EA006604B1 - Combination of serotonin agonist (5-ht) and antagonist (5-ht) as pharmaceutical formulation - Google Patents

Abstract

The invention relates to a method of preventing or treating a disease related to the 5-HT2C receptor and the 5-HT6 receptor, comprising administering to a human or animal subject in need thereof a 5-HT2C receptor agonist and a 5-HT6 receptor antagonist in sufficient amounts to provide a therapeutic affect. The invention also relates to a pharmaceutical composition comprising an effective amount of a combination of a 5-HT2C receptor agonist and a 5-HT6 receptor antagonist, and optionally a pharmaceutically acceptable carrier.

Description

FIELD OF THE INVENTION

The present invention relates to the prevention or treatment of a disease associated with receptors

5-HT ₂ s and 5-HT ₆ . The invention further relates to a pharmaceutical composition for therapeutic use comprising a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist.

BACKGROUND OF THE INVENTION

Serotonin (5-hydroxytryptamine or 5-HT) is a key neurotransmitter in the peripheral and central nervous system (PNS and CNS) and is involved in various sensory, motor and behavioral functions, such as regulating nutrition, sleep, body temperature, blood pressure, emotions and cognition . At least 14 different serotonin receptor subtypes formally classified are expressed in the mammalian PNS and CNS; see C1 with yours! a1., no. Vsu .. 1990, 14, 35-37; and Ό. Nousg with! A1., Ryagshaeok Vsu., 1994, 46, 157-203. Serotonergic agonists and antagonists are available for the treatment of a wide range of disorders, including anxiety, depression, hypertension, migraine, obesity, drug abuse and addiction, compulsive disorders, schizophrenia, autism, neurodegenerative disorders (e.g., Alzheimer's disease, parkinsonism and Huntington's chorea) and chemotherapy-induced vomiting.

The subfamily of 5-HT ₂ receptors consists of three subtypes - 5-HT _2A , 5-HT _2B and 5-HT _2C receptors. 5-HT _2C serotonin receptors are expressed in many parts of the brain and are involved in the regulation of food intake (Ooipky S.T., Osk. Vsk., 1995, 3, Zirr. 4, 4493-4623; B1sksgb1ks M.1. S! A1 ., ΟίηЬс1ск. Оск. Мс! АБ., 1999, 1, 207-214). It has been shown that a non-specific 5-HT _2C receptor agonist mchlorophenylpiperazine (t-CPP), which is somewhat preferred for 5-HT _2C receptor, reduces feed intake in mice expressing the normal 5-HT _2C receptor, while the compound loses activity against mice expressing the mutated inactive form of the 5-HT _2C receptor (Tcco !! b.N. c! a1., Upps. 1995, 374, 542-546).

In addition, it is reported that t-CPP and I-22394A azepinoindole, the latter have recently been identified as an agonist of the 5-HT _2C receptor (unpublished data), reduce body weight in people after treatment for two and nine weeks, respectively (\ Wa1k11 A. E.Z .. Rkusyoryagtasodu, 1994, 116, 120122; Zagdsp! R.A. s! A1., Rkusyoryagtasodu, 1997, 133, 309-312; and Sa11ai! Ό.Μ. s! A1., Sigg. Vsk., 1967, 9, 579-581).

Recently, a number of pyrrolo [3,2,1-y] quinoline derivatives have been identified as 5-HT _2C receptor agonists having selectivity for the 5-HT _2A receptor (1kaas M. s! A1., Vuogd. Msb. Syst. Bsy. , 2000, 10, 919-921). It is believed that such compounds represent a new approach to the treatment of obesity and epilepsy.

It is also assumed that the subtype of 5-HT _2C receptors is involved in CNS disorders, such as depression and anxiety (1 sys. G. c! A1., Exors Ors. 1 start !. Aegidk. 1998, 7, 1587-1599; Lsuksp O.S. M. GOgidk, 1999, 2, 109-120). It is also assumed that the subtype of 5-HT _2C receptors is involved in urination disorders such as urinary incontinence (Luxux, O.S.M. Googid, 1999, 2, 109-120).

Also, the 5-HT ₆ receptor (identified in 1993 - Mopcta! A1., Mo1. Ryagtasok, 1993, 43, 320327, and Via! M. c! A1., Vyusst. Vyryuk. Vsk. Sottip., 1993, 193, 269-276) is involved in the regulation of food intake and CNS disorders.

So, for example, in Vp! 1su EU. from! A1., Br. 1. Ryagtasog 1999, 126, 66P, describes a reduction in feed intake in rats by administration of a 5-HT ₆ antagonist. Also, some antidepressants and atypical antipsychotics show a high affinity for the 5-HT ₆ receptor, which suggests the involvement of the 5-HT ₆ receptor in schizophrenia (Vo! S! A1., 1. P11agtaso. Exp. Tyss., 1994, 268, 1403-1410; 31s1dy! S! A1., Eksrs Orsh. Thysg. Ra! Sp! K, 1998, 8, 1217-1224; Voigkop s! A1., Vg. 1. Ryagt., 1998, 125, 15621566 ; Voskk s! A1., Mo1. Ryagtaso 1998, 54, 577-583; 31s1dyy! S! A1., Br. 1. Ryagtaso., 1998, 124, 556-562). In addition, the 5-HT ₆ receptor is associated with conditions of generalized stress and anxiety (Wokyoka c! A1., Ltc Zsk, 1998, 17/18, 1473-1477).

Summary of the invention

According to the present invention, it was unexpectedly found that the combined administration of a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist reduces food intake to a greater extent than the administration of either a single agonist or a single antagonist. Such combined administration of a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist may present therapeutic advantages over treatment with either a single agonist or a single antagonist.

Therefore, in one of its aspects, the present invention relates to a pharmaceutical composition comprising an effective amount of a combination of a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist and, optionally, a pharmaceutically acceptable carrier.

In another aspect, the invention relates to a method for preventing or treating a disease, in particular obesity, associated with a 5-HT _2C receptor and a 5-HT ₆ receptor, comprising administering to a human or animal in need thereof a 5-HT _2C receptor agonist and a receptor antagonist 5-HT ₆ (simultaneously or sequentially) in quantities sufficient to obtain a therapeutic effect.

- 1 006604

In another aspect, the invention relates to the use of a 5-HT ₂ s receptor agonist and a 5-HT ₆ receptor antagonist for the manufacture of a medicament for treating a disease associated with a 5HT _2C receptor and a 5-HT ₆ receptor.

In another aspect, the invention relates to a method for producing a pharmaceutical composition, wherein a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist, combined in a therapeutic amount, are thoroughly mixed with a pharmaceutically acceptable carrier.

In yet another aspect, the invention relates to a product comprising a 5-HT _2C receptor agonist and a 5-HT6 receptor antagonist, as a combined preparation for simultaneous, separate or sequential use in the treatment of a disease, in particular obesity, associated with the 5-HT2C receptor and 5-HT6 receptor.

Brief Description of the Drawings

FIG. 1 shows the effect on the feed intake of mice b / bb after combined administration of a 5-HT _2C receptor agonist (ΡΝυ-183933Ρ; 50 mg / kg, po (oral)) and a 5HT ₆ receptor antagonist (Ρυ-186053Α; 50 mg / kg, 8c (subcutaneously)), as well as the action of one agonist and one antagonist.

FIG. 2 shows the effect on food intake by mice of b / bb after combined administration of a 5-HT _2C receptor agonist (BUT.2938P; 5 mg / kg, §c) and a 5-HT ₆ receptor antagonist (BUT.2918C; 3 mg / kg, 5s ) as well as the action of one agonist and one antagonist.

DETAILED DESCRIPTION OF THE INVENTION

As indicated above, the present invention is based on the unexpected discovery that the combined administration of a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist reduces food intake to a greater extent than the administration of either a single agonist or a single antagonist. Such combined administration of a 5-HT _2C receptor agonist and a 5-HT ₆ receptor antagonist may also present some advantages, for example, in the treatment of obesity compared to treatment with either a single agonist or a single antagonist.

First, combined administration requires lower doses of each compound to produce a similar or improved reduction in food intake than treatment with a single compound.

Secondly, the lower doses required for combined administration can reduce the risk of adverse events.

Thirdly, the lower doses required for combined administration can reduce the risk of tolerance and a tendency to abuse.

Fourth, treatment based on two goals can increase individual therapeutic efficacy relative to treatment based on one goal. The risk of non-response (non-responders) may also be reduced.

The beneficial effects of the combined administration of this invention are useful not only for modulating eating behavior and for treating overweight and obesity, but may also be useful for treating central nervous system disorders such as depression, mania, schizophrenic disorders, anxiety, memory disorders (such as Alzheimer's disease ), migraine, addiction to excessive drug use, convulsive syndrome, personality disorders, post-traumatic stress syndrome and sleep disorders, as well as for the treatment of urinary retention (or even a bladder in general with increased activity), sexual dysfunctions, gastrointestinal upsets and glaucoma.

As used herein, the term 5-HT _2C receptor agonist refers to a compound that causes activation of the 5-HT _2C serotonin receptor. The 5-HT _2C receptor agonist preferably has an affinity constant K _{1 of} less than 50 nM, preferably less than 20 nM, and an intrinsic activity ίη νίίτο, measured as the content of intracellular Ca ²⁺ , is higher than 20%, preferably higher than 50%, relative to 5-HT (1 μm).

Used in this description, the term 5-HT ₆ receptor antagonist refers to a compound that causes a blockade of reactions mediated by the 5-HT ₆ serotonin receptor. The 5-HT ₆ receptor antagonist preferably has an affinity constant K _{1 of} less than 50 nM, preferably less than 20 nM, and an intrinsic activity ίη νίίτο, measured as the content of intracellular cAMP, less than 50%, preferably less than 20%, relative to 5-HT (1 μM) .

Assays of ίη νοτο that can be used to determine the affinity and internal activity, respectively, of 5-HT _2C receptor agonists and 5-HT ₆ receptor antagonists are known in the art and are given below in the experimental part as tests for determining the affinity for 5HT2A and 5- NT ₂ century

As a rule, 5-HT _2C receptor agonists and 5-HT ₆ receptor antagonists should be sufficiently selective so as not to have any significant harmful side effects. However, the terms selective and essential in this context should be interpreted broadly, and their meanings are fairly obvious to those skilled in the art.

The 5-HT _2C receptor agonist preferably has a selectivity for the 5-HT _2C receptor of at least 5, preferably at least 10, more preferably at least 20, relative

- 2 006604 Tel'nykh 5HT _2A receptors, 5HT _2C and 5HT ₆ respectively (measured as the ratio of affinities 5HT _2A / _2C, 5-HT, 5-HT _2B / 5HT _2C and 5HT _6/5 -NT _2C ).

The 5-HT ₆ receptor antagonist preferably has a selectivity for the 5-HT ₆ receptor of at least 5, preferably at least 10, more preferably at least 20, relative to the 5-HT _2A , 5-HT _2B and 5-HT _2C receptors, respectively (measured as the ratio of affinities 5НТ2А / 5-НТ6, 5-НТ2В / 5-НТ6 and 5-НТ2С / 5-НТ6).

Appropriate tests to determine whether a compound is a selective 5-HT _2C receptor agonist or a 5-HT ₆ selective receptor antagonist are known in the art, and as described above, are described below in the experimental part.

Currently preferred 5-HT _2C receptor agonists are classes of compounds of arylpiperazines and piperazinylpyrazines, in particular, compounds described in XVO 00/76984 and Swedish patent applications No. 0004244-0 and 0004245-7, filed November 20, 2000

In particular, for the purposes of the present invention, fumarate (2K.) - methyl-1- [3- [2- (3-pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2-methylpiperazine is used as a 5-HT _2C receptor agonist (VUT.2938P).

Currently preferred 5-HT ₆ receptor antagonists are compounds of the azepinindole class, such as the class of arylsulfonated hexahydroazepinindoles described in νθ 01/05793. Other preferred 5-HT ₆ receptor antagonists are compounds of the arylsulfonylindole class, such as the class of compounds described in Swedish Patent Application No. 0003810-9.

In particular, for the purposes of the present invention, 6-methyl-9- (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride is used as a 5-HT ₆ receptor antagonist -186053A).

5-HT _2C receptor agonists and 5-HT ₆ receptor antagonists can be compounds as such or, where appropriate, their pharmaceutically acceptable salts (acid and base addition salts) or their stereochemical isomeric forms (including optical isomers such as enantiomers and racemates).

The pharmaceutically acceptable addition salts described above are therapeutically active non-toxic acid and base addition salts which the compounds are capable of forming. Compounds with base properties can be converted into their pharmaceutically acceptable acid addition salts by treating the base forms with an appropriate acid. Examples of acids are inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and organic acids such as acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, glycolic acid, maleic acid, malonic acid, oxalic acid, benzenesulfonic acid, toluenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, fumaric acid, succinic acid that, malic acid, tartaric acid, citric acid, salicylic acid, p-aminosalicylic acid, pamoic acid, benzoic acid, ascorbic acid and the like. Examples of base addition salt forms are sodium, potassium, calcium salts and salts formed with pharmaceutically acceptable amines, such as, for example, ammonia, alkylamines, benzathine, and amino acids, such as, for example, arginine and lysine. As used herein, the term addition salt also includes solvates that can form compounds and their salts, such as, for example, hydrates, alcoholates, and the like.

5-HT _2C receptor agonists and 5-HT ₆ receptor antagonists can be prodrugs or forms that can release the desired active ingredient after metabolic transformation ίη νινο. The usual procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in Emdp ίί Rgobgidk, eb. N. Vipbdaagb, ΕΙδονίοΓ. 1985.

5-HT2C receptor agonists and 5-HT6 receptor antagonists for administration purposes can be included in various pharmaceutical forms or in the same dosage form, such as the same tablet, or in separate dosage forms. However, in the latter case, it may be appropriate to place the standard dosage form of the 5-HT2C receptor agonist and the standard dosage form of the 5-HT ₆ receptor antagonist in the same package, for example, in cells of the same blister.

5-HT _2C receptor agonists and 5-HT ₆ receptor antagonists in the form of free bases or salts can be incorporated into suitable forms of galenic preparations, such as compositions for oral administration, for injection, for administration in the form of a nasal spray, or similar forms, in accordance with accepted pharmaceutical procedures. Such pharmaceutical compositions of the invention comprise an effective amount of a 5-HT2C receptor agonist and a 5HT6 receptor antagonist in combination with compatible pharmaceutically acceptable carriers or diluents, as is well known in the art. Carriers can be any inert substance, organic or inorganic, suitable for oral, enteral, rectal, transdermal, subcutaneous or parenteral administration, such as water, gelatin, gum arabic, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium phosphate magnesium, talc, colloidal silicon dioxide, and the like. Such compositions may also contain other

- 3 006604 pharmacologically active substances and conventional additives, such as stabilizers, wetting agents, emulsifiers, flavoring agents, buffering agents, etc.

The compositions of the invention can be prepared, for example, in solid or liquid form for oral administration, such as tablets, pills, capsules, powders, syrups, elixirs, dispersible granules, cachets, suppositories and the like, in the form of sterile solutions, suspensions or emulsions for parenteral administration, sprays, for example, nasal spray, transdermal preparations, for example plasters, etc.

The dose level of each specific 5-HT ₂ s receptor agonist and 5-HT ₆ receptor antagonist and the frequency of doses of a particular combination will vary depending on various factors, including the capabilities of each particular compound used, metabolic stability and duration of action of such a compound, patient age, body weight, general health, gender, nutrition, method and time of administration, excretion rate, combination of drugs, severity of the condition being treated. Daily dosage of 5-HT _2C receptor agonist and receptor antagonist

5-HT ₆ each may range from about 0.001 to about 150 mg / kg body weight, preferably from about 0.01 to about 100 mg / kg body weight, in particular from about 0.1 to about 50 mg / kg body weight, administered once or in multiple doses, for example, doses from 0.01 mg to about 1 g each. Typically, such a combined dosage is given orally, but you can also choose another route of administration, for example, parenteral or rectal. An example of a combined dosage form in the form of tablets may be either form (A) in the form of two separate tablets, i.e. one tablet containing 10, 20 or 50 mg of a 5-HT2c receptor agonist, and one tablet containing 10, 20 or 50 mg of a 5-HT ₆ receptor antagonist, or (B) in the form of a combination tablet containing 10, 20 or 50 mg 5-HT _2C receptor agonist and 10, 20 or 50 mg of 5-HT ₆ receptor antagonist.

The invention is illustrated using the following experimental section, which is not restrictive.

Experimental Section

A. Preparation of Test Compounds

The free base of the 5-HT _2C receptor agonist of the (2K) -methyl-1- {3- [2- (3pyridinyloxy) ethoxy] -2-pyrazinyl} piperazine (ΡΝϋ-183933Γ) fumarate receptor agonist is prepared as described in \\ 'O 00 / 76984. The free base is converted into its salt fumarate, so pl. 126-129 ° C. MS t / ζ 315 (M) ⁺ . Elemental analysis: (С ₁₆ Н ₂ ^ ₅ О ₂ .С ₄ Н ₄ О ₄ ), С, Н, Ν.

The 5-HT ₆ receptor antagonist 6-methyl-9- (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡΝϋ-186053Ά) is prepared as described in \ \ 'About 01/05793.

5-HT _2C receptor agonist fumarate (2K) -1- (3- {2 - [(2-ethoxy-3-pyridinyl) oxy] ethoxy} -2pyrazinyl) -2-methylpiperazine (BUT.2938G) is prepared as described in \\ O 00/76984.

The 5-HT6 receptor antagonist 1- (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT. 5182C) is prepared as described in Swedish Patent Application No. 0003810-9, filed October 20, 2000. Briefly , BUT.5182C is prepared according to the general procedure shown in Scheme 1 below, starting from the commercially available 4-piperazinoindole (compound 1), which goes through steps (a) (c) to form 1- (phenylsulfonyl) -4- (1- piperazinyl) -1H-indole (80% yield). HPLC purity>95%; ¹ H NMR (DMSO-de) δ 9.64 (brs, 2H), 8.00-7.85 (m, 3H), 7.79 (d, 1 = 3.77 Hz, 1H), 7.707.65 (m, 1H), 7.63-7.60 (m, 3H), 7.27-7.22 (m, 1H), 6.95 (d, 1 = 3.76 Hz, 1H), 6, 81-6.77 (m, 1H); 3.30-3.20 (m, 4H); ¹³ C-NMR (DMSO-66) δ 144.79, 137.02, 135.22, 134.62, 129.82, 126.85, 125.63, 125.54, 123.49, 111.15, 107.87, 107.76, 47.81, 42.86; MS (pos. E8-YA) t / ζ 342 (M + H).

Scheme 1

Stage (a): Protection of piperazine by the BOC group according to N4

4-piperazinoindole (1 eq.), ΌΜΑΡ (0.1 eq.) And Εΐ ₃ Ν (4 eq.) Are dissolved in DMF. (BOC) ₂ O (1.1 equiv.) Was added and the reaction mixture was stirred at room temperature (12 hours). DMF was evaporated and the residue was purified by silica gel chromatography using a mixture of chloroform, methanol and ammonia as eluent. HPLC: 100% purity. MS t / ζ 302.2 (M + H).

Step (b): Preparation of Intermediate 3

Intermediate 2 (1.0 eq.) Was dissolved in DMF,% 1 (1.3 eq.) Was added and the suspension was stirred for 0.5 h under nitrogen. Benzenesulfonyl chloride (1.2 equiv.) Was added and the reaction mixture was stirred overnight at room temperature. Volatiles

- 4 006604 The residue was dissolved in ES'M, the solution was washed with saturated No. HCO ₃ solution, dried (Md ₈ O ₄ ), filtered and concentrated to give an oily residue, which was purified by silica gel chromatography using hexane and ethyl acetate (7: 3) as eluent , and tert-butyl-4 [1- (benzenesulfonyl) -1H-indol-4-yl)] - 1-piperazinecarboxylate (3) is obtained. HPLC: 100% purity. Analysis by NMR ( ¹ H and ¹³ C) and MS confirms the established structure.

Stage (s): Removal of the protective BOC group

The BOC group from intermediate compound 3 is removed by dissolving the compound in methanol and then adding ether saturated with gaseous HCl. The hydrochloride (4) is filtered off and dried.

B. Preparation of the pharmaceutical composition

Tablet

Ingredients Mg / tablet one. 5-HT ₂ receptor agonist 10.0 2. 5-HT _b receptor antagonist 10.0 3. Microcrystalline cellulose 57.0 4. Secondary Calcium Phosphate 15.0 5. Sodium starch glycolate 5,0 6. Colloidal silicon dioxide 0.25 7. Magnesium stearate 0.75

The active ingredients 1 and 2 are mixed with ingredients 3, 4, 5 and 6 for about 10 minutes. Then magnesium stearate (7) is added, the resulting mixture is stirred for about 5 minutes and pressed in the form of tablets with or without coating.

C. Receptor Affinity and Efficacy Analysis

5-HT _2C Receptor Affinity Assay

The affinity for the 5-HT _2C receptor is determined in comparative experiments, where the ability of the compound to replace ³ H-labeled 5-HT linked to membranes obtained from transfected HEK293 cell line stably expressing is monitored by serial dilution using technology of scintillation proximity analysis (8PA) human 5-HT _2C receptor protein. Nonspecific binding is determined using 5 μM mianserin.

5-HT2A Receptor Affinity Assay

5-HT _2A receptor affinity is determined in comparative experiments, where the ability of a compound to be serially diluted to replace ³ H-labeled ketanserin or lysergic acid diethylamide (BZE) bound to membranes obtained from transfected CHO cell line stably expressing 5-HT receptor protein _{2A of a} person is monitored by measuring with a scintillation counter the radioactivity of membrane homogenates filtered on glass fiber filters. Nonspecific binding is determined using 5 μM mianserin.

5-HT _2B Receptor Affinity Assay

Affinity for the 5-HT _2B receptor is determined in comparative experiments, where the ability of the compound in serial dilution to replace ³ H-labeled 5-HT linked to membranes obtained from transfected CHO cell line stably expressing human 5-HT _2B receptor protein is monitored using scintillation proximity analysis technology (8PA). Nonspecific binding is determined using 5 μM mianserin.

Efficacy assay for 5-HT _2C receptor

The effectiveness of the agonist with respect to the 5-HT _2C receptor is determined by the ability of the compound to mobilize intracellular calcium in transfected HEK293 cells, which stably expresses the human 5-HT _2C receptor protein, using the GBiO-3 fluorescent dye calcium-forming complex (81dta, St. Louis, Missouri, USA). The relative effectiveness (%) is measured relative to the effectiveness of serotonin at a content of 1 μm.

5-HT ₆ Receptor Affinity Assay

In the radioligand binding assay, [ ³ H] -lysergic acid diethylamide (BZE) is used. The analysis is carried out in 96-well plates by adding 11 μl of the test compound solution at the appropriate dilution (11 serial concentrations of samples are used in duplicate in the analysis), 11 μl of the radioligand solution and 178 μl of the washed mixture of 8PA sensitized MSA granules and membranes in buffer for binding obtained from HEK293 cells containing the cloned human 5-HT ₆ receptor. The plates are shaken for about 5 minutes and then incubated at room temperature for 1 hour. The plates are then loaded into counting cassettes and the pulses are counted in a scintillation counter. The resulting cfp (number of pulses per minute) of specific binding is inserted into a one-site binding model using Cpar

- 5 006604

Slave Ρπδΐη, version 2.0. The established values of 1C _{50 are} converted to the K ₁ values of the affinity constant using the Cheng-Prussoff equation (Syepd U.S. s1 a1., Vusyet. RNagtaso 1973, 22, 3099-3108).

5-HT ₆ receptor efficacy assay

The antagonist capabilities for the 5-HT ₆ receptor are determined by the ability of the compound to antagonize the increase in cAMP caused by 5-HT in HEK293 cells stably expressing human 5-HT ₆ receptor protein using the direct cAMP 8PA screening analysis system (KPA559, Integrate Cytogast Vu1es11, Uppsala, Sweden).

Ό. Feed test

Test compounds

5-HT _2C receptor agonists fumarate (2K) -methyl-1- {3- [2- (3-pyridynyloxy) ethoxy] -2pyrazinyl} piperazine (ΡΝυ-183933Ε) and fumarate (2K) -1- (3- {2 - [(2-ethoxy-3-pyridinyl) oxy] ethoxy} -2pyrazinyl) -2-methylpiperazine (BUT.2938E) is dissolved in physiological saline (0.9% No. 101) and diluted in the same vehicle to an appropriate concentration.

5-HT ₆ receptor antagonists 6-methyl-9- (phenylsulfonyl) -1,2,3,4,5,6hexahydroazepino [4,5-b] indole hydrochloride (ΡNυ-186053A) and 1- (phenylsulfonyl) -4 hydrochloride - (1 piperazinyl) -1H-indole (BUT.5182C) is dissolved and diluted in 25% cyclodextrin.

Fresh solutions are obtained on the day of processing.

Animals

Use male mice aged 8-9 weeks (S57VE / 61Vot ^EEP-ob (ob / ob) Bomholtsgard, Denmark) with the average body weight 45 g. Animals are contained in one cell at 23 ± 1 ° C, humidity of 4060%, and have easy access to water and standard laboratory feed. The light cycle of 12 hours of light / 12 hours of darkness is set by turning off the light at 5 hours of the day. Animals are incubated for at least one week before the start of the study. During the periods of the experiment, animals receive special food (WuZegu, Franktown, New Jersey, USA, dust-free granules with an exact weight of 20 mg each).

Experimental Section

At the beginning of the study, the animals are transferred to special working test cells (the modular system for testing animal behavior, NaBuy, CoiBoigp ΙηκίΓ., Allentown, PA, USA). Such cells contain a feeder with sensors for measuring feed intake, an optical lycometer for recording water consumption, and an infrared monitor for recording total motor activity. Monitors are attached to a computer that continuously monitors and logs events. The food pellets are weighed in an amount necessary for one complete study, and filled with fresh tap water and the water bottles are weighed. Animals are kept in a new environment for them for three days to establish baseline indicators. Animals are weighed at 3 hours at the beginning and at the end of the study. Compounds are administered between 4.20 and 5.00 hours before dark. Three groups of animals receive (ί) a 5-HT ₆ antagonist in 25% cyclodextrin; (ίί) a 5-HT _2C agonist in physiological saline; and (w) a combination of a 5-HT _2C agonist and a 5-HT ₆ antagonist, respectively. When combined, the 5-HT ₆ antagonist or saline is administered 30 minutes before the administration of the 5-HT _2C agonist or 25% cyclodextrin. The fourth group receives, respectively, the media introduced in the same way. The study ends on the fifth day. Weighing is carried out on a Me111eg-To1eyo PK5002 / pK802 balance with an auxiliary computer.

Evaluation of the results

The group for each dose consists of 12-16 animals. The data are corrected for feed loss based on weighing the lost feed over 22 hours and suggest that such loss is proportional to time. Calculations are performed with data before and after processing. Values are expressed as percent of the base feed intake (mean ± standard deviation) for the difference between feed intake before treatment and after 3 hours (5-8 hours after noon), 6 hours (5-11 hours after noon), 12 hours (5 1 p.m. - 5 p.m.), 21 p.m. (5 p.m. - 2 p.m.).

The results shown in FIG. 1 show that the combined treatment with a 5-HT ₆ receptor antagonist ₆ ΡNυ-186053A (50 mg / kg, sc) and a 5-HT _2C receptor agonist No. 0-0-183933Ε (50 mg / kg, orally) reduces feed intake significantly more than compounds taken one at a time. Accordingly, the results shown in FIG. 2 show that the combined treatment with a 5-HT _2C BUT.2938E receptor agonist (5 mg / kg, subcutaneously) and a 5-HT ₆ BUT 5UT receptor antagonist (BUT.5182C (3 mg / kg, subcutaneous) reduces feed intake after 12 and 21 hours after administration, significantly more than the compounds taken one at a time. Thus, it is obvious that combined treatment with a 5-HT2C receptor agonist and a 5-HT6 receptor antagonist reduces feed intake more effectively than treatment with either a single agonist or a single antagonist.

Claims (34)

1. A pharmaceutical composition comprising an effective amount of a combination of a 5-HT _2C receptor agonist, which is an aryl piperazine, such as piperazinylpyrazine and a receptor antagonist - 6 006604 5-HT ₆ selected from azepinoindoles, such as arylsulfonated hexahydroazepinindoles and arylsulfonylindoles or salts, an enantiomer or prodrug of said agonist and / or antagonist and, optionally, a pharmaceutically acceptable carrier. 2. The pharmaceutical composition according to claim 1, wherein the 5-HT _2C receptor agonist has a selectivity for the 5-HT _2C receptor of at least about 10, preferably at least about 20, compared with the 5-HT _2A receptor, receptor 5 -NT _2B and 5-HT ₆ receptor, respectively. 3. The pharmaceutical composition according to claim 1 or 2, in which the 5-HT6 receptor antagonist has a selectivity for the 5-HT6 receptor of at least about 10, preferably at least about 20, compared with the 5-HT2A receptor, the 5- HT2B and 5-HT2C receptor, respectively. 4. The pharmaceutical composition according to claim 1, wherein the 5-HT _2C receptor agonist is fumarate (2K.) - methyl-1- {3- [2- (3-pyridinyloxy) ethoxy] -2-pyrazinyl} piperazine (ΡΝυ- 183933Ε). 5. The pharmaceutical composition according to claim 1, wherein the 5-HT _2C receptor agonist is fumarate (2K) -1- (3- {2 - [(2-ethoxy-3-pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2-methylpiperazine (BUT.2938E). 6. The pharmaceutical composition according to claim 1, wherein the 5-HT ₆ receptor antagonist is 6-methyl-9- (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡNυ-186053A). 7. The pharmaceutical composition according to claim 1, wherein the 5-HT ₆ receptor antagonist is 1- (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT.5182C). 8. A method of obtaining a pharmaceutical composition according to any one of claims 1 to 7, in which a therapeutic amount is a combination of a 5-HT2C receptor agonist, which is an aryl piperazine, such as piperazinylpyrazine and a 5-HT6 receptor antagonist selected from azepinoindoles, such as arylsulfonyl substituted hexahydroazepinindoles and thoroughly mixed with a pharmaceutically acceptable carrier. 9. The method of claim 8, wherein the 5-HT _2C receptor agonist is (2K) methyl-1- {3- [2- (3pyridinyloxy) ethoxy] -2-pyrazinyl} piperazine fumarate (ΡΝυ-183933Ε). 10. The method of claim 8, wherein the 5-HT2C receptor agonist is (2K) -1- (3- {2 - [(2ethoxy-3-pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2-methylpiperazine fumarate (VUT.2938E). 11. The method of claim 8, wherein the 5-HT ₆ receptor antagonist is 6-methyl-9 (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡNυ -186053A). 12. The method of claim 8, wherein the 5-HT ₆ receptor antagonist is 1 (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT.5182C). 13. A product containing a 5-HT _2C receptor agonist, which is an arylpiperazine, such as piperazinylpyrazine and a 5-HT6 receptor antagonist, selected from azepinoindoles, such as arylsulfonated hexahydroazepinindoles and arylsulfonylindoles as a combined treatment for the simultaneous use of the drug, associated with the 5-HT _2C receptor and the 5-HT ₆ receptor. 14. The product of claim 13, wherein the 5-HT _2C receptor agonist is (2K) methyl-1- {3- [2 (3-pyridinyloxy) ethoxy] -2-pyrazinyl} piperazine fumarate (ΡΝυ-183933Ε). 15. The product of claim 13, wherein the 5-HT _2C receptor agonist is fumarate (2K) -1- (3- {2 - [(2ethoxy-3-pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2- methylpiperazine (BUT.2938E). 16. The product of claim 13, wherein the 5-HT ₆ receptor antagonist is 6-methyl-9 (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡNυ -186053A). 17. The product of claim 13, wherein the 5-HT ₆ receptor antagonist is 1 (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT.5182C). 18. The product according to item 13, where the disease is selected from among nutritional disorders. 19. The product according to p, where the disease is overweight or obesity. 20. The use of a 5-HT _2C receptor agonist, which is an aryl piperazine, such as piperazinylpyrazine and a 5-HT ₆ receptor antagonist selected from azepinindoles, such as arylsulfonated hexahydroazepinindoles and arylsulfonylindoles for the manufacture of a medicament for the treatment of 5-T ₂ receptor-associated and 5-HT ₆ receptor. 21. The use according to claim 20, wherein the 5-HT _2C receptor agonist is (2K) methyl-1- {3- [2- (3pyridinyloxy) ethoxy] -2-pyrazinyl} piperazine fumarate (ΡΝυ-183933Ε). 22. The use according to claim 20, where the agonist of the 5-HT _2C receptor is fumarate (2K) -1- (3- {2 - [(2-ethoxy3-pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2-methylpiperazine (VUT.2938E). 23. The use according to claim 20, wherein the 5-HT ₆ receptor antagonist is 6-methyl-9 (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡNυ -186053A). 24. The use of claim 20, wherein the 5-HT ₆ receptor antagonist is 1 (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT.5182C). 25. The use according to any one of claims 20-24, wherein the disease is selected from among nutritional disorders. 26. The use according to any one of paragraphs.20-25, where the disease is overweight or obesity. - 7 006604 27. A method of preventing or treating a disease associated with a 5-HT _2C receptor and 5HT ₆ receptor, comprising administering to a human or animal in need thereof a 5HT2C receptor agonist, which is an aryl piperazine, such as piperazinylpyrazine and a 5-HT6 receptor antagonist selected from azepinoindoles, such as arylsulfonated hexahydroazepinindoles and arylsulfonylindoles in sufficient quantities to provide a therapeutic effect. 28. The method according to item 27, where the agonist of the 5-HT _2C receptor is fumarate (2K) -methyl-1- {3- [2- (3pyridynyloxy) ethoxy] -2-pyrazinyl} piperazine (ΡΝυ-183933Ρ). 29. The method of claim 27, wherein the 5-HT _2C receptor agonist is fumarate (2K) -1- (3- {2- [2- (ethoxy-3pyridinyl) oxy] ethoxy} -2-pyrazinyl) -2-methylpiperazine (VUT.2938P). 30. The method according to item 27, wherein the 5-HT6 receptor antagonist is 6-methyl-9 (phenylsulfonyl) -1,2,3,4,5,6-hexahydroazepino [4,5-b] indole hydrochloride (ΡΝυ- 186053Α). 31. The method according to item 27, in which the antagonist of the 5-HT ₆ receptor is 1 (phenylsulfonyl) -4- (1-piperazinyl) -1H-indole hydrochloride (BUT.5182C). 32. The method according to any one of claims 27-31, wherein the disease is selected from among nutritional disorders. 33. The method according to p, where the disease is overweight or obesity. 34. The method according to any one of claims 27-33, wherein the 5-HT _2C receptor agonist and the 5-HT ₆ receptor antagonist are administered as a combined pharmaceutical composition.