Sept., 1939
COMMUNICATIONS TO THE EDITOR
2563
A detailed report of our work will be presented in the future. We acknowledge gratefully valuable suggestions of Dr. H. Gilman and Dr. F. B. LaForge. We are also indebted to Merck & Co., Inc., for generously furnishing us with a supply of the pyrimidine derivative used in this work. The analytical work was carried out by Dr. Carl Tiedcke of New York City.
The potency of Z-methyl-l,4-naphthoquinone (Thayer-Doisy units) agrees with the value previously assigned to the natural K1, namely, 1000 units per milligram [Proc. SOC.Exp. Biol. Med., 41, 194 (1939)l. These results also confirm the findings of Ansbacher and Fernholz [THIS JOURNAL,61, 1932 (1939)l. Incidentally, in view of these observations and the lack a t this time of a suitable standard, it is that 2-methyl-l,4-naphthoquinone RESEARCH LABORATORY FRANZ C. SCHMELKESsuggested WALLACE & TIERNAN PRODUCTS, INC. ROBERT R. JOINER should be adopted as a basic standard for the BELLEVILLE, NEWJERSEY assay of vitamin K. This compound has the RECEIVED JULY 3, 1939 desirable qualities of a standard in that it can be obtained readily in a satisfactory state of purity, has a definite melting point for characterization, VITAMIN K POTENCIES OF SYNTHETIC COMPOUNDS and when protected from excessive exposure to Sir: light is relatively stable. The unit could then be defined in the terms used by the League of NaI n view of the failure of the absorption of many tions committee as the specific vitamin K activity patients in which vitamin K therapy is highly desirable, we have been examining various com- of one microgram of pure 2-methyl-l,4-naphthopounds which could be administered intravenously quinone. in aqueous solution. The most active com- BIOCHEMISTRY DEPARTMENT S.A . THAYER SCHOOL OF MEDICINE S. B. BINKLEY pound found is 1,4-dihydroxy-2-methylnaphtha-SAINT LOUISUNIVERSITY D. W. MACCORQUODALE lene which has a potency of approximately 1000 SAINTLOUIS,MISSOURI E. A. D o r s ~ OF A. D. EMMETT RESEARCH LABORATORIES Thayer-Doisy units per milligram. It can be PARKE, DAVISAND COMPANY RAYMOND A. BROWN prepared readily by the reduction of the highly DETROIT, MICHIGAN ORSOND. BIRD RECEIVED AUGUST 21, 1939 potent Z-methyl-l,4-naphthoquinone.Since this preparation is soluble in dilute alkali and has a high degree of potency (approximately equal to ANTIHEMORRHAGIC ACTMTY OF SIMPLE the potency of 2-methyl-1,4-naphthoquinone),i t COMPOUNDS seems that this compound may prove very im- Sir: portant for intravenous vitamin K therapy. In connection with our investigation of vitamin Supplementing our previous preliminary re- K, we have tested recently a large number of port [THIS JOURNAL, 61, 1932 (1939)], we have derivatives of a-naphthoquinone, many of which reassayed several quinones to determine their have been prepared and reported by Professor optimum potency values. Except for 2-methyl- Fieser and his collaborators' and some of which 1,4-naphthoquinone, these more recent data agree were synthesized in this Laboratory. At this time with our previous findings. The assay of this we wish to report our findings of the antihemorcompound was carried out by the Thayer-Doisy rhagic activity of 2-methyl-l,4-naphthoquinone method, making a concurrent test, a t six, eighteen and of some other related substances of significance. and seventy-two hours. The standard was also One of the first quinones we assayed was 2run a t varying levels a t the same time for each methylnaphthoquinone and, because it did not series. The data are given in Table I. The de- appear a t that time to be as active as 2,3-ditails as to the method of feeding, care of the chicks, methyl-1,4-naphthoquinone,we did not deterlength of test period, manner of bleeding, etc., were mine its minimum dose. essentially the same as described previously. Following the appearance of the extremely interesting report of Ansbacher and Fernholz,2 we TABLE I reinvestigated the activity of 2-methyl-1,4-naphAssay period Thayer-Doisy units per mg. thoquinone, and we are now in complete agree6 hours 1110 18 hours 970 ment with them. 72 hours Average
1070 1050
( 1 ) Fieser, e l d.,THIS JOUENAL, 61, 1925, 1926, 2206 (1939). (2) Ansbacher and Pernh&, ibid., 61, 1924 (1980).
2564
COMMUNICATIONS TO THE EDITOR
Vol. 61
The assay method we employed was as follows: THE INTERCONVERSION OF CROTYL. ALCOHOL AND METHnVINYLCARBINOL IN AQUEOUS Day old chicks were placed on a vitamin K free SULFURIC ACID diet and kept on this diet for fourteen to seventeen Sir: days. When a preliminary determination of clotAccording to the generally accepted ideas ting time showed that a t least 90% of the chicks had clotting times above thirty minutes, the [Burton and Ingold, J . Chem. Soc., 1907 (1928); samples, dissolved in 0.1 cc. of peanut oil, were J. W. Baker, “Tautomerism,” pages 241-7, Routadministered orally. Eighteen hours later, the ledge, London, 19341, carbinol systems of the type H H H H H clotting times on the dosed birds were determined. R-C-C=CH R-C=C-COH Usually ten chicks were used a t each dose level. OH H H In the course of six weeks, nine assays were run are the least mobile of the allylic systems, exhibiton 2-methylnaphthoquinone. Following is a ing little or no tendency to isomerize. Although typical protocol of our results: the system has been reported to be mobile when Per cent. R = phenyl [Valeur and Luce, Bull. SOC. chim., 27, No. Clotting time in min. under Substance Dose birds 0-5 6-10 11-30 >30 10 min. 611 (1920)], Burton and Ingold [Zoc. cit.] were Z-Methyl-naphthounable to verify the observation. Another ex17 10 5 4 1 0 90 quinone ample of isomerization was reported by Prevost 2-Methyl-naphtho0.75y 9 4 3 1 1 77 qiiinone [Ann. chim., [lo] 10, 147 (1925)], who found that Z-Methyl-naphthocrotyl alcohol was formed during the dehydration quinone 0.5y 9 3 1 3 2 44 Alfalfa estract =75mg. 9 3 3 2 1 67 of methylvinylcarbinol over alumina at high Negative controls 0 10 0 0 0 10 0 temperatures. However, work on the mechanism of the reacIn all over 120 chicks were used in testing 2methyl-lj4-naphthoquinone a t levels of 0.5 to 1 . 0 ~ tion of the butenols with solutions of hydrogen and the results in every case were essentially the bromide [Young and Lane, THISJOURNAL, 60, 847 (1938)] convinced us that crotyl alcohol and same as above. From the results of Ansbacher and Fernholz methylvinylcarbinol should be interconvertible and from ours, i t would appear that, in the chick in the presence of acids to form equilibrium mixa t least, 2methylnaphthoquinone possesses anti- tures even a t room temperatures. This predicted hemorrhagic activity of the same order of magni- interconversion was based on a postulate that tude as the vitamin K1 reported by Thayer, et aLa activation of the oxonium ion of either crotyl alThese results are particularly striking inasmuch cohol or methylvinylcarbinol might lead to the as hitherto no simple compounds corresponding same resonating molecule H in chemical structure to the chemically identified HC CH vitamins exhibit the same order of activity as the i H ...H+ OH vitamins themselves. It is also noteworthy that HC-C’ 2-ethylnaphthoquinone (activity above 200 7 ) and HH 2-n-propylnaphth~quinone~ (inactive a t 400 7 )are in which the oxygen is bonded weakly to both decidedly less effective than the methyl homolog. carbons 1 and 3 [Young and Nozaki, paper in While 2,3-dimethylnaphthoquinone has some antiprocess of publication]. Consequently 50-ml. hemorrhagic activity (effective a t 50 r), 2,6- and portions of crotyl alcohol and methylvinylcar2,7-dimethyl-l ,4-naphthoquinone1 exhibit little, if binol are being treated with mixtures of water any (inactive a t 400 r), notwithstanding the fact and sulfuric acid adjusted so that the normality of that the substituents about the quinone systems of acid in 228 ml. of reaction mixture is 7.4, 3.7, and the latter two compounds are similar to 2-methyl1.9. With methylvinylcarbinol after one week naphthoquinone. a t room temperature the 7.4 N acid had caused RESEARCH LABORATORIES M. TISHLER MERCK& Co., INC. w.L. SAMPSON the production of 4.5 g. of crotyl alcohol and 3 g. of a fraction which appears to be a mixture of RAHWAY, NEWJERSEY, AND THE MERCKINSTITUTE OF crotyl and methylvinylcarbinyl ethers. With THERAPEUTIC RESEARCH crotyl alcohol after two weeks the 7.4 N acid had RECEIVED AUGUST18, 1939 produced 22 g. of the ether fraction and equal (3) Thayer, e l a!., Proc. SOC.E%$. Biol. M c d . , 41, 194 (1939). quantities, 4 g., of rnethylvinylcarbinol and (4) Prepared by Professor Fieier and his collaborators.
”/.I
