Note: Descriptions are shown in the official language in which they were submitted.
)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 ~
--1--
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:
CH3
1 CH2A R4
.: 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--
~ 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.
.
--6--
~-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
-7-
(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):
~` ' ' :
'
.
: ~
--8--
~ 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).
--10--
. .
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)
: .
-12-
' '
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~
'~
.
:. .
'.
-14-
',
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
-15-
~'
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
',', :
. .
:,
.,
-16-
''.,'
'"'.
'
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
R2/ 1
.. ..
-17-
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
~.'
:`
., .
-18-
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
.,
,' '
-19-
.,
,' ,. .
(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.
. .
''
..
-20-
:,
:
^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
''
-21-
,
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
;'
~ .
~';
-23-
.
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'
,
,''
.' .
' '
'~:
:
, . . .
." ' .
.
:`
-24-
;"
.,' ~
~ ~ 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:
-25-
:~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
''
-26-
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
, .
-27-
~ ~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).
-28-
.
~7~5~
EXAMPLE 2
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.
EXAMPLE 3
(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
-29-
.
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.
- 30
:`
-30-
`~ 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
.
:
-31-
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).
EXAMPLE 4
(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
'
-32-
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),
-33-
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~),
:
-34
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),
EXAMPLE 5
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
. ~ . .
-35-
:;., . ' :
.~' ' ..' ' ' .
~'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).
.
-36-
'
.
:
~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
~;
`~
-42-
,:,
~'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),
.
.
- -47-
,
-1~'7'~5~
4-11 (2H, singlct),
4.30 (2H, singlet),
5.01 (lH, slnglet).
EXAMPLE 9
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.
EXAMPLE 1
.;` ' .
` 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
-49-
- ~
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-
:
;',
54
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
.
-5
:
' .
:, .
,
~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)-
:' :
-56-
.
:,
%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-
~ .
-57-
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~
FXAMPLE 15
, .
(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),
-59
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
-61-
,'''' ~
~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
. . ,
.
-63-
,., 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-
:
-65-
~, .
~'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
;; .
-66-
,. ' ,
.
~; ` `" 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
,
-70-
.
,' 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.
.
EXAMPLE 22
., .
~, ~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~
- 7 2 -
: . .
:
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).
~'
, ;.
74-
, ' ''
,~"
. .
, ' ..
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),
~''`' , .
-75-
`
:. . .
.
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).
.
EXAMPIE 25
(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
-79-
,' 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.
- ' .
, ~
',; ,
. .
... .
