RU2585366C2 - Effective hemostatic agent based on double lithium-copper salt of polyacrylic acid, simultaneously having high antiseptic action - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to creation of new chemical compounds, which can be used in medical practice as a haemostatic agents of local action, for example when performing surgical operations, for stopping bleeding in traumatic wounds, in dentistry, simultaneously having bactericidal, fungicidal, bacteriostatic and funguistatic action. Described is a partial double lithium-copper salt of polyacrylic acid according to general formula: (-CH₂CHCOOH-)_n(-CH₂CHCOOLi-)_m(-CH₂CHCOOCu_0.5)_k, where n = 318871-956613; m = 96136-288408, k = 24573-73718, content of Li and Cu is 4.43-4.46 and 3.93-4.10 0.00034-0.00101 g-at and 0.00029-0.00093 g-at, respectively, and content of carboxyl groups is 0.000024-0.000073 g-eq per 1 g polymer, molecular weight is within 1,000,000-3,000,000. Method of producing said salt comprises adding to aqueous solution of polyacrylic acid an aqueous solution of lithium acetate, stirring reaction mixture for 10 minutes, adding aqueous solution of copper acetate, mixing for 100 minutes, drying obtained solution in a vacuum at temperature not higher than 50 °C. Partial double lithium-copper salt of polyacrylic acid is suggested as a haemostatic agent in form of aqueous solutions in ratio of 1-5 g of basic substance at 100 ml of water (1-5 %) and said agent has high antimicrobial and antifungal action during external application.

EFFECT: can be used in medical practice as a haemostatic agent of local action, for example when performing surgical operations, for stopping bleeding in traumatic wounds, in dentistry, simultaneously having bactericidal, fungicidal, bacteriostatic and fungistatic action.

3 cl, 5 ex, 1 tbl

Description

The invention relates to the creation of new chemical compounds that can be used in medical practice as topical hemostatic agents, for example, during surgical operations, to stop bleeding in traumatic wounds, in dentistry, etc.

On the basis of polyacrylic acid (PAA), which has some hemostatic effect due to the formation of interpolymer complexes (clots) with blood proteins, effective hemostatic agents, such as feracryl (SU 698622), argacryl (RU 2220982) and ciacryl (RU 2314815), are created. incomplete metal salts of iron, silver, zinc, respectively, as well as hemostatic agents based on incomplete salts of lithium (RU 2424813), sodium (RU 2428990), potassium (RU 2426546), rubidium (RU 2424814), cesium (RU 2428989) and double incomplete salt of lithium and zinc - lycicryl (RU 2424813). When using some metal salts of PAA as a hemostatic agent, an antiseptic effect and the regeneration of damaged tissues are also observed, for example, argacryl has a very strong antiseptic effect, Feracryl and Ciacryl, as well as Lycril, have slightly weaker effects. The creation of hemostatic antiseptics is very important, since very often bleeding is complicated by infectious infections, and the therapeutic effect of antibiotics, due to the resistance of pathogenic microflora to them, is very weak.

Therefore, the technical task of this invention is the development of new hemostatic agents based on PAA with significant antimicrobial activity.

The solution to this problem is achieved as follows: As a drug with these properties, a new incomplete double lithium-copper salt of polyacrylic acid (lycupril) is proposed, corresponding to the general formula: (-CH ₂ CHCOOH-) _n (-CH ₂ CHCOOLi-) _m (- CH ₂ CHCOOCu _0.5 ) _k , where n = 318871-956613; m = 96136-288408, k = 24573-73718, the content of Li and Cu is 4.43-4.46 and 3.93-4.10 0.00034-0.00101 g-at and 0.00029-0.00093 g-at, respectively, and the content of carboxyl groups is 0 , 000024-0.000073 g-equiv per 1 g of the polymer, the molecular weight is in the range from 1000000-3000000.

An incomplete double lithium-copper salt of polyacrylic acid (lycupril) is obtained by the interaction of aqueous PAA solutions with calculated amounts of lithium acetate first, and then after 10 minutes of copper acetate at room temperature. The reaction mixture is subjected to vacuum drying at a temperature not exceeding 50 ° C. The finished lycupril is a translucent polymer plate of blue color.

The hemostatic ability of lycupril was tested according to the standard method for stopping bleeding in Chinchilla rabbits using the example of an artificial liver wound with uniform application of an aqueous solution of lycupril at concentrations of 1, 2 and 5%.

The antimicrobial activity of lycupril was studied for both gram-positive and gram-negative bacteria, as well as its effect on fungal infection:

1. It has been experimentally established that lycupril in the form of a 1-5% aqueous solution exhibits a high hemostatic effect that is superior to that of Feracryl, Argacryl and Cyacryl,

2. Lycupril has bactericidal, fungicidal, bacteriostatic, fungistatic actions.

The following examples illustrate the invention.

Example 1. Synthesis of an incomplete double lithium-copper salt of polyacrylic acid (PAA)

10.0 g PAA with a molecular weight of 1,000,000-3,000,000 is dissolved with stirring in 250 ml of distilled water. An aqueous solution of 5.18 g of lithium acetate in 10 ml of water (which theoretically corresponds to the content of 5% lithium in the target product) is added to the solution with stirring at room temperature. The reaction mixture is stirred for 10 minutes, then an aqueous solution of 1.68 g of copper acetate in 100 ml of water (which theoretically corresponds to the content of 4% copper in the target product) is added to it, stirred for another 100 minutes. The resulting solution was dried in vacuum at a temperature not exceeding 50 ° C. The yield of solid substance in the form of blue plates of 15 g is practically quantitative.

Example 2. The effect of incomplete double lithium-copper salt of polyacrylic acid (PAA) on stopping bleeding

The influence of the hemostatic effect of the incomplete double lithium-copper salt of polyacrylic acid (PAA) was studied under laboratory conditions on chinchilla rabbits weighing 3-4 kg under thiopental anesthesia (6-8 ml of a 1.0% solution intravenously, and then up to 6- 10 ml of a 1.5% solution intraperitoneally). Laparotomy was performed using a longitudinal section along the white line of the abdomen. Intestine was taken out into the wound, limiting it with napkins moistened with warm saline and the front surface of the liver. Using a special limiting device, a superficial razor wound of the liver was applied with a sharp razor of about 1.5 cm ² and a depth of about 0.3 cm. Parenchymal bleeding was stopped by uniformly applying a 1% aqueous solution of an incomplete double lithium-copper salt of polyacrylic acid (PAA) in a dose respectively 6.7 mg / cm ² .

As a control, we stopped the bleeding by tightly pressing a tampon from a gauze napkin to the wound surface. The stop of bleeding occurred in the control for 264 ± 15 seconds. When applying an incomplete double lithium-copper salt of polyacrylic acid (PAA) at a dose of 6.7 mg / cm ² in 155 ± 32 seconds.

Example 3. The effect of incomplete double lithium-copper salt of polyacrylic acid (PAA) in stopping bleeding

The influence of the hemostatic effect of the incomplete double lithium-copper salt of polyacrylic acid (PAA) was studied under laboratory conditions on chinchilla rabbits weighing 3-4 kg under thiopental anesthesia (6-8 ml of a 1.0% solution intravenously, and then up to 6- 10 ml of a 1.5% solution intraperitoneally). Laparotomy was performed using a longitudinal section along the white line of the abdomen. Intestine was taken out into the wound, limiting it with napkins moistened with warm saline and the front surface of the liver. Using a special limiting device, a superficial razor of the liver was applied with a sharp razor about 1.5 cm ² and a depth of about 0.3 cm. Parenchymal bleeding was stopped by uniformly applying a 2% aqueous solution of an incomplete double lithium-copper salt of polyacrylic acid (PAA) in a dose respectively 13.4 mg / cm ² .

As a control, we stopped the bleeding by tightly pressing a tampon from a gauze napkin to the wound surface. The stop of bleeding occurred in the control for 264 ± 15 seconds. When applying an incomplete double lithium-copper salt of polyacrylic acid (PAA) at a dose of 13.4 mg / cm ² - in 134 ± 19 seconds.

Example 4. The effect of incomplete double lithium-copper salt of polyacrylic acid (PAA) in stopping bleeding

The influence of the hemostatic effect of the incomplete double lithium-copper salt of polyacrylic acid (PAA) was studied under laboratory conditions on chinchilla rabbits weighing 3-4 kg under thiopental anesthesia (6-8 ml of a 1.0% solution intravenously, and then up to 6- 10 ml of a 1.5% solution intraperitoneally). Laparotomy was performed using a longitudinal section along the white line of the abdomen. Intestine was withdrawn into the wound, limiting it with napkins moistened with warm saline; and the front surface of the liver. Using a special limiting device, a superficial razor of the liver was applied with a sharp razor about 1.5 cm ² and a depth of about 0.3 cm. Parenchymal bleeding was stopped by uniformly applying a 5% aqueous solution of an incomplete double lithium-copper salt of polyacrylic acid (PAA) in a dose respectively 33.5 mg / cm ² .

As a control, we stopped the bleeding by tightly pressing a tampon from a gauze napkin to the wound surface. The stop of bleeding occurred in the control for 264 ± 15 seconds. When applying an incomplete double lithium-copper salt of polyacrylic acid (PAA) at a dose of 33.5 mg / cm ² - for 146 ± 13 seconds.

Example 5. The effect of incomplete double lithium-copper salt of polyacrylic acid (PAA) on gram-positive, gram-negative and fungal infections.

Reference cultures of microorganisms recommended by GFII for determining the antimicrobial effect of the preparations, as well as freshly isolated clinical test strains of gram-positive and gram-negative microorganisms, were used as test strains.

The verification of bacteriostatic and biocidal action was carried out in accordance with the requirements of the "Guidelines for the experimental (preclinical) study of drugs" [1] and the State Pharmacopoeia [2]. For the preparation of microbial suspensions (suspensions), 20 hour agar cultures of test strains of microorganisms were used. Cultures were grown on weakly alkaline agar. The suspensions were densitometrically (Densimat Bio Merieux densitometer) adjusted to a density of 0.5 according to McFarland standard (containing approximately 1.5 × 10 ⁸ CFU / ml). A series of successive dilutions of 1:10 to a concentration of 1.5 × 10 ⁶ CFU / ml was prepared from the obtained microbial suspension. The specific antimicrobial activity of polymetalloacrylates was studied by the standard method of serial dilutions in liquid nutrient media (MPB, candida - Saburo broth). Before use, the preparations were dissolved in sterile distilled water based on the initial working concentration of 5%, 2%, 1% and intermediate dilutions of 1:10 were prepared. Various dilutions (quantities) of the studied drugs were introduced in an amount of 1 ml in test tubes with the appropriate nutrient medium in two replicates. Microbial loads were added in a volume of 0.1 ml to all tubes, including control. Bacteria were incubated in the thermostat for 24 hours, yeast for 48 hours. Test tubes containing no test samples served as control. Analysis was carried out first visually; determining the presence or absence of growth in a medium containing various concentrations of the test compound. The last test tube of a series with growth retardation (transparent broth) corresponds to the minimum inhibitory (bacteriostatic) concentration of the drug in relation to this strain. After an optimal incubation period for each microbial species, inoculations from all transparent tubes were seeded on solid nutrient media (Mueller-Hinton agar, Saburo) to determine cell viability. The lowest concentration of the substance in the test tube was noted, sowing from which did not give growth. This concentration was taken as minimal bactericidal. The results are presented in Table 1. The antimicrobial activity of the drug lycupril. Note: (-) - growth of test strains of microorganisms was not detected, (+) - growth of test strains of microorganisms was detected, (+ -) - weak growth.

In 100% of cases, a bactericidal action (MBC) has an incomplete double lithium-copper salt of polyacrylic acid (PAA) - 5% solution and its dilution 1:10 (0.5%), 2% and 1%; fungicidal - 5% and 2% solutions, bacteriostatic (MIC) in 100% of cases - 5% solution and its dilution 1:10 (0.5%), 2% and 1% solutions, fungistatic action - 5%, 2% solutions .

Information sources

1. Guidance on the experimental (preclinical) study of new pharmacological substances / Under the general editorship of R.U. Khabrieva. - 2nd ed., Revised. and add. - M.: 2005.

2. The State Pharmacopoeia. - XII ed. Part 1. - M .: Scientific center for the examination of medical devices, 2007. - 697 p.

Claims (3)

1. Incomplete double lithium-copper salt of polyacrylic acid, characterized in that it corresponds to the formula: (-CH ₂ CHCOOH-) _n (-CH ₂ CHCOOLi-) _m (-CH ₂ CHCOOCu _0.5 ) _k , where n = 318871-956613; m = 96136-288408, k = 24573-73718, the content of Li and Cu is 4.43-4.46 and 3.93-4.10 0.00034-0.00101 g-at and 0.00029-0.00093 g-at, respectively, and the content of carboxyl groups is 0 , 000024-0.000073 g-equiv per 1 g of the polymer, the molecular weight is in the range from 1000000-3000000. 2. A method of producing a compound according to claim 1, characterized in that an aqueous solution of lithium acetate is added to an aqueous solution of polyacrylic acid at room temperature, the reaction mixture is stirred for 10 minutes, then an aqueous solution of copper acetate is added, stirred for 100 minutes, the resulting solution is dried in vacuum at a temperature not exceeding 50 ° C. 3. A tool with an effective hemostatic effect and at the same time high bactericidal; fungicidal, bacteriostatic and fungistatic actions for external use, characterized in that it contains the compound according to claim 1 and is made in the form of solutions in water at a ratio of 1-5 g of the basic substance per 100 ml of water (1-5%)