DE1240223B - Process for the production of protracted ACTH preparations - Google Patents

Description

GERMAN WTTWt PATENT OFFICE DeutscheKl .: 30 h -2/10

EDITORIAL

Number: 1 240 223

File number: F 44686IV a / 30 h

^ 240 223 Filing date: December 12, 1964

Open date: May 11, 1967

The reaction of ACTH with oxypolygelatine is known from German patent specification 951228. Hereby a potentiation of the ACTH effect is achieved, which results in a sudden strong one Expresses a reaction immediately after the injection of the preparation.

From the German patent specification 960579 it is known that one can be sterilized in the heat ACTH preparation is obtained when ACTH is reacted with non-antigenic gelatin in an aqueous solution. In this patent mentions that ACTH is highly insoluble in gelatin and therefore has a prolonged physiological activity. Furthermore, it is described in the patent mentioned, that by adding a "delaying agent" a prolonged therapeutic Effect achieved. As such retarding agents, sparingly water-soluble astringents are used called, so that it could be seen from this patent that a prolonged physiological activity can only be achieved through the poor solubility of ACTH in the medium in question.

The German patent specification 920 148 describes a process for the production of ACTH preparations with a prolonged duration of action. Such preparations are obtained by adding an alkaline reacting protein or protein degradation product and a water-soluble zinc salt to an aqueous ACTH solution. According to this patent specification, the prolonged action of ACTH is achieved by the added substances, since this creates a sparingly soluble compound which forms a non-coagulating suspension in an aqueous medium.

It has now surprisingly been found that easily soluble ACTH preparations with a protracted action can be obtained from ACTH-effective peptides and higher molecular weight, containing a phosphorus bridge polymeric condensation products of phosphoric acid or thiophosphoric acid and an aromatic or aromatic-aliphatic compound which has three or more nuclear OH, SH or NH groups or only two such groups, either on different nuclei or in m- or p-position are located on the same nucleus, and the possibly still acidic sulfuric acid, phosphoric acid or contain thiophosphoric acid groups, with the formation of adducts in aqueous solution when you get

a) the said phosphorus-containing condensation products,

b) natural or synthetic ACTH peptides and

Process for the production of protracted
acting ACTH preparations

Applicant:

Farbwerke Hoechst Aktiengesellschaft

formerly Master Lucius & Brüning, Frankfurt / M.

Named as inventor:

Dr. Gerhard Vogel, Hofheim (Taunus);

Dr. Rolf Geiger, Frankfurt / M .;

Dr. Karl Sturm,

Frankfurt / M.-Unterliederbach

c) native or partially degraded gelatin, the amino groups of which, if necessary only partially, are substituted by acylation or carboxyalkylation,

combines in aqueous solution and the product is optionally lyophilized.

In addition to the natural corticotropins with a chain length of 39 amino acids, synthetic peptides with a chain length of at least 20 amino acids, counting from the amino end, or with a chain length of at least 18 amino acids if the terminal carboxyl group is an amide are suitable as peptides with ACTH activity . In particular, peptides with a chain length of 23 to 24 amino acids, preferably as ω-amides, are used, e.g. B. the ß ^ -Cor ticotropin-23-amide or the / 3 ¹ - ²⁴ -Corticotropin-24-amide.

Examples of higher molecular weight phosphoric acid esters are those in the German patent 929 664 described phosphoric acid esters of hydroxyl-containing chalcone or dihydrochalkone derivatives, preferably polyphloretin phosphate, in question. However, in general, higher molecular weight, Condensation products containing phosphorus bridges, as described in the German patent 1024 677 can be used.

709 579/391

1 240

223

The gelatin used according to the invention for the additional component of the adduct which is physiologically compatible can be used natively, for. B. an amino acid, isotonic and grain or partially degraded. The degradation of the gela- gated the pH of the solution to a value between tine can z. B. made by mild hydrolysis 5.5 and 7.5, preferably 6.5 and 7.2. The solution will be men. The free amino groups of this product 5 is then sterile filtered, sterile filled and gegebenenteins are completely or partially substituted beispiels- if lyophilized.

wise by acetyl, propionyl or acyl radicals The ratio of the individual components can

higher aliphatic carboxylic acids. These acyl radicals can be varied within certain limits. Generally

can in turn be hydroxyl or alkoxy groups and phosphoric acid condensation product

wear. The acylation can also be used with dicarboxylic io gelatin derivative in greater excess,

ren, for example succinic acid, glutaric acid or is advantageous in each case at least 50 times

Adipic acid, be carried out, either amount by weight, based on ACTH peptide, ver

uses the second carboxyl group as an additional substitute, the upper limit is 250-

tuent is present or linked to another up to 300 times.

Amino group of the protein is bound. 15 The surprising effect of the invention as acyl and carbamoyl or procedure are located particularly in the fact that well-carbalkoxy wasserlös in question. These carbamoyl or ACTH adducts are produced, which carbalkoxy radicals can be derivatives of aliphatic, despite their easy solubility, show a protracted appearance of amines or diamines or aliphatic kung. This result has to be in terms of alcohols or diols. Completely surprising among aliphatic prior art.
Amines are preferably those with 1 to 4 carbon atoms. On the other hand, this is also where the special carbon atoms, among aliphatic diamines, the progressiveness of the process according to the invention lies: preferably those with 2 to 6 carbon atoms. The same applies to the aliphatic tentschrift 920 148, which gives poorly soluble Injek alcohols or diols. Furthermore, solutions containing amino tion 35th The disadvantages and additional measures will be substituted by the easy water solubility of groups by carboxyalkyl radicals, for example the ACTH preparations obtained according to the invention, carboxymethyl or carboxyethyl radicals. the-The Injektionspräpa acylation of the free amino groups of the overall rat with a water-insoluble may be required, and thus his hand Latine 30 may dling difficult (cf. in the case of the aliphatic carboxylic acid. German Patent residues in the customary manner, for example by environmental 920148 , P. 2, line 115, to p. 3, line 16), Ver setting with carboxylic acid chlorides or anhydrides, avoided.

be carried out, for example analogously to that in The method according to the invention has against-J. Biochem. Chem., 146 (1942), p. 331, descriptive over the process of the German patent NEN benzoylation of proteins. The implementation 35 920 148, however, still has another advantage. The development of ACTH preparations described from gelatin with acid anhydrides is analogous to the method known from this patent specification for the preparation of ACTH preparations. natural ACTH peptides, while it is not applicable when using the production of carbalkoxy derivatives from synthetic ACTH peptides from about 20 to a known manner by reacting with 40 to 24 amino acids, which are made at the time-chlorocarbonic acid alkyl esters, as made at the point of application of the cited patent, for example, from Nature, 136 (1935), p. 438, or were not known.

from Biochem. Journ., 33, p. 908 (1939). The depot effect of natural ACTH

The preparation of the carbamoyl compounds by adding, for example, polyphloretin phosphate

works best with isocyanates or diisocyanates. 45 phat is from the German patent specification 1024 677

As such come into question: Dimethylene, tri-known. By the addition of the invention

methylene, tetramethylene, pentamethylene or gelatin derivatives to form new adducts is the hitherto

Hexamethylene diisocyanate. Implementation of the well-known depot effect of natural ACTH

Latin (native or partially degraded) with Diisocyana- significantly improved. In addition, the special

ten is, for example, in the German patent documents 5 ° more advantage of the method according to the invention

th 1,118,792 and 1,155,134. Go to carb- in that not only natural ACTH, but also

Oxyalkylation puts the amino groups in with synthetic ACTH peptides of shorter chain length

Halocarboxylic acids to, or you can store unseeded depot preparations can be transferred. It

saturated carboxylic acids, such as. B. Acrylic acid, which was previously not possible to use synthetic ACTH-Pep-

Amino groups. 55 tide in the manner known for natural ACTH

For the production of the new depot preparations dissolves in depot preparations of sufficient effectiveness

For example, you can transfer the individual components, e.g. permanently.

an ACTH peptide, polyphloretin phosphate and the beneficial influence of the gelatin derivatives on the gelatin derivatives mentioned, separated in water, the depot effect of the ACTH polyphloretin AdWith the acidic solution of the higher molecular weight phosphorus 60 products can be seen from the following table, which phosphoric acid -Condensation product as well as given corticosterone blood level in the peripheral blood, at least that of the gelatin derivative with an inorganic base pretreated with dexamethasone, such as KOH, NaOH or NH ₃ , or within 1 to 6 hours after administration of an organic one Base, e.g. B. a tertiary amine, which shows various depot preparations. In such a way as triethylamine or N-ethyl-piperidine, neutral- 65 this table is divided under "gelatin derivative" a. while degraded and with hexamethylene diisocyanate. The filtered solutions are then crosslinked gelatine, manufactured according to German unity. One sets with NaCl or a suitable, patent 1118 792, example 1, understood.

Claims (1)

5 6 Depot preparation 1 ² i
hours ⁴ I. 6th Natural ACTH + polyphloretin phosphate 48 47 I.
35 i 10 Natural ACTH + gelatin derivative 51 47 42 I. 25th / Ji-23 -Corticotropin-23-amide + polyphloretin phosphate 45 37 14th 6th y3i -23 _Corticotropin-23-amide + gelatin derivative 49 46 41 y 21 The products of the process are valuable pharmaceuticals. You can e.g. B. as therapeutic agents for pituitary disorders, general allergic reactions or diseases of the haematopoietic system be used. example 1 8.0 g of polyphloretin phosphate are dissolved in 50 ecm of water. It is neutralized with 30 ecm in NaOH and filtered. At the same time, 10 g of gelatin derivative (prepared according to German Patent 1118 792, Example 1, and then lyophilized) are dissolved in 90 ecm of water; Furthermore, 40 mg / J ¹ - ² S -corticotropin-23-amide is dissolved with 100 IU / mg and 400 mg NaCl in 20 ecm of water. The solutions are combined. After the pH has been adjusted to 6.8, the mixture is made up to 200.0 ecm with water, filtered under sterile conditions and filled into vials of 1 ecm each under sterile conditions, lyophilized and sealed. Example 2 _{3 <J} 8.0 g of polyphloretin phosphate are dissolved in 50 ecm of water. It is neutralized with 30 ecm in NaOH and filtered. At the same time, 10 g of gelatin derivative (prepared according to German Patent 1118 792, Example 1, and then lyophilized) and 1.0 g of phenol are dissolved in 90 ecm of water and the solution is neutralized with 2N NaOH. Further, dissolving 40 mg / ^{i: 1} - ²³ -Corticotropin-23-amide in 20 cc of water. The solutions are combined, after the pH has been adjusted to 7.0, the mixture is made up to 200.0 ecm with water and sterile filtered. Vials of 1 ecm each are filled under sterile conditions and sealed. Claim: Process for the production of protracted ACTH preparations from ACTH-effective Peptides and higher molecular weight polymeric condensation products containing a phosphorus bridge from phosphoric acid or thiophosphoric acid and an aromatic or aromatic-aliphatic Compound that has three or more nuclear OH, SH or NH groups or only two such groups that are either are located on different nuclei or in the m- or p-position on the same nucleus, and which may be still contain acidic sulfuric acid, phosphoric acid or thiophosphoric acid groups, with formation of adducts in aqueous solution, characterized in that one a) the said phosphorus-containing condensation products, b) natural or synthetic ACTH peptides and c) native or partially degraded gelatin, their amino groups, optionally only are partially substituted by acylation or carboxyalkylation, combines in aqueous solution and the product is optionally lyophilized. Considered publications:
German patents No. 1118 792, 1155 134,; i 228, 960 579, 920148.