RU2697411C2 - Composition for treating parkinson's disease - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.SUBSTANCE: present invention refers to pharmaceutical industry, namely to a pharmaceutical composition for treating Parkinson's disease. Pharmaceutical composition for treating Parkinson's disease in the form of nasal drops contains 3,4 dihydroxy-L-phenylalanine (L-DOPA), a biodegradable polylactide-glycolide milk/glycolic acid copolymer 50/50 (PLGA 50/50), cryoprotector D-mannitol, polyvinyl alcohol (PVA), olive oil, Tween 80 taken in certain amounts. Method for preparing the pharmaceutical composition involves preparing a solution of PVA of saturated L-DOPA; obtaining a suspension of L-DOPA in a solution of a copolymer of PLGA 50/50 in dichloromethane; obtaining an emulsion by adding a suspension of L-Dopa in a solution of a copolymer of PLGA 50/50 in dichloromethane to a solution of PVA, saturated with L-DOPA; second emulsification - homogenisation; obtained suspension composition L-DOPA with copolymer PLGA 50/50 is mixed with cryoprotector D-mannitol; olive oil is added to Tween 80; Then, lyophilised polymer particles of L-DOPA are added to the obtained mixture in PLGA 50/50 copolymer and mixed until homogeneous mass is formed under certain conditions. Use of the pharmaceutical composition for treating Parkinson's disease.EFFECT: disclosed pharmaceutical composition is bioavailable and effective for treating Parkinson's disease.3 cl, 9 tbl, 12 ex

Description

The invention relates to medicine, in particular to pharmacology and pharmaceutics, and relates to the nasal form of a new pharmaceutical composition in the form of drops containing 3.4 - dihydroxy - L - phenylalanine (levodopa) as an active ingredient distributed in a polymer matrix based on a biodegradable milk / glycol copolymer acids (polylactide glycolide, PLGA 50/50) for use in neurodegenerative diseases, in particular Parkinson's disease (PD).

PD is a chronic steadily progressing neurodegenerative disease with a wide range of motor and non-motor disorders. BP inevitably invalidates patients, significantly reducing the quality of life of patients and their families. After 10-20 years, 40-75% of patients die, and about 50% of survivors require constant care (Hely. M. A. The Sydney multicenter of Parkinson's disease: The inevitability of dementia at 20 years / MA Hely, WG J. Reid, MA Adena, GA Halliday, JGL Morris // Mov. Disord. 2008. - V. 23. - No. 6. - P. 837-844). The total prevalence of the disease in Europe among people over 65 is 1.6-1.8 cases per 100 people. Due to an aging population, Parkinsonism’s importance as a public health problem is expected to increase (Litvinenko I.V. Parkinson’s disease. M.: Miklosh, 2010. 216 p .; Handbook of Parkinson's disease / edited by Rajesh Pahwa, Kelly E. Lyons. - 4th ed. - 2007 .-- P. 501). The progression of the disease and the deterioration in the quality of life of patients, as well as insufficiently effective treatment create a serious social problem.

To date, levodopa remains the standard drug for PD. However, levodopa is characterized by a short half-life from blood plasma, which even in the best conditions of modern therapy leads only to pulsating stimulation of dopaminergic neurons. Therefore, long-term therapy is complicated by motor abnormalities and dyskinesia, which are a source of significant limitations for some patients. A therapeutic strategy that will ultimately deliver levodopa / dopamine to the brain in a more continuous and physiological manner will provide advantages over the standard form by reducing motor complications, which is essential for patients suffering from concomitant neurological or motor disorders (Olanow CW; Mov. Dis. 2008, 23 (Suppl. 3): S613-S622).

Currently, oral forms of levodopa have been developed with sustained release of the active substance. The use of liposomal dosage forms provides a prolonged action of drugs and a decrease in their toxicity. In During M.J. et al., it was shown that liposomes with DOPA incorporated, implanted in the partially denervated striatum of rats, showed a lasting effect for 40 days. Small unilamellar vesicles were obtained by scoring liposome dispersion. The results of these studies indicated a partial restoration of dopamine deficiency in animals with an apomorphine-induced BP model (During MJ, Freese A., Deutch AY, Kibat PG, Sabel B.A., Langer R., Roth RH Biochemical and behavioral recovery in a rodent model of Parkinson's disease following stereotactic implantation of dopamine-contatining liposomes // Exper. Neurol. 1992. V115. P. 193-199). Significant disadvantages of liposomal preparations include the high cost of the phospholipids used, as well as their low storage stability.

Attempts to continuously administer levodopa intraduodenally or by infusion were carried out using ambulatory pumps and patches. However, such treatments are considered extremely aggressive and uncomfortable, and can also cause dopaminergic side effects. Moreover, even continuous administration of levodopa or dopa-agonists is associated with periods of lack of action of self-limiting substances. (Nutt JG; Mov. Dis. 2008, 23 (Suppl. 3): S580-4).

The process of metabolic conversion of levodopa to dopamine is catalyzed by the aromatic L-amino acid decarboxylase enzyme, a ubiquitous enzyme present in the highest concentrations in the intestinal mucosa, liver, brain and capillaries of the brain. Due to the possible extracerebral metabolism of levodopa, it is necessary to administer large doses of this substance in order to obtain a high concentration of extracerebral dopamine, which in turn can cause nausea in some patients. Therefore, levodopa is usually administered in conjunction with the oral administration of a dopa decarboxylase inhibitor such as carbidopa or benserazide, which reduces the dose of levodopa necessary for a clinical response by 60-80% and thus prevents some of its side effects by inhibiting levodopa conversion to dopamine outside the brain. The process of reducing the dose of levodopa remains unstudied. Various formulations are well known containing levodopa alone or together with enzyme inhibitors associated with the metabolic destruction of levodopa, for example, decarboxylase inhibitors such as carbidopa and benserazide, catechol-O-methyl transferase inhibitors such as entacapone and tolcapone, and monoamine oxidase inhibitors (MAOs) -A or MAO-B, such as moclobemide, razagilin or selegiline, or safinamide. To date, the list of oral medications includes SINEMET® and SINEMET®CR sustained release tablets containing carbidopa or levodopa; STALEVO® tablets containing carbidopa, entacapone and levodopa; and MADOPAR® tablets containing levodopa and benserazide.

Thus, the problem of developing and compiling fundamentally new pharmaceutical compositions and methods for the delivery of the active substance, which are:

- provide long-term stimulation of L-dopa without the need to expand the composition of active substances;

- will make it possible to significantly reduce the dose of levodopa for a more effective treatment of motor disorders;

- reduce the likelihood of side effects.

This problem was solved by creating a nasal form in the form of drops containing the nanosomal form of the active substance 3.4 - dihydroxy - L - phenylalanine (levodopa).

Previously, a pharmaceutical composition was developed for the treatment of PD (RF Patent 2440100), containing food acid or a combination of food acids, providing a pH of the composition in the range 5.5-6.5, and a pharmacologically effective amount of micronized L-DOPA as an active ingredient, in which the active ingredient is present as particles on the surface of larger carrier particles. The effect of the drug is aimed at eliminating motor fluctuations in patients taking L-DOPA for the treatment of PD. The disadvantage of the composition is the relatively low effectiveness of the treatment. A drug has also been developed (RF Patent 2545734) for stopping the manifestations of PD based on micronized L-DOPA (3-hydroxy-L-tyrosine). However, to achieve optimal treatment results, such a system of delivery of the active substance is needed that could fully reveal the potential of the drug. It was precisely this technical result that came as a result of a series of experiments.

Note that the development of drug delivery systems is one of the rapidly developing and promising areas of pharmacology. Currently, various nano-sized carriers are successfully used, the indisputable advantage of which is the protection of healthy cells from the toxic effects of the drug and the increased bioavailability of innovative drugs. This reduces the likelihood of degradation of the included substance in the body, the effect of the drug substance is prolonged, its selective accumulation in the affected area is ensured. The drug substance, form and technology used to produce the drug are combined, so their interaction should be studied. For example, when choosing the materials used in the dosage form, it is important to understand the technological consequences of combining the selected elements. Similarly, knowledge of the drug release mechanism is key to understanding the critical characteristics of the drug and achieving the desired release profile. This knowledge can be used as an aid in the formation, design and development of new technology.

The essence of this invention is to develop such a form of pharmaceutical composition that will allow delivery of micronized 3.4 - dihydroxy - L - phenylalanine without loss of effectiveness and ensure a prolonged action. As a result of the experiments, it was revealed that this problem can be solved by preparing the pharmaceutical composition in the dosage form of nasal drops. The final result of the composition and method of producing the polymer composition is affected by the physicochemical properties of the drug substance itself, therefore, the optimal composition and preparation process were formed in the process of numerous experiments.

I. Pharmaceutical composition based on micronized 3.4 - dihydroxy - L - phenylalanine

When developing a nasal drug, the positive and negative aspects of existing known dosage forms used for both systemic and topical use were considered.

Intranasal cati. Among the advantages of this dosage form include the relative ease of manufacture and use, the disadvantages are the need to use cold storage conditions.

Intranasal aerosols (sprays) are one of the varieties of pharmaceutical aerosols. They have the following advantages: low consumption and uniform distribution of the active substance on the surface of the mucosa; possibility of application in various conditions; high concentration of the substance at the site of the pathological process. However, there are a number of significant drawbacks: propellants (propellants) can be irritating to the nasal mucosa; Mandatory synchronization of the drug administration with the moment of inspiration is required, which is difficult to achieve in children, elderly patients and patients with a low level of intelligence; with inept use, there is the possibility of damage to the nasal mucosa by the nozzle of an aerosol can; when spraying, the possibility of substances in the eyes, on the skin of the face, etc. is not excluded

Nasal gels. Gel is a mild topical dosage form. Advantages of the dosage form: prolonged action; possibility of use at night; the presence of a moisturizing effect on the nasal mucosa; a beneficial effect on the nasal mucosa with its dryness, the presence of crusts; upon contact with skin or clothing, the gel is easily washed off with water, leaving no residue (unlike ointments). Disadvantages: not all active substances can be introduced into the composition of the gels and, accordingly, be used in this dosage form; instability of the dosage form; the possibility of delamination during storage at the IUD and the aqueous phase; diffusion of the active substance into the tissue from the gel dosage form is slower than from the solution; the difficulty of applying with an abundant amount of mucous discharge.

Nasal ointments. Ointments for the nose are prepared under aseptic conditions. Since they are applied to wet mucous membranes, surfactants are added to increase their absorption. Advantages, ointment base provides a longer action of the active substances on the nasal mucosa; has a softening effect on the nasal mucosa; the dosage form of the ointment has a systemic effect and makes it possible to co-administer the active substances of both hydrophobic and hydrophilic nature into one drug.

Given the dispersion of particles of the obtained substance (levodopa in polylactide glycolides), the need for systemic exposure, the relative duration of stay on the nasal mucosa to ensure complete absorption, the regularity of the drug application on the nasal mucosa, as well as its use by elderly people, drops and gels were chosen as dosage forms ointments. Thus, the choice of excipients in the development of the dosage form of the drug for nasal use will be determined by the following factors:

- the properties of the administered pharmacologically active component;

- physico-chemical parameters of the nasal mucosa and nasal secretion;

- physico-chemical and technological properties of the excipients themselves.

II. The composition of the pharmaceutical composition based on micronized 3.4 - dihydroxy - L - phenylalanine

In the process of selecting the optimal form and composition, several compositions were developed for each of the selected finished dosage forms. As examples below, more preferred formulations will be given. The remaining compositions not indicated in the examples will be indicated in the description of the manufacturing technology.

Example 1. Water purified as a dispersion medium

Sample K-3 aq.

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients.

benzyl alcohol - 0.10;

sorbitol 70% - 0.02;

hypromellose - 0.05-0.50;

purified water - up to 10.0

Example 2. Vegetable oils as a dispersion medium. Sample K-3 oils.

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients.

tween-80 - 0.01;

olive oil - up to 10.0

Example 3. Combined systems as a dispersion medium

Sample K-1 comb.

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients:

olive oil - 0.5-2.5;

tween-80 - 0.005-0.02;

propylene glycol - 0.05-0.5;

sorbitol 70% - 0.02;

purified water - 2.20-4.43

Example 4. Combined systems as a dispersion medium. Sample K-2 comb.

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients:

olive oil - 2.0;

tween-80 - 0.02;

propylene glycol - 0.1;

sorbitol 70% - 0.02;

purified water - 2.86

Example 5. Gel Sample G-1 gel

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients:

sodium hydrogen phosphate anhydrous - 0.01;

citric acid monohydrate - 0.02;

sorbitol 70% - 0.1;

olive oil - 2.0;

tween-80 - 0.01;

hypromellose - 0.09-0.14;

purified water - 2.72-2.77

Example 6. Ointment Sample M-3 ointment

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0;

Excipients:

petroleum jelly - 3.0;

liquid paraffin - 1.5;

softizan 649 - 0.5

Example 7. Ointment Sample M-1 ointment

Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 - 5.0

Excipients: Vaseline - 5.0

The manufacturing technology of aqueous nasal drops

Sample K-1 aq. prepared as follows (per 100 ml). Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in 50/50 polylactide glycolide in an amount of 50.0 were weighed on a balance with an accuracy of 0.01 g, purified water was added to 100 ml. Shaken until a homogeneous mixture. A colloidal solution was formed, stable for 15-20 minutes. Then a partial separation of the system took place, and part of the particles settled to the bottom of the vessel. In assessing the stability of this sample and its suitability for use, it was concluded that it is necessary to introduce thickeners in the composition of drops, such as cellulose derivatives, sorbitol.

Samples K-2 aq., K-3 aq. and K-4 aq. prepared also per 100 ml. Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in 50/50 polylactide glycolide in an amount of 50.0 were weighed on a balance with an accuracy of 0.01 g. Solutions of hypromellose and methylhydroxypropyl cellulose of various concentrations from 0.5% were preliminarily prepared. up to 5%, with a concentration step of 0.5%. Sorbitol was used as a 70% aqueous solution. In the part of purified water, depending on the recipe, the calculated amount of disodium edetate, sodium citrate, sodium dihydrogen phosphate dihydrate, disodium hydrogen phosphate dihydrate was dissolved. The remaining water was added to the polymer particles suspended in the stand, shaken, cellulose solutions were added, and salt solutions were added last.

When using 2-5% solutions of hypromellose and methylhydroxypropyl cellulose, a stable suspension (suspension) was obtained that did not delaminate at a temperature of 4 ° C (storage temperature of polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50) within 14 days (observation period)).

The technology of manufacturing nasal drops with vegetable oils

Sample K-1 oil. and sample K-2 oil. prepared as follows (per 100 ml). Polymer particles with 3,4-dihydroxy-E-phenylalanine (L-DOPA) in 50/50 polylactide glycolide in an amount of 50.0 were weighed on a balance with an accuracy of 0.01 g, olive oil / sunflower oil in an amount of 50.0 g was added. until a homogeneous mixture is obtained. A suspension was formed which was stable for 60-90 minutes. Then a partial separation of the system took place, part of the particles settled to the bottom of the vessel. When assessing the stability of this sample and its suitability for use, it was concluded that it is necessary to introduce surfactants into the composition of drops, such as tween-80.

Samples K-3 oil. and K-4 oil. prepared in a similar way with the introduction of Tween-80. This made it possible to increase the sedimentation time of particles to 10 days at a temperature of + 4 ° C (observation time).

When choosing systems, compositions with a varying degree of hydrophilicity were tested, which ensured the introduction of propylene glycol or cetyl alcohol and liquid paraffin.

To clarify the effect of excipients on the sedimentation stability of the obtained combined drops and to identify the nature of their functioning on the model mucosa, the composition was made in the variants indicated in examples 3 and 4, as well as in the description of the manufacturing technology of combined nasal drops.

The technology of manufacturing combined nasal drops

Sample K-1 comb. prepared as follows (per 100 ml). Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 in an amount of 50.0 were weighed on a balance with an accuracy of 0.01 g, placed in a support, tween-80 was added, shaken, for uniform distribution of tween-80 polymer particles. Olive oil was added in the calculated amount and again shaken, a concentrated suspension (paste) was formed. Propylene glycol, sorbitol, and the calculated amount of water were placed in a mortar. Triturated to obtain a homogeneous mixture. The resulting liquid was added to the oil paste in parts, with constant shaking. A suspension emulsion system was formed, observed also under a microscope. It should be noted that the stability of the resulting system largely depends on the concentration of excipients.

The greatest stability was achieved with the ratio of ingredients specified in Example 4.

Samples K-2 comb. also prepared per 100 ml, however, given the significant volume of the lipophilic phase, the lipophilic part of the ointment was prepared separately. Cetyl alcohol and weighed amounts of olive oil and liquid paraffin (petroleum jelly) were placed in a mortar heated to 50–60 ° C; the mixture was thoroughly triturated until a homogeneous mass was formed. Polylactide particles moistened with tween-80 were added to the resulting mixture, mixed, and purified water was gradually added. All samples obtained were characterized by sedimentation stability for 10 days (observation period).

Nasal gel manufacturing technology

Sample K-1 gel was prepared as follows (per 100 ml). Polymer particles with 3,4-dihydroxy-L-phenylalanine (L-DOPA) in 50/50 polylactide glycolide in an amount of 50.0 were weighed on a balance with an accuracy of 0.01 g, placed in a mortar, tween-80 was added, gently mixed with a scraper, preventing grinding particles. Olive oil was added in the calculated amount and again mixed with a scraper. A concentrated suspension (paste) was formed. A 3-5% solution of hypromellose was placed in another mortar, into which the following sequence was introduced: sodium hydrogen phosphate anhydrous, sorbitol, citric acid monohydrate. Triturated to obtain a homogeneous mixture. The resulting mass was added to the oil paste in parts with constant stirring. A stable system was formed, the heterogeneity of which could be observed under a microscope.

Currently, nasal gels are represented on the pharmaceutical market of Russia by such drugs as Galazolin, Vibrocil. No systemic drugs were identified for nasal use. Perhaps this is due to the accuracy of dosing of nasal gels, which are packaged in tubes with an elongated tip and dosed by direct extrusion from the tube. For systemic nasal gels, this method is not suitable, since it requires dosing on a graduated spatula or other device, and only then introducing it into the nasal cavity onto the mucosa.

In the patent literature, insufficient attention has been paid to the topic of nasal forms. All the above facts were taken into account when developing formulations based on hypromellose in a concentration of 3-5%.

The technology of manufacturing nasal ointment

In the simplest case, the ointment composition may be represented by a suspension ointment with polymer particles on white paraffin (petroleum jelly) in accordance with the composition specified in Example 7.

A pre-weighed petroleum jelly was placed in a mortar heated to a temperature of 50-60 ° C, melted and polymer particles were gradually introduced with constant stirring.

The ointment obtained can be characterized as a fairly concentrated mass, therefore, softhisan 649 was introduced into the next sample along with paraffin oil. A pre-weighed petroleum jelly was placed in a mortar heated to a temperature of 50-60 ° C, melted, vaseline oil was added, softisan 649 was mixed. With constant stirring, polymer particles were gradually introduced. The obtained ointment was characterized by satisfactory characteristics, ductility, did not exfoliate, had visual uniformity.

In relation to nasal ointments of systemic preparation, it is also advisable to use a graduated spatula when dosing.

When developing a nasal ointment, given the duration of its use, macrogol of various brands were not used because of their osmotic activity and, accordingly, the potential for negative effects on the mucosa. The use of alginates (the formation of compressible films) was also considered inappropriate.

III. The choice of the optimal composition of the pharmaceutical composition based on micronized 3.4 - dihydroxy - L - phenylalanine

The choice of the optimal composition of nasal compositions was carried out using the developed model system that simulates the nasal mucosa.

In order to determine the most rational compositions of the developed nasal compositions with polymer particles of 3,4-dihydroxy-L-phenylalanine (L-DOPA) in polylactide glycolide 50/50 for the treatment of PD, their behavior was evaluated on an in vitro model system. The assessment was carried out by indicators of the distribution area and the degree of adhesion on the model surface. The study of the adhesive properties of the developed drugs was evaluated by the rate of runoff of the sample on the inclined surface of the model substrate moistened with a model medium.

For a more accurate determination of droplets, depending on the composition, 0.001% methylene blue (water-based drops) or a Sudan III drop was introduced.

The test agent in an amount of 5.0 ml, which amounted to levodopa in terms of the dose of the drug administered, was applied to the start line of a substrate preheated to 37 ° C (surface in a Petri dish). The substrate was raised at angles of 10 ° and 70 °, and after 30 seconds, the distance traveled by the front boundary of the liquid drop was measured. Runoff speed v (mm / s) was calculated by the formula:

where: L is the path traveled by a drop of liquid, mm;

τ is the runoff time, s.

The results of the experiment are presented in Table 1.

To measure the distribution area, the test agent colored in the amount of 5.0 ml, which was calculated as levodopa, was used to calculate the dose of the drug administered, placed on the surface of the model mucosa pre-heated to 37 ° C, located in a Petri dish, covered with a lid and thermostated at at a given temperature for 15 minutes (the time it may be on the mucosa), noting the distribution area for each sample. The distribution area S (cm ² ) was determined using graph paper.

The results of the experiment are presented in Table 2.

Analyzing the results of the experiment, it can be noted that water-based nasal drops without inclusion of cellulose derivatives are distributed over the model surface and, having low adhesive properties, cannot ensure dosing accuracy due to leakage from the nasal cavities. The introduction of hypromellose and methylhydroxypropyl cellulose can reduce the distribution area on the surface of the model mucosa. Oil-based nasal drops were distributed (Sample K-1 oil) extremely unevenly.

The developed nasal drugs can be arranged according to the behavior on the model mucosa (according to the suitability for further studies) as follows: ointments → gels → drops on a combined basis → drops on an oil basis → drops based on aqueous solutions of cellulose derivatives → drops based on aqueous solutions. Given the results of the experiments, the following samples were selected for further research:

Sample K-3 aq. - 1% hypromellose

Sample K-4 aq. - 1% methylhydroxypropyl cellulose

Sample K-2 oil.

Sample K-3 oil.

Sample K-4 oil.

Sample K-1 comb.

Sample 2 comb. one

Sample 2 comb. 2

Sample 2 comb. 3

Sample 2 comb. four

Sample 2 comb. five

Sample G-1 gel 3% hypromellose

Sample G-1 gel 4% hypromellose

Sample G-1 gel 5% hypromellose

Unexpectedly, in the process of work, it was found that reducing the dose of the auxiliary substance and adding it together with tween-80 at the stage of preparing the finished composition gave the most effective result in terms of adhesive properties, distribution area and speed of action, as well as prolongation of the active substance, taking into account all chemical physical properties of the micronized form of 3,4-dihydroxy-L-phenylalanine.

A preferred sample of the composition is shown in Table 3. The method for its preparation and effectiveness are recorded below.

IV. Pharmacological effectiveness of the optimal composition and comparative assessment of the specific activity of an innovative drug based on 3,4-dihydroxy-1-phenylalanine (levodopa) and polylactide glycolides in the BP model

The specific activity of an innovative drug with the optimal composition shown in Table 3 and the standard drug Levodopa with an antiparkinsonian effect were evaluated using the developed model of PD and a battery of behavioral tests.

According to current recommendations, a preclinical study of the activity of potential anti-Parkinsonian drugs is carried out on models of Parkinson's syndrome. In pharmacological modeling, PD usually resort to either various methods of inhibiting dopaminergic (DA) transmission, primarily in the nigrostriatum, or to activating cholinergic (CE) neurons. Two types of these models differ in the dynamics of the development of the process, as well as its manifestations, and are considered as reproducing, respectively, the akinetic-rigid or trembling forms of PS.

Since the developed drug based on polymer-encapsulated 3,4-dihydroxy-L-phenylalanine (levodopa) in polylactide glycolides (PLGA) is focused on the prolonged release of the active substance, the model with inhibition of DA transmission is more consistent with the necessary dynamics of the gradual development and duration of conservation of the simulated PD .

Model selection

In order to adequately formulate PD and its subsequent treatment with a potential anti-Parkinsonian drug, the pharmacological model of PD was used, which was induced by the destruction of the nigrostriatal dopaminergic system by neurotoxin 6-hydroxydopamine (6-OHDA) introduced into the rat brain.

The 6-OHDA model is considered a rat PS model, as they are most susceptible to neurotoxin, however, the manifestations of its effects in rats and monkeys have also been studied. 6-OHDA causes a characteristic change in animal posture - turning the head and tail deviation towards the injured side, as well as typical circular movements and stereotypes (sniffing, licking, biting).

Detailed preparation of a pharmaceutical composition

At the first stage, a solution of PVA saturated with L-DOPA is obtained. L-DOPA is placed in a glass vessel, a 2% PVA solution is added. Then the vessel is transferred to a water bath located on a magnetic stirrer, where L-DOPA is dissolved for two hours with constant stirring at a speed of 400-500 rpm. The temperature in the vessel should not exceed 37-42 ° C. Mixing the mixture in the bath lasts 30-40 minutes (without additional heating). Then the bath with water is removed, mixing continues for another 30-40 minutes until the solid phase is completely dissolved.

Then, a suspension of L-DOPA in a solution of PLGA 50/50 polymer in dichloromethane is obtained. In a glass bottle, L-DOPA is weighed. In a glass beaker, the PLGA 50/50 polymer is weighed and transferred to the reactor. Dichloromethane is weighed in a plastic beaker. Dichloromethane was transferred to the reactor portionwise and closed with a foil lid. Dissolution of the polymer is carried out with stirring with a paddle mixer of a mixing device at a speed of 300-400 rpm for 20-25 minutes

After complete dissolution of the PLGA 50/50 polymer in dichloromethane, the previously weighed portion of L-DOPA substance is loaded into the reactor, closed with a lid, mixing is continued at a speed of 300-400 rpm for 25-30 minutes until a homogeneous mixture is obtained. The resulting suspension solution is used to create an emulsion.

A 2% solution of PVA saturated with L-DOPA was transferred to a plastic reactor and placed in a water bath. The temperature of the reaction mixture should be maintained between 21-27 ° C. With vigorous stirring (600-700 rpm), the previously obtained L-DOPA solution-suspension with a pipette dispenser in 50-75 ml portions is gradually added to the central part of the reactor. The plastic reactor was covered with a foil lid; stirring was continued for 25-30 minutes. Stop heating and drain the water from the bath. The space of the bath tank, free from the reactor, is filled with ice.

Next, the mixture is subjected to a second emulsification (homogenization). To do this, a stand with a homogenizer is placed near the reactor and the N-27 nozzle (code S25N-25F) is immersed to the bottom. The mixture is emulsified at 24 thousand rpm and room temperature 3 times for 30-40 s, with two interruptions of 0.5 minutes. During the process, the formation of foam mass.

Next, the emulsion obtained is further processed using a submersible probe of a homogenizer (B. Braun, USA) GV-28, ultrasonic dispersion is carried out in the following mode: 1 min; 1.5 min; 1 min with two breaks of 1 min each (power level 300-320, repeating duty cycle 0.7 s). Dichloromethane was evaporated from the resulting product.

The homogenizate from the reactor is transferred to an E-29 flask with a capacity of 10 liters. The flask is filled about 1/3 of the volume, then connect it to the shafts of the rotary evaporator through a glass drop trap, connect the vacuum from the membrane (or water-jet) vacuum pump. After a set of vacuum in the system (approximately 0.4 units) include the rotation of the shaft. The mixture is evaporated on a rotary evaporator (evacuation using a water-jet pump) in the mode: 30-35 min at room temperature; 15-20 min at 25 ° C (in the bath); 10 min at 35 ° C; 10 min at 40 ° C. At the beginning of the evaporation of dichloromethane (at room temperature and 25 ° C), strong foaming and partial penetration of the foam into the trap and the receiving flask take place, therefore this stage of the process should be visually monitored and, if necessary, let air into the system through a special defoaming cock. The remaining amount of the mixture is gradually added. The process is completed when a vacuum of 0.93-0.98 units. and the effect of fogging traps.

After evaporation of the dichloromethane, a suspension of the composition of L-DOPA and PLGA 50/50 polymer in the flask was transferred to the mixing step with D-Mannitol. First of all, D-Mannitol 41.00 ± 0.01 g is dissolved in purified water 287.00 ± 1.00 ml. The addition of water to a sample of D-Mannitol is carried out at least 1 hour before the start of the mixing operation with the previously obtained suspension due to its slow dissolution.

Next is the step of filtering the suspension and mixing with D-Mannitol. The D-Mannitol solution was transferred to a Bunsen flask. The previously obtained suspension in the flask may contain large particles, for the separation of which the filtering process is carried out through a glass filter with a pore size of 40-100 μm (pore 111). The suspension is filtered by gravity, if necessary, the system is connected to a vacuum line. The combined filtrate was collected in a collection flask. After filtration is complete, the flask is disconnected and transferred to a magnetic stirrer.

The solution of D-Mannitol, the filtrate of the suspension and the washings are mixed in a receiving flask using a magnet in a fluoroplastic shell and a magnetic stirrer for 10-15 minutes at 300-400 rpm. Then determine the mass of the mixture in the flask. After that, the filtrate is prepared for freeze drying.

The mixture of the filtrate with D-Mannitol obtained at the previous stage is uniformly dosed into plastic jars (containers). The process is carried out in a room with a laminated air flow. Filled jars are covered with screw caps and placed in the freezer for freezing at a temperature of minus 70 ± 2 ° C. The freezing time is at least 24 hours. Next is the lyophilization process. The drying process is carried out at room temperature in the room for 45-48 hours. After this stage, the process of obtaining nasal drops is started.

Olive oil is poured into a sample of Tween-80 in a glass, mixed until the mixture is homogeneous. Lyophilized polymer particles with 3,4-dihydroxy-L-phenylalanine in 50/50 polylactide glycolide are transferred to the bowl. A mixture of Tween-80 and olive oil is transferred from a glass, distributing over the entire surface of lyophilized particles. The mixer is turned on at a speed of 50-60 rpm. Mixing is carried out for 5-10 minutes, achieving uniform distribution of the oil mixture in the entire volume of lyophilized particles. The parts add the remaining olive oil, mix thoroughly until a homogeneous mass is formed.

Finished nasal drops are a new suspension of Levodopa (polymer particles with 3,4-dihydroxy-G-phenylalanine in polylactide glycolide 50/50 1: 9) in a solution of tween-80 in oil. Weigh on the scales with an accuracy of 0.01 g to 10.0 g in glass bottles and seal with a cap with a screw thread and an oil-resistant pipette.

Preparation of drugs. Levodopa-PC nasal drops with the optimal composition shown in Table 3

A composite emulsion of L-DOPA lyophilizate (Sigma) as a part of polymer particles based on a biodegradable copolymer of lactic and glycolic acids (PLGA (Plurac Biochem), 50/50) and excipients (Levodopa substance). The particle size was 250 ± 50 nm, the inclusion of the active substance was 10 ± 2%. For administration to Wistar rats, the standard preparation was dissolved in a 1% ascorbic acid solution to a concentration of 3.2 mg / ml, initially adjusting the pH to 2 by adding a solution of 1M HCl. After dissolving L-DOPA, the pH was adjusted to 7 using 5M NaOH. Immediately prior to administration, the resulting Levodopa-PC formulation was adjusted to the desired concentration (by L-DOPA) to prevent the release of the active substance before it enters the body.

Research in the test of spontaneous activity of the forelimbs

This test was presented 4 weeks after the administration of 6-OHDA to trained animals (n = 112) with confirmed development of PD to search for a range of effective doses for their subsequent use with intranasal administration. The control was rats (n = 7), operated according to the indicated scheme, which were not injected with 6-OHDA, but received an equivalent volume of sterile saline.

Drug Administration and Dosage

All drugs were administered daily, nasally, using an automatic pipette. The volume of administration was 40 μl (20 μl in each nasal passage), the calculation of the presented doses was made in terms of the active substance. To control the physical condition of the animals, weekly weighing was performed. The effectiveness of a single injection of the test drug Levodopa in doses of 0.1-1 mg / kg (in terms of the active substance) and the comparison drug L-DOPA (prototype) in the same doses was evaluated when testing animals in the following mode: immediately before drug administration, through 30 min, after course administration after 3 days and after 7 days of daily nasal administrations of the compared drugs.

Example 1. Registration of spontaneous activity of the forelimbs

This test was presented 4 weeks after the administration of 6-OHDA to search for a range of effective doses with a view to their subsequent use with intranasal administration. The results obtained in the test of spontaneous activity of the forelimbs are summarized in Table 4.

The results of testing animals after daily administration of drugs for 3 days showed that in the entire range of doses tested, the rate of approximation to the norm after administration of Levodopa was always greater than in the control group without treatment for PD. The excess over the results of the prototype group was observed at doses not less than 0.2 mg / kg (D> 0.3 mg / kg). The test results of control animals with PD significantly differed from the results of the operated control group (p <0.05).

Dosages of 0.3-0.5 mg / kg can be considered optimal for the treatment of model PD with a single and course three-day administration, since an increase in doses to 1.0 mg / kg, i.e. 2-3 times, was not accompanied by a further increase in the percentage of correct decisions. Data obtained after daily administration of drugs for 7 days confirm this conclusion. Positive results in the aftereffect of administering the compared drugs were more pronounced and had a smaller scatter with the administration of the drug Levodopa-PK than with the introduction of the prototype L-DOPA, in the entire range of tested doses of 0.1-1 mg / kg

* P <0.05

Along with this, it should be noted that the therapeutic effect of the seven-day course of treatment was accompanied by an increase in the rate of correct reactions (touching the walls of the cylinder) to the “normal” level, identified in the “operated control” group, to the level of rats that were not administered 6-ONDA and, in in this sense, according to the safety of the dopaminergic system - to the level of intact animals. This result was observed in almost the entire range of tested doses of 0.1-1 mg / kg and persisted 24 hours after the last injection of Levodopa-PK, but not the prototype L-DOPA, which indicates a high specific activity and a clear therapeutic effect of the developed product . The range of tested doses of 0.1-1 mg / kg can be considered as therapeutic or as therapeutic breadth of the drug Levodopa-PK, as it was accompanied by a positive effect at low doses (0.1-0.3 mg / kg) and good tolerance at higher doses (0.35-1 mg / kg). However, according to a set of indicators: the necessary and sufficient effectiveness with a single (single) and multiple course administration, good tolerance, prolongation of the therapeutic effect after canceling the course of therapy, as well as technical difficulties arising from fractional administration of doses of large 0.4 mg / kg, for further experimental the study selected a dose of 0.35 mg / kg. It should be noted that this dose when transferring its value according to the rules of interspecific transfer from rat to human will correspond to the average value of a single therapeutic dose recommended for humans.

Example 2. Research in the test "Setting limbs"

In the course of the experiments, drug administration began from the 5th week after surgery, after confirming the development of PD in rats. The animals of the control groups were injected with an unloaded L-DOPA emulsion. The animals were divided into 4 groups (n = 10) based on the results of the “Limb Setting” test so that the average test results did not differ for all groups with PD.

All drugs were administered daily, nasally, using an automatic pipette. The volume of administration was 40 μl (20 μl in each nasal passage), the calculation of the presented doses was made in terms of the active substance. To control the physical condition of the animals, weekly weighing was performed.

The test "Setting the limbs" was carried out weekly during the entire period of drug administration. Animals were tested twice - immediately before drug administration and 30 minutes after their administration. Testing of animals in this mode with chronic administration made it possible to identify the effectiveness of the drug immediately after administration and to study the effect of the drugs one day after administration.

5 weeks after surgery, before the introduction of drugs, the results of testing groups of animals with induced PD did not exceed 75% of the results of the intact control group, the differences were significant (p <0.05).

The effectiveness of a single administration of the test drug Levodopa-PK at a dose of 0.35 mg / kg (in terms of the active substance) and the comparison drug L-DOPA (prototype) at a dose of 0.35 mg / kg was investigated when testing animals immediately before drug administration, through 30, 60 and 180 minutes after drug administration. The results are summarized in Table 5.

From the presented data it can be seen that 30 minutes after the administration of the drugs, the average number of successful attempts at setting the limb was 100 ± 4% for the intact control, 69 ± 14% for the control of PD, 88 ± 5% for the group receiving L-DOPA and 73 ± 21 % for the Levodopa-PK group One hour after administration, the results were as follows: 100 ± 5%, 66 ± 180%, 82 ± 90% and 75 ± 19%, respectively.

P <0.05

Significant differences were identified between the results of the BP control group and the group receiving L-DOPA (p <0.05), 30 and 60 minutes after drug administration. 180 minutes after the administration of the drugs, the results of all groups of animals with PD did not differ from each other, but in the Levodopa-PK group there was a trend towards an increase in the number of successful attempts to set the limb. Thus, the effect of a single administration of a standard preparation lasted no more than an hour, the effect of Levodopa-PK did not lead to significant changes in the coordination of rats. Differences in the effect of the studied drugs can be explained by the slow release of the drug from the polymer particles of Levodopa-PK, which does not allow to achieve an effective concentration with a single injection. At the same time, the effect of the comparison drug significantly increases the number of successful attempts when performing the test 30 minutes after the administration of L-DOPA with the complete disappearance of the effect by 180 minutes.

Testing of animals after daily administration of drugs for 7 days was carried out according to the same scheme as after a single administration — immediately before administration of the drugs, then 30, 60 and 180 minutes after their administration. On average, the number of successful attempts at setting a limb before drug administration was 100 ± 6% for intact control, 60 ± 17% for BP control, 65 ± 15% for the group receiving L-DOPA, and 80 ± 19% for the Levodopa-PK group. The test results of the control animals with PD significantly differed from the results of the intact control group (p <0.05). Significant differences were identified between the BP control group and the group receiving Levodopa-PK (p <0.05). The results of the study are recorded in Table 6.

It is important to note that in all studied time intervals after drug administration, significant differences in the test results were found only after administration of Levodopa-PK. In the group of animals treated with L-DOPA, there were no significant differences in test results from those in the BP control group. The leveling effect of the introduction of the comparison drug can be explained as follows: the daily dose of the standard drug L-DOPA in this experiment is 0.35 mg / kg, which is 60 times lower than the minimum therapeutic daily dose (in terms of person). It can be assumed that we observe a dose-depletion effect characteristic of levodopa therapy, which is one of the key problems of the use of the drug in the clinic and manifested itself so quickly due to the use of small doses of drugs.

P <0.05

It is important to note that the administration of Levodopa-PC for 7 days led to improved coordination of movements in animals with PD up to the next administration of the drug, that is, the effect persisted for 24 hours after the previous administration. Moreover, the effect was maximum within 180 minutes after administration (89% of the result of intact control), but somewhat decreased 24 hours after administration (83% of the result of intact control during testing before the next administration of the drug). The results are summarized in Table 7.

P <0.05

Testing of animals after a chronic course of daily administration of drugs for 120 days revealed a significant decrease in the ability to perform the test in the control group of animals with PD up to 51 ± 12% of the intact control. A similar dynamics was observed for the results of the group of animals treated with L-DOPA: by the 120th day of treatment, the test results 30 minutes after drug administration amounted to 62 ± 13% of the result of the intact control group. At the same time, the results of the group receiving Levodopa-PC were 94 ± 5% and were significantly higher compared with animals of both the BP control group (p <0.05) and the group receiving the prototype drug, which was observed throughout the entire period daily therapy.

As shown by the analysis of test data and observations obtained during the 180-day course of therapy, after a week of daily administration of Levodopa-PC, the advantages of the developed tool showed up in comparison with the standard preparation L-DOPA. Perhaps the differences in effect are associated with stabilization of the concentration of the drug encapsulated in PLGA at the therapeutic level. The coordination of movements in animals in the Levodopa-PK group was significantly better compared with animals in the BP control group (p <0.05) starting from the 7th day of therapy and throughout the rest of the drug administration. From the 14th day of therapy and on the following days, the results in the Levodopa-PK group also significantly differed from the results of the L-DOPA group (p <0.05). In this case, differences were observed both after 30 minutes and 24 hours after drug administration. After three weeks of daily use of Levodopa-PC, 30 minutes after drug administration, the ability of animals to perform the test did not significantly differ from the results of animals of the intact control group. The absence of differences persisted until the drug was discontinued, which indicates the complete restoration of the studied motor functions of animals after the administration of Levodopa-PC in the composition of polymer particles, but not of the prototype - L-DOPA. It is important to emphasize once again that the pronounced therapeutic effect of Levodopa-PK was observed not only immediately after administration of the drug, but also 24 hours after taking the drug. This fact indicates compensation for dopamine deficiency and stabilization of its level in the brain. In addition, the effect of the drug persisted for at least a week after its discontinuation and completely disappeared only a month after discontinuation of therapy. The experimental results are summarized in Table 8.

Example 3. The study of animals in the test "Open field"

The Open Field test was performed twice - immediately before the start of therapy and 6 weeks after the start of therapy. This test is used to assess the possible consequences of drug administration on the motor and research activity of experimental animals under conditions of free behavior in extreme conditions for them. The behavior of rats was studied in an "open field" in accordance with generally accepted standards. “Open field” - a square uniformly lit area 100 × 100 cm in size, fenced with opaque walls 40 cm high. The floor of the “open field” is laid out in squares.

The Open Field methodology is a classic testing system for studying animal behavior in an unfamiliar environment, based on a conflict of two dominant motivations in an unfamiliar space - an instinctive tendency to explore a new environment and a tendency to minimize the possible danger from it. The Open Field test is an informative technique that allows, in particular, to adequately assess the neurotropic, psychostimulating effects of pharmacological agents. This test assesses motor and orientational-research activity, as well as the emotional reaction of fear that occurs when studying an unfamiliar space.

For testing, rats were placed one at a time for 2 minutes in the center of a brightly lit, ragged, round platform. The following indicators were taken into account, horizontal activity (the number of sectors crossed), vertical activity (number of racks), acts of reaching the center of the field, the number of grooming, peeping, and the number of fecal boluses. The ratio of recorded indicators in the control and test groups was judged on the nature of the influence of substances on the psycho-emotional state (anxiety, fear) and on the level of tentatively research activity. An increase in anxiety is reflected in an increase in the number of fecal boluses and a decrease in research activity indicators. Before testing in the "open field" 60 minutes before testing, the animals were placed in a quiet, dimly lit room. During this period, any active manipulations were ruled out.

Before treatment, the experimental groups of animals did not differ in any of the recorded parameters. Testing 6 weeks after the start of therapy revealed a significant increase in the number of racks in the group of animals receiving Levodopa-PK compared with both the intact control and the control group with PD up to 162 ± 75% (p <0.05). A significant increase in the vertical activity of animals indicates an increase in research activity, a decrease in anxiety and an improvement in the general emotional state of animals. The introduction of the drug L-DOPA in the same dose did not lead to a significant change in the activity of animals. In addition, a significant decrease in the average number of grooming acts was revealed in the group of animals receiving Levodopa-PK compared with the group of intact control (p <0.05). The average value of the number of grooming acts for the Levodopa-PC group was 22 ± 23% of the result of intact control. The introduction of L-DOPA in the same dose did not lead to a significant change in the number of grooming acts in animals. Since there were no significant differences between the control groups during the experiment, the observed significant change in the number of grooming acts in the group receiving the drug Levodopa-PK should be attributed to the drug’s own action, which is not related to the development of BP, leading to a decrease in anxiety and faster decision-making in new for animal conditions.

Example 4. The study of animals in the test "Elevated cruciform maze"

The “Elevated Cruciform Maze” test and installation are designed to study the behavior of rodents under conditions of variable stress (with a free choice of comfortable conditions) and allows you to evaluate: the level of anxiety of the animal (according to the preference of darkness / light, fear of height, severity and dynamics of the “peeping” behavior); symptoms of neurological deficit; habituation.

The “Elevated Cruciform Maze” test was performed once, 11 weeks after the start of therapy. The study revealed significant differences only between the control groups of BP and the group receiving Levodopa-PC in reducing the number of auto-grooming acts to 50% of the value of the intact control group (p <0.05), which reproduces the results obtained in the Open Field test. A sharp decrease in the level of auto-grooming in the group receiving Levodopa-PC along with an increase in the level of research activity in the open field and with a decrease in the level of bowel movements is considered as a suppression of the level of anxiety and fear.

The test results of Levodopa-PC in the tests “Open Field” and “Elevated Cruciform Labyrinth” allow us to conclude that a comprehensive increase in the research components of behavior and a decrease in psychoemotional stress and fear in a stressful environment indicate the restoration of dopaminergic functions under the influence of Levodopa-PC, which talks about the pharmacological effect of the tested dose. A comparative assessment of the effect of the drugs showed that the detected shift in the direction of increasing psycho-emotional stability and enhancing research behavior is more pronounced with the nasal administration of Levodopa-PK than with the administration of the L-DOPA comparison drug.

There were no other significant differences in the behavior of animals by the parameters recorded in the test.

Example 5. Test "Movement on a horizontal grid"

Studies in the test "Movement on a horizontal grid" allow us to give a comparative assessment of the degree of preservation of locomotor and highly coordinated motor reactions in animals with damaged dopaminergic nigrostriatal system relative to the behavior of intact animals. The animals were placed one at a time in the center of the horizontal square grid-screen (1 × 1 m) for 4 minutes, the number of steps of each limb along the horizontal grid and the percentage of errors during the setting of each limb were determined.

A single study of animals was carried out after 30 and 60 days of therapy with an analysis of the number of steps of each limb on a horizontal grid, the percentage of errors in the formulation of each limb, and the asymmetry coefficient 30 minutes after drug administration.

The number of steps by both the left and right foot did not significantly differ in animals of all groups. The percentage of errors in the setting of the left limb was 7.5 ± 4.8% for intact control, 14.2 ± 6.7% for control of PD, 12 ± 5.9% for the group receiving levodopa, and 9.9 ± 3, 4% for the Levodopa-PK group. The percentage of errors in the BP control group significantly differed from that in the intact control by half (p <0.05), however, the results of the group receiving Levodopa-PC, as well as the group receiving the comparison drug, did not significantly differ from the intact control. When examining the percentage of errors in setting the right limb, no significant differences between the groups were found.

The asymmetry coefficient 30 minutes after drug administration was 0.9 ± 4.6% for intact control, 5.8 ± 4.9% for control of PD, 6.7 ± 3.3% for the group receiving levodopa, and 1, 3 ± 4.3% for the Levodopa-PC group (absolute values of the parameter are presented). The asymmetry coefficients of the BP and L-DOPA control groups significantly differed from that of the intact control group (p <0.05) and the Levodopa-PC group. The asymmetry coefficient of the group of animals treated with Levodopa-PK was closest to the value of the intact control group and significantly differed from the control of BP and the L-DOPA group.

The obtained results of the test "Movement on a horizontal grid" are completely consistent with the results obtained in the test "Setting limbs" and confirm both the presence of motor dysfunctions in animals with induced PD and significantly higher efficacy of nasal therapy of Levodopa-PC compared to the standard drug in the same dose.

The results of the experiment are summarized in Table 9.

In all tests performed, the absence of significant differences during manipulation of the right limb is explained by the unilateral nature of the damage s. nigra in the used model of PD, which mainly affects the capabilities of the opposite damaged part of the brain of the side of the body.

The technical result established the prolonged action and bioavailability, namely, after obtaining and experimental substantiation of the BP model, caused by the intracerebral administration of 6-hydroxydopamine to rats. The new pharmaceutical composition showed the following.

- Intranasal administration of Levodopa-PC in doses of 0.1-1.0 mg / kg according to L-DOPA led to a significant improvement in the coordination of movements of animals with PD in the test “Cylinder-contact of the forelimbs” through three- and seven-day courses of daily nasal administration. The optimal therapeutic dose of treatment for this test has been established.

- The course intranasal administration of Levodopa-PC at a dose of 0.35 mg / kg according to L-DOPA led to a significant improvement in the coordination of movements of animals with PD after a week of daily nasal administration. The therapeutic effect was maintained throughout the entire period of administration, starting from day 7, and a week after discontinuation of the drug.

- The prolonged action of Levodopa-PK was found, which was expressed in maintaining the effectiveness of the drug 24 hours after a single injection and throughout the treatment period, in contrast to the standard drug, the effectiveness of which decreased with the course of the course.

- The detected effect of Levodopa-PK on the psycho-emotional state of animals, leading to a decrease in anxiety and an increase in research interest, is the drug's own effect and can be useful for improving the well-being of patients in the treatment of PD.

Claims (9)

1. A pharmaceutical composition for the treatment of Parkinson's disease in the form of nasal drops containing 3.4 dihydroxy-L-phenylalanine (L-DOPA), a biodegradable copolymer of lactic / glycolic acid polylactide glycolide 50/50 (PLGA 50/50), cryoprotectant D-mannitol, polyvinyl alcohol (PVA), olive oil, Tween 80 in the following amount, g: 3.4 dihydroxy-L-phenylalanine - 0.5 polylactide glycolide 50/50 - 4 D-mannitol - 0.4 polyvinyl alcohol - 0.1 olive oil - 4.9 Twin 80 - 0.1 2. A method of obtaining a pharmaceutical composition according to claim 1, comprising obtaining a PVA solution of saturated L-DOPA at a temperature of 37-42 ° C with constant stirring until the solid phase is completely dissolved; obtaining a suspension of L-DOPA in a solution of a copolymer PLGA 50/50 in dichloromethane; obtaining an emulsion by adding to a solution of PVA saturated with L-DOPA a suspension of L-DOPA in a solution of a copolymer of PLGA 50/50 in dichloromethane in a water bath at a temperature of 21-27 ° C and stirring, stop heating, drain the water and fill the tank with ice ; carry out the second emulsification - homogenization to a foamy mass, conduct ultrasonic dispersion, evaporate dichloromethane until a vacuum of 0.93-0.98 units is achieved .; the resulting suspension composition of L-DOPA with a PLGA 50/50 copolymer is mixed with a cryoprotectant D-mannitol dissolved in water, filtered, frozen, lyophilized; olive oil is added to Tween 80 and mixed, then lyophilized polymer particles of L-DOPA in a PLGA 50/50 copolymer are added to the resulting mixture and mixed at a speed of 50-60 rpm for 5-10 minutes with a uniform distribution of the oil mixture in the entire volume lyophilized particles to form a homogeneous mass. 3. The use of the pharmaceutical composition obtained by the method according to claim 2, for the treatment of Parkinson's disease.