DE2338970A1 - Standardised, stabilised, water-soluble placenta extracts - used in the treatment of wounds and swellings e.g. ulcers - Google Patents

Abstract

A placenta extract obtd. in known ways using solvents and centrifugation, is freed from hormones and residual lipoids by addn. of further organic solvents, which are subsequently removed. Proteins are then removed by addn. of preserving agent at pH 7 and acidification with organic acids to pH 6.5-7.4. A soln. contg. customary buffers and preserving agents as well as a series of low-molecular components known to be present in the placenta, is then added to stabilize the prodt. (I). (I) are used in the treatment of wounds and swelling such as ulcers, as well as a component in cosmetic prepns. The contents of the extracts may be regulated without changing its principles. (I) may be stored for years and has better activity of longer duration than known extracts.

Description

PATENT Attorney DIPL.-ING. WOLFGANG BRINGMANN

4 Düsseldorf 11, Rheinallee 130

Böttger limited partnership
Pharmaceutical and Cosmetic
preparations

1000 Berlin 31
Paulsborner Strasse 2

Process for the production of standardized, stabilized, water-soluble placenta extracts.

It is known to use water-soluble placenta extracts both for the treatment of wounds and ulcers, e.g. ulcers as well to be used as a mixed ingredient in cosmetic preparations. When treating with water-soluble placenta extracts However, significant fluctuations in effect occur, which include can also be attributed to changes in the composition of the extracts produced.

The invention is therefore based on the object of providing a process for the production of the extracts mentioned at the outset, whose composition can be regulated without, however, changing or losing their basis, i.e. that all the benefits and effects of natural, water-soluble placenta extracts are retained.

After extensive investigations it was found that the treatment of placentas with organic Solvents as well as centrifugation and filtration obtained placenta extracts initially for the purpose of standardization Certain post-treatment must be given to either an injectable placenta extract for pharmaceutical use Purposes that are sterile and protein-free, or one that can be used as a mixed ingredient for cosmetic purposes Extract that is sterile and contains protein or protein

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poor and almost odorless. The standardized placenta extract is then combined with suitable solutions for the purpose of stabilization, many of which apart from buffer substances of the low molecular weight components detected in placenta extract, and some of these in their concentration strengthen, and also other active components, such as orotic acid, oligopeptides, deoxypentoses, purine / Pyrimidine bases and derivatives, etc. The solutions also contain preservatives, which according to Art and amount can be adapted to the respective purpose.

The placenta extract produced in this way can be kept for years and shows considerably better and longer lasting properties Effects than the previously known water-soluble placenta extracts. After sterile filtration in ampoules Bottled extracts can be injected directly or directly without further preparation for medical and pharmaceutical purposes can be used as a mixed ingredient in ointments or creams in cosmetics. In the latter case, the standardizing Post-treatment can also be significantly simplified, as the water-soluble placenta extract does not completely de-proteinize, it is only treated with activated charcoal to lighten and remove some components.

The invention is explained in more detail by the following examples, examples 3 and k showing the simplified production method for standardized and stabilized placenta extract, which, however, are mainly used for cosmetic purposes, since a certain amount of enzymes is desirable and advantageous here .

Example 1;

The usual way in the cold by treating crushed piacents with ether by centrifugation Placenta extract prepared and filtered is used for standardization by layers of carbon filtered and by extraction with organic solvents, such as ether, of hormones and

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remaining lipoid substances freed. It will be like this proceeded that the aqueous extract is divided into portions of 20 1 after filtering, which are then each etherified with 1-5 liters of ether. The separation of the Main amount of ether, and the remaining aqueous phase is adjusted to a pH value after the addition of chloretone from 7 »5 - 9 and freed from the remaining ether by introducing air. After the ether is blown out, the aqueous extract is treated with an organic acid, e.g. ironic acid, malic acid, Tartaric acid and the like, adjusted to a pH value of 2-5 and filtered after adding activated charcoal and then mixed with air or oxygen to remove the last traces of protein bodies and if necessary filtered again through layers of carbon. The extract is then given a certain amount Preservatives added, preferably chloretone, and a pH value of 6.7 - 7 »! set. The amount of preservative depends on the intended use, with pharmaceutical Purposes a higher chloretone content and for cosmetic purposes a lower content is desirable.

The standardized extract is then used for stabilization in a ratio of 1: 3 or 1: 5 a solution, which is composed of a basic solution and the partial solutions A, B, C. The following recipe applies to 100 1 final solution, which is made up as follows:

50 1 partial solution A, 2.5 1 partial solution B, 15 1 partial solution C and 32.5 1 basic solution.

The basic solution contains: 1-3 % ethyl alcohol, 0, ΙΟ, 8 % chloretone and citrate buffer with a pH value of 6.7 (e.g. 10 g citric acid and approx. 150 ml In-NaOH to 1 l).

The partial solutions are composed as follows:

Partial solution A: 3 - 100 g thymine

10 - 150 g succinic acid

3 - 100 g biotin

5 - 20 g of cysteine

3 30 g p-aminobenzoic acid

10 - 300 g malic acid

k0 - I50 g # C-Ke to glut a acid

0.25 - 100 g cytidine monophosphate di-

10 - 150 g vitamin C sodium salt

1 15 kg table salt

0.1-1 kg of glucose

0.5 - 25 g potassium chloride

100 - 500 g urea

10 - 500 g methyl glucamine

(the components are in 50 1 basic solution with a pH value of 6.7 solved)

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Partial solution B: 10 - 100 ng B

10 - 125 mg B.

- - ** ■ ™ ο - ο

10 - 200 mg Bf

0.1 - 1 mg B °

10 - 150 mg inositol

10-10 mg potassium pantothenate

15 - 150 mg folic acid

10 - 250 mg nicotinic acid amide

(the components are dissolved in 2.5 1 basic solution with a pH value of 6.7)

Partial solution C: 1 - 100 g each of lysine, tryptophan, aspartic acid, glycine, methionine, eC - alanine, serine, threonine, phenylalanine, isoleucine, leucine, proline, hydroxyproline, tyrosine, magnesium chloride, glutathione, oligopeptides, purine / pyrimidine Bases, guanosine, cytonine and orotic acid

(The components - are in I5 1 basic solution with a pH value of 6.7 solved)

After the 3 partial solutions have been mixed and made up to 100 l with the basic solution, the filter is sterile and pyrogen-free is determined and mixed with placenta extract in an appropriate ratio. After checking the pH value and renewed sterile filtration the extract is filled into ampoules.

The bottled extracts can be kept for years and show Standard values in the range of e.g.

pH 6.5-7.4

Dry residue

Lyophilisate 7.5 mg / ml or ^P 2 ° 5 ^ '^ mg / ml

Nitrogen 0.03 %

Ninhydrin test positive

Millon test positive

Sterility aseptic

Chlorine clay up to 0.6 % UV curve (1: 1.1: 10 max. 262-267 or 1:20)

Example 2t

The placenta extract standardized as in example (1) is used for stabilization in a ratio of 1: 1 - 1: 5 with a subsequent sterile filtered Mixed solution.

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For example, the sterile filtered solution contains the following components per 100:

O, 6 - 2, 5 kg Glycine 1. 5 - 5, 5 kg NaCl 1 - 15th G Tryptophan 200 - 800 G Chlorotone 1 - 3 kg Ethyl alcohol 3 - 25th G Thymine 5 - 500 G MgCl ₀

(Also include a sufficient amount of citrate buffer a pH value of 6.7 - 7 »l)

After sterile filtration, the extract is filled into ampoules and can be injected at any time without further processing.

The buffer solutions of Examples 1 and 2 usually contain a litrate or phosphate buffer with a pH of 6.6-7 * 1 * The preservative used is preferably chloretone, which is partly mixed with nipagin and / or ethyl alcohol or benzyl alcohol . The preservatives are used in concentrations of 0.1-0.7 % and the alcohols in concentrations of 0.5 % .

example 3:

Contrary to Examples 1 and 2, the freshly prepared placenta B extract is not completely de-ionized, but after filtration through a salt filter for the purpose of standardization only filtered after adding activated charcoal and / or through layers of charcoal and after the addition of a preservative, e.g. chlorite clay, freed of the remaining ether from the production by passing air through it.

The standardized extract is then used to stabilize it in a ratio of 1: 1 or 1: 3 with an aqueous chloro-clay solution of such a concentration mixed so that a final chloretone content of preferably 0.2 Ji is achieved. The aqueous extract is then adjusted to a pH of approx. 7 and immediately filtered sterile through a Millipore filter. If necessary, suitable buffer substances can be added to the standardized and stabilized extract be added.

Example 4: The extract is standardized as in

Example ₅ 3 _09809/0933

To stabilize 1 part of the extract, which contains 0.2% chlorine clay, 3 parts of a solution with a pH value of 7 is added, which contains the following components:

0.2 % chloretone

2.0 % ethanol under 1.0 % glycine under 0.2 % amino acids under 0.1 % thymine

as well as suitable buffer substances.

After sterile filtration, the extract can be added in large quantities of 5 - 10 % instead of the usual water to ointments and creams.

The key figures of the Piacenta extracts prepared according to Examples 3 and k are, for example

Dry residue nitrogen Pho sphat a sen

Chlorotone

Thioglycolate test Amino acids

7.0

over 7 mg / ml over 0.025 % over 50 KAE, over 1.3 KME

0.2 %

germ-free (? days, 31)

QIy, Ala, VaI, Tyr, Ser, Phe, Try, Asp, GIu, Len, i-Len, Pro,

Thr et al.

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Claims (4)

PATENT CLAIMS fT ^ Process for the production of standardized, stabilized, water-soluble placenta extracts, characterized in that the in a known manner by treatment with organic Solvents and centrifugation obtained placenta extract for the purpose of standardization initially by adding more organic Solvents freed from hormones and residual lipoid substances and from the main component of organic solvents is separated again and after removing the remaining organic solvents with the addition of suitable preservatives at a pH greater than 7, de-proteinized by acidification with organic acids and reduced to a pH of 6.5 - 7 »^ is set and then the standardized extract is mixed with a solution for the purpose of stabilization, which in addition to the usual buffer substances and preservatives contain a number of the low molecular weight components detected in placentas. 2. The method according to claim 1, characterized in that the residues of organic solvents are removed by blowing with air from the aqueous phase of the placenta extract in the presence of chloretone as a preservative and acidifying with citric acid, malic acid or tartaric acid at a pH value from 2-5 takes place, and that the extract is then filtered through layers of carbon in the presence of activated charcoal and, after oxygen has been passed through, 0.2-0.8% chloretone is added. 3. The method according to claim 1 or 2, characterized in that 1 part of the standardized extract is mixed with 3-5 parts of the solution, which is made up of a basic solution consisting of 1-3 % ethyl alcohol, 0.1-0.8 % Chloretone and citrate buffer with a pH of 6.7 and partial solutions A, B, C composed in which the following 509809/0933 Components in weight ratios based on 10 1 of the basic solution are: Partial solution A: Partial solution Bi 0.6 - 20 thymine 2.0 - 30 succinic acid 0.6 - 20 biotin 1.0 - 4 cysteine 0.6 - 6 p-aminobenzoic acid 2.0 - I50 malic acid 8.0 - 30 oC -ketoglutic acid 0.05 - 20 cytidine monophosphate disodium 2.0 - 30 vitamin C salt 00.0 - 3OOO table salt 20.0 - 200 glucose 0.1 - 5 potassium chloride 20.0 - 100 urea 2.0 - 100 methylglucamine 0.04 - 0.6 B o, o4 - 0.5 b; o, o4 - 0.8 B *
- 0.004 B ° 0.0004 - 0.6 inositol o, o4 - 0.6 ealium pantothenate o, o4 - 0.6 polyacid o, o6 - 1.0 nicotinic acid amide 0.04 Partial solution C: 3 »5 - 60.5 each of lysine, tryptophan, aspartic acid, glycine, methionine, oC -alanine, serine, threonine, phenylalanine, isoleucine, leucine, proline, hydroxyproline, tyrosine, magnesium chloride, glutathione, oligopeptides, purine / pyrimidine -Bases, guanosine, cytonine, orotic acid 4. The method according to claim 3, characterized in that the basic solution is mixed with the partial solutions A ₁ B ₁ C in a ratio of 32.5 i 50: 2.5: 15. , The method according to claim 1 or 2, characterized in that 1 part of the standardized extract with 1-3 Parts of the solution is mixed, which contains the following ingredients per 100 l: 0.6-2.5 kg of glycine 1.5 -. 5.5 kg NaCl 1.0-15.0 g tryptophan 800.0 g chloretone 3.0 kg of ethyl alcohol 25, og thymine 500.0 g of MgCl 200.0 1.0 3.0 5.0 509809/0933