March 15, 1960 E. R. DiczFALUsy ETAL 2,928,849 HIGH-MOLECULAR WEIGHT DERWATIVES OF STEROIDS CONTAINING Filed Nov. 8, 1954HYDROXYL GROUPS AND METHOD OF PRODUCING THE2. SAMESheets-Sheet 2 S. s S s SN N g Q R S SS N. S. mN SSS SNS s SS S Q S R sy R.S N- SS N y g s N & s S h N h (N S 2 SS S S. SS S. SSr S. S s s S V S -- < S n -1 q S. -1 N SR & \ 1 -1 S2 S \/y SN S s SS / 9 S W. AS / & / (C N R / lu / R.Ska. N 9 N. / SS ( A / 8S Vy N - N S$ < + c & o, N NS N N Y Xnu N. N SS N s VO N v INVENTORS Egow Artaarar of CzaaZ Sy owa anar Gear afaravo Ayaws vangga aex AAwu7 sear7 AyoGeAG o N er s 7oarszaw owe aavou favoa aro swww.pittu w v? V2/1soya 7vy 1 mga volholzm Y Á4rl-47243, Attorwey, United States Patent Office 2,928,849 rest swt, ... arts. Patiented Mar. 5, 1960 2 2,928,849 whether a sufficiently even and continuous resorption can HIGH-MOLECULAR WEIGHT DERVATIVES OF be obtained in this manner. Moreover, the implanted STEROIDS CONTAINING HYDROXYL GROUP8 object is often embarrassing, and in some cases it will AND METHOD OF PRODUCING THE SAME be expelled from the place of implantation. In respect Egon Rihardt Diczfalusy, Solentuna, and Ove Birger 5 of the preparations mentioned under (b) and (c) it will Fernö, Hans Jakob Fex, Knut Bertill Högberg, and further be difficult to insure perfect sterility. Torsten Ove Enok Linderot, Hasingborg, Sweden, as It is an object of the present invention to prepare high signors to Aktiebolaget Leo, Haisingborg, Sweden, a molecular weight compounds of hydroxyl group-contain firm ing steroids, from which compounds it is possible to 10 make preparations for therapeutic purpose which do not Application November 8, 1954, Serial No. 467,482 have the disadvantages of previous preparations, and in which the effect of the hormone, when the preparation Claims priority, application Denmark November 20, 1953 is administered, is prolonged. in many cases, greatly 2 Claims. (Ci. 260-397.5) prolonged effectiveness of hormonal action is secured. 5 The high-molecular compounds, according to the pres This invention relates to high-molecular derivatives of ent invention, are esters of the acids of phosphorus (in hydroxyl group-containing steroids, i.e., steroids with cluding phosphorus and thiophosphoric acid) with hy hydroxyl groups in the molecules as well as steroids in droxyl group-containing steroids (including steroids in which the hydroxyl groups result from the enolization of which the hydroxyl groups result from the enolization of keto groups. Compounds of both types are intended to keto groups). They consist of chain-shaped molecules, be included within the scope of the term "steroids con the single links of which are coupled together by the taining free hydroxyl groups,' or "hydroxyl group-con phosphorus acid groups, and in which the steroid is pres taining steroids,' as used herein. ent in the form of steroid-phosphorus acid groups. This invention has for its especial object the produc Here and later on in the specification and in the ap tion of certain derivatives which are capable of produc 5 ing and maintaining, in an organism, the biological ef 2 pended claims the term "phosphorus acid groups' should fects of the steroid contained in that derivative for a be understood as including phosphate as well as phosphite longer period of time than has hitherto been possible. and thiophosphate groups, and the term "chain-shaped The invention is of particular interest in connection with molecules' should be understood as comprising both those steroids having hormonal effects. straight chain and branched chain molecules. It is known that, at any particular moment of time, the In the disclosure which follows a detailed explanation hormone content in a hormone-producing organ is low will be given as to the nature of the novel high-molecular in comparison with the amount of hormone which must weight compounds and the kind of groups contained be supplied to that organism in order to obtain a distinct therein, as well as details regarding a preferred method hormonal effect. This has been clearly established by for producing them. The novel method by which the animal tests, as well as by the clinical use of the hor compounds can be prepared is also a specific object of mones in question. The conclusion can be drawn from this invention. Further, the effects which can be ob this that the hormones are continuously excreted from tained by means of the novel compounds upon their ad the seat at which they are produced, and that they do iministrationexamples. will be illustrated by means of typical not accumulate therein. Accordingly, the best way of 4. The high-molecular compounds of the invention can administering hormones should be to imitate this natural be produced by reacting the steroid with a phosphorylat biologic state of affairs insofar as possible, and to this ing agent, this term being herein used to include thio end it is a primary purpose of this invention to produce phosphorylating agents, under such conditions in respect ihormone preparations having continuous and protracted of the amount and concentration of the phosphorylating effect. agent, the reaction temperature and the time elapsing be The previous attempts to prepare steroid hormone 45 fore the reaction is stopped, that the steroid molecules preparations having protracted effects may be summed up alone or together with a coupling substance form high :as follows: molecular products in which the steroid is contained in (a) Administration of derivatives, mainly esters, as the shape of steroid-phosphorus acid groups. 'for instance testosterone propionate and estradiol ben By varying the conditions under which the phos Zdate, where the prolonged effect is produced thereby 50 phorylation takes place, products may be obtained, hav ithat the substance has to be hydrolyzed in the organism ing different molecular sizes, whereby it is possible to ibefore a biological effect can set in. get a more or less protracted effect of the compounds. (b) Administration of the hormone in oil depots Generally, the reaction is expediently carried through wherefrom it is only slowly resorbed. at temperatures below 0° C., but for tardy reactions it By combining (a) and (b), preparations can be pro 55 may be expedient to increase the reaction temperature .duced which combine both principles. up to the neighborhood of 100 C., if desired. (c) Administration of the hormone in solid amor According to the invention it is expedient to continue phous form as a suspension or an adsorbate on, for ex the treatment with the phosphorylating agent until the ample, carbon or aluminum phosphate, by injecting a molecular weight of the products is more than 2000, suspension of micro-crystals or by implanting tablets of 30 since this will result in products that exhibit a com imicro-crystals, the so-called pellets. pletely satisfying protracted effect. ... None of the said methods are particularly satisfying. When the reaction has reached a suitable stage which, Thus injection of the hormone in oil must be repeated for instance, may be controlled by testing the dialyz. fairly often, which means that unpleasant oil depots are ability of the product, the reaction is stopped. Accord left which the organism cannot dispose of at all, or can 65 ing to the invention this may expediently be accom only, dispose of with difficulty. Preparations containing plished by hydrolysis of the reaction mixture. Thus, for steroid hormones adsorbed on, for example, aluminum instance, the reaction may be stopped by adding crushed phosphate produce inflamatory reactions and other sec ice to the reaction mixture, or by pouring the latter into sondary effects. By implantation of the so-called pellets, ice water. " . an uneven and discontinuous progress of the resorption According to a particularly satisfactory method of car process is often noticed, and all in all it is questionable rying out the invention, the steroid is treated with an - 2,928,849 . 3 4. amount of phosphorylating agent not substantially ex thiophosphorylation. Examples of such substances are ceeding the equimolar amount, and in case a coupling polyvinyl alcohol, dextran, gelatin and other similar substance is further used it is preferable that the phos high-molecular weight products which are compatible phorylating agent is used in an amount not substantially with the organism. -, - exceeding the equimolar amount as compared with the The use of a coupling substance makes it further pos total amount of steroid and coupling substance. sible to build up high-molecular compounds from steroids By limiting the amount of phosphorylating agent in this having in the molecule only one hydroxyl group including manner, the formation of chain-shaped molecules is pro such hydroxyl group which results from a keto group moted, since the single molecules of the phosphorylating being enolized, whereas, if no coupling substance is used, agent thereby have greater possibilities of sinultaneously O the steroid in question should have at least two such reacting with different molecules of the steroid, the cou groups in the molecule in order that the chain formation pling substance, or with both. necessary for producing high-molecular weight com As it appears from this description of the method, it pounds may be attained. is a condition for its accomplishment that the steroid According to a preferred embodiment of the inven contains one or more hydroxyl groups including hydroxyl 15 tion a steroid having in the molecule only one hydroxyl groups resulting from keto groups being enolized, since group including such hydroxyl group which results from steroids not containing such groups cannot be phos a keto group being enolized is coupled by phosphoryla phorylated.