DD292382B5 - METHOD FOR PRODUCING A PARENTERALLICALLY APPLICABLE STORAGE-RESISTANT IMMUNOSTIMULATING PREPARATION - Google Patents

Abstract

The invention relates to a stable, liquid or lyophilised preparation for use as immunostimulator in human medicine and to a process for its preparation and to its use. The preparation is prepared at a pH between 6 and 8 by combining splenopentin or splenopentin analogues with a hydroxybenzoic ester and is distinguished by improved and prolonged stability.

Description

2. The method according to claim 1, characterized in that the peptide is used in a concentration of 1 to 100 mg / ml and the p-hydroxybenzoic acid ester in a concentration of 0.1 to 10 mg / ml.

3. The method according to claim 1 and 2, characterized in that the mixing ratio of

Peptide solution and hydroxybenzoic acid ester solution 1: 1 to 10: 1, preferably 5: 1.

Field of application of the invention

The invention relates to a method for producing a storage-resistant splenopentin or splenopentin analogue-containing preparation suitable for parenteral administration for immunostimulation. It finds application in the pharmaceutical industry

 Indications of the drug are

- immunodeficiency in HIV / AIDS patients;

- restoration of the functioning of the immune system in cancer patients in chemo- and radiotherapy;

- immune paralysis in organ transplantation;

- restore the functioning of the immune system in bone marrow transplantation;

- rheumatoid arthritis;

- Psoriasis with strong joint involvement.

Characteristic of the known state of the art

Peptides are generally unstable, especially in aqueous solutions. The hydrolysis of the peptide bonds leads to cleavage of the molecule into ineffective peptide fragments. Higher temperatures lead, in particular in the presence of water, to denaturation and thus to the occurrence of precipitations which make parenteral administration impossible. Depending on the amino acid sequence of the peptide, extensive tests are necessary in each case in order to develop an individual optimal formulation which ensures sufficient stability of the preparation in the period of usefulness. Splenopentin and analogs are unstable, as are other peptido. According to monograph and pharmaceutical reports, diacetylsplenopentin hydrochloride is already prescribed a very cool, light and moisture protected storage for the substance. Experience has shown that the stability deteriorates in the presence of water or in Lyophilisiorung.

Storage-stable, parenterally administered preparations with immunostimulating action on the basis of splenopentin or its analogues have not hitherto been known.

Object of the invention

The aim of the invention is the preparation of a stable parenterally applied preparation with immunostimulating action based on splenopentin or splenopentin analogs.

Explanation of the essence of the invention

The invention is based on the technical object of providing a method for converting splonopentin or splenopentin analogs as immunostimulators into suitable parenterally administrable storage-stable solutions by suitable additives and conditions.

According to the invention, the Aufpah- "uurch solved that using splenopentin or Splenopentinanaloga while avoiding the Fällungsbereicr it by means of conventional pharmaceutically and pharmaceutically suitable buffer substances, a pH range between 6.0 and 8.0, preferably from 6.5 to 7, 5, is set. The peptide used is preferably diacetylsplenopentin hydrochloride in a concentration of 1 to 100 mg / ml. As stability promoters aqueous solutions of hydroxybenzoic acid esters in a mixing ratio of 1: 1 to 10: 1, preferably 5: 1, are used at 9O ⁰ C to 95 ⁰ C tempered aqueous solutions. The peptide is dissolved in 0.1 mol / l sodium hydroxide solution and then treated with the tempered to DO ⁰ C to 95 ⁰ C hydroxybenzoic acid solution. After adjusting the pH, the solution is sterile filtered through O, 2 pm membrane filter and filled under aseptic conditions in 2 ml ampoules. The hydroxybenzoic acid ester is preferably used in a concentration of 0.1 to 10 mg / ml

 As the organic solvent, propylene glycol and / or ethanol may be added to the solution.

After addition of 1% to 10% of a lyophilization vehicle. preferably mannitol or alanine, the solution can subsequently be added

Temperatures to 3O ⁰ C be lyophilized. The peptide preparations prepared by this procedure show good stability and thus also enable one

parenteral application (Table 1).

Hydroxybenzoic acid esters have hitherto been used in injection solutions only for the prevention of microbiological contamination (DD 273981), but not to improve the storage stability, in particular with regard to the prevention of precipitation, which make parenteral administration a lethal hazard and thus impossible.

The use of Hydroxybenzoesäureestern as a solubilizer and stabilizer for peptides is thus new, the success quite

surprised.

The solution according to the invention makes it possible to improve the stability of an aqueous solution and of a corresponding lyophilisate so that no restrictions of the storage regulations are more necessary and the resulting solutions can even be applied parenterally. The restriction of the storage conditions prescribed for the active ingredient (very cool storage and protection against moisture) is eliminated.

embodiments

example 1

In 600 ml of 0.02 mol / l phosphoric acid, 27 g of diacetylsplenopentin hydrochloride are dissolved. By a surplus of

Sodium hydroxide is first adjusted to a pH of 8.0 to 9.0. After stirring for 20 minutes, 0.02 mol / l phosphoric acid

adjusted to pH 7.5. In 300 ml of water for injection, 1.5 g of hydroxybenzoate are dissolved at 9O ⁰ C to 95 ⁰ C and combined with the mixture immediately and with stirring. There is a fine adjustment of the pH to 6.9-7.1. The mixture is then made up to 11, sterile-filtered over 0.2 μm membrane filter and packaged under aseptic conditions.

Example 2

In 500 ml of water for injection, 27 g of splenopentin hydrochloride are dissolved. The pH of the solution is first adjusted to 8.0 to 9.0 with meglumine and after 20 minutes of stirring with 1 mol / l hydrochloric acid to pH 7.5. In 200 ml of water for injection 0.75 g propyl hydroxybenzoate are dissolved at 9O ⁰ C to 95 ⁰ C and immediately mixed with stirring with the peptide solution. After the addition of 200 g of propylene glycol, the mixture is made up to 11, sterilized via O, 2 ^ m membrane filter and filled under aseptic conditions.

Example 3

20 g of diacetylsplenopentin hydrochloride are dissolved in 450 ml of 0.05 mol / l sodium hydroxide solution. Subsequently, 50 g alanine are registered.

In 450 ml of water for injection, 1.5 g of hydroxybenzoate are dissolved at 9O ⁰ C to 95 ⁰ C and immediately mixed with the peptide solution. After stirring for 20 minutes, the pH is adjusted to pH 7.5 with hydrochloric acid and the volume with

Water for injection to 11 supplements. The sterile solution, filtered through membrane filters, is injected under aseptic conditions

filled into injection bottles and lyophilized at temperatures up to 25 ° C.

Attachment 1

Ί Table 1: Comparative stability studies of splenopentin vials

storage Time PH value (comparison Solution without addition according to the invention) > 25 _M m Splenopentinqehalt In addition to stains Bedingun Clarity of particle content gene solution to USP 0 [%] 1%) > 10pm 0 output 0 7.0 0 t> 7.3 2.1 value 28 d 6.91 XXX 0 0 97.2 2.0 6 ⁰ C 28 d 6.95 XXX 0 _ 97.1 2.2 2O ⁰ C 28 d 6.89 XXX 0 - 97.0 2.5 4O ° C Uahr 6.68 XXXXX 0 - 94.2 4.0 6 ⁰ C Uahr - XXXXX - - - 2O ⁰ C Uahr - - - - - 40 ⁰ C - -

storage Time PH value Clarity of (Patent solution) Splenopen- Next Bedingun solution solution particle content tingehalt stains gene to USP (%] [%] > 10 / im> 25μιη

Output value 6 ⁰ C

20 ° C

4O ⁰ C

0 28 d

28 d

28 d

7,1 7,11

7.13

7.12

99.2 99.4

99.3 99.5

1.5 1.6 1.5 1.8

6 ⁰ C Uahr 7.13 0 && 99.1 1.8 2O ⁰ C Uahr 7.08 0 99.3 1.9 40 ⁰ C Uahr 7.11 0 = A 99.0 2.1

Explanations: O = no makrosk. recognizable particles xxx = many white particles

xxxxx = many white particles and agglomerations 0 = particle content does not comply with the requirements of USP XXI (max 10000 part./I> 10μηι max 1000 part./I> 25μπι)

Claims (1)

1. A process for the preparation of a parenterally administrable storage-stable splenopentin or a Splenopentinderivat, preferably Diacetylsplenopentin, containing preparation by dissolving the active ingredients or their hydrochlorides in a buffered to a pH between 6.0 and 8.0 buffered aqueous solution, characterized in that to this solution to 90 to 95 ⁰ C. heated aqueous p-hydroxybenzoic acid methyl and / or propyl ester solution added and the mixture further processed according to standard pharmaceutical technology