CA1097612A - Alpha-aminoacyl cephalosporin ethers - Google Patents
Alpha-aminoacyl cephalosporin ethersInfo
- Publication number
- CA1097612A CA1097612A CA186,651A CA186651A CA1097612A CA 1097612 A CA1097612 A CA 1097612A CA 186651 A CA186651 A CA 186651A CA 1097612 A CA1097612 A CA 1097612A
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- Prior art keywords
- cephem
- hydrogen
- carboxylic acid
- amino
- methoxy
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Abstract
ABSTRACT
The present invention relates to a process for preparing .alpha.-aminoacyl cephalosporins represented by the formula
The present invention relates to a process for preparing .alpha.-aminoacyl cephalosporins represented by the formula
Description
This invention relates to a process for preparing the cephalosporin class o~ antibiotics. In particular it relates to a process for preparing a novel group of orally O e~fective cephalosporin antibiotics.
Among the presently used cephat osporin anti~iotics, cephalexin 17-tD-a-phenylglycylamido)~3-methyl-3-cephem-~-carboxylic acid] is a particularly valuable antibiotic because of its oral effectiveness.
In the search for more effective and orally active an~ibiotics a wide variety o~ cephalosporin antibiotics of varying ~tructure~ have been synthesized. Notably, the de~acetoxycephalo~poranic acids (3-methyl-3-cephems~ of which cephalexin i8 a me~ber have been ex~ensively investi-~ gated.
.
.
, ' ~
6~Z
Recently, 7~acylamido-3-methoxymethyl-3-cephem-4-carboxylic acid antibiotics were describe~. The de~cribed compounds have the structural fea~ure of an alkoxy 3ubs~ituted methyl group in the 3~po~i-tion of the dihydro~hiazine ring of the cephalosporin~
The cephalosporin anti~.iotlcs prepa-ed by the process of the present invention repres~nt a novel class of cephalosporin ethers having the oxygen atom of an ther substituent attached directly to the C3 carbon of the dihydrothiazin~ ring~
The cephalo~porin compounds prepared by the process of this invention are represented by the following Formula I, H O H
S ~, R-C-C-N ~
NH2 ~ N ~ o-Rt C\O
wherein R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
ao Rl is hydrogen, methyl, ethyl or 3-methyl-2-but~nyl;
R~ is hydro~en or a ~arboxylic acid protecting ester orming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; wi h the limitation that when Rl is hydrogen, R2 i~ a carboxylic acid protecting ester foxming group.
In the foregoing description of the compounds prepared by the proce~s of the in~ention, ~he t~rm "hydroxy-phenyl," ref~rs to 4~hydroxyphenyl, 3-hydroxyphenyl, and 7~-hydroxyphenyl. "~alophenyl~" refers to ~he i~omeric ~-3g5~ 3 '`''' ' ' ' ~ ' ~7~
fluoro, chloro and bromo phenyl groups such as 4-fluoro-phenyl, 4-chlorophenyl, 3-chlorophenyl, 2-chlorophenyl, 3-bromophenyl, and 4-bromophenyl. "Me~hylphenyl," refers to tho isomeric 2-, 3-, and 4-methylphenyl groups and the texm "me~hoxyphenyl" xefers to 4-methoxyphenyl, 3-methoxy-phenyl and 2-methoxyphenyl.
As used hereirl, the term "a carboxylic acid protect-ing ester forming group," has reference to the ester groups commonly employed for the protection of the C4 carboxyl group of the cephalospo.rins and which are characterized by their ease of removal under basic or acidic hydrolysis conditions or by catalytic hydrogen~lysis to regenerate the free carboxy-lic acid function. Illustrative of such carboxylic acid protectiny groups are the 2,2,2-trichloroethyl, diphenylmethyl (benzhydryl), p-nitrobenzyl, p-methoxybenzyl, t-butyi, and trimethylsilyl. The methods used for the preparation and subsequent removal of the cephalosporin esters described herein are all known methods which have been previously described.
~20 The compounds represented by the Formula I wherein Rl is methyl, ethyl or 3-methyl-2-butenyl and R2 is hydrogen are valuable antibiotics which are effective when administered by the oral route.
The antibiotic compounds are prepared by a variety of ~ynthetic methods. According to one method of prepar~tion a 7-amino-3-hydroxy 3-cephem-4-carboxylic acid es~ex and preferably a salt thereo such as the hydrochloride salt is al~ylated, pre~erably with a diazo compound for example diaæcmethane, to yield the corresponding 3-ether derivative, for example the 3-methoxy or 3-ethoxy derivative. The 7-amino~3-ether cephem ester is then acylated by known methods with a derivative of a substituted ~lycine represented by the formula H O
..
R-C-C-OH
wherein R is as defined for ~ormula I. The carboxylic acid pro-tectin~ ester ~roupr R2, ls removed to provide the antibiotic compound.
Th.e startin~ materials employed in th.e ahove described preparation, 7-amino~3-hydroxy-3-cephem-4-carboxylic acid esters, are prepared, as described in our co-pendin~ Canadian application No. 186,688, filed Nov. 26, 1973, by reacting a 7-acylamido-cephalosporanic acid such as 7-phenoxyacetamidocephalosporanic acid with. a sulfur containing nucleophile accordin~ to known procedures to effect the nucleophilic displacement of the acetox~ group of the cephalosporanic acid and provi.de a 7-acylamido-3-thi:o-substituted methyl-3-cephem-4-carboxylic acid.
. The 3-thi.o-substituted cephem acid is then reduced with zinc/
formic acid in the presence of dimethylformami.de or wi.th. Raney nick.el in the presence of hydrogen to ~ield a 7~acylamido-3-exomethylenecepham-4-carboxylic acid. The acid i5 esteri~i.edr for example ~ith. p-nitrobenzyl bromide and the p-nl.trob.enzyl ester is reacted with phosphorus pentachlor;de in the presence of pyridine to effect the clea~a~e o~ th~e phenoxyacetyl group and provide the 7-amino-3-exomethylenecepham-4-carboxylic acid ester.
The 7~amino-3-exomethylenecepham-4-carboxylic acid ester is then reacted with ozone in an inert solvent at a temperature bet~een about ~80 and 0C. and preferably ; -5-' ' .
$~
between 80 and -50C. to form an intermediate ozonide.
The ozonide i~ decomposed, in situ, in the cold to provide a 7-amino 3-hydroxy-3-cephem-4-carboxylic acid ester as illustrated in the following reaction scheme.
H~N~ O:~ (oZonl(le) r~ OH
C~-O
`0-Rz C\0 0-R~
wherein R2 is a carboxylic acid protecting ester group.
Illustrative o the 7-amino-3-hydroxy~3-cephem-~s-carboxylic acid esters are p-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate, p-methoxybenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate, and 2,2,2-trichloroe~hyl 7 amino-3-hydroxy~3-cephem-4-carboxylate.
According to the process of the present invention, the 7-amino-3-hydroxy-3-cephem-4 carboxylic acid esters are alkylated at the 3-hydroxy group to provide the 3-alkoxy-3-cephem esters. The alkoxylation is carried out by reactin~ the 3~hydroxy ester with a diazo compound, namely ; diazomethane, diazoethane or l-diazo-3-methyl-2~butene.
Al50 ~he 3-hydroxy~3~cephem esters can be alkylated with methyl iodide in the presence of a base, with an alkyl ester of sulfuric acid in the presence of a base, Eor example with dimethyl or diethylsulEate, or with trimethyl oxonium 1uoroborate. Likewise, activated halo compounds in the pre~ence o~ a base can al~o be used to alkylate the 3-hydroxy-3-cephem esters. For example, the a-haloethers such as chloromethyl methyl ether, and bromomethyl ethyl ether;
-haloacid e~ters such as ethyl bromoacetate, ma~hyl chloro-acetate and ~thyl ~-bromopropionate and ~he allylic halides such a~ allyl bromide, allyl chloride and l-bromo-3-methyl-
Among the presently used cephat osporin anti~iotics, cephalexin 17-tD-a-phenylglycylamido)~3-methyl-3-cephem-~-carboxylic acid] is a particularly valuable antibiotic because of its oral effectiveness.
In the search for more effective and orally active an~ibiotics a wide variety o~ cephalosporin antibiotics of varying ~tructure~ have been synthesized. Notably, the de~acetoxycephalo~poranic acids (3-methyl-3-cephems~ of which cephalexin i8 a me~ber have been ex~ensively investi-~ gated.
.
.
, ' ~
6~Z
Recently, 7~acylamido-3-methoxymethyl-3-cephem-4-carboxylic acid antibiotics were describe~. The de~cribed compounds have the structural fea~ure of an alkoxy 3ubs~ituted methyl group in the 3~po~i-tion of the dihydro~hiazine ring of the cephalosporin~
The cephalosporin anti~.iotlcs prepa-ed by the process of the present invention repres~nt a novel class of cephalosporin ethers having the oxygen atom of an ther substituent attached directly to the C3 carbon of the dihydrothiazin~ ring~
The cephalo~porin compounds prepared by the process of this invention are represented by the following Formula I, H O H
S ~, R-C-C-N ~
NH2 ~ N ~ o-Rt C\O
wherein R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
ao Rl is hydrogen, methyl, ethyl or 3-methyl-2-but~nyl;
R~ is hydro~en or a ~arboxylic acid protecting ester orming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; wi h the limitation that when Rl is hydrogen, R2 i~ a carboxylic acid protecting ester foxming group.
In the foregoing description of the compounds prepared by the proce~s of the in~ention, ~he t~rm "hydroxy-phenyl," ref~rs to 4~hydroxyphenyl, 3-hydroxyphenyl, and 7~-hydroxyphenyl. "~alophenyl~" refers to ~he i~omeric ~-3g5~ 3 '`''' ' ' ' ~ ' ~7~
fluoro, chloro and bromo phenyl groups such as 4-fluoro-phenyl, 4-chlorophenyl, 3-chlorophenyl, 2-chlorophenyl, 3-bromophenyl, and 4-bromophenyl. "Me~hylphenyl," refers to tho isomeric 2-, 3-, and 4-methylphenyl groups and the texm "me~hoxyphenyl" xefers to 4-methoxyphenyl, 3-methoxy-phenyl and 2-methoxyphenyl.
As used hereirl, the term "a carboxylic acid protect-ing ester forming group," has reference to the ester groups commonly employed for the protection of the C4 carboxyl group of the cephalospo.rins and which are characterized by their ease of removal under basic or acidic hydrolysis conditions or by catalytic hydrogen~lysis to regenerate the free carboxy-lic acid function. Illustrative of such carboxylic acid protectiny groups are the 2,2,2-trichloroethyl, diphenylmethyl (benzhydryl), p-nitrobenzyl, p-methoxybenzyl, t-butyi, and trimethylsilyl. The methods used for the preparation and subsequent removal of the cephalosporin esters described herein are all known methods which have been previously described.
~20 The compounds represented by the Formula I wherein Rl is methyl, ethyl or 3-methyl-2-butenyl and R2 is hydrogen are valuable antibiotics which are effective when administered by the oral route.
The antibiotic compounds are prepared by a variety of ~ynthetic methods. According to one method of prepar~tion a 7-amino-3-hydroxy 3-cephem-4-carboxylic acid es~ex and preferably a salt thereo such as the hydrochloride salt is al~ylated, pre~erably with a diazo compound for example diaæcmethane, to yield the corresponding 3-ether derivative, for example the 3-methoxy or 3-ethoxy derivative. The 7-amino~3-ether cephem ester is then acylated by known methods with a derivative of a substituted ~lycine represented by the formula H O
..
R-C-C-OH
wherein R is as defined for ~ormula I. The carboxylic acid pro-tectin~ ester ~roupr R2, ls removed to provide the antibiotic compound.
Th.e startin~ materials employed in th.e ahove described preparation, 7-amino~3-hydroxy-3-cephem-4-carboxylic acid esters, are prepared, as described in our co-pendin~ Canadian application No. 186,688, filed Nov. 26, 1973, by reacting a 7-acylamido-cephalosporanic acid such as 7-phenoxyacetamidocephalosporanic acid with. a sulfur containing nucleophile accordin~ to known procedures to effect the nucleophilic displacement of the acetox~ group of the cephalosporanic acid and provi.de a 7-acylamido-3-thi:o-substituted methyl-3-cephem-4-carboxylic acid.
. The 3-thi.o-substituted cephem acid is then reduced with zinc/
formic acid in the presence of dimethylformami.de or wi.th. Raney nick.el in the presence of hydrogen to ~ield a 7~acylamido-3-exomethylenecepham-4-carboxylic acid. The acid i5 esteri~i.edr for example ~ith. p-nitrobenzyl bromide and the p-nl.trob.enzyl ester is reacted with phosphorus pentachlor;de in the presence of pyridine to effect the clea~a~e o~ th~e phenoxyacetyl group and provide the 7-amino-3-exomethylenecepham-4-carboxylic acid ester.
The 7~amino-3-exomethylenecepham-4-carboxylic acid ester is then reacted with ozone in an inert solvent at a temperature bet~een about ~80 and 0C. and preferably ; -5-' ' .
$~
between 80 and -50C. to form an intermediate ozonide.
The ozonide i~ decomposed, in situ, in the cold to provide a 7-amino 3-hydroxy-3-cephem-4-carboxylic acid ester as illustrated in the following reaction scheme.
H~N~ O:~ (oZonl(le) r~ OH
C~-O
`0-Rz C\0 0-R~
wherein R2 is a carboxylic acid protecting ester group.
Illustrative o the 7-amino-3-hydroxy~3-cephem-~s-carboxylic acid esters are p-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate, p-methoxybenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate, and 2,2,2-trichloroe~hyl 7 amino-3-hydroxy~3-cephem-4-carboxylate.
According to the process of the present invention, the 7-amino-3-hydroxy-3-cephem-4 carboxylic acid esters are alkylated at the 3-hydroxy group to provide the 3-alkoxy-3-cephem esters. The alkoxylation is carried out by reactin~ the 3~hydroxy ester with a diazo compound, namely ; diazomethane, diazoethane or l-diazo-3-methyl-2~butene.
Al50 ~he 3-hydroxy~3~cephem esters can be alkylated with methyl iodide in the presence of a base, with an alkyl ester of sulfuric acid in the presence of a base, Eor example with dimethyl or diethylsulEate, or with trimethyl oxonium 1uoroborate. Likewise, activated halo compounds in the pre~ence o~ a base can al~o be used to alkylate the 3-hydroxy-3-cephem esters. For example, the a-haloethers such as chloromethyl methyl ether, and bromomethyl ethyl ether;
-haloacid e~ters such as ethyl bromoacetate, ma~hyl chloro-acetate and ~thyl ~-bromopropionate and ~he allylic halides such a~ allyl bromide, allyl chloride and l-bromo-3-methyl-
2-butene can react with the 3-hydroxy-3-cephem esters to provide respectively the 3-alkoxymethoxy, the 3-carbalkoxy-methoxy and the 3-allyloxy ether derivatives thereof. Th~
preferred alkylatin~ rea~ents are the ~iazo compounds, diazornethane, diazoethane and l-dia20-3-methyl-2-butene.
These diazo compounds react with the 3-hydroxy-3-cephem esters to provide the 3-ethers without side-reaction product ~10 contamination. Other alkylating agents~ for example the activated halo compounds described above, react to provide mixtures of the desired 3-ether with 4~alkylated-3-hydroxy-2-cephem-~-carboxylates and 4-alkylated-3-alkoxy-2-cephem-4-carboxylates. Such mixtures can be separated chromato-graphically to provide the desired 3-ether estar.
The etheriication xeaction is carried out by adding an ~thereal solution of one of the preferred diazo compound~
to a solution of the 3~hydroxy ester in an inert solvent.
The eth~rification proceeds at a sati~factory rate at a temper-ature between about 20 and 25C. An exce~ of the diazo com-pound is desixably employedO
Representative of the 7 amino-3-alkoxy nucleus e~ers which can be prepared and used in this invention are p-nitro-benzyl 7-amino-3-methoxy-3-cephem-4-carboxylate, p-me~hoxy-benzyl 7-ami~o-3-methoxy--3-cephem-4-carboxylate, p-nitrobenzyl 7-amino-3-ethoxy-3~cephem-~-carboxylate, 2,2,2-trichloroethyl 7-amino-3-methoxy-3-cephem-4-carboxylate, p-methoxybenzyl 7-amino-3-~3-methyl-2-but0nyl~1-oxy)-3-cephem~4-carboxyla~e and p-ms~hoxybenzyl 7-amino~3-ethoxy-3-cephem-4-carboxylate.
The 7-amino-3-cephem ~ther acids or esters, prepar~d ' ' ~7~
as de~cribed above, are then acylated with an active deriva-tive of a phenyl or het~rocyclic substituted glycine repre-sented by the formula H O
l 11 R-C-C-OH
N~12 wherein R is as def.in~d for Formula I. The acylation can be caxried out by following the known methods for the acylation of 7-aminocephalosporanic acid and 7-aminodesacetoxycephalo-I sporanic acid. For example, the acid chloride hydrochloride can be reacted with the 7-amino ether nucleus in the presence of a hydrogen halide acceptor~ Hydrogen halide acceptors such as the tertiary amines,pyridine and triethylamine can be used, however the alkylene oxides such as ethylene oxide and propylene oxide are preferred hydrogen halide acceptors in that racemization o the active D-configuration of the D-phenyl, D-thienyl or D-furyl glycyl halide does not occur with these acceptors. Alternatively, the free acid of an amino protected phenyl, thienyl or furyl substituted glycine can be condensed with the amino ether nucleus in the presence of a condensing agent such as a carbodiimide, for example, dicyclohexylcarbodiimide. Also a mixed anhydride ormed with the acylatLng acid and an acid such as formic or acetic acid can be used a~ the acylation reagent in the presence of a con-~ den~ing agent such as N-ethQxycarbonyl-2-ethoxy l,2-dihydro-quinoline (EEDQ).
By other known acylation methods an activated deriva-tive of the phenyl1 thienyl or furylglycines such as an active e~ter, for exampl~, a pentachlorophenyl ester, or an a2ide can he used. In general, any of the known amide coupling .~959 8 ~,~
.~rr~ ' ~ ~ 7 ~ ~ ~
methods can be used in the preparation of the compounds of the invention.
Illustrative o~ the phenyl and h~terocyclic substi-tuted glycines which can be used to acylate ~he 7-amino ether nucleus are the following: D-phenylglycyl chloride hydro-chloride, D-4-hydroxyphenvlglycyl chloride hydrochloride, 2-thienylglycyl chloride hydrochloride, 3-thieny].glycyl chlo-ride hydrochloride, 2-furylglycyl chloride hydrochloride, N-~t-butyloxycarbonyl)-D-phenylglycine, N-(l-carbomethoxy-2-propenyI)-D-phenyl~lycine, and ~- (t-butyloxycarbonyl3 -2-thienylglycin~.
Other amino protecting groups such as the p-methoxy-benæyloxycarbonyl, benzyloxycarbonyl and 2, 2, 2-trichloroethoxy-carbonyl groups can be used to protect the amino group of the : acylating agent. Any of the generally used amino protectin~
groups can be employed, the unction of such groups being merely to protect the reactive amino functio~ during the acylation reaction.
In a spacific emhodiment of ~his invention p-nitro-~0 benzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate hydro-chloride i~ reac ed in methylene chloride with an ethereal solution of diazome~hane to provide p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate~
The 7-amino-3-methoxy-3-cephem estex is then acylated wi~h the mixed anhydride formed with N-(1 carbomethoxy-2-propenyl)-D-phen~lglycine and methyl chloroormate ~n the pxesence of dime~hy1benæylamine a~ about 25C. to provide 7-lN-(l-carbomethoxy-2-propenyl)-D-phenylglycylamido]-3-~ me~hoxy-3-cephem-4-carboxylate. The ~mino protecting group is then removed by acid hydroly~is and the p-nitrobenzyl ester K-3959 g group is removed by catalytic hydrogenolysis with hydrogen in the presence of 5% palladium on carbon catalyst at p~ 2.5 to yield 7-(D-phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid ~Formula I, R = phen~ 1 = methyl and R2 - 11 ) .
The antibiotics can be prepared by an alternative process of the invention hy first acylating ~ 7-amino-3-hydroxy-3-cephem ~ter and then reacting the 7-~phenyl, thienyl or furylglycylamido)-3-hydroxy-3-cephem-4-carboxylic acid product with one of the aforementioned diazo compounds.
The acylation of a 3-hydroxy nucleus ester can be carri~d out by the methods previously described as useful for the acylation of a 7 amino ether nucleus. }lowever, it is preferred to acylate the 3-hydroxy nucleus ester under non-anhydrous acylation methods. When an anhydrous method of acylation is employed N,O-diacylation occurs to some extent to provide a mixture of the desired N acylated product and the N,O-diacylated product~ for example the diacylated product represented by the following generalized formula '~0 0 Il ~ S
R-CH-C-N ~ \ O
NHz ~ L ~ -O~C-CH~R
wher~in R is as defined above and R2 is an ester group. Under non-anhydrous conditions, for example, when the acylation is carried out in w~t acetone, we~ ace~oni~rile or with mixtures of water and wa~er immiscible organic solvPnts, N-acyla~ion occur~ exclusively.
In a further 3peciic embodiment of this invention, p--methoxybenzyl 7-a~ino-3-hydxoxy-3-cephem-4-~arboxylate is acylated with N-(t-butyloxycarbonyl)-D-phenylglycine in the presence of EEDQ to provide p-methoxybenzyl 7-[N-~t-butyloxy-carbonyl~-D-ph~nylglycylamido]-3-hydroxy-3-cephem-4-carboxy-late. The acylated product is xeacted with diazomethane in methylene chloride to provide the ether, p-methoxybenzyl 7-[N-tt-butyloxycarbonyl) D-phenylglycylamido]-3-methoxy-3-cephem-4-carboxylate. The p-metho~ybenzyl group and the t-butyloxycarbonyl amino protectin~ group are removed with trifluoroacetic acid in anisole or p-toluenesulfonic acid in acetonitrile to provide, 7-(D-phenylglycylamido)-3-me~hoxy-
preferred alkylatin~ rea~ents are the ~iazo compounds, diazornethane, diazoethane and l-dia20-3-methyl-2-butene.
These diazo compounds react with the 3-hydroxy-3-cephem esters to provide the 3-ethers without side-reaction product ~10 contamination. Other alkylating agents~ for example the activated halo compounds described above, react to provide mixtures of the desired 3-ether with 4~alkylated-3-hydroxy-2-cephem-~-carboxylates and 4-alkylated-3-alkoxy-2-cephem-4-carboxylates. Such mixtures can be separated chromato-graphically to provide the desired 3-ether estar.
The etheriication xeaction is carried out by adding an ~thereal solution of one of the preferred diazo compound~
to a solution of the 3~hydroxy ester in an inert solvent.
The eth~rification proceeds at a sati~factory rate at a temper-ature between about 20 and 25C. An exce~ of the diazo com-pound is desixably employedO
Representative of the 7 amino-3-alkoxy nucleus e~ers which can be prepared and used in this invention are p-nitro-benzyl 7-amino-3-methoxy-3-cephem-4-carboxylate, p-me~hoxy-benzyl 7-ami~o-3-methoxy--3-cephem-4-carboxylate, p-nitrobenzyl 7-amino-3-ethoxy-3~cephem-~-carboxylate, 2,2,2-trichloroethyl 7-amino-3-methoxy-3-cephem-4-carboxylate, p-methoxybenzyl 7-amino-3-~3-methyl-2-but0nyl~1-oxy)-3-cephem~4-carboxyla~e and p-ms~hoxybenzyl 7-amino~3-ethoxy-3-cephem-4-carboxylate.
The 7-amino-3-cephem ~ther acids or esters, prepar~d ' ' ~7~
as de~cribed above, are then acylated with an active deriva-tive of a phenyl or het~rocyclic substituted glycine repre-sented by the formula H O
l 11 R-C-C-OH
N~12 wherein R is as def.in~d for Formula I. The acylation can be caxried out by following the known methods for the acylation of 7-aminocephalosporanic acid and 7-aminodesacetoxycephalo-I sporanic acid. For example, the acid chloride hydrochloride can be reacted with the 7-amino ether nucleus in the presence of a hydrogen halide acceptor~ Hydrogen halide acceptors such as the tertiary amines,pyridine and triethylamine can be used, however the alkylene oxides such as ethylene oxide and propylene oxide are preferred hydrogen halide acceptors in that racemization o the active D-configuration of the D-phenyl, D-thienyl or D-furyl glycyl halide does not occur with these acceptors. Alternatively, the free acid of an amino protected phenyl, thienyl or furyl substituted glycine can be condensed with the amino ether nucleus in the presence of a condensing agent such as a carbodiimide, for example, dicyclohexylcarbodiimide. Also a mixed anhydride ormed with the acylatLng acid and an acid such as formic or acetic acid can be used a~ the acylation reagent in the presence of a con-~ den~ing agent such as N-ethQxycarbonyl-2-ethoxy l,2-dihydro-quinoline (EEDQ).
By other known acylation methods an activated deriva-tive of the phenyl1 thienyl or furylglycines such as an active e~ter, for exampl~, a pentachlorophenyl ester, or an a2ide can he used. In general, any of the known amide coupling .~959 8 ~,~
.~rr~ ' ~ ~ 7 ~ ~ ~
methods can be used in the preparation of the compounds of the invention.
Illustrative o~ the phenyl and h~terocyclic substi-tuted glycines which can be used to acylate ~he 7-amino ether nucleus are the following: D-phenylglycyl chloride hydro-chloride, D-4-hydroxyphenvlglycyl chloride hydrochloride, 2-thienylglycyl chloride hydrochloride, 3-thieny].glycyl chlo-ride hydrochloride, 2-furylglycyl chloride hydrochloride, N-~t-butyloxycarbonyl)-D-phenylglycine, N-(l-carbomethoxy-2-propenyI)-D-phenyl~lycine, and ~- (t-butyloxycarbonyl3 -2-thienylglycin~.
Other amino protecting groups such as the p-methoxy-benæyloxycarbonyl, benzyloxycarbonyl and 2, 2, 2-trichloroethoxy-carbonyl groups can be used to protect the amino group of the : acylating agent. Any of the generally used amino protectin~
groups can be employed, the unction of such groups being merely to protect the reactive amino functio~ during the acylation reaction.
In a spacific emhodiment of ~his invention p-nitro-~0 benzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate hydro-chloride i~ reac ed in methylene chloride with an ethereal solution of diazome~hane to provide p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate~
The 7-amino-3-methoxy-3-cephem estex is then acylated wi~h the mixed anhydride formed with N-(1 carbomethoxy-2-propenyl)-D-phen~lglycine and methyl chloroormate ~n the pxesence of dime~hy1benæylamine a~ about 25C. to provide 7-lN-(l-carbomethoxy-2-propenyl)-D-phenylglycylamido]-3-~ me~hoxy-3-cephem-4-carboxylate. The ~mino protecting group is then removed by acid hydroly~is and the p-nitrobenzyl ester K-3959 g group is removed by catalytic hydrogenolysis with hydrogen in the presence of 5% palladium on carbon catalyst at p~ 2.5 to yield 7-(D-phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid ~Formula I, R = phen~ 1 = methyl and R2 - 11 ) .
The antibiotics can be prepared by an alternative process of the invention hy first acylating ~ 7-amino-3-hydroxy-3-cephem ~ter and then reacting the 7-~phenyl, thienyl or furylglycylamido)-3-hydroxy-3-cephem-4-carboxylic acid product with one of the aforementioned diazo compounds.
The acylation of a 3-hydroxy nucleus ester can be carri~d out by the methods previously described as useful for the acylation of a 7 amino ether nucleus. }lowever, it is preferred to acylate the 3-hydroxy nucleus ester under non-anhydrous acylation methods. When an anhydrous method of acylation is employed N,O-diacylation occurs to some extent to provide a mixture of the desired N acylated product and the N,O-diacylated product~ for example the diacylated product represented by the following generalized formula '~0 0 Il ~ S
R-CH-C-N ~ \ O
NHz ~ L ~ -O~C-CH~R
wher~in R is as defined above and R2 is an ester group. Under non-anhydrous conditions, for example, when the acylation is carried out in w~t acetone, we~ ace~oni~rile or with mixtures of water and wa~er immiscible organic solvPnts, N-acyla~ion occur~ exclusively.
In a further 3peciic embodiment of this invention, p--methoxybenzyl 7-a~ino-3-hydxoxy-3-cephem-4-~arboxylate is acylated with N-(t-butyloxycarbonyl)-D-phenylglycine in the presence of EEDQ to provide p-methoxybenzyl 7-[N-~t-butyloxy-carbonyl~-D-ph~nylglycylamido]-3-hydroxy-3-cephem-4-carboxy-late. The acylated product is xeacted with diazomethane in methylene chloride to provide the ether, p-methoxybenzyl 7-[N-tt-butyloxycarbonyl) D-phenylglycylamido]-3-methoxy-3-cephem-4-carboxylate. The p-metho~ybenzyl group and the t-butyloxycarbonyl amino protectin~ group are removed with trifluoroacetic acid in anisole or p-toluenesulfonic acid in acetonitrile to provide, 7-(D-phenylglycylamido)-3-me~hoxy-
3-cephem-4-carboxylic acid (Formula I, R = phenyl, R1 = methyl and R2 = H).
Alternatively, the antibiotics can be prepared by employing a 7-~phenyl, thienyl or furylglycylamido)-3-exomethylenecepham-4-carboxylic acid ester as the starting material. According to this method the ~-amino group in the side chain i~ protected with a readily removable amino blocking group for example, the t-butyloxycarbonyl group, the l-carbomethoxy-2-propenyl group, and the 2,2,2-trichloroethoxy-~0 carbonyl group, and the protscted compound is reacted with ozone. The intermedi~te ozonide is decomposed to provide the correspondingLy ~ubstituted 3-hydroxy-3-cephem ester. Alkyl-ation of ~he 3-hydroxy 3-cephem ester and preferably alkyla-tion with a diazo compound, or example diazomethane, yields the 3-methoxy-~ethoxy or 3-methyl-2~butenyl-1 oxy)-3-cephem-
Alternatively, the antibiotics can be prepared by employing a 7-~phenyl, thienyl or furylglycylamido)-3-exomethylenecepham-4-carboxylic acid ester as the starting material. According to this method the ~-amino group in the side chain i~ protected with a readily removable amino blocking group for example, the t-butyloxycarbonyl group, the l-carbomethoxy-2-propenyl group, and the 2,2,2-trichloroethoxy-~0 carbonyl group, and the protscted compound is reacted with ozone. The intermedi~te ozonide is decomposed to provide the correspondingLy ~ubstituted 3-hydroxy-3-cephem ester. Alkyl-ation of ~he 3-hydroxy 3-cephem ester and preferably alkyla-tion with a diazo compound, or example diazomethane, yields the 3-methoxy-~ethoxy or 3-methyl-2~butenyl-1 oxy)-3-cephem-
4-carboxylic acid ester. The carboxylic acid protectinq ester group and the amino protectin~ group are removed ~y known procedures to provide the antibiotic compound of the invention.
Fox example, p-nitrobenzyl 7-[N-(t-butyloxycarbonyl~-D~phenyl-glycylamido]-3-exomethyl~necepham-4-carboxylate is reacted with oæone, the intermediate ozonide is decomposad with sulfur dioxide to provide p-nitr~benzyl 7-~N-~t-butyloxy-carbonyl)-D-phenylylycylamido~-3-hydroxy-3-ceph~m-4-car~oxy-late. The 3-hydroxy e~ter is reacted with diazomethane to provid~ the 3-methoxy derivative. The p-nitrobenzyl ester group is removed from the 3~methoxy derivat$ve by react$ng the ester with hydroyer~ in the presence of 5% palladium on carbon in an acidic medium, ~nd thereafter the t-butyloxy-carbon~l ~roup is removed by acid hydrolysis to provide the anti~iotic 7-[D-phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid.
Following the same procedures the corresponding thienyl and furyl substituted glycyl amides are prepared.
The following compounds are illustrative of the antibiotic compounds of the invention.
7-(D-Phenyl~lycylamido)-3-methoxy-3-cephem~4-carboxylic acid, 7-(D-3-hydroxyphenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7-(D-4-hydroxyphenylgylcylamido)-3-methoxy-3-ceph2m-4-carboxylic acid, 7-(D-2-hydroxyphenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7~D-4-m~hylph2nylglycylamido)-3-msthoxy-3-cephem-4-carboxylic acid, 7-(D-4-chlorophenylglycylamido)-3-methoxy-3-cephem-4 carboxylic acid, 7-(D-3~m2thoxyphenylglycylamido~-3-methoxy-3-cephem-4-carboxylic acid, 7-(D-phenylglycylamido)-3-e~hoxy-3-c~phem-4-car~oxylic acid, X-3~59 12 ~7~
7-~D-phenylglycylamido)~3-(3-me~hyl-2-butenyl-1-oxy)-3-cephem-4-carhoxylic acid, 7-(D-2-th.ienyl~lycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7~(D--2-furyl~lycylamido)-3-methoxy-3-cephem-4--carboxylic acid, 7-(D-3 thienylglycylamido~-3-methoxy-3-cephem-4-carboxylic acid, and 7-~D-2~thienylglycylamido)-3~ethoxy-3-cephem-4-carboxylic ~ acid.
~ preferred group of an~ibiotics prepared by the process Qf this invention are the 3-methoxy-3-cephem acids, Formula I, Rl = methyl and R~ = H .
~mong the preferred 3-methoxy-3-cephem acids the 7-~D-hydroxyphenylglycylamido~-3-methoxy-3-cephe~-4-~arboxylic acid~ ara a further pref~rred group cf antibiotics.
The antibiotics pro~ided by the ~roces~ of this invention lnhibit the growth of microorganis~s pathogenic to man and animals. These antibiotics are a particularly u~2ful cla~s of antibiotics in that they are effective in combating infections caused by gram-positive and gram-negativ~
microorgan~sms whe~ administered parenterally or orally.
~n especially preferred antibiotic compound pr~pared by the process o this invention is 7-(D-phenyl-glycylamido)~3~methoxy-3--cephem-4-carboxylic acid.
In Tabl2 I, which f~llows, the minimum inhibitory concentration (MIC~ for this ~ompound against clinical i~olate3 of penicillin resi~tant S~ lococcus, both in the presence and ab~ence of ~erum, are listed. The MIC values were deter-mined by the Gradient Pl~te technique ~Bryson and Sz~balski, _cience, 116, 45 (1952)._ E-395~ 13 ~7~
able I _ _ -7-~D-Phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid Antibiotic Activity vs. Penicillin Resistant Staphylococcus Clinical I~olate Minimum Inhibitory Concentration (mcg/ml) ~bsent Serum Serum V- ~1 11.4 >20 V-- 32 1~.4 >20 X_~o01 >20 >20 V 84 4.4 11.2 ~- 1.1 0.6 0.5 lethicillin res.istant ~E~h~oco cus.
In Table II below, the MIC values for 7-(D-phenyl-glycylamido)~3-methoxy-3-cephem-4--carboxylic acid against representative gram-ne~ative organisms is presented. The data were obtained by the Gradient Plate technique.
Table II
Antibiotic Activity vs. Gram-Negativ~ Organisms OrganismMinimum Inhibitory Concentration 2a (mcg/ml) ~ ~e~ 7.2 E~cher~ahia coli 6.6 KlebLiel1a pneumoniae 5.1 Aero~act~r a~ 3.6 .
Salmonella ~ 3.8 P~udom~nas ~ >200 Serratia marce~cen~ 124 ___ __ The antimicrobial activity exhibited by 7-(D phenyl-glycyl~mido)-3 methoxy-3-cephem-4-carboxylic acid is illus-~30 tr~tive of the activity of the compounds prepared by th~
~-3~59 14 ~37~
process of the inv~ntion.
A~ previously mentioned the highly preferred csm-pound prepared by the process of this invention, 7-(D-phenyl-glycylamido)-3-methoxy-3-cephem-4-carboxylic acid, is a highly effective anti~iotic when administered orally. For example the ED50 values listed bel~w were obtained with the above named compound in mice infected with the listed micro-organisms.
Microorganism ED50 (mg/kg. oral) _taph~lococcus EX~Lenes 1.0 _ococcus E~e_ o iae 24.1 ~ s a reus 5.2 Escherichia coli 12 2 The ED50 values wexe determined as de~cribed by ~ W. E. Wick et al., Journal of Bacteriology, 81 ~No. 2] 233-; ~35 (1961).
The antibiotic compounds prepared by tha process of this invention can be administered in the free acid form or in ~he form of a pharmaceutically acceptable non toxic salt such as the sodium or pota~sium salt. Such salts are prepared by reacting the antibiotic acid with a sui~abl~
ba~e such a~ sodium carbonate, sodium bicarbonate, sodium hydroxide, or potassium carbonate.
For example, the highl~ preferred antibiotic is eff~ctive $rl combating infections when admini~tered orally at a do~e between about 100 and 500 mg qOi.d. The antibiotic ca~ be administered in a ~ui~able oral pharmaceu ical form for example in gelatin capsule~.
The following examples ar~ provided to further illus-trate the invention.
~-395~ 15 7~
Example 1 p-Nitrobenzyl 7-amino-3-hydroxy-3-cephem 4-carboxylate hydro-chloride.
A solution of 3.85 ~. of p-nitrobenzyl 7-amino-3-methylenecepham-4-car~oxylate hydrochloride in 600 ml. of methanol was cooled i.n an acetone~dry ice bath. Ozone was bubbled through the reaction mixture for approximately 20 minutes at which time the reaction mixture developed a faint blue coloration. Nitrogen was then passed through the reaction mixture to expel excess ozone. Next, the intermediate ozonide was decomposed by passing sulfur dioxide ~as through the reaction mixture until the mixture gave a negative potassium iodide-starch test.
The reaction mixture was evaporated in vacuo and the residue was ~issolved in 200 ml. of 0.lN hydrogen chloride in methylene chloride. The so~ution was evaporated to dryness and the residual reaction product was dissolved in ace-tone.
On cooling, 3.15 ~. o p-nitrobenzyl 7-amino 3-hydroxy-3-cephem-4-carboxylate hydrochloride precipitated as a crystal~
line solid.
I.R. ("Nujol"* Mull): -Carbonyl absorption at
Fox example, p-nitrobenzyl 7-[N-(t-butyloxycarbonyl~-D~phenyl-glycylamido]-3-exomethyl~necepham-4-carboxylate is reacted with oæone, the intermediate ozonide is decomposad with sulfur dioxide to provide p-nitr~benzyl 7-~N-~t-butyloxy-carbonyl)-D-phenylylycylamido~-3-hydroxy-3-ceph~m-4-car~oxy-late. The 3-hydroxy e~ter is reacted with diazomethane to provid~ the 3-methoxy derivative. The p-nitrobenzyl ester group is removed from the 3~methoxy derivat$ve by react$ng the ester with hydroyer~ in the presence of 5% palladium on carbon in an acidic medium, ~nd thereafter the t-butyloxy-carbon~l ~roup is removed by acid hydrolysis to provide the anti~iotic 7-[D-phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid.
Following the same procedures the corresponding thienyl and furyl substituted glycyl amides are prepared.
The following compounds are illustrative of the antibiotic compounds of the invention.
7-(D-Phenyl~lycylamido)-3-methoxy-3-cephem~4-carboxylic acid, 7-(D-3-hydroxyphenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7-(D-4-hydroxyphenylgylcylamido)-3-methoxy-3-ceph2m-4-carboxylic acid, 7-(D-2-hydroxyphenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7~D-4-m~hylph2nylglycylamido)-3-msthoxy-3-cephem-4-carboxylic acid, 7-(D-4-chlorophenylglycylamido)-3-methoxy-3-cephem-4 carboxylic acid, 7-(D-3~m2thoxyphenylglycylamido~-3-methoxy-3-cephem-4-carboxylic acid, 7-(D-phenylglycylamido)-3-e~hoxy-3-c~phem-4-car~oxylic acid, X-3~59 12 ~7~
7-~D-phenylglycylamido)~3-(3-me~hyl-2-butenyl-1-oxy)-3-cephem-4-carhoxylic acid, 7-(D-2-th.ienyl~lycylamido)-3-methoxy-3-cephem-4-carboxylic acid, 7~(D--2-furyl~lycylamido)-3-methoxy-3-cephem-4--carboxylic acid, 7-(D-3 thienylglycylamido~-3-methoxy-3-cephem-4-carboxylic acid, and 7-~D-2~thienylglycylamido)-3~ethoxy-3-cephem-4-carboxylic ~ acid.
~ preferred group of an~ibiotics prepared by the process Qf this invention are the 3-methoxy-3-cephem acids, Formula I, Rl = methyl and R~ = H .
~mong the preferred 3-methoxy-3-cephem acids the 7-~D-hydroxyphenylglycylamido~-3-methoxy-3-cephe~-4-~arboxylic acid~ ara a further pref~rred group cf antibiotics.
The antibiotics pro~ided by the ~roces~ of this invention lnhibit the growth of microorganis~s pathogenic to man and animals. These antibiotics are a particularly u~2ful cla~s of antibiotics in that they are effective in combating infections caused by gram-positive and gram-negativ~
microorgan~sms whe~ administered parenterally or orally.
~n especially preferred antibiotic compound pr~pared by the process o this invention is 7-(D-phenyl-glycylamido)~3~methoxy-3--cephem-4-carboxylic acid.
In Tabl2 I, which f~llows, the minimum inhibitory concentration (MIC~ for this ~ompound against clinical i~olate3 of penicillin resi~tant S~ lococcus, both in the presence and ab~ence of ~erum, are listed. The MIC values were deter-mined by the Gradient Pl~te technique ~Bryson and Sz~balski, _cience, 116, 45 (1952)._ E-395~ 13 ~7~
able I _ _ -7-~D-Phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid Antibiotic Activity vs. Penicillin Resistant Staphylococcus Clinical I~olate Minimum Inhibitory Concentration (mcg/ml) ~bsent Serum Serum V- ~1 11.4 >20 V-- 32 1~.4 >20 X_~o01 >20 >20 V 84 4.4 11.2 ~- 1.1 0.6 0.5 lethicillin res.istant ~E~h~oco cus.
In Table II below, the MIC values for 7-(D-phenyl-glycylamido)~3-methoxy-3-cephem-4--carboxylic acid against representative gram-ne~ative organisms is presented. The data were obtained by the Gradient Plate technique.
Table II
Antibiotic Activity vs. Gram-Negativ~ Organisms OrganismMinimum Inhibitory Concentration 2a (mcg/ml) ~ ~e~ 7.2 E~cher~ahia coli 6.6 KlebLiel1a pneumoniae 5.1 Aero~act~r a~ 3.6 .
Salmonella ~ 3.8 P~udom~nas ~ >200 Serratia marce~cen~ 124 ___ __ The antimicrobial activity exhibited by 7-(D phenyl-glycyl~mido)-3 methoxy-3-cephem-4-carboxylic acid is illus-~30 tr~tive of the activity of the compounds prepared by th~
~-3~59 14 ~37~
process of the inv~ntion.
A~ previously mentioned the highly preferred csm-pound prepared by the process of this invention, 7-(D-phenyl-glycylamido)-3-methoxy-3-cephem-4-carboxylic acid, is a highly effective anti~iotic when administered orally. For example the ED50 values listed bel~w were obtained with the above named compound in mice infected with the listed micro-organisms.
Microorganism ED50 (mg/kg. oral) _taph~lococcus EX~Lenes 1.0 _ococcus E~e_ o iae 24.1 ~ s a reus 5.2 Escherichia coli 12 2 The ED50 values wexe determined as de~cribed by ~ W. E. Wick et al., Journal of Bacteriology, 81 ~No. 2] 233-; ~35 (1961).
The antibiotic compounds prepared by tha process of this invention can be administered in the free acid form or in ~he form of a pharmaceutically acceptable non toxic salt such as the sodium or pota~sium salt. Such salts are prepared by reacting the antibiotic acid with a sui~abl~
ba~e such a~ sodium carbonate, sodium bicarbonate, sodium hydroxide, or potassium carbonate.
For example, the highl~ preferred antibiotic is eff~ctive $rl combating infections when admini~tered orally at a do~e between about 100 and 500 mg qOi.d. The antibiotic ca~ be administered in a ~ui~able oral pharmaceu ical form for example in gelatin capsule~.
The following examples ar~ provided to further illus-trate the invention.
~-395~ 15 7~
Example 1 p-Nitrobenzyl 7-amino-3-hydroxy-3-cephem 4-carboxylate hydro-chloride.
A solution of 3.85 ~. of p-nitrobenzyl 7-amino-3-methylenecepham-4-car~oxylate hydrochloride in 600 ml. of methanol was cooled i.n an acetone~dry ice bath. Ozone was bubbled through the reaction mixture for approximately 20 minutes at which time the reaction mixture developed a faint blue coloration. Nitrogen was then passed through the reaction mixture to expel excess ozone. Next, the intermediate ozonide was decomposed by passing sulfur dioxide ~as through the reaction mixture until the mixture gave a negative potassium iodide-starch test.
The reaction mixture was evaporated in vacuo and the residue was ~issolved in 200 ml. of 0.lN hydrogen chloride in methylene chloride. The so~ution was evaporated to dryness and the residual reaction product was dissolved in ace-tone.
On cooling, 3.15 ~. o p-nitrobenzyl 7-amino 3-hydroxy-3-cephem-4-carboxylate hydrochloride precipitated as a crystal~
line solid.
I.R. ("Nujol"* Mull): -Carbonyl absorption at
5.55 (~-lactam carbonyl) and 5.02 (ester carbonyl hydrogen bonded to 3 hydroxy~ microns.
Electrometric titr tion (66% DMF) pKa 4.0 and 6,3.
Example 2 p-~i~robenzyl 7-amino-3-hydroxy-3~cephem-4-carboxylate~
Four ~illimole o~ p-nitrobenzyl 7-amino-3-hydroxy-3-cepheM-4-carboxylata hydrochloride (prepare~ as described -3~9 * Trademar~ fsr a highly-refinea mineral oil. It is a colorl ss, ~odorless transparent oily liquid comprising a mixture of ~hydrocarbons~
in Example 1) was dissol~ed in water and ethyl acetate was adde~ to the solution. The p~I of the slurry was adjusted from pH 2.2 to pE~ 5 with lN sodium hydroxide. The ethyl acetate layer was separated and was washed with water and dried over ma~nesium sulfate. The dried ethyl acetate layer was evaporated to dryness to yield 1.2 g. of p-nitroben2yl 7-amino-3-hydroxy-3-cephem-4 carboxylate as a crystalline rasidue.
Elemental analysis for: C14EI13N306S:
Theory: C, 47.86; II, 3.73; N, 11.96 Found: C, 47.87; ~I, 4.00; N, 12.11 I.R. (Nujol Mull):
Carbonyl absorption at 5.6S (broad, ~-lactam and ester) and 6.0 (amide) microns.
N.M.R. ~DMS0 d6):
signals at 6.63 (2d, 2H, C2~
5.31 (d, lH, C6H), 4.89 (d, lH, C7~
4.62 (s, 2H, ester CH2), 4.30 (broad s, 2H, 7 N-EI), 2.5-1.8 (m, 4~, aromatic ~i) and 1.2 (d, lH, C30~1) tau.
Example 3 -p-Nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate.
To a stirred suspension of 445 mg. of p-nitro~enzyl 7~amino-3 hydroxy-3-cephem 4-carboxylate hydrochloride (pre-pared as described by ~xample 1) in 35 ml. of dry tetrahydro-furan was added one equivalent of triethylamine followed ~y 10 ml. of an ethereal solution of diazomethane in excess.
A~ter 30 min. the solvent and excess diazomethane were evapo-~-3959 17 .~ . . . . . . . .
~7~ 2 rated and the residue was dissolved in a mixture of water and ethyl acetate. The organic layer was separated and was washed with water and dried. The dried ethyl acetate solution was evaporated to dryness to yield 310 mg. o~ p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate. The product was obtained crystalline by trituration wi~h diethyl ether.
Elemental analysis ~or C15lll$N3O6S:
Theory: C, 49.31; E~, 4.14; N, 11.~0 Found: C, 49.51; H, 4.40; ~1, 11.25 I.R. ("Nujol" Mull): absorption peaks at 2.99 (amide), 5.75 (broad, ~-lactam and ester carbonyl), and 5~98 (amide carbonyl) microns.
U.V. (ethanol~ a~sorption maximum 268 m~, ~-14,~00.
N.M.R. (DMSO d6): signals at 7.10 (broad s, 2H, C7NH2),
Electrometric titr tion (66% DMF) pKa 4.0 and 6,3.
Example 2 p-~i~robenzyl 7-amino-3-hydroxy-3~cephem-4-carboxylate~
Four ~illimole o~ p-nitrobenzyl 7-amino-3-hydroxy-3-cepheM-4-carboxylata hydrochloride (prepare~ as described -3~9 * Trademar~ fsr a highly-refinea mineral oil. It is a colorl ss, ~odorless transparent oily liquid comprising a mixture of ~hydrocarbons~
in Example 1) was dissol~ed in water and ethyl acetate was adde~ to the solution. The p~I of the slurry was adjusted from pH 2.2 to pE~ 5 with lN sodium hydroxide. The ethyl acetate layer was separated and was washed with water and dried over ma~nesium sulfate. The dried ethyl acetate layer was evaporated to dryness to yield 1.2 g. of p-nitroben2yl 7-amino-3-hydroxy-3-cephem-4 carboxylate as a crystalline rasidue.
Elemental analysis for: C14EI13N306S:
Theory: C, 47.86; II, 3.73; N, 11.96 Found: C, 47.87; ~I, 4.00; N, 12.11 I.R. (Nujol Mull):
Carbonyl absorption at 5.6S (broad, ~-lactam and ester) and 6.0 (amide) microns.
N.M.R. ~DMS0 d6):
signals at 6.63 (2d, 2H, C2~
5.31 (d, lH, C6H), 4.89 (d, lH, C7~
4.62 (s, 2H, ester CH2), 4.30 (broad s, 2H, 7 N-EI), 2.5-1.8 (m, 4~, aromatic ~i) and 1.2 (d, lH, C30~1) tau.
Example 3 -p-Nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate.
To a stirred suspension of 445 mg. of p-nitro~enzyl 7~amino-3 hydroxy-3-cephem 4-carboxylate hydrochloride (pre-pared as described by ~xample 1) in 35 ml. of dry tetrahydro-furan was added one equivalent of triethylamine followed ~y 10 ml. of an ethereal solution of diazomethane in excess.
A~ter 30 min. the solvent and excess diazomethane were evapo-~-3959 17 .~ . . . . . . . .
~7~ 2 rated and the residue was dissolved in a mixture of water and ethyl acetate. The organic layer was separated and was washed with water and dried. The dried ethyl acetate solution was evaporated to dryness to yield 310 mg. o~ p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate. The product was obtained crystalline by trituration wi~h diethyl ether.
Elemental analysis ~or C15lll$N3O6S:
Theory: C, 49.31; E~, 4.14; N, 11.~0 Found: C, 49.51; H, 4.40; ~1, 11.25 I.R. ("Nujol" Mull): absorption peaks at 2.99 (amide), 5.75 (broad, ~-lactam and ester carbonyl), and 5~98 (amide carbonyl) microns.
U.V. (ethanol~ a~sorption maximum 268 m~, ~-14,~00.
N.M.R. (DMSO d6): signals at 7.10 (broad s, 2H, C7NH2),
6.22 ~s, 2~1, C2H2)~ 6.20 (5, 3H, C3 methoxyl~, S.27 (d, 1~, C6H), 4.93 (d, 1~, C7H), 4.60 (s, 2H, ester CH2), and 2.35-1.6 (q, 4H, aromatic H) tau.
Ex~ ~le 4 . , O p-Nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate hydrochloride.
To a stirred suspension of 445 mg. of p-nitrobenzyl
Ex~ ~le 4 . , O p-Nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate hydrochloride.
To a stirred suspension of 445 mg. of p-nitrobenzyl
7-amino-3-hydroxy-3-cephem-4-carboxylate hydrochloride in 30 ml. of methylene chloride was added 131 mg. of mono-trimethyl-' silyi acetamide and the mixture was stirred at room tempera-ture for 30 mina An e hereal solution of excess diazomethane -was added and after 20 min. the mixture was evaporated to xemove solvent and excess diazomethane. The residue was trQated with 1 ml. of methanol and then dissolved in an ethyl
8 a~e~ate-water mix~ure. The ethyl ace~ate layer wa~ separated, Sg 18 Ç~
, - ., . '' ~
~7~
washed with water and dried. Hydrogen chloride was passed through the dried ethyl acetate l~yer to precipitate the reaction product, p-nitrobenzy]. 7-amino-3-methoxy-3-cephem-4-carboxylate hydrochloride.
~M.R. (DMSO d6): signals at 6.97 (broad s, 3H, NEI3 ), ' 2 H2)~ 6.23 ¦s, 3E-I, C3 methoxyl), 5 39 (d, l~1, C611), 5~05 (~, lH, C71I) and 2.5-1.92 (q, 4H, aromatic ~I) tau.
Example 5 p-Ni~robenzyl 7-[N-(t-butyloxycarbonyl)-D-phenylglycylamido3-3-methoxy~3-cephem 4-carboxylate.
A mixture o 365 mg. o~ p-nitrobenz~l 7-amino-3-methoxy--3-cephem-4-carboxylate, 256 mg. of N-(t-butyloxycarb-onyl)~-D-~-phenylglycine, and 273 mg. of N-ethoxycarbonyl-2-ethoxy-l,2-dihydroquinoline, in 20 ml. of THF and lO ml. of acetone was stirred at room temperature for 16 hours. The reaction mixture was evaporated to remove volatiles and the residue was dissolv~d in a mixture of water and eth~l ac.etate.
The ethyl acetate layer was separated and was washed with 5 hydrochloric acid and water and was dried. The dried layer was then evaporated to dryness and the product obtained crystalline by ~ri~uration of the residue with diethyl ether.
Elemental analysis or C2~H3~N4OgS
Theory: C, 56.I8; H, 5.05; N, 9.36 Found: C, 55.95; H, 5.l6i N, 9.30 I.R. (Nujol Mull): absorption peaks at 3.01 (amide NH~, 5.67, 5.84, 5.90 and 6.06 Icarbonyl~ microns.
~ N.M.R. (CDCl3~: signals at 8.60 (s, 9H, t-butyl), : 6 75 (~, 2~, C2H~), 6.23 (s, 3H, C3 methQ~yl)/
5.20-3.90 (m, 5H, C6H, sid~-chain CH2, ester CH3 .30 and C7~), 2,80-1.70 (m, 9H, aromatic H) tauO
~-3~59 l9 Example_6 7-~D-Phenylglycylamido)~3-methoxy-3-cep~lem-4-carboxylic acid A aolution of 600 mg. of p-nitrobenzyl 7-[N- (t-~u~tl-oxycarbonyl) D-~-phenylglycylamido3~3-methoxy-3-cephem-4-carboxylate ~prepared as described in Example 5) in 25 ml.
of THF and 6~ ml. o metha~ol containin~ 3 drops of iH hydro-chloric acid was hydrogenated at room temperature under 50 psi hydrogen pressure for 3 hr. in the presence of 600 mg. of pre-reduced 5% palladium-on-carbon.
The catalyst was filtered and was washed with THF.
The filtrate and wash were combined and evaporated to dryness in vacuo. The residue was dissolved in a mixture of water and ethyl acetate and the ~olution was cooled in an ice-water bath~ The pH of the cold solution was adjusted to pH 2.5 and the ethyl acetate layer ~eparated. The ethyl acetate layer was washed, dried and evaporated to dryness to yield 7-EN-(t-butyloxycarbonyl)-D-~-phenylglycylamido]-3-methoxy-3-cephem-4-carboxylic acid as a pale yellow amorphous solid.
The carboxylic acid product was dissolved in 5 ml.
of acetonitrile and 380 mg. of p-toluenesulfonie acid mono-hydrate were added to the solution. The mixture was allowed ~o stand at roo~ temperature for 3 hr. Thereafter 1 ml. of water was added to the mixture and the pH was adjusted to plI
4.5 with triethylamineO
On cooling, 7~(D-~-phenylglycylamido)~3-methoxy-3-cephem-4-carboxylic acid precipitated as a crystalline solid ha~ing ~pec ral propertie~ in agreement with those of the produ~t obtained by Example 8~
~, :
~ ' ' ' ~"-~7~
Exam~e 7 p-Nitrobenzyl 7-[N~ carbomethoxy-2-propenyl)-D-phenylylycyl-amid~-3-methoxy-3-cephem-4-carboxylate~
To 45 ml. of acetonitrile containing 6 drop~ of dimethylbenzylamine were added 815 mg. of methyl 3-a-carboxy-benzylaminocrotonate sodium sa].t and the mixture was cooled in a dxy ice-carbon tetrachloride bath.
To the cold solution were added with stirring 303 mg. of methyl chloroformate and after 20 min. a solution of 1.1 g. of p-nitrobenzyl 7-amino-3-~ethoxy-3-cephem-4-carboxy-late in 45 ml. of acetone was addedO The reaction mixture wa~ stirred in the cold for 30 min. and then at room temper-ature for 2 hr.
The reaction mixture was filtered and evaporated in ; vacuo. The residue was dissolved in ethyl acetate and the solution was washed with water and dried over magnesium sulfate. The dried solution was evaporated to dryness and ; the residue recrystallized from ethanol to yield 1.1 g. of ~ cxy~talline reaction product, p-nitrobenzyl 7-~N-(l-carbo-methoxy-2 propenyl~-D phenylglycylamido]--3~methoxy-3-cephem-4-caxboxylate, melting at about 135 to 145C. with decomposi-: tion.
Elemental analysis or C~8H28N409S:
Theory: C, 5G.37; H, 4.73; N~ 9039 Foun~: C, 56.09; H, 4.57; N, 9027.
7~(D-a-Phenylglycylamido)~3-methoxy-3~cephem-4-carboxylic acid A ~olution of 500 mg. ~f p-nitrobenzyl 7-[N~ carbo-methoxy-2-prop2nyl)-D-phenylglycylamido~-3-me~hoxy-3-~ephem-4-carboxylate (prepared as described by Example 7) in 30 ml.
~-39sg 21 of acetonitrile and 15 ml. of water was acidified to pH 1 with concentrated hydrochloric acid and immediately back titrated to pH 2.5 with lN sodium hydroxide. The mixture was evapo-rated to dryness and th~ residue dlssolved in a solvent mix-ture o~ 40 ml. of TIIF and 80 ml. of methanol.
The ~olution was charged into a Parr low pressure hydrogenation apparatus and hydrogenated under a hydrogen pressure of 50 psi at room temperature for 2.5 hr. in the presence of 500 mg. of 5% palladium-on-carbon. The catalyst ~10 had be~n pre-reduced in ethanol for 30 min. under 50 psi hydrogen pressure at room temperature.
The catalyst was filtered and washed with THF and with water. The filtrate and catalyst wash~s were combin~d and evaporated in vacuo to rem~ve volatile solvents. The aqueou~ residue wa~ slurried with ethyl acetate and the pH
of the slurry was ad~usted to p~I 4.5 with lN sodium hydroxide.
The aqueous layer was separated, washed with ethyl ac~tate and then concentrated in vacuo to a volume of 2 ml.
The aqueous concentrate was diluted with 1 mln of 2~ acetonitrile and the solution was cooled ~o precipi~ate 122 mg. of 7-~D-phenyl~lycylamido)-3-methoxy-3-cephem-4-carboxylic acid as ~he crystallin~ dihydra~e.
EIemen~al analysis for C16H17N305S.2H20:
Theory: C, 48.10; H, 5.30, N, 10.52 Found: C, 47~80; ~, 4.74; N, 10.21.
Nujol Uull): ab~orptio~ peaks a~ 2.95 (amide NH), 5.75, 5.96 t~-lactam ~nd amide carbonyl~), a~d 6.25 ~c~rboxylate) microns.
N.M.R. ~D~0/DCl): signals at 6.58 (2d, 2H~ C~H2~, 6.10 ~, 3H, C3 met~oxyl), 4.87 (d, lH, C6H~, 4.70 (s, X-3959 ~2 lH, ~ -CH), 4.i4 (d, lH, C7H) and 2.41 (s, 5H, aromatic H) tauO
U.~. (pH 7 buffer): ~ max 265 m~u, ~ = 7,500.
Electrometric titration (80% aqueous DMF):
pKa 6.2 and 7.3.
Example 9 Following the procedures described in Examples 7 and 8, p-nitrobenzyi 7-[N~(l-carbomethoxy-2-propenyl)-D-2-thienylglycylamido]-3-methoxy-3~cephem-4-carboxylate is prepared by reacting p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate with methyl 3-[[~2-thienyl3(carboxy)methyl]amino]-crotonate sodi~m salt and methyl chloroformate, and the l-carbomethoxy-2-propenyl amino protecting group is removed by acid hydrolysis followed by the catalytic hydrogenolysis of the p-nitrobenzyl ester group to provide 7-(D-2-thienyl-glycylamido)-3-methoxy-3-cephem-4-carboxylic acid.
Example lO
Following the procedures described in Examples 7 and 8, p-nitrobenzyl 7-~N-(l-carbomethoxy-2-propenyl)-D-4-hydroxyphenylglycylamido~-3-methoxy-3-cephem~4-carboxylate is prepared by reaction of p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate with methyl 3- C~ -carboxy-4-hydroxybenzyl-aminocrotonate sodium .salt and methyl chlorofor~ate. The l-car~ome~hox~-2-propenyl amino protecting group and the .
p-nitrobenzyl ester group are removed to provide 7-(D-4-phenylglycylamido~-3-me~hoxy-3-cephem-4-carboxylic acid.
Example ll p-Nitxokenzyl ~ (D-phenylglycylamido)-3-hydroxy-3-cephem-4-carboxylate.
3~ To a solution of 446 mg. of p-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate in 20 ml. of acetonitrile containing 10 ml. of propylene oxide was added 206 mg. of X-395~ - 23 -D phenylc31ycyl chloride hydrochloride. The re~ction mixture was stirred for 16 hours at room temperature and was then evaporated in vacuo. The residue was tritu~ated with ace-tonitrile to remove soluble impurities. The residue was then ; ~ ' ' ~ ~ I
: i X~3959 - 23a -: ~ : : .
, ; ' , i .
~76~2 dried in vacuo ~o yield 315 mg. of p-nitrobenzyl 7-(D-phenylglycylamido)-3-hydroxy-3-cephem-4-carboxylate.
Elemental analysis for C22H20N~O7S:
Theory: C, 54.54; ~I, 4.16; N, 11.57 Found: C, 54.99; ~1, 4.29; N, 11.02 ~ "
t ~ I.R. (Nujol Mull): absorption peaks at 3.01 (amicle), 5.75 ~-lactam carbonyl), and 6.10 (broad, amide, and hydrogen bonded ester carbonyl) microns.
,. . .
"~
, ~ .
:
X-3g5g 24 , ,,;"
., .
, - ., . '' ~
~7~
washed with water and dried. Hydrogen chloride was passed through the dried ethyl acetate l~yer to precipitate the reaction product, p-nitrobenzy]. 7-amino-3-methoxy-3-cephem-4-carboxylate hydrochloride.
~M.R. (DMSO d6): signals at 6.97 (broad s, 3H, NEI3 ), ' 2 H2)~ 6.23 ¦s, 3E-I, C3 methoxyl), 5 39 (d, l~1, C611), 5~05 (~, lH, C71I) and 2.5-1.92 (q, 4H, aromatic ~I) tau.
Example 5 p-Ni~robenzyl 7-[N-(t-butyloxycarbonyl)-D-phenylglycylamido3-3-methoxy~3-cephem 4-carboxylate.
A mixture o 365 mg. o~ p-nitrobenz~l 7-amino-3-methoxy--3-cephem-4-carboxylate, 256 mg. of N-(t-butyloxycarb-onyl)~-D-~-phenylglycine, and 273 mg. of N-ethoxycarbonyl-2-ethoxy-l,2-dihydroquinoline, in 20 ml. of THF and lO ml. of acetone was stirred at room temperature for 16 hours. The reaction mixture was evaporated to remove volatiles and the residue was dissolv~d in a mixture of water and eth~l ac.etate.
The ethyl acetate layer was separated and was washed with 5 hydrochloric acid and water and was dried. The dried layer was then evaporated to dryness and the product obtained crystalline by ~ri~uration of the residue with diethyl ether.
Elemental analysis or C2~H3~N4OgS
Theory: C, 56.I8; H, 5.05; N, 9.36 Found: C, 55.95; H, 5.l6i N, 9.30 I.R. (Nujol Mull): absorption peaks at 3.01 (amide NH~, 5.67, 5.84, 5.90 and 6.06 Icarbonyl~ microns.
~ N.M.R. (CDCl3~: signals at 8.60 (s, 9H, t-butyl), : 6 75 (~, 2~, C2H~), 6.23 (s, 3H, C3 methQ~yl)/
5.20-3.90 (m, 5H, C6H, sid~-chain CH2, ester CH3 .30 and C7~), 2,80-1.70 (m, 9H, aromatic H) tauO
~-3~59 l9 Example_6 7-~D-Phenylglycylamido)~3-methoxy-3-cep~lem-4-carboxylic acid A aolution of 600 mg. of p-nitrobenzyl 7-[N- (t-~u~tl-oxycarbonyl) D-~-phenylglycylamido3~3-methoxy-3-cephem-4-carboxylate ~prepared as described in Example 5) in 25 ml.
of THF and 6~ ml. o metha~ol containin~ 3 drops of iH hydro-chloric acid was hydrogenated at room temperature under 50 psi hydrogen pressure for 3 hr. in the presence of 600 mg. of pre-reduced 5% palladium-on-carbon.
The catalyst was filtered and was washed with THF.
The filtrate and wash were combined and evaporated to dryness in vacuo. The residue was dissolved in a mixture of water and ethyl acetate and the ~olution was cooled in an ice-water bath~ The pH of the cold solution was adjusted to pH 2.5 and the ethyl acetate layer ~eparated. The ethyl acetate layer was washed, dried and evaporated to dryness to yield 7-EN-(t-butyloxycarbonyl)-D-~-phenylglycylamido]-3-methoxy-3-cephem-4-carboxylic acid as a pale yellow amorphous solid.
The carboxylic acid product was dissolved in 5 ml.
of acetonitrile and 380 mg. of p-toluenesulfonie acid mono-hydrate were added to the solution. The mixture was allowed ~o stand at roo~ temperature for 3 hr. Thereafter 1 ml. of water was added to the mixture and the pH was adjusted to plI
4.5 with triethylamineO
On cooling, 7~(D-~-phenylglycylamido)~3-methoxy-3-cephem-4-carboxylic acid precipitated as a crystalline solid ha~ing ~pec ral propertie~ in agreement with those of the produ~t obtained by Example 8~
~, :
~ ' ' ' ~"-~7~
Exam~e 7 p-Nitrobenzyl 7-[N~ carbomethoxy-2-propenyl)-D-phenylylycyl-amid~-3-methoxy-3-cephem-4-carboxylate~
To 45 ml. of acetonitrile containing 6 drop~ of dimethylbenzylamine were added 815 mg. of methyl 3-a-carboxy-benzylaminocrotonate sodium sa].t and the mixture was cooled in a dxy ice-carbon tetrachloride bath.
To the cold solution were added with stirring 303 mg. of methyl chloroformate and after 20 min. a solution of 1.1 g. of p-nitrobenzyl 7-amino-3-~ethoxy-3-cephem-4-carboxy-late in 45 ml. of acetone was addedO The reaction mixture wa~ stirred in the cold for 30 min. and then at room temper-ature for 2 hr.
The reaction mixture was filtered and evaporated in ; vacuo. The residue was dissolved in ethyl acetate and the solution was washed with water and dried over magnesium sulfate. The dried solution was evaporated to dryness and ; the residue recrystallized from ethanol to yield 1.1 g. of ~ cxy~talline reaction product, p-nitrobenzyl 7-~N-(l-carbo-methoxy-2 propenyl~-D phenylglycylamido]--3~methoxy-3-cephem-4-caxboxylate, melting at about 135 to 145C. with decomposi-: tion.
Elemental analysis or C~8H28N409S:
Theory: C, 5G.37; H, 4.73; N~ 9039 Foun~: C, 56.09; H, 4.57; N, 9027.
7~(D-a-Phenylglycylamido)~3-methoxy-3~cephem-4-carboxylic acid A ~olution of 500 mg. ~f p-nitrobenzyl 7-[N~ carbo-methoxy-2-prop2nyl)-D-phenylglycylamido~-3-me~hoxy-3-~ephem-4-carboxylate (prepared as described by Example 7) in 30 ml.
~-39sg 21 of acetonitrile and 15 ml. of water was acidified to pH 1 with concentrated hydrochloric acid and immediately back titrated to pH 2.5 with lN sodium hydroxide. The mixture was evapo-rated to dryness and th~ residue dlssolved in a solvent mix-ture o~ 40 ml. of TIIF and 80 ml. of methanol.
The ~olution was charged into a Parr low pressure hydrogenation apparatus and hydrogenated under a hydrogen pressure of 50 psi at room temperature for 2.5 hr. in the presence of 500 mg. of 5% palladium-on-carbon. The catalyst ~10 had be~n pre-reduced in ethanol for 30 min. under 50 psi hydrogen pressure at room temperature.
The catalyst was filtered and washed with THF and with water. The filtrate and catalyst wash~s were combin~d and evaporated in vacuo to rem~ve volatile solvents. The aqueou~ residue wa~ slurried with ethyl acetate and the pH
of the slurry was ad~usted to p~I 4.5 with lN sodium hydroxide.
The aqueous layer was separated, washed with ethyl ac~tate and then concentrated in vacuo to a volume of 2 ml.
The aqueous concentrate was diluted with 1 mln of 2~ acetonitrile and the solution was cooled ~o precipi~ate 122 mg. of 7-~D-phenyl~lycylamido)-3-methoxy-3-cephem-4-carboxylic acid as ~he crystallin~ dihydra~e.
EIemen~al analysis for C16H17N305S.2H20:
Theory: C, 48.10; H, 5.30, N, 10.52 Found: C, 47~80; ~, 4.74; N, 10.21.
Nujol Uull): ab~orptio~ peaks a~ 2.95 (amide NH), 5.75, 5.96 t~-lactam ~nd amide carbonyl~), a~d 6.25 ~c~rboxylate) microns.
N.M.R. ~D~0/DCl): signals at 6.58 (2d, 2H~ C~H2~, 6.10 ~, 3H, C3 met~oxyl), 4.87 (d, lH, C6H~, 4.70 (s, X-3959 ~2 lH, ~ -CH), 4.i4 (d, lH, C7H) and 2.41 (s, 5H, aromatic H) tauO
U.~. (pH 7 buffer): ~ max 265 m~u, ~ = 7,500.
Electrometric titration (80% aqueous DMF):
pKa 6.2 and 7.3.
Example 9 Following the procedures described in Examples 7 and 8, p-nitrobenzyi 7-[N~(l-carbomethoxy-2-propenyl)-D-2-thienylglycylamido]-3-methoxy-3~cephem-4-carboxylate is prepared by reacting p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate with methyl 3-[[~2-thienyl3(carboxy)methyl]amino]-crotonate sodi~m salt and methyl chloroformate, and the l-carbomethoxy-2-propenyl amino protecting group is removed by acid hydrolysis followed by the catalytic hydrogenolysis of the p-nitrobenzyl ester group to provide 7-(D-2-thienyl-glycylamido)-3-methoxy-3-cephem-4-carboxylic acid.
Example lO
Following the procedures described in Examples 7 and 8, p-nitrobenzyl 7-~N-(l-carbomethoxy-2-propenyl)-D-4-hydroxyphenylglycylamido~-3-methoxy-3-cephem~4-carboxylate is prepared by reaction of p-nitrobenzyl 7-amino-3-methoxy-3-cephem-4-carboxylate with methyl 3- C~ -carboxy-4-hydroxybenzyl-aminocrotonate sodium .salt and methyl chlorofor~ate. The l-car~ome~hox~-2-propenyl amino protecting group and the .
p-nitrobenzyl ester group are removed to provide 7-(D-4-phenylglycylamido~-3-me~hoxy-3-cephem-4-carboxylic acid.
Example ll p-Nitxokenzyl ~ (D-phenylglycylamido)-3-hydroxy-3-cephem-4-carboxylate.
3~ To a solution of 446 mg. of p-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate in 20 ml. of acetonitrile containing 10 ml. of propylene oxide was added 206 mg. of X-395~ - 23 -D phenylc31ycyl chloride hydrochloride. The re~ction mixture was stirred for 16 hours at room temperature and was then evaporated in vacuo. The residue was tritu~ated with ace-tonitrile to remove soluble impurities. The residue was then ; ~ ' ' ~ ~ I
: i X~3959 - 23a -: ~ : : .
, ; ' , i .
~76~2 dried in vacuo ~o yield 315 mg. of p-nitrobenzyl 7-(D-phenylglycylamido)-3-hydroxy-3-cephem-4-carboxylate.
Elemental analysis for C22H20N~O7S:
Theory: C, 54.54; ~I, 4.16; N, 11.57 Found: C, 54.99; ~1, 4.29; N, 11.02 ~ "
t ~ I.R. (Nujol Mull): absorption peaks at 3.01 (amicle), 5.75 ~-lactam carbonyl), and 6.10 (broad, amide, and hydrogen bonded ester carbonyl) microns.
,. . .
"~
, ~ .
:
X-3g5g 24 , ,,;"
., .
Claims (6)
1. A process for preparing compounds of the formula wherein A is hydrogen or an amino-protecting group;
R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
R1 is hydrogen, methyl, ethyl or 3-methyl-2-butenyl;
R2 is hydrogen or a carboxylic acid protecting ester forming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; with the limitation that when R1 is hydrogen, R2 is a carboxylic acid protecting ester forming group; which comprises reacting a 3-cephem compound of the formula II
wherein R1. and R2 are as defined above, with an acylating reagent of the formula III
or an active derivative thereof wherein R and A are as defined above; optionally reacting the compound thus obtained wherein R1 is hydrogen with diazomethane, diazoethane or 1-diazo-3-methyl-2 butene; and optionally removing the carboxylic acid protecting ester forming group and/or the amino-protecting group.
R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
R1 is hydrogen, methyl, ethyl or 3-methyl-2-butenyl;
R2 is hydrogen or a carboxylic acid protecting ester forming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; with the limitation that when R1 is hydrogen, R2 is a carboxylic acid protecting ester forming group; which comprises reacting a 3-cephem compound of the formula II
wherein R1. and R2 are as defined above, with an acylating reagent of the formula III
or an active derivative thereof wherein R and A are as defined above; optionally reacting the compound thus obtained wherein R1 is hydrogen with diazomethane, diazoethane or 1-diazo-3-methyl-2 butene; and optionally removing the carboxylic acid protecting ester forming group and/or the amino-protecting group.
2. Compounds of the formula I
wherein A is hydrogen or an amino-protecting group;
R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
R1 is hydrogen, methyl, ethyl or 3-methyl-2-butenyl;
R2 is hydrogen or a carboxylic acid protecting ester forming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; with the limitation that when R1 is hydrogen, R2 is a carboxylic acid protecting ester forming group; whenever prepared by the process of claim 1 or by an obvious chemical equivalent thereof.
wherein A is hydrogen or an amino-protecting group;
R is phenyl, hydroxyphenyl, halophenyl, methylphenyl, methoxyphenyl, 2-thienyl, 3-thienyl or 2-furyl;
R1 is hydrogen, methyl, ethyl or 3-methyl-2-butenyl;
R2 is hydrogen or a carboxylic acid protecting ester forming group;
and when R2 is hydrogen the pharmaceutically acceptable non-toxic salts thereof; with the limitation that when R1 is hydrogen, R2 is a carboxylic acid protecting ester forming group; whenever prepared by the process of claim 1 or by an obvious chemical equivalent thereof.
3. A process for preparing 7-(D-phenylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid which comprises a) reacting p-nitrobenzyl 7-amino-3-hydroxy-3-cephem-4-carboxylate hydrochloride with diazo-methane in an inert solvent, and with N-(t-butyloxycarbonyl)-D- .alpha. -phenylglycine, to provide p-nitrobenzyl 7-[N-(t-butyloxycarbonyl)-D- .alpha. -phenylglycylamido]-3-methoxy-3-cephem-4-carboxy-late; and b) removing the p-nitrobenzyl carboxylic acid protecting group and the t-butyloxycarbonyl amino protecting group.
4. 7-(D-phenylglycylamido)-3-methoxy-3-cephem-4--carboxylic acid, whenever prepared by the process of claim 3 or by an obvious chemical equivalent thereof.
5. A process for preparing p-nitrobenzyl 7-(D--phenylglycylamido)-3-hydroxy-3-cephem-4-carboxylate which comprises a) reacting p-nitrobenzyl 7-amino-3-hydroxy-3--cephem-4-carboxylate with D-phenylglycyl chloride hydrochloride.
6. p-Nitrobenzyl 7-(D-phenylglycylamido)-3-hydroxy--3-cephem-4-carboxylate, whenever prepared by the process of claim 5 or by an obvious chemical equivalent thereof.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA186,651A CA1097612A (en) | 1973-11-26 | 1973-11-26 | Alpha-aminoacyl cephalosporin ethers |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA186,651A CA1097612A (en) | 1973-11-26 | 1973-11-26 | Alpha-aminoacyl cephalosporin ethers |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1097612A true CA1097612A (en) | 1981-03-17 |
Family
ID=4098493
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA186,651A Expired CA1097612A (en) | 1973-11-26 | 1973-11-26 | Alpha-aminoacyl cephalosporin ethers |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1097612A (en) |
-
1973
- 1973-11-26 CA CA186,651A patent/CA1097612A/en not_active Expired
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