Note: Descriptions are shown in the official language in which they were submitted.
~ 3~
The present invention provides a process for the
displacement of the acetoxy group of a cephalosporanic acid
by a sulfur nucleophile, in an organic solvent and under
essentially anhydrous conditions.
The displacement of the acetoxy group of a cepha-
losporin by a sulfur nucleophile is a known reaction (U.S.
Patent 3,278,531). This patent and other publications (J.
D. Cocker, J. Chem. Soc~, 1965, 5015) teach that the re-
action occurs only in an aqueous medium. Practical dis- --
placements have utilized a salt of the cephalosporanic acid
in water along with the sulfur nucleophile or its salt at pH
S-8. The combination of aqueous medium, elevated temper-
ature (35-70C.), and near neutral to basic pH are generally
destructive of much of the cephalosporin nucleus, and
products prepared in this manner often require extensive
purification. Attempts to conduct the displacement on
cephalosporanic acids in water at lower pH (p~ 2-3) lead to
substantial lactone formation, a side-reaction that dramatically
lowers the yield of desired product.
It has now been discovered that the displacement
of the acetoxy group (as well as other 3-acyloxy groups) of
cephalosporanic acids by sulfur nucleophiles can be achieved
in organic solvents under essentially anhydrous conditions.
Reactions under these conditions are not complicated by -
lactone formation; yields are generally higher; and products
are more easily isolated. Some of the products spontaneously - -
precipitate from the reaction mixture. The present process
is believed to be general in nature, and applicable to
~ essentially any sulfur nucleophile and any cephalosporanic
acid.
X~ 02A
.
- - . . . . . .
: ~ .. :
3~
The object of thls invention i8 to provide an
- improved pr~cess for preparing 3-(thiomethyl)cephalosporins.
The present invention is a novel process for
preparing a cephalosporin compound of the formula
R~
R~ 7 t
CH SR~ 3
OOH
wherein R13 is - :
1 0 /R4
,~R4
~4
+ 4 .
-~-o~
. ' '
., ~ , .
-C -C a I ky I,
,,, - ..
~(R5)o 1 ~r ~; or - ~ .
,:
:
: ~ ,. '
l is hydrogen or methoxy;
' ' ' ' '
'~'',
X-4902A 3
., . ' .~
..
,--................................................... I .
.. . . . . . . . . .
~ . .
: , ' .: ~ . ' ', . . ~ ' ;
.,
~31~a1
R2 is phthalimido, succinimido, a radical of the
formula ~ --fH--f wherein L is hydrogen or nitroso,
L~
CH3 CH3
or a radical of the formula
.
R -C-NH-;
R is
(1) hydrogen, -
: (2) Cl-C6 alkyl,
(3) -CH2 - (Cl-C3 chloroalkyl),
(4) -CH~ - (Cl-C3 fluoroalkyl),
(5) Cl-C4 cyanoalkyl,
(6) Cl-C4 hydroxyalkyl, :
(7) p-nitrobenzyloxy, . . .
~ (8) tert-butoxy, -
:~ : (9) (2,2,2-trichloroethoxy), -
(10) a protected 4-amino-4-carboxybutyl :
radical of the formula
o ,
R12O C CH (CH ) CH
~: A
wherein Al is a protected amino group : :-
: and R12 is hydrogen or Cl-C4 alkyl; :
: (11) 4-oxo-4-carboxybutyl;
:; (12) 3-carboxypropyl;
(13) a radical of the formula ~ :~
a . .
X-4902A
- . .
.
~0~33~
in which each of a and al i5 independently
hydrogen, Cl-C4 alkyl, Cl-C4 alkoxy,
halogen, hydroxy, or AlCH2- wherein
is, as above, pro~ected amino; Z is o
or S; and m is 0 or 1;
(14) a radical of the formula
P-CH-
Q
wherein P is
(a) 2-thienyl,
(b) 3-thienyl, or
(c) a phenyl group of the formula
~< _ / : ' ,
1 o__a ~ .
in which a and al are as defined above;
and
wherein Q is :
(a) hydroxy,
(b) formyloxy, -: ;
(c) acetoxy, . ~ .
(d) carboxy of the formula -C-O-A2 wherein
A2 is diphenylmethyl, p-nitrobenzyl,
~; benzyl, 2,2,2-trichloroethyl, tert-butyl,
or p-methoxybenzyl; ~ ~ :
(ej (alkali metal oxy~ulfonyl), - .: :
() a protected amino group, .
~ ~ ' , ' , ' ,'
X-4902A 5
3~
(g) an acylated amino group of the formula
O
-NH-C-T wherein T represents amino,
NH R O
" ' " 8
-NH-C-NH2, -N--C-R ,
R7 o
-N -C-CH=CHR9 ~f
\N ~
R10 . : .
1 0 '~"" ' ' ' ' '
/~=~ ~--tOH) 2 or 3; ~r
R1o . ., ' '
, ~
. '
wherein R7 is hydrogen or Cl-C3 alkyl; :
R8 is phenyl, halophenyl, furyl, mono- ~ :
methylamino, dimethylamino, monoethyl- :.
amino, diethylamino, methylethylamino, ~:
propylamino, dipropylamino, diisopropyl-
amino~ phenylamino, or diphenylamino; R9
is hydrogen, Cl-C~ alkyl, or phenyl; Rl0
is hydrogen, Cl-C3 alkyl, or methyl-
sulfonyl; and Rll is ethyl.ene, tri-
methylene, or vinylene;
X-4902A -6
, . . ~.. ~ . . . . . .
-` ~V83~43.
(15) a radical of the formula
o
N_oH~ -OCH3, -oCcH3
P --C--C-- .
O : ' '
wherein P' is P as defined above, pro-
tected 2-amino-4-thiazolyl, or 2-furyl;
.. . ~ .
(16) a radical of the formula V-S(O)n-CH2-
wherein V represents -CF3 or -CH2-X :~
wherein X represents hydrogen, methyl, .
CF3, CN, or N3, and n represents 0, l,
or 2; or
(17) a radical of the formula Y-C~2- in which
is
2~thienyl, -
3-thienyl, ~ :
2-furyl, . -:
2-oxazolyl,
: ~ 2-thiazolyl, :~
l-tetrazolyl, . -.
l-benzotriazolyl,
: . 2-oxazolylthio,
. . .
2-thiazolylthio,
1,2,3-triazol~5-ylthio,
1,3,4-triazol-2-ylthio,
1,3,4-thiadiazol-2-ylthio,
protected 5-amino-1,3,4-thia-
diazol-2-ylthio,
. ~. .
; 5-methyl-1,3,~-thiadiazol-2-
. ylthio,
1,2,4-thiadiazol-5-ylthio,
.~ . .
X-4902A ~7~ . ~ ~
. ', ~ '.
': .
. . . ~ - . . :, . : , . .
... .. :, . -.......... :. . .. . :
~o~
3-methyl-1,2,4-thiadiazol 5-
ylthio,
1,2,5-thiadiazol-3-ylthio, .
1,3,4-oxadiazol-2-ylthio, ~
5-methyl-1,3,4-oxadiazol-2- : -
ylthio,
l-methyl-5-tetrazolylthio,
pyridylthio, ~ .
4-cyano-1,2,3-triazol-1-yl,
3-cyano-1,2,4 triazol-1-yl, or
protected 2-amino-4-thiazolyl, ~ .
each R is independently
hydrogen, .
Cl-C4 alkyl, ~ :~
C2-C3 aIkenyl,
cyclohexyl, or ~::
phenyl; .
each R5 is independently Cl-C4 alkyl, Cl-C
alkoxy, halo, hydroxy, nitro, cyano, methane- :
sulfonamido, or trifluoromethyl; and
R6 is
~ N_
a unit:which with the -C
comprises an unsubstituted or substituted,
five or six-membered, heteroaromatic ring
having a total of from 1 to 4 hetero atoms
selected from the following combinations:
1 nitrogen and 0 or 1 oxygen or sulfur,
2 nitrogens and 0 or 1 oxygen or sulfur,
3 nitrogens, and 0 or 1 oxygen, or
4 nitrogens, ..
all other ring atoms being carbon; or
':'
: X-4~02~ -8-
'. ~,
', - , :, :,, . ,' : :
. . . .
33~
a unit which with the -C
comprises 2-benzimidazolyl, 2-benzothiazolyl,
2-benzoxazolyl, or a radical of the formula ~ -
N=N
$~
which process comprises reacting a compound of the formula
R2
~ CH~OCOR II
OOH - .
wherein Rl and R2 are as defined above, and
R lS
cl-C3~alk~
C4-C6 cycloalkyl,
amlno, - .
mono- or di(Cl-C3 alkyl)amino, :~
-C = CH~ ~-OH
OCH
3 ~:
C = CH-~ ---OH
OCH ~ , or . .
. .
~: -C = CH--~ ~/O--OSO3H;
OCH~
;~
: X- 4902A _g_
: ,' , ~:
3~
with a sulfur nucleophile o the formula
R13_s_R14 III
wherein R13 is as defined above, and R14 is hydrogen, or
when and only when R13 is a methyleneaminium group, R14
combines with R13 to form a thiourea; ~
in an organic solvent under essentially anhydrous con- ~ -
ditions.
The products of the present process exhibit anti-
bacterial activity; in addition, many of them serve as
intermediates to yet other cephalosporin products which also
exhibit antibacterial activity, see, e.g. U.S. 3,932,393.
In the foregoing dPfinitions, the various "alkyl"
terms refer to both straight and branched chain alkyl
groups. In the definition of R3 as "Cl-C6 alkyl," alkyl
refers to groups such as methyl, ethyl, propyl, isopropyl,
butyl, sec-butyl, amyl, isoamyl, hexyl, 2,3-dimethylbutyl,
and the like. The terms "-CH2-(Cl-C3 chloroalkyl)" and ~; -
"-CH2-(Cl-C3 fluoroalkyl)" refer to such groups as chloro-
ethyl, fluoroethyl, 2-chloroethyl, 2-chloropropyl, 3-fluoro-
propyl and 4-chlorobutyl. The term "Cl-C4 cyanoalkyl"
refers to such groups as cyanomethyl, 2-cyanoethyl, 3-
cyanopropyl and 2-cyanopropyl. The term "Cl-C4 hydroxy
, .
alkyl" refers to such groups as hydroxymethyl, 2-hydroxy-
ethyl, 3-hydroxypropyl and 2-hydroxypropyl.
In both the 3-(acyloxymethyl)aephalosporin of
Formula II and the sulfur nucleophile, amino groups are
desirably protected. The practice of protecting amino
.
groups is well known. See Protective Groups In Organic
Chemistry, edited J. T. W. McOmie (Plenum Press, L,ondon and
. .
X-4902A -10-
~ .. .
'' ': ~ '
.
. .. ' . : : . . . .: :
83~
New York, 1973). ~n general, protection o an amino group
implies removability of the protection group; but that is
not necessary as, for example, in a 7-acylamido radical of
the ~ormula
O o
12 " ~
R O-C-cH-(cH2)2-cH2-c-NH
which is subsequently to be cleaved off of the cephalosporin.
Suitable protecting groups for this radical include C2-C4
alkanoyl, C3-C4 chloro- or fluoroalkanoyl, benzoyl, and ~-
substituted benzoyl. The term "C2-C4 alkanoyl" refers to
acetyl, propionyl, butyryl, and ~he like. The term "C3-C4
chloro- or fluoroalkanoyl" refers to 3-chloropropionyl,
3-fluoropropionyl, and the like. The term "substituted
benzoyl" includes halo substituted benzoyl groups such as 4-
chlorobenzoyl, 4-bromobenzoyl, and 2,4-dichloroben~oyl.
Such groups can also be employed for the protection of amino
groups at other locations in the 3-(acyloxymethyl)ceph- -
alosporin, and in the sulfur nucleophile. However, when it `
is desired to regain a free amino, the protecting group `~
should be one which is readily removable. See McOmie,
supra. Suitable protecting groups which are readily
removable are benzyloxycarbonyl, p-nitroben~yloxycarbonyl,
2,2,2-trichloroethoxycarbonyl, tert-butoxycarbonyl, p~
methoxybenzyloxycarbonyl, and diphenylmethoxycarbonyl. The
amino group of the formula
O O :
R O C-C,Hl-(CH2)2-CH2-C-NH- ~ ;
can also be protected as described in Belgian patent 771,694.
-4902A -11-
; ~, ~'' .
:: . ~
. ',. . `-' ' '' ', ' ; ' ' , " . ` , ' ~:, .:
.. .: .. , , . , . , . : .
3~
As used herein, the term "halogen" and the term
"halo" each refers to fluoro, chloro, bromo, or lodo. The
term "Cl-C4 alkoxy" refers to methoxy, ethoxy, isopropoxy,
n-butoxy, and the like. The ~erm "alkali metal" preferably
designates sodium, potassium, and lithium.
The following are illustrative of the group
R3-C-NH- wherein R3 is any of moieties (1) throu~h (12):
formamido, acetamido, propionamido, butyramido, a-methyl-
propionamido, valeramido, a-methylbutyramido, trimethylacet-
~0
amido, hexanamido, heptanamido, 3-chloropropionamido,
3-chlorobutyramido, 4-fluorobutyramido, 5-chlorovaleramido,
cyanoacetamido, 2-cyanopropionamido, 3-cyanopropionamido,
4-cyanobutyramido, hydroxyacetamido, 2-hydroxypropionamido,
3-hydroxybutyramido, p-nitrobenzyloxycarbonylamino, tert-
butoxycarbonylamino, (2,2,2-trichloroethoxy)carbonylamino,
5-(~,5-dichlorobenzamido)-5-carboxyvaleramido, 5-acetamido-
5-carboxyvaleramido, 5-carbomethoxy-5-(2,4-dichlorobenz-
amido)valeramido, 5-carbo-n-butoxy-5-(2,4-dichlorobenz-
amido~valeramido, 5-carboxy-5-(2,4-dichlorobenzamido)-
valeramido, 5-(p-chlorobenzamido)-5-carboxyvaleramido,
5-propionamido-5-carboxyvaleramido, 5-(3-chloropropion-
amido)-5-carboxyvaleramido, 5-benzamido-5-carboxyvaleramido,
5-oxo~5-carboxyvaleramido and ~-carbox~butyramido.
The following are illustrative of the groups
R3-C-NH- in the above definition in which R3 is
;~ a\ ~_
~ ~ ~-(Z)~-CH~-
a~
X-4902A -12-
'i"' ' ~ .
: .
3~4~
and in which m is O: phenylacetamido, 2-(p-methylphenyl)-
acetamido, 2-(m-ethylphenyl)acetamido, 2-tp-isopropyl-
phenyl)acetamido, 2-to-methylphenyl)acetamido, 2-(p-chloro-
phenyl)acetamido, 2-tp-bromophenyl)acetamido, 2-(2,4-
dichlorophenyl)acetamido, 2-(m-bromophenyl)acetamido,
2-(p-fluorophenyl)acetamido, 2-(o-fluorophenyl)acetamido,
2-t3,4-dihydroxyphenyl)acetamido, 2-(p-hydroxyphenyl)acet-
amido, 2-(m-hydroxyphenyl)acetamido, 2-(2,6-dimethoxyphenyl)-
acetamido, 2-(m-methoxyphenyl)acetamido, 2-(p-isopropoxy-
phenyl)acetamido, 2-(m-ethoxyphenyl)acetamido, 2-tp-methoxy-
phenyl)acetamido, 2-t3,4-dimethoxyphenyl)acetamido, 2-(p-
tert-butoxyphenyl)acetamido, 2-(p-(acetamidomethyl)phenyl)-
acetamido, 2-tp-ttert-butoxycarbonylaminomethyl)phenyl)- :
acetamido, 2-to-ttert-butoxycarbonylaminomethyl)phenyl)- .
acetamido, 2-tm-butoxyphenyl)acetamido and 2-(3-chloro-4- ~.
methylphenyl)acetamido. When, in the above formula, m = 1
and Z represents -0-, illustrative groups inelude the fol-
lowing: phenoxyacetamido, 2-(p-methylphenoxy)aeetamido,
2-(m-ethylphenoxy)acetamido, 2-(p-isopropylphenoxy)acet-
20 amido, 2-(o-methylphenoxy)acetamido, 2-(p-chlorophenoxy)- ::
acetamido, 2-(p-bromophenoxy)acetamido, 2-(2,4-dichloro-
phenoxy)acetamido, 2-(m~bromophenoxy)aeetamido, 2-(p- :~ ;
fluorophenoxy)aeetamido, 2-(o-fluorophenoxy)aeetamido,
2-t2,6-dimethoxyphenoxy)aeetamido, 2-tm-ethoxyphenoxy)aeet-
amido, 2-(p-methoxyphenoxy)acetamido, 2-t3,4-dimethoxyphenoxy)-
acetamido, 2-tp-tert-butoxyphenoxyjaeetamido, 2-to-butoxy-
phenoxy)acetamido, 2-t3-chloro-4-methylphenoxy)acetamido,
2-(3-hydroxy-4-methylphenoxy)acetamido, 2-to-ehlorophenoxy)-
aeetamido, 2-t3-hydroxy-4-methylphenoxy)acetamido, 2-to-
X-4902A -13-
', ...
, . .
.... .
~ .
. - . .
33~
chlorophenoxy)acetamido, 2-(p-isopropo~yphenoxy)acetamido,
2-(o-acetamidomethyl)phenoxy)acetamido and 2-(p-(tert-
butoxycarbonylaminomethyl)phenoxy)acetamido. When in the
foregoing formula, m = l and Z represents -S-, illustrative
groups include the following: 2-(phenylthio)acetamido,
2-(p-methylphenylthio)acetamido, 2-(m-ethylphenylthio)acet-
amido, 2-(p-isopropylphenylthio)acetamido, 2-(o-methyl-
phenylthio)acetamido, 2-(p-chlorophenylthio)acetamido,
2-(p-bromophenylthio)acetamido, 2-(2,4-dichlorophenylthio)-
acetamido, 2-(m-bromophenylthio)acetamido, 2-(p-fluoro-
phenylthio)acetamido, 2-(o-fluorophenylthio)acetamido,
2-(3,4-dihydroxyphenylthio)acetamido, 2-(p-hydroxyphenyl-
thio)acet~nido, 2-~m-hydroxyphenylthio)acetamido, 2-(2,6-
dimethoxyphenylthio)acetamido, 2-(m-ethoxyphenylthio)acet-
amido, 2-(p-methoxyphenylthio)acetamido, 2-(3,4-dimethyl-
phenylthio)acetamido, 2-(p-tert-butoxyphenylthio)acetamido,
2~(m-butoxyphenylthio)acetamido, 2-(3-chloro-4-methyl-
phenylthio)acetamido, 2-(3,4-dimethylphenylthio)acetamido,
2-(3,4-dichlorophenylthio)acetamido, 2-(2,5-dichlorophenyl- -~
thio)acetamido, 2-(3-fluoro-4-chlorophenylthio)acetamido,
2-(3-chloro-4-fluorophenylthio)acetamido, 2-(2,6-difluoro
phenylthio)acetamido, 2-(m-fluorophenylthio)acetamido,
2-(o-(acetamidomethyl)phenylthio)acetamido and 2-(p-(tert- ~ -
butoxycarbonylaminomethyl)phenylthio)acetamido.
When R3 represents a group of the formula P-CH-
Q
O
illustrative groups having the overall formula R3-C-NH-
include the mandelamido group, and the O-formyl and O-acetyl
.. . .
X-4902A -14-
, ' 1 .
,
~C~83~4~
derivatives thereof; the 2-carboxy-2-phenylacetamido group;
the 2-sulfo-2-phenylacetamido group; the protected 2-
amino-2 phenylacetamido group; in which the protected amino
group is, for example, carbonylamino, tert-butoxycarbonyl-
amino, trichloroethoxycarbonylamino, or p-nitrobenzyloxy- ~-~
carbonylamino; and the 2-(acylated amino)~2-phenylacetamido
group. Also included are those 2-substituted 2-thienyl- : :
acetamido and 3-thienylacetamido groups in which the phenyl
group is replaced by a 2-thienyl or a 3-thienyl ring.
Illustrative of the foregoing acetamido groups are
mandelamido, p-methylmandelamido, p-hydroxymandelamido,
m-hydroxymandelamido, p-methoxymandelamido, m-bromomandel-
amido, p-chloromandelamido, 3-methyl-4-fluoromandelamido,
o-fluoromandelamido, p-~luoromandelamido, p-isopropyl-
mandelamido, 3,4-dimethyl-O-formylmandelamido, p-chloro-O- :
: formylmandelamido, m-isopropoxy-o-formylmandelamido, m- ~ ~
- bromo-O-formylmandelamido, O-formylmandelamido, 3,4- :~ -
dlmethoxy-O-formylmandelamido, O-acetylmandelamido, p-
hydroxy-O-acetylmandelamido, p-(acetamidomethyl)mandelamido, :
20 p-hydroxy-O-formylmandelamido, 2-hydroxy-2-(2-thienyl)- ~ :
acetamido, 2-hydroxy-2-(3-thienyl)acetamido, 2-formyloxy-
2-(2-thienyl)acetamido, 2-acetoxy-2-(2-thienyl)acetamido, 2-
formyloxy-2-(3-thienyl)acetamido, 2-acetoxy-2-(3-thienyl)- :
acetamido, 2-(tert-butoxycarbonyl)-2-phenylacetamido, 2- -
(2,2,2-trichloroethoxycarbonyl)-2-(p-methylphenyl)acetamido, :
2-(benzyloxycarbonyl)-2-(p-hydroxyphenyl)acetamido, 2- .
(tert-butoxycarbonyl)-2-(p-hydroxyphenyl)acetamido, 2-(p- :
: nitrobenzyloxycarbonyl)-2-(m-hydroxyphenyl)acetamido,
.. .. .. . .
:.
X-4902A -15-
. .
.. . - . . . .. .. . . . . .
. ~: . :
: ' '' '' ' ' . ... ''':', ' ''"'' "' '.' , .
... . ... . ..
3~L4~
2-(p-methoxybenzyloxycarbonyl)-2-(p-me~hoxyphenyl)acetamido,
2-(diphenylmethoxycarbonyl)-2-(m-bromophenyl)acetamido,
2-(tert-butoxycarbonyl)-2-(p-chlorophenyl)acetamido, 2-
(2,2,2-trichloroethoxycarbonyl)-2-(3-methyl-4-fluorophenyl)-
acetamido, 2-(benzyloxycarbonyl)-2-(o-fluorophenyl)acetamido,
2-(p-nitrobenzyloxycarbonyl)-2-(p-fluorophenyl)acetamido,
2-(p-methoxybenzyloxycarbonyl)-2-(p~isopropylphenyl)acetamido,
2-(tert-butoxycarbonyl)-2-(3,4-dimethylphenyl)acetamido, ~.
2-(tert-butoxycarbonyl) 2-(m-isopropoxyphenyl)acetamido, :
2-(diphenylmethoxycarbonyl)-2-(3,4-dimethoxyphenyl)acetamido,
2-(tert-butoxycarbonyl)-2-(p-(2,5-dichlorobenzamidomethyl)-
phenyl)acetamido, 2-(tert-butoxycarbonyl)-2-(2-thienyl)- :
acetamido, 2-(tert-butoxycarbonyl)-2-(3-thienyl)acetamido,
2-(sodiooxysulfonyl)-2-phenylacetamido, 2-(sodiooxysulfonyl)-
2-(p-methylphenyl)acetamido, 2-(potassiooxysulfonyl)-2-(p-
hydroxyphenyl)acetamido, 2-(lithiooxysulfonyl)-2-(m-hydroxy-
phenyl)acetamido, 2-(sodiooxysulfonyl) 2-(p-methoxyphenyl)- .
acetamido, 2-(sodiooxysulfonyl)-2-(m-bromophenyl)acetamido,
2-(sodiooxysulfonyl)-2-(p-chlorophenyl)acetamido, 2-(sodiooxy-
sulfonyl)-2-(3-methyl-4-fluorophenyl)acetamido, 2-(sodiooxy-
sulfonyl)-2-(o-fluorophenyl)acetamido, 2-(sodiooxysulfonyl)-
2-(p-fluorophenyl)acetamido, 2-(sodiooxysulfonyl)-2-(p-
: acetamidomethylphenyl)acetamido, 2-(sodiooxysulfonyl)-2-
(p-isopropylphenyl)acetamido, 2-(sodiooxysulfonyl)-2-(3,4-
dimethylphenyl)acetamido, 2-(sodiooxysulfonyl)~2~(m-iso-
propoxyphenyl)acetamido, 2-(sodiooxysulfonyl)-2-(3,4-
dimethoxyphenyl)acetamido, 2-(sodiooxysulfonyl)-2-(2-
thienyl)acetamido, 2-(potassiooxysu~fonyl)-2-(3-thienyl)~
acetamido, 2-(p-nitrobenzyloxycarbonylamino)-2-phenylacet-
X-4902A -16-
~ . . .
.. ... . .
.,,,:", ~ :. .,-., , . .- , . , ' '' :
~ 513~
amido, 2-(tert-butoxycarbonylamino)-2-(2-thienyl)acetamido,
2-(benzyloxycarbonylamino)-2-(m-hydroxyphenyl)acetamido,
2-(tert-butoxycarbonylamino)-2-(p-hydroxyphenyl)acetamido,
2-ureido-2-phenylacetamido, 2-ureido-2-(2-thienyl)acet-
amido, 2-(3-guanyl-1-ureido)-2-phenylacetamido, 2-(3-
methylaminocarbonyl-l-ureido)-2-phenylacetamido, 2-(3-
dimethylaminocarbonyl-3-methyl-1-ureido)-2-phenylacetamido,
2-[N-(imidazolidine-2-one-l-ylcarbonyl)amino]-2-phenyl-
acetamido, 2-[N-(3-methylimidazolidine-2-one-1-ylcarbonyl)-
amino]-2-phenylacetamido; 2-[N-(hexahydropyrimidine-2-
one-l-ylcarbonyl)amino]-2-phenylacetamido; 2-[N-(3-methyl- ~
hexahydropyrimidine-2-one-1-ylcarbonyl)amino]-2-phenylacet- ~ :
amido; 2-[N-(3-methanesulfonylhexahydropyrimidine-2-one-
l-ylcarbonyl)amino]-2-phenylacetamido; 2-(3-di-n-propyl- :
aminocarbonyl-l-ureido)-2-phenylacetamido, 2-((4-ethyl-2,3-
dioxo-l-piperazinyl)carbonylamino)-2-phenylacetamido, 2-
(3-(methylsulfonyl)-2-oxo-l-imidazolidinyl)carbonylamino)-
2-phenylacetamido, 2-(2-oxo-4-imidazolin-1-yl)carbonyl-
amino)-2-phenylacetamido, 2-((4-oxo-4H-thiopyran-3-yl)-
20 carbonylamino3-2-phenylacetamido, 2-(3-methyl-3-(methyl- .
: carbamoyl)ureido)-2-phenylacetamido, 2-(3-methyl-3-(cinnamoyl-
ureido))-2-phenylacetamido, 2-(3-me~hyl-3-(acryloylureido))-
2-phenylacetamido, 2-((4-ethyl-2,3-dioxo-l-piperazinyl)-
carbonylamino)-2-thienylacetamido, 2-((q-ethyl-2,3-dioxo-
l-piperazinyl)carbonylamino)-2-(p-hydroxyphenyl)acetamido,
2-((4-ethyl-2,3-dioxo-l-piperazinyl)carbonylamino)-2-phenyl-
acetamido, 2-((2-oxoimidazolidin-l-yl)carbonylamino)-3-
thienylacetamido and 2-((2-oxo-3-(methylsulfonyl)-4-imidazolin-
l-yl)carbonylamino)-2-(p-hydroxyphenyl)acetamido.
:
X-4902A -17-
. . - - - .. . .
: , . : . : .: .
: . . . . .
.. . , ~ :, , . .
.. . . . . . . .
.
~L083~
The following are illustrative of the group
R3-C-NH- wherein R3 is moiety (15): 2-(hydroxyimino)-
2-phenylacetamido, 2-(methoxyimino)-2-phenylacetamido,
2-(acetoxyimino)-2-phenylacetamido, 2-(hydroxyimino)-2-
(2-thienyl)acetamido, 2-(hydroxyimino)-2-(2-uryl)acetamido,
2-(methoxyimino)-2-(3-thienyl)acetamido, 2-(methoxyimino)-
2-(2-~uryl)acetamido, 2-(methoxyimino)-2-(p-hydroxyphenyl)-
acetamido, 2-(hydroxyimino)-2-(2-(2,2,2-trichloroethoxy-
carbonylamino)-4-thiazolyl)acetamido, (tautomeric with -
2-(hydroxyimino)-2-(2-(2,2,2-trichloroethoxycarbonylimino)-
4-thiazolin-4-yl)acetamido), 2-(methoxyimino)-2-(2-(p-
nitrobenzyloxycarbonylamino)-4-thiazolyl)acetamido (tauto-
meric with 2-(methoxyimino)-2-(2-(p-nitrobenzyloxycarbonyl-
imino)-4-thiazolin-4-yl)acetamido) and 2-(methoxyimino)-2- : :
(4-chlorophenyl~acetamido.
The following are illustrative of the group ~. -.
O
R -C-NH- wherein R3 is moiety (16): 2-(trifluoromethyl-
thio)acetamido, 2-(methylsulfonyl)acetamido, 2-(cyanomethyl-
thio)acetamido, 2-(azidomethylthio)acetamido, 2-(ethylsul-
fonyl)acetamido, 2-(2,2,2-trifluoroethylthio)acetamido, ~
2-(methylsulfinyl)acetamido and 2-(cyanomethylsulfinyl)acet-
amido.
Illustrative of the group R3-C-NH- in the above
definition in which R3 is Y -CH2- are the following: 2-(2-
thienyl)acetamido, 2-(3-thienyl)acetamido, 2-(2-~uryl)acet-
amido, 2-(2-oxazolyl)acetamido, 2-(2-thiazolyl)acetamido,
2-(1-tetrazolyl)acetamido, 2-(1-benzotriazolyl)acetamido,
2-(2-oxazolylthio)acetamido, 2-(2-thiazolylthio)acetamido,
:
X-4902A -1~
'..''
,: ' '"
. - ~ .
.: :, : ' , . . '. '
. :,: ,,: .
3~4~
2-tl,2,3-triazol-5-ylthio)acetamido, 2-(1,3,~-triazol-2- ~ :
yl-thio)acetamido, (1,3,4-thiadiazol-2-ylthio)acetamido,
2-(~-(pr~tected amino)-ll3~4-thiadiazol-2-ylthio)acetamid
2-(5-methyl-1,3,4-thiadiazol-2-ylthio)acetamido, 2-(1,2,4-
thiadiazol-5-ylthio)acetamido, 2-(3-methyl-1,2,4-thiadiazol-
5-ylthio)acetamido), 2-(1,2,5-thiadiazol-3-ylthio)acetamido, ~.
2-(1,3,4-oxadiazol-2-ylthio)acetamido, 2-(5-methyl-1,3,4-
oxadiazol-2-ylthio)acetamido, 2-(1-methyl-5-tetrazolyl- ~
thio)acetamido, 2-(pyridylthio)acetamido, 2-(4-cyano-1,2,3- :~ -
triazol-l-yl)acetamido, 2-~3-cyano-1,2,4-triazol-1-yl)~
acetamido, and 2-(2-(protected amino)-4-thiazolyl)acet-
amido(tautomeric with 2-(2-imino-4-thiazolin-4-yl)acet-
amido).
Many of the cephalosporanic acids to be employed .
as starting material in the present process are known com-
pounds, and all are prepared by methods known to those .~ .. . .
skilled in the art. Attention is directed to "Cephalosporins
And Penicillins, Chemistry and Biology", edited by Edwin H.
Flynn (Academic Press, New York, 1972). See espe~ially
~0 Chapters 3, 4, and 15, and the references there cited. See
also U.S. 3,914,157 and South African Patent 71/3229.
The preferred sulfur nucleophiles o~ Formula III
are those wherein R is of the formula C r R6 .
Suitable fi~e-membered heteroaromatic rings of
said formula incluae the following:
pyrrole
oxazole
isoxazole . - .
thiazole .~. . .
isothiazole
pyrazole :: .
X-4902A -19-
j.''', '"."
.; ~_ , I .
. - ., . .. .. , . , . : . ,, , , .- .. . : ,
' " ~ " ','-''.''.'; ' ' ' ' ~" ~ ' '
~ 3~
imidazole
1,2,3-oxadiazole
1,2,4-oxadiazole
1,2,5-oxadiazole
1,3,4-oxadiazole
1,2,3-thiadiazole
1,2,4-thiadiazole
1,2,5-thiadiazole
1,3,4-thiadiazole
1,2,3-triazole
1,2,4-triazole
oxatriazole
tetrazole
Suitable six-membered rings include the following:
pyridine
pyridazine
pyrimidine
pyrazine
1,2,3-triazine
1,2,4-triazine
1,3,5-triazine
1,2,3,4-tetrazine
1,2,4,5-tetrazine
Certain of the heteroarylthiols, as well as
certain of the alkylthiols and phenylthiols of Formula III,
.
;~ actually exist as thiones or as tautomeric mixtures of thiol .
and~thione. As an example, the compound 2~methyl-1,3,4-
:: thiadiazole-5-thiol exists as a tautomer:
H
S 5=~ CH3
However, the present reaction proceeds with the designated
classes of sulfur nucleophiles regardless of whether the
: reactant is in the thiol or in the thione form. Accord- : :
~ ~ ,.... .
ingly, it is within the scope of the present invention to
conduct the reaction employing a thione or thiol-thione
.
~ : tautomer of a designated reactant.
~ ~ ' ,. .
:30 :
X 4902A ~ -20-
:
, . . . .
.
,. .. ~,
.. .. : . .,
' ' `., ': ' . , ' ' ' ' ` ,' , : ' ' ', ' . . ' ' ' ' ` ' , `
: , ' .
. .
The heteroarylthiol can be unsubstituted or can be
substituted by one or more substituents. In general, the
identity of substituents is not critical. For best results,
any primary or secondary amino group should be protected.
Where a heteroarylthiol contains more than one thiol group,
the one which is desired not to undergo reaction should be
protected. Methods for protection of thiols are well known;
see McOmie, supra. Assuming the protection of such groups,
the reaction goes forward regardless o~ the identlty of
substituent.
It is preferred to avoid excessively bulky sub-
stitution; generally, substitution not exceeding a molecular
weight of about 500 is preferred. Most common substituted
heteroaromatic rings contain substitution of a total molecular
weight less than about 250.
Suitable substituents include Cl-C4 alkyl, C2-C4
alkenyl, C2-C4 alkynyl, C5-C6 cycloalkyl, benzyl, phenethyl, -
aryl including phenyl and substituted phenyl as defined
hereinabove, halo -CF3, -O, -OH, protected amino or Cl-C4
alkylamino, -COOH, -COOA , -CONH2, protected aminomethyl or
Cl-C4 alkylaminomethyl, -CH2SO3-alkali metal, -CH2COO~
-CH2COOA r or -(CH2)2NtCH3)2-
Representative sulfur nucleophiles of Formula IIIinclude the ~ollowing:
Thioureas
allylthiourea
N,N'-di-n-butylthiourea
N,N'-di-tert-butylthiourea
N,N'-dicyclohexylthiourea
X-4902A -21-
. .
, , , . . :,
L4~ ~
N,N'-diisopropylthiourea :
N,N'-dimethylthiourea
N-methylthiourea
N,N,N',N'-tetraethylthiourea
N,N,N',N'-tetramethylthiourea
thiourea . -
N-phenylthiourea
N,N'-diphenylthiourea
N,N-dimethylthiourea
Thiobenzoic acid
Alkylthiols
methanethiol
ethanethiol
1- or 2-propanethiol
1- or 2-butanethiol
2-methyl-1-propanethiol
Benzenethiols :
benzenethiol ~.:.::
4-bromo-3-methylbenzenethiol . : .
p-bromobenzenethiol : ~ .
. p-chlorobenzenethiol
:~ 2,5-dichlorobenzenethiol :~
3,4-dichlorobenzenethiol ~
4-fluorobenzenethiol ~ :
m-methoxybenzenethiol :~:
p-methoxybenzenethiol ~ ~ :
p-nitrobenzenethiol ::
p-methylbenzenethiol
..
X-4902A -22-
3~41
Heteroar~lthiols
2-pyrrolethiol : -
3-pyrazolethiol
2-methyl-3-pyrazolethiol
2-methyl-4-imidazolethiol
2-imidazolethiol
4-oxazolethiol
3-isoxazolethiol
2-thiazolethiol
3-isothiazolethiol
l-methyl-1,2,3-triazole-5-thiol
l-benzyl-1,2,3-triazole-4-thiol
2-methyl-1,2,3-triazole-4-thiol
3,5-dimethyl-1,2,3-triazole-4-thiol
1,2j4-triazole-3-thiol ~ :
4-methyl-1,2,4-triazole-5-thiol
3-methyl-1,2,4-triazole-5-thiol :: .
,:,
3,4-dimethyl-1,2,4-triazole-5-thiol
2-methyl-1,2,4-triazole-5-thiol
2-benzyl--1,2,4-triazole-5-thiol ;
2,3-dimethyl-1,2,4-triazole-5-thiol .
4-methyl-3-(trifluoromethyl)-1,2,4-triazole-
5-thiol :.
3-carbamoyl-4-methyl-1,2,4-triazole-5-thiol
, . . . .
:~ 3-~carboxymethyl)-4-methyl-1,2,4-triazole-
5-thiol
3-(carboethoxymethyl)-4-methyl-1,2,4-tri-
azole-5-thiol
: protected 3-amino-4-methyl-1,2,4-triazole- ~.
5-thiol ::
3-hydroxy-4-methyl-1,2,4-triazole-5-thiol
X-4902A -23-
.
3~
5-methyl-1,2,3-oxadiazole-4-thiol
1,2,4-oxadiazole-3-thiol
3-methyl-1,2,4-oxadiazole-5-thiol
3-methyl-1,2,5-oxadiazole-4-thiol
2-methyl-1,3,4-oxadiazole-5-thiol
3-methyl-1,2,4-thiadiazole-5-thiol
1,3,4-thiadiazole-5-thiol
2-methyl-1,3,4-thiadiazole-5-thiol
2-(N-methylacetamido)-1,3,4-thiadiazole-5-
thiol
10
5-tetrazolethiol
l-methyltetrazole-5-thiol
l-benzyltetrazole-5-thiol ~:
l-(carboxymethyl)tetrazole-5-thiol ~ .
l-((sodiooxysulfonyl)methyl)tetrazole-5-
thiol
1-(2-dimethylaminoethyl)tetrazole-5-thiol :
1,2,3,4-oxatriazole-5-thiol
2-pyridinethiol
2-pyridinethiol, l-oxide
3-pyridazinethiol :
:. . .
~ : 2-pyrimidinethiol
.
2-pyrazinethiol -:
1,2,3-triazine-4-thiol
1,2,4-triazine-3-thiol
4,5-dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-
triazine-3-thiol ~ :
: 1,3,5-triazine-2-thiol .:
1,2,4,5-tetrazine-3-thiol .
2-benzimidazolethiol
2-benzothiazolethiol
X-4902A -24-
: : - .'. . ".' , ' ' . .; ~ .. :,
2-benzoxazolethiol
tetrazolo(l,5-b)pyridazine-6-thiol.
Most of the sulfur nucleophiles to be employed in
the present process are known compounds, and all can be
prepared in accordance with prior art procedures. In the
case of the heteroarylthiols, attention is directed to the
numerous volumes of "Heterocyclic Compounds", edited by
Robert C. Elderfield (John Wiley and Sons, Inc., N.Y.), as~-
well as the various volumes on the particular heterocyclic
systems in the series "The Chemistry of Heterocyclic Com-
pounds," edited by Weissberger et al. (John Wiley and Sons,
N.Y.).
It is critical to the present invention that the
reaction be carried out in an organic solvent. However, the
identity of the solvent is not critical, since a great
variety of organic solvents has proven satisfactory. In
general, members of the following classes of solvents have
been found satisfactory: hydrocarbons, both aliphatic and
aromatic, alcohols, amides, ethers, ketones, carboxylic
acids, carb~xylic acid esters, halogenated hydrocarbons,
nitro compounds, nitriles, and thioethers. Since certain of
the nucleophiles are liquids, an excess of such reactant can
also be employed as solvent.
The solvent should be inert, in the sense that it
does not undergo a competing reaction with either reactant.
Particular solvents which are suitable in carrying
out the present practice include pentane, cyclopentane,
hexane, heptane, octane, benzene, toluene, o-, m-, or p-
xylene, cumene, mesitylene, p-cymene, l-pentene, diethyl
X-4902A -25-
.
.- . ... . . : ,
ether, butyl ethyl ether, diamyl ether, benzyl ethyl ether,
acetone, methyl ethyl ketone, 2-butanone, 3-pentanone,
methyl isobutyl ketone, cyclohexanone, acetic acid, pro-
pionic acid, butyric acid, isobutyric acid, valeric acid,
methyl acetate, ethyl acetate, propyl acetate, isopropyl
acetate, ethyl propionate, butyl acetate, isobutyl acetate, ~ .
sec-butyl acetate, amyl acetate, isoamyl acetate, ethyl :~
isovalerate, methyl benzoate, benzyl acetate, methyl
butyrate, diethyl carbonate, dimethyl maleate, diethyl
oxalate, ethylene glycol diacetate, diethyl malonate,
fluorobenzene, chlorobenzene, bromobenzene, o-, m- or p-
fluorotoluene, o-, m-, or p-chlorotoluene, o-, m- r or p-
bromotoluene, 2-chloroethane, 1- or 2-chloropropane, 1- or
2-chlorobutane, 1-chloro-2-methylpropane, 1, 2, or 3-chloro- :
pentane, l-chloronaphthalene, methylene chloride, chloro- -
form, carbon tetrachloride, l,l-dichloroethane, 1,2- :
dichloroethane, l,l,l-trichloroethane, 1,1,2-trichloro-
ethane, 1,1,2,2-tetrachloroethane, o-, m-, or p-dichloro-
benzene, nitromethane, nitroethane, 1- or 2-nitropropane, ~ . -
nitrobenzene, acetonitrile, propionitrile, butyronitrile,
isobutyronitrile, valeronitrile, benzonitrile, phenyl-
acetonitrile, and thiophene.
Preferred solvents are: .
Hydrocarbons: ;: .
benzene
toluene
o-, m-, or p-xylene .
cumene
mesitylene --
X-4902A -26-
' ' :
. -: .- :, . . ., ~: . . . . .
: ~ . . : , :. .
' :., , ' ' . ., . i
33~
Halogenated Hydrocarbons~
chloroethane -. .
1- or 2-chloropropane
1- or 2-chlorobutane
isobutyl chloride
1,2, or 3-chloropentane
methylene chloride
chloroform
1,2-dichloroethane ~:~
carbon tetrachloride
l,l,l-trichloroethane ~ `
1,1,2-trichloroethane
1,1,2,2-tetrachloroethane
fluorobenzene
chlorobenzene
bromobenzene
o-, m-, or p-fluorotoluene :~
o-, m-, or p-chlorotoluene
o-, m-, or p-bromotoluene
. 20 o-, m-, or p-dichlorobenzene
; ~ Acids:
acetic acid
propionic acid
butyric acid
isobutyric acid
valeric acid
~. .
X-4902A -27-
,, , .. , ~ .
, ~ .,............ . ~ :, -:
': ,'' ' '.'' ' ' -, , ' , : . "'
3~
Esters
. _ :
methyl acetate
ethylene glycol diacetate :
ethyl acetate ~-
propyl acetate :
isopropyl acetate
butyl acetate
isobutyl acetate :-
sec-butyl acetate
pentyl acetate -: .
ethyl propionate
methyl butyrate
n-butyl formate
propylene carbonate ~-
ethylene carbonate
Nitro Com~ounds:
nitrobenzene
nitromethane
nitroethane
nitropropane :~
Nitriles:
;~ acetonitrile
: . propionitrile
butyronitrile
: isobutyronitrile :-
. ~ valeronitrile .:: -
..
benzonitrile : .
phenylacetonitrile
,
:~
X-4902A -28
,. ~ '
~' ~; ' '
. : : ~, ,,, . . .. ,. :: .. , ,, . . ~ .
~ 33~
Ketones: -
methyl isobutyl ketone
methyl ethyl ketone
Especially preferred solvents are acetonitrile,
1,2-dichloroethane, methylene chloride, propionitrile, -
nitromethane, nitroethane, acetic acid, isopropyl acetate,
butyl acetate, and methyl isobutyl ketone. ;
The 3-(acyloxymethyl)cephalosporin of Formula II
has been defined as the acid~ It is believed to be the acid
which undergoes reaction, since cephalosporin salts are
generally not soluble in organic solvents. The exception is
the solubility of cephalosporin salts in carboxylic acid
solvents, such as acetic acid. Therefore, the present
reaction can be accomplished by dissolving a 3-(acyloxy-
methyl)cephalosporin salt, such as a metal salt, in a
carboxylic acid solvent.
Regardless of the identity of the solvent, it is
necessary that the present process be conducted under
essentially anhydrous conditions. In general, the reaction
mixture should contain less than 5 percent of water, and
preferably less than 1 percent of water. It is even more
preferred that the amount of water be less than 0.5 percent.
Where commercial sources of reactants and solvents are not
dry enough, removal of water can be carried out in accor-
dance with known procedures, including azeotroping and the
use of drying agents such as alumina, silica gel, anhydrous
calcium sulfate, and the like.
::, ' '.
X-4902A -29-
-- .
~-~f33~
The present reaction goes forward under a wide
range of temperatures. In general, reaction temperatures of
50-140C. can be used; but better results are usually
achieved at temperatures of 70-120C. The reaction can be
conducted at elevated pressures, but no advantages have been
observed. Hence atmospheric pressure is generally preferred
because of its simplicity.
The amounts of the reactants are not critical. In
general, an excess of the sulfur nucleophile is preferred,
such as 1.0 to 5.0 molar equivalents of sulfur nucleophile
per molar equivalent of the 3-(acyloY,ymethyl)cephalosporin
reactant. :-
The present process is particularly useful for the
introduction into cephalosporins of the following 3-
heteroarylthio groups:
5-methyl-1,3,4-thiadiazol-2-ylthio-
l-methyl-lH-tetrazol-5-ylthio-
. .. ~ .
l-carboxymethyl-lH-tetrazol-5-ylthio-
4-methyl-6-hydroxy-5-oxo-1,2,4-triazin-
3-ylthio-
l-methyl-1,3,4-triazol-2-ylthio- ~ -
2-methyl-1,3,4-triazol-5-ylthio-
1,2-dimethyl-1,3,4-triazol-S-ylthio-
.
3-methyl-1,3,4-triazol-5-ylthio-
2,3-dimethyl-1,3,4-triazol-5-ylthio- ~
l-methyl-2-trifluoromethyl-1,3,4-triazol- ~ ~ `
5-ylthio-
l-methyl-2-carboxamido-1,3,~-triazol-
5-ylthio-
~-4902A ~ ~
." ',' .
, ,- .. . . . . . , , :
. - . . ~ ' '. ' -. ', ,, . ' ' ',. . . : . .
.. . . . .
3~4~L
l-methyl-2-carboxymethyl-1,3,4-triazol-5-ylthio-
l-methyl-2-carboalkoxy-1,3,4-triazol-5-ylthio- :~
. l-methyl-2-(protected aminomethyll-1,3,4-triazol-
5-ylthio-
l-methyl-2-(protected amino)-1,3,4-triazol-5-
ylthio-
l-methyl-2-hydroxy-1,3,4-triazol-5-ylthio-
lH-tetrazol-5-ylthio-
1-(2-dimethylaminoethyl)-lH-tetrazol-5-ylthio-
1-(sulfomethyl)-lH-tetrazol-5-ylthio-, alkali
metal salt
5-methyl-1,3,4-oxadiazol-2-ylthio-
1,3,4-thiadiazol-2-ylthio-
3-methyl-1,2,4-thiadiazol-5-ylthio- and
: 2-(protected aminomethyl)-1,3,4-thiadiazol
5-ylthio-.
Among the important known cephalosporin compounds
of Formula I whlch can be prepared by synthetic routes ;~
:~ incorporating the present process are the following:
l ~S ~:~
; ~ cefamandole~CH-C-NH-
H ~ CH2-S~
COOH ~
, -
` ~ .,:: :
, ,''',
~'; ;' '~'
X-4902A : -31-
., . .. .... . ~ .. . .
. . , , . , ~ .
L4~L
cefamandole O /S
~OH ~ /
cefazolin: ~ /S -
-CH -C-NH-~- t `~
o~ 2 ~ ~ CH3
OOH
ceftezole: ~ ~ /S~
l o ~ I_C~2_S_~
~OOH
O ',~' ~ ~' '' ', '
ce:Eazaflur: 11 /S\ - :~
~ CH -S~
- O S :~
SCE-963 F= - CH2-CNH~
S ~ / :
I OOH
NH2 CH -CH -N(CH ) . :
o
æNH 0~ t ~ CH -S- ~ N
O OCH
~ CS-1170: NC.H SCH CNH ~ N-----N
X-4902A -32-
~ 33~
SQ-67,590: NCCH SCH CNH-~
OOH CH
SKF-75073 ~ ~S\
~ /o-CH-CNH ~ t N-----N
OH O~ cH2-s-~
OOH ~H SO H
:,
O
BBS-226~ 11 / S \ N-- =N
-CH2-CNH-~ N--N /
CH2NH2 O~ ~ ~ CH2 S ~\ /3= = N , and
OOH
:: .
. : 20 ~ : ~ jS
SQ-14,359~ -CH-CNH-~ f ,N~
\ ~ \ N ~ .
C=O OOH CH3
: NH2
The products of the present inven~ion, their
deblocked derivatives, and the pharmaceutically acceptable
-:
salts of the same are useful in combating infections in
X-4902A -33-
.. :. - : - ~ ,. . ,
' ~ : . . ~ ; . , .:
: ' . : ,
~ 33~
warm-blooded mammals when administered parenterally in
non-toxic doses between about 10 and 500 mg./kg. of body
weight. The compounds are formulated in conventional
procedures.
The following examples illustrate the present
invention and will enable those skilled in the art to
practice the same.
EXAMPLE 1: PREPARATION OF 3-(((1-MET~YL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ACETONITRILE.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.2 grams; 5.5 mmole) and 1-methyl-lH-tetrazole-5-thiol
(1.0 gram; 8.6 mmole) were added to 25 ml. of acetonitrile
in a flask equipped with a condenser with a drying tube
containing anhydrous calcium sulfate sold under the trademark
Drierite. The reaction mixture was reflu~ed, with stirring, -
and the progress of the reaction was monitored by thin-
layer chromatography (TLC). After 90 minutes at reflux, TLC
showed about two-thirds cephalosporanic acid starting
material and one-third of the desired product. After 3
hours of reflux, TLC showed about one-third cephalosporanic
acid starting material and two-thirds product. After 6
hours, at which time TLC showed the reaction essentially
complete, the reaction mixture was cooled to room tempera-
ture and allowed to stand overnight. Solvent was removed on
a rotary evaporator, leaving as a residue a foam which was ;
dissolved in 10 ml. of ethanol. Dropwise addition of 1 ml.
of dicyclohexylamine in 10 ml. of ethanol resulted in the
precipitation o~ product as the dicyclohexylamine salt
X-4902A -34-
.. :, - . ., , .: . ........................ . . ..
" '. ' ''' , .' , ' ' ,. ' . ' , ' , ," ' ' '' :' . ': "
3~
which, after 15 minutes of stirring, was separated by
filtration. The isolated product was washed with 15 ml. of
ethanol, and the product was vacuum dried at 40C. for 2
hours, 2.50 grams (76.1 percent yield), m.p., 183-4C.
(dec). The product was subjected to NMR, IR, and TLC, and
all of these analyses were identical with analyses of an
authentic sample of the product prepared by the prior art
aqueous displacement. NMR(DMSO-d6) ~ 3.52 (m, 2, 2-CH2),
3.76 (s, 2, -CH2CONH-), 3.92 (s, 3, -CH3 of tetrazole), 4.35
(s, 2, 3-CH2S), 5.00 (d, 1, C6-H, J=5 Hz), 5.55 (q, 1,
C7-H, J=5 Hz, J=9 Hz), 6.95 (d, 2, thiophene 3 and 4-H, J=3
Hz), 7.35 (t, 1, thiophene 5-H, J=3 Hz), and 8.75 (d,
1, -CH2CONH-, J=9 Hz).
EXAMPLE 2: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL~THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ACETONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(4.0 grams; 10 mmole) and 1-methyl-lH-tetrazole-5-thiol (5.8
grams; 50 mmole) were refluxed in 50 ml. of dry acetonitrile
(treated over a anhydrous sulfonic acid resin sold under the
trademark Amberlite 15), under a dry atmosphere, for 8 3/4
hours. The reaction was followed by 'rLC and was nearly
complete at the end of 8 3/4 hours.
Solvent was removed by evaporation and the residue
added to 125 ml. of ethanol. Dicyclohexylamine (6 ml.) in
75 ml. of ethanol was added and the product precipitated as
the dicyclohexylamine salt. It was separated by filtration,
washed, and dried! 4.50 grams (71.0 percent yield). IR,
NMR, and TLC were identical to an authentic sample prepared
X-4902A -35-
. , -,' ~, ,'',., .. , , ' .~ , .' ' . " . :
.. . . . . . . . . . . . . . .
- .. : , . - . ,; :
.. , ,.......... : , . .
~;)8319~
by aqueous displacement. The NMR was also identical wlth
the NMR on the product of Example l. ~; -
EXAMPLE 3: PREPARATION OF 3-(((l-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (1.2
grams; 10 mmole) in 25 ml. of 1,2-dichloroethane were
refluxed under a dry atmosphere for a ~otal of 5.5 hours.
Solvent was then removed on a rotary evaporator, and 25 ml.
of ethanol was added to the filtrate followed by dropwise ~-
addition of a solution of 2 ml. of dicyclohexylamine in ~ ;~
25 ml. of ethanol. The product crystallized as the dicyclo- ~
hexylamine salt and was stirred ~or 20 minutes at room -
temperature, then filtered, washed with 25 ml. of ethanol, -
and dried at 40C. under vacuum, 2.34 grams of off-white
solid (73.8 percent yield). IR and NMR were identical with
an authentic sample prepared by aqueous displacement. The
NMR was also identical with the NMR on the product of
Example 1.
EXAMPLE 4: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-AMINO-3-CEPHEM-4-CARBOXYLIC ACID IN 1,2-
DICHLOROETHANE
7-Formamidocephalosporanic acid (3.0 grams; 10
mmole) and l-methyl-lH-tetrazole-5-thiol (2.4 grams; 20
mmole) in 50 ml. of 1,2 dichloroethane were stirred and
refluxed for 7 hours. The reaction mixture was then cooled
to room temperature and allowed to stand overnight at room ~-
temperature. A red gummy solid precipitated. The product
was identified by removing the 7-formyl group as follows.
X-4902A -36-
.
:, ' ' ' "
, .
',. '' '.~' ', '' ' " '''''.', '", '' ''" '","''' ' ., "' ' ~ "
.. ......
: . - , .
. . . . . . . . . .
~Lafl3~
Solvent was removed on a rotary evaporator and the
residue was dissolved in 25 ml. of methanol and 2.8 ml. of
concentrated HCl and permitted to stand overnight at room
temperature. The solution was then diluted to 50 ml. with
water, and the pH, initially 0.9, was raised to 3.6 by
dropwise addition of triethylamine. Light brown crystals
~ere filtered, washed with water, and dried, 2.10 grams (64
percent yield). The NMR was identical with the same product
made by aqueous displacement. NMR(D2O, NaHCO3) ~ 3.65 (q,
2, 2-CH2, JAB=16.5 Hz), 4.08 (s, 3, -CH3 on tetrazole), 4.16
(q, 2, 3-C_2S-, JAB=12.5 Hz), 5.05 (d, 1, C6-H, J=5 Hz), and ~-
5.45 (d, 1, C7-H, J=5 Hz).
EXAMPLE 5: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
~L)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ISOPROPYL ACETATE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (1.2
grams: 10 mmole) in 25 ml. of isopropyl acetate were refluxed
(90C.) with stirring for 23 hours.
The reaction mixture was then cooled to room
temperature. TLC showed some of the cephalosporanic acid
starting material remaining. The light cream colored solid
was separated by filtration, washed with isopropyl acetate
and dried, 1.60 grams (70.8 percent yield). NMR confirmed
the identity of the product and showed less than 1 percent
of the cephalosporanic acid starting material present. NMR
(DMSO-d6) ~ 3.72 (s, 2, 2-CH2), 3.80 (s, 2, -CH2CONH-),
3.95 (s, 3, -CH3 of tetrazole), A.30 (s, 2, 3-CH2S-), 5.10
(d, 1, C -H, J=5 Hz), 5.70 (q, 1, C -ll, J=5 llz, J=8 Hz),
6 - - 7 - - -
X-4902A -37-
:', .'', . ' ~, ': , "
~38314~
6.92 td, 2, thiophene 3- and 4-H, J=3 Hz), 7.35 (t, 1, i
thiophene 5-H, J=3 Hz), and 8.78 (d, 1, -CH2CONH-, J=8 llz).
EXAMPLR 6: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CA~BOXYLIC ACID IN PROPIONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (1.2
grams; 10 mmole) were refluxed in 25 ml. of dry propioni-
trile (97C.) until TLC showed the complete disappearance of
cephalosporanic acid starting material (4.5 hours). The
reaction mixture was then cooled to room temperature, and
solvent was removed on a rotary evaporator, during which
process some of the solution was lost due to bumping. The
residue was dissolved in 35 ml. of warm ethanol and a
solution of 2 ml. of dicyclohexylamine in 10 ml. of ethanol
was added. The product precipitated as the dicyclohexyl-
amine salt and was stirred 10 minutes at room temperature,
then filtered, washed with ethanol and dried, 1.24 grams
(39.0 percent yield, not including product lost on the
rotary evaporator). The NMR of the product was identical
with the NMR on the Example 1 product.
EXAMPLE 7: PREPARATION OF 3-(((l-METH~L-lH TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ACETONITRILE, 3,5-DXCHLOROPHEN~L DIHYDROGEN
PHOSPHATE ADDED
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole), 1-methyl-lH-tetrazole-5-thiol (0.87
gram; 7.5 mmole), and 3,5-dichlorophen~l dihydrogen phosphate
(0.122 grams; 0.5 mmole) were heated overnight in 25 ml. of
X-4902A 38-
.
dry acetonitrile at 70C. Solvent was then removed on arotary evaporator to 8-10 ml. The product began to crys-
tallize. After stirring for 1/2 hour, isopropyl acetate (25
ml.) was added dropwise during 1/2 hour to effect further ~-
crystallization. After an additional 1/2 hour of stirring,
the product was separated by filtration, washed with
isopropyl acetate and dried, 0.98 gram (43.4 percent yield).
The NMR of the product was identical with the NMR on the
Example 5 product.
EXAMPI.E 8: PREPARATION OF 3-(((lH-TETRAZOL-5-YL)THIO)-
METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID IN ACETIC ACID
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.87
grams; 7~5 mmole) were added to glacial acetic acid (25
ml.). The reaction mixture was heated to 60~C. and held at
this temperature for an hour. Only a trace of product was
evident on TLC. The reaction mixture was heated to 80C.
and held at this temperature for 5 hours. The reaction
mixture was then allowed to stand overnight, and the product
was separated by filtration, washed with acetic acid, and
dried, 0.71 gram (31 percent yield). The NMR of the product
was identical with the NMR on the Example 5 product.
EXAMPLE 9: PREPARATION OF 3 (((4,5-DIHYDRO-6-HYDROXY-4-
METHYL-5-OXO-1,2,4-TRIAZIN-3-YL)THIO)METHYL)-7 (2-(2-
THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN ACETONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 4,5-dihydro-6-hydroxy-4-methyl-
5-oxo-1,2,4-triazine-3-thiol (1.2 grams; 7.5 mmole) were
X-4902A -39-
~8319L~L
added to 25 ml. of dry acetonitrile in a reaction flask
immersed in an oil bath at 84-85C. The flask was equipped
with a condenser with a drying tube containing anhydrous ~ -
calcium sulfate sold under the trademark Drierite. The
reaction mixture was maintained for 16 hours in this con- -
dition, with slow magnetic agitation.
The product crystallized from the hot solution.
The reaction mixture was cooled to room temperature, filtered,
washed with acetonitrile, washed with acetone, and vacuum
10 dried, 1.81 grams (72.6 percent yield). The product was -
off-white crystals, m.p. 161C. (dec).
On standing, the filtrate deposited a second crop
of crystals. This crop was separated by filtration, washed
with acetonitrile, and dried, 0.26 gram (10.5 percent
yield). This second crop also melted at 161C. (dec).
Total yield was therefore 83.1 percent.
EXAMPLE 10: PREPARATION OF 3-(((5-METHYL-1,3,4-OXADIAZOL-
2-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
~2.0 grams; 5 mmole) and 5-methyl-1/3,4-oxadizaole-2-thiol
(0.87 gram; 7.5 mmole) in 25 ml. of 1,2-dichloroethane were
placed in a flask equipped with magnetic stirring bar and
condenser with a drying tube containing anhydrous calcium
sulfate sold under the trademark Drierite. The flask was
immersed in an oil bath at 84-85C. and held for B hours,
then refrigerated for two days.
A portion of product had crystalli~ed; it was
separated by filtration, washed with 1,2-dichloroethane, and
30 dried, 1.10 grams (48.7 percent yield). It was dissolved in
X-4902A -40-
~:- .. , ~ ~ .
3~4'1
15 ml. of acetone and filtered to remove an insoluble
product. Then 75 ml. of deionized water was added dropwise
and the product was separated by filtration and dried, 0.61
gram, m.p. >114C. (dec.).
Evaporation of solvPnt from the filtrate yielded
0.23 gram of a mixture of product and both starting materials.
EXAMPLE 11: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-t2-t2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN NITR~METHANE
7-t2-t2-Thienyl)acetamido)cephalosporanic acid
t2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.87
gram; 7.5 mmole) were added to 25 ml. of dry nitromethane in
a flask; the flask was immersed in an oil bath at 100-101C. --
for 4.5 hours, then cooled to room temperature. TLC showed
that the reaction was complete.
Solvent was removed on a rotary evaporator, and
the residue was dissolved in 75 ml. of warm ethyl acetate.
The solution was filtered to remove a small amount of -
insolubles, then extracted with two 25-ml. portions of 5
percent aqueous sodium bicarbonate. The combined extracts
were layered with 50 ml. of ethyl acetate and acidified to
pH about 1.0 with 70 percent a~ueous methanesulfonic acid~
The ethyl acetate layer was separated and the water layer
was extracted with 25 ml. of ethyl acetate. The ethyl
; acetate layers were combined, dried over anhydrous sodium
sulfate, and concentrated to 25 ml. on a rotary evaporator.
Dropwise addition of 50 mlO of diethyl ether resulted in
crystallization of the product. It was separated by fil- -
X-4902A -41-
. .
-, ' ' : .: ' '
~L~)fl31~
tration, washed with diethyl ether, and dried, 1.27 gram
(56.2 percent yield), off-white crystals, m.p. 156-159C.
(dec.). The identity of the product was confirmed by NMR.
EXAMPLE 12: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-TEIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN METHYLENE CHLORIDE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(11.9 grams; 30 mmole) and 1-methyl-lH-tetrazole~5-thiol
(7.0 grams; 60 mmole) in 300 ml. of methylene chloride
(cyclohexane stabilized) were placed in a one-liter stainless
steel autoclave equipped with heater. The reaction mixture
was stirred and heated to 83-86C., developing a pressure of
42 p.s.i., for 16 hours. The reaction mixture was then
cooled to room temperatureO TLC showed conversion to
product, with only a trace of the starting cephalosporanic
acid remaining. The reaction mixture was permitted to stand
at room temperature, which resulted in crystallization of
the product, and then concentrated to 200 ml. The crystals
~were filtered off and washed with methylene chloride, 7.37
grams (54.3 percent yield), white crystals, m.p. 163.5-164C.
(dec.).
A second crop of light tan crystals was obtained
by diluting the filtrate with 100 ml. o~ diethyl ether,
filtering, washing with diethyl ether, and drying, 1.40
grams (10.3 percent yield).
;~ A third crop was obtained by diluting the filtrate
with isopropyl acetate, 1.18 grams (8.7 percent yield).
Total yield was therefore 73.3 percent.
X-4902A -42-
... . . . . .
,
-
EXAMPLE 13: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN FLUOROBENzENE
7-(2-(2-Thienyl)ace~amido)cephaIosporanic acid
(2.0 grams; 5.04 mmole) and 1-methyl-lH-tetrazole-5-thiol
(1.2 grams; 10 mmole) in 75 ml. of fluorobenzene (b.p.
85.1C.) were mixed in a flask equipped with a condenser
with a drying tube containing anhydrous calcium sulfate sold
under the trademark Drierite. The mixture was heated to
reflux and progress of the reaction was followed by TLC in
7/1 ethyl acetate/acetic acid. The reaction, which remained
heterogeneous throughout, was essentially complete in 72
hours.
The reaction mixture was then cooled to room
temperature and filtered to separate the precipitated
product. Th~ product was washed with fluorobenzene and
vacuum dried at 40C. for five hours, 2.13 grams (93.4
percent yield), off-white crystals, 161-162C. (dec).
Identity of the product was confirmed by NMR.
EXAMPLE 14: PREPARATION OF 3-(((1-METHYL-lH-TETRA~OL-5-
YL?THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN THIOPHENE
- 7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.87
gram: 7.5 mmole) in 25 ml. of ~hiophene were refluxed for 7
hours, during which time the product crystallized. The
reaction mixture was cooled to room temperature, stirred for
30 minutes to complete crystallization, and filtered. The
separated product was washed with 0~5 ml. of thiophene,
X-4902A -43-
." -
.. . . . .. ... . . ..
, - ., . . :
.
.
3~1
then vacuum dried for 2 hours, 1.82 grams (80.2 percent
yield), white crystals, m.p. 162-163C. (dec). TLC showed
that the filtrate contained additional product.
EXAMPLE 15: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-l~YL)ACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN ACETONITRILE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (O.76 gram; 2 mmole) and 5-methyl-1,3,4-thiadiazole-
2-thiol (O.33 gram; 2.5 mmole) in 10 ml. of reagent grade
acetonitrile were refluxed for two hours and forty minutes.
The product crystallized. The reaction mixture was cooled
in an ice bath and filtered to separate the product, which
was washed with three ml. of acetonitrile and dried at 40
under vacuum, 0.61 gram (67 percent yield). The identity of
the product was confirmed by NMR, ~ 2.68 (s, 3, -CH3 of
tetrazole), 3.72 (s, 2, 2-CH2), 4.40 (q, 2, 3-C_2S-, JAB=13
Hz), 5.12 (d, 1, C6-H, J=5 Hz), 5.38 (s, 2, -CH2CONH-),
5.72 (q, 1, C7-_, J=5 Hz, J=9 Hz), 9.00 (s, 1, tetrazole
5-H), and 9.17 (d, 1, -CH2CONH-). -
20 EXAMPLE 16: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-FORMYLOXY-2-PHENYLACETAMIDO)-3-CEPHEM-
4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2 Formyloxy-2-phenylacetamido)cephalosporanic
acid (2.17 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-
thiol (0.87 grams; 7.5 mmole) were added to 25 ml. of 1,2-
dichloroethane, and the resulting reaction mixture refluxed
for six hours. Samples were taken for TLC at 3 and 6 hours.
The reaction mixture was heated for an additional hour, then
allowed to cool overnight.
X-4902A -44-
'
8;~14~
Solvent was removed by evaporation. Diethyl
ether was added to the residue, which turned gummy and then
solid. The product was separated by filtration, washed with
diethyl ether, and dried, 1.90 grams (77.0 percent yield).
NMR(DMSO-d6) ~ 3.52 (s, 2, 2-CH2), 3.88 (s, 3, -CH3 of
tetrazole), 4.10 (s, 2, 3-CH2S-), 4.92 (d, 1, C6-H), 5.62
(q, 1, C7-H, J=5 Hz, J=9 Ha), 6.06 (s, 1, -CHCONH-), 7.26
(s, 5, phenyl-H), and 8.28 (s, 1, -OCH).
EXAMPLE 17: P~EPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
' '
YL)THIO)METHYL)-7-(5-CARBOXY-5-(2,4-DICHLOROBENZAMIDO)-
VALERAMIDO) 3-CEPHEM-4-CARBOXYLIC ACID IN (A) ACETONITRILE
AND (B) 1,2-DICHLOROETHANE
(A) 7-(5-Carboxy-5-(2,4-dichlorobenzamido)valer-
amido)cephalosporanic acid (2.9 grams; 5 mmole), l-methyl-
lH-tetrazole-5-thiol tl.2 grams; 10 mmole), and 50 ml. of
acetonitrile were refluxed overnight (18 hours). TLC `
showed reaction had taken place, estimated to be about 90
percent complete. The reaction mixture was evaporated and
the residue slurried in ethyl acetate and filteretl, 1.51
grams (48.3 percent yield).
(B) 7-(5-Carboxy-5-(2,4-dichlorobenzamido)valer-
amido)cephalosporanic acid (2.9 grams; 5 mmole), 1-methyl-
lH-tetrazole-5-thiol (1.2 grams; 10 mmole) and 50 ml. of
1,2-dichloroethane were refluxed for five and one-half
hours. TLC showed that reaction was 90 percent complete.
Solvent was decanted from the reaction mixture,
leaving a plastic-like solid which was triturated under
refluxing ether. The product was separated by filtration,
X-4902A -45
, ... .
- - .. . . . . . . .
- ~ . ~ . ' ,,
. --., '. . ' `, ' , , i :
'. ,,
2.83 grams (89.3 percent yield), tan crystals. NMR corrobo-
rated the identity of the product but showed ether. NMR
(DMSO-d6) ~ 1.78 and 2.26 (each m, adipoyl side chain), 3.66
(m, 2, 2-CH2), 3.95 (s, 3, -CH3 of tetrazole), 4.30 (m, 2,
3-CH2S-), 5.08 (d, 1, C6-H, J=5 Hz), 5.68 (q, 1, C7-H, J=5
Hz), 7.50 and 7.62 (each s, 2,4-dichlorophenyl), and 9.00
(m/ 2, two -CONH).
EXAMPLE 18: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(0.99 gram; 2.5 mmole) and 1-methyl-lH-tetrazole-5-thiol
(0.58 gram; 5 mmole) were added to 12.5 ml. of 1,2-dichloro-
ethane and the reaction mixture was refluxed. The reflux
was conducted to permit removal of the acetic acid by-product;
this was accomplished by returning the 1,2-dichloroethane
reflux through calcium oxide.
After 5 3/4 hours of reflux, the reaction mixture
was cooled to room temperature and filtered. The fluffy
needles were washed in 10 ml. of 1,2-dichloroethane and
dried, 0.77 grams (68.1 percent yield).
Evaporation of the filtrate gave another 0.04 gram
of product (3.5 percent yield).
The identity of the product was confirmed by TLC
and, in the case of the main product, by NMR. The NMR was
identical with the NMR on the Example 5 product.
X-4902A -46-
': - ' ' ' ' ' " ', ' '. ' ' , '
EXAMPL~ 19: PREPA~ATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIC)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ACETONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 5-methyl-1,3,4-thiadiazole-2-
thiol (2.64 grams; 20 mmole) in 25 ml. of acetonitrile were
refluxed overnight at 79C. TLC showed only a trace of
cephalosporanic acid starting material. The reaction mixture
was filtered, to remove an oily residue, and solvent was
removed on a rotary evaporator. The residue was crystallized
from 1:1 acetonitrile:isopropyl acetate, separated by
filtration, washed and vacuum dried, 1.59 gram (33.9 percent
yield) m.p. 166C. The identity of the product was confirmed
by IR, UV, NMR, MS, and elemental analyses. NMR (DMSO-d6)
2.68 (s, 3, -C_3 of tetrazole), 3.68 (s, 2, 2-CH2), 3.76
(s, 2, -C_2CONH-), 4.38 (q, 2, 3-CH2S-, J-13 Hz), 5.10 (d,
lj C6-H, J=5 Hz), 5.70 (q, 1, C7-H, J=5 Hz, J=9 Hz), 6.92
(d, 2, thiophene 3- and 4-H), 7.37 (t, 1, thiophene 5-H),
and 9.10 (d, 1, -CH2CONH, J=9 Hz).
20 EXAMPLE 20: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)~CETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,1,2-TRICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (1.2
grams; 10 mmole) were added to 25 ml. of 1,1,2-trichloro-
ethane and heated to 100-1C. The reaction was followed by ~ ;
TLC; after 4 hours at 100-1C., with stirring, TLC showed no
cephalosporanic acid starting material remaining. The
reaction mixture was cooled to room temperature, with stirring.
X-4902A -47-
.
.. .. , ~ ,
. : . .. , ,. ; , . . ... . . .
, , , , , , ' ' ~
A seed was introduced and the reaction mixture was allowed
to stir overnight. The product crystallized. Solvent was
removed by evaporation and 25 ml. of 1,2-dichloroethane
added. Solvent was again removed by filtration, and the
product was washed and vacuum dried.
There was obtained 0.98 gram of 3-(((l-methyl-
lH-tetrazol-5-yl)thio)methyl)-7-(2-(2-thienyl)acetamido)-
3-cephem-4-carboxylic acid, m.p. 158C (dec) (43.4 percent -~
yield).
EXAMPLE 21: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACBTAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN (A) METHYL ETHYL KETONE AND (B) 1,1,2-
TRICHLOROETHANE
(A~ 7-(2-(2-Thienyl)acetamido)cephalosporanic
acid (2.0 grams; 5 mmole) and l-methyl-lH-tetrazole-5-
thiol (1.2 grams; 10 mmole) were added to 25 ml. of methyl
ethyl ketone and refluxed 48 hours. TLC showed that only a
trace of cephalosporanic acid starting material remained.
The reaction mixture was washed with a mixture of 2.5 grams
of sodium bicarbonate in 50 ml. of water. The water layer
was treated with l gram of carbon and 50 ml. of ethyl
acetate was added. The pH was then lowered to pH 1.6 with
4 ml. of methanesulfonic acid in 30 ml. of water. The ethyl
acetate layer was dried over sodium sulfate and evaporated
to an oil on a rotary evaporator.
The product was crystallized by dropwise addition
of 50 ml. of ether. The product was separated by filtration,
washed, and vacuum dried, 0.94 gram (41.6 percent yield).
The NMR was identical with the NMR of the product of Example ~;~
5.
X-4902A -48-
, ' .;
~ 533~
(B) The reaction was repeated with the samestarting materials and amounts as in paxt (A), except that
50 ml. of 1,1,2-trichloroethylene was used as solvent. The
reaction mixture was refluxed for 16 hours, at which time
TLC showed that no cephalosporanic acid starting material
was present. The product was separated as described in part
(A), 0.47 gram (20.8 percent yield). The NMR was identical
with the NMR of the product of Example 5.
EXAMPLE 22: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-PHENYLACETAMIDO)-7-~THOXY-3-CEPHEM-
4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-Phenylacetamido)-7-methoxycephalosporanic
acid (210 mg.; 0.5 mmole), 1-methyl-lH-tetrazole-5-thiol (87
mg.; 0.75 mmole) and 15 ml. of 1,2-dichloroethane were mixed
and refluxed for 6 hours under nitrogen, at which time TLC
showed only a trace of cephalosporanic acid starting material.
Additional l-methyl-lH-tetrazole-5-thiol (29 mg.; 0.25
mmole) was added and the reaction mixture refluxed for an ;~ ;
additional three hours. TLC showed no appreciable change.
The reaction mixture was washed four times with
saturated sodium bicarbonate, and the bicarbonate layer was
washed three times with ethyl acetate, an amount of ~resh
ethyI acetate added, cooled to 0C. and adjusted to pH 2.2
with 20 percent HCl. The layers were separated, and the ~;
aqueous layer washed with ethyl acetate. The acetate layers
were combined, washed with saturated NaCl, dried over mag-
nesium sulfate, filtered, and evaporated, leaving a pale
green foam as a residue, 199 mg. (83.6 percent yield). The
identity of the product was confirmed by TLC and NMR.
X-4902A -~9- ~
'~.
.:. - ~,. . . . .. ,,:
. . . .
.. . . . ..
.
NMR(CDC13 + ld acetone-d6) ~ 3.45 (s, 3, -OCH3), 3.55 (s, 2,
2-CH2), 3-75 (s, 2, 0CH CO-), 3.9 (s, 3, CH3 on tetrazole),
4.4 (s, 2, 3-CH2S-), 5.15 (s, 1, C6-H), 7.35 (s, 5, 0), 8.0
(s, 1, -CH2CONH-), and ~ 11.2 (s, 1, J=o Hz, -COOH).
EXAMPLE 23: PREPARATION OF 3-((~2-BENZOTHIAZOLYL)THIO)-
METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(1.0 gram; 2.5 mmole) and 2-mercaptobenzothiazole (0.625
gram; 3.75 mmole) were placed in a flask, the air displaced
with a head of nitrogen, 25 ml. of 1,2-dichloroethane added,
and the reaction mixture heated to reflux and refluxed for
about 24 hours, with stirring. The reac~ion mixture was
then cooled and filtered, yielding the product as an off-
white material, 1.1 grams (88 percent yield), m.p. 190.5-191C.
(dec). It was dried overnight at 40C., under vacuum, and
submitted for analyses. The identity of the product was
confirmed by NMR, UV, mass spectroscopy, IR, and elemental
analysis. NMR(DMSO-d6) ~ 3.74 (s, 2, 2-CH2), 3.80 (s, 2,
20 -CH2CONH-), 4.58 (q, 2, 3-CH2S-, J=13 Hz), 5.14 (d, 1, ;
C6-H, J=5 Hz), 5.73 (q, 1, C7-H, J=5 Hz, J=8 Hz), 6-96
7.43, and 7.96 (each m, phenyl and thienyl rings), 9.10 (d,
1, -CH2CONH-, J=8 Hz)~
EXAMPLE 24: PREPARATION OF 3-(((5-(N^METHYLACETAMIDO)
: . , ,
1,3,4-THIADIAZOL-2-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACET-
AMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienylacetamido)cephalosporanlc acid (2.0
grams; 5 mmole) and 5-(N-methylacetamido)-1,3,4-thiadiazole-
2-thiol (1.42 grams; 7.5 mmole) were added to 50 ml. of 1,2-
X-4902A -50-
~ ~ ~ ' ' ' ' ' :, . '
3~
dichloroethane and the reaction mixture refluxed for 10
hours. The reaction mixture was cooled, filtered, washed
with 1,2-dichloroethane, and vacuum dried, 2.06 grams (78.3
percent), m.p. 178-9C. The identity of the product was
confirmed by IR, W, NMR, mass spectroscopy, and elemental
analysis. NMR(DMSO-d6) ~ 2.42 (s, 3, -COC_3), 3.74 (m, 7,
2-CH2, -CH3 of tetrazole, and CH2CONH ), 4.35 (q, 2, 3-C_2,
J=13 Hz), 5.10 (d, 1, C6-H, J=5 Hz), 5.70 (q, 1, C7-H~ -
J=5 Hz, J=9 Hz), 6.96 and 7.36 (each m, 3, thiophene H), and
9.10 (d, 1, -CH2CONH-, J=9 Hz).
EXAMPLE 25: PREPARATION OF 3-(~ METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(N-tert BUTOXYCARBONYL)-2-PHENYLGLYCYL-
AMIDO)-3-CEPHEM-4 CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
1,2-Dichloroethane (45 ml.) was refluxed to remove ;~
water-solvent azeotropes, then distilled down to 30 ml.,
which was permitted to cool. 7-(N-tert-Butoxycarbonyl-
2-phenylglycylamido)cephalosporanic acid (253 mg.; 0.5
mmole) was added and the solution distilled further, down to
15 ml. and again cooled. 1-Methyl-lH-tetrazole-5-thiol (87
mg.; 0.75 mmole) was added and the reaction mixture refluxed
under nitrogen and monitored by TLC. At 16 hours, TLC
showed only a trace of starting material.
The reaction mixture was washed four times with
saturated sodium bicarbonate and the bicarbonate layers ~
combined and washed twice with ethyl acetate. To the washed -
bicarbonate layers, fresh ethyl acetate was added, and the
solution was cooled to 0C. and adjusted to pH 2.4 with 20
percent HCl. The layers were separated. The aqueous layer
was washed with ethyl acetate.
X-4902A -51-
.~. ".. ...
~-
' ~ ' : , ' ,
. , . ' ~ . .. ' ,,
3~4: L
The ethyl acetate layers were combined, washedwith saturated NaCl, dried over magnesium sulfate, filtered
and evaporated, yielding a white foam, 224 mg. (80 percent
yield). The identity of the product was confirmed by TLC
and NMR. NMR showed a small amount (<.10 percent) of the
cephalosporanic acid starting material.
The product was suspended in 10 ml. of diethyl
ether and triturated for 1 hour. The ether was decanted,
and 10 ml. of fresh diethyl ether added and triturated for 1
hour. The ether was decanted and evaporated to dryness,
yielding the product as a white powder. The identity of the
product was confirmed by TLC and NMR. NMR (CDC13) ~ 1.45
(s, 9, -COO tert C4Hg), 3.6 (s, 2, 2-CH2), 3.95 (s, 3, CH3
on tetrazole), 4.3 (s, 2, 3-CH2S-), 4~9 (d, 1, J=6 Hz,
C6-H), 5.4 (d, 1, J=8 Hz, 0C_-), 5.75 (q, 1, J=4 Hz, C7-H), -
6.2 (d, 1, J=8 Hz, 0CH-), 7.4 (s, 5, 0), 7.65 (d, 1, J-8
NH
Hz, -CON_), and ~ 9.3 (s, 1, COOH).
EXAMPLE 26: PREPARATION OF 3~ 1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(N-(tert-BUTOXYCARBONYL)-2-(p-HYDROXY-
PHENYL)GLYCYLAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN 1,2-
DICHLOROETHANE
1,2-Dichloroethane (45 ml.) was refluxed with a
trap and 15 ml. solvent removed. The remaining 30 ml. was
cooled. 7-(N-(tert-Butoxycarbonyl)-2-(p-hydroxyphenyl)-
glycylamido)cephalosporanic acid (260.5 mg.; 0.5 mmole) was
added and the solution distilled further, down to 15 ml. and
cooled again. l-Methyl-lH-tetrazole-5-thiol (87 mg.; 0.75
mmole) was added and the reaction mixture heated to 65-70C.
X-4902A -52-
. . .
.
3~
and followed by TLC. TLC showed almost no starting material
after 3 hours, and the reaction mix~ure was worked up at the
end of 4 hours.
The workup and purification was essentially the
same as described in the preceding example. The yield was
189 mg. of crude product (65.5 percent) and 46 mg. of
purified product (about 16 percent). NMR(CDC13 + 2d acetone
d6) ~ 1.45 (s, 9, -COOtertC4Hg), 3.35 (s, 2, 2-CH2), 3.85
(s, 3, CH3 on tetrazole), 4.3 (s, 2, 3-CH2S-), 4Og (d, 1,
J=6 Hz, C6-H), 5.3 (q, 1, J=3 Hz, C7-H), 5.4 (s, 1, HO-0CH ),
5.75 (d, 1, J=6 Hz, HO0CH-), 6.4 (s, 1, COOH), 6.8 (d, 2, ~-~
N_
J=8 Hz, HO-~ H-), and 7.9 (d, 1, -CON~
~ . ;.: . .
EXAMPLE 27: PREPAR~TION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL~7-(2-PHENOXYACETAMIDO)-3 CEPHBM-4-CARBOXYLIC
ACID IN 1,2-DICHLOROETHANE
1,2-Dichloroethane (60 ml.) was refluxed with a `
trap and 15 ml. of solvent removed, leaving 45 ml., which
was cooled. 7-(2-Phenoxyacetamido)cephalosporanic acid (406
'~
mg.; 1 mmole) was added and the solution distilled further,
down to 30 ml. and cooled again. 1-Methyl-lH-tetrazole-5-
thiol (174 mg.; l.5 mmole) was added and the reaction
mixture was refluxed under nitrogen fox 12 hours, at which
time TLC in 10:3 diethyl ether/(3:1 acetic acid/water)
showed product, excess thiol reactant, a little cephalosporanic
acid reactant, and an unknown. ~
The reaction mixture was then worked up in essentially -
the same procedures reported in the preceding two examples.
X-4902A -53-
, ,' '' ' ' ' :, . , , , ,. ,. ' : ,'
.: . , ,, : - . , ., . , . ,, :
. .
3~
TLC of the final product showed it to be identical with an
authentic sample of product prepared by prior art methods.
NMR (CDC13) ~ 3.65 (s, 2, 2-CH2), 3.9 (s, 3, CH3 of tetrazole),
4-35 (s, 2, 3-CH2S-), 4.65 (s, 2, 0OCH2-), 5.1 (d, 1, J=4
Hz, C6-H), 5.9 (q, 1, J=4 Hz, C7-H), 7.1 (m, 5, 0-), 7.6 (d,
1, J=10 Hz, -CONH-), and ~ 9 (s, 1, -COOH).
EXAMPLE 28-31: PREPARATION OF 3-(((l-METHYL-lH-TETRAZOL-
5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4- ;~
CARBOXYLIC ACID IN FLUOROBENZENE ~-
3-(((1-Methyl-lH-tetrazol-5-yl)thio)methyl)-7-
(2-(2-thienyl)acetamido)-3-cephem-4-carboxylic acid was
prepared in four different reactions, each utilizing a
different cephalosporanic acid starting material, as follows:
28: 3-(propionyloxymethyl)-7-(2-(2-thienyl)acetamido)-
3-cephem-4-carboxylic acid (41 mg.; 0.1 mmole)
29: 3-(2-methylpropionyloxymethyl~ 7-(2-(2-thienyl)-
acetamido)-3-cephem-4-carboxylic acid (42 mg.;
0.1 mmole) ~
30: 3-(n-butyryloxymethyl)-7-(2-(2-thienyl)acetamido)- -
3-cephem-4-carboxylic acid (42 mg.; 0.1 mmole) -
31: 3-(cyclobutylcarbonyloxymethyl)-7-(2-(2-thienyl)-
acetamido)-3-cephem-4-carboxylic acid (43 mg.;
0.1 mmole)
In each instance the compound was suspended in
10 ml. of fluorobenzene (which had been dried over an
aluminosilicate desiccant sold under the term 4-A Linde
- molecular sieve). l-Methyl-lH-tetrazole-5-thiol (in each
reaction 18 mg.; 0.15 mmole) was added and the reaction
mixture was refluxed for about 24 hours. In each reaction
the product precipitated and was separated by filtration and
dried. TLC of the product in ethyl acetate/acetic acid
showed that each displacement had been quantitative. The
X-4902A -54-
. ~ . ;. . - '
. .
. . . .. ; . . .
L49~
... .. .. . ................. .... .
NMR of each product was consistent with that of an authentic :.
sample of the product prepared by aqueous displacement.
EXA~P~E 32: PREPARATION OF 3-(((3-METHYL-1,2,4-OXADIAZOL- -
5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
~ .. ~... , ., .. ,.. ,... ,,.. , ,, ,,,, ,, , ,,,, ~ .
CARBOXYLIC ACID IN 1,1,2-TRICHLOROETHANE :
7-(2-(2-Thienyl)acetamido)cephalosporanic acid .
(396 mg.; 1 mmole) was suspended in 40 ml. of 1,1,2-
trichloroethane and 3-methyl-1,2,4-oxadiazole-5-thiol (116 i
mg.; 1 mmole) added. The reaction mixture was heated to
. .... .. ~10.. 113C. in.an oil.bath.for!about.. 3 hours.,.then allowed to........ .-:
cool overnight and evaporated to an oil. Ethyl acetate and -
saturated sodium bicarbonate were added. The ethyl acetate
portion was rinsed again with saturated sodium bicarbonate,
and the aqueous portions were combined and extracted again
with ethyl acetate. The aqueous portion was layered with ~::
~ ~ -- - . - . ........................ . . . ,. : . ,
fresh ethyl acetate, cooled in an ice bath, and the pH :
adjusted to 2.5 with 20 percent HCl. The layers were
separated and the aqueous layer extracted again. The ethyl ..
acetate layer was washed, combined with saturated sodium
20 chloride, dried over magnesium sulfate, filtered, and ~
....... ~ . evaporated. .1,1,2-Trichloroethane was added to the residue, . . .: .
whereupon the product formed a solid, 220 mg. (48 percent
yield). Identity of ~he product was confirmed by NMR, IR,
UV, and bioautogram. NMR (DMSO-d6) ~ 2.35 (s, 3, CH3 on
xadiazole.).,.3-.7...(q,. 2, J-18 Hz, 2-CH2), 3.8...(s., 2,.-CH2C~N~
.. 4.4 (q, 2, J=14 Hz, 3 CH2S-), 5.1 (d, 1, J=5 Hz, C6-H),
5.6 (q, 1, J=4 Hz, C~-H)~ 6.9, 7.3 (t, d, 3, thiophene H),
and 9.1 (d, 1, J=8 Hz, -CH2CONH-).
.. ., ~ .. .... .... .... .. . . . . . ..... . .. .. . .. ............. . . .. ... .. ..... .. . .. . ..... . ..... . ....
...... . . .. . . . .. . .
X-4902A -55- .
: ., : - ', .',.. :. , ' .
.. .. . . . ..
. .
EXAMPLE 33: PREPARATION OF 3~ lH-1,3,4-TRIAZOL-5-YL)- -
THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CAR-
BOXYLIC ACID IN 1,1,2-TRICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(396 mg.; 1 mmole) was suspended in 30 ml. of 1,1,2-
trichloroethane and lH-1,3,4-triazole-5-thiol (100 mg.S 1
mmole) added. The reaction mixture was heated to 105~C.
Within 30 minutes, product was precipitating. Heating to
100C. was continued for about 6 hour~, at which time the
reaction mixture was cooled to room temperature. Product
was collected by filtration and rinsed with 1,1,2-tri- - ;
chloroethane 380 mg. (87 percent yield). The identity of
the product was confirmed by IR, NMR, UV, and bioautogram.
Bioautogram also showed the presence of the cephalosporanic
acid s~arting material, which high pre~sure liquid chroma-
tography showed to be pre~ent in the amount of 6.8 percent.
NMR (DMSO-d6) ~ 3.7 (s, 2, 2-CH2), 3.8 (s, 2, -CH2CONH~
4.2 (q, 2, J=5 Hz, 3-CH2S-), 5.1 (d, 1, J=5 Hz, C6-H), 5.7
(q, 1, J=4 Hz, C7-H), 7.0, 7.4 (t, d, 3, thiophene _), 8.45
20 (s, 3, triazole \NH), and 9.13 (d, 1, J=8 Hz, -C~2CON~I-).
EXAMPLE 34: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-PHENYLACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID IN CHLOROFORM -
7-(2-Phenylacetamido)cephalosporanic acid (195 -
mg.; 0.5 mmole) was suspended in 75 ml. of ahloroform and 1-
methyl-lH-tetrazole-5-thiol (75 mg.; 0.65 mmole) added. The
reaction mixture wa~ heated in an oil bath at 80-85C. for
3Q
X-4902A -56-
: . . , , ,, . ; . . ,, : ,
, . , , :, .
3~
3 hours and 60 ml. distilled off. TLC showed very little ~ ~ -
product, whereupon 20 ml. of 1,2-dichloroethane was added
and heating continued overnight. TLC of the reaction
mixture the following morning showed little cephalosporanic
acid starting material. After a total reaction time of
26 hours, the reaction mixture was cooled to room tempera-
ture. The product was worked up in essentially the same
procedures as reported in Example 32, yielding 80 mg. of
puriPied material (35 percent yield). Identity of the
product was confirmed by NMR, IR, UV, and bioautogram. NMR
(DMSO-d6) ~ 3.6 (s, 2, 2-CH2), 3.7 (s, 2, -CH2CONH-)i 4.0
(s, 3, C_3 on tetrazole), 4.3 (s, 2, 3-CH2S-~, 5.1 (d, 1,
J=5 Hz, C6-H), 5.7 (q, 1, J=4 Hz, C7-_), 7.3 (s, S, 0), and
9.13 (s, 1, J=8 Hz, -CH2CONH-).
EXAMPLE 35: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-ACETAMIDO-3-CEPHEM-4-CARBOXYLIC ACID IN
1,2-DICHLOROETHANE
7-Acetamidocephalosporanic acid (314 mg.; 1
mmole) and l-methyl-lH-tetrazole-5-thiol (98 mg.; 0.65
20 mmole) were mixed in 70 ml. oP 1,2-dichloroethane; 50 ml.
were distilled off and the reaction mixture than refluxed
for about 24 hours.
The reaction mixture was worked up in essentially
the procedures reported in Example 32 yielding 120 mg. oP
product (yield, 32 percent). Identity oP the product was
confirmed by NMR, IR, W , and bioautogram. NMR (DMSO-d6) ~
1.97 (s, 3, OEI3CONH-), 3.7 (s, 2, 2-CH2), 4.0 (s, 3, CH3 on
tetrazole), 4.35 (s, 2, 3-CH2S-), 5.1 (d, 1, J=5 Hz, C6H), ;
5.7 (q, 1, J=4 Hz, C7-_), and 8.8 (d, 1, J-8 Hz, -CH2CONH).
-
X-4902A -57-
''' ''." '' .',; , .";, ,", ,',', ', ', ,, ' ' " ' ' '', '' "'', "":' ~ ' ', ., ' ' ' ' ' ., , ' ' ',. '' ' '
; ' ' :: '
: '
133~
EXAMPLE 36: PREPAR~TION OF 3-(((3-METHYL-1,2,4-THIADIAZOL-
5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-( 2-(2-Thienyl)acetamido)cephalosporanic acid ~-
(200 mg.; 0.5 mmole) and 3-methyl-1,2,4-thiadiazole-5-
thiol (85 mg.; 0.65 mmole) were mixed in 50 ml. of 1,2-
dichloroethane and heated to 95C. on an oil bath. The bath
temperature was maintained at 90C. for 19 hours. The
reaction mixture was then removed from the bath and allowed
to cool and refrigerated for a day. 1'hree volumes of ethyl
acetate were then added and washed with two 50 ml. portions
of saturated sodium bicarbonate. The combined aqueous
portions were extracted with ethyl acetate, layered with
fresh ethyl acetate, and cooled in an ice bath. The pH was
adjusted to 2Ø The layers were separated and the aqueous
layer extracted again with ethyl acet~te. The combined ~-
ethyl acetate portions were washed with saturated sodium
chloride, dried over magnesium sulfate, filtered, and
evaporated, yielding 252 mg. of a foam. It crystallized
20 from acetone/diethyl ether, 153 mg. (65 percent yield).
Identity of the product was confirmed by NMR, UV, IR, and
bioautogram. NMR ~DMSO-d6) ~ 3.7 (s, 2, 2-CH2), 3.8 (s,
2, -C _2CONH), 4.5 (q, 2, J=15 Hz, 3-C~ S-), 5.1 ~d, 1, J=5
Hæ, C6-H), 5.7 (q, 1, J=4 Hz, C7-H), 6.96, 7.38 (t, d, 3, ~
thiophene _), and 9.13 (d, 1, J=8 Hz, -CH2CONH). ;
...
''.
: .. '
~ ~-
X-4902A -58-
.: . :,- . . - . : ~ ,' ': ' '
,- , , , :
,, , : ,, , . :
~O~l3~49~
EXAMPLE 37: PREPARATION OF 3-(((2-PYRIMIDINYL)THIO)METHYL)-
7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN
ACETONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; S mmole) and 2-mercaptopyrimidine (0.62 grams;
5.5 mmole) were mixed in 25 ml. of dry acetonitrile and the
mixture refluxed overnight (16 hours) with stirring. TLC
showed complete conversion to product. The reaction mixture L
was cooled to room temperature and the product separated by
10 filtration. It was then washed with 50 ml. of acetonitrile
and vacuum dried at 50C. for 4 hours, 1.86 gram~ of off-
white crystals (83.0 percent yield), m.p. 217C. (dec). NMR
in DMSO-d6 showed neither starting material. The identity
of the product was confirmed by IR, UV, and NMR. NMR
(DMSO6-d6) ~ 3.55 (q, 2, 2-CH2, J=18 Hz), 3.74 (s, 2,
-C_2CONH), 4.28 (q, 2, 3-CH2S-, J=13 Hz), 5.00 (d, 1, C6-H,
J=5 Hz~, 5.64 (q, 1, C7-H, J=5 Hz, J=9 Hz), 7.08 (m) and
8.52 (d) (6, thiophene and pyrimidine H~, and 9.00 (d,
1, -CH2CONH-, J=9 Hz).
20 EXAMPLE 38: PREPARATION OF 3-(((2-PYRIMIDINYL)THIO)METHYL)-
7-(2-(2-THIENYL)ACETAMIDO)-3-CE~EM-4-CARBOXYLIC ACID IN
ACETIC ACID
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole), 2-mercaptopyrimidine (0.6 gram; 5.4
mmole), and sodium acetate (0.41 gram; 5 mmole) in 25 ml. of
glacial acetic acid were heated ko 85C. for 4 hours.
Product crystallized during the course of reaction. The
reaction mixture was then cooled to room temperature and the
product separated by filtration, washed in aceti~ acid, and `
X-4902A -59-
: ~:
:-
.. , , , '.
~ f~
dried, 1.58 grams (70.5 percent yield) of white crystals,m.p. 218C. (dec.). Identity of the product was confirmed
by IR, UV, NMR, and elemental analysis. The NMR was identical
with the NMR of the product of Example 37.
EXAMPLE 39: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)CEPHALO~PORANIC
ACID IN ACETIC ACID
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole), 1-methyl-lH-tetrazole-5-thiol (0.81 ~ -
gram; 7 mmole), and sodium acetate (0.41 gram; 5 mmole) in
25 ml. of glacial acetic acid were heated to 75-77C. and
held there over 8 hours. T~C was run every hour and at
eight hours, TLC showed that the reaction was complete. The
reaction mixture was cooled to 40-45C. and the acetic acid
removed under vacuum. To the residue was then added 50 ml.
of ethyl acetate and 50 ml. of water. The aqueous layer was
acidified to pH 1.5 with lN H2SO4. The ethyl acetate layer
was separated, dried over anhydrous sodium sulfate, and the
ethyl acetate removed on a rotary evaporator. The residue,
a light weight oil, was redissolved in 50 ml. of ethanol and
a solution of 2 ml. of dicyclohexylamine (10.2 mmole) in
10 ml. of ethanol was added. The product precipi-tated
almost immediately as the dicyclohexylamine salt but was
stirred an additional 15 minutes; it was then separated by
filtration, washed with ethanol and dried, 1.2 grams (37.8
percent yield), m.p. 185-6C. IR, NMR, and UV were identical
with previously prepared samples of the same product.
''' ' '
'.
X-4902A -60-
. .
.
33~
EXAMPLE 40: PREPARATION OF 3-(((1-BE~ZYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-benzyl-lH-tetrazole-5-thiol (1.44
grams; 7.5 mmole) were added to 25 ml. of 1,2-dichloroethane
and the reaction mixture heated to 85C. in an oil bath and
maintained at that temperature overnight, with stirring. -
TLC showed no cephalosporanic acid starting material.
Solvent was removed on a rotary evaporator, leaving a foam.
To the foam, 25 ml. of methanol was added and the mixture
warmed on a steam bath until the sample dissolved. Solvent
was evaporated on a rotary evaporator and the product crys-
tallized. It was stirred for 15 minutes, then separated by
filtration, washed with methanol, and vacuum dried, 1.6
grams (60.6 percent yield), m.p. 171-171.5C.
EXAMPLE 41: PREPAR~TION OF 3-AMIDINOTHIOMETHYL-7-(2-(2-
, . ... . .
~ THIENYL)ACETAMI~O)-3-CEPHEM-4-CARBOXYLIC ACID IN ACETONITRILE
- :
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(3.12 grams; 8 mmole) and ~hiourea (912 mg.; 12 mmole) in
15 ml. of acetonitrile ~which had been dried over an alumi-
nosilicate desiccant sold under the term 4-A Linde molecular
sieve) were heated at 87C. with stirring for 24 hours.
Within one hour, precipitate began ~orming in the reaction
mixture.
At the end of 24 hours, the product was separated
by filtration of the hot reaction mixture, and dried, 2.5
grams (76 percent yield). Elemental analysis showed
.
-~
X-4902A ~61-
,
.. .. .. .. . .. .
3~
Theory: C, 43 68i H, 3.91; N, 13.58.
Found: C, 43.50; H, 4.03; N, 13.29.
NMR confirmed the identity of the product.
EXAMPLE 42: PREPARATION OF 3-BENZOYLTHIOMETHYL-7-(2-~2-
THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN ISOPROPYL
ACETATE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(1.0 gram; 2.5 mmole) and thiobenæoic acid (510 mg.; 3.7
mmole) were suspended in 20 ml. of anhydrous isopropyl
acetate and the suspension heated under reflux for 31 hours,
by which time all reactants were in solution. Solvent was
evaporated under vacuumr leaving a residue which TLC indicated
was a 60-40 mixture of product and unreacted starting
material.
Preparative thick-layer chromatography over silica -~
plates using acetone: acetic acid (16:1) as eluent afforded
a purified sample of product. NMR (DMSO-d6/D2O) ~ 3.50 - ;
(ABq, 2H, 2-CH2, J=6 Hz, J=l9 Hz), 3.82 (s, 2H, -CH2CONH-),
4.20 (d, 2H, 3-CH2S-, J=5 Hz), 5.03 (d, lH, C6-_, J=5 Hz),
20 5.69 (d, lH, C7-H, J=5 Hz), 6.89-8.0 (m, 8H, aromatic H).
EXAMPLE 43: PREeA~ATION OF 3-((PHENYLTHIO)METHYL)-7-(2-
(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN 1,2-
DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and benzenethiol (0.75 ml.; 7.5 mmole)
were added to 25 ml. of 1,2-dichloroethane. The reaction
mixture was refluxed overnight; TLC showed no cephalosporanic
acid starting material remaining.
,
X-4902A -62-
-` ~L0~3~4~
Solvent was evaporated on a rotary evaporator and
25 ml. of ethanol added to the residue and heated to get the
residue into partial solution. To get the remainder into
solution, 25 ml. of methanol was added and heated. The
resulting solution was filtered through cotton, treated with
2.0 grams of carbon, stirred 5 minutes, and filtered through
filter aid. Solvent was removed on a rotary evaporator.
NMR of the residue showed some thiol starting material
present.
The residue was slurried in 25 ml. of isopropyl
acetate, filtered, and refrigerated overnight. The filtrate
was then diluted with 25 ml. of ethyl acetate and washed
with 25 ml. of dilute sodium bicarbonate to remove thiol~
The ethyl acetate layer was added to 25 ml. o~ water and the
.
pH at 8.5 was adjasted to pH 1.4 with H2SO4. The ethyl
acetate layer was then dried over magnesium sulfate,
,
filtered, and evaporated to a foam on a rotary evaporator,
0.92 gram (19.4 percent yield). NMR (DMSO-d6) ~ 3.58 (m, 2~ -
2-CH2), 3.75 (s, 2, -CH2CONH-), 4.12 (q, 2, 3-CH2S-, J=13 ~ -
Hz), 5-08 (d, 1, C6-_, J=5 Hz), 5.66 (q, 1, C7-_, J=5 Hz,
J=9 Hz), 7.15 (m, 8, thiophene H and phenyl _), and 9.10 (d,
CH2CONFf-~ J=9 Hz).
EXAMPLE 44: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
~ 2-YL)THIO)METHYL)-7-(2-PHENYLACETAMIDO)-3-CEPHEM-4-CAR-
- BOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-Phenylacetamido)cephalosporanic acid (1.95
grams; 5 mmole) and 5-methyl-1,3,4-thiadiazole-2-thiol (0.99
gram; 7.5 mmole) in 25 ml. of 1,2-dichloroethane were
refluxed for 16 hours. TLC showed a trace of cephalosporanic
X-4902A -63-
., ~ . .~ . . : : .
- ~ ~133~
acid starting material. Product crystallized during the
reaction.
The reaction mixture was cooled to 0-5C. and
filtered, and the product washed with cold 1,2-dichloro-
ethane and vacuum dried, 1.82 grams (78.8 percent yield),
m.p. 171-2C. NMR (DMSO-d6) ~ 2.70 (s, 3, -CH3 of thia-
diazolyl), 3.58 (s, 2, -C_2CONH-), 3.70 (broad s, 2, 2-CH2),
4.~0 (q, 2, 3-C_2S-, J=13 Hz), 5.12 (d, 1, C6-H, J=5 Hz), ~ ~;
5.72 (q, 1, C7-_, J=5 Hz, J=9 Hz), 7.28 (s, 5, phenyl H),
10 and 9.08 (d, 1, -CH2CON_-, J=9 Hz).
EXAMPLE 45: PREPARATION OF 3-((METHYLTHIO)METHYL)-7-(2-
(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN METHYLENE
CHLORIDE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid (24
grams; 60 mmole), 7.0 ml. of methanethiol, and 600 ml. of
methylene chloride were stirred and heated in a bomb for 18
hours at 84-86C. The reaction mixture was then cooled to
room temperature. A small amount of insoluble material was
filtered off and the filtrate was evaporated. The residue
was taken up in ethyl acetate and filtered. The fil-trate
was layered with about 150 ml. of water and stirred; lN NaOH
was added dropwise to a pH 5.5. The aqueous phase was
separated and concentrated to a volume of about 75 ml., then
diluted to 700 ml. with water, and glacial acetic acid added
to pH 3.8. An amorphous solid precipi~ated and was stirred
for 3 hours on an i~e bath, then refrigerated overnight.
The solid was then filtered. The filtrate was
layered with 100 ml. of ethyl acetate ~nd the pH adjusted to
2.0 with concentrated HC1. The organic phase was separated
;~
X-4902A -64-
', :~: '
- ,
,. .
.
3~4~
and extracted with one-100 ml. portion and one-150 ml.
portion of ethyl acetate. The combined organic phases were
dried over magnesium sulfate, then stripped of solvent,
yielding an amber colored foam. It was dissolved in 50 ml.
of ethyl acetate, seeded with the intended product, and held
in a refrigerator overnight. Crystals had formed; they were
separated by filtration, rinsed with cold isopropyl acetate,
and vacuum dried at 50C., 3.2 grams (14 percent yield).
NMR confirmed the identity of the product, ~ 2.00 (s, 3,
3-CH2SCH3)~ 3-74 (m, 3-CH2S-, 2-CH2, and -CH2CONH-)~ 5-14
(d, 1, C6-H, J=5 Hz), 6.64 (q, 1, C7-H, J=5 Hz, J=9 Hz), ;
7.15 (m, 3, thiophene H), and 9.12 (d, 1, -CH2CONH-, J=9
Hz). ~
EXAMPLE 46: PREPARATION OF 7-(2-FORMYLOXY-2-PHENYLACET- -
AMIDO)-3-(1-METHYL-lH-TETRAZOL-5-YL)THIO)METHYL)-3-CEPHEM-
4-CARBOXYLIC ACID SODIUM SALT IN BENZENE
7-(2-Formyloxy-2-phenylacetamido)cephalosporanic
acid (2.17 grams; 4.65 mmoles) and l-methyl-lH-tetrazole-
5-thiol (0.87 gram; 7.5 mmole) in 25 ml. of benzene were
rèfluxed (about 80C.) for 12 hours. A layer of soft
plastic deposited on the glass vessel during the reaction.
On cooling, the plastic hardened to a glassy solid. TLC
showed that the solution contained only a trace of product
and that the plastic was mostly product with some deformylated
product. The reaction mixture was evaporated with added
acetone to a foam which was dissolved in 35 ml. of dry
acetone and treated with 10 ml. of a solution of ]..25 grams
of sodium 2-ethylhexanoate (7.5 mmole) in acetone. The
product crystallized as the sodium salt. After an hour, the
X-4902A -65-
.
~ . . , . ~
83~
product was filtered, washed with 20 ml. of acetone anddried, 1.38 grams (58 percent yield). The identity of the
product was confirmed by TLC and NMR. NMR (D2O) ~ 3.48 (q,
2, 2-CH2, J-18 Hz), 4.00 (s, 3, -CH3 of tetrazolyl), 4.10
(m, 2, 3-CH2S-), 5.05 (d, 1, C6-H, J=5 Hz), 5.70 (d, 1, C7-
H, J-5 Hz), 6~24 (s, 1, -CHCONH-), 7.50 (m, 5, phenyl H),
O
and 8.33 (s, 1, -OCH).
EXAMPLE 47: PREPARATION OF 7-(2-FORMYLOXY-2-PHENYLACET-
AMIDO)-3-(((1-METHYL-lH-TETRAZOL-l-YL)THIO)METHYL)-3-
CEPHEM-4-CARBOXYLIC ACID IN CARBON TETRACHLORIDE
7-(2-Formyloxy-2-phenylacetamido)cephalosporanic
acLd (2.17 grams; 4.65 mmole) and 1-methyl-lH-tetrazole-5-
thiol (0.87 gram; 7.5 mmoles) in 25 ml. of carbon tetra-
chloride were refluxed (about 77C.) for 12 hours. The
reaction mixture was then cooled to room temperature and the
supernatant decanted from a hardened semi-solid. To the
solid was added 25 ml. of 1,2-dichloroethane, with warming.
The product crystallized. The mixture was cooled to room
temperature and stirred for about an hour, then the product
was separated by filtration, washed with 10 ml. of 1,2-
dichloroethane, and vacuum dried overnight at 45C., 1.54
grams of white crystals (65 percent yield). Identity of the
product was confirmed by NMR and TLC, the latter of which
showed a small trace of deformylated product. The NMR was
identical with the NMR of the product of Example 16.
- - -
X-4902A -66-
,, ....,
,
. .
, - . : .. . : . . - .
-, : . , . ,~ . :
~33~
PREPARATION 48: PREPARATION OF 3-METHYL-1,2,4-OXADIAZOLE
5-THIOL
Acetamide oxime (30 grams; 0.4 mole~, carbon
disulfide (100 ml.; 1.66 mole) and triethylamine (56 ml.;
0.4 mole) were mixed and stirred in l liter of pyridine. A
stream of nitrogen gas was bubbled through a carbon disulfide ;
solution and then passed over the reaction mixture. The
mixture was heated in an oil bath at 70C. for three days,
then evaporated to an oil, to which ethyl acetate and
saturated sodium carbonate were added. The layers were
separated and the organic layer was again washed with saturated
sodium bicarbonate. The combined sodium bicarbonate washes
were extracted with ethyl acetate and the ethyl acetate
extracts were discarded. The aqueous portion was layered
with fresh ethyl acetate and cooled in an ice bath. The pH
was adjusted to 2.5 with 20 percent HCl and sodium chloride
was added to saturate the solution. It was then extracted
with ethyl acetate, rinsed with saturated sodium chloride
solution, and dried over anhydrous magnesium sulfate. The
solution was filtered and evaporated to one-half its original
volume. An equal volume of carbon tetrachloride was added
and evaporation was continued until the product crystallized,
24.8 grams (52 percent yield).
EXAMPLE 49: PREPARATION OF 3-(((3-METHYL-1,2,4-OXADIAZOL-
; 5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)-3-(carbamoyloxymethyl)-
3-cephem-4-carboxylic acid (100 mg.; 0.25 mmole) was suspended
in 25 ml. of 1,2-dichloroethane and stirred while 3-methyl-
X-4902A -67-
" . . .
., . ~ . . . .
, '
~83~
1,2,4-oxadiazole-5-thiol (35 mg.; 0.30 mmole) was added.
The reaction mixture was heated on an oil bath at 110C., to
distill off 5 ml. of solvent including any trace amount of
water. The oil bath temperature was lowered to 90-95C. and
khe reaction mixture maintained for 19 hours, then cooled to
room temperature. Insoluble material was removed by fil-
tration and rinsed with 1,2-dichloroethane and diethyl
ether. It was determined by TLC to be unreacted starting
material with only a trace amount of product.
The filtrate was evaporated and the residual oil
partitioned between sodium bicarbonate ~olution and ethyl
acetate. The aqueous solution was layered with fresh ethyl
acetate, cooled in an ice bath, and the pH adjusted to 2.5
with 20 percent HC1. The organic layer was removed. The
aqueous layer was extracted with fresh ethyl acetate and the
combined ethyl acetate solutions were washed with saturated
NaCl solution, dried over magnesium sulfatel filtered, and
evaporated to an oil. Crystallization from a 1:1 solution
containing hexane and diethyl ether gave 13 mg. of product,
(11.5 percent yield). The identity of the product was
confirmed by NMR and IR, which were identical with the same
compound prepared as reported in Example 32.
EXAMPLE 50: PREPARATION OF 3-(((l-METHYL-lH-TETRAæOL-S-
YL)THIO)METHYL)-7-(2-(tert-BUTOXYCARBONYL)-2-PHENYLACET~
AMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(tert-Butoxycarbonyl)-2-phenylacetamido)-
cephalosporanic acid (490 mg.; 1 mmole) and l-methyl-lH-
tetrazole-5-thiol (145 mg.; 1.25 mmole) in 15 ml. of dry
nitromethane were heated at 85-90C. ~or 8 hours under
X-4902A -63
.'. ;' '.
:''. :.
: ' : :.... . ': ' ' .
, '- , ' ' . , . , '', , :
4~.
nitrogen. TLC showed product, excess thiol reactant, and
decarboxylated product, but no cephalosporanic acid starting
material. The nitromethane was removed by evaporation. The
residual orange foam was dissolved in 10 ml. of saturated
sodium bicarbonate, 20 ml. of water was added, and the
mixture was washed successively with ethyl aceta~e until the
washes were clear. l'he ethyl acetate washes were combined,
20 ml. of water was added, and the mixture cooled to 0C.
and adjusted to pH 2.2 with 20 percent HCl. The layers were
separated and the aqueous layer washed with ethyl acetate.
The ethyl acetate portions were combined, washed with
saturated sodium chloride, dried over magnesium sulfate,
filtered, and evaporated to a brown foam, 455 mg. (83 percent
yield). TLC was excellent, showing the presence of product,
a trace of thiol, and a trace of decarboxylated product.
The 455 mg. of product (0.833 mmole) were dissolved
in 7 ml. of ethyl acetate and lithium acetate (0.833 ml.)
was added dropwise with stirring, resulting in a brown
precipitate, the lithium salt. It was separated by fil-
tration, washed with ethyl acetate, and dried overnightunder vacuum a~ room temperature, 368 mg. (80 percent yield).
The identity of the product was confirmed by TLC, NMR,
bioautogram, elemental analysis, IR, and UV. TLC showed a
trace of decarboxylated material. NMR (CDC13) ~ 1.4 (s,
9, -COOtertC4Hg), 3.6 (s, 2, 2-CH2), 3.85 (s, 3, CH3 of
tetrazole), 4.3 (s, 2, 3-CH2S-), 4.44 and 4.45 (2s, 1,
0CH-), 4.9 (d, 1, J=6 Hz, C6-H), 5.8 (q, 1, J=6 Hz, C7-H),
7.35 (s, 5, 0), 8.2 and 7.8 (2d, 1, J~9 Hz, -CONH-), and 9.3
(s, 1, -COOH). ~
-
X-4902A -69-
. . ~ . - ~ , . : :
... : .. . .. .. . . . .
:
EXAMPLE 51: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(N-((1,3-DIMETHYLUREIDO)CARBONYL)-2-
PHENYLGLYCYLAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN NITROMETIIANE
7-(N-((1,3-Dimethylureido)carbonyl)-2-phenyl- -
glycylamido)cephalosporanic acid (130 mg.; 0.25 mmole) was
suspended in 5 ml. of nitromethane and l-methyl-lH-tetra-
zole-5-thiol (43.5 mg.; 0.375 mmole) was added. The reaction
mixture was heated at 85 under nitrogen for 12 hours, then
held at room temperature over a weekendO The reaction
mixture was then filtered and the solid washed with a small
amount o~ nitromethane and dried in a vacuum oven at 35,
83 mg. (58 percent yield). The identity of the product was
confirmed by TLC, NMR, IR, UV, elemental analysis and
bioautogram. NMR (DMSO-d6) ~ 2.65 (d, 3, J=4 Hz, -CONHC_3),
3.15 (s, 3, CONC_3CO-), 3.6 (s, 2, 2-CH2), 3.9 (s, 3, CH3 on
tetrazole), 4.3 (s, 2, 3-CH2S-), 5.0 (d, 1, J=5 Hz, C6-H),
5.5 (d, 1, J=7 Hz, 0CHCO ), 5.7 (q, 1, C7-H), 5.8 (q, 1,
N-
-CON_CH3), 7.4 (s, 5, 0), 9.3 (d, 1, J=8 Hz, 0CHCONH-),
and 10 (d, 1, J=8 Hz, 0CH-CO-).
N_
EXAMPLE 52: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-YL)- -
THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID IN ACETONITRILE, TETRABUTYLAMMONIUM IODIDE ADDED
7-(2-(2-Thienyl)acetamido)cephalosporanic acid (2.0
grams; S mmole)j l-methyl-lH-tetrazole-5-thiol (1.2 grams;
10 mmole), and tetrabutylammonium iodide (0.2 gram) were ~ -
- mixed in 25 ml. of dry acetonitrile, heated to reflux, and
refluxed for 8 hours. The reaction miY~ture was then cooled
X-4902A -70-
.
. .
.
... .
:.. . ' ' ~
.
-, . . . . .
.
3~L4~
to room temperature and solvent removed on a rotary evaporator.
The residue was treated with a hot mixture of 25 ml. of
isopropyl acetate and 5 ml. of acetonitrile, filtered, and
allowed to cool slowly. The product precipitated as light
cream colored crystals and was filtered, washed with isopropyl
acetate, and dried, 1.30 grams (57.5 percent yield). The
NMR was identical with the NMR on the product of Example 5.
EXAMPLE 53: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-~-CARBOXYLIC
ACID IN 1,2-DICHLOROETHANE (TETRABUTYLAMMONIUM IODIDE ADDED)
The reaction reported in Example 52 was repeated
except that 1,2-dichloroethane was employed as solvent.
Dicyclohexylamine (2 ml.) was added to obtain the product as
the dicyclohexylamine salt, 1.55 grams (48.9 percent yield).
The NMR was identical with the N~R on the product of Example
1. ~
EXAMPLE 54: PREPARATION OF 3-((PHENYLTHIO)METHYL)-7-
(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYI.IC ACID IN
1,2-DICHLOROETHANE, 1 METHYL-5-(METHYLTHIO)-lH-TETRAZOLE
ADDED
7-(2-(2-Thienyl)acetamido)cephalosporanic acid -
(2.0 grams; 5 mmole), benzenethiol (0.75 ml.; 7.5 mmole),
and 0.65 gram of 1-methyl-5-(me-thylthio) lH-tetrazole were
mixed in 25 ml. of 1,2-dichloroethane and heated to reflux.
The reaction was followed by TLC and appeared to be complete
in 14 hours. Solvent was removed on a rotary evaporator and
the residue was extracted repeatedly with diethyl ether.
Exhaustive removal of solvent left 1.66 grams of tan solid
product (74 percent yield). IR and NMR were identical with
the qame product prepared as described in preceding examples.
X-4902A -71
. .
EXAMPLE 55: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYLACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN ISOPROPANOL
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(2.0 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.87
gram; 7.5 mmole) in 25 ml. of isopropanol were placed in a
flask (equipped with a condenser, with a drying tube con-
taining anhydrous calcium sulfate sold under the trademar~
Drierite). The flask was immersed in an oil bath at 84-85C.
The reaction was followed by TLC. After 40 hours at 82-83C.
only half of the cephalosporanic acid had reacted.
EXAMPLE 56: PREPARATION OF 3-((2-OXAZO~YLTHIO)METHYL)-7-
(2-(lH-TETRAZOL-l-YL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID
IN NITROMETHANE
A suspension of 7-(2-(lH-tetrazol-l-yl)acetamido)-
cephalosporanic acid (0.3~ gram; 1.0 mmole) and 2-oxazole- ~ -
thiol (0.11 gram; 1.1 mmo]e) in 5 ml. of nitromethane was
immersed in an oil bath at 90-91Co An atmosphere of dry -~
nitrogen was maintained 0~7er the reaction mixture. After 20
minutes, all of the reactants had dissolved, and after 35
minutes, product began to crystallize. After 6 hours the
reaction mixture was cooled to room temperature and the
product was filtered, washed with 7 ml. of nitromethane, air
drled, and vacuum dried for 3 hours at 40C., yielding 0.36
gram (85.7 percent yield of off white crystals, m.p. 196C.
(dec). NMR showed the product to be 70 percent of the
desired product and 30 percent of the cephalosporanic acid -
starting material.
X-4902A -72-
: -. :. -. . , . : :. . . . . . .
, . . . . . . . . . .. . .
,: : , : . , .
': . . -. . ' . ,- : . ' . ' :
.... . . . . .
~3 lL~
The product was recrystallized from 5 ml. of
DMSO-d6 and 10 ml. of water, separated by filtration, washed
with 5 ml. of 2:1 water:DMSO-d6, air dried, and vacuum dried
at 50C. for 6 hours, yielding 0.26 gram. NMR showed that
the product contained 87 percent of the desired product and
13 percent of starting cephalosporanic acid. The recrystal-
lization was repeated with 3 ml. of DMSO-d6 and 6 ml. of
water, stirring for 1 hour, resulting in 0.21 gram. NMR
(DMSO-d6) indicated that the starting cephalosporanic acid
had been reduced to 5 percent, and confirmed the identity of
the desired product: ~ 3.75 (s, 2, 2-CH2), 4.34 (q, 2,
3-CH2S-, J-14 Hz), 5.16 (d, 1, C6-H, J=5 Hz), 5.76 (q, 1,
C7-H, J=5 Hz, J=9 Hz), 5.44 (s, 2, -CH2CONH-), 7.28 (s, 1,
oxazole C4-_), 8.14 (s, 1, oxazole C5-H), 9.37 (s, 1,
tetrazole _), and 9.53 (d, 1, -CONH-, J=9 Hz).
EXAMPLE 57: PREPARATION OF 3-((2-OXAZOLYLTHIO)METHYL)-
7-(2-FORMYLOXY-2-PHENYLACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID
.
IN 1,2-DICHLOROETHANE
A solution of 7-(2-formyloxy-2-phenylacetamido)-
cephalosporanic acid methylene chloride solvate (0.52 gram;
1 mmole) and 2-oxazolethiole (0.12 gram; 1.1 mmole) in
20 ml. of 1,2-dichloroethane was refluxed for about 16 hours
and then cooled to room temperature. TLC showed clear con-
version to product. The reaction mixture was concentrated
on a rotary evaporator to 10 ml.; and upon standing, some-
what gelatinous crystals formed. They were filtered, washed
with 1,2-dichloroethane, and dried, yielding 0.12 gram of
gray solid.
X-4902A -73-
.- - ~ .. . . .
.. ..
,, :
3~4~L
Exhaustive removal of solvent from filtrate left
0.45 gram of light yellow foam. It was triturated with
25 ml. of diethyl ether, filtered, washed with diethyl
ether, and dried yielding 0.21 gram of light yellow powder.
The identity of the product was confirmed by NMR, ~ 3.56 (m,
2, 2-CH2), 4.24 (~, 2, 3-CH2S-), J=13 Hz), 5.00 (d, 1,
C -H, J=5 Hz), 5.70 (q, 1, C -H, J=5 Hz~ J=9 Hz), 6.14 (s,
6 - - 7 - -
1, e~ ~a--cH-), 7.25 (s, 1, oxazole C4-H), 7.45 (m, 5, phenyl
H), 8.12 (s, 1, oxazole C5-H), 8.35 (s, 1, -CHO), and 9.40
(d, 1, -CONH-, J=9 Hz).
EXAMPLE 58: PREPARATION OF 3-(((4-PH~NYL-2-THIAZOLYL)-
THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-CAR-
BOXYLIC ACID IN ACETONITRILE
A mixture of 7-(2-(2-thienyl)acetamido)cepha-
losporanic acid (2.0 grams; 5 mmole) and 4-phenyl-2-
thiazolethiol (1.44 grams; 7.5 mmole) in 35 ml. of acetonitrile
was reflu~ed for 16 hours, protected from atmospheric
. . .
moisture with a drying tube containing anhydrous calcium
sulfate sold under the trademark Drierite. TLC showed clean
conversion to a new spot.
Upon cooling the reaction mixture to room tem-
perature and stirring it for 2 hours, the product crys-
tallized and was filtered, washed with acetonitrile, and
dried, 2.25 gram (85.6 percent yield, m.p. 180C. (dec.). -
NMR confirmed the identity of the product.
.
X-4902A -74-
~, ,'.',. '" '" '.;, ' ,' .' ., ''' " ., ,'.. '" ;'' " :.' ' ' ':.'"' " ,'~ ; ', ',
.: .. : ... .
~3~
EXAMPLE 59: PREPARATION OF 3-~((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-1-YL)ACETAMIDO~-3-
CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (1 92 grams; 5 mmole) and 5 methyl-1,3,4-thiadiazole-
2-thiol (0.79 gram; 6 mmole) were added to 25 ml. of alumina-
treated nitromethane and the reaction mixture heated to
95C. in an oil bath and allowecl to stir for a total of 4
hours. TLC showed conversion except for a trace of cepha-
losporanic acid starting material (less than 2 percent).
The reaction mixture was cooled to room temperature filtered,
washed with nitromethane, and vacuum dried, 2.11 grams (92.5
percent yield), m.p. 183.5C. (dec.). NMR confirmed the
identity of the product.
EXAMPLE 60: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-l-YL)ACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN PROPIONITRILE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (1.92 grams; S mmole) and 5-methyl-1,3,4-thiadiazole-
20 2-thiol (0.99 grams; 7.5 mmole) were added to 25 ml. of
propionitrile (treated over neutral alumina) and heated to
reflux (97C.). The reaction mixture was refluxed with
stirring for 9 hours. TLC showed only a trace of cepha-
losporanic acid starting material. The reaction mixture was
cooled to room temperature, filtered, washed with propionitrile,
and vacuum dried, 2.04 grams (89.5 percent yield), m.p.
186.5C. (dec.). NMR confirmed the identity of the product.
,
X-4902A -75~
., ~. .
. . ::, .. .
~33~
EXAMPLE 61: PREPARATION OF 3-(((l~METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-FORMYLOXY-2-PHENYLACETAMIDO)-3-CEPHEM-
4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-ACA (7-aminocephalosporanic acid) (13.62 grams;
0.05 mole) was suspended in 100 ml. of 1,2-dichloroethane
and trimethylsilyl acetamide (26.25 grams; 0.200 mole) was
added. The reaction mixture was heated to 40C. and the
solid dissolved to yield a cloudy solution, which was cooled
to 20C. -
A solution of 2-formyloxy-2-phenylacetyl chloride ;
(10.92 grams; 0.055 mole) in 25 ml. of 1,2-dichloroethane
was added dropwise over 20 minutes, with a temperature rise -
to 30C. The reaction mixture was stirred for 2 hours,
100 ml. o~ 1,2-dichloroethane were added, and the reaction
mixture was washed with three 100-ml. portions of water.
The water washes were combined and extracted with two ~
50-ml. portions of 1,2-dichloroethane, which were back !.:'",.'' .'
washed with 40 ml. of water. The 1,2-dichloroethane layers
were combined, stirred 20 minutes with 2.0 grams of activated
carbon sold under the trademark Darco G-60, and filtered
through a diatomaceous earth sold under the trademark Hyflo.
Total volume was 375 ml. of solution containing the desired
7-(2-formyloxy-2-phenylacetamido)cephalosporanic acid.
This solution was evaporated at 30C. to 336
grams, estimated to be 250 ml. of 1,2-dichloroethane and 22
grams of the 7-(2-formyloxy-2-phenylacetamido)cephalosporanic
acid. l-Methyl-lH-tetrazole-5-thiol (6.39 grams; 55 mmole)
in 250 ml. of 1,2-dichlorothane was added and the reaction
mixture heated to reflux. Water ~apparently remaining from
X 4902A -76-
.. : :,, .:. .. . . .
833L4~
earlier washes) was azeotroped into the condenser and beganreturning into the vessel; a trap was employed to intercept
water. TLC after 12 hours of reflux showed near normal
reaction. The reaction mixture was cooled to room tem-
perature and seeded to precipitate the product, 3-(((1-
methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-formyloxy)-
2-phenylacetamido)-3-cephem-4-carboxylic acid. After two hours,
the product was filtered and washed with 63 ml. of 1,2-
dichloroethane, 13.90 grams (56.7 percent yield based on
7-ACA). TLC of the product was clean. The NMR confirmed
the identity of the product and was identical with the NMR
of the product prepared as reported in Example 16.
EXAMPLE 62: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
~L)THIO)METHYL)-7-(2-FORMYLOXY-2-PHENYLACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-Pormyloxy-2-phenylacetamido)cephalosporanic
acid (2.33 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-
thiol (0.64 gram; 5.5 mmole) were added to 25 ml. of 1,2-
dichloroethane and the mixture refluxed for 12 hours. The
20 ~ reaction mixture was cooled to room temperature, then
reheated to reflux and 10 ml. of solvent distilled off.
Then 10 ml. of carbon tetrachloride was added dropwise near
the reflux temperature (77C.). The resulting mixture was
allowed to cool to room temperature and stirred Eor 1 hour.
The product was separated by filtration, washed with 14 ml.
of 40 percent carbon tetrachloride in 1,2-dichloroethane,
and dried at 50C. under vacuum, 2.12 grams of light colored
solid (86.5 percent yield). The NMR was identical with the
NMR on the product prepared as reported in Example 16.
X-4902A -77-
., , . ~ . ...
'., ' ' :'' ' " ' ' , . , . , ' ''
,
~3~L4~L
EXAMPLE 63: PREPARATION OF 3-(((3-(BENZYLOXYCARBONYLAMINO-
METHYL)-1,2,4-TRIAZOL-5-YL)THIO)METHYL)-7-(2-(2-THIENYL)-
ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid (2
grams; 5 mmole) and 3-(benzyloxycarbonylaminomethyl)-
1,2,4-triazole-5-thiol (2 grams; 7.6 mmole) in 35 ml. of
nitromethane were heated at 80-90C. for 6 hours, with
stirring. The reaction mixture was cooled and filtered to
separate the product. It was recrystallized twice from
aqueous acetone, 1.2 grams of cream colored crystals, m.p.
174-178C. (dec.) (40 percent yield). `
C25H24N6O6S3: C, 49.99; H, 4.03; N, 13.99;
S, 16.01.
Found : C, 50.20; H, 4.03; N, 13.76;
S, 15.68.
EXAMPLE 64: PREPARATION OF 3-(((l-(CARBOXYMETHYL)-lH-
TETRAZOL-5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN ACETONITRILE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(1.0 gram; 2.5 mmole) and l-(carboxymethyl)-lH-tetrazole-
`
5-thiol (0.61 gram; 3.8 mmole) in 75 ml. of acetonitrile was
heated to boiling and 35 ml. of the solvent distilled off.
The reaction mixture was then refluxed for 13 hours, cooled,
filtered, and concentrated under reduced pressure. The
residue was dissolved in ethyl acetate, washed with lN HCl
;~ and brine, and dried over sodium sulfate. Hexane was added,
resulting in a precipitate which was separated and triturated
with ether, 0.475 gram of tan solid (3~ percent yield). The
identity of the product was established by comparison with
an authentic sample prepared by another synthetic route.
X-4902A -78-
"
:~ .
. .~ . . ~ , ,
.. . . . . .
,, :: ~ ~. . .. .
~3~L4~L
EXAMPLE 65: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(3-CHLOROPROPIONAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN NITROMETHANE
A stirred suspension of 7-(3-chloropropionamido)-
cephalosporanic acid (1.0 gram; 2.5 mmole) and 5-methyl-
1,3,4-thiadiazole-2-thiol (0.4 gram; 3 mmole) in 20 ml. of
nitromethane was immersed in an oil bath at 95-96C.
According to TLC, reaction was complete in 3 hour~. The
reaction mixture was cooled to room temperature, filtered,
and evaporated under vacuum, leaving a light red oil which
crystallized on standing at room temperature for 2 hours.
It was then triturated with 15 ml. of ethyl acetate, filtered,
washed with ethyl acetate, and dried, 0.64 gram (55.2 percent
yield). Identity of the product was confirmed by IR, UV,
titration, microanalysis, and NMR.
C16HlgClN4O4S3: C, 41.51; H, 4.14; N, 12 10;
S, 20.78; Cl, 7.66.
Found : C, 41.70; H, 4.23; N, 11.84;
: :
S, 20.51; Cl, 7.88.
EXAMPLE 66: PREPARATION OF 3-(((1-BENZYL-lH-1,2,3-TRIAZOL-
5-YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-
4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(200 mg.; 0.5 mmole) and 1-benzyl-lH-1,2,3-triazole~5-
- thiol (140 mg.; 0.7 ~nole) were mixed in 15 ml. oE 1,2-
dichloroethane and heated at 65-70C. for 21 hours. Solvent
was removed under reduced pressure, 25 ml. of a saturated ;~
sodium bicarbonate solution was added, and the mixture was
extracted with two portions of ethyl acetate. Fresh ethyl
X-4902A -79-
: - ~ , : :
.:
: .
., .: ~ , . . :
3~4~
acetate was added to the remaining aqueous solution which
was cooled in an ice bath and the pH acljusted to 2.5 with 20
percent HC1 solution. The acidic prod~lct was extracted with
two portions of ethyl acetate, and the combined ethyl acetate
was washed with saturated NaCl solution and dried over
anhydrous magnesium sulfate. The magnesium sulfate was
filtered and ethyl acetate removed under reduced pressure~
76 mg. Thin layer chromatography showed it to be a mixture
of starting material and product by comparison with an
authentic sample of product and starting materials.
EXAMPLES 67-72: DISPLACEMENT BY VARIOUS THIOLS OF 7-
METHOXY-7-(2-(lH-TETR~ZOL-l-YL)ACETAMIDO)CEPHALOSPORANIC
ACID IN NITROMETHANE
7-Methoxy-7-(2-(lH-tetrazol-l-yl)acetamido)cephalo-
sporanic acid was reacted with various thiols in nitro-
methane. The procedures were essentially the same as
reported above, except that high pressure liquid chroma-
tography was employed to purify the products from unreacted
starting material. The products and identification were as
follows:
3-~((1-methyl-lH-tetrazol-5-yl)thio)methyl)-
7-methoxy-7-(2-(lH-tetrazol-l-yl)acetamido)-3-cep~em-4-
carboxylic acid, uv max (ethanol) 273 m~ (~ 9,082).
3-~((5 methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-
7-methoxy-7-(2~(1H-tetrazol-l-yl)acetamido)-3-cephem-4-
carboxylic acid, uv max (ethanol) 273 m~ ( E 12,785).
Anal-, Calc. for C15H16N8O5S3: C, 37.18; H, 3.33; N, 23.13.
Found : C, 36.99; H, 3.31; N, 22.86.
'~;
X-4902A -80-
.
~3~4~
3-(((5-methyl-1,3,4-oxadiazol-2-yl)thio)methyl)-
7-methoxy-7-(2-(lH-tetrazol-l-yl)acetamido)-3-cephem-~-
carboxylic acid, uv max (ethanol) 269 m~ (~ 8,910).
Anal., Calc. for C15H16N8O6S2
Found : C, 38.58; H, 3.69; N, 23.91.
3-(((1-(carboxymethyl)-lH-tetrazol-5-yl)thiol)-
methyl)-7-methoxy-7-(2-(lH-tetrazol-l-yl)acetamido)-3-
cephem-4-carboxylic acid, uv max (ethanol) 269 m~ (~ 7669).
Anal., Calc. for C15H16N10O7S2
Found : C, 35042; H, 3.38; N, 27.39.
3-(((4,5-dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-
triazin-3-yl)thio)methyl)-7-methoxy-7-(2-(lH-tetrazol-l-
yl)acetamido)-3-cephem-4-carboxylic acid, uv max (ethanol)
274 (~ 11,967).
Anal., Cala. for C16H17NgO7S2
Found : C, 36.96; H, 3.80; N, 27.63.
3-(((1,3,4-triazol-5-yl)thio)methyl)-7-methoxy-
7-(2-(lH-tetrazol-l-yl)acetamido)-3-cephem-4-carboxylic acid
Anal., Calc. for C14H15NgO5S2:
Found : C, 36.98; H, 3.43; N, 27.79.
EXAMPLE 73: PREPARATION OF 3-(((1-METHYL~lH-TETRAZOL-5-YL)-
.
THIO)METHYL)-7-(2-(4-ETHYL-2,3-DIOXO-l-PIPERAZINYLCARBONYL-
AMINO)-2-PHENYLACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN
NITROMETHANE -
7-(2-(4-Ethyl-2,3-dioxopiperazinylcarbonylamino)-
~ ~ 2-phenylacetamido)cephalosporanic acid hydrate (0.3 gram;
-~ 0.5 mmole) and 1-methyl~lH-tetrazole-5-thiol (0.0725 gram;
0.625 mmole) were added to 6 ml. of nitromethane which had
been dried over alumina and the mixture was heated at 85C.,
X-4902A -81- ; -
, . -, .
, ... . . . . . . . . .
.... , . . .. , , , :, , ,
33~
under nitrogen, for 12 hours. The nitromethane was then
evaporated and the residual brown foam redissolved in sodium
bicarbonate solution and washed twice with ethyl acetate.
Fresh ethyl acetate was added and the mixture cooled to 0C.
and adjusted to pH 2.3. The layers were separated and the
aqueous layer was washed with ethyl acetate. The ethyl
acetate portions were combined, washed with saturated NaCl
solution, dried over anhydrous magnesi~m sulfate, filtered,
and evaporated, yielding a pale yellow powder, 130 mg. ~40%
of theoretical). Identity of the product was confirmed by
NMR ~ 1.2 ~t, 3, -NCH2C_3, J = 7 Hz), 3.65 (m, 6, N-CH2CH3
and piperazinyl H), 4.0 (s, 3, -CH3 of tetrazole), 4.4 (s,
2, 3-C_2S-), 5.1 (d, 1, C6-_, J = 6 Hz), 5.8 (d, 1, C7-H,
o~
J - 8 Hz), 6.0 (d, 1, ~ / -CH- , J = 6 Hz), 7.4 (m, 5,
N
phenyl-_), 8.4 (d, 1, -CHCONH- , J = 8 Hz), and 10.0 (d,
1, -CHCONH- , J = 6 Hz).
N_
EXAMPLE 74: PREPARATION OF 3-(((l-CARBOXYMETHYL)-lH-TETRA-
ZOL-5-YL)THIO)METHYL)-7-(2-(4-ETHYL-2,3-DIOXO-l-PIPERAZINYL-
CARBONYLAMINO)-2-PHENYLACETAMIDO)-3-CEP~EM-4-CARBOXYLIC
ACID IN 1,2-DICHLOROETHANE ~
7-(2-(4-Ethyl-2,3-dioxopiperazinylcarbonylamino)- ~ `
2-phenylacetamido)cephalosporanic acid hydrate (0.3 gram; -
0.5 mmole) was dissolved in 45 ml. of 1,2-dichloroethane
which had been dried over alumina and the solution was
heated to 95 to azeotrope off water. After 40 ml. o~
solvent had been collected, nitromethane (10 ml.) and 1-
(carboxymethyl)-lH-tetrazole-5-thiol (0.160 gram; 1 mmole)
X-4902A -82-
- , . . . . ..
. ~ , . . . . .
3~
were added and the reaction mixture maintained at 90C. for
12 hours under nitrogen. The reaction mixture was then
filtered and solvent evaporated, yielding a gum to which
ethyl acetate and saturated sodium bicarbonate solution were
added. The mixture was washed twice with ethyl acetate,
then fresh ethyl acetate was added and the mixture cooled to
0C. and adjusted to pH 2.3 with 20% HCl. The layers were
separated and the aqueous layer was washed with ethyl acetate.
The ethyl acetate layers were combined and washed with
saturated HCl, dried over anhydrous magnesium sulfate,
filtered, evaporated, and dried at room temperature, 123 mg.
The identity of the product was confirmed by NMR, ~ 1.2 (t,
3, -NCH2C_3, J = 7 Hz), 3.6 (m, 4, N-CH~CH3 and piperazinyl
5-H), 4.1 (m, 2, piperazinyl 4-H), 4.4 (s, 2, 3-CH2S-~, 5.1
(d, 1, C6-H, J = 5 Hz), 5.3 (s, 2, tetrazole 1-CH2COOH),
5.75 (d, 1, C7-H, J = 6 Hz), 5.9 (d, 1, ~ CH- ), 7.4
_~ '
(m, 5, phenyl _), and 9.9 (d, 1, o~ ~--CH-, J = 7 Hz).
NH
EXAMPLE 75: PREPARATION OF 3-(((4,5-DIHYDRO-4-METH~L- ~ -
6-HYDROXY-5-OXO-1,2,4-TRIAZIN-3-YL)THIO)METHYL)-7-(2-(4-
ETHYL-2,3-DIOXO-1-PIPERAZINYLCARBONYLAMINO)-2-PHENYLACE-
TAMIDO)-3-CEPHEM-4-CAR~OX~LIC ACID IN NITROMETHANE
7-(2-(4-Ethyl-2,3-dioxo-1-piperazinylcarbonyl-
amino)-2-phenylacetamido)cephalosporanic acid hydrate (0.3
gram; 0.5 mmole) and 4,5-dihydro-4-methyl-6-hydroxy-
5-oxo-1,2,4-triazine-3-thiol (0.111 gram; 0.625 mmole) were
dissolved in 10 ml. of nitromethane under nitrogen at room
X-4902A -83-
:
. :,-. . :, . .
3~
temperature, and the reaction mixture heated at 85C. for
about 12 hours. A brown gummy precipitate formed and was
discarded. Solvent was evaporated slowly, and a yellow
precipitate formed. The residue was cooled, filtered, and
washed with diethyl ether, yielding two crops, identical on
TLC, of off-white solid. Total yield was 80 mg. (24 percent
yield). Identity of the product was confirmed by NMR ~ 1.1
(t, 3, -NCH2CH3, J = 6 Hz), 3.3 (s, 3, triazine-CH3), 3.65
(m, 4, N-CH2CH3 and piperazinyl 5-H), 4O0 (m, 2, piperazinyl
6-H), 4.6 (s, 2, 3-CH2S-), 5.1 (d, 1, C6-H, J = 5 Hz), 5.75 ;
(m, 2, ~ CH- and C7-_), 7.45 (m, 5, phenyl _), 9.5
(d, 1, -CHCONH-, J = 8 Hz) and 10.0 (d, 1, o~ CH- ,
-NH
J = Hz).
EXAMPLE 76: PREPARATION OF 3-(((5-METHYLTHIO-1,3,4-THIA- `
DIAZOL-2-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-l-YL)ACE~AMIDO)-
3-CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (15.0 grams; 39.2 mmole) and 5-(methylthio)-1,3,4-
thiadiazole-2-thiol (6.43 grams; 39.2 mmole) were suspended
in 500 ml. of nitromethane which had been passed through a
column of neutral alumina. The mixture was heated to 95C.
and the reaction mixture was maintained at 95C. for 6
hours, then cooled overnight, filtered, washed with 250 ml.
of nitromethane, and dried at 45C. for 4 hours, 18.08 gram
(94.9% yield).
X-4902A -84-
. , .
:.
- ~8~
This product was added to 150 ml. of water and
4.0 ml. of ac~tic acid. The pH was adjusted to 6.3 with lN
NaOH (required 107 ml.) and the mixture was stirred for 1
hour. A solution of 257 ml. of 60~ sodium lactate and
65 ml. of ethanol was then added, and the resulting mixture
allowed to stand for 45 minutes. It was then stirred for 45
minutes, filtered, washed three times with ethanol and
dried, 18.1 grams.
The product was reslurried in 300 ml. of ethanol
for 3 hours, filtered, washed with ethanol, and dried, 14.85
grams. The identity of the product was confirmed by HPLC. ~
EXAMPLES 77-84: PREPARATION OF 3-(((5-METHYL-1,3,4-THIA- -
DIAZOL-2-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-l-YL)ACETAMIDO)-
3-CEPHEM-4-CARBOXYLIC ACID IN VARIOUS SOLVENTS
7-t2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid was reacted with 5-methyl-1,3,4-thiadiazole-2-thiol in
various solvents. The key details of the reactions are ~ -
summarized below (all other aspects of the reaction being
eyplcal of the preceding examples).
'''" ~,
~ :.
'"
X-4902A -85-
'
, .. .' ' ~ . . , - .... , ., - .. ::
..
~ ^ .
~ u ra
n _ O
~,
u~
~ o u ~ ~ u
O O I a) O
0 o ~
` ll l l
:
c~o ~: :
~ h S l ~ ~h Ei
a~ o o o o o o o oo o
~ ,_11~ ~ 'I ' ' "
~,, '
.
~ ~C . , , . , , , , ~,
5~ ~ U ~ U O C~ ~ t)U
a) ~1 o o o o o o O o
~ ~ u~ O U~ O u~
E-
~
a) ~ ~ ,.
~ 0 ~
0 a) ~ ~ 0~ ~ o
4~ ~ ~ $ O ~ 0 ~
~) ^ O ~ ~ ~ .4~ O
F~ O ~ a)O ~ 0 ~a) h
~ ~ o ~ ~ 0~rl
r-l S J ~I N ~ 1 0 0a) O O ~ ~ -
o ~ a
u~ rl
~ us~ ~
o ~ o ~ o ~ ~ o
S~ I h Ql ~ ~~ h ~) a~
a) ,i ~ ~1 ~ ~ ~ o ~
rl h ~) ~ 0
rd ~ ~ ~I rl 0 1
o
.~ ~ . .
E~
r~
O ~ ~D 00
. ,s, a~ o ~` 0
o o o ,i o ,, o U~
;
o ;
: 0~ ~n :
~l rl
~C 0 ~ : . .
a) u
V ~ 0 ~
o s~ ~ ~ - ~ ~ Oa~
o
~ ,~ o ~ ,~
.
: ::
X-4 902A -86
. . :, .. , ., .. , : .
~ ' ' '~, . ' ,'. , ' ' ,. ' ' " ' ' ' '
EXAMPLE 85: PREPARATION OF LITHIUM 3-(((l-METHYL-lH-
TETRAZOL-5-YL)THIO)METHYL)-7-(2-HYDROXY-2-PHENYLACETAMIDO)-
3-CEPHEM-4-CARBOXYLATE IN GLACIAL ACETIC ACID
7-(2-Hydroxy-2-phenylacetamido)cephalosporanic
acid (0.48 gram; 1 mmole) and 1-methyl-lH-tetrazole-5-
thiol (0.5 gram; 4.3 mmole) were reacted in 15 ml. of glacial
acetic acid at 84-86C. for 8 hours. The reaction mixture
was cooled to room temperature and iodine t0.4 gram; 1.6
mmole) added to convert unreacted thiol to the disulfide.
The mixture was stirred 20 minutes at room temperature, then
poured into 100 ml. of ethyl acetate and 50 ml. oE water and
excess I2 removed by addition of sodium sulfite. The layers
were separated and the ethyl acetate la~er washed with two
100 ml. portions o~ water and one 50 ml. portion of 20% NaCl
solution and dried over anhydrous sodiwn sulfate. Ethyl
acetate was removed on a rotary evaporator, and the residue
dissolved in 10 ml. of methanol. Lithium acetate dihydrate
(0.21 gram; 2 mmole) was added and the mixture stirred for
20 minutes at room temperature. The product crystallized
and was separated by filtration, washed with 5 ml. of
methanol and dried, yielding 0.34 gram of white cyrstals
(69.4~ yield). The identity of the product was confirmed by
NMR: ~ 3.50 (ABq, J - 17 HZ), 3.92 (s, -C~l3 of tetrazole),
4.20 (s, 3-C_2S-), 5.04 (d, C6-_, J = 5 HZ), 5.24 (s,
0CHCONH-), 5.64 (d, C7-_, J = 5 Hz), and 7.44 (s, 0-H). - ~
The NMR spectrum was identical with an NMR spectrum on an
authentic sample of the product prepared by aqueous dis-
placement. -
X-4902A -87-
.~- ~.''' .
, '.~ . ' '. ,', ''.'.'. - ;. ' ' ' ' , ',
,. ' . ' . , ': ' ,
EXAMPLE 86: PREPARATION OF 3-(((l-METHYL-lH-TETRAZOL-
5-YL)THIO)METHYL)~7-(2-(o-(TERT-BUTOXYCARBONYLAMINOMETHYL)-
PHENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(o-(tert-Butoxycarbonylaminomethyl)phenyl)-
acetamido)cephalosporanic acid (0.2 gram; 0.386 mmole) and
1-methyl-lH-tetrazole-5-thiol (0.047 gram; 0.405 mmole) were
reacted in 5 ml. of nitromethane at 85C., under nitrogen,
for 12 hours, yielding 3-(((1-methyl-lH-tetrazol-5-yl)thio)-
methyl)-7-(2-to-(tert-butGxycarbonylaminomethyl)phenyl)-
acetamido)-3-cephem-4 cark,oxylic acid. Identity of the
product was confirmed by NMR, ~ 1.45 (s, 9, -C(CH3)3, 3.65
(s, 4, 2-CH2 and -CH2CONH-), 3.85 (s, 3, -C_3 of tetrazole),
4.3 (m, 4, 3-C_2S- and -CH2NHCOO-tert-C~Hg), 5.0 (d, 2,
C6-_, J = 4 Hz), 5.7 (q, 1, C7-H), 7.25 (s, 5, phenyl -H
and -CONH-), and 8.9 (s, 1, -COOH).
EXAMPLE 87: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-UREIDO-2-PHENYLACETAMIDO)-3-OEPHEM-
4-CARBOXYLIC ACID IN NITROMETHANE ~ `
7-(2-Ureido-2-phenylacetamido)cephalosporanic acid
20 (0.224 gram; 0.5 mmole) and 1-methyl-lH-tetrazole-5-thiol
(0.061 gram; 0.525 mmole) ~ere reacted in 8 ml. of nitro- ~ ;
methane at 85C~, for 12 hours, yielding 3-(((1-methyl-lH-
tetrazol-5-yl)thio)methyl)-7-(2-ureido-2-phenylacetamido)-
3-cephem-4-carboxylic acid. Identity of the product was
confixmed by NMR, ~ 3.5 (d, 2, 2-C_2, J = 3 Hz), 3.83 (s,
3, -CH3 of tetrazole), 4.17 (d, 2, 3-CH2S-, J = 3 Hz), 4.9
(d, 1, C6- , J = 5 Hz), 5.4 (d, 1, -CH2CONH-, J = B HZ) 5.6
(m, 3, C7-H and -NH2), 6.7 (d, 1, 0CH-, J = 8 Hz), 7.25 (m,
N_
5, phenyl H), and 9.2 (d, 1, -CH2CONH- , J = 8 I-lz).
X-4902A -88-
: .. , , , - . . :
... :, ,:, .. ... . . .
. .. .. . .
~383~1
EXAMPLE 88: PREPARATION OF 3-(((1-METElYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-~2-CYANOACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID IN NITROMETHANE
7-(2-Cyanoacetamido)cephalosporanic acid (0.339
gram; 1 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.122
gram; 1.05 mmole were reacted in 10 ml. of nitromethane at
85C., under nitrogen, for 12 hours, yielding 3-(((1-
methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-cyanoacetamido)-
3-cephem-4-carboxylic acid. Identity of the product was
confirmed by NMR, ~ 3.5 (s, 2, 2~CH2), 3.65 (s, 2, CNCH2CONH-),
3.95 (s, 3, -CH3 of tetrazole), 5.35 (s, 2, 3-C_2S-), 5.1
(d, 1, C6-_, J = 6 Hz), 5.75 (q, 1, C7-EI, J = 4 Hz, 7.8 (s,
1, -COO_), and 8.7 (d, 1, -CH2CON -, J = 9 Hz).
EXAMPLE 89: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(M-CHLOROPHENYLTHIO)ACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN NITROMETHANE
7-(2-(m-Chlorophenylthio)acetamido)cephalosporanic
acid (0.390 gram; 0.855 mmole) and l-methyl-lH-tetrazole-
5-thiol (0.104 gram; 0.9 mmole) were reacted in 10 ml. of
nitromethane at 85C., under nitrogen, for 12 hours, yielding
3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-(m-chloro- -
phenylthio)acetamido)~3-cephem-4-carboxylic acid. Identity
of the product was con~irmed by NMR ~ 3.6 (s, 2, Z-CH2, 3.75
(s, 2, -C_2CONH-), 3.9 (s, 3, -CH3 of tetrazole), 4.4 (s, 2, ~ ~
3-CH2S-), 5.0 (d, 1, C6-H, J = 5 Hz), 5.8 (q, 1, C7-_, J = ~ ~ -
4 Hz), 7.15 (m, 4, phenyl-H), 7.7 (d, 1, -CH2CONH-, J =
8 Hz), and 8.5 (s, 1, COO ).
,, '.
.
X-4902A -89-
3~
EXAMPLE 90: PREPARATION OF 3-(((l-MErrHYL-lH-TETRAZOL 5-
YL)THIO)METHYL)-7-(2-(PHENYLTHIO)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN NITROMETHAME
7-(2-Phenylthio)acetamido)cephalosporanic acid
(0.422 gram; 1 mmole) and 1-methyl-lH~tetrazole-5~thiol
(0.122 gram; 1.05 mmole) were reacted in 10 ml. of nitro-
methane at 85C., under nitrogen, for 12 hours, yielding
3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-(phenyl-
thio)acetamido)-3-cephem-4-carboxylic acid. Identity of the
product was confirmed by NMR, ~ 3.7 (s, 4, -CH2CONH- and
2-CH2), 3.9 (s, 3, -CH3 of tetrazole), 4.35 ~s, 2, 3-CH2S-),
4.9 (d, 1, C6-H, J = 6 Hz), 5.8 (q, 1, C7-H, J = 4 Hz), 7.25
(s, 5, phenyl-H), 7.7 (d, 1, -CH2CONH-, J = 8 Hz), and 9.4 ~-
(s, 1, -COOH). ;
EXAMPLE 91: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN n-BlJTYLFORMATE
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
(3.96 gram; 10 mmole) and 1-methyl-lH-tetrazole-5-thiol
(1.28 gram, ll mmole) were reacted in 98 ml. of n-butyl
formate at 84-86C. for 48 hours, yielding 3-(((1-methyl-
lH-tetrazol-5-yl)thio)methyl)-7-(2-(2-thienyl)acetamido)-
3-cephem-4-carboxylic acid, m.p. 167.5-168C., yield 79.9%.
The NMR spectrum on the product was identical with the NMR
spectrum on the product of Example 5. -
.
' .
X-4902A -90-
'
.' ~ '''' ' : " ' '
~: ,' ,~ , 1 ,
4~
EXAMPLE 92: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(2-(2-THIENYL)ACETAMIDO)-3-CEPHEM-4-
CARBOXYLIC ACID IN TOLUENE
The reaction reported in Example 91 was repeated
on the same scale but in 150 ml. of toluene at 84-86C., for
136 hours. The product melted at 163-163.5C. and was
obtained in 88.5% yield. The NMR spectrum on the product
was identical with the NMR spectrum on the product of
Example 5.
EXAMPLE 93: PREPARATION OF 3-(((1-METHYL-lH-TETRAZOL-5-
YL)THIO)METHYL)-7-(tert-BUTOXYCARBONYLAMINO)-3-CEPHEM-4-
CARBOXYLIC ACID IN NITROMETHANE
7-(tert-Butoxycarbonylamino)cephalosporanic acid
(0.372 gram; 1 mmole) and 1-methyl-lH-tetrazole-5-thiol ~-
(0.122 gram; 1.05 mmole) were reacted in 10 ml. of nitro- ,
methane at 80C. for 12 hours, yielding 3-(((1-methyl- ~
~, ,
lH-tetrazol-5-yl)thio)methyl)-7-(tert-butoxycarbonylamino) ; '
3-cephem-4-carboxylic acid. The identity of the produ~ ~was
confirmed by NMR, ~ 1.45 (s, 9, -C(C_3)3), 3.7 (9
2-CH2), 3.95 (s, 3, -CH3 of tetrazole), 4.4 (s ~2, 3-CH2S-),
5.0 (s, 1, C6-H), 5.6 (s, 2, C7-H and ~CH2CON_), and 13 (s,
1, -COOH).
EXAMPLE 94: PREPARATION OF 3-(((1-METHYL-lH-TET~AZOL-5-
YL)THIO)METHYL)-7-(2-(4-ETHYL-2,3-DIOXOPIPERAZINYLCARBONYL-
AMINO)-2-(p-HYDROXYPHENYL)ACETAMIDO)-3-CEPHEM-4-CARBOXYLIC
ACID
7-(2-(4-Ethyl-2,3-dioxopiperazinylcarbonylamino)-
2-(p-hydroxyphenyl)acetamido)cephalosporanic acid (0.180
gram; 0.306 mmole) and 1-methyl-lH-tetrazole-5-thiol (0.0374
-
X-4902A -91-
~ : - . .. .
. i , :, :: .
, ., , , , , " , . . . . . .
, .;, . . . . . ... .
gram; 0.322 mmole) were reacted in 6 ml. of nitromethane at
80C. for 12 hours, yielding 3-(((1-methyl-lH-tetrazol-
5-yl)thio)methyl)-7-(2-(4-ethyl-2,3-dioxopiperazinylcar-
bonylamino)-2-(p-hydroxyphenyl)acetamido)-3-cephem-4-
carboxylic acid. The identity of the product was confirmed
by N~R, ~ 1.2 (t, 3, N-CH2CEI3), 3.6 (m, 6, 2-CH2, piperazinyl
6-_ and N-C_2CH3), 3.9 (m, 2, piperazinyl 5-_), 3.95 (s,
3, -CH3 of tetrazole), 4.4 (s, 2, 3-C_2S-), 5.0 (d, 1,
C6-H, J = 6 Hz), 5.6 (d, 1, C7-_, J = 6 Hz), 5.8 (d, 1,
10 CHCONH-, J = 8 Hz), 6.8 (d, 2, phenyl-H, J = 8 Hz), 7.3 (d,
2, phenyl-H, J = 8 Hz), 8.3 (d, 1, -CHCON_-, J = 8 Hz), and
9.9 (d, 1, -CHCONH-, J = 6 Hz.
-N_
EXAMPLE 95: PREPARATION OF 3-(((1,3,4-THIADIAZOL-2-YL)THIO)
METHYLj-7-(2-(lH-TETRAZOL-l-YL)ACETAMIDO)-3-CEPHEM-4-CAR-
BOXYLIC ACID IN ACETONITRILE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
; acid (0.38 gram; 1 mmole) and 1,3,4-thiadiazole-2-thiol
(0.15 gram; 1.27 mmole) were reacted in 15 ml. of aceto-
nitrile at reflux, under nitrogen, for 24 hours, yielding
3-(((1,3,4-thiadiazol-2-yl)thio)methyl)-7-(2-(lH-tetrazol-
l-yl)acetamido)-3-cephem-4-carboxylic acid. The identity of
the product was confirmed by NMR, ~ 3.74 (m, 2, 2 CH2-),
; 4-48 (q, 2, 3-CH2S-, JAB = 14), 5-16 (d, 1, C6-H, J = 5),
5.42 (s, 2, -CH3 of tetrazolyl), 5.76 (q, 1, C7-H, J6 7 ~
5' J7NH = 8), 8.87 (s, 1, tetrazole ring), 9.40 (s, 1, thia-
diazolethiol starting material), 9.53 (d, 1, -CH2CONH-, J =
8 Hz), and 9.57 (s, 1, thiadiazole riny).
,
X-4902A -92-
.. . . .
~ 33~
EXAMPLE 96: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(5-CARBOMETHOXY-5-(2,4-DICHLOROBENZ-
AMIDO)VALERAMIDO)-3-CEPHEM-4-CARBOXYLIC ACID IN 1,2-
DICHLOROETHANE
7-(5-Carbomethoxy-5-(2~4-dichlorobenzamido)valer-
amido)cephalosporanic acid (0.3 gram; 0.5 mmole) and 5-
methyl-1,3,4-thiadiazole-2-thiol (0.1 gram; 0.75 mmole) were
reacted in l,2-dichloroethane at reflux for 16 1/2 hours,
yielding 3-(((5-methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-
7-(5-carbomethoxy-5-(2,4-dichlorobenzamido)valeramido)-
3-cephem-4-carboxylic acid.
EXAMPLE 97: PREPARATION OF LITHIUM 3-(((1-METHYL-lH-
TETRAZOL-5-YL)THIO)METHYL)-7-(2-(lH-TETRAZOL-l-YL)ACET-
AMIDO)-3-CEPHEM-4-CARBOXYLATE IN ACETONITRILE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (1.91 grams; 5 mmole) and 1-methyl-lH-tetrazole-5-
thiol (0.7 gram; 6 mmole) in 50 ml. of acetonitrile were
refluxed for 24 hours and cooled to room temperature.
Solvent was removed on a rotary evaporator, to about 10 ml.
Ethanol (40 ml.) was added; followed b~ a solution of 0.3
gram of lithium hydroxide in 10 ml. of methanol. The
product, lithium 3-1((1-methyl-lH-tetrazol-5-yl)thio)methyl)-
7-(2-(lH-tetrazol-l-yl)acetamido)-3-cephem-4-carboxylate,
crystallized. After the reaction mixture was stirred for 45
mi~utes at room temperature, the product was separated by
filtration, washed with ethanol, and vacuum dried at 40C.,
1.60 grams (72% yield). rrhe identity ~f the product was
confirmed by NMR.
X-4~02~ -93-
~3~4~L
EXAMPLE 98: PREPARATION OF 3-(((5-METHYL-1,3,4-THIADIAZOL-
2-YL)THIO)METHYL)-7-(2-FORMYLOXY-2-PHENYLACETAMIDO)-3-
CEPHEM-4-CARBOXYLIC ACID IN 1,2-DICHLOROETHANE
7-(2-Formyloxy-2-phenylacetamido)cephalosporanic
acid (4.6 grams; 8.8 mmole) and 5-methyl-1,3,4-thiadiazole-
2-thiol (1.52 grams; 11.5 mmole) in 50 ml. of 1,2-dichloro-
ethane were refluxed for 12 hours and cooled to room tem-
perature. The product, 3-(((5-methyl-1,3,4-thiadiazol-
2-yl)thio)methyl)-7-(2-formyloxy-2-phenylacetamido)-3-
cephem-4-carboxylic acid, precir)itated as a thick paste.
The mixture was diluted with an additional 50 ml. of 1,2-
dichloroethane and stirred 4 hours at room temperature. The
product was then separated by filtration, washed with 1,2- -~
dichloroethane until the filtrate was clear, and vacuum
dried at 40-45C., 2.8 grams (63% yield). The identity of
the product was confirmed by NMR.
EXAMPLE 99: PREPARATION OF 3-(((4,5-DIH~DRO-6-HYDROXY-4-
M~THYL-5-OXO-1,2,4-TRIAZIN-3-YL)THIO)METI~YL)-7-(2-(lH-
TETRAZOL-l-YL)ACETAMIDO)-3-CEPH~M-4-CARBOXYLIC ~CID IN
ACETONITRILE
7-(2-(lH-Tetrazol-l-yl)acetamido)cephalosporanic
acid (8.9 grams; 23.2 mmole) and 4,5-dihydro-6-hydroxy-4-
methyl-5-oxo-1,2,4-triazine-3-thiol (4.1 grams; 25.8 mmole)
in 200 ml. of acetonitrile were refluxed 23 hours then
cooled to room temperature. Product had crys~allized during
the course of the reaction and was separated by filtration,
washed with 50 ml. of acetonitrile, and vacuum dried at
40-50C., 9.70 grams (86.6~ yield). The identity of the
produc-t 3-(((4,5-dihydro-6-hydroxy-4-methyl-5-oY~o-1,2,4-
30 triazin-3-yl)thio)methyl)-7-(2-(1ll-tetrazol-1-yl)acetamido)-
3-cephem-4-carboxylic acid, was confirmed by NMR.
X-4902A -94-
:' '
.. . . . .
3~
EXAMPLES 100-144
Yet other reactions in accordance with the present
process can be conducted as ~ollows:
7-(2-Sulfo-2-phenylacetamido)cephalosporanic acid~
sodium salt, is reacted with l-methyl-lH-tetrazole-5-
thiol, yielding 3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-
7-(2-sulfo-2-phenylacetamido)-3-cephem-4-carboxylic acid,
sodium salt.
7-(2-(4-Pyridylthio)acetamido)cephalosporanic acid
is reacted with 1-methyl-lH-tetrazole-5-thiol, yielding
3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-(4- ~ -
pyridylthio)acetamido)-3-cephem-4-carboxylic acid.
7-(5-Carbo-n-butoxy-5-(2,4-dichlorobenzamido)-
valeramido)cephalosporanic acid and 5-methyl-1,3,4-thia-
diazoIe-2-thiol are reacted, yielding 3-(((5-methyl-1,3,4-
thiadiazol-2-yl)thlo)methyl)-7-(5-carho-n-butoxy-5 (2,4-
dichlorobenzamido)valeramido)-3-cephem-4-carboxylic acid.
7-(5-Carbo-n-butoxy-5-(2,4-dichlorobenzamido)-
~, ...... ..
valeramido)cephalosporanic acid and l-methyl-lH-tetrazole-
5-thiol are reacted yielding 3-(((1-methyl-lH-tetrazol-
5-yl?thio)methyl)-7-(5-carbo-n-butoxy-5-(2,4-dichlorobenz-
amido)valeramido)-3-cephem-4-carboxyli.c acid.
7-(2-(2-(p-Nitrobenzyloxycarbonylamino)thiazol-
~, .
4-yl)-2-(methoxyimino)acetamido)cephalosporanic acid is
reacted with l-methyl-lH-tetrazole-5-thiol, yielding 3-
(((l-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-(2-(p-
nitrobenzyloxycarbonylamino)thiazol-4-yl)-2-(methoxyimino)-
acetamido)-3-cephem-4-carboxylic acid.
, ~ . ...
.:. . . ..
X-4902A -95-
"~', .'.
'' - , .
', . ~ , , , , . . , . , , ~
: -, ' ' ' ' . '; . . . , '': ' . , ' ' ' , , . .. ': . .
,, . ~. : . ~ .. : . . . . . .
.. : .. .. .. . . . . .. ..
~ 33~
7-(2-(2-(p-Nitrobenzyloxycarbonylamino)thiazol-
4-yl)acetamido)cephalosporanic acid is reacted with l-methyl-
lH tetrazole-5-thiol yielding 3-(((1-methyl-lH-tetrazol-5-
yl)thio)methyl)-7-(2-(2-(p-nitrobenzyloxycarbonylamino)-
thiazol-4-yl)acetamido)-3-cephem-4-carboxylic acid.
7-((2,2,2-Trichloroethoxy)carbonylamino)cephalos-
poranic acid is reacted with 6-hydroxypyridazine-3-thiol,
yielding 3-(((6-hydroxypyridazin-3-yl)thio)methyl)-7-
((2,2,2-trichloroethoxy)carbonylamino)-3-cephem-4-carboxylic
acid.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid is
reacted with 6-hydroxypyridazine-3-thiol, yielding 3-
(((6-hydroxypyridazin-3-yl)thio)methyl)-7-(2-(2-thienyl)-
acetamido)-3-cephem-4-carboxylic acid.
7-((2,2,2-Trichloroethoxy)carbonylamino)cephalos-
poranic acid is reacted wlth tetrazolo[l,5-b]pyridazine-6-
thiol, yielding 3-(((tetrazolo[1,5-b]pyridazin-6-yl)thio~
methyl)-7-((2,2,2-trichloroethoxy)carbonylamino)-3-cephem-
4-carboxylic acid. ~-
7-(2-(2-Thienyl)acetamido)cephalosporanic acid is
reacted with tetrazolo[l,5-b]pyridazine-6-thiol, yielding
3-(((tetrazolo[1,5-b]pyridazin-6-yl)thio)methyl)-7-(2-
(2-thienyl)acetamido)-3-cephem-4-carboxylic acid.
7-((2,2,2-Trichloroethoxy)carbonylamino)cephalos-
poranic acid is reacted with l-(sulfomethyl)-l~l-tetrazole-
5-thiol, sodium salt, yielding 3-(((1-(sulfomethyl)-1~1- ; -
tetrazol-5-yl)thio)methyl)-7-((2,2,2-txichloroethoxy)car-
bonylamino)-3-cephem-4-carboxylic acid, sodium salt.
~'. ".
X-4902A -96-
:
.:
-- ~L0~33~
7-(2-(2-Thienyl)acetamido)cephalosporanic acid is
reacted with l-(sulfomethyl)-lH-tetrazole-5-thiol, sodium
salt, yielding 3-(((1-(sulfomethyl)-lH-tetrazol-5-yl)thio~-
methyl)-7-(2-(2-thienyl)acetamido)-3-cephem-4-carboxylic
acid, sodium salt.
7-((2,2,2-Trichloroethoxy)carbonylamino)cephalos-
poranic acid is reacted with l-(2-(dimethylamino)ethyl)-
lH-tetrazole-5-thiol, yielding 3-(~ (2-(dimethylamino)-
ethyl)-lH-tetrazol-5-yl)thio)methyl)-7-((2,2 ! 2-trichloro-
ethoxy)carbonylamino)-3-cephem-4-carboY~ylic acid.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid -
is reacted with l-(2-(dimethylamino)ethyl)-lH-tetrazole-5- -
thiol, yielding 3-(((1-(2-(dimethylamino)ethyl)-lH-tetrazol-
5-yl)thio)methyl)-7-(2-(2-thienyl)acetamido)-3-cephem-4-
carboxylic acid.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
is reacted with p-methoxybenzenethiol, yielding 3-(((p-
methoxyphenyl)thio)methyl)-7-(2-(2-thienyl)acetamido)-
3-cephem-4-carboxylic acid.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid
is reacted with p-chlorobenzenethiol, yielding 3-(((p-chloro-
phenyl)thio)methyl)-7-(2-(2-thienyl)acetamido)-3-cephem-4-
carboxylic acid.
7-(2-(2-Thienyl)acetamido)cephalosporanic acid is
reacted with o-toluenethiol, yielding 3-(((o-tolyl)thio)-
methyl)-7-(2-(2-thienyl)acetamido)-3-cephem-4-carboxylic
acid.
.: '
X-4902~ _97_
.. . .
. .
~ .
7-(2-(lH-Tetrazol-l-yl~acetamido)cephalosporanic
acid is reacted with 1,3,4-thiadiazole-2-thiol, yielding
3-(((1,3,4-thiadiazol-2-yl)thio)methyl)-7-(2-(1l~-tetrazol-
l-yl)acetamido)-3-cephem-4-carboxylic acid.
7-Phthalimidocephalosporanic acid is reacted with
5-methyl-1,3,4-thiadiazole-2-thiol, yielding 3-(((5-methyl-
1,3,4-thiadiazol-2-yl)thio)methyl)-7-phthalimido-3-cephem-
4-carboxylic acid.
7-~2-Hydroxy-2-phenylacetamido)cephalosporanic
acid is reacted with 5-methyl-1,3,4-thiadiazole-2-thiol,
yieIding 3-(((5-methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-
7-(2-hydroxy-2-phenylacetamido)-3-cephem-4-carboxylic acid.
7-(2-Acetoxy-2-phenylacetamido)cephalosporanic
acid is reacted with 5-methyl-1,3,4-thiadiazole-2-thiol,
yielding 3-(((5-methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-
7-(2-acetoxy-2-phenylacetamido)-3-cephem-4-carboxylic acid.
7-(2-(Trifluoromethylthio)acetamido)cephalosporanic
acld is reacted with l-methyl-lH-tetrazole-5-thiol, yielding
3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-(trifluoro-
methylthio)acetamido)-3-cephem-4-carboxylic acid.
7-(2-(2-(p-Nitrobenzyloxycarbonylamino)thiazol-
4-yl)-2-(methoxyimino)acetamido)cephalosporanic acid
is reacted with 4,5-dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-
triazine-3-thiol, yielding 3-(((4,5-dihydro-6-hydroxy-4-
methyl-5-oxo-1,2,4-triazin-3-yl)thio)methyl)-7-(2-(2-(p-
nitrobenzyloxycarbonylamino)-3-cephem~4-carboxylic acid.
7-(2-(2-(p-Nitrobenzyloxycarbonylamino)thiazol-4-
yl)acetamido)cephalosporanic acid is reacted with 1-(2-
(dimethylamino)ethyl)-lH-tetrazole-S-thiol, yielding 3-
X-4902A -9~-
' '
. : . .. . . , -., ,: . '
' ~
~t33~
~ (2-(dimethylamino)ethyl)-lH-tetrazol-5-yl)thio)methyl)-
7-(2-(2-(p-nitrobenzyloxycarbonylamlno)thiazol-4-yl)acet-
amido)-3-cephem-4-carboxylic acid.
7-(2-(o-(tert-Butoxyearbonylaminomethyl)phenyl)-
acetamido)cephalosporanic acid is reacted with tetrazolo-
[1,5-b]pyridazine-6-thiol, yielding 3-(((tetrazolo[1,5-b]- -
pyridazin-6-yl)thio)methyl)-7-(2-(o-(tert-butoxycarbonyl-
aminomethyl)phenyl)aeetamido)-3-eephem-4-carboxylic acid.
7-(2-Formyloxy-2-phenylaeetamido)eephalosporanic
acid is reaeted with 1-(sulfomethyl)-lH-tetrazole-5-thiol,
sodium salt, yielding 3-(((1-(sulfomethyl)-lH-tetrazol-5-
yl)thio)methyl)-7-(2-formyloxy-2-phenylacetamido)-3-
cephem-4-earboxylie aeid.
7-(2-Hydroxy-2-phenylaeetamido)eephalosporanie
aeid is reaeted with l-(sulfomethyl)-lH-tetrazole-5-thiol,
sodium salt, to produee 3-(((1-(sulfomethyl)-111-tetrazol-
5-yl)thio)methyl)-7-(2-hydroxy-2-phenylacetamido)-3-
eephem-4-earboxylie aeid.
7-(2-(2-Thienyl)aeetamido)eephalosporanic aeid is
reaeted with 2-oxazolethiol to yield 3-(((oxazol-2-yl)-
thio)methyl)-7-(2-(2-thienyl)acetamido)-3-cephem-4-car-
boxylic acid.
7-(2-Formyloxy-2-phenylaeetamido)eephalosporanie
aeid and l-(earboxymethyl)-lH-tetrazole-5-thiol are reaeted
to produee 3-(((1-(earboxymethyl)-lH-tetrazol-5-yl)thio)-
methyl)-7-(2-formyloxy-2-phenylaeetamido)-3-eephem-4-
earboxylie aeid.
~ ' :
~: ' .,
X-4902~ -99-
7-(2-(Trifluoromethylthio)acetamido)cephalosporanic
acid and l-methyl-lH-tetrazole-5-thiol are reacted to
prepare 3-(~(l-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2~
(trifluoromethylthio)acetamido)-3-cephem-4-carboxylic acid.
7-(2-(Cyanomethylthio)acetamido)cephalosporanic
acid is reacted with l-methyl-lH-tetrazole-5-thiol to
produce 3-(((l-methyl-lH-tetrazol-5-yl)thio)methyl)-7-
(2-(cyanomethylthio)acetamido)-3-cephem-4-carboxylic acid.
7-Methoxy-7-(2-(cyanomethylthio)acetamido)cepha-
losporanic acid is reacted with l-methyl-lH-tetrazole-
5-thiol to obtain 3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-
7-methoxy-7-(2-(cyanomethylthio)acetamido)-3-cephem~4-
carboxylic acid.
7-(2-(o-Benzoylaminomethylphenyl)acetamido)cepha-
losporanic acid and l-(carboxymethyl)-lH-tetrazole-5-thiol ~;
are reacted to prepare 3-(((1-(carboY~ymethyl)-lH-tetrazol-
5-yl)thio)methyl)-7-(2-(o-benzoylaminomethylphenyl)acet-
amido)-3-cephem-4-carboxylic acid.
7-(2-Ureido-2-thienylacetamido)cephalosporanic
acid is reacted with 1-methyl-lH-tetrazole-5-thiol to obtain
3-(((1-methyl-lH-tetrazol-5-yl)thio)methyl)-7-(2-ureido-
2-thienylacetamido)-3-cephem-4-carboxylic acid.
7-(2-(2-Amino-4-thiazolyl)acetamido)cephalosporanic
; acid is reacted 1-(2-(dimethylamino)ethyl)-lH-tetrazole-
5-thiol to produce 3-(((1-(2-dimethylamino)ethyl)-lH-
tetrazol-5-yl)khio)methyl)-7-(2 (2-amino-4-thiazolyl)acet-
amido)cephalosporanic acid.
X-4902A -100-
.. . . .
. . . :
~as3~41
7 (5-senzoylamino-5-carboxyvaleram:i~o)ceplla-
losporanic acid is reacted with 4,5-dihydro-6-hydroxy-4-
methyl-5-oxo-1,2,4-triazine-3-thiol to prepare 3-(((4,5-
dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-triazin-3-yl)thio)-
methyl)-7-(5-benzoylamino-5-carboxyvaleramido)-3-cephem-
4-carboxylic acid.
7-(5-Benzoylamino-5-carboxyvaleramido)cephalosporanic
acid is reacted with l-methyl-lH-1,2,3-triazole-5-thiol to
prepare 3-(((1-methyl-lH-1,2,3-triazol-5-yl)thio)methyl)-
7-(5-benzoylamino-5-carboxyvaleramido)-3-cephem-4-carboxylic
acid.
7-(5-Benzoylamino-5-carboxyvaleramido)cephalosporanic
acid and l-(carboxymethyl)-lH-tetrazole-5-thiol are reacted - -
to produce 3-(((1-(carboxymethyl)-lH-tetrazol-5-yl)thio)-
methyl)-7-(5-benzoylamino-5-carboxyvaleramido)-3-cephem-
4-carboxylic acid.
7-(5-Benzoylamino-5-carboxyvaleramido)cephalosporanic
acid is reacted with 5-methyl-1,3,4-oxadiazole-2-thiol to ~
obtain 3-(((5-methyl-1,3,4-oxadiazol-2-yl)thio)methyl)-7- -
(5-benzoylamino-5-carboxyvaleramido)-3-cephem-4-carboxylic
acid.
7-(5-Benzoylamino-5-carboxyvaleramido)cephalosporanic
acid is reacted with l-b~nzyl-lH-tetrazole-5-thiol to obtain
3-(((1-benzoyl-lH-tetrazol-5-yl)thio)m~thyl)-7-(5-benzoyl-
amino-5-carboxyvaleramido)cephalosporallic acid.
7-(5-Benzoylamino-5-carboxyvaleramido)cepha- -~
losporanic acid is reacted with thiourea to obtain 3-
amidinothiomethyl-7-(5-benzoylamino-5-carboxyvaleramido)-
3-cephem-4-carboxylic acid.
X-4902A -101-
. : . . . -: : . :
:' ' ' ~ ' ~ ' ' '' : :
- . . : ,
:- . :
~3~
7-Methoxy-7-(5-benzoylamino-5-carboxyvaleramido)-
cephalosporanic acid is reacted with l-methyl-lH-tetrazole-
5-thiol -to obtain 3-(((1-methyl-lH-tetrazol-5-yl)thio)-
methyl)-7-methoxy-7-(5-benzoylamino-5-carboxyvaleramido)-
3-cephem-4-carboxylic acid.
7-Methoxy-7-(5-benzoylamino-S-carboxyvaleramido)-
cephalosporanic acid is reacted with 5-methyl-1,3,4-
thiadiazole-2-thiol to obtain 3-(((5-methyl-1,3,4-thia-
diazol-2-yl)thio)methyl)-7-methoxy-7-(5-benzoylamino-5-
I0 carboxyvaleramido)-3-cephem-4-carboxylic acid.
7-Methoxy-7-(5-benzoylamino-5-carboxyvaleramido)-
cephalosporanic acid is reacted with 4,5-dihydro-6-hydroxy- -
4-methyl-5-oxo-1,2,4-triazine-3-thiol to obtain 3-(((4,5-
dihydro-6-hydroxy-4-methyl-5-oxo-1,2,4-triazin-3-yl)thio)-
methyl)-7-methoxy-7-(5-benzoylamino-5-carboxyvaleramido)-
3-cephem-4-carboxylic acid.
7-Methoxy-7-(5-benzoylamino-5-carboxyvaleramido)-
cephalosporanic acid and thiourea are reacted to obtain 3-
amidinothiomethyl-7-methoxy-7-(5-benzoylamino-5-carboxy-
valeramido)-3-cephem-4-carboxylic acid.
7-(5-Butoxycarbonylamino-5-carboxyvaleramido)-
3-carbamoyloxymethyl-3-cephem-4-carboxylic acid is reacted
with thiourea to prepare 3-amidinothiomethyl-7-(5-butoxy-
carbonylamino-5-carboxyvaleramido)-3-cephem-4-carboxylic
acid.
7-(5-Butoxycarbonylamino-5-carboxyvaleramido)-
3-carbamoyloxymethyl-3-cephem-4-carboxylic acid is reacted
with 1-methyl-lH-tetrazole-5~thiol to obtain 3-(((1-
rnethyl-ll-l-tetrazol-5-yl)thio)methyl)-7-(5-bu~o~carborlyl-
arnino-5-carboxyvaleramido)-3-cephem-4-c.lr~)oxyl:ic ~c:id.
X-4902A -102-
,' , ~. ,.
.
:, . . . . . .... .... . . . .
': '.. ,: " . ':.. ' "' ' ' . ~ .
., . , . :, -
4~
7-(5-Butoxycarbonylamino-5-carboxyvaleramido)-3-
carbamoyloxymethyl-3-cephem-4-carboxylic acid and 5-methyl-
1,3,4-thiadiazole-2-thiol are reacted to prepare 3-(((5-
methyl-1,3,4-thiadiazol-2-yl)thio)methyl)-7-(5-butoxy-
carbonylamino-5-carboxyvaleramido)-3-cephem-4-carboxylic
acid.
7-(5-Butoxycarbonylamino-5-carboxyvaleramido)-
3-carbamoyloxymethyl-3-cephem-4-carboxylic acid and 1-
(carboxymethyl)-lH-tetrazole-5-thiol are reacted to prepare
10 3-(((1-(carboxymethyl)-lH-tetrazol-5-yl)thio)methyl)-7- ~.
(5-butoxycarbonylamino-5-carboxyvaleramido)-3-cephem-4-
carboxylic acid. ~::
~, .
'
: ~ .
~;
:,'' '
',,
~ X-4902A -103-
,
' ', , ,
