RU2457835C1 - Glycine-based pharmaceutical composition of prolonged action and method of its obtaining - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and represents pharmaceutical composition of prolonged action which includes non-agglomerated particles, containing active substance glycine and auxiliary substances, characterised by the fact that non-agglomerated particles represent nanoparticles with size from 170 to 500 nm, containing biodegradable polymer, as auxiliary substance polyvinyl alcohol as SAS, poloxamer as stabilising agent and cryoprotectant.

EFFECT: invention ensures increase of bioavailability of active substance glycine resulting from application in composition of biodegradable polymer, due to which coefficient of transmission and glycine assimilability increase, and time of glycine activity in organism grows.

4 cl, 5 ex

Description

The invention relates to the pharmaceutical industry, in particular to pharmaceutical compositions for the manufacture of a sustained release glycine preparation based on biodegradable polymers and excipients.

The pharmacological effect of glycine is reflected in a change in many functions, and the proposed spectrum of specific pharmacological activity and indications for its use are very diverse, although they are not always experimentally justified and do not always have clinical confirmation.

According to modern data, glycine is one of the most widely used drugs in neuropsychiatric clinical practice. It seems that its therapeutic effectiveness is due to:

- universal antistress effect, not accompanied by a sedative effect;

- the ability to normalize the state of the nervous system during the period of hyper-excitation, overwork and intoxication;

- normalization of the psychoemotional and neurological state;

- improvement of mental performance, memory;

- the presence of a nootropic effect;

- detoxifying effect of the drug in chemical (including medicinal) intoxications.

It should be noted other important properties of glycine, allowing it to be used for various diseases:

- the absence of contraindications and side effects;

- lack of age restrictions at admission;

- a wide range of selection of individual dosages;

- the ability to create integrated treatment regimens in combination with other drugs.

Glycine increases the effectiveness of both typical and atypical antipsychotic drugs. US Pat. No. 6,162,827 A1, "Treatment of negative and cognitive symptoms of schizophrenia with D-serine," patented a method for treating schizophrenia, namely, reducing the negative symptoms of schizophrenia, using glycine.

Glycine penetrates into most body fluids and body tissues, including in a limited number in the brain; it is metabolized to water and carbon dioxide, its accumulation in tissues does not occur.

A known prolonged-action pharmaceutical composition based on glycine, containing 98.1-99.8 wt.% Non-agglomerated particles of the drug substance - aminoacetic acid (glycine), coated with a polymer coating (microcapsules), including 97.6-99 wt.% Drug substance and 1-2.4 wt.% Film-forming, 0.1-1.0 wt.% Lubricating and 0.1-0.9 wt.% Water. A ripper, which is a medicinal substance, is introduced into the composition in an amount of 0.1-10 wt.% By weight of the mixture obtained. A method of obtaining a composition includes the preliminary preparation of crystals coated with a polymer shell (microcapsules) by applying a shell from a binder to non-agglomerated particles of a drug substance and adding a lubricating and disintegrating component. During the preparation of microcapsules and / or during their intermediate storage, the moisture content in the shell is adjusted to a concentration of not more than 0.5 wt.%, And before adding the lubricating and loosening components, the microcapsules are moistened with water to 0.1-0.9 wt.%. The resulting composition can be controlled by the time of disintegration without the use of additional excipients (RU 2171673 C1, publ. 10.08.2001). A drug based on such a composition is used sublingually. The disintegration time of the tablet is at least 10 minutes and not more than 30 minutes.

It is known that the presence of the blood-brain barrier (BBB) makes it difficult to treat many diseases of the central nervous system, since it does not miss a number of drugs, including glycine.

The poor permeability of glycine through the BBB encourages doctors to use multi-gram amounts of glycine (30-100 g per single dose), which, however, do not achieve a therapeutic effect, possibly due to the relatively rapid destruction without toxic effects, as well as due to insuperability of tissue barriers, particular BBB. Prolonged circulation in the bloodstream of optimal amounts of glycine can maximize the use of the low BBB bandwidth for glycine. The creation of delivery vehicles and the simultaneous deposition of glycine using nanoparticles increases both the bioavailability and effectiveness of glycine as a pharmacological agent.

The objective of the invention is the creation of a highly effective glycine composition of prolonged action, providing its high bioavailability.

Thus, the need to have a constant source of glycine, capable of maintaining the glycine level required for cells for a certain time, is the dominant pharmacological criterion when choosing the parameters of a new drug. It is proposed to use glycine-loaded nanoparticles as such a source or depot that maintains the optimal level of glycine in the current needs mode. In the nanosomal form, glycine is able to act both on the glycine reception fields in the local sublingual region, and in other glycine-prescription regions of the body, eliminating the physiological and even pathophysiological deficiency in this important mediator.

In connection with the foregoing, biodegradable polymers are best suited to create a glycine depot, which, when introduced into the body, are able to circulate for a long time in the body for a controlled time (hours-days).

The technical result of the invention is to increase the bioavailability of the active substance - glycine due to the use of a biodegradable polymer in the composition, due to which the transmission coefficient and digestibility of glycine increase and the time of glycine activity in the body increases.

The technical result is achieved by a pharmaceutical composition of prolonged action, including non-agglomerated particles containing drug glycine and excipients, in which non-agglomerated particles are nanoparticles ranging in size from 170 to 500 nm, containing a biodegradable polymer, polyvinyl alcohol (PVA) as a surfactant as a surfactant , poloxamer as a stabilizer and cryoprotectant in the following ratio of components, wt.%:

Glycine 1 ÷ 50 Biodegradable polymer 25 ÷ 65 Polyvinyl alcohol 15 ÷ 30 Poloxamer 0.3 ÷ 2.5 Cryoprotectant 5 ÷ 20

Moreover, as a biodegradable polymer, the composition includes a bioavailable substance selected from a number of polylactides, or polylactide / glycolides, or polybutyl cyanocrylate.

As a cryoprotectant, the composition includes D-Mannitol, or glucose, or starch in an amount of 5 ÷ 20 wt.%.

The technical result is also achieved by a method of obtaining the above-described pharmaceutical composition, including obtaining a mixture of a medicinal substance glycine with excipients and drying it to obtain non-agglomerated particles, in which a solution of a biodegradable polymer in an organic solvent (acetone, chloroform, etc.) is mixed to obtain a mixture of glycine with excipients. n.) with an aqueous solution of poloxamer and glycine, heat the mixture to remove organic solvent, filter it, d bavlyayut a cryoprotectant mixture and stirred, and drying the resulting mixture, it is performed by lyophilization to give a powder.

EFFECT: invention provides obtaining a new nanoform preparation of prolonged action "N-glycine" based on biodegradable polymers. The prepere consists of biodegradable polymer nanoparticles loaded with the active substance - glycine, which led to an increase in the bioavailability of the active substance, since along with the transport of free molecules, glycine associated with nanoparticles is expected to enter the bloodstream. The assimilation process will take place not only in the oral cavity, but also at different levels of the gastrointestinal tract (GIT), for this purpose a special polymer was used, namely biodegradable, i.e. enzymatically degradable. Thanks to the new composition, it becomes possible to reduce the frequency and dose of the drug. Also, the long-acting nanopreparation N-Glycine in the future will expand the range of indications for the use of the drug, in particular as an antipsychotic, and will increase the effectiveness of the pharmacotherapy course according to established indications.

The drug of prolonged action "Nano-Glycine", briefly "N-Glycine", which belongs to the metabolites, as a pharmacological agent, is referred to as the metabolite level of action drugs. The active substance of the N-Glycine sustained-release nanodrug is aminoxylot glycine. The drug "N-Glycine" in appearance is a powdery porous mass of white and yellow. It can be used for the preparation of powder, tablets, capsules, glycine solution, as well as an injection form of prolonged action.

The new drug in nanoscale dosage form has a sedative (sedative), mild tranquilizing (anti-anxiety) and weak antidepressant effect. According to the results of preclinical studies, the drug can be classified as practically non-toxic. Found: the dosage form is pyrogen-free and does not have an irritating effect.

The specific activity of "N-Glycine" was significantly higher than that of the comparison drugs (RU 2171673 C1), and appeared after 24 and 48 hours, which was not observed in the latter.

The composition and method for producing a powdered nanoscale dosage form of the drug "N-Glycine".

The proposed method is based on the emulsification method, which obtained nanoparticles based on a biodegradable polymer.

The composition of the proposed composition includes:

- glycine - the active substance.

- biodegradable polymer (polylactide, or polylactide / glycolide, or polybutyl cyanoacrylate, or other),

- polyvinyl alcohol (surfactant),

- poloxamer - stabilizer,

- cryoprotectant (D-Mannitol, or starch, or glucose, or other).

Polyvinyl alcohol is used as a surfactant - stabilizer of the secondary emulsion in the preparation of polymeric nanoparticles loaded with glycine.

Poloxamer is used as a stabilizer of the primary (water in oil) emulsion in the preparation of polymer nanoparticles. Poloxamers (other names - pluronics, proxanols) are nonionic surfactants, which are block copolymers of polyethylene glycol and polypropylene glycol with a molecular weight of 4-15 thousand. Poloxamers are used in the invention as a specialized means of a separate process step. The purpose of the stage is to obtain a relatively stable spatial distribution of glycine molecules, which is achieved by dissolving it in a poloxamer solution, which acts as a special reaction medium. This achieves a peculiar spatial “fixation” of glycine molecules. The creation of such a reaction medium, which is a dispersed system or a specific colloidal solution, opens up the possibility and, as we have established experimentally, becomes a necessary condition for the formation of nanoparticles loaded with glycine at subsequent technological stages, which can be converted into a finished dosage form.

The proposed composition may include various poloxamers, for example, Poloxamer-188 in an amount of 0.3 ÷ 1.5 wt.% Or Pluronic F127 in an amount of 0.5 ÷ 2.5 wt.%.

A cryoprotectant, for example, D-Mannitol, or starch, or glucose, is used to prevent the adhesion of nanoparticles during lyophilization, as well as to ensure the resuspension of the finished form to individual particles during the formation of a colloidal micellar system when used.

The following are examples of formulations of the proposed composition.

Example 1

Composition G1-1,

in wt.%:

Glycine 31.5 Polylactide / Glycolide PLGA 50/50 31.5 Polyvinyl alcohol 25.0 Pluronic F127 1,0 D-mannitol 11.0

The ratio of glycine: PLGA 50/50 is 1: 1.

Example 2

Composition Gl-2,

in wt.%:

Glycine 10.0 Poly (DL-lactide) -COOH 57.5 (PLA-COOH) Polyvinyl alcohol 24.5 Poloxamer 188 2.7 D-mannitol 5.3

The ratio of glycine: PLA-COOH is 1: 6

Example 3

Composition G1-5,

in wt.%:

Glycine 11.8 Poly (DL-lactide) 47.8 (DL-PLA) Polyvinyl alcohol 29.2 Poloxamer 188 fifteen D-mannitol 9.7

The ratio of glycine: DL-PLA is 1: 4.

Example 4

Composition Gl-4,

in wt.%:

Glycine 5.8 Polylactide / Glycolide (PLGA 50/50) 57.8 Polyvinyl alcohol 16,2 Poloxamer 188 0.6 Starch 19.6

The ratio of glycine: PLGA 50/50 is 1:10

Example 5

Composition G1-3,

in wt.%:

Glycine 19.7 RBCA polybutyl cyanoacrylate 39,4 Polyvinyl alcohol 28.1 Poloxamer 188 1.4 Glucose 11,4

The ratio of glycine: PBCA is 1: 2.

Table 1 shows the size distribution of licin-loaded nanoparticles of biodegradable polymers.

Table 1 Sample Codes Unimodal Analysis Particle size distribution Size value, nm The average value, nm Standard deviation, nm The coefficient of variation, % G1-1 590 502 21 4.2 G1-2 443 324 57 eighteen G1-3 352 279 75 27 Gl-4 256 168 90 53 G1-5 413 287 73 25 P≤0.05

Excess glycine can cause an uncomfortable state, but the drug is practically non-toxic according to the results of experimental preclinical studies.

It should also be noted that the dosage form of the drug “N-Glycine” has an irritating effect on the vein wall, integumentary tissues (skin, subcutaneous tissue), muscles and the pyrogen-free nature of the injected solutions. The new drug is well tolerated and harmless. The increased bioavailability of glycine allowed to reduce the intake of the drug by 5 times.

The proposed method for producing a pharmaceutical composition is as follows.

To obtain nanoparticles according to the developed technology, the active substance is added to the reaction mixture, which is a solution of selected biodegradable polymer

An example of obtaining N-glycine.

With stirring on a magnetic stirrer was dissolved in chloroform PLGA 50/50 and after 5 minutes received a solution of "A".

With stirring on a magnetic stirrer, glycine and poloxamer were dissolved in water and a solution “B” was obtained.

While stirring, solution “B” was added in portions to solution “A”. Homogenization was carried out to obtain a primary emulsion. PVA was added and rehomogenization was carried out with stirring.

Chloroform was evaporated under vacuum in a rotary evaporator. After evaporation, a glycine-polymer colloidal solution was obtained. This solution could contain large particles, for the separation of which they were filtered through a glass filter. D-Mannitol was then added and dissolved. Then, lyophilization was carried out for 20–22 h at 0.03–0.1 mbar. Received a white lyophilized powder.

Below are the results of animal tests in the test "Open field", confirming the high efficiency of the proposed pharmaceutical composition.

Open Field (OP) is a classic test technique for studying the behavior of rodents in unfamiliar stressful environments - an open, brightly lit area.

The study on rats was carried out on the installation - OP, which is a square platform measuring 100 × 100 mm, enclosed by a wall 400 mm high (material - matte polystyrene). The site is divided into 25 squares and has a uniform illumination of 90 lux.

The open field for mice is a circular arena with a diameter of 600 mm with a wall 600 mm high, divided into 36 equal sectors.

Table 2 shows the results of tests to study the effect of the claimed drug N-Glycine on rat behavior. A comparative analysis of the effect of N-Glycine and the reference drug - the substance Glycine on the behavior of white rats in the “open field” test 1 hour after the intravenous administration of drugs showed that N-Glycine has a pronounced significant anxiosedative effect in the entire range of tested doses (4, 25 mg / kg, 8.5 mg / kg, 12.75 mg / kg). In this case, the severity of anxiolytic and sedative components were significantly higher under the influence of N-Glycine, which indicates its higher efficiency.

So, the duration of the latent period of departure from the launch pad (LP) was 5-10 times higher in the N-Glycine groups than in the Glycine groups at the appropriate dosages. Moreover, the absolute value of the duration of the drug in the N-Glycine groups was less dependent on the dose of the drug (3.3 sec-5.0 sec) than in the comparison drug groups - the substance Glycine (0.3 sec-1.0 sec).

A comparative analysis of the effect of N-Glycine and comparison products Glycine produced by MHFP and Glycine produced by Biotika on the behavior of C55Bl / 6 - and BALB / c mice in the Open Field test after iv administration of the preparations (Table 3) showed that a single administration of N-Glycine had a significant calming effect over the entire range of tested doses (30 mg / kg, 15 mg / kg, 5 mg / kg), while iv administration of the comparison drugs Glycine (MCPP) and Glycine (" Biotics ") was accompanied by a significant difference from the behavior of animals of the control groups only under the action of drugs in doses of 30 mg / kg and 15 mg / kg and practically had no noticeable effect at a dose of 5 mg / kg. Moreover, the severity of the action of N-Glycine in the smallest dose (5 mg / kg) was equivalent to the action of the highest of the doses of the comparison drugs (30 mg / kg).

It is important to note that with the introduction of N-Glycine, the smallest of the doses tested (5 mg / kg) was equally effective as the largest of the doses tested with the comparison drugs (30 mg / kg). N-Glycine in the lowest dose tested had an effect on behavior indicators, and comparison drugs at this dose did not have a significant effect.

The result shows that N-Glycine produced a significant anxiolytic, mild sedative effect on the behavior of mice of inbred lines C55Bl / 6 and BALB / c when tested in the open field test.

A specific calming effect was more pronounced when tested on BALB / c mice, while N-Glycine had a more pronounced effect on the behavior of mice than the comparison drugs Glycine MCPP and Glycine Biotiki. An equally effective dose was 5-6 times lower than that of the comparison drugs (5 mg / kg with the action of N-Glycine relative to 30 mg / kg, with the action of the comparison drugs).

Tables 3-6 show the results of a comparative study of the delayed effects of the drug of prolonged action of N-Glycine in rats with different routes of administration.

Indicators of rat behavior in OP are prognostic characteristics of the level of anxiety (fear) and resistance to emotional stress. According to modern concepts, the behavior of an animal in an OP reflects mainly the realization of a conflict interaction of two motivations - an inherited program (instinct) for studying a new environment (reaction to novelty) and a self-preservation instinct reflected in an effort to minimize the possible danger from a new, unfamiliar environment (Leonard , 1989). The study of the new environment is carried out through the behavioral reactions of GDA and VDA. Racks are considered as a method of distant space inspection, as an indicator of research activity (Sarkisova et al., 1996), sensitive to the level of anxiety or the action of anxiolytics (Krupina et al., 1996). The tactics of inspecting the field in a direct way - exit to the center, waste towards the center, free intersection of space, is considered as an indicator of reducing the level of anxiety. In general, it is believed that a decrease in the overall mobility of animals in this test is a consequence of an increase in their level of emotional stress. The desire to minimize the possible danger from the new environment, in extreme manifestation, can cause complete immobility (reaction "freezing" (freezing)). Starting the animal directly on the floor of the arena creates conditions for recording the reaction of the animal to the emotional factor "surprise", while the placement of rats in the center of the arena in most cases initiates an avoidance reaction with a short latent period (LP), and the higher the level of anxiety, the shorter the duration LP. At a lower level of anxiety, animals start after an indicative reaction, which takes some time. The influence of classical tranquilizers is reflected in an increase in the value of the drug, the frequency and duration of exit to the central and adjacent sectors of the field.

In accordance with the above interpretation, under the influence of a single systemic administration of N-Glycine to rats with different routes of administration - both orally and intraperitoneally, a significant anxiolytic and calming effect of the drug on the behavior of rats when testing animals in the Open Field test after 1 hour was manifested 24 hours and after 48 hours from the time of a single administration of the drug, which was not observed in the experimental conditions after the administration of the substance Glycine or the comparison drug Glycine produced by MCPP.

N-Glycine 1 hour after administration (Table 2) had a pronounced significant anxiosedative effect in the entire range of tested doses (4.25 mg / kg, 8.5 mg / kg, 12.75 mg / kg), while the substance Glycine exerted significant changes in rat behavior only in large doses (8.5 mg / kg, 12.75 mg / kg). At the same time, the severity of anxiolytic and sedative components turned out to be significantly higher under the action of N-Glycine, which indicates its higher efficiency in relation to the comparison drug,

A single oral (iv) administration of N-Glycine to rats after 24 hours or 48 hours from the moment of administration (Table 6) had a significant anxiolytic and sedative effect over the entire range of tested doses (4.25 mg / kg, 12.75 mg / kg, 25.5 mg / kg), while the intravenous administration of the comparison drug Glycine MHFP or Glycine Biotiki was not accompanied after 24 hours (table 4) or 48 hours (table 5) from the moment of oral administration a significant change in behavior relative to indicators of control groups. Significant differences from the behavior indicators of animals in the control groups were not detected at any of the tested doses. Therefore, N-Glycine showed a delayed specific effect after 24 hours and after 48 hours, which indicates the prolongation of the specific anxiosedative effect of the drug. Comparison preparations do not possess this quality and type of activity.

In tests in mice, it was found that N-Glycine produced a significant anxiolytic and mild sedative effect on the behavior of mice of inbred lines C55Bl / 6 and BALB / c when tested in the Open Field test after iv administration (table 3). The specific calming effect of the drugs was more pronounced when tested on BALB / c mice, while N-Glycine had a significantly more pronounced effect on the behavior of mice than the comparison drugs Glycine MCPP and Glycine Biotiki. The equally effective dose of the N-Glycine preparation turned out to be 5-6 times lower than that of the comparison drugs (5 mg / kg with the action of N-Glycine relative to 30 mg / kg, with the action of the comparison drugs), which indicates a significantly greater effectiveness of the drug Glycine in this test.

The above results also confirm the higher efficiency of N-Glycine compared to the closest analogue, since glycine is present in it in the same form as in the comparison preparations used in the studies.

Claims (4)

1. A pharmaceutical composition of prolonged action, including non-agglomerated particles containing the active substance glycine and excipients, characterized in that the non-agglomerated particles are nanoparticles ranging in size from 170 to 500 nm, containing a biodegradable polymer, polyvinyl alcohol as a surfactant, poloxamer as a stabilizer and cryoprotectant in the following ratio of components, wt.%:
Glycine 1 ÷ 50 Biodegradable polymer 25 ÷ 65 Polyvinyl alcohol 15 ÷ 30 Poloxamer 0.3 ÷ 2.5 Cryoprotectant 5 ÷ 20 2. The composition according to claim 1, characterized in that as a biodegradable polymer nanosized carrier includes a bioavailable substance selected from a number of polylactides, or polylactide / glycolides, or polybulyl cyanoacrylate. 3. The composition according to claim 1, characterized in that as a cryoprotectant includes D-Mannitol, or glucose, or starch in an amount of 5 ÷ 20 wt.%. 4. A method of obtaining a pharmaceutical composition according to any one of claims 1 to 3, comprising obtaining a mixture of the active substance glycine with excipients and drying it to obtain non-agglomerated particles, characterized in that a solution of a biodegradable polymer in an organic solvent is mixed to obtain a mixture of glycine with excipients with an aqueous solution of poloxamer and glycine, heat the mixture to remove the organic solvent, filter it, add a cryoprotectant to the mixture and mix, and dry The mixture is obtained by lyophilization to obtain a powder.