RU2326870C2 - Application of cyclic bioisoster derivaties of purine system for treatment of disorders caused by abnormalities of nitrergic and dopaminergic systems - Google Patents

Abstract

FIELD: medicine; veterinary science.

SUBSTANCE: invention refers to application of compounds with common structural formula

R¹=-H, -NH₂, -Br, -Cl, -OH, -COOH,

B=-N=, -CH=, Z=-CH=, -N=,

A=-CH- at B=-N=, Z=-CH-,

A=-CH- at В=-CH=, Z=-CH=,

A=-N= at B=-N=, Z=-CH-,

A=-CH- at B=-N=, Z=-N=,

A=-CH= at В=-CH=, Z=-N=.

Structures of specified formula are active for nitrergic and dopaminergic systems of mammal body including human body. These compounds can be applied as neuroprotectors, to improve cognitive function and to normalise psychophysiologic state, to treat consequences of substance abuse, as well as to treat wide range of diseases including neuropsychic, cardiovascular, immune, inflammatory and gastro-intestinal disorders.

EFFECT: application of new and well-known compound to effect nitrergic and dopaminergic systems for treatment purposes.

4 ex, 3 tbl, 8 dwg

Description

Technical field

The invention relates to medicine, in particular to medicines with a directed effect on the most important mediator systems of the body, in particular for the treatment of various diseases associated with disorders of the nitrergic and dopaminergic systems of the body. Such disorders include neurological, neuropsychic and cardiovascular diseases and disorders caused by substance abuse, in particular drugs, impaired cognitive function and psychophysiological state.

State of the art

It is known that a number of diseases of the nervous system and cardiovascular diseases, in particular the abuse of substances - drugs, alcohol, nicotine and other substances, as well as various mental disorders are associated with impaired functioning of the neurotransmitter systems. The state of these important systems substantially determines the psychophysiological state of a person and almost all the functions of the central and peripheral nervous system in normal and pathological conditions. These systems, in particular, include nitrergic and dopaminergic systems.

Nitric oxide NO - a low molecular weight gas of a free radical nature - easily penetrates cell membranes and components of the intercellular substance, playing an important role in a wide variety of physiological processes. The effect of NO on the state of the cell is largely dependent on the amount of gas. In small quantities, produced mainly by the neuronal and endothelial isoforms of NO synthase, the effect of NO is mainly associated with the effect on the heme group of the soluble form of the guanylate cyclase enzyme. The activated enzyme synthesizes cyclic guanosine monophosphate (hereinafter cGMP), which regulates the operation of membrane ion channels, protein phosphorylation, phosphodiesterase activity and other reactions. At high concentrations produced by the macrophage isoform of NO synthase, NO can have a toxic effect on cells, associated both with a direct effect on Fe-containing enzymes and with the formation of a strong oxidizing agent - the free radical peroxynitrite compound (ONOO ^- ), which is formed by the interaction of NO with superoxide radical (O ₂ ^- ). The toxic effect of nitric oxide is manifested primarily in the inhibition of mitochondrial enzymes, which leads to a decrease in the production of adenosine triphosphate (hereinafter ATP), as well as enzymes involved in DNA replication. The ability of peroxynitrite and directly NO to damage DNA leads to the activation of protective mechanisms, in particular, the activation of the poly (ADP-ribose) synthetase enzyme, which, in turn, leads to a decrease in ATP level and can lead to cell death.

NO is synthesized in a cell from L-arginine by the enzyme NO synthase (hereinafter NOC), which converts L-arginine to NO and citrulline. This synthetic pathway is implemented, in particular, in the cardiovascular and central nervous system (hereinafter CNS), where NO performs the function of a signaling molecule, including a neurotransmitter function. The neurotransmitter function of NO is that it is synthesized upon excitation of a neuron and, diffusing into neighboring cells, activates the formation of cGMP in them, which can affect the conductivity of ion channels, and thus change the electrogenesis of neurons. In addition, NO in the central nervous system provides a new, non-synaptic and not mediated by receptors, pathway of interneuronal communication (Kiss JP, Vizi ES, Nitric oxide: a novel link between synaptic and nonsynaptic transmission. Trends Neurosci., 2001, Apr., 24 (4): 211-5).

The influence of the nitrergic system on the functions of the central nervous system is carried out both directly and indirectly through other neurotransmitter systems. For example, glutamate and its receptors mediate the most important functions of the central nervous system, including memory, and also affect the development of depression and antidepressant activity. NO and nitric oxide synthase are important components of the signal transmission system in the glutamatergic synapse (Paul I.A., Skolnick P. Glutamate and depression: clinical and preclinical studies. Ann. N Y Acad. Sci., 2003, Nov; 1003: 250-72). The activity of monoamine oxidase, a key enzyme of monoamine metabolism in the brain, has been shown to be related to the amount of NO in the cell (Girgin Sagin F., Sozmen EY, Ersoz B., Mentes G. Link between monoamine oxidase and nitric oxide. Neurotoxicology, 2004, Jan., 25 (1-2): 91-9).

NO affects the function of monoaminergic transporters (Vizi E. S. Role of high-affinity receptors and membrane transporters in nonsynaptic communication and drug action in the central nervous system. Pharmacol. Rev. 2000, Mar. 52 (1): 63-89). NO facilitates the release of many monoamines, especially dopamine, and also blocks the presynaptic reuptake of dopamine if necessary. Thus, NO increases dopamine lifespan at the synapse. In connection with the participation of dopamine in motor and psychological processes, nitrergic influences on these processes are attracting increasing attention (Liu Y. Nitric oxide influences dopaminergic processes. Adv. Neuroimmunol., 1996, 6 (3): 259-64).

NO mediates many behavioral and neuroendocrine responses of the body, in particular aggressive and impulsive behavior. NO plays an important role in the functioning of serotonin receptors in the brain (Chiavegatto S., Nelson R. J. Interaction of nitric oxide and serotonin in aggressive behavior. Horm. Behav. 2003, Sep. 44 (3): 233-41). Monoaminergic systems and the NO system of the hypothalamus, limbic and stem structures participate in the regulation of sexual behavior, control partner preference, sexual desire, erection, copulation, ejaculation, body and sexual satisfaction) (Pfaus JG Neurobiology of sexual behavior. Curr. Opin. Neurobiol. 1999, Dec. 9 (6): 751-8).

It is known that NO plays an important role in the formation of dependence on various drugs, including opioids, ethanol, psychostimulants and nicotine. In particular, NO is involved in the manifestation and development of symptoms of drug withdrawal. For example, activation of the opiate receptor μ3 is accompanied by the release of NO in endothelial cells, granulocytes, monocytes and microglia (Stefano GB Autoimmunovascular regulation: morphine and anandamide and ancondamide stimulated nitric oxide release. J. Neuroimmunol., 1998, Mar., 15, 83 (1- 2): 70-6), NO plays an important role in the development of dependence on compounds of various classes causing dependence. Thus, modulation of the NO system can be a potential therapeutic target for the treatment of various kinds of addictions (Tayfun Uzbay I., Oglesby MW Nitric oxide and substance dependence. Neurosci. Biobehav. Rev., 2001, Jan., 25 (1): 43-52 )

NO is involved in the regulation of neurotransmission in the central nervous system, in particular, mediating non-synaptic interactions, regulating monoaminergic systems - dopaminergic, noradrenergic. Thus, dysfunction of the NO system is directly related to major neuropsychiatric diseases, such as depression, Parkinson's disease and others (Kiss JP Role of nitric oxide in the regulation of monoaminergic neurotransmission. Brain Res. Bull., 2000, Aug., 52 (6): 459-66).

Elevated levels of monoamine oxidase, oxidative stress, excitotoxicity and excessive NO synthesis are characteristic of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, stroke and others (Mandel S., Grunblatt E., Riederer P., Gerlach M., Levites Y., Youdim MB Neuroprotective strategies in Parkinson's disease: an update on progress. CNS Drugs, 2003, 17 (10): 729-62).

The development of addictions and psychoses is based on behavioral sensitization mediated by the activation of D1 dopamine receptors, as well as NMDA receptors, followed by increased NOC activity (Ujike H. Advanced findings on the molecular mechanisms for behavioral sensitization to psychostimulants. Nippon Yakurigaku Zasshi., 2001, Jan ., 117 (1): 5-12).

Excessive release of NO from blood vessels, perivascular nerve endings, or the brain is considered the molecular mechanism triggering spontaneous migraine pain (Olesen J., Jansen-Olesen I. Nitric oxide mechanisms in migraine. Pathol. Biol., Paris, 2000, Sep. 48 (7): 648-57).

Dopamine (hereinafter DA) is the most important neurotransmitter and neuromodulator, which plays an important role in the body. In the central nervous system, DA is involved in the control of movement, cognitive functions, emotionality, neuroendocrine secretion, and retinal cell function. On the periphery, DA is involved in the regulation of homeostasis, vascular tone and hormone secretion. In the central nervous system, dopamine receptors are widely represented in various regions of the brain (Missale C., Nash SR, Robinson SW, Jaber M., Caron MG Dopamine receptors: from structure to function. Physiol. Rev., 1998, Jan. 78 (1 ): 189-225). The various physiological functions of DA are mediated by at least five DA: D1-D5 families of receptors. Dysfunction of these receptors is observed in various disorders and diseases of the central nervous system, in particular Parkinson's disease (Zawilska J.B. Dopamine receptors-structure, characterization and function. Postepy, Hig. Med. Dosw., 2003, 57 (3): 293-322).

Dopamine signaling in certain parts of the brain is a key element in the development of drug dependence (Fagen ZM, Mansvelder HD, Keath JR, Mc.Gehee DS Short - and long-term modulation of synaptic inputs to brain reward areas by nicotine. Ann., NY Acad. Sci ., 2003, Nov., 1003: 185-95). Interaction with DA receptors underlies the acute effects of amphetamine and cocaine (Ujike H. Molecular biology of drug dependence and behavioral sensitization. Seishin Shinkeigaku Zasshi., 2002, 104 (11): 1055-68; Wolf ME, Mangiavacchi S., Sun X . Mechanisms by which dopamine receptors may influence synaptic plasticity. Ann. NY Acad. Sci., 2003, Nov. 1003: 241-9). Dopamine agonists mediate dependence on alcohol, nicotine and stimulants (Kosten T.R., George T.P., Kosten T.A. The potential of dopamine agonists in drug addiction. Expert Opin. Investig. Drugs, 2002, Apr. 11 (4): 491-9).

There is a hypothesis that dopaminergic system disorders underlie schizophrenia (Pearlson G. D., Neurobiology of schizophrenia. Ann. Neurol., 2000, Oct., 48 (4): 556-66). The dopamine hypothesis of schizophrenia postulates an imbalance in the cortical / subcortical dopamine system and impaired functioning of the D1 dopamine receptors (Abi-Dargham A., Moore H. Prefrontal DA transmission at D1 receptors and the pathology of schizophrenia. Neuroscientist, 2003, Oct., 9 (5): 404-16). In addition to schizophrenia, the etiology and other psychotic diseases are associated with impaired regulation of the dopamine system of the brain (Conley RR, Kelly DL Current status of antipsychotic treatment. Curr. Drug Target CNS. Neurol. Disord., 2002, Apr., 1 (2): 123- 8).

Dopamine mechanisms mediate the etiology and manifestations of anxiety (Taylor DP, Riblet LA, Stanton HC, Eison AS, Eison MS, Temple DL Jr., Dopamine and antianxiety activity. Pharmacol. Biochem. Behav., 1982, 17, Suppl. 1: 25- 35), and many antipsychotic drugs modulate the activity of D2 dopamine receptors (Kapur S., Mamo D. Half a century of antipsychotics and still a central role for dopamine D2 receptors. Prog. Neuropsychopharmacol. Biol. Psychiatry, 2003, Oct., 27 (7): 1081-90).

Thus, in the development of many pathologies, disorders of the nitrergic system are directly associated with disorders of the dopaminergic system. These pathologies include chemical addiction - substance abuse disorders such as drug, alcohol and nicotine addiction, sleep disorders, sexual disorders including sexual dysfunctions, gastrointestinal disorders, psychoses, affective disorders, inorganic psychoses, personality disorders, psychiatric mood disorders, schizophrenia and schizoaffective disorders, polydipsia, bipolar disorders, dysphoric mania, anxiety and related diseases, obesity bacterial infections of the central nervous system, such as meningitis, learning disabilities, memory impairments, Parkinson's disease, depression, extrapyramidal side effects of antipsychotics, hypothalamic-pituitary disorders, vascular and cardiovascular diseases, dystonia, dyskinesias, hyperkinesia, dementia, ischemia, motor disorders, hypertension, and diseases caused by a hyperactive immune system, such as allergies and inflammations, in mammals, including humans.

The development of drugs for the treatment of disorders caused by impaired function of the nitrergic and dopaminergic systems by normalizing these systems is a very urgent task.

Known tricyclic amines with central dopaminergic activity (US, 4612316, A), as well as dihydropyridine carboxamides, dihydroquinoline and isoquinoline carboxamides (US, 4727079, A), new derivatives of aminotriazoles and aminooxazoles (US, 4904676, A) with specific brain energies activity.

The use of benzothiopyranopyridinones for the induction of antipsychotic, antidepressant and antidopaminergic activity in warm-blooded animals is known (US, 4547507, A).

The use of phenoldopam 4 ', 8-bis-hydrosulfate and its salts is known as preparations with prodopaminergic activity (US, 4600714, A).

The use of (1,2,5,6-tetrahydro-1-alkyl-3-pyridinyl) -2-thiazolamines and 4- (hexahydro-1-alkyl-3-pyridinyl) -2-thiazolamines with suggested antipsychotic activity for the treatment of psychoses is proposed , high blood pressure, Parkinson's disease, hyperprolactinemia, sexual dysfunctions and acromegaly (US, 46508054, A).

Substituted 1- (alkoxyphenyl) piperazines, partial dopamine agonists that are proposed to be used to treat dopamine regulation disorders and treat Parkinson's disease, schizophrenia and drug addiction (US, 5281594, A), are known.

Derivatives of benzimidazolone with central dopaminergic activity (US 5889010, A; US, 5883094, A), as well as N, N'-disubstituted derivatives of benzimidazolone (US, 6521623, A), which can possibly be used to treat a wide range of diseases, are proposed. nervous system.

In the mentioned analogs, only data on the binding of the corresponding compounds to dopamine receptors are presented, but in no case was the presence of specific therapeutic activity in animal models or in clinical trials.

Disclosure of invention

The aim of the present invention is to provide a drug for the correction of disorders of the nitrergic and dopaminergic systems and disorders caused by disorders of these systems, in particular diseases of the nervous and cardiovascular systems, including disorders caused by substance abuse, impaired cognitive function and psychophysiological state.

When creating the present invention, the task was to identify biologically active compounds with the ability to normalize the work of both nitrrhagic and dopaminergic systems by regulating the level of NO in the cell by correcting the activity of various isoforms of NO synthase, as well as by binding excessively formed active forms of nitrogen, in particular peroxynitrite or an NO radical.

As biologically active compounds possessing the properties necessary for solving this problem, cyclic bioisosteres of derivatives of the purine system having the general structural formula were considered

R ¹ = —H, —NH ₂ , —Br, —Cl, —OH, —COOH,

B = -N =, -CH =, Z = -CH =, -N =,

A = -N = with B = -N =, Z = -CH-,

A = -CH = at B = -N =, Z = -CH-,

A = -CH = at B = -N =, Z = -N =,

A = -CH = at B = -CH =, Z = -CH =,

A = -CH = at B = -CH =, Z = -N =,

and their pharmacologically acceptable salts, which have a normalizing effect on intracellular processes, in particular on the nitrergic mechanisms of cells (PCT / RU 03/00346). The authors suggested that condensed pyridazinedione systems have specific neurotropic activity, and the positive effect is associated with the normalization of the disturbed functions of the nitrergic and dopaminergic systems by these compounds.

The mechanisms of the effect of cyclic bioisosteres of purine derivatives on the nitrergic system can include a change in the pH in the cell, affecting the activity of nitric oxide synthase, direct interaction of these compounds with nitric oxide and peroxynitrite in the cell and outside the cell, which affects both the nitrergic system as a whole and and on the free radical homeostasis of a biological object. These compounds can also differentially modulate the activity of various isoforms of nitric oxide synthase, which to a large extent provides a change in the functional state of cells, organs, tissues, and, ultimately, the whole organism. The effect of the compounds of the invention on the dopamine system can be achieved by changing the distribution of electron density in the protein molecules of dopamine receptors or by changing the properties of the membranes surrounding the receptors. The additional effect of these compounds on other types of receptors, for example, adenosine, can indirectly modulate the activity of dopamine receptors.

Derivatives of pyrido [2,3-d] -6H-pyridazin-5,8-dione, a cyclic bioisoster of derivatives of the purine system, in which a pyridine ring is fused to a pyridazinedione ring having the general formula

атом Li, Na, К,

atom Li, Na, K,

R ¹ = —H, —NH ₂ , —Br, —OH, —COOH,

in particular:

7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (1),

4-amino-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (2),

3-bromo-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (3),

4-hydroxy-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazine-5,8-dione disodium salt (4),

3-carboxy-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazine-5,8-dione disodium salt (5),

pyrido [2,3-d] -6H-pyridazin-5,8-dione lithium salt (6),

pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (7),

pyrido [2,3-d] -6H-pyridazin-5,8-dione potassium salt (8).

Derivatives of benzo [d] -3H-pyridazine-1,4-dione, a cyclic bioisoster of derivatives of the purine system, in which the benzene ring is condensed with the pyridazinedione ring, having the general formula

атом Li, Na, К,

atom Li, Na, K,

R ¹ = —H, —NH ₂ , —Cl, —OH, —COOH,

in particular:

2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (9),

5-amino-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (10),

6-amino-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (11),

5-chloro-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (12),

5-hydroxy-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione disodium salt (13),

5-amino-benzo [d] -3H-pyridazin-1,4-dione lithium salt (14),

5-amino-benzo [d] -3H-pyridazin-1,4-dione sodium salt (15),

6-amino-benzo [d] -3H-pyridazin-1,4-dione potassium salt (16),

5-hydroxy-benzo [d] -3H-pyridazin-1,4-dione disodium salt (17),

6-carboxy-benzo [d] -3H-pyridazine-1,4-dione disodium salt (18).

Derivatives of pyrazino [2,3-d] -6H-pyridazine-5,8-dione, a cyclic bioisoster of derivatives of the purine system, in which the pyrazine ring is fused to the pyridazinedione one having the general formula

атом Li, Na, К,

atom Li, Na, K,

R ¹ = —H, —NH ₂ , —Br, —OH, —COOH,

in particular:

7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (19),

2-amino-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (20),

3-amino-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (21),

3-bromo-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (22),

2-hydroxy-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazine-5,8-dione disodium salt (23),

2-carboxy-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazine-5,8-dione disodium salt (24),

pyrazino [2,3-d] -6H-pyridazin-5,8-dione lithium salt (25),

pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (26),

3-bromo-pyrazino [2,3-d] -6H-pyridazin-5,8-dione potassium salt (27),

2-amino-pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (28).

Derivatives of pyrimido [4,5-d] -6H-pyridazine-5,8-dione, a cyclic bioisoster of derivatives of the purine system in which the pyrimidine ring is fused to the pyridazinedione ring having the general formula

атом Li, Na, К,

atom Li, Na, K,

R ¹ = —H, —NH ₂ , —Br, —OH, —COOH,

in particular:

7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazine-5,8-dione sodium salt (29),

2-amino-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (30),

4-amino-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (31),

2-bromo-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (32),

4-hydroxy-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (33),

4-carboxy-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazine-5,8-dione sodium salt (34),

pyrimido [4,5-d] -6H-pyridazin-5,8-dione lithium salt (35),

2-amino-pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (36),

4-bromo-pyrimido [4,5-d] -6H-pyridazine -5,8-dione potassium salt (37).

Compounds 1-8, which are derivatives of pyrido [2,3-d] -6H-pyridazin-5,8-dione, were obtained by condensation of ortho-dicarboxy substituted pyridines with hydrazine hydrate in acetic acid (Taguchi Hiroshi. A new fluorometric assay method for guinolinic acid. Analitic Biochemistry, 1983, 131 (1), p. 194-197).

Compounds 9-18, which are derivatives of benzo [d] -3H-pyridazin-1,4-dione (phthalazinedione), were obtained by condensation of orthophthalic acid with hydrazine hydrate in acetic acid (Huntress EH, Stanley LN, Parker AS The preparation of 3 -Aminophtalhydrazide for use in the Demonstration of Chemiluminescence, J, Am. Chem. Soc., 1994, v. 56, p. 241-242).

Compounds 19-28, which are derivatives of pyrazino [2,3-d] -6H-pyridazine-5,8-dione, were obtained by condensation of ortho-dicarboxy substituted pyrazines with hydrazine hydrate in acetic acid (Zyczynska - Baloniak I., Czajka R., Zinkowska E ., Synthesis of Derivatives of 4-Hydroxypyrazine- [2,3-d] pyridazine-1-one. Polish Journal of Chemistry. 1978, v.52, p.2461-2465; Kormendy K., Ruff P. Pyridazines condensed with a Heteroring. Sh., Acta Chimika Hungarika. 1990, 127 (2), p. 253-262). Compounds 29-37, which are derivatives of pyrimido [4,5-d] -6H-pyridazine-5,8-dione, were obtained by condensation of ortho-dicarboxy substituted pyrimidines with hydrazine hydrate and acetic acid (Yurugi S., Hieda M. Studies on the synthesis of N-Heterocyclic Compounds. Chemistry, Pharmaceutic Bull., 1972, v. 20 (7), p. 1522-1527, ibid., p. 1513-1521).

The problem was solved by identifying the biological activity of the above cyclic bioisosteres of the purine system and their pharmacologically acceptable salts on the nitrergic and dopaminergic mechanisms of body cells having disorders of such systems.

The invention is further illustrated by the following non-limiting examples, illustrating in vivo experiments on models of various pathologies, the effect of the use of these compounds in disorders caused by a violation of the nitrergic and dopaminergic systems of the animal body.

1. The effect of the compounds of the invention on disorders caused by morphine withdrawal.

We studied the effects of compounds No. 1, 4, 6, 10, 11, 18, 21, 24, 26, 32, 35, 37 according to the invention on behavioral indicators, as well as on the nitrergic system of the brain of animals, in which, according to a known model, recognized also a model of human heroin withdrawal, a state of physical dependence on morphine - morphine withdrawal - was modeled.

Physical dependence on morphine was modeled on 27 groups of male Wistar rats weighing 250-350 g 6 months of age:

- animals of group No. 1 (n = 7) were not injected with morphine, and they were a control,

- animals of group No. 2 (n = 7) - were injected morphine hydrochloride to achieve a pronounced withdrawal syndrome,

- animals of groups No. 3-14 (n = 7) were administered one of the compounds of the pyridopyridazinedione series - 1, or 4, or 6 according to the invention (groups No. 3, 4, 5), or one of the compounds of the benzopyridazinedione series - 10, or 11, or 18 according to the invention (groups No. 6, 7, 8), or one of the compounds of the pyrazinopyridazinedione series - 21, or 24, or 26 according to the invention (groups No. 9, 10, 11), or one of the compounds of the pyrimidinopyridazinedione series - 32, or 35, or 37 (groups 12, 13, 14),

- animals of groups No. 15-26 (n = 7) were injected with morphine hydrochloride to create a pronounced withdrawal syndrome and then: in each group, one of the compounds according to the invention according to a scheme similar to groups No. 3-14.

Injections of morphine hydrochloride were carried out intraperitoneally according to a modified scheme: for 6 days twice a day (at 10:00 and 20:00) in increasing doses from 10-100 mg / kg: 1 day - 10 mg / kg, 2 day - 20 mg / kg, 3 days - 40 mg / kg, 4 days - 60 mg / kg, 5 days - 80 mg / kg, 6 days - 100 mg / kg. Injections of the compounds according to the invention were carried out three times intramuscularly at a dose of 20 mg / kg during the day following the last dose of morphine. (Dum J, Blasig J, Herz A: Buprenorphine: demonstration of physical dependence liability. Eur. J. Pharmacol., 1981, V.70, p. 293-300 .; Rahman S., Ali Khan R., Kumar A. Experimental study of the morphine de-addiction properties of Delphinium denudatum Wall, BMC Complement Altern. Med., 2002, V.29, p. 1-6). 36 hours after the last injection, the motor and vegetative signs of animal behavior were evaluated in groups by signs that are specific signs of withdrawal in the open field test (arena with a diameter of 120 cm and a wall height of 40 cm).

The severity of withdrawal symptoms was assessed for 5 minutes using a number of motor signs specific to the withdrawal syndrome: shaking as a “wet dog”, cramps, chewing, gritting of teeth, shaking with front paws, and autonomic signs: diarrhea, ptosis, rhinorrhea, piloerection, dyspnea, squeaking when touching, aggressiveness (Blasig J., Herz A., Reinhold K., Zieglgansberger S. Development of physical dependence on morphine in respect to time and dosage and quantification of the precipitated withdrawal syndrome in rats. Psychopharmacologia, Berlin, 1973, V .33, p.19-38; Rahman S., Alt Khan R., Kumar A. Experimental study of the morphine de-addiction properties of Delphinium denudatum Wall, BMC Complement Altern. Med., 2002, V.29, p. 1-6.). The observed signs were recorded quantitatively (if possible) with further assignment to each attribute of the index (depending on the specificity of the attribute) and calculation of the sum of points. The severity of withdrawal symptoms was presented as the sum of the points. The results were processed using nonparametric statistical analysis using the Mann-Whitney test.

Figure 1 shows a diagram of the effect of morphine and compounds according to the invention on the behavioral reactions of the studied animals, on which the effect is estimated by the sum of M points averaged for a number of compounds based on the indicated indices of withdrawal symptoms for animal groups No. 1-27.

Table 1 shows the averaged relative data.

Table 1
The effect of the compounds of the invention on the development of morphine withdrawal Specific symptoms of withdrawal Index Significant differences (criterion Chi _Greek - square) in the manifestation of signs of withdrawal Control - Morphine Morphine- (Morphine + compound 1, or 4, or 6) Morphine- (Morphine + Compound 10, or 11,
or 18) Morphine- (morphine + compound 21, or 24,
or 26) Morphine- (morphine + compound 32, or 35, or 37) shaking the wet dog 2 0,0002 1,0000 0.7562 0.7821 1,0000 writhes 2 0,0507 0.7821 0.7546 0.3456 0.5578 chewing 2 0,0308 0.1888 0.1032 0.1975 0.1888 gritting teeth 2 0.1266 0.7144 0.8652 0.2994 0.7003 forefoot shaking 2 0.0180 0.7821 0.8217 0.6745 0.9321 squeaking when touching one 0.0053 0.1888 0.1342 0.1968, 0.2035 diarrhea one 0.0075 0.0046 0.0032 0.0067 0.0001 ptosis 2 0.0053 0,0201 0,0320 0.0105 0.0232 rhinorrhea 3 0.2994 0,3017 0.4032 0.2131 0.2935 piloerection 2 0.5770 0.1847 0.2567 0.2567 0.2345 dyspnea 2 0.0053 0,0722 0.0834 0,0567 0,0685 aggressiveness one 0.5770 0.8327 0.8456 0.7921 0.8456

From table 1 it is seen that of the signs characterizing the development of withdrawal symptoms, the compounds according to the invention most effectively stopped diarrhea, ptosis and dyspnea. At the same time, they also affected the convulsive activity caused by the abolition of morphine.

Thus, it was possible to identify certain components of morphine withdrawal that are sensitive to correction by the compounds according to the invention.

2. The effect of the compounds of the invention on nitrergic parameters in the brain.

2.1. Obtaining material for biochemical studies

After the above experiments, the brain was removed from the rats and immediately placed in an ice-cold 0.9% sodium chloride solution. Following cooling, the following structures were isolated from the brain: cerebral cortex, hippocampus, midbrain, striatum, brain stem, hypothalamus and cerebellum. The isolated tissue was homogenized in a Potter S homogenizer for 3 min at 1500 rpm in 4-5 volumes of 20 mM HEPES (pH 7.5) at 4 ° C. The supernatants were centrifuged for 30 min at 11000 g at 4 ° C, and a part of the obtained supernatants were selected for determination of nitrates and nitrites (NO _x ^- ), and the remaining part was added with cooled 20 mM HEPES (pH 7.5) containing 0.5 mM ethylenediaminetetraacetate (EDTA), 1 mM dithiothreitol (DDT), 1 mM phenylmethylsulfonyl fluoride (FMSF), aprotinin and leipeptin at a concentration of 5 μg / ml, and were used to determine the activity of the enzyme nitric oxide synthase (NOC).

2.2. Determination of nitrates / nitrites (NO _x ^- )

To assess the intensity of rat nitric oxide metabolism, stable nitrite oxide metabolites - nitrites and nitrates (NO _x ^- ) were quantified by fluorimetric method according to the fluorescence intensity of naphtotriazole, a reaction product of 2,3-diaminonaphthalene (DAN) and nitrite in an acidic medium (Misko T.K. ., Schilling RJ, Salvemini D. et al. A fluorometric assay for the measurement of nitrite in biological samples. Anal. Biochem., 1993, V.214, p.11-16) with modifications (Lei B., Adachi N. , Nagaro T., Arai T. Measurement of total nitric oxide metabolite (NO (x) (-)) levels in vivo. Brain, Res. Protoc., 1999, V.4, p. 415-419).

The brain supernatants deproteinized at 100 ° C were placed in a nitrite regenerating system containing 0.125 U / ml nitrate reductase, 25 μM NADFN and 25 μM FAD prepared in 20 mM Tris-HCl buffer pH 7.6 and incubated for 30 min at 37 ° C. For the oxidation of NADFN, the lactate dehydrogenase (LDG) / pyruvate system was used. Then, 316.0 μM DAN solution in 0.62 M HCl was added and incubated for 10 min in the dark. To stabilize the fluorescence of the resulting naphtotriazole, 280 mM NaOH was added. The fluorescence intensity was measured on a Hitachi F-3000 spectrofluorimeter at an excitation wavelength of 365 nm and an emission of 405 nm. To calculate the concentration of NO _x ^- in the brain, a standard solution of sodium nitrate was used. The concentration of NO _x ^{- was} expressed in nmol / mg protein.

2.3. Determination of nitric oxide synthase activity

The activity of nitric oxide synthase (NOC) was determined by radiometric method according to the rate of accumulation of L-citrulline in the oxidation reaction of [ ³ H] L-arginine catalyzed by NOC (Bredt and Snyder. Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum. Proc. Natl. Acad. Sci., USA, 1989, V. 86, p. 9030-9033). The formation of L-citrulline in this reaction is equivalent to nitric oxide biosynthesis.

The reaction was initiated by adding a brain supernatant to a reaction medium containing 2 μCi / ml [ ³ H] L-arginine, 20 mM HEPES (pH 7.4), 0.2 mM CaCl ₂ 5 μM FAD, 5 μM FMN, 1 mM NADFN, 50 μM VN ₄ in the study of brain supernatants. After 15-60 minutes of incubation at 37 ° C, a Dowex 50WX8-400 suspension (Na ⁺ form) was added to the samples, which sorb unreacted [ ³ H] L-arginine, but not [ ³ H] L-citrulline. After sorption, the radioactivity of the samples was determined on a SL-4000 scintillation counter (Intertechnique). The activity of Ca ²⁺ -dependent and Ca ²⁺ -independent NOC isoforms was determined by the difference in the rates of [ ³ H] L-citrulline formation in three parallel samples containing 2 mM EDTA (Ca ²⁺ chelator), 2 mM EDTA + 2 mM L-NAME (inhibitor of all NOC isoforms) and without inhibitors. Enzyme activity was expressed in pmol of [ ³ H] L-citrulline accumulated over 1 min per mg of protein in the supernatant.

2.4. Protein quantification

Protein content in the samples was determined by the Bradford method (Bradford M. M. A rapid and sensitive method for quantitation of microgram quantities of protein using the principle of protein binding. Anal. Biochem., 1976, V.72, p. 248-254) using Coomassie Blue Dye. Statistical analysis was carried out by methods adequate for a particular sample. Data are presented as mean ± error of the mean.

table 2
The effect of morphine and compounds according to the invention on the performance of the nitrergic system in the brain Indicator in the brain Group No. 1 (control) Group No. 2 (morphine) Groups No. 3-14 (Connection from the group 1, 4, 6, 10, 11, 18, 21, 24, 26, 32, 35, 37) Groups No. 15-17 (Morphine + compound 1, or 4, or 6 Groups No. 18-20 (Morphine + compound 10, or 11, or 18) Groups No. 21-23 (Morphine + compound 21, or 24, or 26) Groups No. 24-27 (Morphine + compound 32, or 35, or 37) Nitrites, nmol / mg protein: cerebral cortex 4.63 ± 0.29 5.31 ± 0.29 4.31 ± 0.37 4.72 ± 0.37 4.93 ± 0.39 4.62 ± 0.27 4.84 ± 0.31 cerebellum 6.37 ± 0.64 6.07 ± 0.42 6.66 ± 0.63 5.46 ± 0.44 5.85 ± 0.47 5.76 ± 0.49 5.93 ± 0.54 trunk 8.24 ± 1.02 7.63 ± 0.96 6.31 ± 0.58 6.57 ± 0.72 6.99 ± 0.82 7.07 ± 0.85 6.87 ± 0.81 striatum 7.25 ± 0.66 4.92 ± 0.43 6.11 ± 0.29 3.57 ± 0.42 3.57 ± 0.42 3.57 ± 0.42 3.57 ± 0.42 hippocampus 4.39 ± 0.23 6.45 ± 0.69 6.47 ± 0.72 5.44 ± 0.43 5.24 ± 0.49 5.27 ± 0.41 5.31 ± 0.33 midbrain 5.66 ± 0.19 9.41 ± 1.20 6.65 ± 0.70 5.50 ± 0.48 5.75 ± 0.39 5.99 ± 0.58 5.95 ± 0.52 hypothalamus 6.57 ± 0.50 4.62 ± 0.71 6.77 ± 0.83 6.57 ± 0.88 6.91 ± 0.58 6.87 ± 0.75 6.77 ± 0.46 NOC activity, pmol / min / mg protein: striatum 1.73 ± 0.07 1.19 ± 0.14 1.26 ± 0.08 1.24 ± 0.08 1.29 ± 0.09 1.21 ± 0.10 1.34 ± 0.09 midbrain 2.18 ± 0.09 3.08 ± 0.09 2.60 ± 0.29 1.83 ± 0.23 1.99 ± 0.29 2.03 ± 0.31 1.89 ± 0.27 hippocampus 2.55 ± 0.12 3.10 ± 0.20 3.09 ± 0.16 2.70 ± 0.14 2.51 ± 0.19 2.33 ± 0.11 2.65 ± 0.15 hypothalamus 5.37 ± 0.20 3.42 ± 0.54 5.45 ± 0.24 5.26 ± 0.28 5.36 ± 0.33 5.47 ± 0.32 5.56 ± 0.34

As can be seen from the results shown in Table 2, in the brain, morphine had a specific effect on the accumulation of nitrites and NOC activity: in the striatum and hypothalamus, a decrease in nitrergic indices was observed, and in the middle brain and hippocampus their increase (P <0.05; P = 0.1 for nitrites in the hypothalamus, T-test). The compounds of the invention stopped morphine-disrupted NOC activity in the hypothalamus, midbrain and hippocampus.

findings

Thus, with the withdrawal syndrome of morphine, the compounds according to the invention, administered three times at a dose of 3 × 20 mg / kg intramuscularly, have the following effects:

- stop the development of withdrawal symptoms, primarily such components of behavioral disorders as diarrhea, ptosis and dyspnea,

- improve the psychophysiological state due to the antidepressant effect of inhibition of withdrawal symptoms,

- have a differentiated and specific effect on various isoforms of nitric oxide synthase, correcting impaired nitrergic mechanisms in the brain.

These data indicate the promise of the use of the compounds according to the invention for the treatment of disorders caused by substance abuse, in particular drugs.

3. The effect of the compounds of the invention on sexual dysfunction associated with dopaminergic system disorders.

One of the mechanisms regulating the sexual function of mammals is associated with the functioning of the dopaminergic system of the brain. A low-dose non-selective D1 / D2 agonist of apomorphine is known to cause penile erection in rodents (Giuliano F., Allard J. Dopamine and male sexual function. Eur. UroL, 2001, 40 (6), 601-608; Giuliano F. , Allard J., Rampin O. et. Al. Pro-erectile effect of systemic apomorphine: existence of a spinal site action. J. UroL, 2002, 167 (1), 402-406; Brien SE, Smallegange C., Gofton WT, et. Al. Development of a rat model of sexual performance anxiety: effect of behavioral and pharmacological hyperadrenergic stimulation on apomorphine-induced erections. Int. J. Impot. Res., 2002, 14 (2), 107-115.) . In this regard, a study was conducted of the effect of the compounds according to the invention on apomorphine-dependent erection in rats.

In the experiment, 9 groups of adult male Wistar rats weighing 350-450 g, kept under normal light conditions, were used. The experiment involved rats with a normal (non-inverted) light cycle. To study the effect of the compounds on sexual function, low-grade animals with one erection were selected. Control group No. 1 (n = 7) consisted of animals not injected with the compounds of the invention. For animals of groups No. 2-9 (n = 7), one of the compounds according to the invention, selected from the group of compounds 2, 8, 9, 15, 19, 25, 31, 36, was administered intraperitoneally with a course of: 5 injections at a dose of 10 mg / kg with an interval between injections of 48 hours. Apomorphine was dissolved in a 0.1% aqueous solution of ascorbic acid and then was administered to everyone subcutaneously at a dose of 0.1 mg / kg 24-28 hours after the last injection of the compounds of the invention. Registration of sexual activity was carried out individually for each animal immediately after administration of apomorphine; registration time 20 min. Registered: the time of onset of the first erection, the time intervals between erections, the number of erections for the entire observation period.

For statistical processing of the results, the T-test and the Chi-square test were used. The results are presented in table 3 in the form of mean values ± error of the mean.

Table 3 The effect of the compounds of the invention on sexual dysfunction in rats Group (experimental conditions) Latency period of the first erection, min The number of erections for the entire observation period The interval between the 1st and 2nd erection, min The interval between the 2nd and 3rd erection, min No. 1 (Control + apomorphine) 6.63 ± 0.99 2.00 ± 0.37 4.60 ± 0.53 5.90 ± 0.87 No. 2-3 (compound 2 or 8 + apomorphine) 5.21 ± 0.30 P = 0.1 3.13 ± 0.35 P <0.05 3.94 ± 0.60 5.52 ± 1.14 No. 4-5 (compound 9 or 15 + apomorphine) 5.09 ± 0.27 P <0.1 3.24 ± 0.39 P <0.05 3.87 ± 0.59 5.19 ± 0.78 No. 6-7 (compound 19 or 25 + apomorphine) 5.24 ± 0.19 P <0.1 3.29 ± 0.27 P <0.05 3.75 ± 0.64 5.12 ± 0.87 No. 8-9 (compound 31 or 36 + apomorphine) 5.12 ± 0.24 P <0.1 3.32 ± 0.31 P <0.05 3.34 ± 0.69 5.02 ± 1.01

From the results of the studies presented in table 3, it can be seen that the compounds according to the invention significantly increased the number of erections in animals (more than 1.5 times) and showed a statistically significant tendency to decrease the latency of the first erection by 1.3 times.

findings

Thus, a beneficial effect of the compounds according to the invention on the sexual function of rats was revealed.

Since the model used involves certain cerebral mechanisms, it can be considered that the mechanism of action of the compounds according to the invention is associated with its effect on the dopaminergic system of the brain, in particular, elimination of dopamine D1 / D2 dysfunction. From the results obtained, it is obvious that the compounds according to the invention can also be used to correct numerous pathologies of the nervous system associated with dysfunction of the dopaminergic system.

4. The effect of the compounds according to the invention on the cognitive functions and psychophysiological state of animals

A screening study of the effects of the compounds according to the invention on the learning and memory processes, as well as on the psychophysiological state of rats, was carried out.

4.1. Experimental technique

Wistar rats weighing 220-300 g were used in the study. In each test, control No. 1 (n = 10) and experimental groups No. 2-9 (n = 10) were used.

Animals of groups No. 2-9 were administered one of the compounds from the group of compounds No. 2, 7, 11, 17, 20, 28, 29, 35 according to the invention, intramuscularly, once at a dose of 10 mg / kg The preparations were dissolved in neutralized water. To study the effect of the compounds according to the invention, widely used tests were used to assess learning and memory: active avoidance, passive avoidance, and the psychophysiological state of animals: anxiety, aggressiveness, depression. These tests are usually used for screening nootropic and psychopharmacological drugs, as well as various effects on the cognitive and emotional sphere.

4.1.1. Learning and memory assessment

A conditional passive avoidance reaction (passive avoidance reaction) was developed on the basis of a single electrodermal reinforcement in an installation consisting of two chambers - large (illuminated 25 × 25 × 25 cm) and small (dark 17.5 × 14 × 14 cm) with electrified floors communicating with each other a rectangular hole of 7 × 10 cm. During training, the rat was placed for 3 min in the middle of the illuminated chamber with its tail to the hole in the dark. The animal examined the camera, found a hole and goes into the dark compartment. Due to its biological characteristics, the rat prefers to be in a dark room. The latent period of the first entry into the dark chamber (T1) was recorded, then immediately after the entry, the rat was landed from the dark chamber. After 30 min, the manipulations were repeated and the latent period of the second entry into the dark chamber (T2) was recorded. After another 30 min, the latent period of the third entry into the dark chamber (T3) was recorded and the opening was closed with a door (the animal remained inside). Then, alternating electric current (50 Hz, 80 V) was applied to the electrode floor for 5 s, after which the rat was planted in a home cage. Testing of the retention of the acquired reaction was carried out after 24 hours, 7 and 14 days. For this, the rat was placed in the apparatus for 3 min and the latent period of entry into the dark chamber was recorded. The learning criterion is a latent transition period of at least 180 s. A longer residence time of the animal in the illuminated part of the installation or refusal to go into the dark chamber is considered as preserving the acquired reaction.

4.1.2. Assessment of psychophysiological status

a) Anxiety.

Anxiety is considered to be the behavior associated with the prevalence of fear motivation. Among the various types of anxiety, phobias are most common, especially simple phobia and agoraphobia (fear of open space). Testing rats in an elevated cruciform labyrinth is one of the most widely used methods for studying rodent anxiety. The labyrinth we use is made of wood coated with green plastic. The labyrinth consists of two open sleeves (50 × 15 cm), located opposite each other, and two closed sleeves of the same size, having walls 20 cm high along the long sides of the sleeve. The labyrinth is located at a height of 70 cm from the floor surface. Testing was carried out in a soundproofed room. The rat was placed in the central part of the installation and for 10 min the number of entries into the open and closed arms of the maze and the time spent in the open and closed arms were visually recorded. Each animal was tested once.

b) Depressiveness

Depressive states include affective disorders of mental activity, characterized by emotional indifference, feelings of unhappiness, thoughts of death, suicidal manifestations, changes in psychomotor behavior, impaired cognitive functions (primarily inability to concentrate, memory impairment). One of the most appropriate and widely used methods for detecting conditions of similar depression in animals is the forced swimming method (Porsolt swimming test). During the test, the animal was placed in a vessel with water without the ability to independently avoid this stressful situation. The indicators of depression in the test conditions were the number and duration of periods of passive swimming (retention on the surface of the water without visible movements of the paws). This type of reaction reflects a state of behavioral "despair." The experiments were carried out in a circular pool with a diameter of 40 cm and a height of 50 cm. The pool was filled with water t = 22 ° C to a height of 30 cm. The rat was released into the pool and the following indicators of behavior were recorded: the duration of the first period of active swimming (swimming with random movement of limbs, climbing ), the duration of passive swimming (periods of hovering without movement), the duration of active swimming. Testing was carried out for 5 min, once. Statistical analysis was performed using appropriate statistical tests in each case.

The results of the experiments are presented in Figa, 2b, 3a, 3b, 3c, 4a, 4b, where:

- Fig.2a, 2b shows the effect of the compounds according to the invention on learning in a conditioned passive avoidance reaction for animal groups No. 1-9: Fig.2a shows the latent period t _{l of} entering the dark compartment after electroshock, Fig.2b shows the number N trained animals, while the values in the T0 region are the initial values of N before the current is applied, in the T1, T7, T14 regions after 1, 7 and 14 days, respectively.

- figa, 3b, 3c shows the effect of the compounds according to the invention on the anxiety of rats of groups No. 1-9 in the elevated cross-shaped labyrinth: Fig. 3a shows the number of entries N ₁ into the open (region O) and closed (region C) sleeve, on fig.3b - time t spent in an open sleeve, on figs - the number of animals N ₂ that did not go into the open sleeve,

- figa, 4b shows the effect of the compounds according to the invention on the depressiveness of animal groups No. 1 to 9 in the Porsolt forced swimming test: Fig. 4a shows the swimming time t _s before the first freeze (region A) and the active swimming time t _s ( region B), FIG. 4b is the number N _{3 of} hangs.

The results are presented as mean ± error of the mean.

Visually, there were no violations of the behavior or appearance of rats after administration of the drug.

In the passive avoidance test (Fig. 2a, 2b), the compounds according to the invention significantly (P <0.05, T-test) increased the latent period t _{l of} entering the dark compartment one day after the "electric shock (Fig. 2a), which indicates a more effective 2b shows that the number N of trained animals is twice as large in the groups receiving the compounds according to the invention (statistically significant trend, P <0.07, Chi-square test). N trained animals and the latency period t _l remained above in g uppah №2-9, received the inventive compounds than in the control group №1, after 1 and 2 weeks significance of differences became significant.

Anxiety test in the elevated cross-shaped labyrinth (Fig. 3a, 3b, 3c) revealed greater research activity (and, therefore, less anxiety) in the groups receiving the compounds according to the invention — the number N _{1 of} entries in the open and closed sleeve was significantly higher (P <0.05, T-test) than in the control group (Fig.3a), while the time t spent in the open sleeve was 2.2 times longer in the groups receiving the compounds according to the invention (Fig.3b), although from Due to the variability of the data, the differences were only at the level of the trend (P = 0.01, T-test). By the number N _{2 of} animals that did not go into the open sleeve (these are animals with the most pronounced anxiety), the control group exceeded the experimental group by 6 times (6 out of 10 and 1 out of 10, respectively, P <0.02, Chi-square test) (Fig.3c )

Thus, the compounds of the invention significantly reduced anxiety when tested in an elevated cruciform maze.

The depression test (forced swimming) (Figs. 4a, 4b) did not reveal a significant effect of the compounds according to the invention either in the total active swimming time t _s (Fig. 4a) or in the number N _{3 of} hang (Fig. 4b). Nevertheless, rats of groups No. 2-9 receiving the compounds according to the invention showed a tendency (P = 0.07, T-test) to increase the period t _{s of} active swimming until the first freeze (time A in Fig. 4a), which may indicate their initial slightly lesser depression or greater stamina.

Thus, based on the screening of the compounds according to the invention in tests characterizing the training and psychophysiological state of animals, it was found that these compounds improve long-term memory in a passive avoidance test, significantly reduce animal anxiety and show a tendency to decrease depression. In other words, the compounds of the invention positively affect the learning ability of animals (cognitive function) and the psychophysiological state of animals.

findings

Thus, direct in vivo experiments have shown that cyclic bioisosteres of derivatives of the purine system modulate the nitrergic and dopaminergic systems of the animal brain.

It has been experimentally shown that these compounds can be used as neuroprotectors in pathological conditions of the nervous system. The administration of the compounds of the invention improves cognitive function and psychophysiological state, reducing anxiety and depression.

Compounds according to the invention significantly improve sexual function, and also have a positive effect in withdrawal symptoms.

Given the mechanism of action of the cyclic bioisosteres of the purine system derivatives according to the invention and the involvement of the dopaminergic and nitrergic systems modulated by them in the pathogenesis of various diseases, it can be concluded that the above compounds can be used to treat a number of diseases, including chemical dependencies - disorders caused by substance abuse, such as dependencies from drugs, alcohol and nicotine, sleep disturbances, sexual dysfunctions (including sexual dysfunctions), dying reintestinal disorders, psychoses, affective disorders, inorganic psychoses, personality disorders, psychiatric mood disorders, schizophrenia and schizoaffective disorders, polydipsia, bipolar disorders, dysphoric mania, anxiety and related diseases, obesity, bacterial infections of the central nervous system, such as learning disabilities, memory disorders, Parkinson's disease, depression, extrapyramidal side effects of antipsychotics, hypothalamic-pituitary disorders a, vascular and cardiovascular diseases, dystonia, dyskinesias, hyperkinesia, dementia, ischemia, motor disorders, hypertension and diseases caused by a hyperactive immune system, such as allergies and inflammations, in mammals, including humans.

Claims (13)

1. The use of cyclic bioisosteres of derivatives of the purine system having the general structural formula

where R =

atom Li, Na, K, R ¹ = —H, —NH ₂ , —Br, —Cl, —OH, —COOH, B = -N =, -CH =, Z = -CH =, -N =, A = -N = with B = -N =, Z = -CH-, A = -CH- with B = -N =, Z = -CH-, A = -CH- with B = -N =, Z = -N =, A = -CH- at B = -CH =, Z = -CH =, A = -CH = at B = -CH =, Z = -N =, and their pharmacologically acceptable salts as active ingredients having activity against the nitrergic and dopaminergic systems, in a pharmaceutical composition for treating disorders caused by disorders of the nitrergic system and / or the dopaminergic system of the body, containing the active ingredient in an amount sufficient to affect these systems, in a pharmaceutically acceptable carrier. 2. The use according to claim 1, characterized in that the active ingredient is a pyrido [2,3-d] -6H-pyridazine-5,8-dione derivative having the general formula

where R =

atom Li, Na, K, R ¹ = —H, —NH ₂ , —Br, —OH, —COOH. 3. The use according to claim 1, characterized in that the active ingredient is selected from the group including: 7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (1), 4-amino-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (2), 3-bromo-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (3), 4-hydroxy-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazine-5,8-dione disodium salt (4), 3-carboxy-7- (β-D-ribofuranosyl) pyrido [2,3-d] -6H-pyridazine-5,8-dione disodium salt (5), pyrido [2,3-d] -6H-pyridazin-5,8-dione lithium salt (6), pyrido [2,3-d] -6H-pyridazin-5,8-dione sodium salt (7), pyrido [2,3-d] -6H-pyridazin-5,8-dione potassium salt (8). 4. The use according to claim 1, characterized in that the active ingredient is a derivative of benzo [d] -3H-pyridazine-1,4-dione, having the general formula:

where R =

atom Li, Na, K, R ¹ = —H, —NH ₂ , —Cl, —OH, —COOH. 5. The use according to claim 1, characterized in that the active ingredient is selected from the group including: 2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (9), 5-amino-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (10), 6-amino-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (11), 5-chloro-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione sodium salt (12), 5-hydroxy-2- (β-D-ribofuranosyl) benzo [d] -3H-pyridazin-1,4-dione disodium salt (13), 5-amino-benzo [d] -3H-pyridazin-1,4-dione lithium salt (14), 5-amino-benzo [d] -3H-pyridazin-1,4-dione sodium salt (15), 6-amino-benzo [d] -3H-pyridazin-1,4-dione potassium salt (16), 5-hydroxy-benzo [d] -3H-pyridazin-1,4-dione disodium salt (17), 6-carboxy-benzo [d] -3H-pyridazine-1,4-dione disodium salt (18). 6. The use according to claim 1, characterized in that the active ingredient is a pyrazino [2,3-d] -6H-pyridazine-5,8-dione derivative having the general formula:

where R =

atom Li, Na, K, R ¹ = —H, —NH ₂ , —Br, —OH, —COOH. 7. The use according to claim 1, characterized in that the active ingredient is selected from the group including: 7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (19), 2-amino-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (20), 3-amino-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (21), 3-bromo-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (22), 2-hydroxy-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazine-5,8-dione disodium salt (23), 2-carboxy-7- (β-D-ribofuranosyl) pyrazino [2,3-d] -6H-pyridazine-5,8-dione disodium salt (24), pyrazino [2,3-d] -6H-pyridazin-5,8-dione lithium salt (25), pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (26), 3-bromo-pyrazino [2,3-d] -6H-pyridazin-5,8-dione potassium salt (27), 2-amino-pyrazino [2,3-d] -6H-pyridazin-5,8-dione sodium salt (28). 8. The use according to claim 1, characterized in that the active ingredient is a pyrimido [4,5-d] -6H-pyridazine-5,8-dione derivative having the general formula:

where R =

atom Li, Na, K, R ¹ = —H, —NH ₂ , —Br, —OH, —COOH. 9. The use according to claim 1, characterized in that the active ingredient is selected from the group including: 7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazine-5,8-dione sodium salt (29), 2-amino-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (30), 4-amino-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (31), 2-bromo-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (32), 4-hydroxy-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (33), 4-carboxy-7- (β-D-ribofuranosyl) pyrimido [4,5-d] -6H-pyridazine-5,8-dione sodium salt (34), pyrimido [4,5-d] -6H-pyridazin-5,8-dione lithium salt (35), 2-amino-pyrimido [4,5-d] -6H-pyridazin-5,8-dione sodium salt (36), 4-bromo-pyrimido [4,5-d] -6H-pyridazin-5,8-dione potassium salt (37). 10. The use according to claim 1, characterized in that the active ingredient is used as a neuroprotective agent in a pharmaceutical composition for protecting the nervous system. 11. The use according to claim 1, characterized in that the active ingredient is used in a pharmaceutical composition to improve cognitive function and normalize the psychophysiological state. 12. The use according to claim 1, characterized in that the active ingredient is used in a pharmaceutical composition with anxiolytic and antidepressant effects. 13. The use according to claim 1, characterized in that the active ingredient is used in a pharmaceutical composition for the treatment of diseases selected from the group including: disorders caused by substance abuse, such as addiction to drugs, alcohol and nicotine; sleep disturbances; sexual disorders, including sexual dysfunctions; gastrointestinal disorders; psychoses; affective disorders; inorganic psychoses; personality disorders; psychiatric mood disorders; schizophrenia and schizoaffective disorder; polydipsia; bipolar disorder; dysphoric mania; anxiety and related diseases; obesity; bacterial infections of the central nervous system - such as meningitis; learning disabilities; memory impairment; Parkinson's disease; neurodegenerate diseases, for example, Alzheimer's disease; Depression extrapyramidal side effects of antipsychotics; hypothalamic-pituitary disorders; vascular and cardiovascular diseases; dystonia; dyskinesia; hyperkinesia; dementia ischemia; motor impairment; hypertension and diseases caused by a hyperactive immune system, such as allergies and inflammations, in mammals, including humans, in an amount effective for treatment.

Images