US 4,033,950 AGrant
3-Hydroxymethyl-7.beta.-(2-Alkoxy-or Benzyloxyimino-2-Arylacetamido)ceph-3-Em-4-Carboxylic Acids and Salts Thereof
Issue Date:1977-07-05
•3 Claims
Abstract
The invention provides novel antibiotic compounds which are 7.beta.-acylamidoceph-3-em-4-carboxylic acids, and non-toxic derivatives thereof characterized in that the acylamido group has the structure ##STR1## where R is a hydrogen atom or an organic group and R.sup.a is an etherifying monovalent organic group linked to the oxygen atom through a carbon atom. The compounds are syn isomers or exist as mixtures containing at least 75% of the syn isomer. These antibiotic compounds possess high antibacterial activity against a range of gram positive and gram negative organisms coupled with particularly high stability to .beta.-lactamases produced by various gram negative organisms. The invention is also concerned with the administration of the compounds.
Metadata
Assignee
- Glaxo Laboratories Limited
Inventors
- Martin Christopher Cook
- Gordon Ian Gregory
- Janice Bradshaw
Application Information
Application Number:US 6698182
Filing Date:1976-03-24
Priority Date:1971-05-14
Art Unit:122
Classifications
IPC:
C07D50132C07D50134
Field of Search:
260243 C
Patent Drawings
This patent does not have any drawings.
Description
The following examples illustrate the invention.
PREPARATION 1
2-Methoxyiminophenylacetic acids (syn- and anti-isomers).
A solution of sodium (5 g.) in dry methanol (100 ml) was added to a solution of O-methylhydroxyamine hydrochloride (15 g.) in dry methanol (100 ml) until neutral to phenolphthalein. The precipitated sodium chloride was removed by filtration, and the filtrate added to a solution of phenylglyoxylic acid (25 g.) in dry methanol (100 ml). The solution was refluxed for 2 hours, cooled, and evaporated to an oil, which was dissolved in ether (200 ml), refiltered and evaporated to an oil (32.9 g.). This was crystallised from petroleum spirit, (bp. 60.degree.-80.degree. ) producing a white solid (19.61 g.) and oil (3.9 g.).
The solid (17.8 g.) and the oil (3.9 g.) were combined (21.7 g.) and methylated with ethereal diazomethane, producing an oil (24.2 g.). This was purified by chromatography on silica gel (600 g.), producing syn-methyl 2-methoxyiminophenylacetate as an oil (13.6 g. 55%), .lambda..sub.max. (EtOH) 259 nm (.epsilon. 10,400) and anti-methyl 2-methoxyiminophenylacetate, the slower component, as a solid (8.7 g.; 35%), m.p. 54.degree., .lambda..sub.max. (EtOH) 251 nm (.epsilon. 7,260).
Methyl 2-methoxyiminophenylacetate (anti-isomer) (8.7 g.) was dissolved in methanol (100 ml) and 2N-sodium hydroxide solution (22 ml.) was added. The solution was stirred at room temperature for 1 hour, and the pH adjusted to 7 with 2 N-hydrochloric acid. Methanol was removed by evaporation, water (150 ml.) was added, and the solution acidified to pH 1.5 with 2 N-hydrochloric acid. The mixture was extracted with ethyl acetate (3.times. 100 ml), the organic extracts were combined, dried and evaporated to give a solid (6.74 g.) which was crystallised from petroleum spirit (b.pt. 60.degree.-80.degree. ): benzene, producing anti-2-methoxyiminophenylacetic acid (4.84 g.), m.p. 103.degree.-104.degree., .lambda..sub.max. (EtOH) 248 nm (.epsilon. 7,010). .tau. (CDCl.sub.3) values include 2.64 (Ph), 5.92 (CH.sub.3).
Methyl 2-methoxyiminophenylacetate (syn-isomer) (13.6 g.) was hydrolysed in a similar manner, but the hydrolysis mixture was stirred for 40 hours at room temperature. The white solid (11.13 g.) formed was crystallised from petroleum spirit (b.pt. 60.degree.-80.degree. ); benzene producing syn-2-methoxyiminophenylacetic acid as a white solid (10.02 g.). m.p. 96.degree.-97.degree., .lambda..sub.max. (EtOH) 255 nm, (.epsilon. 13,200), .tau. (CDCl.sub.3) values include 2.2-2.8 (Ph), 5.92 (CH.sub.3).
PREPARATION 2
2-Methoxyimino-(thien-2-yl)-acetic acids (syn- and anti-isomers)
A solution of methoxyamine hydrochloride (5.85 g.) in dry methanol (60 ml.) was neutralised (phenol-phthalein) with a solution of sodium methoxide in methanol [from sodium (2.5 g.) and dry methanol (50 ml.)]. The precipitated sodium chloride was removed by filtration, and the filtrate was added to a solution of thien-2-ylglyoxylic acid (10 g.) in dry methanol (60 ml.). The resulting solution was refluxed for 1 hour, cooled, and evaporated to an oil. Ether (100 ml.) was added, the mixture was filtered, and the filtrate was evaporated to an oil (13.06 g.).
The oil (12.5 g.) was dissolved in ether (50 ml.) and an ethereal solution of diazomethane was added until a permanent yellow colour remained. The excess diazomethane was destroyed by leaving the solution in sunlight for 1 hour. Evaporation of this solution produced an oil (13.2 g.).
The oil (10.33 g.) was purified by preparative plate chromatography (Kieselgel PF.sub.254+ 366) developing three times with 75% petroleum spirit (b.p. 60.degree.-80.degree.) in benzene, producing
(a) methyl 2-methoxyimino-2-(thien-2-yl)-acetate (syn isomer) (3.44 g., 27%), .lambda..sub.max..sup.EtOH 290 nm (.epsilon. 11,250), .lambda..sub.inf. 271 nm (.epsilon.5,400) .nu..sub.max. (CHBr.sub.3) 1738 and 1230 cm.sup.-.sup.1 (CO.sub.2 Me). .tau. values (CDCl.sub.3) include 6.06 (s, CO.sub.2 CH.sub.3), 5.78 (s, OCH.sub.3).
(b) methyl 2-methoxyimino-2-(thien-2-yl)-acetate (anti-isomer) (1.21 g., 9.5%), .lambda..sub.max..sup.EtOH 221 and 288 nm (.epsilon. 5,020 and 11,000). .nu..sub.max. (CHBr.sub.3) 1732 and 1212 cm.sup.-.sup.1 (CO.sub.2 Me), .tau. (CDCl.sub.3) values include 6.06 (s, CO.sub.2 Me), 6.00 (s, OCH.sub.3) and further fractions which were isomeric mixtures.
2N-Sodium hydroxide (8.27 ml.) was added to a solution of methyl 2-methoxyimino-2-(thien-2-yl)-acetate (syn-isomer) (3.28 g.) in methanol (50 ml.) and the solution was stirred at room temperature for 18 hours. Water (20 ml.) was added and the solution was evaporated to remove methanol, and then washed with ethyl acetate. The pH of the solution under ethyl acetate (50 ml.) was altered to 2 with 2 N-hydrochloric acid. The layers were separated and the aqueous phase was extracted with ethyl acetate. The organic extracts were combined, dried, and evaporated to a white solid (2.58 g.). This was crystallised from cyclohexane, producing the title compound (syn-isomer) (2.23 g., 73%), m.p. 105.5.degree., .lambda..sub.max..sup.EtOH 289 nm (.epsilon. 10,100), .lambda..sub.inf. 262 and 271 nm. (.epsilon. 7,750 and 8,150), .tau. (CDCl.sub.3) values include 0.32 (OH) and 5.92 (OCH.sub.3).
Similar hydrolysis of the anti-methyl ester gave 2-methoxyimino-(thien-2-yl)-acetic acid (anti-isomer) (0.85 g.), .lambda..sub.max..sup.EtOH 286-287 nm (.epsilon. 10,200), .tau. (CDCl.sub.3) values include 1.31 (OH) and 5.73 (OCH.sub.3).
PREPARATION 3
syn-t-Butoxyiminothien-2-ylacetic acid
A solution of thien-2-ylglyoxylic acid (6.2 g.) and sodium bicarbonate (3.36 g.) in water (100 ml.) was added dropwise to a stirred solution of t-butoxyamine hydrochloride (5.65 g.) and sodium bicarbonate (3.78 g.) in water (100 ml.) at 0.degree.-5.degree. and the mixture was stirred at room temperature for 18 hr. The mixture was acidified with 2 N hydrochloric acid to pH 2.0 and extracted with ethyl acetate. The combined extracts were washed with water, dried and concentrated to give a solid (9.75 g.). Recrystallisation from petroleum (b.p. 60.degree.-80.degree.) gave the title compound (4.0 g., 44%), m.p. 106.degree.-107.degree., .lambda..sub.max. (EtOH)290 nm (.epsilon. 11,600), .tau. (CDCl.sub.3) values include 2.46, 2.66, 2.98 (d doublets, thienyl protons), 8.60 (C(CH.sub.3).sub.3).
PREPARATION 4
2-Ethoxyiminophenylacetic acids (syn and anti-isomers)
Ethoxamine hydrochloride (4.0 g.) and phenylglyoxylic acid (6.0 g.) were dissolved in water (50 ml.), and the resulting solution was basified to pH 4.5, and stirred at this pH for 15 hr. Acidification and extraction of the mixture gave, after evaporation of the ethyl acetate, a mixture of syn and anti ethoxyiminophenylacetic acids (7.4 g., 94%). Four recrystallisations from cyclohexane failed to give the pure syn acid. However, evaporation of the mother-liquors from the first crystallisation, and recrystallisation of the residue from cyclohexane gave anti-ethoxyiminophenylacetic acid (1.36 g., 17%), m.p. 90.9.degree.-91.6.degree., .lambda..sub.max. (ethanol) 249 nm (.epsilon. 7,600), .tau. (DMSO-d.sub.6) values include 2.52 (s, Ph) 5.74 (q, CH.sub.2), 8.76 (t, --CH.sub.3).
A solution of the mixed acids (4.0 g.) in ether (100 ml.) was treated with an ethereal solution of diazomethane until a yellow colour persisted. Acetic acid was added to destroy excess diazomethane and the ether solution was washed with sodium bicarbonate solution, water, and brine, then dried. Evaporation of the ether gave the methyl esters (4.1 g.) as an orange oil. These were separated on five 40.times. 20 cm. preprative plates, eluting with petroleum spirit (b.p. 40.degree.-60.degree.)/ether (3:1). The slower band was eluted with chloroform, and removal of the solvent gave anti-methyl 2-ethoxyiminophenylacetate (1.45 g.) as a pale-yellow oil; .tau. (CDCl.sub.3) values include 2.58 (Ph), 5.66 (q, CH.sub.2), 6.12 (s, OCH.sub.3), 8.72 (t, CH.sub.3). Similar treatment of the faster band gave syn-methyl 2-ethoxyiminophenylacetate (2.45 g.) as a pale yellow oil, .tau. (CDCl.sub.3) values include 2.3-2.7 (m, Ph) 5.72 (q, CH.sub.2), 6.06 (s, OCH.sub.3), 8.67 (t, CH.sub.3).
The above syn-methyl ester (2.39 g.) in methanol (60 ml.) was treated with sodium hydroxide solution (2N; 12 ml.), and the solution was stirred for 18 hr. The methanol was removed, and the aqueous mixture, after being acidified to pH 1.5, was extracted with ethyl acetate. The washed and dried extracts were evaporated to dryness, and the residue was recrystallised from cyclohexane to give syn-ethoxyiminophenylacetic acid (836 mg.), m.p. 77.9.degree.-79.0.degree., .lambda..sub.max. (ethanol) 256.5 nm (.epsilon. 12,800); .tau. (DMSO-d.sub.6) values include 2.48 (m, Ph), 5.74 (q, CH.sub.2), 8.71 (t, CH.sub.3).
PREPARATIONS 5-23
2-Alkoxyiminoarylacetic Acids
General Procedures
A mixture of the substituted glyoxylic acid and an excess (10 to 15%) of the alkoxyamine hydrochloride was suspended in water or aqueous ethanol, stirred, and the pH of the mixture adjusted to between 4 and 5 (Method B) with sodium hydroxide solution (N to 10 N). A clear solution at pH 4 to 5 was maintained during the reaction by further additions of sodium hydroxide solution and ethanol as needed. The reaction mixture was kept at room temperature until all of the keto-acid was consumed (it may be necessary to add a further portion of the more volatile alkoxyamines). The progress of the reaction was followed by acidification of an aliquot, extraction with ethyl acetate and thin layer chromatography of the extract on silica plates (developed with a mixture of chloroform; methanol: acetic acid; 18:2:1). The alkoxyiminoacetic acids were less polar than the starting keto-acids. The reaction times were 2 hr. to 2 days. Reactions carried out at pH 7-8 are designated Method A. When reaction was complete the pH of the mixture was adjusted to between 7 and 8 and the ethanol (if any) was removed by evaporation. The aqueous mixture was extracted with ether, the extract discarded and the aqueous phase acidified to pH< 2 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate, the extract dried and evaporated to give the crude product which was purified by one of the following methods:
(a) Crystallisation and recrystallisation (if needed) from a suitable solvent,
(b) The crude product dissolved in ether was treated with a small excess of a solution of diazomethane in ether. The excess reagent was destroyed with acetic acid and the solution washed with sodium bicarbonate solution and evaporated to give the crude methyl esters. The esters were separated by preparative thick layer chromatography or column chromatography on silica, and then hydrolysed conventionally with alkali to give the pure syn acid,
(c) The mixture of methyl esters was prepared as in (b) and the isomers separated by crystallisation from a suitable solvent and similarly hydrolysed.
These methods were employed to prepare the intermediates listed in Table 1 (syn-isomers)
PREPARATION 24
2-Methoxyiminopyrid-3'-ylacetic acid (syn- and anti-isomers)
Pyrid-3-ylglyoxylic acid (3.02 g.) and methoxylamine hydrochloride (2.59 g) were heated under reflux in ethanol (50 ml.) and treated with a solution of sodium ethoxide in ethanol until the mixture was just alkaline to phenophthalein. The mixture was maintained under reflux for 11/2 hours. Further portions of methoxylamine hydrochloride (0.5 g) and ethanolic sodium ethoxide were added and refluxing continued for a further 1 hour. The resulting mixture was filtered through Kieselguhr and evaporated to dryness. The residue was dissolved in a small volume of ethanol and the remaining fine suspension was removed by filtration through Kieselguhr. Dilution of the filtrate with ethyl acetate gave a crystalline solid. The solid was washed well with ethyl acetate and dried to yield 2-methoxyiminopyrid-3'-ylacetic acid as an isomeric mixture (2.72 g., 76%), .lambda..sub.max. (pH 6 buffer) 250 nm (.epsilon. 6,630), .nu..sub.max. (Nujol) 1610 (CO.sub.2 .sup.-) and 1038 cm..sup.-.sup.1 (=C--O--), .tau. (d.sub.6 DMSO) 1.21, 1.43, 2.02, 2.60 (multiplets; aromatic protons), 6.20 (s; OCH.sub.3).
PREPARATION 25
(a) Methyl 2-(1-ethoxy)ethoxyimino-2-(thien-2-yl)acetate (syn-isomer)
To a stirred mixture of methyl 2-hydroxyimino-2-(thien-2-yl)acetate syn-isomer (3.98 g.) and ethyl vinyl ether (2.5 mls) in ethyl acetate (25 mls) was added phosphorous oxychloride (2 drops). After 20 mins. at 50.degree. the ethyl acetate was washed with saturated sodium bicarbonate solution, dried over sodium sulphate and evaporated to an oil, giving methyl 2-(1-ethoxy)ethoxyimino-2-(thien-2-yl)acetate (syn-isomer) (5.7 g; 100%) .lambda..sub.max. (EtOH) 289 nm (.epsilon. 11,700), .tau. (CDCl.sub.3 ; 60 MHz) 2.61 (multiplet; thienyl H.sub.5), 2.82 to 2.97 (multiplet; thienyl H.sub.3 and H.sub.4), 4.64 (quartet, J5 Hz; ##STR57## 6.06 (singlet, --COOCH.sub.3), 6.24 (quartet, J 7 Hz, OCH.sub.2), 8.56 (doublet J 5 Hz; CH--CH.sub.3), 8.79 (triplet, J 7 Hz; O.CH.sub.2 CH.sub.3).
(b) 2-(1-Ethoxy)ethoxyimino-2-(thien-2-yl)acetic acid sodium salt (syn-isomer)
1 N-Sodium hydroxide (1 equiv.) and enough methanol to form a homogeneous system were added to methyl 2-(1-ethoxy)ethoxyimino-2-(thien-2-yl)acetate (syn-isomer) (5.7 g.). After 4 hrs. at 50.degree. the methanol was evaporated and the residue azeotroped with benzene/methanol giving a white solid, 2-(1-ethoxy)ethoxyimino-2-(thien-2-yl)acetic acid sodium salt syn-isomer) (4.6 g, 78.5%), .lambda..sub.max. (pH 6 buffer) 287.5 nm (.epsilon. 10,650), .tau. (D.sub.2 O) values include 2.42 (multiplet; thienyl H.sub.5), 2.68 to 2.84 (multiplet; thienyl H.sub.3 and H.sub.4), 4.63 (quartet, J 5 Hz; ##STR58## 6.21 (quartet, J 7 Hz; --CH.sub.2 --CH.sub.3), 8.57 (doublet, J 5 Hz; ##STR59## 8.82 (triplet, J 7 Hz; --CH.sub.2 --CH.sub.3).
PREPARATION 26
syn-Then-2-yloxyimino(thien-2-yl)acetic acid
Then-2-yloxamine hydrochloride (7.37 g.) and thien-2-ylglyoxylic acid (6.24 g.) were dissolved in ethanol (110 ml.) and water (20 ml.). The pH of the solution was adjusted to 5.0, and this solution was stirred for 22 hr. The ethanol was evaporated off, and the aqueous mixture was neutralised, washed twice with ether, then acidified to pH 1.5. The acid mixture was extracted with ethyl acetate. The combined extracts were washed, dried and evaporated to dryness to give a yellow oil (9.2 g., 86%) which crystallised on standing. TLC indicated the solid to be a mixture of isomers. Recrystallisation of this solid several times from cyclohexane effected no separation of isomers. The mixture of acids (5.0 g.) was esterified with diazomethane to give the mixture of methyl esters, as a pale-yellow oil.
To a solution of the mixture of methyl esters (2.14 g.) in methanol (50 ml.) was added 2 N sodium hydroxide solution (7.6 ml.). After stirring this solution for 0.5 hr., the solution was neutralised. The methanol was evaporated and the aqueous residue was extracted with ethyl acetate. The extracts were washed, dried, and evaporated to dryness to give a yellow oil (1.0 g.). This oil was dissolved in methanol (25 ml.) and stirred for 18 hr. with 2 N sodium hydroxide solution (5 ml.). Methanol was removed by evaporation and the aqueous residue, after washing with ethyl acetate then acidification to pH 1.7, was extracted with ethyl acetate. The extracts were washed, dried, and evaporated to dryness to give a solid (730 mg.). Recrystallisation of this solid twice from cyclohexane gave syn-then-2-yloxyimino(thien-2-yl)acetic acid (369 mg.), m.p. 101.degree.-102.degree., .lambda..sub.max. (EtOH) 239, 289.5 nm (.epsilon. 11,700, 12,300), .tau. (DMSO-d.sub.6) values include 4.67 (s, CH.sub.2).
PREPARATION 27
syn-2-Benzyloxyiminobenzo[b]-thien-3'-ylacetic acid
Benzo[b]-thien-3-ylglyoxylic acid (2.27 g) and benzyloxyamine hydrochloride (1.915 g) were dissolved in ethanol (70 ml) and water (30 ml). The solution was adjusted to pH 4.5 with 40% w/v sodium hydroxide solution and stirred at this pH for 2 hr. The solution was stood overnight and adjusted to pH 9 then washed with ether. The aqueous phase was acidified under ethyl acetate and the organic layer was washed with water, saturated brine and dried. Evaporation gave a mixture of syn and anti-isomers as a buff crystalline solid (3.4 g., 99%). The crude acid in ether was treated with excess diazomethane in ether at 0.degree.-5.degree.. The excess reagent was destroyed with acetic acid and the ether solution was washed with sodium bicarbonate, water and dried. Evaporation gave a pale brown oil (3.34 g., 93%). The crude product in methanol (100 ml.) was treated with sodium hydroxide solution (1 N, 10 ml) at room temperature for 1 hr. The hydrolysis was followed by thin layer chromatography on silica. Hydrochloric acid (2 N 5 ml.) was added to stop the hydrolysis and methanol was removed by evaporation. Ethyl acetate was added and the anti-2-benzyloxyimino benzo[b]-thien-3'-ylacetic acid was removed by washing with sodium bicarbonate. The ethyl acetate layer was washed with water and dried and evaporated to a pale orange oil (1.99 g., 56%). This was treated in methanol (90 ml.) with sodium hydroxide (1 N; 10 ml) at room temperature for 7 hr. A further aliquot of sodium hydroxide (1 N; 5 ml) was added and the solution stood for 2 days to complete hydrolysis. The methanol was removed by evaporation and the residue dissolved in ethyl acetate and water. The mixture was adjusted to pH 1.5 and the ethyl acetate layer was washed with water, saturated brine and dried. Evaporation gave yellow crystals (1.82 g, 50%). Crystallisation from a mixture of benzene and cyclohexane gave the title compound as pale orange crystals (1.29 g., 36%), m.p. 120.5.degree.-121.degree., .lambda..sub.max. (EtOH) 232, 285.5, 296.5 306.5 nm. (.epsilon. 22,500, 11,800, 11,500, 10,400), .tau. values (DMSO-d.sub.6) include 1.90, 1.97, 2.3-2.7 (aromatic protons), 4.64 (CH.sub.2 singlet).
PREPARATION 28
syn-2-Benzyloxyiminobenzo-[b]-thien-2'-ylacetic acid
Benzo[b]-thien-2-ylglyoxylic acid (3.092 g) and benzyloxyamine hydrochloride (2.72 g.) in ethanol (170 ml.) and water (70 ml.) were adjusted to pH 4.5 with sodium hydroxide (40%). The solution was stirred at this pH at room temperature for 6 hr. Benzyloxyamine hydrochloride (500 mg.,) was added and the solution stood at room temperature overnight. The solution was adjusted to pH 8 and washed with ether. The aqueous phase was acidified under ether to pH 1.5. The ether layer was washed with water and dried. Evaporation gave a cream coloured solid (4.28 g, 91%) as an isomeric mixture.
The crude isomeric mixture was treated in ether with excess diazomethane in ether at 0.degree.-5.degree.. The excess reagent was destroyed with acetic acid and the ether solution was washed with sodium bicarbonate, water and dried. Evaporation gave an oil (4.45 g., 91%). This was dissolved in methanol (140 ml.) and treated at room temperature with sodium hydroxide solution (1 N; 14 ml.) for 21/4 hr. Hydrochloric acid (2 N, 7 ml.) was added and the alcohol was removed by evaporation. The aqueous phase was partitioned between sodium bicarbonate solution and ether. The ether layer was washed with water and dried. Evaporation gave an oil (2.16 g., 44%). This was hydrolysed directly in refluxing methanol (70 ml.) with sodium hydroxide (1 N; 7 ml) for 4 hr. The methanol was removed by evaporation and the residue partitioned between water and a little ether. The aqueous layer was acidified under ether to pH 1.5 and the ether layer was washed with water, dried and evaporated to give a pale cream solid (1.97 g, 42%). Crystallisation from a mixture of benzene and cyclohexane gave the title compound as a white crystalline solid, (1.61 g; 35%), m.p. 141.degree.-143.degree. (dec.),.lambda..sub.max. (EtOH) 230.5, 253, 297.5 nm (.epsilon. 16,400; 7,400; 24,100), .tau. (DMSO-d.sub.6) values include 2.00, 2.36, 2.55 (aromatic protons), 4.71 (CH.sub.2 singlet).
PREPARATION 29
(a) (2-t-Butoxycarboxamido)ethoxyimino-thien-2-ylacetic acid (syn-isomer)
N-Carbo-t-butoxy-2-bromoethylamine (1.12 g.) was added to a solution of the sodium salt of methyl syn-hydroxyimino-thien-2-ylacetate (1.035 g.) in benzene: dimethylformamide (2:1 v/v, 30 ml.), and the mixture was stirred for 16 hr. Ethyl acetate (50 ml.) was added, and the mixture was washed several times with water, dried, and evaporated to dryness to give methyl syn-(2-t-butoxycarboxamidoethoxy)imino-thien-2-ylacetate (1.21 g, 75%), .tau. (CDCl.sub.3) values include 2.62, 2.86, 2.99 (thienyl protons), 5.10 (NH), 6.06 (s, CH.sub.3), 8.58 (s,C(CH.sub.3).sub.3).
The crude ester (1.1 g) in methanol (20 ml) was treated with 2 N sodium hydroxide solution (3.4 ml.), and stood 16 hr. The methanol was evaporated off, and the aqueous residue, after washing with ether, was acidified to pH 2.0, and extracted with ethyl acetate. The extracts were washed (water, saturated brine), dried, and evaporated to dryness. Recrystallisation of the residue from cyclohexane gave the title compound (951 mg., 90%), m.p. 112.8.degree.-114.4.degree. .lambda..sub.max. (EtOH) 290.5 nm. (.epsilon. 11,600), .tau. (DMSO-d.sub.6) values include 2.19, 2.6-2.9 (thienyl protons), 3.14 (NH), 8.52 (s, C(CH.sub.3).sub.3).
The alkylating agent used for the above preparation was made as follows:
(b) N-Carbo-t-butoxy-2-bromoethylamine
A mixture of t-butyl azidoformate (15.81 g) and triethylamine (30 ml) was added dropwise to a stirred suspension of 2-bromoethylamine hydrobromide (20.5 g.), in methylene chloride (100 ml.). The mixture was stirred for 3 hr., then filtered. The filtrate was concentrated to a small volume, and the residue was distributed between ether and water. The ether layer was dried, then distilled under reduced pressure, collecting the fraction b.p. 92.degree.-94.degree./0.9 nm as N-carbo-t-butoxy-2-bromoethylamine (1.756 g.)
PREPARATION 30
Pyrid-2-ylmethoxyimino(thien-2-yl)acetic acid (synisomer)
2-Chloromethylpyridine (a 25% solution in toluene, 2.8 ml.) was added to a solution of the sodium salt of methyl syn-hydroxyimino-thien-2-ylacetate (1.035 g.) in benzene:dimethylformamide (30 ml, 2:1, v/v). The solution was stirred for 18 hr., ethyl acetate (50 ml.) was added, and the mixture was washed several times with water, dried and evaporated to dryness to give a dark-green oil (1.4 g.). This oil was chromatographed on two 40.times. 20 cm. preparative chromatography plates, eluting with chloroform. The single major band was eluted off the silica with chloroform: ethanol (9:1 v/v) to give after evaporation of the solvent, methyl pyrid-2-ylmethoxyimino(thien-2-yl)acetate (889 mg., 47%) (75% syn, 25% anti-isomer). A solution of the crude ester (828 mg.) in methanol (20 ml) and 2 N sodium hydroxide solution (3 ml.) was stood 16 hrs. After removal of the methanol, the aqueous mixture was acidified to pH 2.0 in the presence of methylene chloride. The acid mixture was extracted with methylene chloride and the extracts were washed (water, brine), dried, and evaporated to dryness. Trituration of the residue with ether gave the title compound (210 mg, 27%), m.p. 152.1.degree.-152.9.degree., .lambda..sub.max, (EtOH) 260.5, 266,289 nm (.epsilon. 12,300; 12,000; 11,700), .tau. (DMSO-d.sub.6) values include 4.66 (singlet, CH.sub.2).
PREPARATION 31
n-Butoxyimino-thien-2-ylacetic acid (syn-isomer)
1-Bromobutane (0.6 ml.) was added to a solution of methyl syn-hydroxyimino-thien-2-ylacetate sodium salt (prepared by treating methyl syn-hydroxyimino-thien-2-ylacetate with 1 equivalent sodium methoxide) (1.0 g.) in benzene:dimethylformamide (2:1; 15 ml.) and the mixture was stirred for 17 hours at room temperature then poured into water. The aqueous solution was extracted with ethyl acetate, washed with water, dried and evaporated to give syn methyl ester (0.88 g) as a pale yellow oil.
2 N-Sodium hydroxide (4.0 ml.) was added to a solution of the syn methyl ester (0.85 g.), in methanol (10 ml.) and the mixture was left at room temperature for 18 hr. The methanol was removed by evaporation; the aqueous residue was diluted with water, washed with ether and acidified to pH 2.0 with 2 N-hydrochloric acid. The mixture was extracted with ethyl acetate; the combined extracts were washed with water, dried and evaporated to give the title compound (0.74 g., 81%) as a pale yellow oil, .tau. values (DMSO-d.sub.6) include 2.30 2.7-3.0 (thien-2-yl protons), 5.84 (OCH.sub.2), 9.10 (CH.sub.3).
PREPARATION 32
2-Methoxymethoxyimino-(thien-2-yl)acetic acid (syn-isomer)
A solution of sodium methoxide in methanol (approx. 0.2 M) was added to methyl 2-hydroxyimino-2-(thien-2-yl) acetate (syn-isomer) (0.5 g) and the solution formed was evaporated to a yellow oil which on azeotroping with petrol (b.p. 40.degree.-60.degree.) gave the sodium salt of methyl 2-hydroxyimino-2-(thien-2-yl) acetate (syn-isomer) (0.49 g., 88%). To a stirred solution of the sodium salt (0.49 g.) in benzene/DMF (5 mls., 2:1). was added chlorodimethyl ether (0.22 mls). After 10 mins. the reaction was poured into saturated sodium bicarbonate solution and extracted with benzene. The combined extracts were washed with water, dried over sodium sulphate and evaporated to a yellow oil, methyl 2-methoxymethoxyimino-(thien-2-yl) acetate (syn-isomer) (0.62 g; 100%), .lambda..sub.max. (EtOH) 288 nm (.epsilon. 10,300), .nu..sub.max. (CHBr.sub.3) 1730 (COOCH.sub.3), 1660 cm..sup.-.sup.1 (--C=N--), .tau. (CDCl.sub.3) values include 2.61-2.83 (multiplet, thien-2-yl), 4.83 (singlet, CH.sub.2), 6.06 (CO.sub.2 CH.sub.3) 6.56 (singlet, CH.sub.2 OCH.sub.3).
A solution of sodium hydroxide (4 ml, 2 N) and methanol were added to the methyl ester (0.4 g.). After 30 mins. the reaction was poured into water and washed with ethyl acetate. The aqueous layer was acidified to pH 1 using 2 N-hydrochloric acid and extracted with ethyl acetate. The ethyl acetate was dried and evaporated to a colourless oil which was azeotroped with petrol (bp. 40.degree.-60.degree.) to yield a white solid, syn 2-methoxymethoxyimino-(thien-2-yl)acetic acid (0.21 g; 55%), m.p. 61.2.degree., .lambda..sub.max. (EtOH) 286 nm (.epsilon. 10,400), .nu..sub.max. (Nujol) 1732, 2600 cm..sup.-.sup.1 (CO.sub.2 H), .tau. (DMSO-d.sub.6) 2.26 (multiplet; thienyl H.sub.5), 2.7-2.9 (multiplet, thienyl H.sub.3 and H.sub.4), 4.88 (singlet, O--CH.sub.2 --), 6.6 (singlet, OCH.sub.3).
PREPARATION 33
2-t-Butoxyiminobenzo[b]-thien-2'-ylacetic acid (syn-isomer)
Benzo[b]-thien-2-ylglyoxylic acid (3.09 g) and t-butoxyaminohydrochloride (1.98 g) were dissolved in 50% aqueous ethanol (100 ml). The solution was adjusted to pH 4.5 with sodium hydroxide solution and maintained at such for 4 hr. at room temperature. Thin-layer chromatography showed incomplete reaction. t-Butoxyaminohydrochloride (500 mg.) was added and the solution kept at room temperature overnight. The alcohol was removed by evaporation and the aqueous phase adjusted to pH 8 and washed with ether. The aqueous phase was then acidified to pH 1.5 under ether. The ether solution was washed with water and dried. Evaporation gave a cream solid (4.05 g.). Fractional crystallisation from cyclohexane gave the anti-isomer of the title compound (1.6 g.). The mother liquors were combined and evaporated to give a cream solid (2.11 g.) that was treated in ether with excess diazomethane in ether at 0.degree.-5.degree.. The excess reagent was destroyed by acetic acid and the ether solution washed with sodium bicarbonate, water and dried. Evaporation gave an oil (1.75 g.). This was dissolved in methanol (70 ml) and treated with sodium hydroxide solution (N: 7 ml.) at room temperature for 3 hr. Hydrochloric acid (2 N: 3.5 ml) was added and the methanol was removed by evaporation. The aqueous residue was partitioned between ether and sodium bicarbonate solution. The ether layer was washed with water and dried. Evaporation gave an oil (0.92 g.) that was dissolved in methanol (20 ml.) and treated with sodium hydroxide (N: 7 ml) at reflux temperature for 3 hr. Sodium hydroxide (N: 5 ml) was added and the solution refluxed for 6 hr. The methanol was removed by evaporation and the residue partitioned between ether and water. The aqueous phase was acidified (pH 1.5) under ether and the ether layer washed with water, dried and evaporated to give a pale orange crystalline solid (760 mg. 18%). Crystallisation from benzene containing cyclohexane gave syn-t-butoxyiminobenzo[b] -thien-2-ylacetic acid (430 mg.) m.p. 108.degree.-9.degree., .lambda..sub.max. (EtOH) 231, 253, 297 nm (.epsilon. 17,000, 7,240, 24,500).
PREPARATION 34
Benzo[b]-thien-2-ylglyoxylic acid and Benzo[b]-thien-3-ylglyoxylic acid
A mixture of 2- and 3-acetylbenzo[b]-thiophene (ca. 1:1) (11.0 g.) in pyridine (80 ml.) was warmed to 60.degree. with vigorous stirring and selenium dioxide (9.92 g.) was added portionwise. The mixture was heated to 110.degree. C. and an exothermic reaction occurred, the temperature rising to 120.degree.. The reaction was stirred at 90.degree. for 45 mins. and then left to cool. Water (80 ml.) was added and the mixture filtered through a kieselguhr pad. The pyridine was removed by evaporation and the aqueous residue again filtered. The filtrate was acidified to pH 2 under ether with 40% orthophosphoric acid (40 ml).
The aqueous phase was extracted with ether and the ether fractions were combined, washed with water and dried. Evaporation gave an orange crystalline solid (11.0 g., 86%). Crystallisation from benzene (100 ml) gave bright yellow crystals of benzo[b]-thien-2ylglyoxylic acid (2.3 g. 18%), m.p. 175.9.degree., .lambda..sub.max. (EtOH), 233, 247, .lambda. infl. 308 nm. (.epsilon. 11,400; 7,200; 14,600), .tau. (DMSO-d.sub.6) values include 1.83 (C-4 and C-7 protons), 1.42 (C-3 proton), 2.40 (C-5 and C-6 protons).
The mother liquor was concentrated to an orange oil which crystallised on standing (8 g.). Recrystallisation from benzene (20 ml.) gave pale yellow needles of benzo-[b]-thien-3-ylglyoxylic acid (1.6 g., 12.5%), m.p. 92.degree.-93.degree., .tau. (DMSO-d.sub.6) values include 0.83 (C-2 proton), 1.32 (C-4 proton), 1.79 (C-7 proton), 2.40 (C-5 and C-6 protons), .lambda..sub.max. (EtOH) 235; 310.5 nm (.epsilon. 11,200 and 7,400).
2-ALKOXYIMINOARYLACETYL CHLORIDES
PREPARATION 35
syn-2-Methoxyiminophenylacetyl chloride
Phosphorous pentachloride (5.21 g) was added in portions to a stirred suspension of syn-2-methoxyiminophenylacetic acid (4.51 g.) in dry benzene (20 ml.). Thionyl chloride (0.3 ml) was added to the solution, which was refluxed for 30 minutes. Benzene was removed by evaporation, and the residue distilled, producing a mixture of syn- and anti- acid chlorides (ca. 1:1) as a colourless oil (3.08 g., 62%), b.p. 74.degree. (0.01 nm). A repeat of this reaction (on 5.04 mmole) at room temperature also produced a mixture of the isomeric acid chlorides.
The acid chlorides were separated and purified by preparative plate chromatography, developing three times with petroleum spirit (b.p. 60.degree.-80.degree.) producing the title compound as a colourless oil (1.43 g. 24%).
In a further experiment a mixture of syn- and anti-2-methoxyiminophenylacetic acids (10 g., ca 1:1) were converted to a mixture of acid chlorides as above and chromatographed on silica gel (120 g., Hopkins and Williams, MFC) using petroleum spirit (b.p. 60.degree.-80.degree.) to give syn-2-methoxyiminophenylacetyl chloride (4.32 g., 39%).
General Method for Converting a 2-Alkoxyiminoarylacetic Acid into its Acid Chloride without Isomerisation
A solution of the pure syn- or 2-alkoxyiminoarylacetic acid (1 equiv.) in methanol (ca. 2-4 ml./mmole.) was treated with sodium methoxide (1 equiv.) in methanol at 0.degree.-25.degree. and the mixture evaporated to give the sodium salt which may be dried by azeotroping with several portions of benzene and/or drying in vacuo over phosphorus pentoxide.
The anhydrous sodium salt (1 equiv.) is suspended in dry benzene (ca. 5 ml/mmole) containing a few drops of dry dimethylformamide and treated with freshly distilled oxalyl chloride (1-2.5 equiv.). The mixture is stirred at room temperature for 1 hr. and then evaporated to remove benzene. The resulting acid chlorides were not characterised but were dissolved in acetone or methylene chloride and used immediately to acylate the appropriate cephalosporin nucleus.
The following acids were converted into their acid chlorides in this way:
Syn-2-Ethoxyiminophenylacetic acid,
Syn-2-t-Butoxyiminophenylacetic acid,
Syn-2-Benzyloxyiminophenylacetic acid,
Syn-2-Then-2'-yloxyiminophenylacetic acid,
Syn-2-Methoxyimino-(thien-2-yl)acetic acid,
Syn-2-Ethoxyimino-(thien-2-yl)acetic acid,
Syn-2-Benzyloxyimino-(fur-2-yl)acetic acid,
Syn-2-n-Butoxyimino-(thien-2-yl)-acetic acid,
Syn-2-t-Butoxyimino-(thien-2-yl)-acetic acid,
Syn-2-(2-Bromoethoxy)imino-(thien-2-yl)-acetic acid,
Syn-2-(2-t-Butoxycarbonylaminoethoxy)imino-thien-2-yl)acetic acid,
Syn-2-Benzyloxyimino-(thien-2-yl)acetic acid,
Syn-2-Then-2'-yloxyimino-(thien-2-yl)-acetic acid,
Syn-2-(1-Ethoxy)ethoxyimino-(thien-2-yl)-acetic acid,
Syn-2-(Pyrid-2-ylmethyl)-oxyimino-(thien-2-yl)acetic acid,
Syn-2-Methoxyiminonaphth-1'-ylacetic acid,
Syn-2-t-Butoxyiminonaphth-1'-ylacetic acid,
Syn-2-Benzyloxyiminonaphth-1'-ylacetic acid,
Syn-2-Methoxyiminobenzo-[b]-thien-3'-ylacetic acid,
Syn-2-t-Butoxyiminobenzo-[b]-thien-3'-ylacetic acid,
Syn-2-Benzyloxyiminobenzo-[b]-thien-3'-ylacetic acid,
Syn-2-Methoxyiminobenzo-[b]-thien-2'-ylacetic acid,
Syn-2-t-Butoxyiminobenzo-[b]-thien-2'-ylacetic acid,
Syn-2-Benzyloxyiminobenzo-[b]-thien-2'-ylacetic acid,
Syn-2-Methoxyimino(fur-2-yl)acetic acid,
Syn-2-t-Butoxyimino-(fur-2-yl)acetic acid,
Syn-2-Benzyloxyimino-(fur-2-yl)-acetic acid,
Syn-2-Furfuryloxyimino-(fur-2-yl)-acetic acid, and
Syn-2-ethoxyimino-(fur-2-yl)-acetic acid.
PREPARATION A
Then-2-yloxamine hydrochloride (used as a starting material in Preparation 7)
(a) N-(Then-2-yloxy)phthalimide
Anhydrous potassium carbonate (11.04 g.) was added to a stirred suspension of N-hydroxyphthalimide (17.12 g.) in dry dimethyl sulphoxide (200 ml.). A brown colour developed, 2-Chloromethylthiophene (28.5 g.) was added dropwise and the mixture was stirred for 16 hr., during which time the colour disappeared. The suspension was poured into water (800 ml.) and cooled to 5.degree.. The white precipitate was filtered off, and recrystallised from ethanol to give colourless needles of N-(then-2-yloxy)phthalimide (23.4 g., 83%), m.p. 129.7.degree.-130.9.degree. .tau. values (DMSO-d.sub.6) are 4.58 (CH.sub.2), 2.28, 2.68, 2.90 (thienyl protons) 2.08 (phthalimide protons)
(b) Then-2-yloxamine hydrochloride
A mixture of N-(then-2-yloxy)phthalimide (22.4 g.) 100% hydrazine hydrate (5 g) and ethanol (600 ml.) was heated under reflux for two hours. Initially, a yellow solution was formed, but soon solid began to precipitate. The mixture was cooled, then acidified with concentrated hydrochloric acid (12 ml.). The precipitated phthalhydrazide was filtered off and washed with ethanol (3.times. 50 ml.) and water (100 ml.). The combined filtrate and washings were evaporated to dryness, and the residue, suspended in water, was basified with 2 N sodium hydroxide solution. The basic mixture was extracted with ether, and the combined extracts were washed (water, saturated brine), dried, and saturated with dry hydrogen chloride. The precipitated solid was collected and well washed with ether to give then-2-yloxamine hydrochloride, (12.45 g., 87%), m.p. 157.1- 157.5.degree.. A sample recrystallised from ethanol/ether had m.p. 161.7- 162.1 .tau. values (DMSO-d.sub.6) include 4.69 (CH.sub.2), 2.30, 2.72, 2.90 (thienyl protons)
PREPARATION B
Furfuryloxamine Hydrochloride (used as a starting material in Preparation 22)
(a) N-Furfuryloxyphthalimide
To a stirred mixture of N-hydroxyphthalimide (41 g.), anhydrous potassium carbonate (26.4 g.) and dry dimethyl sulphoxide (400 ml.) was added 2-chloromethylfuran (freshly prepared, but undistilled, from 46.2 g. furfuryl alcohol according to the method of W. R. Kirner JACS, 1928, 50, 1955). The mixture was stirred for 18 hr., then poured into water (1.5 l). The precipitated solid was filtered off, washed well with water, and recrystallised from ethanol to give N-furfuryloxyphthalimide (42.8 g., 70%), m.p. 145.3.degree.-146.2.degree. .tau. values (DMSO-d.sub.6) are 4.80 (CH.sub.2), 2.22, 3.30, 3.50 (furyl protons) 2.08 (phthalimide protons).
(b) Furfuryloxamine Hydrochloride
100% Hydrazine hydrate (20 ml.) was added to a stirred solution of N-furfuryloxyphthalimide (42.0 g.) in methylene chloride (600 ml.). A copious precipitate formed immediately, and the mixture was stirred for 45 min. 5 N Ammonium hydroxide solution (500 ml.) was added to dissolve the precipitate, the two layers were separated, and the aqueous layer was washed twice with methylene chloride. The combined methylene chloride extracts were washed (saturated brine) and dried. Methylene chloride was evaporated off, and the residual liquid was dissolved in ether (250 ml.). Dry hydrogen chloride was passed into this solution for one hour. The precipitated solid was filtered off, washed with ether, dried, and recrystallised from isopropanol to give furfuryloxamine hydrochloride (12.89 g., 50%), m.p. 135.degree.-136.degree. (decom) .tau. values (DMSO-d.sub.6) include 4.87 (CH.sub.2), 2.20, 3.27, 3.44 (furyl protons).
PREPARATION C
The general procedures described in Preparations 5-23 for the preparation of 2-alkoxyiminoarylacetic acids were employed to prepare the intermediates listed in tabular form below (the Table may be regarded as a continuation of Table 1).
2-Alkoxyiminoarylacetyl Chlorides (continued)
The following acids were converted into their acid chlorides using the general method for converting a 2-aklkoxyimino arylacetic acid into its acid chloride without isomerisation described immediately after preparation 35:
Syn-2-Butoxyiminophenylacetic acid
Syn-2-Isopropoxyiminophenylacetic acid
Syn-2-Propoxyiminophenylacetic acid
Syn-2-Propoxyimino-(thien-2-yl)acetic acid
Syn-2-Ethoxyimino (benzo[b]-fur-2-yl)acetic acid
Syn-2-Ethoxyimino-(thien-3-yl)acetic acid
PREPARATION 40
2-Phenoxyimino-2-phenylacetic acid (syn- isomer)
A solution of syn-2-hydroxyimino-2-phenylacetic acid (33 g) in dry methanol (500 ml) was treated with 1.105 N sodium methoxide solution (486 ml), and stirred for 15 minutes. To the solution was added diphenyliodonium bromide (90 g), and the resulting mixture was stirred for 18 hours under nitrogen. A small amount of solid was filtered off, and the filtrate was evaporated to dryness. Water (600 ml) and ether (600 ml) were added to the residue, and the pH of the mixture was adjusted to 7.0 with concentrated hydrochloric acid. The aqueous layer was washed twice with ether, and then acidified under ether to pH 1.8 with concentrated hydrochloric acid. The acid mixture was extracted into ether, and the combined extracts were washed (water, saturated brine), dried, and evaporated to give a dark brown solid (ca 35 g). This solid was triturated with ice-cold nitromethane. The solid was collected, washed with a little cold nitromethane, and dried in vacuo to give fawn crystals of the title acid (24.41 g, 51%), m.p. 104.8.degree.-105.1.degree., .lambda..sub.max. (ethanol) 267.5, 285 nm (.epsilon. 11,600; 10,100).
Similarly were prepared:
PREPARATION 41
2-Phenoxyimino-2-(thien-2-yl)acetic acid (syn-isomer) (52%) m.p. 98.3.degree.-99.5.degree., .lambda..sub.max. (ethanol) 267.5, 303 nm. (.epsilon. 9,900; 12,000).
and
PREPARATION 42
2-Phenoxyimino-2-(fur-2-yl)acetic acid (syn-isomer) (34%), m.p. 100.7.degree.-100.9.degree., .lambda..sub.max. (ethanol) 270.5, 292.5 nm (.epsilon. 14,300; 15,700).
PREPARATION 43
2-Cyclopentyloxyimino-2-(fur-2-yl)acetic acid (syn-isomer)
Fur-2-ylglyoxylic acid (2.80 g) and cyclopentyloxamine hydrochloride (3.3 g) were dissolved in a mixture of water (100 ml) and ethanol (50 ml), and the pH of the solution was adjusted to 5.0. The solution was stirred for 19 hours, the alcohol was evaporated off, and the solution was acidified to pH 1.5 under ethyl acetate. The acid mixture was extracted into ethyl acetate, and the combined extracts were washed, dried, and evaporated to give the crude acid (4.38 g). This acid was treated with charcoal in benzene for 15 minutes, filtered, and the filtrate was evaporated to give a solid, which was recrystallised twice from cyclohexane to give the title acid (2.28 g, 51%), m.p. 96.6.degree.-97.7.degree., .lambda..sub.max. (ethanol) 277.5 nm (.epsilon. 15,600).
PREPARATIONS 44-50
2-Alkoxyimino-2-arylacetic acids
General Procedures
A mixture of the substituted glyoxylic acid and an excess (10 to 15%) of the alkoxyamine hydrochloride was suspended in water or aqueous ethanol, stirred, and the pH of the mixture adjusted to between 4 and 5 with sodium hydroxide solution (N to 10 N). A clear solution at pH 4 to 5 was maintained during the reaction by further additions of sodium hydroxide solution and ethanol as needed. The reaction mixture was kept at room temperature until all of the keto-acid was consumed (it may be necessary to add a further portion of the more volatile alkoxyamines). The progress of the reaction was followed by acidification of an aliquot, extraction with ethyl acetate and thin layer chromatography of the extract on silica plates (developed with a mixture of chloroform:methanol:acetic acid; 18:2:1). The alkoxyiminoacetic acids were less polar than the starting keto-acids. The reaction times were 2 hours to 2 days. When reaction was complete the pH of the mixture was adjusted to between 7 and 8 and the ethanol (if any) was removed by evaporation. The aqueous mixture was extracted with ether, the extract discarded and the aqueous phase acidified to pH <2 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate or ether, the extract dried and evaporated to give the crude product which was purified by one of the following methods:
(a) Crystallisation and recrystallisation (if needed) from a suitable solvent.
(b) The crude product dissolved in ether was treated with a small excess of a solution of diazomethane in ether. The excess reagent was destroyed with acetic acid and the solution washed with sodium bicarbonate solution and evaporated to give the crude methyl esters. The esters were separated by preparative thick layer chromatography or column chromatography on silica, and then hydrolysed conventionally with alkali to give the syn acids, which were purified by crystallisation from a suitable solvent.
These methods were employed to prepare the intermediates listed in Table 4 (syn-isomers).
PREPARATION 51
2-(Thien-2-ylmethoxyimino)-1-methylpyrrol-2-yl)acetic acid (syn-isomer)
A solution of 1-methylpyrrol-2-ylglyoxylic acid (4.6 g) and thien-2-ylmethoxamine hydrochloride (5.46 g) in aqueous ethanol (100 ml, 1:1) was adjusted to pH 4.8 with 10 N-sodium hydroxide solution and stirred at pH 4.8 for 24 hours at room temperature. A further portion of thien-2-ylmethoxamine (0.5 g) was added and the solution was maintained at pH 4.8 and room temperature for a further 2 days. The pH was then adjusted to 8 with sodium bicarbonate solution and the ethanol was removed by evaporation. The aqueous residue was washed with ether and the aqueous phase was acidified to pH 1.5 under ether with 2 N-hydrochloric acid. The ether extracts were combined and washed with water, dried and evaporated to give an orange oil (8.8 g). The crude mixture of syn and anti-isomer was esterified with a slight excess of diazomethane in ether.
To a solution of the mixed methyl esters (7.7 g) in methanol (100 ml) was added N-sodium hydroxide (28 ml). The mixture was kept at room temperature for 3 hours when thin-layer chromatography of an aliquot showed only traces of remaining anti-ester. After a further 30 minutes 2 N-hydrochloric acid (14 ml) was added and the methanol was removed by evaporation. The residue was partitioned between ether and excess sodium bicarbonate in water. The ether layer was separated, washed with water, dried and evaporated to a pale orange oil (5.9 g).
This oil in methanol (100 ml) was treated with 10 N-sodium hydroxide solution (4.5 ml) and kept at room temperature for 16 hours. A further portion of 10 N-sodium hydroxide solution (4.5 ml) was added and after 24 hours at room temperature the mixture was warmed to 60.degree. for 30 minutes. The methanol was removed by evaporation and the residue divided between ether and sodium bicarbonate solution. The aqueous phase was acidified under ether with 2 N-hydrochloric acid. The combined ether extracts were washed with water and dried. Evaporation of the ether gave a pale orange oil (4.8 g) which was crystallised from carbon tetrachloride to give the title compound as pale brown crystals (1.9 g); m.p. 70.degree.-71.degree.; .lambda..sub.max. (EtOH) 235, 287.5 nm (.epsilon. 11,600 and 17,100): .tau. (DMSO-d.sub.6) values include 4.70 (S, CH.sub.2) and 6.18 (S, CH.sub.3).
GENERAL METHOD FOR CONVERTING A 2-SUBSTITUTED OXIMINO-2-ARYLACETIC ACID INTO ITS ACID CHLORIDE WITHOUT ISOMERISATION
A solution of the pure syn-2-substituted oxyimino-2-arylacetic acid (1 equiv.) in methanol (ca. 2-4 ml/mmole) was treated with sodium methoxide (1 equiv.) in methanol at 0.degree.-25.degree. and the mixture evaporated to give the sodium salt which may be dried by azeotroping with several portions of benzene and/or drying in vacuo over phosphorus pentoxide.
The anhydrous sodium salt (1 equiv.) was suspended in dry benzene (ca. 5 ml/mmole) containing a few drops of dry dimethylformamide and treated with freshly distilled oxalyl chloride (1-2.5 equiv.). The mixture was stirred at room temperature for 0.5-1 hour and then evaporated to remove benzene. The resulting acid chlorides were not characterised but were dissolved in acetone and used immediately to acylate the appropriate cephalosporin nucleus.
The acids described in Preparations 40-51 were converted into their acid chlorides in this way.
PREPARATION 52
Cyclopentyloxamine hydrochloride
A mixture of bromocyclopentane (14.9 g), N-hydroxyphthalimide (16.3 g), triethylamine (15 ml), and dimethylformamide (30 ml) was stirred for 16 hours, then poured into water (500 ml). The oily mixture was extracted with ethyl acetate, and the combined extracts, after washing (water), drying, and removal of solvent gave a white solid. This solid was recrystallised from ethanol to give N-cyclopentyloxyphthalimide (11.37 g, 49%); m.p. 81.2.degree.-82.5.degree.; .nu..sub.max. (CHBr.sub.3) include 1780, 1720 cm.sup..sup.-1 (CO--N--CO), 970 cm.sup..sup.-1 (>N--O--<CH); .tau. values (DMSO-d6) 2.08 (4 Ar-H), 5.12 (cyclopentyl 1-H), 8.18 (4-CH.sub.2).
A mixture of N-cyclopentyloxyphthalimide (11 g), 100% hydrazine hydrate (2.6 g), and ethanol (30 ml) was heated under reflux for 5 minutes. Concentrated hydrochloric acid (6 ml) was added to the mixture, which was heated under reflux for a further 5 minutes. Water (20 ml) was added to the mixture, which was cooled to room temperature, and filtered. The filtrate was evaporated to dryness, ethanol (50 ml) was added to the residue, and a small amount of insoluble material was filtered off. The filtrate was evaporated to dryness, and the residue was recrystallised from ethanol/ether to give cyclopentyloxamine hydrochloride (6.28 g, 96%), m.p. 156.9.degree..
PREPARATION 53
t-Butoxycarbonylmethoxamine
t-Butyl chloroacetate (13.0 g, prepared according to Org. Synth., Coll. Vol. 4, 263) was added dropwise to a stirred mixture of N-hydroxyphthalimide (14.2 g), triethylamine (23.0 g), and dimethylformamide (30 ml), and the resulting mixture was stirred for 4 hr. The mixture was poured into water (500 ml), and the precipitated solid was collected, washed with water, and dried. Recrystallisation from ethanol gave N-t-butoxycarbonylmethoxyphthalimide (17.26 g, 72%); m.p. 145.6.degree.; .tau. values (DMSO-d6) 2.09 (4 Ar-H), 5.28 (CH.sub.2), 8.56 (Bu.sup.t).
A solution of N-t-butoxycarbonylmethoxyphthalimide (21 g) in methylene chloride (250 ml) was treated with 100% hydrazine hydrate (7.6 ml) in methanol (15 ml), and the mixture was stirred for 1.5 hr. 5 N-Ammonia solution was added to dissolve the precipitated solid. The organic layer was separated, and the aqueous layer was further extracted with methylene chloride. The combined extracts were washed with water, dried, and evaporated to give a pale-yellow solid. To this was added ether, the mixture was filtered, and the filtrate evaporated to give t-butoxycarbonylmethoxamine as a pale yellow liquid, (8.88 g, 80%); .nu..sub.max. (Nujol) includes 3330, 3260 cm..sup..sup.-1 (NH.sub.2), 1742 cm.sup..sup.-1 (--COOBu.sup.t); .tau. values (DMSO-d6) are 3.75 (--NH.sub.2), 5.96 (CH.sub.2) 8.55 (Bu.sup.t).
PREPARATION 54
Methyl N-Benzyloxymethylpyrrol-2-ylglyoxylate
Methyl pyrrol-2-ylglyoxylate (306 mg) in diglyme was treated with sodium hydride (63 mg) and stirred at room temperature for 3 hr. Benzyloxymethyl chloride (376 mg) was added and the mixture stirred at room temperature for a further 3 hr. The suspension was filtered and the filtrate evaporated. The residue, in ether, was washed with sodium bicarbonate solution, water and dried. Evaporation gave the crude product as a brown oil (530 mg). Purification by preparative thin-layer chromatography gave the title compound as a colourless oil (250 mg, 46%); .tau. values (DMSO-d6) include 6.10 (--CH.sub.3), 4.20 (>N--CH.sub.2 --), 5.47 (--OCH.sub.2 --), 2.70 (--Ph).
PREPARATION 55
N-Benzyloxymethylpyrrol-2-ylglyoxylic acid
Crude methyl-N-benzyloxymethylpyrrol-2-ylglyoxylate (10 g) in methanol (150 ml) was treated with sodium hydroxide solution (N: 40 ml) at room temperature for 1 hr. Thin-layer chromatography showed complete hydrolysis. Hydrochloric acid (2N: 20 ml) was added and the methanol was removed by evaporation. The residue was shaken with sodium bicarbonate solution and ether. The aqueous layer was acidified under ether and the ether extract was washed with water and dried. Evaporation gave the acid as an orange oil (4.5 g, 55%); this material was used directly to make the syn-methoxime described in Preparation 49.
PREPARATION 56
syn-Isopropoxyiminophenylacetic acid
A mixture of phenylglyoxylic acid (3.0 g.), isopropoxyamine hydrochloride (2.5 g.), ethanol (100 ml) and water (50 ml) was stirred and adjusted to pH 4.5 to 5 with sodium hydroxide solution (2 N). The solution was stirred for 5 hr. maintaining the pH at 4.5-5 with further additions of sodium hydroxide solution. The ethanol was removed by evaporation, the aqueous residue acidified and the product collected by extraction with ethyl acetate. Evaporation of the ethyl acetate gave a brown oil (4.2 g.) that was esterified conventionally with diazomethane to give a mixture of the syn and anti methyl esters of the title compound as an oil (4.04 g.).
The mixture of ester (4.0 g) in methanol (60 ml) was treated with sodium hydroxide solution (2 N:19.0 ml) and kept for 2 hr. at room temperature. The methanol was evaporated and the residue, diluted with water, extracted with ethyl acetate. Evaporation of the dried (MgSO.sub.4) ethyl acetate solution gave the crude syn methyl ester (0.82 g.). The ester (0.82 g) in methanol (20 ml) was treated with sodium hydroxide solution (2 N:3.6 ml) and kept at room temperature for 31 hr. Conventional isolation of acidic material gave the crude syn isomer (0.706 g) which was recrystallised from cyclohexane to give the title compound (0.358 g.) m.p. 59.5.degree. .lambda..sub.max. (EtOH) 258 nm (.epsilon.12,700), .tau.(DMSO-d.sub.6) values include 2.47 (phenyl), 5.53 (O--CH< ), 8.71 (CH.sub.3).
PREPARATION 57
syn-Propoxyiminothien-2-ylacetic acid
A mixture of thien-2-ylglyoxylic acid (3.12 g), n-propoxyamine hydrochloride (2.8 g.), ethanol (75 ml) and water (75 ml) was adjusted to pH 4.5 to 5 with sodium hydroxide solution (2 N) and stirred at room temperature. A clear solution at pH 4.5 to 5 was maintained by further additions of base and ethanol as required. After 4 hr. a further portion of n-propoxyamine hydrochloride (1.4 g) was added and the mixture stirred for a further 3 hr. (keeping the pH at 4.5-5) and then kept overnight. The ethanol was evaporated and the residual solution diluted with water, acidified and extracted with ethyl acetate, Evaporation of the dried (MgSO.sub.4) ethylacetate solution gave a mixture of the syn and anti forms of the title acid as an oil (4.8 g.).
The mixture of acids was esterified conventionally with diazomethane to give a mixture of the syn and anti methyl ester (3.175 g.).
The mixture of esters in methanol (50 ml.) was treated with sodium hydroxide solution (2 N; 14 ml) for 10 min. at room temperature. The methanol was removed, rapidly, by evaporation and the residue, in water, extracted with ethyl acetate. Evaporation of the dried (MgSO.sub.4) ethyl acetate solution gave the syn methyl ester (0.416 g.). The ester in methanol (10 ml) was treated with sodium hydroxide solution (2 N: 1.7 ml) and kept at room temperature for 26 hr. Conventional isolation of acid material gave the title compound as an oil (0.235 g.) .tau. (DMSO-d.sub.6) values include 2.28, 2.7-2.9 (thienyl), 5.90 (O--CH.sub.2).
EXAMPLE 1
3-Acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)-ceph-3-em-4-carb oxylic acid (syn-isomer)
To a solution of dicyclohexylcarbodiimide (1.15 g.) and t-butyl 3-acetoxymethyl-7.beta.-aminoceph-3-em-4-carboxylate (1.83 g.) in dry methylene chloride (30 ml) was added a solution of syn 2-methoxyiminophenylacetic acid (1 g.) in dry methylene chloride (15 ml.) and the resulting solution was stirred at room temperature for 1.5 hours. After filtration the filtrate was washed with 2 N hydrochloric acid, water, saturated sodium bicarbonate solution, water dried, and evaporated to an orange froth (2.60 g.). Part of this froth (0.21 g.) was triturated with petroleum spirit (b.p. 60.degree.-80.degree.) producing syn-t-butyl 3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido) ceph-3-em-4-carboxylate as a solid (0.15 g.), .lambda..sub.max. (EtOH) 258 nm. (.epsilon. 17,400), .nu..sub.max. (CHBr.sub.3) 1784 cm..sup..sup.-1 (.beta.-lactam), .tau. (CDCl.sub.3) 2.99 (doublet J 9.0; NH), 2.2-2.7 (multiplet; Ph), 5.97 (singlet; OCH.sub.3) 7.93 (singlet; acetate), 8.48 (singlet; t-butyl).
The remainder of the froth (2.4 g.) was shaken with trifluoroacetic acid (10 ml) at room temperature for 10 minutes, and evaporated to an oil. Ether (.about.80 ml) was added, producing a solution, which on standing precipitated syn-3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4- carboxylic acid as a white solid (1.58 g., 66%), [.alpha.].sub.D.sup.23 +55.degree. (c, 1.02 in dioxan), .lambda..sub.max. (pH 6 phosphate buffer) 258 nm. (.epsilon.19,550), .lambda..sub.max. (Nujol) 1782 cm..sup..sup.-1 (.beta.-lactam), .tau.(DMSO-d.sub.6) 0.21 (doublet, J 8.0 Hz; NH), 2.3-2.6 (multiplet; Ph), 6.08 (singlet; OCH.sub.3), 7.98 (singlet, acetate).
EXAMPLE 2
3-Crotonoyloxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4- carboxylic acid (syn-isomer).
A suspension of diphenylmethyl 7.beta.-amino-3-crotonyloxymethylceph-3-em-4-carboxylate toluene-p-sulphonate salt (1.0 g) in ethyl acetate (20 ml) was shaken with sodium bicarbonate solution. The ethyl acetate layer was washed with water and brine, dried and concentrated under reducer pressure. The residue was dissolved in methylene chloride (10 ml) and dicyclohexylcarbodiimide (0.324 g) was added. This solution was treated with 2-methoxyimino-2-phenylacetic acid (syn-isomer) (0.281 g). After 1 hour the mixture was filtered, and concentrated under reduced pressure. The residue was dissolved in ethyl acetate and the solution was washed with 2 N-hydrochloric acid, sodium bicarbonate solution, water and brine, dried and concentrated under reduced pressure. The resulting foam (1.0 g) was dissolved in anisole (2 ml). After 5 minutes at 20.degree. the solution was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and this solution was repeatedly extracted with sodium bicarbonate solution. The combined extracts were washed with ethyl acetate and then acidified with 2 N-hydrochloric acid and extracted with ethyl acetate. The extract was washed with water and brine, dried, concentrated under reduced pressure and added to stirred petroleum to precipitate the title methoxime (0.467 g; 67%) [.alpha.].sub.D +37.3.degree. (c 0.9 in DMSO) .lambda..sub.max. (pH 6 phosphate buffer) 258 nm (.epsilon.17,800), .lambda..sub.max. (Nujol) 1782 (.beta.-lactam), 1710 (CO.sub.2 H), 1680 and 1540 (CONH) and 1660 cm..sup..sup.-1 (unsaturated ester), .tau. (DMSO-d.sub.d 6, 100 MHz) 0.19 (doublet, J 8 Hz); 3.03 (multiplet; CH=CH-- CH.sub.3) 6.06 (singlet; NOCH.sub.3), 6.28 and 6.50 [2 doublets (branches of quartet), J 18 Hz; C-2 CH.sub.2 ], and 8.12 (double doublets, J 7 and 1.5 Hz; CH=CH-- CH.sub. 3).
Examples 3-11
General Procedures for the Preparation of 7.beta.-(2-alkoxyimino-2-substituted-acetamido)-3-(substituted)methylceph- 3-em-4-carboxylic acids using Dicyclohexylcarbodiimide
(i) Preparation of Intermediate 4-Carboxylic Acid Esters
Method A
To a solution of a t-butyl or diphenylmethyl ester of 7.beta.-amino-3-(substituted)-methylceph-3-em-4-carboxylic acid (1 equiv.) and dicyclohexylcarbodiimide (1-1.2 equiv.) in dry methylene chloride (cosolvents e.g. dimethylformamide or dioxan have been used) was added, at 0.degree.-20.degree., a solution of the syn 2-alkoxyimino-substituted acetic acid (1 equiv.) in dry methylene chloride. After stirring for 45 min-3 hr. at room temperature the mixture was filtered, the filtrate was washed successively with 2 N-hydrochloric acid, saturated sodium hydrogen carbonate solution, water and brine. The organic phase was dried and evaporated to give the required ester as an oil or foam.
Method B
As above using the dimethyl ether of diethylene glycol in place of methylene chloride.
Method C
As method A with purification of the ester by chromatography on silica.
Method D
As Method A but with prior regeneration of the 7.beta.-amino ester from its p-toluene-sulphonic acid salt (1 equiv.) by shaking with ethyl acetate and an excess of saturated sodium bicarbonate solution. After washing with water and brine the organic layer was evaporated to dryness and redissolved in methylene chloride.
(ii) Deprotection of the Intermediate Esters
Method E
The t-butyl or diphenylmethyl ester was dissolved in trifluoroacetic acid (5-10 ml./gm. ester) and left at room temperature for (5-10 min.) then evaporated under reduced pressure. The crude product was taken up in ether or ethyl acetate, extracted into aqueous sodium hydrogen carbonate solution and washed with ethyl acetate. The aqueous layer was acidified with 2 N-hydrochloric acid and extracted into ethyl acetate. The organic layer was washed, dried and evaporated to give the required syn-7.beta.-(2-alkoxyimino-2-substituted acetamido)-3-(substituted)-methylceph-3-em-4-carboxylic acid.
Method F
The ester was dissolved in anisole (1- 5 ml/grm. ester) and treated with trifluoroacetic acid (4-10 ml./grm. ester) at room temperature for 5-10 min. then worked up as described in Method E.
(iii) Preparation of Diphenylmethyl 7.beta.-Amino-3-cyclopropylcarbonyloxymethylceph-3-em-4-carboxylate p-toluene-sulphonic acid salt used as Starting Material in Example 11
(a) Diphenymethyl 3-cyclopropylcarbonyloxymethyl-7.beta.-(thien-2-yl)-acetamidoceph-3-em-4-c arboxylate
To a stirred solution of diphenylmethyl 3-hydroxymethyl-7.beta.-(thien-2-yl)acetamidoceph-3-em-4-carboxylate (8 g.) in dry tetrahydrofuran (20 ml.) containing anhydrous pyridine (6.7 ml.) was added a solution of cyclopropane carbonyl chloride (from the acid, 6.7 g. and thionyl chloride at room temperature for 90 mins.) in dry tetrahydrofuran (10 ml.) over a period of 10 mins. at room temperature. After stirring at room temperature for 2 hr. the mixture was evaporated to near dryness and diluted with ethyl acetate (100 ml.). The mixture was washed successively with sodium hydrogen carbonate solution, 2 N-hydrochloric acid and water. The organic layer was stirred with charcoal at room temperature for 1 hr., filtered and the filtrate evaporated to a foam. Trituration with petroleum spirit (bp. 60.degree.-80.degree.) gave the title ester (7.8 g., 87%), [.alpha.].sub.D +28.degree. (c 0.83 in dioxan), .lambda. inf. (EtOH) 235, 255 nm (.epsilon.12,800; 7,700), .lambda..sub.max. (CHBr.sub.3) 1790 (.beta.-lactam), 1728 cm..sup..sup.-1 (esters), .tau. (DMSO-d.sub.6) values include 0.8 (doublet NH) 3.02 (CHPh.sub. 2), 8.4 (multiplet, ##STR71## 9.0- 9.2 (CH.sub.2 groups in the cyclopropane ring).
EXAMPLE 3
(a) t-Butyl 3-methyl-7.beta.-(2-methoxyimino-2-phenylacetamido)-ceph-3-em-4-carboxylat e(syn-isomer) was obtained as a yellow solid m.p. 66.degree. (dec.), .lambda..sub.max. (EtOH) 255 nm (.epsilon.16,350), .lambda..sub.max. (CHBr.sub.3) 1785 (.beta.-lactam), 1720 (t-butyl ester), 1690, 1520 cm..sup..sup.-1 (CONH); .tau. (CDCl.sub.3) 3.02 (doublet J 8.0 Hz; NH) 2.3-2.7 (multiplet, Ph), 5.96 (singlet; OCH.sub.3), 7.89 (singlet, CH.sub.3) 8.48, (singlet; t-butyl) by Method A.
Deprotection by Method E gave:
(b) 3-Methyl-7.beta.-(2-methoxyimino-2-phenylacetamido) ceph-3-em-4-carboxylic Acid (syn-isomer), as a cream solid (67%), m.p. 145.degree. (dec.), [.alpha.].sub.D.sup.17 +100.degree. (c 1.32 in dioxan) .lambda..sub.max. (pH 6.0 phosphate buffer) 258-259 nm (.epsilon.18,650), .lambda..sub.max (Nujol) 1760 (.beta.-lactam), 1702 (CO.sub.2 H), 1660. 1536 (CONH); .tau. (DMSO-d.sub.6) 0.24 (doublet J 8.0 Hz; NH), 2.3-2.6 (multiplet; Ph). 6.05 (singlet, OCH.sub.3), 7.94 (singlet; CH.sub.3).
EXAMPLE 4
(a) t-Butyl (3-Acetoxymethyl-7.beta.-(2-benzyloxyimino-2-phenylacetamido)ceph-3-em-4-c arboxylate(syn-isomer) m.p. 123.degree.-125.degree., .lambda..sub.max (EtOH) 259 nm (.epsilon.20,500), .lambda..sub.max (CHBr.sub.3) 1796 (.beta.-lactam), 1732 (acetate and ester), 1694, and 1522 cm..sup..sup.-1 CONH); (CDCl.sub.3) 3.00 (doublet J 9.0 Hz; NH) 2.38, 2,60 (multiplet, Ph), 2.60 (benzyl aromatic protons), 4.74 (singlet; methylene group of benzyl) 7.94 (singlet, acetate), 8.48 (singlet, t-butyl) was obtained by Method A. Deprotection by Method E gave:
(b) 3-Acetoxymethyl-7.beta.-(2-benzyloxyimino-2-phenylacetamido)-ceph-3-em-4-c arboxylic acid(syn-isomer) as a white solid (58%), m.p. 167.degree. (dec.) [.alpha.].sub.D.sup.26 +50.5.degree. (c, 0.85 in dioxan), .lambda..sub.max. (pH 6.0 phosphate buffer) 259 nm (.epsilon.20,700)., .nu..sub.max. (Nujol) 1771 (.beta.-lactam), 1735, 1252 (acetate), 1650 and 1530 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) 0.14 (doublet J 8.0 Hz; NH), 2.3-2.7 (multiplet; aromatic protons), 4.79 (singlet, CH.sub.2 of benzyl group), 7.96 (singlet, acetate).
EXAMPLE 5
(a) Diphenylmethyl 7.beta.-(2-methoxyimino-2-phenylacetamido)-3-methylthiomethylceph-3-em-4-c arboxylate(syn-isomer) .lambda..sub.max. (EtOH) 259 nm (.epsilon.18,550), .nu..sub.max. (CHBr.sub.3) 1785 (.beta.-lactam), 1722 (benzhydryl ester), 1688 and 1518 cm..sup..sup.-1 (CONH)l .tau. (CDCl.sub.3) 3.01 (doublet 7 Hz; NH), 2.32, 2.64, 2.65 (aromatic protons) 5.97 (singlet; OCH.sub.3), 8.16 (singlet, SCH.sub.3) was prepared by Method A. Subsequent deprotection by Method E gave:
(b) 3-Methylthiomethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)-ceph-3-em-4- carboxylic acid(syn-isomer) as a precipitated solid., (51%), [.alpha.].sub.D.sup.26 +51.degree. (c, 0 98 in dioxan), .lambda..sub.max. (pH 6.0 phosphate buffer) 259 nm, (.epsilon.18,300), .nu..sub.max. (Nujol) 1781 (.beta.-lactam), 1770 (CO.sub.2 H), 1670 and 1531 cm..sup..sup.-1 (CONH); .tau. (DMSO-d.sub.6) 0.22 (doublet J 9 Hz; NH), 6.06 (singlet OCH.sub.3), 8.01 (singlet SCH.sub.3).
EXAMPLE 6
(a) Diphenylmethyl 3-Benzoyloxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-c arboxylate(syn-isomer) was prepared as a crude orange solid (80%) by Method A. Deprotection by Method F gave:
(b) 3-Benzoyloxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)-ceph-3-em-4 -carboxylic acid(syn-isomer) as a white powder (48%) [.alpha..sub.D +46.5.degree. (c 0.9 in DMSO) .lambda..sub.max. (pH 6 phosphate buffer) 232.5 (.beta.22,400) and 258.5 nm (.delta.21.400 ) .nu..sub.max. (Nujol) 1780 (.beta.-lactam), 1720 and 1265 (benzoate), 1705 (CO.sub.2 H), 1675 and 1532 cm.sup..sup.-1 (CONH) .tau. (DMSO-d.sub.6, 100 MHz) 0.16 (doublet, J 8 Hz; NH) 4.06 (double doublet J 5 and 8 Hz; C-7H), 4.70 (doublet; J5 Hz; C-6 H), 6.03 (singlet; NOCH.sub.3), and 6.14 and 6.44 [2 doublets (branched of quartet), J 13 Hz; C-2 CH.sub.2 ] .
EXAMPLE 7
(a) Diphenylmethyl 3-Isobutyryloxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em- 4-carboxylate(syn-isomer) was prepared by Method A and deprotected without purification by Method F to give:
(b) 3-Isobutyryloxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em- 4-carboxylic acid(syn-isomer) (73%) [.alpha.].sub.D +48.5.degree. (c 1.0 on DMSO) .lambda..sub.max. (pH 6 phosphate) 258 nm (.epsilon.18,700), .nu..sub.max. (Nujol) 1783 (.beta.-lactam), 1725 (carboxylic ester and acid), 1670 and 1530 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6 ; 100 MHz) 0.19 (doublet, J 8 Hz; NH) 2.2-2.6 (multiplet; aromatic protons), 4.09 (doublet, J 5 and 8 Hz; C-7H), 4.76 (doublet, J 5 Hz; C-6 H), 4.94 and 5.26 [2 doublets (branches of quartet), J 13 Hz; C-3 CH.sub.2 ], 6.04 (singlet; NOCH.sub.3), 6.28+ 6.50 [2 doublets (branches of quartet), J 18 Hz; C-2 CH.sub.2 ], 7.40 (septet, J 7 Hz; CH(CH.sub. 3).sub.2) and 8.80 (doublet, J 7 Hz, CH(CH.sub.3).sub.2).
EXAMPLE 8
(a) t-Butyl 3-Acetoxymethyl-7.beta.-[2-methoxyimino-2-(thien-2-yl)-acetamido]-ceph-3-e m-4-carboxylate(syn-isomer) was obtained as a solid (85%), .lambda..sub.max..sup.EtOH 262-263 nm (.epsilon.14,900), .lambda..sub.inf. 280 nm, (.epsilon.13,280) .nu..sub.max. (CHBr.sub.3) 3400 (NH); 1780 (.beta.-lactam); 1738, 1120 (CO.sub.2 R) and 1684, 1514 cm..sup..sup.-1 (CONH) .tau. (CDCl.sub.3) 2.5-2.7, 2.95 (thienyl protons); 5.97 (OMe); 7.93 (OAc), and 8.48 (t-butyl) by Method A. Deprotection by Method E gave the free acid:
(b) 3-Acetoxymethyl-7.beta. -[ 2-methoxyimino-2-(thienyl-2-yl)-acetamido]-ceph-3-em-4-carboxylic acid(syn-isomer) (56%) [.alpha.].sub.D.sup.23 +60.degree. (c, 0.8 in dioxan), .nu..sub.max. 262 nm (.epsilon.15,700), .lambda..sub.inf. 290 nm. (.lambda.10,700), .nu..sub.max. 3275 (NH); 1768 (.beta.-lactam); 1728 (OAc); 1700, 2600 (CO.sub.2 H); and 1648, 1520 cm..sup..sup.-1 (CONH). .tau. (DMSO-d.sub.6) 2.2-2.9 (thienyl); 6.08 (OMe); 0.13 (NH); 4.12, 4.76 (.beta.-lactam); 6.28, 6.52 (J= 18 Hz. CH.sub.2 in ring); 4.94, 5.27 (J= 13 Hz. CH.sub. 2 OAc), and 7.94 (OAc).
EXAMPLE 9
(a) t-Butyl 3-Acetoxymethyl-7.beta.-(2-t-butoxyimino-2-phenylacetamido)-ceph-3-em-4-ca rboxylate(syn-isomer) was prepared by Method B and purified by chromatography on silica, Method C to give a pale yellow foam (43%) which was crystallised from isopropyl ether to give the ester, m.p. 150.degree.-153.degree., .tau. (CDCl.sub.3) 2.2-2.75 (multiplet, aromatic protons), 2.83 (doublet, NH), 8.61 (t-butoximine), 8.46 (t-butyl ester). Deprotection by Method F gave
(b) 3-Acetoxymethyl-7.beta.-(2-t-butoxyimino-2-phenylacetamido)ceph-3-em-4-car boxylic Acid(syn-isomer) as a pale yellow foam, .nu..sub.max. (CHBr.sub.3) 1782 (.beta.-lactam) 1685, 1522 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) 2.2-2.75 (multiplet, aromatic protons), 0.4 (doublet, NH), 8.70 (t-butoxime).
EXAMPLE 10
(a) t-Butyl 3-Acetoxymethyl- 7.beta.-[2-methoxymethoxyimino-2-(thien-2-yl)acetamido]-ceph-3-em-4-carbox ylate(syn-isomer) .lambda..sub.max. (EtOH) 260 nm (.epsilon.12,940), .nu..sub.max. (CHBr.sub.3) 1778 (.beta.-lactam), 1688 and 1510 cm..sup..sup.-1 (CONH), .tau. (CDCl.sub.3) 2.74 (doublet, NH), 4.76 (--OCH.sub. 2 OCH.sub.3), 6.49 (OCH.sub.2 OCH.sub. 3), 7.91 (OCOCH.sub.3), 8.44 (Bu.sup.t), was prepared by Method A and then treated with trifluoroacetic acid by Method E to give:
(b) 3-Acetoxymethyl-7.beta.-[2-methoxymethoxyimino-2-(thien-2-yl)-acetamido]-c eph-3-em-4-carboxylic Acid(syn-isomer) [.alpha.].sub. D +51.degree. (c 0.8, DMSO), .lambda..sub.max. (pH 6 phosphate buffer) 262.5 nm (.epsilon.16,400), .nu..sub.max. (Nujol) 1778 (.beta.-lactam), 1670, 1530 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) 0.11 (doublet; NH), 4.89 (OCH.sub. 2 OCH.sub.3), 6.62 (OCH.sub.2 OCH.sub. 3), 7.96 (OCOCH.sub.3).
EXAMPLE 11
(a) Diphenylmethyl 3-Cyclopropylcarbonyloxymethyl-7.beta.-(2-ethoxyimino-2-phenylacetamido)ce ph-3-em-4-carboxylate(syn-isomer) .lambda..sub.max. (EtOH) 256.5 nm (.epsilon. 15,800) .nu..sub.max. (CHBr.sub.3) 1780 (.beta.-lactam), 1680, 1510 (CONH), 1720 cm..sup..sup.-1 (esters), .tau. (DMSO-d.sub.6) values include 0.19 (doublet, NH), 3.01 (CHPh.sub.2), 5.76 (CH.sub.2 CH.sub.3), 8.68 (CH.sub.3), 8.35 ##STR72## 9.2 (CH.sub.2 groups in cyclopropane ring) was prepared by Method C and then deprotected by Method F to give
(b) 3-Cyclopropylcarbonyloxymethyl-7.beta.-(2-ethoxyimino-2-phenylacetamido)ce ph-3-em-4-carboxylic Acid(syn-isomer), [.alpha.].sub. D +75.degree., (c 0.86 dioxan) .lambda..sub.max. (pH 6 phosphate buffer) 257 nm (.epsilon. 16,000), .nu..sub.max. (Nujol) 1780 cm..sup..sup.-1 (.beta.-lactam) .tau.(DMSO-d.sub.6) values include 0.25 (doublet NH), 5.80 (CH.sub.2 CH.sub.3), 8.73 (CH.sub.3).
EXAMPLE 12
3-Acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3 -em-4-carboxylic acid(syn-isomer).
To a cold (0.degree.-5.degree.) solution of 3-acetoxymethyl-7.beta.-aminoceph-3-em-4-carboxylic acid (0.565g.) in acetone (10ml) and water (10 ml) containing anhydrous sodium bicarbonate (0.42 g) was added a solution of syn-2 -methoxyiminophenylacetyl chloride (0.495 g.) in acetone (5 ml) over a period of 5 minutes. The solution was stirred at room temperature for 30 minutes and then evaporated to remove acetone. The solution was washed with ether (20 ml), and the aqueous phase acidified under ethyl acetate (50 ml) to pH 2 with 2N-hydrochloric acid. The mixture was filtered and the layers separated; the aqueous phase was extracted with ethyl acetate. The organic extracts were combined, dried and evaporated to give a white solid (0.48 g.). This was triturated with ether, producing the title compound (0.45 g. 64%), [.alpha.].sub.D.sup.17 + 56.degree. (1.04 in dioxan), .lambda..sub.max. (pH 6 phosphate buffer) 259 nm (.epsilon. 19,700 ), .nu..sub.max. (Nujol) 1778 (.beta.-lactam), 1740, 1262 (OCOCH.sub.3) 1712 (CO.sub.2 H), 1660, 1534 cm..sup.-1 (CONH): .tau.(DMSO-d.sub.6) values include 0.2 (doublet, J 8.0 Hz; NH), 2.3-2.6 (multiplet; Ph), 6.05 (singlet; OCH.sub.3), 7.94 (singlet, acetate).
EXAMPLE 13
3-Acetoxymethyl-7.beta.-(2-ethoxyimino-2-phenylacetamido)ceph-3-em-4-carbox ylic acid (syn isomer)
Oxalyl chloride (0.26 ml.) was added to a suspension of sodium syn-ethoxyiminophenylacetate [prepared from the corresponding acid (300 mg.) and 0.43 N sodium methoxide solution (3.62 ml.), and dried over phosphorus pentoxide] in dry benzene (20 ml.) containing one drop of dimethylformamide. After stirring for one hour, the mixture was evaporated to dryness at room temperature. The residue was dissolved in acetone (15 ml.), and this solution was added dropwise to an ice-cooled, stirred solution of 7.beta.-aminocephalosporanic acid (424 mg.), in water (20 ml.) containing sodium bicarbonate (262 mg.). The mixture was stirred for 2 hr., then the acetone was evaporated and the aqueous solution was acidified to pH 1.5. This mixture was extracted with ethyl acetate, the extracts were combined, washed (water, brine) and dried. Evaporation of the extracts gave the title compound (622 mg., 89%) as a colourless foam, [.alpha.].sub.D + 56.degree. (c 1.04 dioxan), .lambda..sub.max. (pH 6 phosphate buffer), 257.5 nm (.epsilon. 20,200), .nu..sub.max. (Nujol) 1772 (.beta.-lactam), 1656 and 1518 cm..sup..sup.-1 (CONH), .tau. values (DMSO-d.sub.6) include 0.23 (doublet, J 8 Hz, NH), 2.3-2.6 (multiplet, aromatic protons), 5.75 (quartet, CH.sub.2 CH.sub.3), 8.68 (triplet, CH.sub.2 CH.sub.3). 7.96 (OCOCH.sub.3).
EXAMPLE 14
3-Acetoxymethyl-7.beta.-(2-t-butoxyiminothien-2'-ylacetamido)ceph-3-em-4-ca rboxylic Acid (syn isomer)
Methanolic sodium methoxide (0.53 N, 7.5 ml) was added dropwise to a stirred solution of syn-t-butoxyimino-thien-2-ylacetic acid (0.9 g.) in dry methanol (10 ml.) at 0.degree.- 5.degree.. The mixture was evaporated to remove methanol and dried by azeotroping with dry benzene. Oxalyl chloride (0.63 ml., 0.95 g.) was added to a stirred suspension of the sodium salt and dimethylformamide (3 drops) in dry benzene (40 ml.). The mixture was stirred at room temperature for 1 hr. and evaporated to remove benzene. The acid chloride in acetone (10 ml.) was added dropwise to a stirred solution of 7.beta.-aminocephalosporanic acid (1.09 g.) and sodium bicarbonate (0.672 g.) in acetone (20 ml.) and water (20 ml.) at 0.degree.-5.degree. and the mixture was stirred at room temperature for 2 hr. The resulting solution was evaporated to remove acetone and washed with ethyl acetate. The aqueous solution was acidified, in the presence of ethyl acetate, with 2 N hydrochloric acid to pH 1.5 and extracted with ethyl acetate. The combined extracts were washed with water, dried and evaporated to give the title compound (1.15 g., 61%) as a pale yellow foam, [.alpha.].sub.D.sup.23 + 61.degree. (c 1.0, dioxan), .lambda..sub.max. (pH 6 phosphate buffer) 260 nm (.epsilon. 12,500), .nu..sub.max. (Nujol) 1780 (.beta.-lactam) 1666 and 1520 cm..sup..sup.-1 (CONH) .tau. values (DMSO-d.sub.6) include 0.28 (doublet, J 8 Hz; NH), 2.36 (1H) and 2.84 (2H) (thien-2-yl protons), 7.98 (OAc), 8.68 (Bu.sup.t).
EXAMPLE 15
3-Acetoxymethyl-7.beta.-(2-t-butoxyiminobenzo[b]-thien-3'-yl acetamido)ceph-3-em-4-carboxylic Acid (syn-isomer)
syn-t-Butoxyiminobenzo-[b]-thien-3-ylacetic acid (0.976 g.) was treated with sodium methoxide solution (0.445 M: 6.1 ml) at room temperature. Evaporation gave the sodium salt as a white powder (1.05 g.). This was dried over phosphorus pentoxide overnight.
The sodium salt (0.5 g.) suspended in dry benzene (10 ml) containing dimethylformamide (1 drop) was treated at room temperature with oxalyl chloride (0.4 ml) for 1 hr. The solvent was removed by evaporation and the residue in acetone (20 ml) was added dropwise to a solution of 7.beta.-aminocephalosporanic acid (0.49 g.) and sodium bicarbonate (0.378 g.) in water (30 ml) at 0.degree.-5.degree.. The resulting pale yellow suspension was stirred at room temperature for 21/2 hr. The acetone was removed by evaporation and the aqueous phase was acidified to pH 2 under ether. The ether fraction was washed with water and dried. Evaporation gave a cream-coloured foam that was dissolved in methylene chloride and re-evaporated to give, after drying, the title compound (0.85 g. 95%). [.alpha.].sub.D + 61.degree. (c 1 dioxan), .lambda..sub.max. (pH 6 buffer) 230, 297, 304 inf. nm (.epsilon. 27,600, 11,500, 10,630), .tau. (DMSO-d.sub.6) values include 0.29 (doublet, NH), 4.04 (double doublet, 7 proton), 4.73 (doublet, 6-proton), 8.59 (Bu.sup.t).
EXAMPLE 16
3-Acetoxymethyl-7.beta.-(2-t-butoxyiminofur-2'-ylacetamido)ceph- 3-em-4-carboxylic Acid (syn-isomer)
syn-t-Butoxyiminofur-2-ylacetic acid (2.11 g.) was treated at room temperature with sodium methoxide (0.525 M: 19.1 ml). The methanol was removed by evaporation and the resulting buff powder was dried over phosphorus pentoxide overnight. The sodium salt (9.33 mg.) in dry benzene (10 ml) containing dimethylformamide (1 drop) was treated with oxalyl chloride (0.8 ml) at room temperature for 1 hr. The solvent was removed by evaporation and the residue in acetone (30 ml) was added dropwise to a solution of 7.beta.-aminocephalosporanic acid (1.09 g) and sodium bicarbonate (672 mg.) in water (50 ml) at 0.degree.-5.degree.. The resulting solution was stirred at room temperature for 11/2 hr. The acetone was removed by evaporation and the aqueous phase was adjusted to pH 8 and washed with a little ether. The aqueous layer was acidified to pH 2.0 under ether and the organic layer was washed with water, dried and evaporated to give a pale yellow foam. This was dried over silica and potassium hydroxide pellets to give the title compound as a pale yellow foam (1.6 g. 86%), [.alpha.].sub.D + 62.degree. (c 1 dioxan). .lambda..sub.max. (pH 6 phosphate buffer) 272 nm (.epsilon. 18,600 ), .nu..sub.max. (CHBr.sub.3) 1780 (.beta.-lactam) 1674 and 1510 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) values include 0.34 (doublet, NH), 2.17 and 3.34 (fur-2-yl, protons) 8.71 (Bu.sup.t).
EXAMPLE 17
3-Azidomethyl-7.beta.-(2-benzyloxyiminophenylacetamido)-ceph-3-em-4-carboxy lic Acid (syn-isomer)
syn-Benzyloxyiminophenylacetic acid (1.28 g.) was treated at room temperature with 0.525 molar sodium methoxide solution (9.55 ml). in methanol. Evaporation gave the sodium salt as a white powder.
The sodium salt, suspended in dry benzene (15 ml.) containing dimethylformamide (2 drops) was treated with oxalyl chloride (0.51 ml.) and kept at room temperature for 1 hr. After filtration through a dry sinter the filtrate was evaporated and the residue in dry methylene chloride (20 ml.) added to a solution of 7.beta.-amino-3-azidomethylceph-3-em-4-carboxylic acid (1.27 g.) and triethylamine (2.07 ml.) in dry methylene chloride (50 ml.) at 0.degree.- 5.degree.. The resulting pale orange solution was stirred at room temperature for 11/4 hr. Evaporation gave a brown foam which was dissolved in water, washed with a little ether and acidified to pH 1.8 under ethyl acetate. The ethyl acetate gave the crude product as a yellow foam (1.9 g., 77%). Precipitation from sodium bicarbonate solution (50 ml.) with dilute acid gave the title compound (1.4 g., 56%). [.alpha.].sub.D + 49.5.degree. (c 1 dioxan) .lambda..sub. max. (pH 6 phosphate buffer) 258.5 nm (.epsilon.19,400) .nu..sub.max. (CHBr.sub.3) 1782 (.beta.-lactam) 1684 and 1512 cm..sup..sup.-1 (CONH). .tau. values (DMSO-d.sub.6) include 0.07 (doublet, J 8 Hz; NH), 2.58 (multiplet, aromatic protons), 4.76 (CH.sub.2 Ph), 4.05 (double doublet, C 7 proton).
EXAMPLE 18
Sodium 3-azidomethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carbox ylate (syn-isomer).
To a cold (0.degree.-5.degree.) solution of 3-azidomethyl-7.beta.-aminoceph-3-em-4-carboxylic acid (4.6 g.) in acetone (100 ml) and water (100 ml) containing sodium bicarbonate (3.66 g.), was added dropwise over 20 minutes a solution of syn 2-methoxyiminophenylacetyl chloride (4.32 g.) and the resulting yellow solution was stirred at room temperature for 30 minutes. Acetone was removed by evaporation, and the dark green solution was washed with ether. The pH of the aqueous phase under ethyl acetate (100 ml) was reduced to 2 with 2 N-hydrochloric acid. The layers were separated, and the aqueous phase was extracted with ethyl acetate. The organic extracts were combined, dried, and evaporated to a pale brown froth (8.2 g.) which was triturated with petroleum spirit (b.p. 60.degree.-80.degree.) and decolourised with charcoal, producing a cream froth (7.67 g.).
This was dissolved in ethyl acetate (30 ml), and a 10% w/v solution of sodium 2-ethyl hexanoate in n-butanol (34.8 ml). was added dropwise to the stirred solution, precipitating the title compound (4.05 g, 51%), [.alpha.].sub.D.sup.23 + 63.5.degree. (c, 1.0 in water), .lambda..sub.max. (pH 6.0 phosphate buffer) 258-259 nm. (.epsilon. 18,850), .nu..sub.max. (Nujol) 2102 (N.sub.3), 1756 (.beta.-lactam), 1596 (CO.sub.2 .sup.-), 1682 and 1528 cm..sup..sup.-1 (CONH); .tau. (DMSO-d.sub.6) values include 0.22 (doublet J 8 Hz; NH), 2.3- 2.6 (multiplet, Ph) and 6.04 (singlet, OCH.sub.3).
EXAMPLE 19-46
General Procedures for the Preparation of 3-Substituted-methyl-7.beta.-(2-substituted oxyimino-2-arylacetamido)-ceph-3-em-4-carboxylic Acids
Method A
A solution of the appropriate syn-2-substituted-oxyimino-2-arylacetyl chloride (prepared from 1 equiv. of the corresponding sodium salt with oxalyl chloride) was dissolved in acetone and the solution was added dropwise to a stirred, ice-cold (0.degree.-5.degree.) solution of 7.beta.-aminocephalosporanic acid or 3-azidomethyl-7.beta.-aminoceph-3-em-4-carboxylic acid (1 equiv.) in water containing sodium bicarbonate (2-2.5 equiv.). The mixture was stirred for 30 min.-2.5 hr. allowing the temperature to rise to room temperature. Acetone was removed by evaporation under reduced pressure, the pH was adjusted to 1.5-2.0 and the product was extracted into ethyl acetate (or occasionally ether). The extracts were washed with water or saturated brine, dried and evaporated to a foam or a solid.
Method B
As in Method A but the product was purified by dissolving in aqueous sodium bicarbonate and reprecipitated by acidification.
Method C
As in Method A but the sodium salt was extracted into ethyl acetate and the extract washed successively with 2 N-hydrochloric acid and water, dried and evaporated to a foam.
Method D
A solution of the appropriate acid chloride (1 equiv.) was dissolved in dry methylene chloride (ca. 5 ml/mmole) and the solution was added to a suspension or a solution of 7.beta.-aminocephalosporanic acid or 3-azidomethyl-7.beta.-aminoceph-3-em-4-carboxylic acid (1 equiv.) and triethylamine (3 equivs.) in methylene chloride at 0.degree.-5.degree.. After stirring at room temperature for 1-1.5 hr. the solution was evaporated to dryness, dissolved in water, washed with ethyl acetate and the aqueous layer acidified to pH 1.5 under ethyl acetate. The ethyl acetate extract was washed with water and saturated brine and evaporated to a foam or solid.
EXAMPLE 47
7.beta.-(2-Benzyloxyiminothien-2'-ylacetamido)-3-(2-methyl-1,3,4 -thiadiazol-5-ylthiomethyl)ceph-3-em-4-carboxylic acid (syn isomer)
Oxalyl chloride (0.45 ml) was added to a stirred suspension of sodium syn-benzyloxyiminothien-2-yl-acetate (0.747 g.) in dry benzene (20 ml.) containing dimethylformamide (one drop). The mixture was stirred for one hour, then evaporated to dryness. A solution of the residue in ethyl acetate (25 ml.) was added dropwise to a stirred suspension of diphenylmethyl 7.beta.-amino-3-(2-methyl-1,3,4-thiadiazol-5-ylthiomethyl)ceph-3-em-4-carb oxylate (1.35 g.) in ethyl acetate (20 ml), containing propylene oxide (1.0 ml.). The solid rapidly dissolved, and the resulting solution was stirred for 16 hr. The solvent was evaporated off, and the residue was redissolved in ethyl acetate (50 ml.). This solution was washed successively with 3% sodium bicarbonate solution, 2 N hydrochloric acid, water, and saturated brine, dried, and evaporated to dryness to give the title compound as its diphenylmethyl ester (2.01 g.), as a pale brown foam. .tau. values include (DMSO-d.sub.6) 0.03 (doublet, NH), 4.79 (CH.sub.2 Ph) 7.26 (CH.sub.3).
To this crude ester (1.67 g.) was added anisole (0.24 g.), and trifluoroacetic acid (10 ml.). The resulting solution was left at room temperature for 10 min. then the trifluoroacetic acid was removed by evaporation at room temperature. The residue was distributed between ethyl acetate and aqueous sodium bicarbonate solution. The aqueous layer was acidified to pH 1.7 then extracted thrice with ethyl acetate. The combined extracts were washed (water, saturated brine), dried, and evaporated to dryness to give a red foam (1.04 g.). The foam was dissolved in aqueous sodium bicarbonate solution, then reprecipitated by addition of 2 N hydrochloric acid. This was repeated to give the title compound (350 mg. 28%) [.alpha.].sub.D - 96.degree. (c 0.97 dioxan), .lambda..sub.max. (pH 6 buffer) 274 nm (.epsilon. 18,500), .nu..sub.max. (Nujol) 1784 (.beta.-lactam) .tau. values include (DMSO-d.sub.6) 0.07 (doublet, NH), 2.60 (Ph), 4.81 (CH.sub.2 Ph) and 7.32 (CH.sub.3).
EXAMPLE 48
(a) t-Butyl 3-acetoxymethyl-7.beta.-(2-methoxyiminopyrid-3'-ylacetamido)-ceph-3-em-4-c arboxylate (syn-isomer)
A suspension of 2-methoxyiminopyrid-3'-ylacetic acid (1.8 g.) in dichloromethane (50 ml.) was cooled to ca. -10.degree. and treated with phosphorus pentachloride (2.08 g.). The mixture was stirred and maintained at ca. -10.degree. for 11/2 hours by which time most of the suspension had dissolved. The cold mixture was treated with t-butyl 7.beta.-amino-3-acetoxymethylceph-3-em-4-carboxylate (3.0 g.) and propylene oxide (2 ml) and stirred for a further 30 minutes. The solution was partitioned between ethyl acetate and saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic extracts were washed with water and then extracted with 2 N hydrochloric acid. Unchanged amine was removed by the addition of an excess of sodium nitrite to the cooled acidic extract followed by washing with ethyl acetate after ca. 5 minutes at 0.degree.. The resulting aqueous phase was neutralised with saturated sodium bicarbonate solution and extracted with ethyl acetate. The organic extract was washed with water, dried over magnesium sulphate and evaporated to give a mixture of the syn- and anti- isomers of the title compound as a foam (1.33 g.).
Chromatography on silica gel using chloroformmethanol (99:1) gave the anti-isomer (394 mg.) followed by the syn-isomer as a foam (683 mg.). The syn-isomer was dissolved in ethyl acetate and precipitated by adding to petroleum spirit bp. 40.degree.-60.degree.. The amorphous solid was collected, washed with petroleum spirit and dried to yield the title ester (484 mg., 10%), [.alpha.].sub.D.sup.20 + 49.degree. (c 0.45, DMSO), .lambda..sub.max. (EtOH) 255 nm (.epsilon. 16,100), .nu..sub.max. (CHBr.sub.3) 3396 (NH), 1786 (.beta.-lactam), 1735 (OAc), 1724 (CO.sub.2 R), 1682 and 1518 cm..sup..sup.-1 (CONH), .tau. (d.sub.6 DMSO) 0.14 (d, J 8 Hz; NH), 1.26, 1.30, 2.03 and 2.48 (multiplets; aromatic protons), 4.10 (q, J 5 and 8 Hz; C-7H), 4.74 (d, J 5 Hz; C-6H), 5.03 and 5.36 (2 d, J 13 Hz; C-3 CH.sub.2), 6.03 (s; OCH.sub.3), 6.28 and 6.52 (2d, J 18 Hz; C-2 H.sub.2), 7.97 (s; OCOCH.sub.3), 8.52 (s; C(CH.sub.3).sub.3).
(b) 3-Acetoxymethyl-7.beta.-(2-methoxyiminopyrid-3'-ylacetamido)-ceph-3-em-4-c arboxylic acid trifluoroacetic acid salt (syn-isomer)
A solution of t-butyl-3-acetoxymethyl-7.beta.-(2-methoxyiminopyrid-3-ylacetamido)-ceph-3 -em-4-carboxylate syn-isomer (440 mg.), in trifluoroacetic acid (7 ml.) was stood at 20.degree. for 15 minutes. The solvent was removed and the residue evaporated three times from benzene. The resulting gum was dissolved in a small volume of acetone and added slowly to an excess of stirred petroleum spirit (bp 40.degree.-60.degree.). The solid was collected by filtration, washed well with petroleum spirit and dried to afford the title syn acid salt as a solvated pale yellow powder (561 mg.), [.alpha.].sub.D.sup.20 +30.degree. (c 0.88, DMSO), .lambda..sub.max. (pH 6 buffer) 254 nm (.epsilon. 14,300), .nu..sub.max. (Nujol) 3250 (NH), 2600 and 1720 (CO.sub.2 H), 1782 (.beta.-lactam), 1736 (OAc), 1670 (CF.sub.3 CO.sub.2 .sup.-), 1670 and 1550 cm..sup..sup.-1 (CONH), .tau. (d.sub.6 -DMSO) 0.08 (d, J 8 Hz; NH), 1.16, 1.22, 1.90 and 2.36 (multiplets; aromatic protons), 4.06 (q, J 5 and 8 Hz; C-7H), 4.73 (d, J 5 Hz; C-6H), 4.93 and 5.26 (2d, J 13 Hz; C-3 CH.sub.2), 5.98 (s; OCH.sub.3), 6.26 and 6.49 (2d, J 18 Hz; C-2 H.sub.2), 7.94 (s; OCOCH.sub.3).
EXAMPLE 49
7.beta.-(2-Methoxyimino-2-phenylacetamido)-3-pyridiniummethyl-ceph-3-em-4-c arboxylate (syn-isomer).
A solution of 3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carb oxylic acid (syn-isomer) (5 g.) in water (30 ml), acetone (30 ml) and pyridine (2.8 ml.) was heated at 50.degree. for 4 hours. More pyridine (2.8 ml) was added, and the solution (pH .about. 5) was heated at 50.degree. for 16 hours. The dark brown solution was decanted from a gum, evaporated to remove acetone, and washed with methylene chloride. Excess solvent was removed by evaporation, and the solution passed down a column of Dowex 1 ion exchange resin in the acetate cycle and the column eluted with water. The eluate was freeze dried, and the solid was triturated with acetone, producing the title compound as a cream solid (0.84 g), [.alpha.].sub.D.sup.23 - 6.4.degree. (c, 0.94 in water), .lambda..sub.max. (pH 6.0 phosphate buffer) 257 nm (.epsilon. 19,900), .nu..sub.max. (Nujol) 1770 (.beta.-lactam), 1660, 1540 (CONH) and 1604 cm..sup..sup.-1 (CO.sub.2.sup.-); .tau. (D.sub.2 O) values include 1.03, 1.90 and 1.40 (.alpha.,.beta. and .gamma.-protons in the pyridinium ring), 2.2-2.6 (multiplet, Ph), 5.99 (singlet, OCH.sub.3), .tau. (DMSO-d.sub.6) values include 0.27 (doublet, J 8 Hz; NH), 6.10 (singlet, OCH.sub.3).
EXAMPLE 50
7.beta.-(2-Methoxyimino-2-phenylacetamido)- 3-(4-carbamoylpyridiniummethyl)-ceph-3-em-4-carboxylate (syn-isomer)
3-Acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)-ceph-3-em-4-carb oxylic acid (syn isomer) (5 g) and isonicotinamide (4.2 g) were mixed in water (150 ml) and acetone (30 ml.) and the suspension heated at 50.degree. for 20 hours. The mixture was cooled, filtered, evaporated to remove acetone, and the aqueous solution was passed down a column (3.times. 16 cm.) of Dowex 1 ion exchange resin in the acetate cycle. After washing the column with water the eluate was freeze dried, and the resulting solid was stirred with acetone (250 ml.) for 30 minutes, and the suspension filtered, producing the title compound (1.4 g.), [.alpha.].sub.D.sup.23 - 50.degree. C. (c 1.06 in DMSO), .lambda..sub.max. (pH 6 buffer) 260 nm. (.epsilon. 20,900), .nu..sub.max.sup.Nujol 1770 (.beta.-lactam), 1675 and 1560 (CONH) and 1605 cm..sup..sup.-1 (CO.sub.2.sup.-), .tau. (DMSO-d.sub.6) values include 0.33, 1.49 (.alpha., and .beta. protons in pyridinium ring), 1.19 and 1.74 (--CONH.sub.2), 2.4-2.6 (multiplet, Ph), 0.26 (J, 8 Hz; NH), 6.10 (singlet; OCH.sub.3).
EXAMPLE 51
Sodium 3-hydroxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em- 4-carboxylate (syn-isomer).
3-Acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carbo xylic acid (syn-isomer) (2.0 g.) was dissolved in the minimum volume of saturated sodium bicarbonate solution and the solution was diluted with water (160 ml) and warmed to 37.degree.. Defatted wheat-gram (15 g) was added and the mixture was stirred for 31/2 hours during which time the pH remained at 7.4. The mixture was then poured into acetone (200 ml) and filtered through kieselguhr. The acetone was evaporated off under reduced pressure at 30.degree. and the aqueous solution was covered with ethyl acetate, taken to pH 2.7 by the addition of concentrated hydrochloric acid. The layers were separated and the aqueous layer was reextracted with ethyl acetate. The combined ethyl acetate extracts were washed with water. More water was added to the ethyl acetate solution and the pH was taken to 7 by careful addition of 2 N sodium hydroxide solution. The organic layer was further extracted with water and the combined aqueous solution was washed with ethyl acetate and ether, degassed and lyophilised to give the title methoxime as a brown solid (1.52 g; 82%) [.alpha.].sub.D + 104.degree. (c 1.1, DMSO) .lambda..sub.max. (pH 6 phosphate) 258.5 nm (.epsilon. 14,100) .nu..sub.max. (Nujol) 1730 (.beta.-lactam), 1570 (COO.sup.- ), 1640 and 1520 cm.sup..sup.-1 (CONH), .tau. (D.sub.2 O; 100 MHz) 2.2 to 2.6 (multiplet, aromatic protons), 4.16 (doublet, J 4 Hz; C-7H), 4.79 (doublet, J 4 Hz; C-6H), 5.72 (singlet; C-2 CH.sub.2), 5.97 (singlet; NOCH.sub.3), 6.32 and 6.59 [ 2 doublets (branches of quartet), J 18 Hz; C-3 CH.sub.2).
EXAMPLE 52
3-Hydroxymethyl-7.beta.-(2-benzyloxyiminothien-2'-ylacetamido)-ceph-3-em-4- carboxylic Acid (syn isomer)
To a suspension of 3-acetoxymethyl-7.beta.-(2-benzyloxyiminothien-2'-ylacetamido)ceph-3-em-4- carboxylic acid (1.0 g.) in water (50 ml.) was added sufficient saturated sodium bicarbonate solution to dissolve the solid, and bring the solution to pH 7.0. Defatted wheat germ (8.0 g.) was added to the solution, and the mixture was stirred for 21 hr., keeping the pH at 6.8 to 7.0 by adding acetic acid. After 5 hr., the pH did not alter, and TLC (CHCl.sub.3 :MeOH:AcOH; 18:2:1) on an acidified aliquot showed reaction to be almost complete.
The mixture was filtered, and the pH of the filtrate was adjusted to pH 4.5. Filtration through keiselguhr gave a yellow solution, whose pH was adjusted to 2.5 under ethyl acetate. The aqueous mixture was extracted with ethyl acetate, and the combined extracts were washed (water, brine), dried, and evaporated to dryness to give a brown solid. Trituration of this solid with methylene chloride gave a colourless solid (480 mg.), whose n.m.r. spectrum was consistent with that expected of the hydroxymethyl compound.
A portion of the solid (366 mg.) dissolved in 50% aqueous ethanol (10 ml.) was carefully neutralised with 0.1 N sodium hydroxide solution (3.02 ml., required). The ethanol was evaporated, and the aqueous mixture was extracted thrice with water. Evaporation of the aqueous solution gave sodium 3-hydroxymethyl-7.beta.-(2-benzyloxyiminothien-2'-ylacetamido)ceph-3-em-4- carboxylate dihydrate (132 mg.), [.alpha.].sub.D + 88.degree. (c, 0.89, DMSO), .lambda..sub.max. (pH 6 phosphate buffer) 262 nm (.epsilon. 16,100) .nu..sub.max. (Nujol) 1744 (.beta.-lactam), 1650 and 1522 (CONH), 1580 (CO.sub.2.sup.-) cm..sup..sup.-1, .tau. (DMSO-d.sub.6) values include 0.13 (doublet, NH), 2.54 (Ph) 4.76 (CH.sub.2 Ph), 4.26 (quartet, 7-proton).
EXAMPLE 53
3-(2-Chloroethylcarbamoyloxymethyl)-7.beta.-(2-methoxyiminophenylacetamido) ceph-3-em-4-carboxylic acid (syn-isomer)
A solution of 3-hydroxymethyl-7.beta.-(2-methoxyiminophenylacetamido)ceph-3-em-4-carboxy lic acid (syn-isomer) (0.35 g.), 2-chloroethylisocyanate (0.097 g.) and triethylamine (1.79 g.) in dimethylformamide (10 ml.) was stirred at room temperature for two hours. The reaction mixture was poured into saturated sodium bicarbonate solution, and the aqueous phase washed with ethyl acetate, acidified and extracted with ethyl acetate. The combined extracts were dried, concentrated to ca. 5 ml. and added dropwise with stirring to petroleum spirit (250 ml.). The resulting solid was filtered and dried to give the title acid (0.157 g., 35%) .lambda..sub.max. (pH 6 buffer) 258 nm (.epsilon. 16,800), .nu..sub.max. (Nujol) 3250 (NH), 2600 and 1715 (CO.sub.2 H), 1700 and 1530 (NHCO.sub.2 R), 1660 and 1540 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) values includes 0.22 (doublet J 9 Hz; 7.beta.-NH), 2.3 to 2.6 (multiplet; aromatic protons), 6.05 (singlet; OCH.sub.3), 6.40 (multiplet; C-2 CH.sub.2 and --CH.sub.2 Cl), 6.65 (multiplet; --NHCH.sub.2).
EXAMPLE 54
(a) t-Butyl-3-acetoxymethyl-7.beta.-(pyrid-4-ylacetamido)ceph-3-em-4-carboxyla te
A suspension of pyrid-4-ylacetyl chloride hydrochloride (6.0 g.) in ethyl acetate (50 ml) containing propylene oxide (12 ml.) was stirred, cooled in ice and treated dropwise with a solution of t-butyl 3-acetoxymethyl-7.beta.-aminoceph-3-em-4-carboxylate (5.0 g.) in ethyl acetate (50 ml.). The mixture was stirred at 20.degree. for 20 hours and then washed with saturated sodium bicarbonate solution and extracted with 2 N hydrochloric acid. The aqueous extract was washed with ethyl acetate, neutralised with saturated sodium bicarbonate solution and extracted with ethyl acetate. The extract was washed with water, dried over magnesium sulphate and evaporated to small bulk whereupon crystallisation began. The crystalline solid was washed well with petroleum spirit (bp. 40.degree.-60.degree.) and dried to give the title ester as a buff coloured powder (5.4 g., 80% based on amine). A portion (250 mg.) was recrystallised from ethyl acetate as white needles (170 mg.), [.alpha.].sub.D +94.degree. (c 0.9, DMSO) .lambda..sub.max. (EtOH) 256.5 nm (.epsilon. 9,900), .nu..sub.max. (CHBr.sub.3) 3412 (NH), 1784 (.beta.-lactam), 1736 (OAc), 1722 (CO.sub.2 R), 1690 and 1512 cm..sup..sup.-1 (CONH), .tau. (d.sub.6 -DMSO) 0.81 (d, J 8 Hz; NH), 1.47 and 2.68 (2d, J 5 Hz; aromatic protons), 4.27 (q, J 8 and 5 Hz; C-7H), 4.85 (d, J 5 Hz; C-6 H), 5.01 and 5.36 (2d, J 13 Hz; C-3CH.sub.2), 6.38 (s; CH.sub.2 CONH), 6.39 (s; C-2H.sub.2), 7.96 (s; OCOCH.sub.3), and 8.50 (s; C(CH.sub.3).sub.3).
(b) t-Butyl 3-acetoxymethyl-7.beta.-(2-hydroxyiminopyrid-4'-ylacetamido)-ceph-3-em-4-c arboxylate
A solution of t-butyl 3-acetoxymethyl-7.beta.-(pyrid-4-ylacetamido)ceph-3-em-4-carboxylate (3.0 g.) in acetic acid (30 ml.) was stirred, cooled briefly in ice and treated dropwise over 3-4 minutes with a solution of sodium nitrite (1.38 g.), in water (10 ml.). The mixture was stirred at 20.degree. for 30 minutes and was then diluted with water and extracted with ethyl acetate. The organic extract was washed with saturated sodium bicarbonate solution and water and then dried over magnesium sulphate. Evaporation almost to dryness gave a residue which was dissolved in the minimum volume of acetone and added dropwise to stirred petroleum spirit (bp. 40.degree.-60.degree.). The resulting solid was collected, washed with petroleum spirit and dried to yield the title oxime as a white powder (2.9 g., 91%) which was shown (NMR) to be a syn-/anti-mixture (30:70).
The oxime mixture (2.5 g) was chromatographed on silica gel and eluted with methanol-chloroform (1:49) to give the title compound (anti-isomer).
Further elution with methanol-chloroform (1:49) gave material which was dissolved in acetone and added to stirred petroleum (bp. 40.degree.-60.degree.) to afford t-butyl 3-acetoxymethyl-7.beta.-(2-hydroxyiminopyrid-4'-ylacetamido)ceph-3-em-4-ca rboxylate (syn-isomer, 0.81 g., 32% from mixture), [.alpha.].sub.D + 57.degree. (c, 1.11, DMSO), .lambda..sub.max. (EtOH) 253.5 nm (.epsilon. 17,600), .nu..sub.max. (Nujol) 3220 (NH), 1784 (.beta.-lactam), 1744 (OAc), 1710 (CO.sub.2 R) 1640 and 1526 cm..sup..sup.-1 (CONH), .tau. (d.sub.6 -DMSO) -2.26 (s; NOH), 0.27 (d, J 8 Hz; NH), 1.33 and 2.47 (2d, aromatic protons), 4.04 (q, J 5 and 8 Hz; C-7H), 4.71 (d, J 5 Hz; C-6H), 4.98 and 5.33 (2d, J 13 Hz; C-3 CH.sub.2), 6.20 and 6.53 (2d, J 18 Hz; C-2 H.sub.2) 7.94 (s, OCOCH.sub.3), 8.49 (s, C(CH.sub.3).sub.3).
(c) t-Butyl 3-acetoxymethyl-7.beta.-(2-methoximinopyrid-4'-ylacetamido)-ceph-3-em-4-ca rboxylate (syn-isomer)
A solution of t-butyl 3-acetoxymethyl-7.beta.-(2-hydroxyiminopyrid-4'-ylacetamido)-ceph-3-em-4-c arboxylate (syn-isomer) (50 mg.), in tetrahydrofuran (10 ml.) was cooled to .degree. and treated with an excess of ethereal diazomethane. The mixture was stirred at ca. 5.degree. for 45 minutes and then washed with water. The organic extract was dried and evaporated to yield the title ester as a yellow foam (48 mg., 93%), [.alpha.].sub.D + 55.5.degree. (c 0.88, DMSO), .lambda..sub.max. (EtOH) 257.5 nm (.epsilon. 16,580), .nu..sub.max. (CHBr.sub.3) 3390 (NH) 1784 (.beta.-lactam), 1736 (OAc), 1726 (CO.sub.2 R), 1684 and d1516 cm..sup..sup.-1 (CONH), .tau. (d.sub.6 -DMSO) 0.20 (d, J 8 Hz; NH), 1.33 and 2.48 (multiplets; aromatic protons), 4.09 (q, J 5 and 8 Hz; C-7H), 4.74 (d, J 5 Hz; C-6H), 5.01 and 5.35 (2d, J 13 Hz; C-3CH .sub.2), 5.99 (s, NOCH.sub.3), 6.38 (broad singlet; C-2H.sub.2), 7.96 (s; OCOCH.sub.3), 8.52 (s; C(CH.sub.3).sub.3).
(d) 3-Acetoxymethyl-7.beta.-(2-methoxyiminopyrid-4'-ylacetamido)-ceph-3-em-4-c arboxylic acid trifluoroacetic acid salt (syn-isomer)
A solution of t-butyl 3-acetoxymethyl-7.beta.-(2-methoxyiminopyrid-4'-ylacetamido)-ceph-3-em-4-c arboxylate (syn-isomer) (0.35 g.) in trifluoroacetic acid (10 ml) was stood at 20.degree. for 15 minutes. The solution was diluted with benzene and evaporated to dryness to give a gum which was tritutated with ether to afford the title acid salt as a solid (0.26 g., 67%), [.alpha.].sub.D + 42.degree. (c 0.45, DMSO), .lambda..sub.max. (pH 6 buffer) 255 nm (.epsilon. 19,000), .nu..sub.max. (Nujol) 3250 (NH), 2600 and 1720 (CO.sub.2 H), 1778 (.beta.-lactam), 1734 (OAc), 1666 (CF.sub.3 CO.sub.2.sup.-), 1670 and 1520 cm..sup..sup.-1 (CONH), .tau. (DMSO-d.sub.6) 0.10 (d, J 8 Hz; NH), 1.23 and 2.36 (multiplets; aromatic protons), 4.11 (q, J 5 and 8 Hz; C-7H), 4.76 (d, J 5 Hz; C-6H), 4.98 and 5.29 (2d, J 13 Hz; C-3 CH.sub.2), 5.97 (s; NOCH.sub.3), 6.29 and 6.52 (2d, J 18 Hz; C-2H.sub.2), 7.98 (s; OCOCH.sub.3).
EXAMPLE 55
3-Acetoxymethyl-7.beta.-[2-aminoethoxyimino(thien-2-yl)acetamido]ceph-3-em- 4-carboxylic acid, trifluoroacetate salt (syn isomer)
A solution of syn-(2-t-butoxycarboxamido)ethoxyiminothien-2-ylacetyl chloride (from the acid 314 mg) dissolved in acetone (10 ml.), was added dropwise to an ice-cooled, stirred solution of 7.beta.-aminocephalosporanic acid (272 mg.) in water (20 ml.) containing sodium bicarbonate (168 mg.). The solution was stirred for two hours, then the acetone was evaporated off. The aqueous mixture was acidified to pH 2.0 under ethyl acetate, and extracted with ethyl acetate. The combined extracts were washed (water, brine), dried, and evaporated to dryness to give 3-acetoxymethyl-7.beta.-[2-t-butoxy-carboxamido ethoxyimino(thien-2-yl)acetamido]ceph-3-em-4-carboxylic acid (540 mg., 95%), .tau. (DMSO-d.sub.6) values include 0.27 (d, NH), 7.96(s, OCOCH.sub.3), 8.62 (s, C(CH.sub.3).sub.3).
This crude acid (500 mg.), was dissolved in trifluoroacetic acid (5 ml.), and the solution was stood at room temperature for 5 min. and then was evaporated to dryness at room temperature. Dry benzene (25 ml.) was added to the residue, and the mixture was re-evaporated to dryness. The oily residue was triturated with ether to give the title compound (346 mg., 64%) as a colourless solid [.alpha.].sub.D + 39.degree. (c 0.97 dioxan), .lambda..sub.max. (pH 6 phosphate buffer) 283 nm (.epsilon. 16,500), .nu..sub.max. (Nujol) 1780 cm..sup.-.sup.1, .tau. (DMSO-d.sub.6) values include 0.31 (doublet, J 8 Hz; NH), 1.98 (N.sup.+H.sub.3), 4.08 (C-7 proton), 5.66 (--OCH.sub.2), 6.8 (--CH.sub.2 --N<), 7.96 (OCOCH.sub.3).
EXAMPLE 56
7.beta.-[2-t-butoxyimino-2-(benzo-[b]-thien-3-yl)-acetamido]-3 -(5-methyl-1,3,4-thiadiazol-2-yl)thiomethylceph-3-em-4-carboxylic acid (syn-isomer)
A solution of 2-(t-butoxyimino)-2-(benzo-[b]-thien-3-yl)-acetic acid (1.11 g) in methylene chloride (5 ml.) and dimethylformamide (1 ml) was added to a solution of diphenylmethyl 7.beta.-amino-3-(5-methyl-1,3,4-thiadiazol-2-yl)thiomethylceph-3-em-4-carb oxylate (2.04 g) in methylene chloride (20 ml.). A solution of dicyclohexylcarbodiimide (0.823 g) in methylene chloride (5 ml) was added to the above stirred solution at 20.degree.. The mixture was kept at 5.degree. for 16 hours and was then filtered. The filtrate was washed with 2 N hydrochloric acid, dilute sodium bicarbonate solution, water and brine, dried (MgSO.sub.4) and concentrated under reduced pressure to give the crude diphenylmethyl ester of the title compound as an orange foam (3.37 g.). The foam was dissolved in anisole (6 ml) and the solution was treated with trifluoroacetic acid (25 ml.). After 5 minutes at 20.degree. the solution was concentrated under reduced pressure. The residue was dissolved in ethyl acetate and the resulting solution was again concentrated. The residue was then partitioned between ethyl acetate and dilute sodium bicarbonate solution. The ethyl acetate layer was thoroughly extracted with dilute sodium bicarbonate solution and the combined extracts were washed with ethyl acetate and then were covered with ethyl acetate and taken to pH 2 with concentrated hydrochloric acid. The layers were separated and the aqueous layer was extracted with ethyl acetate. The combined ethyl acetate extracts were washed with water and brine, dried and concentrated under reduced pressure to a low volume. The solution was then added dropwise to stirred petroleum (bp. 40.degree.-60.degree., 400 ml) and the precipitate was collected and washed with petroleum to give the title acid as a buff solid (1.0 g, 41%), [.alpha.].sub.D -87.degree.(c 1 in DMSO) .lambda..sub.max. (pH 6 phosphate buffer) 277.5 nm (.epsilon. 16,850) .nu..sub.max. (Nujol) include 1786 (.beta.-lactam), 1672 and 1520 cm..sup.-.sup.1 (CONH) .tau. values (DMSO-d.sub.6) include 0.27 (d, J 8 Hz; NH), 1.38 (m; H-4 benzo-[b]-thienyl) 1.86 (m; benzo-[b]-thienyl H-7), 2.15 (s; benzo-[b]-thienyl H-2) 2.44 (m; benzo-[b]-thienyl H-5 and H-6), 4.05 (dd, J 8 and 5 Hz; H-7), 4.72 (d, J 5 Hz; H-6) 5.40+ 5.74 (2ds, J 13 Hz; C-3 CH.sub.2), 6.13+ 6.37 (2 ds, J 18 Hz; C-2 CH.sub.2), 7.29 (s; CH.sub.3) and 8.57 (s; C(CH.sub.3).sub.3).
EXAMPLE 57
7.beta.-[2-Ethoxyimino-2-phenylacetamido]-3-(5-methyl-1,3-4-thiadiazol-2-yl )thiomethylceph-3-em-4-carboxylic acid (syn-isomer),
was prepared essentially as described in Example 56 (61% yield) [.alpha.].sub.D - 86.degree. (c 1.1 in DMSO) .lambda..sub.max. (pH 6 phosphate) 262.5 nm (.epsilon. 22,350) .nu..sub.max. (Nujol) include 1780 (.beta.-lactam) and 1664 and 1520 cm..sup.-.sup.1 (CONH), .tau. values (DMSO-d.sub.6) 0.21 (d, J 8 Hz; NH), 2.46 (m; aromatic protons) 4.06 (dd, J 8 and 5 Hz; H-7), 4.73 (d, J 5 Hz; H-6), 5.40+ 5.72 (2ds, J 14 Hz; C-3 CH.sub.2), 5.74 (q, J 7 Hz; CH.sub.3 CH.sub.2 O) 6.11+ 6.39 (2 ds, J 18 Hz; C-2 CH.sub.2), 7.27 (s; CH.sub.3 in thiadiazole) 8.68 (t, J 7 Hz; CH.sub.3 CH.sub.2 O).
EXAMPLE 58
7.beta.-[2-t-Butoxyimino-2-(thien-2-yl)acetamido]-3-(5-methyl-1,3,4-thiadia zol-2-yl)thiomethylceph-3-em-4-carboxylic acid (syn-isomer) was prepared as in Example 56 to give the acid (58% yield) [.alpha.].sub.D - 76.degree. (c 1, DMSO) .lambda..sub.max. (pH 6 phosphate) 271 nm (.epsilon. 14,400) .nu..sub.max. (Nujol) include 1780 (.beta.-lactam) and 1670 and 1530 cm..sup.-.sup.1 (CONH), .tau. values (DMSO-d.sub.6) include 0.26 (d, J 8 Hz; NH), 2.32+ 2.80 (ms; thienyl protons), 4.08 (dd, J 8 and 5 Hz; H-7), 4.73 (d, J 5 Hz; H-6) 5.38+ 5.73 (2 ds, J 13 Hz; C-3 CH.sub.2), 6.10+ 6.36 (2 ds, J 18 Hz; C-2 CH.sub.2), 7.27 (s; CH.sub. 3) and 8.64 (s, C(CH.sub.3).sub.3).
EXAMPLE 59
(a) t-Butyl-3-acetoxymethyl-7.beta.-(2-ethoxyiminopyrid-4-ylacetamido)-ceph-3- em-4-carboxylate (syn-isomer)
A solution of t-butyl-3-acetoxymethyl-7.beta.-(2-hydroxyiminopyrid-4-ylacetamido)-ceph-3 -em-4-carboxylate (syn-isomer) (1.15 g) in tetrahydrofuran (20 ml) was treated with an excess of diazoethane at 20.degree. for 40 minutes. The mixture was diluted with ether, washed with water and extracted with 2 N-hydrochloric acid. The aqueous extract was neutralised with saturated sodium bicarbonate solution and extracted with ethyl acetate. Evaporation of the dried organic extract offered the title ester as a foam (0.84 g, 69%), [.alpha.].sub.D + 47.degree. (c 0.84, DMSO), .lambda..sub.max. (EtOH) 259 nm (.epsilon. 15,450), .nu..sub.max. (Nujol) 3300 (NH), 1788 (.beta.-lactam), 1740 (OAc), 1724 (CO.sub.2 R), 1678 and 1540 cm..sup.-.sup.1 (CONH), .tau. (d.sub.6 -DMSO) values include 0.12 (d, J 8 Hz; NH), 1.26 and 2.42 (multiplets; aromatic protons), 5.69 (q, J 7 Hz, OCH.sub.2 CH.sub. 3), 7.93 (s; OCOCH.sub.3), 8.50 (s; (CH.sub.3).sub.3) and 8.69 (t, J 7 Hz; OCH.sub.2 CH.sub.3).
(b) 3-Acetoxymethyl-7.beta.-(2-ethoxyiminopyrid-4-ylacetamido)-ceph-3-em-4-car boxylic acid trifuoroacetic acid salt (syn-isomer)
t-Butyl-3-acetoxymethyl-7.beta.-(2-ethoxyiminopyrid-4-ylacetamido)-ceph-3-e m-4-carboxylate (0.75 g) was deprotected with trifluoroacetic acid (10 ml) in the usual way to afford the title salt as a powder (0.61 g, 73%), [.alpha.].sub.D + 30.degree. (c 0.4, DMSO), .lambda..sub.max. (pH6 buffer) 257 nm (.epsilon. 12,800), .nu..sub.max. (Nujol) 3300 (NH), 2600 and 1720 (CO.sub.2 H), 1782 (.beta.-lactam), 1740 (OAc), 1680 (CF.sub.3 CO.sub.2 .sup.-), 1670 and 1552 cm..sup.-.sup.1 (CONH), .tau. (d.sub.6 -DMSO) values include 0.06 (d, J 8 Hz; NH), 1.20 and 2.28 (multiplets; aromatic protons), 5.65 (q, J 7 Hz; OCH.sub.2 CH.sub.3), 7.92 (s; OCOCH.sub.3) and 8.64 (t. J 7 Hz; OCH.sub.2 CH.sub.3).
EXAMPLES 60-65
The general procedures for the preparation of 3-substituted methyl-7.beta.-(2-substituted oxyimino-2-arylacetamido)ceph-3-em-4-carboxylic acids described in Examples 19-46 were employed to prepare the compounds listed in tabular form below. (the Table may be regarded as a continuation of Table 2).
EXAMPLE 66
Sodium 3-Acetoxymethyl-7.beta.-(2-phenoxyimino-2-phenylacetamido)-ceph-3-em-4-car boxylate (syn isomer)
A solution of syn-2-phenoxyimino-2-phenylacetic acid (8.6 g) in dry methanol (30 ml) was neutralised with 1.105 N sodium methoxide solution in methanol 32.3 ml). The methanol was evaporated, and the residual sodium salt (dried 48 hr. in vacuo over phosphorus pentoxide) was suspended in dry benzene (75 ml) containing one drop of N,N-dimethylformamide. This suspension was treated with oxalyl chloride (4.5 ml), the mixture was stirred for one hour, and then the solvent and excess reagent were evaporated. A solution of the residual acid chloride in acetone (250 ml) was added over 30 min. to a stirred, ice-cooled solution of 7.beta.-aminocephalosporanic acid (9.71 g) in water (500 ml) and acetone (250 ml), containing sodium bicarbonate (6.5 g). When addition was complete, the mixture was stirred for a further two hours, then the acetone was evaporated under reduced pressure. The aqueous mixture was covered with ether (500 ml) and acidified to pH 1.8; the acidic layer was further extracted with ether. The combined ether extracts were washed (water, saturated brine), dried and evaporated to dryness to give syn-3-acetoxymethyl-7.beta.-(2-phenoxyimino-2-phenylacetamido)ceph-3-em-4- carboxylic acid as a pale yellow foam (15.06 g).
To a stirred solution of the above acid (14.5 g) in acetone (150 ml) was added (15 min) a solution of sodium 2-ethylhexanoate (5.5 g) in acetone (50 ml). The precipitated solid was collected, washed with acetone, and dried in vacuo to give the title compound (12.92 g); [.alpha.].sub.D.sup.22 + 111.degree. (c 1.13, DMSO); .lambda..sub.max. (pH 6 buffer) 260 nm (.epsilon. 19,400), .lambda. infl. 285 nm (.epsilon. 12,000); .nu..sub.max. (Nujol) 3270 (NH), 1752 cm..sup.-.sup.1 (.beta.-lactam); .tau. values (DMSO-d6) include -0.05 (d) (NH), 4.16 (dd) (7-H), 4.82 (d) (6-H), 7.97 (s)(-O.COCH.sub.3).
EXAMPLES 67-76
General Procedures for the Preparation of 3-Acetoxymethyl-7.beta.-(2-substituted oxyimino-2-arylacetamido)-ceph-3-em-4-carboxylic Acids
Method A
A solution of the appropriate syn-2-substituted oxyimino-2-arylacetyl chloride (prepared from 1 equiv. of the corresponding sodium salt with oxalyl chloride) was dissolved in acetone and the solution was added dropwise to a stirred, cold (0.degree.-5.degree.) solution of 7.beta.-aminocephalosporanic acid (1 equiv.) in water containing sodium bicarbonate (2-2.5 equiv.). The mixture was stirred for 0.5-2.5 hr. allowing the temperature to rise to room temperature. Acetone was removed by evaporation under reduced pressure, ethyl acetate was added, the pH was adjusted to 1.5-2.0 and the product was extracted into ethyl acetate. The extracts were washed with water and saturated brine, dried and evaporated to a foam or solid.
Method B
As in Method A but the product was extracted into ether.
Cephalosporin derivatives so prepared are listed in Table 5.
EXAMPLE 77
3-(1-Methyltetrazol-5-ylthiomethyl)-7.beta.-[2-phenoxyimino-2-phenylacetami do]-ceph-3-em-4-carboxylic acid (syn-isomer)
A suspension of 2-phenoxyimino-2-phenylacetic acid (464 mg) in dry methylene dichloride (25 ml) was added, over 5 minutes, to a stirred solution of diphenylmethyl 7.beta.-amino-3-(1-methyltetrazol-5-ylthiomethyl)-ceph-3-em-4-carboxylate (989 mg) and dicyclohexylcarbodiimide (454 mg) in methylene dichloride (25 ml). After stirring for a further 2 hours, the solvent was evaporated in vacuo; the residue was suspended in ethyl acetate (60 ml), the insoluble material filtered off, and the filtrate washed with saturated sodium bicarbonate:water= 1:1, water, and brine (30 ml of each), dried, and evaporated to a foam (1.56 g). A solution of this foam in benzene (10 ml) was purified by chromatography on kieselgel (60 g), with benzene:ethyl acetate= 10:1 as eluant. Appropriate fractions were combined and evaporated to dryness in vacuo to give diphenylmethyl 3-(1-methyltetrazol-5-ylthiomethyl)-7.beta.-[2-phenoxyimino-2-phenylacetam ido]-ceph-3-em-4-carboxylate (syn-isomer) as an orange foam (1.16 g, 87%). A solution of this foam in a mixture of trifluoroacetic acid (4ml) and anisole (1ml) was kept at 23.degree. for 5 minutes, and the solvents were removed at 40.degree. (2mm). The residue was suspended in ether (80 ml), saturated sodium bicarbonate:water= 1:3 (120ml) was added, and the mixture was stirred until all the solid had dissolved (ca. 10 minutes). The aqueous phase was separated, covered with ethyl acetate (150 ml), and acidified to pH 2 with 2N-hydrochloric acid. The organic phase was separated, washed with water, and brine (50ml of each), and the solvent evaporated in vacuo. The residue was dissolved in acetone (50ml), the solution stirred for 5 minutes with charcoal (ca. 1g). and the mixture filtered through a pad of kieselguhr. The filtrate was dried and evaporated to a foam (780mg), which was dissolved in ethyl acetate, and the solution run into petroleum ether to give the title acid as a colourless, amorphous solid (640mg, 63.3%); [.alpha.].sub.D.sup.23 - 64.degree. (c 1.01, acetone); .lambda..sub.max. (0.1 M-pH 6 phosphate buffer) 261 nm. (.epsilon. 19,200); .tau. (DMSO-d6) values include -0.11 (NH, d, J 8 Hz.), 4.00 (7-H, dd, J 5 and 8 Hz.), 4.70 (6-H, d, J 5 Hz), and 6.04 (CH.sub.3 --).
EXAMPLES 78-93
General Procedures for the Preparation of 7.beta.-(2-substituted oxyimino-2-arylacetamido)-3-(substituted)-ceph-3-em-4-carboxylic acids using Dicyclohexylcarbodiimide
(i) To a solution of a t-butyl (Example 93) or diphenylmethyl ester of 7.beta.-amino-3-(substituted)-ceph-3-em-4-carboxylic acid (1 equiv.) and dicyclohexylcarbodiimide (1-1.2 equiv.) in dry methylene chloride was added at 0.degree.-20.degree., a solution of the syn-2-alkoxy- or 2-aryloxyimino-2-arylacetic acid (1 equiv.) in dry methylene chloride. After stirring for 45 min.-3 hr. at room temperature the mixture was filtered and the filtrate was evaporated to dryness. The residue in ethyl acetate was washed successively with sodium bicarbonate solution, water and brine. The organic phase was dried and evaporated to a foam or solid. The resulting ester was purified by crystallisation or chromatography on silica and characterised by p.m.r. and thin layer chromatography.
Where the 7.beta.-amino starting material was available as its p-toluene sulphonic acid salt (Example 92) the free base was liberated by shaking with ethyl acetate and an excess of saturated sodium bicarbonate solution. After washing with water and brine the organic layer was evaporated to dryness and the free amine was redissolved in methylene chloride.
(ii) The intermediate esters so derived were deprotected by dissolving in a mixture of trifluoracetic acid (3-10 ml/1 g of ester) and anisole (1-5 ml/1 g of ester) and left at room temperature for 5-10 min. when the solvents were removed by evaporation under reduced pressure. The crude acid was shaken with ether or ethyl acetate and an excess of aqueous sodium bicarbonate and the aqueous layer was washed with ethyl acetate. The aqueous phase was separated, covering with ethyl acetate and acidified to pH 1-2 with 2N-hydrochloric acid. The organic layer was washed, dried and evaporated to give the required syn-7.beta.-(2-alkoxy-or 2-aryloxy-2-arylacetamido)-3-(substituted)-ceph-3-em-4-carboxylic acid, the compounds as obtained being listed in Table 6.
(iii) Diphenylmethyl 7.beta.-Amino-3-(1-ethyltetrazol-5-yl-thiomethyl)-ceph-3-em-4-carboxylate (used as Starting Material for Example 79)
(a) Diphenylmethyl 3-(1-ethyltetrazol-5-ylthiomethyl)-7.beta.-(2-thienylacetamido)-ceph-3-em- 4-carboxylate
A solution of diphenylmethyl 3-bromomethyl-7.beta.-(2-thienylacetamido)-ceph-3-em-4-carboxylate (11.67g) and 1-ethyltetrazoline-5-thione (2.6 g) in dry tetrahydrofuran (150 ml) was stirred at 0.degree. and a solution of triethylamine (2.8 ml) in tetrahydrofuran (25 ml) was added, dropwise, over 5 minutes. The resulting suspension was stirred for a further 25 minutes, during which time the temperature was allowed to rise to ca. 20.degree., and was then added, over 5 minutes, to a stirred mixture of 2N-hydrochloric acid (50 ml), water (350 ml), brine (350 ml), and ethyl acetate (600 ml). The organic phase was separated, and washed with 2 N-hydrochloric acid, saturated sodium bicarbonate, water, and brine, and dried and evaporated to a foam (11.71 g). A solution of this foam in benzene (25 ml) was purified by chromatography on kieselgel (200 g), with benzene:ethyl acetate= 5:1 as eluant. Appropriate fractions were combined and the solvents evaporated in vacuo. The residue was dissolved in methylene dichloride, and the solution run into petroleum ether to give the ester as a pale-yellow, amorphous solid (8.585 g, 68%); [ .alpha.].sub.D.sup.23 - 131.degree. (c 0.97, CHCl.sub.3); .lambda..sub.max. (EtOH) 264 nm (.epsilon. 7,300), .tau. (CDCl.sub.3) values include 4.12 (7-H, dd, J 5 and 9 Hz.), 5.04 (6-H, d, J 5 Hz.), 5.83 (CH.sub.3 CH.sub.2 --,q, J 7 Hz.), and 8.57 (CH.sub.3 CH.sub.2 --, t, J 7 Hz.).
(b) A solution of pyridine (2.13 ml) in dry methylene dichloride (10 ml) was added, over 5 minutes, to a stirred suspension of phosphorus pentachloride (5.47g) in methylene dichloride (20ml). After 10 minutes, the suspension was cooled to 0.degree., and a solution of diphenylmethyl 3-(1-ethyltetrazol-5-yl-thiomethyl)-7.beta.-(2-thienylacetamido)-ceph-3-em -4-carboxylate (8.3g) in methylene dichloride (40 ml) was added, dropwise, over 10 minutes. The resulting solution was stirred for a further 1 hour, during which time the temperature was allowed to rise to ca. 23.degree., and then added, over 10 minutes, to a stirred and cooled (0.degree.) mixture of methanol (30 ml) and methylene dichloride (60 ml). After 15 minutes, N-hydrochloric acid (50 ml) was added, and stirring continues for a further 10 minutes. The organic phase was separated, stirred for 30 minutes with saturated sodium bicarbonate (200 ml), and washed with saturated sodium bicarbonate and brine and treated with some charcoal. The suspension was filtered through a kieselguhr pad, the filtrate dried and evaporated to an oily solid (7.47 g). This material was triturated with ethyl acetate (15ml) to give the amine as a pale-yellow solid (3.854g, 58%); [.alpha.].sub.D.sup.23 - 213.degree. (c 1.14, CHCl.sub.3); .lambda..sub.max. (CHCl.sub.3) 265 nm. (.epsilon. 7,900); 268.5 nm. (.epsilon. 7,900), and 279.5 nm (.epsilon. 7,800); .tau. (DMSO-d6) values include 4.92 (d, J 5 Hz.) and 5.11 (d, J 5 Hz), (6-H and 7H), and 5.72 (q, J 7 Hz.) and 8.61 (t, J 7 Hz.), (CH.sub.3 CH.sub.2 --).
(iv) Diphenylmethyl 7.beta.-Amino-3(6-nitrobenzothiazol-2-ylthiomethyl)-ceph-3-em-4-carboxylat e, hydrochloride (used as Starting Material for Examples 81 and 85).
A solution of diphenylmethyl 3-bromomethyl-7.beta.-formamidoceph-3-em-4-carboxylate, 1.beta.-oxide (10.068 g) and 2-mercapto-6-nitrobenzothiazole (4.244 g) in dry N,N-dimethylformamide (125 ml) was stirred at 0.degree.; a solution of triethylamine (3.2 ml) in N,N-dimethylformamide (25 ml) was added, dropwise, over 5 minutes, and stirring continued for a further 2 hours, during which time the temperature was allowed to rise to ca. 23.degree.. The solvent was removed at 40.degree.(2mm.); the residue was partitioned between methylene dichloride (300 ml) and water (300 ml), and the organic phase washed with 2N-hydrochloric acid, water, and brine and dried and evaporated to a foam (14.35g). A solution of this foam in dry methylene dichloride (200 ml) was stirred at -20.degree., a solution of phosphorus tribromide (2.5 ml, ca 1.5 equiv.) in methylene dichloride (25 ml) was added, dropwise, over 5 minutes, and stirring continued for a further 30 minutes at -10.degree. to -15.degree.. The resulting solution was stirred for 30 minutes with saturated sodium bicarbonate (250 ml); the organic phase was separated, and washed with saturated sodium bicarbonate and brine and treated with some charcoal. The suspension was filtered through a kieselguhr pad, and the filtrate dried and evaporated to a foam (10.71 g). A suspension of this foam in methanol (200 ml) was stirred at 0.degree.; phosphorus oxychloride (3.66 ml - 2 equivs.) was added, over 2 minutes, and stirring continued for a further 2 hours at ca. 23.degree., during which time the solid all dissolved. The solvent was evaporated off; ethyl acetate (100 ml) was added, and the resulting suspension evaporated to dryness in vacuo. The residual gum was dissolved in methylene dichloride (100 ml); the solution was stirred at 0.degree. and ether (300 ml) was added, over 5 minutes. The precipitate was filtered off, and washed with ether (100 ml), and dried in vacuo to give the hydrochloride as pale-yellow flakes (9.744g, 78%); [.alpha.].sub.D.sup.23 - 94.degree. (c 1.0, DMSO); .lambda..sub.max. (EtOH) 328nm. (.epsilon. 13,400); .tau. (DMSO-d6) values include 4.72 (6H and 7-H), 5.10 and 5.57 (3-CH.sub.2, q, J 14 Hz.), and 6.10 (2 CH.sub.2).
(v) Diphenylmethyl 7.beta.-Amino-3-(6-nitrobenzothiazol-2-ylthiomethyl)-ceph-3-em-4-carboxyla te
A solution of the hydrochloride described above (910mg) in methylene dichloride (50ml) was stirred for 30 minutes with saturated sodium bicarbonate (50ml). The organic phase was separated, and washed with saturated sodium bicarbonate and brine and treated with some charcoal. The suspension was filtered through a kieselguhr pad, and the filtrate dried and evaporated to a foam (660 mg). This material was digested with methanol (50ml) to give the amine as a pale-yellow solid (432 mg, 50%); [.alpha.].sub.D.sup.20 - 195.degree. (c 1.01, CHCl.sub.3); .lambda..sub.max. (CHCl.sub.3) 251 nm. (.epsilon. 14,500) and 334 nm. (.epsilon. 16,800); .tau. (CDCl.sub.3) values include 5.04 and 5.22 (doublets, J 5 Hz., 6-H and 7-H), and 7.96 (NH.sub.2).
EXAMPLE 94
3-(5-Methyl-1,3,4-thiadiazol-2yl)thiomethyl-7.beta.-[2-(then-2-yl)oxyimino- 2-(thien-2-yl)-acetamido]-ceph-3-em-4-carboxylic acid (syn-isomer)
A solution of syn-then-2-yloxyiminothien-2-yl-acetic acid (715 mg) in methanol (;b 20 ml) was neutralised with 0.32 N sodium methoxide solution (8.7 ml). The solvent was removed in vacuo, and the residual sodium salt dried by azeotroping with benzene (2.times. 20 ml). Oxalyl chloride (685 mg) was added to a suspension of the above salt in benzene (30 ml) containing N,N-dimethylformamide (1 drop). The resulting solution was stirred at 20.degree. for 1.5 hr, and evaporated to dryness in vacuo to give the acid chloride.
A solution of this acid chloride in acetone (20 ml) was added dropwise to a stirred solution of 7.beta.-amino-3-(5-methyl-1,3,4-thiadiazol-2-yl)-thiomethylceph-3-em-4-car boxylic acid hydrochloride (1.00 g) in water (30 ml) containing sodium bicarbonate (;b 660 mg), at 0.degree.. The mixture was stirred for 2 hr. and allowed to reach room temperature. The acetone was removed in vacuo and the solution acidified to pH 1.9. The solution was extracted with ethyl acetate (6.times. 70 ml) and the combined extracts were washed with water, dried and evaporated to dryness. The crude product was washed with benzene cyclohexane (1:1, 2.times. 20 ml) and cyclohexane and dried to give the title carboxylic acid (863 mg) [.alpha.].sub.D - 61.degree. (c. 1.44, DMSO), .lambda..sub.max. (pH 6 buffer). 239 nm (.epsilon. 17,900), 273 nm (.epsilon. 19,150), .nu..sub.max. (Nujol) 1784 cm.sup.-1 (.beta.-lactam) .tau. (DMSO.sub.6) values include 0.09 (d,NH), 4.68 (s,CH.sub.2 -thienyl), 7.31 (s, CH.sub.3).
The sodium and potassium salts of the compounds of the Examples are prepared in conventional manner by reaction with sodium or potassium 2-ethylhexanoate respectively. To illustrate such salt preparation the formation of sodium 3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carb oxylate (syn-isomer is described in more detail below.
EXAMPLE 95
Syn-3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-c arboxylic acid (10.84 g.), was dissolved in the minimum amount of ethyl acetate (280 ml.) and a 1M solution of sodium ethyl hexanoate (25 ml.) was added dropwise to the stirred solution, which was stirred for 20 minutes, and cooled at 0.degree. for 2 hours. The precipitated salt was removed by filtration, and stirred with ethyl acetate (100 ml.) for 2 hours, and the solid (9.6 g.) filtered, washed and dried. This was then stirred with ether (2.times. 200 ml.) at room temperature for 30 minutes, producing the title compound (8.9 g., 78%), [.alpha.].sub.D.sup.22 + 60.degree. (c, 1.06 in water, .lambda..sub.max. (pH 6 buffer) 258-259 nm (.epsilon. 19,150).
The following example, illustrates the preparation of an amine base salt, such salts otherwise being prepared conveniently from the appropriate amine.
EXAMPLE 96
Diethanolammonium 3-Acetoxymethyl-7.beta.-(2-phenoxyimino-2-phenylacetamido)-ceph-3-em-4-car boxylate (syn-isomer)
To a stirred solution of crude syn-3-acetoxymethyl-7.beta.-(2-phenoxyimino-2-phenylacetamido)ceph-3-em-4- carboxylic acid (prepared as in Example 66) (5.94g) in 2-propanol (30 ml) was added a solution of diethanolamine (3.8 ml) in 2-propanol (8 ml). The precipitated solid was collected, washed (2-propanol, then ether), and dried in vacuo to give the title compound (3.43 g), m.p. 139.5.degree.; .tau. values (DMSO-d6) include -0.06 (d) (NH), 4.13 (dd) (7-H), 4.78 (d) (6-H), 6.96 and 7.96 (--CH.sub.2 --CH.sub.2 --).
The following Examples illustrate the formulation of pharmaceutical preparations.
EXAMPLE A
Dry Powder for Injection
Sterile sodium 3-acetoxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carb oxylate (syn-isomer) powder is filled into glass vials so that the glass vials contain 500 mg. and 1.0 g of the antibiotic as desired. Filling is carried out aseptically under a blanket of nitrogen. The vials are closed using rubber discs held in position by aluminum sealing rings, thereby preventing gaseous exchange or ingress of micro-organisms. The product is intended for reconstitution with water for injections or other suitable sterile vehicle shortly before administration.
EXAMPLE B
The last three ingredients are heated together at 150.degree. C. for one hour and then cooled to room temperature with stirring. The sterile antibiotic powder is added aseptically to this vehicle and the product refined with a high speed stirrer. The preparation is filled aseptically into sterile collapsible aluminum tubes so that each tube contains 3.0 g of the preparation comprising 300 mg. of the cephalosporin derivative.
EXAMPLE C
The product is intended to reconstitution with sufficient purified water to give a final volume of 100 ml., which should all be administered within a few days, each 5 ml. dose of syrup containing 250 mg. of the cephalosporin derivative.
In order to prepare the blend, the amaranth is intimately mixed with some of the sodium citrate and milled. The sodium saccharin and citric acid are blended together. They are mixed thoroughly with the color triturate, then with the remainder of the sodium citrate, flavor and antibiotic powder in that order. This blend is milled, mixed with sucrose and then 75 g. is filled into each of a number of 150 ml. capacity bottles, which are closed with moisture-proof screw caps.
EXAMPLE D
(a) Oral Capsules
Sodium 3-acetoxymethyl-7.beta.-(2-ethoxyimino-2-phenylacetamido)ceph-3-em-4-carbo xylate (syn-isomer) is blended with one percent magnesium stearate and filled into size 0 hard gelatin capsules, each capsule containing a claimed dose of 240 mg. of the cephalosporin antibiotic. The capsules are packed in glass vials with plastic caps giving a moisture-proof seal.
Claims
We claim:
1. A compound selected from the group consisting of a cephalosporin antibiotic highly stable to .beta.-lactamases having the formula ##STR148## wherein R is phenyl, thienyl or furyl and R.sup.a is C.sub.1 -C.sub.8 alkyl or benzyl; and a physiologically acceptable salt thereof.
2. The compound of claim 1 which is sodium 3-hydroxymethyl-7.beta.-(2-methoxyimino-2-phenylacetamido)ceph-3-em-4-carb oxylate (syn isomer).
3. 3-Hydroxymethyl-7. beta.-(2-benzyloxyiminothien-2'-ylacetamido)ceph-3-em-4-carboxylic acid (syn isomer).
Patent Citations (2)
| Patent | Date | Inventor | Cited By |
|---|---|---|---|
| DT2,204,060 | 1972-08-01 | ||
| DT2,223,375 | 1972-11-01 |
Non-Patent Literature (2)
- Gregson et al., Chem. Abst., vol. 77; 126, 662a, (Abst. of L).
- Cook et al., Ibid. vol. 78; 58,444z, (Abst. of M).