AU463529B2 - Cephalosporin derivatives and process fob the preparation thereof - Google Patents

Description

37,950 P2 This invention relates to new synthetic compounds

of value as antibacterial agents, as nutritional supple-

ments in animal feed, as agents for the treatment of mastitis

in cattle, and as therapeutic agents in poultry and animals,

including man, in the treatment, particularly by oral adminis-

tration, of infectious diseases caused by many Gram-positive and

Gram-negative bacteria.

More particularly, the invention relates to certain

7-[D-(a-amino-a-phenyl-,2-thienyl- and 3-thienyl-acetamido)]- 4 3-rS-(1,2,3-triazole-5-yl)thiomethyl]-3-cephm- carboxylic

acids,.their nontoxic, pharmaceutically acceptable salts and

to the process for the preparation thereof.

Cephalothin and cephaloridine are well-known antibac-

terial agents; see U.S. patents 3,218,318; 3,449,338 and

3,498,979. The literature also contains considerable data on the activity of cephaloglycin and cephalexin; see U.S. patents

3,303,193 and 3,507,861 and Great Britain 985,747 and 1,054,806. Newer cephalosporins include cefazolin and cephapirin; see

U.S. patent 3,516,997 [and also Netherlands 68/05179 (Farmdoc

34,328) and South Africa 68/4513] and U.S. patent 3,422,100.

The literature on cephalosporins has been reviewed by E.P. Abraham, Quart. Rev. (London) 21, 231 (1967), by E. Van

Heyningen, Advan. Drug Res., 4, 1-70, (1967) and briefly in

Annual Reports in Medicinal Chemistry, Academic Press, Inc.,.

III Fifth Avenue, New York, New York, 10003, by L.C. Cheney on pages 96 and 97 (1967) and by K. Gerzon and R.B. Morin on

pages 90-93 (1968). New cephalosporins are frequently reported at

4635.29

37,950- 1n Agents on Antimicrobial the annual Interscience Conference by Sassiver et al., Anti- and Chemotherapy as illustrated 1968, American Society microbial Agents and Chemotherapy 101-114 (1969) for Microbiology, Bethesda, Maryland, pages

and by Nishida et al., ibid, 236-243 (1970).

The preparation of various 7 -Ca-amino-arylacetamido]- desacetoxy cephalosporanic acids and the corresponding unsubstituted or substi- compounds in which aryl represents for example, tuted phenyl or 2- or 3-thienyl is described, 1,054,806 and in British Specifications 985,747, 1,017,624,

1,123,333, in Belgian patent 696, 026 (Farmdoc 294,494).in

U.s. patents 3,311,621, 3,352,858, 3,489,750, 3,489,751, 16871/66 3,489,752 and 3,518,260, in Japanese patent (Farmdoc 23,231), by Spencer et al., J. Med. Chem., 9 and by Kurita et al., J.Antibiotics (Tokyo) 746-750 (1966)

12/ 243-249 (1966) and see also U.S. patent 3,485,819.

Netherlands patents 68/11676 (Farmdoc 36,349) and

68/12382 (Farmdoc 36,496) and U.S. patents 3,489,750 and

3,489,751 disclose ring-substituted cephaloglycins.

Various 7-(a-amino-arylacetamidolcephalosporins in

which one hydrogen of the a-amino group is replaced by a

carbonyl group which is attached in turn to another moiety

have been reported. The earliest were the cephaloglycin common and cephalexin precursors in which use was made of a illustrated peptide blocking group such as carbobenzyloxy as

3 4635.29

(A

37,950 72 by U.S. patent 3,364,212, Belgian patent 675,298 (Farmdoc

22,206), South African patent 67/1260 (Farmdoc 28,654) Related com- and Belgian patent 696,026 (Farmdoc 29,494). and 3,311,621 pounds include those of U.S. patents 3,303,193 and 3,518,260.

C Various cephalosporins,including cephalosporin reacted with on occasion but not cephaloglycin, have been compounds nucleophilic, aromatic mercaptans to produce

having the structure

S

Acyl NH CH CH CH2 I NI I C CH2 S- Ar

C I COOH

In U.S. patent 3,278,531 Ar is phenyl or certain sub- stituted phenyls or certain aromatic heterocyclic rings named, for example, in column 5. Similar nucleophiles,

e.g. 2-mercaptopyrimidines, are disclosed in U.S. 3,261,832

and Great Britain 1,101,422 and U.S. 3,479,350 and U.S.

3,502,665 all issued to Glaxo. A parallel disclosure is

found in Great Britain 1,109,525 to Ciba, e.g. in definition

for R3 Additional nucleophiles of this type were

disclosed by Fujisawa in Belgium 714,518 (Farmdoc 35,307; 4 4635,29

37.950 In. Africa 2695/68), in canada Netherlands 68/06129 and South Great Britain 1,187,323 818,501 (Fardoc 38,845), in 67/14888), in U.S. 3,530,123 (Farmdoc 31,936; Netherlands 34,328; Netherlands and in U.S. 3,516,997 (Farmdoc compound named cefazolin, 68/05179 which includes the sidechain on the 7-amino which has a tetrazolylacetyl group at the group and a 5-methyl-thiadiazolylthiomethyl at some length in the scientific 3-position and is described Agents and Chemotherapy literature, e.g. in Antimicrobial Bethesda, Maryland 1969, American Society for Microbiology, 23(3)1 at pages 236 243 and in J. Antibiotics (Japan) 131-148 (1970).

the structure Various cephalosporins having

S Acyl NH CH--CH CH2

C- N C CH2 S alkyl 0

I COOH

sidechains including in which acyl represents various described in some of the ca-aino-phenylacetyl have been (Farmdoc 41,619) above and by Glaxo in Belgium 734,532 and in Belgium 734,533 (Farmdoc 41,620) Cephalosporins having the structure

Acyl NH CH- CH C 2

0 1 C N C CH2 X

COOH

M 4635.,29

37, 950 /72 S 0 II II where X includes S C and S C are disclosed in some of the above and in U.S. 2,239,515; 3,239,516; 3,243,435; 3,258,461; 3,431,259 and 3,446,803.

Related publications in the scientific literature

include J. Med. Chem. 8, 174-181 (1965) and J. Chem. Soc. (London) 1595-1605 (1965) 5015-5031 (1965) and 1959-1963 (1967).

More recently, replacement of the 3-acetoxy group of a cephalosporin by various heterocyclic thiols has been

disclosed in U.S. 3,563,983 and in Netherlands 70/05519 8 (Farmdoc 0,188R) where the sidechains were, for example,

7-a-aminophenylacetamido and typical heterocyclic thiols were 2-methyl-1,3, 4 -thiadiazole-5-thiol and 1-methyl-1,2,3, 4

The present invention provides a compound of the for- mula

0 S II Ar CH c NH CH CH CH HN N NH2 c N C CH2 S C N o I c0 I COOH H Formula I

wherein Ar represents phenyl, 2-thienyl or 3-thienyl.having the D configuration in the sidechain and the nontoxic, pharmaceu- tically acceptable salts thereof. This compound exists primarily Sas the zwitterion.

4635,29

37,950 n The salts include the nontoxic carboxylic acid salts

thereof, including nontoxic metallic salts such as sodium,

potassium, calcium and aluminum, the ammonium salt and

substituted ammonium salts, e.g. salts such as nontoxic amines

including triethylamine, procaine, diben- as trialkylamines,

zylamine, N-benzyl-beta-phenethylamlne, l-ephenamine, N,N'-

dibenzylethylenediamine, dehydroabietylamine, N,N'-bis-dehy-

droabietylethylenediamine, N-(lower)-alkylpiperidine, e.g.,

N-ethylpiperidine and other amines which have been used to

form salts with benzylpenicillin; and the nontoxic acid addition

salts thereof the amines salts) including the mineral

acid addition salts such as the hydrochloride, hydrobromide,

hydroiodide, sulfate, sulfamate and phosphate and the organic

acid addition salts such as the maleate, acetate, citrate,

oxalate, succinate, benzoate, tartrate, fumarate, malate, man-

delate, ascorbate and the like.

4635.29

37,950 in This invention also provides a compound of

the formula

FN-- CH? NN H H II C-CH S C N I H COOH

Formula II

and the salts and esters thereof as defined herein. The invention further provides a process

involving the use of a compound of Formula II for preparing a compound having the formula 0 S

Ar CH C NH CH-- H CH2 N

NH C N C CH 2 S C N 2

0 C COOH H

Formula I

wherein Ar represents phenyl, 2-thienyl'or 3-thienyl

having the D configuration in the sidechain and the

nontoxic pharmaceutically acceptable salts thereof. Said process is characterized by reacting a compound

having the formula

4635.29

37,950 72 S

H2 N CH CH CH HN N I 1 I 1 c N C CH 2 S C N

C C COOH H

Formula II

or a salt or easily hydrolyzed ester thereof including those of U.S. patent 3,284,451 and any of the silyl esters described in U.S. patent 3,249,622 for use with 7-aminopenicillanic

acid and used in Great Britain 1,073,530 with a particular acid or its functional equivalent as an acylating agent for a primary amino group. After coupling, the blocking group is removed to give the dsaired product. Said acid has the formula

Ar CH COOH I NHB

wherein Ar is as defined above and B represents a blocking group of the type used either in peptide syntheses or in any of the numerous synthesis of a-aminobenzylpenicillin from 2 -phenylglycine. Particularly valuable blocking groups are a proton, as in the compound of the formula

0 CH C Cl

NH2'HC1

4 635.29

37,950 72 or a B-diketon as in Great Britain 1,123,333, methyl

acetoacetate, in which case the acid containing the blocked

amino group is preferably converted to a mixed anhydride,

as with ethyl chloroformate, before reaction with compound

II or a salt thereof to form the desired product I after

acid cleavage.

Further to the discussion above of blocking groups

used on the free amino group of the sidechain acid during its

coupling with compound II, the blocking group is then re-

moved to form the products of the present invention, e.g. the

t-butoxy-carbonyl group is removed by treatment with formic

acid, the carbobenzyloxy group is removed by catalytic

hydrogenetion, the 2-hydroxy-l-naphthcarbonyl group is re- moved by acid hydrolysis and the trichloroethoxycarbonyl

group by treatment with zinc dust in glacial acetic acid.

Obviously other functionally equivalent blocking groups for

an amino group can be used and such groups are considered

within the scope of this invention.

Thus, with respect to said acid to be used to couple

with compound II, functional equivalents include the corres-

ponding acid anhydrides, including mixed anhydrides and

particularly the mixed anhydrides prepared from stronger acids such as the lower aliphatic monoesters of carbonic acid, or

alkyl and aryl sulfonic acids and of more hindered acids such

as diphenylacetic acid. In addition, an acid azide or an

active ester or thioester with p-nitrophenol, 2,4-dini-

trophenol, thiophenol, thioacetic acid) may be used or the

free acid itself may be coupled with compound II after first

reacting said free acid with N,N'-dimethylchloroformiminium

in 463529

37,950 72 chloride [cf. Great Britain 1,008,170 and Novak and Weichet,

Experientia XXI, 6 3C0, (1965)] or by the use of enzymes or of an N,N'-carbonyldiimidazole or an N,N']carbonyl-

ditriazole [cf. South African patent specification

63/2684) or a carbcdiimide reagent (especially N,N'-

dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide or

N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide; cf.

Sheehan and Hess, J. Amer. Chem. Soc., 77, 1067 (1955)], or of alkylylamine reagent [cf. R. Buijle and H.G. Viehe,

Angew. Chem. International Edition 3,582 (1964)], or of a ketenimine reagent [cf. C. L. Stevens and M.E. Mond, J. Amer. Chem. Soc., 80, (4065)1 or of an isoxazolium salt reagent [cf. R. B. Woodwar, R.A. Olofson and H. Mayer, J. Amer. Chem. Soc., 83, 1010 (1961)3. Another equivalent of the acid chloride is a corresponding azolide, an amide of the corresponding acid whose amide nitrogen is a

11 4 635.s2

37,950 72 member of an quasiaronatic fivemembered ring. containing at

least two nitrogen atoms, imidazole, pyrazole, the tri-

azoles, benzimidazole, benzotriazole and their substituted

derivatives. As an example of the general mehhod for the

preparation of an azolide, N,N'-carbonyldiimidazole is reacted

with a carboxylic acid in equimolar proportions at room

temperature in tetrahydrofuran, chloroform, dimethylformamide

or a similar inert solvent to form the carboxylic acid imida-

zolide in practically quantitative yield with liberation of

carbon dioxide and one mole of imidazole. Dicarboxylic acids

yield dimidazolide. The by-product, imidazole, precipitates

and may be separated and the imidazolide isolated, but this is

not essential. The methods for carrying out these reactions to

produce a cephalosporin and the methods used to isolate the cephalosporin so produced are well known in the art. In the treatment of bacterial infections in man, the compounds of this invention are administered orally or parenterally, in accordance with conventional procedures for antibiotic adminis- tration, in an amount of from about 5 to 200 mg./kg./day and

preferably about 5 to 20 mg./kg./day in divided dosage, e.g.,

three to four times a day. They are administered in dosage

units containing, for example, 125 or 250 or 500 mg. of active

ingredient with suitable physiologically acceptable carriers or excipients. The dosage units are in the form of liquid

preparations such as solutions or suspensions or as solids in tablets or capsules. Exactly 200 g. of 7-aminocephalosporanic acid (7-ACA) was

suspended in 500 ml. of acetone and a solution of 240 g. of 2- Stoluenesulfonic acid in 500 ml. of acetone was added in one charge. After stirring for five minutes, at room temperature, the mixture

4 63 52 9

37 950 /72 ant the bed was filtered through diatomaceous earth 'uper Cel") about washed with 150 ml. of acetone (the insoluble matter weighed and, while Then 80 ml. of water was added to the filtrate stirring, the a-toluene-sulfonate salt crystallized out after rod. The scratching on the inside of the flask with a glass minutes and suspension was stirred in an ice-salt bath for thirty filtered cold. It was washed with 2 X 200 ml. of cold acetone and air dried; yield 250 g. of salt. This 2-toluene- and sulfonate salt of 7-ACA was stirred in 2 liters of methanol The filtrate the insoluble matter filtered through "Super Cel". liters of water were was placed in a five liter 3 neck flask and 2

added. Then the pH was adjusted to 4 by the addition of con- stirred centrated ammonium hydroxide with cooling and the suspension

for one hour at 0° C. The product was collected by filtration and

washed with 2 X 100 ml. H20 and 3 X 1 liter acetone (room

temperature). After air drying, the yield of 7-ACA was 145 g.

Reference: Glaxo, British Patent 1,104,938 (1968).

The following examples are given in illustration of,

but not in limitation of, the present invention. All tempera-

tures are in degrees Centigrade. 7-Aminocephalosporanic acid is

abbreviated as 7-ACA and methyl isobutyl ketone as MIBK.

"Skellysolve B" is a petroleum ether fraction of B.P. 60-68" C. consisting essentially of n-hexane.

Description of the Preferred Embodiments

Example 1

Sodium D-a-[l-carbomethoxvyropen-2-yl)-aminol-phenl.k

acetate

Lit. ref. E. Dane, F. Oreis, P. Konrad, T. Dockner,

Angew. Chem. Intern. Ed. Ens. 1, 658 (1962); F. Dane and T. Dockner

Angew. Chem. I6, 342 (1964); Spencer, Flynn, Roeske, Sin and

4635.29

37.950 72 Chavette., J. Med. Chem., 746-50 (1966); U.S. Patent 3,496,171

(Lill). To a well stirred rixture of 40 g. (1 mole) of NaOH in ml. of H2 and one liter of benzene was added 151.6 (1 mole) of D-(-)-phenylglycine. The mixture was held at about 550 C. for minutes and then with vigorous stirring 116 g. (1 mole) of m&thyl acetoacetate was added and the mixture stirred and heated at reflux until no more water was collected in the Dean Stark trap. Next one liter of acetone was added with the heat removed and then the slurry was cooled and stirred 30 minutes in an ice-salt bath. The product was collected by filtration, washed well with copious amounts of acetone and air dried. Yield was 191 dec. pt.

2520 [al 20 C. +2070 (C 1

Synthesis of potassium 1,2,3-triazole-5-thiolate

SH SK

0C-N=C=S CH2'2 NHC00 N N

163.19 42.04 205.24 H H 101.13 139.23

The synthesis of the thiol was accomplished by a procedure essentially identical to that described in the literature Goerdlei

arl G. GnadChem. Ber. 00, 1618 (1966)] 5-Benzamido-l.2.3-thiadiazole To a stirred solution of benzoylisothiocyanate (50.6 p., 310 mmoles) in commercial anhydrous ether (400 maintained at

0 and in a nitrogen atmosphere, was added dropwise with vigorous stirring, 0.685 N ethereal diazomethane (453 ml., 310 mmoles). When

'he addition was completed, the mixt're was stirred for 1 hour at 0O, the solid was collected by filtration and dried in vacuo. The

4 63 5,.2 9

37,950172 melting point of the crude material (23.3 thus obtained was

observed somewhere in the region 232 to 257°. Goerdler reported M.p.

A small second crop (2.1 was ob- 267° for the pure material.

tained by evaporation of the mother liquor in vacuo. The total

yield was therefore

1.2.3-Triazole-5-thiol. A solution of the above benzamido compound (8.2

mmoles) in 211 sodium hydroxide (80 ml., 160 mmoles) was heated under

reflux temperature in a nitrogen atmosphere for 24 hours. The so-

lution was cooled to 0° in ice, and concentrated hydrochloric acid

(26 ml.) was added, while a continuous stream of nitropen was passed

through the solution. The benzoic acid which precipitated was

collected by filtration; the filtrate was saturated with sodium

chloride and the additional benzoic acid which separated was

removed by filtration. The filtrate was immediately extracted

with ethyl acetate, the extract was washed with saturated salt

solution, dried over magnesium sulfate and then evaporated in

vacuo. The viscous oil which remained was immediately evapora-

tively distilled in vacuo (70-75°/0.001 mm.) to give an oil (2.84g., 70%) which solidified 52-59°; Goerdler reported

m.p. 60°) spontaneously. Potassium 1.2.3-Triazole-5-thiolate To a solution of the above thiol (2.84 28.1 mmoles)

in absolute ethanol (28 ml.) was added 1.93 R alcoholic potassium

hydroxide solution (14.5 The solution was then diluted with

anhydrous ether until crystallization of the salt was completed.

The solid was collected by filtration, washed with ether, and

dried in vacuo. The salt obtained in this manner (3.65 93%)

had m.p. 225° with decomposition.

4635,29

37.950 P2 7-Arnino-3- S(1l.2.3-triazole-5-vl)-thioineth-vll -3-ceohem-4- carboxylin acid. (II) Ten grams (0.075 mole) of 5-r-ercapto-l,2,3I-triazole potass salt was added to a stirred slurry of 19 g. (0.07 mole) of purified 7-aminocephalosPoranic acid and 5.9 g. (0.07 mole) of NaHC0., in 350 ml. of 0.1M phosphate buffer (pH 6.4) and the mixture heated and stirred at 550 C. for 3,1/2 hours under a nitrogen atmosphere. The resulting solution was cooled to 220 C. and the pH adjusted to with 40% HP04 The resulting precipitate was filtered off, washed with cold water (50 ml.) and air dried. The yield of 7-amino-31-[S- (l,2,3-triazole-5-yl)-thiomethyl)-31-cephem-k4- carboxylic acid was 8 dec. pt. 2'300 C. Ir analysis showed some decomposition of the O-lactam ring but it was used "as is" for the next step. Anal. Calcd. for C10 H11 N 2 s2 C, 38.39; H,3.. Found c, 38.m6; H, 3.78.

thiomcth 11-'5-cecihem-4I-carboxylic acid WI To a stirred suspension of 5.42 (0.02 mole) of sodium D-a-[I-carhomethoxypropen-2-yl)-amino]-phenyl acetate in 50 ml. of acetonitrile was added 2 drops of N,N-dimethylbenzylamine and the slurry cooled and stirred at -100 C. while 2.14 r. (0.02 mole) of ethyl chlorofortrate was added. After 20 min. at -100 a solution orl 6.26 g. (0.02 mole) of 7-amino-3,-[S-(l,2,3',-triazole- 5-Yl)-thior~ethyl]-3"-cephem-4-carboxylic acid, 2.8 ml. (0.02 mole) of triethylamine, 40 ml. of acetonitrile, and 40 ml. of water, precoole to 0* C. was added, all at once, with vi,-orous stirring. After at 00 C. the solution was saturated with salt (excess Nanl) for min. The organic (top) layer was separated and to it there wias adn, ml. of water and the resulting solution concentrated, in vac-ia, to a volume of about 50 ml. To this solution

I 4 635. 2 9

37,950 i72

was added a solution of 70 ml. of methyl isobutyl ketone

(KIBK) and 8 ml. of 90A formic acid. The mixture was first

shaken and then stirred at 0° C. for 3 hours. The aqueous

phase was then separated and then stirred 30 min. with a

fresh 70 ml. portion of YiIBK at 0° C. and separated again.

The aqueous phase was then concentrated to near dryness

at 20° C. under reduced pressure. The residual oil was

then triturated with acetonitrile until solid. The solids

were collected by filtration, air dried, and then vacuum

dried over P205 for 14 hours. The yield was 2.45 g. of

7-(D-a-amino-a-phenyl-acetamido3-3-[S-(1,2,3-triazole-5-yl)-

thiomethyl)-3-cephem-4-carboxylic acid. This material by

ir showed evidence of phenylglycine and starting II above

in it as impurities. These were removed by dissolving

the material in 25 ml. of water, filtering adjusting the

pH to 1.5 with 40$ H 3 PO 4 adding 1 g. of "Darko KB"

decolorizing carbon, stirring 10 min., filtering through

diatomaceous earth ("Dicalite") filter aid and adjusting the

pH to 3.5 with solid NaHCO 3 ("Darko" and "Dicaliter" are

Registered Trade Marks.) Next, the clear white solution

was concentrated to a volume of about 15 m. and the white

precipitate collected by filtration, air dried and then

vacuum dried over P 2 0 5 The amorphous, water soluble S 1 product, 7-(D-4-Amino-~-phenylacetamido)-3-C ,2,3-

thiazole-5-yl)-thiomethyl3-3-cephem-4-carboxylic acid, and which weighed 500 mg, gave a completely consistent ir

nmr spectra and showed no evidence of any significant

contamination. Elemental analysis showed 6.15A water by

the Karl Fischer method and 1.61% sodium as residue.

4 635.29

37,95072 of the It is important to note that the conversion proceed thiol is knowin to berzmz-do thiaelazole to the triazole and G. Onad, Chem. Bcr. Goerdler via 5-amino-1,2,-thiadiazole 1618 (1966).

N N OH OH

N NHC0 ~S NH2 H

S- OH 87% N2H

be prepared by an alternative roite, 5-Amino-1,2,3-thiadiazole can and R. Slack, J. Chem. Soc. not involving diazonethane [D.L. Pain

5166 (1965)] 0

IN-CHF2 CHO H 2NNHCO Et 2

SOC12 4

S \Ifl

2 77%

4 68 29

37. 9 0 72 This sample of 7-[D-(a-amino-a-phenylacetamido)]-3- hyll-3-cephem-4-carboxylic acid 3 -triazole-5-l)-thimth in water at 2 mTm./ml. followed (called New Compound) after solution to exhibit the following by dilution with Nutrient Broth was found in mcg./ml. versus the Minimum Inhibitory Concentrations incubation indicated microorganisms as determined by overnight two old, orally absorbed at 370 C. by Tube Dilution. Results with

cephalosporins are also given. Table 1

M.I.C. in mcg./ml,

New Ceha- Cephalo- Organism lexin plvcin

A9585 0.08 0.16 0.04 D. pneumoniae 5% serum* A9604 0.04. 0.16 0.04 Str. Pyogenes 5% serum* A9537 0.3 0.6 0.6 S. aureus Smith A9537 0.25 1.3 1.3 S. aureus Smith 50% serum 2 1.3 S. aureus BX1633-2 A9606 0.6 at 10-3 dil'n

BX1633-2 A9606 1 4 S. aureu at 10 dil'n

S. aureus meth.- A15097 2 resistant 2 0.3 Sal. enteritidis A9531 40.25 A15119 0.5 8 1 E. coli Juhl A9675 2 16 4 E. coli A9977 00.2 8 0.6 K. pneumoniae A15130 1 16 1 K. pneumoniae A9900 i0.2 8 0.6 Pr. mirabilis A15153 16 125 63 Pr. morganii A9843A >125 >125 >125 P. aeruginosa A20019 125 125 >125 Ser. marcescens

50% Nutrient Broth 45% Antibiotic Assay Broth

4635.29

Blood levels in h se after oral

administration were determined with the following results:

C-CCONHii H CHR 2 NH 2 0 COOH Blood Level in mcq./ml. Dose 0.5 1 2

mgm./kg. Hours after administration

HN-- N 100 6.6

S- N 100 4.1 5.74 2.8 2.1

C/ H

49.9

33.2 18. 1.73 H (cephalexin)-monohydrate

Examole 2

Sodium 7-D-(c-amino-c-ohentlacetamido)]-3-So-(1,2,3- -3-ceohem-4-carboxylate.

To a stirred aqueous suspension of the zwitterionic form

of (-amino--phenylacetamido) [S-(1,2,3-triazole- is 5-yl)-methyl]-3-cephem-4-carboxylic acid (0.8 mmole) until added 1 N acueous sodium hydroxide at room temperature

a clear solution (pH 10.8) is obtained. This solution is

immediately freeze-dried to give impure, solid sodium

[-amino-sphenylacetamido)]

[-(l,2,3-triazol-5- l at e yl)-thiomethyl -3-cephem-4-carboxy o 4635,29

37.950 ft Example 3

7-D-(-)-a-Amino-a-(3-thienyl)-acetamido)-3-[S-(1.2.3 cthi~] -'1i -t 'box Ic acid

To a stirred suspension of 16 g. (0.05 mole) of 7-

amino-3-lS-(1,2,3-triazole-5-yl)-thiomethyll-3-cephem-4-

carboxylic acid, in 150 ml. of methylene chloride was added 13.5 ml. (0.092 mole) of triethylamine, 15 ml. (0.118 mole) of N,N-dimethylaniline and, with cooling (70 C. to 150 19.1 ml.

(0.15 mole) of trimethylchlorosilane. After 15 min. cooling,

the mixture was refluxed for 25 min., cooled to 50 C. and 13 g. (0.061 mole) of D-(-)-a-amino-a-(3-thienyl)-acetylchloride hydrochloride added. The mixture was stirred at 100 C. to 120 C. for 1 hour and then 150 ml. of water was added and, after 15 min. stirring, the mixture was filtered, the aq phase separated, the pH adjusted to 2 with 20% NaOH, carbon (10 g. of "Darco KB")

added and, after 15 min. stirring, the mixture was filtered. The pale yellow solution was layered with ether (150 mL) and, with good stirring, the pH wras adjusted to 4 with 20% NaOH. The aq,phase was separated and filtered and 50 ml. of aceto- nitrile added to it. Scratching or seeding induced crystal- lization and gave 4.5 g. of water washed and vacuum dried 7-[D-(-)-a-amino-a-(3-thienyl)-acetamidol-3-[S-(1,2,m- triazole-5-yl)-thiomethyll--cephem-4-carboxylic acid; dec. pt. >1500 C. (indefinite). 'Thle ir and nmr were consistent for the desired structure.. Anal. Caled for C16 H 16 N6 0 S.H 20; C, 40 .88; H, 3.86;

N, 17.87. Found: C, 40.21; H, 3.87; N, 17.6x.

4 63 5.2 9

37,950 1"l Example 4

B.2 t~l her~-cabaX~lic acid

Following the exact same procedure as in Example3 except using D- -a-aminoa(2-thienyl)acetylchloride hydro- chloride, gave 6.01 dec. pt. >1500 C. (indefinite).

The ir and nmr spectra were consistent with the desired structure. Anal. Calcd. for C1 0 16 N6 0j4S&H 2 O: c, 40.88,

H, 3.86; N, 17.87; K.F. H2o0 3.87. Found: C, 39.60; H, '3.89; N, 16.90; K.F. H2 0, 5.43.

4635,29

37,95002 invention are as follows: The claims defining the

having the 1. A process for preparing a compound

formula

0 S II CH2 HN N C NH CH CH Ar CH I I i. II N C CH2 S- C N NH2 C C 0 I COOH H

Formula I

or 3-thienyl having wherein Ar represents phenyl, 2-thienyl the nontoxic, the D configuration in the sidechain and characterized by pharmaceutically acceptable salt, thereof reacting a compound having the formula

S

CH2 HN N H 2N CH I 11 C CH 2 S N C II 0 C H I COOH

Formula II

defined, with an or salts or ester thereof, as hereinbefore acylating agent for acid or its functional equivalent as an

4635.29

Claims (1)

1. 37.950 n2 the primary amino group having the formula

   Ar CH COOH

   NHB

   wherein Ar is as defined above and B represents a blocking

   group,as hereinbefore defined,and thereafter removing said blocking group.

   2. The process of claim 1 wherein the blocking

   group is t-butoxy-carbonyl and is removed with formic acid.

   3. The process of claim 1 wherein the blocking group is carbobenzyloxy and is removed by catalytic hydrogenation.

   4. The process of claim 1 wherein the blocking group

   is 2-hydroxy-l-napthcarbonyl and is removed by acid hydrolysis.

   The process of claim 1 wherein the blocking group is trichloroethoxycarbonyl and is removed with zinc dust in

   glacial acetic acid.

   n 4635.29

   37,950 fn

   6. The compound having the formula

   o0

   Ar -CH C NH CH CH CH2

   NH N C-CH2-S-C N

   COOH

   Formula I 3-thienyl having wherein Ar represents phenyl, 2-thienyl or

   the D configuration in the side chain, or a nontoxic,

   pharmaceutically acceptable salt thereof.

   7. The compound of claim 6 having the D configuration

   in the sidechain of the formula 0

   Ar CH C NH CH-- CH CH 2 HN

   CH-S C N NH 2NH

   I COOH H

   wherein Ar represents phenyl, 2-thienyl or 3-thienyl. 8. The sodium or potassium salts of the compound of

   claim 6 or 7. 9. A nontoxic, pharmaceutically acceptable acid

   addition salt of the compound of claim 7.

   4 635.29

   0o 37,950 2 configuration

   [ff The compound of claim 6 having the D

   in the sidechain of the formula

   S N N SCH -NH CH CH2 C HN-- I I1 C C CH2 S C N NH2 0 C C COOH H

   thereof. or a nontoxic, pharmaceutically acceptable salt /I S The compound of claim 6 having the D configuration

   in the sidechain of the formula

   S HN N ScH NH CH- I CH I CH2 I I II NH2 C -C N c c- CH- S- C N H C00H

   or a nontoxic, pharmaceutically acceptable salt thereof.

   a3. The compound of claim 6 having the D configuration

   in the sidechain of the formula S HN N -NH CH CH CH2 HN- CH I I I I N C CH2 S C N S NH 0 H S COOH

   or a nontoxic, pharmaceutically acceptable salt thereof.

   26 4635.29

   37.950 in 13. The process of claim 1 substantially as herein

   before described with reference to the Examples.

   14. The compound of formula I defined in claim 6

   hen creoared according to the process of clai 1.

   A pharmaceutical composition comprising a

   compound or salt as claimed in anyone of claims 6 to 12

   and a carrier or excipients.

   16. A method of treating mammals for diseases caused

   by gram-positive or gram-negative bacteria which method

   comprises administering an effective amount of a compound

   or salt as claimed in anyone of claims 6 to 12 or a

   composition of claim

   17. A method of treating cattle for mastitis which method

   comprises administering an effective amount of a compound

   or salt as claimed in anyone of claims 6 to 12 or a

   composition of claim

   18. A pharmaceutrical composition substantially as

   described herewith with reference to the Examples.

   19. A method as claimed in claims 16 or 17

   substantially as described herein. The compound having the formula

   C /s\ CH 2 N H2N- N C- CH 2 S N

   OOH H

   Formula II

   and the salts or esters thereof as hereinbefore defined.

   21. The sodium salt of the compound claimed in claim

   22. The potassium salt of the compound claimed in

   4635,29 claim 21. 3 7 ,950 m 23. A process for preparing a compound as claimed in any one of claims 20-22 including the step of reacting substantially equimolar weights of 5-mercapto-l,2,3- triazole with 7-aminocephalosphoranic acid.

   A process as claimed in claim 23 wherein said

   reaction is carried out in aqueous medium.

   A process as claimed in claim 23 or claim 24 wherein said reaction is carried out at a temperature of

   about 550 C.

   26. A process as claimed in any one of claims 23-

   wherein said reaction is carried out under a nitrogen

   atmosphere.

   27. A process as claimed in any one of claims 23-

   26 wherein the 5-mercapto-l,2,3-triazole is employed in

   the form of a sodium or potassium salt.

   28. A process as claimed in claim 23 substantially

   as hereinbefore defined with reference to the examples.

   DATED: 27 May, 1975. PHILLIPS ORMONDE FITZPATRICK ATTORNEYS FOR:

   BRISTOL-MYERS COMPANY

   4635,29