RU2438683C2 - Method for making preparation of medicinal leech - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacological industry, particularly to a method for making a preparation of medicinal leech. The method for making the preparation of medicinal leech in which a leech is kept in an active fast regimen for at least 4 months, washed out, frozen and powdered to a homogeneous mass; the prepared mass is mixed with a water solution of polyvinyl alcohol and magnesium chloride in certain proportions and dried up in an environment avoiding any protein denaturation to elastic film formation.

EFFECT: method allows making the preparation of high anticoagulant and antichymotryptic activity and microbiological purity.

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Description

The invention relates to new drugs for medical purposes, namely to therapeutic agents based on biologically active substances produced by a medical leech, and a method for their preparation.

The specified drug refers to multicomponent physiologically active drugs that have anti-inflammatory, absorbable, analgesic effects, reduce the rheological properties of blood, inhibiting platelet and erythrocyte aggregation, is an immunomodulator, and exhibits neurotrophic properties.

There are various methods of isolating biologically active substances produced by a medical leech:

1. The method of obtaining the secret of the salivary glands of a medical leech. Baskova I.P., Russian Patent No. 2045954, 1994

2. Extracts from medical leeches:

a) extract from the head of the medical leech and torso:

Markward F. Untersuchungen uber Hirudin. Naturwissenschaften, 1955, 42, p. 537-538.

Markward F. Die isolierung und Chemische Charakterisierung des Hirudins. Hoppe Seylers Z. Physiol., 1957, 308, p. 147-156.

Markward F. Blutgerinnungshemender Wirkstoff aus Blutsaugenden Tieren. Iena, 1963.

Walsman P., Markward F. On the isolation of the thrombin inhibitor Hirudin. Thromb Res., 1995, 40, p. 563-570.

Baskova I.P. et al. Comparative characteristics of hirudin from whole leeches, heads and torsos. Biochemistry, 1980, 45, p. 266-272.

b) extracts from whole medical leeches:

Refers F.J., Zebe E. Protease inhibitors in the alimentary tract of the medicinal leech Hirudo medicinalis in vivo and in vitro studies. J. Comp. Physiol. Biochem., 1992, 162, p. 82-92.

Baskova I.P. et al. Comparative characteristics of hirudin from whole leeches, heads and torsos. Biochemistry, 1980, 45, p. 266-272.

Known therapeutic gel "Piyavit +", which is a water-salt extract of a whole medical leech (Gorban V.I. and other RF Patent No. 2268739 from 01/05/04).

Also known is the drug "Piyavit", which is a powder from lyophilized dried whole medical leeches (Baskova I.P. and others, RF patent No. 2033173 from 01.31.92, prototype).

However, in our opinion, lyophilization does not fully preserve the biological activity of substances of a protein nature, and extraction does not completely isolate biologically active substances from the body of a medical leech.

The aim of the invention is the development of a new drug for medical purposes, expanding the arsenal of funds for a similar purpose - a therapeutic agent based on biological active substances produced by a medical leech and method for its preparation.

This goal is achieved by eliminating freeze drying and extraction, but using a preservative and a binder - an aqueous solution of polyvinyl alcohol and magnesium chloride.

Example 1. Preparation of a preservative solution.

2.0 l of water, 0.5 kg of magnesium chloride and 0.25 kg of polyvinyl alcohol (PVA 20/1) (GOST 10779-78) are placed in a flask with a stirrer and reflux condenser.

The mass is heated in a boiling water bath and stirred until complete dissolution of the starting components. After self-cooling, a clear, viscous mass is obtained.

Example 2. Preparation of the drug.

Leeches, sustained in the regime of active starvation for at least 4 months until the intestinal canal is completely released from the blood, are introduced into suspended animation at a temperature of 5 ... 10 degrees. then washed with cold water (temperature 5 ... 10 degrees), placed in a linen bag and suspended in the refrigerator at a temperature of 5 ... 10 degrees for a period of 2 ... 4 hours to remove excess moisture. After that, the leech is transferred to a plastic bag and frozen at a temperature of -25 ... -28 degrees. The leech thus frozen is ground in a homogenizer. The resulting mass is mixed with a preservative solution in a ratio of 1: 1 and distributed on a stainless steel wall, which is placed in an oven, where it is dried at a temperature of 30 ... 37 degrees until a film is formed.

An elastic film of a dark brown color is obtained which exhibits self-adhesion. From the resulting material, tablets, dragees, films are made.

The resulting drug was tested for anticoagulant activity in comparison with the prototype.

Table 1 Anticoagulant activity of water extracts of the drug "Piyavit" (prototype) and the claimed object (average of 5 parallel) Sample number Structure Solution color Anticoagulant activity, battery one The drug "Piyavit" + saline (prototype) Dark brown 3600 +/- 100 2 The inventive object + saline Dark brown 3750 +/- 110

As can be seen from the data presented, the anticoagulant activity of the samples is almost the same. However, one should take into account the fact that these samples (prototype and the claimed object) are multicomponent preparations and not all sides of their biological action were studied quantitatively.

Also, studies were conducted on the microbiological purity of the claimed object (for compliance with FSP 42-0543-06 s12) after storage at room temperature for 6 months.

table 2 Microbiological studies * No. p / p Defined indicators, units measuring Research results Permissible Level one OMCH CFU / g 4.0 * 100 KOE / g No more than 1000 aerobic bacteria CFU / g 2 Yeast mold Not detected No more than 100 CFU / g 3 E.coli Not detected Lack of 1.0 g four S.aureus Not detected Lack of 1.0 g 5 Salmonella Not detected Lack of 1.0 g 6 P.aeruginosa Not detected Lack of 1.0 g 7 Other intestinal bacteria Not detected No more than 100 CFU / g * Normative documentation on research methods - Gosfarmakopeia XI edition - Volume 11 from Change 3. Category 4.2; issue 2, M .: Medicine, 1990. - p. 187.

Claims (1)

A method of obtaining a drug from a medical leech, in which the leech is kept in active fasting for at least 4 months, washed, frozen and crushed into a homogeneous mass, characterized in that the resulting mass is mixed with an aqueous solution of polyvinyl alcohol and magnesium chloride in a ratio of 1: 1 ( by weight) and dried under conditions in which protein denaturation does not occur until an elastic film is formed.