CA1062701A - Cephalosporins - Google Patents
CephalosporinsInfo
- Publication number
- CA1062701A CA1062701A CA231,504A CA231504A CA1062701A CA 1062701 A CA1062701 A CA 1062701A CA 231504 A CA231504 A CA 231504A CA 1062701 A CA1062701 A CA 1062701A
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- Prior art keywords
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- acid
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D277/00—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings
- C07D277/02—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings
- C07D277/20—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
- C07D277/32—Heterocyclic compounds containing 1,3-thiazole or hydrogenated 1,3-thiazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D277/56—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D271/00—Heterocyclic compounds containing five-membered rings having two nitrogen atoms and one oxygen atom as the only ring hetero atoms
- C07D271/02—Heterocyclic compounds containing five-membered rings having two nitrogen atoms and one oxygen atom as the only ring hetero atoms not condensed with other rings
- C07D271/08—1,2,5-Oxadiazoles; Hydrogenated 1,2,5-oxadiazoles
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Cephalosporin Compounds (AREA)
Abstract
ANTIBACTERIAL AGENTS
Abstract of the Disclosure 7-[D-(.alpha.-Amino-a-phenyl-, 2-thienyl-and 3-thienylacetamido]-3-(thiazol-2-yl)carbonyl-thiomethyl-3-cephem-4-carboxylic acid and 7-[D-(.alpha.-amino-a-phenyl-, 2-thienyl- and 3-thienylacetamido]-3-(5-methylthiazol-2-yl)carbon-ylthiomethyl-3-cephem-4-carboxylic acid and 7-[D-(.alpha.-Amino-.alpha.-phenyl-, 2-thienyl- and 3-thienylacetamido]-3-(3-methyl-1,2,5-oxadiazol-4-yl)-carbonylthiomethyl-3-cephem-4-carboxylic acids and their pharmaceutically acceptable salts are valuable as antibacterial agents, as nutritional supplements in animal feeds, as agents for the treatment of mastitis in cattle and as therepeutic agents in poultry and animals, including man.
The compounds are especially useful in the treat-ment, particularly by oral administration, of infectious diseases caused by many Gram-positive and Gram-negative bacteria. Also included within the invention are easily cleavable esters of the above acids and pharmaceutically acceptable acid addition salts of said esters.
Abstract of the Disclosure 7-[D-(.alpha.-Amino-a-phenyl-, 2-thienyl-and 3-thienylacetamido]-3-(thiazol-2-yl)carbonyl-thiomethyl-3-cephem-4-carboxylic acid and 7-[D-(.alpha.-amino-a-phenyl-, 2-thienyl- and 3-thienylacetamido]-3-(5-methylthiazol-2-yl)carbon-ylthiomethyl-3-cephem-4-carboxylic acid and 7-[D-(.alpha.-Amino-.alpha.-phenyl-, 2-thienyl- and 3-thienylacetamido]-3-(3-methyl-1,2,5-oxadiazol-4-yl)-carbonylthiomethyl-3-cephem-4-carboxylic acids and their pharmaceutically acceptable salts are valuable as antibacterial agents, as nutritional supplements in animal feeds, as agents for the treatment of mastitis in cattle and as therepeutic agents in poultry and animals, including man.
The compounds are especially useful in the treat-ment, particularly by oral administration, of infectious diseases caused by many Gram-positive and Gram-negative bacteria. Also included within the invention are easily cleavable esters of the above acids and pharmaceutically acceptable acid addition salts of said esters.
Description
~6Z70~L
~e In on~
(1) ~ , - The cephal-osporins of the present invention possesg the u~ual ~ttr~butes o~ such compounds and ~re . .
particularly useful in the treatment of bacterial infections by oral adminl~ration.
~e In on~
(1) ~ , - The cephal-osporins of the present invention possesg the u~ual ~ttr~butes o~ such compounds and ~re . .
particularly useful in the treatment of bacterial infections by oral adminl~ration.
(2) Description of the_prior art.
A. Ce ~ in~ in Gener~l Cephalothin and cephalorldine are well-known ~ntibacterial agent~; see U.S. patents 3~218,318;
A. Ce ~ in~ in Gener~l Cephalothin and cephalorldine are well-known ~ntibacterial agent~; see U.S. patents 3~218,318;
3~449,338 and 3l498,979. The litersture ~lso coneain~ considerable d~a on the activlty of ~ephaloglycin and cephalexin; ~ee U.S. patents 3,303~193~ 3,507,861 and 3,560,489 and Grea~
Britsin 9859747 and 1,054,806. .Newer cephalo~-porins include cefa~olin ~nd cephapirin; see U.S.
p~tent 3~516,997 ~and also Netherlands 68/~5179 ~F~rmdoc 34,328) ~nd South ~Africa 68/4513]~ and .S~ paten~ 3,422, lOOo The literature on cephalosporins he~ ~een revie~ed by E.P. Abraham, Quart, Rev. ~London3 ~1, 231 ~1967) by E. Van Heyningen, Ad~an. Drug Res., 49 1-70 (1967) and briefly in Annual Report~
in Medlcinal Chemis~ry, Academ~c Press, Inc., lll Fifth Avenue9 New York, New York, 10003, by L. C. Cheney on pflges 96 and 97 (1967) and by K. Gerzon and R. B. Morin on pages 90~-93 (1968~
~nd by Gerzon on p~ges 79-80 (1969~ and by L. H.
Gonover on page3 101-102 (1970). New cepha~o -porin~ are frequently reported at the annu~l Interscience Conference on Ant~microbial Agent8 ~nd Chemotherapy ~8 illustrated by Sa~siver et al., An~im-~crobial Agents and Chemotherapy -1968, American Society ~or Microbiology, Bethe8d~, -.
M~ryland, p~ges 101~114 ~19~69~ and by Nishi~a ~ .
.. . . . . . . .... . _ . .. .... : . . . _ _. _... . _ . _ . . . . . : . . . _ . . .. . ..
~,6'27~
et ,~1 ., ibid ,, 236^243 (1970) . rwo excellent rev~ew,~3 are The Cepha losporin,~s; Microb~ologic,al Shemical ,and Pharmacological Properties and U,3e iTI Chemotherapy of Infection, L. We~nste~n ~nd R. ~apl~n, Annals of Internal Medicine, 723 729-739 (1970) and Structure Ac~civity Relat~on-shlps A~ong Semisynthetic Cephalosporin,3, M.L.
Sas"~ver ~nd A. Lewi,s" ,Advances in Applled Micro-b~ol~y, edited by D. ~erlman, 13, 163-236 (1970), Ac~d~ c Press, New York. Two more recen~ review,~3 ,~re ~ Lactam Antibiotlcs: Thelr Physicochemic~l P~oper~ies ~nd Biological Activi~ie3 in E~ela'cion to Structure, J.P. Hou ,and J.W. Poole, J.
Ph~rmaceutical Sciences, ~4~, 503-532 (April9 ..
1971) and Chemistry o~ Cephalosporin Antibiotics, R. ~. Morin and B. G. Jackson, Fortschr. Chem.
Org. Naturst, 28, 343-403 (1970) which ~ncludes a "~eetlon on nucleophilic di.splacement of the 3ce~te group ,~t pages 370-373.
The preparation of various 7- [a-amino-arylacetamidol- cephalosporanic acids and the eorre~ponding desacetoxy compounds in ~hich aryl represents unsubs~ituted or substituted phenyl or 2- or 3-thienyl is described, for example9 in ~ritish Specif~cations 98S,7479 1,0173624, 1,054,806 and 1,1239333, in Belg~um patent 6969026 (Farmdoc NoO 29,494)~ in U.S. patents 3,311~6~1, 3,35~8589 3,48g,750, 3S48O,751, 3,489~752~ 3,513,260 ,and 3,575,969, in J~panese patent 16871/66 (Farmdoc 23,231), by Spencer et ,31., J. Med. Chem.~ ~ , 746-750 (1966), by Ryan et al., J. Med Chem., 12, 310-313 (1969) ,~nd by Kurita et ,al., J. Antibiotic,s (Tokyo) ~A) ': ':'' ' ~6~27~L
19, 243-249 (19S6) and see also U.S. patent 3,485,819. Briti~h Specification 19073,530 includes a disclosure of the preparation of ~uch compounds by acyla~cion of silylated 7-ACA.
Netherl~rlds patents 68111676 (Farmdoc 36,349) and 68/12382 ~Fanndoc 36,496) and U.S.
patents 3,489,750 ~nd 31489,751 disclose ring-substituted cephaloglycins .
B . 3 -Thiomethylc epha losporins V~rious cephalosporins, including cephal-osporLn C on occasion but not cephaloglycin, have been reacted with nucleophilic? aromatic mercapt~n~ to produce compounds having the ~truoture Acyl ~ C~ 2 O--C l~ C - CH2 - S - Ar \~
..
COOH.
In U~S. p~t~nt 3~278"531 Ar ~ phenyl or certsin 8ub8tituted phenyl8 or certain aro~tic hetero-cyclie rings named, ~or example, in column 5.
Siniilar nucleoph~les~ e.g, 2-mercaptopyrimidines, ~re di8closed in ll.S. 3,261,832 and Great Brita~n 1,101,422 and U.S. 3,479,350 and U.S.~ 39502,665J!
all issued to Glaxo. A parallel disclo~ure i~
found.in Great Britaill 1,109,525 to Ciba, ~,g, in de~inition "hl1 for R3. Additional nucleo phll~s o~ this type were disclosed by~ Fu~isawa in Belglum 7149518 (Farmdoc 35,307; Netherl~nds _4_ Z7C~L
68/06129 ~nd South Africa 2695/68), in Canada 818,501 (Farmdoc 38,845), in Great Br~tain 1,187,323 (Farmdoc 31,936; Netherlands 67,14888), in U.S. 3,530,123 and in U.S. 3,516,997 (Farmdoc 34,328; Ne~herlands 68/û5179) which includes the compound named cefaz~lin, which has ~ tetrazoly-l~cetyl s~decha~n on the 7-amino group and a 5-methyl-thladlazolylthiomethyl group at the 3-position and is described at ~ome length ln the scientiflc literature, e.g. in Antimicrobial Agent~. and Chemotherapy - 1969, American Society for Mlcrobiology, Bethesda, M~ryland ~ pages 236~243 ~nd in JO Antibiotics (Japan) 23(3), 131-1~8 (1970).
Replacement of the 3-,acetoxy group of a cephalosporin by various heterocyclic thiol.~ has been disclosed in U.S. 3,563,983 an~ in Netherlands 70105slg ~Farmdoc 80,188R) where the ~idçchains were, for ex~mple, 7-a-amlnophenylacetamido and ~ypical heterocyclic thiols were 2-methyl-1,3,4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5- chiol . ~
Various cephalvsporins having ~hP structure , ~
/S\
Acyl - - NH - CH ~:H ~H2 o~ I t ~ CH2 - S - a lkyl - ~C : ' ' COO~
:
~n which acyl represents various sldechaine including a-aminophenylacetyl have been d0scribed ,. .. , . ..... . .. . . . .. _ . . . . _ . .. .. _ . . . .. . . .
~0627(~
in some of the a~ove ~nd by Glaxo in ~elgium 734,532 (Farmdoc 419619) and in Belgium 734,533 (Farmdoc 41,620) ~nd by Lilly in Belgium 743,754 (Farmdoc 41,150R).
Cephalosporins hav~ng the structure Acy~ - NH - CH ~ CH2 ~ ~ =L _ N ~ - CH2- X
- \C~
.
COOH
S N -'. Il 11' ~
where X ~nclude~ - S - C - ~3nd - S - C - sre disclosed in m~ny patents including some of ~he above and in U~S. `3,239,516, 3,239,515, 3,243,435 9 3,25~,4~, 3,4319259, 3,4469~03, 3,278,53~ -3~261,83~ and 3~573,298.
: Related p~blications ln ~he scient~fic .
l~terat~re include J. Med. Chem. 8, 174-181 (1965) : and Ji~Chem. Soc. ~London~ 1595-1605 (1965), 5015-5031 (1965) and 195~-1963 (1967).
: C. 3-AcylthiomethylcePhalospor~n~ - -~ The following publication~ and patents dis=
: close certain ~dditional 7-ACA derivatives containing a 3-acylthior,;ethyl moiety (in which phenyl is abbrevlated as Ph): ~
l.~:G. F. H~ Green, J. E. Page, ~nd S. E.
: ~ Stanifo~th, J. Ghem7 Soc., 1545-605 (1965).
Thi~ re~erence gives the proton ma~netic re~on-~nce spectra of the 3-benzoylthiomethyl d~er~va- `.
tive o~cephalothin.
Co~ker et ~1., J. Chem. Soc.~ 1142-1151 :`
(19S6) ~dds thiopicolinyl ~nd references Belgium 650,444 .
. .
1~6Z7~
.
2. J. D. Cocker, et 81. 9 J. Chem. Soc.,5015-31 (1965) discloses compound having the structure ~NoF~
`1 ~CH2- S - C ~
QûH
wherein R ha~ the following meanings: PhCH2-, CH2-, CH3-S-C~12-, Ph-CO-S-C~-, HOO~ (CH~ 3-3. Glaxo~s U.S. 3,261,8'32 disoloses compounds havin~ the ~truc~ure - :
. . _ .
~SllC112CON~ S~1 N~f ~`CH~ -Y : ' COOH
wherein Y ha~, ~or example, the follo~ing mean-lngs ~
'5~~ s-co~3 s-co ~here R is C1130-, ~N02, -CN~ CH3S~
_ 7 _ -s~ s--~o~
5 ~o~ S-CO-CH2~, -s-co-(cH2)3cH3 Equ~valents are Netherlands 64/08066 (Farmdoc 15534) and Great Britain l~:L01,424.
4O Glaxo'~ Netherlands 65/0S818 ~Farm~o~
1~306) discloses the react~Lon H2N~S
J=N~cl~2-s~cl)-ph ~ CH2COCl : :
COOH ~ ~
:, .
~
~ N \ GH2-S-CO-Ph AT1 ~quiv~ler~ U, S . 3 " 502, 665 .
' ~ ., : ~ /
~ ~ ~2 7~ ~
5. Gl~xo's Netherlands 64/11521 ~Chem. Abstr. 9 63: 13,281d (196S)] discloses the reac~ion ~ep~3 C ~in~5i~L~ HOOC-CH (CH2)3CONH ~ S ~
NH2 ~ ~ N ~ CH2-y COOM
whereln Y -S-CO-Ph or -S-CO ~
Equ~valent~ are Great Britain 1,101,422 and Can&da 796,747 (Farmdoc 17362).
6. Cib~'~ U.S. 3,555,017 disclo~es compounds having the structure . R
: R / C~-CO-NH ~ ~
COOH ~;
Rl ~nd R2 ~ halogen. ;- :
As usual in Ciba 18 patents directed prim~rily to novel ~id~ch~ins ~t the 3-posltion, the R group ~bove i~ defined broadly ~s the residue o~ ~
c~rboxylic acid and m~y be illustrated by phenyl, the residue of thi~benzolc acid. Equlv~lents ~re Belgium 7O~D24~ rmdoc 33~276), Great Britain 1,211,747 ~nd French 19575~554.
O g O
.... . ... . . .. .. . .. .. . . .. . .
~L~627(~
7. Ciba 's British 1,211~718 discloses com-pounds havlng the structure r O ~ C ~S .
R ~s in 6 above.~quivalents are Belgium 708,311 (Farmdoc 339277) and U.S. 39557,104. ::
8. Ciba 's Belgium 7519526 ~Farmdoc 90,178R) d~ sclo~es compounds having ~the s~ructure N3~ H2C-NH~ S ~
SC-R
COO~I ' ':
R a~: ~in~ 6 above.
An equ~valent i~ Netherland~ 70/08237.
:~ : 9. Ciba's South Africa 69l8436 di3closes : -. : compound~ having the structure `, ~N ~U ~C~ ~/ ~ O ". ~.
N ~ ~ S~
COOH
R a~ in 6 above.
.
E~ivalents ase Belgium 743,014 (Farmdoc 43,126RS
and ~etherlan~ds 69l18611.
.
~OÇ;Z7~ :
10. Ciba's ~outh Africa 69/8399 disc1ose8 compounds having the ~tructure N~; N~C \ CO-NH
~,==1 '~ H2 COOH
R a~ in 6 above.
Equlva1erlt~ are Bel~ium 742,933 ~rmdoc 41,568R) ~nd Netherlands 69/18531.
11. Giba's South Africa 68/8185 disc1oses compounds having the struc~ure H2 ' 0~ H2 R as ~n 6 above, COOH
An equivalent ~s Netherland~ 68118868.
12 . Ciba ' s Netherlands 68/18868 d~ sclose~
compounds having the structure _~ H2 : ' .
N ~ V~ O-NH
N~ ~ SC.-R
OOH
.
R a5 in 6 abov~. ~
Equivalents are South Africa 8120t68~ German 1,817,121 and Belg,lum 726~316.:
: ': -... ....... .. . . . . ..... .. . .. . .. .. : . . .. . .
~(~6~7(~1 13, Fu~isawal~ Great Brital~ 19187,323 for example, ~ page 5, lines 67-71 disclose~
compounds having the structure - ..
~ O
o\/ ~ Z ~ ~ S ~
R / R O / ~ / SC-R
, COO~
in which R represents ~ethyl, thienyl, pyridyl9 etc. ~nd wherein general disclosure is m~de of other heterocycllc groups a~: at pages 1 and 2.
Equivalents are Netherlands 67/14888 (Farmdoc 31,936) and U.S. 3,530~123.
.
~ 14. Fu~isawa~ Belgium 714,518 (Farmdoc 359307) discloses ~among many others) compounds having:the ~tructure R . :.
/ ~ B ~s 1l C-R
~ ~,CO~
w~ereln~ is as 13 above.
Equivalents are Netherlands:68/06129 and South~African 2695/68.
, ...
:: .
~ - 12 - ~
.. ... . . . , . _ . _ , . .. . _ . . ., . . _ . .
3L[36;27ai1 15 . Gl~xo ' s U . S . 3, 243 ,435 and Belgium 650,444 (Farmdoc 15,535) disclose generally a va~t variety of compounds h~ving the structure Acyl-NU-CO-llH~ CH~S
COOH
wherei.n ~, is defined, e.g. in column~ 1 and 4, t~ ~nclude various heterocycllc groups.
16. Cibals South Africa 65/6950 (Farmdoc 22 ~192) discloses compounds having ~he struc~cure N~ ~ C~lR2-~-N~S
,~N )~ SC-R
.` ~00~
in ~ich R in Example 20 is phenyl.
Equivalen~s ~re Great Br~tain 1,109,,525 ~nd Canada 807 ,651.
~ 17 . Glaxo ' s U. S ~ 3 ~47g , 350 d~sclo~os ~ proces~
for producing 3-pyridiniuImnethyl cephalosporin~
which utilizes ~s ~n intermediate compounds of the type~deser~bed in rcference~ 2 and 3 above~
' :
.
13 ~
~62710~L
18. Ciba 's U S. patent 3,7579013 d~scloses compounds of the formula ' 2~ ~\~ CH2-S-C-R
COO
wherein R3 i~ (lower3 alkyl and R is as in 6 above.
~ Lg. Bristol9s Belgi~n patent 795,811 disoloses compounds of the general fonn~lla Il ,S O
Ar- l~-C~ ~ rr NH2 `o~l N ~CH2-S-C-Z
COQR
wherein ~Ar is phenyl" 2-th~enyl or 3-thienyl; R
~ : : ls hydrogen, pivaloyloxymethyl, acetoxyme~chyl, ; methoxyme~hyl, acetonyl or phenacyl; and Z is ~.
H3 , _ N
~CU3 ~
~: 3-i~oth~zolyl 9 4-isothiazolyl or 5-isothiazolyl .
:
- 14 - ~
..
. . . ... .. ............. ., .. .. . .... . . . . . .. . . ....... ... , . . . .. ~ .
~9627~
This specification comprises the novel compounds having the D-configuration in the 7-sidechain and the formula ~r-CH-C-NH
~H2 ~ ~ ~ C~2~
COOR ::
wherein Ar is phenyl, 2-thienyl or 3-thienyl; R is hydrogen, :~ -pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl; and R' is 2-thiazolyl or 5-methyl-2-thiazolyl or -~
3-methyl-1,2,5-oxadiazol-4-yl, and their pharmaceutically acceptable salts. ;~
Summary of the Invention In a broad aspect the present invention provides a process for the preparation of a compound having the D- .
configuration in the 7-sidechain and having the formula `
H-I-N~ ~ S ~ 9 (I) H2 ~ N ~ C~ C-R' eOOR
: ~ .
whereLn R is hydrogen, pivaloyloxymethyl, a~etoxymethyl, : met oxymethylp acetonyl or phenacyl; and R' is 2-thiazolyl or 5-methyl-2-thiazolyl or 3-methyl-1,2,5-oxadiazol-4-yl; or a `
pharmaceutically acceptable salt thereof, comprising:
~a1 when the starting material includes the desired final substituent in the 7 position of the cephem nucleus:
reacting a compound of the formula `--Q ~ .
~ fH-C-NH 3 NH2 ~ ~ CH~OCOCH3 C2~ :
15 - ~
.. .. . . . ~ ~.. . . ~ .. . .
~(~36Z 17~L
or an easily cleavable ester or salt ~hereof wi.th a hetero-aromatic thiolcarboxylic acid of the formula o R'-C-SH
wherein R' is as defined above or a salt thereof to form the desired compound of formula I, or , ' (b) when the starting material includes the desired ~:~
final substituent in the 3-position of the cephem nucleus~
10 reacting a compound of the formula , ' ~2N ~ S ~
O N ~ CH~S-C-R' CO2~ .
:
wherein R' is as,defined above or an leasily cleavable ester or .~
salt thereof with acid having the formula '. ' .. ~... ...
CH-COOH
I
NHB ' '' or an acylating derivative thereof, wherein B is an Emino-protecting group and removing said amino-protecting group B to : :
produce the desired compound of formula I or an easily cleavable ester or,pharmaceutically acceptable salt thereof; -' . -':
and, if dPsired, in the case of (b) either before or after ~, removal of protecting group B ~i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or ,.
(ii) converting the product in the form of an easily cleavable ~ -'15(a) ~
. - . ... .
:
. .:
7~
ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
The pharmaceutically acceptable salts referred to above include the nontoxic carboxylic acid salts when R is hydrogen, e.g. nontoxic metallic salts such as sodium, potassium, calcium and aluminum, the ammonium salt and salts with nontoxic amines, e.g. trialkylamines, procaine, dibenzyl-amine, N-benzyl-pLphenethylamine, l-ephenamine, N,N'-dibenzyl-ethylenediaminel N-alkylpiperidine and other amines which have been used to form salts of penicillins. Also included within the definition of pharmaceutically acceptable salts are the nontoxic acid addition salts (amine salts) of the acids and ~
esters of Formula Ia, e.g. salts with mineral acids such as hydrochloric, hydrobromic, hydroiodic, phosphoric and sulfuric and salts wi~h organic acids such as maleic, acetic, citric, oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic and malic.
: . ...:
- 15(b) -.. ~, . .
1~627~
Preferred c~pounds of the present lnvention ~re the eompounds h~ving ~he D-configuratlon ln t:he 7~idech~in ~nd ~he formula Ax ~CH-C-NH
N~2 ~ c~2_s_~
COOR
Ib wherein Ar i6 phenyl, 2-thieriyl or 3-thienyl ~nd R iz hydrogen, p~v~loyloxymlethyl~ acetoxymethyl, ~ethoxyme~chyl s acetonyl or Iphenacyl; and the ph~rmaeeutically aeceptable salts thereof.
Preferred compounds of formula Ib are those in which ~9r i~ phenyl. An e~plecially preferred ~ompound of formula Ib ~ tElat in which Ar i8 ph~nyl ~nd R i9 hy~rogen; or the sodium or potas~ium s~lt ~her~of.
More preferred compounds of the present invention are the compourlds having the D-con-figuratiorl in the 7-sldQch~in and the fo~mula Q
Ar -CN-C -NU ~l CN2 _ 5 _C ~ CU3 COOR
. Ic wherein Ar i~ phenyl, 2-th~enyl or 3-thienyl and R i~ hydrogen, pivaloylox~nethyl9 acetoxy-methyl, methoxymethyl, ~cetonyl or phenacyl 9 ~nd the ph~rmaceutic~lly ~ccept~ble salts thereo~. Preferred compound~ of formul~ Ic are tho e in whieh Ar is phenyl. An especi~lly ~L~627~
preferred compound of formula Ic is that in which Ar is phenyl and R is hydrogen; or the sodium or potassium salt thereof.
Further preferred compounds of the invention are the compounds having the D-configuration in the 7-sidechain and the formula A~ N~ H -5-~
Id wherein Ar and R have the meanings given above and the pharmaceutically acceptable salts thereof.
Preferably Ar is phenyl and R is hydrogen; or the sodium or potassium salt thereof.
Also included in this invantion ~re the co~pounds ~u~ed as sither int~ermediates or met~-bolie precursors) in ~hlch thle amino group i8 "blocked" by substituents suclh a~ t-butoxycarbonyl, c~rbobbn~yloxys~formy~ o-nitrophenylsul~enyl9 -trichloroethoxycarbcnyl~ 4-oxo-2-pentenyl-2, l-c~rb~me~hoxy-l-prspenyl-2 snd the like. P~rtic-ularly lncluded in ~uch blocking groups are the ketones (especially acetone) snd the ~ldehydea (espec~ally formaldebyde ~nd acetaldehyde) di~-closed for example ln U.S. patents 3,1989804 ~nd ~347,851 and the ~ketoesters ~nd the.~-diketones di~closed for ~x~mple ln U.S. patent ~9325,479 ~nd the ~ketoamides disclosed in Jap~n 71/24714.
The pr~ent in~ention also includes a process for the p~eparation of ~ eompound having the formula p A~_q~ ~T~S
H2 ~ ~ CH2-S-C-R' CC)~
' wherein Ar is pheny~ t~ienyl or 3-thienyl;
R' is 2-thiazolyl or 5-methyl-2-thiazolyl or 3-methyl-1,2,5-oxadiazol-4-yl; and easilY cleavable esters and pharmaceutically acceptable salts thereof;
which process comprises either ~A) reacting ~ compound o~ the formula ., M2N --- I , s ~
o~ ~CH2S-C~R' II
_._ wherein R' is as defined above or an easily cleavable es~er or salt thereof with an acyla lng deriv~tive of an acid having the for~ula :
Ar-~H-COOH
~HB .
III `
, ~ , .. .
wherein B is an amino~protecting group and Ar ~s a8 defined above and removing sa~d amino-protecting group B to produce the desired com-pound o formula I or an easily cleavable es~er or ph~rmaceut~cally acceptable s~lt thereof and, if deslred~ ei~her before or after removal o~
pro~ecting group B (a) converting by methods -known ~ se the product ~n the form of the free acid or 8alt the~eof to a corresponding e~sily cleavable es~er or pharm~ceutically ~ccept-able sàlS thereof or ~b) converting by methods known~per se the product in the ~orm of an easily cleavab1e ~ster or ~alt thereof to the correspondlng free aci~d compound or pharmaceutically acceptable s~lt thereo~; or , .
~8 -.
l~Z7~1 (B) reac~ing a compound of the formula Ar-CH-C-NH ~ ~
~ 2 0 ~ ` ~ CH2ococH3 IV
whereln Ar is 3S defined above or an easily cleavable es~er or salt thereof with a hetero- `
aromatic thiolcarboxylic acid. of the formula R'-C-SH
:~ .
where~n R' is as defined abo~e or a s~lt thereof to form a compound of formula I or an easily cleavable ester or pharmaceutically acceptable 8alt thereof and, if desired, (a) converting by methods known ~ se the product in the form of the free acid or salt thereof to ~ cor~esponding easil~ cleavable ester or pharmaceutieally accept- :
able salt thereo~ o~ (b~ converting by methods `
~nown ~ se~the product in the form of a~n easily cleavab-le ester or salt thereof to the corresponding free acid compound or pharmaceutically ~cceptable sal~c ~ thereof.
. .
The easily cleavable esters referred eo above include ester groups which are removable by methods, e.g. chemical or:enzymatic hydrolysis~ which do not result ~n any appreciable destruct~on of the remaining~-portion of the cephalosporin molecule.
~xamples of suitable esters include those disclo~ed in U.S. patents 3,284,451 ~nd 3,249,622 and U.K.
patents 1,229,453 and 1,073,530. Parti~ularly . .
- 19 - ' , ~ ~ ~2 7~ ~
preferred esters are the piv~loyloxymethyl, acetoxymethyl9 methoxymethyl, acetonyl and phen~cyl esters.
In one method of preparing the novel cephalo-~porin compounds of the present invent~on, a 3-thio-lated-7-aminocephalosporanlc acid compound of form~l~ II or an easily cleavable ester or sa1t of ~id acid or ester 18 acylated w~th ~he appro-priate acylating derivative of formula III.
The 3^thiolated-7~amlnocephalospor~nic ~cid lntermed~a~e of formul~ II may be prepared by - di~placement of the 3-ace~oxy group of 7-amino-cephalosporanic acld or a salt ~hereof wi~h the appropriate heteroaromatic thiolcarboxylic ~cid or g salt thereof. The displacement of an ester group with a thiol group is a known reaction and i~ preferably accomplished iLn aqueous solution at a temper~ure oi ~t. least room temperature and preferably within the range of about 50 to 100C.
. iLn the presence of ~ mild base such as sodium b~carbon~te.
Th~ claimed compounds may then be vbta~ned ; by a~ylatlo~ ~ccording ~o known methods of the 7-amino group o~ intermediate II with the acyl3ting agent of`formula I~I.
: Because of the low solub~lity of the compound~
of formNla II in common aqueous an~ non-aqueous : 801~ent~, lntermediate II ls preferably converted prior to:the acylation reac~ion ~o an`easily cte3va~1e~ester or ~cid~addi~ion salt thereofo The procedures for preparing such es~ers are d~sclo~ed in the literature ~nd are well-known t~ those skilled in the art of penicillin and cephalosporin chemistry. One preferred me~hod espec~ally useful for preparing 'ch~ most preferred e~sily hydrolyzed esters, i.e. the piv~oylox.ymethyl, - ~0 - ;
, . .. .. . . _ . . _ .. . , _. . . . . . ..
1~6Z~7~1 acetoxymethyl 3 methoxymethyl, acetonyl and phen-acyl esters, is disclosed in U.S. patent 3,284,451 This reference descrlbes the esterification of sodium.cephalothin with the appropriate active chloro or bromo compound (e.g. phenacyl bromide, ~hloroacetone, chloromethyl methyl ether, pivaloyloxy;
methyl chloride, acetoxymethyl chloride3 followed by enzymat~c removal o~ the thienylacetic acid ~ide-ch~in. In another good method ~he ~rie~hylamine salt of 7-aminocephalosporanic ~c~d i~ reacted directly with the active halogen compound as ln U.K. patent 1,229,453. The compound of formNls II n~y also be converted to ~ ~ilyl ester as by the methods described ln the literature, e~g. .
U.S. patent 3,249,622. The silyl ester group may be removed following the acylation reaction : .
by hydrolysis or alcoholysls. ~ -Prior to the ~cyla~ion reaction the amino group o~ acylating agent III is protected by a conveneion~l amino-protect-img group B which may be read~:ly re~oved ~t the conolusion of the react~on. Examples of suitable amino-protect~ng groups include tbutoxycarbonyl, carbobenzyloxy, 2-hydroxy-l naphthcarbonyl, trichlor~ethoxyc~rbonyl, 2-ethoxycarbonyl-1-methylvinyl and 2-methoxycarbonyl-l-methylvinyl. A p~rticularly valuable ~locking group~is;a prOtOII, as in the compound of the formNls .
Ar-~H-COCl wherein Ar i~ phenyl, 2-thienyl or 3 thienylO The preferred amino-protecting groups are t-butoxy-carbonyl, the proton and a ~-diketone or a ~-keto~ -ester as in U.K. paten~ 1,123p333 or U.S. patents .
.. ... . .. ... . . .. . , . . . . _ .. . . . . . .
~L062~
3,325,479 and 3,316,247, e.g. methyl acetoacetate9 or a ~^ketoamide as in Japan 71/24714. When the t-butoxycarbonyl~ ~-ketoester, ~ diketone or ~-ketoamide protecting groups are employed~ it ls preferred to convert the acylating acid con-taining the blocked amino group to a mixed nnhydride~ e.g. with ethyl or isobutyl chloro-formate, before reaction with compound II or an e~ter or ~alt thereof. After the acylàtion coupling reaction, the amino-protecting group B
m~y be r~moved by methods known ~ se to form the desired product of formula I. Thus, for example, the t-butoxycarbonyl group m~y be removed by use of formic acid, the carbobenzyloxy group by catalytic hydrogenation, the 2-hydroxy-1-naphth-carbonyl~group by acid hydrolysis, the trichloro-ethoxycarbonyl group by tre~stment with zinc dust ln glacial acetic acid, ~he proton by neutrali-zation, etc. Obviously other func~ion~lly equ~v~
alen~ blocking groups for an amino group can be used and such groups are considered within the scope of this invention.
Acylation of a 7-~mino group of a cephal-~sporin ~ 8 a well-known reaction and ~ny o~ the func~ional equivalents of formula III commonly used ~s acylating agents ~or primary amino group~
m~y be employed. Example~ o sui~able acylating derivatives~of the free acld include the co~re5-ponding ~cid anhydride~, mixed anhydr~des, e~g.
alkoxyformic anhydrides, ~cid halides, acid az~des~
~ctive esters an~ active ~hioesters~ The free acid m~y be coupled wi~h compound II ~fter first reacting said free acid with N,N1-dimet~ylchloro-~ormimin~um chlor~de or by the use of enzyme6 ~r .
'~ ' ''' .
, . . .. . ... . . . . .. . ... . . . . . . .. .. . .. . .. . . . .
~ 0 6Z 7 of ~n N,N'~carbony1diimidazole or an N,N' c~rbonyl~
ditr~azo1e or a c~rbodiimide reagent, e.g. N,N!-diisopropylcarbodiimide. N9N'-dicyclohexylc~rbodi-imide or N-cyclohexyl-NI ~2-morpholinoethyl)~
c~rbodiimide or of alky1ylamine reagent or of an i~oxa~olium salt re~gent. Another equivale~ of ~he free ac~d i~ ~ corre~ponding azollde, i.e., an amide of the corresponding acid whose a~ide nltrogen i~ a member of ~n quasiaromatic five membered ring containing ~t least two nitrogen ~toms, ~.eO 3 i~idazole, pyrazole, ~he triazoles, benzimidazole, benzotri~zole and their ~ub~tituted deriv~t$ves. Another react~ve derivat~ve of the pheny:lglycine acid of formNla III i8 thè N~carboxy anhydride (Leuch'~ anhydrid~) . In this structure ~he group which activfltes the earbo~cyl group also serves to protect the ~m~no group. A particularly preferred acylating agent i8 the acid chloride hydrochloride of the iEormula Ar-~H-eOCl NH2 ~ HCl ,.
whioh also ~erves ~ du~l function of osrboxyl ~ctivation and amino proteotion. Mention was m~de above of the use of enzymes to couple the free ~cid with its blocked amino group wi~h c~m-pouFad II. Included in the scope of 6ueh proce~ses ~re ~he use of an ester, e.g. the methyl ester, of that free acid with enzyme~ provided by v~riolls microorg~nisms, e.g. those described by T. Takahashi et al., J~ Amer. Chem. Soc., 94(11~ 9 4035-4037 (1972 ~nd by T. Nara e~ ~1., J. A~tibiotics (Japan), 24(5), 321-323 tl471) and ln West Germ~ny 292169113.
:: - , . .. , .. ,. . , .. , . ~ , .
~ 0 6~ ~0 ~
The particular proce~s conditions, e.g.
tempersture~ solvent, react~on time, e~c. ~elec~ed for the acylatlon coupling reactlon are determined by the nature of the acylation method used and ~re known to those skilled in the art. Generally it i~ useful to add an organic tertiary amine, e.g. tr~ethyLamine, N~N-dimethylsniline, ethyl-piperidine, 2,6-lutidine or quinoline, to ~erve as a proton acceptor or salt-forming agent. A
preferred method ~llustrated in the examples which follow involves formation of a s`ilyl ester, e.g. with trimethylchlorosilane, of the inter-mediate of formula II and acylation of this sily~
l~ted intermediate ~n dry methylene chloride at temperature of below room temperature ~nd prefer-ably about 0C. with the appropriate chloride hydrochloride acylating age!nt of formula III in the presence of ~n organic tertiary am~ne.
At ~he concluæion of the acylation reaction, the acylated intermediate is sub~ected to aqueous hydrolys~s to provide the desired cephalo~porin produo t .
~ The eompounds of the present invention m~g be isolated in any of the ways ~ustomarily empIoyed for ~he isolation of similar cephalos-porin~. Thus 9 the product may be ob~ained ~ -~he neutral moleeule~ although thi~ is prvbably more~accurately represented ~s the æwitterion, or ie m~y be ~solated as a ~al~. Form~tion of the de~ired pharmaceutically aceepta~le carboxylic acid or acid addition salt ~s carried out by known methods, e.g. reaction of the ~cid with ~n appro~
pria~ base or acld. ~ ~ :
At the conclusion of the acylation reaction ~he product obt~ned m~y be converted (before or 1 0 6 2 7~ ~
after r~moval of the ~mino-proteceing group) by methods known per _ to another desired product of formula I. Thus, the compound of formwla I
in the orm of the free acid or a s~l~ thereof m~y be converted by known methods ~o a corres~
pondlng easily cleavable ester or pharm~ceutic~lly ~ccep~able s~lt thereof. Similarly, the product of formula I in the form of ~n eaælly cleavable ester or s~f~ thereof may be converted to the ree acid product or pharm~ceutically ~cceptable ~alt thereof by removal of the esterifyi~g group9 e.g~ by ~queous or enzymatic hydrolysis ~a~ wlth human or ~nimal serum~ or acidic or alk~line hydrolysls or by catalytic hydrogenation or by treatment with sodlum thiophenoxide as t~ught in U.S. patent 3,284,451.
In another method of preparing the compounds of the present invention, 7-am~nocephalosporanic ~cid or a salt thereof is acylated with the acid oiE formul~ III or an acylating deriv~tive thereof to fo~m ~ 7-acylated cephalosporin compound of the formula ~ .
Ar-lH_ 3 ~H - -- f ~
NH2 ~ ~ CH20COCH3 CO2~' , ., . I~
Compound IV in the form of the free acid or an easily cleav~ble ester or salt thereof is then reacted ~ccording to the process of the present invention with ~ heterocyclic thiolc~rboxylic acid of formul~
V or a salt thereo~ mos~ preferably the sodium ~L~6Z7~1 or potassium salt. The displacement resction i8 preferably conducted in aqueous solution ~t temperatures of about 50CO or higher in an inert atmosphere, e.g. under nitrogen. The product of the displacement reaction m~y, if desired~ be converted to a pharmaceutically acceptable salt by treatment with an appropr~ate acld or base. As in the ca~e of the alternate process described above for prepara~ion of the compounds of formNla I, the product in the form of the free acid or salt thereof may be converted to a corresponding easily cleavable ester or pharnaceutically acceptable salt thereof or, alternatively, ~he product in the form of an easi1y cleavable ester or salt thereof may be .
converted to the free acid or pharmaceu~ically scceptable salt thereof.
The easily cleavable esters of the compound of formula I are use~ul as i.ntermediate8 in the produc~lon of the free ~cid product. The piv~l-oyloxymethyl 9 acetoxymethyl and methoxymethyl esters are al~o useful as active antibac~erial agent~s 6ince on oral administration they are rapldly~hydrolyzed ~o the active me~aboli~e.
These three esters are of particular ~nterest because~ they provide on oral administration different ra~es and amounts of ab~orption and give differing concentrations of ~he active anti-bacterial agent in blood and tissues.
The preferred and most active compounds of ' the presen~ invention ~re those having the D-configuration at the a-carbon atom in the 7-side- l chain, that is, those m~de from D~ 2-phenyl-~
glycine, which is also called D-~ a-amin~phenyl- .
acet~c ac~d,.and D-(-)-2-thienylglycine and D-(-~-~062~C)1 3-thienylglycine. In addition, the conf~gura-tion at the two optically active asymmetric centers in the ~-lactam nucleus is that found in cephalospor~n C produced by fermentation and in ~he 7-aminocephalosporanic acid derived there-from.
The pharm~ceutically active compounds of the present invention are potent antibacterial :
~gents useful in the treatment of infertious dlseases in poultry ~nd animals, including man, caused by m~ny Gram-posi~ive and Gram-negative bacteria. The active compounds are also of value ~s nutritional suppllements in animal feeds .
and as agen~s ~or the trealtment of mastitis in ca~le. The preferred compounds have also been unexpectedly found to be efficiently absorbed uposl oral administration.
Despite the synthesis of a large number of cephalosporins reported in the scientific and patent liter~ture, the only cephalosporin at th~
present ~me having suffic;Lent actlv~ty ~nd or~l ~bsorption for general: oral use is cephalexin.
In our search for new orally active cephfllosporins, we ha~ve found ~hat the compounds claimed~ in the present ln~ention fiurprlsingly and unexpectedly possess:~good or~l absorption and at ~he~lsame ~ime appear to be superior to cephalexin ~n the~r activity against certain impor~ant pathogen~c org~ni~ms. Thi combinat~on of good oral absorp-tion~and high antibacterlal ace~vity~ln a~ceph-~losporin while highly desirable is quite uncommon in compounds reported to date. M~king~any reliable predict$on as to artivlties or oral ~bsorptions of new cephalosporin compounds from a~ examLnation of .
~62~
the propert~es of structurally analogous prlor ~rt compound~ has also b~en found to be essen-tially impossible. To illustrate the unpredic-t~bill~y of oral absorption and antibacterial activity in ~ closely related series o~ cephal-o~po~in compounds 9 Tables 1-8 below compare in vitro activitie~ ~nd oral absorption as tes~ed in the mouse of the compoun(l~ of the formula H_~_N~ S
NH2 ~ CH2-S-C-Z
D~
wherein Compound No.
.. . .
æ ~ 2-thlazolyl 567
Britsin 9859747 and 1,054,806. .Newer cephalo~-porins include cefa~olin ~nd cephapirin; see U.S.
p~tent 3~516,997 ~and also Netherlands 68/~5179 ~F~rmdoc 34,328) ~nd South ~Africa 68/4513]~ and .S~ paten~ 3,422, lOOo The literature on cephalosporins he~ ~een revie~ed by E.P. Abraham, Quart, Rev. ~London3 ~1, 231 ~1967) by E. Van Heyningen, Ad~an. Drug Res., 49 1-70 (1967) and briefly in Annual Report~
in Medlcinal Chemis~ry, Academ~c Press, Inc., lll Fifth Avenue9 New York, New York, 10003, by L. C. Cheney on pflges 96 and 97 (1967) and by K. Gerzon and R. B. Morin on pages 90~-93 (1968~
~nd by Gerzon on p~ges 79-80 (1969~ and by L. H.
Gonover on page3 101-102 (1970). New cepha~o -porin~ are frequently reported at the annu~l Interscience Conference on Ant~microbial Agent8 ~nd Chemotherapy ~8 illustrated by Sa~siver et al., An~im-~crobial Agents and Chemotherapy -1968, American Society ~or Microbiology, Bethe8d~, -.
M~ryland, p~ges 101~114 ~19~69~ and by Nishi~a ~ .
.. . . . . . . .... . _ . .. .... : . . . _ _. _... . _ . _ . . . . . : . . . _ . . .. . ..
~,6'27~
et ,~1 ., ibid ,, 236^243 (1970) . rwo excellent rev~ew,~3 are The Cepha losporin,~s; Microb~ologic,al Shemical ,and Pharmacological Properties and U,3e iTI Chemotherapy of Infection, L. We~nste~n ~nd R. ~apl~n, Annals of Internal Medicine, 723 729-739 (1970) and Structure Ac~civity Relat~on-shlps A~ong Semisynthetic Cephalosporin,3, M.L.
Sas"~ver ~nd A. Lewi,s" ,Advances in Applled Micro-b~ol~y, edited by D. ~erlman, 13, 163-236 (1970), Ac~d~ c Press, New York. Two more recen~ review,~3 ,~re ~ Lactam Antibiotlcs: Thelr Physicochemic~l P~oper~ies ~nd Biological Activi~ie3 in E~ela'cion to Structure, J.P. Hou ,and J.W. Poole, J.
Ph~rmaceutical Sciences, ~4~, 503-532 (April9 ..
1971) and Chemistry o~ Cephalosporin Antibiotics, R. ~. Morin and B. G. Jackson, Fortschr. Chem.
Org. Naturst, 28, 343-403 (1970) which ~ncludes a "~eetlon on nucleophilic di.splacement of the 3ce~te group ,~t pages 370-373.
The preparation of various 7- [a-amino-arylacetamidol- cephalosporanic acids and the eorre~ponding desacetoxy compounds in ~hich aryl represents unsubs~ituted or substituted phenyl or 2- or 3-thienyl is described, for example9 in ~ritish Specif~cations 98S,7479 1,0173624, 1,054,806 and 1,1239333, in Belg~um patent 6969026 (Farmdoc NoO 29,494)~ in U.S. patents 3,311~6~1, 3,35~8589 3,48g,750, 3S48O,751, 3,489~752~ 3,513,260 ,and 3,575,969, in J~panese patent 16871/66 (Farmdoc 23,231), by Spencer et ,31., J. Med. Chem.~ ~ , 746-750 (1966), by Ryan et al., J. Med Chem., 12, 310-313 (1969) ,~nd by Kurita et ,al., J. Antibiotic,s (Tokyo) ~A) ': ':'' ' ~6~27~L
19, 243-249 (19S6) and see also U.S. patent 3,485,819. Briti~h Specification 19073,530 includes a disclosure of the preparation of ~uch compounds by acyla~cion of silylated 7-ACA.
Netherl~rlds patents 68111676 (Farmdoc 36,349) and 68/12382 ~Fanndoc 36,496) and U.S.
patents 3,489,750 ~nd 31489,751 disclose ring-substituted cephaloglycins .
B . 3 -Thiomethylc epha losporins V~rious cephalosporins, including cephal-osporLn C on occasion but not cephaloglycin, have been reacted with nucleophilic? aromatic mercapt~n~ to produce compounds having the ~truoture Acyl ~ C~ 2 O--C l~ C - CH2 - S - Ar \~
..
COOH.
In U~S. p~t~nt 3~278"531 Ar ~ phenyl or certsin 8ub8tituted phenyl8 or certain aro~tic hetero-cyclie rings named, ~or example, in column 5.
Siniilar nucleoph~les~ e.g, 2-mercaptopyrimidines, ~re di8closed in ll.S. 3,261,832 and Great Brita~n 1,101,422 and U.S. 3,479,350 and U.S.~ 39502,665J!
all issued to Glaxo. A parallel disclo~ure i~
found.in Great Britaill 1,109,525 to Ciba, ~,g, in de~inition "hl1 for R3. Additional nucleo phll~s o~ this type were disclosed by~ Fu~isawa in Belglum 7149518 (Farmdoc 35,307; Netherl~nds _4_ Z7C~L
68/06129 ~nd South Africa 2695/68), in Canada 818,501 (Farmdoc 38,845), in Great Br~tain 1,187,323 (Farmdoc 31,936; Netherlands 67,14888), in U.S. 3,530,123 and in U.S. 3,516,997 (Farmdoc 34,328; Ne~herlands 68/û5179) which includes the compound named cefaz~lin, which has ~ tetrazoly-l~cetyl s~decha~n on the 7-amino group and a 5-methyl-thladlazolylthiomethyl group at the 3-position and is described at ~ome length ln the scientiflc literature, e.g. in Antimicrobial Agent~. and Chemotherapy - 1969, American Society for Mlcrobiology, Bethesda, M~ryland ~ pages 236~243 ~nd in JO Antibiotics (Japan) 23(3), 131-1~8 (1970).
Replacement of the 3-,acetoxy group of a cephalosporin by various heterocyclic thiol.~ has been disclosed in U.S. 3,563,983 an~ in Netherlands 70105slg ~Farmdoc 80,188R) where the ~idçchains were, for ex~mple, 7-a-amlnophenylacetamido and ~ypical heterocyclic thiols were 2-methyl-1,3,4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5- chiol . ~
Various cephalvsporins having ~hP structure , ~
/S\
Acyl - - NH - CH ~:H ~H2 o~ I t ~ CH2 - S - a lkyl - ~C : ' ' COO~
:
~n which acyl represents various sldechaine including a-aminophenylacetyl have been d0scribed ,. .. , . ..... . .. . . . .. _ . . . . _ . .. .. _ . . . .. . . .
~0627(~
in some of the a~ove ~nd by Glaxo in ~elgium 734,532 (Farmdoc 419619) and in Belgium 734,533 (Farmdoc 41,620) ~nd by Lilly in Belgium 743,754 (Farmdoc 41,150R).
Cephalosporins hav~ng the structure Acy~ - NH - CH ~ CH2 ~ ~ =L _ N ~ - CH2- X
- \C~
.
COOH
S N -'. Il 11' ~
where X ~nclude~ - S - C - ~3nd - S - C - sre disclosed in m~ny patents including some of ~he above and in U~S. `3,239,516, 3,239,515, 3,243,435 9 3,25~,4~, 3,4319259, 3,4469~03, 3,278,53~ -3~261,83~ and 3~573,298.
: Related p~blications ln ~he scient~fic .
l~terat~re include J. Med. Chem. 8, 174-181 (1965) : and Ji~Chem. Soc. ~London~ 1595-1605 (1965), 5015-5031 (1965) and 195~-1963 (1967).
: C. 3-AcylthiomethylcePhalospor~n~ - -~ The following publication~ and patents dis=
: close certain ~dditional 7-ACA derivatives containing a 3-acylthior,;ethyl moiety (in which phenyl is abbrevlated as Ph): ~
l.~:G. F. H~ Green, J. E. Page, ~nd S. E.
: ~ Stanifo~th, J. Ghem7 Soc., 1545-605 (1965).
Thi~ re~erence gives the proton ma~netic re~on-~nce spectra of the 3-benzoylthiomethyl d~er~va- `.
tive o~cephalothin.
Co~ker et ~1., J. Chem. Soc.~ 1142-1151 :`
(19S6) ~dds thiopicolinyl ~nd references Belgium 650,444 .
. .
1~6Z7~
.
2. J. D. Cocker, et 81. 9 J. Chem. Soc.,5015-31 (1965) discloses compound having the structure ~NoF~
`1 ~CH2- S - C ~
QûH
wherein R ha~ the following meanings: PhCH2-, CH2-, CH3-S-C~12-, Ph-CO-S-C~-, HOO~ (CH~ 3-3. Glaxo~s U.S. 3,261,8'32 disoloses compounds havin~ the ~truc~ure - :
. . _ .
~SllC112CON~ S~1 N~f ~`CH~ -Y : ' COOH
wherein Y ha~, ~or example, the follo~ing mean-lngs ~
'5~~ s-co~3 s-co ~here R is C1130-, ~N02, -CN~ CH3S~
_ 7 _ -s~ s--~o~
5 ~o~ S-CO-CH2~, -s-co-(cH2)3cH3 Equ~valents are Netherlands 64/08066 (Farmdoc 15534) and Great Britain l~:L01,424.
4O Glaxo'~ Netherlands 65/0S818 ~Farm~o~
1~306) discloses the react~Lon H2N~S
J=N~cl~2-s~cl)-ph ~ CH2COCl : :
COOH ~ ~
:, .
~
~ N \ GH2-S-CO-Ph AT1 ~quiv~ler~ U, S . 3 " 502, 665 .
' ~ ., : ~ /
~ ~ ~2 7~ ~
5. Gl~xo's Netherlands 64/11521 ~Chem. Abstr. 9 63: 13,281d (196S)] discloses the reac~ion ~ep~3 C ~in~5i~L~ HOOC-CH (CH2)3CONH ~ S ~
NH2 ~ ~ N ~ CH2-y COOM
whereln Y -S-CO-Ph or -S-CO ~
Equ~valent~ are Great Britain 1,101,422 and Can&da 796,747 (Farmdoc 17362).
6. Cib~'~ U.S. 3,555,017 disclo~es compounds having the structure . R
: R / C~-CO-NH ~ ~
COOH ~;
Rl ~nd R2 ~ halogen. ;- :
As usual in Ciba 18 patents directed prim~rily to novel ~id~ch~ins ~t the 3-posltion, the R group ~bove i~ defined broadly ~s the residue o~ ~
c~rboxylic acid and m~y be illustrated by phenyl, the residue of thi~benzolc acid. Equlv~lents ~re Belgium 7O~D24~ rmdoc 33~276), Great Britain 1,211,747 ~nd French 19575~554.
O g O
.... . ... . . .. .. . .. .. . . .. . .
~L~627(~
7. Ciba 's British 1,211~718 discloses com-pounds havlng the structure r O ~ C ~S .
R ~s in 6 above.~quivalents are Belgium 708,311 (Farmdoc 339277) and U.S. 39557,104. ::
8. Ciba 's Belgium 7519526 ~Farmdoc 90,178R) d~ sclo~es compounds having ~the s~ructure N3~ H2C-NH~ S ~
SC-R
COO~I ' ':
R a~: ~in~ 6 above.
An equ~valent i~ Netherland~ 70/08237.
:~ : 9. Ciba's South Africa 69l8436 di3closes : -. : compound~ having the structure `, ~N ~U ~C~ ~/ ~ O ". ~.
N ~ ~ S~
COOH
R a~ in 6 above.
.
E~ivalents ase Belgium 743,014 (Farmdoc 43,126RS
and ~etherlan~ds 69l18611.
.
~OÇ;Z7~ :
10. Ciba's ~outh Africa 69/8399 disc1ose8 compounds having the ~tructure N~; N~C \ CO-NH
~,==1 '~ H2 COOH
R a~ in 6 above.
Equlva1erlt~ are Bel~ium 742,933 ~rmdoc 41,568R) ~nd Netherlands 69/18531.
11. Giba's South Africa 68/8185 disc1oses compounds having the struc~ure H2 ' 0~ H2 R as ~n 6 above, COOH
An equivalent ~s Netherland~ 68118868.
12 . Ciba ' s Netherlands 68/18868 d~ sclose~
compounds having the structure _~ H2 : ' .
N ~ V~ O-NH
N~ ~ SC.-R
OOH
.
R a5 in 6 abov~. ~
Equivalents are South Africa 8120t68~ German 1,817,121 and Belg,lum 726~316.:
: ': -... ....... .. . . . . ..... .. . .. . .. .. : . . .. . .
~(~6~7(~1 13, Fu~isawal~ Great Brital~ 19187,323 for example, ~ page 5, lines 67-71 disclose~
compounds having the structure - ..
~ O
o\/ ~ Z ~ ~ S ~
R / R O / ~ / SC-R
, COO~
in which R represents ~ethyl, thienyl, pyridyl9 etc. ~nd wherein general disclosure is m~de of other heterocycllc groups a~: at pages 1 and 2.
Equivalents are Netherlands 67/14888 (Farmdoc 31,936) and U.S. 3,530~123.
.
~ 14. Fu~isawa~ Belgium 714,518 (Farmdoc 359307) discloses ~among many others) compounds having:the ~tructure R . :.
/ ~ B ~s 1l C-R
~ ~,CO~
w~ereln~ is as 13 above.
Equivalents are Netherlands:68/06129 and South~African 2695/68.
, ...
:: .
~ - 12 - ~
.. ... . . . , . _ . _ , . .. . _ . . ., . . _ . .
3L[36;27ai1 15 . Gl~xo ' s U . S . 3, 243 ,435 and Belgium 650,444 (Farmdoc 15,535) disclose generally a va~t variety of compounds h~ving the structure Acyl-NU-CO-llH~ CH~S
COOH
wherei.n ~, is defined, e.g. in column~ 1 and 4, t~ ~nclude various heterocycllc groups.
16. Cibals South Africa 65/6950 (Farmdoc 22 ~192) discloses compounds having ~he struc~cure N~ ~ C~lR2-~-N~S
,~N )~ SC-R
.` ~00~
in ~ich R in Example 20 is phenyl.
Equivalen~s ~re Great Br~tain 1,109,,525 ~nd Canada 807 ,651.
~ 17 . Glaxo ' s U. S ~ 3 ~47g , 350 d~sclo~os ~ proces~
for producing 3-pyridiniuImnethyl cephalosporin~
which utilizes ~s ~n intermediate compounds of the type~deser~bed in rcference~ 2 and 3 above~
' :
.
13 ~
~62710~L
18. Ciba 's U S. patent 3,7579013 d~scloses compounds of the formula ' 2~ ~\~ CH2-S-C-R
COO
wherein R3 i~ (lower3 alkyl and R is as in 6 above.
~ Lg. Bristol9s Belgi~n patent 795,811 disoloses compounds of the general fonn~lla Il ,S O
Ar- l~-C~ ~ rr NH2 `o~l N ~CH2-S-C-Z
COQR
wherein ~Ar is phenyl" 2-th~enyl or 3-thienyl; R
~ : : ls hydrogen, pivaloyloxymethyl, acetoxyme~chyl, ; methoxyme~hyl, acetonyl or phenacyl; and Z is ~.
H3 , _ N
~CU3 ~
~: 3-i~oth~zolyl 9 4-isothiazolyl or 5-isothiazolyl .
:
- 14 - ~
..
. . . ... .. ............. ., .. .. . .... . . . . . .. . . ....... ... , . . . .. ~ .
~9627~
This specification comprises the novel compounds having the D-configuration in the 7-sidechain and the formula ~r-CH-C-NH
~H2 ~ ~ ~ C~2~
COOR ::
wherein Ar is phenyl, 2-thienyl or 3-thienyl; R is hydrogen, :~ -pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl; and R' is 2-thiazolyl or 5-methyl-2-thiazolyl or -~
3-methyl-1,2,5-oxadiazol-4-yl, and their pharmaceutically acceptable salts. ;~
Summary of the Invention In a broad aspect the present invention provides a process for the preparation of a compound having the D- .
configuration in the 7-sidechain and having the formula `
H-I-N~ ~ S ~ 9 (I) H2 ~ N ~ C~ C-R' eOOR
: ~ .
whereLn R is hydrogen, pivaloyloxymethyl, a~etoxymethyl, : met oxymethylp acetonyl or phenacyl; and R' is 2-thiazolyl or 5-methyl-2-thiazolyl or 3-methyl-1,2,5-oxadiazol-4-yl; or a `
pharmaceutically acceptable salt thereof, comprising:
~a1 when the starting material includes the desired final substituent in the 7 position of the cephem nucleus:
reacting a compound of the formula `--Q ~ .
~ fH-C-NH 3 NH2 ~ ~ CH~OCOCH3 C2~ :
15 - ~
.. .. . . . ~ ~.. . . ~ .. . .
~(~36Z 17~L
or an easily cleavable ester or salt ~hereof wi.th a hetero-aromatic thiolcarboxylic acid of the formula o R'-C-SH
wherein R' is as defined above or a salt thereof to form the desired compound of formula I, or , ' (b) when the starting material includes the desired ~:~
final substituent in the 3-position of the cephem nucleus~
10 reacting a compound of the formula , ' ~2N ~ S ~
O N ~ CH~S-C-R' CO2~ .
:
wherein R' is as,defined above or an leasily cleavable ester or .~
salt thereof with acid having the formula '. ' .. ~... ...
CH-COOH
I
NHB ' '' or an acylating derivative thereof, wherein B is an Emino-protecting group and removing said amino-protecting group B to : :
produce the desired compound of formula I or an easily cleavable ester or,pharmaceutically acceptable salt thereof; -' . -':
and, if dPsired, in the case of (b) either before or after ~, removal of protecting group B ~i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or ,.
(ii) converting the product in the form of an easily cleavable ~ -'15(a) ~
. - . ... .
:
. .:
7~
ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
The pharmaceutically acceptable salts referred to above include the nontoxic carboxylic acid salts when R is hydrogen, e.g. nontoxic metallic salts such as sodium, potassium, calcium and aluminum, the ammonium salt and salts with nontoxic amines, e.g. trialkylamines, procaine, dibenzyl-amine, N-benzyl-pLphenethylamine, l-ephenamine, N,N'-dibenzyl-ethylenediaminel N-alkylpiperidine and other amines which have been used to form salts of penicillins. Also included within the definition of pharmaceutically acceptable salts are the nontoxic acid addition salts (amine salts) of the acids and ~
esters of Formula Ia, e.g. salts with mineral acids such as hydrochloric, hydrobromic, hydroiodic, phosphoric and sulfuric and salts wi~h organic acids such as maleic, acetic, citric, oxalic, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic and malic.
: . ...:
- 15(b) -.. ~, . .
1~627~
Preferred c~pounds of the present lnvention ~re the eompounds h~ving ~he D-configuratlon ln t:he 7~idech~in ~nd ~he formula Ax ~CH-C-NH
N~2 ~ c~2_s_~
COOR
Ib wherein Ar i6 phenyl, 2-thieriyl or 3-thienyl ~nd R iz hydrogen, p~v~loyloxymlethyl~ acetoxymethyl, ~ethoxyme~chyl s acetonyl or Iphenacyl; and the ph~rmaeeutically aeceptable salts thereof.
Preferred compounds of formula Ib are those in which ~9r i~ phenyl. An e~plecially preferred ~ompound of formula Ib ~ tElat in which Ar i8 ph~nyl ~nd R i9 hy~rogen; or the sodium or potas~ium s~lt ~her~of.
More preferred compounds of the present invention are the compourlds having the D-con-figuratiorl in the 7-sldQch~in and the fo~mula Q
Ar -CN-C -NU ~l CN2 _ 5 _C ~ CU3 COOR
. Ic wherein Ar i~ phenyl, 2-th~enyl or 3-thienyl and R i~ hydrogen, pivaloylox~nethyl9 acetoxy-methyl, methoxymethyl, ~cetonyl or phenacyl 9 ~nd the ph~rmaceutic~lly ~ccept~ble salts thereo~. Preferred compound~ of formul~ Ic are tho e in whieh Ar is phenyl. An especi~lly ~L~627~
preferred compound of formula Ic is that in which Ar is phenyl and R is hydrogen; or the sodium or potassium salt thereof.
Further preferred compounds of the invention are the compounds having the D-configuration in the 7-sidechain and the formula A~ N~ H -5-~
Id wherein Ar and R have the meanings given above and the pharmaceutically acceptable salts thereof.
Preferably Ar is phenyl and R is hydrogen; or the sodium or potassium salt thereof.
Also included in this invantion ~re the co~pounds ~u~ed as sither int~ermediates or met~-bolie precursors) in ~hlch thle amino group i8 "blocked" by substituents suclh a~ t-butoxycarbonyl, c~rbobbn~yloxys~formy~ o-nitrophenylsul~enyl9 -trichloroethoxycarbcnyl~ 4-oxo-2-pentenyl-2, l-c~rb~me~hoxy-l-prspenyl-2 snd the like. P~rtic-ularly lncluded in ~uch blocking groups are the ketones (especially acetone) snd the ~ldehydea (espec~ally formaldebyde ~nd acetaldehyde) di~-closed for example ln U.S. patents 3,1989804 ~nd ~347,851 and the ~ketoesters ~nd the.~-diketones di~closed for ~x~mple ln U.S. patent ~9325,479 ~nd the ~ketoamides disclosed in Jap~n 71/24714.
The pr~ent in~ention also includes a process for the p~eparation of ~ eompound having the formula p A~_q~ ~T~S
H2 ~ ~ CH2-S-C-R' CC)~
' wherein Ar is pheny~ t~ienyl or 3-thienyl;
R' is 2-thiazolyl or 5-methyl-2-thiazolyl or 3-methyl-1,2,5-oxadiazol-4-yl; and easilY cleavable esters and pharmaceutically acceptable salts thereof;
which process comprises either ~A) reacting ~ compound o~ the formula ., M2N --- I , s ~
o~ ~CH2S-C~R' II
_._ wherein R' is as defined above or an easily cleavable es~er or salt thereof with an acyla lng deriv~tive of an acid having the for~ula :
Ar-~H-COOH
~HB .
III `
, ~ , .. .
wherein B is an amino~protecting group and Ar ~s a8 defined above and removing sa~d amino-protecting group B to produce the desired com-pound o formula I or an easily cleavable es~er or ph~rmaceut~cally acceptable s~lt thereof and, if deslred~ ei~her before or after removal o~
pro~ecting group B (a) converting by methods -known ~ se the product ~n the form of the free acid or 8alt the~eof to a corresponding e~sily cleavable es~er or pharm~ceutically ~ccept-able sàlS thereof or ~b) converting by methods known~per se the product in the ~orm of an easily cleavab1e ~ster or ~alt thereof to the correspondlng free aci~d compound or pharmaceutically acceptable s~lt thereo~; or , .
~8 -.
l~Z7~1 (B) reac~ing a compound of the formula Ar-CH-C-NH ~ ~
~ 2 0 ~ ` ~ CH2ococH3 IV
whereln Ar is 3S defined above or an easily cleavable es~er or salt thereof with a hetero- `
aromatic thiolcarboxylic acid. of the formula R'-C-SH
:~ .
where~n R' is as defined abo~e or a s~lt thereof to form a compound of formula I or an easily cleavable ester or pharmaceutically acceptable 8alt thereof and, if desired, (a) converting by methods known ~ se the product in the form of the free acid or salt thereof to ~ cor~esponding easil~ cleavable ester or pharmaceutieally accept- :
able salt thereo~ o~ (b~ converting by methods `
~nown ~ se~the product in the form of a~n easily cleavab-le ester or salt thereof to the corresponding free acid compound or pharmaceutically ~cceptable sal~c ~ thereof.
. .
The easily cleavable esters referred eo above include ester groups which are removable by methods, e.g. chemical or:enzymatic hydrolysis~ which do not result ~n any appreciable destruct~on of the remaining~-portion of the cephalosporin molecule.
~xamples of suitable esters include those disclo~ed in U.S. patents 3,284,451 ~nd 3,249,622 and U.K.
patents 1,229,453 and 1,073,530. Parti~ularly . .
- 19 - ' , ~ ~ ~2 7~ ~
preferred esters are the piv~loyloxymethyl, acetoxymethyl9 methoxymethyl, acetonyl and phen~cyl esters.
In one method of preparing the novel cephalo-~porin compounds of the present invent~on, a 3-thio-lated-7-aminocephalosporanlc acid compound of form~l~ II or an easily cleavable ester or sa1t of ~id acid or ester 18 acylated w~th ~he appro-priate acylating derivative of formula III.
The 3^thiolated-7~amlnocephalospor~nic ~cid lntermed~a~e of formul~ II may be prepared by - di~placement of the 3-ace~oxy group of 7-amino-cephalosporanic acld or a salt ~hereof wi~h the appropriate heteroaromatic thiolcarboxylic ~cid or g salt thereof. The displacement of an ester group with a thiol group is a known reaction and i~ preferably accomplished iLn aqueous solution at a temper~ure oi ~t. least room temperature and preferably within the range of about 50 to 100C.
. iLn the presence of ~ mild base such as sodium b~carbon~te.
Th~ claimed compounds may then be vbta~ned ; by a~ylatlo~ ~ccording ~o known methods of the 7-amino group o~ intermediate II with the acyl3ting agent of`formula I~I.
: Because of the low solub~lity of the compound~
of formNla II in common aqueous an~ non-aqueous : 801~ent~, lntermediate II ls preferably converted prior to:the acylation reac~ion ~o an`easily cte3va~1e~ester or ~cid~addi~ion salt thereofo The procedures for preparing such es~ers are d~sclo~ed in the literature ~nd are well-known t~ those skilled in the art of penicillin and cephalosporin chemistry. One preferred me~hod espec~ally useful for preparing 'ch~ most preferred e~sily hydrolyzed esters, i.e. the piv~oylox.ymethyl, - ~0 - ;
, . .. .. . . _ . . _ .. . , _. . . . . . ..
1~6Z~7~1 acetoxymethyl 3 methoxymethyl, acetonyl and phen-acyl esters, is disclosed in U.S. patent 3,284,451 This reference descrlbes the esterification of sodium.cephalothin with the appropriate active chloro or bromo compound (e.g. phenacyl bromide, ~hloroacetone, chloromethyl methyl ether, pivaloyloxy;
methyl chloride, acetoxymethyl chloride3 followed by enzymat~c removal o~ the thienylacetic acid ~ide-ch~in. In another good method ~he ~rie~hylamine salt of 7-aminocephalosporanic ~c~d i~ reacted directly with the active halogen compound as ln U.K. patent 1,229,453. The compound of formNls II n~y also be converted to ~ ~ilyl ester as by the methods described ln the literature, e~g. .
U.S. patent 3,249,622. The silyl ester group may be removed following the acylation reaction : .
by hydrolysis or alcoholysls. ~ -Prior to the ~cyla~ion reaction the amino group o~ acylating agent III is protected by a conveneion~l amino-protect-img group B which may be read~:ly re~oved ~t the conolusion of the react~on. Examples of suitable amino-protect~ng groups include tbutoxycarbonyl, carbobenzyloxy, 2-hydroxy-l naphthcarbonyl, trichlor~ethoxyc~rbonyl, 2-ethoxycarbonyl-1-methylvinyl and 2-methoxycarbonyl-l-methylvinyl. A p~rticularly valuable ~locking group~is;a prOtOII, as in the compound of the formNls .
Ar-~H-COCl wherein Ar i~ phenyl, 2-thienyl or 3 thienylO The preferred amino-protecting groups are t-butoxy-carbonyl, the proton and a ~-diketone or a ~-keto~ -ester as in U.K. paten~ 1,123p333 or U.S. patents .
.. ... . .. ... . . .. . , . . . . _ .. . . . . . .
~L062~
3,325,479 and 3,316,247, e.g. methyl acetoacetate9 or a ~^ketoamide as in Japan 71/24714. When the t-butoxycarbonyl~ ~-ketoester, ~ diketone or ~-ketoamide protecting groups are employed~ it ls preferred to convert the acylating acid con-taining the blocked amino group to a mixed nnhydride~ e.g. with ethyl or isobutyl chloro-formate, before reaction with compound II or an e~ter or ~alt thereof. After the acylàtion coupling reaction, the amino-protecting group B
m~y be r~moved by methods known ~ se to form the desired product of formula I. Thus, for example, the t-butoxycarbonyl group m~y be removed by use of formic acid, the carbobenzyloxy group by catalytic hydrogenation, the 2-hydroxy-1-naphth-carbonyl~group by acid hydrolysis, the trichloro-ethoxycarbonyl group by tre~stment with zinc dust ln glacial acetic acid, ~he proton by neutrali-zation, etc. Obviously other func~ion~lly equ~v~
alen~ blocking groups for an amino group can be used and such groups are considered within the scope of this invention.
Acylation of a 7-~mino group of a cephal-~sporin ~ 8 a well-known reaction and ~ny o~ the func~ional equivalents of formula III commonly used ~s acylating agents ~or primary amino group~
m~y be employed. Example~ o sui~able acylating derivatives~of the free acld include the co~re5-ponding ~cid anhydride~, mixed anhydr~des, e~g.
alkoxyformic anhydrides, ~cid halides, acid az~des~
~ctive esters an~ active ~hioesters~ The free acid m~y be coupled wi~h compound II ~fter first reacting said free acid with N,N1-dimet~ylchloro-~ormimin~um chlor~de or by the use of enzyme6 ~r .
'~ ' ''' .
, . . .. . ... . . . . .. . ... . . . . . . .. .. . .. . .. . . . .
~ 0 6Z 7 of ~n N,N'~carbony1diimidazole or an N,N' c~rbonyl~
ditr~azo1e or a c~rbodiimide reagent, e.g. N,N!-diisopropylcarbodiimide. N9N'-dicyclohexylc~rbodi-imide or N-cyclohexyl-NI ~2-morpholinoethyl)~
c~rbodiimide or of alky1ylamine reagent or of an i~oxa~olium salt re~gent. Another equivale~ of ~he free ac~d i~ ~ corre~ponding azollde, i.e., an amide of the corresponding acid whose a~ide nltrogen i~ a member of ~n quasiaromatic five membered ring containing ~t least two nitrogen ~toms, ~.eO 3 i~idazole, pyrazole, ~he triazoles, benzimidazole, benzotri~zole and their ~ub~tituted deriv~t$ves. Another react~ve derivat~ve of the pheny:lglycine acid of formNla III i8 thè N~carboxy anhydride (Leuch'~ anhydrid~) . In this structure ~he group which activfltes the earbo~cyl group also serves to protect the ~m~no group. A particularly preferred acylating agent i8 the acid chloride hydrochloride of the iEormula Ar-~H-eOCl NH2 ~ HCl ,.
whioh also ~erves ~ du~l function of osrboxyl ~ctivation and amino proteotion. Mention was m~de above of the use of enzymes to couple the free ~cid with its blocked amino group wi~h c~m-pouFad II. Included in the scope of 6ueh proce~ses ~re ~he use of an ester, e.g. the methyl ester, of that free acid with enzyme~ provided by v~riolls microorg~nisms, e.g. those described by T. Takahashi et al., J~ Amer. Chem. Soc., 94(11~ 9 4035-4037 (1972 ~nd by T. Nara e~ ~1., J. A~tibiotics (Japan), 24(5), 321-323 tl471) and ln West Germ~ny 292169113.
:: - , . .. , .. ,. . , .. , . ~ , .
~ 0 6~ ~0 ~
The particular proce~s conditions, e.g.
tempersture~ solvent, react~on time, e~c. ~elec~ed for the acylatlon coupling reactlon are determined by the nature of the acylation method used and ~re known to those skilled in the art. Generally it i~ useful to add an organic tertiary amine, e.g. tr~ethyLamine, N~N-dimethylsniline, ethyl-piperidine, 2,6-lutidine or quinoline, to ~erve as a proton acceptor or salt-forming agent. A
preferred method ~llustrated in the examples which follow involves formation of a s`ilyl ester, e.g. with trimethylchlorosilane, of the inter-mediate of formula II and acylation of this sily~
l~ted intermediate ~n dry methylene chloride at temperature of below room temperature ~nd prefer-ably about 0C. with the appropriate chloride hydrochloride acylating age!nt of formula III in the presence of ~n organic tertiary am~ne.
At ~he concluæion of the acylation reaction, the acylated intermediate is sub~ected to aqueous hydrolys~s to provide the desired cephalo~porin produo t .
~ The eompounds of the present invention m~g be isolated in any of the ways ~ustomarily empIoyed for ~he isolation of similar cephalos-porin~. Thus 9 the product may be ob~ained ~ -~he neutral moleeule~ although thi~ is prvbably more~accurately represented ~s the æwitterion, or ie m~y be ~solated as a ~al~. Form~tion of the de~ired pharmaceutically aceepta~le carboxylic acid or acid addition salt ~s carried out by known methods, e.g. reaction of the ~cid with ~n appro~
pria~ base or acld. ~ ~ :
At the conclusion of the acylation reaction ~he product obt~ned m~y be converted (before or 1 0 6 2 7~ ~
after r~moval of the ~mino-proteceing group) by methods known per _ to another desired product of formula I. Thus, the compound of formwla I
in the orm of the free acid or a s~l~ thereof m~y be converted by known methods ~o a corres~
pondlng easily cleavable ester or pharm~ceutic~lly ~ccep~able s~lt thereof. Similarly, the product of formula I in the form of ~n eaælly cleavable ester or s~f~ thereof may be converted to the ree acid product or pharm~ceutically ~cceptable ~alt thereof by removal of the esterifyi~g group9 e.g~ by ~queous or enzymatic hydrolysis ~a~ wlth human or ~nimal serum~ or acidic or alk~line hydrolysls or by catalytic hydrogenation or by treatment with sodlum thiophenoxide as t~ught in U.S. patent 3,284,451.
In another method of preparing the compounds of the present invention, 7-am~nocephalosporanic ~cid or a salt thereof is acylated with the acid oiE formul~ III or an acylating deriv~tive thereof to fo~m ~ 7-acylated cephalosporin compound of the formula ~ .
Ar-lH_ 3 ~H - -- f ~
NH2 ~ ~ CH20COCH3 CO2~' , ., . I~
Compound IV in the form of the free acid or an easily cleav~ble ester or salt thereof is then reacted ~ccording to the process of the present invention with ~ heterocyclic thiolc~rboxylic acid of formul~
V or a salt thereo~ mos~ preferably the sodium ~L~6Z7~1 or potassium salt. The displacement resction i8 preferably conducted in aqueous solution ~t temperatures of about 50CO or higher in an inert atmosphere, e.g. under nitrogen. The product of the displacement reaction m~y, if desired~ be converted to a pharmaceutically acceptable salt by treatment with an appropr~ate acld or base. As in the ca~e of the alternate process described above for prepara~ion of the compounds of formNla I, the product in the form of the free acid or salt thereof may be converted to a corresponding easily cleavable ester or pharnaceutically acceptable salt thereof or, alternatively, ~he product in the form of an easi1y cleavable ester or salt thereof may be .
converted to the free acid or pharmaceu~ically scceptable salt thereof.
The easily cleavable esters of the compound of formula I are use~ul as i.ntermediate8 in the produc~lon of the free ~cid product. The piv~l-oyloxymethyl 9 acetoxymethyl and methoxymethyl esters are al~o useful as active antibac~erial agent~s 6ince on oral administration they are rapldly~hydrolyzed ~o the active me~aboli~e.
These three esters are of particular ~nterest because~ they provide on oral administration different ra~es and amounts of ab~orption and give differing concentrations of ~he active anti-bacterial agent in blood and tissues.
The preferred and most active compounds of ' the presen~ invention ~re those having the D-configuration at the a-carbon atom in the 7-side- l chain, that is, those m~de from D~ 2-phenyl-~
glycine, which is also called D-~ a-amin~phenyl- .
acet~c ac~d,.and D-(-)-2-thienylglycine and D-(-~-~062~C)1 3-thienylglycine. In addition, the conf~gura-tion at the two optically active asymmetric centers in the ~-lactam nucleus is that found in cephalospor~n C produced by fermentation and in ~he 7-aminocephalosporanic acid derived there-from.
The pharm~ceutically active compounds of the present invention are potent antibacterial :
~gents useful in the treatment of infertious dlseases in poultry ~nd animals, including man, caused by m~ny Gram-posi~ive and Gram-negative bacteria. The active compounds are also of value ~s nutritional suppllements in animal feeds .
and as agen~s ~or the trealtment of mastitis in ca~le. The preferred compounds have also been unexpectedly found to be efficiently absorbed uposl oral administration.
Despite the synthesis of a large number of cephalosporins reported in the scientific and patent liter~ture, the only cephalosporin at th~
present ~me having suffic;Lent actlv~ty ~nd or~l ~bsorption for general: oral use is cephalexin.
In our search for new orally active cephfllosporins, we ha~ve found ~hat the compounds claimed~ in the present ln~ention fiurprlsingly and unexpectedly possess:~good or~l absorption and at ~he~lsame ~ime appear to be superior to cephalexin ~n the~r activity against certain impor~ant pathogen~c org~ni~ms. Thi combinat~on of good oral absorp-tion~and high antibacterlal ace~vity~ln a~ceph-~losporin while highly desirable is quite uncommon in compounds reported to date. M~king~any reliable predict$on as to artivlties or oral ~bsorptions of new cephalosporin compounds from a~ examLnation of .
~62~
the propert~es of structurally analogous prlor ~rt compound~ has also b~en found to be essen-tially impossible. To illustrate the unpredic-t~bill~y of oral absorption and antibacterial activity in ~ closely related series o~ cephal-o~po~in compounds 9 Tables 1-8 below compare in vitro activitie~ ~nd oral absorption as tes~ed in the mouse of the compoun(l~ of the formula H_~_N~ S
NH2 ~ CH2-S-C-Z
D~
wherein Compound No.
.. . .
æ ~ 2-thlazolyl 567
- 4-th~azolyl 591
5-~hiazolyl ~02 4-me~hyl-2-~h~azolyl 556 5-methyl-2-~hiazolyl 586 2 methyl-4-thiazolyl 607 - 4,5~dimethyl-2-thiazolyl 587 , Tab~es 1-8 inslude data on the commercial or~l cephalosporin, cephalexin.
' .
.... . . .. , .. ... ....... . . ..... . .. . .. " . . . . .. . .. .
-. :
1~627~1 Samples of the seven cephalosporin compoundc having the compound numbers reported above after ~ol~tion ln DMSO ~dimethyl sulfoxide) at 14 mgm./
ml. followed by dilution with Nutrient Broth were found to exhibit the following Minimum Inh~bitory Concentrations ~M.I.C.) ~n mcg./ml.
versus the indicated m~c~oorgan~sm~ as determined by overnight incubation ~t 37C. by Tube Dilution.
The in vitro activity of cephalexin W9S also tested.
~.
- : -',~`. ' ~: . ...
~ , : .
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.
~L0627~:~
T~ble I
~i. 1. C . in mc~ . /ml .
Compound Cepha-r~anism No. 586 lexin D .pneumonia e ~ A9585 .16 . 6 5% serum *
Str . pyogenes + A9604 . 3 . 3 5/O serum *
S . aureus Smith $ A9537 . . 3 1. 3 S . aureus Smith + A9537 >63 2 . 5 50% s erurn ~: :
S . aureus PsX 1633-2 A9606 . 6 4 ~ t 10- 3 d ilution -S. allreus BX 1633-2 A96n6 4 8 at 10-2 dilution S. aureus methicil~in- A15t)97 4 32 resis~ant ~t 10- -dilution .
S. aureus at 10-3 A9748 8 32 dilution S. aureus~ at 10~ 2 A9748 63 l25 dilution Sal. enterltidis $ A9531 ,3 4 E. coli Juhl ~: A15119 2 8 E. coli ~ A9675 4 16 ~, pneum~ni~e $ A9977 4 8 K. pneumoniae ~ A15130 3~ :~ 16 ~ .
Pr . mirab~lis =~ A9900 16 ~ ~ 4 Pr. morganii ~ A15153:63. ~ 125 Ps . aeruginosa ~ A9843A>125 ~125 Ser. marcescens ~ A20019>125 ~ >125 .
* 50% Nutrient Broth _ 45D/o An~ibiotlc Ass~y Broth :~ ~t 10-~ dilu~ion ~ 30~
~L~6270~3L
Table II
M. I . C . in m~ . /ml .
Compound Cepha-Organism No. 567 lexin D . pneumoniae ~ A9585 . 04 5% s erum*
Str . pyogenes ~ A9604 . 04 . 3 5% ~erum *
S, aureu~ Smith $ A9537 ~3 o6 S. aureu~ Smith ~ A9537 8 2 5 50h ~erum 7 -S . aurèu~ BX 1633-2 A9606 . 6 2 ~t 10-~ dilution S. aureus BX 1633-2 A9606 . 2 4 at 10-2 dilution S. aureus methicillin- AlS097 4 32 res is tant a t 10-3 dilution S. aureus at 10-3 A9748 4 32 dilution S~ aureus :at 10-2 A9748 16 125 dilution Sal. enteritldis ~: A953i - .08 2 E. coli Juhl ~ A15119 1 ~ 8 E. coli ~:: A9675 b,~ lS
K. pneumoniae~ A9977 2 4 K. pneumoniae* A15130 8 16 ~' ,' . .
Pr. mirabilis ~: A9900 2 ~ 4 Pr. morganii $ A15153 63 ~ ~ ~125 Ps. aeruginosa ~ A9843A >125 ~125 Ser. marcescens ~: A20019 125 >125 * 50%~Nutrient Broth ~ 45% Antibiotic :Assay Broth :~ at 10-4 dilutLon Table III
M. I . C . in mcg . /ml .
CompoundCepha-~ No._591lexin D . pneumoniae ~ A95B5 . ~2 . 3 5VL serum *
Str . pyogenes ~ A9604 . 02 . 6 5% serum *
5. aureu~ Smith ~t A9537 .3 . 1.3 S. aureus Smil:h + A4537 8 2.5 50% serum *
. . .
S. aureus BX 1633-2 A9606 .6 4 ~t 10-3 dilution S. aureus BX 1633-2 A9606 2 4 at 10-Z dilution S. aureus methicillin- Al5lD97 2 63 resistant at 10-3 dilution S. aur~us:a~ 10-3 A9748 4 63 dilution S. aureus: ~t 10-~ A9748 16 125 dilution ~
Sal. enteritidis $ A9531 .16 4 1~. coli Juhl $ A15119 1 ~ ~ 8 E, coli ~ A9675 4 ~ 32 K. pneumoniae ~ A9977 ~ 1 4 K~ pneumoniae $ A15130 8~ ~ 32 Pr. mirabilis ~ A9900 ~ 4 Pr. morgani~ * A1515332 ~125 Ps. aeruginosa ~ A9843A>125 : ~125 Ser. marcescens :~ A20019125 ~ >125 * 50% Nutrient Broth ~ 45% Antib~otic As~ay Broth * a~ 10-4 dilution -.. . . . .. .. . ... . . . . .. . .... . . . . . . ... .
~L06Z7(;1~
Table IV
M I.C. in mcg./ml.
Compound Cepha- I
Or~nlsm No. 607 lexin D. pneumoniae ~ A9585 .16 .16 5% serum *
Str. pyogenes + A9604 .16 .16 5% ~erum *
S. aureus Smith ~ A9537 .3 o6 S. sureus Smlth + A9537 63 1.3 50% serum ~ I
S. aureus BX 1633-2 A9606 .6 2 1 .
~t 10-3 dilution S. aureua BX 1633-2 A9606 4 8 ~l ~t 10 ~ dilution S. aureus methlcil3in~ A15097 8 63 .
resistant a~ 10-dilution S. ~ureus at 10-3 A9748 8 63 .
dil~tion s. Aureus At 10 ? A9748 16 125 ~
dilution ~ . .
Sal. enter~tidis $ A9531 .6 4 .
: ~ E. coli Juhl $ A15119 8 . 8 .
E. coli . ~ ~ A9~75 16 ~ ~ 16 ~
K~ pneumoniae ~ A997716 .~ 4 .
: ~ pneumonia~.i A15130125 ~ ~ 16 .
Pr. mirabills $ A9900 `8 ~ ~ 8 Pr. morgan~ $ A151533~ : >125 p~. aerugiDosa $ A9843A~125 - >125 -Ser. marceScena ~ A20019~2$ ~125 * 50% Nutrient Bro~h - 45~ ~ntibiotic Assay Broth .
$~t 10-4 dilu~on - ~
~ ' . ' ,. ~ 33 ~1~627~
Table V
M.I.C. ~n mc~. /ml.
Compound Cephs-Or~anLsm No. 602_ lexin_ û ~ pneumon~ ae ~ A9585 . 60 3 5% serum *
S tr . pyogenes -~ A9604 ~ $ . 6 S% serum *
S. ~ureus Smith ~ A9537 ,3 1.3 S . aureus Smith + A9537 >63 2 . 5 50% ser~m ~
S . aureus BX 1633-2 A96061. 3 4 st 10-3 dilu~ion S. aureus BX 1633-2 A960616 at 10-2 dilution . S. aureus methicillin- A15097 4 32 resi~tant at 10-3 d llutlon S~ au~eus at 10-3A9748 8 32 ~ilut~on S. aureus at 10-2A9748 63 . 63 dilutiorl Sal. enteritidis ~ A9531 . 3 4 E. coli Juhl ~: A15119 ~ :~ 8 E. coli ~ ~ A9675 16 32 K. pneumonlàe ~ ~ A9977 16 8 K. pneumoniae ~ A1513032 ~ 16 Pr. mirabllis i A9900 16 8 Pr. morganii =~ A15153125. ~125 P8. aeruginos~ ~ A9843A>125 >125 .
Ser. marcescens $ A20019 ~12$ ~125 ~k 50% Nutrient Broth ~ 45% Antib~ot~c Ass~y Broth :
~ ~t 10-4 dilution ~ ¦
~ 34 -~a~6~7 Table VI
M . I . C . in mc ~ . /ml .
C:ompoundCepha-m No. 587lexin D O pneumon~ ae ~ Ag.~85 2 . 5 . 3 5% serum *
Str~ pyogenes + A9604 2 . 5 1. 3 S% serum *
S . aureu.s Sm~th ~ A9537 . 3 1. 3 S, aureus Smith ~ A9537 >63 2. 5 50% ~e~
S . aureus BX 1633-2 A9606 . 6 . 4 flt 10-3 dilut~on S. ~ureus BX 1633-2 A9606 1 4 ~t 10-2 dllutlon S. a~lreus methicillirl- Al5097 4 63 resistant a 10 3 dilut~on S. aur.eus at 10-3 A9748 4 63 dilution S. aureus at 10-2 A9748 1~: 125 dilution Sal. enterit~dis $ A9531 2 . 5 4 E. coli Juhl ~ A15119 I~ 8 E. coli ~ A9675 32 16 -K. pneumoniae =~ A997732 4 K. pneumoniae =~ A15130 ,~125 16 Pr . m~rabilis ~ A9900 >12 5~ ~ 4 Pr. morganii ~3: A15153 >125 >125 Ps. aeruginosa ~: A9843A ~125 . >125 Ser. marGes~cens ~ A20019 ,~125 ~ ~125 * 50/O Nutrient Broth - 45% Antibiotic As8ay Broth ~t 10-4 dilution :~CD62763:~L
Table VII
M~IoC~ in mcg./mlO
Compound Cepha-~ No. 556 lexin D. pneumoniae + A9585 1.3 .16 5% serum *
Str. pyogenes + A9604 2.5 .08 5% ~erum *
S. aureus Smith ~ A9537 1.3 .6 S. aureus Smith + A9537 63 1.3 50% s~rum ~ .
S. aureus BX 1633-2 A9606 1.3 2 ~t 10-3 dilution S. aureus BX 1633-2 A9606 4 4 at 10-2 dilution S. aureus methic~llin- A15097 4 32 resistant at 10-3 dilutlon S. aureùs ~t 10-3 A9748 ~ 32 : dilution s. sureus a~ 10-2 A9748 8 : 63 : : ~ dilution Sal. en~eritidis ~ A9531 ~ 2 E. coli 3uhl ~ ~ A15119 32 ~ 8 : E. coI~ $ A9~75 32 : 16 ~ .
K~ pneumoni~e ~ ~A9977 3~ 4 K. pneumoniae $: A15130 125 16 PL~. mirabllis =~ A99ûO 32 4 Pr . morganii ~ A15153 12 5 >125 ~s. aerug~nosa $ A9843A 125 >125 Ser. marceseens-~ A20019 63 >125 * 50~/~ Nutrient Broth - 45% Ant:~blot~c Assay Broth at 10-4 dllution :
1~6Z'7~
T~ble VIII
Blood levels in the mouse after oral admini~tration were determined with the following results:
. ~ , . .
- ._ Bloc d Level _in mc~. /ml.
fi~ S Dose O.5 1 2 3.5 ~CH-GONH--~ ~Hrg. aft2r ~dminis-I . l Imgm. /kg. tration.
~2 6 ~:H2R
.. . , ' ,. ~
-S-C ~H3 1.00 34 . 8 30 . 7 13 . 0 7 . O
., _ . .... ,._ , ., ' 11~3 100 l16.0:14.1 3.4<0.4 567 ~ : -. ~, ~ . ..
-s-~ ~
I ~ lo~ 6 s 7.9 4.4 3.2 591 : ~
_ .. . .... _~ - --~1CH3 ~ 1.2 1.4 1.3 1.0 , ~62~
Blood Level in ~g./ 1.
Dose O.S 1 2 3.5 Hrs. after adminis-m~m. /k~. tration _ ' ., . . ...
-sd~ oo Ill.7 17.6 9.7 3.3 602 . .
-S-d;~CH3 100 6.3 S.9 4.3 2.0 __ . . '':,~
I CH3 :.
-æ-~ ~sl 100 15.2 16.2 7.8<2.1 556 .
: _ . .
, . .~ .
-H 10044 . 6 ~ 3 . 4 6 . 1 1. 7 (cephalexin) ~onohydrate . -~:
~, ', ' ': : '' -.
.~ 8 ..
.... . , ... ... . . , . . . .... . . . . . _.. . ~ . .. ... .. . . . . .. . .
~627~
An examinat~on of Table VIII shows that very small struc~ural changes in the 3-acyl-thiomethyl substituent can greatly affect oral ~bsorption, The very closely-related compounds S86, 6~7 and 556,` for example, have blood levels after 3b minutes of 34.8, 1.2 and 15.2 mcg./ml.
An examination o~ Tables I - VII shows the same unpredictability with respect to in vitro activi-ties. Compounds 586 and 567 claimed in the present invention surprisingly and unexpectedly showed both excellent activity relative to cephalexin in the primary screening tests and good or-l absorption proper~ies.
.
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.
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i270 The novel medicaments provided by the present inven~on may be formulated as pharmaceutical com-posltions comprising, in addition to the active ingredient, a pharmaceu~ically acceptable carrier or diluent. The compounds may be administered both orally and parenterally. The pharmaceutical preparations may be in solid form such as capsules, ~ablets or dragees, or in liquid form such as solutlons, suspensions or emuls~ons. In the treatment of bacterial infections in man, the compounds of th~s invention may be administered in an amount of from about 5 to 200 mg./kg./day in divided dosage, e.g. 3 to 4 times a day.
They are administered in dosage units containing e.g. 125, 250 or 500 mg. of active ingredient with suitable physiologically ~ccep~able carriers or excipients.
In the experimental s'ections which follow, all temperatures unless indicated are in degrees Centigrade. The abbreviation 'tMeC12" is used to refer to methylene chloride while '~EA" is used to indicate triethylamine.
~' :
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' 1~627~
PreE~ration Of Starting Materials Thiazole-2-thiocarboxylic acid dicyclohexylamine salt A mixture of 14.2 g. (0.11 moles) of 2-carboxy-thiazole, 220 ml. of methylene chloride, 13.9 g.
(0.11 moles) of oxalyl chloride and 0.5 ml. of N,N-dimethylformamide was stirred for 3 hours.
The insoluble material was removed by filtration, washed with methylene chlorid~ and the filtrate was evaporated under reduced pressure.to a solid.
This obtained acid chloride was-added portionwise to a st~rred and cooled solution of 24.2 g. (0,22 moles) o sodium sulfhydrate trihydrate in 247.5 ml.:of ethanol and 27.5 ml. of water ~t such a rate as to maintain the temperature of the reaction mixture at 10-15. After the addition was completed, the mixture was stirred for 40 minutes at 5-10.
Then the ethanol was removed under reduced pres~ure and the residue was dissolved in 145 ml. of water.
The pH o the solution was lowered to 2.8 by the addition of 6 N hydrochloric acid, and n~intained there:while the mixture was ex~racted w~*h 5 x 200 ml. of ethyl acetate. The eombined extracts were.washed with ice water, dried over:MgS04, filtered and evaporated under reduced pressure $o a reddish solid. This was weighe~d,-redissolved ;~.
in 100 ml. of warm ethyl acetate and l:equivalent :-.
of dicyclohexylsmine was added. Bright yellow crystals start2d to precipitate immediately.
The mixture was cooled in an ice-bath ~or 1 hour.
Then the crystals were removed by filtra~tion?
w~hed with e~her and dried under reduced.pressure .:
, - 41 -. .
~627~
for 15 hours; weight 15.2 g. (42.5% yield).
Analysis:
Calcd. for C16H26N20S2: C, 58.88; H, 8.04;
N, 8.62; S9 19.60.
Found: C~ 58.47; H, 8.11;
N~ 8.74; S, 19.91.
7-Amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem 4-carboxylic acid Fifteen g. of thiazole-2-thiocarboxylic acid dicyclohexylamine salt was ~lurried in 175 ml. of wa~er and layered with 175 ml. of ethyl acetate.
The pH of the cooled solution was lowered to 2.2 by the addition of 6 N hydrochloric acid. The layers were separated and the aqueous phase was extracked with 2 x 175 ml. of ethyl acetate. The combined extrac~s were driecl over MgS04, filtered and concentrated to a red solid, weight 5.35 g.
The infrared spectrum indicated thioacid.
To a ~tirred solution of 9.8 g. (0.036 moles) o 7-aminocephalosporanic acid and 6.05 g. (0.072 moles) o~ sodium bicarbonate in 170 ml. of aqueous phosphate buffer at pH 6.4 was added 5.35 g. (0.036 moles) of thia201e-2-thiocarboxylic acid. The mixture was stirred in a nitrogen a~mosphere at 50 for S hours keeping the pH at 6.6 with 42%
phosphoric acid. Then the mixture was cooled to 20 and t~e precipitated solid was ~emoved by filtration, washed with water, acetone and ether ~-~nd dried in _cuo over phosphorus pentoxide to provide 4,6 g. (35% yield) of a tan crystalline solid. Infrared ~nd NMR spectra were consistent with the structure.
, .
~ ~ ~Z 7~ ~
Pivaloyloxymethyl 7-amino-3-~2-thiazol~l)carbonyl-thiome~yl-3-cephem-4-carboxylate_ Method A. The title compound is produced by substituting for the 7-aminocephalosporanic acid used immediately above an equimolar weight of pivaloyloxymethyl 7-aminocephalosporanate hydrochloride prepared according to Example 2 of U.K. 1,229,453 from 7-aminocephalosporanic ~cid. German 1,904,585 (Farmdoc 39,445~ ls equLvalent to U.K. 1,3399453.
Method B. The title compound is produced by substituting for the 0.025 mole (6.8 g.) 7-aminocephalosporanic acid used in the pro-cedure of Example 2 o~ U.K. 1,229,453 an equi^
molar weight of 7-amino-3-t2-thiazolyl)carbonyl-thiomethyl-~-cephem-4-carboxylic acid.
The respective acetoxymethyl, methoxy-methyl, acetonyl and phenacyl esters o~ 7-amino-3-(2-th~azolyl)carbonylthiomethyl-3-cephem-4-carboxyl~c acid are prepared by substitut~ng in Method B above for the chloromethyl pivalate used therein an equimolar weight of chloromethyl acetate, chloromethyl methyl ether, chloroacetone and phenacyl bromide, respectively.
5-Methylthiazole-2-thiocarboxylic acid dicvclohexvlamine salt A mixture of 14.3 g. (O o l mole) of 5-methyl-thiazole-2-carboxylic ~cid in 200 ml.~of dry methylene` chloride, 12.6 g. (0.1 mole) of oxalyl chloride~and 0.5 ml. of N,~-dimethylformamide . .
was stirred for 3 hours at room temperature until a clear solution resulted. It was concentrated to a greenish oil, which was identified by in-frared as the acid chloride. This was added ~ ~3 -, Z7~
slowly over 5 minutes to a s~irred and cooled ~olution of 22.0 g. ~0~2 moles) of sodium sulf-hydrate trihydrate in 225 ml. of 100% ethanol and 25 ml. of water at such a rate as to keep the temperature of the mixture at 10-15C.
After the addition was completed, ~he reactio~
mix~ure wa~ stirred for 40 minutes at 5-10.
Then most of the ethanol was removed under reduced pressure and the residue was dissolved in 130 ml. of water. The pH of the solution was lowered to 2.8 with 6 N hydrochloric acid and was maintained therP, while the mixture was extracted with 5 x 200 ml. of ethyl a etate.
The combined extracts were washed with ice water, dried over anhydrous magnesl.um sulfate, ~iltered and concen~rated under reduced ~ressure to an orange solid. This was weighed, redissolved.in 100 ml. of warm ethyl acetate and 1 equivalent (8.25 ml.~ 0.067 mole~) of dicyclohexylamine was added. Bright yellow crystals started to preclp-itate immediately. The mixture was cooled for 2 hours in an ice bath. Then the cry~tals were removed by filtrat~on,washed with ether,and dried ln vacuo yielding 14 ~. ~41% yield3. IR and NMR
~pectra~were consistent with structure.
Analysis:
Calcd. ~or C17H2~N20S2-1/2.H20: C, 58941; H,~ 8.35 , 8.02; S, 18.35.
Found: C, 58.~8; H, 8.22;
. N, 8.11; s,i 19.30.
5-méthylthiazole-2-~hiocarboxylic:acid dicycloh~xylam~ne ssle (13.5 g.) was slurried in ~06~7~ 1 150 ml, of water and layered wlth 150 ml. of ethyl acetate. The pH of the cooled solution was lowered to 2.2 with ~N hydrochloric acid. The layers were separated and the aqueous phase was extracted with 3 x 100 ml. of ethyl acetate. The combined extracts were dried over anhydrous magnesium sulfate, filtered and concentrated to a red solid whichr~as identified by infrared as thioacid;
weight 5~44 g.
To a stirred solution of 9.3 g. ~0O034 moles) of 7-aminocephalosporanic acid and 5.75 g. (0.068 moles) of sodium bicarbonate in 204 ml.: of aqueous phosphate buf~er at pH 6.4 was added 5.44 g. ~0.034 moIes) of 5-methyl~hiazole-2 thiocarboxylic acid.
The mixture was stirred in a nitrogen atmosphere at 50 for 5 hours keeping ~:he pH at 6.5 with 42% phosphoric acid. Then l:he mixture was cooled to 20 and the precipitated solid was removed by filtration, washed with water, flcetone and e~her and dried in vacuo over pho~3phorus pèntoxide to prov~de 4.4 g. (35% yiel~ of ~ ~an crystalline ~olid.~Infrared and N~ spectra were consistent with the structure.
Pivalo~loxymethyl 7-amino-3-~ me~hyl~iazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate Method A. The title compound is~produced by ~ubstituting for the 7-aminocephalosporanic .
acid used immediately above an equimolar weight -o pivaloyloxymethyl 7~aminocephalosporan~te hydrochloride prepared according to Example 2 of U.K. 1,229,453 from 7-aminocephalosporanic acid.~Germ~n 1,904,585 (Farmdoc 39,445~ is equivalent ~o U.K. 1,339,453.
~ '~ ; ..
` ` , ~ 45 -'~ .
Method B. The ~itle compound is produced .
by substituting for the 0.025 mole (6.8 g.) 7-aminocephalosporanic acid used in the pro-cedure of Example 2 of U.K. 1,229,453 an equi-molar weight of 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic .
acid. .
The respective acetoxymethyl, methoxy- .
methyl, acetonyl and phenacyl esters of 7-amino-3-(5-methylthiazol-2-yl)carb~nylthiomethyl-3- .
cephem-4-carboxylic acid are prepared by sub-8tituting in Method 8 above for the chloro-methyl pivalate used therein an equimolar weight of chloromethyl acetate, chloromethyl methyl .
ether, chloroacetone and phenacyl bromide, .
re~pectively.
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~ 7 ~ ~
3-Methyl-1,2~5-oxadiazole-4-thiolcarboxylic acid A mixture of 10.0 g. (7.8 mmole) of 3-methyl-1,2,5-oxadiazole-4-carboxylic acid, 40 mlO of thionyl chloride and 0.5 ml. of N,N-dimethyl-formamlde was heated under reflux for 66 hours.
The volatLle components were evaporated under reduced pressure ~t 35C./20 ~m" benzene was added to the residue and the ~ixture was re-evapor-ated. The oily acid chloride so obtained was dissolved in 10 ml. o~ dry benzene and the solu-~ion wàs added dropwise to a ~;tirred solution of 15~8 g. (17.2 mmole) of sodium sulfhydrate dihydrate ln 150 ml. of 90~/0 aqueous e~hanol 3t such ~ rate that the reaction temperature was in the range~
10-12C. After the addition was completed~ the slurry was stirred f~r 50 minutes at 10-15~C. nnd then f~ltered. The filtrate was evaporated under reduced pressure and the residue was dissolved ln 100 ml. of water. The pH of the solution was lowered ~o 2.5 with 6N hydrochloric acid~nd the product;was extracted into three 100 ml. portions of ether. The combined extracts were washed wi~h wa~er, dried over anhydrous magnesium sulfate ~nd evaporated to dryness under reduced pressure, The ~ '' 10 6'~7 0 1 resi(3ue was dis~illed to provide a yellow oilD b~p.
40~42C/0.8 mm, Yield 105 g. (13%~. .
7-Amino-3-(3-methyl-1,2~5-oxadiazol 4-yl)carbnYl- .
~_~ .
To a s~irred solut~on of 8.1 g. (29.8 mmole) o~ 7-aminocephalosporan~c acid and 5.0 g. (59.6 mmole) of sodium bicarbonate in 140 ml. of 0.1 M
~queous phosphate (buffered at pH 6.4) was added a solut~on of 4.3 g. (29.8 nnnolej of 3~methyl-1, 2,5-oxadiaæole-4-thiolcarboxylic acid in 15 mlO of acetone. The mixture was ætirred under a nitrogen ~tmosphere for 16 hours at 35C. The product was collecte~d by filtration, washed with water and .
acetone and dried in vacuo over phosphorus pentoxlde ~o afford 2.77 g. (26~/o) Of tan solid. ~
Analysis:~ .
Calcd.for: Cl2H~2N4o5s2 C9 40.45; H, 3.40;
N, 15.73.
, .
Found: C, 40.47; H, 3.50;
N~ 15.36.
. ,~ .
L~
.. .
4-yl)carbonylthiomethyl~3-ce~hem-4-carboxylate .
Me~hod A.
The title compound is produced by substituting Por the 7-aminocephalosporanic acid used immediately above an equimolar weight of pivaloyloxymethyl 7-aminocephalosporanate hydrochloride prepared ~L0~2701 accordlng to Example 2 of U.K. 1~229,453 from 7-ari~lnocephalosporanic acid. German ï 9 904,585 (Farmdoc 39,445) is equivalent to U.X. lj339,453.
Me~hod B.
The title compound is produced by substitutin~
for the 0.025 mole (6.8 g.) 7-aminocephalosporanic ~cid used in the procedure of Example 2 of U.K.
1,229,453 an equlmolar weight of 7-amino-3-(3-methyl-~ S-oxadiazol-4-yl)carbonylthiomethyl-3-cephem~
4-c~rboxylic acid.
The respective ace~oxymethyl, methoxymethyl, .
ace~ony~ and phenacyl esters of 7-amino-3-(3~methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid are prepared by substituting in Method B above for the chloromethyl pivalate used therein an equimolar weight of chloro~ethyl acetateJ
chlor~methyl methyl e~her, chloroacetone and phen~cyl bromide~, respectively.
' :
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~(~6~7~
EXAMPLES
. .
The following examples are given ~n illus~
tration of 9 but not in limitation of, the present invention.
~xample 1 :
7-~D~ a-Amino~henylacet~mido~-3-(thiazol-2- -~=~ L -.
CH-~-NH ~.S ~ ~
~H2 ~ ~ CH~-S-~ ~ S J
COO~ . -~8L-S567) T~ a stirred slurry of 4.5 g. (0.0126 m~le~) of 7-~mino-3-(thiazol-2 yl)carbonylthiomethyl-3~
cephem-4~carboxylic acid in 125 ml. of d~y methylene chloride were ~dded successively 3052 mI. tO.0252 moles): of triethylamine, 1.62 ml. (0.0126 moles) of N, N-dimethylaniline and 4.72 mlO (0~0378 moles) of trimethylchlorosilane. After being refluxed for 0.5 hour a clear brown solution resul~ed. It was cooled~to 0-5 and 2.B6 g. (0.0138 molea) of D~ 2-phenylglycyl chloride hydrochloride was added.
The resulting slurry w2s stirred for 2` h~oura letting the temperature rise to 25. This was then added to 125 ml. of ice water. The resulting tan precipi-~ate was s~irred for 15 minutes at room ~emperature~
filtered~ w~shed with water, acetone and eeher ~nd ~ried in vacuo over phosphorus pentoxide y~eldlng~3.8 g. of fl tan solid. The layerfi of the . . _ . . _ . . _. . ,, , ,, . , , j :
~L~6~0~
filtrate were separated and the aqueous phase was cooled, layered with 100 ml. of ethyl acetate and adjusted with 10%
sodium hydroxide to pH 4.2. The resulting white solid was removed by filtration, washad with water, acetone and ether and dried ln vacuo o~er phosphorous pentoxide to give 475 mg.
The 3.8 g. of the first precipitated solid was purified by slurrying in 50 ml. of water and 50 ml. of ethyl acetate and acidifying to pH 1.5 with 6 _ hydrochloric acid. It was stirred for 15 minutes and then filtered~ The filtrate was treated with 10~ sodium hydroxide to give pH 4.5. The solid which precipitated was collected by filtration, washed with water, acetone and ether and dried in vacuo over phosphorus pentoxide to give another 900 mg. Combined solids weighed 1.37 ~. (22%).
Infrared and NMR spectra were consistent with structure of the title product.
Analysis:
Calcd. for C20H18N4O5S3 H2O: C, 47.60; H, 3-98; N~ 11-03-Found: C, 47.40; H, 3.89; N, 10.79.
Example 2 .
Sodium 7-~D-t-)-~-amino~henylacetamido]-3-(thiazol-2-yl)-carbonylthiomethyl-3-cephem-4-carboxylate CH C NH ~ S
2 O N ~ 2 ~ S
COONa -To a stirred aqueous suspension of the zwitter-ionic form of 7-[D-(-)-~-aminophenylacetamido]-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4~carboxylic acid (0.8 mmole) is added lN aqueous NaOH at room temperature until a clear solution (pH 10.8) ,''.''''' ~
....... .. . .....
iZ701 i~ obtained. The solution is ~m~edifltely freeze-dried to give impurej solid sodium 7-[D~ a-amlnophenylDcetamido)-3-(thiazol-2-yl~)carbonyl-thiomethyl-3-cephem-4-carboxylate~
Following the same general procedure as abov.e, u~e of lN KOH in place of the lN NaOH used therein produces potassium 7- 1D-(-) -a-amlnophenylacetamldo]
-3-(t~iazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
~ .
7 ^ ~_ a _no -a- (3-thienyl)acetamido]-3-(thiazol-2-yl~c~rbonylthiomethyl-3-ce~hem-4-car xvlic acid r3 - CH-~-NH ~ 4 ~IN
COOH
The 3bove compound is prepared by s~bstituting an equ~molar weight o~ D-(-)-a-amino-a-(3-thienyl~
acetyl chlorlde hydrochloride in ~he procedure of : . Ex~mple 1 for the D-(-)-a-amino-~-phenylacetyl :~ chloride hydrochlorlde used ~herein.
: Example 4 a-amino-~-~2-thieny~2acetamido~-3-(_h~azol-I~CH-C NH
NH2 ~ ~ CH2~ S~ 11 S
The above compound is prepared by substituting ~n equimolar weight of D~ a-amino-a~(2-th~enyl) 49 ~ :
.
, ~0 6~7 ~ ~
acetyl chloride hydrochloride in the procedure of Example 1 for the D~ a-amino-~-phenylacetyl chloride hydrochloride used therein.
Example 5 Acetoxymethyl 7~ amino~henylacetamldo¦-3-H-~-NH ~ S
NH2 ~ ~ CH2-S-~
.
To a solution of acetoxymethyl 7-amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem-4-carboxylate (regenerated fxom 00009 mole of its hydrochloride) in 30 ml. ethyl acetate ~s added 0.020 mole pyr~d~ne. The ~ixture iB co~led in ice and s~irred while 0.010 mole D-~-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl ~cetate is added over ten minutes. After a further 20 minutes ` in the cold, stirring ~ s continued at room temperature for 1 hour. Then the mixture is washed successively wit~ aqueous NaHC03, 0.1 N HCl and water, dried ~nd evaporated in vacuo to leave the desired aeetoxymethyl 7 - 1D~ aminophenylacetamidol-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-c~rboxylate a~ ~n oll which crystallizes upon tritura~ion in cycl~hexane.
The re~pective pivaloyloxymethyl~ methoxy-methyl~, acetonyl and phenacyl esters corresponding .
..
627~L
to the absve acetoxymethyl ester are produced by replacing ~he acetoxymethyl 7-amino-3-(2-thiazolyl)-carbonylthiomethyl-3-cephem-4-carboxylate hydro-chloride used in the above procedure with 0.009 mole of the hydrochlor~de of pivaloyloxymethyl, methoxymethyl, acetonyl and phenacyl esters of 7-amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem-4-carboxylic acid, respectively.
F,xample 6 Acetoxymethyl 7-[D-a-amino-~-(3-thienyl~acetamido]-3-~thiazol-2-yl~carbonvlthiomethyl-3-ce~hem-4-carboxylate o CH-C-NH ~.S N
NH2 ~ N~ ~ CH2-S~
' ~OOCH20COCN3 . The above compound is prepared according to Example 5 by substituting for the D~ 2-phenyl-glycyl:chloxide hydrochloride used therein an equimolar amount of D-(-) -a-amino-a- (3~thienyl~
- ace~yl chloride hydrochloride.
~ The respective pivaloyloxymethyl, methoxy-methyl, acet~nyl and phenacyl esters corresponding - to the~:above acetoxymethyl ester are produced by replacing the acetoxymethyl 7-amino-3 ~2-thlazolyl)-c~rbonylthiomethyl-3-cephem-4-carboxylate hydro-chloride used in the ~bove procedure with 0.009 mole of the hydrochloride of plvaloyIoxymethyl, methoxymethyl, acetonyl and phenacyl~esters of 7-amLno~3~(2-thiazolyl~c~rbonyl~hiomethyl-3-cephem-4-carboxylic acid, respectively. `
.
~627 Example 7 A ~ o-a-~2-thienyl~acetamido~
-3-(thiaæol-2-yl)carbonylthlomethyl-3-cephem-4- -. .
carboxylate 1~ CH-C-IIH~S~ _~3 ; 2 ~ CH2-S-C
COOCH20COCH3' The above compound is prepared according to Example 5 by substituting for the D-(-)-2-phenyl-glycyl:chloride hydrochloride used there~n an equimolar amount of D-(-)-a-amino-a-(2-th~enyl) ace~yl chloride hydrochloriLde.
The respective pivaloyloxymethyl9 methoxy-methyl, acetonyl and phenac:yl esters corresponding to the above acetoxymethyl ester are produced by replacing the acetoxymethyl 7-amino-3-(2-thiazolyl)-carbonylthiome~hyl-3-cephem-4-carboxylate hydro-chloride used in ~he above procedure with 0.009 mole of the hydrochloride of pivaloyloxymethyl, methoxymethyl, ac,etonyl and phenacyl eseers of 7-amino-3-(2-thiazolyl~carbonylthiomethyl-3-: cephcm-4-carboxyllc acid, respectively.
:
.
, : - 52 -;~ , ~6Z~
Example 7- [D~ a-Aminophenylac2tamido 1-3- (5-methylthiazol-2-y~?carbonylthiomethyl-3-cephem-4-carboxylic acid o H_C_NH ~S ~ ~
NH2 ~ CH2_S-C ~ ~ H3 ~ ~ B s (BL-S586~ CO2H
To a ~tirred slurry of 4.4 g. (0.0119 mole~) of 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid in 120 ml. of dry methylene chloride were added successively 3.34 ml. (0.023 moles~ of triethylam~ne~ 1.51 ml. (0.011~ moles~ of M,N-dimèthylaniline and 4.45 ml. (0.0357 moles) of trimethylchloro~ilane. After being refluxed for 40 minutes, a clear yellow 801ution resulted. It was cooled to 3~ C. and 2.7 g. (0.0119 moles) of D~ 2-phenylgl:ycyl chloride hydrochloride was added with vigorous stirring. The resulting ~lurry was stirred for 2 hours letting the ~emperatur2 rise to 20. This was then added to 120 ml. o~ ice water and the resulting tan precipitate was stirred for 30 ~nutes ~t room temperature. It was filtered, washed wi~h wa~erj~acetone, and ether and.dried in vacuo over phosphorus pentoxidè yielding 4.1 g. of a tan solid.
The layers of the filtrate of the tan ~olid were ~epar~ted and the aqueous phase was cooled, layered with lOO ml. of ethyl acetate and adjustèd with 10%
sodium:hydroxide to pH 4.2. The resulting white solid was removed by filtration, washed with water, acetone; and ether and dried in vacuo~to give 75 mg, The 4.1 g. of the first precipit~ted solid was purifled by slurrying in 50 ml. of w~ter and 50 ml.
,.
~ 53 ~
~ ~ 6 ~ 7~ ~ :
of ethyl acetate and acidifyi~g to pH 1.5 wlth 6 N
hydrochloric acid. It was stirred for 15 minutes and the insoluble m~terial was filtered off. The filtrate was treated with 10% sodium hydroxide ~o give pH
4.5. The solid which precipitated was collected by filtration, washed with water, acetone3 and ether and dried in vacuo over phosphorus pentoxide to give 575 mgl Combined solids weighed 650 mg. ~ /o yield).
Infrared and NMR spectra were consistent with ~truc~ure.
Analysis:
Calcd. for C21~20N4OsS3 2H2o N 10 35' Found: C, 47.06; H, 4.97;
N, 9.35. :
Example 9 Sodium~7-[D~ aminophenylacetamido~-3-~5-methyl-thiazol-?.-yl~carbonylthiomethyl-3-cephem-4-carbo ylate H~_NH ~ S ~ ~
N~2 ~ ~ H2-S~ CH3 . CO~Na To a st~rred aqueous suspension of the zwitterionic for~ o~ 7- 1D~ a-aminophenylacetamido]-3-(5-methyl-thiazo~-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid (0.08 mmole) is added lN aqueous NaOH at room temperature until a clear solution ~pH 10.8) is obtsined.
The solution is lmmediately freeze-dried to give impure, solid sodium 7-~D~ -amlnophenylacetamido]-3-(5-methyl-thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
' . - 54 -,-- , , ;", ,- ; . I
, .. - . . _ . . . .
6Z7~1 Following the same general procedure a~ above~
use of lN KOH in pl~ce of the lN NaOH used therein produces potassium 7-[D~ a-aminophenylacetamido]-3-(S-methylthi~zol-2 yl)carbonylthiomethyl-3-cephem-4-carboxylate.
Example 10 acid ' -:
3 ~H ~ ~ ~ CU S C ~ ~ CU
The above compound is prepared by substituting an equimol~r weight of D-(~ amino ~-(3~thienyl)-acetyl chloride hydrochloride in the procedure of Example 8 for the D~(-)--2~phenylglycyl chloride hydrochloride used therein.
7-~D-~-amino-~-S2-thienyl)acetamido]-3~o(5-methyl ~cid H_C NH
2 O~ ~ CH2-~-p CH3 :~
CO~H
The ~bove compound is prepared by substltuting ~n equimo~r weight of D-(-)-a-amino-a-(2^thienyl)-;. . - 55 -acetyl chloride hydrochloride in the procedure of Example 8 for the D-(-)-.alpha.-amino-2-phenylglycyl chloride hydrochloride use therein.
Example 12 Acetoxymethyl 7-[D-(-)-.alpha.-aminophenylacetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
To a solution of acetoxymethyl 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate (regenerated from 0.009 mole of its hydrochloride) in 30 ml. ethyl acetate is added 0.002 mole pyridine. The mixture is cooled in ice and stirred while 0.010 mole D-(-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl acetate is added over ten minutes. After a further 20 minutes in the cold, stirring is continued at room temperature for 1 hour. Then the mixture is washed successively with aqueous NaHCO3, 0.1 N HCl and water, dried and evaporated in vacuo to leave the desired acetoxymethyl 7-[D-(-)-.alpha.-aminophenyl-acetamido[-3-(5-methlthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate as an oil which crystallizes upon trituration in cyclohexane.
The respective pivaloyloxymethyl, methoxymethyl, acetonyl and penacyl esters corresponding to the above acetoxymethyl ester are produced by replacing 3L~62~
the ~cetoxymethyl 7-amino-3- (5-methylthiazol-2-yl) -carbonylthiomethyl-3-cephem-4 carboxylate hydro-chloride used in the ~bove procedurc with O . 009 mole o~ the hydrochloride of pivaloyloxymethyl, methox~ethyl, ~cetonyl and phenacyl esters of 7-amino-3-~5-methylthiazol-2-yl~carbonylthiomethyl-3-cephem-4-carboxylic acid respeetively.
, Example 13 ~ -.
3-~5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate , H2-S~ SJ--CI~3 - ' . :
The above compound is prepared according to : Example 12 by ~ub~tituting for the D-(-)-2-phenyl- :-:~lycyl chloride hydrochloride used therein an equimolsr amount of D~ mino-a-(3-thienyl~
acetyl chloride hydrochlor~de.
: The~respective pivaloyloxymethylg me~hoxy-methy~ cetonyl and phenacyl esters cor~esponding to ~he~a~ove ~cetoxyme~hyl ~ster are produced.by repla~ing the acetoxymethyl 7-amlno-3-(5~methyl thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate hydrochloride used in the above procedure with 0.009 mole of the hydrochloride of plvaloyloxymethyl, methoxymethyl, ~cetonyl and phenacyl esters of . 7~mino-3-(5-me~hylthiazol-2-yl)carbonylthiomethyl-~ ;3-cephem-4-carboxylic acid, respectively~
.- ',:''^" ' . ........................... ' ' ..
1062~
Ex~mple 14 Acetoxymethyl 7-[D-a-amino-a-(2-thienYl)acetamidO1-3-(5-methylthiazol-2-yl?carbonylthiomethyl-3-cephem 4-c~rboxylate CH ~_NU S IN ~ C
O ~ CH2-S-~ ~ S H3 COOCH~OCOCH3 ., The above compound is prepared according to Example 12 by substitut:ing for the D~
2-phenylglycyl chloride hydrochioride used therein ~n equimolar amount of D-(-)-a-amino-a-(2.-thienyl)-acetyl chlor.ide hydrochloride.
The respective pivaloyloxymethyl, me~hoxy-methyl,.acetonyl and phenacyl esters correspondin~
to the~above acetoxymethyl ester are prep~red by r2pl~ing the acetoxymethyl 7-~mino-3~(5-methylthiazol- -2-yl)car~onyl~hiomethyl-3-cephem-4-carboxylate hydro-chloride used in the above procedure with 0.009 mole of the hydrochloride of pivaloyloxymethyl, methoxy-methyl, acetonyl and phenacyl esters ~of 7-amino-3-(S-methylthiazol-2-yl)carbonylthiome~hyl-3-cephem-4-carbaxylic acid, rcspectively.
Example 15 , ` The general procedure of Example 2 is repeated using the zwitterionic form~ of the acids shown below .
. 58 -:, , . ' , ': ' . , , . ,. , -~L~627~
in place of the 7 [D~ a-aminophenylacetamidol-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid ~sed therein.
' Use Of:
- 7-lD-a-amino--(3-thienyl)acetamido]-3-~thiazol-2-yl)carbonylthiQmethyl-3-cephem-4-carboxylic acid, 7-[D-a-amino-a-(2-thienyl)acetamido]-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, 7-[D-a-amino-a-(3-thienyl)acetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, and 7-[D-a-amino-a-(2-thienyl)acetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4'carboxylic acid produc,ed9 respectively, sodium 7-[D-~-amino-a-('3-thienyl)acetamido]- , ' 3-~thiaæol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate, pot~s~ium 7-~D-a-amino-a-(3-thienyl)acetamido]-3-(thia:zol-2-yl)carbonylthiomethyl-3-cephem-4- .
carboxylate, sod~ùm 7-lD-a-amino a-(2-thienyl~aeetamido~-3 , (thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carbox-ylate~ ~
.' potasslum 7-1D--amino~ 2-thienyl)acetamido]-, 3-(~hiazol-2-yl)carbonylthiomethyl-3-cephem-4-.;,., carboxylate, . sodi~um 7-[D-a-amino-~-(3-thienyl)a'cetamido]-.` ~3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-~.;.4-c~rboxyla~e~
: potassi~m 7-[D-a-smino-a-(3-thienyl)~ace~amido]-.-'.3-(5-me~hylthiazol-2-yl)carbonylthio~,ethyl-3-cephem-.`' 4-carboxylate, , .
....:~
. " _ 59 _ ...... .. _, .
.. . .. ..
3~06Z70~
sodium 7-1D-a-amino-a-(2-thienyl)acetamido]~
3-(5-methylthiazol-2-yl)carbonylthiomethyl 3-cephem-4-carboxylate, and potassium 7-[D-a-amino-a-(2-thienyl)acetamidol-3-(5-methylthlazol-2-yl)carbonylthiomethyl-3-cephem-4 c~rbcxylate.
.
~` -.
" ',~
. : ;:
- 60 ~
` ~ ~ 62 7 xample 16 7-[D-~ a-Aminophenylacetamido]-3-(3-me~hyl-l~2, 5 oxadiazol-4 -Yl? carbonYlthiomethYl-3-cePhem-4-carboxylic acid .
N
N~2 ~-S~
. ~ ...
To a stirred slurry of 2.70 g. (7.6 mmole) of 7-amlno-3-(3-methyl^l,2 9 5-oxadiazol-4~yl)carbonyl- .
thiome~hyl-3-cephem-4-carboxylic 3cid in lO0 ml.
of dry methylene chlor~de were added in succession 1~54 ~.;(15~2 mmole) of triethylam~ne~ 0.~92 g. ~7.6 mmole) of N,N-dLmethylanil~ne and 2.48~g. ~7.~ mmole) of trimethylchl~rosilane~ The mixture was heated .
under reflux for 20 minutes and the resùl~ing c~ear 60lutlon was cooled to ~O :In one poxtion, 1.57 g. ~:
~70~ mmole) o D~ 2-phenylglycyl chloride hydro-chloride was added and the mixture was stirred:vigorously for 1 hour at 2~5 and for l hour without ex~ern~l , . cooline.~ Addition of 50 ml. of water c~u~ed the , :.
., . ., . . , . .. , . , . . . . . .. : . . . .. 1 ~ ~Z 7 ~ ~
product to precipit~te and the solid was collected by f~ltration, washed with wster and acetone and dried in vacuo over phosphorus pen~oxide to provide 2.1 g.
(56%) of amorphous solid, mp 180-182C~ (decO). The infrared spectrum (KBr disc) showed absorption maxima (cm~l) at 1780 (~-lactam carbonyl)~ 1690 (amide carbonyl), 166n (thiolester carbonyl), 1620 and 1395 tcarboxylate) an~ 710 (phenyl~. The NMR
~pectrum of a solution in d6-dimethylsulfoxide and deuterium oxide gave signals (ppm from tetramethyl-~lane) which were assigned as follows: 7.47 (s, 5H) due to the benzene ring proton~:; 5.72 (d, lH) for the C7 proton; 5.03 ~s, lH) far the benzylic proton;
4.~7 (d,: lH) for the C6 proton~ 2.50 (s~ 3H) for the methyl protons; AB quartets centered at 4.15 and 3.50 ~or the exomethylene and C~ methylene protons respec-~ively.
This sample of 7-lD~ -aminophenylacetamido]-3-(3-methyl-1,2,5-oxadia~ol-4-yl)carbonylthiomethyl~
3-oephem-4-carboxyllc acid (called New Compound).after ~olution:in ~MS0 (dimethyl sulfoxide) at 14 mgm./ml.
followed by dilution with Nutrient Broth was found to exhibit the following Minimum Inhibitory Concen~
tr~tions (M.I.C.) in mcg./ml. versus the indicated mlcroorgani~sms as determined by overnight incub~tion at 37~C. by tube dilution. The in v~tro activity of cephalexin, a commercial o~al cephalosporin, is .
included in Table I.
... . . . ... . . . . ..
7~L
Table I
M. I . C in mc$ . /ml .
New Cepha- -~ ~ lexin D. pneumoniae ~ A9585 . ûl . 08 57O serum *
Str . pyogenes ~ A9604 . Ol ~ 08 5Z serum *
S, aureus Smith $ .A9537 . 6 . 6 S . aureus Smith + A9537 32 l. 3 50% serum $
S. aureus BX 1633-2 A9606 2 . 5 at lO-3 dilution S, aureu~ BX 1633-2 A961D6 8 2 st lO- dilu~ion S. aureus me~hi~illin- Al5097 32 16 resistant at 10-3 dilut ion S. aureus a'c 10~3 A97483 2 16 dilut io n S,. aureus at lO-2 A9748 32 ` 63 : dilution Sal, enteritidis ~ A9531 1 2 E~, coli Juhl ~ Al5119 32 8 ~:E. coll ~$ A~675 32 . ~ 16 X . pneumoniae ~ A9977 8 ~ ~ 4;. .
K. pneumon~iae $ Al5l30 63 ~ 16 Pr. miràbilis t A9900 4 4 Pr. mor~anii ~ A15153>125 ~125 Ps. aeruginosa ~: A9843A>125 .>125 `;
Ser. marceseens ~ ; A20019~l25 >l25 -.
* 5070 Nutrient Broth - 457O Antlbiotic Assay Bro~h at 104 dilution _ 63_ . .
' ~ 6iZ7~1 Blood levels in the mouse afoer oral admlnistration were determined with the following results: -Blood Level in mcg./ml.
R Dose 0.5 1 2 3.5 CH-C-NH~r-- ~ ~ mgm./kg. HrsO after adminis-NH~ ~ ~ ~ -CH2 - tral:iGn, ~ ~ .. ........ .. . --' , _ 100 ¦ 38.6 Z8.3 I3.7 4.4 ' -- ~ , loo 4104 26. 7 6 .o o . 87 cephalexin) monohydrate ..
.~ , , .
. ,. ? , ~. ' . i ' , , ,", . ~ ~ 9 . .
., , .~
'; ' ' ' ',: ,. . .
` _ '; ~', " ',- *,, ' ' :
64 ~
~ ,. ..; ..; .; ,,~,;,.. ,.,:...
' .
.... . . .. , .. ... ....... . . ..... . .. . .. " . . . . .. . .. .
-. :
1~627~1 Samples of the seven cephalosporin compoundc having the compound numbers reported above after ~ol~tion ln DMSO ~dimethyl sulfoxide) at 14 mgm./
ml. followed by dilution with Nutrient Broth were found to exhibit the following Minimum Inh~bitory Concentrations ~M.I.C.) ~n mcg./ml.
versus the indicated m~c~oorgan~sm~ as determined by overnight incubation ~t 37C. by Tube Dilution.
The in vitro activity of cephalexin W9S also tested.
~.
- : -',~`. ' ~: . ...
~ , : .
' . ' :
.
~L0627~:~
T~ble I
~i. 1. C . in mc~ . /ml .
Compound Cepha-r~anism No. 586 lexin D .pneumonia e ~ A9585 .16 . 6 5% serum *
Str . pyogenes + A9604 . 3 . 3 5/O serum *
S . aureus Smith $ A9537 . . 3 1. 3 S . aureus Smith + A9537 >63 2 . 5 50% s erurn ~: :
S . aureus PsX 1633-2 A9606 . 6 4 ~ t 10- 3 d ilution -S. allreus BX 1633-2 A96n6 4 8 at 10-2 dilution S. aureus methicil~in- A15t)97 4 32 resis~ant ~t 10- -dilution .
S. aureus at 10-3 A9748 8 32 dilution S. aureus~ at 10~ 2 A9748 63 l25 dilution Sal. enterltidis $ A9531 ,3 4 E. coli Juhl ~: A15119 2 8 E. coli ~ A9675 4 16 ~, pneum~ni~e $ A9977 4 8 K. pneumoniae ~ A15130 3~ :~ 16 ~ .
Pr . mirab~lis =~ A9900 16 ~ ~ 4 Pr. morganii ~ A15153:63. ~ 125 Ps . aeruginosa ~ A9843A>125 ~125 Ser. marcescens ~ A20019>125 ~ >125 .
* 50% Nutrient Broth _ 45D/o An~ibiotlc Ass~y Broth :~ ~t 10-~ dilu~ion ~ 30~
~L~6270~3L
Table II
M. I . C . in m~ . /ml .
Compound Cepha-Organism No. 567 lexin D . pneumoniae ~ A9585 . 04 5% s erum*
Str . pyogenes ~ A9604 . 04 . 3 5% ~erum *
S, aureu~ Smith $ A9537 ~3 o6 S. aureu~ Smith ~ A9537 8 2 5 50h ~erum 7 -S . aurèu~ BX 1633-2 A9606 . 6 2 ~t 10-~ dilution S. aureus BX 1633-2 A9606 . 2 4 at 10-2 dilution S. aureus methicillin- AlS097 4 32 res is tant a t 10-3 dilution S. aureus at 10-3 A9748 4 32 dilution S~ aureus :at 10-2 A9748 16 125 dilution Sal. enteritldis ~: A953i - .08 2 E. coli Juhl ~ A15119 1 ~ 8 E. coli ~:: A9675 b,~ lS
K. pneumoniae~ A9977 2 4 K. pneumoniae* A15130 8 16 ~' ,' . .
Pr. mirabilis ~: A9900 2 ~ 4 Pr. morganii $ A15153 63 ~ ~ ~125 Ps. aeruginosa ~ A9843A >125 ~125 Ser. marcescens ~: A20019 125 >125 * 50%~Nutrient Broth ~ 45% Antibiotic :Assay Broth :~ at 10-4 dilutLon Table III
M. I . C . in mcg . /ml .
CompoundCepha-~ No._591lexin D . pneumoniae ~ A95B5 . ~2 . 3 5VL serum *
Str . pyogenes ~ A9604 . 02 . 6 5% serum *
5. aureu~ Smith ~t A9537 .3 . 1.3 S. aureus Smil:h + A4537 8 2.5 50% serum *
. . .
S. aureus BX 1633-2 A9606 .6 4 ~t 10-3 dilution S. aureus BX 1633-2 A9606 2 4 at 10-Z dilution S. aureus methicillin- Al5lD97 2 63 resistant at 10-3 dilution S. aur~us:a~ 10-3 A9748 4 63 dilution S. aureus: ~t 10-~ A9748 16 125 dilution ~
Sal. enteritidis $ A9531 .16 4 1~. coli Juhl $ A15119 1 ~ ~ 8 E, coli ~ A9675 4 ~ 32 K. pneumoniae ~ A9977 ~ 1 4 K~ pneumoniae $ A15130 8~ ~ 32 Pr. mirabilis ~ A9900 ~ 4 Pr. morgani~ * A1515332 ~125 Ps. aeruginosa ~ A9843A>125 : ~125 Ser. marcescens :~ A20019125 ~ >125 * 50% Nutrient Broth ~ 45% Antib~otic As~ay Broth * a~ 10-4 dilution -.. . . . .. .. . ... . . . . .. . .... . . . . . . ... .
~L06Z7(;1~
Table IV
M I.C. in mcg./ml.
Compound Cepha- I
Or~nlsm No. 607 lexin D. pneumoniae ~ A9585 .16 .16 5% serum *
Str. pyogenes + A9604 .16 .16 5% ~erum *
S. aureus Smith ~ A9537 .3 o6 S. sureus Smlth + A9537 63 1.3 50% serum ~ I
S. aureus BX 1633-2 A9606 .6 2 1 .
~t 10-3 dilution S. aureua BX 1633-2 A9606 4 8 ~l ~t 10 ~ dilution S. aureus methlcil3in~ A15097 8 63 .
resistant a~ 10-dilution S. ~ureus at 10-3 A9748 8 63 .
dil~tion s. Aureus At 10 ? A9748 16 125 ~
dilution ~ . .
Sal. enter~tidis $ A9531 .6 4 .
: ~ E. coli Juhl $ A15119 8 . 8 .
E. coli . ~ ~ A9~75 16 ~ ~ 16 ~
K~ pneumoniae ~ A997716 .~ 4 .
: ~ pneumonia~.i A15130125 ~ ~ 16 .
Pr. mirabills $ A9900 `8 ~ ~ 8 Pr. morgan~ $ A151533~ : >125 p~. aerugiDosa $ A9843A~125 - >125 -Ser. marceScena ~ A20019~2$ ~125 * 50% Nutrient Bro~h - 45~ ~ntibiotic Assay Broth .
$~t 10-4 dilu~on - ~
~ ' . ' ,. ~ 33 ~1~627~
Table V
M.I.C. ~n mc~. /ml.
Compound Cephs-Or~anLsm No. 602_ lexin_ û ~ pneumon~ ae ~ A9585 . 60 3 5% serum *
S tr . pyogenes -~ A9604 ~ $ . 6 S% serum *
S. ~ureus Smith ~ A9537 ,3 1.3 S . aureus Smith + A9537 >63 2 . 5 50% ser~m ~
S . aureus BX 1633-2 A96061. 3 4 st 10-3 dilu~ion S. aureus BX 1633-2 A960616 at 10-2 dilution . S. aureus methicillin- A15097 4 32 resi~tant at 10-3 d llutlon S~ au~eus at 10-3A9748 8 32 ~ilut~on S. aureus at 10-2A9748 63 . 63 dilutiorl Sal. enteritidis ~ A9531 . 3 4 E. coli Juhl ~: A15119 ~ :~ 8 E. coli ~ ~ A9675 16 32 K. pneumonlàe ~ ~ A9977 16 8 K. pneumoniae ~ A1513032 ~ 16 Pr. mirabllis i A9900 16 8 Pr. morganii =~ A15153125. ~125 P8. aeruginos~ ~ A9843A>125 >125 .
Ser. marcescens $ A20019 ~12$ ~125 ~k 50% Nutrient Broth ~ 45% Antib~ot~c Ass~y Broth :
~ ~t 10-4 dilution ~ ¦
~ 34 -~a~6~7 Table VI
M . I . C . in mc ~ . /ml .
C:ompoundCepha-m No. 587lexin D O pneumon~ ae ~ Ag.~85 2 . 5 . 3 5% serum *
Str~ pyogenes + A9604 2 . 5 1. 3 S% serum *
S . aureu.s Sm~th ~ A9537 . 3 1. 3 S, aureus Smith ~ A9537 >63 2. 5 50% ~e~
S . aureus BX 1633-2 A9606 . 6 . 4 flt 10-3 dilut~on S. ~ureus BX 1633-2 A9606 1 4 ~t 10-2 dllutlon S. a~lreus methicillirl- Al5097 4 63 resistant a 10 3 dilut~on S. aur.eus at 10-3 A9748 4 63 dilution S. aureus at 10-2 A9748 1~: 125 dilution Sal. enterit~dis $ A9531 2 . 5 4 E. coli Juhl ~ A15119 I~ 8 E. coli ~ A9675 32 16 -K. pneumoniae =~ A997732 4 K. pneumoniae =~ A15130 ,~125 16 Pr . m~rabilis ~ A9900 >12 5~ ~ 4 Pr. morganii ~3: A15153 >125 >125 Ps. aeruginosa ~: A9843A ~125 . >125 Ser. marGes~cens ~ A20019 ,~125 ~ ~125 * 50/O Nutrient Broth - 45% Antibiotic As8ay Broth ~t 10-4 dilution :~CD62763:~L
Table VII
M~IoC~ in mcg./mlO
Compound Cepha-~ No. 556 lexin D. pneumoniae + A9585 1.3 .16 5% serum *
Str. pyogenes + A9604 2.5 .08 5% ~erum *
S. aureus Smith ~ A9537 1.3 .6 S. aureus Smith + A9537 63 1.3 50% s~rum ~ .
S. aureus BX 1633-2 A9606 1.3 2 ~t 10-3 dilution S. aureus BX 1633-2 A9606 4 4 at 10-2 dilution S. aureus methic~llin- A15097 4 32 resistant at 10-3 dilutlon S. aureùs ~t 10-3 A9748 ~ 32 : dilution s. sureus a~ 10-2 A9748 8 : 63 : : ~ dilution Sal. en~eritidis ~ A9531 ~ 2 E. coli 3uhl ~ ~ A15119 32 ~ 8 : E. coI~ $ A9~75 32 : 16 ~ .
K~ pneumoni~e ~ ~A9977 3~ 4 K. pneumoniae $: A15130 125 16 PL~. mirabllis =~ A99ûO 32 4 Pr . morganii ~ A15153 12 5 >125 ~s. aerug~nosa $ A9843A 125 >125 Ser. marceseens-~ A20019 63 >125 * 50~/~ Nutrient Broth - 45% Ant:~blot~c Assay Broth at 10-4 dllution :
1~6Z'7~
T~ble VIII
Blood levels in the mouse after oral admini~tration were determined with the following results:
. ~ , . .
- ._ Bloc d Level _in mc~. /ml.
fi~ S Dose O.5 1 2 3.5 ~CH-GONH--~ ~Hrg. aft2r ~dminis-I . l Imgm. /kg. tration.
~2 6 ~:H2R
.. . , ' ,. ~
-S-C ~H3 1.00 34 . 8 30 . 7 13 . 0 7 . O
., _ . .... ,._ , ., ' 11~3 100 l16.0:14.1 3.4<0.4 567 ~ : -. ~, ~ . ..
-s-~ ~
I ~ lo~ 6 s 7.9 4.4 3.2 591 : ~
_ .. . .... _~ - --~1CH3 ~ 1.2 1.4 1.3 1.0 , ~62~
Blood Level in ~g./ 1.
Dose O.S 1 2 3.5 Hrs. after adminis-m~m. /k~. tration _ ' ., . . ...
-sd~ oo Ill.7 17.6 9.7 3.3 602 . .
-S-d;~CH3 100 6.3 S.9 4.3 2.0 __ . . '':,~
I CH3 :.
-æ-~ ~sl 100 15.2 16.2 7.8<2.1 556 .
: _ . .
, . .~ .
-H 10044 . 6 ~ 3 . 4 6 . 1 1. 7 (cephalexin) ~onohydrate . -~:
~, ', ' ': : '' -.
.~ 8 ..
.... . , ... ... . . , . . . .... . . . . . _.. . ~ . .. ... .. . . . . .. . .
~627~
An examinat~on of Table VIII shows that very small struc~ural changes in the 3-acyl-thiomethyl substituent can greatly affect oral ~bsorption, The very closely-related compounds S86, 6~7 and 556,` for example, have blood levels after 3b minutes of 34.8, 1.2 and 15.2 mcg./ml.
An examination o~ Tables I - VII shows the same unpredictability with respect to in vitro activi-ties. Compounds 586 and 567 claimed in the present invention surprisingly and unexpectedly showed both excellent activity relative to cephalexin in the primary screening tests and good or-l absorption proper~ies.
.
. - , ' .~ . .
.
,, .
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~ . - 39 --~
i270 The novel medicaments provided by the present inven~on may be formulated as pharmaceutical com-posltions comprising, in addition to the active ingredient, a pharmaceu~ically acceptable carrier or diluent. The compounds may be administered both orally and parenterally. The pharmaceutical preparations may be in solid form such as capsules, ~ablets or dragees, or in liquid form such as solutlons, suspensions or emuls~ons. In the treatment of bacterial infections in man, the compounds of th~s invention may be administered in an amount of from about 5 to 200 mg./kg./day in divided dosage, e.g. 3 to 4 times a day.
They are administered in dosage units containing e.g. 125, 250 or 500 mg. of active ingredient with suitable physiologically ~ccep~able carriers or excipients.
In the experimental s'ections which follow, all temperatures unless indicated are in degrees Centigrade. The abbreviation 'tMeC12" is used to refer to methylene chloride while '~EA" is used to indicate triethylamine.
~' :
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, ~ :
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' 1~627~
PreE~ration Of Starting Materials Thiazole-2-thiocarboxylic acid dicyclohexylamine salt A mixture of 14.2 g. (0.11 moles) of 2-carboxy-thiazole, 220 ml. of methylene chloride, 13.9 g.
(0.11 moles) of oxalyl chloride and 0.5 ml. of N,N-dimethylformamide was stirred for 3 hours.
The insoluble material was removed by filtration, washed with methylene chlorid~ and the filtrate was evaporated under reduced pressure.to a solid.
This obtained acid chloride was-added portionwise to a st~rred and cooled solution of 24.2 g. (0,22 moles) o sodium sulfhydrate trihydrate in 247.5 ml.:of ethanol and 27.5 ml. of water ~t such a rate as to maintain the temperature of the reaction mixture at 10-15. After the addition was completed, the mixture was stirred for 40 minutes at 5-10.
Then the ethanol was removed under reduced pres~ure and the residue was dissolved in 145 ml. of water.
The pH o the solution was lowered to 2.8 by the addition of 6 N hydrochloric acid, and n~intained there:while the mixture was ex~racted w~*h 5 x 200 ml. of ethyl acetate. The eombined extracts were.washed with ice water, dried over:MgS04, filtered and evaporated under reduced pressure $o a reddish solid. This was weighe~d,-redissolved ;~.
in 100 ml. of warm ethyl acetate and l:equivalent :-.
of dicyclohexylsmine was added. Bright yellow crystals start2d to precipitate immediately.
The mixture was cooled in an ice-bath ~or 1 hour.
Then the crystals were removed by filtra~tion?
w~hed with e~her and dried under reduced.pressure .:
, - 41 -. .
~627~
for 15 hours; weight 15.2 g. (42.5% yield).
Analysis:
Calcd. for C16H26N20S2: C, 58.88; H, 8.04;
N, 8.62; S9 19.60.
Found: C~ 58.47; H, 8.11;
N~ 8.74; S, 19.91.
7-Amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem 4-carboxylic acid Fifteen g. of thiazole-2-thiocarboxylic acid dicyclohexylamine salt was ~lurried in 175 ml. of wa~er and layered with 175 ml. of ethyl acetate.
The pH of the cooled solution was lowered to 2.2 by the addition of 6 N hydrochloric acid. The layers were separated and the aqueous phase was extracked with 2 x 175 ml. of ethyl acetate. The combined extrac~s were driecl over MgS04, filtered and concentrated to a red solid, weight 5.35 g.
The infrared spectrum indicated thioacid.
To a ~tirred solution of 9.8 g. (0.036 moles) o 7-aminocephalosporanic acid and 6.05 g. (0.072 moles) o~ sodium bicarbonate in 170 ml. of aqueous phosphate buffer at pH 6.4 was added 5.35 g. (0.036 moles) of thia201e-2-thiocarboxylic acid. The mixture was stirred in a nitrogen a~mosphere at 50 for S hours keeping the pH at 6.6 with 42%
phosphoric acid. Then the mixture was cooled to 20 and t~e precipitated solid was ~emoved by filtration, washed with water, acetone and ether ~-~nd dried in _cuo over phosphorus pentoxide to provide 4,6 g. (35% yield) of a tan crystalline solid. Infrared ~nd NMR spectra were consistent with the structure.
, .
~ ~ ~Z 7~ ~
Pivaloyloxymethyl 7-amino-3-~2-thiazol~l)carbonyl-thiome~yl-3-cephem-4-carboxylate_ Method A. The title compound is produced by substituting for the 7-aminocephalosporanic acid used immediately above an equimolar weight of pivaloyloxymethyl 7-aminocephalosporanate hydrochloride prepared according to Example 2 of U.K. 1,229,453 from 7-aminocephalosporanic ~cid. German 1,904,585 (Farmdoc 39,445~ ls equLvalent to U.K. 1,3399453.
Method B. The title compound is produced by substituting for the 0.025 mole (6.8 g.) 7-aminocephalosporanic acid used in the pro-cedure of Example 2 o~ U.K. 1,229,453 an equi^
molar weight of 7-amino-3-t2-thiazolyl)carbonyl-thiomethyl-~-cephem-4-carboxylic acid.
The respective acetoxymethyl, methoxy-methyl, acetonyl and phenacyl esters o~ 7-amino-3-(2-th~azolyl)carbonylthiomethyl-3-cephem-4-carboxyl~c acid are prepared by substitut~ng in Method B above for the chloromethyl pivalate used therein an equimolar weight of chloromethyl acetate, chloromethyl methyl ether, chloroacetone and phenacyl bromide, respectively.
5-Methylthiazole-2-thiocarboxylic acid dicvclohexvlamine salt A mixture of 14.3 g. (O o l mole) of 5-methyl-thiazole-2-carboxylic ~cid in 200 ml.~of dry methylene` chloride, 12.6 g. (0.1 mole) of oxalyl chloride~and 0.5 ml. of N,~-dimethylformamide . .
was stirred for 3 hours at room temperature until a clear solution resulted. It was concentrated to a greenish oil, which was identified by in-frared as the acid chloride. This was added ~ ~3 -, Z7~
slowly over 5 minutes to a s~irred and cooled ~olution of 22.0 g. ~0~2 moles) of sodium sulf-hydrate trihydrate in 225 ml. of 100% ethanol and 25 ml. of water at such a rate as to keep the temperature of the mixture at 10-15C.
After the addition was completed, ~he reactio~
mix~ure wa~ stirred for 40 minutes at 5-10.
Then most of the ethanol was removed under reduced pressure and the residue was dissolved in 130 ml. of water. The pH of the solution was lowered to 2.8 with 6 N hydrochloric acid and was maintained therP, while the mixture was extracted with 5 x 200 ml. of ethyl a etate.
The combined extracts were washed with ice water, dried over anhydrous magnesl.um sulfate, ~iltered and concen~rated under reduced ~ressure to an orange solid. This was weighed, redissolved.in 100 ml. of warm ethyl acetate and 1 equivalent (8.25 ml.~ 0.067 mole~) of dicyclohexylamine was added. Bright yellow crystals started to preclp-itate immediately. The mixture was cooled for 2 hours in an ice bath. Then the cry~tals were removed by filtrat~on,washed with ether,and dried ln vacuo yielding 14 ~. ~41% yield3. IR and NMR
~pectra~were consistent with structure.
Analysis:
Calcd. ~or C17H2~N20S2-1/2.H20: C, 58941; H,~ 8.35 , 8.02; S, 18.35.
Found: C, 58.~8; H, 8.22;
. N, 8.11; s,i 19.30.
5-méthylthiazole-2-~hiocarboxylic:acid dicycloh~xylam~ne ssle (13.5 g.) was slurried in ~06~7~ 1 150 ml, of water and layered wlth 150 ml. of ethyl acetate. The pH of the cooled solution was lowered to 2.2 with ~N hydrochloric acid. The layers were separated and the aqueous phase was extracted with 3 x 100 ml. of ethyl acetate. The combined extracts were dried over anhydrous magnesium sulfate, filtered and concentrated to a red solid whichr~as identified by infrared as thioacid;
weight 5~44 g.
To a stirred solution of 9.3 g. ~0O034 moles) of 7-aminocephalosporanic acid and 5.75 g. (0.068 moles) of sodium bicarbonate in 204 ml.: of aqueous phosphate buf~er at pH 6.4 was added 5.44 g. ~0.034 moIes) of 5-methyl~hiazole-2 thiocarboxylic acid.
The mixture was stirred in a nitrogen atmosphere at 50 for 5 hours keeping ~:he pH at 6.5 with 42% phosphoric acid. Then l:he mixture was cooled to 20 and the precipitated solid was removed by filtration, washed with water, flcetone and e~her and dried in vacuo over pho~3phorus pèntoxide to prov~de 4.4 g. (35% yiel~ of ~ ~an crystalline ~olid.~Infrared and N~ spectra were consistent with the structure.
Pivalo~loxymethyl 7-amino-3-~ me~hyl~iazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate Method A. The title compound is~produced by ~ubstituting for the 7-aminocephalosporanic .
acid used immediately above an equimolar weight -o pivaloyloxymethyl 7~aminocephalosporan~te hydrochloride prepared according to Example 2 of U.K. 1,229,453 from 7-aminocephalosporanic acid.~Germ~n 1,904,585 (Farmdoc 39,445~ is equivalent ~o U.K. 1,339,453.
~ '~ ; ..
` ` , ~ 45 -'~ .
Method B. The ~itle compound is produced .
by substituting for the 0.025 mole (6.8 g.) 7-aminocephalosporanic acid used in the pro-cedure of Example 2 of U.K. 1,229,453 an equi-molar weight of 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic .
acid. .
The respective acetoxymethyl, methoxy- .
methyl, acetonyl and phenacyl esters of 7-amino-3-(5-methylthiazol-2-yl)carb~nylthiomethyl-3- .
cephem-4-carboxylic acid are prepared by sub-8tituting in Method 8 above for the chloro-methyl pivalate used therein an equimolar weight of chloromethyl acetate, chloromethyl methyl .
ether, chloroacetone and phenacyl bromide, .
re~pectively.
,, .' , ~ !
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.... ... ,. . .... .. _ . . . . . _ _ . , . . . ... . ._ . . . . . .
~ 7 ~ ~
3-Methyl-1,2~5-oxadiazole-4-thiolcarboxylic acid A mixture of 10.0 g. (7.8 mmole) of 3-methyl-1,2,5-oxadiazole-4-carboxylic acid, 40 mlO of thionyl chloride and 0.5 ml. of N,N-dimethyl-formamlde was heated under reflux for 66 hours.
The volatLle components were evaporated under reduced pressure ~t 35C./20 ~m" benzene was added to the residue and the ~ixture was re-evapor-ated. The oily acid chloride so obtained was dissolved in 10 ml. o~ dry benzene and the solu-~ion wàs added dropwise to a ~;tirred solution of 15~8 g. (17.2 mmole) of sodium sulfhydrate dihydrate ln 150 ml. of 90~/0 aqueous e~hanol 3t such ~ rate that the reaction temperature was in the range~
10-12C. After the addition was completed~ the slurry was stirred f~r 50 minutes at 10-15~C. nnd then f~ltered. The filtrate was evaporated under reduced pressure and the residue was dissolved ln 100 ml. of water. The pH of the solution was lowered ~o 2.5 with 6N hydrochloric acid~nd the product;was extracted into three 100 ml. portions of ether. The combined extracts were washed wi~h wa~er, dried over anhydrous magnesium sulfate ~nd evaporated to dryness under reduced pressure, The ~ '' 10 6'~7 0 1 resi(3ue was dis~illed to provide a yellow oilD b~p.
40~42C/0.8 mm, Yield 105 g. (13%~. .
7-Amino-3-(3-methyl-1,2~5-oxadiazol 4-yl)carbnYl- .
~_~ .
To a s~irred solut~on of 8.1 g. (29.8 mmole) o~ 7-aminocephalosporan~c acid and 5.0 g. (59.6 mmole) of sodium bicarbonate in 140 ml. of 0.1 M
~queous phosphate (buffered at pH 6.4) was added a solut~on of 4.3 g. (29.8 nnnolej of 3~methyl-1, 2,5-oxadiaæole-4-thiolcarboxylic acid in 15 mlO of acetone. The mixture was ætirred under a nitrogen ~tmosphere for 16 hours at 35C. The product was collecte~d by filtration, washed with water and .
acetone and dried in vacuo over phosphorus pentoxlde ~o afford 2.77 g. (26~/o) Of tan solid. ~
Analysis:~ .
Calcd.for: Cl2H~2N4o5s2 C9 40.45; H, 3.40;
N, 15.73.
, .
Found: C, 40.47; H, 3.50;
N~ 15.36.
. ,~ .
L~
.. .
4-yl)carbonylthiomethyl~3-ce~hem-4-carboxylate .
Me~hod A.
The title compound is produced by substituting Por the 7-aminocephalosporanic acid used immediately above an equimolar weight of pivaloyloxymethyl 7-aminocephalosporanate hydrochloride prepared ~L0~2701 accordlng to Example 2 of U.K. 1~229,453 from 7-ari~lnocephalosporanic acid. German ï 9 904,585 (Farmdoc 39,445) is equivalent to U.X. lj339,453.
Me~hod B.
The title compound is produced by substitutin~
for the 0.025 mole (6.8 g.) 7-aminocephalosporanic ~cid used in the procedure of Example 2 of U.K.
1,229,453 an equlmolar weight of 7-amino-3-(3-methyl-~ S-oxadiazol-4-yl)carbonylthiomethyl-3-cephem~
4-c~rboxylic acid.
The respective ace~oxymethyl, methoxymethyl, .
ace~ony~ and phenacyl esters of 7-amino-3-(3~methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid are prepared by substituting in Method B above for the chloromethyl pivalate used therein an equimolar weight of chloro~ethyl acetateJ
chlor~methyl methyl e~her, chloroacetone and phen~cyl bromide~, respectively.
' :
-''.
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~(~6~7~
EXAMPLES
. .
The following examples are given ~n illus~
tration of 9 but not in limitation of, the present invention.
~xample 1 :
7-~D~ a-Amino~henylacet~mido~-3-(thiazol-2- -~=~ L -.
CH-~-NH ~.S ~ ~
~H2 ~ ~ CH~-S-~ ~ S J
COO~ . -~8L-S567) T~ a stirred slurry of 4.5 g. (0.0126 m~le~) of 7-~mino-3-(thiazol-2 yl)carbonylthiomethyl-3~
cephem-4~carboxylic acid in 125 ml. of d~y methylene chloride were ~dded successively 3052 mI. tO.0252 moles): of triethylamine, 1.62 ml. (0.0126 moles) of N, N-dimethylaniline and 4.72 mlO (0~0378 moles) of trimethylchlorosilane. After being refluxed for 0.5 hour a clear brown solution resul~ed. It was cooled~to 0-5 and 2.B6 g. (0.0138 molea) of D~ 2-phenylglycyl chloride hydrochloride was added.
The resulting slurry w2s stirred for 2` h~oura letting the temperature rise to 25. This was then added to 125 ml. of ice water. The resulting tan precipi-~ate was s~irred for 15 minutes at room ~emperature~
filtered~ w~shed with water, acetone and eeher ~nd ~ried in vacuo over phosphorus pentoxide y~eldlng~3.8 g. of fl tan solid. The layerfi of the . . _ . . _ . . _. . ,, , ,, . , , j :
~L~6~0~
filtrate were separated and the aqueous phase was cooled, layered with 100 ml. of ethyl acetate and adjusted with 10%
sodium hydroxide to pH 4.2. The resulting white solid was removed by filtration, washad with water, acetone and ether and dried ln vacuo o~er phosphorous pentoxide to give 475 mg.
The 3.8 g. of the first precipitated solid was purified by slurrying in 50 ml. of water and 50 ml. of ethyl acetate and acidifying to pH 1.5 with 6 _ hydrochloric acid. It was stirred for 15 minutes and then filtered~ The filtrate was treated with 10~ sodium hydroxide to give pH 4.5. The solid which precipitated was collected by filtration, washed with water, acetone and ether and dried in vacuo over phosphorus pentoxide to give another 900 mg. Combined solids weighed 1.37 ~. (22%).
Infrared and NMR spectra were consistent with structure of the title product.
Analysis:
Calcd. for C20H18N4O5S3 H2O: C, 47.60; H, 3-98; N~ 11-03-Found: C, 47.40; H, 3.89; N, 10.79.
Example 2 .
Sodium 7-~D-t-)-~-amino~henylacetamido]-3-(thiazol-2-yl)-carbonylthiomethyl-3-cephem-4-carboxylate CH C NH ~ S
2 O N ~ 2 ~ S
COONa -To a stirred aqueous suspension of the zwitter-ionic form of 7-[D-(-)-~-aminophenylacetamido]-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4~carboxylic acid (0.8 mmole) is added lN aqueous NaOH at room temperature until a clear solution (pH 10.8) ,''.''''' ~
....... .. . .....
iZ701 i~ obtained. The solution is ~m~edifltely freeze-dried to give impurej solid sodium 7-[D~ a-amlnophenylDcetamido)-3-(thiazol-2-yl~)carbonyl-thiomethyl-3-cephem-4-carboxylate~
Following the same general procedure as abov.e, u~e of lN KOH in place of the lN NaOH used therein produces potassium 7- 1D-(-) -a-amlnophenylacetamldo]
-3-(t~iazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
~ .
7 ^ ~_ a _no -a- (3-thienyl)acetamido]-3-(thiazol-2-yl~c~rbonylthiomethyl-3-ce~hem-4-car xvlic acid r3 - CH-~-NH ~ 4 ~IN
COOH
The 3bove compound is prepared by s~bstituting an equ~molar weight o~ D-(-)-a-amino-a-(3-thienyl~
acetyl chlorlde hydrochloride in ~he procedure of : . Ex~mple 1 for the D-(-)-a-amino-~-phenylacetyl :~ chloride hydrochlorlde used ~herein.
: Example 4 a-amino-~-~2-thieny~2acetamido~-3-(_h~azol-I~CH-C NH
NH2 ~ ~ CH2~ S~ 11 S
The above compound is prepared by substituting ~n equimolar weight of D~ a-amino-a~(2-th~enyl) 49 ~ :
.
, ~0 6~7 ~ ~
acetyl chloride hydrochloride in the procedure of Example 1 for the D~ a-amino-~-phenylacetyl chloride hydrochloride used therein.
Example 5 Acetoxymethyl 7~ amino~henylacetamldo¦-3-H-~-NH ~ S
NH2 ~ ~ CH2-S-~
.
To a solution of acetoxymethyl 7-amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem-4-carboxylate (regenerated fxom 00009 mole of its hydrochloride) in 30 ml. ethyl acetate ~s added 0.020 mole pyr~d~ne. The ~ixture iB co~led in ice and s~irred while 0.010 mole D-~-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl ~cetate is added over ten minutes. After a further 20 minutes ` in the cold, stirring ~ s continued at room temperature for 1 hour. Then the mixture is washed successively wit~ aqueous NaHC03, 0.1 N HCl and water, dried ~nd evaporated in vacuo to leave the desired aeetoxymethyl 7 - 1D~ aminophenylacetamidol-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-c~rboxylate a~ ~n oll which crystallizes upon tritura~ion in cycl~hexane.
The re~pective pivaloyloxymethyl~ methoxy-methyl~, acetonyl and phenacyl esters corresponding .
..
627~L
to the absve acetoxymethyl ester are produced by replacing ~he acetoxymethyl 7-amino-3-(2-thiazolyl)-carbonylthiomethyl-3-cephem-4-carboxylate hydro-chloride used in the above procedure with 0.009 mole of the hydrochlor~de of pivaloyloxymethyl, methoxymethyl, acetonyl and phenacyl esters of 7-amino-3-(2-thiazolyl)carbonylthiomethyl-3-cephem-4-carboxylic acid, respectively.
F,xample 6 Acetoxymethyl 7-[D-a-amino-~-(3-thienyl~acetamido]-3-~thiazol-2-yl~carbonvlthiomethyl-3-ce~hem-4-carboxylate o CH-C-NH ~.S N
NH2 ~ N~ ~ CH2-S~
' ~OOCH20COCN3 . The above compound is prepared according to Example 5 by substituting for the D~ 2-phenyl-glycyl:chloxide hydrochloride used therein an equimolar amount of D-(-) -a-amino-a- (3~thienyl~
- ace~yl chloride hydrochloride.
~ The respective pivaloyloxymethyl, methoxy-methyl, acet~nyl and phenacyl esters corresponding - to the~:above acetoxymethyl ester are produced by replacing the acetoxymethyl 7-amino-3 ~2-thlazolyl)-c~rbonylthiomethyl-3-cephem-4-carboxylate hydro-chloride used in the ~bove procedure with 0.009 mole of the hydrochloride of plvaloyIoxymethyl, methoxymethyl, acetonyl and phenacyl~esters of 7-amLno~3~(2-thiazolyl~c~rbonyl~hiomethyl-3-cephem-4-carboxylic acid, respectively. `
.
~627 Example 7 A ~ o-a-~2-thienyl~acetamido~
-3-(thiaæol-2-yl)carbonylthlomethyl-3-cephem-4- -. .
carboxylate 1~ CH-C-IIH~S~ _~3 ; 2 ~ CH2-S-C
COOCH20COCH3' The above compound is prepared according to Example 5 by substituting for the D-(-)-2-phenyl-glycyl:chloride hydrochloride used there~n an equimolar amount of D-(-)-a-amino-a-(2-th~enyl) ace~yl chloride hydrochloriLde.
The respective pivaloyloxymethyl9 methoxy-methyl, acetonyl and phenac:yl esters corresponding to the above acetoxymethyl ester are produced by replacing the acetoxymethyl 7-amino-3-(2-thiazolyl)-carbonylthiome~hyl-3-cephem-4-carboxylate hydro-chloride used in ~he above procedure with 0.009 mole of the hydrochloride of pivaloyloxymethyl, methoxymethyl, ac,etonyl and phenacyl eseers of 7-amino-3-(2-thiazolyl~carbonylthiomethyl-3-: cephcm-4-carboxyllc acid, respectively.
:
.
, : - 52 -;~ , ~6Z~
Example 7- [D~ a-Aminophenylac2tamido 1-3- (5-methylthiazol-2-y~?carbonylthiomethyl-3-cephem-4-carboxylic acid o H_C_NH ~S ~ ~
NH2 ~ CH2_S-C ~ ~ H3 ~ ~ B s (BL-S586~ CO2H
To a ~tirred slurry of 4.4 g. (0.0119 mole~) of 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid in 120 ml. of dry methylene chloride were added successively 3.34 ml. (0.023 moles~ of triethylam~ne~ 1.51 ml. (0.011~ moles~ of M,N-dimèthylaniline and 4.45 ml. (0.0357 moles) of trimethylchloro~ilane. After being refluxed for 40 minutes, a clear yellow 801ution resulted. It was cooled to 3~ C. and 2.7 g. (0.0119 moles) of D~ 2-phenylgl:ycyl chloride hydrochloride was added with vigorous stirring. The resulting ~lurry was stirred for 2 hours letting the ~emperatur2 rise to 20. This was then added to 120 ml. o~ ice water and the resulting tan precipitate was stirred for 30 ~nutes ~t room temperature. It was filtered, washed wi~h wa~erj~acetone, and ether and.dried in vacuo over phosphorus pentoxidè yielding 4.1 g. of a tan solid.
The layers of the filtrate of the tan ~olid were ~epar~ted and the aqueous phase was cooled, layered with lOO ml. of ethyl acetate and adjustèd with 10%
sodium:hydroxide to pH 4.2. The resulting white solid was removed by filtration, washed with water, acetone; and ether and dried in vacuo~to give 75 mg, The 4.1 g. of the first precipit~ted solid was purifled by slurrying in 50 ml. of w~ter and 50 ml.
,.
~ 53 ~
~ ~ 6 ~ 7~ ~ :
of ethyl acetate and acidifyi~g to pH 1.5 wlth 6 N
hydrochloric acid. It was stirred for 15 minutes and the insoluble m~terial was filtered off. The filtrate was treated with 10% sodium hydroxide ~o give pH
4.5. The solid which precipitated was collected by filtration, washed with water, acetone3 and ether and dried in vacuo over phosphorus pentoxide to give 575 mgl Combined solids weighed 650 mg. ~ /o yield).
Infrared and NMR spectra were consistent with ~truc~ure.
Analysis:
Calcd. for C21~20N4OsS3 2H2o N 10 35' Found: C, 47.06; H, 4.97;
N, 9.35. :
Example 9 Sodium~7-[D~ aminophenylacetamido~-3-~5-methyl-thiazol-?.-yl~carbonylthiomethyl-3-cephem-4-carbo ylate H~_NH ~ S ~ ~
N~2 ~ ~ H2-S~ CH3 . CO~Na To a st~rred aqueous suspension of the zwitterionic for~ o~ 7- 1D~ a-aminophenylacetamido]-3-(5-methyl-thiazo~-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid (0.08 mmole) is added lN aqueous NaOH at room temperature until a clear solution ~pH 10.8) is obtsined.
The solution is lmmediately freeze-dried to give impure, solid sodium 7-~D~ -amlnophenylacetamido]-3-(5-methyl-thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
' . - 54 -,-- , , ;", ,- ; . I
, .. - . . _ . . . .
6Z7~1 Following the same general procedure a~ above~
use of lN KOH in pl~ce of the lN NaOH used therein produces potassium 7-[D~ a-aminophenylacetamido]-3-(S-methylthi~zol-2 yl)carbonylthiomethyl-3-cephem-4-carboxylate.
Example 10 acid ' -:
3 ~H ~ ~ ~ CU S C ~ ~ CU
The above compound is prepared by substituting an equimol~r weight of D-(~ amino ~-(3~thienyl)-acetyl chloride hydrochloride in the procedure of Example 8 for the D~(-)--2~phenylglycyl chloride hydrochloride used therein.
7-~D-~-amino-~-S2-thienyl)acetamido]-3~o(5-methyl ~cid H_C NH
2 O~ ~ CH2-~-p CH3 :~
CO~H
The ~bove compound is prepared by substltuting ~n equimo~r weight of D-(-)-a-amino-a-(2^thienyl)-;. . - 55 -acetyl chloride hydrochloride in the procedure of Example 8 for the D-(-)-.alpha.-amino-2-phenylglycyl chloride hydrochloride use therein.
Example 12 Acetoxymethyl 7-[D-(-)-.alpha.-aminophenylacetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate.
To a solution of acetoxymethyl 7-amino-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate (regenerated from 0.009 mole of its hydrochloride) in 30 ml. ethyl acetate is added 0.002 mole pyridine. The mixture is cooled in ice and stirred while 0.010 mole D-(-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl acetate is added over ten minutes. After a further 20 minutes in the cold, stirring is continued at room temperature for 1 hour. Then the mixture is washed successively with aqueous NaHCO3, 0.1 N HCl and water, dried and evaporated in vacuo to leave the desired acetoxymethyl 7-[D-(-)-.alpha.-aminophenyl-acetamido[-3-(5-methlthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate as an oil which crystallizes upon trituration in cyclohexane.
The respective pivaloyloxymethyl, methoxymethyl, acetonyl and penacyl esters corresponding to the above acetoxymethyl ester are produced by replacing 3L~62~
the ~cetoxymethyl 7-amino-3- (5-methylthiazol-2-yl) -carbonylthiomethyl-3-cephem-4 carboxylate hydro-chloride used in the ~bove procedurc with O . 009 mole o~ the hydrochloride of pivaloyloxymethyl, methox~ethyl, ~cetonyl and phenacyl esters of 7-amino-3-~5-methylthiazol-2-yl~carbonylthiomethyl-3-cephem-4-carboxylic acid respeetively.
, Example 13 ~ -.
3-~5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate , H2-S~ SJ--CI~3 - ' . :
The above compound is prepared according to : Example 12 by ~ub~tituting for the D-(-)-2-phenyl- :-:~lycyl chloride hydrochloride used therein an equimolsr amount of D~ mino-a-(3-thienyl~
acetyl chloride hydrochlor~de.
: The~respective pivaloyloxymethylg me~hoxy-methy~ cetonyl and phenacyl esters cor~esponding to ~he~a~ove ~cetoxyme~hyl ~ster are produced.by repla~ing the acetoxymethyl 7-amlno-3-(5~methyl thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate hydrochloride used in the above procedure with 0.009 mole of the hydrochloride of plvaloyloxymethyl, methoxymethyl, ~cetonyl and phenacyl esters of . 7~mino-3-(5-me~hylthiazol-2-yl)carbonylthiomethyl-~ ;3-cephem-4-carboxylic acid, respectively~
.- ',:''^" ' . ........................... ' ' ..
1062~
Ex~mple 14 Acetoxymethyl 7-[D-a-amino-a-(2-thienYl)acetamidO1-3-(5-methylthiazol-2-yl?carbonylthiomethyl-3-cephem 4-c~rboxylate CH ~_NU S IN ~ C
O ~ CH2-S-~ ~ S H3 COOCH~OCOCH3 ., The above compound is prepared according to Example 12 by substitut:ing for the D~
2-phenylglycyl chloride hydrochioride used therein ~n equimolar amount of D-(-)-a-amino-a-(2.-thienyl)-acetyl chlor.ide hydrochloride.
The respective pivaloyloxymethyl, me~hoxy-methyl,.acetonyl and phenacyl esters correspondin~
to the~above acetoxymethyl ester are prep~red by r2pl~ing the acetoxymethyl 7-~mino-3~(5-methylthiazol- -2-yl)car~onyl~hiomethyl-3-cephem-4-carboxylate hydro-chloride used in the above procedure with 0.009 mole of the hydrochloride of pivaloyloxymethyl, methoxy-methyl, acetonyl and phenacyl esters ~of 7-amino-3-(S-methylthiazol-2-yl)carbonylthiome~hyl-3-cephem-4-carbaxylic acid, rcspectively.
Example 15 , ` The general procedure of Example 2 is repeated using the zwitterionic form~ of the acids shown below .
. 58 -:, , . ' , ': ' . , , . ,. , -~L~627~
in place of the 7 [D~ a-aminophenylacetamidol-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid ~sed therein.
' Use Of:
- 7-lD-a-amino--(3-thienyl)acetamido]-3-~thiazol-2-yl)carbonylthiQmethyl-3-cephem-4-carboxylic acid, 7-[D-a-amino-a-(2-thienyl)acetamido]-3-(thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, 7-[D-a-amino-a-(3-thienyl)acetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, and 7-[D-a-amino-a-(2-thienyl)acetamido]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4'carboxylic acid produc,ed9 respectively, sodium 7-[D-~-amino-a-('3-thienyl)acetamido]- , ' 3-~thiaæol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylate, pot~s~ium 7-~D-a-amino-a-(3-thienyl)acetamido]-3-(thia:zol-2-yl)carbonylthiomethyl-3-cephem-4- .
carboxylate, sod~ùm 7-lD-a-amino a-(2-thienyl~aeetamido~-3 , (thiazol-2-yl)carbonylthiomethyl-3-cephem-4-carbox-ylate~ ~
.' potasslum 7-1D--amino~ 2-thienyl)acetamido]-, 3-(~hiazol-2-yl)carbonylthiomethyl-3-cephem-4-.;,., carboxylate, . sodi~um 7-[D-a-amino-~-(3-thienyl)a'cetamido]-.` ~3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-~.;.4-c~rboxyla~e~
: potassi~m 7-[D-a-smino-a-(3-thienyl)~ace~amido]-.-'.3-(5-me~hylthiazol-2-yl)carbonylthio~,ethyl-3-cephem-.`' 4-carboxylate, , .
....:~
. " _ 59 _ ...... .. _, .
.. . .. ..
3~06Z70~
sodium 7-1D-a-amino-a-(2-thienyl)acetamido]~
3-(5-methylthiazol-2-yl)carbonylthiomethyl 3-cephem-4-carboxylate, and potassium 7-[D-a-amino-a-(2-thienyl)acetamidol-3-(5-methylthlazol-2-yl)carbonylthiomethyl-3-cephem-4 c~rbcxylate.
.
~` -.
" ',~
. : ;:
- 60 ~
` ~ ~ 62 7 xample 16 7-[D-~ a-Aminophenylacetamido]-3-(3-me~hyl-l~2, 5 oxadiazol-4 -Yl? carbonYlthiomethYl-3-cePhem-4-carboxylic acid .
N
N~2 ~-S~
. ~ ...
To a stirred slurry of 2.70 g. (7.6 mmole) of 7-amlno-3-(3-methyl^l,2 9 5-oxadiazol-4~yl)carbonyl- .
thiome~hyl-3-cephem-4-carboxylic 3cid in lO0 ml.
of dry methylene chlor~de were added in succession 1~54 ~.;(15~2 mmole) of triethylam~ne~ 0.~92 g. ~7.6 mmole) of N,N-dLmethylanil~ne and 2.48~g. ~7.~ mmole) of trimethylchl~rosilane~ The mixture was heated .
under reflux for 20 minutes and the resùl~ing c~ear 60lutlon was cooled to ~O :In one poxtion, 1.57 g. ~:
~70~ mmole) o D~ 2-phenylglycyl chloride hydro-chloride was added and the mixture was stirred:vigorously for 1 hour at 2~5 and for l hour without ex~ern~l , . cooline.~ Addition of 50 ml. of water c~u~ed the , :.
., . ., . . , . .. , . , . . . . . .. : . . . .. 1 ~ ~Z 7 ~ ~
product to precipit~te and the solid was collected by f~ltration, washed with wster and acetone and dried in vacuo over phosphorus pen~oxide to provide 2.1 g.
(56%) of amorphous solid, mp 180-182C~ (decO). The infrared spectrum (KBr disc) showed absorption maxima (cm~l) at 1780 (~-lactam carbonyl)~ 1690 (amide carbonyl), 166n (thiolester carbonyl), 1620 and 1395 tcarboxylate) an~ 710 (phenyl~. The NMR
~pectrum of a solution in d6-dimethylsulfoxide and deuterium oxide gave signals (ppm from tetramethyl-~lane) which were assigned as follows: 7.47 (s, 5H) due to the benzene ring proton~:; 5.72 (d, lH) for the C7 proton; 5.03 ~s, lH) far the benzylic proton;
4.~7 (d,: lH) for the C6 proton~ 2.50 (s~ 3H) for the methyl protons; AB quartets centered at 4.15 and 3.50 ~or the exomethylene and C~ methylene protons respec-~ively.
This sample of 7-lD~ -aminophenylacetamido]-3-(3-methyl-1,2,5-oxadia~ol-4-yl)carbonylthiomethyl~
3-oephem-4-carboxyllc acid (called New Compound).after ~olution:in ~MS0 (dimethyl sulfoxide) at 14 mgm./ml.
followed by dilution with Nutrient Broth was found to exhibit the following Minimum Inhibitory Concen~
tr~tions (M.I.C.) in mcg./ml. versus the indicated mlcroorgani~sms as determined by overnight incub~tion at 37~C. by tube dilution. The in v~tro activity of cephalexin, a commercial o~al cephalosporin, is .
included in Table I.
... . . . ... . . . . ..
7~L
Table I
M. I . C in mc$ . /ml .
New Cepha- -~ ~ lexin D. pneumoniae ~ A9585 . ûl . 08 57O serum *
Str . pyogenes ~ A9604 . Ol ~ 08 5Z serum *
S, aureus Smith $ .A9537 . 6 . 6 S . aureus Smith + A9537 32 l. 3 50% serum $
S. aureus BX 1633-2 A9606 2 . 5 at lO-3 dilution S, aureu~ BX 1633-2 A961D6 8 2 st lO- dilu~ion S. aureus me~hi~illin- Al5097 32 16 resistant at 10-3 dilut ion S. aureus a'c 10~3 A97483 2 16 dilut io n S,. aureus at lO-2 A9748 32 ` 63 : dilution Sal, enteritidis ~ A9531 1 2 E~, coli Juhl ~ Al5119 32 8 ~:E. coll ~$ A~675 32 . ~ 16 X . pneumoniae ~ A9977 8 ~ ~ 4;. .
K. pneumon~iae $ Al5l30 63 ~ 16 Pr. miràbilis t A9900 4 4 Pr. mor~anii ~ A15153>125 ~125 Ps. aeruginosa ~: A9843A>125 .>125 `;
Ser. marceseens ~ ; A20019~l25 >l25 -.
* 5070 Nutrient Broth - 457O Antlbiotic Assay Bro~h at 104 dilution _ 63_ . .
' ~ 6iZ7~1 Blood levels in the mouse afoer oral admlnistration were determined with the following results: -Blood Level in mcg./ml.
R Dose 0.5 1 2 3.5 CH-C-NH~r-- ~ ~ mgm./kg. HrsO after adminis-NH~ ~ ~ ~ -CH2 - tral:iGn, ~ ~ .. ........ .. . --' , _ 100 ¦ 38.6 Z8.3 I3.7 4.4 ' -- ~ , loo 4104 26. 7 6 .o o . 87 cephalexin) monohydrate ..
.~ , , .
. ,. ? , ~. ' . i ' , , ,", . ~ ~ 9 . .
., , .~
'; ' ' ' ',: ,. . .
` _ '; ~', " ',- *,, ' ' :
64 ~
~ ,. ..; ..; .; ,,~,;,.. ,.,:...
6~ 7 __ 7 ~ ,.
~CH ~ H ~ S~
NH2 o~N ~H2-S-~
To a stirred aqueous suspension of the zwitter~
ionic form of 7-lD~ a~aminophenylace~am~do]-3~
~3 methyl-19~,5-oxadiazol-4~yl)carbonylthiomethyl-3 cephem~4-carboxylic acid (0.8 mmole) is added lN aqueous NaOH at room temperature untll a clear solution (pH 10.8) is ob~ained. The solution is ~mmediately freeze-dried to give impure, solLd sodium 7~[D~ aminophenylacetamido]-3-(3-methyl-1,~,5-oxadiazol-4 yl)carbonylthiomethyl-3-cephem-4~carboxylate.
Following ~he same general procedure as above, use of lN XOH in place of the NaOH used therein produces potass~um 7 lD-(-3-~-aminophenylacetamido]-3~t3-methyl-1?2,5-oxadlazol-4-yl)carbonylthiomethyl-3-cephem-4~carboxylate~ . .
- ~5 ~ .
t ~ ~r~ r
~CH ~ H ~ S~
NH2 o~N ~H2-S-~
To a stirred aqueous suspension of the zwitter~
ionic form of 7-lD~ a~aminophenylace~am~do]-3~
~3 methyl-19~,5-oxadiazol-4~yl)carbonylthiomethyl-3 cephem~4-carboxylic acid (0.8 mmole) is added lN aqueous NaOH at room temperature untll a clear solution (pH 10.8) is ob~ained. The solution is ~mmediately freeze-dried to give impure, solLd sodium 7~[D~ aminophenylacetamido]-3-(3-methyl-1,~,5-oxadiazol-4 yl)carbonylthiomethyl-3-cephem-4~carboxylate.
Following ~he same general procedure as above, use of lN XOH in place of the NaOH used therein produces potass~um 7 lD-(-3-~-aminophenylacetamido]-3~t3-methyl-1?2,5-oxadlazol-4-yl)carbonylthiomethyl-3-cephem-4~carboxylate~ . .
- ~5 ~ .
t ~ ~r~ r
7~
7 --1D~ a-Amino-a- (3-thienyl)acetamidol-3-(3-methyl-1~2,5-oxadiazol-4-Yl~arbon~lthiomethyl-3-cephem-4-c~rboxylic acid h_C_NH
NH2 N H~-S-~
COOH
The above compound ~s prepared by substitut~ng an equimolar weigh~ of D~ mino-~-(3-thienyl)-~cetyl chlorlde hydrochloride in the procedure of Examplel~ for the D-(-)-2-phenylglycyl chloride hydrochloride used therein Example19 ~rbollvlic ~-id, .
~H-C-NH ~ S ~ C ~
~ CH~ S ~ ¦
The ~bove compound is prepared by ~obstituting an ~quimolar weight of D-(-)-a~amino~a-(~2-thienylj-acetyl chloride hydrochlor~de in the procedure of Example 16 for the D-(~)-2~phenylglycyl chloride hydrochloride used therein. :
:~ 66 ~ `
3L~62 ~e~Q
A ~ ~ b~5~5~L~ooa~2 3-cephem-4-carboxylate ~IH-RC-~ L R
NH2 ~ CH2-S-C ~ ~
To a ~olution of acetoxymethyl 7-amino-3-~3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl~
3-cephem-4-carboxylate (regenerated from 0.009 mole of l~s hydrochlor~de) in 30 ml. ethyl acet~te is added 0.020 mole pyridine. Th~ mixture is cooled in ice and st~rred while 0.010 mole D-(-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl acetate i3 ~dded over ten minu~es. After a furtller 20 m~nute~ in the cold, stirring ls continued at rooD
temperature ~or 1 hour. Then the mixture is washed success~vely with aqueous NaHC03, O.lN HCl and water, drled ~nd evaporated in vacuo to leave ~he desired ~cetoxy~ethyl 7-[D~ aminophenyl-sce~amido3~3-~3-methyl-1,2 5-oxadia2O1-4-yl)carbonyl- ;
thiomethyl-3-cephem-4-carboxylate as an oil which crystallizes upon trltura~ion ln cyclohexane, The re~pec~ive pivaloyloxymethyl~ methoxy-methyl, acetonyl and phenacyl esters correspondlng ~ 7 ~6Z7~1 to the above acetoxymethyl es~er are produced by repl~cing the acetoxymethyl 7 amino-3-(3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxy-late hydrochloride used in the above procedure with 0.009 mole of the hydrochloride of piv~loyloxymethyl~ methoxymethyl, acetonyl and phenacyl esters of 7 amino-3-(3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiome~hyl-3-cephem-4_ carboxylic ecid, respectively.
Example2,-1 Acetoxymethyl 7-[D~ -amino-~-(3-thien~l)-acet~m~do~-3-(3-methy~ 2,5-o:Kadiazol-4-Yl)carbonyl thiome~hyl-3-cephem-4-carboxylate .
~H2 ,~Cd2-s R~I
The above compound is prepared according to Example aPby substituting for the D-(-)-2-phenylglycyi chloride hydrochloride used therein an equimolar amo~mt of D~ -amlno~:a-(3-thienyl)acetyl chloride hydrochloride. ~ ~
The respective pivaloyloxymethyl, me~hoxymethyl, ~cetonyl;and phenaeyl esters correspondlng to the above ocetDxymethyl e~ter ~re produced by replacing the acetoxymethyl 7~amino-3-(3-methyl-1,2,5-ox~diazol - ~8 , - - -- ~ \
~ 6 Z7 ~ ~
4-yl)carbonyl~hiomethyl-3-cephem-4~carboxylate hydrochloride used ln the above procedure with 0.009 mole of the hydrochloride of pivaloyloxy-methyl, methoxymethyl, acetonyl and phenacyl ester~i of 7-amino-3-(3-methyl-1,2 9 5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, respectively.
-Acetoxymethyl 7-[D-(-)-a-amino ~ -thienyl)-acetamldo]-3~(3-methyl-1,2,5-oxadiazol-4-yl)carbonyl-thiometh~l-3-cephem-4-carboxylate .
NH ~ ~ ~ ~ N
2 ~ N ~ CH2-S~
COOCH~OCOCN3 The a~ove compound is prepared accord~ng to Example2~ by su~stituting for the D~ 2-phenyl-glycyl chlorlde hydrochloride used therein an equimolar amount o~ D~ -amlno~-(2~th1enyl3-~cetyl chloride hydrochloride.
The respeetiYe pivaloyloxyme~hyl, methoxy~
methyl, acetonyl and phenacyl esters corresponding to the above ~cetoxyme~hyl ester are produced by replncing the acetoxymethyl 7~amino-3-(3-me~hyl-1, 2, 5 -oxadlazol ~4-y~) carbonylthiomethyl-3-c ephem-4-carbo~ylate hydrochloride used ~ n the above procedure with 0.009 mole of the hydrochloride of .
_ 6~ _ ~ ~ 6Z 7~ ~
pivaloyloxymethyl, methoxymethyl, ~cetonyl snd phenaeyl esters of 7-amino-3-~3-methyl-1,295-oxadiazol-4-yl)carbonylthiomethyl-3-cephem~4-c~rboxylic acid, respectively.
Example 2-3 The general procedure of Example 1~ is repeated using the zwitterionic forms of the acids shown below in place of the 7-~D~ a-aminophenylacetamido~
3~3-methyl-1,2,5-oxadiazol-4-yl~earbonylthiomethyl-3-cephem-4-carboxylic acid used therein.
Use of:
7-[D~ a-amino-a-(3-thienyl)acetamido]-3-(3-me~hyl-1,2 9 5-oxadiazol-4-yl)carbonyl~hiomethyl-3-cephem-4-carboxyl~c acid, and 7 - [D~ a~amino-a- (2-~hienyl)acetamido]-3-(3-methyl-1,2,5-oxadia~ol-4-yl)carbcnylthiomethyl-3-cephem-4-carboxylic acid produeed, respectively, Sodi~m 7 lD~ a-amino-a-(3-thienyl)acetam~dol^
3-(3~methyl-l,2,5-ox~d~szol-4-yl)carbonylthiomethyl- -3-cephem-4-carboxylate, Potassium 7-[D~ a-amino-a-(3-thieny~)ace~amido3 3- (3-methyl-1, 2, 5 -oxadlazol~4-yl) carbonylthiomethyl-3 -cephem~4-carboxylate, Sodium ~[D-(-) Da-amino ~a- (2-thienyl)~acetamido]- -3-(3-methyl-192,5-oxad~azol-4-yl~carbonylthiomethyl~
3~cephem-4~carboxylate, ~nd . ~ , . - ' .
_ 70 , ~06;~70~
Potassium 7-[D-~ ~-a-amino-a-(2-th~enyl)-~cetamido]-3-(3-methyl-1,2,5-oxadiazol-4-yl)-carbonylthLomethyl-3-cephem-4-carboxylate.
' ` .
: - :
: :
.
- -:
, .
7 --1D~ a-Amino-a- (3-thienyl)acetamidol-3-(3-methyl-1~2,5-oxadiazol-4-Yl~arbon~lthiomethyl-3-cephem-4-c~rboxylic acid h_C_NH
NH2 N H~-S-~
COOH
The above compound ~s prepared by substitut~ng an equimolar weigh~ of D~ mino-~-(3-thienyl)-~cetyl chlorlde hydrochloride in the procedure of Examplel~ for the D-(-)-2-phenylglycyl chloride hydrochloride used therein Example19 ~rbollvlic ~-id, .
~H-C-NH ~ S ~ C ~
~ CH~ S ~ ¦
The ~bove compound is prepared by ~obstituting an ~quimolar weight of D-(-)-a~amino~a-(~2-thienylj-acetyl chloride hydrochlor~de in the procedure of Example 16 for the D-(~)-2~phenylglycyl chloride hydrochloride used therein. :
:~ 66 ~ `
3L~62 ~e~Q
A ~ ~ b~5~5~L~ooa~2 3-cephem-4-carboxylate ~IH-RC-~ L R
NH2 ~ CH2-S-C ~ ~
To a ~olution of acetoxymethyl 7-amino-3-~3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl~
3-cephem-4-carboxylate (regenerated from 0.009 mole of l~s hydrochlor~de) in 30 ml. ethyl acet~te is added 0.020 mole pyridine. Th~ mixture is cooled in ice and st~rred while 0.010 mole D-(-)-2-phenyl-glycyl chloride hydrochloride in 30 ml. ethyl acetate i3 ~dded over ten minu~es. After a furtller 20 m~nute~ in the cold, stirring ls continued at rooD
temperature ~or 1 hour. Then the mixture is washed success~vely with aqueous NaHC03, O.lN HCl and water, drled ~nd evaporated in vacuo to leave ~he desired ~cetoxy~ethyl 7-[D~ aminophenyl-sce~amido3~3-~3-methyl-1,2 5-oxadia2O1-4-yl)carbonyl- ;
thiomethyl-3-cephem-4-carboxylate as an oil which crystallizes upon trltura~ion ln cyclohexane, The re~pec~ive pivaloyloxymethyl~ methoxy-methyl, acetonyl and phenacyl esters correspondlng ~ 7 ~6Z7~1 to the above acetoxymethyl es~er are produced by repl~cing the acetoxymethyl 7 amino-3-(3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxy-late hydrochloride used in the above procedure with 0.009 mole of the hydrochloride of piv~loyloxymethyl~ methoxymethyl, acetonyl and phenacyl esters of 7 amino-3-(3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiome~hyl-3-cephem-4_ carboxylic ecid, respectively.
Example2,-1 Acetoxymethyl 7-[D~ -amino-~-(3-thien~l)-acet~m~do~-3-(3-methy~ 2,5-o:Kadiazol-4-Yl)carbonyl thiome~hyl-3-cephem-4-carboxylate .
~H2 ,~Cd2-s R~I
The above compound is prepared according to Example aPby substituting for the D-(-)-2-phenylglycyi chloride hydrochloride used therein an equimolar amo~mt of D~ -amlno~:a-(3-thienyl)acetyl chloride hydrochloride. ~ ~
The respective pivaloyloxymethyl, me~hoxymethyl, ~cetonyl;and phenaeyl esters correspondlng to the above ocetDxymethyl e~ter ~re produced by replacing the acetoxymethyl 7~amino-3-(3-methyl-1,2,5-ox~diazol - ~8 , - - -- ~ \
~ 6 Z7 ~ ~
4-yl)carbonyl~hiomethyl-3-cephem-4~carboxylate hydrochloride used ln the above procedure with 0.009 mole of the hydrochloride of pivaloyloxy-methyl, methoxymethyl, acetonyl and phenacyl ester~i of 7-amino-3-(3-methyl-1,2 9 5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxylic acid, respectively.
-Acetoxymethyl 7-[D-(-)-a-amino ~ -thienyl)-acetamldo]-3~(3-methyl-1,2,5-oxadiazol-4-yl)carbonyl-thiometh~l-3-cephem-4-carboxylate .
NH ~ ~ ~ ~ N
2 ~ N ~ CH2-S~
COOCH~OCOCN3 The a~ove compound is prepared accord~ng to Example2~ by su~stituting for the D~ 2-phenyl-glycyl chlorlde hydrochloride used therein an equimolar amount o~ D~ -amlno~-(2~th1enyl3-~cetyl chloride hydrochloride.
The respeetiYe pivaloyloxyme~hyl, methoxy~
methyl, acetonyl and phenacyl esters corresponding to the above ~cetoxyme~hyl ester are produced by replncing the acetoxymethyl 7~amino-3-(3-me~hyl-1, 2, 5 -oxadlazol ~4-y~) carbonylthiomethyl-3-c ephem-4-carbo~ylate hydrochloride used ~ n the above procedure with 0.009 mole of the hydrochloride of .
_ 6~ _ ~ ~ 6Z 7~ ~
pivaloyloxymethyl, methoxymethyl, ~cetonyl snd phenaeyl esters of 7-amino-3-~3-methyl-1,295-oxadiazol-4-yl)carbonylthiomethyl-3-cephem~4-c~rboxylic acid, respectively.
Example 2-3 The general procedure of Example 1~ is repeated using the zwitterionic forms of the acids shown below in place of the 7-~D~ a-aminophenylacetamido~
3~3-methyl-1,2,5-oxadiazol-4-yl~earbonylthiomethyl-3-cephem-4-carboxylic acid used therein.
Use of:
7-[D~ a-amino-a-(3-thienyl)acetamido]-3-(3-me~hyl-1,2 9 5-oxadiazol-4-yl)carbonyl~hiomethyl-3-cephem-4-carboxyl~c acid, and 7 - [D~ a~amino-a- (2-~hienyl)acetamido]-3-(3-methyl-1,2,5-oxadia~ol-4-yl)carbcnylthiomethyl-3-cephem-4-carboxylic acid produeed, respectively, Sodi~m 7 lD~ a-amino-a-(3-thienyl)acetam~dol^
3-(3~methyl-l,2,5-ox~d~szol-4-yl)carbonylthiomethyl- -3-cephem-4-carboxylate, Potassium 7-[D~ a-amino-a-(3-thieny~)ace~amido3 3- (3-methyl-1, 2, 5 -oxadlazol~4-yl) carbonylthiomethyl-3 -cephem~4-carboxylate, Sodium ~[D-(-) Da-amino ~a- (2-thienyl)~acetamido]- -3-(3-methyl-192,5-oxad~azol-4-yl~carbonylthiomethyl~
3~cephem-4~carboxylate, ~nd . ~ , . - ' .
_ 70 , ~06;~70~
Potassium 7-[D-~ ~-a-amino-a-(2-th~enyl)-~cetamido]-3-(3-methyl-1,2,5-oxadiazol-4-yl)-carbonylthLomethyl-3-cephem-4-carboxylate.
' ` .
: - :
: :
.
- -:
, .
Claims (22)
1. A process for the preparation of a compound having the D-configuration in the 7-sidechain and having the formula (I) wherein R is hydrogen, pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl; and R' is 2-thiazolyl or 5-methyl-2-thiazolyl or 3-methyl-1,2,5-oxadiazol-4-yl; or a pharmaceutically acceptable salt thereof, comprising:
(a) when the starting material includes the desired final substituent in the 7-position of the cephem nucleus:
reacting a compound of the formula or an easily cleavable ester ox salt thereof with a hetero-aromatic thiolcarboxylic acid of the formula wherein R' is as defined above or a salt thereof to form the desired compound of formula I, or (b) when the starting material includes the desired final substituent in the 3-position of the cephem nucleus: reacting a compound of the formula wherein R' is as defined above or an easily cleavable ester or salt thereof with acid having the formula or an acylating derivative thereof, wherein B is an amino-protecting group and removing said amino-protecting group B
to produce the desired compound of formula I or an easily cleavable ester or pharmaceutically acceptable salt thereof;
and, if desired, in the case of (b) either before or after removal of protecting group B (i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or (ii) converting the product in the form of an easily cleavable ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
(a) when the starting material includes the desired final substituent in the 7-position of the cephem nucleus:
reacting a compound of the formula or an easily cleavable ester ox salt thereof with a hetero-aromatic thiolcarboxylic acid of the formula wherein R' is as defined above or a salt thereof to form the desired compound of formula I, or (b) when the starting material includes the desired final substituent in the 3-position of the cephem nucleus: reacting a compound of the formula wherein R' is as defined above or an easily cleavable ester or salt thereof with acid having the formula or an acylating derivative thereof, wherein B is an amino-protecting group and removing said amino-protecting group B
to produce the desired compound of formula I or an easily cleavable ester or pharmaceutically acceptable salt thereof;
and, if desired, in the case of (b) either before or after removal of protecting group B (i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or (ii) converting the product in the form of an easily cleavable ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
2. A process for preparing a compound of formula I in Claim 1, wherein R and R1 are as defined in Claim 1, or a pharmaceutically acceptable salt thereof, comprising the step of:
reacting a compound of the formula or an easily cleavable ester or salt thereof with a hetero-aromatic thiolcarboxylic acid of the formula wherein R' is as defined above or a salt thereof.
reacting a compound of the formula or an easily cleavable ester or salt thereof with a hetero-aromatic thiolcarboxylic acid of the formula wherein R' is as defined above or a salt thereof.
3. A process for preparing the compound of formula I
in Claim 1 wherein R and R' are as defined in Claim 1, or a pharmaceutically acceptable salt thereof, comprising the step of reacting a compound of the formula wherein R' is as defined above or an easily cleavable ester or salt thereof with acid having the formula or an acylating derivative thereof, wherein B is an amino-protecting group, and removing said amino protecting group B, and, if desired, either before or after said removal of protecting group B (i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or (ii) con-verting the product in the form of an easily cleavable ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
in Claim 1 wherein R and R' are as defined in Claim 1, or a pharmaceutically acceptable salt thereof, comprising the step of reacting a compound of the formula wherein R' is as defined above or an easily cleavable ester or salt thereof with acid having the formula or an acylating derivative thereof, wherein B is an amino-protecting group, and removing said amino protecting group B, and, if desired, either before or after said removal of protecting group B (i) converting the product in the form of the free acid or salt thereof to a corresponding easily cleavable ester or pharmaceutically acceptable salt thereof or (ii) con-verting the product in the form of an easily cleavable ester or salt thereof to the corresponding free acid compound or pharmaceutically acceptable salt thereof.
4. A process according to Claim 2 wherein R is H
and R1 is 2-thiazolyl.
and R1 is 2-thiazolyl.
5. A process according to Claim 3 wherein R is H and R' is 2-thiazolyl.
6. A process according to Claim 2 wherein R is H and R' is 5-methyl-2-thiazolyl.
7. A process according to Claim 3 wherein R is H and R' is 5-methyl-2-thiazolyl.
8. A process according to Claim 2 wherein R is H and R' is 3-methyl-1,2,5-oxadiazol-4-yl.
9. A process according to Claim 3 wherein R is H and R' is 3-methyl-1,2,5-oxadiazol-4-yl.
10. A process as in Claim 4 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
11. A process as in Claim 5 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
12. A process as in Claim 6 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
13. A process as in Claim 7 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
14. A process as in Claim 8 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
15. A process as in Claim 9 followed by the conversion of the product (i) to the corresponding pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester or (ii) to the corresponding sodium or potassium salt.
16. A compound according to formula I in Claim 1, whenever prepared by the process of Claim 1, 2 or 3 or by an obvious chemical equivalent thereof.
17. An acid having the formula or a pharmaceutically acceptable salt thereof, whenever prepared by the process of Claim 4 or 5 or by an obvious chemical equivalent thereof.
18. An acid having the formula or a pharmaceutically acceptable salt thereof, whenever prepared by the process of Claim 6 or 7 or by obvious chemical equivalent thereof.
19. An acid having the formula or a pharmaceutically acceptable salt thereof, whenever prepared by the process of Claim 8 or 9 or by an obvious chemical equivalent thereof.
20. The sodium or potassium salt; or the pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester of the acid:
whenever prepared by the process of Claim 10 or 11 or by an obvious chemical equivalent thereof.
whenever prepared by the process of Claim 10 or 11 or by an obvious chemical equivalent thereof.
21. The sodium or potassium salt; or the pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester of the acid:
whenever prepared by the process of Claim 12 or 13 or by an obvious chemical equivalent thereof.
whenever prepared by the process of Claim 12 or 13 or by an obvious chemical equivalent thereof.
22. The sodium or potassium salt; or the pivaloyloxymethyl, acetoxymethyl, methoxymethyl, acetonyl or phenacyl ester of the acid:
whenever prepared by the process of Claim 14 or 15 or by an obvious chemical equivalent thereof.
whenever prepared by the process of Claim 14 or 15 or by an obvious chemical equivalent thereof.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US05/492,123 US3985740A (en) | 1974-07-26 | 1974-07-26 | 7-[D-(α-amino-α-phenyl-, 2-thienyl- and 3-thienylacetamido)]-3-(5-methylthiazol-2-yl)carbonylthiomethyl-3-cephem-4-carboxylic acids |
| US492636A US3928336A (en) | 1974-07-29 | 1974-07-29 | 7-{8 D-({60 -amino{60 -phenyl-, 2-thienyl- and 3-thienyl-acetamido){9 -3-(3-methyl-1,2,5-oxadiazol-4-yl)carbonylthiomethyl-3-cephem-4-carboxylic acids |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1062701A true CA1062701A (en) | 1979-09-18 |
Family
ID=27050645
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA231,504A Expired CA1062701A (en) | 1974-07-26 | 1975-07-15 | Cephalosporins |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JPS5141380A (en) |
| CA (1) | CA1062701A (en) |
| DE (1) | DE2533376A1 (en) |
| FR (2) | FR2279409A1 (en) |
| GB (1) | GB1492095A (en) |
-
1975
- 1975-07-15 CA CA231,504A patent/CA1062701A/en not_active Expired
- 1975-07-22 FR FR7522802A patent/FR2279409A1/en active Granted
- 1975-07-25 JP JP50090364A patent/JPS5141380A/en active Pending
- 1975-07-25 GB GB31210/75A patent/GB1492095A/en not_active Expired
- 1975-07-25 DE DE19752533376 patent/DE2533376A1/en not_active Ceased
-
1978
- 1978-09-01 FR FR7825300A patent/FR2391729A1/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5141380A (en) | 1976-04-07 |
| FR2279409A1 (en) | 1976-02-20 |
| GB1492095A (en) | 1977-11-16 |
| FR2279409B1 (en) | 1980-05-09 |
| DE2533376A1 (en) | 1976-02-12 |
| FR2391729A1 (en) | 1978-12-22 |
| FR2391729B1 (en) | 1982-08-20 |
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