Patented May 2, 1950 2,506,294 UNITED STATES PATENT OFFICE 2,506,294 ADRENOCHROME MONO - SEMICARBAZONE COMPOUND AND HAEMOSTATIC COMPO STON Georges Dechamps, Liege, Henri Le Bihan, Em bourg, and Camille Beaudet, Liege, Belgium, assignors to Societe Belge de Azote et des Produits Chimiques du Marly, Liege, Belgium, a corporation of Belgium No Drawing. Application May 2, 1947, Serial No. 749,592. In Belgium November 29, 1943 Section 1, Public Law 690, August 8, 1946 Patent expires November 29, 1963 2 Claims. (C. 6-65) 2 The invention relates to a process for the prep Solutions in Water or in other solvents, such as aration of a stable haemostatic derivative of ethanol, even very dilute, are also very stable. adrenochrome and to the products obtained Notwithstanding their relatively great chemical thereby. More particularly, it pertains to the inactivity, they retain in a large measure the val preparation of products which result from reac luable haemostatic properties of adrenochrome tion between adrenochrome and a salt of a ke Solutions. This important property has been tone reactive amine derivative as the Salts of hy used for therapeutic purpose by us for prepar droxylamine, semicarbazide, phenylhydrazine ing stable haemostatic solutions which may be and nitrophenylhydrazines, and includes corre kept in ampules. lated improvements and discoveries whereby the 10 Clinical experimentation with the various aforesaid derivatives may be effectively prepared. hereinbefore mentioned products has shown that It is known that some oxidation products of the intensity and the specificity of the haemo adrenalin have a pharmacodynamic activity, es static action differ in a rather unexpected man pecially a haemostatic activity similar to that of ner from one product to the other. adrenalin itself or of its derivatives, Such as 5 In the haemostatic medication with the human adrenalone. Among these oxidation products, being, the adrenochrome semicarbazone is par the adrenochrome or N-methyl-3-hydroxy-2,3- ticularly suitable. The therapeutic effects of this dihydro-5.6-quinoneindol is remarkable for its latter compound are so similar to adrenochrome efficiency as a haemostatic agent at very Small that it may be considered as a stabilized form of doses (for instance, about 106 gr. With the rab 20 adrenochrome with the therapeutic applications bit) and for its more rapid and equally intense of the latter being thereby made possible. action than that of adrenalin. Compared to The practical examples given hereinafter illus the latter, adrenochrome presents even the ad trate the application of the invention by means vantage that it does not alter the cardiac rhythm of the adrenochrome semicarbazone, but it is to and does not cause any hypertension or internal 25 be understood that the invention is not limited to haemorrhages. Thanks to these properties, this particular application. By modifying the adrenochrome should be suitable for many ther method of treatment and the proportions of the apeutic applications, for instance in Surgical Op reagents used, according to the nature of these erations, in the treatment of internal haemor reagents, it is possible to prepare, in a similar rhages, etc. Unfortunately its instability, in 30 manner and Within the scope of the invention, aqueous or alcoholic solution, makes its use Sub the corresponding haemostatic agents of adreno stantially impossible in the medical art. chrome monoxime, monophenylhydrazone and Adrenochrome decomposes in 35 minutes in an mononitrophenylhydrazones. aqueous solution of pH 7.3; in 4 minutes in a 1% acid solution of hydrochloric acid, and in 40 sec i Eacample 1 onds in a 1% ammonia solution. This instability of adrenochrome is principally attributed to the A suspension containing 1 part by weight of quinone function of the benzene ring of its mole adrenalin and 2 to 6 parts by weight of silver cule. Therefore, we have tried to stabilize the oxide in 150 to 250 parts by weight of methanol quinone function. 40 or ethanol is stirred for about 10 minutes. The Considering adrenochrome as a diketone, We alcoholic adrenochrome Solution obtained is sep have used, for this purpose, certain ketone reac arated by draining and the filtrate is quickly tive reagents, more particularly amine derivatives evaporated to dryness at low temperature and in such as salts of hydroxylamine, semicarbazide, Vacuo. The red crystals of adrenochrome ob phenylhydrazine and various nitrophenylhydra tained are dissolved in 45 to 55 parts by weight of zines. All these reagents condense easily with water. To this solution, 2 parts of sodium ace adrenochrome and form by reaction with one tate dissolved in 2 to 3 parts of water and 2 parts oxygen atom of the quinone function, the corre of Semicarbazide hydrochloride dissolved in 2 to sponding monoxime, Semicarbazone, phenylhy 3 parts of Water are added. The formed pre drazone and nitrophenylhydrazones. These Con 50 cipitate consisting of red-orange prismatic nee densation products which are much less soluble diles is separated by filtration and recrystallized than adrenochrome and easy to obtain, in pure from diluted ethanol. There is obtained 0.30 to state, in the form of crystals, are much more 0.40 part by weight of adrenochrome mono-semi stable as to spontaneous transformation and de carbazone dihydrate, melting at 203 C. with de composition than adrenochrone itself. Their composition. 2,506, 294 - 3 4. Eacample 2 occurrence. Also, the solutions of semi-carba Zone of adrenochrome, which even in strong doses 10 parts by weight of laevorotatory adrenalin do not show any secondary harmful or toxic are dissolved in a mixture of 220 parts of pure" action, constitute important progress in the pre methanol and 6, 6 parts of formic acid of 85% vention of hemorrhages arising in surgical oper strength. To this mixture, while-heated to 35°C., ations and/or in the treatment of pathological or 40 parts of silver oxide are added under cons accidental bleeding. tinuous stirring. After one minute's stirring, the As illustrative of clinical application; when liquid is rapidly separated by filtration and mains injected by way of a preventive from 15 to 60 tained at -20° C., for twenty minutes. They minutes before any surgical operation in a dose of Crude adrenochrone thus obtained is filtere one or more 100-gamma, ampules, solutions of the washed with 20 parts of methanol, at 0°C., and: semi-carbazone of adrenochrome have had the recrystallized at 35-40° C. from a mixture of 39 effect of reducing greatly the intensity as well parts of methanol and 0.48 part of formic acid of: as the duration of hemorrhages which would 85% strength. After cooling, the crystals: are: 15 normally be produced, for example, in pros filtered under vacuum. 3.4 gr. of dark-red crys tatectomy, tonsillectomy, tals of pure adrenochrome are: thus obtained, Operations on the which are dissolved in 275 parts of water. To this nervous System, etc. Solution is added a solution of 5.4 parts of semi By injection as a curative, during hemorrhages, carbazide hydrochloride' and 5.4 parts of sodium and in a dose of one or more 500-gamma ampules, acetate, in 34 parts of water. The formed pre- 20 post-operative bleeding or spontaneous hemor cipitate is filtered under vacuum. and recrystal. rhages of the digestive tract and of the respira lized from diluted ethyl alcohol. 3.7 parts, of: tory organs have furthermore. been arrested. orange-red adrenochrome mono-semicarbazone It Will be understood that in order to prepare, dihydrate having a melting point of 203 C., with. according to the invention, a stable. haemostatic decomposition, are thus, obtained. adrenochrome, derivative in the form of mono Aqueous solutions. of the semi-carbazones of Semicarbazone, it is not necessary, as in the above: adrenochrome, even very dilute, are perfectly. described example, to start from a product ob stable, keep indefinitely in sealed ampules, and tained from adrenalin, by...means of silver oxide. may be. used for subcutaneous or intramuscular Adrenochrome, an unstable. and intermediate injections. To this end, isotonic solutions are: 0 compound. between adrenalin and adrenochrome: advantageously. prepaired, adjusted to a pH in. mono-Semicarbazone, may be prepared by any the neighborhood of 7. and which may be em other method, for instance, by diastasic; oxida ployed as a preventive or as a curative according. tion of adrenalin, to the dose. of semi-carbazone: administered and What we claim is: according to whether this, administration is given. 35, 1. As a compositions of matter, an adreno before. Or during a hemorrhage. chrone semicarbazone. In the preventive capacity, that is...to, say, for 2. As a composition of matter, a stable in the purpose of avoiding or attenuating the hem-- jectable haemoStatic isotonic aqueous. solution. orrhages.normally produced in surgical opera containing adreriochrome mono-semicarbazone: tions, use is preferably made of Solutions of a rel-. 40 GEORGES DECHAMPS. atively low concentration, for example, at one HENRILEBHAN 1. ten-thousandth . (1:10000), i.e. 100 gammas (0.1. CAMILLE BEAUDET mg.) per cc. A dose. of, for example, 1.cc., of a. -- Solution of 100 gammas, per-cc. injected a certains 45. REFERENCES. CITED time (for example; 4 to 1 hour), before an oper ation, usually bloody, allows, of reducing, the. The following references are of record in the: bleeding in great proportions, and - in certain. file of this patent: cases, even practically suppressing it and/or...re-. UNITED STATES PATENTS. ducing its; duration. 50. Number Nanae: Date. Utilized in: a stronger... dose, such as one. can: 2,162,863. Ripke ------June 20, 1939. realize by injection, one or more times, of... a. larger volume of solution-of.a.low.concentration, OTHER FREFERENCES for exa:nple, 100 gammaS per cc. (a gamma is a Deronaux: Archives Internationales... de Phar unit of weight equalito, the one millionth part of macodynamie et de Therapie, pages. 202-219. a gram, 0.000,000,353 ounce; Hackh's Chemical i. Squibb. Abstract Bull., vol. 18. (1945), page 1557. Dictionary 1929, page 322), or preferably by the Veer: Rec. Trav. Chimde. Pays-Bas, vol. 61. use of more concentrated solutions, of for exam. (1942), pages. 638 to. 646. ple, 500 gammaS per CC, or more, the semiscarbas Chem. Abstracts. 40: 947 (Feb. 20, 1946), citing Zone of adrenochrome, is very eficacious also for Compt. Rend. Soc. Biol., vol. 38 (1944), pages retarding: and arresting hemorrhages: in... their 858,