CA1072541A - PROCESS FOR PREPARING 7.beta.-ACYLAMINO-7-ALKOXYCEPHALOSPORINS OR 6.beta.-ACYLAMINO-6-ALKOXYPENICILLINS - Google Patents

Abstract

A B S T R A C T
.

7.beta. -Acylamino-7.alpha. -alkoxycephalosporins and 6.beta. -acylamino-6.alpha. -alkoxypenicillins are pre-pared by reacting a dialkoxyimino penicillin or cephalosporin compound with a halo-silane or an acid, and with water.

Description

)t7~5~1, The presen-t invention relates to a process for the~
preparation of derivatives of the ~-lactam antibiotics i.e. the cephalosporins and penicillins, and specifically to the prepar-ation of 7~-acylamino-7~-alkoxycephalosporins 63-acylamino-6~-alkoxypenieillins and certain intermediates in said process.
The cephalosporin derivatives referred to in the present specifieation are named with referenee to cepham, whieh has the strueture:

I 7 ~lS ~
~ ~ :

as described in J. Amer. Chem. Soc. 84, 3400 (1962), the corresponding unsaturated compounds being referred to as "cephems" and the position of the unsaturation being indicated by a numeral, e.g. 3-cephem.
The penicillin derivatives referred -to in the present specification are named as derivatives of penicillanic acid, whieh has the formula:

/ 1 \ / CH3 C~3 o COOH

., ~

.?,Y ~

Many cephalosporin and penicillin derivatives having antibiotic properties are known, the first of these to be discovered having various substituted acetamido chains at the 7~
or 6~ positions and being unsubstituted at the 7~ or 6cl positions, respectively. More recently, however, it has been discovered that various 7c~-or 6~-alkoxy derivatives of these compounds are also valuable antibiotic substances.
Of the known methods of introducing an alkoxy group into the 7- position of a cephem ring or the 6- position of a 10 penam ring, alkoxylation with t-butyl hypochlorite and lithium alkoxide is simplest to perform and gives best yields [R.A.
Firestone and B.G. Christensen, J. Org. Chem. 38, 1436 (1973);
G.A. Koppel and R.E. Koehler, J. Amer. Chem. Soc. 95, 2403(1973)];
however, this method has the disadvantage that it is not applic-able to cephalosporins or penicillings which are sensitive to t-butyl hypochlorite, i.e. which have an anion formation centre in the side chain at the 7- or 6- position.
We have now dïscovered a process for preparing certain 7~-acy]amino-7cc-alkoxycephalosporin or 6~-acylamino-6cl-20 alkoxypenicillin derivatives which are useful as antibacterialagents and some of which may also be used as intermediates in the preparation of other, but mor~ valuable, antibacterial agents.

: 30 .,

-2--The compounds which may be prepared by the process of the present invention have the formula (I):
Rl oR3 \ C.._C _ NH

R2 ~ ¦ ol ~ N Z (I) H ~ ~

wherein Rl represents a hydrogen atom, a halogen atom, an alkyl group having from 1 to 4 carbon atoms or an aryl group;
R2 represents a hydrogen atom, a halogen atom, an alkyl group having from 1 to 4 carbon atoms, and aryl group, an alkylthio group having from 1 to 4 carbon atoms, an alkynylthio group having from 2 to 4 carbon atoms, an aryl-thio group, an azidoalylthio group having from 1 to 4 carbon atoms, a cyano-alkylthio group having from 1 to 4 carbon atoms in the alkyl molety, an alkysulphonyl group having from 1 to 4 carbon atoms, a 5- or 6- membered heterocyclic group, a 5- or 6- membered heterocyclic-thio group, or 1 5- or 6- membered heterocyclic-oxy group;
R3represents an alkyl group having from 1 to 4 carbon : 20 atoms; and Z represents a fragment of formula:

.: R4 wherein R4 represents a carboxyl group, an alkoxycarbonyl yroup having from 1 to 4 carbon atoms in the alkyl moiety, a halo-alkoxycarbonyl group having from 1 to 4 carbon atoms in the alkyl moiety, a benzyloxycarbonyl group which is "' 5~

unsubstituted or has one or more halogen, methoxy or nitro substituents, a diphenylmethoxycarbonyl group, a triaakylsily-oxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety, a dialkylhalosilyloxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety a phenacyloxycarbonyl group which is unsubstituted or has one or more halogen or nitro sub-stituents, an acyloxycarbonyl group, a haloacyloxycarbonyl group, a dihalophosphinoxy-carbonyl group, a dialkylphosphinoxy-carbonyl group or an aminocarbonyl group; and A represents a hydrogen atom, an azido group or a group of formula - B-E wherein B represents an oxygen or a sulphur ~
atom and E represents an acyl group, an alkyl group having from ~ -1 to 4 carbon atoms or a substituted or unsubstituted carbamoyl, thiocarbamoyl or heterocyclic group.
In this formula (I), Rl is a hydrogen a~om; a halogen atom (e.g.chlorine , bromide or iodine); an alkyl group having from 1 to 4 carbon atoms (e.g. methyl, ethyl, n-propyl, iso-propyl or n-butyl); or an aryl group (e.g. phenyl or naphthyl).
R2 is preferably: a hydrogen atom; a halogen atom (e.g. chlorine, bromide or iodine); or an alkyl group ., ( --\
zs~
having from 1 to 4 carbon atoms (e.g. methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl or t-butyl~; and aryl group (e.g.
phenyl or naphthyl); an~alkylthio group having from 1 to 4 carbon atoms (e.g. methylthio, ethylthio, n-propylthio or iso-propylthio): an alkynylthio group having from 2 to 4 earbon atoms (e.g. propargylthio); an arylthio group (e.g. phenylthio);
an azidoalkylthio group having from 1 to 4 carbon atoms (e.g.
azidomethylthio or azidoethylthio); a eyanoalkylthio group having from 1 to 4 earbon atoms in the alkyl moiety (e.g.
cyanomethylthio or cyanoethylthio!; a 5- or 6- membered hetero-cyclic-thio group eontaining one or more sulphur and/or nitrogen and/or oxygen atoms in the ring and whieh may be substituted by an alkyl group havingfrom 1 to 3 earbon atoms (e.y. imidazolyl-- thio, thiadiazolylthio, triazolylthio, thienylthio, isoxazolyl-thio, methylisoxazolylthio, tetrazolylthio, methyl-tetrazolyl-thio, pyrimidinylthio or pyridylthio); a 5- or 6- membered heterocyclie-oxy group eontaining one or more sulphur and/or nitrogen and/or oxygen atoms in the ring and whieh may be sub-stituted by an alkyl group having from 1 to 3 earbon atoms (e.g.
isoxazolyloxy methylisoxazolyloxy, imidazolyloxy, thiadiazolyl-o~xy, triazolyloxy, thienyloxy, tetrazolyloxy, methyltetrazoly-loxy, pyrimidinyloxy, or pyridyloxy); a 5- ;

.

~ 30 .,.

~ 5-~

or 6- membered heterocyclic group containing one or more sulphur and/or nitrogen and/or oxygen atoms in the ring and which may be substituted by an alkyl group having from 1 to 3 carbon atoms (e.g. thienyl, imidazol, thiadiazolyl, isoxazolyl, methylisoxazolyl, tetrazolyl, methyl-tetrazolyl, pyrimidinyl or pyridyl); or an alkylsulphonyl group having from 1 to 4 carbon atoms (e.g. methylsulphonyl, ethylsulphonyl, or n-propylsulph-onyl). R is an alkyl group having from 1 to 4 carbon atoms (e.g. methyl, ethyl, n-propyl, isopropyl or n-butyl). R4 is a carboxyl group or a protected carboxyl group such as an alkoxycarbonyl group having from 1 to 4 carbon atoms in the alkyl moiety (e.g. methoxycarbonyl, ethoxycarbonyl, n-propoxy-carbonyl or n-butoxycarbonyl), a haloalkoxycarbonyl group having from 1 to 4 carbon atoms in the alkyl moiety (e.g.
dichloroethoxycarbonyl or trichloroethoxycarbonyl), a benzyl-oxycarbonyl group optionally substituted with halogen, methoxy or nitro (e.g. benzyloxycarbonyl, _-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl or p-nitrobenzyloxycarbonyl), a diphenylmethyloxycarbonyl group, a trialkylsilyloxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety (e.g.
trimethylsllyloxycarbonyl or triethylsilyloxycarbonyl), a dialkylhalosilyloxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety (e.g. dimethylchlorosi]yloxycarbonyl or dimethylbromosilyloxycarbonyl), a phenacyloxycarbonyl group optionally substituted with halogen or me-thoxy (e.g.
.

~-chlorophenacyloxycarbonyl, _ bromophenacyloxycarbonyl or _-methoxyphenacyloxycarbonyl), an acyloxycarbonyl group (e.g.
acetoxycarbonyl or benzoyloxycarbonyl), a halo-acyloxycarbonyl group (e.g. chloroacetoxycarbonyl or bromo-acetoxycarbonyl), a dihalophosphinoxycarbonyl group (e.g. dichlorophosphinoxycarbonyl or dibromophosphinoxycarbonyl), a dialkylphosphinoxy-carbonyl group (e.g. dimethylphosphinoxycarbonyl), or an aminocarbonyl group (e.g. 3-oxo-2, 3-dihydro-s-triazolo[4,3-a]pyridon-3-ylcarbonyl). E is preferably: an acyl group (e.g. acetyl, propionyl or benzoyl); a carbamoyl group; or a 5- or 6- membered heteroxyclic group, which may be substituted by an alkyl group having from 1 to 3 carbon atoms (e.g. tetrazolyl, l-methyltetra-zolyl, isoxazolyl, imidazolyl, thiazolyl, triazolyl, thienyl, thiadiazolyl, methylthiadiazolyl, pyrimidinyl or pyridyl).
In accordance with the invention which is the subject of our application No. 222728, compounds of formula (I) are prepared by reacting a halo-imine of formula (II):

:
Rl H

Y Y

(in which Rl, R2 and Z are as defined above and yl and Y are the same or different and each represents a halogen atom) with an alkali metal alkoxide of formula (I~

R OM (III~ ;

(in which R3 is as defined above and M represents an alkali metal) to give an alkoxyketeneimine of formula (IV):

Rl oR3 / C = C = N

0 (IV) (in which R , R and Z are as defined above) and/or a dialkoxy-imino compound of formula (V):

~` ' ' :

' .

: ~

~ 5~1 I
Rl OR

R ~1 OR ~ (V) O ~ ,,, (in which Rl, R2, R3 and Z are as defined above), and hydrolizing said alkoxyketeneimine (IV) and/or reacting said dialkoxyimino --compound (V) with a halo-silane or an acid and treating the resulting product with water~
The process which is the subject of the present invention is the preparation of a compound of formula (I) by the reaction of a dialkoxyimino compound (V) with a halo-silane or an acid and treatment of the resulting product with water. The alternative route via an alkoxyketeneimine of formula (IV) is ; the subject of our copending application 321,402 of even date.
The first stage in the reaction of the halo-imine (II) with the alkali metal alkoxide (III) to form the alkoxyketene-imine (IV) and/or the dialkoxyimino compound (V) is the dehydro-halogenation of halo-imine (II). This reaction must, thereore, be carried out in the presence of an acid-binding agent, i.e.
a base. If desired, the base may be provided by the alkali metal alkoxide (III) or, alternatively, another base may be used. Preferably, the base is employed in an amount of at least 1 equivalent per mole of halo-imine (II) and more preferably about 1 equivalent is employed. The course of the subsequent reaction depends upon the relative proportions of alkali metal alkoxide and haloimine (II). Where the amount of alkali metal 30 alkoxide (excluding that, if any, employed as acid-binding agent in the preceding stage) is about 1 mole per mole of halo-imine , , -9-", :'1,~3~,t (II~, the product of the reaction is predominantly the alkoxy-keteneimine (IV). On the other hand~ if the amount of alkali metal alkoxide is about 2 mole per mole of halo-imine (II), the dialkoxyimino compound (V) is -the predominant product, as re-quired for use in the process of the present invention. Where the amount of alkali metal alkoxide is between 1 and 2 moles per mole of halo-imine (II), a mixture of alkoxyketeneimine (IV) and dialkoxyimino compound (V) will be obtained. Although the presence of excess alkoxide will have no adverse effect, it is not beneficial and, accordingly, we prefer to employ 2 moles of alkoxide per mole of halo-imine (II), in order to produce predominantly dialkoxyimino compound (V). (About 1 mole of alkoxide per mole of halo-imine (II) will produce predominantly alkoxyketeneimine (IV)) dialkoxyimino compound (V).

. .

5~:~

The halo-imines of formula (~I~ are themselves new compounds. ~hey may be prepared by reacting an acylamino compound of formula(VI):

Rl H

C - C - N~ ¦ ~

X N ~ tVI) (in which Rl, R and Z are as defined above and X represents a hydroxy group or a halogen atom) with a halogenating agent.
The reactions of which ihe process of the present invention forms part may be illustrated by the following reaction scheme:

, ., .
,. .

; 30 ~' ~"

:1~'7~5~1 1 R\ H S
C C NH --/ \
RV'¦ II ¦ ~ halogenate ~, X O ~ J~ Z
0~ ~J :
(VI ) Rl H
~II) \C _ C =N

Y Y O
~--Rl _ ¦ - .

~_ \ C = C_ N

Rl (VI I ) OR S
R30 M ~, f ~ C=N~
R
(IV) ~"~or~

Rl OR

RIl ~R3 , (V) / ha los 1 lane : ;/ H 2 OR ?
\C__C NH 5 ~S~
R H O O~ ~ z (I) : .

' ' 5~1 Thus, in the first step o~ the overall process forming the subjec~ of our said application No. 222728, halo-imine (II) is prepared by reacting acylamino compound(VI) with a halogenating agent. Examples oE halogenating agents are: phosphorus pent-ahalides, e.g. phosphorus pentachloride or phosphorus penta-bromide; phosphorus trihalides, e.g. phosphorus trichloride or phosphorus tribromide; phosphorus oxyhalides, e.g. phosphorus oxychloride or phosphorus oxybromide; and thionyl halides, e.g.
thionyl chloride or thionyl bromide. Of these, phosphorus pentachloride is most preferred. The halogenta-tion reaction is ` suitably carried out in an anhydrous inert organic solvent, with stirring, at a temperature from -50C to 40C. The reaction time is not critical, but the reaction will normally be complete `-~ within a period from 15 minutes tc 5 hours. Suitable inert organic solvents are chloroform, methylene chloride, benzene, diethyl ether, tetrahydrofuran and dioxane. The reac-tion is - preferably effected in the presence of a tertiary amine, such as quinoline, diethylaniline, dimethylaniline, pyridine, triethyl-amine or diazabicyclooctane.
When the halogenation reaction is complete, the reaction mixture, including compound (II), may be used as such as the starting material in the next s-tep; alternatively, the compound(II) may be recovered and purified by conventional means.

!
. .

';
'"

; -l3-5~

~ here R in the acylamino starting material (VI) is a carboxyl group, it should first be pro~ected, by methods which are well understood in the art. If X in the starting material represents a hydroxy group, the halogenation reaction causes it to be replaced by a halogen atom.
Dialkoxyimino compound (V) is prepared by reacting halo-imine (II) with two moles of alkoxide (III) in the presence of one equivalent of a base, per mole of halo-imine, in an inert organic solvent. Examples of alkali metal alkoxides include:
lithium methoxide, lithium ethoxide, sodium methoxide, sodium ethoxide, potassium methoxide and potassium ethoxide. The base employed may be, and preferably is, simply an excess of the alkali metal alkoxide (III). Alternatively, it may be: an inorganic 2~

'~

.

:. .
'.

', Jz~

base, such as an alka]i metal hydroxide (e. g. sodium hydroxide or potassium hydro~ide), an alkali metal carbonate (e. g. sodium carbonate or potassium carbonatej, or an alkali metal hydride (e. g, sodil1m hydride or potassium hydride); or a tertiary amine, such as a tria]kylamine (e, g. trimethylamine or triethylamilie), a dialkylaniline (e. g. dimethylaniline or diethylaniline), pyridine, quinoline or diazabicyclooctane, The reaction is prefera~ly carried - out in an inert organic solvent at temperatures of from -7~C to 20C and will normally require a time of from 1 to 60 minutes.
Suitable inert organic solvents are chloroform, diethyl ether, tetrahydrofuran, dioxane, benzene and alcohols of formual ~ OH in which R is as defined above. After the reaction is cornplete, the reaction mixture can be used as such as the starting material for the next step in the process. Alternatively, the product may, if desired, be recovered and purified by conventional means, Compound (I) can then be obtained by reacting dialkoxyimino compound (V) with a halo-silane or with an acid and treating the resulting product with water. Examples of halo-silane which may be employed in this reaction include: trialkylhalosilanes, such as trimethylchlorosilane, trimethylbromosilane, triethylchlorpsilane and triethylbromosilane;
and dialkyldihalosilanes, such as dimethyldichlorosilarle, dimethyl-dibromosilane, diethyldichlorosilane and diethyldibromosilane. Examples of acids which may be used include: mineral acids, such as hydrochloric acid, hydrobron1ic acid, sulphuric acid, phosphoric acid and perchloric ~' acid; and Lewis acids, such as boron trifluoride, boron trichloride, aluminium trichloride, tin ~IV)chloride or titanium (IV) chloride. The reaction is preferably carried out by dissolving dialkoxyimino compound (V) in an inert organic solvent (e.g. benzene, toluene, dioxane, diethyl ether, carbon tetrachloride, chloroform or me-thylene chloride), adding the halo-silane or the acid to the solution, stirring the mixture at a temperature from -50C to 40C for the required reaction time (generally from 5 minutes to 24 hours) and adding the resulting mixture to water. If the mixture produces a strongly acidic solution, it is desirable to add it to a cold buffer solution (pH4 - 8).
After completion of the reaction, the desired product may be recovered by conventional means; for example, the reaction mixture is extracted with an appropriate organic solvent, the extract is washed with water and dried and the solvent is then evaporated off. The residue may be purified ~ ~ .
- by recrystallation or chromatography.
(Instead of proceeding via dialkoxyimino compound (V), the overall process may proceed via alkoxyketeneimine (IV) in accordance with the invention of our said copending application ; 321,402 of even date).
Some of the acylamino compound (VI) employed as starting materials in the overall process are novel and may be ; prepared by known methods, for example those described in "Cephalosporins and Penicillins" edited ',', :
. .

:, ., ''.,' '"'.
' by E, H, Flynn, ~cademic Press, New York and London (1972) page 83. By way of example, they may be prepared by reacting the corresponding amino compound with a halide or anhydride of a carboxylic ac:id having the formula:

R
C - C 00~1 .. ..

5~

(in which Rl~ R2 and X are as defined above) or by reactiny the corresponding amino compound with the earboxylic acid itself in the presence of a peptide coupling agent e.g. dicyclohexylurea.
The process of the present invention has the advantag-es that is generally applicable to the preparation of cephalo-sporins and penieillins, even if they have an anion formation eentre in the side chain, and that it ean be carried out as part of a sequence without isolation of the intermediates produc-ed during the reactions, i.e. in a "one-pot" reaction.
Certain of the compounds of formula(I) are new.
Examples of the new compounds are the following:
(1? 7~-chloroacetamido-7~-methoxy-3-methyl-3-eephem-4-earboxylie acid.
(2) 3-earbamoyloxymethyl-7~-ehloroacetamido-7~-meth-oxy-3-cephem-4 earboxylie aeid.

(3) 7~-methoxy-3-methyl-7~-propionamido-3-eephem-4-earboxylie acid.

(4) 7~-methoxy-3-methyl-7~-phenoxyacetamido-3-cephem-4-earboxylie acid.

(5) 3-aeetoxymethyl-7~-methoxy-7~-phenylthioaeetamido-3-eephem-4-earboxylic aeid.

(6) 3-aeetoxymethyl-7~-ehloroaeetamido-7~-methoxy-3-eephem-4-earboxylie aeid.

~;, ' 30 ~.' :`
., .

Z'j~

(7) 7~-chloroacetamido-7~-methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4--carboxylic acid.

(8) 3-carbamoyloxymethyl-7~-methoxy-7~-(1,2,4-triazol-4H-3-yl)thioacetamido-3-cephem-4-carboxylic acid.

(9) 3-carbamoyloxymethyl-7~-(imidazol-2-yl)thioaceta-mido-7~-methoxy-3-cephem-4-carboxylic acid~

(10) 3-carbamoyloxymethyl-7~-methoxy-72-(1,3,4-thiadiazol-2-yl)thioacetamido-3-cephem-4-carboxylic acid.

(11) 3-carbamoyloxymethyl-7N-methoxy-7~-propargyl-thioacetamido-3-cephem-4-carboxylic acid.

(12) 3-carbamoyloxymethyl-7~-cyanomethylthioace-tamido-7~-methoxy-3-cephem-4-carboxylic acid.

(13) 3-carbamoyloxymethyl-7a-methoxy-7~-(5-methyl-1,2,4-triazol-4H-3-yl)thioacetamido-3-cephem-4-~arboxylic acid.

(14) 3-carbamoyloxymethyl-7~-methoxy-7~-(2-pyridyl)-thioacetamido-3-cephem-4-carboxylic acid.

(15) 3-carbamoyloxymethyl-7~-methoxy-7~-(2-pyrimidyl)-thioacetamido-3-cephem-4-carboxylic acid.

(16) 3-carbamoyloxymethyl-7~-methoxy-7~-(thiazolin-2-yl)-thioacetamido-3-cephem-4-carboxylic acid.

(17) 3-carbamoyloxymethyl-7~-methoxy-i3-(n-propyl-thioacetamido)-3-cephem-4~carboxylic acid.

' ' 30 ., ,' ' ., ,' ,. .

(18) 3-acetoxyme-thyl-7~-cyanomethylthioacetamido-7~-methoxy-3-cephem-4-carboxylic acid.
(l9) 3-acetoxymethyl-73-(imidazol-2-yl)thioacetamido-7~-methoxy-3-cephem-4-carboxylic acid.
(20) 7 ~methoxy-3 (1-methyl-lII-tetrazol-5-yl)thio-methyl-7~-(1,3,4-thiadiazol-2-yl)thioacetamido-3-cephem-4-carboxylic acid.
(21) 7~-(imidazol-2-yl)thioacetamido-7~-me-thoxy-3-(l-methyl-lH-tetrazol-5~yl)thiomethyl-3-cephem-4-carboxylic acid.
(22) 7~-cyanomethylthioacetamido-7~-methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.
(23) 7a-methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiometh-yl-7~-propargylthioacetamido-3-cephem-4-carboxylic acid.
(24) 1~-bromoacetamido-3-carbamoyloxymethyl-7~-methoxy-3-cephem-4-carboxylic acid.
(25) 6~-chloroacetamido-6~-methoxypenicillanic acid.
(26) 6~-methoxy-6~-phenylthioacetamidopenicillanic acid.
(27) 6~-methoxy-6~-propargylthioacetamidopenicillanic acid.
(28) 6~-methoxy-6~-(1,3,4-thiodiazol~2-yl)thioacetam-ido-penicillanic acid.

. .

'' ..

:, :

^J~tj~

(29) 7~-benzyloxy-7~-chloroace-tamido-3-methyl-3-cephem-4-carboxylic acid.
(30) 713-(isoxazol-3-yl)oxyace-tamido-7~-me-thoxy-3-(l-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.
(31) 7~-methoxy-3-(1-methyl-liI-tetrazol-5-yl)thio-methyl-7~-(5-methyl~1,3,4-thiodiazol-2-yl) thioacetamido-3-cephem-4-carboxylic acid.
(32) 3-carbamoyloxymethyl-7~-methoxy-73-(5-methyl-~, 3,4-thiadiazol-2-yl)thioacetamido-3-cephem-4-carboxylic acid.
(33) 3-acetoxymethyl-7~-azidomethylthioacetamido-7~-methoxy-3-cephem-4-carboxylic acid.
(34) 3-acetoxymethyl-7~-methoxy-7~-(1,3,4-thiadiazol-2-yl) thioacetamido-3-cephem-4-carboxylic acid.
(35) 3-acetoxymethyl-7~-methoxy-7~3-propargylthioacet-amido-3-cephem-4-carboxylic acid.
(36) 3-acetoxymethyl-7i3-(isoxazol-3-yl)oxyacetamido-7~-methoxy-3-cephem-4-carboxylic acid.

. . .
(37) 3-acetoxymethyl-7~-(isoxazol-3-yl)thioacetamido-7~-methoxy-3-cephem-4-carboxylic acid.
(38~ 7~-methoxy-73-methylsulphonylacetamido-3-(1-methyl-lH-tetrazol-5-yl)-thiomethyl-3-cephem-4-carboxylic acid.

1 , `~ 30 '' , O~ the compounds in the above list, the following.
numbered compounds have particularly good antimicrobial activity against a wide range of pathogenic microorganisms:
8; 9; 10; 11; 12; 18; 19; 20; 21; 22; 23; 30; 31; 32; 33; 34;
35; 36; 37; and 38. The antimicrobial activi-ties of these compounds, in terms of their minimum inhibitory concentrations (meg/ml) against various microorganisms, are summarized in the following Table in which the compounds are identified by the numbers given in the above list. The microorganisms against which the compounds were tested were as follows:
lA : _aphylococcus aureus 209 p IB : ~ hylococcus aureus .
(CP and PC resistant) IIA : E. coli NIHJ
IIB : E. coli 609 (CER resistant) III : Shigella flexneri Komagome - IV~ : Klebsiella pneumoniae 806 IVB : Klebsiella pneumoniae 846 (CER resistant) V : Proteus vulgaris VI : Salmonella entaritidis Gaertner : 30 2~4 ~L

Table ._ = Mlcroorganlsm Compound No. IA IBII~IIB III IVA ¦ IVB V ¦ VI
_ _~
8 1.5 6.26.212.512.56.2 >400 25 6.2 9 ~0.10.81.5 3.13.1 3.1 i200 6.2 1.5 0 0.4 1.5 3.13.1 3.13.1>400 6.2 1.5 0.8 1.5 3.13.1 6.23.1>400 6.2 1.5 2 0.8 1.5 1.50.8 3.10.4>400 6.2 0.4 8 0.2 0.8 1.51.5 3.11.5>400 6.2 0.8 9 1.5 6.212.5 25 2512.5>400 25 6.2 20 0.4 0.8 1.5 1.51.5 1.5>200 3.1 0.8 21 0.8 3.1 6.2 6.26.2 6.2>200 12.5 3.
22 0.2 0.8 0.8 0.80.8 0.8>200 1.5 0.2 23 0.2 0.8 3.1 3.13.1 3.1400 3.1 0.8 30 0.2 0.8 3.1 3.13.1 3.1>400 6.2 1.5 31 0,A 1.512.512.56.212.5400 3.1 1O5 32 0.8 3.1 6.2 6.212.56.2>200 6.2 3.1 L _ 4 ~8 6 26,2 6.26 2 >200 ~ 1~5 ;' ~ .
~';

.

r~

__ ~
Microorganism Compound ~ ~____ ~ _ No. IA ~ IIA IIB III IVA IVB ¦ V VI

340.8 1.5 3.1 6.26.2 3,1 ~400 6.2 3.1 350.8 3.1 3.1 6.26.2 6.2 >400 6.2 1.5 36<0.1 0.8 6.2 6.212.5 6.2 ~400 12.5 3.1 370.2 0.8 3.1 6.26.2 3.1 >400 3.1 1.5 ~ I' , ,'' .' .

' ' '~:
:

, . . .
." ' .

.
:`

;"
.,' ~

~ ~ J ~
Accordingly, the compounds produced by the process of the present invention have excellent antimicrobial activities againSt a wide ranye of pathogenic micro-organisms. The compounds may be administered orally or parenterally, for example as capsules, tablets or injections; they are most preferably administered by injection. The optimum dosage will vary with the body weight, age and disease of the patient but the total daily dosage will generally be from 100 to 3000 mg, administered in divided doses, three or four times a day; if necessary, however, more than this amount may be used.
In addition to their pharmaceutical use, the 7~-haloacetamidocephalosporin and 6~-haloacetamido-penicillanic acid derivatives, such as 7~-chloroacetamido-7 ~methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid and its esters, are useful intermediates, which can be trans-formed into other cephalosporin or penicillin derivatives having valuable antimicrobial activity by replacing the halogen atom with another radical.
The invention is illus-trated by the following Examples:

:~7 _XAMPLE I.
(A) Methyl 3-methyl-7~-1(1',2',2'-trichloroethylidenimino)-3-cephem-4-carboxylate 600 mg of phosphorus pentachloride were dissolved in 24 ml of chloroform containing 0.4 ml of quinoline. To the resulting mixture were then added 536 mg of methyl 7~-dichloro-acetamido-3-methyl-3-cephem-4-carboxylate. The reaction mixture was stirred for 1 hour and the chloroform was then removed by distillation in vacuo. To the residue were added 40 ml of 10 tetrahydrofuran and the resul-ting precipitates were filtered off.
The resulting tetrahydrofuran solution of methyl 3-methyl-7~-(1',2',2'-trichloroethylidenimino)-3-cephem-4-carboxylate was ~ used in the next state without purification.
- To isolate the pure compound, however, a tetrahydro-furan solution prepared exactly as described above was poured into a 10% w/v solution of dipotassium hydrogen phosphate and extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was . then evaporated off under reduced pressure at 20C, giving 530 ` 20 mg of methyl 3-methyl-7~-(1',2',2'-trichloroethylidenimino)-3-oephem-4-carboxy1ate, as an ~il.

.

.
I

'' NMR spectrum (CDC13) ~ ppm - 2.17 (3H, singlet), 3.27 & 3.50 (2H, AB
quartet, J = 19 Hz), 3.83 (3H, singlet), S.10 (lH, doublet, J = 5 H~
5.55 (lH, doublet, J = 5 Hz), 6.42 (lH, singlet.) (B) Preparation of Methyl 7~-(2'-chloro-1'-methoxyethyl-..
idenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylate ` A methanol solution of lithium methoxide prepared from 60 mg of lithium and 5 ml of methanol was added at -70C
to the tetrahydrofuran solution of methyl 3-methyl-73-(1',2',2'-trichlorethylidenimino)-3-cephem-4-carboxylate prepared above.
The reaction mixture was stirred at -70C for 30 minutes and then treated with 0.5 ml of acetic acid to decompose the excess li-thium methoxide. The solution was then poured into water and extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate, after which the solvent was evaporated off, giving 480 mg of me-thyl 7~(2'-chloro-1'-methoxyethylidenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylate, m.p. 138-139C.

~' ' i, .; .
' ~ 30 , .

~ ~3~

(C) Meth~rl 7~-chloroacetamido-7~-methoxy-3-methy--3--cephem-4-carboxylate 50 mg of the methyl 7~-(2'-chloro-l'-methoxyethyliden-imino)-7~-methoxy-3-methyl-3-cephem-4-carboxylate prepared above were dissolved in 4 ml of dry chloroform containing 0.05 ml of quinoline and 0.2 ml of trimethylchlorosilane. Nitrogen gas was then bubbled into this solution for 3 hours, after which the solution was poured into water and extracted with chloroform.
The extracts were washed with water and dried over anhydrous soldium sulphate, and then the solvent was evaporated off under - reduced pressure to give 45 mg of methyl 7~-chloro-acetamido-7u-methoxy-3-methyl-3-cephem-4-carboxylate, m.p. 123-125C.
IR spectrum (Nujol-Trade Mark) v maxcm = 3380, 1790, 1730, 1680, NMR spectrum (CDC13) ~ ppm = 2.25(3H, singlet), 3.20 (2H, singlet), 3.53 (3H, singlet), 3.73 (3H, singlet), 4.07 (2H, singlet), 4.98 (lH, singlet), 7.43 (lH, singlet).

.

~7~5~

Methyl 7~-chloroacetamido-7~-methoxy-3-methyl-3-cephem-4-carboxyl e 50 mg of methyl 7~-(2'-chloro-1~-methoxyethylidenimino) -7~-methoxy-3-methyl-3-cephem-4-carboxylate, prepared as describ-ed in Example l, were dissolved in 2 ml of dry tetrahydrofuran containing 0.1 ml of quinoline and 0.25 ml of boron trifluoride etherate. The reaction mixture was stirred overnight at room temperature and then poured into water and extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate and the solvent then evaporated off n vacuo. The residue was chromatographed on silica gel, giving 18 mg of methyl 7~-chloroacetamido-7~-methoxy-3-meth~1-3-cephem-4-carboxylate.

(A) p-Bromophenacyl 3-acetoxymethyl-7~-(1',2',2'-trichloro-eth lidenimino)-3-cephem-4-carboxvlate Y
A solution of p-bromophenacyl~3-acetoxymethyl-7~-(1', 2',2'-trichloroethylidenimino)-3-cephem-4-carboxylate in tetrahydrofuran was prepared from 290 mg of p bromophenacyl 3-acetoxy ethyl-73-.~ ~

A

.

t)~L

dichloroacetamido-3 cephem-4-carboxylate, 180 mg of phosphorus pentachloride and 0.12 ml of ~uinoline, according to the pro-cedure described in Example 1 (B) _-Bromophenacyl 3-acetoxymethyl-7~-(2'-chloro-1'-methoxy-ethylldenimino)-7~-methoxy-3-cephem-4-carboxylate A methanol solution of lithium methoxide prepared from 2] mg of lithium and 2 ml of methanol was added to the solution of p-bromophenacyl 3-acetoxymethyl-7~-(1'/2',2'-trichloroethyl-idenimino)-3-cephem-4-carboxylate in tetrahydrofuran prepared above, maintaining the temperature at -70C. The reaction mixture was stirred at -70~C for 30 minutes and then treated with 0.15 ml of acetic acid. The solution was poured into water and extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate, after which the solvent was evaporated off under reduced pressure giving 240 mg of p-bromophenacyl-3-acetoxymethyl-7~-(2'-chloro-l'methoxy-ethylidenimino)-7~-methoxy-3-cephem-4-carboxylate.
I~ spectrum (Nujol-Trade Mark) ~ cm 1 1780, 1740, 1700.

:`

`~ s~ :

NMR spectrum (CEC13) ~ ppm = 2.08 ~3H, singlet), 3.37 & 3.58 (2H, ~B
quartet, J = 19 Hz), 3.47 (3H, singlet)/
3.83 (3H, singlet), 4.17 & 4.50 (2H, AB
quartet, J = 13 Hz), ~ -5.03 (2H, AB quartet, ; ~-J = 13 Hz), 5.05 (lH, singlet), 5~43 & 5.57 (2H, AB
quartet 7 J = 16 Hz).
7.4-8.1 (4H, multiplet).
(C) ~-Bromophenacyl 3-acetoxymethyl-7~- chloroacetamido-7~-, methoxy-3-cephem-4 carboxylate 280 mg of the _-bromophenacyl 3-acetoxymethyl-7~-(2'- !
chloro-l'-methoxyethylidenimino)-7~ -methoxy-3-cephem-4-carboxylate prepared above were dissolved in 16ml of dry chloroform containing 0.2 ml of quinoline and 0.8 ml of trimethylchlorosilane. The reaction mixture was stirred at room temperature for 3 hours while nitrogen gas was introduced into the solution. The mixture was then pouxed into water and extracted with chloroform. The extracts were washed with a 10% w/v solution of sodium dihydrogen phosphate and with water, I and dried over magnesium .
:

sulphate. The solvent was then evaporated off under reduced J pressure, giving 25Q mg of p-bromophenacyl 3-acetoxymethyl-7~-chloxoacetamido-7~-methoxy-3-cephem-4-carboxylate.
NMR spectrum (CDC13) ~ ppm = 2.10 (3H, singlet~, 3.38 & 3.57 (2H, AB
quartet, J = 18 Hz), 3.58 (3H, singlet), 4.13 (2H, singlet), 5.05 & 5.13 (2H, AB
quartet, J = 14 Hz), 5.12 (lH, singlet), 5.50 (2H, singlet), 7.5-8.0 (4H).

(A) Benzhydryl 3-acetoxymethyl-7~-(1',2',2'-trichloroethyl-,' ---- ' .
idenimino)-3-cephem-4-carboxylate ':
A solution of this compound in tetrahydrofuran was prepared from 549 mg of benzhydryl 3-acetoxvmethyl-7~-dichloro-acetamido-3-cephem-4-carboxylate, 400 mg of phosphorus penta-chloride and 0.27 ml of quinoline, following the proceduredescribed in Example 1.

.
f ~ ' : ':
.

i::
~ 30 ' 5~ ~
I

(B? Benzhydryl 3-acetoxymethyl-7~-(2'-chloro-1'-methoxy-ethylidenimino)-7~-methoxy-3-cephem-4-carboxylate __ ~ _ _ A methanol solu-tion of lithium methoxide prepared from 40 mg of lithium and 4 ml of methanol was added to the solution of benzhydryl 3-ace-toxy-methyl-7~-(1',2',2'-trichloro-ethylidenimino)-3-cephem-4-carboxylate in tetrahydrofuran prepared above, whilst maintaining the temperature at -70C.
The reaction mixture was stirred at -70C for 30 minutes and then treated with 0.4 ml of acetic acid. The solution was poured into water and extracted with ethyl acetate. The combined extracts were washed with wa-ter and dried over anhydrous sodium sulphate; the solvent was evaporated off under reduced pressure, giving 480 mg of benzhydryl 3-acetoxymethyl-7~-(2'-chloro-l'-methoxyethylidenimino)-7~-methoxy-3-cephem-4- -carboxylate.
NMR spectrum (CDC13) ~ ppm = 2.05 (3H, singlet), 3.27 & 3.45 (2H, AB
quartet, J = 18 Hz), 3.42 (3H, singlet), 3.77 (3H, singlet), 4.10 & 4.45 (2H, AB
quartet, J = 16 Hz), 4.65 & 4.88 (21I, AB
quartet, J = 13 Hz), 4.94 (~H~ singlet), 6.85 (lH, singl~tj, 7.0 - 7.6 (lOH, multiplet)~

(C ~nzhy_ryl 3-acetoxymethy]-7~-chloroacetamido-7~- methoxy--3-cephem-4-ca _ ~ylate O~l ml of quinoline and 0.5 rnl of trimethyl-chlorosilane were added to a solution of 270 mg of benzhydryl 3-acetoxymethyl-7~-(2'-chloro-1'-methoxy-~; 10 ethylidenimino)-7~-methoxy-3-cephem-4-carboxylate in 12 ml of dry chloroform. Nitrogen gas was bubbled into this solution at room temperature for - 1 hour. A further 0.5 ml of trimethylchlorosilane was added and introduction of nitxogen gas was continued for a further 2 hours. The solution was then poured into water and extracted with chloroform.
The combined extracts were washed with water and dried over anhydrous sodium sulphate, after which the solvent was evaporated off. The residue was chromatographed on silica gel, giving 230 mg of - benzhydryl 3-acetoxymethyl-~7~-chloroacetamido-7~-~ Ame-thoxy-3-cephem 4-carboxylate.

,:
NMR spectrum (CDC13) ~ ppm ~ 1.97 ~3H, singlet), 3.30 & 3.~0 (2H, .~B

quartet, J ~ 17 H~), :

ZS9L~
.08 (3H.~sir~let) J
4.87 (2H, sing].et), - 5.02 & 5.07 (2H, AB
quar-tet, J ~ 14Hz), 5.04(1H, singlet), 6.95 (lH, s.inglet~, 7.2 - 7.6 ~lOH
multiplet), 0 Benzhydryl 3~ methyl-1H-tetrazol -5-yl)thiomethyl-7~-(1',2',2'-tri.chloroethylidenimino)-3-cephem~4-carboxylate ..... _ . _ " ~ A solution of this compound in tetrahydrofuran ~ was prepared from 314 mg of benzhydryl 7~- dichloroa-cetamido-3-(1-methyl-1H-tetrazol -5-yl)thiromethyl-~-cephem-4-carboxylate, 180 mg of phosphorus pentachloride and 0013 ml of quinoline, according to the procedure described in Example 1-....

. (B) Benzhydryl 7~-(2'-chloro-l'~methoxyethyl~deni-mino)-7~-methoxy-3~(1-methyJ.-lH-tetrazol -5-yl)thio-__ __ m~yl-3-cephem-4-carb-oxyl-te ~. ' , .
The tetrahydrofuran solution prepared above was cooled to -78C and a methanol solution of . ~ . .

:;., . ' :
.~' ' ..' ' ' .

~'7~

lithium methQxide, prepared from 24 rng of ltthiurn and 2 ml oE methanol wa~ added th~relo. The reaction mixture was stirred at -7BC for 30 minutes and then trea~ed with 0.25 ml of acetic acid. The soltuion was poured into water and extracted with ethyl acetate. The combined extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was then evaporated o-ff under reduced pressure. The residue was purified by chromatography on silica gel, giving 220 mg of benzhydryl 7~-(2'-chloro-l'-methoxyethylidenimino)--7~-methoxy-3-(1-methyl-1H
tetrazol-5~yl~thiomethyl-3-cephem-4-carboxylate, as a powder.
. ' .
IR spectrum ~Nujol-Trade Mark) ~maxCm ~ 1775, 1730, 1650 NMR spectrum (CDC13) ~ ppm = 3.38 (3H, singlet), 3.50 & 3.60 (2H, AB
quartet, J = ~8 Hz~, 3.70 (3H, singlet), 3.73 (3H, singlet), 4.0B & 4.22 (2H, AB
quartet, J = 7 Hz), ; 4-12 & 4.43 (2H, AB
quartet, J =12 Hz), 4.96 (lH 9 singlet), 6.86 (lH, singlet), 7-15 - 7.45 (10~l, multiplet).
.

' .
:

~Z~4~~

(C') Be~ l 7R~-chloroa(.et~m:i.dr-7rr-rlle_ho~ 3~
_ . _ (I-met}lyl-lH-te~ Q~ hlolrlethyl-3-ce~
fi-carboxyla-te 120 mg of benzhydrul 7~-(2'-chloro-1'-methoxy--: 5 ethylidenimino)-7~-methoxy-3~ methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylate were dissolved in 4 ml of chlo.roform containing 0005 ml of quinoline and 0.5 ml of trime-thylchlorosilane. The reaction mixture was stirred at room ternperature (1C) over night, poured into water and extracted with chloro-form. The combined extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was then evaporated off under reduced pressure, The , residue was chromatographed on silica gel, giving 80 mg of benzhydryl 7R-chloroacetamido-7~-methoxy-3-(l-methyl-lH tetra2:ol-5-yl)thiomethyl-3-cephem-4-carboxylate.

IR spectrum (Nujol-Trade Mark) ~ max cm~l ~ 1770, 17303 1650 NMR spectrum (CDC13) ~ ppm - 3.47 & 3.57 (2H, AB
quartet, J = 18 Hz), ` A.
3 D 52 (3H, singlet), 3.78 (3H, singlet), `- 4.07 (2H, singlet), 25 4.27 & 4.42 (2H, AB
,: , quartet, J ~ 12 H~), ' I
.
.`:. ~37~
. . I
; ., ~f ~ 1, ', ' :' 5'~

5-01 (1l1 s.inyl.et~, &.88 ~lH, singlet), 7.15 -7.50 (lO~T, multiplet).

EXAMPL~ 6 (A) 7~-(2'-chloro-l'-methoxye_~lidenimino)-7~-methoxy-3-methyl-3-ce~hem-4-carboxylic ac,id : 0.5 ml of trimethylchlorosilane and 0.17 ml of triethylamine were added to a solution of 325 mg of 1 7-dichloroacetamido-3-methyl-3-cephem-4-carboxylic ' acid in 12 ml of chloroform. The reaction mixture was stirred overnight at 10C, giving a chloroform solution of trimethylsilyl 7-dichloroacetamidG-3-methyl-3-cephem-4-carboxylate.

15 , . This solution was added dro,pwise at -50C to a , mixture of 220 mg of phosphorus pentachloride, 0-15 '~ ml of quinoli.ne and 15 m]. of chloroform. The reaction mixture was then stirred at -50C for 3 hours and diluted with 20 ml of dry tetrahydrofuran.
To thi.s solution, at -78C, was added a methanol , solution of lithium methoxide prepared from 150 mg ,~ of lithium and 15 ml of methanol. I'he mixture was stirred at -78C for 30 minutes and then treated with 1.5 ml of acetic acid. The solution was poured . ~ .
-3~-' r~ 7~5~1 :into water and extracted with chloroform. I'he~
cGmbined extracts were washed with water and dried o-ver anhydrous sodium sulphate; the solvent was -then evaporated oEf-under reduced pressure.
The residue was dissolved in a small quantity of chloroform, to which was added excess hexane, to precipitate the desired compound. This process was repeated three times, giving 240 mg of 7R~(2'-chloro~ methoxyethylidenimino)~7 methoxy-3-methyl-3 cephem-4-carboxylic acid.

NMR spectrum (CDCl3) ~ ppm - 2,05 (3H9 singlet), 3.03 & 3.43 (2H, AB
quartet, J ~ 18HZ), 3.40 (3H, singlet), ~5 3.75 (3H, singlet), 4.18 & 4.53 (2H, AB
quartet, J = 12 H2), 5.00 (lH, singlet).
,, ' .

(A') 7R-(2~-chloro-1~-methoxYethYlidenimino)-7~-; 20 methox~-3-methyl-3-cephem-4-car _xYlic acid .;. ......
325 mg of 7-dichloroacetamido-3-methyl-3-cephem-4-carboxylic acid were suspended in 5 ml of dry chloroform containing 0.17 ml of triethylamine - and 0.085 ml of acetyl chloride. The reaction mixture was stirred, whilst cooling with ice-water, for 1 hour to give the corresponding anhydride.

.
,' ~39-~::

:, , r ~7~5~

Th? resulting so~lltion was then added at -22C Lo ; a mixture prepared by the addition of 0-18 ml of quinoline to a solution of 271 mg of phosphorus pentachloride in chloroform. The re~ction mixture was then stirred at -22C for 3 hours. 'I'o the resulting solution was added, at -22C, a methanol solution of lithium methoxide prepared from 80 mg of lithium and 7 ml of methanol. The mixture was stirred at -22C for 10 minutes, treated wi-th 0.8 ml of acetic acid, poured in-to water and extracted with chloroform. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was then evaporated off, giving 300 mg of crude 7~-(2'-chloro-1'-methoxyethylidenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylic acid~ This compound was used without purification in the next step of the reaction. However, if desired, a pure sample may be obtained by the procedure of Example - 6(A~.

The process described above was repeated, except that -the acetyl chlori.de was replaced by monochloroacetyl chloride; the same results were achieved.

!
..
...
., ~o .
.

s~:~

(B) 7~ c~ rn ~vl-4-carb~y~

I ml of trimethychlorosilane was adcled to a solution of 300 mg of 7R-(2'-chloro~ methoxy-ethylidenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylic acid in 1 ml of dry chloroform. The reaction mixture was stirred overnight at room ; temperature. The solution was then poured into water and extracted with ethyl acetate. The organic extract was then extracted with a buffer solution of sodim dihydrogen phosphate and and dipotassium hydrogen phosphate(pH 7.5).
The aqueous extracts were acidified with 5N
hydrochloric acid to pH2 and again extracted with ethyl acetate. The combined extracts were washed with water and dried over anhydrous sodium sulphate, after which the solvent was evaporated of~ under reduced pressure, giving 240 mg of 7~-chloroacetamido-7~-methoxy-3~methyl-3-cephem-- Z0 4-carboxylic acid as a powder.

. ..
IR spectrum ~Jugol-Trade Mar~) ~ ~ max cm ~3250, 1765~ 1595 :' .
NMR spectrum (CDC13) ~ ppm ~ 2.20 (3H, singlet), 3.23 (2H, singlet), 3 55 (3H, singlet), 4.13 (2H, singlet), : ;
,:-:; .
i " ' ' .,: . .

t7'~S~l 5.06 (IH~ sin~let), - 7.70 (IH, singlet), 8.62 (lH, singlet)0 ; EXAMPLE 7 (A) 3-Ace_~y~ y1-7~-(2'-chlorO~~ Ey=
ethylidenimino)-7~-methoxy-3~cephem-4-carboxylic acid .~ .
~ 0.4 ml of trimethylchlorosilane and 0.17 ml - of triethylamine were added to a solution of 383 mg of 3-acetoxymethyl-7-dichloroacetamido-3-cephem-4-carboxylic acid in 12 ml oE chloroform.
The reaction mixture was stirred a-t room temperature for 1-5 hours to give a solution of trimethylsilyl 3-acetoxymethyl 7-dichloroacetamido-3-cephem-4-~ 15 carboxylate.
,' This solutlon was added, at -50C, to a mixture prepared from 220 mg of phosphorus pentachloride, 0.15 ml of quinoline and 15 ml of chloroform. The mixture was stirred at -50C
for 3 hours and dilùted with 20 ml of tetrahydro-furan. To the resulting solution was added, at -78C, a methanol solution o-f lithium methoxide prepared from 150 mg of lithium and 15 ml of methanol. The reaction mixture was stirred at ~;
`~

,:, ~'7~S't~
-78C for 30 minu-tes ancl the~n -treated with L.5 ml of acetic acid. 1`he solu-tion was then poured in-to ~ater and extracted wi-th chloroform. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was then evaporated off. The residue was dissolved in a small amount of chloroform, from which the desired product was precipitated by the addition of excess hexane.
This process was repeated three times, giving 320 m of 3-acetox~7methyl~7~-(2'-chloro-l'-methoxyeth~liden-imino)-7~-methoxy-3-cephem-4-carboxylic acid.
, i .

NMR spectrum ~CDC13) ~ ppm = 2.05 (3H, singlet), - 3.2 - 3.7 (2H), - 3.44 (3H, singlet), 3.78 (3H, singlet), ; 4.13 & 4.46 ~2H, AB
- quartet, J = 13 Hz), 4~8 ~ 5.2 (2H), 5.02 (lH, singlet)O

(B ? 3-Acetoxymethyl~7~-chloroacetamido-7~-metho 3-cephem-4-carboxylic acid - 1 ml of trimethylchlorosilane was added to asolution of 280 mg of 3-acetoxymethyl 7R-(2'~chloro-1'-methoxyethylidenimino)-7cy-methoxy-3-cephem~
carboxylic acid in 10 ml of dry chloroform~ The reaction mixture was stirred overnight at room temperature and then poured into water and extracted .,,' ~, -~3-.' ~, , ' . ' .

S~Il w.ith ethyl acetate. The organic extracts were further ex[.racted ~ith a buffer solution of sodiwn dihydrogen phosphate and dipotassium hydrogen phosphate ~pH 7~5). The aqueous extracts were acidified to pH2 and 5N hydrochloric acid and then extracted with ethyl acetate. The combined ethyl acetate extracts were washed with water and dried over anhydrous sodiurn sulphateO
The solven-t was then evaporated off under reduced pressure~ giving 260 mg of 3-acetoxymethyl-7~-chloroacetamido-7~-methoxy-3-cephem-4-carboxylic acid as a powder.

~MR spectrum (CDC13) ~ ppm = 2007 (3H, singlet), 3.38 & 3.58 (2H, AB
~ 15 quartet, J = 18 Hz), : 3.62 (3H9 singlet), ; 4.18 (2H, singlet), 5.00 & 5.19 ~2H, AB
quartet, J -15 Hz), 5.15 (lH, singlet), ; 7.70 (lH, singlet).
, . , -~4-, `
.
.
. ' EX.~IPI.E 8 (A) ?~-( 2'-chloro-1,' me hoxyethyl,idenimino)~7~-methoxy-3-(1-methyl-~,T~ tetrazol~5-yl)thiomethyl-3-cephem-4-carboxylic acid 0.32 ml of trimethyl.chlorosilane and 0.14 ml of triethylamine were added to a solution of 374 mg of 7-dichloroacetamido-3-(1-methyl-1H-tetrazol 5-y].) .~ thiomethyl-3-cephem-4-ca.rboxylic acid in 10 ml of chloroform and the r~action mixture was stirred at 5 - 7C for 1.5 hours, giving a solution of 7.-dichloroacetamido-3-(1-methyl~-lH-tetrazol-5-yl) thiomethyl~3-cephem-4-carboxylate.
' ::. This solution was added to a mixture of 176 mg -' ~ of phosphorus pentachloride, 0-12 ml of quinoline ~ 15 and 15 ml of chloroform at -50C. The reaction ,' mixture was then stirred at -50C for 3 hours and ' diluted with 20 ml of tetrahydrofuran, which had .~ been cooled to -78C. To this solution was added, at -78C, a methanol solution of lithium methoxlde prepared from 140 mg of lithium and 15 ml of `, methanol. I`he mixture was stirred at -78C for ; .
. 30 minutes and then treated with 1.2 ml of acetic . , acid, The solution was poured into water and extracted with chloroform. The resulting chloroform . . .
solu-tion was washed with water and dried over anhydrous sodium sulphate, after which the chloroform was : . :
. -45-.~

~ ~Q~S~

evap~Lated off, leaving a crude residue. The acqueous ]ayer from -the chloroform extraction was then again ex-tracted with ethyl acetate and the extracts were washed with water and dried over sodi~n sulphat~; the ethyl ace-tate solvent was removed by distillation under reduced pressure, giving a further residue. These residues were combined and dissolved in a ~small amount of chloroform, to which was added excess hexane to prec pitate the desired compound. This precipitation process was repeated three times, giving 200 mg of pure 7B-~2~-chloro~ methoxylid enimino)--7~-methoxy-3-(1-methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid.
.~ ~
NMR spectrum (CDC13) ~ ppm = 3.38 & 3.56 (2H, AB
quarte'c, J = 19 Hz), 3.46 (3H, singlet), 3n78 (3H, singlet) 5 3.82 (3H, singlet), 4.20 ~ 4.55 (2H, AB
quartet, J ~ 12 Hz), . .
4.47 (2H, singlet), -- 5.07 (lH, singlet), .''~ ' ;' ' ' . ..

-~6-lB) 7~-Chloroacetamido-7~ rn hoxy 3-(;-me~l~y~
lH--tetrazol-5-yl)thiomethyl 3-cephem-4-carboxylic acid 1 ml of trimethylchlorosilane was added to a solution of 300 mg of 7~-(2'-chloro-1'-methoxyethyl-idenimino)-7~r-methoxy-3-(1-methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid in 10 ml of chloroform~ The reaction mixture was stirred at room temperature (about 10C) overnight and then poured into water and extracted with ethyl acetate.
The resulting organic solution was extracted with disodium hydrogen phosphate solution (pH8). The aqueous extracts were acidified to pH2 with 5N
hydrochloric acid and again extracted with ethyl acetate. The ethyl acetate extracts were washed with water and dried over anhydrous sodium sulphate.
The solvent was evaporated off, giving 250 mg of 7R-chloroacetamido 7~-methoxy-3-(l-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid, as an amorphous powder~
`'; , .. , . ' IR spectrum (KBr) Y max cm 1 = 1760, 1680, 1605.
. ' NMR spectrum (CD3CN ~ D20) ~ ppm = 3.44 (3H, singlet)~
3.28 & 3.60 (2HJ AB
artet, J - ]8 Hz), 3.87 (3H, singlet), .

.

, -1~'7'~5~
4-11 (2H, singlct), 4.30 (2H, singlet), 5.01 (lH, slnglet).

7~-~2'-Chloro~ methoxyethylidenl no)-7~-methoxy-3-~1-methyl~ tetrazol-5-yl)thiomethyl-3~
cephem-4-carboxylic acid '.
435 mg of. 7-diachloroacetamido-3(1-methyl~
tetrazol-5~yl)thiomethyl-3-cephem-4-carboxylic acid were suspended in 5 ml of dry chloroform containing - 0.17 ml of triethylamine and 0.085 ml of acetyl chloride. The reaction mixture was stirred, whilst cooling with ice-water, for 1 hour, at tne end of which time the resulting solution was diluted with 14 ml of chloroform and 0.18 ml of quinoline and cooled to -22C. 271 mg of phosphorus pentachloride were then added and the mixture was stirred at -22C for 3 hours 7 cooled to -78C and diluted with 20 ml of tetrahydrofuran. To the resulting so].ution was added, at -78C, a methanol solution of lithium methoxide prepared from 80 mg of - lithium and 7 ml of methanol. The reaction mixture was stirred at the same temperature for 20 minutes and treated with 0.8 ml of acetic acid. The solution was then poured into water and extracted wit.:h chlorororm~ The chloroform extracts were washed with water and dried over anhydrous sodiurn sulphate, after which the chloroform was evaporated off, giving 370 mg of 7~-(2'-chloro-1' methoxyethy-lidenimino)-7~-methoxy-3-(1-methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid as a crude ~roduct.

- This crude product was used to prepare 7~-chloroacetamido 7~-methoxy 3~ methyl-1H-tetrazol-5-yl)thiomethyl 3~cephem-4-carboxylic aci.d by the procedure described in Example 8(B). ~ pure sample was also ob-tained by dissolving the crude : product in a small amount of chloroform and adding ' ' ; excess hexane to precipitate the compound.

.;` ' .
` 7~-(2'Chloro=l'-methoxyethy]idenimino)-7~-methoxy-3-(l-methyl-lH-tetrazol-5--yl)thiomethyl-3-. cephem-4-carbox ~

., , 435 mg of 7-dichloroacetamido-3-(1-methyl-lH-;~ 20 te-trazol-5-yl)thiomethyl-3 cephem-4-carboxylic acid were suspended in 5 ml of dry chloroform containing .~ 0.17 ml of triethylamine and 0.8 ml of monochloro-;~ acetyl chloride. The reaction mixture was stirred, whilst cooling with ice-water, for 1 hour~ The - ~

4~, procedure descrlbed in Example 9 was then repeated, except that the solution of llthium methoxide in methano] was added at -22C, instead of -78C, giving 350 mg of 7~-(2'-chloro-~'-methoxyethyl-i.denimino)-7~-methoxy-3-(1-methyl-1H-tetrazol-5-yl)-thiomethyl-3-cephem-4-carboxylic acid, as a crude product.

This was used, as described in Example 8(~), to produce 7~-chloroacetamido-7~-methoxy-3-(1- -methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.

. .
; EXAMPLE 11 .

(A~ ?R-(-2~-chloro-l~-methoxyethyl _enimino)-7~-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-` 15 cephem-4-carboxylic acid 6.6 g of 7-dichloroacetamido-3-(1-methyl-lH~
tetrazol~5-yl)thiomethyl-3-cephem-4-carboxylic acid : were suspended in 300 ml of anhydrous chloroform and the suspension was then converted to a homo-geneous solution by the addition of 2.5 ml of triethylamine~ 5.7 ml of trimethylchlorosilane were then added to the solution and the mixture was stirred for 30 minutes and then allowed to stand overnight at room temperature. After adding 4.5 ml of quinoline, the mixtu~e was cooled to , . ' , ~t~'~5 ~5()C and 3.8 ~ of phospllo:us pentachlo.ride.wer~
adcled. The solution was Ihen ~tirred for 3.5 hou:rs at -50C, whereupon it became clear. 1 ml of dry tetrahydrofuran were added to the so]ution andJ af-ter cooling to -70C~ the mixture was stirred at a temperature be-tween -60C and -70C
for 30 minutes with a methanol solution of lithiurn methoxide prepared from 1-5 g of lithium and 120 ml.
. of methanol. 6.5 ml of acetic acid in 15 ml of . tetrahydrofuran were then added. The solvent was distilled off under reduced pressure, giving c.rude ;; ; 7~-(2'-chloro~ methoxyethylidenimino)-7~-methoxy~
-~ - -3-~1-methyl-lH-tetrazol-5-yl)thiomethyl-3~cephern-4-carboxylic acid, which was used directly in the next step.
.~ , .
: (B) 7R-Chloroacetamido-7~-rnethoxy-3-(~-1nethyl-lH-tetrazol-5-yl)thiomethy]-3-cephem-4-carboxylic acid .', , .
~: The crude product obtained above was suspended in 200 ml of anhydrous chloroforrn and 30 ml of . 20 trimethylchlorosilane were -then added. The mixture ~; was stirred overnight at room te~perature, after -which the solvent was distilled off under reduced pressure. The residue was extracted with phosphate buffer (pH7.8), and the extracts were washed with ethyl acetate, adjust~d to pH2.1 by addition of dilute hydrochloric acid and then extracted with .. ethyl acetate. The extracts were washed with water ... .
i:, ~ -51-.

and dried~ af-ter which -the solvent. was c!istl].led off under reduced pressure. The residue was disolved in a small volume of ethyl ace-tate and the solution was added to hexane. The precipitates produced were collected by filtration, to give 4.6 g of 7~-chloroacetamido-7~-methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4~carhoxylic acid.
.~ ~

~ EXAMPLE ~2 ' ~ ... .
.
; 10 ~ 7~-(2'-chlor~'-methoxyethylidenimino)~7~-; methoxy-3-(1-methyl-LI~t.etrazol-5-yl)thiornethyl-3-cephem-4-carboxylic acid 3.8 g o:E 7-dichloroacetamido-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid were suspended in 400 ml of anhydrous chloroform and a homogeneous solution was then formed by the addition of 3.3 ml of trie~hylamineO
2.4 ml o:E dichlorodimethylsilane were added to the soluti.on and the mixture was stirred for 2 hours at room temperature. 6 ml of quinoline were added and the mixture was cooled to -50C, after which 5.0 g of phosphorus pentachloride were added and the mixture was stirred at -50C for 3.5 hours.
The solution became clear. Dry tetrahydrofuran was added to the solution in an amount of 100 ml :.,' , ~ . . . . . .

\`

and, atter cooling to -70C, the mixture was st.irred, a-t a temperature between -60C and -70C, for 30 minutes with a me-thanol solution of lithium ; methoxide prepared from 2.0 g of lithium and 150 ml of methanol. 8~7 ml of acetic acid in 15 ml of ; tetrahydrofuran were then added. The solvent was distil.led off under reduced pressure, yiving :. . crude 7~-(2'-chloro~ methoxyethylidenimino)-7~-; methoxy-3~ methyl-lH-tetrazol-5-yl)-thiomethyl-3 -cephem-4-carboxylic acid, which was used directly . in the next step.

.~ (B) 7~-Chloroaceta.mido-7~-methoxy-3-(1-methyl-1E~tetrazol-5-yl)thi.omethyl-3-cephem-4-carboxylic acid 'rhe crude product obtained above was suspended in 270 ml of anhydrous chloroform, and 40 ml of ~. dichlorodimethylsilane were added. The mix.ture ~ was stirred overnight at room temperature and thesolvent was then distilled off under reduced pressure. The residue was extracted with phosphate . 20 buffer (pH7.8) and the extracts were washed with . ..
ethyl acetate, adjusted to pH2.1 by addition of . dilute hydrochloric acid and extracted with ethyl i aceta-te. The extracts were washed with water and i dri~d and the solvent then evaporated off under :~ 25 reduced pressure. Theresidue was dissolved in a ~ small volume of ethyl acetate and the solution was .- added to hexane, which caused the precipitation of ... .

.
: -53-:
;', 6.6 g Gf 7R-chloroacetamido-7~methoxy-3-(l~
methyl-~ tetrazol-5-yl~thiome~thyl-3-cephem-4-carboxylic acid.

EX~ PIE ~
7'~-Chloroacetamido-7~-met xy-3-(1-methyl-tH~tetrazol-5-yl)thiomethyl~3-cephem-4-carbox~lic acid 1.32 g of 7-dichloroacetamido-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid were suspended in 18 ml of anhydrous chloroform.
0.85 ml of triethylamine was added, with ice-cooling, ~, .
to the suspension and the mixture was then-stirred for 5 minutes. To the mixture was then added 0.27 ` ml of phosphorus trichloride, after which stirring continued, with ice-cooling, for 30 minutes. After cooling the mixture further to -20C, 1.42 ml of dry quinoline and 1-26 g of phosphorus pentachloride were added and the mixture was stirred for 2 hours at -20C. 18 ml of dry tetrahydrofuran were added t-o the mixture, which was then cooled with dry ice/
acetone.

., A methanol solution of lithium methoxide prepared from 315 mg of lithium and 25 ml of methanol was added to the mixture, whilst maintaining the internal tempera-ture below -50C. The mixtuxe was .

:
' .
:, .
, ~J~541 then stirred for 1 hour, after which 3ml of ace-tic acid were added and stirring was continued for a further 1 minutes. The solvent was then distilled oEE under reduced pressure and the residue was dissolved in 30 ml of anh~rdrous chloroform; this solution was then evaporated to dryness under reduced pressure.

The residue obtained was suspended in 30ml of anhydrous chloroform containing 6 ml of tLime-thylchlorosilane and the mixture was stirred overnight at room temperature. The solvent was then distilled off under reduced pressure. 100 ml of ethyl acetate and 50 ml of water were added to the residue and the mixture was vigorously agitated. The separated aqueous layer was extracted with 100 ml of ethyl acetate. The combined extracts were re-extracted with 50 ml of a buffer solution (ph7.5, 10% w/v KH2P0~ : 10% w/v Na2HP0~ = 5l)-The pH of the aqueous layer was then adjusted to 2.0 by addition of 10% w/v HCl, whilst coolin~
with ice-water. The mixture was then saturated with sodium chloride and extracted three times ~with 100 ml of ethyl acetate. The cornbined extracts were dried over anhydrous sodium sulphate and the solvent was distilled off. 5 ml of ether were added -to -the residue and the mixture was washed with 10 ml of n-hexane and -the solvent was "

~ -55-"

%54~
.- ~

~ removed by decantat:ion. The residue was dri.ed .' under reduced pressure giving 450 mg of 7R-chloroacetamido-7~~methoxy-3~ me-thyl-1H
tetrazol-5-yl)thiomethyl-3-cephem-~-carboxylic . 5 acid was a pale yellow powder.

.~`

EXP*lP~E l4 (A) ~-Bromophenacyl 3-acetoxymethyl-7R-(2' bromo~ chloro-2'-methylthioethylJdenimino~-3-cephem-4-carboxylate .. ~

~ 10 A tetrahydrofuran solution of this compound `. was prepared from 330 mg of ~-bromophenacyl 3-acetoxymethyl-7R-(2'-bromo-2'-methylthioaceta-miclo)-3-cephem-4-carboxylate, 180 mg of phosphorus : .
pentachloride and 0.13 ml of quinoline, following ~: 15 the procedure described in Example 1.

. (B) ~-Bromophenacyl 3-acetoxymethyl-7~-methox~
7~ -methoxY-2~-me-t-hylthlothyl-idenimino~-3 cephem-4-carboxylate ,.,~
; A methanol solu-tion of lithium methoxide -.~ 20 prepared from 35 mg of lithium and 3 ml of methanol was addecl, at 70C, to the tetrahydro-., .~ furan solution of p-bromophenacyl 3-acetoxymethyl-7~(2'-bromo-1'-chloro-2'-methylthioethylidenimino)-:' :

.

:, %541 3-cephem-4-carboxylate prepared above~ The reactioll mixture was stirred at -70C for 30 minutes and then treated with 0.25 ml of acetic acid. The solution was poured into water and extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent wàs then evaporated off under reduced pressure. Th~ residue was purified by chromatography on silica gel, giving ~20 mg of ~-bromophenacyl 3-ace-toxymethyl-7~-methoxy-7~-~ methoxy-2~-methylthioethylidenimino)-3-cephem 4-carboxylate.

NMR spectrum (CDC13j ~ PP~I = 2.05 (3H, singlet), 2.18 (3H, singlet), 3.45 ~3H, singlet)-, 3.3 - 3.9 (4H, two types of AB quartet), 3.78 (3H, singlet), - 5.02 (2H and ~H), 5.45 ~ 5.55 (2H, AB
quartet), 7~55 - 7,90 (4H, _multiplet).
.
(C) p-Bromophenacyl 3-acetoxymethyl-7~methoxy-25 -meth~lthioacetamido-3-_ephem-4-carbox~late -1 ml of quinoline ancl 0.5 ml of trimethyl-~ .

4~

chlorosilane were ad~ed to a solution oE 100 mg of p-brGrnophenacyl 8-acetoxymet:hyl-^7~-methoxy-7~--( 1 ' -rne thox~r-2 ' -me thyl thioethylidenimino ) -3-cephelrl-4-carboxylate in 10 ml of dry chloroform. The reaction mixture was stirred overnight at room temperature. The solution was then poured into water and extracted with ethyl acetate~ The extracts were washed with water and dried over anhydrous sodium sulphate and the so]vent then evaporated off, giving 80 mg of ~-bromophenacyl 3-acetox~methyl-7~methoxy-7~-methylthioethyli-denimino-3-cephem-4-carboxylate.
.

IR spectrum (liquid) max cm = 1780, 1740, 1705 :
.
"

., ' .
~' .

-5~- , s~

, .
(A) p-Bromophenacyl 3-~ceto~ymethyl-7~-methoxy-7B-methoxy-2' ph~ i e~ylidenimino) 3-cephern-~-carboxylate .
Following the procedure described in Example 1, a te-trahydrofuran 301ution oE ~-bromophenacyl - 3-acetoxymethyl-7-(2'-bromo-1'-chloro-2'-phenylthioethylidenimino)-3-cephem-4-carboxylate was prepared from 350 mg of ~bromophenacyl 3-acetoxymethyl-7-(2' bromo-2'-phenylthioaceta-mido)-3-cephem-4-carboxylate~ 180 mg of phosphorus tetrachloride and 0.13 ml of quinoline. A methanol solution of lithium methoxide, prepared from 240 mg of lithium and 2 ml of methanol, was added, at -70C, to this tetrahydrofuran solution. The reaction mixture was then stirred at -70C for 30 minutes and treated with 0.25 ml of acetic acidO
The solution was poured into water and then extracted with ethyl acetate. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was evaporated off under ^~ reduced pressure, giving ~50 mg of ~-bromophenacyl 3-acetoxymethyl~7~-methox~-7B-(1'-methoxy-2'-- phenylthioe-thylidenimino)--3-cephem-4-carboxylate.
IR spectrum (liquid) Y max cm 1 = 1770, 1735, 1700.
;~ NMR spectrum (CDC13) ~ppm - 2.05 (3H, singlet), 3.30 & 3.55 (2H~ AB quartetJ J = 18 Hz), 3~36 (3J1~ sin~let), 3.70 (31i, singlet), 3.30 & 4.12 (2E~, AB quarte-t9 J = 13 E~z), 4.90 & 5.07 (2H, AB quartet, J - l4 l-lz) 9 5.00 (111, singlet), 5.35 & 5.52 (2H, AB quarte-t9 J ~ 14 Hz), 7,0 - 7.4 (5H), 7.4 - 7.9 (4H)~

, (B) ~-Bromophenacyl 3-acetoxymethyl--7~-methoxy-7~-phenylithioacetamido-3-cephem-4-carboxylate .
0.05 ml of quinoline and 0.3 ~m of trimethyl-chlorosilane were added to a solution of 80 mg of ~-bromophenacyl 3-acetoxymethyl-7~-methoxy-7~-~ methoxy-2~-phenylthioethylidenimino~-3-cephem -4-carboxylate in 5 ml of chloroform~ The reaction mixture was stirred at room temperature for 6 hoursJ
whilst nitrogen gas was introduced into the solution.
After pouring the solution into water, the resulting mixture was extracted with chloroform. The extracts were washed with water and dried over anhydrous sodium sulphate; the solvent was then evaporated off under reduced pressure. The residue was chromatographed on silica gel, giving 55 mg of ~~bromophenacyl 3-acetoxymethyl-7~-methoxy--7B-phenylthioacetamido-3-cephem 4-carboxylate.
NMR spectrum tCDC13) ~ ppm = 2.08 (3H, singlet), 3.27 & 3.40 (2EI, AB quartet, J = 18 Hz), .
. -~0-.

v~z~

3~37 (3I-I, singlet), 3.6~ (2H, singlet~, 5.05 (2H and lH).
5.46 (2H), 7.2 - 7.4 (5H, multiplet~ J
7.6 - 7.8 (4H, multiplet).

~ EXAMPIE 16 .' .
A) ~ _ 7~-(l'-methoxY-2~-~-thienylethylidenimino~-3 cephem-4-carboxylate Following the procedure described in Example 1.
a tetrahydrofuran solution of ~-bromophenacyl 3-acetoxymethyl-7~-(1', 2'-dichloro-2'-~-thienylethylidenimino)-3-cephem-4-carboxylate , was prepared from 315 mg of ~-bromophenacyl 3-acetoxymethyl-7R-(2'-chloro-2'-~-thienlacetamido)-3-cephem-4-carboxylate, 130 mg of phosphorus pentachloride and 0.13 ml of quinoline. A
methanol solution of lithium methoxide prepared from 24 mg of lithium and 2 ml of methanol was then added, at 70C, to this tetrahydrofuran - solution. The reaction mixture was stirred at ~70C for 30 minutes and then treated with 0.25 ml of acetic acid. The solution was poured into water and extracted with ethyl acetate. The ,, '' I

,'''' ~

~7~5~

exl-racts were washed ~ith water and dried o~er anhyclrous sodium sulphate J aEter which -the solvent was evaporated off under reduced pressure. The residue was puriEied by chromato-graphy on silica gel, yiving 160 mg of ~-bromo~
phenacyl 3-acetoxyrnethyl-7~-methoxy-7~-(1'-mekhoxy-; 2'-~-thienylethylidenimino)-3-cephem-4-carboxylat:e.
IR spectrum (Nujol-Trade Mark) ~ max cm 1 ~ 1780, 1740, 1705, 1650.
NMR spectrum (CDC13) ~ ppm = 2.10 (3H, singlet), ... .. .
3.43 (3H, singlet), 3.40 & 3.60 (2H, AB quartet, J ~ 18 Hz), 3.77 (3H, singlet), 4.03 & 4.30 (2H, AB quartet, J = 15 Hz), . .
5.0 - 5.3 (2H and lH), ' I 5.45 - 5.60 (2H J AB quarte-t, J = 16 Hz), 6.9 - 7.4 (3H, multiplet), 7.5 - 7.9 (4H, multiplet).
,~ ,, .

(B) ~-Bromophenacyl 3-acetoxymethyl-7~-methoxy-7~-(2'-~-thienylacetamido)-3-cephem-4-carboxylate , . ....
- 0-1 ml of quinoline and 0.5 ml of trimethyl-chlorosilane were added to a solution o-E 100 mg oE p-bromophenacyl 3-acetoxymethyl-7~-methoxy -` 7~-(1'-methoxy-2'-~-thienylethylidenimino)-3-cephem-4-carboxylate in 1 ml of chloroform. The reaction mixture was stirred at room -temperature overnight and then poured into water and extracted ~ -62-:

```~ 1~'7'~5~

with ethyl ace-ta-te. The extracts were washeh with water and dried oveL anhydrous sodium sulphateJ
after -which the solvent WaS evaporated off under reduced pressure, gi~ing 70 mg of ~~bromophenacyl 3-acetoxymethyl-7~-methoxy-7~-(2'-~-thienylaceta-mido)-3-cephem~4~carboxy]ate.
IR spectrum (Mujol-Trade Mark~ ~ max cm ~ ~ 1780, 1740, ~700, 3330-~R spectrum (CD3COCD3) ~ ppm = 2.07 (3H, singlet), 3.40 & 3.50 (2H, AB quartet, J = 18 Hz), 3.45 (3H, singlet), 3.87 (2H, singIet), 5.01 ~2H) 9 5.04 (lH, singlet), 5.45 (2H, singlet), 6.5 - 7.5 (3H), ( ~ 2 2 YP )-, , , .;
~A) Methyl 6~-(2'-chloro-1'-methoxyethylidenimino~-. ~ . . ~
~ 20 6~-metho y-penic anate .
Following the procedure described in Example 1.
a tetrahydrofuran solution of methyl 6~ ',2',2'-trichloroethylidenimino)-penicillanate was prepared from 300 mg of methyl 6~-dichloroacetamido~
penicillanate, 300 mg of phosphorus pentachloride . . , .

,., 1, ~qZS~L

and OG 2 ml o~ qu.i.noline. A rr,ethanol solution o:E
li-th:i.um methoxi.de prepared from 40 mg Or lithiwn . and 3 ml of methanol was adcled, at -70C, to ,., this tetrahydro-Luran solution. The reaction mixture was then s-tirred at: -70C for 30 minutes and treated with 0.5 ml of acetic acid. The . so].ution was poured into water and then extrac-ted ^ with ethyl acetate. The e~tracts were washed with water and dried over anhydrous sodium sulphate;
1 the solvent was then evaporated off under reduced :~ pressure. The residue was chromatographed on , silica gel, giving 170 mg of methyl 6~-(2'-chloro-1'-methoxyethylidenimino)-6~-methoxy-penicillanate~
IR spectrum (Nujol - Trade Mark) Y max cm~l ~ 1775, 1745, 1650.
NMR spectrum (CDC13) ~ ppm = 1045 (3H~ singlet), 1.52 (3H, singlet), . 3.50 (3H, singlet), : 3.78 (3H, singlet), 20 . 3.82 (3H, singlet), 4010 & 4.37 (2H, AB quar-tet, ~ = 13 Hz)g ~ .
;; 4.42 (lH, singlet), . 5~53 (lH, singlet)~
.'`'~ . I
. (B) Methyl 6~ chloroacetamido-6~-methoxy-:~ 25 penicillanate ~, 0.05 ml of quinoline and 0.5 ml of trimethyl-chlorosilane were added to a solution of 100 mg of '"

. -6~-., .' :

:;
,':

%~

- me-l-hyl 6~-(2' chloro~ methoxyethyliclenilrlino)-6~-metho~y--penicillclnate in L0 ml of dry chloroform The reaction mixture was stirred at l0C -Eor 6 hours. After adding water, the mixture was extracted wi-th ethyl acetate. The extracts were washed with ~ater and dried over anhydrous sodium su1phate; the solvent was then evaporated off under reduced pressure. The residue was chromatographed on silica gel, giving 45 mg of methyl 6~-chloroacetamido-6~-methoxy-penicillanateO
IR spec-trum (Nujol - Trade Mark) y max crn 1 = 3340, 1780, 1755, 1695~ -MMR spectrum ~CDC13) ~ ppm ~ 1-44 (3H, singlet), 1.53 (3H, singlet), i5 3.52 (3H, singlet), 3~77 (3H, singlet), ; 4.10 (2H, singlet), 4.47 (lH, singlet), 5.57 (lH, singlet), 7.67 (lH, broad singlet).
. .

.-.
(A) 7B-(2l-cyanomethylthlo-~-methoxyethyliden-imino)-7~-methoxy-3-(~-methyl-1H-tetrazol-5~yl) thiomethyl-3-cephem-4-carboxylic acid - 25 6 g o-f 7-(2'-cyanomethylthio-2'-chloroaceta-:

~, .

~'7Zs~i mi.clo)-3~(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-~-carboxylic acid, 2.18 ml of triethylamino and ~8 ml of trimethylchlorosilane were added to 180 ml of anhydrous chloroform. The mixture was stirred for 30 minutes and then allowed to stand overnight at room temperature. After adding 3.78 ml of quinoline, the mixture ~Jas cooled to -50C and . then stirred at this temperature for 3.5 hours with .. 3.42 g of phosphorus pentachloride. 60 ml of dry 1 tetrahydrofuran were added and the clear solution was then cooled to -78C and stirred at -78C for ~ 25 minutes with a methanol solution of lithi-~m methoxide prepared from l.2 g of lithium and 9G ml :- ~ of methanol. 5 ml of acetic acid in 15 ml of . 15 tetrahydrofuran were then added to the reaction ~ mixture~ The temperature of the mixture was ~: allowed to rise to room temperature, whilst .: stirring. The solvent was distilled off under .~ reduced pressure, giving crude 7~-(2'-cyanomethyl-thio-l'-methoxyethylidenimino)-7~-methoxy-3-(1 . methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-~-. carboxylic acid. This was used as a starting ~ material in the next step without purification.
"'' ' _ To illustrate the purification procedure, : 25 however, the above processes were repeated and .~ the product was then purified as follows: water ~ was added to the crude product and the mixture was :~ extracted with ethyl acetate; the extracts were ;; .

,. ' , .

~; ` `" lQ~Z5~1 ~ washed with water and dried and the solvent was ', evaporated off under reduced pressure; the residue ' was then dissolved in a small quantity of ethyl J acetate and precipitated with hexane, to give ; 5 4.2 g of the desired product in a pure state.
.. . .

(B) 7~-Cyanomethylthioacetamido-7~-methoxy-3-(l-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-_-carboxylic acid ;, - The crude product obtained above was suspended '; 1 in 168 ml of dry chloroform and 25.2 ml of trimethylchlorosilane were added to the suspension. ' The mixture was stirred overnight at room temperature and then the solvent evaporated off ; under reduced pressure. Water was added to the ,,~ 15 residue and the mixture was extracted with ethyl acetate. The extracts were re-extrac-ted with a phosphate buffer (pH7.8). The extracts were adjusted to pH 2.1 with dilute hydrochloric acid and then extracted with ethyl acetate. The '~ 20 extracts thus obtained were washed with water and dried and the solvent was evaporated off under , ,reduced pressure. The residue was dissolved in '' a small amount of ethyl acetate and then '~ ' precipitated with hexane, giving 3~6 g of 7~-cyanomethylthioacetamido-7~-methoxy~3-(1-methyl lH-tetrazol-5-yl)th:iomethyl-3-cephem-~-carboxylic ' acid.
.~

~67-l~;'Z5~-~
,,. ~

NMP spe(-trum (CD3COCD3) ~ ppm ~ 3~50 (3H, slnglet), 3.60 ~2H, singlet), about 3~5 - 3~7 (2H. qu~)rtet), 3.70 (211, singlet), 3.98 (3EI, singlet), 4.3 - 4.6 (2H, quarlet), 5-10 (lH, singlet)~

, 3 cetoxyme-thyl-7R-cyanomethylthioacetamido-7 methoxy-3-cephem-4-carboxylic-acid . . .
The procedure described in Example 18(a) was followed, except that the 7~(2'-cyanomethylthio-2'-chloroacetamido)-3-(1-methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem~4-carboxylic acid was replaced :. .
by 5.3 g of 3-aceto~methyl-7-(2'-cyanomethylthio-2'-chloroacetamido)-3-cephem-4-carboxylic acid, giving crude 3-acetoxymethyl-7~-(2'-cyanomethylthio-. ~
l'-methoxyethylidenimino) 7~-methoxy-3-cephem-4--~ carboxylic acid. This crude product was then 20 treated as described in Example 18(B), giving 3.2 g of 3-acetoxymethyl-7R-cyanomethylthioaceta-~; - mido-7~-methoxy-3-cephem-4-carboxylic acidO

NMR spectrum (CD3COCD3) ~ ppm = 2.07 (3H, singlet), about 3.5 (2H), 25 3.50 (3H, singlet), "' :
-6~ -~ 725 ,,; , 3 r 58 (~H, singlet), 3~73 (2~-1, single-t), 4,83 ~ 5.07 (2H, AB quarte-t, J ~ 13 Hz), 5.11 (lH, singlet).

EX~MPLE 20 (A) ~ nesulphonyl~ methoxyethyliden-imino)-7~-methoxy-3~ methyl-1H-tetrazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid . "
` ~ 562 mg of 7-(2'bromo-2'-methanesulphonylaceta-mido)-3-(l-methyl-lH-tetraæol-5-yl)thiomethyl-3-~;; cephem-4 carboxylic acid, 0.17 ml of triethylamine and 0.38 ml of trimethylchlorosilane were added to 12 ml of anhydrous chloroform. The mixture was stirred for 30 minutes and then allowed to stand at room temperature overnight. A*ter adding 0.6 ml of quinoline, the mixture was cooled to -~0C
and stirred at this temperature with 450 mg of phosphorus pentachloride for 3.5 hours. 20 ml ;~ of dry tetrahydrofuran were then added to the mixture. The mixture was cooled to -78C and ~;~ stirred at this temperature for 30 minutes with a methanol solution of lithium methoxide prepared from 110 mg of lithium and 10 ml oE methanol.
1 ml of acetic acid was then added and9 after `'"' ~ ~69-~ 54~

after adding w~ter, the mix-ture -was extract~d with chloroform, washed with water and dried;
the so]vent was then evaporated ofE, ~iviny 670 mg of crude 7~-(2'-methanesulphonyl~
; 5 methoxyethylidenimino)-7~-methoxy-3-~1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephern-4-carboxylic ; acid. This was used as the starting material i for the next stage without purification. If necessary, however, the crude product can be purified by recrystallisation from a mixture of - ethyl acetate and hexane.

Following the procedure described above, but ~ replacing the 7-(2'-bromo-2'-methanesulphonylace ;~ tamido)-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid with 7-(2'chloro-2'-methanesulphonylacetamido)-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4~carboxylic acid, the same product was obtained.
''' ' I

(B) 7~-Methanesulphonylacetamido-7~-methoxy-3-(l~methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4 carboxylic acid The crude product obtained abo~e was dissolved in 15 ml of anhydrous chloroform and the solutlon was stirred with 1 ml of trimethylchlorosilane overnight at room temperature. The solvent was distilled off under reduced pressuxe and, after , .

,' I
.

~ ~7Z5~

a~ding water, the mixture was extrclc-te~ wi-tll ethyl acetate The ethyl aceLa-le extrac~s were re-extrac-ted with a phosphate buffer (pH 7.8) and then the pH of the ex-tracts was adjusted to 2.1 by addition of dilute hydrochloric acid. I'he resulting aqueous solution was extracted with ethyl acetate. The extracts were washed with water and dried and then the solvent evaporated off under reduced pressure.
The residue was dissolved in a small amount of ethyl acetate 9 from which it was re-precipitated by addition of hexane. This precipitation procedure was repeated three times, giving 320 mg of 7B-methanesulphonylacetamido-7~-methoxy~3-(1~methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.
; MMR spectrum (CD3COCD3) ~ ppm = 3.07 (3H, singlet), 3D45 (3H, singlet), 3.62 (2H, quartet), 3.92 (3H, singlet), 4.18 (2H, singlet), 4.35 (2H, quartet), 5.06 (lH, singlet), 8 60 (lH, singlet).
~ , , .

, ' .

~7Z5 EX~Ml'~E 21 ~,~

7~-EthanesulE~onylacetamid -7~-me-thoxy_3-~
'', methy:L-~ tetrazo].-5-yl)thiomethyl~3-cephern-4-: arboxyl:ic acid The procedure described in Example 20~A) was repeated except that the 7-(2'bromo-methanesulphony~
lacetamido)-3-(1-methyl~lL tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid was replaced by 0.53 g of 7-(2~bromo-2~-ethanesulphonylacetamido)-3-( , 10 methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4 carboxylic acid; crude 7B-(2'-ethanesulphonyl~
, ~ methoxyethylidenimino)-7~-methoxy-3-(1-methyl-1H-. ~ tetrazol-5-yl.)thiomethyl-3-cephem-4-carboxylic acid ~ was obtained and was used directly as the starting ., . 15 material in the next step~ This crude product .,~ .
; was then treated as described in Example 20(~), giving 0.28 g of 7l-~-ethanesulphonylacetamido-7-methoxy-3-(1-methyl-lL-tetrazol-5 yl)thiomethyl-3-cephem-4-carboxylic acid.

.

., .
~, ~A) Methyl 7R-(2'=cyanomethylthio--l'- et~y~_~y~
lidenimino)-7~-methoxy-3-m_t~y~ arboxy-::
:- late To 10 ml of anhydrous chloroform were addc?d~

: . .
:

s~:~

:in succession, 2?0 m~ ol phosphorus pen-tachlortde and 0.17 ml of quinoline. 270 mg oE me-thyl 7-(2'-bromo-2'-cyanomethylthioacetamido)-3-~lethyl-3-cephem-4~carboxylate were added to the mixture.
The mixture was then stirred at room temperature for 40 minutes and the solvent-was evaporated off.
50 ml of dry tetrahydrofuran were added to the residue. The mixture was filtered and -the filtrate was cooled to -78C. The cooled filtrate was stirred at -78C for 20 minutes with a methanol solution of lithium methoxide prepared from 50 mg of lithium and 5 ml of methanol. 2 ml of water containing 0.5 ml of acetic acid were added to the reac-tion mixture to terminate the reaction~
After adding saturated aqueous sodium chloride solution, the mixture was extracted with ethyl acetate. The extract was washed with-water and dried and the solven-t evaporated off under reduced pressure to give 190 mg of methyl 7~-(2' cyanomethylthio-1'-methoxyethylidenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylate.
NMX spectrum (CDC13~ ~ ppm - 2.15 (3H, singlet), 2.95 & 3.20 (2H, AB quartet), 3.38 (3H, singlet), 3069 (3H, singlet) 7 3.74 (3H 9 singlet), 3~5 - 3.8 (4H), 4-93 (lH, singlet~.

7~

(B) ~lelhy:L 713~c~anomethylthioacetamldo _~Y-methoxy-;: ' .
350 rng of methyl 7~-(2'-cyanomethylthio~
mel:hoxyethyliclenlmino)-7~-mechoxy-3-methvl.-3-cephem -4-carboxylate, followed by .1 ml oE quinoline and 1^0 ml of trimethylchlorosilane J were added to l0ml oE anhydrous chloroform. The mixture was allowed to stand overnight at room temperature ancl then ;~ the solvent evaporated off under recluced pressure~
After adding water, the residue was extracted with , ethyl acetate. The extract was wa.shed with water ,~ and dried and the solvent evaporated off. The ; i , . .
residue was purified by silica gel chromatography, giving 280 mg of methyl 7~-cyanomethylthio-acetamido-7~-methoxy-3-methyl-3~cephem-4-carboxylate.
`;
NMR spectrum (CDC13) ~ ppm i=i 2.15 (3H, singlet)9 3.25 (2H), 3.48 (2H, singlet), 3.54 (5H, singlet), .. ! 20 3.82 (3H, singlet), 5.06 (lH, singlet), 7.87 (lH, singlet).

~' , ;.

, ' '' ,~"
. .

, ' ..

l~)'lZ5~
;~;
EX~MP~E 23 (A) ~ ~R~ b'~
lid~ mlno)~7~-me,thoxy-3-mel-.hy:L-3-cephem-4--carbox~rl~lte ; 3~10 mg of phosphorus pentach]oride iollowed by 0.23 ml of quinoline were added to 10 ml of anhydrous chloroformO 350 mg of methyl 7-(2'-bromo-2' methanesulphonylacetamido)-3-methyl-3 cephem-4-carboxyla-te were then added to the mixture). The ; mixture was stirred at room temperature for 50 minutes and then evaporated to dryness. To the residue were added 50 ml of dry tetrahydrofuxan and the mixture was filtered. The filtrate was cooled to -73C and ' then stirred at -78C for 20 minutes with a methanol solution of lithium methoxide prepared from 5G mg of lithium and lO ml of methanol. The reaction was terminated by adding 2 ml of water containing 0~4 ml of acetic acid. This mixture was then poured into water saturated with sodium ch]oride, ; and was extracted with ethyl acetate. The extracts were washed with water and dried and the solvent evaporated off under reduced pressure, giving 320 mg of methyl 7~-(2'-methanesulphonyl-1'-methoxyethylidenimino)-2~~methoxy-3 methyl-3-cepnem-4-carboxylate. The product can be purified hy silica gel chromatography.
NMR spectrum (CDC13)~ ppm = 2.04 (3H, singlet), 3.06 (3H, singlet), ~''`' , .

`

:. . .
.

S~Ll -~ 2.9 - 3.5 (211), 3.44 (3II, singlet), 3.78 (6H 9 singlet), 4.20 & 4065 (2H9 AB quartet, J ~ ~4 Hz), 4.97 (lH, singlet).
,' ' .
(B) Methyl 7~- _thanesu etamido-7~-methoxy- 3 -methyl-3-cephem-4-carboxylate ~ 300 mg of methyl 7~-( 2~-methanesulphonyl-l'-- methoxyethylidenimino)-7~-methoxy-3-methyl-3-cephem-4-carboxylate, 0.~ ml of quinoline and 1 ml of trimethylchlorosilane were added in turn to 10 ml of chloroform. The mixture was allowed to stand overnight at room temperature and then the solvent was evaporated off. After adding water, the residue was extracted with ethyl acetate. The extracts were washed with water and dried and the solvent was evaporated off under reduced pressure.
The residue was purified by silica gel chromato-graphy, giving 240 mg of methyl 7~-methanesulphony-lacetamido-7~-methoxy-3 methyl-3-cephem-4~carboxylate.
MMR spectrum (CDC13)~ ppm = 2.08 (3H, singlet), 3.08 (3H, singlet), 3.19 (2H), 3.49 (3H, singlet) 3.76 (3H, singlet), 4.07 (2H, singlet), 4-97 (lH, singlet), 8.33 (lH, singlet).
' -76 - .

,' :`
.,, ~ .

~`; iL0~2543L

. EX~I~- rll 2~

(A) ~ -2'--~.-th..l.eny~eth~
.lidenimino)-3-(~-methyl~H-te-trazol~5~y].)tlliomethJl~
~ hem-4-carboxylic acid 486 mg of 7-(2'-chloro-2'~-thienylacetalniclo)-3-(l-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid, 0.17 ml of triethylamine a~d 0.4 ml of -trimethylchlorosilane were added, in turn J
to 12 ml of dry chloroform. The mixtu.re was stirred at room temperature for 1 hour, after which 16 ml of dry chloroform and 0~3 ml of quinoline were added. The reaction mixture was then cooled ; to -60C and stirred at -50C for 305 hours with ~50 mg of phosphorus pentachloride. After addi.tion of 20 mm of dry tetrahydrofuran, the mixture was cooled to -78C, and a methanol solution of lithium methoxide, prepared from 1 mg of lithium and 1 ml of methanol, was added. The resulting miæture was stirred at -78C for 30 minutes and then the reaction was terminated by addition of 1- ml of acetic acid. Water was added and the mixture was then extracted with e-thyl acetate. The extrac-ts were washed with water, dried and evaporated to dryness under reduced pressure, giving 520 mg of crude 7~-methoxy~ rnethoxy-2~ thienyl-methylidenimino)-3-(1-methyl-1H-tetrazol-5-yl) t ; thiomethyl-3-cephem-4-carboxylic acid. The crude ." ~

.. i : .

.~

; -~O~Z5 . .

product was disso]ved :in a small volume of chloroform5 from which it was re-precipitated wi-th hexane to give a pure product.

(B) 7~ Methoxy-3-(1~methyl-1H tetrazol-5-yl) thiomethyl-7~ thienylacetamido)-3-ce~hem 4-carboxylic acid .
0.5 g of 7~-methoxy 7~-(1'-methoxy-2'-~-thienylethylidenimino)-3-(1-methyl-1H-tetraæol-5-yl)thiomethyl-3-cephem-4-carboxylic acid, 0.12 m]
of quinoline and 1.0 ml of trimethylchlorosilane were added, in turn, to 1 ml of chloroform. The mixture was stirred overnight at room temperatllre and then poured into water and extracted with chloroform. The extracts were re-extracked with - 15 a phosphate buffer (pH 7.5) and the aqueous layer ~;! was then extracted with ethyl acetate. The extracts were washed with water and dried and the solvent evaporated off under reduced pressure, giving 0.4 g of 7~~methoxy~3-ll-methyl-1H-tetrazol ~5-yl)thiomethyl-7~ -thienylacetamido)-3 cephem-4-carboxylic acid.
~- NMR spectrum (CD3COCD3)~ ppm = 3O42 (3H, singlet) 9 3.53 & 3.76 (2H, AB doublet7 J ~ 18 Hz), - 3.92 (2H, single-t), 3.96 (3H, singlet), 4.28 & 4.50 (2H, AB doublet, J = ~4 Hz), 5.0~ (lH, singlet), 6~8 - 7-1 (2H, multiplet), ~7~-~ Jz~

~27 (lH, broad single-t).
.

(A) Benzhydryl 3-acetox~le-thyl-7~-(2',2'-dichloro-L'-methox~e~ enimino)-7~-methoxy-3-ce~hem-4-~ y___e ; 0.13 ml of quinoline, followed by 291 mg of benzhydryl 3-acetoxymethyl-7-trichloroacetamido-3-cephem-4-carboxylate, were added, with ice cooling, to a solution of 180 mg of phosphorus pentachloride in 8 ml of anhydrous chloroform. The mixture was stirred, whilst ice cooling, for lhour, and the solvent was then distilled off under reduced pressure. To the residue were added 20 ml of anhydrous tetrahydrofuran and crystalline substances were filtered off~ The filtrate was ; stirred for 20 minutes at -78C with a methanol solution of lithium methoxide, prepared from 24 mg of lithium and 2 ml of methanol, and then 0.25 ml of acetic acid was added. The reaction mixture ~; 20 was poured into water, extracted with ethyl acetate, washed with water and dried; the solvent was then evaporated off, The residue was purified by silica gel chromatography, to give 30 mg of benzhydryl 3-acetoxymethyl-7~-(2',2 t ~ dichlorc-l~-methoxyethylidenimino)-7~-methoxy-3-cephem-4 ,' carboxylate.
IR spectrum (liquid film) J max cm 1 - 1770, 1730, 1650, N~ spectrum (CDCl3) ~ ppm = 1.38 (3H, singlet), 3.35 (3H, singlet), 3.2 - 3~5 (21I) 3.~0 (3H, singlet), 4D6 - 4~9 (2H ~- lH), 6.7 - 6.9 (lH ~ 1H), 7Dl - 7.5 (lOH)o Mass spectrum (m~c) ~ 596, 594, 5920 (B3 Benzhydryl 3-acetoxymethyl-7~-dichloroaceta-mido-7~-methoxy-3-cephem-4- arboxylate 0.1 ml- of quinoline and 0.5 ml of trimethyl-chlorosilane were added to a solution of 100 mg of benzhydryl 3-acetoxymethyl-7~-(2',2'-dichloro-l'-methoxyethylidenimino)-7~-methoxy-3-cephem-4-carboxylate in 5 ml of anhydrous chloroform~ The mixture was stirred at room temperature for 5 hours and then water was addedO The mixture was extracted with chloroform, washed with water and dried and the solvent evapora-ted to dryness under reduced pressure to give 60 mg of the desired product.
- ' .
, ~
',; , . .

... .

Claims (10)

THE EMBODIMENTS OF THE INVENTION IN WHICH
AN EXCLUSIVE PROPERTY OR PRIVILEGE IS CLAIMED ARE
DEFINED AS FOLLOWS:

1. A process for preparing a compound of formula (I):
(I) wherein R1 represents a hydrogen atom, a haolgen atom, an alkyl group having from 1 to 4 carbon atoms or an aryl group; R2 represents a hydrogen atom, a halogen atom, an alkyl group having from 1 to 4 carbon atoms, an aryl group, an alkylthio group having from 1 to 4 carbon atoms, an alkynylthio group having from 2 to 4 carbon atoms, an arylthio group, an azidoalkylthio group having from 1 to 4 carbon atoms, a cyanoalkylthio group having from 1 to 4 carbon atoms in the alkyl moiety, an alkyl-sulphonyl group having from 1 to 4 carbon atoms, a 5- or 6- membered heteorocyclic group, a 5- or 6-membered heterocyclicthio group, or a 5- or 6-membered heterocyclic-oxy group; R3 represents an alkyl group having from 1 to 4 carbon atoms;
and Z represents a fragment of formula:

or wherein R4 represents a carboxyl group, an alkoxycar-bonyl group having from 1 to 4 carbon atoms in the alkyl moiety, a haloalkoxycarbonyl group having from 1 to 4 carbon atoms in the alkyl moiety, a benzyloxycarbonyl group which is unsubstituted or has one or more halogen, methoxy or nitro substituents, a diphenylmethoxycarbonyl group, a trialkylsilyloxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety, a dialkylhalo-silyloxycarbonyl group having from 1 to 4 carbon atoms in each alkyl moiety, a phenacyloxycarbonyl group which is unsubstituted or has one or more halogen or nitro substituents, an acyloxycarbonyl group, a haloacyloxycarbonyl group, a dihalophos-phinoxycarbonyl group, a dialkylphosphinoxy-carbonyl group or an aminocarbonyl group; and A represents a hydrogen atom, an azido group or a group of formula - B-E wherein B represents an oxygen or a sulphur atom and E represents an acyl group, an alkyl group having from 1 to 4 carbon atoms or a substituted or unsubstituted carbomoyl, thiocarbamoyl or heterocyclic group which process comprises reacting a dialkoxyimino compound of formula (V):

(V) (in which R1, R2, R3 and Z are as defined above) with a halo-silane or an acid and treating the resulting product with water.

2. A process as claimed in claim 1, wherein said halosilane is a dialkyldihalosilane or a trialkylhalosilane.

3. A process as claimed in claim 2, wherein said dialkyldihalosilane is dimethyldichlorosilane, diethyldichlorosilane, dimethyldibromosilane or diethyldibromosilane.

4. A process as claimed in claim 2 wherein said trialkylhalosilane is trimethylchlorosilane, triethylchlorosilane, trimethylbromosilane or triethylbromosilane.

5. A process as claimed in claim 1 which comprises treating 3-acetoxymethyl-7.beta.-(2'-chloro-1'-methoxyethylideneimino)-7.alpha.-methoxy-3-cephem-4-carboxylic acid with trimethylchlorosilane in dry chloroform and then with water to produce 3-acetoxymethyl-7.beta.-chloroacetamido-7a-methoxy-3-cephem-4-carboxylic acid.

6. A process as claimed in claim 1 which comprises treating 7.beta.-(2'-chloro-1'-methoxyethylidenimino)-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid in suspension in anhydrous chloroform with trimethylchlorosilane and after removal of the solvent with water to produce 7.beta.-chloro-acetamido-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.

7. A process as claimed in claim 1 which comprises treating 7.beta.-(2'-cyanomethylthio-1'-methoxyethylidenimino)-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-car-boxylic acid in suspension in anhydrous chloroform with tri-methylchlorosilane and,after removal of the solvent,with water to produce 7.beta.-cyanomethylthioacetamido-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.

8. A process as claimed in claim 1 which comprises treating 3-acetoxymethyl-7.beta.-(2'-cyanomethylthio-1'-methoxyethyl-idenimino)-7.alpha.-methoxy-3-cephem-4-carboxylic acid in suspension in anhydrous chloroform with trimethylchlorosilane and after removal of the solvent,with water to produce 3-acetoxymethyl-7.beta.-cyanomethylthioacetamido-7.alpha.-methoxy-3-cephem-4-carboxylic acid.

9. A process as claimed in claim 1 which comprises treating 7.beta.-(2'-methanesulphonyl-1'-methoxyethylidenimino)-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-car-boxylic acid in suspension in anhydrous chloroform with tri-methylchlorosilane and,after removal of the solvent,with water to produce 7.beta.-methanesulphonylacetamido-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.

10. A process as claimed in claim 1 which comprises treating 7.beta.-(2'-ethanesulphonyl-1'-methoxy-ethylidenimino)-7.alpha.-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid in suspension in anhydrous chloroform with tri-methylchlorosilane and,after removal of the solvent, with water to produce 7.beta.-ethanesulphonylacetamido-7-methoxy-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.