EA029076B1 - Antimicrobial agent - Google Patents

Abstract

The invention is related to pharmacy, in particular, to medicinal products based on silver(I) complexes in the form of hydrogels that can be used for treatment of infected surface wounds and slightly exudating burns, bedsores, ulcers that do not heal for a long time, skin abrasions. The objective of the invention is provision of a novel, efficient, broad-spectrum antimicrobial agent for external application characterized by high stability during storage. The objective is attained by that an antimicrobial agent including a silver-containing substance (complex of silver(I) with 2-(4,6-di-tert-butyl-2,3-dihydroxiphenylsulphanyl)acetic acid), polyvinyl pyrrolidone, methyl cellulose and water for injections, additionally comprises 2-(4,6-di-tert-butyl-2,3-dihydroxiphenylsulphanyl)acetic acid and propylene glycol with the following proportion of components, wt.%: complex of silver(I) with 2-(4,6-di-tert-butyl-2,3-dihydroxiphenylsulphanyl)acetic acid: 0.25-1.00; 2-(4,6-di-tert-butyl-2,3-dihydroxiphenylsulphanyl)acetic acid: 0.03-0.10; propylene glycol: 5.0-20.0; polyvinyl pyrrolidone: 1.5-2.5; methyl cellulose: 3.0-4.0; water for injections: the balance.

Description

The invention relates to pharmacy, in particular to medicines based on silver complexes) in the form of hydrogels, which can be used to treat infected superficial wounds and burns with weak exudation, bedsores, nonhealing ulcers, abrasions. The objective of the invention is to obtain a new effective broad-spectrum antimicrobial agent for external use, characterized by high storage stability. The task is achieved by the fact that an antimicrobial agent comprising a silver-containing substance (complex of silver) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid, polyvinylpyrrolidone, methylcellulose and water for injection further contains 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid and propylene glycol in the following ratio of components, wt.%: Silver complex) with 2- (4,6di-tert-butyl-2,3 α-dihydroxyphenylsulfanyl) acetic acid: 0.25-1.00; 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid: 0.03-0.10; propylene glycol: 5.0-20.0; polyvinylpyrrolidone: 1.5-2.5; methylcellulose: 3.0-4.0; water for injection: the rest.

029076

The invention relates to pharmacy, in particular to medicines based on silver complexes (1) in the form of hydrogels, which can be used to treat infected superficial wounds and burns with weak exudation, bedsores, nonhealing ulcers, abrasions.

Silver compounds are known to exhibit antimicrobial activity [1]. Among the substances used in medicine, silver salts (silver nitrate), silver complex compounds (silver sulfadiazine, silver sulfathiazole) and colloidal silver (collargol, protargol) should be distinguished [2]. On their basis, creams (Argosulfan, Sulfargin, Silvaderm, Dermazin, Flamazin), solutions (Poviargol, Collargol, Protargol, Argovit, Argonika) and aerosols (Silvederm, etc.) [2, 3] are made. Currently, for the treatment of infected wounds, 1 -2% ointments containing a silver complex (1) with sulfonamides, sulfadiazine and sulfathiazole, are widely used. Upon contact with wound exudates, there is a slow dissociation of the silver complex (1) with the formation of silver ions and sulfanilamide. Of this class are active against Gram-positive, Gram-negative bacteria (including Rzeiboshopaz aegidtoza, EzsyepsYa dream Rgo1ei§ spp., 31aryu1osossi§ spp., K1e§1e11a spp.), As well as pathogenic and opportunistic fungi (including SapFba spr. and dermatophytes). The main side effects of these drugs are associated with the systemic effects of sulfonamides: impaired blood formation (agranulocytosis, aplastic and hemolytic anemia, thrombocytopenia, leukopenia), skin and allergic reactions, dyspeptic manifestations, hepatitis, hepatocellular necrosis, dysfunction of the central nervous system, toxic nephrosis, a respiratory tract.

The creation of a new silver-containing substance - a complex of silver (1) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid - made it possible to eliminate the above-listed side effects and expand the spectrum of the pharmacological activity of silver complexes [5, 6].

The closest to the claimed technical solution, selected as a prototype of the invention, is an ointment (hydrogel) containing silver complex (1) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid, polyvinylpyrrolidone and as an ointment base, cellulose ether selected from the group comprising methylcellulose, hydroxypropylmethylcellulose, and carboxymethylcellulose, as well as water for injection [7]. The ointment has a high antibacterial and antifungal activity (1.06-12.5 µg / ml in terms of the active ingredient). Hydrophilic ointment base - cellulose ether gel - provides a high level of release of silver-containing substance from the ointment.

However, a significant drawback of this tool, which largely limits the possibility of its large-scale production and use in medical practice, is the low stability of the silver-containing component in its composition. When storing the ointment, the active ingredient decomposes, leading to the formation of silver nanoparticles, which eventually aggregate and form coarse silver, which causes an uncontrolled change in the standardized quality indicators of the drug (decrease in pharmacological activity, appearance of phase heterogeneity and change in appearance).

The objective of the invention is to obtain a new effective broad-spectrum antimicrobial agent for external use, characterized by high storage stability.

The task is achieved by the fact that an antimicrobial agent comprising a silver-containing substance (silver complex (1) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid), poly-vinylpyrrolidone, methylcellulose and water for injection, additionally contains 2 (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid and propylene glycol in the following ratio of components, wt.%:

silver complex (1) with 2- (4,6-di-shch /? e / i-butyl-2,3- dihydroxyphenylsulfanyl) acetic acid 0.25-1.00 2- (4,6-di-7 yretya-butyl-2,3- dihydroxyphenylsulfanyl) acetic acid 0.03-0.10 propylene glycol 5.0-20.0 polyvinylpyrrolidone 1.5-2.5 methylcellulose 3.0-4.0 water for injections rest

The invention is illustrated FIG. 1 and 2, where in FIG. 1 shows the kinetic profile of the decomposition of AdL ₂ in various media (1 — DMSO; 2 — propylene glycol; 3 — propylene glycol with b addition), and FIG. 2 - the dependence of the model solution absorbance Ad ₂ in propylene glycol, after 60 min, by weight of the stabilizer L, added to the system (at the rate of 1.0 mg Ad _2).

The synthesis of the silver complex (1) (Ad ₂ ) is carried out according to the method described in [8]. Methylcellulose (MC) is used as an ointment base (TU2231-107-57684455-2003), which has a high biocompatibility and adsorbing capacity and is widely used in the production of gels. In order to stabilize the active silver-containing component of the ointment, 2- (4.6- 1 029076

di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid (E), the preparation of which is described in [9], and propylene glycol (PG) (99.9%, I8P / EP) is used as a solvent for ADB ₂ and B Polyvinylpyrrolidone (PVP) (molecular weight 25-350 kDa) is traditionally used to suppress aggregation of silver nanoparticles, which can be formed by applying an ointment on the wound surface.

Production of the proposed drug is carried out in aseptic conditions by performing the following operations (based on 100 g of ointment, table. 1):

1) swelling of MC in water: 3.0-4.0 g of MC pour 66.5 g of hot water for injection (80 ° C), cool to 20 ° C and mix until the polymer is completely dissolved;

2) PVP dissolution: 1.5-2.5 g of PVP are dissolved in 20.0 g of water for injection at a temperature of 20 ° С with stirring; the resulting mixture is combined with gel MC and evenly mixed;

3) dissolution of compounds Adk ₂ and B: 0.03-0.10 g of stabilizer B and 0.25-1.00 g of the complex AdB _{2 is} dissolved in 5.0-20.0 g of propylene glycol at 20 ° C in a dark place the location, if necessary using ultrasonic dispersion;

4) the introduction of the solution Adk ₂ and B into the ointment base: the solution obtained according to claim 3, with stirring, is gradually added to the mixture obtained according to claim 2;

5) packaging and storage: the ointment is packaged in dark plastic containers, hermetically sealed and stored in a refrigerator at a temperature of 4-6 ° C.

Table 1

The compositions of the proposed drug

The difference of the proposed drug from the previously known silver-containing ointments is illustrated by the examples below, which provide comparative data on the antimicrobial activity and stability of samples of ointments of different composition.

Example 1

The antimicrobial activity of ointment samples was determined by diffusion of the substance in a dense nutrient medium. Suspensions of daily agar cultures of test microbes in physiological solution were standardized to 10 ⁵ CFU / ml and seeded with a lawn on a dense nutrient medium; the wells in the agar were created with a sterile punch with a diameter of 5 mm and were filled with the sample (sterile tap water was added to the control wells). Crops were kept in a thermostat for 24 hours (for mushrooms — 48 hours) and after incubation the diameter (mm) of growth inhibition zones around the wells with samples was measured (no growth inhibition zone and the presence of growth inside the well were taken as 0.0 mm) [10] . The results of the study of the antimicrobial activity of the proposed drug and the prototype are presented in table. 2

table 2

Antimicrobial activity of the proposed drug and prototype

Sample Diameter of growth inhibition zones, mm E. soN ATSS 11229 5. aigeis ATSS 6538 R. Aegy & Posture ATSS 15442 S. a1Syap8 ATSS 10231 Azr. tag ATSS 16404 Prototype 22 23 nineteen 15 sixteen Composition 1 22 22 18 18 24 Composition 2 22 22 18 17 22 Composition 3 21 20 17 sixteen nineteen

* The error in measuring the diameter of the delay zones was 1 mm.

From tab. 2 it follows that the antibacterial activity of ointments of different composition with respect to gram-positive (й ос ос ос ос ов д), and (й й мотри) and gram-negative (с с Н Н Р Р бакт бакт сопостав) bacteria is comparable, but the proposed drug, more effective than the prototype, suppressed the growth of yeast (SapSaa aytotox), but the proposed remedy, more effective than the prototype, suppressed the growth of yeast (SapSaa aq). ).

Example 2

A weighed compound Adk ₂ weighing 1.0 mg was dissolved in 4.9 ml of DMSO or propylene glycol with the addition of 0.1 ml of water. The stabilizing effect of compound B was studied by dissolving its sample together with the complex in propylene glycol. After complete dissolution, the samples were placed in a temperature-controlled cuvette (37.0 ° C) and the change in the optical density of the solution with time was recorded at

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a wavelength of 500 nm (the device 8o1ag RV2201). In the process of decomposition of complex ₂ is formed Ad ligand colored oxidation product of L and silver nanoparticles, which eventually coagulates and causes a marked turbidity of the solution. As can be seen from FIG. 1, the decomposition of the LdB complex ₂ proceeds more slowly with the use of propylene glycol as a solvent (due to its lower solvating capacity compared to DMSO). Introduction to E connection system dramatically reduces the rate of decomposition of the complex Ld ₂ (Fig. 1, curve 4), the optimum stabilizer content is 0.1 mg per mg of the complex (Fig. 2).

To compare the stability of ointment samples of different composition, a stress test was performed [11], in which weighed 1 g of ointment weighing 1 g was placed in a thermostat (50 ° C). The decomposition of ointment samples was recorded by the appearance of pink coloration of the product of catechol ligand oxidation and black silver particles. A prototype sample was decomposed under the same conditions after 10 minutes. The tool of the claimed composition did not change its appearance for 120 minutes. Complete decomposition (sharp blackening) of the prototype sample and the claimed composition was observed after 25 and 240 minutes, respectively.

When stored in a dark place at a temperature of 2-8 ° C, the ointment remains stable during the observation period (more than 1 year).

Examples 3-7.

After 1, 3, 6, 9 and 12 months after the preparation of the ointments, their antimicrobial activity was controlled (inhibition of the growth of E.eH bacterium, Table 3), phase heterogeneity (average particle size, Table 4) and appearance (color of the ointment, table . five). Antimicrobial activity was established according to the procedure described in Example 1. The ointment color was determined with scattered light on a white background, and the particle size using a light microscope.

Table 3

The change in antimicrobial activity (diameter of growth inhibition E. Eon, mm) of the proposed drug and prototype over time

Sample Sample storage time, months one 3 6 9 12 Prototype 20 18 17 15 15 Composition 1 22 20 22 21 21 Composition 2 22 22 22 21 20 Composition 3 20 20 21 20 20

Table 4

The change in the average particle size (μm) of the proposed drug and prototype over time

Sample Sample storage time, months one 3 6 9 12 Prototype 20.5 40.2 49.1 52.0 50.0 Composition 1 1.8 2.0 1.8 2.4 2.2 Composition 2 1.5 1.6 1.6 1.8 1.9 Composition 3 1.2 1.5 1.4 1.6 1.8

Table 5

Change the appearance of the proposed drug and prototype over time

Sample Sample storage time, months one 3 6 9 12 Prototype light gray Gray dark grey the black the black Composition 1 white white white white white Composition 2 white white white white white Composition 3 white white white white white

As follows from the tables, over time, the prototype changes its color (Table 5) - the formation of coarse silver occurs, which entails an increase in the average particle size (Table 4) and a decrease in antimicrobial activity (Table 3). The inventive tool remains stable over the period of observation (12 months): there is no significant change in the antimicrobial effect, particle size and appearance.

Compound b is capable of stabilizing the LdB ₂ complex in an ointment due to the following factors: 1) its reducing properties [12], which prevent the ligand from oxidizing,

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part of the complex Ag ₂ , under the action of oxygen in the air during storage and inhibit the decomposition of the complex; 2) the creation, as a result of dissociation of b (pK _a = 4.1), of an acidic medium in which the oxidation of catechols slows down significantly [13]; 3) displacement of the equilibrium towards the starting substances (A§ ₂₎ ointment in the medium in accordance with the Le Chatelier principle, as the anion L ^- formed by the dissociation of the complex A§ _2.

The ointment is a homogeneous white elastic hydrogel, which is easily distributed and removed from the skin. The pH of the aqueous extract of the ointment is in the range of 5.3 to 5.5, which corresponds to the level of acidity of human skin in normal. The tool of the claimed composition meets the requirements for soft dosage forms for external use (stability, particle size, pH, microbiological purity) and can be produced in the pharmaceutical industry.

Thus, the stability of the active ingredient in the composition A§ ₂ ointments unlike the prototype provided by introducing into the preparation of 2- (4,6-di-tert-butyl-2,3-digidroksifenilsulfanil) acetic acid and propylene glycol at a certain component ratio ointment, while the proposed tool showed a high level of antimicrobial activity of a broad spectrum of activity.

Information sources.

1. Kharkevich D.A. Pharmacology / D.A. Kharkevich. - M .: GEOTAR - Media, 2008, p. 538-547.

2. Register of medicines of the Republic of Belarus [Electronic resource] / Ministry of Health of the Republic of Belarus. Center for Expertise and Testing in Health. - Minsk, 2016. Access mode: 1Shr: // \ y \ y \ y.gse11gu / geGyapk. - Access date: 10/20/2016.

3. State register of medicines [Electronic resource] / Ministry of Health of the Russian Federation. - Moscow, 2016. - Mode of access: 1Shr: //8g15.go5ti1/ggau.gi/SYAB8.a5r. \. Access date: 10/20/2016.

4. Reference Vidal "Drugs in Russia" [Electronic resource] / Vidal Rus. Moscow, 2016. - Access mode: LIR: //^№№.U1Ya1.gts. - Access date: 10/20/2016.

5. Patent 15000, IPC А61К 31/192, А61К 31/28, А61Р 31/04, А61Р 31/10, А61Р 31/22, publ. 02.28.11.

6. Patent No. 17021, IPC А61К 31/192, А61К 31/28, А61Р 31/10, publ. 12/30/12.

7. Patent No. 15481, IPC A61K 31/192, A61K 31/28, A61P 31/04, A61P 31/22, publ. 12/30/11.

8. Synthesis and antimicrobial activity of complexes of silver (1) and copper (11) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid / A.A. Chernyavskaya [et al.] // Chem. - farm. journals - 2006. V. 40, No. 8, p. 7-9.

9. Synthesis and antioxidant properties of some derivatives of alkylated pyrocatechol / Maslovskaya L.А. [et al.] // Journal of General Chemistry. - 1996. - № 11, p. 1893-1898.

10. Determination of the sensitivity of microorganisms to antibacterial drugs // Methodological guidelines MUK 4.2.1890 - 04. - M. Federal Center for Sanitary and Epidemiological Surveillance of the Ministry of Health of Russia. - 2004, p. 314-315.

11. Production of medicines. Tests of stability = Tolerance of the hellish crocodiles. Stabilized on us: TCP 431 - 2012 (02041). - Enter 03/01/12. - Minsk: Belarusian. state Institute of Standardization and Certification, 2012. - P. 5-15.

12. δίΙνοΓ (I) and Negasiop apot sotr1s \ aIop \\ ϊ11ι 51epsa11u Pibegeb 8i1Gig - soyasht Frépo1 böpuΐίνθδ / Ν.ν. Bodshow | e1 a1.] // Royuebgop. - 2005. - Wo1. 24. - R. 611-618.

13. Cherniavskaya, A. A. Complexation and redox - the interaction of Ad (1) with derivatives of spatially shielded diphenols and aminophenols / A.A. Chernyavskaya, V.N. Povalishev // Weight. Nat Acad. Navar Belara Ser. x1m navuk. - 2005. - № 5, pp. 132-134.

Claims (1)

CLAIM Antimicrobial agent for external use, including a silver-containing substance (silver complex (1) with 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid), polyvinylpyrrolidone, methylcellulose and water for injection, characterized by which additionally contains 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid and propylene glycol in the following ratio of components, wt.%: silver complex (1) with 2- (4,6-di- tert-butyl-2,3-dihydroxyphenylsulfanyl) -acetic acid: 0.25-1.00; 2- (4,6-di-tert-butyl-2,3-dihydroxyphenylsulfanyl) acetic acid: 0.03-0.10; propylene glycol: 5.0-20.0; polyvinylpyrrolidone: 1.5-2.5; methylcellulose: 3.0-4.0; water for injection: the rest. - 4 029076