RU2697254C1 - Pharmaceutical composition based on bis-(n-monosuccinyl-l-seryl-l-lysine) hexamethylenediamide (gsb-106) - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention refers to medicine, particularly to pharmacy, and concerns a pharmaceutical composition containing a dimeric dipeptide mimetic BDNF - hexamethylene diamide bis-(N-monosuccinyl-L-seryl-L-lysine) (GSS-106) and as additives - filler, binder and glidant, in amounts specified in claims. Preferably, the filler used is a mixture of lactose and microcrystalline cellulose with particle size of 101 in ratio of 2:1, respectively, as a binder, a copolymer of polyvinyl alcohol and polyethylene glycol, as a sliding substance - magnesium stearate. Pharmaceutical composition is intended for forming tablets and filling capsules.EFFECT: pharmaceutical composition under the invention extends the range of products with antidepressant activity in a solid dosage form and meets all requirements of GF XIII.3 cl, 4 tbl, 4 ex

Description

The invention relates to medicine, in particular to pharmacy, and relates to a pharmaceutical composition containing bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylenediamide (GSB-106) in the form of a solid dosage form having antidepressant activity.

Depression is the most common mental illness. According to WHO, depression affects 4-5% of the world's population. According to various sources, from 5 to 15-40% of children and adolescents aged 10 to 16 years are depressed. By 2030, depression may come first in terms of prevalence among other diseases.

The antidepressants on the pharmaceutical market are inhibitors of monoamine oxidase or reuptake of monoamines - serotonin, dopamine, adrenaline; their action is aimed at restoring the normal level of monoamines. A positive effect is achieved only with prolonged use of these drugs, while from 30 to 50% of patients with depression do not respond to antidepressants. In addition, prolonged use of antidepressants is accompanied by a risk of side effects from the central and autonomic nervous system [Vereitinova VP, Tarasenko O.A. Side effect of antidepressants // Pharmacist. 2003. No. 14; Martsenkovsky I.A. Clinical pharmacology of antidepressants: mechanisms of effectiveness and side effects // Neuro News: neuropsychiatrics and neuropsychiatry. 2006. No. 1].

Experimental and clinical data indicate that the development of symptoms of a depressed state is associated with impaired neurogenesis and neuroplasticity in the hippocampus and is associated with severe deficiency of the brain-derived neurotrophic factor BDNF [Dwivedi Y. Involvement of Brain-Derived Neurotrophic Factor -Life Depression // Am. J. Geriatr. Psychiatry. 2013. V. 21, No. 5. P. 433-449] - a protein that is an endogenous regulator of neurogenesis and synaptogenesis [Jiang C, Salton R. The Role of Neurotrophins in Major Depressive Disorder // Transl Neurosci. 2013. V. 4, No. 1. P. 46-58].

To date, a large amount of data has been accumulated indicating the central role of BDNF deficiency in the pathogenesis of depression. Clinical studies have shown that plasma BDNF levels decrease in people with depression and return to normal after treatment with antidepressants [Jiang C, Salton R. The Role of Neurotrophins in Major Depressive Disorder // Transl Neurosci. 2013. V.4, No. 1. P. 46-58]. Post-mortem analysis of suicide victims reveals a sharply reduced BDNF content in the prefrontal cortex and hippocampus [Karege F., Vaudan G., et al. Neurotrophin levels in postmortem brains of suicide victims and the effects of antemortem diagnosis and psychotropic drugs // Molecular Brain Research. 2005. V. 136, No. 1-2. P. 29-37]. In experimental models of depression, it was found that BDNF with intracerebral administration has a pronounced antidepressant effect [Shirayama Y, Chen AC, Nakagawa S, et al. Brain-derived neurotrophic factor produces antidepressant effects in behavioral models of depression // J. Neurosci. 2002. V. 22. P. 3251-3261]. However, the therapeutic use of native BDNF is limited by its instability in biological fluids and the low ability to systemically penetrate the gastrointestinal and blood-brain barrier [Monteggia LM. Toward neurotrophin-based therapeutics // Am J Psychiatry. 2011, 168: 114-6].

One solution to the above problems is the use of short, mainly di - and tetrapeptides. Short peptides are able to penetrate biological barriers due to both passive and active transport [Bradbury M. The concept of the blood-brain barrier. M. 1983; 480 s .; Ryabukhin I.A., Dmitrieva T.B., Chekhonin V.P. The blood-brain barrier. Embryomorphogenesis, cell and subcellular biology of close contacts of endotheliocytes. Neurochemistry. 2003. No.20. S. 12-23.]. In addition, they are characterized by increased enzymatic stability, which allows them to create medications for intragastric administration [Ostrovskaya R.U., Gudasheva T.A., Voronina T.A. et al. The original nootropic and neuroprotective drug Noopept // Experiment, and wedge, pharmac. 2002, T. 65. S. 66-72]. However, low molecular weight peptides exhibit high activity, which is manifested by such serious undesirable side reactions as water retention in the body and increased blood pressure [EB-M Daliri, DH Oh, VN Lee. Bioactive peptides. Review / MDPI Foods, 2017. No. 6 (5), P.32; Petrov B.H., Rybakov C.C., Petrova O.H. et al. Synthesis and immunogenic properties of peptides - fragments of immunodominant regions of the VP1 protein of foot and mouth disease virus of type Asia-1 // Bioorgan, chemistry. 1998. V. 24. No. 12. C 899-905; Havinson V.Kh. Peptide regulation of aging / St. Petersburg. "The science". 2009.56 p.].

To solve the problem of effective and safe treatment of depression, an innovative antidepressant is proposed, the first representative of a new class of drugs - low molecular weight BDNF mimetics. The new molecule underlying the drug is the dimer dipeptide mimetic BDNF, hexamethylene diamide bis (N-monosuccinyl-L-seryl-L-lysine (GSB-106). GSB-106 was created based on the beta bend of the 4th loop of BDNF in the Department of Chemistry of Medicines of the VV Zakusov Research Institute of Pharmacology under the supervision of T. A. Gudasheva, Corresponding Member of the Russian Academy of Sciences [T. Gudasheva, A. V. Tarasyuk, S. V. Pomogaybo, I.O. Logvinov, Povarnina P.Yu., Antipova T.A., Seredenin S.B.Design and synthesis of dipeptide mimetics of brain neurotrophic factor // Bioorganic chemistry. 2012.V. 38 , No. 3. P. 280-290; RF Patent No. 2410392, 2010; US Patent US 9683014 B2, 2017; China Patent CN 102365294 B, 2016]. Neuropsychotropic properties and the mechanism of action of GSB-106 were investigated under the supervision of Academician RAS SB. Seredenin.

GSB-106 was shown to activate BDNF-specific tyrosine kinase TrkB receptors, their main signaling pathways - PI3K / AKT and MAPK / ERK [Gudasheva TA, Logvinov IO, Antipova TA, Seredenin SB. The dipeptide mimetic of the 4th loop of brain neurotrophic factor GSB-106 activates TrkB, Erk, and Akt and promotes neuronal survival in vitro // DAN. 2013.Vol. 451, No. 5. S. 577-580]. In vitro studies of GSB-106 on a culture of immortalized mouse hippocampal HT22 line cells showed that at concentrations of 10 ^-5 -10 ^-8 M the compound exhibits neuroprotective activity under conditions of oxidative stress and glutamate toxicity. The neuroprotective effect of GSB-106 was also detected on cells of the SH-SY5Y line of human neuroblastoma under the action of neurotoxin 6-oxidophamine [Logvinov I.O. et al. Neuroprotective properties of the dipeptide mimetic of brain neurotrophic factor GSB-106 in in vitro experiments // Bulletin of Experimental Biology and Medicine. 2013.V. 155, No. 3. S. 319-322].

GSB-106 was found to exhibit antidepressant activity in doses of 0.1-1.0 mg / kg (intraperitoneal) with a single and subchronic administration in a number of rodent tests: unavoidable swimming in Porsolt, forced swimming in Nomur and suspension by tail in Ster [Gudasheva T.A. et al. Design and synthesis of dipeptide mimetics of brain neurotrophic factor // Bioorganic chemistry. 2012.V. 38, No. 3. S. 280-290; Seredenin S.B. et al. Antidepressant effect of the original low molecular weight BDNF mimetic, GSB-106 dimeric dipeptide // Acta Naturae. 2013.Vol. 5, No. 4. S. 116-120]. Moreover, GSB-106 was not inferior to the classical antidepressant amitriptyline in terms of severity of effects (10 mg / kg, intraperitoneally). GSB-106 was shown to have a stimulating effect on neurogenesis under conditions of subchronic stress in mice caused by contact with a predator [Gudasheva T.A. et al. The dipeptide mimetic of cerebral neurotrophic factor prevents neurogenesis impairment in stressed mice // Bull. expert. biol. and honey. 2016.V. 162. No. 10. S. 448-451]. Preliminary toxicological and pharmacokinetic studies have shown that GSB-106 is non-toxic (LD50 for outbred male mice> 4.5 g / kg) and crosses the blood-brain barrier.

Peptide preparations available on the pharmaceutical market mainly imply a parenteral or local route of administration (intramuscular, intranasal, etc.), which is particularly painful [Yin N., Brimble MA, Harris P.W.R. et al. Enhancing the Oral Bioavailability of Peptide Drugs by using Chemical Modification and Other Approaches / Medicinal Chemistry, 2014. V.4 (12). P. 763-769]. It is known that the frequency of refusals from treatment among mental patients is higher than in surgery and cardiology, where approximately every seventh patient refuses the full amount of therapy offered to him. Most of all, this problem concerns patients who are forced to take drugs for a long time or for life [Krizhanovsky A.S., Ivanov M.V., Maso G.E. New possibilities for solving the problem of compliance // Review of psychiatry and medical psychology named after B.M. Ankylosing spondylitis, 2010. No. 1, pp. 48-51]. Therefore, it is preferable to obtain a pharmaceutical composition in solid dosage form, in particular a tablet, which has great advantages over other dosage forms, among which the most important are painless administration, ease of use, which ensures patient compliance, as well as greater stability and long shelf life the drug.

Thus, the development of low molecular weight peptide highly effective drugs for the treatment of mental illness, the administration of which is convenient to use, painless and does not cause side effects is relevant.

The objective of the present invention is to develop a new highly effective pharmaceutical composition based on bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene diamide (GSB-106), made in the form of a solid dosage form, mainly tablet, characterized by pronounced antidepressant activity.

The technical result of the invention is the expansion of the range of convenient in use and storage of drugs with antidepressant activity in tablet dosage form, with high bioavailability and improved therapeutic efficacy and the absence of side and unwanted effects.

The solution to this problem was achieved by creating a pharmaceutical composition that meets all the requirements of the State Pharmacopoeia XIII edition of a pharmaceutical composition containing a therapeutically effective amount of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene in combination with the following excipients in amounts that are mainly in the following ranges :

Cellulose derivatives, sugars can be used as a filler of said pharmaceutical composition providing the necessary weight of a tablet or capsule; preferably microcrystalline cellulose and lactose; most preferably a mixture of lactose and microcrystalline cellulose with a particle size of 101 in a ratio of 2: 1, respectively.

Derivatives of polyvinyl alcohol, preferably a copolymer of polyvinyl alcohol and polyethylene glycol (Kollicoat IR), can be used as a binder in this pharmaceutical composition.

As a solvent for preparing the binder solution, it is preferable to use purified water.

As a sliding substance that can eliminate the problems associated with the formation of tablets, for example, the sticking of the tablet mass on the punches during tabletting, mainly the use of derivatives of higher fatty acids and their salts, such as, for example, stearic acid and its salts, preferably magnesium stearate.

The pharmaceutical composition is in the form of a solid dosage form by any suitable method, mainly in the form of a tablet containing 0.12-12.5% of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene diamide.

For the manufacture of tablets, the use of wet granulation technology is predominant.

The technological process includes the following stages: weighing, sieving, mixing, preparation of a binder solution, moistening, wet granulation, drying, dry granulation and dusting.

During the development of the pharmaceutical composition, experimental compositions were obtained that qualitatively differ in the type of binder selected (Table 1). So, as a binder when creating a new pharmaceutical composition based on GSB-106, we used:

1) pregelatinized starch (Starch 1500),

2) polyvinylpyrrolidone (Kollidon 25) with a molecular weight of 28000-34000,

3) a grafted copolymer of polyvinyl alcohol and polyethylene glycol (PVA-PEG) (Kollicoat IR).

The following characteristics of the tablet mixture and tablets were studied: flowability, bulk density, crushing strength, disintegration. The measurement of technological characteristics was carried out in accordance with the requirements of the Global Fund of the 13th edition of Volume 2. Based on the results obtained using the generalized Harrington function, unexpectedly, the best value of the indicator was obtained for composition No. 3, containing Kollicoat IR as a binder. The results are presented in table 2:

и индекса

, которые при использовании технологии влажного гранулирования имеют определяющее значение, поскольку влияют на временные сроки технологического процесса при масштабировании.From the above data it is seen that the best composition is composition No. 3, which has a significant advantage in terms of bulk density, index

and index

which, when using wet granulation technology, are of decisive importance because they affect the timelines of the process during scaling.

The data obtained indicate a positive effect of Kollicoat IR on the technological properties of both the powder mixture and tablets.

и

.When comparing the technological parameters of the substance GSB-106 and the tablet mixture No. 3 on the basis of GSB-106 using Kollicoat IR, an improvement in the technological characteristics of the tablet mixture was found for all recorded indicators: flowability, bulk density, indices

and

.

* ¹ - characterizes the degree of flowability of powders; considered satisfactory at a value of less than 20%;

* ² - characterizes the degree of flowability of powders; an index value of less than 1.25 indicates satisfactory flowability and compressibility of the tablet mixture.

The implementation of the invention is demonstrated by the following examples, not limiting it:

Example 1

1. Weighing and sieving of raw materials. Weigh out 330.00 g of microcrystalline cellulose 101, 600.0 g of lactose, 50.0 g of Kollicoat IR, 10.0 g of magnesium stearate and 10.0 g of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene.

2. Preparation of a 10% solution of Kollicoat IR: measure 440.0 g of purified water, add 50.0 g of Kollicoat IR, mix until the Kollicoat IR is completely dissolved. To the resulting solution, with constant stirring, 10.00 g of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene diamide are added until the substance is completely dissolved.

3. Moisturizing. The resulting solution is moistened with a pre-prepared mixture consisting of 330.00 g of microcrystalline cellulose 101 and 600.00 g of lactose.

4. Wet granulation. The moistened mixture is punched through a sieve with a hole diameter of 2 mm.

5. Drying. The obtained granulate is dried in an oven at t = 40-45 ° C until complete drying.

6. Dry granulation and dusting. The dried mixture is punched again through a sieve and dusted with 10.00 g of magnesium stearate for 1-3 minutes.

7. Tableting. From the obtained granulate, tablets are obtained weighing 0.100 g in an amount of 10,000 tablets.

Example 2

1. Weighing and sieving of raw materials. 731.0 g of lactose, 60.0 g of Kollicoat IR, 8.0 g of magnesium stearate and 1.0 g of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene diamide are weighed out.

2. Preparation of a 12% solution of Kollicoat IR: measure 439.0 g of purified water, add 60.0 g of Kollicoat IR, mix until Kollicoat IR is completely dissolved. 1.00 g of bis- (N-monosuccinyl-L-serial-L-lysine) hexamethylene diamide are added to the resulting solution with constant stirring until the substance is completely dissolved.

3. Moisturizing. The resulting solution 731.0 g of lactose.

4. Wet granulation. The moistened mixture is punched through a sieve with a hole diameter of 2 mm.

5. Drying. The obtained granulate is dried in an oven at t = 40-45 ° C until complete drying.

6. Dry granulation and dusting. The dried mixture was punched again through a sieve and dusted with 8.00 g of magnesium stearate for 1-3 minutes.

7. Encapsulation. A mixture is obtained from the obtained granulate, which is filled with 0.080 g hard gelatin capsules in an amount of 10,000 capsules.

Example 3

1. Weighing and sieving of raw materials. 450.0 g of lactose, 895.0 microcrystalline cellulose 101, 40.0 g of Kollicoat IR, 15.0 g of magnesium stearate and 100.0 g of bis- (N-monosuccinyl-L-seryl-L-lysine) hexane are weighed out.

2. Preparation of an 8% solution of Kollicoat IR: measure 360.0 g of purified water, add 40.0 g of Kollicoat IR, mix until complete dissolution of Kollicoat IR. To the resulting solution, 100.00 g of bis- (N-monosuccinyl-L-serial-L-lysine) hexamethylene diamide are added with constant stirring until the substance is completely dissolved.

3. Moisturizing. The resulting solution is moistened with a pre-prepared mixture consisting of 895.00 g of microcrystalline cellulose 101 and 450.00 g of lactose.

4. Wet granulation. The moistened mixture is punched through a sieve with a hole diameter of 2 mm.

5. Drying. The obtained granulate is dried in an oven at t = 40-45 ° C until complete drying.

6. Dry granulation and dusting. The dried mixture is punched again through a sieve and dusted with 15.00 g of magnesium stearate for 1-3 minutes.

7. Tableting. From the obtained granulate tablets of 0.150 g in the amount of 10,000 tablets are obtained.

Tablets or capsules obtained from various compositions of the invention contain from about 0.0001-0.010 g of bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylene diamide per tablet. The weight of the obtained tablet is preferably from 0.080 to 0.150 g.

Example 4

The study of the antidepressant activity of the finished dosage form (LF) based on bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylenediamide obtained by the method described in example 1.

The study of the antidepressant activity of the pharmaceutical substance GSB-106 was carried out on outbred male rats weighing 270-290 g, obtained from the Stolbovaya Central Laboratory Animal Laboratory Kennel (Moscow Region). The organization and conduct of work was carried out in accordance with the order of the Ministry of Health of Russia No. 199 of April 1, 2016 “On approval of the rules of good laboratory practice”. The animals were kept in accordance with SP 2.2.1.3218-14 "Sanitary and epidemiological requirements for the design, equipment and maintenance of experimental biological clinics (vivariums)" dated August 29, 2014 No. 51.

GSB-106 (substance) was diluted in distilled water and was administered to rats daily for 14 days intragastrically using a special probe in doses of 0.1; 0.5; one; 5 and 10 mg / kg. Control animals received distilled water.

The dosage form GSB-106 was suspended in a 1% starch solution and was administered to rats daily for 14 days intragastrically using a special probe in doses of 0.001; 0.01; 0.05; 0.1 and 5 mg / kg (calculation based on the content of active substance).

Control animals (placebo group) received a filler dosage form (without the active substance - GSB-106), suspended in a 1% starch solution. The classical antidepressant amitriptyline at a dose of 5 mg / kg (intragastric, 14 days) was used as a comparison drug.

The antidepressant activity of GSB-106 was evaluated in the Porsolt test of unavoidable swimming [Porsolt BD, Bertin A, Jalfre M. // Eur. J. Pharmacol. 1978. V. 51. P. 291-294. Buccafusco JJ. Methods of Behavior Analysis in Neuroscience. 2nd edition, CRC Press, Florida. 2009]. To this end, 24 hours after the last injection of substances, rats were placed in cylinders with water (22 ° С) for 15 minutes. After 24 hours, the rats were placed under the same conditions for 5 minutes and the immobility time was recorded. A decrease in the duration of immobility was regarded as evidence of the antidepressant activity of the compounds.

Statistical analysis was performed using the Biostat program using the Mann-Whitney U-test. Differences were considered statistically significant at p≤0.05.

It is shown that GSB-106 in the substance in doses of 0.5; 1 and 5 mg / kg statistically significantly reduced the immobility time in rats in the Porsolt test of inevitable swimming, which indicates the manifestation of antidepressant activity. By its severity, the effect of GSB-106 in the substance was comparable with the effect of the classical antidepressant amitriptyline (Table 4). At doses of 0.1 and 10 mg / kg GSB-106 (substance) did not affect the immobility time. Thus, the range of active doses of GSB-106 (substance) was established in this test: from 0.5 to 5 mg / kg with intragastric administration.

* - p <0.05 - statistical significance of differences compared with the control group for the U Mann-Whitney test with Bonferroni correction. Data are presented as mean and standard error of the mean.

GSB-106 (dosage form) in doses of 0.001; 0.01; 0.05; 0.1 and 5 mg / kg (intragastric) statistically significantly reduced the immobility time in rats in the Porsolt test of unavoidable swimming (Table 4). The effectiveness of the dosage form at a dose of 0.001 mg / kg was comparable with the effectiveness of amitriptyline at a dose of 5 mg / kg (Table 4). The effectiveness of the GSB-106 dosage form at doses of 0.01, 0.005 and 0.1 mg / kg was more pronounced than that of amitriptyline, which was manifested in a decrease in the duration of rat immobility by 37%, 45% and 33%, respectively, compared with control, whereas against the background of amitriptyline, this indicator was less by 24.4% (table. 4).

Thus, the antidepressant activity of GSB-106 in tablet form is manifested in significantly lower doses (0.001-0.1 mg / kg, intragastrically) compared with GSB-106 in substance (0.5-5.0 mg / kg, intragastrically). Moreover, the effect of GSB-106 in a tablet dosage form at doses of 0.001-0.1 mg / kg was significantly more pronounced than that of the classical antidepressant amitriptyline (5 mg / kg).

The results obtained indicate an increase in the activity of the proposed pharmaceutical composition compared with the pharmaceutical substance hexamethylene diamide bis- (N-monosuccinyl-L-seryl-L-lysine), which is expressed in a decrease in the dose required for the manifestation of antidepressant action from 0.5-5 mg / kg to 0.01-0.1 mg / kg, as well as an increase in the severity of effects: the time of immobilization with the introduction of the dosage form GSB-106 is reduced by 33-45%, while with the introduction of a pharmaceutical substance - by 20-35%.

Claims (4)

1. A pharmaceutical composition for preparing a solid dosage form of bis- (N-monosuccinyl-L-serial-L-lysine) hexamethylenediamide, containing a therapeutically effective amount of bis- (N-monosuccinyl-L-serial-L-lysine) hexamethylene diamide) and the following groups of excipients in the indicated amounts (wt.%): Bis- (N-monosuccinyl-L-seryl-L-lysine) hexamethylenediamide 1,0 Filler 93.0 Binder 5,0 Sliding 1,0 2. The pharmaceutical composition according to claim 1, containing microcrystalline cellulose and lactose as a filler; most preferably a mixture of lactose and microcrystalline cellulose with a particle size of 101 in a ratio of 2: 1, respectively; as a binder, a copolymer of polyvinyl alcohol and polyethylene glycol (Kollicoat IR); as a glidant, magnesium stearate; purified water is used as a solvent for the preparation of a binder solution. 3. The pharmaceutical composition according to claim 1 is intended for forming tablets and filling capsules.