Note: Descriptions are shown in the official language in which they were submitted.
~S3~
The present invention relates to a novel process
for the preparation of certain 7,B-acylamino-7c~-me$hoxycephalosporin
. derivatives.
In one known process for preparing 7,B-acylamino-
7~-methoxycephalo~porin derivatives, 7,B-(~-aminoadipo~l~amino-
. 7~-methoxycephalosporin compounds are produced by ferrnentation
and then acylated to form the corresponding diac~ ated compounds.
The aminoadipoyl group is then split off (see Japanese Patent
Provisional Publication No. 931/~2). The same Japanese I'atent
Publieation also describes a process in which a 7-amino-7-methoxy-
.i eephalosporin compound obtained by chemieal synthesis is aeylated
to give the corresponding 7~-acylamino-7O~ ethoxycephalosporir
~; : eompo~md. In these processes, the acylation at the 7~B-poi,ition- ~ ~ is effeeted by the use of an active derivative of the acid whose
. residue it Is desired to incorporate at the 7,B-position, such as
the acid chloride. II, however, the aeyl group to be introduced
,
derived from a substituted thioalkanoic aeid ~such as 1, 3, 4-thiadia-:
i, ~ zolylthioaeetic aeid or I,2,4-tria~olylthioacetic),this proeess .
is not praetical, since it :is often difficult or, in some cas.es,
impossible to prepare an active derivative OI sueh an acid; in
other cases, this active derivative may be unstable or may not
, , :
. ' ' .
~5~53~
be obtainable in a good yield. In any case, the yield of the acylation
reaction is extremely poor.
,
Another process is described in Japanese Patent
Provisional Publication No. ~;2791/73 In this process, a cephalosporin
compound, unsubstituted at the 7~- position, is converted to an
acylimine intermediate, which is then methoxylated to give the
desired 7~-methoxycephalosporin derivative, This process however,
also has disadvantages in that it cannot be carried out economically
and, in any case, still leaves unsolved the problem of introducing
the desired 7,B-acyl substituent in ~e starting material.
We hàve now discovered a process for the preparation
of 7cY-methoxycephalosporir. derivatives having a substituled thio-
~i alkanoylamino group at the 7~- position, which enables these
derivatives to be obtained under mild conditions, in very high
-~ ~ 15 yields compared with known processes and without any of the
disadvantages desFribed above. The process of the invention may
be used to prepare a wide variety of cephalosporin derivatives.
, . . .
Thus, in accordance with the present invention,
there is provided a process for preparing a 7~-methoxycephalosporin
, , ' . ':
-- 2
.
. ~ . , .. . .... , ... . . .... .. , ~ ..
3~
derivative having a 713 substituent of formula
R -- S--A-- CO.NH--
.
(in which: Rl represents a hydrogen atom, a substituted or
unsubstituted acyclic hydrocarbon group, a substituted or
unsubstituted monocyclic alicyclic hydrocarbon group having a
5- or 6-membered ring~ a monocyclic or bicyclic aromatic group
- (whose benzene ring may be substituted or unsubstituted), a
substituted or unsubstituted alkanoyl group, a substituted or
unsubstituted benzoyl group, or a substituted or unsubstituted
monocyclic or bicyclic heterocycli.c group having at least one
ring nitrogen, oxygen or sulphur atom; and A is a straight or
branched- chain alkylene group), wherein a 7~-methoxycephalosporin
5 ~ ' derivative having a 7~ substituent of formula
:. ~
,~ ~ X--A--CO,NH--
.
(in which: A is as defined above; and X is a halogen atom)
, IS reacted with a thiol compound of formula:
Rl _ SH
, . .
'. ,
'
.
,' ,~ ', ` ' :'
or with a metal salt thereof.
The 7~x-methoxycephalosporin derivative
}arepared is preferably a compound of formula (II):
OCH3
B -S--A- CO.NH ~S~ (11)
o N~S~ R
~, . .
R
(in which: R is as defined above; R 1S a group which does
not participate in the reaction, suc:h as those groups exemplified
below; and R3 is a carboxyl group or an esterified carboxyl group),
Examples of acyclic hydrocarbon groups
which may be represented by Rl are straight or branched- chain
hydrocarbon groups having from l to 6 carbon atoms, such as
methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl, pentyl,
hexyl, vinyl, aLlyl, propenyl, butenyl, pentenyl, hexenyl, ethynyl
or propargyl. These hydrocarbon groups may be unsubstituted
or they may be substituted with such substituents as: hydroxy
groups, azido group~; cyano groups; nitro groups; acylamino
groups, e.g. acetylamino; alkoxycarbonylamino groups, such
as as t -butoxycarbonylamino; alkoxy groups9 e.g. methoxy or
- 4 -
; ' ~ ' ' . .
.
~SiCI 53~
ethoxy; the phenyl group; the cyclohexadienyl group; the
cyclohexyl group; or alkoxycarbonyl groups, e,g. methoxy-
carbonyl or ethoxycarbonyl.
Where the group R represents a mono-
- cyclic or bicyclic aromatic group, it may be, for example, a
substituted or unsubstituted phenyl and naphthyl group,
Examples of suitable substituents include, for example: halogen
.
atoms, such as chlorine or bromine; alkyl groups, such as
methyl or ethyi groups, alkoxy groups, such as methoxy or
ethoxy; the cyano group; the nitro group; acylamino groups,
such as acetylamino; or alkoxycarbonylamino groups, such
as t-butoxycarbonylamino. If the group :Rl is a benzoyl group,
it may be unsubstituted or substituted and, if substitut`ed, the
~ substituents may be any of those exempllfied above with
lS respect to the phenyl and naphthyl groups.
Rl may also be a monocyclic alicyclic
group having a 5- or 6-membered ssturated or unsaturated
ring; examples of such groups include: cyclopentyl, cyclo-
I ~
penten~rl, cyclohexyl, cyclohexenyl and cyclohexadienyl.
2~ ~ Examples of alkanoyl groups, preferably having from 2 to 18
caFbon stomsj which may be represented by
,
~a~5DS3~
R include acetyl, propionyl, butyroyl and stearoyl groups,
which may be substituted or unsubstituted. If substituted,
suitable substituents include: cyano groups; nitro groups;
acylamino groups, e.g. acetylamino; alkoxycarbonylamino
groups, such as t-butoxycarbonylamino; and alkoxycarbonyl
gralps, e. g. ethoxycarbonyl or methoxycarbonyl.
Alternatively, Rl may be a monocycli c
or bicyclic heterocyclic group having at least one ring nitrogen,
oxygen or sulphur atom; the heterocyclic group may be
1~ aromatic or non-aromatic in character and examples include:
2-imidazolyl; 1, 2, 4-triazol-3-;yl; 1, 3, 4 -thiadiazol- 2-yl; 2-
pyridyl; 2-pyrimidyl; purin-6--yl; 2-benzothiaz~lyl; 2-
benzoxazolyl; s-triazolo[ 4,3-a] pyridin-3-yl; and 2-thiazolyl.
.
The group A is a straight or branched-chain alkylene
group, preferably a lower alkylene group having from 1 to 6 carbon
atoms, Examples include: methylene, trimethylene, propylene,
tetramethylene, 2-methyltetramethylene, pentamethylene and
hexamethylene groups.
The group R does not participate in
the reaction of the process of the present invention and its
nature is, there~ore, not critical to the present invention,
....
'
:, ,
,", , ~ , ~ , .. ...
, " . . .. . ... ..
5~;i3~
However, preferred groups R include: hydrogen atoms; lower
alkyl groups, e.g. methyl; acyloxymethyl groups, e.g. acetoxymethyl;
carbamoyloxymethyl groups; and thiomethyl groups which are
S-substituted by aromatic monocyclic heterocyclic groups,
S containing at least one nitrogen, oxygen or sulphur ring atom,
e . g . (l -methyl - lH - tetra zol - 5 - yl) thiom ethyl or (l, 3, 4 - thi adiazol - 2 -
yl)thiomethyl .
In formula (II), the group R is a carboxyl
or esterified carboxyl group. When R is an esterified carboxyl
group, the nature of the ester is not particularly critical, provided
that it does not destroy the cephem ring during or after the
condensation reaction and provided that it can readily be split
off from the cephem ring. Suitable esters include, for example:
alkylsilyl esters, such as trimethylsilyl esters; lower alkyl esters,
such as methyl, ethyl or t-butyl esters; benzyl esters; p-methoxy-
benzyl esters, ben~hydryl esters; phenacyl esters; ~-bromophen-
acyl esters; and 2, 2, 2-trichloroethyl esters. When R is an
esterified carboxy1 group, the process of the invention preferably
comprises the additional step of deesterifying the reaction
.
product,
- Where, as is preferred, the 7~ mFthoxycephalosporin
to be produced is a compoundof formula (II), defined above, the
,
- 7 -
:.
~53~
preferred 7~-methoxyceph~losporin starting material is a
compound of formula (III):
O~I3
~ S~ (III)
X - ~- CC~ 2
s~ N ~ R
.. O
`- !
;
~, .
(in which X, R and R are as deiined above)~. In this compoundJ
` X is a halogen atom, preferably chlorine or bromine.
'''
- The thiol compound of formula (I) may be
:`
employed as such or may be~ and preferably iB, employed in the
form of a metal salt, e. g. an all~ali metal or alkaline earth
metal salt, such as a sodium, potassium, lithium, calcium or
barium salt. Of these, the sodium, potassium and lithium salts
are preferred.
'' ' '.
The process c~f the present invention can
easily be carried out simply by contacting the 7a-metho~ycephalosporin
.,~ .
.
. .
i3~
derivative used as starting material, e. g. the compound of
formula (III) with the thiol compound of formula (I), or with a
metal salt thereoi. In order that the reaction should proceed
smoothly,~we prefer that it should be carried out in the presence
OI a solvent, particularly if a metal salt of the thiol (I) is employed.
Any solvent which does not participate in the reaction can be
used in the process of the present invention, including water and
~ . .
many organic solvents. Examples of suitable organic solvents
include dialkyl ketones, e. g. acetone or methyl ethyl ketone;
10 halogenated alkanes, e.g. chloroform9 methylene chloride or
dichloroethane; lower alkanols, e.g. methanol or ethanol; and
dimethylformamide. Mixtures of two or more solvents can also
be employed. If the 7Q-methoxycephalosporin derivative of
., .
formula (III) has a carboxyl group in the 4-position, we preîer
.
15 to employ a mixture of water and an organic solvent as the reaction
medlum.
The thiol (I) or metal salt thereof will
normally be used in an amount equimolar to the 7~-methoxycephalosporin
derivative or in a slight excess; thu~, for example, the molar
20 ratio of thioi (I) to ~a-methoxycephalosporin starting material
1: '
~: .
may be from 1: 1 to 1.1: 1.
., .
~ 9
.; ' .
-3
" .
~' .
.
- . ',' ',: :' ' ' ' '
', ,, ~.
:' ' ' ' , , ' ' i ' i
1~5~i3~
Although the reaction will proceed at
an approximately neutral pH, it proceeds more smoothly under
weak:Ly alkaline conditions. ~ccordingly, if the thiol (I) itself
iæ employed, we prefer to add an alkali to the reaetion mixture
in order that the reaction should be carried out under weakly
alkaline conditions. Suitable alkalis which may be used include:
sodium hydroxide, potassium hydroxide, sodium carbonate,
potassium carbonate, sodium bicarbonate and potassium bi-
earbonate.
The reaetion temperature is not
!~
eritical to the process of the invention and the reaetion is,
.: .
aeeordingly, normally effeetedat approximately ambient temperature,
` although the reaetion will proceed at temperatures higher
~f
and lower than ambient. The time required for the reaction
will vary dependingupon the reaetion temperature, the reagents
and other conditions, but the reaction will normally be eomplete
, . . .within ~rom 2 to 30 hours; if X in the 7a-rnethoxyeephalosporin
starting material is a ehlorine atom, the reactlon will normally
' take a relatively long time~ i. e. from 10 to 30 hours.
. , .
When the r eaetion is eomplete, the
- - ,.
.
-10 -
.
~L~50t~i3~
des;red compound, e.gO the compound of formula (lI),may
be recovered from the reaction mixture by conventional
methods. For example, if a carboxyl group is present in
the 4- position of the compound, the reaction mixture is first
acidified and, if a solvent other than water is employed for the
reaction, the solvent is removed by distillation, after which the
reaction product is extracted with a suitable solvent (e.g. ethyl
acetate) and the extract is washed with water and dried. The
solvent is then removed by distillation, giving the desired product.
.
If necessary, this may be further purified by conventional means,
.
e.g. by chromatography.
' If the compound pro~ucedby the pr ocess
of the invention has an csterified carboxyl group at the 4- positionJ
the reaction medium is, as stated above, preferably a mi~ture
1 S; of water and a miscible organic solvent. This organic solvent
ie flrst removed from the reaction mixture by distillation, after
which the residue is extracted with a water-immiscible solvent.
. . ~
The extract is then washed with water and the solvent removed
i ~y distillation to give the ester~fied compound. This compound~, 20 may be further purified by conventional methods, e.g.
chromatography, but, in general, we prefer that it should be
.
~[35C~3~
swbjected directly to deesterification to give the desired
compound. This deesterification can be carried out by
any conventional method, depending upon the nature of the
ester, Suitable deesterification processes include: alkaline
hydrolysis, acid hydrolysis and reduction with hydrogen.
AMer completion of the deesterification, the desired product
is reeovered from the reaction mixture in the manner described
.
above.
Many of the 7~-methoxycephalosporin
:. . .
derivatives of formula (II) have valuable antibacterial activlty
against various pathogenie bacteria. Representative exarnples
, of these compounds and their antibacterial activities (expressed
as m~nimiu~ ;nhibitory concentrations) are shown in the
following Table. Also shown, for purposes of comparisonj
~ are the minimum inhibitory con entrations against the same
baeteria of two known compounds, 3 - earbamoyloxym ethyl -
7~-methoxy- 7,B -phenylac etamido- 3 - c ephem -4 - carboxylic ac;d
,~ ~
'; and Cephalothin.
As will be seen from the Table, the most
, 20 preferred eompounds of the invention are those in which:R represents
a eyanomethyl, l-eyanoethyl, 2-hydroxyethyl, propargyl, azidomethyl or
3-isoxazolyl group; R represents a (l-methyl-iH-tetrazol-5-yl)thio-
methyl, earbamoyloxymethyl or acetoxymethyl group;
and A represents a methylene group.
' ' .
. .
1,
i3~L
~1 ~ ~
, .. _ ~_
h ¦ ~ ~ a~
~ . ~ ~
., ~ ~1 o o o c o
, ~ . . _ . , ~ . . ~.
Eoil ,~ I~ 1 Ic~
~ U~_ _ . .. . . _ . ~
..~ V ~ ~^ CO - .
. ~ ~ o~ :~o ~ CO,
: ~ ~ ~ V,~ " ~
,~ ~ ~, ~1 ~ ~ i ~ O
~ w wl ~o _ .-
_ C~ ~
~ m~ ~ m~
~ ;`-~
.,. ~
- 13 - :~
.. . . . . . . . . ..
3~
.._ _
~1
. ~ 0~ ID U~ ' C~
~ ~, U O O O ~i ,i C`J CD
~ .. . . _
.~
o i td ~ "~ I u~ c~
CO ~ N CD
._ _ _ .. _ _ . _ ._ __
~ O o O o O O g
~ CO ~ ~ ~
., bq . . _ ...... _ . ,___
,Q ~Oo ~ U~ C~
.` ~....... ~! C~ C~ U~ i
.. .. __ .__ _ . ,_ ___
.
~1 ~ ~ .
.. _ O p~ ._ ~ ... _
~1 ;c~l ~D_ _ CO_ .~ 11~ C~
.l _ ~1 C~ Ir) C~
~ ~ _ ~ _ __ .. _ --__ ___~__
~' l ~ O O O ' ~ C~ C~ ~
I _ ~ _ _~
~ P~ c~ ol~ ~
Uil o o . o, o o o o ~9
, _ _ .. . . _ ~
,: ~ ~,
¦ P~ ¦ I v c Is ~ .,1 1
~ .. _ . . . . , ~ q
~ , ~ ~ ~
~ ~ _E ~L~I
, . . . . . . . ........... . . . . . ... .... . . . . .. . . ...
.. . ; .. , .... ,`. .. .... ` . . .. . .. . .
OS3~L
The compounds thus show excellent
activity against both Gram- positive and Gram- negative
bacteria, especially against Gram- negative bacteria.
The compounds of formula (III) which
are used as starting materials in the process of the present
invention are also novel compounds, but these can easily be
prepared by known mlethods. Although they themselves have a
. .
- certain antibacterial activity, they are more valuable as
starting materials for the compounds of formula (II),which have
lO ~ much higher antibacterial activil;ies.
.. .
The invention is ~Lrther iLlustrated with
reference to the following Examples. The preparation of certain
novel starting m~Lterials is also illustrated in the following
~- Preparations 1 to 4.
, .
., .
,
:''; .
,:
- 15 -
:
5i3:~
Example 1
3Sarbamoyloxymethyl-7~-methoxy-7~ 2,4-triazol-3-yl-
thioacetamido) - 3 - cephem -4 -carboxylic acid
12 mg of 3-mercapto-l, 2,4-triazole were
~' 5 dissolved in 0.12 ml of a lN aqueous sol~ltion of sodium hydroxide;
to the resulting solution was added an aqueous solution of
sodium hydrogen carbonate containing 42 mg of 7,B-bromoacetamido-
3-carbamoyloxymethyl-7~-methoxy-3-oephem-4-carboxylic acid.
The mixture was agitated at room temperature for 2 hours, and
the pH was adjusted to about 2. 5 by addition of lN hydrochloric
acid. The mixture was then extrElcted with ethyl acetate, and
the organic phase was dried over anhydrous sodium sulphate.
The solvent was then distilled off under reduced pressure, yielding
4B mg of a crystalline residue. This residue was chromatographed
on a silica gel plate, employing a 50: 50 by volume mixture of
methanol and chloroform. The solvent was then distilled of E,
giving 33 mg of pure 3-carbamoyloxymethyl-7~-methoxy-7,B-(l,
2,4-triazol-3-yl-thioacetamido)-3-cephem-4-carboxylic acld.in
the form of a powder.
Nuclear magnetic resonance spectrum (CD3CN ~ D20), ô ppm:
5.00 (singlet, H at 6-position)
'
i ~.,
- 16 -
.
.
3~
3 . 90 ~singlet, -S- CH2-CO- )
3. 37 (singlet, OCH3 at 7 position).
Ultraviolet absorption spectrum (CH30H), ~ aX m~u:
263.
Infrared absorption spectrum (KBr), ~cm
1770.
Thin layer chromatography (silica gel~:
(a) Developing solvent (n- butanol/acetic
acid/water , 5: 4: 1 by volume) :
Rf value = 0 . 37 .
(b) Developimg solvent (methanol/chloroform,
:;
1 : 1 by volume):
Rf value = O . 28 .
~ ~ When this procedure was repeated except
; ~ 15 that the 7,B-bromoacetamido-3-carbamoyloxymethyl-7a!-methoxy-
3-cephem-4-carboxylic acid was replaced by the corresponding
7,8-chloroacetamido derivative, similar results were obtained.
~',:~
~ - Example 2
i~ 20 3-Carbamoyloxymeth~1-7,~ midazol-2-yl-thioacetamido)-7~-
methoxy-3-cephem-4-carboxylic acid
.~ ~
~ ,, , :
j
,
, '
!
~G~5~i3~
20 mg OI 2-mercaptoimidazole were
dissolved in 0. 2 ml of lN sodium hydroxide, and a solution of
117 mg of 7,B-bromoacetamido-3-carbamoyloxymethyl-7~-methoxy-
3-cephem-4-carboxylic acid in 0. 2ml OI lN sodium hydroxide was added
to the resulting solution, The mixture was a31owed to stand
at room temperature for 2 hours, after which the pH was
adjusted to 2. 5 by addition of 2N phosphoric acid. The mixture
- was then extracted with ethyl acetate and the aqueous phase was
freeze-dried and extracted with methanol. The methanol-
soluble portion was separated on a silica gel plate using a 50: 50
~ ~ by volume mixture of methanol and chloroform and extracted
-` with methanol. The solvent was distilled from the extract,
glving 80 mg of pure 3-carbamoyloxymethyl-7~3-(imidazol-2-yl-
i~ this~acetamido)-7~-methoxy-3-cephem-4-carboxylic acid in the form
;, 15 of a powder .
Nuclear magnetic resonance spectrum ~CD3CN f D20), ~ ppm:
7 . 09 (singlet, H of iIrlidazole)
5.0Z (singlet, H at 6-position)
3.75 (singlet, -S-CH2-CO-)
2~ 3,39 (singlet, OCE13 at 7-position).
- Ultraviolet absorption spectrum (phosphoric acid buffer, pH
' 6, 86)~ max ~u:
257 .
- 18 -
,
56~53~L
Infrared absorption spectrum (KBr)~cm
1765.
Thin layer chromatography (silica gel):
(a) Developing solvent (n-butanol/acetic acid/water,
5: 4: 1 by volume):
Rf value = 0. 21.
(b) Developing solvent (methanol/chloroform, 1 :1
.
- by volume):
.~ ~ ...
: Rf ~ 0. 31.
, . ~
, ......................................................................... .
When this procedure was repeated except
that the 7~B-bromoacetamido-3-carbamoyloxymethyl-7~-metho2~y-
3-cephem-4-carboxylic acid was replaced by the corresponding
7,B-chloroacetamido derivativeJ similar results were obtained.
:; :
Example 3
~ 3-Carbamoylo~ymethyl-7c~-methoxy-7,B-~lJ 3J4-thiadiazol-2-yl-
thioacetamldo)- 3 -cephem-4 -carboxylic acid
'':~ ' ~ ' :
A solution of 117 mg of 713-b~omoacetamido-:
3 -carbamoyloxymethyl- 7ar-methoxy- 3 -cephem-4 -carboxylic acid
in 0, 2 ml of lN sodium hydroxide was added to a solution of
: ' '
.' ''~ .'.
- 19 -
,. , ' . I
!
:~ , , . ,., , , ~ , . , : , - .,, -
i3~L
23.6 mg of 2-mercapto-1,3,4-thiadiazole in 0,2 ml of lN sodium
hydroxide, and the mixture was agitated at room temperature for
2 hours. The pE was then adjusted to 2. 5 by addition of 2N phosphoric
aeid. The solution was then extracted 5 times, each time with
15 ml of ethyl acetate, and the combined ethyl acetate extracts
were dried over sodium sulphate; the solvent was then distilled off,
giVillg 69 mg of an amorphous crude product. This was separated
.
on a silica gel plate using a 50: 50 by volume mixture of methanol
and chloroform and then extracted with methanol. The solvent
:, .
was distilled off from the extract, giving 34 mg of 3-carbamoyloxy-
methyl-7~-methoxy-7,B-(1, 3,4-thiadiazol-2-yl-thioaeetamido)-
:
3-eephem-4-carboxylic acid in the form of a powder.
` Nuclear magnetic resonance spectrum (CD3CM ~ D2O), ô ppm:
9. 30 (singlet, H of thlazole)
5. 03 (singlet, H at 6 -position)
.. ~ .
; 4,16 (singlet, S-CH2-CO-)
. 3.46 (singlet, OCH3 at 7-position),
Ultraviolet absorption spectrum (phosphorie acid buffer, pH 6. 86),
, ,
max m~
~6 3,
:
Imr ared absorption speetrum (~3r), ~ cm 1
1~60,
-20 -
.
.1 , 1
i3~
Thin layer chromatography (silica gel):
(a) Developing solvent (n-butanol/acetic acid/water,
5: 4 : 1 by volume):
Rf value = 0, 4 9,
-; 5 (b) Developing solvent (methanol/chloroform~ 1 :1
,;
by volume~:
Rf value = 0. 29 .
When thls procedure was repeated except
that the 7~-bromoacetamido-3-carbamoyloxymethyl-7cY-methoxy-
3-cephem-4-carboxylic acid was replaced by the corresponding
7,~-ch]oroacetamido derivative, similar result6 were obtained.
Examples of compounds prepared in
~ .
the ame rnanner as in the foregoing Examples and their properties
are shown below.
~ ' ' '
Except where otherwise indicated,the
physical properties were measured as follows:
Nuclear magnetic resonance spectrum (in CD3CN-~
; ~ D2)9 ~ ppm
Ultraviolet absorption spectrum ~in phosphoric acid
buffer of pH 6 . 86), ~max m~u.
.
21 -
;. ,, - , : . , . , ,- :
.
~56~3~
Infrared absorption spectrum (in KBr), ~J cm
D: Thin layer chromatography (TLC) on silica gel:
(a) Rf value obtained employing a 5: 4: 1 by volume
mixture of n-butanol/acetic acid/water.
(b) Rf value obtained employing a 1: 1 by volume
mixture of methanol/chloroform.
3-Carbamoyloxymethyl-7a!-methoxy-7~-(5-methyl-1, 3,4-thiadiazol-
2 -yl -thioacetamido) - 3 -c ephem -4 - carboxylic acid
NMR spectrum, ~ ppm:
5,02 (singlet, H at 6-position)
`, 4.05 (singlet, S-CH2CO-)
3.43 (singlet, OCH3 at 7-position)
2.64 (singlet, H of thiadiazole)~
IJV spectrum ~ max }~:
" . .
263.
IR spectrum~ cm
1770.
.. . . .
, TLC, Rf value:
(a)=0,45, (b)=0. 38,
3-Carbamoyloxymeth~yl-70~-methoxy-7,B-propargylthioacetamido-
3-cephem-4-carboxylic acid
~,' ' -
- 22 -
.
353~L
NMR spectrum ~ dimethylsulphoxide (~MSO)-d6], ~ ppm:
5.15 (singlet, H at 6-position~
3.40 (multiplet, -CH2-S~CH2 and OCH3 at 7-position),
UV spectrum.~ max ~lu:
2~2, 265.
IR spectrum~ cm 1
178~.
3~ arbamoyl xymethyl-7,B-cyanomethylthioacetamido-7a-methoxy-
3-cephem-4-carboxylic acid
NMR spectrum (DMSO-d6), ~ ppm:
5. 06 (singlet, H at 6 -position)
3, 58 (singlet, NC -CE2SCH2)
UV spectrum.~
246, 26 5,
l:R spectrum,~cm 1
~: 15 1720, 1776,
:. .
; 3-yl-thioacetamido)-3-cephem-4-carboxylic acid
~-~ NMR spectrum ~ ppm:
5.02 (singlet, H at 6-position)
~,05 (singlet, SCH2C~-~
23 -
.. ... . : . ~
~s~
3.43 (singlet, OCH3 at 7-position)
2 . 64 (CH3 of imidazole).
UV spectrum ;~ m~ m,u:
263,
IR spectrum~) cm
- 17~00
TLC, Rf value:
(a)= O. 39, (b)- O. 29.
. . . `
3-Carbamoyloxymethyl-7~3-(5-ethyl-1, 2,4-triazol-3-yl-thioacetamido)-
7cr-methoxy-3-cephem-4-carboxylic acid
NMR spectrum ~ ppm:
5,13 (singlet, H at 6-position)
- 4.00 (singlet, S-CH2-CO)
lS ~ ~ ~ 3. 52 (singlet, OCH3 at 7-position)
2. 83 and 1. 31 (quartet and triplet, CH3CH2 of triazole).
J: : `
UV spectrum . ~max mJl
, : . .
26 2.
,
, ~ IR spectrum,^Ycm 1 ~ ;
.j~ :
1765.
TLC, Rf value:
(a)=0.46, ~b)SO.33.
- 24 -
' .
i3~L
3-Carbamoyloxymethyl-7~-methoxy-7~ (5-propyl-1, 2,4-triazol-
3-yl-thioacetamido)-3-cephem-4-carboxylic acid
NMR æp ectrum 9 ~ ppm:
5.07 (singlet, H at 6-position)
3, 95 (singlet, S-CH2CO-)
3.49 ~singlet, OCH3 at 7-position).
IJV spectrum;l max m~
263.
IR spectrum,~ cm 1
176 5 .
TLC, Rf value;
(a)=0. 51, (b)=0.44.
3-Carbamoyloxymethyl-7a-metlloxy-7~-~5-phenyl-1, 2,4-tria~ol-
3-~1-thioacetamido)-3-cephem-4-carboxylic acid `~ ``
15~ NMR spectrum ~ ppm:
4 . 94 (singlet, H at 6 -position)
3. 92 (singlet, S-CH2-CO-)
,
:
3.38 (singlet, OCH3 at 7-position)
7.3 - 8.0 (H of benzene).
U~ spectrum~maX ~:
, 253, 235.
IR spectrum,~cm:
,....
- 25-
'
':
5~)53~
176 5 .
TLC, Rf value:
( a~=0 . 61, (b)0 . 53 .
3-Carbamoyloxy~ ethyl- 7~- methoxy-~3-(4-phenyl-1, 2, 4 -triazol-
3-yl-thioacetamido~-~cephem-4-carboxylic acid
acid
NM:R spectrum, ~ ppm:
4 . 99 ( singlet, H at 6 -position)
` 3.99 (singlet, SCH2CO-)
3~41 (singlet, OCH3 at 7-position).
- UV spectrum ~ max ~:
260.
. IR spectrum,~Jcm
7 6 5 .
., -
: TLC, Rf value:
(a)=0.43, (b)- 0. 38.
3 -Carbamoyloxymethyl- 7~-methoxy- 7,B - (4 -phenyl- 5 -methyl-l, 2, 4 -
tri~a~ol-3-yl-thi em-4-carboxylic acid
NMR spectrum, ô ppm:
;, :
5. 0V tsinglet, H at 6-position)
~ 3.92 (singlet, SCH2CO-)
,
3.42 (singlet, OC~I3 at 7-position)
~ - ,
'
. .
.
2.19 (singlet, CH3 OI trlazole).
UV spectrum, ~max ~:
260.
IR spectrum,)~cm l
1765,
TLCJ Rf value:
(a)=0 . 41, (b)=0 . 41.
... 3-Carbamo~methyl-7a-methoxy-7,B-(2-pyridylthioacetamido)- ;
3-cephem-4-carboxylic acid
NMR spectrum ~ ppm:
f , ,
5, 02 (singlet, H at 6 -position)
3.89 (~inglet, SCH2CO-)
3.39 (singlet, OCEI3 at 7-position)
. . 7, 0 - 8. 5 (H of pyridi~le).
UVspectrum,~ m,~:
266, 239.
IR spectrumy cm l
1770.
TLC~R val~le: ~ ` . .
f
(a)=0. 57, (b)=0.43,
3-Carbamoylox~rm thyl-7~Y-methox~-7f3-(4-met lyl-2-pyrimi~lthio-
. ~ acetamido)-3-cephe~ 4-carboxylic acid
'f. ~'
f .
.
- 27- :
5~
.
NMR spectrum, ~ ppm:
5 . 02 (singlet, H at 6 -position)
3. 93 (singlet, SCH2CO-)
~; 3.43 (singlet, OCH3 at 7-position)
2.42 (singlet, CH3 of pyrimidine).
UV spectrum. ~ maX m~U:
242, 270.
IR spectrum,~cm :`
1780.
.1 :
TLC,Rf value:
(a~=0. 51, (b)-b. 39.
3-Car amoyloxymethyl-7~-methox;~7~imidylthioacetamido~-
;` 3-cephem-4-carboxylic acid
.
NMR spectlr~li~, ô pprn:
.
15 ~ 5.04 (singlet, H at 6^position)
3.96 (singlet, SCH2CO-)
'~ - 3.46 (singlet, OCH3 at 7-position)
7. 22 and 8, 58 (triplet and douhlet, H of pyrimidine).
UV spectrum,~maX rryu:
,' 20 242, 26~.
.
IR spectrum, ~ cm 1
1760, 1700.
- 28 -
;'' ~ ' .
, ............................................................. .
TLC, Rf value:
(a)=0,43, (b)=0.43.
3-Carbamoyloxymethyl-7~-methoxy-7~-(8-purinylthioacetamido)-
3-cephem-4-carboxylic acid
NMR spectrum, ~ ppm:
4. 99 (singlet, H at 6-position)
3 . 37 (singlet, OCH3 at 7 -position)
8.65 and 8.70 (H of purine).
UV spectrum,~maX mp
- 10 2~7.
IR spectrum,~cm
1770.
, TLC, Rf value:
(a)=0. 35, (b~=0.18.
~- 15 3-Carbamoyloxymethyl-7~-methoxy-7~-(6-purlnylthioacetamido)-
3-cephem-4-carboxylic acid
NMR spectrum, i~ ppm:
5.01 (singlet, H at 6-position)
4.14 (singlet, SCH2CO-)
2û 3.43 (singlet, OCH3 at 7-position)
8. 31 and 8. 63 (E of purine) .
.
.
- 29 - L
.
. , . . , ~ , ~ , . .
3~.
UV spectrum~maX m~
278.
IR spectrum,V cm:
176 5.
TLC, Rf value:
(a)=0.40~ (b) 0. 20.
7,B-(2-Benzimidazolylthioacetamido)-3-carbamoyloxymethyl-7ar-
- methoxy-3-_ephem-4-carboxylic acid
~ . .
NMR spectrumJ ~ ppm:
.,
4 . 95 (singlet, :H at 6 -position)
4.16 (singlet, SCH2CO-)
3.41 tsingletJ OCH3 at 7-position).
UV spectrum~ma ~:
2~1, 287.
IR spectrum, ~ cm : -
1780, 1680.
~ TLC, Rf value:
.; - (a)=0.48, ~b)=0. 51.
-
7~3-(2 1S~A~tamido)-3-carba~noyloxymethyl-7~-
metho~ ~em-4-carboxylic acid
, ~
~ ~ NMR spectrum, ~ ppm:
- .
., .
:, .
,
~, ~ ....... . . . . . . .
~5~i3~L
5 . 05 (singlet, H at 6 -position)
- 4.14 (singlet, SCH2CO-)
3. 50 (singlet, OCH3 at 7-position).
UV spectrumJ ;~ max m~
283, 276J 245.
IR spectrum ~ cm : .
1~80J 172~.
~; TLC J Rf value:
. (a)=0, 61J (b)=0.48.
: 10 7~ - ( 2 -B enzthla7.olylthioac etamido) - 3 - carbamoyloxymethyl - 7~-
methoxy-3-cephem-4-carboxylic acid
NMR spectrum (CD3OD)J ~ ppm:
5. 04 (singletJ H at 6 -position)
3,45 (singletJ OCH3 at 7-position),
: lS UV spectrumJ~ maX
: : 2~7, ~71.
,j~: - ,
IR spectrumJ ~ cm
1770.
TLC, Rf value:
ao (a)-0 . 58 J (b) =0 . 48,
.: , ' ,
- 31-
.
53~L
3 - Carbamo~oxymethyl- 7~-m ethoxy- 7,e - ( s -tria zolo~ 4, 3 -a] pyridin-
3 -yl-thioac etamido) - 3 - cephem -4 -carboxylic acid
NMR spectrum, ~ ppm:
4. 94 (singlet, H at 6-position)
3.95 (singlet, SCH~CO-)
3.40 tsinglet, O(:H3 at 7-position~.
UV ~;pectrum,~maxrr~:
269.
IR spectrum,~cm l
1770, 1690.
TL(, R value:
(a)=0. 29, (b)=0. 39.
,: :
3-Carbamoyloxymethyl-~-methoxy-7~-t2-naphth~hioacetamido)-
- ~ 3-cephem-4-carboxylic acid
":
l~ ~ NMR spectrum, ~ ppm:
-
4. 92 (singlet, H at 6-position)
3.72 (singlet, SCH2-CO-)
3.29 (singlet, OCH3 at 7-position)
7. 3 - 7. 9 (H of naphthalene).
., .
(~ UV spectrum,~ ma~
248, 270 (Sh.).
.
- 32 -
, ,
S3~
IR sp ectrum, ~ cm
176 5.
TLC, Rf ualue:
(a)=0. 61, (b)=0. 60.
~ "Sh"= "shoulder"]
3-Carbamoyloxymethyl-7a-methoxy-7~-(2-thiazolinylthioacetamido~-
: ~ 3 - cephem -4 - carboxylic acid
:' " i
NMR sp ec1; rum 9 ~ ppm:
5. 08 (singlet, H at 6-position)
~ - 10 3, 92 ~singlet, SCH2CO-)
~ .
! 3. 50 (singlet, OCH3 at 7-position).
UV spectrurn,~max m}
258, 239.
IR spectrum )~cm 1;
S~ 1770, 1690.
TLC, Rf ~ralue:
. (a)=0.49, ~b)=0.49,
~ , .
3-Carbamoyloxymethyl-7~-cyclohexylthioacetamido-7~-methoxy-
3-cephem-4-carboxylic acid
.;
:~ Z0 NMR spectrum, ~ ppm:
5. 08 (singlet, H at 6-positlon)
3 . 81 (einglet, SCH2CO-)
,, .
33 --
, , .
" ~'.'
3~
3.48 (æinglet, OCH3 at 7-position)
1. O - 2. (CH2 f cyclohexane) .
UV spectrum,~ a ~:
266.
TLC, Rf value:
(a)=O, 56, (b)=O. 50.
... 3-C:arbamoyloxymethyl-7~-methoxy-7,B-(rl-~ropylthioacetamido)
3-ceRhem-4-carboxylic acid
~ ' , .
N~R spectrum (CD30D), ~ ppm:
5, 04 (singlet, H at b-position)
3.45 (singlet, OCH,~ at 7-position3.
UV spectrum,~maX rryu;
26 5.
.~ ~ IR spectrum,~cm 1
': : . 1771~.
.,
TLC, R~ value:
`` (a)=O. 55, (b)=O. 55.
3-Carbamo;sr~oxymethyl-q~-methoxy-7~,_-pent2~ioacetamido)-
3-cel?hem-4~ ~ d
,. ~ .
. ~ .
:? :
.' - . , '
' . ,. , , .. , .,, .. : : ' .
i3~
NMR spectrum (CD30D), ~ ppm:
5, 04 (singlet, El at 6 -position)
3.46 (singlet, OCH3 at 7-position).
UV spectrum. ~ max rryu:
265.
IR spectrum,~)cm 1
1770.
TLC, Rf value:
(a)=O, 60, (b)=O. 54,
';i . ,
- 10 Example 4
:i
~;~ 7~B-(2-Butylthioacetamido)-3-carbamoyloxyrreth~rl-7~-methoxy- 3-cel?hem-4-carboxylic acid
45 mg of sec-butanethiol were dissolved with 0. 5 ml
of a lN aqueous solution of sodium hydroxide in 50% aqueous
methanol, and 0. 5 ml of benzhydryl 7,B-bromoacetamido-3-
carbamoyloxymethyl-7~-methoxy-3-cephem-4-carboxylate ~ras
added to the resulting solution, The mixture was agitated for
2 hours, after which methanol was distilled off under reduced
pressure. The aqueous phase waæ extracted three times with
~; 20 ethyl acetate and the extracts were dried over sodium sulphate.
The solvent was then distilled off, giving crude benz~ydryl 7~-
,' .
- 35 - ~
' ' , .
.
~ 533L
(2-butylthioacetamido) - 3 - carbamoyloxym ethyl- 7~-methoxy- 3 -
cephem-4-carboxylate. This crude product was dissolved in
1. 0 ml of anisole, and 1, 5 ml of trifluoroacetic acid were added
to the resulting solution. The mixture was allowed to stand at
room temperature for 6 minutes and then evaporated to dryness
under reduced pressure. The solid residue was dissolved in
20 ml of ethyl acetate and 20 ml of 0. 2M phosphoric acid buffer
(pH 7. 5) and the mixture was agitated and then allowed to stand
to allow phase separation. The aqueous phase was washed once
again with ethyl acetate and its pH was then adjusted to 2. 5. The
aqueous layer was then extracted 4 times with 20 ml of ethyl
acetate, and the combined extracts were dried over sodium
sulphate and evaporated to dryness under reduced pressure giving
an amorphous crude product, This product was separated and
purified on a silica gel plate emplo~ying a 50 : 50 mixture of
methanol and chloroform. 115 mg of the desired product were
obtained in the form of a powder.
~, Nuclear magnetic resonance spectrum (CD30D), ~ ppm:
5.08 5singlet, H at 6-position~
.
3. 58 (singlet, OC~13 at 7-position).
Ultraviolet absorption spectrum (CH30H) 1 maX ~:
- ` 26~ (= 9i40).
.
~ - 36- -:
'
~ . . . . . . .. . . . . . . . . . .. .
S(~S3~L
Ihrared absorption spectrum (KBr), Y cm 1
1795, 1720, 1680.
Thin layer chromatography (silica gel):
(a) Developing solvent (n-butanol /acetic acid/water, -
5: 4: 1 by volume):
` :Rf value = 0. 59.
~` (b~ Developing solvent (chloroform/methanol,
1 ~ 1 by volume):
Rf value = 0 . 6 0 .
. . .
`; 10 ~5
3-Acetoxymethyl-7~-methoxy-7~ 3, 4-thiadiazol-2-yl-thioacetamido)-
3-cephem-4-carboxylic acid
. .
~. . 33 mg of 2-mercapto-1, 3, 4-thiadia~.ole were di.~solved
.~: .
: in 0. 24 ml of lN aqueous sodium hydroxide, and a solution of
~: 15 0.27 mmole ~100 mg) of 3-acetoxyrnethyl-7~-chloroacetamido-7~- :
methoxy-3-cephem-4-carboxylic acid m 0, 5N sodium bicarbonate
: was added to the resulting solution. The mixture was agitated :.
at room temperature for 3 hours and the p~I was then adjusted
.
to 2. 5 by addition of O.lN phosphoric acid. The mixt7lre was then
extracted 3 times with ethyl acetate and the combined ethyl
- . acetate extracts were dried over sodium sulphate. The solvent
.,'" ~"
.-'
- 37 -
.
- ~S~53~
.
was distilled off from the dried ethyl acetate solution, giving
113 mg of an arnorphous crude product. This was separated on a
silica gel plate employing chloroform containing 40% methanol
as a solvent and was then extracted with methanol. The solvent
vvas distilled off from the extract, giving 69 mg of the desired
- ~ product .
.,~ .
Nuclear magnetic resonance spectrum (CD3CN -~ D2O), ~ ppm:
9 . 23 (singlet, H of thiadiazole)
5. 00 (singlet, H at 6-position)
4.73 (doublet, CH2 at 3-position)
~r 4.11 (smglet, S-CH2-CO-)
~' 3.42 (singlet, OCH3 at 7-position)
1. 98 (singlet, O-CO-CH3).
IJltrav~olet absorption spectrum ~phosphoric acid buffer of pH 6, ~6j,
; ~ ~ lS ~ max P
26 3 m~ =87 89) .
Infrar ed absorption spectrum (KBr), ~ cm 1
1760.
.:: .
Thin layer chromatography (silica gel):
~ Developing solvent (methanol/chloroform, 1: 1 by volume):
R value = 0.45.
f
Examples of other compounds prepared
.
in the same manner as in the foregoing Example, together with
.
- 3 8
' , , ' ' . ..
,
, , , . . .:
`, : : ' , ~ ! `
~L~S~53~L
their properties (determined as described in Example 3) are
shown below:
3 -Ac etoxym ethyl - 7~ - cyanom ethylthioac etamido - 7a -
methoxy-3-cephem-4-carboxylic acid
NMR spectrum (CD3CN), ~ ppm:
5 . 06 (singlet, H at 6 -position)
4.76 - 5.06 (quartet, -CH2OCO- at 3-position~
3,60 (singlet, NCCH2S or SCH2CO)
3. 52 (singlet, OCH3 at 7-position)
3,42 ~singlet, NCCH2S or SCH2CO)
3, 32 - 3, 55 (quartet, H2 at 2-position)
2,02 (singlet, OCOCH3).
UV spectrum ~
247 ( ~ = 8000)
267 ( '~ -8400).
IR spectrum,2J cm 1
177 5,
3-Acetoxymethyl-7~-methoxy-?~B-pro~argylthioacetamiào-
3-cephem-4-carboxylic acid
N~R spectrum ~DMSO-d6j, ~ ppm:
5.05 (singlet, H at 6-position)
4.9 - 4.6 (quartet, -CH2OCO- at 3-position)
i3~
3. 36 (singlet, OCH3 at 7-position)
3. 2 - 3. 5 (multiplet, 2-position, H2, -C:H2 and
-S-C~2 )
3.05 (triplet, HC_C)
1.98 (singlet, OCOCH3).
UV spectrum,;~max ~yU:
245 (~ =7800)
.
~... 268 (~ =8200).
IR spectrum,lJ cm 1
1775.
When 83 mg of the carboxylic acid
thus obtained were added to 8 m]. of water, 17 mg of sodium
bicarbonate were gradually added thereto and the resulting
homogeneous solution was freeze-dried, a sodium salt of tlle
carboxylic acid was obtained in the form of an amorphous
powder.
, 3-Acetoxymethyl-7,B-azidomethylthioacetamido 7~-methox~sr-
3-cephem-4-carboxylic acid
P~MR spectrum (DMSO-d6), ~ ppm:
5.14 (singlet, H at 6-position)
4.69 - 4.98 (quartet, -CH2-OCO- at 3-position)
- 40 -
'
3~. -
4. 51 (singlet, N3CH2S-)
3.40 (singlet, 7-position, OCH3, S-CH2-CO)
3.3 - 3.6 (quartet, H2 at 2-position)
2 00 (singlet, OCO-CH3).
5 UV spectrum,~max m~:
` 247 ( =57800)
269 ( = 8000).
IR spectrum, 1J cm 1
1775.
. 10 3 -Acetoxymethyl-7~3- (imidazol-2-yl-thioacetamido) - 7a-methoxy-
3-cephem-4-carboxylic acid
N.MR spectrum ~ ppm:
7.11 (singlet, H at 4- an~d 5-position of imidazole)
5 . 0 2 tsinglet, H at 6 -position~
4.78 (broad singlet, H2 at 3-position)
3.78 (singlet, -S-CH2CO-)
3 38 (singlet, OCH3 at 7-position)
2.02 (singlet, -O-COCH3).
UV spectrUm,,J~ max
265.
IR spectrum, 2~ cm
. 1760.
TLC, Rf ~alue:
(b) =0. 46 .
_ 41 -
~ ' .
531
Example 6
7a-Methoxy- 3 - (l-m ethyl -lH-tetrazol- 5 -yl)thiomethyl- 7~3-(1, 3, 4 -
thiadiazol-2-yl-thioacetamido)-3-ceE~hem-4-carboxylic acid
33 mg of 2-mercapto-1, 3, 4 -thiadiazole were dissolved
in 0. 24 ml of lN sodium hydroxide and a solution of 117 mg
(0.27 mmole) of 7,B-chloroacetamido-7~-methoxy-3-(1-methyl-
lH tetrazol- 5-yl)thiomethyl- 3 -cephem-4 -carboxylic acid in
0. 5N sodium hydrogen carbonate was added to the resulting
solution. The mixture was agitated at room temperature for
3 hours, and the pH was then adjusted to 2. 5 by addition of
0,lN phosphoric acid. The mixture was extracted 3 times
with ethyl acetate, and the combined extracts were dried over
sodium sulph~te. The solvent was then distilled off, giving 125 mg
OI an amorphous product. This was separated on a silica gel
plate employing a 50: 50 mixture of methanol and chloroform
and extracted with methanol. The solvent was distillèd off from
the extract, giving 70 mg of the desired product in the form of
, an amorphous powder.
Nuclear magnetic resonance spectrum (CD3CN + D2O), ~ ppm:
ZO 9, 28 (singlet, H at 5-position of thiadiaæole).
Ultraviolet absorption spectrum (phosphoric acid buffer of pH
6 . 86 ) ~ max m~:
266 (~ ~ 9270).
- 42 -
~'' ' , . ' . ~
3~L
; Infrared absorption spectrum (K~r), ~Jcm
. 176 0 .
Thin layer chromatography ~ silica gel):
I)eveloping solvent (chloroform/methanol, 1:1 by volume)
Rf value ~ 0. 38.
:''
Examples of compounds prepared in the same manner
as in the foregoing Example and their properties (measured
as described in Example 3) are shown below:
"
7,8-(Imidazol-2-ylthioacetamido)-7a!-methoxy-3-(1-methyl-lH-
. .
~ :l0 tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid
:
NMR spectrum ~ ppm:
`~ 7.03 (singlet, H at 4- and 5-position of imidazole~
: 4. 95 ~singlet, H at 6-position)
3 . 84 (singlet, CH3 at l-posltion of tetrazole ring)
: 15 3. 36 (singlet, OCH3 at 7-positionj .
1 . :
UV spectrum, ~max
: 265 (~= 7200).
- IR spectrum, ~J cm l
.~ 1760.
TLC, Rf value:
(b) = 0 . 36 .
: :
~ ~ -43
, . . . .
,. .. . . . . .
~CI Sq~153~il
7,B=cyanomethylthioacetamido-7~methoxy-3-(l-methyl-lH
tetra zol - 5 - yl)thiom ethyl - 3 - c eph em - 4 - c arboxylic acid
NMR spectrum, ~ ppm:
5.10 (singlet, EI at 6-position)
4.3 - 4.6 (quartet, CH2-S at 3-position)
3. 98 (singlet, CH3 at l-position of tetrazole ring)
3 . 70 (singlet, -NCCH2S or -SCH2CO)
3.5 - 3.7 (quartet, H2 at 2-position~
3.60 (singlet, -NCCH2S or -SCH2CO)
3 . 50 ¦sin~let, OCEI3 at 7 -position) .
` UV spectrum ~ mp:
274 ( = 9ooo),
7a-~$ethoxy- 3 - (l-methyl -lH-tetrazol- 5 -yl)thiomethyl- 7,~ -propargyl-
thioacetamido-3-cephem-4-carboxylic acid
NMR spectrum (DMSO-d6), ~ ppm:
. ~ ~.05 (singlet, EI at 6-position)
4 . 2 - 4 . 3 (quartet, CH2-S at 3 -position)
3. 90 (singlet, CH3 at l-position of tetrazole ring)
about 3. 5 (multiplet, OCH3 at 7-position, H2,
-CEI2-S-CH2CO at 2-position)
3,20 (triplet, HC_C-).
- 44 -
.
.
, , , . ~ . : .
1~5~53~
Example 7
7~-Methoxy-3-(1-methyl-lH-tetrazol-5-yl )thiomethyl-7,B-(1, 3,4-
_
thiadiazol- 2-yl-thioacetamido)- 3-cephem-4 -carboxylic acid
To a solution of 596 mg of 2-[ 7,B-bromoacetamido-
7~-methoxy-3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-
:~ 4-carbonyl] -s-triazolo[ 4,3-a] pyrid-3-one in 30 ml of acetone
was added a solution of 86 mg of 2-mercapto-1, 3,4-thiadiazole
and 84 mg of sodium hydrogen carbonate in 8 ml of water; the
resulting mixture was stirred at room temperature for 1 hour,
-1 lû The solvent was then distilled off under reduced pressure and 50
~ ml of ethyl acetate were added to the residue, which was then washed
~ with water and dried over anhydrous magnesium sulphate. The
solvent was distilled o~, giving crude 2 [7a-metho~y-3-(1-methyl-lH-
tetrazol-5-yl)thiomethyl 7~3-(1, 3,4-thiadiazol-2-yl-thioacetamido)-
3 - cephem-~ -carbonyl~ - s - triazolo[ 4, 3 - a] pyrid- 3 -one as a powder .
Nuclear magnetic resonance spectrum (CDC13), ~ ppm:
3.43 (singlet, O-CH3 at 7-position)
3. ~0 (singlet, N-CH3 at 3-position of tetrazole)
4, 25 (singletJ -S-CH2-CO at 7-position)
. 5,24 (singlet, 6-position, ~
--N~ H
'~ 6,3 - 7~9 (multlplet, 4-position -CO-N~
O H
.' ' ' .
,
:
.
. .
5~3~
9,08 (7-position, thiadiazole)
Ultraviolet absorption spectrum (THF), max nm:
227, 261, 359.
Infrared absorption spectrum (Nujol-trade mark), L~cm l
1770, 1700, 1650,
Thin layer chromatography ( silica gel):
(a) Developing solvent (chloroform containing 10%
. methanol~:
' R E value = O . 56 .
; 10 (b) Developing solv~ent (n-butanol/acetic acid/water,
. 4: 1 :1 by volunne): ;
Rf value - 0 . 47 .
To a ~olution of the compound obta~ned above in a
mi~ture of 15 ml of tetrahydrofuran and 7. 5 ml of water were
.
- . 15 added 600 mg of copper acetate monohydrate. The resulting
~' ~ mixture was stirred at room "emperature for 5 hours, after
, .
which 50 ml of ethyl acetate and 7. 5 ml OI a 0. 5M citric acid
, .
solution were added thereto. Insolubles were filtered off
.'
and the organic phase was washed three times with 50 ml OI
an aqueous solution of sodium chloride and then dried over
anhydrous magneslum sulphate, The solvent was distilled off~
under reduced pressure to give a crude amorphous product,
:. . ' .
~'
,: ~
., .. ' '
53~
which was purified by thin layer chromatography, using as
solvent a mixture of n-butanol/acetic acid/water (4: 1 :1 by
volume), to give the desired product.
Nuclear rnagnet;c resonance spectrum (CD3CN + D20), ~ ppm:
3.46 (singlet, O-CH3 at 7-position)
3.94 (singlet, 3-position, tetrazole N--CH3)
6.02 (singlet, 6-position, Hy
. ~ . ~N~
.. ..
9,28 (singlet, 7-position, ~J~S3~ )
Infrared absorption spectrum (KBr) 1~ cm 1:
:, 176 0 .
~, Ultraviolet absorption spectrurn (in phosphoric acid buffer of
: ! pH 6, 86)~max nm
; 2~6,
' : Thin layer chromatography (~ilica gel):
eveloping solvent (chloroform/methanol,l :1 by
J ~ volume):
Rf value = 0, 38,
'
~8
i 7,B (2-Carbo~y4henylthioacetamido)-7~-methoxy-3-(1-methyl-lH-
tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid
, ~ '.
- 4 7
. .
3~a
To a solution of 596 mg of 2-[ 7,B-bromoacetamido-
7~-methoxy- 3 - (1-methyl -lH -tetrazol- 5 -yl)thiomethyl- 3 -cephem -4 -
carbonyl] -s-triazolo[ 4, 3-a] pyrid-3-one in 30 ml of acetone was
added a solution OI 152 mg of thiosalicylic acid and 173 mg of
' 5 sodium bicarbonate in 8 ml of water. The resulting mixture was
stirred at room temperature for 30 minutes, The solvent was
` then distilled off under reduced pressure and 50 ml of ethyl
' acetate were added to the residue. The resulting solution was
then washed with water and dried over anhydrous magnesium
æulphate. The solvent was then distilled off under reduced pressure,
giving crude 2-[ 7,B-(2-carboxyphenylthioacetamidoj-7,x-methoxy-
, :
3-(1-methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carbonyl] -s-tria~olo-
1 4, 3-a] pyrid-3-one as a powdery residue,
. . .
, This product was dissolved in a mixture of 15 ml oi
tetrahydrofuran and 7. 5 ml of water and then 500 mg of copper
,,
'~ ~ acetate monohydrate were added thereto. The resulting mixture
~ : . :
',, was stirred at room temperature for 5 hours, after which 50 ml
,~ .
of ethyl acetate and 7~ 5 ml of a 0. 5M citric acid solution were
!~- added thereto. I~isolubles were filtered off and the organic phase
2~ was was,hed three times with an aqueous solution of sodium
.
~` ~ chloride and then dried over anhydrous magnesium sulphate.
:
:, ' '
,, - 48 -
:;
.
. .
~5~3~
~; After the solvent had been distilled off under reduced pressure,
the residue was purified by thin layer chromatography on silica
gel, using as solvent a mixture OI n-butanol/acetic acid/water
(4: 1: 1 by volume), to give the desired product.
Nuclear magnetic resonance spectrum (CD3CN ~ D20), ô ppm:
- 7. 2 - 7.6 (multiplet, proton in phenyl moiety)
5.13 (singlet, 6-position, ~~ )
~N~
.:. ~ ' i
4.07 (singlet, 3-position tetrazole ,,N-CH3)
3.47 (singlet, 7-position, -O-C~H3),
rared absorption spectrum (KBr) ~Jcm 1
1775.
.
Ultraviolet absorption spectrum (phosphoric acid buffer, pH
6. 86)J A nm:
: 15 253
~'' .
~` Thin layer chromatography (silica gel):
.
- (a) Developing solvent (n-butanol/acetic acid/
water , 4 :1 :1 by volume) :
;: : Rf value = 0. 36.
(~) Developing solvent (chlorolorm/methanol/
water, 6: 4: 1 by volume):
'- Rf value = 0. 20.
,
. 49
. , .
... . .
Example 9
7,,~ - ( 2 - H~droxyethylthioac etamido) - 7~- m ethoxy- 3 - (1- m ethyl -lH -
tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid
434 mg of 7,~-chloroacetamido-7~-methoxy-3-(1-
methyl-lH-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid
were ~igsolved in 20 ml of water containing 84 mg of sodium
` bicarbonate and 156 mg of thioglycol were added thereto. The
- ~ resulting mixture was then stirred at room temperature for 3 hours,
while maintaining the pH at 8. 0-3. 5 by addition of a lN sodium hydroxide
solution, The pH was then adjuslted to 2. 0 - 2, 5 by addition of lN
hydrochloric acid and the mi~ture was freeze-dried. The residue
1: was extracted with methanol and the olvent was distilled off. The
resulting residue was then purified by preparative silica gel
chromatography u~ing as developing solvent a mixture of n-butanolJ
15 ~ acetic acid/water (4 :1: 1 by volume)9 to give 360 mg of the
desired product as a powder.
: i
I` Nuclear magnetic resonance spectrum (CD3CN + D2)J ~ ppm:
5.2G (singlet, 6-position,
.i
,N~
4,15 - 4.25 (quartet, 3-position, \1CH2-S-)
'' ' .
3.90 (~inglet, 3-position, tetr~zole , ~ )
CH3
- 50 -
, ' .
.
533~
3.44 (singlet, 7-position, -O-CH3)
3. 30 (singlet, 7-position, -S-CH2-C-)
3.68 and 2.74 (triplet, 7-position, HO-CH2CH2-S-).
Infrared absorption spectrum (KBr), lJcm 1
1740, 1675.
. .
Ultra~iolet absorption spectrum (phosphoric acid buffer, pH
6. ~6)~ ~max m~:
270 ( = 9450)
Preparation 1
7,B-Bromoacetamido-3-carbamoyloxymethyl-7~-methoxy-3-
~'~ 10
cephem-4-carboxylic acid
Into a beaker were charged 808 mg of
bromoacetyl bromide and 1.030 g of bis-(trimethylsil~)triElworoacet~ id~
~-~ and the resulting m~xture was allowed to stand at room
, :
temperature for 20 minutes. 5 ml of methylene chloride
were then added to the mixture, followed by a solution of 879 mg
of dibenzhydryl 7,B-(D-5-t-butoxycarbonylarnino-5-carboxyvaler-
amido) - 3 -carbamoyloxymethyl- 7~-methoxy- 3-cephem-4 -carboxylate
in 5 ml of methylene chloride. Thebeaker was washed with
10 ml of methylene chloride and the washings were combined
,
with the mixture. The mixture was aLlowed to stand at
. .i
room temperature under moist conditions for 2 hours. 1 g
of sodium bicarbonate was then added to the reaction mixture,
:~ .
- 51 -
whilst cooling with ice/sodium chloride, and then 20 ml of
a 5% aqueous solution of sodium bicarbonate was added to
the reaction mixture, which was agitated for 30 minutes.
The methylene chloride phase was combined with the washings
obtained by washing the water phase wlth 10 ml of methylene
chloride, and the mixture was washed twice with 20 ml of a
20% aqueous solution of sodium chloride. The organic phase
was dried over sodium sulphate and the solvent was distilled
off under reduced pressure, giving 1.194 g of crude diben~hydryl
7,B-[ (D- 5-t -butoxycarbonylamino- 5-carboxyvaleryl)-bromo-
ac etylamino~ - 3 - carbamoyloxym ethyl - 7a - m ethoxy- 3 - c ephem -
~, 4-carboxylate in the form of a yellow amorphous product.
, .
This ye:Uow pr~duct was dissolved in 1 rnl
~; of anisole and 2 ml of trifluoroacetic acid, and the resulting
' .
~ solution was allowed to stand at room temperature for 5 minutes,
;
after which the solvent was distilled off under reduced pressure.
, .
The r esulting yellow viscous paste was dissolved in 20 ml
.,j
' of ethyl acetate and 20 ml of a 0. 2M phosphoric acid buffer
~pH 7~ 5), and the solutlon was then transferred into a separating
funnel and well shaken. After phase separation, the pH of
the aqueous phase was adjusted to 2. 5 with lN hydrochloric
. '
, .
~. -52 -
,. .
~ .
~ L~5~3~3~
acid and the aqueous phase was extracted 5 times with 20 ml
of ethyl acetate, The extracts were dried over sodium sulphate,
after which the solvent was distilled off under reduced
~`
pressure. 300 mg of the desired compound were obtained
S ~n the`form of a crude amorphous product.
Nuclear magnetic resonance spectrum (CD3CN ~ D20), ~ ppmL
5.12 (singlet, H at 6-position)
3.95 (singlet, BrCH2CO)
~, . . .
3. 53 (singlet, OCH3 at 7-posltion),
IJltraviolet absorptlon spectrum (CH30H), lmaX, mJl:
263.
Infrared absorption spectrum (KBr),~cm
1780, 1700.
Thin layer chromatography (silica gel):
(a~ Developing ~olvent (n-butanol/acetic acid/
.
water, 5: 4 :1 by volume~:
Rf value O. 53.
' .
(b) Developing solvent (methanollchloroform, 1:1
by volume):
Rf value - 0~44.
Preparation 2
3 -Carbamoyloxymethyl- 7~3- chloroacetamido- 7a-methox~3-
., , ~
ce,phem-4-carboxylic acid
- 5 3 -
~ .
- ~S~Si3~
The proeedure described in Preparation 1 was
repeated except that chloroacetyl ehloride was employed instead
of the bromoacetyl bromideO Separation and puriiication took
place as described in Preparation 1, giving the desired eompound
.
in the form of a crude amorphous produet.
Nuelear magnetie resonanee speetrum ~CD3CN + D2O), ~ ppm:
5.1 (smglet, H at 6-position)
3.48 (singlet, OCH3 at 7-position)
.
-~ 4.11 (singlet, ClCH2CO).
Ultraviolet absorption speetrurn (phosphorie aeid buffer, pH 6.86),
max, m~
~6D~.
i. Infrared absorption speetrum (KBr), ~cm 1
~ 1780, 1700,
:: 15 Thin layer chromatography (silica gel):
..
, : Developing solvent ~methanol/ehloroiorm, 1 :1 by
~ . .
- volume):
Rf value = 0 . 35 .
~ ~:
PreE~aration 3
`
3-~Aeetoxymethyl-7,l3-ehloroacetamido-7~-methoxy-3-eephem-
: :
~38 . 5 mg oi benzhydryl 3-aeetoxymethyl-7~e-
ehloroaeetamido-7a~-metho~y-3-eephem-4-earbox~rlate were
. ' .
.,
' , ,
.; , .. , .. . . . . . , . , . : . . :. :
~5~53~
dissolved in 0.4 ml of anisole, and 0.8 ml of trifluoroacetic acid
was added to the resulting solution. The mixture was agitated at
room temperature for 5 minutes. The reaction mi~ture was then
prompMy evaporated to dryness and the residue was washed with
n-hexane and dissolved in 5 ml of 0. 25M phosphoric acid buffer
(pH 7. 5). The resulting solution was neutralized with a 5% aqueous
solution of sodium bicarbonate and washed with ethyl acetate. The
pH of the aqueous phase was adjusted to 2. 0 with 60% phosphoric
acid and the oily substance which precipitated was extracted with
ethyl acetate. The extract was washed with water and dried over
anhydrous sodium sulphate, after which the solvent was disti~led
off, giving 276 mg of the desired compound in the form of an
- ~ ,
amorphous powder.
Nuclear mag,netic resonance spectrum (CD3(:N -~ D20), ~ ppm:
;~ 15 5.08 (singlet, H at 6-position)
4.08 (singlet, S:lCH2CO)
3.55 (singlet, OCE3 at 7-position).
Thin layer chromatography (silica gel):
Developing solvent (chloroform/methanol, 9 :1 by
20 ~ volume):
1 ~ -
,~ ' -
'
53:~
Preparation 4
~,B-Chloroacetamido-7O~-methoxy-3-(1-methyl-1 -tetrazol-5
yl)thiomethyl-3-cephern-4-carboxylic acid
27.6 g of disodium 7,B-(D-5-amino-5-carboxyvaler-
aimido) - 3 - carbamoyloxym ethyl- 7a -m ethoxy - 3 - c ephem - 4 - carboxylate
were dissolved in 1090 ml of a 5% aqueous solution of dipotassium
phosphate, and 715 ml of acetone were added to the resulting
solution. 8.1 g OI 4-dimethylaminopyridine were then added to
the solution, whose pH was adjusted to 9. 5 by addition of
2, 5N aqueous sodium hydroxide, following which 34. 5 ml of
t-butoxycarbonyl azide were added and the mixture was agitated
at room temperature for 4 hours while maintaiining the pH at 9. 0
to 9.5. The reaction mixture was allowed to stand at 4C. overnight,
at the end of which time 1000 ml of ethyl acetate were added
;' 15 thereto, and the mixture was well shaken. The aqueous phase
' ~ was collected and another 1000 rnl of ethyl acetate were ~dded
thereto, after which tbie PE of the aqueous phase was adjusted
to 2, 5 by addition of concentrated hydrochloric acidJ while
maintaining the temperature at 0 to 2 C, The organic phase
. ~ .
was separated and the aqueous phase was further extracted twice
with laO0 ml OI ethyl acetate The organic phases were combined
and waished with a saturated aqueous solution of sodium chloride
until the pH of the washings reached 4 - 5, when they were dried
:
.
,
~ - 56 -
, .
,:
'
. .: . i . ..
. . . . .. ...
, , ., - . , : , . .. .
. . . ; .
~5~3~
with anhydrous sodium sulphatc. On distilling off the solvent
from thc solution, there were obtained 22.1 g of 7~B-(D-5-t-
butoxycarbonylamino - 5 - carboxyvaleramido) - 3 - carbamoyloxymethyl -
7~Y- m ethoxy- 3 - c ephem -4 - carboxylic acid .
.
This compound was added to a phosphoric acid
buffer of pH 7. 0 containing 10 g of 5-mercapto-1-methyl-lH-
; ` tetrazole, and the mixture was agitated at 95C. for 30 minutes,:, . . .
after which the pH was adjusted to 2. 5 by addition of hydrochloric
acid, with ice-cooling. The reaction mixture was then extracted
with ethyl acetate, and the extracts were washed with a saturated
- aqueous solution of sodium chloride until the pH of the washings
reached 4 - 5, after ~vhich the extracts were dried over anhydrous
sodium sulphate. A solution of 20 g of diphenyldiazomethane
l in ether was added to the dried extracts and the mixture was
; ,~ 15 agitated for 2 hours, aiter which it was washed first with a 20%
, ~ :
a~ueous solution of sodium chloride and then with a S~ aqueous
solution of sodium bicarbonate. The washed mixture was dried
; ~ over anhydrous sodium sulphate and the solvents were distilled
off; the residue was then dissolved in chloroform, adsorbed on
a column packed with silica gel and eluted with chloroform
~` containing 1% y/v of methanol. Distil~ing off the solvent from
,
the eluent gave dLben~hydryl 7~B-(D-5-t-butoxycarbonylamino-5-
.
. . .
~5~i3:3L
:` .
carboxy~aleramido)-7~-methox~y-3-(1-methyl-l:EI-tetrazol-5-yl)
thiomethyl - 3 - c ephem -4 - carboxylate .
5 mmole of this compound were dissolved in 50 ml
of chloroform, and the solution was added to a mixture of 2. 26 g
of chloroacetyl chloride and 2. 26 g OI bis-(trimethylsilyl)trifluoro-
~ acetamide, which had previously been allowed to stand at room
-' temperature for 30 minutes, and the resulting mixture was allowed
to stand at 40C for 100 hours. The reaction mixture was then poured
into a 5% aqueous solution of sodium bicarbonate and the mixture
; 10 was agitated for 30 minutes. The organic phase was collected
and washed with a 20% aqueous solution of sodium chlox ide,
"
~, and then dried over anhydrouæ sodium sulphate, The solvent
was distilled off ~rom the vvashed solution and the residue was
dissolved in 5 ml of anisole and 10 ml of trifluoroacetic acid.
I, 15 This solution was shaken at room temperature for 5 minutes
and then evaporated to dryness under reduced pressure. The
residue was dissolved in lM phosphoric acid buffer (pH 7. 5) and
extracted with ethyl acetate. The aqueous phase was collected
, ~ and its pH adjusted to 2. 5 by addition of lN hydrochloric acid;
: . :
it was then extracted wlth ethyl acetate. The extract was dried
; ,~ over anhydrous sodium sulphate, and the solvent was distilled
, : '
' ' . ;
' ~ '
:, , .:
" ~, . . . .
j3~
off, giving 7,B-chloroacetamido-7c~-methoxy-3-(1-methyl-lH-
tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid in the
form of an amorphous powder.
'
~,
, ,:
, :
( I .
;, :
'' :
''.
',' ~ ' . ~
.. . .
.~ ~
.~ - 59 -
