RU2268739C2 - Therapeutic gel based on bioactive blood-sucker producing substances and method for production thereof - Google Patents

Abstract

FIELD: medicine preparations based on bioactive substances producing by blood-sucker.

SUBSTANCE: claimed method includes application polyvinyl alcohol and magnesium chloride polyhydrate complex as extractant for bioactive substances producing by blood-sucker. Blood-sucker living persons are extracted without lyophilization or grinding.

EFFECT: new pharmaceutical with antiinflammation, anticoagulant and analgesic action.

5 ex, 2 tbl, 2 cl

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 isolation.

The specified gel "Piyavit +" refers to physiologically active drugs with anti-inflammatory, absorbable, analgesic effects, and contains in its composition an extract of biologically active substances produced by a medical leech, obtained by a new method.

Known drug based on the bischofite mineral (MgCl ₂ · 6H ₂ O), containing bromine, iodine, trace elements, which is effective in diseases of the musculoskeletal system, peripheral and central nervous system, as well as in sprains and muscles (NPP "Volgograd bischofite ", Volgograd).

Bischofite is also known as a gel (TU 9158-002-45053421-01, an analog of bisholin), which has anti-inflammatory, absorbable and analgesic effects. Gives a lasting therapeutic effect. Effective for diseases of the musculoskeletal system, sprain of muscles and ligaments, consequences of injuries. It is used in the form of massage (rubbing) 2-3 times a day. The course of treatment is 10-14 days. A break between courses of 3-4 weeks. Contraindications: individual intolerance, skin diseases in the area of the gel.

A large number of medicinal ointments, liniments and balms are known that are used to treat boils, mastitis, inflammatory diseases of the joints and muscles, injuries of soft tissues and joints, burns, frostbite, neuralgia, radiculitis, etc.

So, methyl salicylate (methyl ester of salicylic acid) is used as an anesthetic and anti-inflammatory agent.

In addition, there are dosage forms containing methyl salicylate: Sanitas liniment (balsam), Bom Bengue ointment, Naftalgin liniment, Kansin, complex methyl salicylate liniment, saliment. They are used externally for joint and muscle rheumatism, arthritis, exudative pleurisy (M.D. Mashkovsky, Medicines, Part 1, M., Medicine. 1978. - p. 180).

It is known to use birch tar in the form of 10 ... 30% ointments, liniments to improve blood supply to tissues, stimulate the regeneration of epithelial tissue, enhance wound healing processes (M.D. Mashkovsky, Medicines, Part 11. M., Medicine. 1978 . - p. 368).

Liniment Vishnevsky A.V. used in the treatment of wounds, ulcers, bedsores (ibid.).

Ichthyol has an anti-inflammatory, local anesthetic and antiseptic effect. Applied externally for dermatitis, burns, neuralgia, arthritis and other diseases (ibid.).

Liniment of naphthalan oil when applied to the skin has a softening, absorbable, disinfectant and some analgesic effect. Applied externally for various skin diseases, inflammatory diseases of the joints and muscles, with neuralgia, neuritis, radiculitis, burns, ulcers, etc. (ibid.).

Chlorophyllocarotin paste. Used in the treatment of burns, sluggishly healing wounds and ulcers (ibid.).

Sea buckthorn oil. Applied externally in the treatment of burns, pressure sores, radiation injuries of the skin (ibid.).

Vinylinum (Shostakovsky's balm) polyvinyl butyl ether

Used for boils, trophic ulcers, purulent wounds, mastitis, soft tissue injuries, burns, frostbite, promotes tissue regeneration and epithelization (M.D. Mashkovsky, Medicines, part 11, M., Medicine, 1978. - p. 371 )

Known therapeutic ointment "Piyavit" containing in its composition an aqueous extract of biologically active substances (BAS) produced by a medical leech. In this case, the extract is obtained from the powder of powdered lyophilized leeches (from the drug "Piyavit", which is approved for clinical use as a thrombolytic drug, pharmacopeia article of the enterprise NVF "Girud I.N." - FSP 42-0177-0543-00). In addition to the water extract "Piyavita" ointment contains polyethylene oxide 1500, polyethylene oxide 400, menthol, propylene glycol, nipagin, sodium chloride - the prototype.

Several methods for isolating biologically active substances produced by a medical leech (Hirudo medicinalis) are also known: extract from the head part of leeches, extract from the body of a ground leech, extract from the body of a ground lyophilized leech. The latter method has found the greatest distribution. On its basis, the therapeutic drug “Piyavit” is obtained (IP Baskova, RF Patent No. 2033173 of 01/31/92).

A technical solution is known in which a tool in the form of a cream from a medical leech is disclosed having the same purpose as the claimed tool, and the preparation method includes the step of obtaining an aqueous extract from a medical leech (RF patent No. 2153325 of July 27, 2000 - prototype) .

However, in our opinion, lyophilization is not the best way to process substances of protein nature, since it causes their denaturation and, as a result, irreversible changes in the spatial arrangement of the constituent fragments of the structure. This can be detrimental to substances with biological activity.

Thus, the search for alternative methods for isolating biologically active substances produced by a medical leech is of particular relevance.

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

This goal is achieved by using polyvinyl alcohol (PVA) complex and magnesium chloride polyhydrate as an extractant of biologically active leeches, and live specimens of medical leeches without lyophilization or grinding (homogenization) are extracted.

Example 1. Preparation of an aqueous solution of PVA - MgCl ₂ .

125 g of PVA, 250 g of MgCl ₂ hexahydrate and 700 ml of deionized water are placed in a vessel and kept under stirring in a boiling water bath until the PVA is completely dissolved. The resulting solution is cooled and used to prepare the Piyavit + gel. Storage of Piyavit + treatment gel is possible at room temperature, while storage of such preparations requires a temperature of no higher than 10 ° C.

Example 2. Preparation of extract "Piyavit +".

Live medical leeches with a total weight of 10.0 g, which corresponds to approximately 2.0 g of powder of crushed lyophilized leeches that have passed quarantine, are placed in a small amount (25.0 g) above the prepared solution and kept at room temperature for six days, stirring occasionally (30 ... 60 rpm at a temperature of 22 ... 24 ° C) with a glass rod. Outwardly, there are no changes except darkening of the solution. However, when examining the body of a leech, it is seen that a through oval hole appears in the tail part of some leeches. Over time, the hole increases in size. In individual leeches, the tail part is separated and almost completely dissolved (degraded). The head part remains firm and elastic. In this case, a leech through the oral cavity secretes a biologically active substance along with belching from the intestinal canal in response to irritation of the corresponding receptors of the head region with a concentrated solution of MgCl ₂ .

The obtained extract was tested for anticoagulant activity (AKE) according to the methodology of the pharmacopeia article of the enterprise of the Scientific Foundation “Girud I.N.” (FSP 42-0177-0543-00). The data are presented in table 1.

Table 1 The growth dynamics of the anticoagulant activity of the extract "Piyavit +" in a solution of PVA - MgCl ₂ (average of 5 parallel measurements) Sample number extraction time solution color anticoagulant activity, AKE one 1 day light yellow 699 +/- 62 2 2 days yellow 1166 +/- 121 3 3 days brown 3122 +/- 182 four 4 days dark brown 5250 +/- 193 5 5 days dark brown 9606 +/- 201 6 6 day dark brown 10500 +/- 194 7 7 days dark brown 18900 +/- 205 8 8 day dark brown 18613 +/- 203 9 9 days dark brown 18870 +/- 198

Additionally, we studied the combined effect of the extract "Piyavit +" obtained by water extraction of ground lyophilized leeches and a solution of polyvinyl alcohol and magnesium chloride on anticoagulant activity. The results are shown in table. 2.

table 2 Anticoagulant activity of the studied compounds (average of 5 parallel) Sample number Structure solution color Anticoagulant activity, AKE one Extract "Piyavit +" + saline dark brown 18845 +/- 200 2 Extract "Piyavit +" + PVA solution - MgCl ₂ dark brown 19750 +/- 201

As can be seen from the above data, with their combined action of antagonism is not observed. Separately, a solution of PVA - MgCl ₂ does not have anticoagulant activity.

Based on the obtained extract, the composition of the Piyavit + therapeutic gel was developed. The composition of the therapeutic ointment "Piyavit", produced by the "Hirud I.N."

The difference was that the base (polyethylene oxides) was replaced with an aqueous PVA - MgCl ₂ solution and preservative (nipagin) was excluded, since a sufficiently high concentration of MgCl ₂ in the gel composition ensures its reliable preservation.

Example 3. Preparation of the basis of the gel "Piyavit +".

In a 3-necked flask with a capacity of 150 ml, equipped with a stirrer, reflux condenser and thermometer, 86.0 g of a solution of PVA - MgCl ₂ , prepared as described above, is placed, heated in a water bath to a temperature of 40 ... 45 ° С and at moderate stirring 30 ... 60 rpm load 5 g of glycerol and 1.5 g of menthol. A clear, viscous liquid is obtained.

Example 4. The preparation of the gel "Piyavit +".

The base of the gel at a temperature of 30 ... 35 ° C and with stirring load the obtained extract in an amount of 8.0 g. Stirring is continued until the mass is completely homogenized. Then the mass is unloaded in a container.

Example 5. Testing the gel "Piyavit +".

The healing properties of the Piyavit + gel were studied in experiments on volunteers. Depending on the nature of the injury, the gel was applied either directly to the affected area with a thick layer (minor thermal burns, sunburn, frostbite, insect bites, bruising, etc.), or in the form of gauze dressings, napkins (inflammatory diseases of the joints and muscles, significant injuries to muscle tissues and joints, extensive burns, surgical sutures, etc.). If the skin is disturbed, it is not recommended to apply the gel to an open wound, since the composition includes salt (magnesium chloride) in a significant concentration, which causes an acute pain reaction. It is necessary to wait for some epithelization of the wound surface. It should be noted that in the treatment of extensive thermal burns there are practically no scars.

The results of the experiments showed the high efficiency of the healing properties of the Piyavit + gel.

Claims (2)

1. A therapeutic gel with anticoagulant, anti-inflammatory and analgesic activities, including an extract of biologically active leech substances, as well as polyhydric alcohol and menthol, characterized in that it contains an extract of biologically active leech substances obtained from live medical leeches by extracting them in an aqueous solution of polyvinyl vinyl alcohol and magnesium chloride for 6 days with periodic stirring 30 ÷ 60 rpm / min and a temperature of 22 ÷ 24 ° C, and as a polyhydric alcohol it contains glycerin and tively polyhydrate complex of polyvinyl alcohol and magnesium chloride at the following contents of ingredients, wt.%: Biologically active leech extract 7-9 Polyvinyl alcohol complex polyhydrate and magnesium chloride 83-88 Glycerol 4-6 Menthol 1-2 2. A method of obtaining a therapeutic gel having anticoagulant, anti-inflammatory and analgesic activities by aqueous extraction of a medical leech, characterized in that live medical leeches are extracted in an aqueous solution of polyvinyl alcohol and magnesium chloride for 6 days with periodic stirring at 30-60 rpm and temperature 22-24 ° C.