CA1068260A

CA1068260A - 7-(alpha-furoylureidoaryl and cyclohexadieny-lacetamido) cephalosporin antibiotics

Info

Publication number: CA1068260A
Application number: CA223,505A
Authority: CA
Inventors: Robin D.G. Cooper
Original assignee: Eli Lilly and Co
Current assignee: Eli Lilly and Co
Priority date: 1974-04-01
Filing date: 1975-04-01
Publication date: 1979-12-18
Also published as: GB1502342A; CH614716A5; IL46955A0; AT336790B; BG27235A3; BE827450A; NL7503882A; SU584788A3; AU7963675A; PL95575B1; AR209924A1; FR2265393B1; ZA752005B; FR2265393A1; DE2513954A1; IL46955A; SE7503706L; DD118882A5; JPS50130784A; ES436170A1

Abstract

ABSTRACT OF THE DISCLOSURE

The present invention relates to novel ureido substituted cephalosporin compounds which have a broad antibiotic spectrum against both the gram-positive and gram-negative microorganisms.

Description

1068Z~O

The present invention relates to novel ureido substituted cephalosporin compounds which have a broad ; antibiotic spectrum against both the gram-positive ~ and gram-negative microorganisms.
Several antibiotics of the cephalosporin class have achieved an important status in the treatment and control o~
infectious diseases of man. For example, the well known cephalosporin antibiotic~, cephalothin, cephaloglycin, cephaloridine, and cephalexin have been widely used ln the treatment of infections in man. Considerable e~fort continues to be extended in the development of new cephalosporin anti- `
biotics with increased antibiotic activity and particularly --.
with an expanded spectrum of activity against the gram- -negative microorganisms.
-~ Cephalosporin compounds having a substituted ~
. . .
amino group in the 7-arylacetamido side chain have been pre-viously described. For example, U.S. Patent No. 3,6~6,024 ;i describes certain 7-la-(3-imidoylureido)arylacetamido~-~Z cephalosporanic acids. -Ureido phenylacet~midocephalosporanic acids have been described in U.S. Patent No. 3,673,183.
The compounds provided by the process of th~
inventlon differ structurally from the compounds of the prior ~rt in ~hat the cephalosporin dihydrothiaz~ne rlnq substituted ln the 3-po~ition with a heterocyclic-thiomethyl group. In addition, the cephalosporln antlblotlc-escr~bed h~r~in can be char~cterl~ed ~8 oxp~ndod spoctrum c~ph~losporln ~ntibiotie~ ln that they not only pO8~a-~ tbe : usual high leY~l o~ activity agalnst grDm-posltlve ~lcro-org~ni~ but th~y al~o po8s~ss ~ hlgh lev~l of actl~lty X-~28 ~ ~2-.. _A~.' .

against a broad spectrum of gram-negative microorganisms- which th~i ~rior art compounds did not possess. -`
This invention relates to new cephalosporin anti-biotics represented by the following general formula O O H O H
~ l l 11 1, 1 11 1 S
~z~ -C-N-C-N-CH-C-N ~ ~ :
R' R O ~ ~ CH~-S-R~
COOR
, wherein Z is O; ~
R' is hydrogen or methyl; ; :
R is phenyl, hydroxyphenyl, halophenyl, hydroxy substituted halophenyl, or thienyl;
,,:.

.. . ..
Rl iS
N N
~; I~ 11 .

. R3 or . - - -~;~N - N

S ~ R3 .~
wherein R3 is Cl-C4 lower alkyl; R2 is hydrogen, an indanyl group, a phthal~dyl group,.or an acyloxymeth~l group of the --ormula ~:: O
~ Hz~O-C-Y

i1, ~
.~
!
;X-4283A _3_ ~ .

~ ~ .
s -1068Z~O

wherein Y is Cl-C4 alkyl or phenyl;
and when R2 is hydrogen, the pharmaceutically acceptable salts thereof.
In the above formula I, the term "methylphenyl" refers to the mono and dimethylphenyl groups such as 4-methylpHenyl, ~ 3-methylphenyl, 3,4-dimethylphenyl, or 3,5-dimethylphenyl.
"Hydroxyphenyl" refers to the 3- and 4-monohydroxyphenyl groups, -and to the 3,4-dihydroxy- and 2,4-dihydroxyphenyl groups.
. "Halophenyl" refers to the fluoro, chloro, and bromophenyl 10 groups such as 4-chlorophenyl, 3-chlorophenyl, 3,4-dichloro-phenyl, 4-bromophenyl, or 4-fluorophenyl. "Hydroxy substituted halophenyl" refers to 3-chloro-4-hydroxyphenyl, 3,5-dichloro-4-,, .
~j :i1 ','J,, ':' ~i,; ' ~ .
,~
.~

;~3`
:

:3,~

~ X-4283A -4-hydroxyphenyl, or 3,5-dibromo-4-hydroxyphenyl. "Thienyl" and "furyl" refer to the respective 2- and 3- isomers thereof.
As described above, the heterocyclic radical in the 3-position of the cephem ring is substituted with a Cl-C4 lower `~:
alkyl group. Representative of these groups are the l'methyl-lH-tetrazole-5-yl group, the 1-ethyl-lH-tetrazole-5-yl group, the 5-methyl-1,3,4-thisdiazol-2-yl group, the 5-isopropyl-1,3,4-thiadiazol-5-yl group, and like lower alkyl substituted tetrazole and thiadiazole groups.
The compounds of formula I wherein R' is hydrogen are prepared by reacting a 7-phenylglycylamido, a substituted phenylglycylamido or a 1,4-cyclohexadienylglycylamido 3-tetrazolethiomethyl or thiadiazolethiomethyl substituted . cephalosporin of the following formula II with furoyl or thenoyl isocyanate as illustrated by the following reaction scheme:
' ~' 0 H
.3' 1l 1 ~S
, NH2~CH-C-N~
. R ~N ~CH~--S--Rl II
i ~; 20 OOH

3j ~ ~ 0 :i ~ J C--N=C=0 ''i,~ :: 1 Formula I (R'=R2=H) wherein Rg Rl and Z are as previously defined.
.'' The compounds represented when R' is methyl are .~: prepared by acylating the compound of the formula II with .:~
1~ 30 N-(a-furoyl3-N-methylcarbamoyl chloride or N-(a-thenoyl)-N-. . .
~ X-4283A _5_ . ~

methylcarbamoyl chloride represented by the formula O
C-N=C=0 CH
The carbamoyl chloride is prepared by reacting N-methyl-2-furamide or N-methylthiophene-2-carboxamide with n-butyllithium at -78C. to generate the lithium salt followed by the reaction of the lithium salt with phosgene. The reaction is carried out ~ -in the cold (-78C.) in an inert solvent such as tetrahydro- ~`
furan.
The acylation of the glycylamido cephalosporin of formula II with the carbamoyl chloride is carried out in an inert solvent at a temperature between about -15 and 10C. in the presence of a hydrogen halide acceptor.
Inert solvents such as acetonitrile and tetrahydro-~9 furan can be used conveniently. Hydrogen halide acceptors such as the tertiary amines, triethylamine, and pyridine; and the ~,~ alkylene oxides such as propylene oxide and butylene oxide can s 20 be used. Equimolar amounts of the starting material and the carbamoyl chloride are used. In an example of the preparation i, ~ of a compound of formula I wherein R' is methyl, 7-(D-phenyl-glycylamido)-3~ methyl-lH-tetrazole-5-ylthiomethyl)-3-cephemr4-carboxylic acid is suspended in dry tetrahydrofuran and solubilized by adding bis-(trimethylsilyl)acetamide to the ''9'~ ; suspension- The solution is cooled to about 0C- and an equi-~:
~ molar amount of N-(a-furoyl)-N-methylcarbamoyl chloride in ;'9 9 ~ ~ tetrahydrofuran is added. The mixture is stirred in the cold .~
for about 2 hours, is allo~ed to warm to room temperature, and i .

1068Z6~

the product extracted with an organic solvent such as ethyl acet:ate.
The 7-thienylglycylamido, 7-furylglycylamido, 7-phenylglycylamido and 7-substituted phenylglycylamido-3-hetero-cyclic thiomethyl cephalosporin starting materials of the formula II are prepared by acylating a 7-amino-3-heterocyclic-thiomethyl-substituted cephalosporin nucleus compound with an active derivative of phenylglycine or a substituted phenyl-glycine, for example, the acid chloride, in the presence of - 10 a hydrogen halide acceptor such as triethylamine or sodium carbonate, to provide the acylated phenylglycylamido cephalos-porin starting material.
The compounds of the formula II wherein R is the .
1,4-cyclohexadienyl-1-yl group are prepared by acylation of the 7-amino-3-(1-lower alkyl-lH-tetrazole-5-ylthiomethyl) or (5-lower alkyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid with an active derivative of -amino-a-(1,4-cyclohexadienyl) acetic acid. The cyclohexadienyl acetic acid is converted to an active derivative such as that formed with chloroethyl formate for use as the acylating reagent.
The reaction of the starting material of the formula $~ II with furoyl or thenoyl isocyanate is carried out in the fol-lowing manner. The starting material of the formula II is suspended in an inert solvent at about 20-25C. and a silylat-ing agent such as bis-(trimethylsilyl)acetamide ~BSA) or mono-ilylacetamide (MSA) is added in excess to form a homogenous solution. Inert solvents such as tetrahydrofuran, dichloro-methane, chloroform, or dioxane can be used. After obtaining a solution of the silylated derivative of the starting material, the reaction mixture is cooled in a dry ice-acetone bath to .

.. ~ . . . .

a temperature of approximately -75 to -80C. To the cold solu-tion is added, in excess, the isocyanate. The reaction mixture is then allowed to stir in the cold for about 3 hours and is thereafter allowed to warm to room temperature. Methanol is added to the reaction mixture to decompose excess silylating agent and the mixture is then evaporated under reduced pressure to remove the volatile solvents. The ureido reaction product is then extracted from the residue with ethyl acetate. The product is purified with an acid-base wash and can be further purified by recrystallization.
By way of illustration of the above preparation methods, 7-amino-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid is acylated with phenylglycyl chloride hydrochloride in the presence of sodium carbonaLe to yield 7-phenylglycylamido-3~ methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid. The above acylation product is then reacted with furoylisocyanate following the solubilization of the phenylglycylamido acylation product in tetrahydrofuran with MSA, to provide a compound of the invention wherein R is phenyl, Rl is the l-methyl-lH-tetrazole substituent, R' is H and R2 is hydrogen.
The compounds reprasented by formula I wherein R2 is an acyloxymethyl group are prepared by reacting a salt, for r, example, an alkali metal salt of the free acid compound of formula I with a lower alkanoyloxymethyl halide or with a ;~; halomethyl benzoate. Lower alkanoyloxymethyl halides which can ; be employed are, fsr example, chloromethyl acetate, chloro-methyl propionate, bromomethyl acetate, bromomethyl butyrate, chloromethyl pivaloate, and like halomethyl esters of the lower alkyl straight and branched chain Cl-C4 alkyl carboxylic acidQ.
X-~283A -8-,"

~06826(~

When Y is phenyl, bromo or chloromethylbenzoate can be used in like manner to prepare the benzoyloxymethyl ester. The reaction is carried out by reacting the salt of a cephalosporin acid of the formula I, for example, the sodium or potassium salt with the halomethyl ester in an inert solvent from about 20 to about 55C. Inert solvents which can be employed include, for example, dimethylformamide (DMF), dimethylacetamide (DMAC), tetrahydrofuran, and dioxane. For example, sodium 7-la-(3-furoyl-l-ureido)phenylacetamido]-3-(l-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylate is reacted in aqueous DMF with chloromethyl acetate to provide the acetoxymethyl ester of the cephalosporin acid.
The 5-indanyl esters of the formula I are prepared by esterifying the cephalosporin acid with the phenolic 5-indanol. The indanyl esters can be prepared by the conven-tional procedures used for preparing phenolic esters of carboxylic acids. For example, an active derivative of the cephalosporin acid such as is formed with ethylchloroformate is reacted with 5-indanol.
The phthalidyl esters of the formula I are prepared by reacting bromophthalide with a salt of the cephalosporin acid, for example, the sodium or potassium salt. Bromo- -phth~lide of the formula - Br \~ ~;'`
~, ~ O

is prepared in known manner by the reaction of phthalide with ~, `~ 30 N-bromosuccinimide.
.li `~' ~ X-4283A _g_ . ~ ,,, 'J ~

1068Z6~

The acyloxymethyl esters of the formula I are orally effective forms of the antibiotic acids.
Pharmaceutically acceptable salts of the compounds -represented by formula I are prepared by methods commonly practiced in the cephalosporin art. Representative pharma-ceutically acceptable salts include the alkali metal salts, for example, the sodium, potassium, and lithium salts, the ~ -calcium salt, the ammonium salt, the lower aliphatic ammonium salts, for example, those salts formed with methylamine, dimethylamine, diethylamine, or di-n-propylamine; and the hydroxyalkyl ammonium salts, for example, those formed with ethanolamine or diethanolamine. Preferred pharmaceutically ..,~
acceptable salts include the alkali metal salts, for example, the ~odium salt and the potassium salt. The pharmaceutically acceptable salts of the compounds of formula I are prepared by methods well known in the cephalosporin art. For example, the free acid form of the antibiotic is neutralized with an ,~, .
alkali metal hydroxide or carbonate or with ammonium hydroxide or with the desired alkylamine or ethanolamine to form the salt.
The compounds represented by formula I are il-lustrated by the following compounds.
;;~ 7-la-(3-a-furoyl-1-ureido)-a-phenylacetamido~-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-t3-a-furoyl-1-ureido)-a-(4-hydroxyphenyl)-acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, . ~
7-1-(3-a-furoyl-1-ureido)-a-(a-thienyl)acetamido]-3-(l-ethyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic ~ ~30 acid, `~ X-4283A -10-,: .
~,, 7-[a-(3-a-furoyl-l ureido)-a-(a-furyl)acetamido]-3-(5-isopropyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-~a-(3-a-furoyl-1-ureido)-a-(3-chloro-4-hydroxy-phenyl)acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, : 7-[a-(3-a-furoyl-1-ureido)-a-(3-hydroxyphenyl)-acetamidol-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(1,4-cyclohexadien-l-yl)acetamidol-3-(1-methyl-lH-tetrazole-S-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-la-(3-a-furoY1-1-ureido)-a-(4-chlorophenyl)-acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(4-methylphenyl)- .
'~ acetamido]-3-(5-ethyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-la-(3-a-furoyl-1-ureido)-a-(3,5-dichloro-4-hydroxyphenyl)acetamidol-3-(1-methyl-lH-tetrazole-5-ylthio-methyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(3-bromophenyl)-acetamido~-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, '.il; ~
7-1-(3-a-thenoyl)-1-ureido)-a-phenylacetamido]-3-~1-methyl-lH-tetxazole-50ylthiomethyl)-3-cephem~-4-carboxylic acid, 7-[a-(3-a-thenoyl-1-ureido)-a-(4-hydroxyphenyl)-~i~ acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-~; carboxylic acid, .~ ~

:
":

7-[a-(3-a-thenoyl-1-ureido)-a-(-thienyl)acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-thenoyl-3-methyl-1-ureido)-~-phenyl-acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-~-thenoyl-1-ureido)-a-phenylacetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-3-methyl-1-ureido)--phenyl-' acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, -7-[a-(3-a-furoyl-3-methyl-1-ureido)-a-~a-thienyl)-acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3- -cephem-4-carboxylic acid, ; 7-[a-(3-a-furoyl-3-methyl-1-ureido)-a-~3-chloro-4-- hydroxyphenyl)acetamido]-3-(1-methyl-lH-tetrazole-5-ylthio-;~, .
methyl)-3-cephem-4-carboxylic acid, ~i 7-la-(3-a-furoyl-3-methyl-1-ureido)-a-~4-hydroxy-~;~ 20 phenyl)acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthio-methyl)-3-cephem-4-carboxylic acid, and the pharmaceutically . . .
acceptable non-toxic salts thereof.
The compounds represented by formula I and the pharmaceutically acceptable salts thereof inhibit the growth of microorganisms ~athogenic to animals and man. In particular, these compounds inhibit the growth of a broad spectrum of gram-negative and gram-positive microorganisms. They are further aative in inhibiting the growth of penicillin resis-tant Staphylococcus organisms. Accordingly, the compounds of ~30 the invention are useful in combating infections in animals and ~'~ X-4293A -12-.,~ .

~0682t;0 man attributable to gram-positive and gram-negative micro-organisms. The furoyl and thenoylureido-cephalosporins are effective against gram-negative microorganisms of the indole-positive and indole-negative Proteus sp., the Aerobacter sp., the Pseudomonas, the Enterobacter sp., the Serratia, e.g., S. marcescens, Escherichia coli, and Klebsiella. They are also highly effective against the Streptococcus D group of bacteria as well as Staphylococcus aureus and penicillin resistant strains of Staphylococcus. ~
The compounds of formula I can be administered -by the parenteral route, for example, intramuscularly or intravenously. When administered in non-toxic doses ranging between about 25 and about 1,000 mg. per kg. of the patient's body weight, the compounds are effective in the treatment of infectious diseases attributable to both the gram-positive and gram-negative microorganisms. The compounds of this invention can be formulated for such administrative routes as aqueous suspensions or solutions suitable for injection.
For example, the compounds of the invention, as the alkali metal salts ! can be employed in sterile aqueous solutions for injection or they can be prepared as sterile suspensions in an inert pharmaceutical carrier suitable for injection.
When administered intravenously, the salt form of the compound of the invention, for example, the sodium salt, can be dis-solved in one of the standard clinical I.V. ~:olutions, for example, I.V. dextrose, for administration via I.V. drip.
Preferred compounds are those represented by formula I wherein R is phenyl, hydroxyphenyl, halophenyl, hydroxy substituted halophenyl, or thieny:L.

.

1068;~60 An especially preferred group of compounds are those represented when R is phenyl, hydroxyphenyl, or hydroxy sub-stituted halophenyl especially hydroxy substituted chlorophenyl, and the pharmaceutically acceptable non-toxic salts thereof.
The preferred compounds described above are illustrated by:
7-[~-(3-a-furoyl-1-ureido)-a-phenylacetamido]-3-(l-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[~-(3-a-furoyl-1-ureido)-a~(4-hydroxyphenyl)-acetamido]-3-(5-methyl-1,3,4-thiadiazole-2~ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(3-hydroxyphenyl)-acetamido]-3~ methyl-lH-tetrazole-5-ylthiomethyl)-3-ceph0m-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(3-chloro-4-hydroxy-phenyl)acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, 7-[a-(3-a-furoyl-1-ureido)-a-(3,5-dichloro-4-hydroxyphenyl)acetamido]-3-(5-methyl-1,3,4-thiadiazole-2-, 20 ylthiomethyl)-3-cephem-4-carboxylic acid, ,` 7-[a-(3-a-furoyl-3-methyl-1-ureido)--(4-hydroxy-phenyl)acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid, and the pharmaceutically a~ceptable non-toxic salts thereof.
'~ The antibiotic activity of the compounds of formula I
is illustrated by the in vitro data presented in the following Tables I and II for two of the preferred compounds. In the tables, the minimum inhibitory concentrations ~MIC) of the test compounds versus the indicated gram-positive and gram-negative microorganisms is presented. The MIC values were determined by .

~0682~0 the gradient plate method which is essentially the methoddescribed by Bryson and Szybalski, Science, 116, 45-46 (1952).
Table I lists the in vitro antibiotic activity ; demonstrated by the test compounds against representative gram-negative microorganisms. Table II lists the inhibitory activity in terms of MIC values against clinical isolates of - penicillin resistant Staphylococcus microorganisms both in the presence of and in the absence of serum.
TABLE I
Antibiotic Activity of 7-[a-(3-Furoyl-l-ureido)phenyl-(hydroxyphenyl)acetamido]cephalosporins vs. Gram Microorganisms MIC (mcg/ml) Test Compound Test Organism Al B2 Shiqella sp. 5.5 5.5 Escherichia coli 7.0 5.8 Kleb~iella pneumoniae5.0 6.3 Aerobacter aeroqenes 7.5 6.5 Salmonella _eidelberg6.8 5.8 Pseudomona~ aeru~inosa 12.3 10.7 Serratia marceCicens19.5 14.5 .
".9 . Test Compound A=7-la-~3-a-furoyl-1-ureido)-a-phenylacet-amido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid.
~,~ 2 Test Compound B=7-[a-(3-a-furoyl-1-ureido)-a-(4-hydroxy-i phenyl~acetamido]-3 (1-methyl-lH-tetrazole-5-ylthiomethyl)-,~ 3-cephem-4-carboxylic acid.
,,.~

, '''. .

., > X-4283A ~15-TABLE II
Antibiotic Activity of 7-la-(3-Furoyl-l-ureido)-(hydroxyphenyl)acetamidocephalosporins vs.
Resistant Staphylococcus Resistant MIC (mcg/ml) Staph. Test Compound -A B

V-41 3.0 8.0 5.0 ~20 V-32 4.5 8.0 13.7 >~0 -X-400 >20 ~20 >20 >20 ` V-84 0.4 1.0 1.0 1.0 Xl.l 0.4 1.0 1.0 1.0 ~1 1 Test compounds A & B are respectively the test compounds ,, of Table I.
Compound tested in the absence of serum.
Compound tested in the presence of serum.

As indicated by the in vitro data presented above 3 for two of the preferred compounds, the furoylureido cephalos-~ porin compounds disclosed herein are resistant to the action Ij of the enzymes, penlcillinase and cephalosporinaqe, generated ~!~; respectively by the penicillin-resistant Staphylococci and the gram-negative organisms.
The preparation of the compounds represented by formula I is further illustrated by the following examples.
Example 1 To a suspension of 0.6693 g. of 7~phenylglycyl-amido-3~ methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid in 100 ml~ of dry tetrahydrofuran was added an excess of monosilyl acetamide to form a solution. Linde 4A molecular si~ve was added and the mixture was cooled in , .

1~682f~0 - dry ice-ace~one bath. An excess of furoyl isocyanate was added to the cold mixture with stirring. Stirring was con-tinued in the cold for 2 hours and then the mixture was al-lowed to warm to room temperature. Fifty milliliters of methanol were added and the reaction mixture was filtered.
The filtrate was evaporated under reduced pressure to remove volatile solvents. The residue was dissolved in aqueous sodium bicarbonate and the solution was extracted with ethyl acetate. The aqueous phase was acidified to about pH 1.5-

2.0 with dilute hydrochloric acid and was extracted with ethyl acetate. The extract was concentrated and was then diluted with about an equal volume of petroleum ether to pre-cipitate the product, 7-[a-(3-a-furoyl-1-ureido)-a-phenyl-acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid.
The nuclear magnetic resonance spectrum of the product run in deuterated dimethyl sulfoxide was in agree-ment showing peaks at ~5.10 and ~5.10 for the C6 and C7 ~-', lactam protons; multiplets at ~4.32 and ~3.65 for the methylene , 20 protons and a singlet at ~4.00 for the N-methyl protons of , ~ the tetrazole group.
;Y~ Example 2 To a suspension of 0.955 g. of 7-(4-hydroxyphenyl-glycylamido)-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-.
cephem-4-carboxylic acid in 100 ml. of tetrahydrofuran was ~ added excess monosilylacetamide. To the resultant solution i~t~ was added molecular sieve (Linde 4A molecular sieve) and the mixture cooled in a dry-ice acetone bath. A slight excess of furoyl isocyanate was then added with stirring.
`; 30 The reaction mixture was stirred in the cold for 3 hours ~ X-4283A -17-... .
i : .

'--and~was then allowed to warm to room temperature. Methanol, 100 ml., was added and ~he mixture was then evaporated under reduced pressure to remove volatile solvents. The residue was dissolved in an aqueous solution of sodium bicarbonate and the solution was washed with ethyl acetate. The solution was then acidified with dilute hydrochloric acid to about pH 2 and extracted with ethyl acetate. The extract was concentrated and the concentrate diluted with petroleum ether to precipitate the reaction product, 7-[a-(3-furoyl-1-ureido)-4-hydroxyphenylacetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl~-3-cephem-4-carboxylic acid. The product was further purified by crystallization from methanol-diethyl ether-pentane ~nSkelly-solve-Bn*).
Example 3 Following the procedures of Example 1, furoyl isocyanate is reacteb with 7-phenylglycylamido-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-carboxylic ~cid to obtain 7-la-(3-a-furoyl-1-ureido)--phenylacetamido]-3-(5-methyl-1,3,4-thiadiazole-2-ylthiomethyl)-3-cephem-4-, .
, 20 carboxylic acid.
.~; .
Example 4 ~_ 7-[a-(3- a-Furoyl-l-ureido)-~-(3-chloro-4-hydroxy-phenyl)aeetamido]-3-~1-methyl-lH-tetrazole-5-ylthiomethyl)-3-u~ cephemr4-~arboxylic acid is prepared by the reaction of .
.
furoyl i~ocyanate with 7-~3-c~loro-4-hydroxyphenyl~lycyl-a~ido)-3-~1-methyl-lH-tetra~ole-5-ylthiomethyl)-3-ceph~mr~-.~
;, ~ carboxylic acid.
.~
Example 5 To a ~uspen~ion of ~61 ~g 7-phenylglycyl~m~do-3-.' 1 30 (l-methy~ t~tra~ol~-s-ylthio~ethyl)-3-c~p~Qmr4-c~rboxyl~c .
~i X-~83A -18-, .
*Trademark t acid in 8 ml. of dry acetonitrile containing 2 ml~ of propylene oxide was added with stirring 1 ml. of bis-(tri-methylsilyl)acetamide (BSA). The resulting orange solution was cooled to 0C. and a solution of a slight molar excess of N-(2-furoyl)-N-methylcarbamoyl chloride in 2 ml. of dry acetonitrile was added. The reaction mixture was stirred for 2 hours in the cold and was then allowed to warm to room temperature.
The reaction mixture was filtered and the methanol was added to the filtrate to destroy any excess BSA present.
The filtrate was evaporated and the residue was dissolved in a mixture of ethyl acetate and water; The pH of the mixture was adjusted to 2 and the organic layer separated. The organic layer was washed with water, dried, and evaporated to yield the reaction product, 7-~a-~3-a-furoyl-3-methyl-1-ureido)-a-phenylacetamido]-3-(1-methyl-lH-tetrazole-5-ylthio-, methyl)-3-cephem-4-carboxylic acid. The product was crystal-lized from acetone-ether to yield 156 mg. of purified product.
; Example 6 ` To a solution of 511 mg. of 7-(3-chloro-4-hydroxy-phenylglycylamido)-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-

3-cephem-4-carboxylic acid in 1 ml. of tetrahydrofuran and $~
1 ml. of bis-(trimethylsilyl)acetamide was added a solution o~ 2 ml. of propylene oxide in 8 ml. of acetonitrile. The -resulting solution was cooled to 0C. and a solution of 2 ml. of N-(2-furoyl)-N-methylcarbamoyl chloride in 2 ml.
i~ of acetonitrile was added. The dark reaction mixture was stirred at 0C. for 2 hours and was then allowed to warm to room temperature. The mixture was filtered and 3 ml. of ' lO methanol were added to the filtrate to destroy any excess ~,X-4283A -19-.~

106~Z60 silyl reagent. The filtrate was evaporated and the residue dissolved in a mixture of ethyl acetate and water. With -stirring the pH of the mixture was adjusted to 2 with dilute hydrochloric acid. The organic phase was separated and was wa~hed with water, dried, and evaporated under reduced pressure. The reaction product residue, 7-[a-(3-a-furoyl-3-methyl-l-ureido)-a-(3-chloro-4-hydroxyphenyl)acetamido]-3-(l-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid was crystallized from a mixture of ethyl acetate-diethyl ether-petroleum ether to yield 74 mg. of crystalline product.
Example 7 7-la-~3--furoyl-1-ureido)-a-(a-thienyl)acetamido]-3-(l-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid.
To a su~pen~lon of 467 mg of 7-la-(a-thienyl)-a-~; amino-acetamido~-3-(1-methyl-lH-tetrazole-5-ylthlomethyl)-3-~;~ cephem-4-carboxylic acid in 100 ml of dry THF were added 2 g of mono-(trimethylsilyl)acetamlde (MSA). When solution had ^~ occurred a small amount of molecular sieve was added to maintain dryne~s and the solution was cooled to 0C. To the cold solution was added a solution of an excess of furoyl isocyanate in 2 ml of THF. The reaction mixture was stirred at 0C. for 3 hours and was then allowed to warm to room ; temperature. The reaction mixture was filtered and 5 ml ; of methanol were added to the filtrate. The filtrate was evaporated and the re~idue was layerad with ethyl acetate and water. The pH of the aqueous phase was adjusted to 2 with dilute hydrochloric acid and the organic layer was separated. The organic layer was treated with a dilute solution of sodium bicarbonate to pH 7.2. The aqueous layer . . .
'~,~. , ~ ,............ . .. . .. ..

~- ;
iO68~60 was separated and acidified to pH 2 with dilute hydrochloric acid at ice bath temperature. The acidified solution was extracted with ethyl acetate. The extract was dried and evaporated and the residue recrystallized from a mixture of acetone-diethyl ether-petroleum ether to yield a first crop of product weighing 45 mg, a second crop weigning 83 mg and an additional 24 mg of product from the filtrate.
Example 8 7-[a-(3-a-furoyl-3-methyl-1-ureido)-a-(1,4-cyclohexadienyl)-acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-

4-carboxylic acid.
By following the procedures described in Example 5, 463 mg of impure 7-la-(1,4-cyclohexadienyl)-a-amino-acetamido]-3-~1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic ', acid was dissolved in 15 ml of methylene chloride containing excess bis-(trimethylsilyl)acetamido (~SA), propylene oxide x was added to the solution followed by a slight molar excess of N-(2-furoyl)-N-methylcarbamoyl chloride. After stirring for 2 hours in the cold 74 mg of the product was recovered ;~ 20 as an amorphous powder.
~, .
.i Example 9 .~, 7-[a-(3-~-thenoyl-1-ureido)-~-(-thienyl)acetamido]-3-(l-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid.
To a solution of 467 mg of 7-(a-amino-a-thienyl-acetamido)-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid in 100 ml of dry THF, formed by ,1:
the addition of 1 ml of BSA, was added at 0C. a solution containing a molar excess of a-thienylisocyanate in 2 ml of dry THF. The reaction mixture was stirred for 3 hours .i , ~0682~0 at 0C. and was then allowed to warm to room temperature.
The product was recovered from the reaction mixture by fol]owing the isolation procedures described in Example 7.
The product was recrystallized from a mixture of acetone-diethyl ether-petroleum ether to yield 156 mg.
Example 10 7-[~-(3--furoyl-3-methyl-1-ureido)-a-(2-thienyl)acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid.
A suspension of 234 mg of 7-l(a-amino-a-(a-thienyl)-acetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid in 50 ml of dry acetonitrile was solubilized -~
with 5 ml of BSA and the slightly orange solution was cooled -to 0C. To the cold solution were added 2 ml. of propylene oxide and a stoichiometric excess of N-(a-furoyl)-N-methyl-carbamoyl chloride. The reaction mixture was stirred 2 hours at 0C-, one hour at room temperature and methanol was added to decompose the silylating agent. The mixture was evaporated and the residue extracted with ethyl acetate at pH2. The . . 20 extract was washed with water, dried and evaporated under vacuum. The dried residue was recrystallized from a mixture of acetone-diethylether-petroleum ether to yield 114 mg of the product.

i:~
~ ~ .

X-4283~ -22-. ~ .

Claims

The embodiments of the invention for which an ex-clusive property or privilege is claimed are as follows:

1. A process for preparing ureido substituted cephalosporin compounds of the formula I
wherein z is O;
R' is hydrogen or methyl;
R is phenyl, hydroxyphenyl, halophenyl, hydroxy substituted halophenyl, or thienyl;

R1 is or wherein R3 is C1-C4 alkyl;
R2 is hydrogen, indanyl, phthalidyl, or an acyloxy-methyl group of the formula wherein Y is C1-C4 alkyl or phenyl;
and when R2 is hydrogen, the pharmaceutically acceptable non-toxic salts thereof;
which comprises reacting a 7-glycylamidocephalosporin compound of the formula II

with a compound of the formula III
wherein A is -N=C=O or ;

and if desired converting the acid so obtained wherein R2 is hydrogen to the corresponding ester wherein R2 is other than hydrogen; and where desired, when R2 is hydrogen, forming the pharmaceutically acceptable non-toxic salts of said compound of formula I.

2. Compounds of the formula I wherein Z, R, R', R1, and R2 are as defined in claim 1, when prepared by the process of claim 1 or by an obvious chemical equivalent thereof.

3. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-1-ureido)-.alpha.-phenylacetamido]-3-(1-methyl-1H-tetrazole-5-ylthio-methyl)-3-cephem-4-carboxylic acid which comprises reacting 7-phenylglycylamido-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with furoyl isocyanate.

4. 7-[.alpha.-(3-.alpha.-Furoyl-1-ureido)-a-phenylacetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 3 or by an obvious chemical equivalent thereof.

5. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-1-ureido)-.alpha.-(4-hydroxyphenyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-(4-hydroxyphenyl)glycylamido-3-(1-methyl-1H-tetra-zole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with furoyl isocyanate.

6. 7-[.alpha.-(3-.alpha.-Furoyl-1-ureido)-.alpha.-(4-hydroxyphenyl)-acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 5 or by an obvious chemical equivalent thereof.

7. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-1-ureido)-.alpha.-(3-chloro-4-hydroxyphenyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-(3-chloro-4-hydroxyphenyl)glycylamido-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with furoyl isocyanate.

8. 7-[.alpha.-(3-.alpha.-Furoyl-1-ureido)-.alpha.-(3-chloro-4-hydroxy-phenyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl-3-cephem-4-carboxylic acid when prepared by the process of claim 7 or by an obvious chemical equivalent thereof.

9. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-3-methyl-1-ureido)-.alpha.-(3-chloro-4-hydroxyphenyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-(3-chloro-4-hydroxyphenyl)-glycylamido-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with N-(2-furoyl)-N-methylcarbamoyl chloride.

10. 7-[.alpha.-(3-.alpha.-furoyl-3-methyl-1-ureido)-a-(3-chloro-4-hydroxyphenyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthio-methyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 9 or by an obvious chemical equivalent thereof.

11. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-3-methyl-1-ureido)-.alpha.-phenylacetamido]-3-(1-methyl-lH-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-phenylglycylamido-3-(1-methyl-1H-tetrazole-5-ylthio-methyl)-3-cephem-4-carboxylic acid with N-(2-furoyl)-N-methyl-carbamoyl chloride.

12. 7-[.alpha.-(3-.alpha.-Furoyl-3-methyl-1-ureido)-.alpha.-phenyl-acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 11 or by an obvious chemical equivalent thereof.

13. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-1-ureido)-.alpha.-(.alpha.-thienyl)acetamidol-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-[.alpha.-amino-.alpha.-(2-thienyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with furoyl isocyanate.

14. 7-[.alpha.-(3-.alpha.-Furoyl-1-ureido)-.alpha.-(.alpha.-thienyl)-acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 13 or by an obvious chemical equivalent thereof.

15. The process for preparing 7-[.alpha.-(3-.alpha.-furoyl-3-methyl-1-ureido)-.alpha.-(.alpha.-thienyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid which comprises reacting 7-[.alpha.-amino-.alpha.-(2-thienyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid with N-(2-furoyl)-N-methylcarbamoyl chloride.

16. 7-[.alpha.-(3-.alpha.-Furoyl-3-methyl-1-ureido)-.alpha.-(.alpha.-thienyl)acetamido]-3-(1-methyl-1H-tetrazole-5-ylthiomethyl)-3-cephem-4-carboxylic acid when prepared by the process of claim 15 or by an obvious chemical equivalent thereof.