RU2751163C9 - Macozinone-based pharmaceutical composition for treatment of tuberculosis including multi- and extensively drug-resistant forms thereof - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to the field of medicine and pharmaceuticals. The first object constitutes a gastroretentive swellable mucoadhesive tablet for controlled release of macozinone in the stomach and upper small intestine, constituting a combined system including a complex of macozinone or a pharmaceutically acceptable salt thereof in a therapeutically effective amount and cyclodextrin in form of granules coated with a layer of mucoadhesive polymer, wherein said layer includes a solubiliser, and pressed into a polymer matrix based on two hydrophilic soluble or one hydrophilic soluble and one insoluble swellable and mucoadhesive polymers, and an additional pharmaceutically acceptable excipient. The second object constitutes an application of the tablet for prevention and/or treatment of a disease caused by mycobacteria.

EFFECT: technical result of the invention is production of a gastroretentive tablet containing macozinone or a pharmaceutically acceptable salt thereof with modified release of an active substance with an increased relative bioavailability, providing long-term maintenance of a therapeutically significant concentration of an active substance in the body under conditions of a single oral administration, exhibiting high compliance and suitability for use in regimens of complex therapy of tuberculosis infection.

26 cl, 4 dwg, 11 tbl, 14 ex

Description

Technology area

The present invention relates to pharmaceutical compositions for the treatment of mycobacteriosis based on 2- [4- (cyclohexylmethyl) piperazin-1-yl] -8-nitro-6- (trifluoromethyl) -4H-1,3-benzothiazin-4-one (macosinone) more specifically, the present invention relates to a solid oral gastro-retentive dosage form of macosinone and its use for the treatment of mycobacteriosis.

State of the art

In recent decades, the problem of increasing the effectiveness of treatment of tuberculosis and other mycobacteriosis has acquired particular relevance in connection with an increase in the incidence rate both in Russia and throughout the world.

In terms of the burden of disease and mortality, tuberculosis (hereinafter TB) is undeniably the most important and complex threat to human health. According to WHO, in 2018, about 10 million cases of tuberculosis infection were registered in the world (Global tuberculosis report 2019, World Health Organization), which is one of the leading causes of death of infectious patients. In 2018, about 1.5 million deaths from tuberculosis infection were registered in the world, tuberculosis remains one of the 10 leading causes of death worldwide, while mortality rates are significantly higher (approximately 2 times) than that of widespread HIV / AIDS. ... Despite the gradual decrease in the total number of patients in the world, the social significance and danger of this disease remains very high, especially due to the increase in the prevalence of primary resistance of the pathogen strains to known drugs.

Over the past twenty years, drug-resistant tuberculosis has reached alarming levels. The situation is aggravated by the fact that the emergence and spread of new drugs after a while is regularly accompanied by the successive emergence of forms of mycobacteria resistant to them; the new stability is often "layered" on the previous one, as a result acquiring a multiple character. De novo infection with strains that have already acquired multidrug resistance is quite common. In the 1990s, there was a significant increase in the number of reported cases of tuberculosis caused by multidrug-resistant strains (MDR-forms) that have acquired resistance to the main first-line drugs isoniazid and rifampicin. According to WHO, in 2018, about half a million new cases with rifampicin-resistant tuberculosis were registered, 78% of which were multidrug-resistant). In the last decade, many MDR - M. tuberculosis strains began to acquire additional resistance mutations - already to second-line drugs, which led to the emergence and spread of a new type (class) of pathogen mutants, designated as extensively drug-resistant strains (XDR strains). In addition to isoniazid and rifampicin, M. tuberculosis XDR strains are also resistant to fluoroquinolones and to injectable aminoglycosides (Jassal M., Bishai WR. Extensively drug-resistant tuberculosis. -Lancet Infect Dis. 2009 Jan; 9 (1): 19- 30. doi: 10.1016 / S1473-3099 (08) 70260-3. Epub 2008 Nov 5). To date, more than 50 countries have reported the isolation of XDR-tuberculosis strains in the clinic, which makes it necessary and important to find new drugs for the treatment of diseases caused by both "traditional" drug-sensitive and resistant strains.

Due to the peculiarities of the pathogenesis of tuberculosis, the presence in the body of both actively dividing and dormant forms of mycobacteria, as well as taking into account the increase in the number of strains and variants of the pathogen resistant to certain drugs, currently in world practice, the main method of treating tuberculosis is the complex use several drugs, which increases the likelihood of achieving the appropriate therapeutic effect in the event that the pathogen strain acquires resistance to one of the drugs. The need for the simultaneous use of several drugs imposes restrictions on the dosage forms of such drugs that are acceptable for clinical use. Therefore, drugs of the first and second line are currently exclusively oral solid forms (tablets, capsules).

Due to the rapid spread in the world of strains of the causative agent of this disease, resistant not only to traditional drugs (isoniazid, rifampicin, salicylates and broad-spectrum antibiotics, including modern fluoroquinolones), but also to recently introduced into clinical practice drugs of the latest generations (delamanid, bedaquiline ), the public health need for new anti-tuberculosis compounds with a new mechanism of action is growing significantly (Balganesh, TS, PM Alzari, and ST Cole. Trends Pharmacol Sci, 2008. 29 (11): p.576-81.). Such agents may also find use in the treatment of other mycobacterial infections.

A promising way to increase the effectiveness of treatment of tuberculosis and other mycobacteriosis in recent years continues to be the search and creation of fundamentally new anti-tuberculosis drugs by screening new natural molecules or chemical modification of known structures. The world is actively searching for new active substances that suppress the development of pathogenic mycobacteria.

To date, scientific studies at the preclinical and clinical level have shown that a new class of chemical compounds, 2-amino-substituted-1,3-benzothiazin-4-ones, is very promising for the treatment of mycobacterial diseases in humans and mammals. Methods for the chemical synthesis of this new class of antimycobacterial compounds and their use for the treatment of mycobacteriosis are known from documents WO 2012066518 and WO 2016151011.

At the same time, the compound 2- [4- (cyclohexylmethyl) piperazin-1-yl] -8-nitro-6- (trifluoromethyl) -4H-1,3-benzothiazin-4-one (PBTZ169, MHN Makosinon) (Makarov V, Lechartier B, Zhang M, Neres J, van der Sar AM, Raadsen SA, et al. Towards a new combination therapy for tuberculosis with next generation benzothiazinones. EMBO Mol Med. 2014; 6 (3): 372 -83; Shi J, Lu J, Wen S, Zong Z, Huo F, Luo J, Liang Q, Li Y, Huang H, Pang Y. 2018. In vitro activity of PBTZ169 against multiple Mycobacterium species.Antimicrob Agents Chemother 62: e01314-18. https://doi.org/10.1128/AAC.01314-18.), which is currently one of the promising candidates for the development of a new anti-tuberculosis drug.

Compounds of this class have a unique mechanism of inhibition of mycobacteria - suppression of the activity of the enzyme decaprenyl-phosphoribose-2'-epimerase (DprE1), which is necessary for the synthesis of a component of the mycobacterial cell wall. This mechanism is highly specific for mycobacterial strains. Moreover, makosinone is active against all tested M. tuberculosis strains, including clinical isolates from patients with MDR and XDR forms of tuberculosis. The minimum inhibitory concentration in vitro for macosinone is in the range of 0.3-80 ng / ml for rapidly growing cultures, from 1 to 30 ng / ml for the entire M. tuberculosis complex, and at a concentration of 80-100 ng / ml, macosinone suppressed the development of all of tested XDR strains (In vitro activity of drug candidate PBTZ169, hydrochloride against extensively drug-resistant clinical strains of Mycobacterium tuberculosis.L.N. accelerated diagnosis and chemotherapy LN Chernousova et al. Bacteriology, 2017, volume 2, No. 1, pp. 25-34). In experiments on animals, a significant superiority of makosinone in therapeutic efficacy compared to isoniazid has also been proven, especially with prolonged (more than 2 months) courses of oral administration. Due to the unique specific mechanism of action against the bacterial pathogen, makosinon has a low level of toxicity for mammals, which is one of the conditions for newly synthesized promising drugs against mycobacteriosis, the treatment of which is carried out, as a rule, very long courses lasting several months.

However, macosinone, as a promising new pharmaceutical active ingredient, has some significant drawbacks that impede the development of an effective oral drug based on it.

Makosinon is practically insoluble in water, and at the same time is characterized by low lipophilicity (the substance is practically insoluble in most organic solvents, including hexane, heptane, chloroform). Macosinon is a weak base (pKa = 5.9) with a pronounced dependence of solubility on pH values: it has a very low solubility in water at pH 1-2 (not higher than 70-80 μg / ml) at higher pH values, it sharply decreases and at pH> 5, the substance loses its solubility and crystallizes, which prevents its transport through the intestinal cell membranes

As a result of the combination of these features of the physicochemical properties, makosinone has an extremely low oral bioavailability; in experiments on animals, the level of its bioavailability was recorded at a level of 1-2%.

Taking into account the listed features, makosinon should be classified as class IV pharmacologically active substances according to the generally accepted BCS classification, that is, substances with poor water solubility and low permeability through biological membranes.

But even within its class, macosinone has such essential features for pharmaceutical development as low lipophilicity, a narrow window of bioavailability in the gastrointestinal tract. In addition, makosinone has a relatively low (according to clinical studies, less than 60%) level of metabolic transformation in the body. Taking this into account, macosinone cannot be attributed to any of the 4 known classes of another international classification, widely used in pharmaceutical development, taking into account the distribution characteristics of the substance (BDDCS).

These features fundamentally affect the choice of general approaches to the design of an effective drug based on this promising and highly active compound.

It is well known from the technical field that substances of class IV according to the BCS classification always require the development of special techniques to ensure their effective use in the clinic, and it is almost impossible to predict in advance which of the many existing and known techniques to a competent and qualified specialist in this field will be the most promising. The end result of such a development is often a unique combination of an active substance and excipients, and the development of a dosage form of pharmacologically active substances of class IV according to the BCS classification is an extremely difficult and non-trivial task for a specialist (BCS class IV drugs: Highly notorious candidates for formulation development / Ghadi R , Dand N. // J. Control Release, 2017. - 28; 248: 71-95).

Thus, on the path of the introduction into practice and the fight against severe and deadly socially significant infectious diseases of the new promising anti-tuberculosis drug makosinon, which has a uniquely high bactericidal activity against MDR and XDR strains of M. tuberculosis, there are significant barriers due to a number of physical features. - the chemical properties of this substance, distinguishing it not only from other known anti-tuberculosis drugs, but also not described in such a combination for any other pharmacologically active molecules.

Therefore, there is a need to develop and create a new effective oral drug based on the compound makosinone with a higher bioavailability and corresponding to modern principles and schemes of complex therapy of tuberculosis infection.

Disclosure of invention

The objective of the present invention is the development and creation of a new and effective pharmaceutical oral composition (dosage form) of the active substance of macosinone (PBTZ169), characterized by a sufficient level of bioavailability and therapeutic efficacy with a predominantly single daily oral intake, which is promising for use in clinical practice for prevention and / or the treatment of infectious diseases in humans or animals, the causative agents of which are mycobacteria, in particular M. tuberculosis, and suitable for serial production on an industrial scale.

The technical result of this invention is the development and production of containing macosinone - 2- [4- (cyclohexylmethyl) piperazin-1-yl] -8-nitro-6- (trifluoromethyl) -4H-1,3-benzothiazin-4-one or its pharmaceutically an acceptable salt, in particular a hydrochloride salt, of a pharmaceutical composition in the form of a gastro-retentive tablet with a modified release of the active substance, with an increased relative bioavailability, providing, under single oral administration conditions, for a long time, up to 24 hours, maintaining a therapeutically significant concentration of the active substance in the body with high compliance and suitability for use in the schemes of complex therapy of tuberculosis infection. The technical result of the present invention is also the development of a technologically simple method for producing the specified dosage form (gastro-retentive tablet), suitable for scaling up to traditional pharmaceutical production equipment.

The specified technical result is achieved by the development and preparation of a gastro-retentive swellable mucoadhesive tablet for controlled release of macosinone in the stomach and upper part of the small intestine, which is a combined system comprising a complex of macosinone or a pharmaceutically acceptable salt thereof in a therapeutically effective amount and cyclodextrin in the form of granules, additionally coated with a layer mucoadhesive polymer and pressed into a polymer matrix based on at least two hydrophilic soluble and / or insoluble swellable and mucoadhesive polymers and at least one additional pharmaceutically acceptable excipient.

In particular embodiments, the macosinone or a pharmaceutically acceptable salt thereof is a fine crystalline powder with a particle size of 30 to 500 μm.

In particular embodiments, the pharmaceutically acceptable salt is a hydrochloride, hydrogen sulfate, ethanesulfonate, or methanesulfonate.

In particular embodiments, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin, 6-monodeoxy-6-monoamine-β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-γ-cyclodextrin and / or sulfobutyl-β-cyclodextrin. In more preferred embodiments, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.

In particular embodiments of the invention, the mucoadhesive polymer coating the granules is a soluble carboxymethyl cellulose with a degree of substitution of at least 0.7. In some embodiments, the mucoadhesive polymer layer further comprises a solubilizer. In other particular embodiments of the invention, the solubilizer is a medium molecular weight polyethylene glycol, a monoester of polyoxyethylated sorbitan and higher fatty acids. In particular embodiments, wherein the polyethylene glycol is PEG 6000 or PEG 8000. In some embodiments, the polyoxyethylated higher fatty acid monoester is Polysorbate 80.

In particular embodiments, the hydrophilic soluble mucoadhesive matrix polymer is a soluble cellulose derivative or high molecular weight polyethylene oxide with a molecular weight in the range of 1,000,000 to 4,000,000 daltons. In some embodiments, the cellulose derivative is hydroxyethyl cellulose or hydroxypropyl methylcellulose.

In particular embodiments of the invention, the insoluble mucoadhesive matrix polymer is a high polymer carbomer. In some embodiments, the carbomer is carbopol.

In particular embodiments of the invention, the additional pharmaceutically acceptable substance is a filler, binder, glidant, or lubricant.

In particular embodiments of the invention, the weight ratio of macosinone or a pharmaceutically acceptable salt thereof to cyclodextrin in the complex is from 0.5: 1 to 2: 1.

In particular embodiments of the invention, the content of the complex of macosinone or its pharmaceutically acceptable salt with cyclodextrin is 50-80% of the mass. based on the total weight of the tablet. In some particular embodiments, the amount of macosinone is 125-500 mg.

In particular embodiments of the invention, the tablet has an initial length of at least 20 mm and a width of at least 10 mm.

In particular embodiments of the invention, the tablet has an aqueous swelling index after 6 hours of 95-150%.

In particular embodiments of the invention, the tablet according to the invention has an in vitro adhesion force in 0.01 N hydrochloric acid solution, which ensures its retention on the mucosal surface for at least 5 hours.

In particular embodiments of the invention, the tablet has a crushing strength after tabletting of 190-210 N.

In particular embodiments of the invention, the tablet is characterized in that the release rate of macosinone in vitro in 0.01 N hydrochloric acid solution is 25-50 mg / h.

In particular embodiments of the invention, the tablet is characterized in that the release of the active substance in vitro in the dissolution medium of 0.01 N HCl after 6 hours is 30-50%.

The specified technical result is also achieved through the use of a gastro-retentive swelling mucoadhesive tablet according to the invention for the prevention and / or treatment of a disease caused by mycobacteria.

In some embodiments, the mycobacteria are M. tuberculosis.

In particular embodiments of the invention, the disease is tuberculosis. In some embodiments, the tuberculosis is in multiple and / or extensively drug resistant forms.

In particular embodiments of the invention, the tablet is administered orally to the subject once a day.

In particular embodiments of the invention, the subject is a human.

The invention also includes a method for preparing a gastro-retentive tablet according to the invention, comprising three steps:

a) obtaining granules of macosinone or its pharmaceutically acceptable salt in the form of a complex with cyclodextrin;

b) the inclusion of the granules obtained in stage a), in the composition of the secondary granules with their coating with a layer of a mixture of a bioadhesive agent and a solubilizer;

c) pressing dry secondary granules obtained in step b) into a tablet matrix based on one highly swellable and one strongly mucoadhesive polymer.

Definitions and terms

Various terms relating to the objects of the present invention are used above and also below in the description, in the claims and the abstract. Unless otherwise specified, all technical and scientific terms used in this application have the same meaning as understood by those skilled in the art. References to techniques used in describing the present invention refer to well known techniques, including modifying these techniques and replacing them with equivalent techniques known to those skilled in the art.

In the description of this invention, the terms "includes" and "including" are interpreted as meaning "includes, among other things". These terms are not intended to be construed as “consists only of”.

Macosinon. The chemical compound macosinone (or PBTZ169) is 2- [4- (cyclohexylmethyl) piperazin-1-yl] -8-nitro-6- (trifluoromethyl) -4H-1,3-benzothiazin-4-one, (CAS 1377239- 83-2) characterized by the following structural formula:

(gross formula - C ₂₀ H ₂₃ F ₃ N ₄ O ₃ S, molecular weight 456.5) as well as its pharmaceutically acceptable salts, hydrates, complexes, polymorphic variants. Macosinone (calculated as base) in the compositions according to the invention may be present in amounts from 125 to 1000 mg, preferably from 250 to 500 mg.

Macosinone hydrochloride (PBTZ169 HCl) (2- [4- (cyclohexylmethyl) piperazin-1-yl] -8-nitro-6- (trifluoromethyl) -4H-1,3-benzothiazin-4-one hydrochloride) has the structural formula:

gross formula - C ₂₀ H ₂₄ ClF ₃ N ₄ O ₃ S, molecular weight 492.5; the properties of the compound are shown in more detail in Table 1.

The term modified release tablet in this document describes a composition (dosage form) characterized, after ingestion, by a sustained and controlled release of the active substance in a given area of the gastrointestinal tract, mainly in the stomach and upper small intestine, as opposed to immediate release dosage forms, which after ingestion uncontrollably and immediately occurs directly in the stomach.

The term pharmaceutically acceptable means a material, a component that is biocompatible with the human body. As used herein, the term pharmaceutically acceptable salt refers to those salts that, within the scope of medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reaction, etc., and meet a reasonable balance of benefit and risk. ... Some examples of the salts according to the present invention, in particular, are hydrochloride, hydrogen sulfate, ethane sulfonate or methanesulfonate.

The term therapeutically effective amount means that amount of a compound that, when administered as mono- or combination therapy, is sufficient to prevent and / or treat infectious diseases in humans or animals caused by mycobacteria.

The term dosage in this document means the content of the active (active) substance in quantitative terms per unit volume or unit of mass in accordance with the dosage form, namely, a unit amount of the active ingredient in one unit of the dosage form, for example, in a tablet.

Pharmacokinetic curve - a curve reflecting the change in time of the concentration of an active substance or its metabolite in blood plasma or any organ or tissue in the body. The characteristics of the pharmacokinetic curve used in the present invention are indicators:

AUC is the area under the curve reflecting the change in time of the concentration of the active substance or its metabolite in the blood plasma or any organ or tissue in the body in a certain period of time after drug administration;

T _max is the moment recorded during the test from the injection of the drug to the maximum concentration in the blood plasma (biological body fluid);

T _1/2 - the moment of time from the beginning of the test, when the registered maximum concentration of the substance in the blood (biological body fluid) decreases by 2 times;

MRT is the duration of existence (retention) of a drug substance or metabolite in the body calculated from registered pharmacokinetic parameters;

F _rel is a parameter determined by comparing AUC indicators that describes the degree of bioavailability of a medicinal substance for the dosage form used in the study in comparison with another dosage form tested under the same conditions.

Cyclodextrins (CD) are a class of cyclic oligosaccharides obtained by enzymatic cleavage of starches and consisting of 6 (α-cyclodextrins, α-CD), 7 (β-cyclodextrins, β-CD) or 8 (γ-cyclodextrins, γ-CD) residues α-D (+) - glucopyranose (dextrose) linked into macrocycles by α-D-1,4-glycosidic bonds in such a way that the cyclodextrin molecule has the shape of a truncated cone with a hydrophobic inner cavity. When starch is treated with the microbial enzyme cyclodextring glucanotransferase, so-called large-ring cyclodextrins can also be obtained, containing nine, ten, eleven and more (up to 30-60) glucose residues in the cycle - the so-called δ-cyclodextrins, ε-cyclodextrins, ξ -cyclodextrins, etc.). Structural formulas of the three main types of cyclodextrins (α-CD, β-CD, γ-CD). The conical shape of the cyclodextrin molecule is stabilized by hydrogen bonds between OH groups, as well as by α-D-1,4-glycosidic bonds. All OH groups in cyclodextrins are located on the outer surface of the molecule. Therefore, the inner cavity of cyclodextrins is hydrophobic and is capable of forming inclusion complexes with various molecules of organic and inorganic nature in aqueous solutions. In inclusion complexes, the cyclodextrin ring is a "host" molecule, the incorporated substance is called a "guest".

Inclusion complexes in aqueous solutions dissociate into cyclodextrin and the starting substance, showing the basic properties of the latter. When heated above 50-60 ° C, the complexes usually disintegrate completely and usually restore their structure upon cooling.

The cyclodextrin suitable for the preparation of the pharmaceutical composition according to the invention can be, in particular, selected from 2-hydroxypropyl-p-cyclodextrin, 6-mono-deoxy-6-monoamine-β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-γ-cyclodextrin dextrin, sulfobutyl-β-cyclodextrin or mixtures thereof, while the quantitative ratio of macosinone: cyclodextrin in the claimed pharmaceutical composition can be from 0.5: 1 to 1: 5.

Mycobacteriosis is an infectious disease of humans or animals caused by pathogenic strains of the genus Mycobacterium (Mycobacterium). Within the framework of the present invention, it can be caused, in particular, by one or more pathogens selected from Mycobacterium tuberculosis (the causative agent of human and animal tuberculosis), Mycobacterium bow's, Mycobacterium kansasii, Mycobacterium scrofulaceum, Mycobacterium aviumintracelluluare complex, Mycobacterium lepraelosis or Mycobacterium lepraelosis.

MDR strains are multidrug-resistant strains of the causative agent of tuberculosis that have acquired resistance to first-line drugs in accordance with the WHO clinical guidelines.

XDR strains are extensively drug-resistant strains of the causative agent of tuberculosis that have acquired, in addition to the MDR characteristics, resistance to drugs of the second and subsequent lines of therapy according to the WHO clinical guidelines.

A pharmaceutical composition is a combination of an active active (medicinal) substance and pharmaceutically acceptable excipients that ensure the creation of a medicinal product.

Medicinal product is a pharmaceutical composition in a specific dosage form that provides the use of a medicinal substance to achieve the desired therapeutic effect.

Matrix - polymer base of a modified release tablet. The matrix can be based on hydrophilic (hydrophilic matrix) and / or hydrophobic polymers, which can be water-soluble or insoluble.

A gastro-retentive tablet is a dosage form in the form of a tablet, which, due to the design features, can linger in the stomach for a long time.

Monolithic delivery system (tablet) - in the framework of this invention, a monolithic (uniform) delivery system (tablet) is understood as a tablet obtained by simultaneous effective mixing of an active active substance (pharmaceutical substance) with the components that form the tablet matrix, followed by tabletting the resulting mixture to form systems with components (substances) evenly distributed in it.

The examples presented below relating to the preparation of a pharmaceutical composition for the treatment of mycobacterial infection, in particular due to Mycobacterium tuberculosis, in accordance with the present invention, as well as to the method for their preparation, do not limit the scope of the claims, but are given only to illustrate the present invention.

Brief description of the drawings.

Figure 1. Dependence of the solubility of the hydrochloride of makosinone HCI (μg / ml) on the pH values of the medium.

Dissolution media with pH 1.2, 4.5 and 6.8 - in accordance with the State Pharmacopoeia or the European Pharmacopoeia for testing the dissolution of dosage forms. The temperature of the solutions is 37.5 ° C. Tests in an apparatus of the "paddle mixer" type (apparatus No. 2), rotation speed - 150 rpm; the volume of the dissolution medium is 1000 ml.

Figure 2. Target cumulative release curve of macosinone from a modified release gastro-retentive tablet for a 500 mg macosinone tablet. The fill marks the permissible range of variation of the experimentally recorded indicators.

Figure 3. Time release of macosinone HCI from an immediate release dosage form - hard gelatin capsules, dosage 80 mg (reference drug according to example 9), in solutions with different pH values.

Dissolution media with different pH values - in accordance with the State Pharmacopoeia or the European Pharmacopoeia for testing the dissolution of dosage forms. The temperature of the solutions is 37.5 ° C. Tests in an apparatus of the "paddle mixer" type (apparatus No. 2) rotation speed - 150 rpm; the volume of the dissolution medium is 1000 ml.

Figure 4. Curves of changes in the content of makosinone in the blood plasma of animals when testing various oral dosage forms. Tests on dogs of the Beagle breed (5 dogs per group) under conditions of oral administration of drugs).

The ordinate is the concentration of makosinone in the blood plasma, ng / ml; the abscissa is the time after administration, hours.

A - Curves based on actual data. B - calculated curves of changes in the content of makosinone in the target organ (lungs), taking into account experimental data on tissue bioavailability (retention of the active substance in organs and tissues of animals)

The horizontal dashed line (7 and 8, respectively) marks the calculated minimum threshold for therapeutically effective concentration

Curve symbols:

1, 4 - immediate release preparation - hard gelatin capsules, dose 480 mg, calculated, 6 capsules with a dose of 80 mg, (reference drug, example 9);

2, 5 - gastro-retentive tablet of composition GR10, 500 mg (tablet according to the invention, example 6);

3, 6 - gastro-retentive tablet of composition GR20, 500 mg (reference preparation, example 11).

Detailed disclosure of the invention.

The present invention solves a complex problem, namely, the invention is directed to the development of an effective drug (pharmaceutical composition in a specific dosage form) based on macosinone hydrochloride for oral administration for the treatment of infectious diseases caused by mycobacteria, in particular M. tuberculosis (including resistant forms), intended for a single dose during the day, and such a composition should ensure the maintenance of a therapeutically effective concentration of the specified active substance in the body during the period between two successive doses of the drug.

The problem of developing an effective drug for the treatment of tuberculosis (including its resistant forms) is extremely urgent. Moreover, at present, tuberculosis therapy involves treatment with several drugs at the same time ("cocktail" of drugs), therefore, along with the effectiveness and bioavailability, the compliance of the drug (suitability and acceptability for patients and medical personnel) is also an important factor when used in a clinical hospital in combination with several drugs with different pharmacokinetic and pharmacodynamic properties.

From the prior art, the basic principle of treatment of forms of tuberculosis caused by MDR / XDR strains of pathogens is known, based on the simultaneous administration of several drugs with the exclusion of ineffective drugs and (or) drug combinations from the scheme. Due to the constant increase in the share of mycobacterial strains that have acquired resistance to traditional drugs in the world, in recent years, active development, testing and introduction into clinical practice of new active molecules that effectively suppress the growth and development of drug-resistant pathogenic bacteria have been carried out in many countries.

In recent years, two new active anti-tuberculosis compounds have been registered and used in a number of countries around the world, which, according to the International Recommendations for the Treatment of Drug-Resistant Tuberculosis Treatment of 2019 (WHO consolidated guidelines on drug-resistant tuberculosis treatment / 2019 / ISBN: ISBN 978-92-4 -155052-9) recommended as effective drugs for this purpose - delamanid and bedaquiline. However, by now, the effectiveness of the use of new drugs has begun to decline. In particular, to date, clinical and laboratory data are accumulating that the frequency of mutations in resistance to delamanid (which has been actively used in the treatment of tuberculosis since 2014) has already exceeded the frequency of mutations in resistance to rifampicin and has approximately reached the same level as mutations to isoniazid. As shown by special epidemiological studies, in populations of patients with tuberculosis who have not received the new anti-tuberculosis drug bedaquiline that has recently entered the markets of developed countries, up to 6-7% of patients already carry strains that are resistant not only to bedaquiline, but also cross-resistant to one standard drug from the combined treatment regimen - clofazimine.

Moreover, the peculiarities of the physicochemical properties and the related pharmacokinetics of bedaquiline (extremely long, for almost 5 months after the end of administration), a slow decrease in its concentration in tissues aggravates the problem of spontaneous selection in the body of variants of the pathogen resistant to this drug.

The disadvantages of bedaquiline can also include the presence of restrictions on the dosage of tablets and the need to simultaneously take 4 tablets (with a dosage of 100 mg), which impairs its compliance in complex therapy.

In addition to these problems with the increasing accumulation of resistant strains of mycobacteria in the population, the disadvantages of delaminade can also include the need to take the drug twice a day.

The recently synthesized new class of chemical compounds - benzothiazinone derivatives, new in terms of the mechanism of antimicrobial action on mycobacteria (suppression of cell wall synthesis) - is currently being considered as a more promising candidate for the role of a drug for inclusion in the treatment regimen of drug-resistant forms of tuberculosis.

In particular, makosinone (PBTZ169) is characterized by a significantly higher efficiency of suppression of all tested XDR variants of the pathogen (the minimum inhibitory concentration (MIC) of makosinone is 80-110 ng / ml, while for bedaquiline it is about 500-600 ng / ml).

However, the study of the physicochemical properties of this new class of chemical substances revealed the unique characteristics of these compounds, which significantly distinguish them from the overwhelming majority of known drugs and significantly complicate the development of dosage forms suitable for clinical use on their basis.

In particular, macosinone was found to be practically insoluble in water (Piton J, VocatA, Lupien A, Foo CS, Riabova O, Makarov V, Cole ST. 2018. Structure-based drug design and characterization of sulfonyl-piperazine benzothiazinone inhibitors of DprE1 from Mycobacterium tuberculosis Antimicrob Agents Chemother 62: e00681-18. https://doi.org/10.1128/AAC.00681-18). Even the base salts of these substances, such as makosinone hydrochloride, chosen as the most acceptable potential candidates, were also practically insoluble in most polar and non-polar solvents. At the same time, this compound has a relatively low lipophilicity, is characterized by a sharp decrease in the already extremely low solubility in water in the zone of neutral pH values, and, as a consequence, by low permeability through the membranes of intestinal epithelium cells, which does not depend on the presence of proteins or glycoproteins - carriers. By the combination of these properties, such compounds belong to the most problematic class IV for pharmaceutical development according to the international classification BCS (Biopharmaceutics Classification System (BCS) -based biowaiver applications: anti-tuberculosis medicines WHO Prequalification of Medicines Program Guidance Document February 2009)

And taking into account the sharp decrease in solubility characteristic of weak bases, which include makosinone, when going from acidic pH values to neutral ones (Fig. 1), it additionally belongs to a group of substances that are not numerous in this class with a very narrow window of bioavailability in the gastrointestinal tract. ...

The noted features of the physicochemical properties make this compound also unusual in terms of the combination of pharmacokinetic characteristics of ADME (absorption, distribution, excretion and metabolism). After being absorbed into the body after oral administration, makosinone undergoes metabolic transformation in the liver with the formation of a number of metabolites, the antimicrobial activity of which is significantly inferior to the activity of the parent molecule. Effectively binding to blood plasma proteins, makosinone, however, is rapidly eliminated from the body, mainly through the liver (with bile). After oral administration into the body of animals and humans, makosinon is characterized by very "fast" pharmacokinetics: the maximum content in the blood is reached on average 45-90 minutes after administration, and a decrease in concentration below therapeutically effective occurs within 6-8 hours. The known macosinone metabolites possessing "residual" bactericidal activity have similar pharmacokinetics.

Given the extremely low solubility and, at the same time, the relatively low intensity of metabolic transformation (less than 70% of the substance entered the body), makosinone actually cannot be attributed to any of the known classes of substances according to the international classification BDDCS (usually used in pharmaceutical development and taking into account the leading parameters of solubility and the degree of metabolic transformation) (The biopharmaceutics classification system (BCS) and the biopharmaceutics drug disposition classification system (BDDCS): Beyond guidelines / A. Charalabidis et al. - International Journal of Pharmaceutics, V. 566, 2019, P. 264-281 ).

The result of the combination of these unique properties of makosinone is its extremely low bioavailability when taken orally (no more than 2-4%).

The specified combination of the characteristics of the active substance makes the above-formulated problem of the present invention extremely difficult and non-trivial,

As a rule, class IV drugs with such low solubility and permeability indicators are used in the clinic in the form of dosage forms for parenteral administration (injection or infusion drugs), such as the antibiotics carbopenems used in the treatment of tuberculosis. However, taking into account the complex and complex regimen of tuberculosis therapy, especially its drug-resistant forms, the use of injectable drugs was recognized as low compliant and such drugs were classified as reserve drugs (used when 4-5-component therapy of resistant forms is ineffective).

The first-line drugs (ethambutol, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentin) in all modern regimens of tuberculosis therapy are oral solid dosage forms (tablets or capsules). Tablet dosage forms are more preferable (they make up 96.2% of respondents' preferences) due to the duration of taking this group of drugs (Global drug facility product catalog / Global Drug Facility. Stop TV Partnership. World Health Organization, Geneva 2016; Anti-tuberculosis dosage forms : assortment, main advantages, prospects for technological improvement / ME Kim et al // Pharmacy and Pharmacology No. 3, 2016. - pp. 38-55).

The process of developing an effective oral dosage form of makosinone, which is a consequence of its very low bioavailability and non-optimal pharmacokinetic profile, significantly complicates the need to use high dosages of the active substance in the final preparation. A therapeutically significant positive effect when tested in a clinic with the participation of tuberculosis patients was achieved only at a dosage level of the order of 640-1280 mg per patient. Considering that the maximum acceptable (taking into account the dissolution parameters set by the EP, USP, Russian GF XIV pharmacopoeias) dosage per unit of the developed solid dosage form of immediate release (capsules) was only 80 mg of the active ingredient, it becomes clear that the use of such dosage forms in real clinical practice becomes impossible due to their obvious non-compliance.

The present inventors have attempted to achieve a significant increase in the solubility of makosinone through such known conventional approaches as:

- the use of its various salts (of which macosinone hydrochloride had the best performance);

- additional crushing of crystals;

- addition to powders of known surfactants, such as sodium lauryl sulfate, tweens, Triton X-100.

However, all these techniques did not lead to any significant increase in the solubility of the substance.

In the same experiments, it was shown that the solubility of makosinone HCl does not change when the particle size in the substance powders changes in the range from 0.04 to 0.8 mm, obtained by sieve analysis. At the same time, fine-crystalline powders of the pharmaceutical substance of macosinone hydrochloride are distinguished by unsatisfactory technological characteristics, extremely low flowability and, at the same time, a high tendency of particles to aggregate and adhere to equipment surfaces. Of the surfactants tested, to an insignificant extent (no more than 10-15%), the dissolution rate of the substance of macosinone hydrochloride improved, mainly due to a decrease in the ability of crystals to aggregate, when small amounts (up to 0.005-0.01%) were introduced into solutions, tertiary ammonium detergents (for example, benzethonium chloride), but the use of such substances in pharmaceutical compositions is limited by the additional adverse side effects of such surfactants.

As noted above, the limiting factors that reduce the therapeutic efficacy of makosinone are not only its low solubility in water and low permeability through membranes, but also a narrow "bioavailability window" (a sharp decrease in solubility when the substance moves from the acidic environment of the stomach to the lower parts of the gastrointestinal path) and the associated insufficient duration of the absorption period. It should also be borne in mind that makosinon in the body quickly and effectively binds to blood plasma proteins and is unevenly distributed over organs and tissues, with a predominant delay in the lungs, spleen, and liver.

A negative point in this regard is the relatively fast metabolism found for makosinone already during the initial passage of the drug through the portal circle. At the same time, the detected and identified metabolites are significantly less active against the pathogen, which further reduces the therapeutic efficacy of the drug.

A consequence of the noted features of the process of bioabsorption and distribution is clearly suboptimal is the pharmacokinetic profile of the drug intended primarily for the treatment of chronic forms of mycobacterial infections, in which it is important to ensure a long period of maintaining an effective suppressing bacteria concentration in organs and tissues.

For clinical studies, the present inventors have developed an immediate release dosage form in the form of a hard gelatin capsule containing granular macosinone powder (example 9). However, in the course of the research, the low suitability of this form of the drug for clinical use was revealed. So, according to the results of clinical pharmacokinetic studies of this dosage form (capsules with a dosage of 80 mg) with the participation of volunteers, it was shown that the time to reach the maximum concentration in the blood plasma of this substance, taken once peros in dosages of 320, 640 g, and 1280 mg is 1 -2 hours, half-life is 10-12 hours. In the range of increasing dosages above 320 mg per day, the pharmacokinetics becomes non-linear and the use of dosages above 1280 mg becomes impractical, given the increasing risk of undesirable side effects. Preliminary estimates based on the results of a study of this oral dosage form on volunteers showed that the bioavailability of the active substance does not exceed 4-5% (calculated) even when dosages of 640 mg (8 capsules) or 1280 mg per person are reached.

In clinical studies of the 2nd phase with the participation of patients with uncomplicated tuberculosis, it was shown that positive therapeutic effects after a 2-week course of therapy were recorded (and not in all patients in the cohort) only at dosages of the drug not less than 640 mg.

The noted features of the pharmacokinetics of the active substance make it necessary to administer the drug with a frequency of at least two or even three times a day, which reduces its compliance in hospital therapy, increases the cost of course treatment, and, taking into account the need to use high single doses of the drug, also increases the risk of side effects. effects. A further increase in a single and course dose for immediate-release drugs with such a dosage becomes unacceptable due to obvious incompatibility, taking into account the mandatory complex nature of anti-tuberculosis therapy (mandatory simultaneous combination of 3 or 4 oral drugs of different classes).

The inventors of the present invention have surprisingly found that all of the above disadvantages and limitations are overcome by the development of a gastro-retentive swellable mucoadhesive tablet for the controlled release of macosinone in the stomach and upper small intestine, which is a combined system comprising a complex of a therapeutically effective amount of macosinone or a pharmaceutically acceptable salt thereof with cyclodextrin in the form of granules coated with a layer of mucoadhesive polymer and pressed into a polymer matrix based on at least two hydrophilic soluble and / or insoluble swelling and mucoadhesive polymers and at least one additional pharmaceutically acceptable excipient. As a result of the studies carried out, it was unexpectedly found that this dosage form of macosinone (gastro-retentive tablet) is not only characterized by increased bioavailability, but is also able to maintain a concentration effective against M. tuberculosis bacteria and an effective therapeutic effect of macosinone on cells of the causative agent of tuberculosis in the human body under conditions once a day of admission.

Examples of implementation of the invention

The examples given in the description do not exhaust the entire list of pharmaceutical compositions and methods for their preparation, which were made in the process of pharmaceutical development of a gastro-retentive tablet. In addition to the pharmaceutical compositions according to the described invention, examples are also given of the immediate release dosage form (capsule) used in experiments on the study of the pharmacokinetics, as well as several pharmaceutical compositions of gastro-retentive tablets, which differed from the tablets according to the invention according to the basic principles of design.

Example 1. The example describes the results of a study of the dependence of the solubility of macosinone hydrochloride on the pH values of the medium.

Figure 1 shows the dependence of the solubility of makosinone HCl (μg / ml) on the pH values of the medium. The dissolution medium is 0.01 N HCl. The volume of the dissolution medium is 1000 ml. The temperature of the solutions was 37.5 ° C. The tests were carried out in an apparatus of the "paddle stirrer" type (apparatus No. 2), rotation speed - 150 rpm.

From these data, it can be seen that the extremely limited water solubility of makosinone hydrochloride at pH 1.2 (corresponds to the pH value in the stomach on an empty stomach) sharply decreases in a buffer with pH 4.5 (corresponds to the conditions of the upper small intestine under fasting conditions) and practically loses solubility at pH 6.5, which corresponds to the middle section of the small intestine. It is this dependence that determines the presence of a narrow bioavailability window for a given active substance in the intestine, since already in the middle section of the duodenum, which most efficiently absorbs xenobiotics, pH rises to values of 6-6.5, at which macosinone loses its solubility.

In the same experiments, it was shown that the solubility of makosinone HCl does not change when the particle size in the substance powders changes in the range from 0.04 to 0.8 mm.

Example 2 The example describes the critical quality indicators set and monitored in the development and testing of gastro-retentive macosinone HCl tablets.

Example 3. A method for assessing the mucoadhesive properties of tablets by the force of separation of the tablet from the mucous membrane.

The equipment used is a texture analyzer (TA.XT plus, Stable Micro Systems, Great Britain).

Biomaterial - preparations of the pork stomach mucosa.

Each mucoadhesion test is performed on a freshly prepared porcine gastric mucosa preparation.

The tablet is attached to the flat end of a metal rod with a diameter of 10 ± 1 mm using a piece of double-sided adhesive tape.

Attach mucosal specimens to a slide using cyanoacrylate glue.

The probe of the texture analyzer with the tablet is lowered until it contacts the mucosa at a speed of 0.5 mm / s, the tablet is pressed against the mucosa with a force of 100 g for 300 s, and then returned to its original position at a speed of 0.5 mm / s. Tests are carried out on at least 3 samples at a temperature of 32 ± 2 ° C. The readings of the instrument, recorded automatically in the Exponent program, are used to construct the stress-strain curves.

With the help of the instrument software, the peel stress and adhesion work are calculated for the analysis of the obtained data.

The maximum stress σ _{max is} calculated by the formula:

σ _max = F _max / S,

where F _max is the maximum achieved value of the force at the stage of separation, S is the area of adhesive contact (the surface area of the tablet in contact with the mucous membrane). The work of destruction of the adhesive joint is calculated as:

where F is the current value of the force, l is the movement of the rod. Integration is carried out numerically, for example, by the trapezoidal method.

Example 4. A method for assessing bioadhesive (mucoadhesive) ability by the "wash-off" method.

The equipment used is a "paddle stirrer" dissolution tester (USP, Apparatus 2); 1000 ml glasses. Special holders for the stirrer blades.

The test medium is 0.01 N HCl.

Biomaterial - fragments of the mucous membrane of the pig's stomach.

Fragments of the mucous membrane are glued back to the holders using cyanoacrylate glue.

In a laboratory stand, a tablet is placed on a mucosal fragment moistened with water and pressed under a load of 10 g for 5 minutes. Attach the tablet holders to the stirrer blades, place it in the test medium in the beaker, and turn on the stirrer. The tests are carried out at a stirring speed of 25 rpm. The separation time of each tablet is recorded.

Example 5. Determination of the degree of release of macosinone from modified release tablets.

The test is carried out in a 500 ml vessel using a dissolution tester with a paddle stirrer. The dissolution medium is 0.01 N HCl.

Taking into account the saturation limit of the solution for an extremely poorly soluble substance, makosinone hydrochloride, the sampling mode is set in such a way that the concentration of the substance in the vessel during dissolution does not reach the saturating concentration (<100 μg / ml).

Sampling mode: after 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, etc., 300 ml of solution are taken from the glass and replaced with the same volume of solvent. In each sample taken, the concentration and amount of macosinone rrii is determined by the specified regulatory assay method (HPLC or other validated method).

To determine the total amount of the released substance for the entire sampling time, all calculated values of m _i are summed up:

where Ci is the concentration of the substance in each of the selected aliquots with a volume of 300 ml and Ck is the concentration of the substance in the last taken sample (for which the replenishment of the selected volume is not carried out).

The tablet according to the invention should provide a modified sustained release of makosinone in the gastric environment so that the concentration of makosinone in this environment does not reach the limit of its dissolution (80-100 μg / ml). The rate of release of macosinone from the tablet, calculated from the results of comparing in vitro and in vivo data on dissolution and pharmacokinetics of macosinone, should be in the range of 25-50 mg / h. FIG. 2 shows the target calculated cumulative curve of the expected release of macosinone over time in a dissolution medium of 0.01 n HCl from gastro-retentive tablets containing 500 mg of macosinone when tested in vitro (tests in apparatus of the "paddle stirrer" type, the volume of the dissolution medium is 500 ml).

Examples of pharmaceutical compositions (dosage forms) according to the invention

Example 6. Gastro-retentive tablet of composition GR 10 - a complex system using granules based on hydroxypropyl-beta cyclodextrin (inclusion complex) and an extra-granular matrix based on a mixture of carbopol + HEC.

Brief description of the composition and technology

The preparation of a gastro-retentive tablet (GR10) according to the invention based on a swelling and mucoadhesive matrix (Carbopol + HEC + CMC) using an inclusion complex based on cyclodextrin is carried out as follows.

A gastro-retentive tablet is prepared by a separate method in three main stages: 1) Obtaining a complex based on an active pharmaceutical ingredient (API) of macosinone hydrochloride and hydroxypropyl-beta-cyclodextrin in the form of "primary" granules;

2) Obtaining a "secondary" granulate using ready-made cyclodextrin granules coated with primary granules with a mixture of a binder with pronounced mucoadhesive properties (CMC) and low-polymer PEG, which acts as a solubilizer that promotes faster erosion of the tablet matrix in an aqueous medium and the release of cyclodextrin granules containing active substance;

3) Obtaining a tablet with the introduction into the intergranular space of the swelling polymer components forming the tablet matrix - carbopol and hydroxyethyl cellulose (which also have mucoadhesive properties at the same time).

Brief description of production technology

To obtain tablets, the technology of wet granulation is used.

• Preparation of raw materials

• Weighing of raw materials

Each type of raw material is weighed into separate clean containers.

Preparation of humidifier solutions.

To prepare cyclodextrin granules, purified water is used as a humectant solution (the mixture-humectant ratio is 2.5: 1).

Preparation of a mixture for tableting

Stage 1 Preparation of cyclodextrin granules (preparation of the inclusion complex of macosinone hydrochloride with hydroxypropyl-beta-cyclodextrin for use in gastro-retentive tablets according to the invention.

Preparation of dry mix

Macosinone hydrochloride (49 g) and Hydroxypropylbetadex (hydroxypropyl-beta-cyclodextrin) (25 g) are loaded into a granulator mixer and stirred at a rotation speed of 300 rpm until uniform (visual control).

Granulation

With constant stirring of the mixture, a humidifier solution (purified water) is introduced into the mixer-granulator at a mixture-humidifier ratio = 2.5: 1. Stir at a stirrer speed of 600 rpm for 3-5 minutes until granules are formed (visual control).

Drying of granules

The granules are dried in a drying oven. Residual moisture content of the mixture is 2.28%. Temperature when measuring humidity - 105 ° С.

Calibration of granules

The granules are sieved through a sieve with a mesh size of 0.5 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.5 mm.

Stage 2. Obtaining modified granules

Preparation of dry mix

The obtained cyclodextrin granules are introduced into the mixer-granulator, and sodium CMC, PEG is added. The components are mixed until homogeneous for about 1 minute (visual control).

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution is introduced (an aqueous solution of Polysorbate-80 in the ratio Polysorbate-80: purified water 1: 16.9). Stir for no more than 2-3 minutes until a fine friable granulate is formed (visual control).

Drying of granules

The granulate is dried in a drying oven. Residual moisture content of the mixture is not more than 1.2% (temperature when measuring moisture content - 105 ° C).

Calibration of granulate

The granulate is sieved through a sieve with a mesh size of 0.71 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.71 mm.

Stage 3. Obtaining a tablet by pressing granules into a matrix

The calibrated granulate, Carbopol and HEC are introduced into the mixer.

Stir until smooth for 5 minutes (visual control). Sifted lubricant sodium fumarate is loaded into the mixer and the components are mixed for 3 minutes until homogeneous (visual control). The tabletting mixture is discharged into a labeled container.

The tableting process is carried out on a tablet press using an oval biconvex press tool measuring 20.00 mm × 10.00 mm.

The tablets obtained after pressing are controlled according to technological parameters: hardness, abrasion, height, deviations from the average weight of 20 tablets (deviation from the average weight of individual tablets should not exceed 5%), as well as according to the characteristics established by the specification according to example 2 (table 2) on the gastro-retentive form of tablets.

A distinctive feature of the composition (tablet) according to the invention is the use for its creation of a separately prepared in the form of granules of a complex containing macosinone hydrochloride and beta-hydroxypropyl-cyclodextrin, capable of forming inclusion complexes with various molecules,

In the manufacture of cyclodextrin granules, the mass ratio of the substance of macosinone hydrochloride and cyclodextrin can vary (for example, 2: 1, 1: 1 or 1: 2), but with the preservation of the quantitative ratio of the mass of cyclodextrin granules and other components, which makes it possible, if necessary, to obtain tablets with different dosages according to the active substance (for example, 500, 250 or 125 mg).

A fundamentally important distinguishing point in the design of a gastro-retentive tablet according to the invention is the use of a combined base substance delivery system in which pre-prepared primary cyclodextrin granules (cyclodextrin complex of insoluble macosinone) are incorporated into secondary granules based on a mixture of a binder component with high bioadhesive properties (CMC) and readily water-soluble solubilizer (PEG). The combination of these two disintegrants of different types: CMC - a capillary-type disintegrant, which ensures the rapid penetration of water into the secondary granules and PEG - a swelling type disintegrant, accelerate the flow of water into the secondary granules, their destruction (erosion) and the release of a complex of the active substance with CD ...

At the third stage of the technology, to ensure long-term preservation of the shape and size of the tablet after entering the stomach, the obtained granules of complex composition after the stage of dry mixing are pressed together with a mixture of highly hygroscopic, swellable and at the same time bio-adhesive polymer components - insoluble in water. Carbopol is a polyacrylic cross-linked polymer of the swelling type with moderate bioadhesive properties and a highly bioadhesive gelling polymer, partially soluble in water, hydroxyethyl cellulose (which simultaneously exhibits the properties of a capillary disintegrant).

When it gets into an aqueous medium, such a complex tablet swells quickly due to the joint absorption of water, both by granules and by matrix components (Carbopol and HEC). Polymers passing into a gel form, primarily matrix components, give the swollen tablet a high mucoadhesive ability to attach to the gastric mucosa, and due to the formation of the surface layer of the gel, they also slow down the release of dissolved amounts of macosinone from the tablet, thereby preventing it from exceeding its saturating capacity in the stomach environment. concentration. Subsequently, water enters the deeper layers of the tablet, leading to the gradual destruction (with the participation of a solubilizer - water-soluble PEG) of the secondary granules, from which, in turn, the "primary" granules based on cyclodextrin included in the polymer matrix begin to be released, due to the increased the solubility of the active substance from which is provided not only an increase in the concentration of makosinone, but also an increase in its solubility in the upper intestine with a pH of 4.5-6.

Example 7. Gastro-retentive tablet of composition GR 18 is a combined system using mucoadhesive modified granules based on hydroxypropyl-beta cyclodextrin and an extra-granular matrix based on a combination of polymers PEO and HPMC.

Brief description of the composition and technology

Brief description of production technology

To obtain tablets, the technology of wet granulation is used.

• Preparation of raw materials

• Weighing of raw materials

Each type of raw material is weighed into separate clean containers.

Приготовление раствора увлажнителя

Preparing the humidifier solution

To prepare cyclodextrin granules, purified water is used as a humectant solution (the mixture-humectant ratio is 2.5: 1).

To prepare the granulate, a solution of Poly-sorbate-80: purified water in a ratio of 25: 1 is used as a humidifier.

• Preparation of a mixture for tableting

Stage 1. Preparation of cyclodextrin granules

Приготовление сухой смеси В миксер-гранулятор загружают макозинона гидрохлорид, гидроксипропил-бета-циклодекстрин. Проводят перемешивание компонентов до однородного состояния (контроль визуальный).

Preparation of dry mixture Macosinone hydrochloride, hydroxypropyl-beta-cyclodextrin are loaded into the mixer-granulator. The components are mixed until homogeneous (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution (purified water) is introduced. Stirring is carried out for 2-3 minutes until granules are formed (visual control).

Сушка гранул

Drying of granules

The granules are dried in an oven (fluid bed dryer). Residual moisture content of the mixture is 2.28%. Temperature when measuring humidity - 105 ° С.

Калибровка гранул

Calibration of granules

The granules are sieved through a sieve with a mesh size of 0.5 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.5 mm.

Stage 2. Obtaining modified granules

Приготовление сухой смеси В миксер-гранулятор вносят полученные циклодекстриновые гранулы, натрия КМЦ, ПЭГ. Проводят перемешивание компонентов до однородного состояния в течение 1 минуты (контроль визуальный).

Preparation of dry mixture The obtained cyclodextrin granules, sodium CMC, PEG are introduced into the mixer-granulator. The components are mixed until homogeneous for 1 minute (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution (an aqueous solution of Polysorbate-80) is introduced. Stirring is carried out for 2-3 minutes until a fine friable granulate is formed (visual control).

Сушка гранулята

Drying of granules

The granulate is dried in an oven (fluid bed dryer). Residual moisture content of the mixture is 1.83%. Temperature when measuring humidity - 105 ° С.

Калибровка гранулята

Calibration of granulate

The granulate is sieved through a sieve with a mesh size of 0.71 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.71 mm.

Stage 3. Obtaining a tablet by pressing granules into a matrix

Calibrated granulate, HPMC and PEO are introduced into the mixer. Stirring is carried out until uniform for 5 minutes (visual control).

Then, the sifted sodium stearyl fumarate is loaded into the mixer and the components are mixed for 3 minutes until homogeneous (visual control).

The tabletting mixture is discharged into a labeled container.

• Tabletting

The tableting process is carried out on a tablet press using an oval biconvex press tool measuring 20.00 mm × 10.00 mm. At the beginning of the process, the tablet press is adjusted for the production of tablets with an average weight of 1100.0 mg.

The tablets obtained after pressing are controlled according to technological parameters: hardness, abrasion, height, deviations from the average weight of 20 tablets (deviation from the average weight of individual tablets should not exceed 5%), as well as according to the characteristics established by the specification according to example 2 (table 2) on the gastro-retentive form of tablets.

This example describes a variant of the tablet according to the invention, obtained according to the same three-stage technology based on primary cyclodextrin granules with macosinone subjected to a second granulation stage followed by compression in a mixture with the components of the swellable-adhesive matrix, but in contrast to example 6 in this case as a highly swellable polymer polyethylene oxide, the degree of swelling of which, unlike Carbopol, does not change with increasing pH values, and another highly adhesive cellulose derivative, HPMC, is used for matrix components.

Since the mucoadhesive properties of PEO are low, but the degree of swelling is high, the tablets according to this example have a lower degree of mucoadhesion, but swell more intensively when placed in an aqueous medium.

Example 8. The example characterizes the effect of the formation of a complex of the substance of makosinone hydrochloride with hydroxypropyl-beta-cyclodextrin (according to examples 6 and 7) on the dissolution parameters of the active substance from the cyclodextrin granules obtained according to the invention (stage 1 of the manufacturing method described in examples 6 or 7).

Thus, in the complex with hydroxypropyl-beta-cyclodextrin, the solubility of makosinone significantly increases, with the most important increase (by more than 200%) in the solubility in the biorelevant FaSSIF medium, which simulates the composition of intestinal juice in the upper part of the duodenum (the main absorption zone active substance). It is also important that the solubility of the complex with cyclodextrin in the FeSSIF medium (simulator of intestinal juice in the middle section of the small intestine) also remains 2-3 times higher than the initial salt of the active substance. In fact, this provides an expansion of the relatively narrow "bioavailability window" of makosinone in the complex with cyclodextrin.

Obviously, this feature of the cyclodextrin complex makes a significant contribution to the achievement of the obtained technical result - a significant increase in the bioavailability of the active substance when using the gastro-retentive tablet according to the invention.

Examples of pharmaceutical compositions as comparative drugs Example 9. Preparation of a dosage form of immediate release macosinone - hard gelatin capsules with a dosage of 80 mg of macosinone hydrochloride (in terms of the free base), the reference drug in experiments on the study of pharmacokinetics in experiments on dogs.

Brief description of technology:

Preparation of raw materials

Each type of raw material is sieved separately: makosinone hydrochloride, through a sieve with a mesh size of 0.5 mm; crospovidone, povidone, lactose monohydrate, colloidal silicon dioxide through a sieve with a mesh size of 1.0 mm; magnesium stearate through a sieve with a mesh size of 0.32 mm. Sifted raw materials are collected in production containers, weighed on scales

Preparing the humidifier

Prepare a humidifier by mixing 0.69 kg of povidone and 3.9 kg of purified water until completely dissolved.

Obtaining a mass for encapsulation

Sifted macosinone hydrochloride (8.1 kg), crospovidone (0.7 kg), lactose monohydrate (16.6 kg) are introduced into the mixer-granulator and mixed for 10 minutes. slow stirring; after adding the first portion of the humidifier, the mixer stirrer is switched to the "fast revolutions" mode for 10-20 seconds, and then again in the slow stirring mode, the remainder of the humidifier is introduced and the mixture is stirred in the "fast revolutions" mode for 40-60 seconds. At the end of the process, the wet granulate is passed through an 8.0 mm sieve and dried in a fluidized bed dryer at a temperature of 40-55 ° C to a residual moisture content of no more than 4%.

The resulting granulate is additionally subjected to dry granulation on a calibrator with a hole diameter of 1.2 mm.

Dry granules are powdered with a mixture (0.7 kg of crospovidone, 0.83 kg of colloidal silicon dioxide, 0.14 kg of magnesium sterate)

Encapsulation

The finished mixture is filled into hard gelatin capsules (size No. 0), up to the mass of the capsule contents of 300 mg.

The study of the dissolution index of the dosage form of macosinone hydrochloride in the form of capsules according to example 9 (Fig. 3) showed that the solubility of this substance in the composition of the granulate filling the capsules slightly increases in comparison with the crystalline substance (see example 1), and in the dissolution medium 0 , 01 n HCl solubility limit is 70-80 μg / ml. The nature of the dependence of dissolution on the pH values of the medium also did not change: the solubility significantly decreased in a standard acetate buffer with a pH of 4.5 and turned out to be below 10 μg / ml in a phosphate buffer with a pH of 6.8. At the same time, as the pH increased, a tendency towards aggregation of particles and their partial precipitation was registered.

Example 10. Gastro-retentive tablet of composition GR 08, gastro-retentive swelling and bioadhesive (mucoadhesive) tablet with modified release of an active substance poorly soluble in water (reference drug analogous to the principle of gastro-retention).

This example describes a variant of a gastro-retentive tablet, which is based on an analogue known from the prior art, developed for a poorly soluble active substance with rapid (like macosinone) elimination from the body (acyclovir) and using the same delay principle as the tablet of the invention dosage form in the stomach - due to the combination in one tablet of the ability to significantly swell and the ability to mucoadhesion (Development and characterization of gastroretentive sustained-release formulation by combination of swelling and mucoadhesive approach: a mechanistic study / R. Sankar, Subheet Kumar Jain. // Drug Design, Development and Therapy 2013: 7 1455-1469).

The composition according to example 10 differs from the composition according to the invention (GR10 or GR18 according to examples 6 and 7) both in the design principle and in the manufacturing technology of the dosage form, which uses a "monogranular" (monolithic) system with the simultaneous inclusion of active pharmaceutical substances of makosinone hydrochloride and all other excipients, including polymers forming the basis of the matrix - Carbopol and polyethylene oxide.

Brief description of production technology

To obtain tablets, the technology of wet alcohol granulation is used.

• Preparation of raw materials

Взвешивание сырья

Weighing raw materials

Each type of raw material is weighed into separate clean containers.

Приготовление раствора увлажнителя

Preparing the humidifier solution

A 7% solution of Povidone K-30 in isopropyl alcohol is used as a humidifier solution.

• Preparation of a mixture for tableting

Приготовление сухой смеси

Preparation of dry mix

Macosinone hydrochloride, microcrystalline cellulose 101, Carbopol, PEO are loaded into the mixer-granulator. The components are mixed until homogeneous for 1 minute (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution is introduced. Stirring is carried out for 2-3 minutes until a fine friable granulate is formed (visual control).

Сушка гранулята

Drying of granules

The granulate is dried in an oven (or fluidized bed dryer). Residual moisture content of the mixture is 1.80%. Temperature when measuring humidity - 105 ° С.

Калибровка гранулята Гранулят просеивают через сито с размером ячеек 1,0 мм (0,710 мм). Остатки протирают вручную через сито с размером ячеек 0,710 мм.

Calibration of the granulate The granulate is sieved through a sieve with a mesh size of 1.0 mm (0.710 mm). The residues are rubbed by hand through a sieve with a mesh size of 0.710 mm.

Опудривание

Dusting

Calibrated granulate and sifted Aerosil are introduced into the mixer. Stirring is carried out until uniform for 5 minutes (visual control).

Sifted magnesium stearate is loaded into the mixer and the components are mixed for 3 minutes until homogeneous (visual control).

The tabletting mixture is discharged into a labeled container.

• Tabletting

The tableting process is carried out on a tablet press using an oval biconvex press tool measuring 20.00 mm × 10.00 mm. At the beginning of the process, the tablet press is adjusted for the production of tablets with an average weight of 1100.0 mg.

The tablets obtained after pressing are controlled according to technological parameters: hardness, abrasion, height, deviations from the average weight of 20 tablets (deviation from the average weight of individual tablets should not exceed 5%), as well as according to the characteristics established by the specification according to example 2 (table 2) on the gastro-retentive form of tablets.

Example 11. Tablet of composition GR 20: a variant of a gastro-retentive tablet, according to the design scheme "monogranular" (monolithic) system with granules based on CMC + MCC and a matrix based on HPMC (reference drug with a monolithic matrix used in a comparative experiment to study pharmacokinetics on dogs).

The main difference from the tablet according to the invention: the tablet is made, as in example 10, with a so-called "monolithic" matrix, all the components of which are included in the initially prepared granulate. The release of the active substance in this case occurs due to a combination of diffusion processes through the matrix material and release due to the gradual destruction of the matrix.

Brief description of the production technology.

To obtain tablets, the technology of wet granulation is used.

• Preparation of raw materials

Взвешивание сырья

Weighing raw materials

Each type of raw material is weighed into separate clean containers.

Приготовление раствора увлажнителя

Preparing the humidifier solution

Purified water with Polysorbate-80 is used as a humidifier solution (mixture: water ratio - 1.55: 1).

Приготовление сухой смеси• Preparation of a mixture for tableting

Preparation of dry mix

Macosinone hydrochloride, microcrystalline cellulose 101, Povidone K-90, HPMC and sodium CMC are loaded into the mixer-granulator. The components are mixed until homogeneous for 1 minute (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution is introduced. Stirring is carried out for 2-3 minutes until a fine friable granulate is formed (visual control).

Сушка гранулята

Drying of granules

The granulate is dried in an oven (fluid bed dryer). Residual moisture content of the mixture is -2.03%. Temperature when measuring humidity - 105 ° С.

Калибровка гранулята

Calibration of granulate

The granulate is sieved through a sieve with a mesh size of 0.710 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.710 mm.

Опудривание

Dusting

The mixer is loaded with the calibrated granulate and sieved sodium stearyl fumarate, and the components are mixed until homogeneous for 3 minutes (visual control).

The tabletting mixture is discharged into a labeled container.

• Tabletting

The tableting process is carried out on a tablet press using an oval biconvex press tool measuring 20.00 mm × 10.00 mm. At the beginning of the process, the tablet press is adjusted for the production of tablets with an average weight of 1100.0 mg.

The tablets obtained after pressing are controlled according to technological parameters: hardness, abrasion, height, deviations from the average weight of 20 tablets (deviation from the average weight of individual tablets should not exceed 5%), as well as according to the characteristics established by the specification according to example 2 (table 2) on the gastro-retentive form of tablets.

Example 12. Gastro-retentive tablet of the GR 12 composition: a variant of the swelling-adhesive matrix tablet of the modified release type of macosinone, similar to the tablet according to the invention (GR10 or GR18 according to examples 6 and 7) according to the design scheme using a combined (non-monolithic) system and a polymer matrix, but in which the primary granules are obtained without the use of cyclodextrin - based on cellulose derivatives (CMC + MCC). As an extragranular matrix in this tablet, a mixture of the same polymers was used as in the preparation of a tablet of composition GR08 (example 10) carbopol + PEO.

The tablet of this example is intended to evaluate the effect of cyclodextrin on the release (solubility) of an active substance.

The tablet of composition GR12 is made according to a three-stage scheme for constructing a gastro-retentive tablet similar to the variant according to the invention (examples 6 or 7), in which separately prepared granules containing the active substance are included in the composition of secondary granules with mucoadhesive and solubilizer, which in the third stage by dry mixing and the pressing is combined with the matrix-forming polymer components, which are distributed between the granules. The release of the active substance makosinone in this case, as in the case of the tablet according to the invention, occurs mainly through the initial erosion of the matrix (release of the granules) followed by dissolution of the substance from the released granules.

Brief description of the production technology.

To obtain tablets, the technology of wet granulation is used.

• Preparation of raw materials

Взвешивание сырья

Weighing raw materials

Each type of raw material is weighed into separate clean containers.

Приготовление раствора увлажнителя

Preparing the humidifier solution

For the preparation of granules, purified water is used as a humidifier solution (the mixture-humidifier ratio is 1.1: 1).

To prepare the granulate, a solution of Poly-sorbate-80: purified water in a ratio of 55: 1 is used as a humidifier.

• Preparation of a mixture for tableting

Stage 1. Preparation of granules

Приготовление сухой смеси

Preparation of dry mix

Macosinone hydrochloride, Crospovidone XL-10 is loaded into the mixer granulator. The components are mixed for 1 minute until homogeneous (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution (purified water) is introduced. Stirring is carried out for 2-3 minutes until granules are formed (visual control).

Сушка гранул

Drying of granules

The granules are dried in an oven (fluid bed dryer). Residual moisture content of the mixture is -2.79%. Temperature when measuring humidity - 105 ° С.

Калибровка гранул

Calibration of granules

The granules are sieved through a sieve with a mesh size of 0.5 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.5 mm.

Этап 2. Получение модифицированных гранул

Stage 2. Obtaining modified granules

Приготовление сухой смеси

Preparation of dry mix

The resulting granules, sodium CMC, PEG are introduced into the mixer-granulator. The components are mixed for 1 minute until homogeneous (visual control).

Гранулирование

Granulation

With constant stirring of the mixture in a mixer-granulator, a humidifier solution (Polysorbate-80 solution: purified water) is introduced. Stirring is carried out for 2-3 minutes until a fine friable granulate is formed (visual control).

Сушка гранулята

Drying of granules

The granulate is dried in an oven (fluid bed dryer). Residual moisture content of the mixture is 2.95%. Temperature when measuring humidity - 105 ° С.

Калибровка гранулята

Calibration of granulate

The granulate is sieved through a sieve with a mesh size of 0.71 mm. The residues are rubbed by hand through a sieve with a mesh size of 0.71 mm.

Stage 3. Obtaining tablets by pressing granules into a matrix

Calibrated granulate, Carbopol and PEO are introduced into the mixer. Stirring is carried out until uniform for 5 minutes (visual control).

Sifted sodium stearyl fumarate is loaded into the mixer and the components are mixed for 3 minutes until homogeneous (visual control). The tabletting mixture is discharged into a labeled container. • Tabletting

The tableting process is carried out on a tablet press using an oval biconvex press tool measuring 20.00 mm × 10.00 mm. At the beginning of the process, the tablet press is adjusted for the production of tablets with an average weight of 1100.0 mg.

The tablets obtained after pressing are controlled according to technological parameters: hardness, abrasion, height, deviations from the average weight of 20 tablets (deviation from the average weight of individual tablets should not exceed 5%), as well as according to the characteristics established by the specification according to example 2 (table 2) on the gastro-retentive form of tablets.

Example 13. Comparison of the properties of gastro-retentive tablets of different compositions.

A more clear comparison of all the above examples in terms of the composition of the compositions, as well as the controlled properties of the compositions, all the characteristics of the compositions described above are shown in Table 10. Based on the results of in vitro testing of gastro-retentive tablet variants selected at this stage of pharmaceutical development, tablet variants were selected for comparative tests in vivo to assess bioavailability and pharmacokinetics in large animals (dogs).

Thus, according to the results of pharmaceutical development and in vitro tests for compliance with the set values of the critical quality indicators, the tablets of the compositions GR08 (according to example 10) and GR12 (according to example 12) did not pass the tests for the parameters of swelling and mucoadhesion, respectively. The tablet of the GR12 variant (according to example 12) also did not meet the requirements for the rate of release of the active substance. At the same time, the tablet of the GR20 composition (example 11), despite the lower level of cumulative release of macosinone for 6 hours of dissolution (at the level of the minimum permissible established values (Table 2), had the same high rates and degree of swelling and mucoadhesion as and tablets of the invention (examples 6 and 7).

In this regard, for the stage of biological studies to assess the pharmacokinetics, tablets according to the invention of compositions GR10 (according to example 6) were selected, obtained according to the developed three-stage scheme using a complex combined matrix, and GR20 (according to example 11), as a comparison drug with no less, than the tablet according to the invention in terms of retention of the tablet in the stomach, but made according to an alternative scheme using a "monolithic" matrix and granules obtained without the use of cyclodextrin. Such a comparison drug allows experimentally in vivo experiments to evaluate the effect of the original method selected according to the invention for producing a gastro-retentive tablet based on the cyclodextrin complex of macosinone

In the same experiments, an immediate release dosage form (capsules) according to Example 9 was also used as a comparator.

Example 14. Study of pharmacokinetics.

The study of the pharmacokinetic characteristics of various dosage forms was carried out on dogs of the Beagle breed with a body weight of 10-12 kg, 5 animals per group. The maintenance and feeding regime of the animals was carried out in accordance with the regulatory documents.

The dosage forms were administered to the dogs as a single oral administration on an empty stomach.

Access to water was not restricted. The animals received access to food no earlier than 6 hours after the administration of the drugs. Blood for the study of the concentration of makosinone in plasma was collected according to the established schedule, starting from the control point 15 minutes, the last time point of sampling was 24 hours.

The content of makosinone in blood plasma was determined by a previously validated bioanalytical method using equipment for HPLC / MS analysis.

The curves of the pharmacokinetics of the tested formulations in comparison with the immediate release form (capsules) are shown in figure 4, the results of the calculation of the main pharmacokinetic parameters are shown in table. eleven.

As follows from the results presented in figure 4 and in table 11, in an in vivo study of the pharmacokinetics of the active substance makosinone in dogs after oral administration of various dosage forms, the results of evaluating the pharmacokinetics of the immediate release dosage form (capsules with a dosage of 80 mg) corresponded to those previously obtained in preclinical experiments on various animal species, as well as in clinical trials with the participation of volunteers, the data that with oral administration of this form there is a rapid increase in the concentration of macosinone in the blood (Tmax ~ 1 hour), followed by the same rapid decrease to practically undetectable concentrations after 8-10 hours. These results confirmed the conclusions drawn from the results of clinical studies that in order to achieve the necessary therapeutic effect on drug-resistant forms of the causative agent of tuberculosis, it will be necessary to apply a regimen with at least twice a day taking capsules with a total single dose of at least 640 mg; with a maximum permissible dosage of 80 mg / capsule, this will require taking at least 8 capsules of the drug at the same time twice a day. This dosing regimen, taking into account the complex nature of tuberculosis therapy, cannot be considered acceptable for clinical use.

In contrast to the immediate release form, both tested gastro-retentive tablets (variant according to the invention GR10 (example 6) and the comparison drug GR20 (example 11) differed significantly in pharmacokinetic parameters from the immediate-release drug (capsules according to example 9) in the form of the pharmacokinetic curve: after taking these tablets significantly (by 2-4 hours) shifts in time to the moment the maximum concentration of makosinone in the blood plasma is reached, the excretion of the drug is significantly slowed down.However, for the gastro-retentive comparison tablet of the GR20 composition (according to example 11), a significantly lower indicator of the maximum content of makosinone in the blood (C _max ) and, accordingly, the parameter of the area under the curve AUC and relative bioavailability (F _relative ).

It should be noted that in the course of in vivo tests carried out for a gastro-retentive tablet based on a cyclodextrin complex according to the invention (in particular, the GR10 composition according to example 6), the effect of the appearance of a significantly delayed in time (15-16 hours from the moment of administration) second peak of increase the concentration of the active substance in the blood plasma. Such a phenomenon was not registered for another gastro-retentive tablet tested in this experiment (composition GR20 according to example 11), obtained without using a cyclodextrin complex. At the same time, it should be taken into account that the GR20 tablet according to the results of in vitro tests was characterized by swelling and mucoadhesion parameters similar to the GR10 tablet, providing a long retention in the stomach, and in terms of the matrix erosion rate (recorded by the dissolution rate), the GR20 tablet was inferior to the composition in terms of the invention of GR10, which, in turn, reduces the likelihood of evacuation of this form from the stomach due to the rapid decrease in size compared to the tablet according to the invention. In addition, according to the results of this in vivo study, the gastro-retentive nature of the GR20 tablet is confirmed by a significant (more than 2 hours) shift in time of the peak of the maximum concentration, as well as a significant decrease in the rate of excretion of the active substance and an increase in the average residence time of the active substance in the body ( Table 11). The effect of the appearance of a second sufficiently long period of increase in the concentration of makosinone in the blood, registered in an experiment on dogs for tablets according to the invention (example 6) of composition GR10, of the composition GR10, 10-12 hours after administration of the tablet, is unexpected, since the the methods of design and methods of gastro-retentive tablets retention in the stomach, in particular, for swellable mucoadhesive tablets of acyclovir, the manufacturing method of which was taken as an analogue in the preparation of gastro-retentive tablets according to example 10, indicate that the retention time in the stomach of such dosage forms is usually no more than 4 -8 hours due to the gradual degradation of the tablet matrix and recurring at intervals of 2-4 hours the phenomena of periodic increased gastric motility ("migrating myoelectric complex"). A typical form in this sense for the case of testing gastro-retentive tablets was demonstrated by the composition comparison tablet GR20 according to example 11.

The two-phase pharmacokinetic curve described in the present invention, registered in experiments on large animals, for the variant of the swellable and mucoadhesive tablet according to the invention GR10 (example 6) with the appearance and such a prolonged shift of the second phase of the intake of the active substance into the body for such delivery systems is previously unknown from the prior art. It should be taken into account that the fundamental difference between the tablet according to the invention (example 6) and the comparison tablet (example 11) is the use of a cyclodextrin complex as part of a complex combined delivery system.

Thus, possessing the noted unusual properties, the tablet according to the invention (in particular, the composition GR10 according to example 6) provides an increase of at least 2 times (in terms of the total exposure to AUC) bioavailability and predicted therapeutic efficacy compared to the immediate release dosage form, which is caused not only by an increase (also by about 2 times) in the maximum concentration in blood plasma, but also by a significant (by about 3 times) increase in the residence time of the active substance in the body (MRT) (Table 11). It is important to emphasize that due to the unexpected, significantly delayed increase in the content of the active substance in the blood plasma, registered in these experiments, the concentration of makosinone in the blood plasma remains at a sufficiently high level for 22-24 hours after a single dose of the tablet.

From the prior art, specialists know that when predicting the therapeutic efficacy based on the results of pharmacokinetic studies, it is necessary to take into account not only the results of determining the concentration of the active substance in the blood, but also some additional factors associated with repeated administration of the drug during the course of treatment and the uneven distribution of the active substance in the body. Thus, according to the results of clinical studies, it has been shown that under conditions of repeated daily injections of makosinone, there is a significant, up to 50% increase in the maximum attainable concentration in blood plasma and an increase in the total retention time of the drug in the body. In addition, the results of preclinical studies on various animal species showed that makosinone, after entering the systemic circulation, is characterized by a pronounced delay in the lungs, spleen, and liver. The maximum concentration of the active substance in the tissues of these organs is 3-4 times higher than the concentration found in the blood plasma. Taking into account these data, the results of studies of the bactericidal activity of makosinone in vitro, as well as the presence of antimicrobial activity in some metabolites of makosinone, the target level of the therapeutically effective concentration of makosinone in human blood plasma can be set at 60-90 ng / ml. If we take into account the known value of the conversion metabolic coefficient for dogs (about 3), which was confirmed by the actual results of comparing the registered concentrations of makosinone in the blood of humans and animals at the same doses of the drug, then the indicator of the target minimum concentration in the blood plasma of dogs can be set at 20-25 ng / ml ... In the figure of Figure 4, this minimum threshold of effective concentration is marked with a horizontal line (7, 8). At the same time, inset B to this figure shows the predicted concentration of makosinone in the lung tissue, taking into account the coefficients of tissue bioavailability. From the data presented in figure 4, it follows that under the conditions of a single administration to animals of the tablet according to the invention, in particular, the composition GR10, the predicted content of the active substance in the main target organ will exceed the established threshold for at least 24 hours.

At the same time, comparison drugs - an immediate release form (capsules according to example 9) and a gastro-retentive tablet of composition GR20 according to example 11, obtained without using a cyclodextrin complex, do not have this ability, and already after 6-8 hours after a single dose, the predicted concentration of macosinone in the target organ (mild) is expected to be below the prescribed therapeutic level.

Based on the foregoing, it can be concluded that the gastro-retentive dosage form according to the invention at a dosage of 500 mg is capable of maintaining the concentration effective against M. tuberculosis bacteria and, accordingly, the effective therapeutic effect of macosinone on the cells of the tuberculosis pathogen in the human body under single dose of the drug.

At the same time, the dosage of a 500 mg tablet taken once a day is promising for use in clinical practice, including when compared with the dosage level (about 640-1000 mg at least 2 times a day) required to achieve in the clinic therapeutic effect when using forms of immediate release. This will significantly reduce the total drug load on the patient's body, which is especially important for complex multicomponent treatment regimens for tuberculosis infection.

Thus, the developed gastro-retentive tablet according to the invention and the method for its preparation provided a solution to the problem of the invention and the achievement of the claimed technical result.

Although the invention has been described with reference to the disclosed embodiments, it should be apparent to those skilled in the art that the specific experiments described in detail are provided for the purpose of illustrating the present invention only and should not be construed as in any way limiting the scope of the invention. It should be understood that various modifications are possible without departing from the spirit of the present invention.

Claims (26)

1. Gastro-retentive swellable mucoadhesive tablet for controlled release of macosinone in the stomach and upper part of the small intestine, which is a combined system comprising a complex of macosinone or a pharmaceutically acceptable salt thereof in a therapeutically effective amount and cyclodextrin in the form of granules, additionally coated with a layer of mucoadhesive polymer, including a solator and molded into a polymer matrix based on two hydrophilic soluble or hydrophilic soluble and insoluble swelling and mucoadhesive polymers, and at least one additional pharmaceutically acceptable excipient. 2. A tablet according to claim 1, wherein the pharmaceutically acceptable salt is hydrochloride, hydrogen sulfate, ethanesulfonate or methanesulfonate. 3. A tablet according to claim 1, wherein the cyclodextrin is 2-hydroxypropyl-beta-cyclodextrin, 6-monodeoxy-6-monoamine-beta-cyclodextrin, gamma-cyclodextrin, 2-hydroxypropyl-gamma-cyclodextrin, or sulfobutyl-beta-cyclodextrin ... 4. The tablet of claim 3, wherein the cyclodextrin is 2-hydroxypropyl-beta-cyclodextrin. 5. A tablet according to claim 1, wherein the mucoadhesive polymer coating the granules is a soluble carboxymethyl cellulose with a degree of substitution of at least 0.7. 6. A tablet according to claim 1, wherein the solubilizer is a medium molecular weight polyethylene glycol, a monoester of polyoxyethylated sorbitan and higher fatty acids. 7. A tablet according to claim 6, wherein the average molecular weight polyethylene glycol is PEG 6000 or PEG 8000. 8. The tablet of claim 6, wherein the polyoxyethylated sorbitan higher fatty acid monoester is Polysorbate 80. 9. A tablet according to claim 1, wherein the hydrophilic soluble mucoadhesive matrix polymer is a soluble cellulose derivative or high molecular weight polyethylene oxide with a molecular weight in the range of 1,000,000 to 4,000,000 Da. 10. A tablet according to claim 9, wherein the cellulose derivative is hydroxyethyl cellulose or hydroxypropyl methyl cellulose. 11. A tablet according to claim 1, wherein the insoluble mucoadhesive matrix polymer is a high polymer carbomer. 12. The tablet of claim 11, wherein the carbomer is carbopol. 13. The tablet of claim 1, wherein the pharmaceutically acceptable substance is a filler, binder, glidant, or lubricant. 14. A tablet according to claim 1, wherein the weight ratio of macosinone or a pharmaceutically acceptable salt thereof to cyclodextrin in the complex is from 0.5: 1 to 2: 1. 15. The tablet according to claim 1, in which the content of the complex of macosinone or its pharmaceutically acceptable salt with cyclodextrin is 50-80% of the mass. based on the total weight of the tablet. 16. A tablet according to claim 1, wherein the amount of macosinone is 125-500 mg. 17. A tablet according to claim 1, which has an original length of at least 20 mm and a width of at least 10 mm. 18. A tablet according to claim 1, the swelling index of which in an aqueous medium after 6 hours is 95-150%. 19. Tablet according to claim 1, the adhesion force of which in vitro is 0.01 N. solution of hydrochloric acid ensures its retention on the surface of the mucous membrane for at least 5 hours. 20. A tablet according to claim 1, characterized in that the rate of release of macosinone in vitro is 0.01 N. hydrochloric acid solution is 25-50 mg / h. 21. A tablet according to claim 1, characterized in that the release of the active substance in vitro in the dissolution medium is 0.01 N. HCl after 6 hours is 30-50%. 22. The use of a tablet according to claim 1 for the prevention and / or treatment of a disease caused by mycobacteria. 23. The use of claim 22, wherein the mycobacteria are M. tuberculosis . 24. Use according to claim 22, wherein the disease is tuberculosis. 25. The use according to claim 24, characterized in that tuberculosis is caused by strains of the pathogen that are sensitive and resistant to other drugs, including strains with multidrug or extensively drug resistance. 26. Use according to claim 22, wherein the tablet is administered orally once a day.

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