CA1072078A - Cephalosporin compounds - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
The compounds of this invention are cephalo-sporins having various acyl substituents at the 7-position and a sulfonic acid or sulfamoyl substituted tetrazolyl thiomethyl group at the 3-position of the cephem nucleus and intermediates for the preparation thereof. The 7-acylated compounds have antibacterial activity.

Description

~Lot~Z~8 ntion compri~es a new ~eri~s of `
cephalospori~ compounds ~hich h~ve antibacterial aetivity whe~ administared paren~erally and to intermedlates for ~-the prepara~ hereof . In particular g the ~trueture~
of the blnlogically active cephalo~porin compounds o~ ~h invention are char~cterized by having a ~ulfonic acid or .;
sulfamoyl ~ubstituted tetrazolyl thi~ethyl group at the 3-position of ~he cephe~ nucl~us. Al~o, thi~ invention `~
ex~ends to m~thods ~nd compositfon~ for treatirlg certa~n bacterial infection~ using these new ca2pourld~ as well as i~.
to cer~ain ch~snical intermediate~ and methods ~r pre~
pari~g.the compound~ described here~fterO
The compound~ of this irlvention are represented ;

by the ollowing structural formula:

R3?~--r,~5 o ~ CH2 COOH
~ )n~S2R ~ `
PO~.MULA I
in which:
each individual Rl i8 hydro~en or lower alkyl;
n i8 ~ne to ~

2 S * i8 hydroxy, ~mino, lower alkylamino or di( lower~a~ylsmi~o; ~nd E3 18 ~n acyl group ~elected i~OM the group con~isting of~
O O o -~' x-cH-a~ ~Y ~2 ~ and 2~ )m~CH2~Co ' ~ ' ', : " :

~here:
X i~ ~hl~nyI; dIhydrophenyl; phenyl; phenyl - ~;
mo~o-~ubs~ituted with hydroxy, hydro~ymethyl, fosmamido j ureido or c~rboxymethylamlno; or 3 ~luoro-4-hydroxyphenyl;
A is ~2~ OH9 COO}~ or SO3~; or formyloxy when X i~ phenyl;
Y i~ thienyl, tetrazolyl J cyano, sy~none or . :~
amL~o~thylphenyl; :
., ,~:
Z i8 methylJ trifluoroma~hyl, trifluoroethyl, cyanom~t~yl or pyr idyl; ~nd ~ ;
m i~ zero to two, ;
or a non-toxic pharmaceutically acceptable salt thereof, ~
A8 u~2d her2~n, the term 'llower alkyl" refer~ :.

~o grou~s h~ving, from one ~o four ~arbon atoms, preferably -~ -methyl and ethyl.
It will be recogniz~d ~chat the 4-carboxyliic acîd group of the colapounds of For~ula I may be readily esteri fied by method~ well known to the art. The~e e~ters :
inelude, or example, ~imple alkyl and aryl esters as well as es~ers which are ea~ily cleaved, wlthin the body, to the parent acld ~uch a~ indanyl, pivaloylo~ymethyl, acetoxymethylg propionyloxyme~hyl, glycyloxymethyl, phenyl- .
glycylo~ymethyl and thienylglycyloxymethyL e~ters and ~`.
other~. Of collrse, when A ig COOH~ this group may be similarly esterifled. All ~uch e~'cers are i~cluded within the ~cope of thi~ :Lnvention. :
Prefer~ed co~pound~ of ~his inve~ion are ~ -repre~nted by Fomlula I where Rl i~ hy~rog~n; n is one ~o - ~.

3~ 2 five; R i~ hydroxy, am~no) lower slkylam~o or di(lower)~
al~ylamino; R i~ X~H- ~, Y-CH2- o or Z-5 ~O)m~CE12-~,:

-- 3 ~

~o~zo~s X 1~ thie~yl, dihydrophenyl, ph~nyl, phenyl m~no-sub~ `
st~tut~d with hydroxy, hydroxym~thyl, ~or~nido, ur~i~o or carboxy~ethylamirlo, or 3-fluoro-4-hydro~yphenyl; A
~s ~H~, OH, COOH or SO311; Y i~ thi~nyl, te~razolyl, cyanoJ
~yd~one or aminomethylphenyl; Z i8 methyl, tri~luoro-~nethyl, trifluoroe~hyl, cy~omethyl or pyridyï and m i~ zero ~co t~o.
Ad~antageous eo~po~d~ of th~s ~rlven~ion are repr~nt~d by For~l~ I w2~er~ Rl i~ hydrogen; n i8 one to five; R ~ hydro~, amîno, lower alkylamialo or di(lower)~
alkyl~mLIlo~ X i8 phen3rl or hydroxyphe~Lyl; A i8 NH2 or ~H; ~- ;
Y i~ tbie~yl; Z i8 ~ethyl, trifluor~ethyl or trifluoro-ethyl and m i8 z~ro to two.

~t ad~ra~tageous are the compotmd~ represented by Formula I where Rl ls hydrogen; n i8 one to five; R2 i~ ~ydro~ or amlno; X i8 phe~yl or 4-h~droxyphenyl;
.
A i~ NH2 or OH; Y i~ thie~yl; Z i~ trifluoromethyl and ::
i8 ~e~o.
~x~ple3 o~ the mo~t pre~srr~d 7-~cyl sub~
" , .
stituents (~NH-) of the ccmpounds of Formula I are listed belo~:
a-hydroxyphenyl~cetamido a^~minophe~ylacetamido .

~-ami~o~4-hydroxypheRylacatamido tri~luoro~ethylthioacet~ldo methylthioacet~ido 2,2,2 trlfluoroethyl3ulf~nyl~cet~m~do thienylaceta~ido ~-~
tetrazolyl~cetamido cyflnoacet~mi~o -~
~-carbo~tkie~ylace~mido

- 4 -~ Z~8 -a~carbo~yphenylacet~ido a-~ulfoph~nylacetamid~
me~hylsulonylacetamido ~:
cy~nom~thyl~hioacet~mido : ~ `
- a-amin~-4-carbo~ymst~yl~m~nophenylacetamido ~-amirlo-3-r'luoro-4-hydroxyphQ~ylacetarDido,' .
3-sydnonescetamldo ~
. .
4-pyrldyl~hioac~amido ~:
2 æminom~thylphenylace~mido~
~ost preferred ~ub~ti~uted ~e~razolyl groups are ~he ~110~lng:
1-3ulfomethyl~etr~olyl 1-(2-sulfoethyl)~te~razol~
1~(2-~ulamoylethyl)tetrazolyl ~ ~;
1-(3-sulfopropyl)tetrazolyl .
sul~am~ylpropyl)tetrazolyl 1-(5-~ulfopentyl~e~rszolyL
(S-sulamoylpentyl)~etr~zolyl. `~
: Partlcularly pre~0rred are the co~pounds 7-D~
:: mandelæmido-3~ 3ulfom~thyltetrazol-5-ylthiom~thyl)-3~
ceph~m-4-carboxylic ~cid, 7-D-mandelamido-3-11-(2-sulfo-ethyl)te~r~zol~5-ylthiomQthyl]-3-ceph~m-4 carboxyl~c acid, ~ ~:
7~D-m~ndelamido-3-[1-(5-~ylopentgl)~etra~ol-5-ylthiomethyl]~
3-ceph~n-4-carbox~lie acid, 7~D-ma~delamido-3~ (2-sulfamoyl~
~thyl)tetrazol-5-yl~hiome~hyl]-3-cephem-4-carbQ~Eyllc acid, 7- (2-~hierlylacetamido ) - 3- ( 1 sul fomethyl~0 ~razol- 5-y l~h~ o ~
. , m~thyl)-3-cephem-4-carboxyLic acid, 7-(2-thi~nylacetamido)~ -~
3~fl-(2~sulfoet~ e~ra~ol-S~y~thiomethyl3-30c~phem-4~
carbo~cylic acid, 7-~rifluoromethyl~h~oacetam~do-3- :
( 1- ~ulfome~hylt~trszol~5~ylt:hiometh~ 3~ceph~m-4-c~rboxyl lc acid" 7 (2-~hienylac~t~idQ3-3~ 11 (2-~ulfamoylet~l)tetra2O1-~` .

- 5 ~ - :

-lO'-~Zat~

5-ylthiomethyl]-3-cephem-4-carboxylic acid and 7-trifluoro- ;
methylthioacetamido-3-~1-(2-sulfamoylethyl)tetra~ol-5- `
ylthiomethyl]-3-cephem-4-carboxylic acid.
Cephalosporin derivatives having 7-acyl substitu-ents as defined above are all documented in the prior art.
Substitution by a substituted S-heterocyclicthiomethyl group (-CH2SHet) at the 3-position of the cephem nucleus ~ .
is also known and is disclosed in Netherlands Patent 6916151 where Het is, among others, tetrazolyl substituted with, inter alia, carboxy, carbalkoxy, alkoxyalkylamino-carbonyl and dialkylaminoalkylaminocarbonyl and in Japanese Patent 7205550 where Het includes tetrazolyl ~
substituted with -(CH2~nR3 where n is 0 to 3 and R3 ~ ~ `
includes alkoxycarbonyl, carboxy, N-alkoxyalkylcarbamoyl `
and dialkylamino. Recently issued U.S. Patent 3,819,623 ~ .
discloses cephalosporins bearing a 7-heterocyclic-acetamido or 7-heterocyclicthioalkylacetamido group and ;
having in the 3-position, inter alia, thiomethyltetrazolyl substituted with carboxy, alkoxycarbonyl, carboxyalkyl, alkoxycarbonylalkyl and dialkylaminoalkylaminocarbonyl~ ~`
alkyl. No compounds containing applicant's 3-(sulfonic ;~
acid or sulfamoyl substituted tetrazolyl)thiomethyl moiety disclosed 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 acylating agent and then displacing the 3- ; ~- -acetoxy group with the desired substituted tetrazole thiol with subsequent removal of the protective group(s). The -substituted tetrazole thiols of the formula~

lO~;'Z~:)''18 N N

N
N ~i ,-," "~
(CHR jn~S2R
''' ~:
FORMULA II~ -in which~
., i each individual Rl is hydrogen or lower alkyl;
n is one to ten; and R2 is hydroxy, amino, lo~er alkylamino or ;~
di(lower)alkylamino, are also objects of this invention, being important inter~
. .
mediates for producing pharmaceutical end products as described herein.
The carboxylic acid group of the acylating agent is activated by any of the standard methods such as con-:;: .;, , ~ersion to the mixed anhydride~ acid chloride, acid imida- ~ ~ -zolide or activated ester. In addition, a reagent such ~
."
as dicyclohexylcarbodiimide can be used provided that the carboxyl group on the cephem nucleus is protected with an easily removable protecting group such as a benzhydryl, t-butyll trichloroethyl, benzyl, benzyloxymethyl, p-nitrophenyl, p-methoxyphenyl, p-methoxybenzyl or p-nitro- ~ ;
benzyl ester. When A is NH2, the ~-amino group o the acylating agent is, preferably, protected prior to acyla-tion with an easily removable protective group known in the art such as t-butoxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl, the methyl acetoacetate adduct or similar gro~ps commonly used in the synthesis of peptides.
Alternatively, the compounds of Formula I are prepared by acylation of an appropriate 7-amino-3- ;

, :
..
"' :1.07iZOqB

substituted tetrazolylthiomethyl cephalosporin nucleus of Formula ~

S .~:;
NH2\~/

o/ \~CH2S
COOR

(C~F~ ) n--S02R
', ~',' FORMULA III ; ;
in which: ~ ;
each individual Rl is hydrogen or lower alkyl;
n is one to ten; ;~
- R is hydroxy, amino, lower alkylamino or ~` di(lower)alkylamino; and R4 is hydrogen or a protecting ester group, with an appropriate acylating agent followed by removal of the protective groups when present. ~
The compounds of E'ormula III above are also con- ;
sidered as objects of this invention.
The protective groups can be removed according to -methods well known t~ the art, such as with trifluoro-- ;
acetic 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 a basic ion exchange resin such as polystyrene-amine ion ;
- exchange resin (Amberlite IR-45~ or else by basification of an aqueous solution of the salt. ~ `
The acylating agents used as starting materials are either known or prepared by known methods.
The 7-amino-3-substituted tetrazolylthiomethyl cephalosporin starting materials of Formula III are .,~ ~, .

`: :
prepared from xeaction of 7-formamidocephalosporanic acid, prepared by reaction of 7-aminocephalosporanic acid with ;~
. . ,:
formic acid and acetic anhydride, and a substituted tetra- ~ ~;
: . ;:
zole thiol of Formula II followed by treatment with acid ;~
such as hydrochloric acid to remove the form~l group.
The substituted tetrazole thiols of Formula II
where R2 is hydroxy, lower alkylamino or di(lower)alkyl-amino are prepared by reaction of an N-alkyl dithiocarba-mate, such as methyl 2-sulfoe~hyldithiocarbamate or methyl 3-(N-t-butylsulfamoylpropyl)dithiocarbamate or its corres-ponding sodium or potassium salt with an azide such as `~
sodium azide. The n-alkyl dithiocarbamates are prepared ~-by treatment of an aminosulfonic acid, for example 2-aminoethanesulfonic acid, or an amino(N-alkyl or N,N-dialkyl)sulfonamide such as 3-aminopropane-~-t-butyl-sulfonamide or its corresponding salt with carbon disulfide and an alkyl halide such as methyl iodide in ; "
the presence of a base such as sodium or potassium hydroxide.
The amino(N-alkyl or N,N-dialkyl) sulfonamides are :
prepared by reaction of an N-alkyl or N,N-dialkylphthal-imidoalkylsulfonamide, obtained from treatment of a phthalimidoalkylsulfonyl halide, preferably chloride, with an alkyl- or dialkylamine and then with hydrazine.
The phthalimidoalkylsulfonyl halides are known or are prepared as described by Winterbottom et al., J. Amer. -Chem. Soc. 69:1393 (1947) and Griffin and EIey, J. Chem.
Soc., 3334 (1952).
When R is amino, the compounds of Formula II are .:'~' `
prepared by removal of the N-alkyl group, which also serves as an amine protective group, from the corresponding - `
~ '' '',~ ' ;:. :.
,~

1~20~8 ~ !

N-alkylsulfamoylalkyl~e~zol~-5~thiol, preferably a N-t-butylsulfamoylalkyltetrazole-5-~hiol, with, for ex~mple, ani~ole and ~rifluoroacetic ~cid~
Certain cMmpounds of th~s ~n~e~ion ar~ capable of forming sal~ wi~h, ~or example, the alkall m~tal~
such a~ sodium or potaæ~iu~, the alkaline earth m~als ~uch as calcium or with ~he am~onium cat~on. When A is ~H2, ehe compo~nd~ ca~ exis~ as the zwit~eri~n or as eith~r an acid or b~se ~alt. The~e 8alt8 are prepared by ~:
st~ndard method~ u81ng a wide ~drie~y of non-toxi~
pharmaceu~ically ~cceptable acid~ ~d ba~es known in th~
~rt and are 81go c~nsidered a~ objects of th~s invention.
It will be recognized that due to the ~symmetric :.
~; a-carbon atom ~n ~he 7-acet~mido group of Formula I when R3 is X-8H-~- and the poeenti~lly asymmetric carbon atom in : :
, .
the tetra~ole ~idecha~n, optical isomer3 wlll exi~t.
Racemic or resolved products are obtained depe~ding upon whether a racemic or resolved sidechain acid is used as an acyIa~ing agent and wh~her a racemic or re~olved tecrazole thLol i~ u~ed~ The re~olved ~ideehain acids are,readlly obtained fram tha r~cemic compounds by ;` resolution according ~o ~ell kncwn m~thods, including ; fractional cry~talllz~tio~ o~ a salt ~orm~d ~ith an opticall~ actl~e acid or ba~e. ~11 of ~he l~omer~, in-cluding separated i~o~ers and m~tures thereo~ are i~-cluded with~n the scope o~ th~ in~e~tion.
Th~ compounds of Formula T have exceptional anti bac~erial activity aga~n~t both Gr~$-positive and Gr~m- ~' negative orga~ism~. M~n~mum ~nhibitory co~centra~lons . 30 (MIC's) ranged from 0.2 to ~200 ~.tml. ~n i~ vitro ~,............... testin$. Thes~ regults are sh~wn ~ Table 1 below for , ~
'` - 10 - ~

represent~tive compo~d~ of Formula I., In vivo mouse protection dat:a 8re giv~n isa ~abl~ 2. t:ompolmd nam~s corre8pondLng to nuDbers ar- given Ln Table 3.

,'' `"

: , . :' '' ' ' ' - .'`, . 30 : ~

;; . , .~,` . `. ., 1 ' ' . ` , . ,',',.. . , ., ', . ~ , ' i ..

6~1^ . . . ~o o oo ` ~:
~u~ o~ 8n~ 0;~~ ~ :`:
~ , , .
~ ~ o o - .
L~ nnO ~ ~7 0 0 ~ _~ ~O ~ O ', ~ ~.
~ ~ ..
r~one~ ~ O . .~
VOO~ ~V ~U~~ 0 00 0 00 ;~S ~ ~ ~ o o o o o 10 8~ 33~ ~
a~ 5 0;~ ~
~ ' ' ',"~:' O
S~J
~ ~ FtlS ~ o o o ,_~ ~ oo :~ ~ .... .
.. :~
~1 9LIZI oo ~ a~ ~ ,~ a) c~ oo ~ ~.
e~l~u~BS oo ~ o ~ oo o oo , '~
' 1 5 i g ~ g o ~o o O g g ~ d8 opna~a~ ~ X ~~`I ~ X ~ ~ ~ : .
., ~ , .~: OOZI ~S ~. ~ ~ ~. ~ oo ~c~
p:l ~mn~ud q;E~I~ o o o o _- o o o o o , ~ ,'':' ~ v~ ~ ~ ~ ., o uvZ~y aa o o O ~ ~ _~ o ,~ o o - "
2() ~u~ q~T~
~ ~ ' .
6L~ 1~ ~1 ~ .~ .~ ....
O~IZI ~ ~1;~ El oo o o ~ ~

:l 8S~q6 N~ ~ o ~ o u~o o oo 2 5 ~ d~S ~ u~
- . . , znlsT~ S ~ID o ~ ~ s~ ~ ~ ~ o ~
~: -. ~
~r i 06~1E Z ~XS 8na~n~ ~ So ~o o o o o o o o o :-'' , ~_--_ .ZI UH ~n~n~ S ~ ~ ~ --' ! ~ : :
.~ p~d~o ) ~ , P P
~ .
:j :
. . .
, - , !

5:' ~78 `''~

o o ~- .
o o oo oD ' ~ ~
o o . .

o o o o o ~
~ ;-,, ' ~ ' , "
~ o ~/.
.
.. .
: , 1 5' o C ~
o o "~ . .
~;t `~ '' "
. ~. .
~o .

~ o .
~ 20 ~ . -, ~ ~ ~

:. ao oO "
, . . .
~. oo :

:` _ . ~, .
," ~. U~ '.:

~, ' S _ . :
., . . .: . ~
:. ~1 ~D .
: J
o o : '.

o ,.
~`,3 3rJ ~ _ ,1, ~ ~i'-''.`'` .
.,. ,~, .
-3 ~ ~i .,, . . -, ' ' `' ' .` 1 3 -- ' -1(~'7Z~

TABI E
~ 50 in ~iVQ (mg.lkg.) Compound E. coli 12l40 ~ ~C~
~ O c . ,~. ~ O c . ,~"
1 . 56 -- 0 ~ 28 ~1û ,. 58 35 Q . 3918 0 6 III0.70 25 0.68 2105 . ~:
.
IV 18 ~50 150 7 --Y 4 ~50 ID ! 2 4 __ ~,5 ` 10 , "
~tII0. 86 -- 0 . 24 - ~
VIII . l . 32 >50 2 _~ :
IX 1. 56 44 1 5 ` :

..

:. I 7-D-Mand~Lamido 3~ sulome~hyl~
te~razol-5-ylthiom~thyl)-3-cephem- `
~` 4-c~rboxylic acid . ~ ~ 20 II 7-D~d~lamldo-3- ~1- (2-sulfoethyl) -te~razol~S-ylthlomethyl 1 -3-cephem- .
; 4-carboxyl lc ac id ~ III 7-D-~Mandelamido-3-[1-(2-sulfamoyl-. ethyl)tetrazol-5-ylthi~m~thyl]- ~, : 3-ceph~m-4-carboxylic acid :
:~. IV : 7-D-Mand~lamido-3-El-(5~ulopentyl)~
. 2 5 tetr~zol-S-ylthiomethyl~ -3-cephiem- : 4-carbox~l ic Elc id - :, V 7-(~-Thlenylacetamldo)-3-~l-sulfo- : ~
me~hylte~crazol-5-ylthiomethyl)-3- `- :
cephem-4-carbo~cylie acid . ~ VI 7- (2-l~iertylaeet~mido) -3~ (2-sulfo~thYl~ tetra~ol5- ylthio~
methyl]-~-~ephem-4~carboxylic acid :~
.- 30 VII 7-TrifluoromethYlthioacstamiido-3~ sul~o~eithyit~trazol-5~ylthio-hyl3-3-cephei~-4-~rboxylic acid :` :
.

'' ~,, VIII 7- (2-Thienylacet~mldo)-3~ 2~gulf3moyl~
ethyl)'ce~r~zol-S~ylthiom~thyl]-3~cephem-4- ~:
carbo:Kyl ic ~c id IX 7-TrifluoromethyL~hioaceta~Qi.do-3 ~1- (2-~ulfamoyl~thyl)tetrazol-5~ylthio~
methyl l - 3-cephem~4~carboacyl ic ar id In addition, the active c~mpou~d~ of this in~ren~
tion exhiblt bro~d Rpectrum ac~iv~ty ~nd ~how advan~ageou~
ly high blood $erum levels ~d hal~-llfe values.
Pharmaceutical compositi~n~ havin3 a~ntibacterial activil:y whLch c~mpri~e a phsrmaceutical carrier con-tainix~g an act~e but non-~ox~ qu~tity o~ a compound - ~:
:
of l~ormulP I as well as m~chod~ of combatting bac~erial ~iEections by ad~n~ni~tering such a composition tO an infected hogt i~ a nonto$ic am~unt ~ufficien~ to combat such LTLfections are al~o ob3ects o~ this invention.
The adm~nistration may be by parenteral injection such as ~:
subcutaneously, intramut3cularly or intravenously. The ~-J ~ . in~ection of su~:tably prepar~d ~terile solu~cion~ or i' ~ suspensions contal~ g ~n efec~ive, nonto~cle am~unt of .' 20 - the ne~ cephalo~porin compou~d i8 the pre~erred route of ,ij . .. .
. administration.
The compouDd~ o~ Fon~ula I are formul~ed and :
administered in the same manner a~ other cephalo~porins.
~, The dosage regimen comprise~3 admini~trat:ion, pr~ferably -, 25 by in~ection, of an ~ctive but Yaon~oxic quantity of a ~ -compound of Fbrmula I select~d from the dosage unit range of from 100 to lOOû m~. with the total daily dogage ` ^, -~ regimen being rom 400 mg. to 6 g. 'l~he preci~e do~age~
.. ,~ are depeT~dent upon the age and weigh~ o ~he subject -~t 30 and on ~he in~c:tion being treated ~d can be det~ ned ~:
by those skilled ln ~he ar~ b~ed OTI the da~a disclosed ' '; ~.,,~:
- 15 ~ .

q~ ::

herein c~mpared with that ~vailable to the ~t attained with known cephalo~porlns.
The following e~:~mples illustrate the invention, , ~ :
but are not to be oons~rued as limit~ng the ~cope thereofO
Temperature~ ar~ ~n degrees C~ntigr~de unles~ otherwise :`
statedO
EXAMn~ 1 "~

-ce~ c~r o~ I I c ~c :
To a ~olutic~n of 112 ~. (2.0 Dlol.) of potassium ~:
hydroxide arld 111 g. (1.0 mol.~ of aminome1:han~ulfonic : .
acid in 250 ml. of water at 25 wa~ add~d 71 ml. of carbon ~`~
disulfide0 The reaction mixture wa6 ~tirred for 12 hour~
and 250 ml. of ethanol was addad, The reaction ves3el was fitted with a reflusc.c~denser and 62 ml. (l.0 mol.) of methyl iodLde wa~ ad~sd. When the exothermic reaction cooled to anbient t0mperature the ~olid produc~ was collected by filtrstionO The solid was ex'cracted with ;~, hot methanol and the extract wa~ concentrated to give ;, me~hyl ~ulomethyldi~hiocarb~mate ~18 the potas~ium Balt~
v~l A mixture of 45.3 g. (0.19 mol.) of methyl sulfo-methyldithio~arbamate pota~lum ~alt a~d 16 . ~ g . ~0. 26 mol.) of sodium azide in 425 ml. of water was heated :~ at 80 for 4.75 hours. The reac~ion mixture was pa~ed 2~ through an AMberlite IR-120H ~on excha~ge reAin column and eluted w~th water ~n~iL the pH of the eluant became 3.5. The eluan~ wa~ extrac ed wl~h e~he~ and the aqueous ~olution wa8 evaporated to dryness to gi~e l-~ulfo-methyltetrazole-5~th~ol~
~0 l~Sulfone~hyltetra~ole-5~hiol wa~ dissolved in acetone and a 30% ~olution of ~odium ~e~hylhexanoate ~ .

~ 16 -z~
''.

in isopropanol wa~ added. l-Sulfomethyltetrazole-5-th~o~
sodium salt precipit~ted snd wa~ collected by filtratlon. ;~
A mixture of 27.4 g. (0.062 mol.) of 7~D-mandel-- 5 amidocephalosporanic acid meth~nolate, 10.2 g. (0.047 moi.) ~-o~ l-sulomethyl~trazole-S- hlol so~lum s~lt and 9 . 2 g.
~0~109 mol.) of sodium bicarbonate ~ 300 ml. of water was heated at 70 for one hour. The reaction mixture -was cooled (lce bath) and acldified ~o pH 1.8 with 3N ~:
hydrochloric acld. The mixture was ex~raoted wlth ethyl ~ ~ -~ceta~e, ~ fil'cered and chrotna~ographed on a~ Amberlite XAD-8 resin coJumn wi~h w~cer conta~ing increasing amounts of methaslol a8 ~he eluan~ to give th~ title compound .
. 15 The tltle coimpound was di~olvet in methanol ~nd , ~ 5~ solution of sodium methoxide ~n methanol was added :~ uneil pH 700. Addit~on of e~hanol precipiSated the ~ .
,.j .`~; produc~ salt which was collected, dissolved in water a~d ~.
.. ~ lyophilized ~co give 7 D-mandelsmLdo-3~ sulfomethyl- ~:
~: 20 tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid ~i' disodium:aalt. `,~ :
l C18Hl~N68$3 2 Na 1-75 H20 Calculated: 34.98b C, 3.18% H; 13.$9% N
Found: 35.09% C~ 3.17% H; 13,27% N
2S EXA~PLE 2 ... .

.:! A solutio~ of 7.58 g. ~0~015 mol~) of 7-(D a~
t-butoxycarbonylam~nophenylacetamido)cephalosporanic acld, 1.96 g. (0.01 mol.) of l~sulo~hyl~etr~zole-5~thiol and ~ .
'':
'' l~ q~ "`' ;:' :' 2.52 g. (0O03 m~l.) of sodium bicsrbonate ln 125 mL. of water is stirred at 60 ~or five hours while maintaining the pH at 7.0-7.~ by addition of sodium bicarbonate. The .
mi~ture is cooled a~d e~r~c~ed with e~hy~ acetate~ The ~queous phase 1~ acidi1ed to pH 2.5 wlth 3N hydrochlorLc acid ~nd ~he acidic solution ls extrac~ed aga~n w~th e~hyl ace~a~e, Thg aqueou~ ph~e i~ brought to p~ 7.1 Dy additi~n of 5% ~odium c~rb~nat~ solution, then passed through a XAD-4 ion e~çhange resin column and eluted with :
-water and methanol to g~ve 7~(D~a-t-butoxycarbonylamino-phenylacetamido)~ sulfomethyltetrazol-5-ylthiomethyl)- ~
3~cephem^4-carboxylic ~cid disodium ~alt. r 1S 7-(V-a-t-butoxycarb~nylamlnophenylacetamido)-3- -i (l-sul~omethyltetrazol~5-ylthiomethyl~-3-cephem-4-carboxylic a~id disodium salt is ~tirre;d at 25 with 25 ml, of ; -~
trifluoroacetic~ acid and 25 ml. of 1,3-d~ thoxybenzene ; .
for 2~25 hours. The mix,ure i~ evaporated to dryne~s, ether is added to the re~idue and the precipltate is collec~ed, washed with ether~ stirred in acet~nitrile .
for two hours, then collected and dri~d in vacuo to give :~
the tit~e compound.

. 25 A soluti~n of 2.73 g. (0.01 mol.3 o~ 2-, ~

~;

- 18 - .

~ 8 -phthalimidoethanesulfonyl chloride in 20 ml. of chloroform was added dropwise to a solution o 2.19 g. (0.03 mol.) -of t-butylamine in 20 ml. of chloroform at 5. The reaction mixture was warmed to ambient temperature and stirred for three hours. The precipitate was removed by filtration and the filtrate was evaporated to dryness to give a residue which was purified by chromatography on silica with l9ol chloroform-methanol as eluant to give 2-N-t-butylphthalimidoethanesulfonamide.
..
2-N-t-Butylphthalimidoethanesulfonamide (2.10 g., ~;
6.78 mmol.) was suspended in 20 ml. of ethanol and 0.344 g.
of hydrazine hydrate was added. The reaction mixture was refluxed for three hours, then evaporated to dryness.
The residue was suspended in 45 ml. of water and acidified ~ ~
to pH 3.0 by addition of dilute hydrochloric acid. The `~ ;
acid solution was filtered and the filtrate evaporated to dryness to give 2-aminoethane-N-t-butylsulfonamide -hydrochloride.
2-Aminoethane-N-t-butylsulfonamide hydrochloride (1.25 g., 5.78 mmol.) was added to a solution of 1.17 g. ;;
(11.56 mmol.) of triethylamine in 20 ml. of ethanol.
Carbon disulfide (0.44 g., 5.78 mmol.) was added, the mixture was stirred at 25 for 1.5 hours, then 0.82 g. `
(5.78 mmol.) of methyl iodide in 5 ml. of ethanoL was ;
added and the resulting mixture was stirred for 1.5 hours. ~ ~;
The mixture was evaporated to dryness and the residue was dissolved in water and acidified to pH 2.0 with dilute hydrochloric acid. The aqueous mixture was extracted with ethyl acetate and the extract was dried (MgSO4) and ~-evaporated to dryness to give methyl 2-(N-t-butylsulfamoyl)-ethyldithiocarbamate.
` '~ .

. ~..
,,~ , Methyl 2-(N-t~butylsulfamoyl)ethyldithiocarbamate was treated with sodium azide as described in the procedure ~-of Example 1 for 35 minutes to give 1-(2-N-t-butylsulfamoyl-ethyl)tetrazole-5-thiol.
l-t2-N-t~Butylsulfamoylethyl)tetrazole-5-thiol (1.0 g.) was suspended in 10 ml. of anisole and 20 ml.
of trifluoroacetic acid is added. The solution was heated at 56 for 3.5 hours, then cooled. The precipitate was ~
collected by filtration and washed with petroleum ether ~s to give 1 (2-sulfamoylethyl)tetrazole-5-thiol. ;
A solution of 0.210 g. (2.5 mmol.~ of sodium bicarbonate in 5 ml. of water was added to a suspension -of 0.272 g. (1 mmol.~ of 7-aminocephalosporanic acid in 5 ml. of water and 2.5 ml. of acetone at 15. The ;~
solutionwas heated to 45, a solution of 0.314 g. (1.5 mmol.) of 1-(2-sulfamoylethyl)tetrazole-5-thiol in 10 ml.
of acetone was added and the reaction mixture was refluxed for two hours~while maintaining the pEI at 7.4-7.6 by ~;~
:, I . :.
~ additIon of aqueous sodium bicarbonate solution. The `~l 20 mixture was cooled and acidified to pH 4.0 with dilute hydrochloric acid. The precipitate was collected by `
filtration to give 7-amino-3-[1-(2-sulfamoylethyl)tetrazol- -~
5-ylthiomethyl]-3-cephem-4-carboxylic acid.
To a solution of 1.26 g. (15 mmol.) of sodium bicarbonate in 75 ml. of acetone and 50 ml. of water at , 5 was added 2.1 g. (S mmol.) of 7-amino~3-[1-(2-sulfamoyl-ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid. The solution was cooled to -10 and a solution of 1.55 g. (5.5 mmol.) of D-0-dichloroacetylmandeloyl chloride in 25 ml. of acetone was added. The reaction mixture was stirred for 30 minutes in the cold while '`~' ~' ~ ~ - 20 -~3~
,, :
maintaining the pH at 7~2 by addition of aqueous sodium ~;
bicarbonate, then for 1.5 hours at 25. The mixture was extracted with ether and the aqueous phase was brought ;~
to pH 9.3 with 5% sodium carbonate and stirred for 1.5 hours at 25. The aqueous mixture was extracted with ether, the pH adjusted to 4.5 and the solution was again extracted with ether. The aqueous phase was acidified to pH 1.5 with dilute hydrochloric acid and ~ ; it was extracted with ethyl acetate. Evaporation of the j 10 extract to dryness gave a solid which was suspended in ~-ethyl acetate and filtered. The filtrate was diluted with ether and petroleum ether to precipitate the title j compound.
', EXAMPLE 4 ;

, 7-D-Mandelamido-3-[1-(2-N-t-butylsulfamoylethyl)tetrazol-5- `
l ylthiomethyl]-3-cephem-4-carbox i c acid ~;~
1~ When an equivalent amount of 1-(2-N-t-butyl-Il sulfamoylethyl)tetrazole-5-thiol is reacted with 7-amino- `;~
i~ cephalosporanic acid as described in Example 3, 7Aamino-3- ~`
[1-(2-N-t-butylsulamoylethyl)tetraæol-5-ylthiomethyl]-3- , cephem-4-carboxylic acid is obtained. ~;
Acylation of 7-amino-3-[1-(2-N-t-butylsulfamoyl-ethyl)tetrazol-5-ylthiomethyl]-3-cephem-~-carboxylic acid with D-O-dichloroacetylmandeloyl chloride according to the procedure of Example 3 gives the title compound.

7-D-Mandelamido-3-[1-(2-N-methylsulfamoylethyl)tetrazol-5-lthiomethyl]-3-cephem-4-carboxylic acid Use of methylamine in the reaction with 2-phthal-imidoethanesulfonyl chloride described in Example 3, followed by the subsequent synthetic steps described `
therein gives l-(2-N-methylsulfamoylethyl)tetrazole-5-thiol.
.. ''`~ ~`'`

' ~,', Z0~8 ~. "

When 1-(2-N-methylsulfamoylethyl)tetrazole-5-thiol is reacted with 7-aminocephalosporanic acid as described in Example 3 and the product 7-amino-3-[l-(2-N-methylsulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid is acylated with D-O-dichloroacetyl-mandeloyl chloride as described therein, the title com-pound is obtained. ;~ r ` EX~MPLE 6 7-(D-a-Amino-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoyl-ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid ;;
A solution of 7.82 g. (0.015 mol.) of 7-(D-~-t-butoxycarbonyl-4-hydroxyphenylacetamido)cephalosporanic acid, 4.6 g. (0.022 mol.) of 1-(2-sulfamoylethyl~tetrazole-5-thiol and sodium bicarbonate are reacted according to ., ; . .
; the procedure described in Example 2. After cooling, the ;l reaction mixture is extracted with~ethyl acetate. Fresh . ;. , ;
-/ ethyl acetate is added to the aqueous phase and it is -~
:., : ..
acidified with stirring to pH 2~8 with 6N sulfuric acid.
The layers are separated and the aqueous phase is again extracted with ethyl acetate. The combined extracts are washed with water, dried (MgSO4) and the solvent evaporated to give 7-(D-a-t-butoxycarbonyl-4-hydroxyphenylacetamido)-3-[l-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid. ~-7-(D-~-t-Butoxycarbonyl-4-hydroxyphenylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-S-ylthiomethyl]-3-cephem-4-carboxylic acid is treated with trif~uoroacetic acid as ;~
described in Example 2 to give the title compound.~i ;

7-D Mandelamldo-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]--- ;
3-cephem-4-carboxylic acid 2-Aminoethanesulfonic acid (50 g., 0.4 mol.) was - - 2 ~
,`; :

.- . .. .- . . . - .. ... . .

7Z0~8 ~ ~ ~

added to a solution of 45 g. (0~8 mol.) of potassium hydroxide in 100 ml. of water at 25. Carbon disulfide (24.4 ml., 0.4 mol.) was added and the reaction mixture was refluxed for 2.5 hours. Ethanol was added to the warm solution, the mixture was cooled to ambient tempera~
~. . ..
ture, 57 g. (0.4 mol.) of methyl iodide was added and the resulting mixture was stirred for 1.5 hours~ The -mixture was evaporated ln vacuo and the residue recrystal-`~ lized from hot ethanol containing 3~ water to give methyl ~',tl 10 2-sulfoethyldithiocarbamate potassium salt.
A mixture of 21.5 g. (0.087 mol.) of methyl 2-sulfoethyldithiocarbamate potassium salt (0.5 hydrate) and 7.16 g. (0.11 mol.) of sodium azide in 200 ml. of `
. ~! ' ', water was refluxed for two hours. The solution was cooled ~ ~
to 25 and extracted with ethyl acetate. The aqueous ~`
phase was treated with Amberlite IR-120H resin, washed -~
with ether and evaporated to give an oil. The oil was `~-dissolved in acetone, the solution was filtered and the filtrate was evaporated to dryness to give 1-(2-sulfoethyl~
tetrazole-5-thiol. The thiol was dissolved in isopropanol, cyclohexylamine was added until pH 8-9 and acetonitrile was added to give 1-(2-sulfoethyl)tetrazole-5-thiol as the di-cyclohexylamine salt.
.:
1-(2-Sulfoethyl)tetrazole-5-thiol di-cyclohexyl-amine salt was dissolved in water and treated with Amberlite IR-120H resin to give 1-(2-sulfoethyl)tetrazole- `
5~thiol. -~
To a solution of 2.1 g. (0.01 mol.) of 1-(2- -' sulfoethyl)tetrazole-5-thiol in 100 ml. of water was added 6.4 g. (0.015 mol.) of 7-D-mandelamidocephalosporanic acid ~, sodium salt and 1.68 g. (0.02 mol.) of sodium bicarbonate.
` , The mixture was stirred at 70 for 2.5 hours then cooled and acidified to pH 1.8 with 3N hydrochloric acid. The ~ ;
acid solution was extracted with ethyl acetate and ether, acidified to pH 0.9 and chromatographed on a XAD-8 resin column with water as eluant to give the title compound.
The title compound was converted to the correspond-ing disodium salt by treatment~with sodium methoxide as described in the procedure of Example 1.

19 18N6O8S3 2 Na 2 H2O
Calculated: 35.85% C; 3.48~ H; 13.20% N
Found: 36.21% C; 3.29% H; 12.96% N

i; .
7-D-Mandelamido-3-(l-sulfamovlmethvltetra2o1-5-ylthiomethYl)-., ~ .
, 3-cephem-4-carboxylic acid ~
:: .
A suspension of 15.1 g. (0.136 mol.) of amino-methanesulfonic acid and 14.2 g. (0.145 mol.) of anhydrous ~ ;
'~ potassium acetate in 48 ml. of acetic acid is refluxed for - ten minutes. Phthalic anhydride (21.4 g., 0.145 mol.) is ; then added and~the resulting mixture is refluxed for 2.5 hours. The product is collected by filtration and washed with acetic acid and ethanol to give phthalimido~ ~
methanesulfonic acid potassium salt. `
To 41.7 g. (0.15 mol.) of phthalimidomethanesul-fonic acid potassium salt in 220 ml. of dry benzene is `
added 22.5 g. (0.132 mol.) of phosphorus~ pentachloride.
; The reaction mixture is refluxed on a steam bath for one ~;
hour, then an additional 22.5 g. of phosphorus pentachloride ' ~; is added and heating is continued for 1.5 hours. The reaction mixture is evaporated to dryness, crushed ice is added to the residue and the slurry is filtered. The ; product is washed with water to give phthalimidomethane- ; ~
~: ' - 24 - `
` `' '',.':
, .' ~ 78 '';' ' sulonyl chlorid~.
When phthalirnidomethan~sulfonyl chloride is sub- :;
stituted in th~ procedure o~ Exampl~ 3 or 2-phthal~mido- ~:
ethanQsulfonyl chloride, N-t-butylphthalimidomethanesul~on-amide i9 prepared which is converted to l-N-t-butylsulfamoyl~
aethylte~razole-5-thiol as d~cribed therein. Trea~cment of l-N-~-bu~ylsulfamoylme~hyltetra~ole-5-thiol with ~rifluoroac~ic acld as desc~bed in Example 3 gives sulfamoylmel~ylt~trazole-5-~hiol, ~`: 10 .. ::
. ~eactLon of l-sulfau~oylmethyl~ce~razole-5-thiol with 7-aminocephalo6poranic acid and treatment of the ;:
: product 7~ o-3-(1-sul~noylmethyltetrazol-5-ylthio-methyl)-3-cephem-4-carboxylLc acid with D-O-dichloro~
~!, acetylmandeloyl chlorlde as described in ~xample 3 gives j 15 ..
the ti~le compound. .

:~ su Y ethv )tetrazo ~ -v t t Y ~ -ceP em- -car oxy ic , .

2-N,N~Dimet~ylsulfamvylethyl)tetrazole-5-thiol .~0 is prepared from reactLon of dimethylamine and 2-phthalimido-etha~esulfonyl chloride followed by eonv~rsion of the product thu~ obte.ined to the tetra~ole thiol by the react ion ~ ~.
se~uenoe de~crlbed in the procedure o~ Example 3. ~ :~
i ~hen 1-(2-~,N~d~methylxulfamoylethyl)tetrazoleo5 ~ 2 :~ thiol is reacted with 7-(D~a-t-butoxycarbonyl-4-hy~roxy-:, phenylacetamido)cephalosporanic acid as de~cribed in ., Example 6 a~d the product is de-blocked as described above, . .
the title compou~d is ob~ained.
EX.~,E 10 : 30 7-D-Mandelainido-~-[1~3-sul~oDroDvl)tetrazol-5-vlthlo- :

When an ~quiv~lent am~unt of 3-aminopropane-, : :. .

` - 25 - .:
.... .. . . . . .. .

Z~7~ -sulfonic acid was ~ubs~ituted in the procedure of Example 7 for 2-~mino~than~sul~onic ~cid, ~thyl 3-~ulfopropyldithio-carbamat~ pota~slum ~al.~ was prepared.
Reaction of methyl 3-~ul~opropyldithiocarba~te potas~ium ~alt with sod~um a~îde 8S described ~n Ex~mple 7 ~:
gave 1- (3-suïfopropyl)~etrazol~-5-~hlol .
Substi~u~ion of an e~uivalent ~mount of 1-~3-~ul~c3propyl3te~azole-5-~hiol in ~h~ procedure o~
, ~. Example 7 in plac:e of 1- (2-~ulfo~thyl) ~e~razole-5-thiol ~ 10 in the re~ctlon with 7~ ndelsmid4cephalo~poranic acid sodium ~sLt g~ve~ the title compound. ~ ~:
The title compound is converted to the corresponding sodium salt as d~scrlbed in th~ procedure of Ex~mple 1.

Wher~ 3- phth~1imidopropanesulfonyl chloride i~
- substituted in ~che procedure of ~ample 3 for 2-phthalimido~
ethanesulfonyl chloride, 3-~-t-bu~ylphthalimidopropane-sulfonamide i~ prepared, which i~ co~verted to 1-(3-~-t~
butylsulfamoylpropyl)te~razole-5-thiol as deseribed therein. Treatment vf 1-(3-~-t-b~tylsulam~ylpropyl)-tetrazole-5-thiol with trl1uoroacet~c acid 83 described in Example 3 gLves 1~(3~sulfamoylpropyl)tetrazole-5-thiol.
aeactlon of 1-(3 ~lfaylpropyl)tetrazole-5-thiol with 7-aminocephalospor~nic acid and treatment of the resulting 7-amlno-3-[1-(3-sulfamoylpropyl~tetrazol-5-ylthiomethyL]-3-cephem~4-oarbo~cylic aeid wl~h D-O-dichloro-acetylmandeloyl chlo~ide a~ described iTI ~ample 3 glves the t it le compound, ~ 8 EXAMPLE 12 ~ ,~
7-D-Mandelamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthio-, methyl]-3-cephem-4-carboxylic acid : Substitution of an equivalent amount of 5-amino-`, pentanesulfonic acid in the procedure of Example 7 for .
2-aminoethanesulfonic acid gave methyl 5-sulfopentyl- '.:.,'., dithiocarbamate potassium salt. "~ ,~
,, Reaction of methyl 5-sulfopentyldithiocarbamate ,~
,~ potassium salt with sodium azide as described in Example ~ 10 7 gave 1-(5-sulfopentyl~tetrazole-5-thiol. ,~
-~, A mixture of 4.55 g. (0.010 mol.) of 7-D-mandel- ,~
,, amidocephalosporanic acid sodium salt and 3.25 g. (0.011 .:~
mol.) of 1-(5-sulfopentyl)tetrazole-5-thiol disodium salt, :, prepared as previously described, in 80 ml. of water at `;~' ,' pH 7.2 (adjusted by addition of sodium bicarbonate) was ,"' :
heated at 65 for 4.5 hours. The reaction mixture was , ~
cooled,:~.acidified to pH 1.6 with 3N hydrochloric acid '.'' '' and extracted with ethyl acetate. The pH of the aqueous .',''.
phase was brought to 7.0 by addition of sodium bicarbonate and the solution was chromatographed on a XAD-4 resin ;'~
column eluting with water and then methanol. The product was dissolved in methanol and ethanol was added to the ,';~
solution to precipitate the title compound as its di-sodium salt. ~ , ` C22H24N6853 2 Na 1.5 H2O 0.75 C2H6O .:
Calculated: 40.08% C; 4.50% H; 11.93% N; 13.65% S `.
: Found: 40.01% C; 4.20% H; 10.71% N; ].3.54% S ..
~, 7-D-Mandelamido-3-[1-~5-sulfopentyl)tetrazol-5- :''' ylthiomethyl],-3-cephem-4-carboxylic acid disodium salt '~
is converted to the title compound~:as described herein.

' - 27 ~

'' 10'72~7~
... .. .. .
EXAMPLE 13 ~ ~
~. 1 7-D-Mandelamido-3-[1-(5-sulfamoylpentyl')tetrazol-5-ylthio-methyl]-3-cephem-4-carboxylic acid '~
- Use of 5-aminopentanesulfonic acid in the pro-, cedure of Example 8 in place of aminomethanesulfonic acid ' , ''~ followed by reaction of the product thus obtained with `~` phosphorus pentachloride gives 5-phthalimidopentane- '-,~
, - .
';~ sulfonyl chloride. '~ ~
~, When 5-phthalimidopentanesulonyl chloride is '' ~;
, lO used as a starting material in the sequeDce described in Example 3, 1-(5-N-t-butylsulfamoylpentyl)tetrazole-5-thiol `,' ,'' is obtained. Treatment of 1-(5-N-t-butylsulfamoylpentyl)-. ,,. "
-, tetrazole-5-thiol with trifluoroacetic acid as described ,, above gives l-(5-sulfamoylpentyl)tetrazole-5-thiol. ~,;
Reaction of 1-(5-sulfamoylpentyl)tetrazole-5-,, thiol with 7-aminocephalosporanic acid and treatment ' of the resulting 7-amino-3-[l-(5-sulfamoylpentyl)tetrazol~
'.-: ~ .
'~ 5-ylthiomethyl]-3-cephem-4-carboxylic acid with D-0- ,-dichloroacetyLmandeloyl chloride as described in Example 3 gives the title compound.

7-Trifluoromethylthioacetam do-3-tl-sulfomethyl- ~,',' ~etrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid A solution of 2.18 g. (10.0 mmol.) of l-sulfomethyl- ' ' tetrazole-5-thiol sodium salt, prepared as described -, above, 0.840 g. of sodium bicarbonate and 5.45 g. (12.5 ~' mmol.) of 7-trifluoromethylthioacetamidocephalosporanic ::.
acid sodium salt in 60 ml. of water was stirred at 70-75 `','~
, for five hours while maintaining the pH at 6.8 by ' ~' addition of 5~ aqueous sodium carbonate solution. The ;~;, ; reaction mixture was cooled and diluted with water. i,~
Ethyl acetate was added and the mixture was acidified ~. , , ~.'.

~', '" .
., ~ 7~ ~
, .
to pH 2.0 with 6N hydrochloric acid. The aqueous phase `
was extracted with ethyl acetate then brought to pH 6.8 ;~
by addition of sodium bicarbonate and chromatographed on ;
XAD-4 resin with water and methanol as eluants. The product-containing fractions were evaporated to dryness to give a residue which was dissolved in water and lyophilized to give the title compound as the disodium salt.

13~11F3N67S4 2 Na 2.25 H20 ~`
Calculated: 24.59~ C; 2.46~ H; 13.23% N `~ ;~
Found: 24.54% C; 2.18~ H; 12.85% N
, . .
An aqueous solution of 7-trifluoromethyl thio acetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid disoaium salt is treated with Amberlite IR-120~ ion exchange resin as described above`~ `~
to give -the title compound. `
E~AMPLE 15 7-Trifluoromethylthloacetamido-3-[1-(3-sulfamoyl- ;
propyl)tetrazol-5-ylthiomethyI]-3-cephem-4-carboxylic acid 7-Trifluoromethylthioacetamido-3-[1-(3- ~ `
sulfamoylpropyl)tetrazol-5-ylthiomethyl~-3-cephem-4-carboxylic acid sodium salt is prepared by substitution -~ of an equivalent amount of 1-(3 sulfamoylpropyl)tetrazole-5-thiol sodium salt, prepared as described above, in the procedure of Example 14.
The title compound is obtained from the salt as described above.
EXAMPLE 1~; -^
7-Trifluoromethylthioacetamido-3-~1-(2-sulfamoylethyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid A mixture of~2.18 g. (5 mmol.) of 7-trifluoro-;:

lO~Z~

methyl~ioae~ta~idocephalosp~r~nl.c acid ~od~ alt ~
~d 1.75 g. (8 D~l.) o~ 1-(2~ul~oylethyl)~etraæole-5- ~;
thiol ~n 50 ~1. of wEter m~i~t~ d at p~ 7.~ by addition of sodium lblc~lrbonate W~8 hea.ed at ~0-70~ or three hours. The r~action mixture waa eooled, acidiied ~o pH 3.5 and eD:tralc~ed with ethyl ac~t~te~ The ~trsct wa~ w~hed with ~7s~er, dried (~04) ~nd ~vaporated to dryness to give a re~idue ~hich ~ dl~olved ~n ~hyl ~ .
acet~t~" ~th~r wa~ added, th~ ~olutiogl wa~ filtered, . . .~ .:
p~troleum ether w~ added to the iE~ltrate and the re~ul~ng prec~pitat~ wa~ coll~cted by flltrat$on a~d dis~olved ln m~thanol. A 5% solutio~ of eodium methoxlde ~n ~th~ol wa~ added to th~ m~thanol solution ~ .
un~ll pH 7.1. Eth0r ~a~ dd~d ~nd ~he procipitate was collected and dried in vacuo ~o giv~ 7-trifluorom~thyl- : :
thiosc~a~ido - 3~ (2-~ulf3moylethyl)tetrazol-S-ylthiom~thyl]-3~c~ph~m-4-carboxylic acid sodium ~alt. :~
7-Trifluoro~ethylthioac~tamld~-3~ t2-~ulf~m ethyl)te~razol-5-ylthio$0thyl]-3-cephem-4-carboxylic acid sodium salt i~ co~erted to the ~ co~pound by : m~thods de~cribed ~bove.
EXAMPL~ 17 ~ ~, A ~uspension o 56 g. ~1.0 ~cl.) of pota~slum hydroxide and 118.7 g. ~0.5 m~ o 10-a~inodec~ne-sulf~c acid ~n 170 ml. of ~ater i5 ~tirred for 30 :
mi~ute~ at 25 ~hen 40 g. ~0.52 mol.~ of c~bon di~ulfide and 80 ml~ of e~hanol are added a~d the re~c~ion mixture i~ ~tirred at 25 for ~2 hours. The mix~ure i~ re1uxed gently for tw~ hour~ and cooled. ~ethyl iodide (71 g., `.

- 3~ -'7X~7 0.3 mol.) and 130 ml. of ~th~ol are added to th~ mlxtu~
a~d it i3 ~ d at 25 for 12 hours. The allix~cure i8 ;
evaporat~d to re~e the eth~nol ~nt the ~olid re~i~ue i8 eollected by filtr~tio1n to ~iY~ thyl 10-~ul~d~cyl-dithioc~rb~t~.
!~thyl lO~ulfo~ecyldithiocarb~aSe (31.4 g., 0.096 m~ r~acted with 6.5 g. ~0.,1 m~) of ~diu~n ~zide a~ cr~b~d ~b~v~ to giVQ l~(lO~aulot~cyl)-tetrazole-5-th~ol.
1~10-Sul~od~cyl)tetr~ole~-5 thiol (4.84 g., 15 ~ol.) is ~lowly ~dded bD a ~ tion of 3.36 g. : :
(40 m~nol.) of sod~ b~car~ate i~ 100 ml. of water.
7-D-~andel~midocephalosporan~c acid (4.20 gc, lO n~nol.) is then added ~d the mixturo i5 h~at~d at 65 for 3.5 hour~. The ~ure i8 filtered, the filtrate i8 ex-tracteâ with ethyl sc~tate and the aqueou~ layer i~
acidified ~co p~ 4 ant eac~cract~d agai~ wi h e~hyl ace~ate.
The e;ctr~ct i~ drie~d ~M~S~) arld evap~rated to dryness t:o giV8 ~I~B title coDapouE~d.

U8~ of lQ^~lnodec~n~sul~onic acid in the proc~sdure of Ex~mple 8 in place of ~ino~thanesulfonlc acld ~ollowed by reaction of. the product ~hua ob~ain~d with phosphorus pentachloride give3 10--plhthalimidodecarle~ulfonyl chloride.
~en 10-phthalimidodlocane~ulonyl chloride i~
~: u~ed a8 a startiL~ ~at~r~al ln the ~uence de~cribed ~n }3x~mple 3, 1-(10 N~ butyl~ulfa~oyldacyl)tetrazole S- :
0 thiol i~ obta~ned, Tre~t~ent of l-(lO~N-t-bu~ylsulf~moyl-decyl)t~tra~ole-5-t~lol ~ith ~rifluo~oace~ic ~cid glv~s .:

: - 3I - :

~ , , . ~ ., , . ,, . , ... ~

(10-sulfamoyldecyl)tetrazole-5-thioL.
React~on of l-(10-sulfamoyldecyl)tetrazole-5-thiol with 7~aminocephalosporanic acid and treatment of the resulting 7-amino-3-[1-(10-sul~amoyldccyl)tetrazol-5-ylthiomethyl]-3-cephem-4-sarboxylic acid with D-0-dichloro-acetylmaIIdeloyl chloride as described in Example 3 gives the title compound. ;
EXAMPLE 19 ~ ~

~' - `. "
~ Substitution of an equi~alent amount of 2-amino-; propanesulfonic acid in the procedure of Exampl~ 7 for . 2-amlnoethanesulfonic acid gi~es methyl (2-sulfo-1-methyl)-ethyldithiocarbamate potassium salt.
: Treatment of methyl (2-sulfo-1-methyl)ethyldithio-carbamate potassium ~alt with sodium azide also as described in Example 7 gives 1 (2-sulfo-1-~ethylethyl)- ~.
te~trazole-5-thiol.
7-D-Mandelamidocephalospora~Lc acid and 1-(2-sulfo-1 methylethyl3tetrazole-5-thiol are reac~ed in the presence ; 20 o~ excess sodium bicarbonate as described in Example 7 to give the title compound.
EXAMPLE 20 ~
Reaction o~ the sodium salt of a cephalosporanic ~ :
;~ 25 acid listed below: ~:
7-(2,2,2-trifLuoroethylthioacetamido)~
cephalosporanic acid 7-methylthioace~amidocephalo~poranic acid 7-n-propylthioacetæmidocephalosporanic acid ~`
with 1-(2-sulamoylethyl)tetrazole-5-thiol sodium salt by :
:~:
the procedure of Example 14 gives the 7-subs~ituted-3-~ '~
: - 32 - ;--: .

1~ 78 i~
' - ,'; ~,.
[1-~2-sulfamoylethyl)teJcrazol-5-ylthiomethyl]-3-cephem-4 carboxylic acids listed below~
7- (2, 2, 2-trifluoroethylthioace~amido) -3- ~1- (2- `
sulfamoylethyl~ te~:razol-5 -ylthiomethyl ¦ - 3-cephem-4-carboxylic acid C15H17F3~76S4 ' Na 0 . 5 CH40 Calculated: 30.24% C; 3.11% H; 15.92% N :~
Found: 30.62% C; 3.00% H; 15032% N `~
7-methylth~oacetamido-3~[1-(2-sulfamoylethyl)- ~ ~
tetrazol-5-ylthiomethyl]-3-cephem-4-carbox~lic acid -: -7-n-propylthioacetamido-3-[1-(2-sul~amoylethyl)-~etrazol^5-ylthiomethyl]-3-cephem-4~carboxylic acid. : `
: EXAM~IE 21 :
j_(2-Th ~ ~ etrazol-5-Ylthio- -.

Reaction of 2.18 g. (0.01 mol.) of l-sulfomethyl-tetrazole-S-thiol sodium salt, 5.23 g~ (0.012 mol.) o~ 7 ~2-thienylacetamido)cephalosporanic acld sodium ~alt and .84 g.: (O.OI;mol.) of sodium blcar~onate as described .
2~ in the pro edure of Example 14 ga~re the title compound ;~;
:
as the corresponding disodlum salt~
C16U14N67S4 2 Na 0 75 C2H6 Calculated: 34.81% C; 3.09% H; 13.56% N
F~u~d: 34.41% C; 3.03% H; 13.76% N
7- (2-Thlenylacetamido) -3~ sul~omethyl tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid disodium salt ...
is converted to the title compound as described above.
.; _E 22 Reac~Lon of 5.68 g. (O.ûl mol.) of l-t2-sulfoethyl~-- ~ , .. . . . ., . ... . ,, . ,, ;.... . . .. .. .

~ Z0~8 tetrazole-5-'chiol cyclohexylamine salt and 5 . 23 g . (û . 0125) mol.) of 7-(2-t~ienylace~amido)cephalosporanic acid sodium :~
salt in 80 ml, of water containing sufficlent sodium bicarbonate to ma~ntain the pH at 7.0, as described i~
thc procedure o Example 14, gave the ti~le compound. ~`
EXAMPLE 23 :

To a mixture o~ 97 g. (200 ml., 2,1 mol.3 o~ formic acld, di~tilled from a~hydrous copper sulfate, and 37.5 ml.
(0.4 mol.) of acetic anhydride was added 25.0 g. (0.1 mol.) o~ 7 aminocephalosporanic acid. The mixture was stirred : at ambient ~emperature for 0.5 hour, ~hen evaporated to ; dryness. The res~due was dissolved Ln ethyl acetate and the ethyl acetate solution was filtered and evaporat~d ~o dryness to give a residue which was recrystalliz~d from ether-petroleum ether to gi~re 7-formamidocephalo-sporanlc acid. :.
A mixture of 1.0 g. (3.3 ~nol.~ of 7-formamido-cephalosporanic acid and 0.7 g. ~2.6 mmol.) of l-sulfo-methyltetrazole-5-thiol disodium sal~ in 15 ml. of wa~er wàs stirred a~ 6S 70 for 3 hours while maintaixling the pH at 7.0 by addition o~ sodium bicarbonate and/or hydrochloric acid. The mixture was cooled, acidified `
to pH 1.0 with hydrochloric acid and extrac~ed wi~h ethyl . ~
acetate. The extract was filtered and the filtrate was evaporated to dryness to give a residue which was dis-solved in 30 ml. of methanol. The methanol ~olu~ion was fLltered, 30-40 ml. of isoprop~nol and ethanol were ad~ed, the solution was filtered ~gain and 100 ml. of ether was added. Th~ preeip~tate was collected by filtr tion and drled to give ~he ti~le compound. . ~ ~.
- - ~4 -J::

CloH126N6S3 0.33 C3H8O HCl 2 H2O
Calculated: 27.0~ C; 3.50% H; 16.95% N
Found: 26.96% C; 3.23% H; 16.55~ N
EXAMPLE 24 ; ;;
7-(4-Pyridylthioacetamido)-3-(l-sulfomethyltetrazol-5 ~lthiomethyl)-3-cephem-4~carboxylic acid ; (4-Pyridylthio)acetyl chloride (0.53 g., 2.8 mmol.) was dropwise added to a mixture of 1.0 g. of 7-amino-3- , ~l-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4~car- ;-boxylic acid (mixture of sodium salt, free acid and sodium chloride) and 0.9 g. (9.0 mmol.) of triethylamine in 10 ml. of dry dimethylformamide. The reaction mixture ~ -.. .
was stixred for 1.5 hour at -10, then it was warmed to ;
ambient temperature and stirred for 1 hour.; The mixture was filtered and the filtrate was diluted with 200 ml.
of ether-petroleum ether. The precipitate was collected by filtration and dissolved in warm methanol. The methanol solution was filtered and the fil`trate was con-` centrated to 10 ml. Ethanol (50 ml.) was added and the precipitate was collected by filtration and washed with ether to give the title compound as the sodium salt.
; Additional amounts of the salt were obtained by addition of ether to the filtrate.
C17H16N77S4 Na NaCl C6H15N
; Calaulated: 37.23% C; 4.21% H; 15.10% N ;;
Found: 36.84% C; 4.48% H; 15.43% N
7-(4-Pyridylthioacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid sodium salt is converted to the title compound by procedures described above.

.~
'~" ` ''" ;`~
''' `'':

lO~ZQ~
1 ' Using the procedures deseribed above for the prepara~ion o~ compounds of ~his ~nvention, the following compounds were prepared:
7~ tetrazolylace~am~do)-3-(1 sulfomethyl-~etrazol-5-ylthiomethyl) 3-cephem-4-carboxylic acid 13H12N107S3 2 Na ~.
Calculated: 27.76% C; 2015% H; 24.90% N
; Found: 27.55~/~ C; 3.21% H; 23~00~/o N
7-(3-sydnoneacetamido)-3~ sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acld C14Hl~N809S3 2 Na - 3 H20 4 Calculated: 27.59% C; 3072% H; 16.08% N
Found: 27.59% C; 3.02% H; 15.67% N
:.
7-trifluoromethy1thioacetamido-3-[1-(4-sulfo-but~l)tetrazol-5-ylthi~methyll-3~cephem-4-carboxylic acid ~:
l6~17F3N67S4 2 Na ~7o : Calculated: 29.36% C; 2.~3% H; 12.84% N
: :
-. 20 Found: 29.31% C; 3.33% H; 12.76% N
7-(2-thienylacetam~do)-3-[1-(4-sulfobutyl)tetrazol-5-ylthiomethyl]-3~cephem-4-carboxyl~c acid ~l9H20N607S4 2 Na 2 H20 2 C2H60 Calculated: 34.85% C; 3.69% H; 12.83% N;
19.59%
Found: 34.52% C; 3~88% H; 13.54% N; .
. '19o09% S
7-(2-thienylacetamido 3-[1-~5-sulfopen~yl)tetrazol-5-ylthiomethyl]-3-cephem 4-carboxylic acid C2oH22N6o7s4 ~ Na 2.5 H20 - 1.75 C2H60 ' ' .:

Z~7~3 :
Calculated: 36.33% C; 4.77% H; 11.29% N; 17.23% S
Found: 36.46~,~ C; 4.30% H; 11.63% ~1; L7.30% S

An inje ::table pharmaceutical composition i8 formed 5 by adding ~terile water or ~terile ~aline solutior~
(2 ml . ) to 500 mg . of 7-D-mandelamido-3- [ 1 (2-sulamoyl-ethyl~tetrazol-5-ylthiomethyl] -3-cephem~4 s:arboxylic ac 1d sodium salt.
Pharmaceutlcal composi~ions of the other anti~
10 bacterial compounds disclosea above may be ~ormulated in a si{nilar manner.

, . ~:

, ~, . , ~0 . . ~ ~
', : . '. ' ;;, .
' -`~
-, ~ _ 37 _ ;:

~7Z~
SUPPLEMENT~RY DISCLOSU~E -~
The principal disclosure relates to the preparation of cephalosporins having various acyl substituents at the 7-position and a sulfonic acid or sulfamoyl substituted tetrazolyl thiomethyl group at the 3-position of the cephem nucleus as well as intermediates for the preparation thereof.
The compounds prepared by the invention are found to have antibacterial activity.

Further to the teachings of the principal disclosure ; 10 and within the generic concept an additional species has been found and the ~ollowing example and data is provided. ~`~
Example 27 7-(o-Aminomethylphenylacetamido)-3~ sulfomethyltetrazol-S-lthiometh~ 3-ce~hem-4-carboxvlic acid : ~ `
A solution of o-(N-t~butoxycarbonylaminometh~
phenylacetic acid (5.3 g, 20 mmoll, triethylamine ~2.77 ml) and N-methylmorpholine (4 drops) in tetrahydrofuran (150 ml) was cooled to -10 and treated with i-butyl chloroformate ~; 20 (2.72 y, 20 mmol)~ The xesulting solution was stirred for ~;
10 minutes and then an ice cold solution of 7-ACA (5.44 g, 20 mmol) and ~-riethylamine (2.77 ml) in 1:1 tetrahydrofuran-water (100 ml) was added at a rate so that the temperature remained at -10. The reaction was stirred for 3 hours without cooling, the organic solvents were removed by `
distillation and water (100 ml~ was added. The solution was adjusted to pH 7.5 with triethylamine and extracted with ethyl acetate. The agueous phase was layered with ethyl acetate, adjusted to pH 2.3 with 3N HCl and the phases were separated.

The aqueous phase was re-extracted with fresh ethyl acetate and the combined extracts were dried~ Removal of the solvent in vacuo left an oil; 6.0 g.

~ - SD 37a -~0'~20~8 To a solution of NaHC03 (~.4-9 g) in water (75 ml) was added the above product (3.0 g, 5.8 mmol)~ When solution was complete, l-sulfomethyl~etrazole-5-thiol disodium salt was added. The solution was stirred and heated at 60 while maintaining a pH of 7.0 to 7.2 by adding 5% NaHC03 as needed.
After 4.75 hours the reaction was cooled to room temperature and extracted with ethyl acetate. The aqueous solution was adiusted to pH 7.0 and then applied to a column of "Amberlite ~ ;
XAD~7" resin (2" x 34") which had been pre-washed with . .~, :
methanol followed by wa~er. The product was eluted from the column with water at a flow rate of 10 ml per minute.
The fractions containing the product were freeze dried to yield 1.6 9 o t-BOC protected product.
To an ice cold mixture of trifluoroacetic acid (20 ml~ and d~methoxybenzene (20 ml) was added with stirring l;
the above t-BOC protected product (1.6 g). Cooling was discontinued and the reaction was stirred for 20 minutes.
The trifluoroacetic acia was removed in vacuo and the remaining liquid was stirred while fresh ether was added to precipitate the tri~luoroacetate salt of the title compound~

C21H22F3N7gS3 h 3/4 H20 Calculated: 36.92~ C; 3.47~ H; 14.35~ N
Found: 36.80~ C; 3.82% H; 14~43~ N
The product was collected, washed with ether, st.irred vigorously with acetonitrile (100 ml) for 2 hours and then recollected and washed with fresh acetonitrile to give the -salt of the title compound; 880 mg.
-" ~

' - SD 37b -~4 ' :
` ~ 10'72~
BIOLOGICAL DATA ~
~.
Bacteria MIC (~g/ml~ ln vitro S. aureus HH 127 1.6 S. aureus SKF 23390 1.6 S. villaluz ~200 Strep. ~aecalis HH 34358 25 E. coli SKF 12140 0.Z
E. coli HH 33779 0.4 Kleb. pneumo. SKF 4~200 ~0.1 Kleb. pneumo. SKF 1200 CO.l `~
~al. paratyphi ATCC 12176 ~0.1 Pc mirabilis PM 444 0O4 Pseudo. aerug. HH 63 ~200 Serra. marc. ATCC 13880 6.3 Proteus morgani 179 50 : . :
Entero. aerog. ATCC 13048 0.8 Entero. cloacae HH 31254 0.8 -~
' ' . ` . ,.,' .: ~, ' ~ED50 ~
. .
E. coli ~KF 12140) 0.26 mg/kg s.c.
Kleb. pneumo.(5KF 4200) 0.28~ mg/kg s.c.

BLOOD LEVELS
Tl/2 = 80 minutes \ Tl/2 = 78 minutes \
Serum binding = 46% ~ mouse ~squirrel -~ /monkey p D 108 @ 30 minutes / P = 116 @ 15 minutes , `

- SD 37c -~ ~, ~,.
: ' ~

Claims (34)

The embodirnents 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:

in which:
each individual R1 is hydrogen or lower alkyl;
n is one to ten;
R2 is hydroxy, amino, lower alkylamino or di(lower) alkylamino; and R3 is an acyl group selected from the group consisting of , and where:
X is thienyl; phenyl or phenyl monosubstituted with hydroxy;
A is NH2 or OH, Y is thienyl, tetrazolyl, cyano, sydnone or aminomethylphenyl;
Z is methyl, trifluoromethyl, trifluoroethyl, cyanomethyl or pyxidyl; and m is zero, or a non-toxic pharmaceutically acceptable salt thereof, comprising reacting a compound of the formula:

where R3 is defined as above or a salt thereof, suitably protected as necessary, with a compound of the formula:

where R1, R2 and n are defined as above or a salt thereof, or acylating a compound of the formula:

where R1, R2 and n are defined as above and R4 is hydrogen or a protecting ester group, with an appropriate acylating agent of the formula R3-OH or an activated derivative thereof where R3 is defined as above suitably protected as necessary followed by acidification when necessary, removal of the protective group(s) when necessary and optionally converting the product to a non-toxic pharmaceutically acceptable salt.

2. A process as claimed in claim 1 in which R1 is hydrogen and n is one to five.

3. A process as claimed in claim 2 in which R3 is , X is phenyl or hydroxyphenyl, A is NH2 or OH and R2 is hydroxy or amino.

4. A process as claimed in claim 2 in which R3 is , X is thienyl and R2 is hydroxy or amino.

5. A process as claimed in claim 2 in which R3 is , Z is methyl, trifluoromethyl or trifluoro-ethyl and R2 is hydroxy or amino.

6. A process for preparing 7-mandelamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid comprising reacting 7-mandelamidocephalosporanic acid with 1-sulfomethyltetrazole-5-thiol sodium salt and then acidifying.

7. A process for preparing 7-mandelamido-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting 7-mandelamidocephalosporanic acid sodium salt with 1-(2-sulfoethyl)tetrazole-5-thiol and then acidifying.

8. A process for preparing 7-mandelamido-3-[1-(5-sulfopentyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting 7-mandelamidocephalosporanic acid sodium salt with 1-(5-sulfopentyl)tetrazole-5-thiol disodium salt and then acidifying.

9. A process for preparing 7-mandelamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-car-boxylic acid comprising acylating 7-amino-3-[1-(2-sulfamoyl-ethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid with O-dichloroacetylmandeloyl chloride.

10. A process for pre-paring 7-(2-thienylacetamido)-3- (1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid oomprising reacting 7-(2-thienylacetamido)cephalosporanic acid sodium salt with 1-sulfomethyltetrazole-5-thiol sodium salt and then acidifying.

11. A process for pre-paring 7-(2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxvlic acid comprising reacting 7-(2-thienylacetamido)cephalosporanic acid sodium salt with 1-(2-sulfoethyl)tetrazole-5-thiol cyclohexylamine salt and then acidifying.

12. A process for preparing 7-(2-thienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid comprising reacting 7-(2-thienylacetamido)cephalosporanic acid sodium salt with 1-(2-sulfamoylethyl)tetrazole-5-thiol sodium salt and then acidifying.

13. A process for preparing 7-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid comprising reacting 7-trifluoromethylthioacetamidocephalosporanic acid sodium salt with 1-sulfomethyltetrazole-5-thiol sodium salt and then acidifying.

14. A process for preparing 7-trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid compri-sing reacting 7-trifluoromethylthioacetamidocephalosporanic acid sodium salt with 1-(2-sulfamoylethyl)tetrazole-5-thiol and then acidifying.

15. A compound of the formula:

in which:
each individual R1 is hydrogen or lower alkyl;
n is one to ten;
R2 is hydroxy, amino, lower alkylamino or di(lower)alkylamino; and R3 is an acyl group selected from the group consisting of , and where:
X is thienyl; phenyl or phenyl monosubstituted with hydroxy, A is NH2 or OH, Y is thienyl, tetrazolyl, cyano, sydnone or aminomethylpheny;
Z is methyl, trifluoromethyl, trifluoroethyl, cyanomethyl or pyridyl; and m is zero, or a non-toxic pharmaceutically acceptable salt thereof, when prepared by the process of claim 1 or its obvious chemical equivalent.

16. A compound of claim 15 in which R1 is hydrogen and n is one to five, when prepared by the process of claim 2 or its obvious chemical equivalent.

17. A compound of claim 15 in which R1 is hydrogen, n is one to five, R3 is , X is phenyl or hydroxyphenyl, A is NH2 or OH and R2 is hydrogen or amino, when prepared by the process of claim 3 or its obvious chemical-equivalent.

18. A compound of claim 15 in which R1 is hydrogen, n is one to five, R3 is , Y is thienyl and R2 is hydroxy or amino, when prepared by the process of claim 4 or by its obvious chemical equivalent.

19. A compound of claim 15 in which R1 is hydrogen, n is one to five, R3 is , Z is methyl, trifluoro-methyl or trifluoroethyl and R2 is hydroxy or amino, when pre-pared by the process of claim 5 or its obvious chemical equi-valent.

20. The compound 7-mandelamido-3-(1-sulfomethyl-tetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid, when prepared by the process of claim 6 or its obvious chemical equivalent.

21. The compound 7-mandelamido-3-[1-(2-sulfoethyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid, when prepared by the process of claim 7 or its obvious chemical equivalent.

22. The compound 7-mandelamido-3-[1-(5-sulfopentyl)-tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid, when prepared by the process of claim 8 or its obvious chemical equivalent.

23. The compound 7-mandelamido-3-[1-(2-sulfamoyl-ethyl)tetrazol-5-ylthiomethyll-3-cephem-4-carboxylic acid, when prepared by the process of claim 9 or its obvious chemical equivalent.

24. The compound 7-(2-thienylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4 carboxylic acid, when prepared by the process of claim 10 or its obvious chemical equivalent,

25. The compound 7 (2-thienylacetamido)-3-[1-(2-sulfoethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid, when prepared by the process of claim 11 or its obvious chemical equivalent.

26. The compound 7-(2-thienylacetamido)-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthiomethyl]-3-cephem-4-carboxylic acid, when prepared by the process of claim 12 or its obvious chemical equivalent.

27. The compound 7-trifluoromethylthioacetamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid) when prepared by the process of claim 13 or its obvious chemical equivalent.

28. The eampound 7-trifluoromethylthioacetamido-3-[1-(2-sulfamoylethyl)tetrazol-5-ylthlomethyl]-3-cephem-4-carboxylic acid, when prepared by the process of claim 14 or its obvious chemical equivalent.

29. The process as claimed in claim 3 for preparing 7-D-mandelamino-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid comprising reacting 7-D-mendelamidocephalosporanic acid with 1-sulfomethyltetrozole-5-thiol sodium salt.

30. The process as claimed in claim 6 for preparing 7-mandelamido-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid disodium salt comprising treating the 7-mandelamido-3-(1-sulfomethyltetxazol-5-ylthiomethyl)-3-cephem-4-carboxlic acid with sodium methoxide.

31. The compound 7-D-mandelamino-3-(1-sulfomethyletetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 29 or its obvious chemical equivalent.

32. The compound 7-mandelamide-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid disodium salt when prepared by the process of claim 30 or its obvious chemical equivalant.

Claims Supported by Supplementary Disclosure

33. A process as claimed in claim 1 for preparing 7-(o-aminomethylphenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-(o-N-t-butoxycarbonylaminomethylphenylacetamido)-cephalosporanic acid with 1-sulfomethyltetrazol-5-thiol disodium salt and then removing the protecting group with trifluoroacetic acid.

34. 7-(o-aminomethylphenylacetamido)-3-(1-sulfomethyltetrazol-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 33 of its obvious chemical equivalent.