WO2002007727A1

WO2002007727A1 - Use of pyridazino(4,5-)indole-1-acetamide derivatives for preparing medicines for treating pathologies related to the dysfunction of peripheral benzodiazepin receptors

Info

Publication number: WO2002007727A1
Application number: PCT/FR2001/002369
Authority: WO
Inventors: Régine BARTSCH-LI; Laurent Dubois; Yannick Evanno; Jacques Froissant; Pascal George; Benoît MARABOUT; Franck Marguet; Mireille Sevrin
Original assignee: Sanofi-Synthelabo
Priority date: 2000-07-24
Filing date: 2001-07-20
Publication date: 2002-01-31
Also published as: AR029960A1; AU2001278549A1; FR2811897A1

Abstract

The invention concerns the use of a compound, in the form of base or addition salt to an acid, corresponding the general formula (I) wherein: X represents a halogen atom; Y represents one or several atoms or groups selected among hydrogen, halogens and hydroxy, methyl and methoxy groups; R1 represents a hydrogen atom or an alkyl group; R2 represents a hydrogen atom, an alkyl group, a hydroxyalkyl group, a cycloalkyl group, a cycloalkyl-alkyl group, a phenyl group, a pyridinyl group or a phenylalkyl group, for preparing a medicine for preventing or treating a disease related to the dysfunction of peripheral benzodiazepin receptors

Description

USE OF PYRIDAZINO (4, 5) INDOLE-1-ACETAMIDE DERIVATIVES FOR THE PREPARATION OF DRUGS FOR THE TREATMENT OF DISORDERS RELATED TO BENZODIAZEPINE PERIPHERAL RECEPTORS

Within the framework of the search for compounds which can promote the regeneration of axons of peripheral nerve cells after injury, a subclass has been identified, among the compounds of international application O-9906406.

10 compounds of general formula (I)

20 X represents a halogen atom,

Y represents one or more atoms or groups chosen from hydrogen, halogens and hydroxy, methyl and ethoxy groups,

Rx represents a hydrogen atom or a group (C ^ C ^ alkyl,

25 R ₂ represents a hydrogen atom, a linear or branched (C- _L -C ^) alkyl group, a hydroxy (C ₁ -C ₄ ) alkyl group, a (C ₃ -C ₇ ) cycloalkyl group, a group (C ₃ -C ₇ ) cycloalkyl (C ^ -Cs) - alkyl, a phenyl group, a pyridinyl group or a phenyl group (C- _L -C ^ alkyl.

30

The compounds of general formula (I) can exist in the form of bases or of addition salts with acids.

These compounds have a strong affinity for peripheral benzodiazepine receptors (p sites, or PBR), and some induce, in particular, a reduction in neuronal loss in the facial nucleus after section of the facial nerve.

Among the compounds of particular interest for the use according to the invention that may be mentioned, for example, 7-chloro-5-methyl-1- [2- (4-methylpiperazin-1-yl) -2-oxoethyl] -4-oxo-3-phenyl -3.5 ~ dihydro-4iï-pyridazino [4, 5-jb] - indole.

The latter can be prepared according to the following procedure, given by way of example. The synthesis of 7-chloro-5-methyl-4-oxo-3-phenyl-3, 5-dihydro-4iï-pyridazino [4, 5-jb] indole-1-acetonitrile is already described in international application PCT / FR00 / 00135.

Example 1 (Compound No. 2 in the table below). 7-chloro-5-methyl-4-oxo-1- [2-oxo-2- (4-methylpiperazin-1-yl) ethyl] -3-phenyl-3, 5-dihydro-4iT-pyridazino hydrochloride [4 , 5-i] indole (1: 1).

1.1. 7-chloro-5-methyl-4-oxo-3-phenyl-3, 5-dihydro ~ 4iT-pyridazino acid [4, 5-b) indole-1-acetic. A solution of 25.5 g (73.1 mmol) of 7-chloro-5-methyl-4-oxo-3-phenyl-3, 5-dihydro-4H-pyridazino is stirred for 4 h at reflux [4, 5 -b] indole-1-acetonitrile in 1.8 l of concentrated hydrochloric acid and 1.8 l of acetic acid. The reaction medium is cooled, filtered and the filtrate is diluted with water. A solid is collected by filtration, washed with water and dried under reduced pressure. It is dissolved in 1.5 l of water and 250 ml of a concentrated ammonia solution. An insoluble material is filtered and the filtrate is acidified with a solution of concentrated hydrochloric acid. A solid is collected by filtration, which is rinsed with water and then with diethyl ether and dried under reduced pressure.

12.1 g (2.62 mmol) of compound are isolated in the form of a white solid. Melting point: 195-197 ° C.

1.2. 7-chloro-5-methyl-4-oxo-1- [2-oxo-2- (4-methylpiperazin-1-yl) ethyl] -3-phenyl-3, 5-dihydro-4iT-pyridazino hydrochloride [4 , 5-jb] indole (1: 1).

Under argon, to a solution of 1.0 g (2.7 mmol) of acid 7-chloro-5-methyl-4-oxo-3-phenyl-3, 5-dihydro-4H-pyridazino_ [4, 5-jb] indole-1-acetic in 80 ml of dichloromethane, 0.90 g ( 5.55 mmol) of 1,1 '-carbonylbis-1H-imidazole and the reaction mixture is stirred for 1 h 30 min at 45 ° C. The mixture is cooled to room temperature and 0.56 g (5.6 mmol) of 1-methylpiperidine in 1 ml of dichloromethane is added and the reaction mixture is stirred for 12 h. 10 ml of water and 150 ml of dichloromethane are added, then, dropwise, concentrated hydrochloric acid to a pH of approximately 3 to 2. The organic phase is decanted, washed with water then with a saturated aqueous solution of sodium hydrogencarbonate. The organic phase is dried with sodium sulfate, the solution is filtered, the filtrate is concentrated under reduced pressure and the residue is purified by chromatography on a column of silica gel. 0.89 g of a white solid is isolated.

The hydrochloride is prepared using 0.89 g (0.002 mole) of base and 3 ml of a solution of hydrochloric acid (about 2M) in methanol. It is recrystallized from a mixture of dichloromethane and methanol.

0.62 g of hydrochloride is isolated in the form of a white solid. Melting point: 240-243 ° C.

Example 2 (Compound No. 1 in the table below).

7-chloro-5-methyl-4-oxo-1- [2-oxo-2- (pipéra_ zin-1-yl) ethyl] -3-phenyl-3, 5-dihydro-4H-pyridazino hydrochloride [4, 5-jb] - indole (1: 1).

A solution of 0.25 g (0.68 mmol) of acid is heated

7-chloro-5-methyl-4-oxo-3-phenyl-3, 5-dihydro-4H-pyridazino_ [4, 5-jb] indole-1-acetic and 0.17 g (1.02 mmol) of 1, 1 '-carbonylbis-1Jî-imidazole in 50 ml of tetrahydrofuran at reflux for 2 h. The mixture is cooled to 0 ° C, 0.19 g (1.02 mmol) of 1,1-dimethylethyl piperazine-1-carboxylate is added and the mixture is stirred at room temperature for 12 h. It is concentrated under reduced pressure, 80 ml of water and 80 ml of dichloromethane are added, the aqueous phase is separated and extract with dichloromethane, the combined organic phases are washed with water and then with a saturated aqueous solution of sodium chloride, they are dried over sodium sulfate, they are filtered, the filtrate is concentrated under reduced pressure and purifies the residue by chromatography on a column of silica gel, eluting with a 9/1 to 6/4 mixture of cyclohexane and ethyl acetate. 0.36 g of compound is isolated and dissolved in 20 ml of dichloromethane, 3 ml of trifluoroacetic acid are added at 0 ° C. and the mixture is stirred at room temperature for 1 h.

It is concentrated under reduced pressure, water and potassium carbonate are added until a basic pH is obtained, 50 ml of dichloromethane are added, the organic phase is decanted, the aqueous phase is extracted with dichloromethane , the organic phases are combined and washed with water and then with a saturated aqueous solution of sodium chloride, they are dried over sodium sulfate, they are filtered, the filtrate is concentrated under reduced pressure and the residue by chromatography on a silica gel column, eluting with a 95/5 mixture of dichloromethane and methanol. 0.28 g of a white solid is isolated.

The hydrochloride is prepared using these 0.28 g (0.64 mmol) of base, dissolved in a mixture of propan-2-ol and methanol, and 10 ml of a hydrochloric acid solution (about 0 , 1N) in propan-2-ol, the product is recrystallized from propan-2-ol, it is collected by filtration, it is rinsed with diethyl ether and it is dried under reduced pressure. 0.145 g of hydrochloride is isolated in the form of a white solid. Melting point: 301-303 ° C.

The following table illustrates the chemical structures and the physical properties of some compounds which can be used according to the invention. Board

Legend

In the column "R ₂ ", cC ₃ H ₅ denotes a cyclopropyl group, cC ₆ H _{1: L} denotes a cyclohexyl group, C ₆ H ₅ denotes a phenyl group and 2-C ₅ H ₄ N denotes a pyridin-2 group -yle.

In the "Salt" column, "-" denotes a compound in the basic state and "HCl" denotes a hydrochloride; the acid: base molar ratio is indicated opposite. The protocols and the results of the tests which have been carried out are described below.

Study of the [ ³ H] Ro5-4864 binding to peripheral benzodiazepine receptors.

The affinity of the compounds of the invention for the peripheral benzodiazepine type receptors (p site, or PBR) was determined.

The p-site receptors can be selectively labeled in rat kidney membranes incubated in the presence of [ ³ H] Ro5-4864. The compounds have been the subject of an in vitro study as to their affinity for these receptors. The animals used are male Sprague Dawley rats (Iffa Credo) from 180 to 300 mg. After decapitation, the kidney is removed and the tissue is homogenized at 4 ° C using a Polytron ™ homogenizer for 2 min at 6/10 of maximum speed in 35 volumes of 50 mM Na ₂ HP0 ₄ phosphate buffer at pH adjusted to 7.5 with NaH ₂ P0 ₄ . The membrane homogenate is filtered through gauze and diluted 10 times with buffer. The [ ³ H] Ro5-4864 (Specific activity: 70-90 Ci / mmol; New England Nuclear), at a concentration of 0.5 nM, is incubated in the presence of 100 μl of the membrane homogenate in a final volume of 1 ml of buffer containing the compound to be tested. After a 3 h incubation at 0 ° C, the membranes are recovered by filtration on Whatman GF / B ™ filters which are washed with twice 4.5 ml of cold incubation buffer (0 ° C). The amount of radioactivity retained by the filter is measured by liquid scintigraphy.

For each concentration of compound studied, the percentage of inhibition of the binding of [ ³ H] Ro5-4864 is determined, then the concentration IC ₅₀ , a concentration which inhibits 50% of the specific binding. The IC ₅₀ values of the most active compounds range from 5 nM to 20 nM.

The compounds which can be used according to the invention are therefore ligands with high affinity for receptors of the peripheral benzodiazepine type. Study of neurotrophic activity.

Test of regeneration of the injured facial nerve by measuring the functional recovery of the palpebral reflex, according to a modification of the method of K. Kujawa et al., Experimental Neurology (1989) 105 80-85.

The lesion of the facial nerve by local freezing leads to degeneration of the distal part of the facial nerve and a loss of the blinking function of the eyelid.

The products to be studied are administered intraperitoneally or orally 2 times a day with a delay of 6 to 8 hours, every day for 10 days (duration of the experiment). The first treatment is administered 30 minutes before the injury. Observation of the animals: the recovery of the function of the eyelids in the injured animals is observed every day, once in the morning from D0 to D5 and 2 times (morning and evening with 6 to 8 h offset) from D6 to D10, before each treatment, according to a theoretical score ranging from 0 to 4. Score 0: open eye, score 1: closed eye with a degree less than half of the eye; score 2: degree of closure between 1/2 and 3/4; score 3: degree of closure greater than 3/4; score 4: eye completely closed. The results are expressed by the report of the AUC ("area under the curve") of the treated group and of the control group. The AUC ratios of the most active compounds are between 1.12 and 1.20. These compounds therefore increase by 12 to 20% the recovery of the palpebral reflex after lesion of the facial nerve.

Motoneuron survival test after section of the facial nerve in 4-day-old rats.

After damage to the facial nerve in the immature rat, the motor neurons in the facial nucleus undergo neuronal death by apoptosis. Neuronal survival is assessed using histological methods and neural counting. 4-day immature rats are anesthetized with pentobarbital (3 mg / kg ip). The right facial nerve is cleared and severed, as it leaves the stylomastoid foramen. After awakening, the pups are returned to their mother and treated, for 7 days, with one or two daily administrations, orally or intraperitoneally, at doses ranging from 1 to 10 mg / kg. 7 days after the lesion, the animals are decapitated, and the brains frozen in isopentane at -40 ° C. The facial nucleus is cut by the cryostat, in sections of 10 μm, in its entirety. The motor neurons are stained with cresyl violet and counted using Histo ™ software (Biocom ™). In this model, the most active compounds increase neuronal survival by around 10 to 30%. The results of the tests described above show that the compounds of general formula (I) promote nerve regeneration.

The compounds of general formula (I) can therefore be used for the preparation of medicaments intended for the prevention and treatment of peripheral neuropathies of different types, such as traumatic or ischemic neuropathies, infectious, alcoholic, medicinal or genetic neuropathies, as well as motor neuron conditions, such as spinal muscular atrophies and amyotrophic lateral sclerosis. These drugs will also find application in the treatment of neurodegenerative diseases of the central nervous system, either of acute type such as cerebrovascular accidents and head and spinal injuries, or of chronic type such as autoimmune diseases (multiple sclerosis), 'Alzheimer's, Parkinson's disease and any other disease in which the administration of neurotrophic factors is supposed to have a therapeutic effect.

The compounds which can be used according to the invention can also be used in the treatment of acute or chronic renal failure, glomerulonephritis, diabetic nephropathy, ischemia and cardiac insufficiency, myocardial infarction, limb ischemia lower, coronary vasospasm, angina pectoris, pathologies associated with heart valves, inflammatory heart disease, side effects due to cardiotoxic drugs or following cardiac surgery, atherosclerosis and its complications thromboembolic, restenosis, graft rejection, conditions related to improper proliferation or migration of smooth muscle cells.

Furthermore, recent data from the literature indicate that the peripheral benzodiazepine receptor could play a fundamental role in the regulation of cell proliferation and cancerization processes. In general, and in comparison with normal tissues, an increased density of peripheral type receptors for benzodiazepines is observed in different types of tumors and cancers.

In human astocytomas, the level of expression of the peripheral benzodiazepine receptor is correlated with the degree of tumor malignancy, the proliferation index and patient survival. In human brain tumors, the increase in the number of peripheral benzodiazepine receptors is used as a diagnostic indication in medical imaging and as a therapeutic target for conjugates formed by a benzodiazepine peripheral receptor ligand and a cytostatic drug. A high density of peripheral benzodiazepine receptors is also observed in ovarian carcinomas and breast cancers. With regard to the latter, it has been demonstrated that the level of expression of peripheral type receptors to benzodiazepines is linked to the aggressive potential of the tumor; moreover, the presence of a peripheral benzodiazepine receptor agonist stimulates the growth of a breast cancer line.

All of these results, which suggests a deleterious function of the peripheral type receptor to Benzodiazepines in the processes of cancerization, constitutes a relevant basis for the search for synthetic ligands specific for the peripheral type receptor to benzodiazepines capable of blocking their effects.

The compounds can therefore be used for the treatment of tumors and cancers.

The compounds of general formula (I) can be used as anti-inflammatories.

Peripheral benzodiazepine receptors are also present in the skin and, as such, the compounds which can be used according to the invention can be used for the prophylaxis or the treatment of cutaneous stresses.

By skin stress is meant the various situations which could cause damage in particular at the level of the epidermis, whatever the agent which causes this stress. This agent can be internal and / or external to the organism, such as a chemical or radical agent, or else external, such as ultraviolet radiation.

Thus, the compounds which can be used according to the invention are intended to prevent and combat skin diseases, such as skin irritations, scabs, erythemas, dysesthetic sensations, heating sensations, pruritus of the skin and / or mucous membranes, aging and can also be used in skin disorders such as, for example, psoriasis, pruritic diseases, herpes, photodermatoses, atopic dermatitis, contact dermatitis, lichens, prurigos , pruritus, insect bites, in fibrosis and other disorders of the maturation of collagens, in immunological disorders or in dermatological conditions such as eczema.

Thus the subject of the present invention is the use of the compounds of general formula (I) for the preparation of pharmaceutical compositions containing an effective dose of at least one compound of general formula (I), in the basic state or of pharmaceutically acceptable salt or solvate, and in admixture, if necessary, with suitable excipients. Said excipients are chosen according to the pharmaceutical form and the desired mode of administration. The pharmaceutical compositions according to the invention can thus be intended for oral, sublingual, subcutaneous, intramuscular, intravenous, topical, intratracheal, intranasal, transdermal, rectal, intraoccular administration. The unit forms of administration can be, for example, tablets, capsules, granules, powders, oral or injectable solutions or suspensions, transdermal patches ("patch"), suppositories. For topical administration, ointments, lotions and eye drops can be considered.

Said unit forms are dosed to allow daily administration of 0.001 to 20 mg of active principle per kg of body weight, depending on the dosage form.

To prepare tablets, a pharmaceutical carrier can be added to the active principle, micronized or not, which can be composed of diluents, such as, for example, lactose, microcrystalline cellulose, starch, and formulation adjuvants such as binders, (polyvinylpyrrolidone, hydroxypropylmethylcellulose, etc.), flow agents such as silica, lubricants such as magnesium stearate, stearic acid, glycerol tribehenate, sodium stearyl fumarate. Wetting agents or surfactants such as sodium lauryl sulfate can also be added.

The production techniques can be direct compression, dry granulation, wet granulation or hot melting. The tablets can be plain, coated, for example with sucrose, or coated with various polymers or other suitable materials. They can be designed to allow rapid, delayed or prolonged release of the active ingredient using polymer matrices or specific polymers used in the coating. To prepare capsules, the active principle is mixed with dry pharmaceutical vehicles (simple mixing, dry or wet granulation, or hot fusion), liquids or semi-solids. The capsules can be hard or soft, film-coated or not, so as to have rapid, prolonged or delayed activity (for example for an enteric form).

A composition in the form of a syrup or elixir or for administration in the form of drops may contain the active principle together with a sweetener, preferably calorie-free, methylparaben or propylparaben as an antiseptic, a flavoring agent and a coloring agent.

Water dispersible powders and granules may contain the active ingredient in admixture with dispersing agents or wetting agents, or dispersing agents such as polyvinylpyrrolidone, as well as with sweeteners and flavor correcting agents.

Suppositories prepared with binders that melt at the rectal temperature, for example cocoa butter or polyethylene glycols, are used for rectal administration.

For parenteral administration, aqueous suspensions, isotonic saline solutions or sterile injectable solutions containing pharmacologically compatible dispersing agents and / or wetting agents, for example propylene glycol or butylene glycol, are used.

The active principle can also be formulated in the form of microcapsules, optionally with one or more carriers or additives, or else with a polymer matrix or with a cyclodextrin (transdermal patches, sustained-release forms).

The topical compositions according to the invention comprise a medium compatible with the skin. They can present themselves especially in the form of aqueous, alcoholic or hydroalcoholic solutions, gels, water-in-oil or oil-in-water emulsions having the appearance of a cream or gel, microemulsions, aerosols, or also in the form of vesicular dispersions containing ionic and / or nonionic lipids. These dosage forms are prepared according to the usual methods of the fields considered.

Finally, the pharmaceutical compositions according to the invention may contain, alongside a compound of general formula (I), other active ingredients which may be useful in the treatment of the disorders and diseases indicated above.

Claims

claims

1. Use of a compound, in the basic state or of an addition salt with an acid, corresponding to the general formula (I)

in which X represents a halogen atom,

Y represents one or more atoms or groups chosen from hydrogen, halogens and hydroxy, methyl and methoxy groups,

R- _L represents a hydrogen atom or a (C ₁ -C ₄ ) alkyl group, R ₂ represents a hydrogen atom, a group (Ci-C linear or branched alkyl, a hydroxy group (C _x -C ₄₎ ) alkyl, a (C ₃ -C ₇ ) cycloalkyl group, a (C ₃ -C ₇ ) cycloalkyl (C ^ Cg) - alkyl group, a phenyl group, a pyridinyl group or a phenyl group (C _x -C ₄ ) alkyl, for the preparation of a medicament intended for the prevention or treatment of a disease linked to the dysfunction of receptors of the peripheral benzodiazepine type.

2. Use according to one of claims 1 and 2, characterized in that the compound of general formula (I) is 7-chloro-5-methyl-l- [2- (4-methylpiperazin-l-yl) - 2-oxoethyl] -4-oxo-3-phenyl-3, 5-dihydro-4fl-pyridazino [4, 5-Jb] - indole or one of its addition salts with an acid.

3. Use according to one of claims 1 and 2, characterized in that the disease is a peripheral neuropathy.

4. Use according to one of claims 1 and 2, characterized in that the disease is renal failure.

5. Use according to one of claims 1 and 2, characterized in that the disease is heart failure.

6. Use according to one of claims 1 and 2, characterized in that the disease is a tumor or cancer.

7. Use according to one of claims 1 and 2, characterized in that the disease is a skin disease.

8. Use according to one of claims 1 and 2, characterized in that the disease is an inflammatory disease.