UA121937C2 - EARLY HEALING BANDAGE WITH PROTEOLYTIC ACTIVITY - Google Patents

Abstract

The invention relates to a wound-healing dressing with proteolytic activity, comprising a mixture for impregnation on a base containing polyvinyl alcohol, chitosan, protease from Bacillus thuringiensis var. israelensis IMB B-7465, glycerin, acetic acid and water, according to the invention, the mixture further comprises chlorhexidine bigluconate, and as a basis using medical gauze, at a ratio of the mixture for impregnation: medical gauze equal to 1: (0, 04-0, 06 ).

Description

The invention belongs to biotechnology, namely to a wound-healing enzyme agent that can be used in medicine and veterinary medicine in the treatment of wounds, skin damage, including thermal (burns, frostbite), trophic ulcers of various etiologies (diabetic, venous, post-traumatic, and others), and also in cosmetology.

It is known to be used as a basis for the immobilization of a complex of proteolytic enzymes of a medical non-woven bandage to obtain a promising wound covering (see patent VO Mo 2268751, publ. 27.01. 2006, Byul. Mo 3).

The closest to the claimed invention is a known wound-healing agent (see patent

of Ukraine Mo 131769 publ. 25.01.2019, Bul. Mo 2).

The well-known wound-healing agent contains polyvinyl alcohol, chitosan, acetic acid, glycerin, protease from Vasiyya5 Ipihipdiepvy mag. izgaeiiep5iz IMV B-7465 and water, with the following ratio of the specified components, wt. bo: polyvinyl alcohol 50.0-60.3 chitosan 61-82 acetic acid 4.7-5.5 glycerol 16.0-20.2 protease from Vasiyv5

Iipdiepov M.Sc. disappeared

IMV B-7465 0.15-0.9 water, the rest.

The well-known wound-healing agent is promising for accelerating the rejection of necrotic tissues, cleaning wounds of various genesis. It is able to protect the wound from the outside from infections.

But the well-known wound healing agent has the following disadvantages.

First, the polymer matrix has insufficient antimicrobial activity. For the prevention of purulent-inflammatory complications, it is advisable to use chlorhexidine bigluconate, an antiseptic with bactericidal and bacteriostatic action, under the influence of which the cytoplasmic membrane of bacteria is destroyed and their osmotic balance is disturbed, as a result of which the death of bacteria occurs (see Tpota5 Siinpeg, her am. Scapiadipe apa Oegimaimev5. - OPtap '

Epsusioredia oi Ipdyvila! Spetiviga 6th unit, 2006. - My. 17. - R.- 175-189).

Secondly, the matrix used is not plastic enough and not comfortable enough when used on the wound, and also does not adsorb wound exudate.

Thirdly, the prolongation of the effect of the known wound-healing agent has not been studied.

The composition of this wound healing agent was selected as a prototype.

The prototype and the claimed useful model have the following common features: - polyvinyl alcohol - chitosan

Zo - acetic acid - glycerol - protease from Vasiya5 (pPyhipdiepviv mag. izgaeiiepviz IMV B-7465 - water

The invention is based on the task of developing a wound-healing dressing with proteolytic activity, in which, by using non-woven gauze as a basis for impregnation with an enzyme immobilized in a polymer matrix (polyvinyl alcohol/chitosan) with

Vasiiv5 Shigipdiepviv M.Sc. izgayeepviz IMV B-7465, chlorhexidine bigluconate, as an antiseptic, provides the creation of a drug of prolonged proteolytic action with improved physico-chemical and biological characteristics for the treatment of wounds and burns.

The task is solved in a wound-healing dressing with proteolytic activity, which contains a mixture for impregnation on the base, while the mixture contains a polymer matrix (polyvinyl alcohol/chitosan), protease with Vasiyv5 Iyhipdiepvi5 mag. ivgaeiepvii IMV B-7465, plasticizer (glycerin), acetic acid and water because the mixture additionally contains chlorhexidine bigluconate, impregnated on the base, with the following ratio of components, wt. 95: polyvinyl alcohol 49.0-60.3 chitosan 5.9-82 chlorhexidine bigluconate (20 95 aqueous solution) 0.14-0.5 acetic acid 4.1-5.5 glycerol 15.0-20.2 protease from Vasiyv5

Iipdiepov M.Sc. izgaeIepviv 0.13-0.9

IMV B-7465, the rest is water, and medical gauze is used as a basis, with a ratio of mixture for impregnation: medical gauze equal to 1: (0.04-0.06).

What is new in the wound-healing bandage with proteolytic activity is that the mixture for impregnation on the base additionally contains chlorhexidine bigluconate, with the following ratio of components, wt. 9: medical gauze 4.0-6.0 polyvinyl alcohol 49.0-60.3 chitosan 5.9-82 chlorhexidine bigluconate (20 95 aqueous solution) 0.14-0.5 acetic acid 4.1-5.5 glycerol 15.0-20.2 protease with

Vasiyi5 Iipdiepvov mag. ivgaeYepviv IMV B- 7465 0.13-0.9 water, the rest, and medical gauze is used as a basis, with a ratio of impregnation mixture: medical gauze equal to 1: (0.04-0.06).

One of the functions of wound dressings is to protect the wound from the penetration of pathogenic microflora from the environment, since the development of the infectious process under the dressing leads to the spread of microflora in the tissues and inflammation.

For the prevention of purulent-inflammatory complications, it is advisable to add chlorhexidine bigluconate, an antiseptic with bactericidal and bacteriostatic action, to the polymer matrix to enhance the antibacterial effect of the drug.

As a proteolytic enzyme, the mixture with proteolytic activity contains a protease with complex proteolytic activity (caseinolytic, fibrinolytic, collagenolytic, elastase), the ratio of components is given above.

The cause-and-effect relationship between the totality of the declared features and the achieved technical result can be explained as follows.

The use of medical gauze as a basis for impregnation of an immobilized enzyme allows you to obtain a wound-healing bandage that has greater strength and plasticity compared to films, and is also capable of adsorbing wound exudate. These advantages, in combination with the possibility of producing gauze-based preparations of any shape and size, determine the prospects for the development of such preparations as medical means for the treatment of wounds and burns of various etiologies and locations. But the use of gauze as a basis for the creation of enzymatic preparations imposes certain restrictions on the amount of polymer solution with an immobilized enzyme fixed on this basis.

Impregnation of protease immobilized in a polymer matrix (PVC/chitosan) with the addition of chlorhexidine bigluconate, one- and two-layer gauze bandages (52:10 cm) was carried out. On

Fig. 1 shows a comparison of the proteolytic activity immobilized in a polymer matrix

From protease when it is fixed on bandages with different number of layers of gauze. The total proteolytic activity of bandages with different number of gauze layers is shown in Table 1.

Increasing the number of layers of gauze led to an increase in drying time from 18 to 26 hours. and, as a result, a slight decrease in the specific proteolytic activity of the drug (see Fig. 1).

Nevertheless, the use of two layers of gauze due to its greater adsorption capacity made it possible to apply a larger amount of the polymer mixture with the immobilized enzyme to the bandage and, accordingly, to increase the total proteolytic activity of the gauze by almost 1.7 times (see Table 1).

Table

Drying time General proteolytic

Bandage Bandage weight, g h "" activity (pOo" on " bandage) "PO - proteolytic unit

Immobilization had a positive effect on the overall proteolytic activity and stability of the two-layer gauze dressing - during long-term incubation compared to the free enzyme (see Fig. 2, which shows the preservation of the activity of the free and released protease from the gauze from the time of incubation in a buffer solution (pH 7 ,5; 137 C).

After 8 hours after incubation, the enzyme released from the bandage retains about 80 95 of the initial proteolytic activity, while only 30 95 is free, that is, the immobilized drug has a pronounced prolonged effect.

Wound healing bandage with proteolytic activity with immobilized protease

Vasiyy5 ihipdiepviv mag. izgaeiep5iz IMV B-7465 is promising for use in medical practice. In this regard, it is necessary to sterilize the received wound-healing dressing. The sterilizing filtration method was chosen for sterilization. The advantages of the method are: an effective sterilization procedure, mild conditions and speed of the sterilization process, the possibility of carrying out in laboratory aseptic conditions. The scheme of obtaining dressing material with protease Vasiyy5 Shihipdiepvi5 mag. The recovery of IMM B-7465 using sterilizing filtration is shown in Fig. 3.

Sterilizing filtration was carried out using GI S-5bugipde Tez filters (0.45

Mt). The prepared solution of polymers with the enzyme was passed through a filter, applied to sterile gauze bandages (5x10 cm), dried and packaged in aseptic conditions.

The use of sterilizing filtration of the polymer solution with protease had little effect on the activity of the drug. Checking the solution after filtration showed its microbiological purity.

A wound-healing bandage with proteolytic activity is prepared as follows: 6-10 cm 2095 aqueous solution of polyvinyl alcohol is mixed with 5.9-8.2 cm chitosan dissolved in 1.5-2 95 acetic acid, 0.35-0.45 cm3 of glycerin, and 0.05-0.F2 cm3 of a solution of chlorhexidine bigluconate (20 95 aqueous solution), mix. Then add 2.9-17.4 cm of enzyme solution (5.5-33.0 PO). The resulting mixture is stirred for 30 minutes, passed through a filter; subsequent application on a gauze bandage, drying at a temperature of 10-20 C in the air and packing in polyethylene film are carried out in aseptic conditions. Store at a temperature of 0-4 C.

Example 1

A wound-healing dressing with proteolytic activity was prepared as described above.

The components were taken, in this ratio, by mass. About: medical gauze 5.0 polyvinyl alcohol 55.1 chitosan 7.9 chlorhexidine bigluconate (20 95 aqueous solution) 0.5 acetic acid 4.5 glycerin 17.4 protease from Vasiinz IPyhipdiepviv mag. ivgaeYepviv IMV B- 7465 0.3 water 9.3.

Examples 2-4

Examples 2-4 explain the dependence of enzyme activity and bactericidal and bacteriostatic action on the ratio of the components of the drug.

Example 2

A wound-healing bandage was prepared, as indicated above. The components were taken, in this ratio, by mass. for: medical gauze 2.5 polyvinyl alcohol 55.1 chitosan 7.9 chlorhexidine bigluconate (20 95 aqueous solution) 0.5 acetic acid 4.5 glycerin 174 protease from Vasiinz IPyhipdiepviv mag. ivgaeYepviv IMV B- 7465 0.3 water 11.8.

The use of a smaller amount of medical gauze (1 layer) led to the application of a smaller amount of the polymer mixture with the enzyme to the bandage and, accordingly, to a decrease in the total proteolytic activity of the gauze by almost 1.7 times (see Table 1) due to its lower adsorption ability

Example C

A wound-healing bandage was prepared, as indicated above. The components were taken, in this ratio, by mass. for: medical gauze 7.5 polyvinyl alcohol 55.1 chitosan 7.9 chlorhexidine bigluconate (20 95 aqueous solution) 0.5 acetic acid 4.5 glycerin 174 protease from Vasi iv5 (Pygipdiepviv mag. ivgaeiepviz IMV B- 7465 0.3 water 6.8.

The use of a larger amount of medical gauze (3 layers) led to the application of a larger amount of polymer mixture to the bandage and, accordingly, to insufficient plasticity of the bandage and discomfort when using it on the wound.

Example 4

A wound-healing bandage was prepared, as indicated above. The components were taken, in this ratio, by mass. for: medical gauze 5.0 polyvinyl alcohol 55.1 chitosan 7.9 chlorhexidine bigluconate (20 95 aqueous solution) 0.14 acetic acid 4.5 glycerin 17.4 protease from Vasi

IShygipdiep5iv M.Sc. iztaieIepviz IMV B- 7465 0.3 water 9.7.

The use of a smaller amount of chlorhexidine bigluconate led to a weakening of the antibacterial effect of the drug.

The following essential features allow us to obtain the following technical result: to develop a wound-healing dressing by impregnating the gauze base with a wound-healing mixture of polyvinyl alcohol, chitosan, and protease from Vasiy5 IPyipdieep5vy mag. izgaeiiepvie IMV B-7465 and chlorhexidine bigluconate. The resulting wound-healing bandage has a high complex proteolytic activity together with an antimicrobial effect.

Claims (1)

FORMULA OF THE INVENTION A wound-healing dressing with proteolytic activity, which includes a mixture for impregnation on a base containing polyvinyl alcohol, chitosan, protease from Vasiya5 (Pygipdiepvi5 mag. izgaeIepviv IMV B-7465, glycerin, acetic acid and water, which differs in that the mixture additionally contains chlorhexidine bigluconate, with the following ratio of components, by weight: polyvinyl alcohol 49.0-60.3 chitosan 5.9-82 chlorhexidine bigluconate (20 95 0114-05 aqueous solution) and acetic acid 4.1-5, 5 glycerol 15.0-20.2 protease from Vasi iv5 (Pygipdiepviv mag. 0.13-0.9 ivgaeYiepviv IMV B-7465 water, the rest, and medical gauze is used as a basis, with a ratio of impregnation mixture: medical gauze equal to 1 :(0.04-0.06). NO - 50 and I: sho 40 5 in, m 20 5 - and M Milnyi bana aoaer o Lv layer Mreparat Fig. 1 a and GI V OA, ba in zv ZNISCH t; . but : in the tick «Ж and o same and in | As if Shan v 0 B I and Z 4 5 b 7 No Time. Goz. peni EE tv THEIR peche TV in stvnenya eeoeIMNevEya Fig. 2 2 be solution of CT, prepared on 5 UeniovVy quality of the Ministry of Internal Affairs, 205 aqueous solutions of PAS pi feya! Mixing the PVA solution with the HTZ solution, adding a plasticizer (hyperglycemic acid) and chlorohexide birlucionate (3095) to an aqueous solution. Adding an enzyme and thoroughly mixing the resulting amount. at room temperature, and under a pressure of 2 du SOR Ascentic pops on UMmation and Standardization of renohealing bandages for proteolytic activity Packing in hermetic polyethylene bags and fulaalALALAALADAAAAAKALKALA MA TSAAAAAAAALAAAALAAAAAAAKALKAMANN ALANA AAAAAKA VA p AAAAAAL HK AAAAK KK VAVNYA - Fig. 5