Material with biological activity, method of its obtainment, and therapeutic medium based on it

FIELD: medicine.

SUBSTANCE: invention concerns medicine and claims therapeutic material and therapeutic media based on it for purulo-necrotic wound treatment, as well as method of obtaining therapeutic material. Material includes activated textile carrier with trypsin, insulin and lysocim immobilised in it.

EFFECT: significant reduction of wound surface cleaning time, accelerated healing.

17 cl, 2 tbl, 9 ex

 

The technical field to which the invention relates.

The present invention relates to medicine, specifically to the material (system)having biological activity, the method of its production and therapeutic tool based on the specified material (system) in the form of wound coverings which are intended for the treatment of purulent-necrotic wounds, trophic ulcers and other diseases.

The level of technology

Currently known materials and methods for their production on the basis of textile media from valdecaballeros (DAC) or polycaproamide containing immobilized on them drugs protein nature, which are successfully used in the treatment of purulent-necrotic wounds, trophic ulcers and other diseases.

So a method of obtaining material (DE 19747832 A1) in the form of wound dressings for use in the treatment of purulent-necrotic wounds and other injuries. The method consists in the fact that the cotton fabric in the form of medical gauze pre-oxidized solution periodate sodium to DAC with the content of aldehyde groups 0,04-0,06 mg-EQ. on 1 g of the carrier. The material is then treated with a mixture of solutions of trypsin and insulin in module 2,5 baths (module baths - weight ratio of the solution and the weight of the processed material). Thus proishodiashie aldehyde groups of the material of the carrier with the free amino groups of trypsin and insulin with the formation of the azomethine bonds, and, as a result, going immobilization of proteins called on the material. In the reference it is reported that when used as a carrier polycaproamide knitted fabric aldehyde groups are introduced by treating the polycaproamide knitted fabric with glutaraldehyde. The final product contains as an active ingredient trypsin, trypsin and insulin. As the advantages of this technical solution is called significant (20%) reduction of term of purification from the detritus of purulent-necrotic wounds compared to the closest analogues, which are named patents EN 2062113 and EN 2131268.

Also known is a method of obtaining material (US 6500799 B2) in the form of wound dressings, which involves a preliminary activation of the material of the carrier as a result of its oxidation, resulting in material contains 0,026-0.06 mg - EQ. aldehyde groups per 1 g of the carrier. Followed by immobilization on it, at least one enzyme in an effective amount for the functioning of the material within at least 3 days. This reference discloses a material with the above properties, obtained an open way. The material contains as the active ingredient trypsin, trypsin and insulin, trypsin and lysozyme. Here it is reported that the terms of cleansing wounds are 2, 4 days compared with 7, 5 days, during which achieves a similar effect by using conventional methods and means.

In the patent RU 2142818 disclosed is a method of obtaining dressings, including processing of absorbent textile material of the carrier with a solution of the protein preparation with the subsequent curing of the treated material of the carrier air and drying. In the way as absorbent textile material use dialdehydes or polycaproamide with the content of aldehyde groups 0,04-0,06 mg - EQ. on 1 g of the carrier, and the treatment is carried out by add absorbent textile material in a solution of proteins. The material contains as immobilized proteins trypsin or trypsin and insulin or trypsin and lysozyme. In the reference it is reported that the use of the material obtained by the disclosed method allows you to clean the wound within the range of 2.2-2.6 days.

However, despite the availability of medical practitioners quite successful for the treatment of purulent-necrotic wounds, trophic ulcers and other diseases, there is an urgent need for new, more efficient materials and tools.

The invention

The present invention provides a new therapeutic material for wound closure, possessing biological activity, is able to expedite the shape cleansing and healing of wounds.

Thus, the first aspect of the present invention is a therapeutic material based on textile media, which used cotton fabric from valdecaballeros (DAC) or polycaproamide knitted fabric. This therapeutic material is intended for the treatment of purulent-necrotic wounds of different origin.

Such wounds, in particular, can be trophic wounds in patients with diabetes mellitus.

The distinguishing feature of the claimed material is the fact that unlike known in the prior art it contains as active ingredients three immobilized protein, one of which is a hormone.

Such proteins are proteolytic enzymes (trypsin, chymotrypsin, turrilites and others), bacteriolytic enzymes (lysozyme, isoamylase) and hormones (insulin from the pancreas of man, ox, pig, recombinant insulin).

None of the known to the applicant of the information sources was not offered and is not intended for joint use of the named active ingredients in therapeutic material. Sharing how unexpectedly it was found by the authors of the present invention, these proteins can significantly speed up the cleansing and healing of purulent-necrotic wounds.

T is Ipsen, lysozyme and insulin are present in the weight ratio of 1:1,33:1,67, respectively, per 1 g of the carrier.

Therapeutic material according to this aspect of the invention is characterized by the fact that the content of aldehyde groups in the activated material of the carrier is not less than 0.02, for example 0,08-0,10 mg-EQ. on 1 g of the carrier, substantially higher than the number of active groups required for complete binding proteins and ensures their immobilization in the ratio in which they appear in the original solution.

Therapeutic material on the basis of polycaproamide knitted fabric or dialdehydes in accordance with the present invention is an application that is used in the treatment of extensive, shallow wounds.

therapeutic material based on valdecaballeros in accordance with this aspect of the invention may be subjected to mechanical degradation on the available equipment to the state of lint (cotton). Lint is more beneficial and convenient application in the treatment of narrow and deep pathological lesions (bullet, mine splinter and puncture wounds, periodontal pockets, ulcerative niches).

Further therapeutic material based on dialdehydes can be crushed to a fine powder with particle sizes of 0.1-0.6 mm powder with particle sizes of 0.5-0.6, the m can be introduced into the ear canal using insufflator for treatment of external otitis.

The same powder with particle sizes of 0.3-0.4 mm can be incorporated into the gels, ointments, suppositories, etc. on a hydrophilic base.

The same powder with particle sizes of 0.1 to 0.2 mm can be introduced into the volume of the aerosol container, containing components that form when they are applied on the surface of the wound foam. In the manufacture of aerosol cans and other dosage forms can be used technology firms "Altayvitaminy" (Russia).

So the next aspect of the invention is a therapeutic tool that is made on the basis of therapeutic material according to the first aspect of the present invention.

In one embodiment of the invention in accordance with this aspect of the invention a therapeutic tool in accordance with the present invention contains in addition to therapeutic layer, which represents a therapeutic material based on textile media - cotton fabric from dialdehydes or polycaproamide knitted fabric in accordance with the first aspect of the invention, and optionally other layers, one of which is a drainage layer, absorbing wound exudate. Typically, this hydrophilic material. You can use an extra outer layer which protects the wound from dirt, dust, pathogens and dry.

Su the layers together in the dressing sewing method.

In accordance with a third aspect of the present invention relates to a method of producing a therapeutic material in accordance with the first aspect of the invention.

The method includes the following stages:

activation of textile medium to the content of aldehyde groups 0,08-0,10 mg - EQ. per gram of the carrier;

- immobilization on activated carrier three biologically active components of a protein nature, taken in an effective amount for the rapid purification and healing of purulent-necrotic wounds.

In the method according to the invention as textile media use cotton fabric of cellulose or knitted fabric of polycaproamide. The activation is carried out with known methods, for example, by processing of cotton cellulose by periodate sodium to dialdehydes or by processing polycaproamide canvas with glutaraldehyde.

In the method according to the invention is used for immobilization of trypsin, lysozyme and insulin. The number immobilizovannykh molecules are in a weight ratio of 1:1.33:1,67 respectively.

Detailed description of the invention

Further, the invention is illustrated by examples of its implementation and comparative examples to illustrate the advantages.

Example 1

therapeutic material on the basis of the dialdehyde is ellulose

In the first reactor, containing 3,3 liters of distilled water, dissolve 12 g of iodic acid. In the second reactor with the same amount of distilled water was dissolved 3.8 g of sodium hydroxide. After complete dissolution of these substances are the alkali solution slowly poured the acid solution with vigorous stirring.

In the resulting solution at room temperature was placed 1 kg of medical gauze and incubated in the dark for 14 hours.

The obtained activated carrier containing valdecaballeros (DAN), drained, washed 4 times in 10 l of water and dried in air. Get DAC with the content of aldehyde groups of 0.08 mEq. on 1 g of the carrier.

Prepare a solution containing 0.55 g of lysozyme and 6.6 liters of water. Put it on 2 hours previously obtained DAC, then DAC wring out and put on 2 hours 6.6 liters of water containing of 0.44 g of insulin. Again press DAZ and placed for 2 hours 6.6 liters of water containing 0.33 g of trypsin. DAC again squeezed and dried in air to a residual moisture content of not more than 10%. Residual humidity is measured by the gravimetric method. The material dried at 105° C to constant weight, the weight loss is expressed in percentage. The material is cut into blanks of the desired size, e.g. 40×40 cm, and put in a four-applique size of 10×10 cm

Example 2

Canvas carrier obtained in example 1, the stock is granted in the form of appliqués 10×10. Received applications combined with draining (absorbing) layer and covered with a protective layer, the layers are combined in a bandage sewing method.

Example 3

Canvas carrier obtained in example 1, mechanical ruin to the state of lint (cotton wool) and Packed in packages.

Example 4

Canvas carrier obtained in example 1, grind to a powder with particle sizes of 0.5-0.6 mm Powder packaged in vials with rubber stopper and aluminium cap.

Example 5

Canvas carrier obtained in example 1, grind to a powder with a particle size of 0.1 to 0.2 mm and introduced into the foaming mixture, which is charged aerosol containers.

Example 6

Canvas carrier obtained in example 1, grind to a powder with a particle size of 0.3-0.4 mm, introduced into the hydrophilic gel on the basis of and fill in tubes or formed using powder suppositories.

Example 7

therapeutic material on the basis of polycaproamide

Textile fabric on the basis of polycaproamide placed in 40 l of 3 M hydrochloric acid and incubated for 4 hours at 60°C. the Cloth is squeezed and washed with water until the absence of chloride ions in the wash water.

Gidrolizovannogo so the canvas was incubated for 4 hours at 50°C in 37.4 l of 10% glutaraldehyde. Get activated Tex is safe media which is washed with water until no odor of glutaraldehyde. The carrier is dried in air to a residual moisture content of 10%. The resulting carrier contains 0.08 mg-EQ. aldehyde groups per 1 g of the carrier. Next, the media consistently treated with solutions of lysozyme, insulin and trypsin according to the procedure described in example 1.

Example 8

Canvas carrier obtained in example 7, is cut on the application of the desired size, for example, 7,5×10, 10×10, 10×15, 15×20, 20×30 cm, Packed in plastic bags.

Example 9

Packed products in the form of appliqués, lint or powder is subjected to gamma irradiation at a dose of 25 kGy.

Comparative example

Table 1 shows data demonstrating the effectiveness of therapeutic materials disclosed in patents and applications EN 2062113, EN 2131268, EN 2142818, DE 19747832 A1 and US 6500799 B2, and also demonstrated the claimed therapeutic material.

Table 1
The term cleansing of necrotic wounds in the treatment of various materials
Therapeutic materialThe time of the cleansing of wounds (day)
Patent RU 2062115, 21312682,9-3,1
By DE 1974832 A1 and EN 2142818 2,2-2,6
According to US 6500799 B22,4
According to the present invention1,8

In a separate study was conducted to study the proteolytic activity of a number of samples of therapeutic materials. Investigated (1) therapeutic material based on DAC, which were immobilized: lysozyme, insulin and trypsin, (2) therapeutic material based on DAC, which were immobilized insulin and trypsin, and (3) therapeutic material based on DAC, which was immobilized trypsin. It turned out that the proteolytic activity of therapeutic material (1) is 20% higher than that of the material (2) and 30% higher than that of the material (3). Proteolytic activity was determined by well-known methods: the Standard method for the determination of proteolytic activity for complex products proteases. /Kaverzneva DU // Applied biochemistry and Microbiology. 1971. V.7. No. 2. S-229.

An important characteristic of therapeutic materials for wound closure, is atraumatic. Removing the coating from the surface of the wound should not be accompanied by the injured, secondary bleeding and should be painless for the patient. Atraumatic can be estimated through stress the separation of the coating from the surface of the wound, which can be measured using a dynamometer as described in the article: a Method for comparative evaluation of possible traumas dressings. /Bychikhin NP and other // Surgery. 1986. No. 66. S.112-114.

It turned out that the breakout of material made according to the invention, 4 times lower than that of medical gauze and 2 times than that of the material obtained by DE 19747832 A1, EN 2142818 (see table 2).

Table 2
Comparative atraumatic different materials change over 2 days
Material typeBreakout force, g
Medical gauze398
DEC-trypsin-insulin203
DEC-lysozyme, trypsin202
DEC-lysozyme-insulin-trypsin99

The mentioned advantage can be a result of higher humidity claimed therapeutic material. After air drying residual moisture of medical gauze is about 4%, of the material according to DE 19747832 A1 - about 6%, and therapeutic material according to the invention is about 8%.

To increase the Oia number of crosslinks between the molecules of oxidized cellulose carrier and immobilized on the protein molecules materials and products in the form of applications, lint or powder can be subjected to gamma irradiation at a dose of 25 kGy. Such processing on the one hand allows the formation of additional crosslinking with the medium, but on the other hand, is for sterilizing material.

Therapeutic materials containing three protein immobilized on polycaproamide the media or on the basis of DAC preferably used in the form of applications in the treatment of extensive, shallow wounds.

If necessary, can be made dressings, containing, along with therapeutic drainage, water absorbing and retaining moisture protective layers. The mechanical strength of therapeutic material based DAC and therapeutic tools based on it is significantly lower than that of non-modified Marley. For this reason, therapeutic materials based on DAC if necessary, can be crushed state of lint (cotton) or powder.

Advantages of the claimed invention

Therapeutic materials according to the claimed invention significantly reduce the time of healing of purulent-necrotic wounds, and thus accelerate their healing. Therapeutic materials according to the invention differs substantially less traumatic, i.e. when removing material from the wound surface requires less effort, thus avoiding additional injury, eliminate krovat the increase, the usual other similar materials, and thereby alleviate the patient's condition.

It is known that lysozyme, insulin and other proteins cannot coexist in the same solution with trypsin, as they are the substrate for this proteolytic. However, if compabilitiy on the same media they are unexpectedly high joint therapeutic effect. And finally, if communicatie these three proteins they are not previously known high antioxidant activity, which may, and determines the above-described advantage is the shorter period of cleansing and healing of wounds.

1. Therapeutic material for the treatment of purulent-necrotic wounds containing activated textile carrier with the number of aldehyde groups is not lower than 0.02, for example 0,08-0,10 mg - EQ. 1 gram of the carrier, and immobilized proteolytic and bacteriolytic enzymes, and hormone protein nature.

2. The material according to claim 1, in which the indicated immobilized proteins are trypsin, a proteolytic enzyme, lysozyme - bacteriolytic enzyme and insulin - the hormone.

3. The material according to claim 1, in which trypsin, lysozyme and insulin are present in a weight ratio of 1:1,33:1,67 per 1 g of the carrier.

4. The material according to claim 1 on the basis of polycaproamide knitted fabric, which is an application.

5. The material according to claim 1 on the basis of the dia is DigitalGlobe, which is made in the form of application.

6. The material according to claim 1 on the basis of valdecaballeros who is subjected to mechanical degradation to the state of lint (cotton).

7. The material according to claim 1 on the basis of dialdehydes, which is crushed to a fine powder with particle sizes of 0.1-0.6 mm

8. Therapeutic tool for the treatment of purulent-necrotic wounds, representing the aerosol container, containing components that form when they are applied on the wound surface of the foam, and optionally containing a powder of a material according to claim 7 with sizes of 0.1-0.2 mm

9. Therapeutic tool for the treatment of purulent-necrotic wounds, representing a gel, ointment or suppository on a hydrophilic base containing powder according to claim 7 with a particle size of 0.3-0.4 mm

10. Therapeutic tool for the treatment of purulent-necrotic wounds containing therapeutic layer, which represents a therapeutic material based on textile media - cotton fabric from dialdehydes or polycaproamide knitted fabric as described in any one of claims 1 to 5, absorbent and protective layers, which are interconnected sewing method.

11. A method of obtaining a therapeutic material for the treatment of purulent-necrotic wounds according to claim 1, comprising the following stages:
activation textile media content Aldagi the groups not lower than 0.02, for example, 0,08-0,10 mg - EQ. per gram of the carrier;
immobilization on activated carrier three protein active components taken in an effective amount for the rapid purification and healing of purulent-necrotic wounds.

12. The method according to claim 11, in which the textile media use cotton fabric of cellulose or knitted fabric of polycaproamide.

13. The method according to claim 11, in which the activation is performed using the processing of cotton cellulose by periodate sodium to dialdehydes.

14. The method according to claim 11, in which the activation is performed using a processing polycaproamide canvas with glutaraldehyde.

15. The method according to claim 11, in which are used for immobilization of trypsin, lysozyme and insulin.

16. The method according to claim 11, in which lysozyme, insulin and trypsin immobilized consistently on the carrier in a weight ratio to 1.67:1,33:1, respectively.

17. The method according to claim 11, in which therapeutic material is subjected to gamma irradiation at a dose of 25 kGy.



 

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