UA73331C2 - A method for the treatment of pain and a pharmaceutical composition - Google Patents

Abstract

A method for the treatment of pain is disclosed comprising administration of a pain-ameliorating effective amount of any compound according to structural diagram (I), wherein: A, D and R1 are as defined in the specification. Also disclosed are pharmaceutical compositions comprising a pain-ameliorating effective amount of a compound in accord with structural diagram (I). , I

Description

Description of the invention

The invention relates to the treatment of pain or nociception.

Pain is a sensory sensation, different from the sensation of touch, pressure, heat and cold. It is characterized by those who suffer from it as sharp, dull, prickly, cutting, or burning, and usually includes both the sensation itself and the reaction to it. This range of sensations, as well as the different perception of pain by different individuals, makes it difficult to define pain, however, there are many individuals who suffer from severe and long-lasting pain.

Pain caused by damage to neural structures often manifests itself as hypersensitivity or hyperalgesia and is often called "neuropathic." Pain can also be "caused" by stimulation of nociceptive receptors and transmission through intact neural pathways. Such pain is called nociceptive.

The level of stimulation at which pain becomes noticeable is called the "pain threshold". Analgesics are pharmaceutical agents that relieve pain by raising the pain threshold without loss of consciousness. After the introduction of analgesic drugs, the sensation of pain becomes noticeable only after more intense or longer stimulation. In those who suffer from hyperalgesia, the analgesic may cause antihyperalgesia. Unlike analgesics, such agents as local anesthetics block the transmission in the peripheral nerve fiber and thereby block the sensation of pain. General anesthetics reduce the sensation of pain due to loss of consciousness.

There have been reports that tachykinin antagonists cause antinociception in animals, which is believed to be analogous to analgesia in humans (Madai ei ai., U. Atsiop. Rpaptasoi, 1993, 13, 23-93). In particular, it was shown that non-peptide antagonists of the MK-1 receptor cause such analgesia. For example, the antagonist KR 67580 of the MK-1 receptor caused analgesia at a level close to morphine (Saitei ei aiYi., Rgos. Mai. Asad. Zsi. O5A, 1993, 88,. 10208-10212).

Opioid analgesics are a well-known class of analgesic agents with morphine-like action. Synthetic and semi-synthetic opioid analgesics are derivatives of five chemical classes of compounds: phenanthrenes, phenylheptylamines, he)phenylpiperidines, morphinans and benzomorphans. These compounds have different pharmacological activities: some are strong agonists at opioid receptors (e.g., morphine), others are weak or moderate agonists (e.g., codeine), or show mixed agonist-antagonist activity (e.g., nalbuphine), or are partial agonists ( eg nalorphine). Although such an opioid partial agonist as nalorphine (an M-alkyl analogue of morphine) antagonizes the analgesic effect of morphine, administered alone, it can be a strong analgesic. co

Among opioid analgesics, morphine has the most widespread use, but, along with therapeutic qualities, it has a number of disadvantages, including respiratory depression, decreased gastrointestinal peristalsis, nausea and vomiting. Tolerance and physical dependence also limit the clinical use of opioid compounds. co

Aspirin and other salicylate compounds are often used in therapy to interrupt the increase in inflammation in rheumatic diseases and arthritis and to temporarily reduce pain. Other therapeutic compounds designed for this include phenylpropionic acid derivatives such as ibuprofen and naproxen, sulindac, phenylbutazone, corticosteroids, antimalarials such as chloroquine and hydroxychloroquine sulfate, and phenemates (9). Novzr RIagt., 36:622 (May 1979). These compounds, however, are "less effective against neuropathic pain." With 70 Existing pain therapies also have drawbacks. Some therapeutic agents require prolonged use before the patient feels their effect. Other medicines cause serious side effects in certain patients who

"Iz" must be closely monitored in order to prevent the occurrence of such phenomena in a timely manner. Most existing medications only provide temporary pain relief and require constant daily or weekly use. As the disease progresses, the amount of medication needed to relieve pain often increases, increasing the potential for harmful side effects. and MOMA receptors are determined by binding of M-methyl-O-aspartate and include a receptor/on channel complex with several different binding domains. The MOMA molecule is structurally similar to glutamate (CI), which binds in the glutamate site, and is a highly selective and potent activator of the MOMA receptor (U/Aikipz o (1987), OIpeu (1989)). so 20 It is known that some compounds can bind in the MOMA/ZIsh binding site (for example, SRR, OSSR-en,

СОР 40116, СОР 37849, СО5 19755, MRS 12636, MRS 17742, 0-АР5, O-АР7, СОР 39551, СОР-43487, т MO -100,452, I M-274614, | U-233536 and I 233053). Other compounds are called non-competitive antagonists

MOMA binds in other sites of the MOMA receptor complex (examples are phencyclidine, dizocilpine, ketamine, tilethamine, CM 1102, dextromethorphan, memantine, kynurenic acid, SMOX, OMOX, 6,7-OSOX, 52 6,7-OSNOS, K (--НА-966, 7-chlorokynurenic acid, 5,7-YUSKA, 5-iodo-7-chloro-kynurenic acid, MY -28,469,

HF) МО1И-100,748, МО1-29,951, 1-689,560, 1-687,14, ASRS, ASRSSM, ASRPSE, arcaine, diethylenetriamine, 1,10-diaminodecane, 1,12-diaminododecane, ifen-prodil and 51-82.0715) . These compounds have been reviewed in detail by Kodamuzki (1992) and Mazzei et al. (1993) and in the works cited here.

In addition to its physiological functions, glutamate (IS) can be a neurotoxin. Its neurotoxicity 60 is called "excitotoxicity", since the mediator of both its neurotoxic and therapeutic effects is the process of excitation (OiIpeu (1990), Spoi (1992)).

Usually, when SiI is released at a synaptic receptor, it binds only temporarily and is then quickly removed from the receptor by a process that transports it back into the cell. Under abnormal conditions, for example, stroke, epilepsy, and trauma to the central nervous system, the removal of Si does not occur and it accumulates in the receptor, causing the constant excitation of electrochemical activity that kills the neuron that has Si receptors. In the CNS, numerous neurons have SI receptors and therefore excitotoxicity can cause very significant damage to the CNS.

Acute excitotoxic injury can occur as a result of ischemia, hypoxia, brain or spinal cord injury, certain food poisonings associated with an excitotoxic poison such as domoic acid, seizure-induced neuronal degeneration, which may result from persistent epileptic seizure activity ). Considerable evidence supports that the MOMA receptor is one subtype of receptor through which SI mediates multiple CNS injuries, and that MOMA antagonists are effective in protecting neurons

CNO from excitotoxic degeneration in these acute traumatic syndromes of the central nervous system (Stpoi (1988), OIpeu (1990)).

In addition to the neuronal damage caused by acute strokes, overactivation of SH7/0 receptors may contribute to a more gradual neurodegenerative process that leads to cell death in various chronic neurodegenerative diseases, including Alzheimer's disease, amyotrophic lateral sclerosis, AIDS-related dementia, Parkinson's disease and Huntington's disease (Oipeu (1990)). It is believed that MOMA antagonists may be useful in the treatment of such chronic diseases.

In 1980 PCR (also known as "angel dust") was found to act as a "PCR recognition site" in the 7/5 ion channel of the MOMA Sim receptor. PCR acts as a non-competitive antagonist that blocks the flow of ions through the MOMA ion channel. It was later discovered that drugs that act at the PCR site as noncompetitive MOMA antagonisms can produce psychotomimetic side effects. In addition, it was found that certain competitive and non-competitive MOMA antagonists can cause similar pathomorphological changes in the rat brain (Oipeu ei ai. (1991),

Nagogeamez her a). (1993)). Such compounds also have a psychotomimetic effect on the human brain (Kgiziepvzep ei ai. 20. (1992), Netipo (1994), Sgotsa (1994)).

The glycine binding site of the MOMA receptor complex can be distinguished from the binding sites of sow and PCR. In addition, it was recently discovered that MOMA receptors exist in the form of several subtypes that differ in the features of the glycine binding sites of the receptor. Compounds that bind to the glycine site of the MOMA receptor and can be used to treat stroke and neurodegenerative conditions are described in US Pat. and 6,103,721.

It has been found that certain compounds capable of binding to the glycine site of the MOMA receptor can i) relieve pain, particularly neuropathic pain.

Therefore, the object of the invention is a method of treatment for pain, which consists in the introduction for pain relief of an effective amount of any compound of the structural formula I: "- in I co

M M so what and -

And where А is (СНа)р (n has the value 0, 1, 2, З or 4), О is a 5- or б--substituted heteroaryl group or its benzo derivative, which has 1, 2 or З oxygen atoms in the ring or nitrogen or sulfur, and B is a halogen. "

In specific embodiments, the method according to the invention consists in the introduction of an effective pain-relieving amount of a compound of structural formula I, in which OO is selected from the group consisting of pyridyl, quinolyl, pyrazinyl, t c pyridizinyl, furanyl, benzI|p|furanyl, imidazolyl, oxazolyl, thienyl, benz|(B)hienyl and thiazolyl. h In one of the embodiments, the method according to the invention consists in introducing an effective pain-relieving amount » of the compound of the structural formula II:

AND

-I and: whether 9) he is at 50 Pp. ah Some embodiments of the invention include a method of treatment with compounds of structural formula II wherein O is selected from the group consisting of pyridyl, quinolyl, pyrazinyl, pyridizinyl, furanyl, benz|furanyl, imidazolyl, oxazolyl, thienyl, benz|(b)hyenyl, and thiazolyl

Certain embodiments of the invention include a method of treatment with a typical compound described herein.

Other objects of the invention are pharmaceutical compositions containing a compound of the structural formula I;

GF) use of compounds of the structural formula | for the preparation of medicines and pharmaceutical compositions; and which method of joining the compound according to the invention to the glycine site of the MOMA receptor of a warm-blooded animal, for example, a human, for therapeutic inhibition of the activity of the MOMA receptor.

Compounds according to the invention are those described in the description and, in particular, those marked as typical. 60 Suitable pharmaceutically acceptable salts of the compounds of the invention include such acid addition salts as methanesulfonate, fumarate, hydrochloride, hydrobromide, citrate, tris(hydroxymethyl)aminomethane, maleate and phosphoric acid salts of sulfuric acids. In other embodiments, base salts such as alkali metal salts such as sodium, alkaline earth metal salts such as calcium or magnesium, organic amine salts such as triethylamine, morpholine, M-methylpiperidine, M-ethylpiperidine, procaine, dibenzylamine, choline are suitable. , because M,M-dibenzylethylamine or amino acids, for example, lysine.

Another object of the invention is the method of preparation of compounds according to the invention, which includes operations: a) preparation of Bos-protected hydrazine according to the procedure according to the following scheme: o vosMnmn, shi rod 0 V TNEabomeon, SV t 1095 RI/S, MEeOH

H, 2.8 at, 2-6 p.m. o os In MNMYVvoOs

Y -- Zhi r Kk or Kk

Another option: Ry

Х Basis that Ж c) B

HEE SI, Vgabo OMB5; K - H or alkyl b) addition of the indicated Boc-protected hydrazine and cyclization of the product in accordance with the following scheme with the above to form a compound of structural formula 1:

MiMNVos n o

An "/ ВН о in Vos-M, ра - СМС or another similar оно ш- connecting reagent 8-х - о - - о

M so in! 5 on M s - o 35 . sno «7 rch- ' M TNE, VT s no ra . and: M o. " y - r. - c "» on " where CMK is metho-p-toluenesulfonate of 1-cyclohexyl-3-(2-morpholineethyl)carbodiimide; group K/H/O - grouping -A-O of structural formula I; IN! - as defined for structural formula I.

For use of a compound of the invention or a pharmaceutically acceptable salt thereof for the treatment or prevention of pain in a mammal, including humans, the compound may be incorporated into a pharmaceutical composition according to standard pharmaceutical practice. Pharmaceutical compositions containing the compound of the invention can be administered by conventional routes, for example, orally, locally, parenterally, sublingually, nasally, vaginally

Mom either rectally or by inhalation. For this, the compounds according to the invention can be produced in the form of, for example, tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, jellies, nasal injections, suppositories, fine powders or aerosols for inhalation, and sterile aqueous or 7" oily solutions or sterile suspensions or emulsions for parenteral administration (including intravenous, intramuscular or infusion). The preferred route of administration is by oral tablet or capsule.

In addition to the compound of the invention, the pharmaceutical composition according to the invention may contain one or more others

GF) pharmacologically active agents, or such pharmaceutical compositions can be simultaneously or sequentially administered together with one or more other pharmacologically active agents. o Pharmaceutical compositions according to the invention should be administered to the patient in effective pain-relieving daily doses. The daily dose, if necessary, can be divided into separate doses, and the exact amount of compound 60 when taken and the method of administration depends on the weight, age and sex of the patient receiving treatment, and on the specific stage of the disease, according to the principles of humor. It is preferable to take it once a day.

Another embodiment of the invention includes a pharmaceutical composition comprising a compound of structural formula I as defined above, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable additive, such as an excipient or carrier. because Another embodiment of the invention provides for the use of a compound of the structural formula I, or its pharmaceutically acceptable salt for the preparation of a medication intended for attachment to the glycine site of the receptor

MOMA in warm-blooded animals, for example, humans.

One of the embodiments of the invention includes a method of joining the compound of the invention to the glycine site of the receptor

MOMA in a warm-blooded animal, such as a human, in need of treatment for pain which involves administering to that animal an effective amount of a compound of structural formula I or a pharmaceutically acceptable salt thereof.

Definition

The term "alkyl" here includes straight and branched chain alkyl groups, but single chain groups correspond to individual alkyl groups, for example, "propyl". 70 Halogens include fluorine, chlorine, bromine, and iodine.

The term "allyl" herein includes an unsaturated carbon ring or a benzo derivative thereof. In particular, aryl means phenyl, naphthyl, or biphenyl, most often phenyl.

The term "heteroaryl" or "heteroaryl ring" means, unless otherwise specified, a mono-, bi-, or tricyclic (5-15)-monounsaturated or partially unsaturated ring containing up to 5 heteroatoms selected from 7/5 of nitrogen, oxygen of sulfur, and the -СНо- group can optionally be replaced by -С(0)-, and the nitrogen atom in the ring can optionally be oxidized to form M-oxide. Examples of such heteroaryls can be thienyl, furyl, pyranyl . , indazolyl, benzofuranyl and cinolinyl.

The term "heterocycle" or "heterocyclic ring", unless otherwise specified, means a mono-, bi- or tricyclic (5-15)-one fully saturated ring containing up to 5 heteroatoms selected from nitrogen, oxygen, and sulfur, wherein the group -СНо- can alternatively be replaced by -С(0)-. Examples of such heterocycles are morpholinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, piperazinyl, homopiperidinyl, homopiperazinyl, and quinuclidinyl.

If the optional substituents are selected from "one or more" groups, this includes compounds where all of the substituents i) are selected from one of the defined groups, and compounds where the substituents are selected from one or more of the defined groups.

In general, in the methods, processes and examples presented here: concentration is carried out by centrifugal evaporation of the mass; "- operations are carried out at an external temperature of 18-262C in a nitrogen atmosphere; column chromatography (flash procedure), unless otherwise specified, is performed on silica Megsk i.

Kieseidei (Ag. 9385); so outputs are given for illustration only and do not correspond to the maximum possible; the structure of the final products of the formula | are usually determined using NMR and mass spectroscopy, and. proton magnetic resonance is determined in DMSO-th; with the use of the Magiap Setipi 2000 spectrometer, which works in the ZOOMGhzC field; chemical shifts are given in parts per million below tetramethylsilane as an internal standard (scale 5). Peak multiplets are marked as follows: 5 - singlet, b85 - wide singlet, y - doublet, AB or я - double doublet, Я - triplet, ди - double triplet, т - multiplet, bt " - wide multiplet. Mass spectrographic data during the bombardment with fast atoms (RAV) are obtained by the Riayogt spectrometer from Misgotazz in the electron beam, and data on positive and negative ions were collected, in this case (MAN) is given." Infrared spectrum data are obtained with the help of Misoie (Amayag 360 ET-IC.) Intermediate compounds are usually not fully characterized, and purity is determined by mass spectrography or NMR analysis.

In the following, the following abbreviations and definitions are used: - and SOSI" - deuterated chloroform; syu CMC -1-cyclohexyl-3-(morpholineethyl)carbodiimide-metho-p-toluenesulfonate;

DCM - dichloromethane; (95) DHLM - dichlorurea; syu 50 DCC - 1,3-dicyclohexylcarbodiimide;

DMAP - 4-(dimethylamino)pyridine; - DMF - M,M-dimethylformamide;

DMSO - dimethylsulfoxide; t/v - mass spectrography;

MMP - M-methylpyrrolidone; o NMR - nuclear magnetic resonance;

THF - tetrahydrofuran; The given examples and tests illustrate the invention without limiting it.

Examples 60 Example 1. Methanesulfonate of 7-chloro-4-hydroxy-2-(2-pyridylmethyl)-1,2,5,10-tetrahydropyridazine(i4.5-b)quinoline-1.10-dione (tert-butoxy)-M -(4-pyridylmethyl)amino|carboxamide

Triethylamine (1008 ml, 0.78 mol) and then 4-picolyl chloride hydrochloride (40.0 g, 0.24 mol) were added to a stirred solution of tert-butyl carbazate (175 g, 1.3 bmol) and dry DMF (400 mL) under a nitrogen atmosphere. The reaction mixture 65 is heated for 5 hours. at 759 and allowed to cool to room temperature, then diluted with water (2 L) and extracted with ethyl acetate (4 x 500 ml). The combined ethyl acetate extracts are concentrated under reduced pressure and the residue is dissolved in diethyl ether. The resulting solution is washed with water (Zx 40Oml) and brine (40Oml) and dried over Na»SO, which is then filtered, the filtrate is concentrated under reduced pressure and amber oil (130.4g) is obtained. This product was purified by flash chromatography on silica gel eluting with hexane/ethyl acetate (1:1) to give the final compound as a solid off-white foam (24.46g, 45965). "H NMR (300 MHz, DMSO-a): δ 1.36 (z, 9H), 3.90 (a, 2H, U - 4.0Hz); 5.04 (a, 1H, U - 4.5Hz) 8.48 (y, 1H, y - 4.5 Hz).

Dimethyl-7-chloro-4-hydroxyquinoline-2,3-dicarboxylate

A stirred mixture of methyl-2-amino-4-chlorobenzoate (2.50g, 13.5mmol) and dimethylacetylenedicarboxylate (2.05g, 144mmol) in tert-butanol (22ml) is refluxed for 7 hours. 70 nitrogen in the atmosphere. After addition of dimethyl acetylenedicarboxylate (1.16 g, 8.13 mmol) and holding under reflux for another 2.5 hours. the reaction mixture is left to cool to room temperature and potassium tert-butoxide (1.56 g, 13.9 mmol) is added. After the formation of a precipitate, the mixture is kept under reflux for 1.5 hours, after which it is cooled to room temperature and filtered to separate the solid substance, which is washed with tert-butanol and diethyl ether, dissolved in water and acidified with 1M sulfuric acid 75 to form a precipitate. The mixture was extracted with DCM and the combined extracts were washed with water and brine, dried over NaOH, filtered and concentrated to give a green solid. Recrystallization of this material from methanol gives the final compound (1.15 g, 4795) as a whitish solid, m.p. 2332-2336.

MS (SI): 296 (MAN).

Analysis for C43N.oSIMO 5

By calculation: C, 52.81; H, 3.41; M 4.34;

Obtained: C, 52.75; N, 3.47; M 4.69.

3-carbomethoxy-7-chloro-4-hydroxyquinoline-2-carboxylic acid

An aqueous solution of sodium hydroxide (0.27g, 6.75mmol) was added to a stirred suspension of dimethyl-7-chloro-4-hydroxyquinoline-2,3-dicarboxylate (1.0g, 3.3mmol) in Ce water (20ml). After addition, the suspension dissolves. at

The reaction mixture for an hour. heated to 652C, then cooled to room temperature and acidified with concentrated hydrochloric acid. The product is extracted into diethyl ether and ethyl acetate.

The organic extracts are dried over Na5SO4, filtered and concentrated in vacuo to give a solid final product (900 mg), which is purified by recrystallization from the ethyl acetate/hexane co-solvent system to give the desired compound (571 g, 6095) as a solid, temp. sq. 296 (schedule). co

MS (SI): 238 (M.N).

Analysis for С42НаМО СИ 0.45С2НЗСООСНоСНze0.1Н2О: o

According to the calculation: C, 51.30; H, 3.68; M 4.34; co

Obtained: C, 51.28; H, 3.62; M 3.97;

From TN NMR 8.22 (4, U - 8.7 Hz, 1); 7.92 (9, 1H, U - 1.8Hz); 7.28 (yes, 1H, U - 8.7, 1.8Hz); 3.90 (in, 3H). t

3-carbomethoxy-2-pyrrolidineurea-7-chloro-4-hydroxyquinoline

To a suspension of 3-carbomethoxy-7-chloro-4-hydroxyquinoline-2-carboxylic acid (2.25g, 8.0mmol) in THF (20ml) at room temperature under a nitrogen atmosphere, DCC (1.65g, 8.0mmol) was added and pyrrolidine (0.596 g, 8.4 mmol). "

The reaction is stirred for 15 hours. at room temperature, after which urea is removed by filtration. The product Z7Z 70 is purified by flash chromatography on a column with methanol (b59o) in chloroform as an eluent, and the final product c is obtained (2.52 g, 94.3905) in the form of a light brown solid, temp., m.p. 21590. :z» MS (SI): 335 (MAN). "YAN NMR (300MHz, DMSO-av): 5 8.12 (9, 9 - 8.7Hz, 1H), 7.60 (a, YAN, 9 - 1.8Hz); 7.47 (da, 1H, U - 88, 2.0 Hz); 3.69 (v, ZN), 3.40-3.49 (t, 2H), 3.27-3.33 (t, 2H), 1.80-1, 96 (t, 4H). -I 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)-hydroxyquinoline-3-carboxylic acid

To a suspension of 3-carbomethoxy-2-pyrrolidinecarbamide-7-chloro-4-hydroxyquinoline (2.52 g, 7.5 mmol) in deionized water (40 ml), an aqueous solution (2 ml) of potassium hydroxide (882 mg, 15.75 mmol) was added dropwise. after which the reaction is heated to 6b0eC and through Zhod. filtered to remove insoluble material. The filtrate is acidified to pH-1 and a white precipitate is obtained, which is separated by vacuum filtration, washed with water, and dried at 302 in a vacuum for 1 hour. and obtain the final compound (1.5 g, 6495) in the form of a white solid, - M temp. sq. 225-890,

MSc(c: 321 (MH). "IN NMR (300 MHz, DMSO-av): 5 8.28 (9.94 - 8.8 Hz, 1), 7.77 (in, 1H); 7.64 (9 , 1H, U - 8.7Hz); 3.52-3.57 (t, 2H), 3.17-3.19 (t, 2H), 1.83-1.98 (t, 4H).

Gee! M-C-tert-butoxy)carbonylamino|7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)-(hydroxyquinolyl)|-M-(4-pyridylmethyl)carboxamide de To a stirred mixture of 7-chloro-4-oxo-2-( pyrrolidinylcarbonyl)-hydroxyquinoline-3-carboxylic acid (24.29g, 75.733mmol) and dry THF (1175ml) under a nitrogen atmosphere, add CMK (50.55g, 119.34mmol) in portions (35g each, and 60 after 1 min. for 15 ,55g). After stirring the mixture for 20 minutes. a solution of tert-butoxy-N-(pyridylmethyl)amino|carboxamide (22.0g, 98.5mmol) and THF (580ml) was quickly added and the mixture was stirred overnight, then filtered and the filter cake washed with DCM (ZOOml). The filtrate and washings are combined and additional DCM (800 ml) is added. The resulting solution is washed with water (2 x 50 Oml) and dried over

Ma»5zO,, is filtered under reduced pressure and a yellow foam is obtained. The foam is treated with diethyl ether (800 ml), the mixture is stirred and then filtered. The filter briquette is dried at 4592 in vacuum and the final compound (24.3 g, 6190) is obtained as a yellow powder.

Methanesulfonate of 7-chloro-4-hydroxy-2-(4-pyridylmethyl-1,2,5,10-tetrahydropyridazine-|4,5-B)quinoline-1,10-dione

To the mixed mixture

N-tert-butoxy)carbontamino)|7-chloro-4-oxo-2-(pyrrolidinyl-carbonyl)(hydroxyquinolyl|-M-(4-pyridylmethyl)carboxamide (24.0g, 45.62mmol) and dry THF ( 9bOml) in a nitrogen atmosphere add methanesulfonic acid (12Oml, 177.7g, 1.85mol). The mixture is stirred overnight, filtered and the solid is washed with THF (2x 100ml), methanol (2x 5Oml) and diethyl ether (100ml). Filter briquette ( 13.4g) is suspended in methanol (250ml), the mixture is sonicated and filtered. The collected solid is washed with methanol (2 x 70 LObml) and diethyl ether (10Oml), dried at 4590 in vacuum to give the final compound (12.1g, 5990) in the form of a yellow powder, temperature square 225096,

TN NMR (300 MHz, DMSO-dv): δ 2.32 (8, ZM), 5.36 (in, 2H); 7.49 (ad, IN, U - 81Hz, 2.1Hz), 7.86 (a, 1H, y - 6.6Hz); 8.06 (а, 1Н, У - 2.1 Hz); 8.12 (a, 1H, 9 - 8.1Gu); 8.82 (а, 1Н, У - 6.6Hz); 12.6 (Bg in, 1H), 12.84 (horn 5, 1H).

According to the calculation for С47Н44СИМаОвеSNАЗО»зН»e 0.82: С, 46.47; H, 3.60; M, 12.04. t Obtained: C, 46.39; H, 3.65; M, 11.98.

Example 2. Methanesulfonate of 7-chloro-4-hydroxy-2-(3-pyridylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-b)quinoline-1,10-dione

Tert-butoxy-N-3-pyridylmethyl)amino|carboxamide

Triethylamine (128 ml, 0.92 mol) was added to a solution of tert-butyl carbazate (203.6 g, 1.54 mol) and dry DMF (300 mol) under a nitrogen atmosphere, and then a suspension of 3-picolyl chloride hydrochloride (50.0 g, 0.00 mol) in DMF ( ZOOml).

The reaction mixture is heated according to at 759C, cooled to room temperature and diluted with water (2.4 l), after which it was extracted with diethyl ether (Zh 800 ml). The aqueous layer is saturated with salt and extracted with diethylether (Zh 80Oml). The combined extracts are washed with water (1 L), brine (1 L) and dried over NaCl, which is then filtered off and the filtrate is concentrated under reduced pressure. The product was purified by flash chromatography on silica gel with diethyl ether as eluent to give the final compound (23.3 g, 3495) as a whitish solid (o).

TN NMR (300 MHz, DMSO-iv): 5 1.36 (z, 9M), 3.88 (4, 2H, 9 - 4.0Hz), 7.96 (a, 1H, Y - 4.0Hz); 7.33 (ad, 1n, 9y 7 7Hz, 48Hz); 7.71 (a, 1H, 2 77); 7.44 (a, 1H, U - 4.7Gu); 8.49 (v1N). "-

M-Cteret-butoxy)carbonylamino|| 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroxyquinolyl)-M-(4-pyridylmethyl)carboxamide She

To a stirred mixture of 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)-hydroxyquinoline-3-carboxylic acid of Example 1 (20g, 62.4mmol) and THF (800ml) under a nitrogen atmosphere, CMK (40.0g, 94.4mmol) is added ) and then quickly add a solution of (tert-butoxy)-M-(3-pyridylmethyl)amino|carboxamide (20.9g, 93.bmmol) and THF (450ml) and a mixture of She

35 is stirred overnight, then filtered and the filter briquette is washed with DCM (ZOOml) and the mixture is suspended in DCM and filtered. The filtrates are combined and evaporated under reduced pressure. The residue is dissolved in DMSO, dried over NaCl, filtered under reduced pressure and a foam is obtained. The foam is treated with diethyl ether (200 ml) and filtered. The filter briquette is treated with ultrasound, washed with diethyl ether, dried at 402C in a vacuum, and the final compound (32.8 g, 100905) is obtained in the form of a whitish powder. - with 7-chloro-4-hydroxy-2-(3-pyridylmethyl)-1,2,5,10-tetrahydro-pyridazine(i4,5-b)quinoline-1,10-dione methanesulfonate. -C-tert-butoxy)carbonylamino|7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroxyquinolyl)|-M-(4-pyridylmethyl)carboxamide (32.8 mol) and THF (Tl) under a nitrogen atmosphere are added methanesulfonic acid (150ml, 222Hg, 2.31mol) for 1O0min. The mixture is stirred overnight, filtered and the solid washed with THF. -I The filter briquette (13.4g) is suspended in methanol (25O0ml), the mixture is sonicated (Zohvil.) and The solid substance is resuspended in methanol, treated with ultrasound (ZOxvil.) and filtered, then washed with methanol, dried at 1002 in vacuum and the final compound (19.4 g, 6b9Uo) is obtained in the form of (9) as a white solid, temp. 30020. at 50 TN NMR (300 MHz, DMSO-dv): 5 2.33 (8, ZM), 5.29 (v, 2H); 7.46 (ad, IN, U - 9.0 Hz, 2, 1Hz), 7.94 (a, 1H, y - 95.0Hz, 5.6Hz); 8.04 (a, yin, y - 1.8Hz3); 8.16 (a, yin, y - 8.7Hz ); 8.37 (a, tn, 9 - v1Hz); 8.82 (a, 1nH, U - t 4.8Hz); 8.89 (in, 1H).

According to the calculation for С47Н44СИМаозеСНАЗОзенно: С, 46.11; H, 3.66; M, 11.95.

Obtained: C, 46.34; H, 3.61; M, 11.94. 29 Example C. Methanesulfonate

HF) 7-chloro-4-hydroxy-2-(2-pyridylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-b)quinoline-1,10-dione Tert-butoxy-M-(2 -pyridylmethyl)amino|carboxamide

Triethylamine (128 ml, 0.92 mol) and then 3-picolyl chloride hydrochloride (54.0 g, 0.3 mol) were added to a solution of tert-butyl carbazate (174 g, 0.94 mol) and dry DMF (400 mL) under a nitrogen atmosphere. The reaction mixture for an hour. 60, leave to mix at room temperature, heat Acc. at 702C and allowed to cool to room temperature. Then the mixture is diluted with a mixture of ethyl acetate/diethyl ether (1:11), washed with brine and extracted. The aqueous layer is analyzed by TLC (eluent - 100965 diethyl ether) and extracted several times with ethyl acetate (200 ml) until the product is invisible. The combined organic extracts were washed with brine, dried over NaCl and filtered. The filtrate is concentrated under reduced pressure and amber oil (approx. 100g) is obtained, which is then crystallized. The product was triturated with diethylether/hexanes (1:1), filtered, dried under reduced pressure to give the final compound (33.4g, 4595) as a whitish solid.

TN NMR (300 MHz, DMSO-dv): δ 1.38 (in, 9M); 3.96 (a, 2H, y - 4.0Hz), 4.98 (a, 1H, y - 4.0Hz), 7.24 (ad, 1H, 9 - 7.6vHz, 7.8Hz); 7.48 (a, 1); 7.74 (aa, 1H, y - 7.5Hz, 7.8Hz); 8.32 (in, Bg, 1H), 8.47 (an, 2-4.8ГМ).

M-Ct-butoxy)carbonylamino| 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroxyquinolyl)|-M-(2-pyridylmethyl)carboxamide

To a stirred mixture of 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)hydroxyquinoline-3-carboxylic acid of Example 1 (17.5g, 54./mmol) and dry THF (900ml) under a nitrogen atmosphere, add CMK (35.7g, 81.2 mmol) in portions (25.0 g each, and after 10 minutes - 10.7 g each). After stirring for one hour, a solution of 70. (tert-butoxy)-M-(3-pyridylmethyl)amino|carboxamide (16.5g, 73.9mmol) and THF (400ml) was added to the reaction mixture and the mixture was stirred overnight. The reaction is monitored by TLC (methanol(1095)DCM) and determined to be complete, after which it is filtered and the collected solid is washed with THF. The filtrate and washings are combined and concentrated under reduced pressure. The residue is suspended in an aqueous solution of bicarbonate and brine and extracted with DCM (300 ml).

The extracts are combined with previously concentrated organic extracts and washed with bicarbonate and brine 75 (three times) and dried over Na»3O). Ma»5zO), filtered and the filtrate is concentrated under reduced pressure, obtaining a residue, which is purified by flash chromatography on silica gel (eluent - isopropanol (5Uv)/chloroform). After concentration of the desired fractions in vacuum, the final compound (24.3 g, 6195) is obtained in the form of a light brown powder.

Methanesulfonate of 7-chloro-4-hydroxy-2-(2-pyridylmethyl)-1,2,5,10-tetrahydro-pyridazine(i4,5-b)quinoline-1,10-dione

To the solution

M-Ct-butoxy)carbonylamino| 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroxyquinolyl)|-M-(2-pyridylmethyl)carboxamide (24.0 g, 0.045 mol) and dry THF (800 ml) under a nitrogen atmosphere add methanesulfonic acid ( 10 Oml, 148 g, 1.54 mol) within 1 min. The mixture is stirred overnight, filtered and the solid is washed with THF (2 x 100 ml). The filter briquette (15.8 g) is suspended in methanol (2500 ml), the mixture is treated with "SM ultrasound" (ZOkhvil.) and filtered. The solid substance is washed with methanol (2 x 100 ml) and diethyl ether (100 ml), dried at 352C in a vacuum to obtain the final compound (12.1 g, 5995) in the form of an orange powder, temp. sq. 230020,

TN nMR (300 MHz, DMSO-iv): δ 2.33 (in, 3M), 5.35 (z, 2H); 7.46 (9, 71H, 2 - 8.7Hz), 7.64 (9, IN, U - 7.8Hz); 7.68 (aa, ln, 0 -4.8Gu, 6.6Hz); 8.02 (v, 1H), 8.14 (a, yin, y - 8.7Gguu; 8.19 (ai, Yn, y - 66bgtsu, -- 7.8Hz); 8.73 (y, 1H, U - 4.8 Hz); 10.06 (v, g, 1H), 12.84 (v, Bg, 1H). Go)

According to the calculation for C47H44SIMAOzeSNazOzH: C, 47.95; H, 3.35; M, 12.43. with

Obtained: C, 47.93; H, 3.42; M, 12.01.

Example 4. 7-chloro-4-hydroxy-2-benzo|4|furan-2-ylmethyl-1,2,5,10-tetrahydro-pyridazine|(4,5-BIquinoline-1,10-dione, so

M-(1-aza-2-benzo|a|furan-2-ylvinyl/(tert-butoxy)carboxamide ich-

To a solution of benzofuran-2-carboxaldehyde (5.0g, 34mmol) in THF (200ml) with stirring, add tert-butyl carbazate (4.5g, 34mmol) and then concentrated HCI (10 drops) at room temperature. The reaction is stirred for 24 hours. while removing THF in vacuo, the solid was triturated with hexanes and filtered to obtain the final compound (9g, 10095) as a white solid. " "YAN NMR (ZO0MHz, DMSO-yv): δ 11.12 (rg 8, 1H), 8.02 (v, 1H), 7.67 (9, yin, U - 7.6Hz); 7.62 (a, yin, - Z c 8.5Hz); 7.37 (aa, 1H, y - 7.6Hz, 7.6Hz); 7.27 (aa, tn, U - 7.6Gu, 7.6Hz) ; 7.21 (in, 1H), 1.48 (5, 9H). -2-benzo|4|furan-2-ylvinyl/tert-butoxy)carboxamide (4.0g, 15mmol) in methanol (75ml) were added sodium cyanoborohydride (7.2g, 115mmol) and acetic acid (10ml). The mixture is heated for 4 hours. at 6590.

If the TLC analysis (hexanes/ethyl acetate (1:1)) shows the remains of the starting material, add more cyanoborohydride - | of sodium (approx. 2g). In 2 hours the starting material disappears and the reaction is cooled to room temperature and the methanol is removed in vacuo. The residue is dissolved in ethyl acetate and washed with sat. The mixture was filtered and concentrated to give the final compound (95) (3.4g, 1Zmmol, 8590) as a white solid, which was used without purification in further operations. syu 50 TN NMR (300 MHz, DMSO-iv): δ 8.36 (z, 1M), 7.57 (9, 1H, U - 7.0Hz); 7.51 (a, YAN, U - 7.7 Hz); 7.22 (t, 2H), 6.74 (v, 1H), 5.01 (Bg 5, 1H); 4.00 (in, 2H), 1.37 (in, 9H). "6 M-(M-(benzo|a|furan-2-ylmethyl)|(7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl|carbonylaminoH-tert-butoxy)carboxamide

1-Cyclohexyl-3-(2-morpholineethyl)carbodiimide- meth-p-toluene sulfonate (10.8 g,

Gee! 26bmmol). A solution is added to this bright yellow mixture

M-Kbenzo|a|Ifuran-2-ylmethyl)amino)tert-butoxy)carboxamide (3.3g, 1mmol) and M,M-dimethylaminopyridine de (230mg, 1.9mmol) in THF (25ml) with stirring. The mixture is kept under reflux in a nitrogen atmosphere for 4 hours, then cooled and filtered. The filtrate is concentrated to give a yellow solid foam, which is chromatographed on silica gel (methanol(10950)/CHClCl) to give the final compound as a pale yellow solid, which is used further without characterization. 7-chloro-4-hydroxy-2-benzo|a|furan-2-ylmethyl-1,2,5,10-tetrahydro-pyridazine(i4,5-B)quinoline-1,10-dione

To the mixture

M-(M-(benzo|a|furan-2-ylmethyl)||7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolylcarbonylaminoHt-butyl 65 c)carboxamide (6.2g, 11mmol) in THF (150ml) at room temperature is added a solution of methanesulfonic acid (29ml, 44mmol) in THF (7Oml).The mixture is stirred overnight and then water (approx. 50Oml) is added,

to cause precipitation of the product. The cream-colored solid is collected and washed with water and diethyl ether, after which it is dried at 302 and 50 Ωm of RT. Art. overnight and obtain the final substance in the form of a whitish solid. "IN NMR (300 MHz, DMSO-iv): 5 12.74 (Bg 5, 1H), 11.96 (Bg in, 1H); 8.15 (4, IN, U - 8.8 GHz), 8.04 (a, 1H, U - 1.6Hz); 7.59 (a, MN, y - 7.7Hz); 7.53 (a, YAN, y - 81Hz), 7.44 (aa, yin, 2 - 2.0 Hz, 8.9 Hz); 7.25 (t, 2H), 6.84 (85, 1H); 5.27 (v, 2H).

According to the calculation for SooN.l2MaSiv 0O.1NoSi 0.3СНзЗОЗН: С, 57.45; H, 3.18; M, 9.90.

Obtained: C, 57.61; H, 3.20; M, 9.91.

Example 5. 7-chloro-4-hydroxy-2-(quinolin-4-ylmethyl)-1,2,5,10-tetrahydro-pyridazine(i4,5-B)quinoline-1,10-dione

This compound is synthesized as in Example 4, using quinoline-4-carboxaldehyde as a starting material. "YAN NMR (300 MHz, DMSO-av): δ 12.87 (rg 8, 1 H), 12.10 (rg c, 1H); 9.16 (a, 1H, 9 - 54 Hz), 8.61 ( 4, 1H,

In the same 84 Hz); 8.30 (a, yin, 0 - 8.4 Hz); 8.19-8.15 (t, 2H), 8.08 (a, tn, 0 - 1.8gu, 8.01 (aa, tn, y - 7.5Hz, 7.8Hz); 7.74 ( a, 2H, ОЙ - 554Gu), 7.47 (aa, 1H, y - 1.8Hz, 8.7Hz); 5.84 (5.1H). 12 Example b. 7-chloro-4-hydroxy methanesulfonate -2-(pyrazin-4-ylmethyl)-1,2,5,10-tetrahydropyridazine(I4,5-c)quinoline-1,10-dione 2-chloromethylpiperazine 2-methylpyrazine (1.0 ml, 22 mmol) in carbon tetrachloride (8 Oml) is treated with M-chlorosuccinimide (4.27 g, 31.5 mmol) and benzoyl peroxide (0.26 g, 1.1 mmol). The mixture is refluxed for 7 hours and then cooled to room temperature. The solid is filtered through diatomaceous earth and washed

DXM. The filtrate is washed with saturated aqueous sodium thiosulfate, saturated aqueous sodium bicarbonate and saturated aqueous sodium chloride. The organic layer is dried over Ma»5O, filtered, concentrated under reduced pressure and used in the next reaction. "NN NMR (300 MHz, DMSO-av): 5 4.88 (z, 2M), 12.10 (rg 5, 1H); 9.16 (a, 1H, y - 54Hz), 8.65-8, 68 (t, 2H); sc 75 vvB (v, 1n). (o) (Tert-butoxy)-M-Ipyrazin-2-ylmethyl)amino|carboxamide

To a solution of 2-chloromethylpyrazine (1.2g, 9Uzmmol) and dry DMF (1bml) in methanol (/5ml) was added tert-butyl carbazate (6.3g, 4v8mmol) and triethylamine (2.bml, 19mmol) under a nitrogen atmosphere. The mixture is heated for 17 hours. "- at 752C and cooled to room temperature, diluted with water and extracted with diethylether (5x). The combined extracts are washed with brine and dried over NaCl5O, which is then filtered. The filtrate was concentrated under reduced pressure to give a brown oil (1g), which was flash chromatographed (silica gel, 2-595% methanol gradient in DCM) to give the final compound (1.64g, 7990) as a brown waxy solid. (ze) "H NMR (Z00MhuC, DMSO-Al5, TEA shaking): δ 1.44 (v, 9H), 4.53 (z, 2H); 8.71-8.78 (t, 2H); 8 ,81 (in, 1H). part

M-Ct-butoxy)carbonylamino| 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroxyquinolyl)|-M-(pyrazin-2-yl ethyl)carboxamide

CMK (4.56 g, 10 .8 mmol). After stirring for 20 minutes, the reaction mixture was treated with a solution of s (tert-butoxy)-N-(3-pyrazin-2-ylmethyl)amino|carboxamide (1.6 g, 7 mmol) and DMAP (4 mg, 0.4 ml) in THF (1 Oml). y The mixture is stirred under reflux for 2 days and then cooled to room "" temperature, after which it is filtered and the filter briquette is washed with THF. The combined filtrates and washings are concentrated and a brown foam is obtained, which is chromatographed (silica gel, methanol(290)/DCM) and obtained

The final compound (Z3.1g, 8290). - and Methanesulfonate 7-chloro-4-hydroxy-2-(pyrazin-2-ylmethyl)-1,2,5,10-tetrahydro-pyridazine (4,5-Schiquinoline-1,10-dione

To a stirred mixture of M-tert-butoxy)carbonylamino|7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl)|-M-(4-pyrazin-2-ylmethyl)

Ge) of carboxamide (3.1 g, 5.9 mmol) and dry THF (100 ml) in a nitrogen atmosphere, methanesulfonic acid (13.5 ml, 0.21 mol) is added. The mixture is stirred overnight, filtered and the solid is washed with TIF, after which it is treated with methanol and ultrasound (1 hour). The solid substance is collected by filtration, washed with - M methanol, dried at 502 in vacuum and the final compound (2.0 g, 8095) is obtained as a white powder, temp. sq. 235-245960. "YAN NMR (Z00MHz, DMSO-av): 5 2.38 (RNUSO", 8, ZN), 5.27 (v, 2H); 7.44 (a9, yin, y - 1.8Hz, 8.7Hz ), 8.04 22 (y, 1H, y - 1.8Hu); 8.14 (a, 1H, U - 8.7Hu); 8.58 (da, 1H, y - 2.4Hz, 7.5Hu ); 8.63 (in, 1H).

Ge) According to the calculation for C416NiaoSIM5OzeSNazO»zNe No: C, 43.46; H, 3.43; M, 14.90.

Obtained: C, 43.28; H, 3.34; M, 15,17. o Example 7. 7-chloro-4-hydroxy-2-(5-isoxaline)umethyl-1,2,5,10-tetrahydro-pyridazine(i4,5-b)quinoline-1,10-dione

Tert-butyl ester M! -isoxazol-5-ylmethylhydrazinecarboxylic acid 60 A mixture of 5-bromomethylisoxazole (1.62g, 10mmol), tert-butyl carbazate (5.29g, 4O0mmol) and sodium carbonate (2.76g, 20mmol) in DMF (25ml) was heated to 8023 in an atmosphere of nitrogen during the day. The mixture was cooled and partitioned between ethyl acetate (10 mL) and water (200 mL). The organic layer is washed with brine (3 x 50 ml) and dried over Na2O). The solvent is removed by ventrative evaporation. Residues of DMF and tert-butylcarbazate are removed by vacuum distillation (5 Ωm of mercury, 80 C). The residue was chromatographed 65 (silica gel, ethyl acetate/hexane (1:1)) to give the final compound as a white solid (1.01g, 4690).

"H NMR (300 MHz, DMSO-a): δ 1.38 (z, 9H), 4.01 (c, 2H); 5.13 (rv, 1H), 6.38 (c, 1H); 8, 38 (5, 1 H); 8.48 (in, 1H).

M -7-chloro-4-hydroxy-2-(pyrrolidin-1-carbonyl)quinoline-3-carboxyl|-M'-isoxazol-5-ylmethylhydrazinecarboxylic acid tert-butyl ester

To a stirred suspension of 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)hydroquinoline-3-carboxylic acid of Example 1 (1.51g, 4./mmol) in THF (50ml) add CMK (4.24g, TOmmol) and the reaction stir for 5 minutes. Add a solution of tert-butyl ester M! -isoxazol-5-ylmethylhydrazinecarboxylic acid (1.0 g, 4.7 mmol) and DMAP (0.06 g,

O.bmmol in THF (1O0ml). The mixture is heated under a reflux condenser, then kept at room temperature for 1 hour. The solid substance is filtered off and washed with DCM (2x B5Oml). The combined filtrate 70 is evaporated to dryness by centrifugation. The solid residue was chromatographed (silica gel, methanol/dx M (1:9)) to give the final compound as an off-white foam (2.09g, 8690). MO (SI) t/2 514/516. 7-chloro-4-hydroxy-2-(5-isoxaline)methyl-1,2,5,10-tetrahydropyridazine(i4,5-B)quinoline-1,10-dione

To a solution of tert-butyl ester of M'-(7-chloro-4-hydroxy-2-(pyrrolidine-1-carbonyl)-quinoline-3-carboxyl|-M'-isoxazol-5-ylmethylhydrazinecarboxylic acid in THF (5 Oml) is added methanesulfonic acid (5.2ml). After 72 18h, the solution is removed by evaporation and the product is precipitated by adding water (100ml). The solid is collected by vacuum filtration, washed with water (2 x 5Oml), dried under vacuum (50Om Hg, 302), suspended in diethyl ether (45 mL) and methanol (b mL) and sonicated (1 h). The solid was collected by vacuum filtration, washed with diethyl ether (2 x 30 mL) and dried under vacuum (500 μm Hg, 302) for 18 h. This gives the final compound as a yellow solid (0.54 g, 81905). 7 "IN NMR (300 MHz, DMSO-dv): δ 5.27 (8, 2M); 6.44 (v, 1H), 7.45 ( ad, 1H, to - 8.7Hz, Ut - 1.8Hz), 8.03 (9, 1H, Ot - 1.8Gu); 8.15 (a, 1H, to - 8.7Hz); 8.53 (8, 1H, U - 1.8Hz); 11.9 (in, 1H), 12.82 (5, 1H).

According to the calculation for C45NoSIMAOde0.4NhO: C, 51.20; H, 2.81; M, 15.92.

Obtained: C, 51.48-51.33; H, 2.79; M, 15.60-15.57. with

Example 8. 7-chloro-4-hydroxy-2-(pyrimidin-2-ylmethyl)-1,2,5,10-tetrahydro-pyridazine-|(4,5-Biquinoline-1,10-dione o

This compound is synthesized as in Example 7, using as a starting compound 4-chloromethylpyrimidine, which is obtained as described in Vagpez.).N. ei ai., Eig. 9. Med. Spent Sleep TNeg., 1988, 23, 211-216. "IN NMR (300 MHz, DMSO-av): 5 12.77 (Bbg 8, 1H); 9.12 (9, IN, y - 1.2Hz), 8.74 (a, 1H, y - 5.4Hz ); 8.05 (a, 1nH,

In - 1.8Hz); 7.45 (aa, tn, y - 21gu, 8.7Gu); 7.40 (а, 1Н, У - 51); 5.20 (c, 2H). -

Example 9. 7-chloro-4-hydroxy-2-(furan-2-ylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-5)quinoline-1,10-dione

IM-(1-aza-2-(2-furanyl)vinyl)tert-butoxy)carboxamide o

2-Furaldehyde (91.9g; "9

O.95mol) and then stand for 2.5 hours. under reflux and cooled to room temperature.

The resulting light brown solid is filtered, dried and used further without further purification. h- "H NMR (300 MHz, DMSO-a): 5 1.52 (b, 9H), 6.45 (ad, 1H, U - 3.33GHz, 1.2Hz); 6.71 (a, 1H, Y - 3.3HC); 7.91 (z, 9H). (tert-butoxy)-M-Kfuranylmethyl)amino|carboxamide

A one-liter three-necked round-bottomed flask is equipped with an additional nozzle, an inlet for nitrogen and a mechanical stirrer. The apparatus is dried in a vacuum and blown with nitrogen. Lithium aluminum hydride (7.75g, 02Omol) and THF (ZOml) are added to the flask. M-(1-aZa-2-(2-furanyl)vinyl)/(tert-butoxy)carboxamide (20g, 0.095mol) was dissolved in THF (2500ml) and slowly over 30 minutes. lithium aluminum hydride is added to the mixed suspension. "» The material remaining in the nozzle is washed into a flask with THF (2 x ZOml). The reaction is stirred overnight, " cooled in an ice bath and carefully quenched with a saturated aqueous solution of Ma 550.5. The resulting mixture is filtered and the filtrate is washed with THF. The combined filtrate and washings are concentrated to oil, which is then stirred for 18 hours. with hexanes (approx. bbOml). The mixture is filtered, the filtrate is concentrated and the desired compound is obtained as a yellow oil (10.0Og, 50965). o TN nMR (300 MHz, DMSO-iv): δ 1.46 (in, 9M), 3.99 (9, 2H, in - 4.9Hz); 4.21 (rg 85, 1H), 6.17 (riv, 1H), o 6.24 (a, tn, U - 3.0 Hz); 6.32 (aa, 1nH, y - 3.0Gu, 1.2Hz). (19-)-N-tert-butoxy)carbonylamino)|7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl)|-M-(2-furanylm (95) 50 ethyl)carboxamide that To mixed to a suspension of 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)hydroquinoline-3-carboxylic acid of Example 1 (26.99g, 84.3mmol) in THF (1300ml) add diisopropylcarbodiimide (13.94g, 110mmol). After stirring for 10 minutes, a solution of (tert-butoxy)-N-(2-furanylmethyl)amino)carboxamide (22.97, 10 mmol) in THF (200 ml) was added dropwise to the reaction mixture. The mixture is stirred for 59 18 hours, then concentrated to a brown resin, which is triturated with chloroform, filtered, washed

HF) with chloroform and dried. This material (approx. 30 g) is used further without purification. 7 7-chloro-4-hydroxy-2-(2-furanylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-b)quinoline-1,10-dione

To a solution of (19-)-M-tert-butoxy)carbonylamino)|7-chloro-4-oxo-2-(pyrrolidinyl-carbonyl)(3-hydroquinolyl)|1-M-(2-furanylmethyl)carboxamide ( 29.00 g, 56 b, mmol) in THF (1000 ml) add methanesulfonic acid (118 ml) and stir the reaction for 18 hours. at room temperature, the mixture is poured into Zl of ice water and filtered to obtain a whitish solid, which is dissolved in hot THF (approx. Tl) and then concentrated to half the volume.

The resulting suspension is poured into 2 liters of ice water and after 20 minutes. filter After drying this material, the final compound is obtained as a white solid (144g, 7490), temp. sq. 2265260. b TN nMR (300 MHz, DMSO-iv): δ 5.08 (c, 2M); 6.37 (4, yin, 9 - З0Hz), 6.42 (da, yin, y - З3,ОГHz, 1.5Гу),

7.43 (a, 1H, 2 - 7.8Hz), 7.54 (c, 1H), 8.00 (j, 1H, Y - 1.5Hz); 8.14 (а, 1Н, to - 8, Hz).

According to the calculation for Si6NoSIMaOu: C, 55.91; H, 2.93; M, 12,23.

Obtained: C, 56.10; H, 2.98; M, 12.03.

Example 10. 7-chloro-4-hydroxy-2-(furan-3-ylmethyl)-1,2,5,10-tetrahydropyridazine-|4,5-b)quinoline-1,10-dione

M-(1-aza-2-(3-furyl)vinyl)x/tert-butoxy)carboxamide

To a solution of furan-3-carboxaldehyde (2.0g, 2immol) add tert-butyl carbazate (2.8g, 21mmol) and conc.

NOI (5 drops). The solution is stirred frequently. and concentrate. The resulting solid was triturated with hexanes to give the final compound (4.2g, 20mmol, 9795) as a peach-colored solid.

TN NMR (300 MHz, DMSO-av): 5 10.77 (Bg 5, 1H), 8.02 (in, 1H), 7.94 (in, 1H), 7.71 (ad, 1H, 9 - 1 .5GHz, 1.5Hz); 6.77 (9, 1H, U - 1.5Gu); 1.45 (in, 9H). (tert-butoxy)-M-((3-furylmethyl)amino|carboxamide

Sodium cyanoborohydride (3.0 g, 4 Avmmol) and acetic acid (bml). The solution is heated to 659 for 75 hours, then cooled to room temperature, quenched with water and methanol removed in vacuo. The residue is taken up in ethyl acetate and washed with saturated sodium bicarbonate, water and brine and dried over

Ma»zO), then filtered and concentrated. The residue was purified by silica gel chromatography (hexanes/ethyl acetate (4:1)) to give the final compound (90 mg, 4.4 mmol, 4695) as a clear oil.

TN NMR (300 MHz, DMSO-av): 5 8.26 (rg 8, 1H), 7.58 (ad, 1H, y - 1.5 GHz, 1.5 Hz), 7.53 (v, 1H), 6 ,42 (d, 1H, y - 1.2Gu); 4.64 (rgv, 1H), 3.69 (in, 2H); 1.39 (in, 9H). (tert-butoxy)-M-1(7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl)|-M-3-"-furylmethyl)carbonylamino)carboxamide

CMK (3.7g, 8.6mmol) was added to a stirred suspension of 7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)-hydroquinoline-3-carboxylic acid of Example 1 (14g, 4.4mmol) in THF (25ml). To this bright yellow mixture was added a solution of c (tert-butoxy)-N-3-furylmethyl)amino)carboxamide (93 Ωg, 4.4 mmol) and M,M-dimethylaminopyridine (8 Ωg, 0 bbommol) in THF (20 mL) with stirring. The mixture is maintained under a reflux condenser in a nitrogen atmosphere according to, then cooled and filtered. The filtrate is concentrated and a yellow solid foam is obtained, which is chromatographed on silica gel (methanol(1095)/СНо.сСИ») and the final compound is obtained in the form of a pale yellow solid, which is used directly in the future. -- 7-chloro-4-hydroxy-2-(furan-33-ylmethyl)-1,2,5,10-tetrahydropyridazine4,5-)quinoline-1,10-dione

To a mixture of (tert-butoxy)-M-117-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl)|-M-3-"furylmethyl)carbonylamino)carboxy and samide (14g, 2.8mmol) in A solution of methanesulfonic acid CO (7.2 ml, 110 mmol) in THF (25 ml) is added to THF (30 ml) at room temperature. The solution is stirred overnight while adding water to stir

From deposition. The solid substance is collected, washed with water and diethyl ether and treated with ultrasound (15 min.) - in methanol (1095) in diethyl ether solution. The resulting yellow solid was collected, washed with diethyl ether and dried at 309 and 5 µm Hg. Art. during the Agreement and obtain the final substance (75Omg, 2.1 mmol, 75905) as a pale yellow solid. " "IN NMR (300 MHz, DMSO-av): δ 12.63 (Bg 8, 1H), 11.91 (v, 1H); 8.13 (a, 1H, U - 8.7 GHz), 8.02 (a, 1H, y - 1.5Gu); C is 7.65 (in, 1H), 7.61 (a, 1H, y - 1.5Hz); 7.43 (da, 1H, У - 1, 8Hz, 8.7Gu), 6.46 (in, 1Н); 4.92 (in, 2Н). s According to the calculation for С46НлоМа3О4Сив 0.1Н20: С, 55.62; Н, 2.98; M, 12, 16. :z» Obtained: C, 55.67; H, 3.15; M, 11.77.

Example 11. 7-chloro-4-hydroxy-2-(thien-3-ylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-B)quinoline-1,10-dione

This compound is synthesized as in Example 4, using thiophene-2-carboxaldehyde as a starting material. -I "IN NMR (300 MHz, DMSO-dv): δ 12.71 (rgv, 1H), 11.91 (in, 1H); 8.14 (9, 1H, in - 8.7GHz), 8.02 (c, 1H); 7.42-7.45 (t, 2H); 7.10 (a, 1H, y - 3.0Hz); 6.98 (ad, 1H, y - 5.1Hz), 5 ,23 (c, 2H). o Example 12. 7-chloro-4-hydroxy-2-(thien-3-ylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-B)quinoline-1, 10-dione

Ge) This compound is synthesized as in Example 4, using thiophene-2-carboxaldehyde as a starting material. syu 50 "NN NMR (300 MHz, DMSO-iv): 5 12.64 (rg 5, 1H), 11.92 (rg 5, 1H); 8.14 (9, 1H, y - 8.7 Hz), 8 .02 (a, 1H, yt 1.5Hz); 7.48-7.54 (t, 1H); 7.43 (aa, 1H, U - 1.2Gu, 8.7Hz), 7.37 ( c, 1H), 7.08 (a, 1H, Y - 4.5HC), 5.08 (c, 2H). -. and Example 13. 7-chloro-4-hydroxy-2-(benzo|b| hyen-3-ylmethyl)-1,2,5,10-tetrahydro-pyridazine(i4,5-B)quinoline-1,10-dione

Benzo|bigiophene-2-carbaldehyde

5ml of 1.6M p-butyllithium in hexanes was added to a solution of benzo|bithiophene (10g, 74.5mmol) in dry THF (12ml) at -782C and after 100 minutes. add DMF (2 mL, 298 mmol). The reaction is heated to room temperature and kept at under reflux. The THF is evaporated, and the residue is poured into 1M MEDIA with ice.

The solution is extracted with diethyl ether (twice), the combined ether extract is washed with 1 M NaCl (three times), saturated with Manso's brine and brine, and dried with NaCl, which is then filtered. The filtrate is concentrated to oil, which is treated with MansSoO with. The formed solid is dried over M950. and evaporation gave the final compound 60 as a yellow oil (3.2 g, 2690). "YAN NMR (Z00MHZc, SOSIv): 5 7.44 (aa, yin, y - bogtz, 7.2Hz), 7.51 (aa, yin, U - 6.oHz, 8.1Hz); 7.91 ( a, many

U - 8.1Gu), 7.95 (9, 1H, U - 7.8Hz), 8.04 (v, 1H), 10.12 (v, 1H). (tert-butoxy)-M-1-aza-2-benzo|B|hyen-2-ylvinyl)/carboxamide

To a solution of benzo|B)thiophene-2-carbaldehyde (3.2 g, 19.7 mmol) and tert-butyl carbazate (2.6 g, 19.7 mmol) in ethanol (20 ml) add 3 drops of concentrated HCl. Through ZOkhvil. the mixture is filtered, the solid is washed with diethyl ether, and after drying in a vacuum, the final compound is obtained (Z, 8 g, 70905).

TN nMR (Z0OMHC, SOSIv): δ 1.54 (v, 9H), 7.30-7.36 (t, 1H); 7.40 (in, 1H), 7.71-7.74 (t, 1H), 7.89 (in/1

H), 8.31 (Bg 5.1H). (tert-butoxy)-M-((benzo|b|hyen-2-ylmethyl)amino|carboxamide

To a suspension of (tert-butoxy)-M-1-aza-2-benzo|B)hyen-2-ylvinyl)ucarboxamide (1.8g, b,bmmol) in THF (bml) sodium cyanoborohydride (0.75g, 11 .9 mmol). A solution of p-toluenesulfonic acid (1.86 g, 9 U, mmol) in THF (bml) was added dropwise. After stirring overnight, the reaction was diluted with ethyl acetate and washed with saturated Manso's and saturated Massey's. The ethyl acetate solution is dried over CoCO3, which is then filtered at 70 V and the filtrate is concentrated under reduced pressure. The resulting solid is treated with saturated NaHSO 3 overnight and extracted with DCM. The DCM solution was dried over Ma 250.4, which was then filtered, and the filtrate was concentrated to give the final compound as a white solid (1.7g, 7696). "H NMR (Z00MHZ, SOSI"): δ 1.44 (z, 9H), 4.68 (v, 2H), 7.36-7.45 (t, 2H); 7.45 (v, 1H) , 7.78-7.86 (t, 2H).

Tert-butyl ester

N-Carboxylic Acid

CMK (3.19 g, 3.19 g, 7.Omol) and then a solution of (tert-butoxy)-M-((benzo|b|hien-2-ylmethyl)amino)carboxamide (1.37g, 5.00mmol) and dimethylaminopyridine (27.8g, 0, 21 mmol) in THF (15 ml). The reaction is heated overnight under reflux and the mixture is filtered.

The concentrated filtrate was purified by chromatography (Meon/SnH2Cl - 5:95 (v/v)) on silica gel to give the final compound as a yellow solid (771mg, 24905). 7-chloro-4-hydroxy-2-(benzo|b)hyen-3-ylmethyl)-1,2,5,10-tetrahydro-pyridazine(4,5-biquinoline-1,10-dione c

To the mixed mixture of tert-butyl ester (9

M'-Kbenzo|b)hyen-2-ylmethyl)-M'-H7-chloro-4-oxo-2-(pyrrolidine-1-carbonyl)-1,4-dihydroquinoline-3-carbonyl|-hydrase of ncarboxylic acid ( 7bOmg, 1.3mmol) and dry THF (1bml) at 09Сb in a nitrogen atmosphere add methanesulfonic acid (3.Oml, 4.44g, 46.2mmol). After stirring overnight, the TG/F was evaporated and the residue was cooled in an ice bath. Add water and collect the precipitate, sonicate in methanol, -- dry in vacuum to obtain the final compound in the form of a whitish solid (47Omg, 8895), temp. with sq. 30096,

TN NMR (300 MHz, DMSO-iv): δ 5.38 (in, 2M); 7.30-7.38 (t, 2H), 7.40 (c, 1H), 7.44 (y, 1H, 9 - 8.7Gu); at 7.50 (a, tn, U 7.2Hz); 7.82 (a, tn, U - 7.2Hz); 7.82 (а, 1Н, У - 7.2Hz); 8.06 (c, 1H), 8.15 (a, 1H, U - 8.7Hz). co

According to the calculation for SooH4i2SIMz3Ozz: C, 58.61; H, 2.95; M, 10.25.

Obtained: C, 58.42; H, 3.19; M, 10,20. -

Example 14. 7-chloro-4-hydroxy-2-(1,3-thiazol-2-ylmethyl)-1,2,5,10-tetrahydropyridazine-|4,5-B)quinoline-1,10-dione ( Tert-butoxy)-M-((benzo|bhien-2-ylmethyl)amino|carboxamide

To a solution of thiazole-2-carbaldehyde (0.95g, 8.42mmol) and tert-butyl carbazate (1.17g, 8.87mmol) in ethanol (15ml) add 1.10ml of glacial acetic acid and then sodium cyanoborohydride (2.18g , 34.7 mmol). The reaction mixture with 70 is heated to 5092 and stirred for 72 hours. The reaction was quenched with 2M Mahon (30 ml) and the solution was extracted with ethyl acetate (3 x 30 ml). The combined organic layers were washed with brine (40 mL) and dried over Na2SO4, which was then filtered and the filtrate purified by chromatography (ethyl acetate/xM (25:75)) on silica gel to give the final compound as an off-white solid (0, 62g, 32905).

TN nMR (300 MHz, DMSO-dv): δ 1.38 (in, 9M); 4.15 (а, 2Н, 2 - 3.9Hz); 5.34 (9, yin, y - 3.9Hz); 7.63 (49, - and 1H, U - 3.3Hz); 7.70 (а, 1Н, У - 3.3Hz); 8.42 (bBg 8, 1H).

M-C-tert-butoxy)carbonylamino|7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)(3-hydroquinolyl)|-M-(1,3-thiazol-2-ylmoethyl)carboxamide so To a stirred mixture of 7- of chloro-4-oxo-2-(pyrrolidinylcarbonyl)hydroquinoline-3-carboxylic acid of Example 1 (0.90g, 2.80mmol) and dry THF (bOml) in a nitrogen atmosphere add CMK (1.45g, 3.42mmol) and then a solution of (tert-butoxy)-M-(1,3-thiazol-2-ylmethyl)amino|carboxamide (0.5g, 2.20mmol) and dimethylaminopyridine 7"79.1g, - M O,ppmmol) in THF ( 15ml). The reaction mixture is heated under reflux overnight and after cooling is filtered. The concentrated filtrate is purified by chromatography (Meon/snH Si" - 5:95 (v/v)) on silica gel to give the final compound as a yellow solid substances (0.97 g, 8390). 10-dione

To a stirred mixture of (iF) M-tert-butoxy)carbonylamino|7-chloro-4-oxo-2-(pyrrolidinyl-carbonyl)(3-hydroquinolyl)|-M-(1,3-thiazol-2-ylmethyl) of carboxamide (7 mg, 1.3 mmol) and dry THF at 02С in a nitrogen atmosphere, methanesulfonic acid (5.4 ml, 7.99 g, 83.2 mmol) is added. After stirring overnight, the THF was evaporated and the residue was cooled in an ice bath at 60°C. Water is added and the precipitate is collected and sonicated in methanol. The solid substance is collected by suction filtration, dried in vacuum. The final compound is obtained as a whitish solid substance (648mg, 7895), temp. sq. "30090.

TN nMR (300 MHz, DMSO-iv): δ 5.38 (z, 2M); 7.45 (4, yin, y - 8.7Hz); 7.90 (a, yin, 9 - 8.7Hz); 7.70 (49, 1H, U - 3.3Hz); 7.76 (9, 1H, y - 3.9Hz); 8.04 (5, 1H), 8.15 (9, 1H, U - 8.7Hz). because According to the calculation for C45NioSIM3OZe NaONZSZO»N: C, 40.38; H, 3.39; M, 11.77.

Obtained: C, 40.63; H, 2.98; M, 11.39.

Example 15. 7-chloro-4-hydroxy-2-(imidazol-2-ylmethyl)-1,2,5,10-tetrahydropyridazine-|4,5-8|quinoline-1,10-dione

IM-(1-aza-2-imidazol-2-ylvinyl)/(tert-butoxy)carboxamide

To a solution of 2-imidazolecarboxaldehyde (10.2 g, 10 mmol) add tert-butyl carbazate (2.8 g, 21 mmol) and conc.

NOAH The solution was stirred overnight, concentrated and triturated with hexanes to give the final compound (22g, 99905) as a white solid.

TN NMR (300 MHz, DMSO-α): δ 1.47 (z, 9M), 7.08 (5, 2H), 7.92 (c, 1H), 10.93 (rg 5, 1H), 12, 58 (Bgv, 1N). (tert-butoxy)-N-Cimidazol-2-ylmethyl)amino|carboxamide

A mixture of 1095 palladium on carbon and M-(1-aza-2-imidazol-2-ylvinyl)ut-butoxy)-carboxamide (3.Og, 14 mmol) in methanol and concentrated HCl (1.15 ml, 14.0 mmol) hydrogenate (40 lb/sq. in. - 2.bat) at room temperature for 18 hours. The reaction is filtered through diatomaceous earth, the filtrate is evaporated under reduced pressure and an oil is obtained, which is neutralized by adding sodium hydroxide (5M, 2.8ml) and diluted with ethyl acetate. The ethyl acetate layer is washed with water (20 ml) and aqueous saturated sodium chloride (20 ml) and dried over NaCl. Ethyl acetate was removed to give the final compound as an oil (2.42 g, 80965).

TN NMR (300 MHz, DMSO-dv): δ 1.36 (in, 9M); 3.84 (a, 1H, y - 4.2Hz); 4.82 (rg 8, 1H), 6.91 (z, 2H), 8.25 (rg 5, 1H), 11.84 (Bg in, 1H).

N-tert-butoxy)carbonylamino!7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)-(hydroquinolyl)-N-(imidazol-2-ylmethyl)carboxamide

A mixture of //7-chloro-4-oxo-2-(pyrrolidinylcarbonyl)hydroxyquinoline-3-carboxylic acid of Example 1 (2.42g, 7.54mmol), (tert-butoxy)-N-(imidazol-2-ylmethyl)amino |-carboxamide (2.0g, 94Zmmol) in dry THF (50ml) is refluxed for 18 hours. The reaction is filtered, the solid is collected and washed with water and then with diethyl ether. This material was dried in vacuo to give the final compound as a white solid (1.0 g, 2590). c 7-chloro-4-hydroxy-2-(imidazol-2-ylmethyl)-1,2,5,10-tetrahydropyridazine-|4,5-)quinoline-1,10-dione o

To the solution

M-Ct-butoxy)carbonylamino| 7-Chloro-4-oxo-2-(pyrrolidinylcarbonyl)-(hydroquinolyl|-N-(imidazol-2-ylmethyl)carboxamide (1.0g, 1.p94mmol) in THF (30ml) was added methanesulfonic acid (bml) and the reaction is stirred overnight. The volatile components are removed under vacuum and diethyl ether (d0O0ml) is added to the oily residue. The mixture is stirred for 10 minutes and left to form two layers - an ether layer and a brown oil layer. The ether is decanted, and water is added to the brown oil ( After a short time, a precipitate forms, which is collected by vacuum filtration, washed with diethyl ether (Х20 ml) and treated with ultrasound (15 min.) in a mixture of diethyl ether and methyl alcohol (10:1). The material is filtered, washed with diethyl ether, and dried in a vacuum and co obtain the final compound (0.24 g, 25905).

Zo "IN NMR (300 MHz, DMSO-dv): 5 2.31 (SNUZOZN); 5.41 (in, 2H), 7.42 (a, 1H, 9 - 8.7 Hz); 7.67 (in, 2H); 7.99 in (8, 1H); 8.15 (a, 1H, U - 8.7Hz); 12.7 (Bg in, 1H), 11.98 (rg 5), 12.93 ( rg c, 1H), 14.28 (rgv, 1H).

According to the calculation for C45NoSIMyOze1.6SNaUSZOZN: C, 40.08; H, 3.32; M, 14.08.

Obtained: C, 40.37; H, 3.12; M, 14.34. « du Test of biological functions - with Test A: Inhibition of binding |M|MY 105,519

The binding of compounds to the glycine site of the MOMA receptor can be assessed by measuring the ability of the compound to inhibit the binding of tritiated MOI 105.519 to brain membranes that have this receptor.

The rat brain membranes used in the experiments were obtained from Apaimiis! Viioiodisai

Zeguisev Ips and were prepared according to the method of V.M. Vagop ei ai., U. RNagtasoi. Ex. Thiep, 250, 162 -1 (1989). Fresh brain tissue from Zrgadoe-Oamieu rats, including cerebral cortex and hippocampus, was homogenized in 0.32 M sucrose and centrifuged at low speed to separate cell membranes from other components of cells. Membranes were washed three times with deionized water and then treated with 0.0495 µM Tiop X-100, after which they were washed twice in 500 mM Ti3-ditrate buffer, pH 7, and frozen at -809C for further

Using. about INIMO 105.519 was purchased from Ategepat, cold MY 105.519 - from Vidt/VVI. Binding analysis - performed according to V.M. Vagop ei a/., 9). Rpagtasoi Ex. Tpep, 279, 62 (1996), as described below.

On the day of the experiment, the brain membranes were thawed and suspended at room temperature in X0OMM hys-acetate buffer, pH 7.4 ("TAV"). Protein (75 μg/ml) with VioKid dye was used for competitive binding. The experiments were carried out on 96-socket boards. Membranes were incubated with 20 μl of compounds of different concentrations and 1.2 nM I(ZMINOI 105.519 during incubation at room temperature in a volume of 250 μl. o Nonspecific binding was determined using 100 μM of unlabeled MY 105.519. Unlabeled MY 105.519 and ime) compounds were dissolved as basic 12.5 mM solutions in DMSO. The final concentration of DMSO in each well was kept below 195 as it was found that this concentration did not affect the binding results. After incubation, unbound | REMOVABLE 105,519 was removed by filtering the CE/V Opiykeg board using the RasKag collector. Filters were washed 4 times with ice-cold TAV (full amount of buffer 1.2 ml).

The plates were dried overnight at room temperature and the bound radioactivity was measured with a RuskKag counter

TorsSoishpi after adding 45 μl to each nest of MISKOZSIMT O.

Human brain membranes were obtained from Apaiuiis! Vioiodisa! Zegmisez Ips. and were prepared as described 65 for rat membranes.

Data analysis was performed using the software packages of Misgozoia Eheya! and SgarpRai Rgig2t, and the potency of the compounds was measured in Ki (nm).

Test B. Formalin test

The formalin test is used to evaluate the ability of a compound to inhibit formalin-induced nociceptive behavior in rats (O. YuobBriivvop ei ai., Raip 4, 161-174 (1977); N. UMUpeiiIeg-Asef ei ai.,

Resusporpagptasoiodu 104, 35-44 (1991); T.). Sodete ei aiYi., Raip 54, 43-50 (1993). In this test, two distinct phases of formalin-induced behavior are observed. The first phase of the response, caused by acute nociception in response to the injection of a caustic chemical (formalin) into the paw, lasts 0-2 min. After injection within 5-15 minutes. there is a period of appeasement. After that, after 15 minutes. begins and continues to bohvil. /o the second phase of the reaction, caused by the sensitization of central neurons in the dorsal horn. Sensitization of central spinal neurons enhances the afferent noxious input and causes the transmission of increased pain to the brain. Therefore, inhibition of the second phase of the reaction determines the central mechanism of drug action.

The formalin test procedure is as follows: male rats are placed in a Plexiglas chamber and observed for 30-45 minutes. the main features of their behavior. Animals are injected with a vehicle (control group) or different doses of the tested compound in three hours. before injection of O0O05 ml of sterile 195th formalin under the dorsal skin of the hind paw.

The number of paw twitches (reactions) during the first (0-5 min.) and second (20-Zbhmin.) phases was counted and recorded. The nociceptive response was compared with the average corresponding number in the control group (saline) and calculated as inhibition in 95956. EHUOvo is the dose of the compound that gives 5095-no inhibition of the nociceptive response in the first or second phase. The first phase of the reaction can be inhibited by compounds that act peripherally and compounds that act centrally. The second phase of the reaction can be inhibited by compounds that have a central effect. inhibition of nociceptive reaction (90) - 100 x (number of reactions in the sexual intercourse group)/ (number of reactions in the control group)

The Student's test was used to determine the statistical significance of the effect of the compound. Data are given in 9090 inhibition. with

Test C Model of neuropathic pain (Chronic Compression)

The antihyperalgesic qualities of the compound can be tested on the Chronic Compression Model (CCM). The test is a model of neuropathic pain associated with nerve damage that results directly from trauma or compression or is an indirect consequence of diseases such as infection, cancer, metabolic disorders, toxins, malnutrition, immunological dysfunction, and musculoskeletal changes. In this model, unilateral peripheral "-- hyperalgesia in rats is created by applying a ligature to the nerve (5.). Veppei ei a!., Raip 33, 87-107 (1988)).

According to the procedure, Zrgadoe-OaumlLeu rats (250-350g) are anesthetized with sodium pentobarbital and exposed and. common sciatic nerve at the level of the middle of the thigh by an incision through Biserz Tetogiv. A part of the nerve (approx. 7 mm) is freed from the tissues and ligatured with chrome catgut in four places with an interval of mm. The incision is closed in layers and the animals are left to recover. Thermal hyperalgesia is measured by the paw withdrawal test (K. Nagogeamez ei a!., Raip 32, 77-88 (1988)). Before the test, the animals are accustomed to eating on a raised glass floor. The heat beam from the source is directed to the area of the sciatic nerve on the hind paw through the glass floor in 20s periods to avoid skin injury. Latencies of the withdrawal reflex in both hind paws are measured.

Injured paws with nerve ligatures show shorter paw withdrawal latencies compared to " 70 Uninjured paws. Responses to the test compounds were assessed at various times after oral administration to determine the onset and duration of action of the compound. When performing the test, groups of SCI rats received the vehicle and or the compound orally three times a day for 5 days. Paw withdrawal latencies were measured every day for "" 10 minutes. before and after 2 or Accord. after the first daily dose. The effectiveness of the compound was evaluated as the average decrease in

Fdodo of hyperalgesia compared to the control group of animals as 100 x (mean in the control group - mean in the compound group)/ (mean in the control group) -I Data analysis was performed through the test of comparison of multiple means (Data-ta test) and its results and the potencies of the compounds were presented as MED (minimum effective dose) in mg/kg/day, which determines a statistically significant reduction (in 90705) of hyperalgesia.

OO Tab. 1 contains the results of tests A, B and C for some compounds of the invention. Lack of data in the table. means that the test was not performed. (95) - From Table 1

Test A Test B Test C Test C

Ki (nm) First phase Dose (95 inhibitions) Second phase Dose (95 inhibitions) MED (95 inhibitions)

Experiment 1 73 70 (5295) 70 (5795) 30 (7195) 59 Experiment 2 99 200 (7496) 200 (7196) 30 (5995) (F) Experiment C 146 70 (5695) 200 (4695) 30 (3195) ka Experiment 4 4A 30 (4495)

Experiment 5 371 in Experiment 6 189 30 (6495)

Experiment 7 39 15 (4995)

Experiment 8,513

Experiment C 61 200 (5696) 200 (46965) 15 (4995)

Experiment 10 24 30 (4195) 65 Experiment 11 29 15(-1795)

Experiment 12 34 15(1795)

Experiment 13 56 15(195)

Experiment 14 128 30 (2695)

Experiment 15 103

Claims (5)

The formula of the invention 1. The method of treating a patient suffering from pain, which consists in the introduction of an effective pain-relieving amount of any compound of the structural formula I: on o and A with MN in M sht II M -E on deA - (СНо)д, ап -0,1,2, Zabod, and Or - 5- or b--linked heteroaryl group or its benzo derivative, which has 1, 2 or 3 oxygen or nitrogen atoms, or sulfur in the ring, and B - halogen. 2. The method according to claim 1, which is characterized by the fact that it consists in the introduction of an effective pain-relieving amount of a compound of the structural formula I, in which c O is selected from the group consisting of pyridyl, quinolyl, pyrazinyl, pyridizinyl, furanyl, benz(Ir|furanyl, o imidazolyl, oxazolyl, thienyl, benz|(B)hienyl and thiazolyl. Q. The method according to claim 1, which differs in that it consists in the introduction of an effective pain-relieving amount of the compound of the structural formula II: he o I. - She ZM i DI Ge) mya s M o si o on m. 4. The method according to item C, which differs in that it consists in the introduction of an effective pain-relieving amount of a compound of the structural formula II, where O is selected from the group consisting of pyridyl, quinolyl, pyrazinyl, pyridizinyl, furanyl, benz(Ir|furanyl, imidazolyl, oxazolyl, thienyl, benz|(B)hienyl and thiazolyl. 5. The method according to item 3, which differs in that it consists in the introduction of an effective pain-relieving amount of a compound selected from the group formed by 7-chloro-4-hydroxy-2-(4-pyridylmethyl)-1, 2,5,10-tetrahydropyridazine4,5-B)quinoline-1,10-dione, "7-chloro-4-hydroxy-2-(3-pyridylmethyl)-1,2,5,10-tetrahydropyridazine4,5-B )quinoline-1,10-dione, 7-chloro-4-hydroxy-2-(2-pyridylmethyl)-1,2,5,10-tetrahydropyridazine4,5-B)quinoline-1,10-dione, 7-chloro -4-hydroxy-2-benzo|a4|furan-2-ylmethyl-1,2,5,10-tetrahydropyridazine|(4,5-B)quinoline-1,10-dione, -and 7-chloro-4- hydroxy-2-(quinolin-4-ylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-B)quinoline-1,10-dione, o 7-chloro-4-dimethylcarbamoyl-2-pyridine-4 -ylmethyl)-1,2,5,10-tetrahydropyridazine|(4,5-B|quinoline-1,10-dione, 7-chloro-4-hydroxy-2-(pyrazin-2-ylmethyl)-1,2 ,5,10-tetrahydropyridazine(I4,5-fSh)quinoline-1,10-dione, (65) 7-chloro-4-hydroxy-2-(5-isoxazolino)methyl-1,2,5,10-tetrahydropyridazine (4,5-quinoline-1,10-dione, about 20 7-chloro-4-hydroxy-2-(pyrimidin-2-ylmethyl)-1,2,5,10-tetrahydropyridase n(4,5-51-quinoline-1,10-dione, 7-chloro-4-hydroxy-2-(furan-2-ylmethyl)-1,2,5,10-tetrahydropyridazine(I4,5-Shiquinoline- 1,10-dione, "6 7-chloro-4-hydroxy-2-(3-furylmethyl)-1,2,5,10-tetrahydropyridazine (4,5-quinoline-1,10-dione, 7-chloro- 4-hydroxy-2-(thien-2-ylmethyl)-1,2,5,10-tetrahydropyridazine (4,5-quinoline-1,10-dione, 7-chloro-4-hydroxy-2-(thien-3 -ylmethyl)-1,2,5,10-tetrahydropyridazine (4,5-quinoline-1,10-dione, 22 7-chloro-4-hydroxy-2-(benzo|b)hyen-2-ylmethyl)-1 ,2,5,10-tetrahydropyridazine4,5-B)quinoline-1,10-dione, Ge! 7-chloro-4-hydroxy-2-(1,3-thiazol-2-ylmethyl)-1,2,5,10-tetrahydropyridazine4,5-c)quinoline-1,10-dione: and 7-chloro- 4-Hydroxy-2-(imidazol-2-ylmethyl)-1,2,5,10-tetrahydropyridazine(i4,5-B)quinoline-1,10-dione. where 6. A pharmaceutical composition containing an effective pain-relieving amount of a compound of structural formula Y together with a pharmaceutically acceptable excipient or diluent; 60 b5 he o and in the west MI on de A - (СНО), and n has the value 0, 1, 2, З or 4, Or is a 5- or 6b-linked heteroaryl group or its benzo derivative, which has oxygen or nitrogen or sulfur atoms in the ring 1, 2 or 3, and B is a halogen. Official bulletin "Industrial Property". Book 1 "Inventions, useful models, topographies of integrated microcircuits", 2005, M 7, 15.07.2005. State Department of Intellectual Property of the Ministry of Education and Science of Ukraine. with o "- so so so and - shi s ;" -And (95) (95) at 50 - F) name) 60 b5