WO2012154078A1

WO2012154078A1 - Antimicrobially active pharmaceutical composition for parenteral use, and method for producing said composition

Info

Publication number: WO2012154078A1
Application number: PCT/RU2011/000781
Authority: WO
Inventors: Виктор Львович ЛИМОНОВ; Константин Валентинович ГАЙДУЛЬ; Александр Валерьевич ДУШКИН
Original assignee: Limonov Viktor Lvovich
Priority date: 2011-05-11
Filing date: 2011-10-05
Publication date: 2012-11-15
Also published as: EA021847B1; EA201100736A1

Abstract

The invention relates to pharmacology, medicine and to the pharmaceutical industry, in particular to a method for producing original composite antimicrobial preparations for parenteral use, said preparations having increased therapeutic effectiveness in the treatment of severe forms of infectious inflammatory diseases. The proposed pharmaceutical composition comprises an antibiotic - glycopeptide and highly dispersed nano-structured silicon dioxide as the active substance. The claimed method for the production of the pharmaceutical composition consists in mixing the antibiotic - glycopeptide with the highly dispersed nano-structured silicon dioxide, characterized in that the mixture of the above-mentioned substances in ratios of 10:1 to 70:1 by weight, respectively, is subjected to mechanical processing by means of impact-abrading actions to increase the finely dispersed fraction weight portion. The mixture produced is used for preparing injection solutions.

Description

PHARMACEUTICAL COMPOSITION OF ANTIMICROBIAL ACTION FOR Parenteral use,

METHOD FOR ITS PRODUCTION

The invention relates to antimicrobial pharmaceutical preparations and technologies for their preparation and can be used in medicine for the treatment of infectious and inflammatory diseases, as well as in the pharmaceutical industry for the production of medicines.

Currently, to ensure the successful treatment of many infectious and inflammatory diseases (endocarditis, sepsis, infections of the bones and joints, infections of the lower respiratory tract, infections of the skin and soft tissues, etc.) caused by gram-positive microorganisms - Staphylococcus aureus and Staphylococcus epidermidis (including methicillin -resistant strains), Streptococcus viridians, Streptococcus pyogenes and Streptococcus pneumoniae (including penicillin-resistant strains), Enterococcus faecalis and others, in international medicine antibiotics-glycopeptides that have international non-pathogens are widely used ntovannye here - vancomycin, teicoplanin and others.

[12].

However, it should be noted that in some cases when the infectious process is caused by microorganisms resistant to antibiotic-glycopeptides to different degrees, monotherapy with these drugs is often not effective, which forces specialists to replace them with other drugs (usually expensive) or use them in high doses in combination with antibiotics from other groups, thereby significantly increasing the risk of unwanted side effects [3, 4, 5]. In this regard, the development of new approaches to significantly increase the antimicrobial activity and clinical efficacy of glycopeptide antibiotics is an urgent task of modern experimental pharmacology and practical medicine.

In recent years, it was found that the use of a variety of nanoparticles as carriers for the delivery of various antibiotics directly into the microorganisms and into the cells of the immune system (macrophages), in order to increase the concentration of these drugs in the area of infectious inflammation, and, accordingly, enhance their antimicrobial properties, and also for stimulation the antibacterial activity of macrophages and their additional recruitment into infected tissues is a promising direction in the development of original technologies and methods of antibiotic therapy [6, 7, 8, 9, 10, 1 1, 12, 13].

The essence of the invention lies in the fact that in order to enhance the therapeutic efficacy of antibiotic glycopeptides, it is proposed to use S1O2 (silicon dioxide) nanoparticles, which, differing in pharmacologically beneficial properties of biocompatibility, biodistribution, biodegradation and low toxicity (regardless of the severity of structure porosity), are capable of parenteral administration serve as a carrier of antibiotics for intracellular delivery to macrophages, which are concentrated in the foci of deerias observed in the lungs, liver, kidneys, spleen, lymph nodes, heart, skin, bladder and other mammalian organs (i.e., significantly increase the concentration of antibiotics in infected areas), as well as significantly increase the antimicrobial activity of these cells of the immune system (in in particular, by stimulating the production of nitric oxide), thereby significantly enhancing the therapeutic effect of antimicrobials in the treatment of infectious and inflammatory diseases [14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24].

The invention solves the problem of creating a pharmaceutical composition of antimicrobial action for injection based on the use of antibiotic glycopeptides and nanoparticles of silicon dioxide, which has increased therapeutic efficacy (compared to conventional glycopeptides, which are considered as a prototype in this invention) in the treatment of infectious and inflammatory diseases caused by gram-positive microorganisms.

The problem is solved in that a pharmaceutical composition of antimicrobial action for injection is proposed that contains an antibiotic glycopeptide and highly dispersed nanostructured silicon dioxide in a weight ratio of (10-70): 1.

The problem is also solved by the fact that the proposed method of obtaining a pharmaceutical composition of antimicrobial and actions for parenteral use, comprising mixing an antibiotic-glycopeptide with other components, according to which the antibiotic-glycopeptide in the form of lyophilized the powder is mixed with highly dispersed nanostructured silicon dioxide powder in a weight ratio of (10-70): 1, and the resulting mixture is subjected to mechanical treatment by impact-abrasion.

The therapeutic efficacy of the proposed pharmaceutical composition is improved if the resulting mixture is machined by impact-abrasion such that the proportion of highly dispersed nanostructured silicon dioxide having a size of -25 μm is at least 35%.

For the preparation of the pharmaceutical composition, antibiotic glycopeptides — vancomycin and teicoplanin — were used, provided by the Russian pharmaceutical manufacturer LLC ABOLmed. The highly dispersed nanostructured silicon dioxide (hereinafter: BHS1O2) was AEROSIL 200 (INN - colloidal silicon dioxide) used in pharmacy, manufactured by Evonik Degussa Corporation) (Germany), consisting of round-shaped silicon dioxide nanoparticles (average diameter 7 -40 nm), combined into irregularly shaped microparticles having sizes <100 μm.

The composition composition is based on the phenomenon of reversible sorption of antibiotic-glycopeptide molecules by nano- and micro-sized particles of BHS1O2, as well as a decrease in the size of BHSi02 microparticles upon mechanical activation of its mixtures with the antibiotic-glycopeptide substance by intense impact-abrasion mechanical effects.

The inventive method for producing the above pharmaceutical composition by mechanically activating a powdered mixture of antibiotic glycopeptide and BHS1O2 by intensive impact abrasion can increase the proportion of finely dispersed (.2 5 μm) particles of BHS1O2 on which antibiotic glycopeptide molecules are adsorbed and which are macrophaged [25].

For this, a mixture of the above substances in a weight ratio of antibiotic-glycopeptide: BHS1O2 equal to (10-70): 1 is subjected to mechanical activation by impact-abrasion until the weight fraction of the finely divided fraction is increased to no less than 35%.

Injected solutions for parenteral administration (by diluting it with any known by the method adopted for glycopeptide antibiotics), consisting of fine particles of BHSiCte with antibiotic glycopeptide molecules reversibly sorbed on their surface.

The introduction into the proposed composition of highly dispersed nanostructured silicon dioxide in the ratios antibiotic glycopeptide: BHS1O2 from 10: 1 to 70: 1 by weight, respectively, is determined by a combination of two factors: 1) with an increase in the content of BHS1O2 more than 10% by weight of the composition in laboratory animals when it is intravenous administration, intoxication phenomena are observed; 2) with a decrease in the content of BHS1O2 below 1.4% by weight of the composition, its therapeutic effectiveness (in particular, in the treatment of bacterial sepsis in mice) does not actually differ from the baseline effectiveness of the initial antibiotic glycopeptide.

To obtain the compositions, a mechanochemical approach was used, which consists in processing a mixture of solid components by intense mechanical stresses - pressure and shear deformations, which are realized mainly in various types of mills that carry out impact-abrasive effects on substances. A mixture of a solid substance of an antibiotic-glycopeptide and highly dispersed nanostructured silicon dioxide, taken predominantly in ratios from 10: 1 to 70: 1 by weight, is subjected to mechanical activation in ball mills. The method used to obtain mixtures allows one to significantly avoid the chemical decomposition of the antibiotic glycopeptide, to achieve complete homogeneity of the powder components in comparison with the preparation of mixtures by simple mixing of the components, or by evaporation of their solutions and, as a result, determines the high pharmacological activity of the pharmaceutical composition.

As a quantitative criterion for the minimum required dose of mechanical stress, it is convenient to use the method of granulometry of the suspension of the resulting composition. In this case, it is necessary that the mass fraction of particles with a size of not more than 5 μm is at least 35%. The mechanical processing of powdered mixtures is carried out in rotary, vibration or planetary mills. As grinding media can be used balls, rods, etc. Pharmacological tests of the obtained composition in laboratory animals (mice) showed that the claimed composition prepared by the claimed method has significantly increased therapeutic efficacy in the treatment of bacterial sepsis caused by Staphylococcus aureus, compared with the usual antibiotic glycopeptide.

Thus, the use of the claimed pharmaceutical composition and method for its preparation provides the following advantages:

1) a clinically significant increase in the effectiveness and quality of antimicrobial therapy of severe forms of infectious and inflammatory diseases, a decrease in mortality;

2) environmental safety, waste-free and low-cost pharmaceutical production technology. The invention is illustrated by the following examples.

Example 1. Obtaining a solid composition of the antibiotic-glycopeptide / BH8U ₂ . A mixture of antibiotic-glycopeptide and BHSi0 ₂ in weight ratios of 10: 1, 30: 1 and 50: 1 by weight is processed for 1, 2 and 4 hours in a rotary ball mill. The particle size distribution of aqueous suspensions of the obtained composition variants (Micro-Sizer 201 laser granulometer was used), as well as HPLC analysis of the content of antibiotics in them (in% of the initial substance) and the degree of antibiotic sorption (in%) by BHSi0 _2j particles are given in Table. one.

Table 1. The particle size distribution of aqueous suspensions, the degree of sorption and the content of antibiotics in various variants of the composition

Composition Composition Size and% Degree of sorption Antibiotic content of antibiotic particles of BHSi0 ₂ * particles of BHSi0 ₂ (%)

% <3% <5 (%)

μm μm

Original BHSi0 ₂ 0.4 6, 1 - -

Vancomycin: VN8Yu ₂ (10: 1), 19.8 36.7 41, 6 97 mechanical activation 1 hour

BaHK0MHUHH: BHSiO ₂ (30: 1), 21, 5 39.4 55.7 98 mechanical activation 2 hours

Vancomycin: BHSiO ₂ (50: l), 19, 1 38, 1 53.3 95 mechanical activation 4 hours Teicoplanin: BHSi0 ₂ (30: 1), 22.3 37.9 54.5 97 mechanical activation 2 hours

* - highly dispersed nanostructured silicon dioxide

As can be seen from the table. 1, the selected conditions for obtaining the proposed composition can be increased to the required value (at least 35% of the total weight) of the fraction of the finely dispersed fraction of BHSi0 ₂ (particle size -2 5 μm), while avoiding chemical decomposition of antibiotic glycopeptides.

Example 2. Determination of therapeutic efficacy of antibiotic glycopeptides and pharmaceutical compositions in the treatment of sepsis.

Vancomycin, teicoplanin and mechanically activated for 2 hours compositions consisting of a mixture of vancomycin or teicoplanin: BH8 ₂ in weight ratios of 30: 1 and 50: 1, respectively, were studied.

To determine the therapeutic efficacy of glycopeptide antibiotics and pharmaceutical compositions with BHSi0 _{2, we} used a modified experimental model of sepsis and a method of statistical processing of the obtained results (χ ² ) [26, 27, 28].

Microorganisms: Staphylococcus aureus (ATCC N ° 25923 F-49).

Animals: experiments were performed on hybrid male mice (CBA x C ₅₇ Black / ₆ ) CBFi in accordance with the "Rules for the Use of Experimental Animals" (Appendix to the order of the USSR Ministry of Health dated 12.08. 1977, Jfg 755).

Experimental sepsis model and treatment regimen

A suspension of the daily culture of Staphylococcus aureus at a dose of 10 ¹⁰ CFU / mouse was injected intravenously in mice in a volume of 0.5 ml. The control group of mice was injected phys. solution (0.9% sodium chloride solution) in a volume of 0.5 ml. A day after infection, mice were injected intravenously daily for 3 days with vancomycin and teicoplanin and three variants of the pharmaceutical composition (vancomycin or teicoplanin / VSHUg) at a dose of 50 mg / kg, diluted in 0.25 ml of phys. solution The control group of mice according to the same scheme was introduced physical. solution in a volume of 0.25 ml.

The effectiveness of antibiotic therapy was evaluated by the number of surviving mice on the 7th day after infection. The data obtained are presented in table. 2, reflect the results of four independent experiments performed for each study group.

Table 2.

Therapeutic efficacy of antimicrobial therapy of bacterial sepsis

* - in% and in absolute terms (number of survivors / number of infected animals).

** - in comparison with vancomycin.

*** - in comparison with teicoplanin.

As can be seen from the table. 2, the proposed pharmaceutical composition (antibiotic-glycopeptide / BHSiCh ₎ has significantly increased therapeutic efficacy (more than 1, 3 times), compared with conventional vancomycin and teicoplanin, in the treatment of sepsis in experimental animals caused by Staphylococcus aureus.

Thus, based on the data obtained, we can conclude that the proposed pharmaceutical composition of antimicrobial activity for parenteral use (antibiotic-glycopeptide / VN8YU ₂₎ has significantly enhanced therapeutic effect in the treatment of severe infectious vospalitelngh diseases, as compared with a conventional vancomycin or teicoplanin (prototypes of the invention). Literature

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Claims

Claim

1. A pharmaceutical composition of antimicrobial action for parenteral use, containing an antibiotic-glycopeptide as a therapeutic substance, characterized in that it is in the form of a powder for the preparation of injection solutions and contains highly dispersed nanostructured silicon dioxide in a weight ratio of antibiotic-glycopeptide: highly dispersed nanostructured silicon dioxide, equal to (10-70): 1.

2. A method of obtaining a pharmaceutical composition of antimicrobial action for parenteral use, comprising mixing an antibiotic glycopeptide with other components, characterized in that the antibiotic-glycopeptide in powder form is mixed with powdered highly dispersed nanostructured silicon dioxide in a weight ratio of antibiotic-glycopeptide: highly dispersed silica, highly dispersed nanostructured equal to (10-70): 1, and the resulting mixture is subjected to mechanical treatment by impact-abrasion.