NO160873B - ACTUATOR FOR UNDERWATER LOCK VALVE WITH EXTERNAL MANUAL CONTROL AND ADJUSTMENT FOR STROEM. - Google Patents

Description

Process for the production of prolonged-acting ACTH preparations.

The invention relates to a method for production

of prolonged-acting ACTH preparations by reacting ACTH peptides in an aqueous medium with a phosphorus bridge-containing high molecular weight polymeric condensation products of phosphoric acid or thiophosphoric acid and an aromatic or aromatic-aliphatic compound containing two, three or more core-located OH, SH or NH groups which are either in the m- or p-position on the same core or are on different cores and which optionally also contain acidic sulfuric acid, phosphoric acid or thiophosphoric acid groups, and the method is characterized by the reaction taking place in the presence of native or partially degraded gelatin whose amino groups, possibly only partially, have been substituted by acylation or carboxy alkylation or which have become

treated with nitric acid by combining with each other

a) natural or synthetically produced aqueous solutions

ACTH peptides,

b) solutions of native or partially denatured gelatin, whose amino groups, possibly only partially, are substituted by

acylation or carboxy .-alkylation or which has been treated, with nitric acid and

c in aqueous solutions of the aforementioned phosphorus-containing condensation products

and optionally lyophilizes the resulting product.

As peptides with AGTH activity, it is suitable

besides the natural corticotropins with a chain length of 39 amino acids also synthetic peptides with a chain length of at least 20 amino acids, calculated from the amino end or with a chain length of at least 18 amino acids when the end-placed carboxyl groups are present as a^iid. In particular, peptides with a chain length of 23 - 24 amino acids are used, preferably as Cu-arnias, e.g. p 1-23J-corticotropin-2j-amide or |i^~^'<:>|"-corticotropin-24-amide,

the phthalin used according to the invention can be native or partially degraded. Degradation of the gelatin can be carried out, for example, via mild hydrolysis. This protein's free amino groups are completely or partially substituted, for example with acetyl, propionyl or acyl residues of higher aliphatic carboxylic acids. These acyl residues can in turn have hydroxyl or alkoxy groups. The acylation can also be carried out with acarboxylic acids, for example succinic acid, glutaric acid or adipic acid, with either the second carboxyl group present as an additional substituent or, during network formation, being bound to one of the protein's further amino groups.

Acyl residues can also include carbamoyl or carbalcoic acid residues. These carbamoyl or carbalcoic acid residues can be derived from aliphatic amines or diamines or aliphatic alcohols or diols. By aliphatic amines are preferably understood those with one to four carbon atoms, by aliphatic diamines preferably those with 2 to 6 carbon atoms. The same applies to the aliphatic alcohols or diols. Furthermore, the amino groups can be substituted with carboxyalkyl residues, for example carboxymethyl or carboxyethyl residues.

The acylation of the gelatin's free amino groups is carried out in the case of aliphatic carboxylic acid residues in the usual way, for example by reaction of carbo•; s/ruchlorido r c-n&Logt bermoylation of proteins, as aet f, ?k.~,. «i bc skr-, v . t x J,Viol. Cben.. I46 ( l<yiZ), sia a j' j L. Also" meil syrfjanhyc rid.? r it can acyleni.-., analogous to Aav.Prot. nhou»* ot ry j ' (I947}, stei a: ! 9?, Fs•<•*." to i ir..$c no :iv k<p>rbalkoxyderivftt^rie perform ss anul.-s-.o dot -.-on mentioned •» Naturs 1'3^ , (1935)) > oioo 4'j8 and in Biocne.v. j.-. (1(.), id $(><>>•> vod omsetnim; .i^o v\r»r • kuilsyreallcy V^t.-re. Ka^rw.o-, ;r'or..r; .:...-se ••>,5 :'r-.Mn3tilI' in; lykl.».>x bebt mei isoc;} tin ni. =>.r ell -r aiisocyanabfc '•. As niiko can ueb used : diifl^ty.1 --r: Lm ..-.ylfri-, • e.nraw rtylene- , p ,*nt «.methyl ?u- or nexamethylene-(f5.i.3y'.yc:i -'«t •' r. Cs le* we r 3 ts (clcb new •"■i-re slia:' c «1 ' " neabygg?aa) o"neU!.r.. •"'••. *< <; xJ.^ooyi-iH-.ber is '"ksK^p? '.vis be.i^rev^T, •

the German u.?t 1 rU . } ,~ l v' 71 c cv 1 > i v. ,1 _;.(.. For kår book ?. 1 <•/"'..-i'i f» oms ett si' man zn in' •r-v. cpet.'! an.viogt f re» the position w kar'o>ksy.n<?t.y'l collagen according to:; Ghw.. ubjtr. /;.'/' (l^j) <c>j01e mad hat^enka^xmsy aller t.ian till urar "••tert oe carbon }yr?:>- semi e.g. acrylic sypho toL the amino groups.

Disso iorbiudtjl.-">i"-r"-, aqueous solutions are aialyzed oc lyofil'.cjei^H et. tei end Lit i, week: ion niob '.' ostilled vanti.

Fr. 'ematiJ.ii.11g jn of deaminated gelatin (nat i v oller et cl-/i" degraded) by reaction mect nitric acid, which frig jcores from inorganic or organic ni tri t in acid solution is i and for se,;, known. Thus only num cjksemp". '.vis in acidic aqueous solution meil ote pH 1 and 4, preferably between pH 1.5 oa* 3/3 let MafJO,-, act on the gelatin wj<q> be e is et em p e ra c u r. Sulfuric acid is preferably used for acidification 3 n vend. Hydrochloric acid is less recommended as when it is used the deaminated gelatin easily becomes chlorinated. Once the reaction is finished, it is dialysed to remove salts against distilled water. The deaminated gelatin can be prepared by freeze-drying or by precipitation with a suitable solvent such as alcohol or acetone.Deoainero.s decomposed gelatin sa the careful decomposition is carried out advantageously according to the method indicated in the German patent No. 1.lio.792, ici et one heats at pH 6, 9 - 1.0 a few hours in an autoclave at 120°C, Desamirier no fore r. as then advantageously after acidifying the solution thus formed by means of sulfuric acid by adding NaM0o. A similar method is e.g. described in the German explanatory note rir. I.179.944.

If you want to work silk-free, you can also proceed in such a way that you mix the solution of gelatin in aqueous acetic acid with a nitric acid alkyl ester such as ethylene nitrite, butyl nitrite, amyl nitrite. The nitric acid released from the ester in an acidic environment then causes the deamination. After completion of the reaction, precipitate with alcohol or acetone; the precipitate is thoroughly digested with the same solvent, filtered off and dried in vacuum.

As higher molecular phosphoric acid esters are used, for example, the phosphoric acid esters of hydroxyl group-containing chalcone or dihydrochalcone derivatives, preferably polyphloretin phosphate. However, it is also generally possible to use higher molecular weight phosphorus bridge-containing condensation products as described in German patent no. I.024.677.

For the production of the new depot preparations, for example, the individual components are dissolved, e.g. an ACTH peptide, polyphloretin phosphate and one of the aforementioned gelatin derivatives separated in water, the acidic solution of the higher molecular weight phosphoric acid condensation product as well as possibly of the gelatin derivative being neutralized with an inorganic base such as KOH, NaOH or NH3, or an organic base, e.g. a tertiary amine such as triethylamine or N-ethyl-piperidine.

The filtered solutions of the additives are then combined in the desired order with the ACTH peptide solution. One adjusts with NaCl or a suitable physiologically tolerable organic compound, e.g. an amino acid, on isotonicity and corrects the pH of the solution to a value between 5.5 and 7»5> preferably 6.5 and J, 2. The solution is then filtered sterilely, optionally with the addition of bactericidal agents such as phenol, thymol and the like, filled up sterile and possibly lyophilized. If it is lyophilized, it is advantageous for the production of isotony to use monosodium glutamate.

The ratio of the individual components can be varied within certain limits. In general, phosphoric acid condensation products and gelatin derivatives are used in greater excess. Advantageously, however, at least the 50-fold weight amount calculated for ACTH peptide is used, the upper limit being 250 - 300-fold.

The depot effect of natural ACTH by adding, for example, polyphloretin phosphate is known from the German patent no. I.O24.677. By adding gelatin derivatives according to the invention, this depot effect of natural ACTH is clearly improved. Moreover, the particular advantage of the method according to the invention lies in the fact that not only the natural ACTH, but also synthetic ACTH peptides of shorter chain length can be transferred into depot preparations. It was previously not possible to transfer synthetic ACTH peptides in the manner known for natural ACTH in depot preparations of sufficient duration of action.

The method according to the invention has opposite it i

German patent no. 1,024*677 described method in addition to the improved depot effect also has the advantage that the process products in relation to the adducts known from this patent are in all cases easily soluble even at a neutral pH value, so that with regard to the injectability of the process products there is no limit-

nings.

From column 2 of the said patent, lines 46 to 50 appear

the fact that in certain cases and especially with low-molecular ACTH peptides, the coupling product can be poorly soluble at pH values around the neutral point. However, these embodiments can only refer to ACTH peptides containing at least 39 amino acids, as synthetic ACTH peptides of shorter sequence were not yet known at priority

the date of the German patent no. 1,024-677 • Now the known method is applied to synthetic ACTH peptides (approx. 18 to -24 amino acids,

i.e. with a considerably lower molecular weight) then, as was to be expected due to what is stated in the German patent no. 1,024,677 ae, the heavy solubility of the adducts formed is further increased.

It was now very surprising that after progress

the method according to the invention, both with synthetic and also with natural ACTH peptides, easily soluble adducts are always formed.

In relation to the state of the art, this means a clear technical progress

steps.

Particularly advantageously, the production of the ACTH preparations according to the invention is carried out with a prolonged effect so that first the ACTH peptide solution is combined with the gelatin derivative's solution

ning and the thus formed solution is mixed with the aqueous solution of higher molecular weight, a phosphorus bridge-containing polymer condensation products of phosphoric acid or thiophosphoric acid and an aromatic

or aromatic-aliphatic compound, containing three or more nuclear OH, SH or NH groups or only two such groups,

which are either located on different nuclei or in m- or p-

position on the same nucleus and which possibly also contain acidic sulfuric acid, phosphoric acid or thiophosphoric acid groups.

With this order of the mixing of the solutions, it is achieved that the eventual possible intermediate appearance of precipitation, which can only be avoided by longer stirring, can be avoided at the beginning, since all the mature gelatin derivative is present in excess that the easily soluble product of three components can immediately form .

The improved depot effect of the new AGT H-ada ukt is, which contains the gelatin derivatives used according to the invention 5. for-nold for depot effect ? i of ce previously known AOTH-polyphloretin-phosphate adducts and other owned lower son, depot-ACTil-recommended preparations such as e.g. gelatin itself and zinc complexes frevyar of f/Slgenae table. sou: we~ sr the corticosterone blood level in the peripheral blood in 1/100 ml vea ar^imatason-iortreated rats in l •'Spot of 1-6 hours after adm? preparation of the various depot preparations. In this context, "gelatin derivative 1" means a partially denatured and with nexame i-ylene diisocyanate newly formed gelatin, produced according to German patent no. 1,118,792, example 1.

"Gelatin derivative II" means a deaminated gelatin derivative, prepared according to example 3a.

The process products are valuable pharmaceuticals. They can e.g. used as a therapeutic for pituitary disorders, generally allergic reactions and diseases of the hematopoietic system. Example 1.

8.0 g of polyphloretin phosphate are dissolved in 50 ~ mJ of water. He neutralizes with 30 cm"^ l-n NaOH and filters. At the same time, 10 g of gelatin derivative I, (produced according to German patent no. I.II8.792, example 1 and subsequent lyophilization) is dissolved in 90 g of water; furthermore, 40 mS p<1 >~^-corticotropin-23-amide with 100 I.U./mg and 4OO mg NaCl in 20 cm^ of water. The solutions are brought together. After setting the pH to 6.8, top up with water to 200.0 cm^ and fill under sterile conditions in small bottles of each 1 cm-', lyophilize and seal.

Example 2.

8.0 g of polyphloretin phosphate are dissolved in 50 cm^' of water. Neutralize with 30 cm"^ l~n NaOH and filter. At the same time, 10 g of gelatin derivative I (prepared as in example 1) and 1.0 g of phenol are dissolved

in 90 cm~^ of water, the solution is neutralized with 2-n NaOH. Furthermore, 40 mg of p 1-2 '^-"> -corticotropin-23-amide is dissolved in 20 cmJ '■■> of water. The solutions are brought together after setting the pH to 7>0, filled with water to 200 .0 cm.3 and then filtered sterilely. One fills under sterile conditions into small bottles to every 1 cm-^ and seals. Example 3. a) Production of partially degraded, deaminated gelatin ("gelatin derivative II").

Dissolve 100 g of gelatin in 1 liter of water. The pH is set to 7 and heated in an autoclave for 5 hours at 120°C. After

cooling to room temperature (approx. 20°C) is adjusted with sulfuric acid to pH 2.5" Now add at approx. 20°C as long as an aqueous solution of NaNC^, until Iodkali-starch paper shows the presence of free nitric acid.

After 1 hour, it is cooled to 0° and dialyzed for 12 hours at this temperature against distilled water. By freeze-drying or pouring the aqueous solution into alcohol or acetone, the deaminated gelatin is obtained as a loose powder.

b) Dissolve 8.0 g of polyfluorene osf in 50 cm-^ of water. It is neutralized with 30 cm 3 l-n NaOH and filtered. At the same time, 10 g of gelatin derivative II (prepared according to point a)) is dissolved in 90 cm-1 of water; furthermore, 40 mg of p 1_pq-^-corticotropin-23-amide and 400 mg of NaCl are dissolved in 20 cm^ of water. The solutions are brought together. After setting the pH to 6.8, fill with water to 200.0 cm^, filter sterile, fill under sterile conditions into small bottles of 1 crn^, lyophilize and seal. Example 4.

8.0 g of polyphloretin phosphate are dissolved in 50 cm^ of water. One neutralizes mea 3° cm^ l-n NaOH and filters. At the same time, 10 g of gelatin derivative II (prepared according to example 3a) and 1.0 g of phenol are dissolved in 90 cm^ of water, the solution is neutralized with 2-n NaOH.

Furthermore, 40 mg of P""~<23_>lcorticotropin-23-amide is dissolved in 20 cm^ of water. The solutions are brought together, after setting the pH to 7»0, filled with water to 200.0 cm^ and filtered sterilely. fulfill under sterile conditions in small bottles of 1 crn-^ and seal Example 5.

Dissolve 8.0 g of polyphloretin phosphate in 50 cm^ of water. Neutralize with 30 cm^ of NaOH and filter. At the same time, dissolve 10 g of gelatin derivative I (prepared as in example 1), 40 mg of 3-corticotropin-23-amide with 100 I.E./mg and 400 mg NaCl in 100 cm^ water. To this solution, the polyphloretin phosphate solution is allowed to flow slowly while stirring. A clear to slightly opalescent solution is obtained. After setting the pH to 6.5 - 7, top up with water to 200.0 cm^, filter sterile and fill under sterile conditions into small bottles of 1 - 2 cm^, lyophilize and seal Example 6.

Dissolve 8.0 g of polyflourine osf in 50 cm-^ of water. It is neutralized with 30 cm 3 l-n NaOH. At the same time, 10 g of gelatin derivative I (prepared as in example 1) dissolve 40 mg of ^""""^-corticotropin-

23-amide with 100 I.E./mg and 400 mg NaCl in 100 cm^ of water. The polyphloretin phosphate solution is slowly added to this solution while stirring. You get a clear to slightly opalescent solution. After setting the pH to 6.5 - 7, it is filled with water to 200.0 cm^ and sterile filtered. It is filled under sterile conditions into small bottles every 1-2 cm^ and sealed.

Example 7.

One works as in example 5", but instead of the gelatin derivative described there, 10 g of carboxymethyl gelatin, prepared according to Chem. Abstract 47, (1953), page 901e.

Example 8.

One works as indicated in example 5", however, instead of the gelatin derivative described there, 10 g of gelatin derivative II, prepared as indicated in example 3a" and treated with nitric acid is used.

Example 9.

Dissolve 6.0 g of polyphloretin phosphate in 50 cm^ of water. It is neutralized with 22.5 cm 3 l-n NaOH and filtered. -At the same time, dissolve 7"5 S gelatin derivative I (prepared as stated in example 1), 40 mg of β1"2^-corticotropin-23-amide with 100 I.U./mg and 2.5 sodium glutamate in 100 cm^ of water. To this solution, the polyphloretin phosphate solution is allowed to flow slowly while stirring. A clear to slightly opalescent solution is obtained. After setting the pH to 6>5 - 7.0, fill with water to 200.0 cm^, filter sterilely and fill under sterile conditions in bottles to 1-2 cm^, lyophilize and seal.

Claims (1)

Process for the production of prolonged-acting ACTH preparations by reacting ACTH peptides in an aqueous medium with a phosphorus bridge-containing high molecular weight polymeric condensation products of phosphoric acid or thiophosphoric acid and an aromatic or aromatic-aliphatic compound containing two, three or more core-placed OH, SH- or NH groups which are either in the m- or p-position on the same nucleus or are located on different nuclei and which possibly also contain acidic sulfuric acid, phosphoric acid or thiophosphoric acid groups, characterized in that the reaction takes place in the presence of native or partially degraded gelatin whose amino groups, possibly only partially, are substituted by acylation or carboxy-alkylation or which has been treated with nitric acid, by combining with each other a) aqueous solutions of natural or synthetically produced ACTH peptides, b) solutions of native or partially degraded gelatin, whose amino groups, possibly only partially, are substituted by acylation or carboxy alkylation or which has been treated with nitric acid and c) aqueous solutions of the aforementioned phosphorus-containing condensates products, and optionally lyophilizes the product thus formed.