Biodegradable haemostatic therapeutic agent

FIELD: medicine.

SUBSTANCE: what is described is a biodegradable haemostatic therapeutic agent for control of bleeding, which provides co-immobilising ε-aminocapronic acid 50 mg, lysozyme 5 mg in distilled water 6.5 l for 3 hours at room temperature for dialdehyde cellulose 1 g at a degree of oxidation 12%. The material is pressed out and dried to residual moisture no more than 10% in the air in darkness. After having dried, the material is milled in a fine mill to particles having a size of 20 to 50 mcm. A rate of control of bleeding is 102 seconds. A time of total resorption is 10 days.

EFFECT: agent provides a high degree of hydrolytic destruction and a good haemostatic activity.

4 cl, 2 ex

 

The invention relates to a pharmacy, specifically to methods for hemostatic (hemostatic) drugs based on partially oxidized cellulose.

The above-mentioned drugs known. So, in the EPO application No. 1153620 described preventing bacterial infection in wounds hemostatic bandage from oxidized cellulose SURGICEL (firm "Johnson & Johnson"), containing from 1 to 10000 ppm of iron ions (3+), and from EPO application No. 0468114 known soluble hemostatic material in the form of webs of cellulose subjected to the oxidation of monochloracetic acid and sodium hypochlorite for introducing carboxyl groups COOH.

In the EPO application No. 0659440 describes a method for hemostatic based on oxidized cellulose containing 0.5-4.0% calcium and thrombin, fibrinogen and/or antifibrinolytic. The disadvantages of this method include the use of cellulose, mild oxidation of which with the transformation of CH2OH groups in the carboxyl (COOH), it is not possible to obtain a material that provides the possibility of a strong link with him ingredients: only the ions of Ca2+associated with COOH-groups of the ionic forces, while the remaining components are in the form of physical (mechanical) mixture. In addition, such materials are difficult to further grinding, for example, in Orasac (similar).

Known way to obtain a powdery material having hemostatic action, which comprises mixing in aqueous medium medium partially oxidized cellulose to valdecaballeros (DAC) with blood clotting factors, for example, thrombin and fibrinogen, to these substances additionally add gelatin, ε-aminocaproic acid and lysozyme, as well as partially oxidized cellulose use valdecaballeros in the form of a fabric having a degree of oxidation, i.e. the content of aldehyde groups, from 4 to 6%, with the following relations between the components:

- dialdehydes 1 g;

- fibrinogen 18/22 mg;

- gelatin 27/33 mg;

- ε-aminocaproic acid 45/55 mg;

- lysozyme 9,5/10,5 mg;

- thrombin 350 units;

- water and 6.5 ml

Components can withstand up to full covalent binding of drugs with dialdehydes with subsequent drying and grinding into powder. To prevent interaction of thrombin and fibrinogen the process of obtaining the drug lead in two stages. Separately prepare a solution of fibrinogen, ε-aminocaproic acid and half of the total amount of gelatin in half of the total amount of water and a solution of thrombin and lysozyme and the remaining amount of gelatin in the remaining amount of water in the obtained solutions as well who live within 3-4 hours by half the number of valdecaballeros, intermediates drained, dried in air and subjected to joint grinding. Time stop bleeding with this drug in the experiment was 110 sec, see, for example, patent RF №2235539, 2003(prototype).

The specified technical solution is not without flaws. First, the blood - thrombin and fibrinogen are very expensive and scarce. In addition, more patients in the world refuse to use drugs derived from animal organs and tissues or blood. Secondly, to avoid interaction in aqueous solution of thrombin and fibrinogen their immobilization are separated, then dried cloth and conduct joint grinding to powder. This adds cost and complexity to the process. The third drawback is the low degree of oxidation DAC, which leads to the slowing of the rate of hydrolytic degradation of the drug.

The present invention is to remedy these disadvantages and to obtain a powder of the biodegradable hemostatic drugs having a high degree of hydrolytic degradation (ability to complete desorption) and good hemostatic activity.

To this end substantially changed the composition of the medicinal components excluded expensive deficient blood and gelatin, significantly simplifies the technological process is to obtain medicines significantly increased oxidation DAC, which ultimately led to the accelerated hydrolytic degradation and, consequently, to reduce the time complete resorption while increasing hemostatic activity.

Additional studies have shown that increasing the degree of oxidation of valdecaballeros (DAC) and its protein derivative increases the rate of hydrolytic degradation. While proteins diffuse in the aquatic environment is not in the form of natural substances and in the form of conjugates with oligomers DAC, which makes them more resistant to inactivating factors (heat, pH of the medium, inhibitors) compared with native biologically active substances (BAS), i.e. show prolonged a specific action. The kinetics of hydrolytic degradation describes a semi-log anamorphosis and in logarithmic coordinates is a straight line, the tangent of the angle which quantitatively expresses the rate constant hydrolytic degradation as the reaction rate constant of the first order.

Knowing these constants, it is easy to calculate the half-life period (polarizable) drug by the formula

K=0,69/T/2,

where K is the rate constant of destruction,

T/2 - time half-life of the drug.

Experimental data of the dependence of the rate constants for the hydrolytic degradation of the oxidation DAC are shown in table 1.

Table 1
Oxidation DAC, %The rate constant hydrolytic distractie, min-1
4-618×10-5
6,521×10-5
1233×10-5
1839×10-5

On the other hand, when increasing the degree of oxidation DAC sharply decreases its mechanical strength. The influence of the degree of oxidation of cellulose (medical gauze) to DAC on the strength characteristics of the material are presented in table 2.

Table 2
The degree of oxidation,%Breaking load (on the base), DanThe preservation of the breaking load, %
-the 33.4100,0
3,022,065,2
5,021,68,020,561,4
to 12.018,356,6
15,016,249,0
26,045,045,0

From the data presented in tables 1 and 2, it follows that an increase in the oxidation rate of over 12% is irrational, because with only a slight increase in the rate constants of hydrolytic degradation, significant drop in mechanical strength DATS more than 50%. It should also be noted that when the grinding DAC high degrees of oxidation is formed dust fraction of the powder.

Comparative characteristics of the hemostatic properties of materials and their relationship to the degree of oxidation DAC, part time and full resorption are shown in table 3.

Table 3
Oxidation DAC,%The coagulation factors, mg per 1 g DACConstant speed hydrolytical some of distractieTime stop bleeding sec The full resorb-
tion, of the day
According to the patent of Russian Federation №2235539 4-6%, the placeholderFibrinogen 18-22 gelatin 27-23 aminocaproic acid 45-55 lysozyme 9,5-10,5 thrombin 350%18×10-5min-111020
On the proposed decision of 12%Aminocaproic acid is introduced in a quantity of 50 mg/g, lysozyme - 5 mg/g33×10-5min-110210

Thus, the proposed new biodegradative hemostatic drug in powder form with particle sizes from 20 to 50 μm has the following characteristics:

DAC with degrees of oxidation - 12% - 1.0 g;

ε-aminocaproic acid - 50,0±0.25 mg/g;

lysozyme - 5,0±0.25 mg/g

Medicinal composition, such composition provides a high speed stop bleeding, prolonged preservation of the sterility of the material and the minimum time to complete resorption.

Time stop bleeding - 102 sec.

The time of complete resorption to 10 days.

Therefore, the proposed solution meets all the requirements of the invention: novelty, usefulness and level of technology.

The novelty is ostoic, first, that for immobilization of hemostatic agents as media use DAC with the degree of oxidation of 12%. Secondly, as a specific hemostatic use ε-aminocaproic acid (without fibrinogen, thrombin and gelatin). Thirdly, the proposed composition is a fine powder with particle sizes from 20 to 50 μm. Fourthly, the time for complete resorption is 10 days.

The obvious usefulness: speed stop bleeding equal to 102 sec, which is less than, but comparable with the time stop bleeding prototype - 110 sec, but given that this is simple and few of the medicinal components of the composition, especially with the property biodegradation (resorption), the usefulness of this development is indisputable.

The level of technology

The drug is made Pharmacopeia and other documents required for inclusion in the Pharmacopoeia of the Russian Federation. No single product on the basis of DAC, previously permitted for medical use, has no such status.

The invention is illustrated in the examples.

Example 1

In the reactor fill in 6.5 liters of distilled water, include the stirrer and add 185 g periodate sodium. The solution is stirred until complete dissolution of the reagent. In the resulting solution of periodate sodium is amemait 1 kg of cellulose (medical cotton gauze) in the form of cloth and incubated at room temperature in the dark for 14 hours. After activation, wring out the cloth, washed 4 times in 10 l of distilled water, again squeezed and dried in air in the dark to a residual moisture content of not more than 10% at room temperature. Oxidation of cellulose to DAC is 12%.

Example 2

6.5 liters of distilled water dissolve 50 g of ε-aminocaproic acid, 5 g of lysozyme, the reaction mass is stirred, the pH of the solution to 5.5 to 6.0. The solution was placed 1 kg DAC with the degree of oxidation of 12%, incubated for 3 hours at room temperature in the dark, drained, dried in air in the dark to a residual moisture content of not more than 10% and ground in a mill, super thin grinding to a particle size of from 20 to 50 microns.

In experimental conditions in adult rabbits caused bleeding during intravenous injection of 500 IU of heparin and 250 fibrinolizina ED. Mechanically, the bleeding stopped after 20-30 minutes

Under hypentelium anesthesia superficial excision of the fragment liver size 1.4 cm2caused the bleeding, the wound was imposed biodegradative hemostatic agent, noting stop bleeding through 102 sec.

1. Biodegradative hemostatic drug in the form of microfiber, consisting of dialdehydes with chemically immobilized on the coagulation factor ε-aminocaproic KIS the Auteuil and bacteriolytic enzyme lysozyme, characterized in that, as a partially oxidized cellulose it contains dialdehydes with the degree of oxidation of 12%, with the following components:
DAC with oxidation States 12% - 1.0 g;
ε-aminocaproic acid 50,0±0.25 mg/g;
lysozyme 5,0±0.25 mg/g

2. Biodegradative hemostatic medicinal product according to claim 1, characterized in that the particle sizes of its microfiber forms of partially oxidized cellulose with immobilized her components ranged from 20 to 50 microns.

3. Biodegradative hemostatic medicinal product according to claim 1, characterized in that the time of the full hydrolytic destruction - complete biodegradation is not more than 10 days.

4. Biodegradative hemostatic medicinal product according to claim 1, characterized in that the stop time of bleeding, its use is not more than 102 seconds.



 

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