC. After vacuum-filtration, rinsing with 0.5 CM3 Of 1 2-dichloroethane and dryness 1.185 g of a white product were 35 obtained.

M.P. = 15WC. Rf. = 0.65 (acetone containing 5% water).

Analysis:

Calculated: C% 58.21 H% 4.13 N% 7.82 S% 5.98 Br% 14.89 40 Found: C% 58.0 H% 4.2 N % 7.8 S% 5.9 Br%15.2 N.M.R. Spectrum WC13 P.P.M.

3.44 (triplet) -CH2-Br J = 7.5 Hz 4.3 (triplet) -N--:-0-CH2- J = 7 Hz 6.55 single: proton nn-position of the thiazole.

Example22: Sodium salt of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2bromoethoxyimino)- acetylaminol-ceph-3-em-4-carboxylic acid, gyn isomer.

0.893 g of the trifluoroacetate obtained in Example 21 and 3.5 CM3 of a molar methanolic solution of sodium acetate were mixed and agitated at ambient temperature under an inert atmosphere until dissolved, then filtered and rinsed first with 1 CM3, then twice with 0.5 cm', of pure anhydrous methanol. 40 CM3 Of absolute ethanol were added. Some cloudiness was observed and then the sodium salt precipitated. After agitation for 2 hours at ambient temperature, the salt formed was vacuum- filtered and rinsed three times with 1 cm 3 of absolute ethanol then three times with 3 CM3 of diethyl ether. The sal was dried at ambient temperature under reduced pressure to obtain 0.545 g of product. Rf. = 0.6 acetone containing 10% water [a120 = D+45' 1.5' (c = 1 % in H20) Analysis:

Calculated: C% 35.80 H% 3.00 N% 12.28 S% 11.24 BR% 14.01 60 Found: C% 36.0 H % 3.1 N% 11.9 S% 11.2 Br% 13.7 N.M.R. Spectrum (CID3)2SO. (P.P.M.) 29 GB 2 101 117 A 29 2.0: -0-C-CH3 11 0 3.62 (triplet): -CH2-Br J = 6Hz 4.33 (triplet): =N-O-CH2- J = 611z 6.75: proton in 5-position of the thiazole.

Example23: The syn isomer of 3-acetoxymethyl-7-(2-(2-aminothiazol-4-yl)[2-(2-[2-aminophenylthiolethoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid. StageA: The syn isomer of 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-(2-[2-aminophenylthiolethoxy-imino)-jc-etylaminol-ceph-3-em-4-carboxylic acid.

Successively 790 mg of 3-acetoxym ethyl -7-[2-(2-tritya 1 m in oth iazol4-yi)-2-(2-bro moethoxy-i m i no)- acetylaminol-ceph-3-em-4-carboxyiic acid, 8 mi of benzene and 0.28 mi of pure triethylamine were introduced into the reaction vessel. Aftertotal dissolution, 0.14 m] of 2amino-thiophenol, 5 mi of distilled water and several mg of methyltricaprylammonium chloride were added. The mixture was agitated briskly for one night, then 2 mi of 1 M hydrochloric acid was added and the mixture was extracted with ethyl acetate, washed with water, dried and concentrated to dryness. The residue was taken up with diethyl ether, and 20 vacuum-filtered to obtain 690 mg of 3-acetoxymethy-7-[2-(2tritylaminothiazol-4-yi)-2-(2-[2-aminophenyi thio]-ethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid.

StageB: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2[2-aminophenyl-thioll- ethoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid.

To the product obtained in Stage A 5.3 mi of 66% aqueous formic acid were added, and the mixture was heated to 550C for 15 minutes, cooled, and vacuum-filtered. The solvents were driven off from the filtrate by distillation under reduced pressure, and the residue was taken up with ethanol then distilled to dryness under reduced pressure. The desired product was obtained and used without further purification in the following example.

The starting material for Stage A of this Example was prepared as follows.

26.85 g of the syn isomer of 2-(2-tritylaminothiazol-4-yi)-2-(2bromoethoxy-imino)-acetic acid prepared in Example 1, 7.25 g of 1-hydroxy 1 H-benzo-triazol, 12 g of dicycloexylcarbodiimide and 350 mi of anhydrous methylene chloride were mixed and agitated for 23 hours at ambient temperature. The dicyclohexylurea thus formed was removed by vacuum-filtration, and the filtrate was washed with water, with a 1 M aqueous solution of sodium bicarbonate, then with water. The filtrate was dried, then concentrated to dryness under reduced pressure. The residue was taken up with 150 mi of ether and 25.4 g of crystals were obtained which were dissolved in 235 mi of anhydrous methylene chloride. To the solution were added 10.56 g of 7-amino-cephalosporanic acid, 200 mi of anhyrous methylene chloride and 10.9 mi of triethylamine, and the mixture was agitated for 65 hours. 350 mi of water and 45 mf of 2N hydrochloric acid were then added and 40 the formed mixture was agitated to effect extraction. The organic phase was washed then dried and concentrated to dryness under reduced pressure. The residue was taken up with 75 mi of ethyl acetate and diluted with 520 mi of diethyl ether, then vacuum-filtered to obtain 24. 75 g of the desired product.

Example24: Sodium saftof3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2[2-aminophenyl-thiol-e thoxy- 45 imino)-acetylaminolceph-3-em-carboxylic acid, syn isomer.

To the product obtained in Example 23 there were added 1.5 m[ of a molar methanolic solution of sodium acetate and 1 mi of methanol. This was mixed, then diluted with 6 mi of ethanol. The precipitated product was vacuum-filtered off, washed with ethyl alcohol, then made into a paste with ether to yield 235 mg of the desired product. 50 Analysis C23H2307N6S3Na pM 614.65 Calculated C% 44.94 H% 3.77 N% 13.67 S% 15.65 Found 44.8 3.7 13.3 14.8 55 N.M.R. Spectrum (CID3)2SO (P.P.M.) 1.99 -0-C-CH3 11 0 60 -4.08 =N-O-CH2- 6.73 proton in 5-position of thiazole.

GB 2 101 117 A Example25: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2([2-amino-1,3,4-thiadiazol-5yll-thio-ethoxy-imii-o)--acetylaminol-ceph-3-em-4-carboxylic acid.

StageA: 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-([2-amino-1,3, 4-thiadiazo l-5-yl-thiol-ethoxy- imino)-acetylaminol-ceph-3em4-carboxylic acid.

a) 1.9 g of 2-amino-5-thiol-1,3,4-thiadiazole were mixed with 15 m[ of a molar solution of lithium methylate 5 in methanol. After total dissolution, the solution as concentrated to dryness under reduced pressure to obtain the lithium derivative of 2-amino-5-thiol-1,3,4-thiadiazole in the form of a resin which was used in that form.

b) 7.91 g of the syn isomer of 3-acetoxymethy]-7-[2-(2-tritylaminothiazol4-yl)-2-(2-bromoethoxy-imino)- acetylaminol-ceph-3-em-4-carboxylic acid prepared in Example 3 were introduced into 30 mi of anhydrous 10 dimethylformamide and 670 mg of lithium iodide. After dissolution the lithium derivative obtained above was added with 15 m[ of dimethylformamide. The mixture was agitated for 5 and a half hours at ambient temperature, then 450 m] of water and 1 m] of formic acid were added. Then the mixture was extracted with ethyl acetate, dried and concentrated to dryness under reduced pressure. The residue was chromatographed on silica eluting with a 70-20-10-1 ethyl acetate-methanol-water- triethylamine mixture, to obtained 2.36 g of 15 the desired product.

StageB: 3-acetoxymethyl-7-[2-(2-aminothiazol4-YI)-2-([2-amino-1,3,4thiadiazo1-5-yl -thiol-ethoxy-imino)- acetylaminol-ceph-3-em-4-carboxylic acid.

843 mg of the tritylated product of Stage A were agitated for 15 minutes at WC with 6.4 mI of 66% 20 aqueous formic acid. The mixture was concentrated to dryness under reduced pressure and the residue was taken up with 5 m[ of ethanol and 50 mi of diethyl ether, then vacuum- filtered to obtain 606 mg of desired crude product.

Example26: Sodium salt of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2([2-amino-1,3,4-thiadjazo1-5-yl thiol-ethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer.

The product obtained in Example 25 was mixed with 1.5 mi of-molar sodium acetate in methanol and 0.5 m] of dimethy[formamide. 5 m[ of ethanol were added and the precipitate thus formed was vacuum-filtered off, taken up with 7 m[ methanol and refluxed, then cooled and vacuum- filtered to remove impurities. The filtrate was concentrated to dryness under reduced pressure, and the residue was broken up in ethanol, vacuum-filtered and dried to obtain 146 mg of the desired product.

Analysis C191-11907NSS4Na PM:622.66 Calculated C% 36.65 H% 3.08 N% 17.99 S% 20.60 Found: 37.0 3.7 17.5 19.8 35 N.M.R. Spectrum (CID3)2SO p.p.rn.

1.99 -0-C-CH3 0 -4.25 =N-O-CH2 6.76 proton in 5-position of thiazole. 45 Example27: 3-acetoxymethyl-7-[2(2-aminothiazol-4-ylj-2-(2-[5-nitro-2pyridinyl-thio]-e thoxy-imino) acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer. StageA: 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-(2[5-nitro2-pyridinyl-t hiolethoxy-iminojacetylaminol-ceph-3-em-4-carboxylic acid.

Into a reaction vessel, 396 mg of 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yi)-2-(2-bromoethoxyimino)-acetylamino]-ceph-3-em-4carboxylic acid prepared in Example 3,4 m] of benzene and 0.14 mi of triethyalmine were successively introduced.

After dissolution 92 mg of 2-thiol-5-nitro-pyridine, 5 mi of distilled water and several mg of methyltricaprylammonium chloride were added, and the mixture was agitated for 72 hours at ambient temperature. Then 1 mi of N hydrochloric acid was added and the whole extracted with ethyl acetate. The organic phase was dried then concentrated to dryness under reduced pressure. The residue was taken up with diethyl ether, and vacuum-filtered to obtain 416 mg of the tritylated product.

Stage 8: 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2-[5-nitro-2pyridinyl-thiol-e thoxy-imino)- 60 acetylaminol-ceph-3-em-4-carboxylic acid.

To the product obtained there were added 3.2 mi of 66% aqueous formic acid and the mixture was heated to WC for 15 minutes. The triphenylcarbinol thus formed was removed by vacuum-filtration, and the filtrate was concentrated to dryness under reduced pressure. The residue was taken up with ethanol and concentrated to dryness again to obtain the desired product.

31 GB 2 101 117 A 31 Example28: Sodium saftof 3-acetoxy-7-[2-(2-aminothiazol-4-yl)-2-(2-[5nitro-2-pyridinyl-thioll-ethoxy imino)-acetylaminol-ceph-3-em-4carboxylic acid, syn isomer.

A little methanol was added to the product obtained in Example 27, followed by 2 m] of a molar methanolic solution of sodium acetate. The traces of insoluble matter were removed by vacuum-filtration, and the filtrate was concentrated to dryness under reduced pressure. The residue was taken up with ethanol, and vacuum-filtered to obtain 153 mg of the desired sodium salt.

Analysis C22H2009N7S3Na PM = 645.63 Calculated C% 40.93 H% 3.12 N% 15.1 S% 14.90 Found: 39.0 3.3 13.2 12.9 10 N.M.R. Spectrum (C133)2SO P.P.M.

2.0: -0-C-CH3 11 0 =3.86 - 4.3 - 4.4: =N-O-CH2- 6.76: proton in 5-position of thiazole. 20 Example 29: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2-(-[3-cyano-6-methyl-2pyridinylthiol-etyoxy-imino)-acetylaminol-ceph-3em-4-carboxylic acid Stage A: 3-acetoxymethyl-7-[2-(2-tritylaminothiazol4-yl)-2-(2-[3-cyano 6-methyl-2-pyridinyl-thiol-ethoxyimino)-acetylaminolceph-3-em-4-carboxyylic acid.

791 mg of 3-acetoxymethyl-7-[2-(2-tritylamino-thiazol-4-yi)-2-(2bromoethoxy-imino)-ac etylamino]-ceph3-em-4-carboxylic acid prepared in Example 23 were mixed with 326 mg of 2-th i of-3-cya n o-6-m ethylpyridine, 10 mi of benzene, 0.42 mi of triethylamine, 10 mi of water and several milligrams of methyltricaprylammonium chloride. The mixture was agitated for 44 hours at ambient temperature, then acidified with 3 mi of N hydrochloric acid and extracted with ethyl acetate. The organic phase was dried and 30 concentrated to dryness under reduced pressure. The residue was taken up with diethyl ether, and vacuum- filtered to obtain 877n mg of a crude product.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-[3-cyano-6-methyl-2-pyridiny l35 thiol-ethoxy-imino)-acetylamino-ceph3-em-4-carboxylic acid.

To the product obtained in Stage A there was added 7 mi of 66% aqueous formic acid and the mixture was heated to WC for 15 minutes with agitation, then cooled and vacuum-filtered. The solvents were driven off from the filtrate by distillation under reduced pressure, and the residue was taken up with ethanol and concentrated to dryness to obtain a crude product which was purified by forming the sodium salt and 40 treating this with formic acid.

Analaysis: C24H2307N7S3 PM = 617.69 Calculated C% 46.67 1-1% 3.75 N% 15.87 S% 15.57 Found: 46.2 3.8 15.2 14.5 N.M.R. Spectrum (DMSO) p.p.m.

2.0: O-C-CH, 0 = 4.2 - 4.3 - 4.4: =N-O-CH2 6.8: proton in 5-position of f-hiazole.

Example 30: Sodium salt of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-[3-cyano-6-methyl-2-pyridiny lthiol-ethoxy-imino)acetylaminol-ceph-3em-4-carboxylic acid, syn isomer.

To the product obtained in Example 29 there were added 5 mi of methanol then 4 mi of a methanolic solution of sodium acetate. The mixture was concentrated to dryness under reduced pressure, then the residue was broken up in ethanol and vacuum-filtered to obtain 505 mg of the desired sodium salt.

Example 3 1: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2-(2-[1-methyl-tetrazol-5-yl-thio lethoxy-imino)-acetylaminol-ceph3-em4-carboxvlic acid.

Stage A: The syn isomer of 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4yl)-2-(2-[1-methyl-tetrazol-5-y lthiol-ethoxy-imino)-acetylaminoceph-3em4-carboxylic acid.

432 mg of the diethylamine salt of 3-acetoxymethy]-7-[2-(2tritylaminothiazol-4-yi)-2-(2-bromoethoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer were mixed with 64 mg of ll-methyl-5- 32 GB 2 101 117 A 32 mercapto-1,2,3,4-tetrazole and 2.2 m[ of anhydrous dimethy[formamide. The mixture was heated for 1 hour 50 minutes in a water bath at WC with agitation, then maintained for 1 hour 30 minutes at ambient temperature, and finally for 1 hour 10 minutes at WC. The mixture was cooled, diluted with 20 mI of water, agitated and the formed precipitate was vacuum- filtered to obtain 406 mg of a crude product.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2[1-methyl-tetrazol-5-yl-thio lethoxy-imino)-acetylamino-ceph-3em-4carboxylic acid.

406 mg of the product obtained in Stage A were dissolved in 3.2 m] of 66% aqueous formic acid, and the solution was heated in a water bath at WC for 15 minutes. The triphenylcarbinol formed was removed by vacuum-filtration and the aqueous formic acid was driven off by distillation under reduced pressure. The residue was taken up with ethanol and again concentrated to dryness. 3 m] of methanol were added to dissolve the residue and the solution was diluted with 30 m] of diethyl ether, and the precipitate formed was vacuum-filtered to obtain 201 mg of a crude produce which was dissolved in 1 mi of hot methanol and reprecipitated by adding 5 mi of ether. Vacuum-filtration yielded 171 mg of the desired product.

Analysis: C19H2107N9S3 Calculated: C% 39.10 1-1% 3.63 N % 21.6 S% 16.48 Found: 38.6 3.7 20.9 16 N.M.R. Spectrum (CID3)2SO P.P.M. 20 2.03: -O-C-CH3 1 0 4.33 (triplet): =N-O-CH2 Q = 6 Hz) 6.76: proton at 5-positionof thiazole 3.91 (single): =N-CH3 The diethylamine salt of 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)- 2-(2-bromoethoxy-imino)acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer, was prepared as follows:

0.79 of 3-acetoxymethyl-7-[2-(2-tritylamino-thiazol-4-yl)-2-(2bromoethoxy-imino)-ac etylaminol-ceph-3- 30 em-4-carboxylic acid was added to 5 CM3 of ethyl acetate. On total dissolution, 0.12 CM3 of diethylamine were added, and the mixture was agitated, vacuum-filtered and rinsed with ethyl acetate to obtain the desired salt after drying.

Example 32: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)2(2-azido-ethoxy-imino)- 35 acetylamino]-cep--3-em-4-carboxylic acid.

Stage A: The syn isomer of 2-(2-tritylaminothiazol-4-yl)-2-(2-azidoethoxy-imino)acetic acid.

For one hour in a water bath at 5WC, 24.8 g of sodium 2-(2-tritylaminothiazol-4-yl)-2-(2-bromoethoxy- imino)-acetate were agitated with 125 mi of dimethyiformamide and 7.5 g of tetramethylguanidine aziclicle.

After agitation for a further 45 minutes a clear brown solution was obtained, and this was cooled to ambient 40 temperature. 500 mi of water and 50 mi of 2N hydro chloric acid were added, and the mixture was vacuum-filtered, made into a paste three times with water, then ground up in methylene chloride. The filtrate was decanted, washed with water then dried. After concentration to 50 mi under reduced pressure 250 mI of diethyl ether were gently added, and the mixture was vacuum-filtered to obtain 15.58 g of the acid.

The sodium salt of 2-(2-bromoethoxy-imino)-2-(2-tritylaminothiazol-4-yi)acetic acid, syn isomer, was 45 prepared as follows:

6 g of sodium carbonate pellets were added to 280 CM3 of absolute ethanol. 28.2 g of ethyl 2-(2-bromo-ethoxy-imino)-2-(2-tritylamino-thiazol-4-yl)-acetate, syn isomer (described in Example 21) were added, and the mixture was agitated for 65 hours at ambient temperature. The sodium salt separated, and this was vacuum-filtered, washed with ethanol and dried in vacuo to obtain 29.82 g of the desired product as 50 a 12% solvate.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4yl)-2-(2-azido-ethoxy-imino)acetylaminol-ceph-3-em-4-carboxylic acid in the form of its diethylamine salt. 55 2.54 g of 7-amino-cephalosporanic acid, 25 m[ of anhydrous methylene chloride and 2.6 mi of triethylamine were agitated for 15 minutes at ambient temperature. 6.02 g of the 2-(2tritylamino-thiazol-4-yl)-2-(2-azido-ethoxy-imino)-acetate of 1-hydroxy-1 H-benzotriazole were added and the mixture was agitated for 45 hours at ambient temperature. 25 mI of water and 5 mi of N hydrochloric acid were added. The mixture was separated by decantation, and the organic phase was washed with water, dried and concentrated to dryness under reduced pressure. The residue was taken up 60 with 30 mi of ethyl acetate, and after dissolution, 2.4 m] of diethylamine were added. The mixture was cooled to O'C and vacuum-filtered to yield 5.9 g of the diethylamine salt.

The 2-(2-tritylamino-thiazol-4-yi)-2-(2-azido-ethoxy-imino)-acetate of 1 hydroxy-1 H-benzotriazole used in Stage B was prepared in the following way:

9.84 g of 2-(2-tritylamino-thiazol-4-yi)-2-(2-azido-ethoxy-imino)-acetic acid was mixed with 2.93 g of 65 33 GB 2 101 117 A 33 1-hydroxy-1H-benzotriazole, 4.86 g of dicyclohexylcarbodiimide and 130 mi of anhydrous methylene chloride, and the mixture was agitated for20 hours at ambient temperature. The dicyclohexylurea formed was removed byvacuum-filtration, and thefiltrate waswashed with waterto which had been added sodium bicarbonate, then with water alone. The washed filtrate was dried and concentrated to dryness under reduced pressure. The residue was taken up with ethyl acetate, cooled for half an hour at O'C and suction-dried to obtain 7.31 g of the desired product. A further 1.5 g of product was recovered from the mother liquors.

Stage C: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-azido-ethoxy-imino)acetyl- amino]-ceph-3-em-4-carboxylic acid.

For 15 minutes at WC 1.238 g of the product obtained in Stage B were agitated with 6 mi of 70% formic acid. The triphenylcarbinol formed was removed by vacuum-filtration and the filtrate was concentrated to dryness under reduced pressure. The residue was taken up with water, broken up, vacuum-filtered and dried to obtain 0.358 9 of the acid.

Example 33: Sodium salt of 3-acetoxymethyl-7-[2-(2-amino-thiazol-4-yl)-2(2-azido-ethoxy-imino)-acetyl- amino]-ceph-3-em-4carboxylic acid, syn isomer.

0.385 g of the acid obtained in Example 32 was mixed with 1 mi of a methanolic molar solution of sodium acetate. To the clear solution thus obtained were slowly added 5 m] of ethanol. The mixture was vacuum-filtered and rinsed with ethanol, then with diethyl etherto obtain 0.215 g of the sodium salt. 20 Analysis: C17H1707N8S2Na = 532.49 Calculated: C% 38.4 Found: 38.3 N.M.R. Spectrum flCD3)2SO P.P.M. 1.98 = -OC-CH3 11 0 4.18 (triplet): =N-O-CH2- J=5Hz 6.76: proton at 5 of thiazole.

H% 3.2 N% 21.0 S% 12.0 3.2 20.4 12.1 Example 34: The syn isomer of 3-acetoxymethyl-7-[2-(2amino-thiazol-4-yl)- 2-(2-amino-ethoxy-imino)acetyl-aminol-ce-h--3-em-4-carboxylic acid. Stage A: The syn isomer of 3acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-(2-aminoethyoxyimino)acetyl-aminol-ceph-3-em-4-carboxylic acid.

0.752 g of 3-acetoxym ethyl -7-[2-(2-trityl am i no-th iazo 1 -4-yi)-2-(2azido-eth oxy-i m in o)-acetyl am i no]-cep h-3em-4-carboxylic acid prepared in Example 32 were mixed with 4 mi of dimethy[formamide and 0.7 mi of anhydrous triethylamine. Hydrogen sulphide was bubbled into the mixture for 15 minutes, and 40 mi of water then 0.7 mi of acetic acid were added. After vacuum-filtration 0.707 g of a crude product were obtained.

Stage 8: The syn isomer of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-amino-ethoxy-imino)-acetyl- amino]-ceph-3-em-4-carboxylic acid. 1.054 g of the product obtained in Stage A were heated for 15 minutes at

WC in 5 mi of 70% formic acid. The triphenyl carbinol formed was removed by vacuum-filtration and the filtrate was concentrated under reduced pressure. The dry residue was taken up with water and the insoluble matter eliminated. The filtrate was concentrated to dryness and taken up with ethanol, broken up and vacuum-filtered to obtain 0.125 g of the acid.

Example 35.. Sodium salt of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-amino-ethoxy-imino)-acetylamino]-ceph-3-em-4-carboxylic acid, yn isomer.

The product obtained in Example 34 was dissolved in 0.2 mi of a molar solution of sodium bicarbonate. 0.4 mi of ethanol were carefully added, and the insoluble matter was removed by vacuum-filtration. The filtrate was concentrated to dryness under reduced pressure, and the residue was taken up with ethanol, crumbled and vacuum-filtered to obtain 0.047 9 of the desired salt.

Example 36: The syn isomer of 3-[(1-methyl- 1H-tetrazol-S-yl)-thiomethyll7-[2-(2-aminothlazol-4-yl)-2-(2azido-ethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid.

Stage A: Thesyn isomer of 3-[(1-methyP 1H-tetrazol-5-yl)-thiomethyll-7-[2(2-tritylaminothiazol-4-yl)-2-(2azido-ethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid.

For 15 minutes at ambient temperature 0.652 g of 7-amino-3-[(1 -methy]-1 H-tetrazo 1 -5-yi)-th io-m ethyl] ceph-3-em-carboxylic acid were agitated with 6.5 m] of methylene chloride and 0.56 mi of triethylamine. 1.29 g of the a-(2-azido-ethoxy-imino)-2-(2-tritylamino-thiazol-4-yi)-acetate of 1 -hydroxy-1 H-benzotriazole prepared65 34 GB 2 101 117 A 34 as described in Example 32, Stage B, were added, and the mixture was agitated for 20 hours at ambient temperature. 10 m[ of water and 3 mi of 2N hydrochloric acid were added. The mixture was separated by decanting and the organic phase was washed with water, dried and concnetrated to dryness under reduced pressure. The residue was taken up with 5 mi of ethyl acetate and 10 mi of diethyl ether were added. The mixture was agitated for half an hour at ambient temperature, and vacuum- filtered to obtain 1.416 g of a crude product.

Stage 8: The syn isomer of 3-[(1-methyl-1H-tetrazol5yl)-thiomethyll-7[2(2-aminothiazol-4-yl)-2(2-a zidoethoxyimino)-acetylaminol-ceph-3-em-4carboxylic acid.

For 15 minutes at WC 1.336 g of the product obtained in Stage A were agitated in 7 mI of formic acid. The triphenylcarbinol formed was removed by vacuum-filtration and the filtrate was concentrated to dryness under reduced pressure. The residue was taken up with water, broken up and vacuum-filtered to obtain 0.77 g of a crude product which was dissolved in a minimum amount of a 10% sodium bicarbonate solution. 0.07 g of active charcoal were added, and the mixture was vacuum-filtered. To the filtrate formic acid was added to give a pH of 2. The purified acid was vacuum-filtered to obtain 0.171 g of the desired product.

Example 37: Sodium salt of 3-[(1-methyl-1H-tetrazol-5-yl)-thiomethyll-7[2-(2-aminothiazol-4-yl)-2-(2-a zidoethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid syn isomer.

0.162 g of the purified acid obtained in Exam pie 36 were j7is--solved in 0.3 m I of a molar solution of sodium bicarbonate. 1 m[ of ethanol was added and the insoluble matter was vacuum-filtered off. The filtrate was 20 concentrated to dryness under reduced pressure, and the residue was taken up with ethanol, broken up and vacuum-filtered to obtain 0.09 g of the sodium salt.

Analysis: C17H1705N12S3Na: PM = 588.584 Calculated: C% 34.7 1-1% 2.9 Found: 34.5 3.2 N.M.R. Spectrum (CID3)2SO P.P.M. 3.9: N-CH3 6.76: proton in 5-position of thiazole. 9.35-9.48: -CONH- N% 28.5 S% 16.3 25.3 15.3 Example 38: The syn isomer of 3-[(1-methyl- 1H-tetrazol-5-yl)-thiomethyll7-[2-(2-aminothiazol-4-yl)-2-(2amino-ethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid.

StageA: 3-[(1-methyl- 1H-tetrazol-5-yl)-thiomethyll-7-[2-(2-tritylaminothiazo1-4-yl)-2-(2-amino-e thoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid.

3.91 g of sodium 3-[(1 -methyi-l H-tetrazol-4-yi)-thiomethyll-7-2-(2tritylamino-thiazol-4-y])-2-(2-azido- ethoxy-i m in o)-acetyl am in ol -ceph-3-em-4-ca rboxyl ate were dissolved in 39 m] of climethylformamide. 20 mi of water were slowly added, followed by 9.75 m] of triethylamine. Hydrogen sulphide was bubbled into the mixture. After 45 minutes, 3.9 mi of triethylamine were added and the bubbling in of hydrogen sulphide 40 continued for a further 15 minutes. The mixture was poured into a solution of dilute hydrochloric acid at 10'C, and the whole was heated for 15 minutes at WC under agitation, then cooled, and vacuum-filtered.

The precipitate was washed until neutral and rinsed with diethyl ether to obtain 2 g of the desired product.

The sodium 3-[(11 -methyl 1 H-tetrazo 1-5-yl)-th io-m ethyl 1-7-[2-(2trityla mino-th iazol-4-y])-2-(2-azido-ethoxy- im i no)-acetyl am i no]-ceph-3-em-4-ca rboxyl ate used at the beginning of the Example was prepared as 45 follows:

For one hour at ambient temperature 8.47 g of the syn isomer of 2-(2tritylaminothiazol-4-yl)-2-(2-azido- ethoxy-imino)-acetic acid obtained in Example 32 were agitated with 50 m] of methylene chloride and 1.93 g of dicyclohexylcarbodiimide. The dicyclohexylurea was removed by vacuumfiltration. The filtrate was added, over 20 minutes, to a mixture consisting of 3.07 g of 3-[(1- methylA H-tetrazo 1 -5-yl)-th io methyl] -7- 50 amino-ceph-3-em-4-carboxylic acid, 40 mi of anhydrous methylene chloride and 3.9 mi of triethylamine.

After agitation for a further hour the mixture was concentrated to dryness under reduced pressure. The residue was taken up with 50 mi of ethyl acetate at 2WC, acidified with 0. 2 m[ of acetic acid and vacuum-filtered. The filtrate was washed with N hydrochloric acid, then with water until neutral. The organic phase was dried and made up to a volume of 50 m[, then 1.7 mi of diethylamine were added. The salt of the starting acid crystallised, and was removed by vacuum-filtration. The filtrate was precipitated with 115 m[ of isopropyl ether. The precipitate was vacuum-filtered to obtain 7.62 g of the diethylamine salt. 5.9 g of this salt was taken up and dissolved under agitation in a mixture of 60 mi of water, 60 m[ of methiene chloride and 3.5 mi of 2N hydrochloric acid. The mixture was separated by decanting and the organic phase was washed with water dried and concentrated to dryness under reduced pressure. The residue was taken up with 20 mi of 60 isopropyl ether and vacuum-filtered to obtain 5.6 g of the free acid. This latter was dissolved in a mixture of 9.5 m[ of methanol and 6.7 mi of a 1 M methanol solution of sodium acetate. The sodium salt was precipitated with 27 m] of isopropanol containing 25% ethanol, then diluted with 270 m] of isopropanol to obtain 4.21 g of the desired product.

GB 2 101 117 A 35 Stage 8: The syn isomer of 3[(methyl1H-tetrazol-5-yl)-thiomethyll-7-[2(2-amino-thiazol-4-yl)-2-(2-aminoethoxy-imino)acetylaminol-ceph-3-em-4carboxylic acid.

2g of the product obtained in Stage A were heated to 40-45' under agitation with 5 mi of formic acid. 5 mi of water were added and the temperature maintained for 15 minutes. The mixture was then cooled and the triphenylcarbinol formed was vacuum-filtered off. The filtrate was concentrated to dryness under reduced pressure. The residue was taken up with 10 m] of ethanol, broken up, vacuum-filtered and rinsed with ethanol then with diethyl ether to obtain 1.36 g of a crude product which was taken up with 15 mi of 2N hydrochloric acid. The insoluble matter was vacuum-filtered off and the filtrate was then brought to pH 4 by the addition of 3 mi of an aqueous 1 M solution of lithium acetate, then an aqueous solution of lithium hydroxide. The mixture was vacuum-filtered and the filtrate was concentrated to dryness under reduced 10 pressure. The residue was taken up with 30 mi of ethanol, broken up and vacuum-filtered to obtain 515 mg. of the desired product. 133 mg of the product were recovered from the mother liquors. The two products were treated again in the same way as the crude product and 430 mg of a white product were isolated.

N.M.R. Spectrum (CD3)2SO P.P.M. 6.86: proton at 5-position of thiazole Example 39: Trifluoroacetate of 3-acetoxymethyl-7-12-(2-aminothiazol-4yl)-2-(2-iodoethoxy-imino)-acetyl amino]-ceph-3-em-4-carboxylic acid, syn isomer.

Stage A: Benzhydryl 3-acetoxymethyl-7-[2-(2-trityl-aminothiazol4-yl)-2-(2iodoethyoxy-imino)- acetylaminol-ceph-3-em-4-carboxylate, syn isomer.

Under an inert atmosphere 1.28 g of the syn isomer of 2-(2-iodoethoxyimino)2-(2-tritylamino-thiazol-4-yi)- acetic acid solvated with 1,2-dichloroethane (being in fact 1.09 g of pure product) were mixed with 1.45 g of the benzhydry] ester of 7-amino-cephalosporanic acid in 22 CM3 of anhydrous methylene chloride. The mixture was cooled in an ice bath and, drop by drop, 5 CM3 of a 0.5M solution (being 103 g/litre) of dicyciohexylcarbodiimide in methylene chloride were introduced. After agitation at Wto +WC for one hour thirty minutes, then for one hour at 2WC, the precipitated dicyclohexylurea was vacuum-filtered off, rinsing with methylene chloride. The filtrate and the washing waters were evaporated to dryness at a temperature lower than WC under reduced pressure to obtain 2.98 g of a product which was chromatographed on silica eluting with a 92-8 methylene chloride-ethyl acetate mixture. 1.12 g of white productwere obtained. 30 U. V. Spectrum (Ethanol/N hydrochloric acid) max. = 269 nm Ell = 209 lu LR. Spectrum (chloroform) = P-lactam: 1791 em-'; C=N-OR: 1042 em-'; C=C: 1638 em-' N.M.R. Spectrum (CDC13) proton at 5-position of thiazole: 6.75 p.p.m.

Stage 8: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-iodoethoxy-imino)acetylaminol-ceph-3-em-4-carboxylic acid, Iyn isomer.

960 mg of the product obtained in Stage A were introduced into 10CM3 of pure trifluoroacetic acid. The mixture was agitated for 3 minutes at ambient temperature, cooled for one minute in an ice bath and then precipitated by the addition of 100 cm3 of iced isopropyl ether. After agitation for 10 minutes at ambient temperature, the precipitate was vacuum-filtered rinsing it with isopropyl ether than with diethyl ether, and 45 dried under reduced pressure to obtain 460 mg of the desired product, m.p. #214'C.

Analysis: C171-118N507SI, CF3CO2H Calculated: N% 9.87 S% 9.04 Found: 9.7 9.2 U. V. Spectrum (Ethanol - - N hydrochloric acid) max. 262 nm E] = 290 10 N.M.R. Spectrum I(CID2)2SO/ proton at 5-position of thiazole: 6.83 p.p.m.

The syn isomer of 2-(2-iodoethoxyimino)-2-(2-tritylaminothiazol-4-yl)acetic acid used at the beginning of Example 39 was prepared as follows:

a) Ethyl2-(2-iodoethoxyimino)-2-(2-tritylaminothiazol-4-yl)-acetate, syn isomer.

6 9 of Ethyl 2-(2-bromoethoxyimino)-2-(2-tritylaminothiazol-4-yi)-acetate, syn isomer (prepared as in Example 1) were introduced into 60 em' of of methylethyiketone and 2.141 g of sodium iodide. The mixture was refluxed for an hour and 10 minutes, and then was evaporated under reduced pressure. The residue was taken up with 120 em' of methylene chloride and washed five times in 40 em' of water. Each washing was re-extracted with 2 em' of methylene chloride and the organic phase was dried and evaporated to dryness.65 36 GB 2 101 117 A The resin obtained was added to diethyl ether, and dried under reduced pressureto obtain 6.22 g of product. M. P. = 11 O'C.

E Analysis: C26H2203 N3S1 = 583.35 Calculated: N% 6.16 S% 4.70 Found: 5.9 4.8 U. V. Spectrum (Ethanol, N hydrochloric acid) 278 nm E] = 235 TO N.M.R. Spectrum (CDC13) proton at 5-position of thiazole: 6.58 p.p.m.

36 b) The syn isomer of 2-(2-iodoethoxyimino)-2-(2-tritylaminothiazol-4-yl)- acetic acid.

Under an inert atmosphere 6.7 g of the ethyl ester prepared in Stage a) were introduced into 5.5 CM3 Of dioxan and 44 CM3 of absolute ethanol. Drop by drop, 5.5 CM3 of a 2N solution of sodium carbonate were added, then 7 CM3 of absolute ethanol were added, and the mixture was agitated for one night at ambient temperature. The sodium salt thus formed was vacuum-filtered rinsing twice with 3 cm' of a 4-1 ethanol-dioxan solution and then formed into a paste with diethyl ether. The product obtained was treated in a separating funnel with 100 CM3 of water and 100 cm3 of chloroform. The pH was adjusted to 2 with normal 10 hydrochloric acid. The organic phase was decanted, washed with a saturated solution of sodium chloride, dried and evaporated under reduced pressure. The resin thus obtained was dissolved at 40'C in 35 CM3 Of dichloroethane. Crystallisation was initiated, and the solution was left at ambient temperature for 72 hours. The formed precipitate was vacuum- filtered, rinsed and dried to obtain 5.4 g of a white product solvated with dichloroethane (being 4.61 9 of pure product) m.p. = 161'C.

Example 40: N-[2-(2-[(2-carboxy-3-methyl-8-oxo-5-thia-l-azabicyclo[4.2. Oloct-2-en-7-yl)-a minol-l-[2-aminothiazol-4-yll-2-oxo-ethvl)-iminoxvethyll-pyridinium iodide trifluoroacetate double salt, syn isomer. Stage A: Benzhydryl 3-methy-7-[2-(2-tritylaminothiazol-4-yl)-2-(2bromoethoxyimino)-jc-etylamin ol-ceph-3- em-4-carboxylate, syn isomer.

Under an inert atmosphere 536 mg of the syn isomer of 2-(2bromoethoxyimino)-2-(2-tritylaminothiazol-4yl)-acetic acid were introduced into 380 mg of the benzhydryl ester of 7-amino-3desacetoxycephalosporanic acid and 6 cm 3 of anhydrous methylene chloride. The mixture was cooled in an ice bath and after 5 minutes 230 mg of dicyclohelyiccarbodiimide were added and rinsed with 2 c& of methylene chloride. The mixture was kept for two hours at Wto +5'C, then for one hour at ambient temperature, and vacuum-filtered washing 35 the insoluble matter three times with methylene chloride (and collecting 111 mg of product). The filtrate was brought to dryness and 1.02 g of resin were isolated. This product was purified by fixing it on 100 g of silica and eluting with a 17-3 benzene-ethyl acetate mixture. 548 mg of resin were obtained. Rf = 0. 27-0.28 with preceding eluant.

N.M.R. Spectrum proton at 5-position of thiazole: 6.75 p.p.m. -CH2-Br triplet centred on 3.58 p.p.m. J = 7 Hz.

Stage B: N-[2-(2-[(2-diphenylmethylcarboxy-3-methyl-8-oxo-5-thialazabicyclo[4.2.Olo ct-2-en-7-yl)-aminol1-[2-tritylaminothiazol-4-yll-2-oxo-ethyl)-iminooxy-ethyll-pyridinium iodide, syn isomer.

Under an inert atmosphere 500 mg of the product obtained in Stage A were introduced into 115 mg of pyridine iodhydrate in 5 CM3 of pyridine. The mixture was heated at WC for 15 hours then evaporated under reduced pressure at a temperature lower than WC. The residue was taken up with methanol and evaporated four times in succession to drive off residual pyridine, then dried under reduced pressure to obtain 620 mg Of 50 a crude product. This crude product was purified by chromatography on silica, eluting with a 85-15 chloroform-methanol mixture, to obtain 348 mg of a purer product in the form of resin.

N.M.R. Spectrum (CD3)2SO proton at 5-position of thiazole: 6.81 p.p.m.

Stage C: N-[2-(2-[(2-carboxy-3-methyl-8-oxo-5-thia- 1-azabicyclo[4.2. 0]oct-2-en7-yl)aminol-l-[2-aminothiazol-4-yll-2-oxo-ethyl)iminooxylethyll-pyridinium iodide trifluoroacetate double salt, syn isomer.

300 mg of the product obtained in Stage B were added to 3 CM3 of pure trifluoroacetic acid. The solution was agitated for 3 minutes at ambient temperature and cooled for 20 seconds in an ice bath. A precipitate was formed by adding 40 CM3 of an iced 1 -1 mixture of isopropyl ether and petroleum spirit B (boiling point between 65 and 7WC). The precipitate was vacuum-filtered, rinsed with isopropyl ether, then with diethyl ether and dried to obtain 152 mg of a powder, m.p. 222'C. Rf = 0.05 (acetic acid-ethyl acetate-water 70-35- 10) 37 GB 2 101 117 A 37 U.V. Spectrum (Ethanol; N hydrochloric acid) 10 max. = 260 nm Ell = 355 I.R. Spectrum P-factam 1768 cm-1 O=N-OR 1038cm-1 N.M.R. Spectrum /(C133)2SO/ jo proton at 5-position of thiazole = 6.76 p.p.m.

Example 4 1: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(2-dimethylaminoethoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer. Stage A: Hydriodic of 2-(2-dimethylaminoethoxyimino)-2(2-tritylaminothiazol-4-yl)-acetic acid, syn isomer.

At ambient temperature 120 cm 3 of a 9:1 mixture of chloroform and dimethylamine and 10 mg of the syn 15 isomer of 2-(2-iodoethoxyimino)-2-(2-tritylaminothiazol-4-yi)-acetic acid were agitated for about 3 hours. The mixture was evaporated to dryness without the temperature exceeding 4WC and the residue was taken up with 100 CM3 of isopropyl ether, ground, agitated for 15 minutes at ambient temperature, vacuum-filtered and rinsed. The residue was taken up with 50 cm 3 of acetone, refluxed for 5 minutes then vacuum-filtered at ambient temperature, dried and the solvents removed by evaporation under reduced pressure. 20 9.33 g of product were obtained which were taken up with 46 CM3 of water, agitated, then vacuum-filtered and rinsed. The residue was taken up with 80 CM3 of acetone, agitated, vacuum-filtered and dried under reduced pressure to give the desired product.

M.p. = 208-210'C (decomposition).

N.M.R. Spectrum (D20 + C5DO: p.p.m. Triplet centred on 4.55 Q = 5 Hz): =NO-CH2 single at 6.98 proton at 5-position of thiazoleT single at 7.33 C(Ph)3 U.V.Spectrtipm(Ethanol, N hydrochloric acid) 10 infl. 270 nm E' = 271 max. 275 nm Ell = 280 P- = 14,000 infl. 284 nmE] = 260 Stage 8: Benzhydryl3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-(2dimethylami noethoxy-imino)acetyl-aminol-ceph-3-em-4-carboxylate, syn isomer. 1 g of the acid prepared in Stage A was dissolved in a mixture of 15 cm3 of chloroform and 1.5 CM3 Of 40 methanol and 304 mg of triethylamine hydrochloride were added. The mixture was heated for 5 minutes at 40 MC then evaporated to dryness. The residue was taken up in 15 CM3 of chloroform, and brought to 00 to +50C by means of an ice bath. Drop by drop, 2 CM3 Of a solution of pivaloyl chloride (prepared by making 1.25 g of pivaloyl chloride up to 10 cm' with chloroform) were added. The mixture was leftto return to ambient temperature. 45 At the end of two hours, 1.1 g of benzhydryl 7-amino-cephalosporanate were added. At the end of one hour thirty minutes the mixture was evaporated to dryness and chromatographed on silica eluting with chloroform containing 5% methanol, to obtain 430 mg of the desired product.

Stage C: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-dimethylaminoethoxy-imino)- acetylaminojceph-3-em-4-carboxylic acid, syn isomer.

A mixture of 100 mg of the product obtained in Stage B in 1 cm 3 of trifluoroacetic acid was agitated for 3 minutes at ambient temperature. A precipitate was formed by adding ether, which was taken up with 0.2 cml of methanol. Again a precipitate was formed with 2 CM3 of ether. This was formed into a paste with chloroform then vacuum-filtered and rinsed with chloroform, then with ether, to obtain 40 mg of the desired product. M.p. = 23WC.

N.M.R. Spectrum (CD3)2SO (P.P.M.) single at 6.8: proton at 5-position of thlazole U. V. Spectrum Ethanol max. 234 nm Ell = 305 E = 15,600 infl. 254 nm E' = 247 E = 12,650 infl. 296 nm Ell = 103 e = 5,300 Ethanol; h hydrochloric acid 38 GB 2 101 117 A max. 260 nm Ell = 279 E = 14,300 infl. 276 nm Ell = 243.

38 Example 42: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(2-pyridylethoxyimino)- acetylaminolceph3-em-4-carboxylic acid, syn isomer (in the form of internalpyridinium salt). Stage A: Syn isomer of2-(2-tritylamino-thiazol4- yl)-2-(2-pyridyl-ethoxy-imino)-acetic acid (in the form of intemalpyridinium salt).

g of the syn isomer of 2-(2-iodo-ethoxy-imino)-2-(2-tritylaminothiazol-4yi)-acetic acid solvated with dichloroethane, (corresponding to 4.27 g pure compound and obtained as described in Example 39) were heated to 60'Cfor 24 hours in 30 CM3 of pyridine then left for 56 hours at ambient temperature. The 10 precipitate thus formed was vacuum-filtered, rinsed with pyridine then with ether and dried to obtain 1.66 g of product, m.p. = 25WC (decomposition).

N.M.R. Spectrum (CD3COCD3, D20 = 1-1) 15 6.8 p.p.m.: H5 of thiazole 7.5 to 8 p.p.m.: pyridyl proton 4.58 to 5.08 p.p.m.: = N-O-CH2-CH2 U, V. Spectrum (EtOH-HCI N/10) 20 Max. 260 nm Ell = 324 I.R. Spectrum (nujol) C0261639 em-' C = C 1583 em-' 25 C = N 1523 em-' Stage 8: Benzhydryl 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-(2pyridylethoxyimino)ac etylamino]-ceph-3-em-4-carboxylate iodide, syn isomer.

540 mg of the product obtained in Stage A were agitated for 20 minutes at ambient temperture with 210 mg 30 of pyridine hydriodide, 420 mg of dicyclohexylcarbodiimide, 350 mg of benzhydryl 7-amino-3-acetoxymethyl-ceph-3-em-4-ca rboxyl ate and 5 CM3 of anhydrous dimethylformamicle. The dicyclohexylurea precipitate formed is vacuum-filtered and rinsed with dimethylformamide, and the filtrate was poured into CM3 of diethyl ether, where it formed a gum. The whole was agitated for 5 minutes at ambient temperature, decanted and the supernatent liquid was removed. The gum was taken up and ground in 60 CM3 of diethyl ether. The precipitate formed was vacuum-filtered to obtain 754 mg of a crude product. This 35 was purified by agitation for 20 minutes with 1.6 g of magnesium silicate in 7.5 CM3 of dichloroethane. The insoluble matter was vacuum-filtered off and washed with 0.5 CM3 of 1,2clichloroethane, then the filtrate was evaporated to dryness to obtain 368 mg of the desired product.

Stage C: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(2-pyridylethoxy-imino)-acetyl- 40 amino]-ceph3-em-4-carboxylic acid, syn isomer.

The 368 mg of product obtained in Stage B were agitated for 3 minutes at ambient temperature in 3 CM3 Of pure trif 1 uoroacetic acid. The insoluble matter was rapidly vacuum-f iltered off. A precipitate was formed in the filtrate by adding 40 CM3 of diethyl ether and agitating for 5 minutes. The precipitate was vacuum-filtered and rinsed with diethyl ether to form a hygroscopic product which was taken up with 0.35 CM3 of methanol 45 and precipiated with 4 CM3 of diethyi ether. The reformed precipitate was vacuum-filtered and rinsed with diethyl ether to obtain 140 mg of the desired product. M.p. = 205'C (decomposition).

N.M.R. Spectrum (DIVIS0) 50 2.06 p. p. m.: 0Ac 6.86 p.p.m.: H5 thiazole 8.9-9.1 p.p.m.: H2 and H6 8.55 to 8.78 p. p. m. H4 8.03 to 8.26 p.p.m. H3 and H5 55 U. V. Spectrum (ROH, HCI NA 0) Max. 260 260 nm E1 = 318 I.R. Spectrum (nujol) P lactam 1777 em-' 0Ac 1740 em-' C02 1633 em-' C=NO-R 1037 em-' 39 GB 2 101 117 A 39 Example 43: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(2-imidazol-l-yl-ethoxyimino)-acetylaminol-ceph-3-em-4-carboxylic acid, Hn isomer. Stage A: The syn isomer of 2-(2-tritylaminothiazol-4-yl)2-(2-imidazol-l-yl-ethoxyimino)-acetic acid (in the form of intertalimidazolium salt).

3 g of the syn isomer of 2-(2-tritylaminothiazol-4-yl)-2-(2iodoethoxyimino)-acetic acid, solvated with 1,2-dichloroethane, (corresponding to 2.56 g pure compound) were agitated for 3 hours at ambient temperature with 4.2 g ofirnidazole and 10 CM3 Of dimethylacetamide. A further 10CM3 of dimethylacetamide were added and the agitation continued for 40 hours at ambient temperature. The mixture was poured into 200 CM3 of isopropyl ether, agitated for 30 minutes, left to separate and then the supernatent liquid was jo removed. The gum thus obtained was taken up by grinding in 200 CM3 of Isopropyl ether, and again in 300 cm 3 of diethyl ether, then agitated for 30 minutes, vacuum-filtered, rinsed with diethyl ether, taken up with 30 cm 3 of acetone, agitated for one hour, vacuum-filtered, rinsed with acetone then with diethyl ether and dried to obtain 1.2 g of the desired product. M.p. = 28WC (decomposition).

N.M.R. Spectrum (DMSO) 4.35 p.p.m. N-O-CH2-CH2- N-- 6.8 p.p.m. H5 of thiazole 7.82 p. p. m.

U. V. Spectrum (ROH, HCI N/1 0) Max. 277 nm Ell = 259 I.R. Spectrum (nujol) C02- 1614 cm aromatic 1492 cm-1 heterocycle 1527 cm-1 h N 11 Stage 8: Benzhydryl3-acetoxymethyl-7-[2-trityl-aminothiazol-4-yl)-2-(2imidazo1-1-yl-e thoxy-imino)- acetylaminol-ceph-3-em4-carboxylate.

780 mg of the product obtained in State A were agitated vigorously in a bath of iced water with 315 mg of pyridine hydriodide, 630 mg of dicyclohexylcarbodiimide, 600 mg of benzhydryl 7-amino-3-acetoxymethyiceph-3-em-4-ca rboxyl ate and 6 CM3 of anhydrous dimethylformamide. Dicyclohexyl urea precipitated. The mixture was taken to ambient temperature and the agitation was continued for 20 minutes. The insoluble matter was removed by vacuum-filtration and rinsed with dimethylformamide. The filtrate was treated with CM3 of diethyl ether to form a precipitate, then agitated for 20 minutes and left to separate. The supernatent liquid was removed and the gum obtained was taken up and triturated in 100 CM3 of diethyl 40 ether. This mixture was agitated for 10 minutes at ambient temperature, vacuum-filtered, rinsed with diethyl ether and dried to obtain 1.3 g of a product which was chromatographed on silica, eluting with a 8-1-0.5 mixture of ethyl acetate-eth an o 1 -water to obtain 332 mg of the desired product.

N.M.R. Spectrum (CDC13) 2 p.p.m.: 0Ac 6.75 p.p.m.: H5 syn thiazole I.R. Spectrum (CHC13) P lactam: 1788 cm-1 ester and 0Ac: 1759 cm-1 heterocycles A525cm-1 Stage C: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2- (2-imidazol- l-ylethoxy-imino)-acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer.

230 mg of the product obtained in Stage B were agitated for 3 minutes at ambient tem peratu re with 2 CM3 of pure trifluoroacetic acid. The formed solution was poured into a 1 -1 mixture of isopropyl ether-diethyl ether and agitated for 20 minutes at ambient temperature. The formed precipitate was rinsed with diethyl ether, then taken up in 0.4 CM3 of methanol and again precipitated with 4 CM3 of diethyl ether, agitated for 10 minutes at ambient temperature, vacuum-filtered, rinsed with diethyl ether and dried to obtain 140 mg of the desired product. M.P. = 205'C (decomposition).

GB 2 101 117 A N.M.R. Spectrum QMS0) 2.03 p. p. m.: OAC 6.8 p.p.m.: H5 syn thiazole 7.66 and 7.71 p.p.m.: H4 and H5) imidazole 8.95 p.p.m.: H2 5 U. V. Spectrum (EtOH, NCI N/1 0) Max. 260 nm Ell = 271.

Example 44: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(ethoxycarbonyloxy-imino)acetyl-aminol-ceph-3-em-4-carboxylic acid, syn isomer. StageA: Diethylamine salt of 3-acetoxymethyl-7-[2-(2tritylaminothiazol-4-yl)-2-(ethoxycarbonyloxy-imino) acetylaminol-ceph-3em-4-carboxylic acid, syn isomer.

Under agitation and in an inert atmosphere 1.67 9 of the syn isomer of 3acetyoxymethyi-7-[2-(2- 15 tritylaminothiazol-4-yi)-2-hydroxyimino-acetylaminol-ceph-3-em-4carboxylic acid were dissolved in 25 CM3 of methylene chloride and 0.37 cm3 of pyridine. The solution was cooled in an ice bath, and then over five minutes 2.7 CM3 of a molar solution of ethyl chloroformate in methylene chloride were added. After 10 minutes and still at Wto +WC, 20 CM3 of water and 2.7 CM3 of N hydrochloric acid were added. This mixture was agitated, then separated. The organic phase was washed with water until neutral and then dried. The 20 solvent was distilled off under reduced pressure and the residue was taken up with 10 CM3 of ethyl acetate, then 0.23 CM3 of diethylamine were added. The diethylamine salt was precipitated by adding 10 CM3 Of isopropyl ether. The salt was vacuum-filtered and rinsed with a 1 -1 mixture of isopropyl ether-ethyl acetate, then with isopropyl ether, to obtain 1.60 g of a product which was dissolved in 5 cm 3 of methylene chloride and 10 c& of ethyl acetate. The solution was concentrated under reduced pressure to obtain a final volume 25 ofabout6 CM3 and then diluted with 5 CM3 of isopropyl ether, vacuum- filtered and rinsed as previously to obtain 1.47 g of the desired product.

N.M.R. Spectrum (CDC[3) 2.03 p.p.m. 0Ac 6.95 p.p.m. H5 of thiazole 7.66 p.p.m. trityl 4.13 - 4.25 - 4.36 - 4.48 p. p. m.: CH2 Of C02 Et LR.Spectrum (CHC13) P lactam 1781 cm-1 0Ac 1740 cm-1 amide 1694 cm-1 C021634 cm - 1 Stage 8: Trifluoroacetate of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-ylj-2(ethoxycarbonyloxy-iminoj-acety l- amino]-ceph-3-em-4-carboxylic acid, syn isomer.

1.43 g of the product obtained in Stage Awere agitated for 20 minutes at ambient temperature with 5.7 cm 3 of trifluoroacetic acid. 57 CM3 of isopropyl ether were rapidly added and the mixture was agitated for a further 15 minutes, then vacuum-filtered and rinsed with isopropyl ether to obtain 1.04 g of a crude product 45 which was dissolved in 4 CM3 of acetone containing 1% water. To the solution was added 12 cm 3 of diethyl etherto precipitate 0.69 g of the desired product. A further 0.11 g of identical product were recovered from the mother liquors.

Example 45: Sodium salt of 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2(ethoxycarbonyloxy-imino) 50 acetylamino]-ceph-3-em-4-carboxylic acid, syn isomer.

0.8 g of the product obtained in Example 4-were dissolved in 4 CM3 of a 1 M methanolic solution of sodium acetate and 2 CM3 of methanol. The solution was treated with active charcoal and diluted with 20 cm 3 of anhydrous ethanol then concentrated to a final volume of 10 CM3 by distilling under reduced pressure at WC maximum. The precipitate obtained was vacuum-filtered, rinsed with ethanol and then with etherto obtain 55 420 mg of product.

580 mg of product prepared in this way were taken up with 5.8 CM3 of methanol and while agitating 1.2 cm 3 of ethanol were slowly added. The precipitate was vacuum-filtered off and the filtrate was diluted with 10cm 3 of ethanol then concentrated by distilling under reduced pressure at WC maximum to a final volume of 5 CM3. This was vacuum-filtered and rinsed with ether to obtain 460 mg of product which was treated 60 again in the same mannerto obtain 420 mg of the desired product.

41 N.M.R. Spectrum (DMSO) 1.13-1.25-1.36p.p.m.

4.03-4.164.28-4.4p.p.m.) 1.98 p.p.m.: 0Ac 7.03 p.p.m.: H5 of thiazole U. V. Spectrum (EtOH/HCI N/1 0) 10 Max. 259 nm Ell = 324 F 17,350 Infl. 278 nm E] = 252 ) C02Et I.R. Spectrum (nujol) Plactarn:1763cm-10Ac:1726cm-1C02:1609cm C=N-O: 1038cm-1 GB 2 101 117 A 41 Example 46:3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2-morpholinoethoxy-imino)-a cetylaminol-ceph- 3em-4-carboxylic acid, syn isomer in the form of internal salt.

Stage A: 2-(2-tritylaminothiazol-4-yl)-2-(2-morpholino-ethoxy-imino)acetic acid, syn isomer in the form of internalsalt.

2 g of the syn isomer of 2-(2-tritylaminothiazol-4-yi)-2-(2-iodoethoxyimino)-acetic acid solvated with 1,2-dichloroethane (corresponding to 1.71 g of pure compound) were dissolved in 7 CM3 of morpholine. The solution was agitated for one hour at ambient temperature then concentrated in a current of nitrogen, and taken up with 30 em' of ethyl acetate, agitated fortwenty minutes at ambient temperature and vacuum-filtered, rinsing with ethyl acetate. The product was agitated for fifteen minutes in 15 CM3 Of 25 1,3-dimethoxypropane then vacuum-filtered and rinsed with the same solvent. The product was then taken up with 15 CM3 of ether, agitated for 15 minutes, vacuum-filtered and rinsed with diethyl ether to obtain 2.3 g of product, m.p. = 18WC (decomposition), which was used for the following stage in that form.

mg of the product was purified by recrystallisation from 0.5 em 3 of ethanol to obtain 46 mg of recrystallised product, m.p.: 182-184C (decomposition).

N.M.R. Spectrum (CDC13) 7.33 p.p.m.: trityl 6.75 p.p.m.: H5 syn thiazole U. V. Spectrum (ROH, HCl N/1 0) Max. 276 nm E] = 172 I.R. Spectrum (CHC13) 40 C02- and aromatics 1606-1529 and 1495cm-1; -NH 3399cm-1 Stage 8: BenAydryl, 3-acetoxymethyl 7-[2-(2-trityl-aminothiazol-4-yl)-2(2-morpholino-ethoxy-iminol- acetylaminol-ceph-3-em-4-carboxylate hydriodide, syn isomer.

1 g of the product obtained in Stage A, 0.38 9 of pyridine hydroiodide, 0. 63 g of dicyclohexylcarbodiimide and 0.60 g of benzhydryl 3-acetoxymethyl-7-a m i no-ce ph-3-em-4-ca rboxyl ate were dissolved in 5 CM3 Of 45 anhydrous dimethylformamide. The solution was agitated for 30 minutes at ambient temperature. The dicyclohexylurea thus formed was removed by vacuum-filtration; 100 c& of diethyl ether were added to the filtrate, and the mixture was agitated for ten minutes, vacuum-filtered, rinsed with diethyl ether and chromatographed on silica eluting with a 7-1 ethyl acetate-ethanol mixture to obtain 0.524 9 of product. M.p.

50:167'C (decomposition).

N.M.R. Spectrum (CDC13) 2.03 p. p. m.: 0Ac 3.72 p.p.m. CH20 6.75 p.p.m. H5 syn thiazole U. V. Spectrum (EtoH, H0 N11 0) Max. 268 nm Ell = 165 E= 15,900 LR. Spectrum (CHC13) P lactam 1791 em-' ester and 0Ac 1740 em-' amide 1678 em-' 42 GB 2 101 117 A Stage C: 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2-morpholinoethoxyimino)-acet ylaminol-ceph-3em-4-carboxylic acid, syn isomer, in the form of intemal salt.

0.42 g of the product obtained in Stage B were agitated for two minutes at ambient temperature with 3 cml of trifluoroacetic acid. The insoluble matter was vacuum-filtered off and rinsed with trifluoroacetic acid. The filtrate was collected in 30 CM3 of diethyl ether and agitated for 15 minutes at ambient temperature. The precipitate formed was vacuumfiltered, rinsed with ether and dissolved in 0.5 cm' of methanol. Then it was again precipitated with 5 CM3 of diethyl ether. After agitation for 5 minutes, the precipitate is vacuum-filtered off to obtain 208 mg of product, m.p. = 212-214'C (decomposition), which was salified in various ways, and in particular by trifluoroacetic acid.

N.M.R. Spectrum (DMSO) 2.05 p.p.m.: 0Ac 3.17 to 4.66 p.p.m.: CH2- Nand CH20 6.85 p.p.m.: H5 syn thiazole U. V. Spectrum (ROH, HCl N/10) Max. 260 nm Ell = 253 I.R. Spectrum (CHC13) 20 P lactam 1797 cm-1 C02- 1634 cm-1 amide 1667 cm- 1 42 Example 47: 3-acetoxymethyl-7-[2-(2-aminothiazol-4yl)-2-(2-[4-methyl- piperazinl-yll-ethoxy-imino)-acetyl- amino]-ceph-3-em-4-carboxylic acid, yn isomer, in the form of intemal salt (trifluoroacetate).

Stage A: 2-(2-tritylaminothiazol-4-yl)-2-(2-[4-methyl-piperazin- l-yllethoxy-imino)-acetic acid, in the form of intemalsalt.

2.22 g of N-methyl-piperazine in 15 cm 3 of dioxan and 3.325 g of the syn isomer of 2-(2-tritylaminothiazol 4-yl)-2-(2-iodoethoxy-iminol-acetic acid solvated with dichloroethane, (corresponding to 2.84 g pure compound and prepared as in Example 39) were agitated vigorously for 16 hours at ambient temperature. 30 The N-methyl piperazine hydriodide thus formed was vacuum-filtered off and the filtrate was concentrated under reduced pressure to a volume of about 5 CM3 then 200 CM3 of isopropyl ether were added and the mixture was agitated for 30 minutes. The mixture was vacuum-filtered and the product was taken up with 10 cm 3 of climethoxypropane, agitated for thirty minutes, vacuum-filtered, taken up with 24 CM3 of water, agitated for ten minutes, then again vacuum-filtered. The insoluble matter was agitated fortwo hours in 100 35 cm 3 of 1-1 isopropyl ether-diethyl ether mixture, then vacuum-filtered off and recrystallised from ethanol at 100'to obtain 1.47 g of product. M.p.: 22WC (decomposition).

N.M.R. Spectrum (CDC]3) 2.52p.p.m.:-NCH3 3.0 p.p.m. CH2N 6.7 p.p.m. H5 of thiazole U. V. Spectrum (EtOH/HCI N/1 0) Max. 277 nm Ell = 257 c 14,300 I.R. Spectrum (nujol) C02- 1602 cm-1 heterocycle 1529 cm-1 absenceOfCO2H Stage 8: Benzhydryl 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)2-(2[4-methyl-piperazin- l-ylj-ethoxy imino)-acetyllaminol-ceph-3-em-4-carboxylate, syn isomer.

560 mg of product obtained in Stage A was heated to WC for ten minutes with 140 mg of triethylamine hydrochloride in 5 cm' of chloroform and 5 cm3 of methanol. The solvents were evaporated off under reduced pressure, and the residue was taken up with 20 cm' of methylene chloride then cooled to -20'C.

Over ten minutes 1 CM3 of a 12.5% solution of pivaloyl chloride in chloromethylene was added and the whole agitated for an hour at ambient temperature. The solution was cooled to - 10'C and, in one go, 350 mg of benzhydryl 7-amino-3-acetoxymethyl-ceph-3-em-4-carboxylate were added. The mixture was agitated for three hours, then concentrated to dryness under reduced pressure at a maximum of WC. The residue was 60 taken up with 50 CM3 of an 8-2 benzene-ethyl acetate mixture, and the insoluble matter was eliminated. The solution was evaporated to dryness under reduced pressure, and the residue taken up with 25 CM3 Of 1,2-clichloroethane. 1.5 g of activated magnesium silicate were added and the mixture was agitated for 30 minutes, then vacuum-filtered and rinsed with 1,2-dichloroethane. The filtrate was evaporated under reduced pressure at3WC maximum to obtain 870 mg of white resin.

43 GB 2 101 117 A 43 N.M.R. Spectrum (CDC13) 2.02 - 2.03 p. p. m.: 0Ac 6.87 p.p.m. H5 of thiazole 2.45 p.p.m. N -CH3 U. V. Spectrum (ROH, HO N/1 0) Max. 267-268 nm: 14,700.

Stage C: 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(2-[4-methylpiperazin-l-yll-et hoxy-imino)- acetylaminol-ceph-3-em-4-carboxylic acid, syn isomer, in the form of internal salt (trifluoroacetate) 524 mg of the product obtained in Stage B were dissolved in 3 cm 3 of pure trifluoroacetic acid and the solution agitated for one minute at ambient temperature. A precipitate was formed by adding 30 cm3 of isopropyl ether and the whole was agitated for a further 5 minutes, then vacuum-filtered to obtain 320 mg of a hygroscopic product. This was dissolved in 1 cml of methanol and caused to precipitate by adding 10CM3 of diethyl ether. The mixture was agitated for 5 minutes, vacuum-filtered, rinsed with diethyl ether, made 15 into a paste with chloroform then with ether to yield 220 mg of a salified product (trifluoroacetate) M.p. 24WC.

N.M.R. Spectrum (DMSO) 2.03 p.p.m.: 0Ac 2.75 to 3.67 p.p.m.: CH2N 6.78 p.p.m.: H5 of thiazole U. V. Spectrum (ROH, HCI N/1 0) Max. 262 nm: 14,200 I.R. Spectrum (nujol) 0 lactam 1773 cm-1 0Ac 1726 cm - C020 1627 cm C=NO=1035 cm-1 Example 48: Bis trifluoroacetate salt of 3-acetoxymethyl-7[2-(2- aminothiazol-4-yl)-2-(2-aminoethoxy-iminoacetyl-aminol-ceph-3-em-4-carboxylic acid, syn isomer Stage A: 2-(2-tritylaminoethoxyimino)-2-(2-tritylamino-thiazol-4-yl)- acetic acid, syn isomer A mixture of 75 g of tritylamine, 50 g of ethyl and 100 CM3 of dimethyl sulphoxide was placed under argon, and the suspension was agitated for 82 hours at WC. ltwas then cooled to ambient temperature and caused to precipitate with 10 volumes of water. The precipitate was vacuum- filtered, washed with water, and then put into solution in one litre of chloroform and washed with water, then with a saturated solution of sodium chloride. The solution was dried and evaporated under reduced pressure at a temperature lower than 400C. 40 The crude product was taken up with a mixture of 320 cml of dioxan, 2 litres of absolute ethanol and 200 CM3 of 2N sodium carbonate, and agitated for 24 hours at ambient temperature, then evaporated to dryness under reduced pressure at a temperature lower than or equal to 4WC.

This product was pasted 5 times with 60 CM3 of a 1-7 dioxan-methanol mixture. The product was taken up with a mixture of a litre of chloroform and a litre of water.

Under agitation normal hydrochloric acid was added to adjust the pH to 2. The organic phase was separated, washed with water and then with a saturated solution of sodium chloride, dried and evaporated to dryness under reduced pressure at a temperature lower than 40'C. The product thus obtained was suspended in 300 CM3 of 1,2-dichloroethane and heated for 15 minutes at WC. Then it was vacuum-filtered after returning to ambient temperature under light vacuum, and rinsed with dichloroethane then with isopropyl ether and finally with diethyl ether. After being dried to constant weight 40 9 of powder were obtained. M.p. = 176'C.

Stage C. DiphenyImethyl 3-acetoxymethyl-7-[2-(2-tritylaminoethoxy-imino)2-(2-tritylamino-thiazol-4-y l)55 acetyl-aminol-ceph-3-em-4-carboxylate, syn isomer 11.55 g of the activated ester product prepared in Stage B were mixed with the equivalent of 7.65 g of the diphenyimethyl ester of 7-amino cephalosphoranic acid in 75 CM3 of methylene chloride. The mixture was agitated for 18 hours at ambient temperature, evaporated under reduced pressure and chromatographed on silica and under pressure, eluting with an 85-15 benzene-ethyl acetate mixture to obtain 9.1 g of the desired product.

Stage D: Bis trifluoroacetate salt of 3-acetoxymethyl-7-[2-(2aminothiazol-4-yl)-2-(2-aminoethoxy-imino)acetyl-aminol-ceph-3-em-4carboxylic acid, syn isomer Diphenyimethyl 3-acetoxymethyi-7-[2-(2-tritylamino-thiazol-4-yi)-2-(2tritylaminoethoxy-imi no)- acetylaminol-ceph-3-em-4-carboxylate prepared in Stage C were introduced into 1.8 CM3 of pure 44 GB 2 101 117 A 44 trifluoroacetic acid. The yellow solution thus obtained was agitated for3 minutes at ambient temperature, then underan inert atmosphere in a bath of iced waterwhen 18 CM3 of isopropyl ether were rapidly added. The mixture was agitated for 10 minutes, vacuum-filtered, rinsed with isopropyl etherthen with diethyl ether and dried to obtain 100 mg of a white powder. M.p. = about 21 O'C (decomposition).

Claims (31)

1. A compound of the general formula:

NHR16 10 5"1 N C02H 15 N ORi wherein R16 represents a protecting g rou p for the amino radical and Wi represents: a radical -CX- Wi3 in which Wi3 represents an alkyl radical having from 2 to 4 carbon atoms, an alkoxy group having from 1 to 4 carbon atoms, a phenyl radical or a radical -(CH2)r,-NR"2R"3, in which n represents 0 or an integer of from 1 to 4 and W2 and W3, which may be the same or different, each represent a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms with the proviso that W2 and W3 do not both represent hydrogen when n is 25 zero, or W2 and W3 together with the intervening nitrogen atom represent a phthalimido, piperidino or morpholino group and X is oxygen or sulphur; a radical -CX-NI-12; a radical -CHR'4-C02A"', in which W4 represents a phenyl or cyano radical and A- represents an easily-cleavable ester group as hereinbefore defined, a radical v ', 35 0 a radical - (CH2)n-- W5, in which n' is a n integer of f rom 1 to 4 and W5 represents an alkoxy radical having from 1 to 4 carbon atoms, a radical Alk-S(O),- in which Alk represents an alkyl radical containing from 1 to 4 carbon atoms and ns is 0, 1 or 2, or an acyl radical having from 2 to 4 carbon atoms; a radical -(CH2),2-CN in which n2 is 2, 3 or 4; a radical - (CH2)n'-CX'-NH2, in which X' represents a sulphur atom or, when n' is an integer otherthan 1, an oxygen atom, a radical 45 in which W7 represents a methyl or amino radical; a radical (CH2 N 55 a radical 60 a radical -(CH2)'-R5a in which R5, isa halogen atom, ora radical -S- Rarinwhich Rarrepresentsa phenyl radical or a 5- or 6-membered aromatic heterocyclic radical containing from 1 to 4 heteroatoms selected from sulphur, nitrogen and oxygen, the phenyl and heterocyclic radicals being optionally substituted by one or more radicals selected from amino, nitro and cyano radicals and alkyl radicals having from 1 to 4 carbon 65 atoms; GB 2 101 117 A 45 a radical Rb which is a radical -(CH2),,-R5b in which R5b represents an imidazolyl, morpholinyl or N-alkyl piperazinyl radical, the Walkyl radical containing from 1 to 4 carbon atoms; a radical -(CH2)n'-N3; or a radical -(CH2)n'-NR,8R,9 in which R18 represents a group removable by acid hydrolysis or by hydrogenolysis or an alkyl radical having from 1 to 4 carbon atoms and R19 represents a hydrogen atom or 5 an alkyl radical having from 1 to 4 carbon atoms, or R18 and R19 together with the intervening nitrogen atom form a phthalimido group.

2. A compound according to claim 1 of the general formula:

NHR16 5"-N --ú "I- C02H N \11 0 in which R16, Wand R5a are as defined in claim 1.

3. A compound according to claim 2 of the general formula:

(X111) "II(CH2)nl-R5a NIAR 1 16 M[V) 25 C02H N 30 0 \ (CH2Wi - Hcit in which W16 represents a trityl or chloroacetyl radical, n'i represents 1 or 2 and Hal' represents a bromine or iodine atom.

4. A compound according to claim 1, wherein R5,, represents a bromine or iodine atom or a phenylthio, 2-pyridinyithio,2-amino-1,3,4-thiadiazol-5-yithio,l-methyl-1H-tetrazol-5yit hio,2-amino-phenyithio,5-nitro- 40 2-pyridinylthio or 3-cyano-6-methyl-2-pyridinyithio group.

5. A compound according to claim 1, wherein R"i is a radical -CO-R' and R' represents an alkyl radical having from 2 to 4 carbon atoms, a phenyl radical or a radical -(CH2),- NR"2R"3 in which n is 0 or 1.

6. A compound according to claim 1, wherein R"i is a radical W4 1 -CH-COOK' and W4 represents a phenyl radical.

7. A compound according to claim 1, wherein Wi is they lactone of the formula 55 0

8. A compound according to claim 1, wherein Wi is a radical -(CH2)n'-R'5 in which n'is 1 or 2 and W5 60 represents an acetyl radical.

9. A compound as claimed in claim 1, wherein R"i represents a benzoyl, phthalimidoacetyl, phthalimidomethyl or tetrazol-5-yl-methyl radical.

46 GB 2 101 117 A

10. Acompound according to claim 1 of the general formula:

46 NHIR 16 (V11 L) 5 ". N 5 \=-, C02H N 10 0 1\ R b in which R16 and Rb are as defined in claim 1.

11. A compound according to anyone of claims 1, 2 or4to 10, wherein R16 is a formyl, acetyl, ethoxy--arbonyi, mesyl trifluoroacetyl, chloroacetyl, trityl, tert-pentoxycarbonyl, tert-butoxycarbonyl or benzyloxy carbonyl radical.

12. A compound according to claim 1, wherein R16 and/or Wi are substantially as hereinbefore described specifically.

13. A functional derivative of a compound according to anyone of the preceding claims which can react with a compound of the general formula ([X) as hereinbefore defined.

14. A functional derivative according to claim 13 which is a halide, asymmetric or mixed anhydride, an amide or an activated ester.

15. A compound according to claim 1 or a functional derivative thereof substantially as hereinbefore 25 described in any one of the specific Examples.

16. A process for preparing a compound of general formula VIA, as hereinbefore defined, which process comprises treating a compound of the general formula:

NHR16 (V) 30 N \-- C02H 35 N OH 40, wherein R16 is as defined in claim 1, with a functional derivative of the acid HO-CX-R'i3, wherein X and Wi3 40 are as defined in claim 1.

17. A process for preparing a compound of general formula WA as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the formula X=C=NH, wherein X is as defined in claim 1.

18. A process for preparing a compound of general formula V113 as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of general formula Y-CHW4-CO2A- in which Y represents a halogen atom or a sulphate or sulphonate group and W4 and A- are as defined in claim 1.

19. A process for preparing a compound of general formula Vie as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the general 50 formula Y \1 /,/' 55 wherein Y is as defined in claim 18.

20. A process for preparing a compound of general formula VID as hereinbefore defined, which process comprises reacting a compound of the general formula V as defined in claim 16 with a compound of the 60 general formula Y-(C1-12),-W5, wherein Y is as defined in claim 18 and n'and W5 are as defined in claim 1.

21. A process for preparing a compound of general formula VIE as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the general formula Y-(C1-12)12-CN, wherein Y is as defined in claim 18 and n2 is as defined in claim 1.

47 GB 2 101 117 A 47

22. A process for preparing a compound of general formula VIF as herein before defined, which process comprises treating a compound of the general formula:

NHR16 (V11) 5 ". N 5 C02H N 10 1% 0 - (CH2W -C N wherein R16 and n' are as defined in claim 1 either with hydrogen sulphide or, when n' as an integer other than 1, by hydrolysis in the presence of a base.

23. A process for preparing a compound of general formula VIG as hereinbefore defined, which process comprises treating a compound of general formula VIF as hereinbefore defined, wherein X' represents a sulphur atom, with a compound of the general formula W7-CO-CH2-Hal, wherein W7 is as defined in claim 1 and Hal is halogen.

24. A process for preparing a compound of general formula ViH as hereinbefore defined, which process 20 comprises treating a compound of general formula V11 defined in claim 22 with an azide.

25. A process for preparing a compound of general formula WH as hereinbefore defined, which process comprises treating a compound of the general formula:

NHR16 -1N C02Re N 11 OH wherein % represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, with a compound of the general formula Hal-(CH2),-Hal, wherein the substituents Hal, which are halogen, may be the same or different and n' is as defined in claim 1, to obtain a compound of the general formula:

(V1 25 N H R 16 (V]/W 40 C02Re 45 N 0 (CH2)ni- Hat 50 which, when % represents a hydrogen atom, is a compound VN and, when R, represents an alkyl radical having from 1 to 4 carbon atoms, is treated first with a base and then with an acid to yield a compound VN. 55

26. A process for preparing a compound of general formula Vij as herein before defined, which process 55 comprises reacting a compound of general formula VN as hereinbefore defined with pyridine.

27. A process for preparing a compound of general formula VIK as hereinbefore defined, which process comprises reacting a compound of general formua VN as hereinbefore defined with an azide.

28. A process for preparing a compound of general formula VIL as herein before defined, which process comprises reacting a compound of general formula VN as hereinbefore defined with an amine of general 60 formula MR1BR19, wherein R18 and R19 are as defined in claim 1.

29. A process for preparing a compound of general formula VI'Las hereinbefore defined, which process comprises reacting a compound of general formula VN as hereinbefore defined with imidazole, morpholine or an Walkyl piperazine.

30. A process for preparing a compound of general formula Vim as hereinbefore defined, which process 65 48 GB 2 101 117 A 48 comprises reacting a compound of general formula VN as hereinbefore defined with a compound of general formula Rar-SH, wherein Ra, is as defined in claim 1.

31. A process for preparing a compound of general formula (B) defined in claim 1 substantially as 5 hereinbefore described.

Printed for Her Majesty's Stationery Office, by Croydon Printing Company Limited, Croydon, Surrey, 1983. Published by The Patent Office, 25 Southampton Buildings, London, WC2A lAY, from which copies may be obtained.

31. A process for preparing a compound of general formula (B) defined in claim 1 substantially as hereinbefore described.

New claims or amendments to claims filed on 28 June 1982 Superseded claims 1-31 New or amended claims:- 1. A compound of the general formula:

NHR16 5"-- N -" C02H N _1 (B) ORi wherein R16 represents a protecting group for the amino radical and Wi represents:

a radical -CX-Wi3 in which Wi3 represents an alkyl radical having from 2 to 4 carbon atoms, an alkoxy group having from 1 to 4 carbon atoms, a phenyl radical or a radical -(CH2),,- NR"2R"3, in which n represents 0 oran 25 integer of from 1 to 4 and W2 and W3, which may be the same or different, each represent a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms with the proviso that both W2 and W3 do not both represent hydrogen when n is zero, or W2 and W3 together with the intervening nitrogen atom represent a phthalimido, piperidino or morpholino group and X is oxygen or sulphur; a radical -CX-NH2; a radical -CHR'4-C02A"', in which W4 represents a phenyl or cyano radical and Arepresents an easily-cleavable ester group, a radical -1 0 a radical -(CH2)r,,-R'5, in which n' is an integer of from 1 to 4 and W5 represents an alkoxy radical having 40 from 1 to 4 carbon atoms, a radical Alk-SQ)nsin which Alk represents an alkyl radical containing from 1 to 4 carbon atoms and ns is 0, 1 or 2, or an acyl radical having from 2 to 4 carbon atoms; a radical -(CH2),2-CN in which n2 is 2, 3 or 4; a radical -(CHAW-CX'- N112, in which X' represents a sulphur atom or, when n' is an integer otherthan 1, an oxygen atom, a radical in which W7 represents a methyl or amino radical; a radical a radical N CC - N -N H a - (CM2.)r,i 3/ a radical -(CH2)n-R5a in which R51 is a halogen atom, or a radical -S-Rar in which Rar represents a phenyl radical or a 5- or 6-membered aromatic heterocyclic radical containing from 1 to 4 heteroatoms selected from sulphur, nitrogen and oxygen, the phenyl and heterocyclic radicals being optionally substituted by one 65 49 GB 2 101 117 A 49 or more radicals selected from amino, nitro and cyano radicals and alkyl radicals having from 1 to 4 carbon atoms; a radical Rb which is a radical -(CH2)n'-R5b in which R5b represents an imidazolyl, morpholinyl or Nalkyl piperazinyl radical, the Walkyl radical containing from 1 to 4 carbon atoms; a radical -(CH2)n'-N3; or a radical -(CH2)n'-NR,8R,gin which R18 represents a g rou p removable by acid hydrolysis or by hydrogenolysis or an alkyl radical having from 1 to 4 carbon atoms and R19 represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, or R18 and R19 together with the intervening nitrogen atom form a phthalimido group.

2. A compound according to claim 1 of the general formula: 10 NHR16 (X111) "-- N \-- C02H 15 N \II 0 'I-I(CH2)n'-R5c, 20 in which R16, n' and R5a are as defined in claim 1.

3. A compound according to claim 2 of the general formula:

NIHIR 1 N 16 17--C02H N 0 (Xiill) (CH2W, - HcLL in which W16 represents a trityl or chloroacetyl radical, n'i represents 1 or 2 and Hal' represents a bromine or 40 iodine atom.

4. A compound according to claim 1, wherein R5a represents a bromine or iodine atom or a phenylthio, 2-pyridinyithio,2-amino-1,3,4-thiadiazol-5yithio,l-methyi-1H-tetrazol-5-yit hio,2-amino-phenyithio,5-nitro2pyridinylthio or 3-cyano-6-methyl-2-pyridinyithio group.

5. A compound according to claim 1, wherein Wi is a radical -CO-R' and R' represents an alky radical having from 2to 4 carbon atoms, a phenyl radical or a radical -(CH2)n- NR"2R"3 in which n is 0 or 1. 45 6. A compound according to claim 1, wherein R"i is a radical W4 1 -CH-COOK' and W4 represents a phenyl radical.

7. A compound according to claim 1, wherein R"i is they lactone of the formula -1 () 0 8. A compound according to claim 1, wherein R"i is a radical -(CH2)n-W5 in which n'is 1 or2 and W5 60 represents an acetyl radical.

9. A compound as claimed in claim 1, wherein R"i represents a benzoyi, phthalimicloacetyl, phthalimi domethyl or tetrazol-5-yl -methyl radical.

GB 2 101 117 A 10. A compound according to claim 1 of the general formula:

NHR16 5 N "" C02H N \ 0 (VI 1 L) 10 \ P b in which R16 and Rb are as defined in claim 1.

11. A compound according to anyone of claims 1, 2 or4to 10, wherein R16 is a formyl, acetyl, ethoxycarbonyi, mesyi, trifluoroacetyl, chloroacety], trityl, tert-pentoxycarbony], tert-butoxycarbonyl or benzyloxy carbonyl radical.

12. A compound according to claim 1, wherein R16 and/or Wi are substantially as hereinbefore described specifically.

13. A functional derivative of a compound according to anyone of the preceding claims which can react 20 with a compound of the general formula (R) as hereinbefore defined.

14. A functional derivative according to claim 13 which is a halide, asymmetric or mixed anhydride, an amide or an activated ester.

15. A compound according to claim 1 or a functional derivative thereof substantially as hereinbefore described in any one of the specific Examples.

16. A process for preparing a compound of general formula VIA as hereinbefore defined, which process comprises treating a compound of the general formula:

NHR16 N \-- "I- C02H N (V) OH wherein R16 is as defined in claim 1, with a functional derivative of the acid HO-CX-Wi& wherein X and Wi3 40 are as defined in claim 1.

17. A process for preparing a compound of general formula WA as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the formula X=C=NI-1, wherein X is as defined in claim 1.

18. A process for preparing a compound of general formula VIB as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of general formula Y-CHW4-CO2A- in which Y represents a halogen atom or an -S04H or an organic sulphonyloxy group and W4 and A- are as defined in claim 1.

19. A process for preparing a compound of general formula Vic as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the general 50 formula , h 0 wherein Y is as defined in claim 18.

20. A process for preparing a compound of general formula VID as hereinbefore defined, which process comprises reacting a compound of the general formula V as defined in claim 16 with a compound of the 60 general formula Y-(C1-12),-W5, wherein Y is as defined in claim 18 and n'and W5 are as defined in claim 1.

21. A process for preparing a compound of general formula VIE as hereinbefore defined, which process comprises reacting a compound of general formula V as defined in claim 16 with a compound of the general formula Y-(CHAn2-CN, wherein Y is as defined in claim 18 and n2 is as defined in claim 1.

51 GB 2 101 117 A 51 22. A process for preparing a corn pound of general form u la VIF as herein before defined, which process comprises treating a compound of the general formula:

NHR16 (V11) 5 ". N 5 \-- "I- C02H N C) -(CH2W -C N wherein R16 and n' areas defined in claim 1 eitherwith hydrogen sulphide or, when n' isan integerother 15 than 1, by hydrolysis in the presence of a base.

23. A process for preparing a compound of general formula VIG as hereinbefore defined, which process comprises treating a compound of general formula VIF as hereinbefore defined, wherein X' represents a sulphur atom, with a compound of the general formula W7-CO-CH2-Hal, wherein W7 is as defined in claim 1 and Hal is halogen.

24. A process for preparing a compound of general formula VIH as hereinbefore defined, which process 20 comprises treating a compound of general formula Vil defined in claim 22 with an azicle.

25. A process for preparing a compound of general formula VN as. hereinbefore defined, which process comprises treating a compound of the general formula:

NHR16 (T) 25 N C02Re 30 N OH 35, wherein Re represents a hydrogen atom or an alkyl radical having from 1 to 4 carbon atoms, with a 35 compound of the general formula Hal-(CH2L-Hal, wherein the substituents Hal, which are halogen, may be the same or different and n' is as defined in claim 1, to obtain a compound of the general formula:

N H R 16 (VI"H) 40 C 02R 45 N 0 '1-1 (CH2) ni - HaL 50 which,when Re represents a hydrogen atom, is a compound VN and, when % represents an alkyl radical having from 1 to 4 carbon atoms, is treated first with a base and then with an acid to yield a compound WH. 55 26. A process for preparing a compound of general formula VIj as herein before defined, which process 55 comprises reacting a compound of general formula WH as hereinbefore defined with pyridine. 27. A process for preparing a compound of general formula VIK as hereinbefore defined, which process comprises reacting a compound of general formula VN as hereinbefore defined with an azide. 28. A process for preparing a compound of general formula VIL as hereinbefore defined, which process comprises reacting a compound of general formula VN as hereinbefore defined with an amine of general 60 formula NHIR18R19, wherein R18 and R19 are as defined in claim 1.

29. A process for preparing a compound of general formula VI'Las hereinbefore defined, which process comprises reacting a compound of general formula VN as hereinbefore defined with imidazole, morpholine or an Walkyl piperazine.

52 GB 2 101 117 A 52 30. A process for preparing a compound of general formula Vim as hereinbefore defined, which process comprises reacting a compound of general formula WH as hereinbefore defined with a compound of general formula Rai--SH, wherein Rar is as defined in claim 1.