IL25756A - Penicillanic acid derivatives and their preparation - Google Patents

Description

Penicillanic aeid derivatives and their preparation _ BBIS Oi*BATO: RESEARCH INSTITUTE, Ci 24562 fhis invention provides ew compounds having the general formulas 1 2 in which R is hydrogen, chloro or trifluoromethyl, S is hydrogen, chloro, trlfluoromethyl, fluoro, nitro, lower alkyl or lower alkoxy, R is lower alkyl and Z is -CI, -C-BH^, -C-OH orifes reactive functional derivatives, or the group* OeC i CH-COOfI o invention also provides non-toxic, pharmaceutically acceptable salts of those compounds in which 2 is the group (A) he term "lower" relates to both straight and branched* chain aliphatic hydrocar on and hydrocarbyloxy radicals having from one to six carbon atoms.

Those compounds in which % is the group (A), i.e. 6- 5»-lower alkyl-3*-phenylisothiazole-4*-carboxaaido/-penicillanio acid derivatives and their salts, aro synthetic penicillins which are valuable as antibacterial agents, as nutritional supplements in animal feeds, as agents for the treatment of mastitis in cattle and as therapeutic agents in poultry and animals, including man, in the treatment of infectious diseases caused by (iram-positive bacteria and especially Staphylococcus aureus.

She present invention thus provides al ernate© and improved agents for the treatment of infections caused by resistant strains of bacteria, e.g. benaylpenieillin-resistant aureus) or for the decontamination of objects bearing such organisms, e.g. hospital equipment, walls of operating rooms and the like.

She salts of the penicillins provided by the present invention include metal salts such as those of sodium, potassium, calcium and aluminum$ ammonium and substituted ammonium salts, e.g. those of non-toxic amines such as trialkylamines including triethylamine, procaine, dibenzylamine, B-benzyl-beta-phenethylarnlne, l-ephenamlne, ώ,Μ· -dibenzylethylenediamine, dehydroabietylamine, bis*dehydroabiotylethylenediaiaine, N-(lower)-alkylpiperldines, e.g. N-ethylpiperidine, and other amines which have been used to form salts ¾ith benzylpenicillln.

The remaining compounds of formula I above are intermediates in the preparation of the aforesaid new penicillins.

Preferred compounds of the formula I, in which Z is the group (A), are those in which R and H are hydrogen 3 or chloro and R is methyl. 3?he present Invention also provides a process for the preparation of compounds of formula I in which Z is the group (A), and their salts, which comprises reacting β-aainopenicillanlc acid or a salt thereof with at least on© equivalent of a reactive functional derivative of the corresponding acid of foraula I in which Z is -000Η» The term "reactive functional derivatives" denomina es^^ acid chlorides and bromides, acid anhydrides, mixed anhydrides with other acids, acid assldes, asolides and those esters aad thioosters whose alcohol moiety can readily be replaced by the 6-amino group of penicillanic acid.

Among the mixed anhydrides, those derived from the acid of formula 1 in xihich Z is -COOH and a stronger acid are particularly suitable. The stronger acid may be a lower aliphatic ester of carbonic acid or an aUfcyl- or arylsulfonio acid. Also suitable are mixed anhydrides with a sterically hindered acid, such as diphenylace ie acid.

Esters and thioesters which can be used as the reactive functional derivatives are, for example, those in which the alcoholic moiety is derived from p-nitrophenol, 2,4-dinitropheno , thiophenol or thioglyeolic acid* She azolides are amides whose amide nitrogen is a member of a quasi-aromatic five-membered ring containing at least two nitrogen atoms (e.g. imidazole, pyrazole, the triazoles, benzicid-azole, benzotriazole and their substituted derivatives). As an example of the general method for the preparation of an azolide, -carbonyldiimidaaole is reacted with a carboxylic acid in ©quimolar proportions at room temperature in tetrahydrofuran, chloroform, dimetbylforraamlde or a similar inert solvent to form the carboxylic acid imidazolide in practically quantitative yield with liberation of carbon dioxide and one mole of imidazole. £i arboxylic acids yield diiraidazolides· The by-product, Imidazol ) precipitates and may be separated and the imidazolide isolated but this is not essential. The methods for carrying out these reactions to produce a penicillin and the methods used to isolate the penicillins so produced are well known in first reacting said free acid with .φ minium chloride ef. British Specification 1,008*170 and Hovak and Welch*, Bxperientfe XXI/6, 560 (1965)./. 6-Aminopenlcillaai acid nay also be acylated by the free acid by the use of enzymes or in the presence of an jI carbonyldlimidaaole QT an carbonylditriassol© c . South African Patent Specification 63/268 » a earbodiimide reagent ^especially N,H' -dicyclohexyl-carbodiiroide, w,K'-diiso ropylcarbodiimide or £-cyclohexyl-ii·-(2-morpholinoethyl)earbodiimideI cf. Sheehan and Hess, J. Amer. Chem, Soe.22» 106 » (1955)./, an alkynylamine reagent £cf„ R.

Bui^le and B.G. Viehe, Angew, Cheia. International Edition » 582 (1964)J a ketenlmine reagent £cf. C. I. Stevens and M.S.

Honk, J. Amer. Chem. Soc. 80» 4065 (1958) or an isoxazolium salt reagent £of. R.B. Woodward, R.A. Olofson and II. Mayer, J.

Amer, Chem, Soc.8JS, 1010 {1961)J.

She acylation reaction may be conducted in an inert organic solvent or in a mixture of such solvent and water.

Suitable inert organic solvents include acetone, tetrahydrofuran, dioxane, dimethyIformamide, dim th lacetaalde, chlorinated aliphatic hydrocarbons (e.g. methylene chloride), dimethylsu-ttoxide, methyl isobutyl ketone and dialkyl ethers of ethylene glycol or d ethylene glycol. In some instances it may be desirable to add a solution of the acylating agent in a solvent such as benzene to a solution of a salt of 6»aminopenicillanic acid in an aqueous organic solvent (e.g. acetone-water) . In such an instance the reaction medium may be either one phase or two phases, depending on the relative amount of water and acetone. Vigorous stirring is, of course, preferred when using a two-phase reaction medium. ¾hile the acylating reaction may be conducted over a wide range of from about -50°C. to about +50°C, the preferred reaction temperature is froa about -5°C. to about *15°C. prepared in the manner exemplified in the examples below and especially from known anilines according to the following reaction scheme in which - represents the radical in which F and R2 have the meaning set forth above, R represents a (lower)alkyl group and X2 represents iodine or bromine : _ NaNO , CuCN _ Na, CH,CN ¾S ^ _NH2 ≤ _CN 2 -CCH2CN NH III NaNOg, Cu20 CN ONH, In another useful procedure compound III above converted to compound IV above in the following manner In an alternative method, compound I above is which is then reacted with CuCN to give compound II above.

In a variation of this procedure the starting benzonitrile, as illustrated by 2, 6-dichlorobenzonitrile, prepared in the following manner: Use may be also made of the following procedure to S0C1, HN02 Alternate routes to the desired 5-alkyl-3~aryliso-thiazole-4-car"boxyllc acids are those set forth below as illustrated by the preparation of 5-methyl-3-phenylisothiazole-4-carboxylic acid, as follows: ( LCOCl NH- CgHj-COCHgCOOEt * C6H5-C-CH2C00Et NH t * * * * * * * * * * CH,COCH, Br, III. CglLCHO — C^HcCB=CHCOCH-¾ ^→ Cr^HncCuCHn - CH-COCH, I I Br Br BuLi C^-Hj-CHO * CH^-C =C-Li —2-3 i» C6H5CH(OH)-C=C-CH5 * * * * * * * * * V. CH-.-C * * * * * * * * * CgH5COCH= CHC1 KSCN CgH5COCH =CH-SCN BuLi; CH,I * * * * * * * * * 1 CrO. CO - C a CH Ξ CHSCN 2 3 k multiple steps OOOH as above under VI 6 7 θ 9 The following examples will serve to Illustrate 11 this Invention without limiting it thereto. All melting 12 points are uncorrected and all temperatures are given in 13 degrees Centigrade. l*f 16 17 18 19 21 22 23 24 26 27 28 29 EXAMPLE 1 Benzonitrile . - The preparation of benzonitrile followed the procedure of Sandmeyer [Ber. 17, 2653 (l88 )].

A mixture of 70 g. (l.O mole) of aniline, 220 ml. of concen-trated hydrochloric acid and 800 ml. of water was diazotized with 70 g. (l.O mole) of sodium nitrite in 200 ml. of water at 0-5° C. in the usual manner. To a stirred mixture of 250 g. of copper sulfate, 280 g. potassium cyanide and 1 .5 L water was added the diazotized solution at 90° C. and stir-ring was continued for another three hours. The reaction mixture was distilled until about one-third of the contents of the flask remained. The distillate was extracted with three 200 ml. portions of ether. The ether extracts were dried over anhydrous sodium sulfate. The ether was removed by distillation and the residual oil was distilled; b.p. 185-192° C. (lit. 190 ° C); yield 80 g. (78$) of benzonitrile.

-Imino-g-phenylpropionitrile . - The preparation of β-imino-β- henylpropionitrile was carried out according to the procedure of Holzwart (J. Prakt. Chem., [2] ^9, 242). To a stirred dispersion of 72.5 g. (3 . 16 atoms) of metallic sodium in 500 ml. of toluene and 600 ml. of benzene was added dropwise a mixture of 163 g. ( 1 .58 moles) of benzonitrile and 129 g. (3 . 16 maintained. After the addition was completed, stirring and reflux-ing by external heating were continued for seven hours. After cool-ing, the precipitated sodium salt was collected by filtration. The salt was treated with water to give the free base. Yield 130 g. (57 ) of p-imino- -phenylpropionitrile. An analytical sample was recrystallized from petroleum ether, m.p. 86-87° C. (lit. 86* C.) 250 mt 11> 350) , 29 ταμ (£ 11, 220) . 3490, 3390, 2210, 1640, 1593, 1575, 1500, 1450, 1410, 785, 710 cm"1.

Anal. Calc'd. for CgHgNgt C, 74. 97; H, 4.59; , 19-42. Pound; C, 74.97, 75.28; H, 5 .36, 5.32; N, 19. 15, 19.51. g-Imino-g-Phenylthioproplonamide . - p-Imino-^-phenylthio-propionamide was prepared according to the procedure of J. Goerdeler and H. ¥. Pohland [Ber. ^4, 2950 (1961) ] . Liquid hydrogen sulfide (6 ml.) and 0.05 g. of potassium hydroxide were placed in a 100 ml. ampoule cooled with Dry Ice-acetone mixture at -60® C. and a solu-tion of 5.8 g. (0.04 mole) p-imlno^-phenylpropionitrile in 35 ml. of methylene chloride was added. The ampoule was sealed and allowed to stand at room temperature for a week. The precipitate of β-ΙιηΙηο-β-ρηβ^ΙΐηΙορΓορΙοηβπιΙάβ which separated was collected by filtration and washed with ether. Yield 4.0 g. (57$) . The an-alytical sample was recrystallized from benzene. M.p. 169-17I0 C. (lit. 174θ c). 237 m (€ 9,700) , 295 ημ (£ 8,300) , 354 ταμ (£ 19,400). i^013480, 3400, 3320, l6l0, 1550, 1500, 1475, 1390, 1280 (broad), 1130 (broad), 940, 775 cm"1.

Anal. Calc'd. for CgH1()N2Ss C, 60.66; H, 5 - 66j N, 15 -72. Pound: C, 60.77, 60.93; H, 5 .36, 5.5Ο; N, 15.19, 15. 10.

-Amino-3-Phenyllsothlazole. - 5~Amino-3"Phenylisothia-zole was prepared according to the procedure of J. Goerdeler and H. W. Pohland [Ber. ^4, 2950 (1961) ] . To a stirred mixture of (0.36 mole) of potassium carbonate in I.65 L of ether was added a solution of 42.3 g. (0.37 mole) of iodine at 35° C. Stirring and refluxing were continued for another four hours. The reaction mix-ture was poured into 5 L of ice water. The ethereal layer was dried over anhydrous sodium sulfate and evaporated to dryness to give 5-amino-3-phenylisothlazole. Yield 21 g. (71#)J m.p. l6o° C. (lit. I63e C). ^ξ^ ^ χαμ (£6,000), 2βθ (68,900). V^01 3480, 3300, 3180, 1615, 1530, 1413, 1395, 775 cm"1.

Anal, calc'd. for CgHgN2S. C, 6l.31; H, 4.58; N, 15.90. Found: C, 61.55, 61.69; H, 4.32, 4.11; N, 15.6l, 15.67.

-Amlno-4-bromo-3-phenylisothiazole. - The preparation of 5~amino-4-bromo-3-phenylisothiazole followed the procedure used for 5-amino-4-bromo-3-methylisothiazole by D. Buttimore et al. (J. Chem. Soe. 1963. 2032). A solution of 30 g. (0.17 mole) of 5-amino-3-phenylisothiazole in 100 ml. of acetic acid and 300 ml. of benzene was bromlnated with 28 g. (0.175 mole) of bromine at 5-10° C. The separated hydrobromide was collected by filtration and stirred with excess 2 N sodium carbonate solution for three hours at room tem-perature to give the free base, 5~amino-4-bromo-3~phenylisothiazole, which was recrystallized from ligroin. Yield 35 g. (8l#); m.p. 80-82° C. /) *?H 236 m/x (612,500), 262 m (£12,300). J™ 3440, 3300, 298Ο, 1610, 130, 1450, 1420, 1385, 1280, 920, 770, 705 cm"1.

Anal. Calc'd. for C^Brl^S: C, 42.36; H, 2.77; N, 10.9. Pound: C, 43.19, 43.10; H, 2.81, 2.6l; N, 10.71, 11.14. 4-Bromo-3-phenyllsothlazole . - The preparation of 4-bromo-3-phenylisothiazole followed the procedure used for 4-bromo-3-methyl-isothiazole by D. Buttimore et al. (J. Chem. Soc. 1963, 2032). A viscous solution of 57 g. (0.224 mole) of 5-amino-4-bromo-3-phenyl-isothiazole in 100 ml. of concentrated sulfuric acid and 110 ml. of of sodium nitrite in 60 ml. water at 0-5° C. in the usual manner. The dlazotized solution was added dropwiae to a stirred suspension of 2 g. cuprous oxide in 240 ml. hypophosphorous acid with cooling by water. Two additional 7 g. portions of cuprous oxide were added to the reaction mixture and stirring was continued for another one hour at room temperature. The reaction mixture was filtered with a "Dicalite" precoated filter and the residue was washed with 500 ml. ether. The filtrate was diluted with an equal volume of water and extracted with three 300 ml. portions of ether. The combined ethereal solution was washed with two 50 ml. portions of water and dried over anhydrous sodium sulfate. The ether was distilled off to give an oily residue, which was recrystallized from petroleum ether using active carbon. Yield 39 G. (73$) of -bromo -3-phenyl-isothiazole. M.p. 3- ^° C. f!ίΗ 27 .5 n¾u (€ 11, 800 ). j JJ* 1480, 1 5, 1390, 1200, 9^0 cm"1.

Anal. Calc«d for C^gBrNS: C, 45.01; H, 2.53; N, 5.80. Pound: C, 4 .36, 45.46; H, 2.65, 2.35; N, 6.8Ο, 7.59- 4-Cyano-3-phenylisothiazole. - A mixture of 21 g. (Ο .Ο87 mole) 4-bromo-3-phenylisothlazole, 9.0 g. (0. 1 mole) cuprous cyan-ide and 200 ml. γ-picoline was refluxed for 15 hours. The reaction mixture was poured into 200 ml. ice water, acidified with concen-trated hydrochloric acid and extracted with three 100 ml. portions of ether. The ethereal solution was washed with 20 ml. portions of water, dried over anhydrous sodium sulfate and evaporated to give an oil, which was crystallized from ligroin. Yield 12 g. (7^) of 4-cyano-3-phenylisothiazole. M.p. 5^-55° C. ^?5H 256.5 mji (611,800) , 273 τψ (£ 10,500) . j/m™ax 2280, 1490, 1450, 1410, 13^0, 1010, 875, 775 cm"1. (ln CCl^, 60 Mc) 2. l6 (multiplet, 5H) , Ο.87 (singlet, 1 Anal Calc'd for C HN S: C 64. 4 H .2 N 1 .04. 4-Cyano-5-methyl-3-phenylisothiazole. - The preparation followed the procedure used for 4-bromo-3, 5-dimethylisothiazole by D. Buttimore et al. (J. Chem. Soc . 1 65. 2032). To a stirred solu-tion of 18.6 g: (0.1 mole) 4-cyano-3-phenylisothiazole in 100 ml. dry tetrahydrofuran was added n-butyl lithium solution from 2.7 g. (0.4 atom) lithium, ° g. (0.22 mole) n-butylbromide and 100 ml. of dry ether at -70° C. After the addition was completed, stirring was continued for another one hour at -70° C. and then 28 g. (0.2 mole) methyl iodide was added dropwise to the chilled reaction mix-ture at the rate that the temperature did not rise above -65° C.

Stirring was continued for another two hours at -60° C. and the temperature was allowed to rise to room temperature. The reaction mixture was treated with 100 ml. water and acidified with concen-trated hydrochloric acid. The water layer was extracted with 100 ml. ether. The combined ethereal solution was washed with 20 ml. por-tions of water, dried over anhydrous sodium sulfate and distilled to leave a brown viscous oil which was fractionated in vacuo . The fraction boiling at 110-125° C/0.2 mm. (8.3 g.) was collected. The oil was crystallized from ligroin to give 4.4 g. of crystals of 4-cyano-5-methyl-3-p enylisothiazole, m.p. 72-73° C. ^^°H 230 mu (€ 13,600) , 262 πμι (£11, 900) . 2280, 1510, l46o, 1405, 1380, 790, 720 cm"1. T(in CCl^, 60 Mc) 7.26 (singlet, 3H), 2.20 (multi-plet, 5H).

Anal. Calc'd for C, 65.97; H, 4.03; N, 13.99-Found: C, 66.14, 66.04j H, 4.05, 4.37; N, 14.02.

-Methyl-5-phenylisothiazole-4-carboxamlde. - A solution of 100 mg. (0.5 m.mole) of 4-cyano-5-methyl-3-phenylisothiazole in 2 ml. of concentrated sulfuric acid was warmed at 60-70° C. on a water bath for three hours. The reaction mixture was poured onto washed with water and recrystallized from benzene. Yield 80 mg. (73$) of 5-methyl-3-phenyllsothiazole-4-carboxamide, m.p. 194-196° C. in a sealed tube. r) * in itE 269.5 m 'u (£14,000). J m*et x 3420, 3200, l640, I525, 1440, 1385, I36O, 1110 cm"1.

Anal. Calc . for C, 60.53; H, 4.62; N, 12.84. Pound: C, 60.54, 60.93; H, 4.97, 4.64j N, 12.92.

-Methyl-3-phenylisothiazole-4-carboxylic Acid. - A solu-tion of 3.82 g. (O.OI9 mole) of 4-cyano-5-methyl-3-phenylisothiazole in 76 ml. of concentrated sulfuric acid was warmed on a water bath at 60-70° C. for three hours. To the stirred reaction mixture was added dropwise 2 g. (0.029 mole) sodium nitrite in 5 ml. water at 0-10° C. and stirring was continued for another one hour at 10° C. and for two hours at 50-600 C. The reaction mixture was poured onto 200 g. crushed ice and the separated crystals were collected by filtration. Yield 3.3 g. (80#) of 5-methyl-3-phenylisothiazole-4-carboxylic acid. The analytical sample was recrystallized from benzene-ligroin (2:3), m.p. 151-153° C. ' IETl*clX0H 263 m*u (610,600) .

J?maxi lg75, 1500, 1440, 1370, 1300, 950 cm"1. T(±n CD,C0CD,2, 60 Mc) 7.84 (singlet, 3H), 3.84 (singlet, broad, 1H), 3.0 (multiplet, 5H) .

Anal. Calc'd. for C^E^O^ C, 60.25; H, 4.14; N, 6.39. Pound: C, 60.51, 66.57; H, 4.07, 4.42; N, 6„4l.

-Methyl-3-phenylisothlazole-4-carbonyl Chloride . - A mix-ture of 3.2 g. (0.0147 mole) 5-methyl-3-phenylisothiazole-4-carbox-ylic acid and 5 ml. thionyl chloride was warmed on a water bath at 7Ο-8Ο0 C. for one hour. The excess thionyl chloride was removed by distillation under reduced pressure to give an oil which was dis-tilled in vacuo. Yield 3.2 g. (95$) of 5-methyl-3-phenylisothla-zole-4-carbonyl chloride, b.p. 122-125° C/0.6 mm. 1770, 1490, 1450, 1400, 1360, 1230, 1105 cm"1. penicillanate. - A solution of 3.2 g. (0.0136 mole) 5-methyl-3-phenyl isothiazole-4-carbonyl chloride in 5 ml. methylene chloride was added over a period of two minutes to a rapidly stirred solution of 3 g. (O.OI38 mole) 6-aminopenicillanic acid and 3.0 g. (0.03 mole) tri-ethylamine in 50 ml. methylene chloride at 5-10° C. The reaction mixture was stirred for one hour at 15° C. and extracted with three 50 ml. portions of water. The combined water extracts were washed with two 50 ml. portions of ether, layered with 100 ml. ethyl ace-tate and adjusted to pH 2 with 10$ hydrochloric acid. The ethyl acetate layer was separated and the water layer was again extracted with two 100 ml. portions of ethyl acetate. The combined ethyl acetate extracts were washed with 50 ml. water, dried with sodium sulfate, filtered (an additional 0 ml. ethyl acetate was used for washing the flask and filter) and treated with 4 ml. of 39$ sodium 2-ethylhexanoate in methyl isobutyl ketone. The clear solution was evaporated to a volume of about 200 ml. and the separated fine needles of sodium 6-(5-methyl-3-phenylisothiazole-4-carboxamido)-penicillanate (Crop A) were collected by filtration. A second crop (Crop B) was obtained from the filtrate by an addition of dry ether. Yield: A, 1.97 g. (33$) i B, Ο.38 g. (6$), m.p.: A, 184-190° C. (dec.) B, 180-190° C. (dec). 266.5 mu (611,200), j/Jg£ 178O, 1665, 1615, 1 40, 1410, 1330 cm"*1.

Anal. Calc'd. for C, 49.88; H, 4.11; N, 9.19. Pound: C, 50.31, 49.69 H, 4.91, 4.6l; N, 9.02, 9.23.

This compound in vitro exhibited Minimum Inhibitory Con-centrations of 0.4-0.8 meg./ml. vs. Staphylococcus aureus Smith and of 0.8 meg./ml. vs. benzylpenicillin-resistant Staphylococcus aureus BX-I633-2 and in mice versus S. aureus BX-1633-2 exhibited a CD^Q of about I.56 mgm./kg. upon intramuscular injection. This compound was 1 Alternate Preparation of 2 5- ethyl-3-phenyllsothlazole-^-carbox,yllc Acid 6 (!) (ID (in) 7 0 (VIII) k fi-Imino-fi-phenylpropionitrile (II) . This compound was prepared 6 by the procedure described above by using 700 g. (6.8 moles) 7 benzonitrile, 557 g. (15.6 moles) acetonitrile and a dispersion 8 of 512 g. (15.6 atoms) of metallic sodium in each 2 liters of 9 toluene and benzene. Yield 10 g. (52$) . (9.3 moles) p-imino-3-phenylpropionitrile in 13 liters of methylene chloride was placed in a 20-liter stainless steel bottle. The bottle was chilled at -10° C. and hydrogen sul-fide gas was bubbled into the solution until the weight in-creased by 1.68 kg. After 18.5 g. of potassium hydroxide was added, the container was stoppered tightly and allowed to stand at room temperature for three days. The reaction mixture was cooled to -10° C. and the stopper was opened. The crystals which separated were collected by filtration and washed with ether. Yield 972 g. (59$) . - ino-4-bromo-3-Phenyllsothiazole (IV) . This compound was prepared directly from -imino-p-phenylthiopropionaraide without the isolation of 5_amino-3-phenylisothiazole .

A suspension of 800 g. (4.5 moles) thioamide in 7.5 liters of ethanol was stirred and 2100 g. (13.5 moles) of bro-mine was added dropwise at about 10° C. over a period of two hours. After the addition was completed, the stirring was con-tinued for one hour. The product, 5~amino-4-bromo-3~phenyliso-thiazole, separated as the hydrobromide, which was collected by filtration and washed with 3 liters of cold ethanol. The hydro-bromide (1059 g.) was suspended in 5 liters of water and to the stirred suspension was added 5 liters of 10 sodium carbonate solution. Stirring for two hours at room temperature changed it into the free base, which was collected by filtration, washed with water and air-dried. Yield 723 g. (62$) . 4-Bromo-3-Phenylisothiazole (V) . Deamination of 5-amino-4-bromo-3~phenylisothiazole to 4-bromo-3-phenylisothiazole was carried out as described above.

-Amino-4-bromo-3-phenylisothiazole (400 g., 1.57 concentrated phosphoric acid was dlazotized with 120 g. (1.75 moles) of sodium nitrite in 300 ml. water at 0° C. The dlazotized solution was treated with a total of 23.5 g. cuprous oxide in 1.9 liters of hypophosphorous acid to give 2 g. (68$) of -bromo-3~phenylisothiazole .

-Bromo-5-methyl-3-phenylisothiazole (VI) . In the previous pro-cedure the introduction of the 5~methyl group to the isothiazole ring was made after the conversion of the 4-bromo group into the 4-cyano group. The sequence was reversed in the present prepa-ration.

To a stirred solution of 240 g. (l mole) of 4-bromo-3~phenylisothiazole in 500 ml. of dry tetrahydrofuran was added n-butyl lithium solution from 19.5 g. (2.8 atoms) lithium, 192 g. (1.4 moles) n-butyl bromide and 500 ml. of dry ether over a period of two hours at -70° C. After the addition was completed stirring was continued for another one hour at -70° C. Then 185 g. (1.3 mole) of methyl iodide was added dropwise to the cold solution at the rate that the temperature did not rise above -40° C. Stirring was continued for two hours at -40° C. and the temperature was allowed to rise to room temperature.

The reaction mixture was cooled to -10° C, acidified with 50 mi. of 10$ hydrochloric acid and then 1 liter of water was added. The aqueous layer was extracted three times with 200 ml. ether and the combined ethereal solution was washed with water and dried over anhydrous sodium sulfate. The solvent was re-moved and the residue was distilled under diminished pressure. The fraction boiling at IO5-15O0 C./0.5 mm. was collected and allowed to stand at room temperature. The crystals which sepa-rated were filtered to give 65.9 g. of the product. An addi- filtered oil by crystallization from petroleum ether. Total yield 105.5 g. (4l#) . M.p. 44-45° C. (recrystallized from petroleum ether), λ 1 273.5 % (612,800) 1490, l440, 1390, 1345, 1020, 910, 770 cm"1.

Anal. Calc'd for C-^HgBrNS: C, 47.25; H, 3.17; N, 5.51. Found: C, 47.69, 47.51; H, 3.l6, 3.45; N, 6.07. 4-Cyano-5-methyl-3-phenylisothiazole (VII) . A mixture of 101.6 g. (0.4 mole) of 4-bromo-5-methyl-3-phenylisothiazole, 4 g. (0.48 mole) of cuprous cyanide and 300 ml. γ-picoline was refluxed overnight. The reaction mixture was poured into 2 liters of ice water, acidified with 300 ml. of concentrated hydrochloric acid and filtered with a "Dicalite" (diatomaceous earth) precoated filter. The filtrate was extracted with three 700 ml. portions of ether. The ethereal extracts were washed with water, dried over anhydrous sodium sulfate and evaporated to give an oil, which was crystallized from ligroin. Yield 48.5 g. (6¾¾).

-Methyl-5-phenyllsothiazole-4-carboxylic Acid (VIII) . The preparation of the isothiazole acid was carried out by the pro-cedure described above. 4-Cyano-5-methyl-3-phenylisothiazole (38.5 g., O.19 mole) was subjected to heating with 200 ml. of concentrated sulfuric acid at 70° C. followed by treating with 14 g. of sodium nitrite in 30 ml. water at 0° C. The product weighed 34.8 g. (87^). The IR spectrum was quite different from that reported previously, although the UV spectrum was exactly the same. M.p. 151-153° C. Λ 263 % (el0,500). max 1730, 1450, 1260, 1170, 1135 cm"1. However, a recrystal-lization from carbon tetrachloride gave colorless needles which showed the same IR spectrum as that of the previous one. 1435, 1300, 945 cm . The admixture of the products before and after the recrystallization melted at 1 5-151° C.

EXAMPLE 2 Substitution in the procedure of Example 1 for the aniline used therein of one mole of 2,6-dichloroaniline produces sodium 6-[5-methyl-3-(2' *6' -dichlorophenyl)isothiazole-4-carbox-amidolpenlcillanate.

EXAMPLE 3 Substitution in the procedure of Example 1 for the aniline used therein of one mole of 2-chloroaniline produces sodium 6-[5-methyl-3~ (2· -chlorophenyl) -isothiazole-4-carbox-amido]penicillanate.

EXAMPLE 4 Substitution in the procedure of Example 1 for the aniline used therein of one mole of 4-trifluoromethylanlllne produces sodium 6-[5-methyl-3-(4l -trifluorometh lphenyl) -isothiazole-4-earboxamido]penicillanate.

EXAMPLE 5 Substitution in the procedure of Example 1 for the aniline used therein of one mole of 2,6-di(trifluoromethyl) -aniline produces sodium 6-[5-methyl-3- (2x,6» -di{trifluoromethyl}' Substitution in the procedure of Example 1 for the aniline used therein of on© mole of -roethylaniline produces sodium 6-B-methyl-3-( * ~nethylphenyl)-ieotbiazole-4-car oxamido/penicillinate.

EXAMPLE 6 (I) ( ID ( in ) salt) 3-P-Chlorophenyl-4-cyano-5-methylisothlazole (I) - A slow stream of chlorine was passed through a melted mixture of 9.2 g. ( 0.046 mole) of 4-cyano-5-methyl-3-phenylisothiazole and 3 .0 g. ( 0.013 mole) of antimony trichloride at 80-90° C. for 2 hours. The reaction mixture was dissolved in 50 ml. of chloroform, washed with water, dried with sodium sulfate and the solvent was distilled off. The residual oil was treated with ligroin and the chlorinated product was first precipitated from the solution. The product was filtered. Concentration of the filtrate gave the starting material, which was repeatedly chlorinated in the similar way. Recrystallization of the product from ligroin gave 3-p-chlorophenyl-4-cyano-5-methylisothia- zole as needles. Yield 1.15 g. ( 10#) , m.p. 114-115° C. ^χΗ "1 1.85(d), ¾ 9 cps.

Anal. Calc'd. for C^HyClNgS: C, 56.29; H, 3 .01; N, 11.94. Found: C, 5.6.02; H, 2.71; N, 11.86. 3-P-Chlorophenyl-5-methylisothiazole-4-carboxylic acid (II A mixture of 730 mg. (3 .1 m. moles) of 3-p-chlorpphenyl-.4-cyano-5-methylisothiazole and 20 ml. of cone, sulfuric acid was heated on a water bath at 70-80° C. for 4 hours. Then the mixture was cooled and a solution of 250 mg. (3 .6 m. moles) of sodium nitrite in 5 ml. of water was added dropwise with stirring at 0-7° C. The reaction mixture was stirred at room temperature for 30 minutes, then at 50-70° C. for 30 minutes and poured on to 150 g. of crushed ice. The precipitate was collected by filtration, dissolved in aqueous sodium bicarbonate and the solution was filtered. Acidi-fication with dil. hydrochloric acid gave the product, 3-p-chloro-phenyl-5-methylisothiazole-4-carboxylic acid, which was recrys-tallized from aqueous ethanol ( 1: 1) . Yield 700 mg. ( 89#) > m.p. 164-165° C. )ξ 262 mu (6 13 ,500) . ¾¾ = 0 1725 β»"1· Anal. Calc'd for CnHQClNS: C, 52.07; H, 3 .18; N, 5.52. Pound: G, 52.03 , 52.09; H, 2.87 , 3 - 13 ; N, 6.29, 6.15. 3-P-Chlorophenyl-5-methylisothiazole-4-carbonyl chlor-ide (III). - A mixture of 600 mg. ( 2.36 m. moles) of 3-p-chloro-phenyl-5-methylisothiazole-4-carboxylic acid and 5 ml. of thionyl chloride was heated at 6o-80° C. for 30 minutes on a water bath. The excess thionyl chloride was removed by distillation under reduced pressure and the residual, solid 3-p-chlorophenyl-5-meth l-isothiazole-4-carbonyl chloride, was recrystallized from benzene-petroleum ether. Yield 620 mg. (97 ) > m.p. 62-63° C. —Q 175 cm"1.

Sodium 6-(3-P-chlorophenyl-5-meth lisothiazole-4-car- (2.8 m. moles) of 6-amlnopenicillanic acid (6-APA), 700 mg. (8 m, moles) sodium bicarbonate, 4 ml. water and 50 ml. acetone was added dropwise a solution of 620 mg. (2.28 m.moles) 3-p-chloro-phenyl-5-methylisothiazole-4-carbonyl chloride in 20 ml. dry acetone at 0-2° C. The reaction mixture was stirred for 15 min-utes at 10-15° C., washed twice with 100 ml. portions of ether, layered with 100 ml. ethyl acetate and adjusted to pH 2.0 with dil. hydrochloric acid at 2° C. under stirring. The organic layer being separated, the water layer was extracted twice with 50 ml. portions of ethyl acetate. The combined ethyl acetate extracts were washed twice with 50 ml. portions of water and dried with an-hydrous sodium sulfate. The filtrate was concentrated to about 50 ml. under reduced pressure at 30° C. The concentrate was treat with 1.2 ml. of 39$ sodium 2-ethylhexanoate (SEH) . Scrubbing the flask for about 30 minutes effected crystallization. The crystals of sodium 6-(3-p-chlorophenyl-5-methylisothiazole-4-carboxamido)" penicillanate were collected by filtration, washed with ethyl acetate and dried in vacuum over phosphorus pentoxide. Yield 870 mg. (80#), m.p. 192-197° C. (dec). ¾£ 268 ιημ (€l4,400). 1770, I63 , I60 , 1405 cm"1.

Anal. Calc'd for C^H^ClN^O^ a- 2 ¾0; C, 45. ; H, 4.02; N, 8.39. Pound: C, 45.64, 45.68; H, 4.02, 4.19; N, 8.88, 8.73.

This penicillin in vitro exhibited Minimum Inhibitory Concentrations of about 0.125 meg./ml. vs. Staph, aureus Smith and of about 0.4 meg./ml. vs. benzylpenicillin-resistant . S. aureus Bx-1633-2 and in mice vs. both S. aureus Smith and S. aureus Bx-1633-2 exhibited a CD^Q of about l8 mgm./kg. upon intra-muscular injection. This penicillin was also very stable in EXAMPLE 7 (II) (in) (Sodium salt -Methyl-3-m-nitrophenyllsothlazole-4-carboxyllc acid (I) To a stirred solution of 1.32 g. (6 m.moles) of 5-methyl-3-phenyl-isothiazole-4-carboxylic acid in 10 ml. of cone, sulfuric acid was added dropwise 450 mg. (7 m. moles) of fuming nitric acid in 5 ml. of cone, sulfuric acid at -10° C. The reaction mixture was stirred for one hour at 10° C. and poured onto 100 g. of crushed ice. The precipitated 5-niethyl-3-m-nitrophenylisothiazole-4-carboxylic acid was filtered, washed with water and recrystalllzed from ethanol.

Yield 1.32 g. (800), m.p. 235-236° C. iEt°H 260 mu (£18,000). 'lmax ^max01 1680' 155°' 155^ om"le NMR ^100 Mc' ln D2° + K2C03' intl* ref . DOH): £ (cps): CH^, +295(s); ring protons, H2-562(t), H -534(q), ¾- 4 (t), H6-496(q), z JJJ^ = 12 cps; JH2H6 = JH2H4 = 2 CpS' Jh4h6 = ~° CPS* Anal. Calc'd. for σ^βΝ^Ο^β: C, 49.99; H, 3.05; N, 10.64. Pound: C, 49.56, 49.68; H, 2.93, 2.67; N, 10. 1, 10.98.

-Methyl-3-m-nltrophenylisothiazole-4-carbonyl chloride (II A mixture of 900 mg. (3 m. moles) of 5-methyl-3-m-nitrophenyliso- thiazole-4-carboxylic acid and 5 ml. of thionyl chloride was ° e e sure. The residual, solid 5-methyl-3-m-nitrophenylisothlazole-4-carbonyl chloride, was recrystallized from dry benzene. Yield 820 mg. (9W , m.p. 122-123° C i)c =0 1740 cm"1.

Sodium 6-(5methyl-3-m-nitrophenyllsothlazole-4-carbox-amido )penicillanate (III). To a stirred solution of 700 rag. (3.24 m. moles) of 6-APA, 800 mg. (9.5 m. moles) of sodium bi-carbonate, 25 ml. of water and 25 ml. of acetone was added drop-wise 820 mg. (2.9 m. moles) of 5-methyl-3-m-nitrophenylisothia-zole-4-carbonyl chloride in 30 ml. of dry benzene at 0-5° C.

The reaction mixture was stirred for 0 minutes at 10° C. washed twice with 100 ml. portions of ether, covered with 100 ml. of ethyl acetate and adjusted to pH 2.0 with dil. hydrochloric acid at 5° C. The organic layer was separated and the water layer was extracted twice with 50 ml. portions of ethyl acetate. The com-bined ethyl acetate extracts were washed twice with 50 ml. por-tions of water and dried with anhydrous sodium sulfate. The filtrate being concentrated to 50 ml. under reduced pressure at 30° C, the concentrate was treated with 1.4 ml. of 3 SEH and stirred for 3 hours at room temperature. The separated product, sodium 6-(5-raethy1-3-m-nitropheny11sothlazole-4-carboxamido )peni-cillanate, was collected by filtration, washed with ethyl acetate and dried in vacuum over phosphorus pentoxide. Yield 950 mg. (67 ), m.p. 220-225° C (dec). 265 mu (£19,300). j/jgj I760, 1625, 1600, 1400 cm"1.

Anal. Calc'd. for C19H17N 06S2Na. H20. C, 43.85; H, 4.07; N, 10.77. Found: C, 43.76, 43.76; H, 3-57, 3-49; N, 11.36, 11.43.

This penicillin in vitro exhibited Minimum Inhibitory Concentrations of about 0.4 meg./ml. vs. S. aureus Smith and of about 1.6 meg./ml. versus benzylpenicillin-resistant S. aureus EXAMPLE 8 (I) c.HgSO^ NaNO -Amino-4-bromo-3-phen llsothiazole - 5-Amino-4-bromo- 3-phenyllsothiazole, an intermediate to 4-eyano-5-methyl-3-phenyl- isothiazole, was described above in Example 1 as having m.p. 80- 82° C. This aminoisothiazole was obtained in a purer state by re- crystallizations from methylene chloride and then ligroi ; m.p. 84-85° C. A¾xH 236 mu ( 11,200), 259 ιημ 11,200).

Anal. Calc'd. for CgHyBrN S: C, 42.36j H, 2.77; N, 10.98. Pound: C, 42.38, 42.30; H, 2.69, 2.95; N, 11.02, 11.11. 3-o-Chlorophenyl-4-cyano-5-methyllsothiazole (I) - A slow stream of chlorine was assed throu h a mixture of .6 . 0.2 of antimony trichloride at 120° C. for 5 hours. The reaction mixture was dissolved in 200 ml. of chloroform, washed with 50 ml. of 10$ hydrochloric acid and then with aqueous sodium "bicarbonate solution. The chloroform solution was dried with anhydrous sodium sulfate and the solvent was evaporated under reduced pressure to afford the oil, which was dissolved in hot ligroin. The solution was treated with active carbon and allowed to cool at room tempera- ture to separate the p-chloro derivative. Yield 17 g. (26.6$).

Evaporation of the filtrate gave a mixture of p-chloro, o-chloro, polychloro (including 2,4-dichloro) and unchlorinated derivatives. The mixture was recrystallized from ligroin and 4.8 g. (7«5#) of pure o-chloro derivative, 3-o-chlorophenyl-4-cyano-5-methyliso- thiazole, was obtained, m.p. 75-76° C. ^Ϊ2Η 260.5 mu (€8,700).

NMR (100 Mcs in CCI , ref. TMS): Xv ) 7.26 (s, -CHj), 2.62 (m, phenyl ring protons).

Anal. Calc'd. for C, 56.29; H, 3.01; N, II.94. Pound: C, 56.97, 57.11; H, 3.67, 3.24; N, 11.65, 11.88. 3-Q-Chlorophen 1-5-meth 11sothlazole-4-carbox lic acid (II) A mixture of 4.0 g. ( 0.017 mole) of 3-o-chlorophenyl-4-cyano-5-methylisothiazole and 50 ml. of cone, sulfuric acid was heated at 8o° C. for 4 hours on a water bath. The mixture was chilled and 1.53 g. (0.022 mole) of sodium nitrite in 4 ml. of water was added dropwise with stirring at 0-5° C. The reaction mixture was stir-red at room temperature for 30 minutes and at 70° C. for 30 min-utes until evolution of nitrogen ceased, and poured onto crushed ice to give the product, 3-o-chlorophenyl-5-methylisothiazole-4-carboxylic acid, which was recrystallized from ethanol and water (1:1). Yield 3.7 g. (86$), m.p. 170-172° C. (in sealed tube). ^max01 1670 ^'1·" ^ma H 256 (€9,500). NMR (100 Mc; in D20 + Anal. Calcfd. for CnH3ClN02S. C, 52.07J H, 3- l8; N, 5.52. Pound: C, 51.63 , 51.46; H, 3-10, 3 - 16; N, 6.01, 5 .80. 3-o-Chlorophenyl-5-methylisothiazole-4-carbonyl chlor-lde (III). A mixture of 2.7 g. ( 0.0107 mole) of 3-0-chlorophenyl-5-methylisothiazole- -carboxylic acid and 3 ml. of thionyl chlor-ide was heated on a water bath at 80° C. for 30 minutes. The excess thionyl chloride was removed by distillation under reduced pressure and the residue was distilled in vacuum. Yield 2.55 g. (88$) of 3-o-chlorophenyl-5-methylisothiazole-4-carbonyl chloride, b.p. 1 O-I3 0 C 5 mm. ^^^id 1760 cm"1.

Sodium 6-(3-o-chlorophenyl-5-methylisothiazole-4-carbox-amldo )penlclllanate (IV). To a stirred mixture of 2.5 g. ( 0.0115 mole) of 6-APA, 2.9 g. ( 0.034 mole) of sodium bicarbonate, 80 ml. of water and 50 ml. of acetone was added dropwise a solution of 2.55 g. (Ο. ΟΟ94 mole) of 3-o-chlorophenyl-5-methylisothlazole-4-carbonyl chloride in 30 ml. of dry acetone at 5° C. and stirring was continued for 30 minutes at 10-15° C. The reaction mixture was washed twice with 100 ml. portions of ether, layered with 100 ml. of ethyl acetate and adjusted to pH 2.0 with dil. hydrochloric acid at 5° C. The ethyl acetate layer being separated, the water layer was extracted with two 50 ml. portions of ethyl acetate.

The combined ethyl acetate extracts were washed with water, dried with anhydrous sodium sulfate and evaporated to 3/4 of its volume. The concentrate was treated with 35$ SEH to give a crystalline precipitate of sodium 6-(3-o-chlorophenyl-5-methylisothiazole-4-carboxamido)penicillanate, which was collected by filtration.

When the precipitate was washed with acetone, it showed a tendency to dissolve in the solvent. The remaining precipitate "B"' ( 1.3 g.) was washed with ethyl acetate. On the other hand, to the acetone ethyl acetate. The resulting solution was concentrated slightly to give 1.6 g. of sodium 6-(3-o-chlorophenyl-5-methylisothiazole-4-carboxamido)penicilianate crystals designated as "A". Crystal-line forms "A" and "B" are different from each other in IR spectra but are almost the same in microbiological activities. "B" was also recrystallized from acetone and ethyl acetate to give 0.6 g. of crystals which show an IR spectrum identical with that of "A" . Total yield 2.2 g. (50 ).

"B": m.p. 182-188° C. (dec), j/ 1760, 1650, 1595, 1530, 1405 cm"1. 258 π (£9,100).

"A": m.p. 182-188° C. (dec). 3530, 3370, 1765, 1650, 1600, 1510, l480, 1410 cm"1. Ajjg° 257.5 πμ (69,400).

Anal. Calc'd. for H20: C, 45.55; H, 4.02; N, 8.39. Pound: C, 4 -31, 45.66; H, 3.83, 4.16; N, 8.45, 8.95.

Crystalline form "A" of this penicillin in vitro ex-hibited Minimum Inhibitory Concentrations of about 0.4 meg./ml. versus S. aureus Smith and about 0.4 meg./ml. versus benzylpeni-cillin-resistant S. aureus BXI633-2 and in mice upon intramuscular injection exhibited CD^Q'S of about 72 mgm./kg. versus S. aureus Smith and about 200 mgm./kg. versus S. aureus BXI633-2. This penicillin was also very stable in aqueous acid, having a half-life of 3.3-3.7 hours at pH 2 and 37° C.

EXAMPLE 9 (The symbol Ar represents the p-methoxyphenyl radical).

(IX) p-Methoxybenzonltrile (I) The compound was prepared from p-anlsidine according A mixture of 250 g. (2.03 moles) of p-anisidine and 670 ml. of 28# hydrochloric acid was diazotized with 150 g. (2. l8 moles) of sodium nitrite in 400 ml. of water in the usual way at 0-5° C. The reaction mixture was neutralized with solid sodium carbonate. On the other hand, cuprous chloride was prepared from 625 g. (3.9 moles) of cupric sulfate, 163 g. (3.8 moles) of sodium chloride, 133 g. (1.28 molee) of sodium bieulfite and 3 I» of water. The cuprous chloride was converted to cuprous cyanide with 325 g. (5 moles) of potassium cyanide and βθθ ml. of water. The cuprous cyanide solution was chilled to 0-5° C. and covered with 600 ml. benzene. The neutralized solution was added slowly over the v period of one hour to the cold cuprous cyanide solution with stir-ring. The stirring was continued for another two hours at room temperature and the mixture was allowed to stand overnight. The reaction mixture was warmed to 50° C. and inorganic salt was removed by filtration. The benzene layer was separated and the water layer was extracted three times with 300 ml. portions of benzene. The combined benzene extracts were washed with water and evaporated under reduced pressure to give an oily residue, which was dissolved in 500 ml. of ether. The ethereal solution was fll-tered to remove insoluble materials. The solvent was removed by distillation and the residue was distilled in vacuum to give p-methoxybenzonitrile, yield 106 g. (39#) > b.p. 105-125° C./6-7 mm., m.p. 43-46° c.

P-Imlno-P-P-methoxyphenylpropionitrile (II) The compound was prepared by the procedure used in Example 1 to prepare p-imino^-phenylpropionitrile. To a gently refluxed mixture of 125 g. (5.4 moles) of powdered sodium, 1 L toluene and 800 ml. benzene was added dropwise a solution of 22 g. nitrile in 500 ml. benzene over the period of 4 hours. The reaction mixture was stirred under reflux for another 50 hours and then allowed to stand until cool. The sodium salt which separated was collected by filtration and treated with 1.5 L water to afford the free product, p-imino-^-p-methoxyphenylpro-pionitrile. The analytical sample was recrystallized from ben-zene. Yield 175 g. (21$). M.p. 113-11 ° C. C≡N 2250 cm"1. maiH 2 m (€12,700), 295 πιμ (€18,400).

Anal. Calc'd. for C10H10N20: C, 68.95; H, 5.79; N, 16.08. Pounds C, 68.70, 68.76; H, 6.08, 5-66; N, 15-97, 16.05.

P-Imino-3-p~methoxyphenylthiopropionamlde (III) A solution of 35 g. (0.2 mole) of 3-imino-p-p-methoxy-phenylpropionitrile in 350 ml. methylene chloride was placed in 500 ml. of a pressure bottle and 300 mg. of potassium hydroxide was added. The mixture was chilled with Dry Ice-acetone. Dry hydrogen sulfide gas was passed into the chilled solution until 27 g. (0.8 mole) of the gas was absorbed. The bottle was tightly stoppered and allowed to stand for three days at room temperature. The separated bright yellow crystals of p-imino- -p-methoxyphenyl-thiopropionamide were collected by filtration and washed with ether. The analytical sample was recrystallized from methylene chloride. Yield 164 g. (80 ) . M.p. 179-180° C. A 273 πΐ (€11,000), 352 ιημ (67,900).

Anal. Calc'd. for C10H12N20S; C, 57.66; H, 5.8l; N, 13. 5. Found; C, 56.89, 57.19; H, 5.59, 5.60; N, 13-53, 13.72.

-Amino-4-bromo-3-p-methoxyphenylisothiazole (IV) The compound was prepared according to the procedure used in Example 2 to prepare 5-amino-4-bromo-3-phenylisothiazole. methoxyphenylthlopropionamide in 1.2 L of ethanol was added drop-wise 36Ο g. (2.25 moles) of bromine at 15° C. over the period of one hour. The reaction mixture was stirred for another two hours at 20° C. The separated hydrobromlde of the isothiazole deriva-tive was filtered, washed with ethanol and dried in the air. The hydrobromlde was added under stirring to 720 ml. of 2 N aqueous sodium carbonate and stirring was continued for two hours. The free product, 5~amino-4-bromo-3-p^ethoxyphenylisothiazole, was filtered, washed with water and dried. The analytical sample was crystallized from ligroin. Yield 172 g. (75$) . M.p. 109-111° C . Λ ^H 234 πΐμ (€ 12, 000) , 27 πιμ (e l8,000) .

Anal. Calc'd. for C, 42.12; H, 3. l8j N, 9.82. Found: C, 41.71, 4l .89; H, 2.77, 2.79; N, 10.50, 10.28. 4-Bromo-3-p-methoxyphenylisothlazole (V) To a stirred solution of ΐβθ g. (Ο.56 mole) of 5-amino-4-bromo-3-p-methoxyphenylisothiazole in 300 ml. of concentrated sulfuric acid and 300 ml. of concentrated phosphoric acid was added slowly a solution of 48.3 g. (0.7 mole) of sodium nitrite in 96 ml. of water at 0-5° C. The diazotized solution was added dropwise at 20° C. to a stirred mixture of 1.75 g. cuprous oxide and 750 ml. of 40# hypophosphorous acid over the period of one hour, during which four 1.75 g. portions of cuprous oxide was added. The reaction mixture was stirred another two hours at room temperature, allowed to stand overnight and filtered with a Dicalite-precoated filter. The coffee-black tarry cake was well-pressed and washed with 1 L of chloroform. The filtrate was diluted with 1 L of water and extracted with chloroform washings and with two additional 500 ml. portions of chloroform. The combined chloroform extracts were washed with water and -v- 1 which was extracted with two 500 ml. portions of hot ligroin. 2 The solvent was evaporated under reduced pressure and the residue 3 was distilled in vacuum to give 4-bromo-3-p-methoxyphenylisothia- 4 zole. Yield 92 g. (6l#) . B.p. l42-l65° C./0.8 mm. A sample of the product was crystallized from petroleum ether, m.p. 4o-4l° 6 C, Λ^Η 285 πιμ (€14,3500). ;7 Anal. Calc'd. for C^HgBrNOS: C, 44.46; H, 2.99; N, ;8 5.19. Found: C, 42.20, 43. 6; H, 2.56, 2.72; N, 5.19, 5.39. 9 4-Bromo-3-p-methoxyphenyl-5-methyllsothiazole (VI) (a) Methylation of 4-bromo-3-p-methoxyphenylisothia- 11 zole. - A solution of 8l g. (0.3 mole) of 4-bromo-3-p-methoxyphenyl 12 isothiazole in 150 ml. of dry tetrahydrofuran (THP) was chilled 13 to -7O0 C. with Dry Ice-acetone mi-xture. To the chilled solu- 14 tion was cautiously added a stirred solution of n-butyl lithium prepared from 7 g. (l mole) of lithium, 70 g. (0.51 mole) of 16 n-butyl bromide and 150 ml. of ether maintaining the temperature 17 below -650 C. over the period of 3 hours. To the mixture was 18 added 53.5 g. (0.42 mole) of methyl iodide in three portions, 19 when the temperature rose to -300 C. The reaction mixture was allowed to stand overnight and poured into 300 ml. of 2 N 21 hydrochloric acid. The ether layer was separated and the water 22 layer was extracted with two 200 ml. portions of ether. The 23 combined ethereal solution was washed with water and dried with 24 anhydrous sodium sulfate, the solvent being evaporated. The residual oil was distilled in vacuum to give 62 g. of colorless 26 oil boiling at 17O-I850 C./5 mm. The product showed two strong 27 peaks by GLC. The product was treated with hot ligroin to 28 afford crystalline 3-p-methoxyphenyl-5-methylisothiazole. 29 Yield 12.25 g. (190). M.p. 82-830 C λ 282 πΐμ (el8,700). (s, OCH-j), 3.22 (d, 3-H on phenyl ring), 2.92 (s, 4-H on iso-thlazole ring), 2.22 (d, 2-H on phenyl ring). J2H ^"^ .7 cps. · Anal. Calc'd. for C11H11NOS: C, 6 .36 H, 5.40 N, 6.82. Pound: C, 64.68, 64.76; H, 5.53, 5.53; N, 6.58, 6.40.

The mother liquor was evaporated into dryness and the residue was distilled under reduced pressure to give 4-bromo-3~ p-methoxyphenyl-5-methyllsothiazole boiling at 148-156° C./1.3 mm. Yield 28 g. (32.8#). 282 πΐμ (617,300). NMR (100 Mc; in CC1 ; TMS) : T(ppm) 7. 9 (s, -CH^), 6.19 (s, OCH^), 3.09 (d, 3-H on phenyl ring), 2.24 (d, 2-H on phenyl ring).

J2H,3H=9-7 cps* (b) Bromination of 3~p-methoxyphenyl-5-methyliso-thlazole. - To a stirred solution of 10.7 g. (0.053 mole) of 3-p-methoxyphenyl-5-methy11sothlazole and 4 g. (0.08 mole) of anhydrous sodium acetate in acetic acid was added 10.6 g.

(O.066 mole) of bromine at room temperature and stirring was continued for 3 hours. The reaction mixture was evaporated under reduced pressure. The residue was dissolved in 0 ml. of water and extracted with two 0 ml. portions of ether. The combined ethereal solution was washed with aqueous sodium bi-carbonate, dried with anhydrous sodium sulfate and evaporated into dryness. The residue was dissolved in hot ligroin. On cooling the solution gave 2.7 g. (l4#) of -bromo- -( -bromo-4-methoxyphenyl)-5-methylisothiazole melting at IO5-IO60 C.

A ?H 282 πΐμ (G16,900). NMR (100 Mc; in CCl.j TMS): 7 (ppm) 7.45 (s, -CH^), 6.04 (s, OCH^), 3.16 (d, 5~H on phenyl ring), 2.26 (q, 6-H on phenyl ring), I.98 (d, 2-H on phenyl ring).

J5H,6H= 8*3 cpS* ^Η,δΗ^2·8 Cps> J2H, 5E^° cps' Anal. Calc'd. for C H Br NOS; C 36.37 H 2.50 4.16, 5.82.

From the mother liquor, the desired 4-bromo-3-p-methoxyphenyl-5-methylisothiazole was obtained. Yield 11.7 g. (79*) . 3-p-Methoxyphenyl-5^ethylisothlazole-4-carboxylic Acid (VII) To a stirred solution of 6.5 g. (0.023 mole) of 4-bromo-3-p-methoxyphenyl-5-methylisothiazole in 40 ml. dry ether was added dropwlse at -50° C. a solution of n-butyl lithium prepared from 0.54 g. (0.078 mole) of lithium, 5.35 g. (0.039 mole) of n-butyl bromide and 30 ml. dry ether. The Dry Ice-acetone bath was removed and the temperature was allowed to rise to -30° C. The reaction mixture was poured into 50 g. of powdered dry ice. When all the dry ice was evaporated and the temperature raised to -10° C, 50 ml. of water was added and the mixture was well-shaken in a separatory funnel. The water layer was separated and acidified with dilute hydrochloric acid to give a precipi-tate of 3-p-methoxyphenyl-5-methylisothiazole-4-carboxylic acid which was crystallized from ethanol-water (l: l) . Yield 2.5 g. ( 43#) . M.p. 148-150° C. ^ c=0 1710 cm"1. 276.5 πιμ (€12,600) . NMR (100 Mc; in ^CO^ + DgO) s 8 (cps from DOH) +214 (s, 5-CH3), +87 (a, OCH^), -236 (d, 3~H on phenyl ring), -318 (d, 2-H on phenyl ring). J2H ^H = 8.9 cps.

Anal. Calc'd for c12HnN°3Si > 57.8lj H, 4.45; N, 5.62. Found? C, 57.74, 57.92; H, 4.29, 4.11; N, 5.34, 5.51. 3-P-Methoxyphenyl-5-methylisothiazole-4-carbonyl Chloride (VIII) A mixture of 3.0 g. (0.012 mole) of 3-p-methoxyphenyl-5-methylisothiazole-4-carboxylic acid and 3 ml. of thionyl The excess thionyl chloride was distilled off under reduced pressure and the residual oil was distilled JLn vacuo to give 3-p~ methoxyphenyl-5-methylisothiazole-4-carbonyl chloride boiling at 144-148° C./0.4 mm. Yield 2.9 g. (91#) . = 0 1765 cm"l e Sodium 6-(3-p-methoxyphenyl-5-methylisothiazole-4-carboxamido penicillanate (IX) To a stirred solution of 2.6 g. (0.012 mole) of 6-APA and 3 g. (O.036 mole) of sodium bicarbonate in 80 ml. of water and 50 ml. acetone was added dropwise at 5° C. a solution of 2.9 g. (0.011 mole) of 3-P"methoxyphenyl-5-methylisothiazole-4-car-bonyl chloride in 30 ml. of dry acetone. The mixture was stirred for 30 minutes at 15° C. and washed with two 100 ml. portions of ether, the washings being discarded. The reaction mixture was chilled to 5° C , layered with 100 ml. ethyl acetate and adjusted to pH 2.0 with dilute hydrochloric acid under vigorous stirring. The organic layer being separated, the aqueous layer was extracted with two 50 ml. portions of ethyl acetate. The combined ethyl acet extracts were washed with water, dried with anhydrous sodium sul-fate. The filtrate was treated with 5 ml. of 5$ SEH and concen-trated under reduced pressure below 40° C. The separated preci-pitate of sodium 6- (3-p-methoxyphenyl-5-methylisothiazole-4-carboxamido)penicillanate was collected by filtration, washed with ethyl acetate and dried in vacuo over phosphorous pentoxide.

Yield 2.6 g. (52$) . M.p. 160-1650 C. 1^ *? 1780, 16 , l6l0, 1400 cm"1. A H2° 278 πιμ (έ 12, 900) . max Anal. Calc'd. for C^H^ ^O^Na; C, 51.16» H, 4.29; N, 8.95. Pound: C, 50.90, 50.90j H, 4.70, 4.70; N, 7.91, 8.12.

This penicillin in vitro exhibited Minimum Inhibitory Concentrations of about 0.4 meg./ml. versus £>. aureus Smith and 1 Bx-1633-2 and in mice upon intramuscular injection exhibited 2 CDc^'s of about 30 mgm./kg. versus S. aureus Smith and about 3 150 mgm./kg. versus S. aureus Bx-1633-2. This penicillin was 4 also very stable in aqueous acid, having a half-life of about 4 hours at pH 2 and 37° C. 6 7 8 8 (ID 3 (Sodium Salt) 4 β 7 8 9 V 3- ( 2, -Dichlorophenyl) -- -»cyano-5-methylisothiazole (I) In Example 8, 3-o-chlorophenyl-4-cyano~5~methylisothiazole was obtained from a mixture of p-chloro, o-chloro, poly chloro (in-eluding 2, 4-dichloro) and unchlorinated derivatives. The re-maining uncrystallized residue was used as the starting material to obtain the 2, 4-dichloro derivative. A mixture of 3 g. of antimony trichloride and 13 .9 g. of the uncrystallized residue was heated at 120-130° C. and dry chlorine gas was bubbled into the mixture for 5 hours. The reaction mixture was dissolved in 50 ml. of chloroform and the chloroform solution was washed successively with 20 ml. of 10$ hydrochloric acid, 30 ml. of water and 20 ml. of 5$ sodium bicarbonate solution. The organic solution was dried with anhydrous sodium sulfate, the solvent being evaporated under reduced pressure. The residue was dis-solved in hot ligroin and treated with active carbon. The. hot filtrate was allowed to stand at room temperature to separate 2.9 g. of the crude product. Recrystallizations from ligroin gave almost pure 3~ (2, 4-dichlorophenyl) - -cyano~5-methyliso-thiazole as measured by GLC (SE-30, 230° C„, He 40 ml./min., retention time 3.9 min.). Yield 2.0 g., m.p. 107-110° C. 2260 cm"1. A„axH 8.5 mu (£ΐΟ, 4θθ) . NMR (100 Mc; in CCl^, ref. T S); 7 (ppm) 7.19 (s, -CH^), 2.57 (s, phenyl ring protons) .

Anal. Calc'd. for C^HgClgNgS: C, 49.08j H, 2.25; N, 10.41. Found; C, 49.29, ½8.79; H, 2.02, 1.98; N, 9 - 99, 9 - 99. 3- (2. 4-Dichlorophenyl - -methyllsothiazole-4-carboxylic acid (II) - A mixture of 1.66 g. (0.006 mole) of 3~ (2, 4-dichloro-phenyl) -4-cyano-5-methylisothiazole and 30 ml. of cone, sulfuric acid was heated on a water bath at 80° C. for 4 hours. The reaction mixture was cooled and a solution of 0.72 g. (0.104 at 0-5° C. After the addition was completed the mixture was stirred at room temperature for 0.5 hour, then at 70° C. for 0.5 hour and poured onto 100 g. of crushed ice. The separated product was filtered, washed with water and dissolved in 50 ml. of 2# aq. sodium bicarbonate solution, a small amount of in-soluble material being removed by filtration. The filtrate was acidified with 10$ hydrochloric acid to afford the product, 3-2, 4-dichlorophenyl-5~methylisothiazole-4-carboxylic acid, which was recrystallized from water-ethanol (is 2) . Yield 1.356 g. (7β ) , m.p. 178-179° C. 1. 7l?ma*x?H 255 mu (e8, 800) .

Anal. Calcd. f C, 45.85; H, 2.45; N, 4.86. Found: C, 45.52, 45.40; H, 2.55, 2.57; N, 4.91, 4.80. ^- (2.4-Dichlorophenyl) -5-methyllsothiazole-4-carbonyl chloride (III) - A mixture of 1. 1 g. (0.0038 mole) of 3- (2, 4-dlchlorophenyl) -5-methylisothiazole-4-carboxylic acid and 3 ml. of thionyl chloride was heated on a water bath at 80° C. for 0.5 hour. The excess thionyl chloride was distilled off under reduced pressure. The residual oil was dissolved in 10 ml. of dry benzene and treated with active carbon. The filtrate was evaporated under reduced pressure to give a viscous oil. The product, 3- (2, 4-dichlorophenyl) -5~methylisothiazole-4~carbonyl chloride, was used for the next reaction without further puri-fication. Yield 1.1 g. ( 94#) . 176° cm""1- Sodium 6-[ 3- (2.4-dichlorophenyl) -5-methyllsothiazole-4-carboxamldo]peniclllanate (IV) - To a stirred solution of 0.86 g. (0.004 mole) of 6-aminopenicillanic acid, 1.0 g. (0.012 mole) of sodium bicarbonate, 40 ml. of water and 25 ml. of acetone was added dropwise a solution of 1.1 g. (Ο.ΟΟ36 mole) of 3- (2, 4-dlchlorophenyl) -5-methylisothiazole-4-carbonyl chloride in 15 ml. 1 hour at 5° C., washed with two 100 ml. portions of ether, covered with 100 ml. of ethyl acetate and adjusted to pH 2.0 with 10$ hydrochloric acid at 5° C. The ethyl acetate layer was separated and the aqueous layer was extracted with two 40 ml. portions of ethyl acetate. The combined ethyl acetate extracts were washed with two 20 ml. portions of water and dried with anhydrous sodium sulfate. The filtrate was evapor-ated to 5 ml. below 40° C. under reduced pressure. The concen-trate was treated with 1.7 ml. of 3 $ SEH solution and then 100 ml. of n-hexane to give the product, sodium 6- [3- (2, -dichlorophenyl) -5-methylisothiazole-4-carboxamido ]penicillanate, which was collected by filtration and dried in vacuo over phos-phorous pentoxide. Yield 1.64 g. (8¾ , m.p. 170-175° C. (dec.) ^ 176 , 1640, 1600, 1400 cm"1. 258 ιημ (* 9, 4θθ) .

Anal. Calc'd for C^H^Cl^O^SgNa; C, 44.89J H, 3.17J N, 8.27. Pound: C, 46.14, 46.28; H, 4.06, 4.14; N, 7.23, 7.02.

This product in vitro exhibited Minimum Inhibitory Concentrations of about 0.4 meg./ml. versus S. aureus Smith and about 0.8 meg./ml. versus benzylpenicillin-resistant S. aureus Bx-1633-2 and in mice upon intramuscular injection exhibited of about 12 mgm./kg. versus S. aureus Smith and about 50 mgm./kg. versus S. aureus Bx-1633-2. This penicillin was also very stable in aqueous acid, having a half-life of about 4.2 hours at pH 2 and 37° C.

EXAMPLE 12 Sodium 6- (5-m-chlorophenyl-5-methylisothiazole-4-carboxamido)penlcillanate . - In similar fashion there was prepared 5-m-chlorophenyl-5-methylisothiazole-4-carboxylic acid (m.p. l47-l48° C.) which was converted to its acid chloride (b.p. 157-159° C./0.6 mm.) which was reacted with 6-aminopenicillanic acid to produce, sodium 6- (5-m-chloro-phenyl-5-methylisothiazole-4-carboxamido)penicillanate, melting at 215-218° C. with decomposition.

Anal. Calc'd for C^H^ClN^O^Na- 1 1/2 Η≥0 : C, 45.55 ; H, 4.25; N, 8.59. Pound: C, 45.57, 5.47; H, .5 , 4.22; N, 8.46, 8.66.

This product in vitro exhibited Minimum Inhibitory Concentrations of about 0.2-0.4 meg. /ml. vs. S. aureus Smith and about 0.8 meg./ml. vs. benzylpenicillin-resistant S. aureus Bx-1655-2 and was very stable in aqueous acid having a half-life of about 4.9 hours at pH 2 and 57° C.

EXAMPLE 13 N CHCOOK Potassium 6- (5-methyl-3-phenylisothiazole-4-carboxamido)penlclllanate . - A solution of 5-methyl-3-phenylisothiazole-4-carboxylic acid (O.Ol mole; prepared as shovm in Example l) in a mixture of 20 ml. of dimethylformamide and 35 ml. of methylene chloride is cooled to -5° C. To this solution is added ethyl chloroformate (0.01 mole) which causes the temperature to rise to -1° C. The reaction mixture is cooled and stirred for 0 minutes at -5° C. A solution at -2° C. of 6-aminopenicillanic acid ( 0.01 mole) in 25 ml. of methylene chloride and triethylamine ( 0.02 mole) is added in one portion to the first reaction mixture. The temperature rises to 0° C. and carbon dioxide is evolved for several minutes. The reaction mixture is stirred for 40 minutes at -5° C. The methylene chloride is removed by distillation under reduced pressure at 35° C. and the resulting mixture is diluted to 200 ml. with dry ether and then filtered. Potassium 2-ethyl-hexanoate ( 2 gm.) dissolved in a small amount of ethyl acetate is added to the filtrate whereupon a white crystalline precipitate is formed. The precipitate is collected by filtration and is dissolved in 50 ml. of water, acidified to pH 1.8 with sulfuric acid. The aqueous solution is extracted with 2 ml. portions of ether. The aqueous solution is then adjusted to four 2Q ml. portions of cold ethyl acetate. S e ethyl acetate extract is dried over anhydrous sodium sulfate and potassium 2-ethylhexanoate is added to precipitate the product. The product, the potassium salt of 6-(5-methyl-3-ph.enylisoth.ia- zole-4-earboxamldo)penicillanic acid is collected by filtration, dried over ?2°5» and found ¾o ®®1* a* 180-190° C. with decomposition. aiARjFLE 13 (a) 3- (2-chloro-6-fluorophenyl)-5-methyl 4-isoxa-501e- earboxylie acid.

To a stirred and caied slurry of 52 g, (0.3 mole) of 2-chloro-6-fluorobengaldoxime S.A. 31: 381δ ; Ber. 69B, 2253-8 in 300 ml. of chloroform at -10°C. was added 24 g. (0,33 mole) of chlorine in 300 ml. of chloroform over a twenty minute period. The cooling bath was then removed and stirring continued for 2.5 hours. She CHCl^ was then removed at 20°C. under reduced pressure and the residual oil dissolved in 100 ml, of methanol and added over a one minute period to a -30°C. stirred solution of 300 ml. methanol, 1β.*2 g, (0.3 mole) of sodium methoxide and 39 g. (0.3 mole) of ethyl acetoacetate. The exothermic reaction was controlled with a Dry Ice- acetone bath to keep the temperature below -10°C. After 30 minutes at 0°C. to -5°C, the bath was removed and stirring continued for 12 hours at room temperature (22°C). The ° shaken with 300 ml. other and 300 ml. of water. The ether layer was washed with three 100 ml. portions of water and then evaporated to an oil. The oil was then saponified at reflux for 2 hours in a solution of 300 ml. methanol, 100 ml. HgO and 24 g. of sodium hydroxide. The methanolic solution was then concentrated to renove the methanol and 300 ml. of water added. The aqueous solution was extracted with two 300 ml. portions of sthor and then placed on the rotary flash evaporator to remove dissolved ether. The solution was then cooled and stirred while being acidified to pH 2 with Q# phosphoric acid. The crystalline product was then filtered off» washed several times with water and air dried. One recrystallization from methanol- ater gave 52 g.i M.F . 205-20S°0.

IE and HMR spectra were consistent with the desired structure.

Analysis: Calc'd for CnH?CMi( C, 51.8» H, 2.76j H, 5.49i Pound Ϊ C, 51.9» H, 2.75j ff, 5.67. (b) Methyl 3-(2-chloro-6-fluorophenyl)-5-methyl-4-isoxazole-carboxylate.

To 51 g. (0.2 mole) of 3-(2-chloro-6-fluorophenyl)-5-net¾yl-4-isoxazolecorboxylic acid was added 100 ml. of thionyl chloride and the mixture heated at gentle reflux for four hours. The excess SOClg was removed at 25°C under reduced pressure and the crude acid chloride dissolved in 300 ml. of methanol. This solution was rofluxed for three hours and then concentrated to an oil under reduced pressure. She residue was crystallized in "Skellysolve B" (petroleum ether) and recrystallized from cyclohexane to give 37 g.» K.P. 55-56°C.

The IR and ΜΊ& spectra were entirely consistent with the Analysis t Calc'd for C, 53.53» H, 3.34; H, 5.19 Found ί 0, 53.69j H* 3.45 i M, 4,96. (c) l-ami3o-l-(2-.c loro-6-fluorop enyl)-2-carbomethoxy-l-buten-3-one. ϊο 27 g. (0.1 mole) of methyl 3*<2-chloro-S-fluorophenyl)-5~aethyl-4-lsoxaizolecarboxylate in 150 ml. of methanol was added approximately 5 g. of commercial Eaney nickel catalyst and the mixture hydrogenated at an initial pressure of 50 p.s.i. for 5.5 hours, at which time the Parr apparatus showed an uptake of 9 lbs. (0.1 mole) of Hg. The catalyst was then filtered off, washed with methanol and the combined filtrates concentrated to an oil under reduced pressure at 20°C. The oil slowly crystallized but a suitable recrystalliza ion solvent was not found due to its very low melting point. The product was used "as lsw for the next step. (dj 3- (2-chloro-6-fluorophenyl)-5-methyl-4-isothiazole-carboxylic acid.

A mixture of 27.1 g. (o.l mole) of l-amino-l~(2-ehloro-6-fluorophenyl)-2-carbomethoxy-l-buten-3-one, 66.6 g. (0.3 mole) of 24.6 g. (©.1 mole) of chloranil in 600 ml. of toluene was stirred at reflux for 20 minutes, The mixture was filtered and the filtrate evaporated under reduced preeeure at _0°C. to an oil. To this oil was added 900 ml. of benzene and the solution filtered to remove a small amount of solids, fhe solution was concentrated under reduced pressure at 20°C. to a volume of about 100 ml. This solution was placed on a 26 x 7i cm. column of Merck alumina and eluted with 3.5 liters of benzene. Concentration of the eluate under reduced pressure left an oil whieh was sa onified at reflux in 150 ml. methanol IE and 1HR spectra were entirely consistent with the . desired structure.

Analysis.

Calc'd for CuH7<31FIG2Si C, 48*531 H, 2.59» S, 11.76 Found t C, 49.3Q> 48.96} Ht 2.89, 2.71} S, 11.80, 11.88. (e) Sodium 6- -(2-chloro-6-fluorop enyl)-5-methy1-4-iso-thia^olecarboxaaido/.penicilianate monohydrate.

To 6.8 Q, (0.025 mole) of 3-{2-ehloro*6-fluorophenyl)-5-methyl-4-isothia¾olecarboxyllc acid w s added 30 ml. of thionyl chloride and the mixture heated at reflux for 1.5 hour© on the steam bath. The excess of SOCl^ was then removed at 20° C. under reduced pressure. The remaining oil was dissolved in 50 nl. of acetone and added, all at once,t© a vigorously stirred solution of 5.4 g. (0.025 mole) of 6-ΔΡΔ, 8.4 g. (o.l mole) of M&HCG^ la 100 ml. of water and 50 ml. of acetone at 5°C. After 10 minutes the ice bath was removed and stirring was continued for one and one half hours. The acetone s then removed under reduced pressure at 20°C. Fifty ml. of water was added and the solution extracted with wo 100 ml. portions of ether and the ether extracts discarded. The aqueous phase wao then layered with 75 ml. of methyl isobutyl ketone (KSIBK) portions of water and two times with 75 ml. portions of saturated Na^SO^ solution. The MIBK solution was then filtered through anhydrous M&^SQ^ and the HSgSO^ cake washed with §0 ml. of MIBK and the filtrates combined. The MIBK solution was then treated with 8 ml. of sodium 2-ethylhexanoate in n-butanol (concentration of 34 ml. « 0.1 mole). Scratching induced crystallization and 50 minutes later the product was filtered off, washed with three 100 ml. portions of acetone and air dried. After drying 24 hours under vacuum over there was 8.5 g. of snow white crystals with a decomposition point of 175-176°C.

The IK and MR spectra were entirely consistent with the desired structure.

Analysis! o Calc'd for C^H^Cl i^O^Ias C, 46.34} H» 3.2 s I» 8.54 Pound ί C, 46.25* H, 3.44* H, 8.20.

(Corrected for 2.67$ H20 determined by the Karl Fischer method,). tthil© in the foregoing specification various embodiments of this invention have been set forth in specific detail and elaborated for the purpose of illustration, it will bo apparent to those skilled in the art that this invention is susceptible to other embodiments and that many of the details can be varied widely without departing from the basic concept and the spirit and scope of the invention.

Claims (3)

1. -52- 25756/2 C B3S 1. A compound of the formula » In which is -QMt -C-M2, -C-QB or Its reactive functional derivatives as heroin defined, or the group R is hydrogen, chloro or trifluoroaethyl, H is hydrogen, c loro, trifluoromethyl, fluoro, nitro, lovier alkyl or lower alkoxy, and S Is lower alkyl; and non-toxic pharmaceu ically acceptable salts of those compounds In which 2 is the group (A) .

2. Compounds of the formula u erein R"*", R and R have the same r.eaning as in Claim 1; and nontoxic, pharmaceutically acceptable ealts thereof. 5. A compound having the formula wherein R represents (lower )alk 1; and nontoxic pharmaceutically acceptable salts thereof. - 53 - 4 A compound having the formula wherein R and R each represent (lower Jalkyl; and nontoxic, pharmaceutically acceptable salts thereof. A compound having the formula wherein R represents (lower Jalkyl; and nontoxic, pharmaceutically acceptable salts thereof. Α compound having the formula wherein R represents (lower )alkyl, and R1 represents (lower)-alkoxy; and nontoxic, pharmaceutically acceptable salts thereof. CI wherein R represents (lower )alkyl; and nontoxic, pharmaceutically acceptable salts thereof. A compound having the formula whereln R represents (lower Jalkylj and nontoxic, pharmaceutically acceptable salts thereof. wherein R represents ( lower)alkyl; and nontoxic, pharmaceutically acceptable salts thereof. wherein R represents (lower )alkyl; and nontoxic, pharmaceutically acceptable salts thereof. and nontoxic, pharmaceutically acceptable salts thereof. The compound having the formula and nontoxic, pharmaceutically acceptable salts thereof. and nontoxic, pharmaceutically acceptable salts thereof. The compound having the formula and nontoxic, pharmaceutically acceptable salts thereof and nontoxic, pharmaceutically acceptable salts thereof and nontoxic, pharmaceutically acceptable salts thereof. and nontoxic, pharmaceutically acceptable salts thereof. «61- 25756/2 -v 18* A compound according to Claim 1, of the general formula: , wherein 2· and R 3 have the same meaning as in Claim 1. 19· conpound according to Claims 1 and IS, having the general formula* it n 1 2 wherein 2' represents -CII, -C-iffig or -C-OH, and E and R each represent hydrogen or chloro. 20· A compound according to Claims 1, 18 and 19» having the general formulas wherein Z* represents —-C0M1, —- Cc^—- fI™Hg or -C-O rttHr, and a · Rη a „nd I S «2 each represent hydrogen or chloro. -62 23756/2 * 21. A process for the preparation of compounds of fornula I in Claim 1 in which 2 is the group (A), and their salts, which comprises reacting 6-aminopenicillanie acid or a sal thereof with at least one equivalent of a reactive functional derivative (as herein defined) of the corresponding acid of formula 2 in Claim 1 in which Z is -COOH. 22. L process according to Claim 21, wherein the acid chloride is used as the reactive functional derivative. 2

3. A process according to Claim 21, wherein a mixed acid anhydride is used as the reactive functional derivative. 24· process according to Claim 21 or 23, wherein in the sdxed acid anhydride the second acid moiety is derived from a lower alfcyl raonoester of carbonic acid. 25. A process for the preparation of compounds of formula I in Claim 1 in which 2 is the group (A), and their salts, which comprises reacting β-aminopenicillanie acid or a salt thereof with at least one equivalent of the corresponding acid of ormula I in Claim 1 in which Z is -COOH in the presence of a earbodiimide reagent. 26. A process according to any one of Claims 21 to 25, wherein the reaction is conducted at a temperature of from about -50°C to about +50°C, preferably f o® about -5°C to about *15°0. 27· A process for the preparation of compounds of formula Ϊ in Claim 1 substantially as hereinbefore described with reference to the Examples. For the Ap licants