PHYSIOLOGICALLY ACTIVECO~MPOUSDS. V
May, 1963
231
TABLEI1 AMIXOTHIOL ESTERHYDROCHLORIDES OF HYDROXY ACIDS,RR’C(OH)COS( CH2),NR2” HC1 Compound
R”
R’
R
Yield,
M.U.,
%
OC.
z
3101. formula
-Analyses, ?;.--------.--Calculateii-----Found--C H C
61.2% 6.30 ClsHy&lXOyS 61.43 153-154 2 7 C6H5 C& CHI 83.31 6.00 C ~ O H ~ ~ C I S O 63.2% ~S 138-139’ 23 111 C6H5 C6H5 C2H5 2 b 64.06 2 17 185-187 6.69 C ~ I H & ~ ~ O , S64.35 112 CsHj C& 113 CsH5 CsH5 CHa 3 1gC 180-182 CigHuClX0,S 62.36 6.61 62.3% 114 C6135 3-CHzCsH4 CyH5 2 34 140-141 CPIH&lSO?S 64.02 7.16 63.68 115 CsHj 3-CHgCsH4 CH3 :3 24 145-147 C9oHyaClSOrS 63.2% 6 . 9 0 63.24 CsHj 3>4(CII3),C& C& 2 10 155-157 CyrHz,CISOyS 64.76 7.41 64,G-l 116 117 CsH5 3,4-CH202CsH, CyHj 2 23 154-156 C,1Hy&lSO$ 59.49 6.18 59.30 118 CsHj :3,4-CHyOnCsH3 CHI 3 11 179-181 CyLH&1SO4S 58.59 5.90 5s.7 2 11‘3 4-CH3CsH4 4-CH,C6Ha CyHj 2 6jd 166-16i CryH30ClS02F 64.76 7.41 64.91 120 CsHj CjH, CyHj 2 3 j d 150-151.5 Ci!iH,oCl?iO?S 61.35 8.13 61.14 121 CsHj CsHii CrHj 2 86d 174-175.5 C?,H3?C1S0.S 62.23 8.36 62.% Zhur. Obshchei. Khin~., 28,635 (1958). This compound containing 835is reported, m.p. 143-146”, C. A , , 52, 171.55 (1958); dine ester. Free base. d Based on corresponding a-chloro derivative. 110
Xa
RCOR’ +R(R’)C(ONa)Sa
CO [SCH2CH2N(C%Ho)21z f
liquid S H 3
llquid S H 3
SHaCl
R(R‘)C(OSa)C(OXa)[ S C I I y C H ~ SClH,)?]r ( -+ llquld NIII
R( R’)C(OH)COSCHyCH,N( CyHj)2
2-S,S-Diethylaniinoethyl thiol benzilate was produced in 627, yield by this method. Unfortunately, slight variations in the nature of R and R’ led to unsatisfactory results. 2,3-Din~etliylbenzoplieiioIie, for example, gave a product which had the composition of the corresponding carbinol. 4,4’-Dimethylbenzophenone gave a product which corresponded to I on the basis of its analysis and infrared spectrum. hcetophenoiie as well did not give the desired ester, [(4-CH3C6H4),C(OH)IsCO, perhaps because of the formation of the sodium salt of the enolic form. 3IYDRI.ITIC ACTIVITY NO.
102 103 105 106 108 109 110 111 113 114 115 116 117 118 119 120 121
Vjdilatir actirity“
100
1/60 1/30 1/25 1/60 1/60 1/100 1/60 1/60 1/60 < 1/250 1/ J 1/ 2 1/25
0 0 0 0 -I+ 4+ 4+ 4+ 4+
2+ 4+ 4+
3+ I+ 4+
4+ 4+
50
25
11 5
6.M 7.56 6.40
5 .S1 7.4%
Piperi-
starting with the appropriate substituted acetic acid and proceeding as indicated previously for compound 109 in Table I. The salts in Table I1 were obtained from the corresponding chloro derivative essentially by the met hod of Kolloff, et C X ~ . followed ,~ by hydrolysis. The free base of the 0-hydroxythiol esters, after purification, was converted into the hydrochloride. Pharmacological Results and Discussion.-The mydriatic activity and cerebral stimulation tests, as listed in Table 111, were made in the laboratories of Parke, Davis and Company through the courtesy of Dr. Nartin L. Black. The mydriatic assay method involves estimation of the effect of the drug on mouse pupillary size and the duration of this effect relative t o the effect and duration produced by atropine sulfate, both being given intramuscularly. For the C S S tests the “jiggle cage”
TABLEI11 CEREBR4L S T I \ I L L L T I O \
------------CN~
0
< 1/a50
4\D
H
6.45 6.90 6.47 6,s;) 7.19
OF
THIO]. .\CID ESTERS
a c t n i t % a t indicated dosage (nig /Ag-------)----b 6 3 15 075 038
030
010
005
I+
0
0 0 0 0 I+ 4+
2+
I+
a+
4+ 4+
4+
4+ 4+ 4+ 3+ 3+ 4+ 2+
4+ 4+ 3+ 2+ 4+ 2+
4+ 4s
4+ 3+ 2+
4+ 4+ 4+ -I+
4+ 4+ 4+ 4+
4+
4+
4+
4+
-I+ 4+ 1+
-I+
4+ 1+
4+ 0
2+
0
4+ 4+
4+ 4+
4+ 1+
3+
1+
%+
0
0 0
4+
4+
2+
0
4+
4+ 4+ 4+
I+ 4+ 4+ 4+
4+ 4+ 4+
4+ 4+
4+ 4+ I+4+ 4+ 4+
’
Effectiveness in extent and duration of dilation of the pupil compared with atropine sulfate as 1 (mice). Three rats (positive controls) were each given a subcutaneous injection of 30 mg./kg. of caffeine and each of 3 negative control animals received the same volume of isotonic saline solution subcutaneously. One of the doses of the test drug was then given to each of 3 test animals; subcutaneously if soluble, orally if not. Recorder responses were counted for 30 min. intervals for the ensuing 2 hr. period and the one that represented peak activity was chosen for analysis. If one lets S,, N,, and S , represent, respectively, the number of responses for treated, positive control, and negative control animals, a score of 1+ to 4+ can he assigned: ( S , - AVr)/(AYp- S , ) X 100 = % R. Benactyxine. The score corresponding to % R ranges is 76-100, 4+; 51-75, 3 + ; 30-50, 2 + ; and 20-30, l + . a
”as achieved in preparing the chloroacid chlorides of acids containing a cycloalkyl group by
(6) H. G. Kolloff,.J. H. Hunter, E. H. Woodruff, and R . B. Moffett, J . A m .
men.sot., 71, 3988
(1949).
May, 1963
DERIVATIVES OF ( -)-trans-2,3-E~oxrarrcc1~1~ ACID
a-Chloroacid Chlorides of Substituted Benzilic Acids.-3hIethylpheny1 phenyl-, 3,4-dimethylphenyl phenyl- and 3,4methylenedioxyphenyl phenylchloroacetyl chlorides3: I n these cases, distillation under reduced pressure led t o decomposition of the final products. They were therefore freed from ether, subjected t o high vacuum for 15 min., and employed xvithout further purification. Phenylcyclopentylchloroacetyl Chl~ride.~-Phenylcyclopentyl:lcaetir acid (10 g.) gave 9.8 g. (787,) of chloroacid chloride, b.p. 124" (0.5 mm.) f n a l . Calcd. for C13H14C120:CI, 27.68. Found: C1, 27.80. Ester Hydrochlorides of Substituted Hydroxyacids in Table 11. 3-N,N-Dimethylaminopropylisothiouroniunichloride hydrochloride s a s obtained in 83' yield from 3-chloro-5,X-dimethylpropylamine hydrochloride by the method of Albertson and Clinton,*1 m.p. 159-161'. dnaI. Calcd. for CcHliCl?N3S: C, 30.77; H, 7.32. Found: C, 30.94, H, 7.16. 2-N,N-Dimethylaminopropanethio121 was used in the ethereal solution obtained on extraction. Compounds 110 through 118 in Table I1 were prepared by identical procedures viliich may be described by the synthesis of (21) N. F. Albertson and R . 0 . Clinton, J . A m . Chem. S o e . , 6 7 , 1222 (194.5).
233
3-N,N-dimethylaminopropylthiolbenzilatehydrochloride according to the method of Kolloff, et al.8 T o a solution of 8 g of diphenylchloroacetyl chlorided in 50 ml. of anhydrous ether, \vas added the ethereal extract from the alkaline hydrolysis of 7 g. of 3-S,Sdimethylaminopropyl isothiouronium chloride hydrochloride; an oil precipitated immediately. After refluxing for 1 lir , the reaction mixture was cooled and the ether was decanted The residual oil was heated on the steam bath for 15 min. with 100 nil. of n ater and one drop of concd. hydrochloric acid. t-pon being cooled and made basic with sodium carbonate, a white solid separated. Several crystallizations from ethanol led to m.p 86-87". When hydrogen chloride was passed into A solution of the solid in anhydrous ether, 1.8 g. ( 1BCb)of a white solid formed, 11hich melted at 180-182" after several crystnllization from ethanol-ether . Compounds 119, 120 and 121 were prepared from the corresponding a-cliloro derivatives by a procedure \\ liicli may be illustrated for 2-N,N-diethylaminoethyl phenylcyclohexylthiolglycolate hydrochloride. The a-chlorester hydrochloride ( 2 3 g.), dissolved in a minimum amount of nater, was refluxed for 2 Iir , made dlstinctly alkaline M ith sodium carbonate solution, itnd extracted with ether. The dried ethereal solution was acidified with ethereal hydrogen chloride to give a solid uhicli, \\hen crystallized from a mixture of acetone, methanol, and ether, weighed 2.0.5 g. ( 8 6 5 ) m p. 174-11.5 5".
Derivatives of ( -)-trans-2,3-Epoxysuccinic -4cid and Some of their Biological Effects l l a x W. MILLER .l/erlical Research Labomtories, Charles Pfiterand Po., I n c . , Groton, C'onn.
Received October 29, 1962
Syninietrical esters and amides as well as the nitrile have been prepared from the mold metabolite, ( - ) - t r a m ?,&epoxysuccinic acid. Opening of the oxirane ring in epoxj-succinic acid and its methyl ester with aninionia and amines t o form e,.ythro-p-hydroxy-L-aspartic acid and S-substituted analogs is discussed and some products are described.
The three isomers of epoxysuccinic acid (transracemate and cis-meso) have been related to the corresponding tartaric and chloronialic acids. The cis-meso form has been prepared by hydrogen peroxide oxidation of benzoquinone4 and by nitric acid oxidation of the macrolide antibiotic, carbomycin,j and both the cis isomer and the trans racemate can be prepared by tungstate-catalyzed hydrogen peroxide oxidation of, respect,ively, maleic and fumaric acids.6 To obtain the pure (-)-trans isomer, however, a fermentat'ive preparation was used since fairly high yields had been reported from t k 2 fermentation of glucose with d s p e r yillus jumigat~s.~-'O We observed yields from this mold of over 20 g. '1. of fermentation broth or a 40% molar conversion calculated from the glucose moiety of' ( 1 ) R. Kulin and F. Ehel, Ber., 68, 926 (1925). (2) R . Kuhn and R . Zell, ibid., 59, 2514 (1926). ( 3 ) R . Iiuhn and T. Wagner-Jauregg, i b i d . , 61, 513 (192b). (1) E. Weitz, H.Schohbert, and H. Seibert, ibid., 68, 1166 (1935). ( 5 ) R. B. Woodward, Angezo. Chem., 69, 50 (1957). (6) G.B. Payne and P. H. Williams, J . 0 ~ g C. h e m . , 24, 54 (1959). ( 7 ) J. H. Birkinshaw, -4.Bracken, and H. Raistrick, Biochem. J . , 39,7 0 (1945). (8) J. hloyer, U. S. Patent 2,674.561, Sept. 8, 1950 (to Secretary of Department of Agriculture). (9) IT. Martin and J. Foster, J . Bacteriol., 70, 405 (1955). (10) It is interesting t h a t fumagillin another Aspergillus fumigatus metabolite, contains two epoxide groups: [J. Landquist, J . Chem. Soc., 4237 (19561, and D. 9. Tarbell, R . 11. Carman, D. D. Chapman, K. R. Huffman, and N. .J. l\lcCorkindale, J . Am. Chem. Soc., 8 2 , 1005 (1960)l.
the sugar in the crude molasses used. Other inolds, particularly Penicillium itiniferum and Jlorzilia jmmosa also produce this acid."-'j The oxiraiie ring as substituted in epoxysuccinic acid is less reactke to acidic reagents than many epoxides, l6 permitting selective reactioiis a t the carboxyl groups. Thus, either free epoxysucciiiic acid or its slightly soluble bariuni salt, the form in vhich the acid was isolated from fermentation broths, could he esterified in alcohols with sulfuric acid catalyst. .I number of the esters so prepared are listed in Table I. K h e n (-)-trans-epoxysuccinic acid \vas heated \\-ith hydrochloric acid in methanol, dimethyl er.~jtlz~-o-chloromalate was formed. Treatment of either this ester or dimethyl epoxysuccinate with cold aqueous ammonia led to high yields of the slightly soluble (-)-transepoxysuccinainide. Similarly other amides were pre(11) K. Sakaguohi, T. Inoue, and Y.Tada, J . A g r . Chem. S o c . J a p a n , 13, 241 (1937); Proc. I m p . Acad. (Tokyo), 13, 9 (1947). (12) Ii. Sakaguchi, T. Inoue, and Y.Tada, J . .4gr. Chem. SOC.J a p a n , 14, 362 (1938). (13) K.Sakaguchi and T. Inoue, i b i d . , 14, 1517 (1938). (14) K. Sakaguchi, T. Inoue, and S. Tada, Zentr. Bakf. Parasiferik., I1 .Ibt., 100, 302 (1939). (15) I(. Sakaguchi and T. Inoue, J . 4 g i . Chem. SOC. J a p a n 16, 1013 (1940). (16) For quantitatire determinations of the rates of cleavage of a number
of epoxysuccinic acid derivatives in pyridinium hydrobromide -acetic acid a t 30° see hl. W.Miller, J . O r g . Chem., 2 8 , 1148 (1963).
