The Synthesis and Oxidative Rearrangement of ... - ACS Publications

carbornetl1oxy-2,2-dimethyl-5-oxo-6~-phenyl~cetamidoperhydro-l,4-thiazepine (SIII) while .... to yield (72%) 3~-carbomethoxy-2,2-dimethy1-5-0~0- .... ...
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Dec. 5 , 1964

1, ~ - T H I A Z E P I XRELATED ES

pounds have been reported with varying results. 2 , 2 0 In the iodination of N-acTY and N-acMIT, significant salt effects are not observed a t pH 7 11 and are small a t pH T 90 and 9 45. These results and the variance of the previous reports suggest that the salt effects are secondary The fact that the effects on N-acTY and Y - a c l l I T are of a similar magnitude further suggests that the equilibria displaced by the salts may well be on the buffer constituents rather than the reactants

[CONTRIBL-TIOS FROM

THE

TO PENICILLIXS

5307

themselves According to Debye-Huckel theory for dilute soutions, the slope of a plot of log k against pl" depends in part on the product of the ionic charge of the reactants. This seems quite unlikely for more concentrated solutions Acknowledgment.-\Ye gratefully acknowledge the help of Dr. Nones Rerman in deriving eq. 12, in establishing our computer procedures, and for helpful criticism,

SOYES CHEMICAL LABORATORY, ~-SIVERSITY

OF

ILLISOIS, URBAYA,ILL ]

The Synthesis and Oxidative Rearrangement of Some 1,.l-Thiazepines Related to the penicillin^'-^ BY NELSONJ. LEONARD A N D G . EDWIAT~ILSOS,

JR.~

RECEIVED ACGUST3, 1964 LVe have studied the synthesis, stereochemistry, and oxidative rearrangement of some 1,4-tliiazepines related (XIII) t o the penicillins 3~-Carbomethoxy-2,2-dimethyl-5-oxo-6~-phenylacetan1idoperhy~ro-l,4-thiazepine was formed by the reaction of a-phenylacetamidoacrylic acid and D-penicillamine methyl ester hydrochloride in acetonitrile in the presence of dicyclohexylcarbodiimide and triethylamine, b u t none of the 3 ~ - 6 ~ - d i a s t e r e o m e r was isolated. Proof of the induced configuration 6n was provided by desulfurization of XI11 with Raney nickel to give methyl N-( S'-phenylacetyl-D-alanyl)-D-valinate.The stereoisomer synthesized ( X I I I ) is the thermodynamically more stable form. The oxidation of XI11 by nieans of chlorine produced 3n-carbomethoxy-~,2-dimeth~~l-~-oxo-6-phenylacetamido-2,3,4,5-tetrahydro-l,4-thiazepi1ie (Xl-1) and two isomeric 3-isothi( X S I ) and methyl a-isopropylazolones, methyl a-~-~sopropenyl-4-phenylaceta1n~do-3-~soth~azo~one-~-acetate ide11e-l-phenylacetamido-3-isothiazolone-2-acetate ( X X I I ) . The oxidative formation of 3-isothiazolones was shown t o have some generality. Photoreduction of XVI in ethyl mercaptan solution a t 55' regenerated 3 ~ carbornetl1oxy-2,2-dimethyl-5-oxo-6~-phenyl~cetamidoperhydro-l,4-thiazepine( S I I I ) while photoadducts of ST-I were formed a t 20" with ethanol, 2-propanol, and ethyl mercaptan (S\-11-SX).

II ile an advanced stage has been reached in the elucidation of the intermediates involved in the pathway of penicillin b i o ~ y n t h e s i s , ~including the isolation of the tripeptide y-( a-aminoadipyl)cyst(e)inylvaline from the mycelium of Penacdlaum chrysogenum, the place in the sequence where oxidative condensation between the @-positionof the cyst(e)ine moiety and the peptide nitrogen atom occurs has not been determined Lye considered that the earlier suggestions of the transannular formation of the C-K bond aeross a substituted 1,4-thiazepine derivative (I + 11)7-9 had n o t as yet H RCOHNWS~~ O;VI) with t h a t of an authentic sample and the optical rotation, in 3 ml. of chloroform was added 134 mg. (1 mmole) of N-chloro[ a I z 5-39.3' ~ (0.57, in E t O H ) , was compatible with the value succinimide and a trace of dibenzoyl peroxide. T h e mixture was of -39.7" obtained previously. The melting point of this swirled until a slight temperature increase was noted, then it was material, 180-182', was somewhat higher than the original, left to stand for 2 h r . Analysis of the crude reaction mixture by suggesting t h a t the further reaction had discriminated against a n.m.r. showed t h a t the main product was methyl a-D-isopropenyltrace of impurity consistently present in XI11 obtained from peni4-phenylacetamido-3-isothiazolone-2-acetate ( X X I ) with smaller cillamine methyl ester hydrochloride. amounts of minor side products and iinreacted starting material. Pyrolysis of the Ethyl Mercatpan Adduct of 3n-CarbomethoxyT h e chloroform solution was washed with water, dried over 2,2-dimethyl-5-oxo - 6 -phenylacetamido-2,3,4,5tetrahydro- 1,4sodium sulfate, and evaporated t o dryness under vacuum. T h e thiazepine, M.p. 205.5-206So.-The ethyl mercaptan adduct, semicrystalline solid remaining was dissolved in 5 ml. of absolute m .p. 205.?~-206.5~, of 3~-carbomethoxy-2,2-d~methyl-5-oxo-6- ethanol and allowed t o crystallize a t -20", giving 168 mg. (49 70) phenylacetamido-2,3,4,5-tetrahydro-l,4-thiazepine( X V I I ) , 80 of fine, slightly tan needles of X X I , m.p. 146.5-151'. mg., was maintained at a temperature of 210" under a current of Isomerization of Methyl a-~-Isopropenyl-4-phenylacetamido-3nitrogen for 1 hr. T h e remaining brown melt, cooled and disisothiazolone-2-acetate (XXI) to Methyl a-Isopropylidene-4solved in deuteriochloroform, showed the n.m.r. spectrum of 3D50-mg . carbomethoxy-2,2-dimethyl-5-oxo-6-phenylacetamido-2,3,4,5-te- phenylacetamido-3-isothiazolone-2-acetate (XXII).-A sample of methyl a-~-isopropenyl-4-phenylacetamido-3-isothiazotrahydro-1,4-thiazepine ( X V I ) , indicating t h a t the total amount lone-2-acetate ( X X I )was dissolved in 0.8ml. of chloroform t o which of impurities was no more than 5%. Chromatographic purificawas added 0.01 ml. of triethylamine and 3 drops of water. T h e tion of this material on 2 g. of silica gel with an ether eluent afmixture was swirled, then allowed t o stand a t room temperature forded, after crystallization from ethanol, 20 mg. (307,) of slightly for 2 hr. T h e volatile fractions were removed; a n n . m . r . spectan crystals of X V I , m.p. 140-143'. trum in deuteriochloroforrn showed t h a t complete isomerization Photoreduction of the Ethyl Mercaptan Adduct of 3n-Carbot o methyl a-isopropylidene-4-phenylacetamido-3-isothiazolone-2methoxy-2,2-dimethyl-5-oxo-6-phenylacetamido-2,3,4,5-tetrahyacetate ( X X I I ) had taken place. After removal of the deuteriodro-l,4-thiazepine (XVII), M.p. 205.5-206S0, in Ethyl Mercapchloroform, crystallization of the crude material from ethanol tan --A suspension of 48 mg. of the ethyl mercaptan adduct of 3~-carbon1ethoxy-2,2-dimethyl-5-oxo-6-phenylacetamido-2,3,4,5-provided 30 mg. (60%)of colorless needles of the isomerized prodtetrahydro-1,4-thiazepine( X V I I ) , m.p. 205.5-206.5', in 3 ml. of (40) Alternatively the reaction mixture could he allowed t o stand for 24 ethyl mercaptan was irradiated by a Speri sun lamp, Model Phrs. without ultraviolet iri-adiation, b u t with 2 0 mg. of dibenzoyl peroxide 109, a t a distance of 2 in. from the arc for 48 hr. The solvent was added. These conditions have been reported t o give a compound erremoved, and the product, dissolved in deuteriochloroform, was roneously assigned structure XVl.14 analyzed by n . m . r . T h e n.m.r. spectrum showed the crude (5.0)Spontaneous crystallization will t a k e place after about 1 week a t material t o be mainly 3~-carbomethoxy-2,2-d~methyl-5-oxo-6~- - 2 0 0 .

5316

NELSONJ. LEONARD AND G. EDWIN WILSOK,JR.

uct, m.p. 232-233". The m.p. and mixture m.p. were identical with t h a t of a sample of X X I I obtained a s described below. Methyl ~-Isopropylidene~-phenylacetamido-3-isothiazolone-2acetate (XXII).-To a solution of 4.0 g. (0.01 mole) of 3 ~ - c a r b o methoxy-2,2-d~methyl-5-oxo-6~-phenylacetamidoperhydro1,4thiazepine ( X I I I ) and 2.0 g. (0.015 mole) of N-chlorosuccinimide in 30 ml. of chloroform was added with swirling 20 mg. of dibenzoyl peroxide. After a n induction period of about 10 min. a sharp rise in temperature was noted. After 3 h r . the yellow solution had returned t o room temperature and no starting material was detectable by thin layer chromatography on silica gel. The chloroform solution was washed with 30 ml. of water to remove succinimide, dried over sodium sulfate, and applied, in a total volume of 46 ml. of chloroform solution, to a column made from 400.g. of F l o r i d , 5.5 X 30 cm. Elution was carried out with ether-ethyl acetate mixtures and the optical density a t 295 mp was recorded for each 70-ml. fraction. Fractions 1-14 contained no product. From fractions 15-35 were isolated 0.82 g. (21yc) of crude methyl a-isopropylidene-4-phenylacetamido-3isothiazolone-2-acetate ( X X I I ) as colorless needles. Recrystallization of this material from ethanol, after washing a chloroform solution of this material with water to remove any succinimide, gave an analytically pure sample, m .p. 232.2-233.1 a ; Y::: 3270, 1710, 1680, 1619, 1572, 1525, and 1489 c m . 7 ; 295.5 mp ( e 10,020) with a shoulder a t 307 mp ( e 7900); [ a I z 5d~2 ' ; n.m.r. 7-values: 1.02, 1.30, 2.68, 6.24, 6.37, 7.82, and 8.19 p.p.m. Anal. Calcd. for CliHtsYsOaS: C, 58.94; H , 5.24; S , 8.09; mol. wt., 346.40. Found: C, 58.90; H, 5.31; X, 8.23; mol. wt., 346 (mass spectrum, molecular ion). Fractions 40-65 afforded 1.6 g. (40yc) of 3~-carbomethoxy-2,2-dimethyl-5-oxo-6phenylacetamido-2,3,4,5-tetrahydro-l,4-thiazepine ( X V I ) as an impure oil which could be crystallized from ethyl acetate-hexane. cis-(3-t-Butylthio)acrylamide (XXIV).-To a solution of 4.0 g. (0.058 mole) of propiolamide and 5.2 g. (0.058 mole) of t-butyl mercaptan in 20 ml. of ethanol in a flask surmounted by a condenser was added 5 mg. of sodium methoxide. The system was closed from the atmosphere until the initially exothermic reaction had subsided, approximately 2 min. Removal of the solvent after cooling the reaction left a soft, orange solid. Crystallization of this mass from methylene chloride-hexane a t Dry Ice temperature gave 5.8 g. of slightly yellowish prates of cis-(3-tbutylthio)acrylamide, m.p. 159-160.5'. Concentration of the mother liquors afforded a n additional 1.2 g. of the cis isomer, raising the yield t o 76%; Y~~~~ 3050, 3465, 3390, 3315, 3150, 1664 (amide C=O), 1590, and 1570 cm.-l (amide C=O); X2p 215 (e2060)and282mp(c13,000); n.m.r.s-values(CDCI8):2.93 d ( J = 10.5c.p.s.),4.13d, a n d 8 . 6 2 p . p . m . .4nal. Calcd. for C7H13NOS: C , 52.79; H , 8.23; X , 8.80. Found: C , 52.87; H, 8.23; N,8.86. T h e mother liquors from the second crystallization were evaporated t o dryness. Hexane extraction of the remaining solid gave some material identified by n.m.r. as the trans isomer, doublets a t T 2.20 and 4.03 p.p.m. ( J = 15.0 c.P.s.), and a singlet a t 7 8.58 p.p.m. This material crystallized poorly from hexane, m . p . 76-76.5", and it decomposed upon attempted sublimation. 3-Isothiazolone.-To a solution of 2.5 g. (15.7 mmoles) of cis(3-t-buty1thio)acrylamide ( S X I Y ) in a mixture of 40 ml. of chloroform and 10 ml. of ethyl acetate a t -60" was added dropwise and with stirring 31.5 ml. (15.7 mmoles) of chlorine, 0.5 114 in carbon tetrachloride. After 1 hr. a t this temperature, the mixture was warmed to reflux while a current of nitrogen was bubbled through it to remove the hydrogen chloride formed. After 2 hr. the elimination of hydrogen chloride had ceased. The solvent was replaced by 30 ml. of water, and 10 ml. of 2 Nsodium hydroxide solution was added. The aqueous solution was ex-

Vol. 86

tracted twice with ether, carefully neutralized, and extracted continuously with ether for 2 h r . T h e ether solution was dried over sodium sulfate and evaporated in vacuo to leave 0.38 g . (24Yc) of a yellow oil. T h e oil was partly dissolved in 20 ml. of water, filtered, and the aqueous solution was extracted continuously with ether for 2 hr. Removal of the ether solvent after drying left crystalline 3-isothiazolone. Recrystallization from hexane and two sublimations afforded an analytical sample, m.p. 73-74'; Y:G( 3090, 3030, 2980, 2800, 2690, 2630, 2540, 1659, 1639, 1573, 1546, 1420, 1328, 1072, 988, 868, 832, 681, and 602 cm.-'; "$A: 257 mp ( e 6400) with a shoulder at 280 mp ( e 1350); in base a shift to 279 mp ( e 6750) was observed for the maximum. 3-Isothiazolone showedn .m.r. signals a t r-values (CDC13) -1.90, 1.56 d ( J = 5.0 c.P.s.), and 3.42 d p.p.m. Anal. Calcd. for C3HaSOS: C, 35.62; H , 2.99; Pi, 13.85. Found: C , 35.55; H , 3 . 0 0 ; X, 13.78. General Procedure for Degradation of the 1,4-Thiazepines and 3-Isothiazolones to Amino Acids .-A 10-mg. sample of material was heated under reflux with ca. 1 g. of Raney nickel W-2 in about 10 ml. of absolute ethanol for 2 hr. T h e Raney nickel was washed three times with absolute ethanol by decantation before use. A check was made before and after the reaction to make sure the catalyst was pyrophoric. T o the residue remaining after gravity filtration of the reaction mixture and evaporation of the ethanol under vacuum on a rotary evaporator was added 1 ml. of 8 N sulfuric acid, and the solution was heated under reflux for 4 hr. Xeutralization of the cooled reaction mixture to p H 6 with barium hydroxide, vacuum filtration, and lavation of the precipitate with hot water was followed by removal of the water under vacuum on a rotary evaporator t o a volume of ca. 1 ml. This solution was then subjected t o thin layer c h r ~ m a t o g r a p h y on ~l silica gel G with 1-butanol-acetic acid-water (60: 15:25) and phenol-water (75:25), on cellulose M N 300 G (Merck) with phenol-water (75 :25), and to paper chromatography with phenolwater (80 : 20) and 1-butanol-acetic acid-water (60 : 15 : 2 5 ) . Valine and alanine were chromatographed together with the unknowns as standards. Double spotting was carried out t o make sure t h a t the Rr values of standard and unknowns did not change each other. Valine could be distinguished from a-aminobutyric acid by t.1.c. on silica gel with phenol-water (75:25). Thin Layer Chromatography.-Thin layer chromatograms were run on silica gel G plates (Merck) equilibrated with atmospheric moisture and developed with ether eluent. They were then sprayed sequentially with three developing reagents in the order given: reagent 1, 0.4% disodium fluorescein in water; dark spots on a fluorescent yellow background were seen under a long wave length ultraviolet lamp. Reagent 2, 17' potassium permanganate in 5Yc sodium carbonate solution; yellow spots on a purple background were observed. Reagent 3, Dragendorff reagent; brown spots on a yellow background were observed. This reagent was prepared by dissolving 7.1 g. of bismuth subcarbonate in 20-25 ml. of 30Yc nitric acid and adding this solution slowly and with stirring to a hot solution of 28 g. of potassium iodide and 1 ml. of 6 N hydrochloric acid in ca. 5 ml. of water. The stock solution is prepared by cooling the above t o about 1 5 O , filtering, and diluting to 100 ml. The spray solution is made by adding in order 5 ml. of 6 N hydrochloric acid, 20 ml. of water, and 2 ml. of stock solution.52 ( 5 1 ) K. Randerath, "Thin Layer Chromatography," Academic Press, Inc., New York, N . Y., 1963, p , 93 IT. ( 5 2 ) This procedure, obtained from Dr. Tozo Fuji, is a modification of t h e one found in R . J. Block, E. L. D u r r u m , and G. Zweig, "A Manual of Paper Chromatography and Paper Electrophoresis," Academic Press, I n c . , Xew York, N . Y., 1958, p. 360 f .