CEPHALOSPORINS

CEPHALOSPORINES

English Abstract

BERGES CASE 13-18-25
ABSTRACT
The compounds of this invention are cephalosporins
having various acyl substituents at the 7-position and a
sulfaminoalkyl substituted tetrazolylthiomethyl group at the
3-position of the cephem nucleus and intermediates for the
preparation thereof.

Claims

The embodiments of the invention in which an
exclusive property or privilege is claimed are defined as
follows:
1. A process for preparing a compound of the
formula:
<IMG>
in which:
W is hydrogen or methoxy;
R1 is an acyl group selected from the group
consisting of:
<IMG> , <IMG> and <IMG>
where:
X is thienyl, dihydrophenyl, phenyl or phenyl
monosubstituted with hydroxy, hydroxymethyl, formamido,
ureido or carboxymethylamino;
A is NH2, OH, COOH or SO3H; or formyloxy when X
is phenyl;
Y is thienyl, tetrazolyl, sydnone, cyano or
aminomethylphenyl;
Z is methyl, trifluoromethyl, trifluoroethyl,
cyanomethyl or pyridyl;
m is zero to two; and
n is two to five,
or a non-toxic pharmaceutically acceptable salt thereof,
- 28 -

comprising reacting a compound of the formula:
<IMG>
where R1 and W are defined as above or a salt thereof,
with a compound of the formula:
<IMG>
where n is defined as above or a salt thereof,
or, when W is hydrogen, acylating a compound of the formula:
<IMG>
where n is defined as above and R4 is hydrogen or a
protecting ester group,
with a compound of the formula R1-OH or an activated
derivative thereof, suitably protected as necessary, followed
by removal of the protecting group(s) when necessary,
acidifying or otherwise converting to the free acid when
necessary and optionally converting the product to a non-
toxic pharmaceutically acceptable salt.
2. A process as claimed in claim 1 in which n is two.
- 29 -

3. A process as claimed in claim 1 in which R1 is
<IMG>, X is phenyl and A is NH2 or OH.
4. A process as claimed in claim 3 in which W is
hydrogen.
5. A process as claimed in claim 3 in which W is
methoxy.
6. A process as claimed in claim 1 in which R1 is
<IMG> and Y is thienyl or tetrazolyl.
7. A process as claimed in claim 6 in which W is
hydrogen.
8. A process as claimed in claim 6 in which W is
methoxy.
9. A process as claimed in claim 1 in which R1 is
<IMG> , Z is trifluoromethyl and m is zero to two.
10. A process as claimed in claim 9 in which W is
hydrogen.
11. A process as claimed in claim 9 in which W is
methoxy.
12. A process as claimed in claim 4 for preparing
7-mandelamido-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
3-cephem-4-carboxylic acid comprising reacting 7-mandelamido-
cephalosporanic acid sodium salt with 1-(2-sulfaminoethyl)-
tetrazole-5-thiol disodium salt and then acidifying.
13. A process as claimed in claim 5 for preparing
7.beta.-mandelamido-7.alpha.-methoxy-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting
7.beta.-mandelamido-7.alpha.-methoxycephalosporanic acid, sodium
bicarbonate and 1-(2-sulfaminoethyl)tetrazole-5-thiol
disodium salt and then converting to the free acid.
- 30 -

14. A process as claimed in claim 7 for preparing
7-(2-thienylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting
7-(2-thienylacetamido)cephalosporanic acid with 1-(2-sulfamino-
ethyl)tetrazole-5-thiol disodium salt and then converting to
the free acid.
15. A process as claimed in claim 7 for preparing
7-(1-tetrazolylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting
7-(1-tetrazolylacetamido)cephalosporanic acid sodium salt
with 1-(2-sulfaminoethyl)tetrazole-5-thiol disodium salt
and then acidifying.
16. A process as claimed in claim 8 for preparing
7.alpha.-methoxy-7.beta.-(2-thienylacetamido)-3-[1-(2-sulfaminoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising
reacting 7.alpha.-methoxy-7.beta.-(2-thienylacetamido)cephalosporanic
acid sodium salt with 1-(2-sulfaminoethyl)tetrazole-5-thiol
disodium salt and then converting to the free acid.
17. A process as claimed in claim 8 for preparing
7.alpha.-methoxy-7.beta.-(1-tetrazolylacetamido)-3-[1-(2-sulfaminoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising
reacting 7.alpha.-methoxy-7.beta.-(1-tetrazolylacetamido)cephalosporanic
acid, sodium bicarbonate and 1-(2-sulfaminoethyl)tetrazole-5-
thiol disodium salt and then converting to the free acid.
18. A process as claimed in claim 10 for preparing
7-trifluoromethylthioacetamido-3-[1-(2-sulfaminoethyl)tetrazol
5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting
7-trifluoromethylthioacetamidocephalosporanic acid sodium salt
with 1-(2-sulfaminoethyl)tetrazole-5-thiol disodium salt and
then acidifying.
- 31 -

19. A process as claimed in claim 11 for preparing
7.alpha.-methoxy-7.beta.-trifluoromethylthioacetamido-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
comprising reacting 7.alpha.-methoxy-7.beta.-trifluoromethylthioacetamido-
cephalosporanic acid, sodium bicarbonate and 1-(2-sulfamino-
ethyl)tetrazole-5-thiol disodium salt and then converting to
the free acid.
20. A compound of the formula:
<IMG>
in which:
W is hydrogen or methoxy;
R1 is an acyl group selected from the group
consisting of:
<IMG> , <IMG> and <IMG>
where:
X is thienyl, dihydrophenyl, phenyl or phenyl
monosubstituted with hydroxy, hydroxymethyl, formamido,
ureido or carboxymethylamino;
A is NH2, OH, COOH or SO3H; or formyloxy when X is
phenyl;
Y is thienyl, tetrazolyl, sydnone, cyano or
aminomethylphenyl;
Z is methyl, trifluoromethyl, trifluoroethyl,
cyanomethyl or pyridyl;
- 32 -

m is zero to two; and
n is two to five,
or a non-toxic pharmaceutically acceptable salt thereof, when
prepared by the process of claim 1 or its obvious chemical
equivalent.
21. A compound of claim 20 in which n is two when
prepared by the process of claim 2 or its obvious chemical
equivalent.
22. A compound of claim 20 in which R1 is
<IMG> , X is phenyl and A is NH2 or OH when prepared by the
process of claim 3 or its obvious chemical equivalent.
23. A compound of claim 20 in which R1 is
<IMG> , x is phenyl, A is NH2 or OH and W is hydrogen when
prepared by the process of claim 4 or its obvious chemical
equivalent.
24. A compound of claim 20 in which R1 is
<IMG>, X is phenyl, A is NH2 or OH and W is methoxy when
prepared by the process of claim 5 or its obvious chemical
equivalent.
25. A compound of claim 20 in which R1 is
<IMG> and Y is thienyl or tetrazolyl when prepared by the
process of claim 6 or its obvious chemical equivalent.
33

26. A compound of claim 20 in which R1 is
<IMG> and Y is thienyl or tetrazolyl and W is hydrogen
when prepared by the process of claim 7 or its obvious chemical
equivalent.
27. A compound of claim 20 in which R1 is
Y-CH2-C- and Y is thienyl or tetrazolyl and W is methoxy when
prepared by the process of claim 8 or it obvious chemical
equivalent.
28. A compound of claim 20 in which R1 is
<IMG> , Z is trifluoromethyl and m is zero to two when
prepared by the process of claim 9 or its obvious chemical
equivalent.
29. A compound of claim 20 in which R1 is
<IMG> , Z is trifluoromethyl, m is 0 to 2 and W is
hydrogen when prepared by the process of claim 10 or its obvious
chemical equivalent.
30. A compound of claim 20 in which R1 is
<IMG> , Z is trifluoromethyl, m is 0 to 2 and W is
methoxy when prepared by the process of claim 11 or its obvious
chemical equivalent.
31. The compound 7-mandelamido-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid when
prepared by the process of claim 12 or its obvious chemical
equivalent.
32. The compound 7.beta.-mandelamido-7.alpha.-methoxy-3-
[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-
carboxylic acid when prepared by the process of claim 13 or
its obvious chemical equivalent.
34

33. The compound 7-(2-thienylacetamido)-3-[1-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic
acid when prepared by the process of claim 14 or its obvious
chemical equivalent.
34. The compound 7-(1-tetrazolylacetamido)-3-[1-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic
acid when prepared by the process of claim 15 or its obvious
chemical equivalent.
35. The compound 7.alpha.-methoxy-7.beta.-(2-thienylacetamido)-
3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-
carboxylic acid when prepared by the process of claim 16 or
its obvious chemical equivalent.
36. The compound 7.alpha.-methoxy-7.beta.-(1-tetrazolyl-
acetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-
cephem-4-carboxylic acid when prepared by the process of
claim 17 or its obvious chemical equivalent.
37. The compound 7-trifluoromethylthioacetamido-3-
[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-
carboxylic acid when prepared by the process of claim 18 or
its obvious chemical equivalent.
38. The compound 7.alpha.-methoxy-7.beta.-trifluoromethyl-
thioacetamido-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
3-cephem-4-carboxylic acid when prepared by the process of
claim 19 or its obvious chemical equivalent.

Description

1~7~38
1 This invention relates to a new series of cephalosporin
compounts which have antibacterial acti~ity when administered
parenterally and to intermediaees for the preparation thereof. In
particular, the structures of the biologically active cephalosporin
compounds of this invention are characterized by having a
sulfaminoaLkyl substituted tetrazolylthiomethyl group at the
3-position of the cephe~ nucleus. Also, this invention extends
to methots and compositions for treating certain bacterial infections
using these new compounts as well a~ to certain chemical inter~ediates
and methot~ for preparing the compounds described here~ter.
The compounts of this invention are represented by the
follo~ing structural for3Lla:
" ' 1 ~ _~S .,
o ~ca2S ~ ¦ ¦
: N_ N
; cooa I ~
(C~2)n~N~S03a t
FOR~ULA I
in which:
Rl is an acyl group selected from the group consisting
. of:
g-ca-c- , Y-C~2-C- and Z-S(O)m-Ca2-C-
where:
X is thienyl, tihydrophenyl, phenyl or phenyl
monosubstituted wich hydroxy, hydroxy~ethyl, for~amido,
ureido or carboxymethylamino;
A is Na2, oa, cooa or S03-d; or rormyloxy when X is phenyl;
30 Y is thienyl, tetrazolyl, sydnone, cyano or aminomethylpnenyl;
.
. ~ .1
~l

~071188
1 Z is methyl, trifluoromethyl, trifluoroethyl, cyanomethyl
or pyrldyl;
m i9 zero to two; ant
n 19 two to five,
or a non-toxic pharmaceutically acceptable salt thereof.
It wlll be recognized that the 4-carboxylic acit group
of the compounts of Formula I may be reatily esterifiet by methods
well known to the art. These esters include, for example, simple
alkyl ant aryl esters as well as esters which are easily cleaved,
within the boty, to the parent acit such as intanyl, pivaloyloxymethyl,
acetoxymethyl, propionyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl
and thienylglycyloxymethyl esters and others. Of course, when A is
COO~, this group may be similarly esterified. All ~uch esters are
included within the scope of this invention.
A selected group of compounts of this invention is represented
by Formula I where n is two.
Another group of compounds of this invention is represented by
Formula I where n ls two, ~ ls phenyl, A is ~2 or OH, Y is thienyl or
tetrazolyl, Z is trifluoromethyl and m is zero.
E~amples of representative 7-acyl substituents (RlNH) of the compounds
of Formula I are listed below:
-hydroxyphenylacetamito
a-aminophenylacetamito
~-amino-4-hytroxyphenylacetamito
2S trifluoromethylthioacetamito
methylthioacetamido
-carboxythienylacetamito
a-carboxyphenylacetamito
~-sulfophenylacetamido
a-amino-4-carboxymethylaminophenyl-
acetamido
,

~ 071~L88
} 2-aminomethylphenylacetamido
3-sytnoneacetamido
tetrazolylacetamido
thienylacetamido
2,2,2-trifluoroethylsulfinylacetamido
cyanoacetamldo
methylsulfonylacetamido
cyanomethylthioacetamido
4-pyrldylthioacetamido
2-pyridoneaceta~ido
4-pyridoneacetamido.
Some examples of the compounds of ~or~ula I are 7-D-mantela~ido-
3-~1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-ce hem-4-carboxylic
acld, 7-(2-thlenylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acidt 7-(1-tetrazolylacatamido)-
3-~1-(2-sulfaminoethyL)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic
acid and 7-trlfluoromethylthioacetamido-3-~1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxyllc acid.
Cephalosporin derivatives having 7-acyl substituents
as defined above are well documented in the prior art. Although
substltutlon by variously qubstituted S-heterocyclicthiomethyl
groups t-C~2S~et) at the 3-position of the cephem nucleus is also
known, no compounts containing the 3-(sulfaminoalkyl substituted
tetrazolyl)thiomethyl moiety discloset herein are believed to be
known to the art.
The compounds of Formula I are prepared by acylating
7-aminocephalosporanic acid with an appropriately protected
acylatlng agent and then displacing the 3-acetoxy group with the
deslred sulfaminoalkyltetrazole thiol or its corresponding salt
with subsequent removal of the protective group(s) when pre~ent.
The carboxylic acid group of the acylating agent is activated by

07~88
any of the standard methods such as conversion to the mixed anyhdrlde,
acid chloride, acid imidazolide or activated ester. In addition,
a reagent such as dlcyclohexylcarbodiimide can be used provided
that the carboxyl group on the cephem nucleus is protected with
S an easily removable protecting group such as a benzhydryl, e-butyl,
trichloroethyl, benzyl, benzyloxymethyl, p-methoxybenzyl or
p-nitrobenzyl eRter. ~hen A is NH2, the a-amino group of the
acylating agent is, preferably, protected prior to acylation with
an easily removable protective group known in the art such as
t-butoxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl,
the methyl acetoacetate adduct or similar groups co~monly used
i~ the ~ynthesis of peptides.
The sulfaminoalkyltetrazole thlols of the formula:
N - N
LS
(CH2)~-N~S0
FORMULA II
in which n is two to flve,
are also ob~ects of this lnvention, being important intermediates
for producing pharmaceutical end products as described herein.
Alternatively, the compoundY of Formula I are prepared by
acylation of an approprlate 7-amino-3-~ulfaminoalkyltetra201ylthiomethyl
cephalosporin nucleus of For~ula III:
25~ ll
CoOR4 - N
(CH2)n-NHS03H
FORMULA III

~071~88
in which:
n is two to five; and
- R4 is hydrogen or a protecting ester group,
with an approprlate acylating agent followed by removal of the
S protective groups when present.
The compounds of Formula III above are also considered as
ob~ects of this invention.
The protective groups can be removed according to
! methods well known eo the art, such as with trifluoroacetic acid
when t-butyl or t-butoxycarbonyl protective groups are used.
The resulting salt is converted to the zwitterionic product or
to the free acid by means of an ion exchange resin such as
polystyrene-amine ion exchange resin (Amberllte IR-45) or else
by basification of an aqueous solution of the salt.
15The acylatin~ agents used as starting materials are
either know~ or prepared by known methods.
The 7-a~ino-3-sulfaminoalkyltetrazolylthiomethyl cephalosporin
starting materials of Formula III are prepared from reaction of
7-formamidocephalosporanic acid, prepared by reactio~ of 7-amino-
cephalosporanlc acid with formic acid and acetlc anhydrlde, and a20
substltuted tetrazole thiol of Formula II followed by treatment with
acld such as hydrochloric acld to remove ehe formyl group.
The sulfaminoalkyltetrazole thiols of Formula II are
prepared by reaction of the corresponding 1-aminoalkyl-5-(2,4-
dinitrophenylthio~tetrazole compounds, prepared from 2,4-dinitro-
"fluorobenzene and a l-acetamidoalkyltetrazole-;-thiol followed
by acid hydrolysis of the acetamido moiety, with sulfur
trioxide-trimethylamine complex with subsequent cleavage of the
2,4-dinitropbenyl protecting group. The l-acetamidoalkyltetrazole-
5-thiols are prepared by reaction of an acetamidoalkyldithiocarbamate
such as methyl 2-acetamidoethyldithiocarbamate with an azide such
- 6 -

1071~88
1 as sotium azide. The aceta~ldoal~yldithiocarbamates are prepared
by treatment of a N-aminoalkylacetamide such as N-(2-aminoethyl)-
acetamite with carbon disulfide and an alkyl halide such as methyl
iodide ln the presence of a base such as triethylamine.
Certain compounds of this invention are capable of
forming sal~s with, for e~ample, the alkali metals such as
sodium or potassium, the alkaline earth metals such as calcium
or with the ammonium cation. When A of Formula I is ~2~ the
compounds can exist as the zwitterion or as either an acid or
base salt. These salts are prepared by standard methods using
a wide variety of non-toxic pharmaceutically acceptable acids
and bases known iQ the art. Salts of the compounds of Formulas
I, II and III are constdered as ob~ects of this invention.
It will be recognized that due to the asymmetric
lS ~-carbon atom in the 7-acetamido group of Formula I when Rl i9
x-ca-c-, optical isomers will e~ist. Racemic or resolved products
are obtained depending upon wheeher a racemic or resolved sidechain
acid i9 used as an acylating agent. The resolved sidechain acids
are readily obtained from the racemic compounds by resolution
according to well known methods, including fractional crystallization
of a salt formed with an optically active acid or base. All of
the isomers, including separated isomers and mi~tures thereof,
are included ~ithin the scope of this invention.
The compounds of Formula I have antibacterial activity
2S against both Gram-positive and Gram-negative organisms. ~inimum
inhibitory concentrations (MIC's) range from 0.2 to >200 ~g/ml in
in vitro testing. Test results for representative compounds
are given in Table 1 below. In vivo mouse protection data
(ED50's) is given in Table 2. ~ames corresponding to compound
numbers are given in Table 3.

1071~88
TABLE 1
~IC (~g/ml) in vitro
i
Com~ound Number
Bacterla _ 1 _ 23 _ _ _ 4
S. aureuQ E~ 127 3.1, 3.1 1.6 3.1 1.6
S. aureus SR 23390 0.8, 0.8 0.4 3.1 1.6
S. villaluz Sg 70390 50, 200 25 > 200 > 200
Serep. faecali3 E~ 34358 100, 50 12.5 50 100
o E. coli SX 12140 0.8, 0.8 3.1 0.8 0.8
E. col~ E~ 33779 3.1, 1.6 12.5 0.8 1.6
Rleb. pneumo. SX 4200 1.6, 0.8 3.1 0.8 0.8
Kleb. pneumo. SX 1200 0.4, 0.2 0.8 0.4 u.2
Sal~onella ATCC 12176 0.2, 1.6 12.S 0.4 0.4
Shigella-H~ 117 0.4, 0.2 ~ 0.4 0.8
Pseudo. aerug. E~ 63 > 200, ~ 200> 200> 200 > 200
Serratia marc. ATCC 13880 100, 100> 200 200 lûO
; Protea~ morgani 1791.6, 3.1 > 200 200 > 200
Entero. aerog. ATCC 13048 50, 6.3 25 3.1 6.3
Entero. cloacae EH 31254 1.6, 1.66.3 0.8 1.6
.
-- 8 --

1071~88
TABLE 2
EDso (m~lkg) in vivo
E. coli SR 12140 Kleb. pneumo. S~ 4200
Compount ~umber 9. C-p . O. ~ . C. p . O .
1 0.46 50 0.46 ~
2 1.02 > 50 - --
3 1.56 -- --
0 4 1.82 - 50
TABLE 3
Compound Number Compount Name
1 7-D-mantel~m~do-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl~-3-caphem-4-carboxylic acit
2 7-(2-thienylacetamito)-3-~1-(2-sulfa~inoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic
acit
3 7-Cl-tetrazolYlacetamito)-3-[l-(2-sulfamizoethyl)
tetrazol-5-ylthiomethyl]-3-cephem-4-car~oxylic
acid
4 7-erifluoromethylchioacetamito-3-[1-(2-~ulfa~ino-
ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-
carboxylic acid
-

1071~88
Pharmaceutical compositions havlng antibacterial activity
which comprise a pharmaceueical carrier containlng an active but non-toxic
quantity of a compound of Formula I as well as methots of combatting
bacterial infections by administering such a composition to an infected
host in a non-toxic amount sufficient to combat such infections are also
ob~ects of thls inYentiOn. The administration may be by parenteral
in~ectlon such as subcutaneously, intramuscularly or intravenously.
The injection of sultably prepared sterile solutions or suspensions
containing an effective, non-toxic amount of the new cephalosporin
compound is the preferred route of administration.
The compounds of Formula I are formulated and administered
in the same manner as other cephalosporins. The dosage regimen
comprises adminlstration, preferably by injection, of an active but
non-toxic quantity of a compound of Formula I selected from the dosage
: L5 unit range of from 100 to 1000 mg ~ith the total daily dosage regimen
being from 400 mg to 6 g. The precise dosages are dependent upon the
age and weight of the snb~ect and on the infection being treated and
can be determined by those skilled in the art based on the data
disclosed herein compared wlth that available to the art attained with
know~ cephalosporins.
Also considered within the scope of this invention are the
7a-methoxy analogs of the compounds of Formula I, which compounds are
represented by the following structural formula:
OC~
25 ~ ~ ~ N
O N - N
COOH
(CE2) -NHS03H
FORMULA IV
or a non-toxic pharmaceutically acceptable salt thereof, in which
and n are as previously defined hereabove.
-- 10 --

10~ 38
A selected group of the compounds of Formula IV are those
where n is two.
; Representatlve of the compounds of Formula IV are 7a-methoxy-
7B-(2-thlenylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
3-cephem-4-carboxylic acid, 7a-methoxy-7~-trifluoromethylthioacetamido-3-
[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carboxylic acid,
7B-D-mandelamido-7a-methoxy-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
3-cephem-4-carboxylic acid and 7-methaxy-7,~-(1-tetrazolylaceta~ido)-3-
~1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid.
The compounds of Formula IV are preferably prepared by displacing
the 3-acetoxy group from a 7a-metho~y-7~-acylaminocephalosporanic acid or
qalt thereof, suitably protected as necessary, with a substituted tetrazole
thiol of Formula II, or a corresp~nding salt, ~ith subsequent removal of
the protective group(s) and conversion of any salts to the corresponding
free acids, all as descrlbed hereinabove. The 7~-methoxy-7~-acyla~ino-
cephalosporanic acids or salts are either known to the art or are
prepared by known methods.
As with the compounds of Formula I, all non-toxic pharmaceutically
acceptable salts and ail isomers, including separated isomers and mixtures
thereof, of the compounds represented by Formula IV are included within
ehe scope of this inveneion.
The compounds of Formula IV have anti-bacterial activity against
both Gra~-positive and Gram-negative organisms. They are ad~inistered and
formulated in the same manner as previously described for the compounds of
Formula I.
The following examples illustrate the invention but are not
to be construed as limiting the scope thereof. Temperatures are in
degrees Centigrade (~C.) unless otherwise stated.
EXA~PLE 1
7-D-~andelamido-3-[1-(2-sulfaminoethyl)tetrazol-;-ylthiomethyl]-3-cephem-
4-carboxYlic acid

107~88
To a solution of 20.4 g (0.20 mol) of N-(2-amlnoethyl)aceeamide
in 200 ml of 95~ ethanol was added 27.9 ml (0.20 mol) of triethylamine
and 12.0 ml (0.20 mol) of carbon disulfide. The exothermic reaction
reached reflux and then cooled to ambient temperature over a 1.5 hour
period. Methyl iodide (28.4 g, 0.20 mol) was added which again produced
an exothermic reaction. After 1.75 hours the reaction mi~ture was
evaporated to dryness and the solid residue was dissolved in 200 ml of
water. The aqueous solution was extracted twice with 250 ml portions
of ethyl acetate. The extracts were combined, shaken with sodium
thio3ulfate, dried (MgSO4) and evaporated to dryness to give methyl
2-acetamitoethyldithiocarbamate.
To a solution of 38.4 g (0.198 mol) of methyl 2-aceta~ido-
ethyldithiocarba~ate in 100 ml of 95% ethanol was added a solution of
13.5 8 (0.208 mol) of sodium azide in 100 ml of water. The reaction
mixture was refluxed for 24 hours then cooled and concentrated under
reduced pressure to about half volume. The solutlon was cooled to 15
and 50 ml of 6N sulfuric acid was added. The acidic solution was
filtered and the filtrate was concentrated to about 100 ml and chilled
at 5 to induce crystallization of 1-(2-aceta~ldoethyl)tetrazole-5-
thiol which wa~ collected by filtration, mp 139-139.5~. Additional
amounts of the product were obtained by continuous extraction of the
filtrate with ethyl acetate.
A solution of 9.3 8 ~0.050 mol2 of 2,4-dinitrofluorobenzene
in 50 ml of acetone was added to a solution of 9.35 g (0.050 mol) of
1-(2-acetamidoethyl)eetrazole-5-thiol and 6.85 ~1 C0.050 mol2 of
triethylamine in 100 ml of acetone and ehe reaction mixture was stirred
for 1 hour. The solid material was colleceed by filtration and
recrystallized from acetonitrile eo give 1-(2-aceeamidoethyl)-5-(2,4-
dinitrophenylthio)tetrazole, mp 197-198.
A mixture of 6.5 g (0.02 mol) of 1-~2-aceta~idoethyl)-5-
(2,4-dinitrophenylthio)tetrazole, 100 ~1 of 12 N hydrochloric acid and
- 12 -

10711B8
100 ml of 95% ethanol was refluxed for 4.5 hours. The mixture was
evaporated eo dryness to give a gummy residue uhich crystallized upon
atditio~ of ethanoL to give 1-(2-amtnoethyl)-5-(2J4-tinitrophenylthio)
tetrazole hytrochloride, mp 217-219 (d).
To a solution of 3.5 g (0.01 mol) of 1-(2-aminoethyl)-5-(2,4-
dinitropherlylthio)tetrazole hydrochloride in 30 ~1 of dry ti~ethylformamide
was added 1.4 g (0.01 mol) of sulfur trioxide-tri~ethylamine cple~
followed by 1.4 ml (0.01 mol) of triethylAm~ne. ~he ~isture was
stirred for C.5 hour and then filtered. ~he filtrate was evaporated
in vacuo~ acetoue was adted to the residue, ~he precipieate was
removed by filt~ation and the ~iltrate was evaporated to dryness.
~ethanol was added to the residue and the solid material produced upon
scratching ~as re~oved by filtration. The methanolic filtrate was
broughe to pa 11.3 by addition of 5% methanolic sodium me~ho~ide, stirret
lS for 1.25 hour~, filtered ant tlluted uith 300 ml of ~ther. The resulting
solid was removet by filtraeion ant the filtrate was evaporated to
dryuess to give a resitue which uas eriturated with 95% ethanol to intuce
crystallization. The solid product was collected by filtration and
tissolved in metha~ol and the methanolic solution was concentrate~ to
10 ml, dilutet uith 75 ml of 95% ethanol and re-concentrated to S ml to
give l-(2-sulfaminoe~hyl)tetrazole-5-ehiol tisotium salt, mp 122-127~.
C3a5N503S2 2 Na 1.5 ~z
Calculated: 12.16% C; 2.72% H; ~3.64X N
Found: 12.252 C; 7. 98~ H; Z.3.77~. ~
A solution of 1-(2-sulfami~oethyl)tetrazole-;-thiol disotium
~alt in water ls passet through an Amberlite ~B-L20X ion e~change
resi~ column to give, after lyophilization, 1-(2-sulfaminoethyl)-
tetrazole-5-thiol.
To a mixture of 2.11 g (0.006 mol) of 7-D-mantelamidocephalo-
sporanic acit sotium salt and 1.18 g (0.004 mol) of 1-(2-sulfaminoethyl)-
tetrazole-S-thiol disodium salt i~ 30 ml or uater was added 10~ aqueous -~
- 13 -
.".~
- ' '
,

1071~88
sodium hydroxide solution a~t then 5~ aqueous sodium bicarbonate solution
to p~ 7.3. The reaction mixture was heated at 70 ror 2.66 hours, the~
it was coolet, covered with ethyl acetate, acidified to p~ 2.5 with 3N
hytrochloric acid and e~tracted twice wieh eehyl acetate. The aqueous
phase was neutralizet to pa 7.0 by adtition of 10% aqueous sodium
hydroxide and the~ 5% aquaous sodium bicarbonate solution-~ and
chromatographed on gAD-7 resin ~ith water and methanol a~ eluants.
A~ter removing t~e methanol in vacuo the chr = tography fractions were
lyophilized to give 7-D-mandelamido-3-~1-(2-sulfaminoethyl)tetrazol-5-
ylth~omethyl]-3-cephem-4-carboxylic acid disodium salt.
Clg~lg~7ogs3-2 Na 3 ~2
Calculated: 34.08% C; 3.76~ ~; 14.64~ N
Found: 34.56% C; 3.25% ~; L3.96~ N
Ac aqueous solution of 7-D-mantelamido-3-~1-(2-sulfaminoethyl)-
tetrazol-5-ylthicmeehyl]-3-cephem-4-carboxylic acid disodium salt is
pas~ed through a column of Ambert~te 1~-120~ ion eYchange re~in co give
the title compount.
E~A~PLE 2
7-(2-Thienylacetamido)-3-~1-(2-sulfaminoethyl~tetrazole-5-ylthiomeehyl]-
3-cePhe~-4-carboxYlic acid
A mixture of 1.89 g (0.064 mol) or 1-(2-su14aminoethyl)t2trazole-
5-thiol tisodium salt and 2.67 8 (0.064 mol) of 7-(2-ehienyla~etamido)-
cephalosporanic acit sodium salt in 40 ml of water was heated at 69 for
5.5 hours while ~aintaining the pH at 7.4 by addition of dilute aqueous
sodiu~ bicarbonate solution. After cooling, ehe mi~ture was e~tracted
~S wlth ethyl acetate. The aqueous phase was neutralized, evaporated co
dryness and the residue was passed through a .YAD-4 column eluting with
water and methanol. The methanol was removed by evaporation and the
aqueous resitue was lyophilizet to give a solid material. The solid was
quspented in methanol, the insoluble material was remo~Jed by fileration
and ehe filtrate was evaporated to tryness to ~ive 7-(2-thienylaceta ido)-
3-[1-(2-sulfaminoethyl)t~trazol-5-ylthiomethyl~-3-cephem-4-carboxylic acid
` ! - 14 -

~07~88
disodium salt.
C17H17N707S4 2 Na 1 CH40
Calculated: 33.90% C; 3.31Z H; 15.37% N
Found: 34.04~ C; 3.57% ~; 14.74% N
7-(2-Thienylacetamido)-3-[1-(2-sulfamlnoethyl~tetrazol-5-yl-
thiomethyl~-3-cephem-4-carboxyllc acid disodium salt is converted to the
title compount as described in EYample 1.
E~AMPLE 3
7-(1-Tetrazolylaceta~ido)-3-~1-(2-sulfaminoethyl)tetrazole-5-yl-
thlomethvl]-3-cePhem-4-carboxYlic acid
1 A mi~ture of 3.5 g (8.5 mmol) of 7-(1-tetrazolylacetamido)-
cephalospor3nic acid sodium salt and 2.96 g (10 mmol) of 1-(2-sulfamino-
ethyl)tetrazole-5-thiol disodium salt in 50 ml of water was stirred at
65~ for 6.5 hours while maintainin~ the p~ of the reaction mi~ture at
7.0 by addition of 5% aqueous sotium bicarbonate solution. The
mixture was cooled to ambient temperature, acidified to pH 1.5 with
3N hydrochloric acid, filtered and extracted three times with ethyl
acetate. The p~ of the aqueous phase was then ad~usted to 7.0 by
adtition of sodium bicarbonate, the solution ~a~ chramatographed on
a 2AD-2 column and the resulting product was freeze-dried to give
7-(1-tetrazolylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
3-cephem-4-carboxyLic acit disodium salt.
C14~15N117S3 2 Na 2 a2o
Calculated: 26.80% C; 3.37~ ~; 24.55% N
Fount: 27.11% C; 3.40% ~; 24.18% N
7-(1-Tetrazolylacetamido)-3-[1-(2-sulfaminoethyl)tatrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acid disodium salt is converted to
the title compound as described in Example 1.
EXAMæLE 4
7-Trifluoromethylthioacetamido-3-~1-(2-sulfaminoethyl)tetrazol-5-
ylthiometh~l]-3-ce~hem-4-carboxYlic acid
A solution of 3.05 g (0.01 mol) of 1-(2-sulfaminoethyl)tetra-
- 15 -

i~7~88
zole-5-thiol disodium salt ant 4.36 g (0.01 mol) of 7-trifluoromethyl-
ehioacetamidocephalosporanic acid sodium salt in 50 ~1 of water was
heated at 70 for 5.5 hours while maintaining the pH at 7.5 with 5%
aqueous sodium bicarbonate. The reaction mi~ture was diluted with SO
ml of water and extracted twice wlth ethyl acetate. The aqueous phase
was acidified to pH 2 and extracted three times with ethyl acetate.
The aqueous layer was brought to pH 7.4 by addition of 5~ aqueous
sodium bicarbonate and the solution was passed through a XAD-4 resin
column while eluting with water followed by methanol. The methanol
solution was evaporated to dryness and the residue was dissolved in
75 ml of water. The aqueous solution was ~Ytracted twice with ether
and petroleum ether then lyophilized. The lyophilized material was
dissolved in methanol, the solvent was evaporated to dryness and
triturated with ether to give 7-trifluoromethylthioacetamido-3-rl-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carboxylic acid
disodium salt.
C14H14F3N707S4-2 Na 2 H20
Calculated: 25.49~ C; 2.75% a; 14.86%
Found: 25.85Z C; 2.78% H; 14.13% ~
7-Trifluoromethylthioacetamido-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl~-3-cephem-4-carboxylic acid disodium salt is converted
to the title compount as describet in Example 1.
E ~P~E S
7-(D-a-Aminophenylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
vlthiomethyl]-3-cePhem-4-carbo~Ylic acid
A solution of 7.58 g (0.015 mol) of 7-(,D-a-t-butoxycarbonyl-
aminophenylacetamido)cephalosporanic acid, 2.96 g (0.01 mol) of 1-(2-
sulfaminoethyl~tetrazole-5-thiol disodium salt and 1.26 g (O.OlS mol)
of sodium bicarbonate in 125 ml of water is stirred at 60 for 5 hours
~hile ~aintaining the pH at 7.0-7.2 by addition of sodium bicarbonate.
The mixture is cooled and e~tracted with ethyl acetate. The aqueous
phasa is acidified to pH 2.5 with 3N hydrochloric acid and the acidic
- 16 -

07~L~88
solution i~ extracted again with ethyl acetate. The aqueous phase
is brought to pH 7.1 by addltion of 5~ sodlum carbonate solution, then
passed through a XAD-4 ion exchange resin column and eluted-~ith water
and meehanol to give 7-(D-a-t-butosycarbonyla~inophenylacetamido)-3-
[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carbo~ylic
acid disodium salt.
7-tD-a-t-ButYoxycarbonylaminophenylacetamido)-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carbo~ylic acid disodium ~alt
i9 stirred at 25 with 25 ml of trifluoroacetic acid and 25 ml of 1,3-
dimethoxybenzene for 2.25 ho~lrs. The misture is evaporated to dryness,
ether is added to the residue and the precipitate is collected, washed
with ether, stirred in acetonitrile for 2 hours and tried in vacuo to
glve the title compound as the trifluoroacetic acid salt.
An aqueous solution of the trifluoroacetic acid 3alt is
lS brought to p~ 5.0 by atdition of dilute aqueous sotium hydroside.
After lyophilization, the lyophilized material is dissolved in meth-
anoi and ether is added to precipitate 7-(D-a-aminophenylacetamido)-
3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carbosylic
acid sodium salt. The sodium salt is dissolved in water and the aqueous
solution is passed through an Amberlite IR-1208 ion exchange resin
column. Lyophilization of the eluted material gives the title c~mpound.
E~ANPLE 6
Reaction of the N-t-buto~ycarbonyl derivative of the
following cephalosporanic acids:
7-(-amino-4-hydro~yphenylaceta~ido~cephalosporanic acid
7-(-amino-4-formamidophenylacetamido)cephalosporanic acid
7-(a-amino-3-form~midophenylacetamido)cephalosporanic acid
7-(-amino-4-ureidophenylacetamido)cephalosporanic acid
7-(~-amino-3-ureidoph&nylacetamido)cephalosporanic acid
7-(-a~ino-4-hydroxymethlyphenylacetamido)cephalosporanic
acid
^ 17 -

1071188
L 7-(a-amino-1,4-cyclohexadienylacetamido~cephalosporanic acid
7-(-amino-4-carboxymethylaminophenylacetamido)cephalo-
sporanic acid
with 1-~2-sulfaminoethyl)tetrazole-5-thiol disodium salt as described
in the procedure of Example 5 followed by removal of the protective
group and conversion of the trifluoroacetic acid salts to the free
acids as d2scribed therein gives the following compounds of this
.! invention:
7-(a-amino-4-hydroxyphenylacetamido)-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-4-formamidophenylacetamido)-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-3-formamidophenylacetamido)-3-[1-(2-sulfamino-
ethyl)tetrazol-5-ylthlomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-4-ureidophenylacetamido)-3-[1-(2-sulfamino-
ethyl)tetsazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-3-ureidophenylacetamido-3-[1-(2-sulfamino-
. ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-4-hydroxymethylphenylacetamido)-3-[l-(2-
sulfaminoethyl)tetrazol-5-ylthlomethyl]-3-cephem-4-carboxylic acid
- 7-(a-amino-1,4-cyclohexadienylacetamido)-3-[1-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(a-amino-4-carboxymethylaminophenylacetamido)-3-[1-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid.
E~AMPLE 7
~5
7-(4-aydroxymandelamido)-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acit is prepared by reaction
of 7-(4-hydroxymandelamido)cephalosporanic acid sodium salt and l-
(2-sulfaminoethyl)tetrazole-5-thiol disodium .~alt followed by treatment
of the product with Amberlite IR-120H ion exchange resin as described
3~
in the procedure of Example 1.
- 18 -

1071~88
. 1 EgAMPLE 8
- When the sodium salt of a cephalosporanic acit listed below:
7-(3-sytnoneacetamido)cephalosporanic acid
7-(2-aminomethylphenylacetamido)cephalosporanic acid
is reacted with 1-(2-sulfaminoethyl)tetrazole-5-thiol disodium salt ~ :
by the procedure described in Example 1 and the product is converted
to the free acid as described therein, the following compounds of
this invention are obtained, respectively:
7-(3-sydnoneacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthi~meehyl]-3-cephem-4-carboxylic acid
7-(2-aminomethylphenylacetamido)-3-[1-(2-sulfa~inoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid.
EXAMPLE 9
Reactlon of the sodium salt of a cPphalosporanic acid
llsted below:
7-(2,2,2-trifluoroechylthioacetamido)cephalosporanic acid
7-methylthioacetamidocephalosporanic acid
with 1-(2-sulfaminoethyl)tetrazole-5-chiol as described in the proceduse
. of Example 4 glves, after conversion of the salts formed to the
free acids, the following compounds of this invention as final
products:
7-(2,2,2-trifluoroethylthioacetamido)-3-[-(2-sulfaminoethyl)-
tetrazol-5-ylthiomethyl~-3-cephem-4-carboxylic acid
. 7-methylthioacetamido-3-~1-(2-sulfaminoethyl)tetrazol-5- 2 ylthiome~hyl~-3-cephem-4-carboxylic acid.
E~LE 10
When an equivalent amount of an N-aminoalkylacetamide listed
below:
N-(3-aminopropyl)acetamide
N-(4-aminobutyl)acetamide
N-(5-aminopentyl)acetimide
- 19 -

~(~7~88
i8 used in the procedure of Example 1 in place of N-(2-aminoethyl)-
acetamide and the resuLting dithiocarba~ates are treated with sodium
azide to produce the corresponding l-acetamidoalkyltetrazole-5-thiols
which are converted to the l-sulfaminoalkyl derivatives, all as
describet therein, the following 1-sulfaminoalkyltetrazole-5-thiols
are obtained:
1-~3-sulfaminopropyl)tetrazole-5-thiol
1-(4-sulfaminobutyl)tetrazole-5-thiol
1-(5-~ulfaminopentyl)tetrazole-5-thiol.
0 Reaction of the disotium salt of a l-sulfaminoalkyltetrazole-5-
thiol listed above with 7-D-mandelamidocephalosporanic acid sodium salt
as described in the procedure of E~ample 1 followed by conversion of the
salts formed to the free acids, gives the following compounds of this
~ invention:
i 15 7-D-mandelamido-3-[1-(3-sulfaminopropyl)tetrazol-5-ylthio-
methyl~-3-cephem-4-carboxylic acid
7-D-mantelamito-3-[1-(4-sulfaminobutyl)tetrazol-5-ylthio-
methyl]-3-cephem-4-carboxylic acid
7-D-mandelamido-3-~1-(5-sulfa~inopentyl)tetrazol-5-ylthio-
methyl]-3-cephem-4-carboxylic acid.
Likewise, reaction of the substituted tetrazol thiols or
the correspondiug disodium salt listet above with any of ~he 7-acyl
cephalosporanic acids mentioned herein or their corresponding salts
accorting to procedures described herein gives the corresponding
compounds of this invention.
E~PLE 11
Beaction of a cephalosporanic acid listed belo~ or its
corresponding salt:
7-(-hydroxy-2-thienylacetamido)cephalosporanic acid
7-(a-carboxy-2-thienylacetamido~cephalosporanic acid
7-(a-sulfophenylacetamido)cephalosporanic acid
- 20 -

with 1-(2-sulfaminoethYl)tetrazole-5-thiol disodium salt by
procedures described hereinabove gives, after conversion of the
product to the free acid, the followlng compounds of this invention:
7-(~-hydroxy-2-thienylacetamido)-3-[1-(2-sulfaminoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(-carboxy-2-ehienylacetamido)-3-~1-(2-sulfaminoethyl)-
tetrazol-5-ylthiomeehyl]-3-cephem-4-carboxylic acid
7-(a-sulfophenylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl~-3-cephem-4-carboxylic acid.
EXAMPLE 12
7-Amino-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl~-3-cephem-
4-carboxvlic acid
To a mixture of 97 g (200 ml, 2.1 mol) of formic acid,
distilled from anhydrous copper sulfate, and 37.5 ml (0.4 mol)
of acetic anhydride was added 25.0 g (0.1 mol) of 7-aminocephalosporanic
acid. The mixture was stirred at ambient temperature for 0.5 hour,
then evaporated to dryness. The residue was dissolved in ethyl
acetate and the ethyl acetate solution was filtered a~d evaporated
to dryness to give a residue which was recrystallized from
ether-petroleum ether to give 7-for~m~docephalosporanic acid.
A mixture of 1.0 g t3.3 mmol) of 7-for~amidocephalosporanic
acid and 0.7 8 (2.6 mmol~ of 1-(2-sulfaminoethyl)tetrazole-5-thiol
disodium salt in 15 ml of ~ater i9 stirred at 65-70~ for 3 hours
while maintainin8 the p~ at 7Ø The 3ixture is cooled, acidified
:!
to pa l.o with hytrochloric acid and extracted with ethyl acetate.
The extrace i9 filtered and the filtrate is evaporated to drvness
to give a residue which is dissolved ln methanol. The methanol
solution is filtered and ether is added to precipitate tne title
compound which is collected by filtration.
EXA~PLE 13
7-(4-Pyridylthioaceta~ido)-3-l1-(2-sulfaminoethyl)tetrazol-5-
vlthiomethvll-3-cePhem-4-carboxvlic acid
- 21 -

~o7~88
(4-Pyridylthio)acetyl chloride (0.53 g, 2.8 mmol) is dropwise
added to a mixture of 1.0 g of 7-amino-3~ (2-sulfaminoethYl~tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acicl and 0.9 g (9.0 mmol) of tri-
ethyla~ine in 10 ml of dry dimethylformamide. The reaction mlxture is
stirred for 1.5 hour at -10~, then it is warmed to ambient température
ant stirred for 1 hour. This mixture is filtered and the filtrate is
diluted wlth 200 ~1 of ether-petroleum ether. The precipitate is col-
lected by filtration and dissolved in methanol. The methanol solution
ls filtered and the filtrate is evaporated to dryness to give the title
lQ compound as itq corresponding sodium salt.
7-(4-Pyridylthioacetamido)-3-[1-(2-sulfaminoethyl)-
tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid sodium salt
is converted to the title compound by procedures described above.
EXAMPLE 14
Acylation of 7-amino-3-[1-(2-sulfaminoethyl)tetrazol-5-
ylthiomethyl]-3-cephem-4-carboxylic acid with an activated
derivatlve of the following acids:
cya~oacetic acid
3-pyridylthioacetic acid
cyanomethylthioacetic acid
2,2,2-trifluoroethylsulfinylacetic acid
methylsulfonylacetic acid
2-pyridone-~-acetic acid
4-pyritoue-N-acetic acid
as describet ln the procedure of Example 13 gives the following
compounds of this invention:
7-cyanoacetamido-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(3-pyridylthioacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-cyanomethylthioacetamido-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acid
- 22 -

- ~7~88
7-(2,2,2-trifluoroethylsulfinylacetamido)-3-[l-(2-
sulfaminoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-methylsulfonylaceta~ido-3-~1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carboxylic acid
7-(2-pyridoneacetamido)-3-11-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl~-3-cephem-4-carbo~ylic acid
7-(4-pyridoneacetamido)-3-[1-(2-sulfaminoethyl)tetrazol- -
5-ylthiomethyl~-3-cephem-4-carboxylic acid.
EXAMPLE 15
! o 7-(D-a-Formyloxyphenylacetamido)-3-~1-(2-sulfaminoethyl)tetrazol-
! l 5-ylthiomethyl]-3-cephe~-4-carboxylic acid
7-~mino-3-~1-(2-sulf2minoethyl)tetrazol-5-ylthiomethyl~-
3-cephem-4-carbo~ylic acid is reacted with the formate ester of
D-mandeloyl chloride according to the proceture of Example 13 to
give the title compound.
E~AMPLE 16
7a-Methoxy-7~-(2-thienylacetamido)-3-[1-(2-sulfaminoethyl)tetrazol-5-
YlthiomethYll-3-cePhem-4-carboxylic acid
A solution of 1.28 g (3 mmol) of 7a-methoxy-7~-(2-thienylace-
tamido)cephalosporanic acid sodium salt is dissolved in 50 ml of water,
1.33 g (4.5 mmol) of 1-(2-sulfaminoethyl)tetrazole-5-thiol disodium salt
is added and the solution is heated at 70 untiL thin layer chromatography
indicates consu~ption of the starting material (ca. 5 hours). The reaction
mixture is chromatographed on XAD-4 ion exchange resin wlth, after washing
with water, methanol as eluant. Evaporation of the methanol solution gives
the title compound as t&e disodium salt.
7a-~ethoxy-7~-(2-thienylacetamido)-3-[1-(2-sulfaminoethyl)tetra-
zol-5-ylthiomethyl~-3-cephem-4-car~oxylic acid disodium salt is converted
to the title compound as described above.
EXAMPLE 17
7a-~ethoxy-7~-trifluoromethylthioacetamido-3-[1-(2-sulfaminoethyl)tetrazol-
5-YlthiomethYl]-3-cephem-4-carboxylic acid
- 23 -

~07~188
To a cold solution of 5.25 g (0.012 mol) of 7B-amino-7a-
methoxycephalosporanic acid benzhydryl ester in 200 ml of methylene
chloride containing 1.79 8 (0.012 mol) of N,N-diethylaniline is added
dropwise over a 20 minute period a solution of 1.82 g (0.012 mol) of
trifluoromethylthioacetyl chloride in 50 ml of methylene chloride.
After stirring for 30 minutes, the mixture is extracted successively
wlth 5Z aqueous sodium bicarbonate, 5~ aqueous hydrochloric acid and
finally with brine. The organic phase is dried (MgSO4) and the solvent
evaporated to give 7a-methoxy-7B-trifluoromethylthioacetamidocephalo-
sporanic acid benzhydryl ester.
7a-Methoxy-7~-trifluoromethylthioacetamidocephalosporanic
acid benzhydryl ester is dissolved in a cold mixture of trifluoroacetic
acid-anisole (2:1) and the mixture is stirred for 1.5 hour without ex-
ternal cooling. The solvent is evaporated in vacuo and the residual
product is taken up in ethyl acetate, washed with water, dried (~gSO4)
and concentrated _ vacuo to a small volu~e. This solution is added
dropwise to stirred petroleum ether to precipieate 7a-methoxy-7B-tri-
fluoromethylthioacetamidocephalosporanic acid.
7a-Methoxy-7~-crifluoromethylthioaceeamidocephalosporanic
acid (2.2 g, 5 mmol) is suspended in 75 ml of water and 0.4 g of solid
sodium bicarbonate is added until solution is complete. To this solution
is added 2.21 g (7.5 mmol) of 1-(2-sulfaminoethyl)tetrazole-5-thiol di-
sodium salt and the mixture is heated at 70 for 7 hours. The pH of the
reaction mi~ture is maintained at 7.5 by dropwise addition of 3N hydro-
chloric acid as necessary. Progress of the reaction is monitored by
thin layer chromatography and ~udged to be complete when tlc indicates
disappearance of starting material. The reaction mixture is chromato-
graphed on a column of ~AD-4 resin and the product is eluted from the
colu~n with methanol. Evaporation of the methanol solution gives 7a-
methoxy-7~-trifluoromethylthioacetamido-3-[1-(2-sulfaminoethyl)tetrazol-
3~
5-ylthiomethyl]-3-cephem-4-carboxylic acid disodium salt.

107~1~8
1 The disodium salt is convertet to the title compound by
procedures described hereinabove.
E2AMoeLE 18
7~-D-Mandelamido-7a-methoxy-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthio-
methy~-3-cephem-4-carboxYlic acid
A cold solution of 2.6 g (6 ~mol) of 73-amino-7a-methoxycephalo-
sporanic acid benzhydryl ester in 100 ml of methylene chloride containing
0.9 g (6 mmol) of N,~-dlethylaniline is treated dropwise over a 15 minute
period wlth a solution of 1.7 g (6 mmol) of D-0-dichloroacetyLmandeloyl
chloride in 25 ml of methylene chloride. The reaction mixture is allowed
to come to room temperature with stirring and then ls extracted successively
with 5% aqueous sodium bicarbonater 5~ hydrochloric acid and brine. The
organic phase i9 dried and evaporated in vacuo. The residue is dissolved
in colt trifluoroacetic acid-anisole (2:1) and the mixture is stirred at
ambient temperature for l hour. The mixture is evaporated in vacuo and
the residue is dissolved in 5~ aqueous sodium bicarbonate. The pa is
raised to 9-9.3 by addition of 5~ aqueous sodium carbonate and maintained
there for 30 minutes to complete cleavage of the dichloroacetyl group.
The solution is cooled in ice, layered with ethyl acetate and acidified
to pa 2.0 with dilute hydrochloric acid. The layers are separated and
after a second extraction of the aqueous layer with ethyl acetate the
organic phases are combined, dried and evaporated in vacuo to yield 7~-
D-mandelamido-7a-methoxycephalosporanic acid.
7~-D-Mandelamido-7a-methoxycephalosporanic acid (2.2 g, 5 mmol)
is suspented ln 75 ml of water and solid sodium bicarbonate is atded until
~5 all of the acid has dissolved. To this is added 2.21 g (7.5 ~mol) of
1-(2-sulfaminoethyl)tecrazole-5-thiol disodium salt and the mixture is
heated at 70~ for 7 hours. The pH of the reaction mixture is maintained
at 7.5 by addition of 3N hydrochloric acid. Chromatography of this
solution on XAD-4 resin while eluting with methanol gives, upon evapo-
ration of the methanol, the title compound as its disodium salt.
- 25 -

107~18~
7B-D-Mandelamido-7a-methoxy-3-[1-(2-sulfaminoethyl)tetrazol-
5-ylthiomethyl]-3-cephem-4-carbQxylic acid disodium salt is converted
to the title compount by procedures described hereinabove.
E~AMPLE 19
7-Methoxy-7B-(D-a-aminophenylacetamito)-3-[1-(2-sulfaminoethyl)tetrazol-
5-~ hiomethvll-3-cephem-4-carbo~Ylic acid _ _
To a solution of 5.3 g (0.012 mol) of 7B-amino-7a-methoxycephalo-
sporanic acit p-nitrobenzyl eqter in 200 ml of methylene chloride is
added 3.0 g (0.012 mol) of D-a-t-butoxycarbonylaminophenylacetic acid and
2.5 g (0.012 mol) of ticyclohexylcarbodiimide. The mixture is stirred
for 18 hours at ambient temperature then filtered. The filtrate is
evaporated in vacuo and the residue is dissolved in methanol-cetrahydro-
furan and hydrogenated over 5% palladium on carbon to give 73-(D-a-t-
butoxycarbonylaminophenylacetamido)-7a-methoxycephalosporanic acid.
7B-(D-a-t-Butoxycarbonylaminophenylacetamido)-7a-methoxy-
cephalosporanic acid (2.68 g, 5 mmol) is dissolved in 75 ml of water
by adding 0.4 g of solid sodium bicarbonate. 1-(2-Sulfamlnoethyl)tetra-
zole-5-ehiol disodium salt (2.21 g, 7.5 mmol) is added and the reaction
mixture is heated at 70 until thin layer chromatography indlcates that
the starting material has disappeared. The reaction mixture is chromato-
graphed on ~YAD-4 resin and eluted with methanol. Evaporation of the
methanol solution gives 7a-methoxy-7B-(D-a-t-butoxycarbonylaminophenyl-
acetamido)-3-~1-(2-sulfaminoethyl)te~razol-5-ylthiomechyl]-3-cephem-4-
carboxylic acit disodium salt.
The disodium salt is suspended in l:l trifluoroacetic acid -
anisole and stirred at ambient temperature for two hours. F~cess tri-
fluoroacetic acit is removed by evaporation, the residue is trituratet
with ether and the resulting precipitate is collected by filtration and
stirred with acetonitrile to give the title compound as its trifluoroacetic
acid salt.
An aqueous solution of the trifluoroacetic acid salt is brought
to pH 7 by addition of 5~ aqueous sodium bicarbonate then chromatographed
- 26 -

1~711~8 -:
: 1 on XAD-4 resin with methanol as elutant. ~he solid material obtained
after evaporation of the methanol is dissolved in water and the aqueous
solution ls passet through a cation exchange column (IR-120~). Lyophili-
zation of the eluted material gives the title compound.
S EXAMPLE 20
7a-Methoxy-7~-(1-tetrazolylacetamito)-3-[1-(2-sulf~m~noethyl)tetrazol-5-
Ylthiomethyl]-3-cephem-4-carboxylic acid
Substitution of an equivalent amount of l-tetrazolylacetyl
chlorite in the procedure of Example 17 ~i-es 7~-methoxy-7~-(1-tetrazolyl-
acetamldo)cephalosporanic acid benzhydryl ester which is converted to
7~-methoxy-7~-(1-tetrazolylacetamido)cephalosporanic acid as described
therein.
Reactlon of 7-methoxy-73-(l-tetra201ylacetamido)cephalosporanic
acit, 1-(2-sulfaminoethyl)tetrazole-5-thlol disodium salt ant sodium bi-
carbonate as described ln Example 17 gives, after conversion of the product
disodium salt to the free acid as described above, the title compound.
EXAMPLE 21
An injectable pharmaceutical composition is for~ed byadding sterile water or sterile saline solution (2 ml) to 500 mg of
7-D-mandelamido-3-[1-(2-sulfaminoethyl)tetrazol-5-ylthiomethyl]-
~0
- 3-cephem-4-carboxylic acid disodium salt.
Pharmaceutical co~positions of the other antibacterial
compounds tisclosed above may be formulated in a similar manner.
.
- 27 -
. .