WO2009044006A2 - 5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxylic acid the pharmaceutically-acceptable esters and salts and solvates thereof for use as medicament - Google Patents

5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxylic acid the pharmaceutically-acceptable esters and salts and solvates thereof for use as medicament Download PDF

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WO2009044006A2
WO2009044006A2 PCT/FR2008/001095 FR2008001095W WO2009044006A2 WO 2009044006 A2 WO2009044006 A2 WO 2009044006A2 FR 2008001095 W FR2008001095 W FR 2008001095W WO 2009044006 A2 WO2009044006 A2 WO 2009044006A2
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inflammation
methylpyrazole
dichlorophenyl
chlorophenyl
rats
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PCT/FR2008/001095
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French (fr)
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WO2009044006A3 (en
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Tiziano Croci
Michel Finet
Fabio Guagnini
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Sanofi-Aventis
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/415—1,2-Diazoles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/08—Drugs for disorders of the urinary system of the prostate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/02—Drugs for disorders of the nervous system for peripheral neuropathies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present invention relates to the use of 5- (4-chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its esters, its pharmaceutically acceptable salts and their solvates for the preparation of medicaments useful for the treatment of pain, neuropathies, inflammation and benign prostatic hypertrophy.
  • Compound A 5- (4-Chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, hereinafter referred to as "Compound A” is described in EP 656 354. It has now been found that this compound has pharmacological activity in different therapeutic areas and that it can be used as a medicament for the treatment of various conditions, in particular for the treatment of pain, neuropathy, inflammation and benign hypertrophy of the skin. prostate.
  • compound A is useful in the treatment of chronic pain, neuropathic pain, neuropathies including diabetic neuropathy, inflammation, including bronchopulmonary asthmatic inflammation or bronchopulmonary allergic inflammation, irritable bowel syndrome, Crohn's disease, inflammation of the intestines.
  • CCI chronic constriction lesion
  • the compound has one of its esters or a pharmaceutically acceptable salt or a solvate thereof may be administered as an active ingredient in the form of pharmaceutical compositions comprising a pharmaceutically acceptable carrier.
  • Compound A, its esters, its pharmaceutically acceptable salts or their solvates may be used at daily doses of 0.01 to 100 mg per kilogram of body weight of the mammal to be treated, preferably at daily doses of 0.1 to 50 mg / kg. In humans, the dose may preferably vary from 0.5 to 4000 mg per day, more particularly from 2.5 to 1000 mg depending on the age of the subject to be treated or the type of treatment: prophylactic or curative.
  • the active ingredient can be administered in unit dosage form, in admixture with conventional pharmaceutical carriers, animals and humans.
  • suitable unit dosage forms include oral forms, such as tablets, capsules, powders, granules and oral solutions or suspensions, sublingual and oral forms of administration, aerosols, implants, forms of subcutaneous, intramuscular, intravenous, intranasal or intraocular administration and forms of rectal administration.
  • the active principle is generally in dosage units containing from 0.5 to 1000 mg, advantageously from 1 to 500 mg, preferably from 2 to 200 mg of said active ingredient per dosage unit for daily administrations.
  • a wetting agent such as sodium lauryl sulphate may be added to the active ingredient, whether it is micronized or not, and the mixture is mixed with a pharmaceutical vehicle such as silica, starch, lactose, magnesium stearate, talc, mannitol, croscaramellose sodium, corn starch, hydroxypropyl methylcellulose or the like.
  • a pharmaceutical vehicle such as silica, starch, lactose, magnesium stearate, talc, mannitol, croscaramellose sodium, corn starch, hydroxypropyl methylcellulose or the like.
  • the sucrose tablets, various polymers or other suitable materials may be coated or treated in such a way that they have prolonged or delayed activity and continuously release a predetermined amount of active ingredient.
  • a unitary form of administration of a compound according to the invention in tablet form may comprise the following components:
  • a preparation in capsules is obtained by mixing the active ingredient with a diluent such as a glycol or a glycerol ester and incorporating the mixture obtained into soft or hard gelatin capsules.
  • a preparation in syrup or elixir form may contain the active ingredient together with a sweetener, preferably calorie-free, methylparaben and propylparaben as antiseptic, as well as a flavoring agent and a suitable colorant.
  • the water-dispersible powders or granules may contain the active ingredient in admixture with dispersing agents, wetting agents, or suspending agents, such as polyvinylpyrrolidone, as well as with sweeteners or scavengers. taste.
  • suppositories are used which are prepared with binders melting at the rectal temperature, for example, cocoa butter or polyethylene glycols.
  • aqueous suspensions, isotonic solutions or sterile and injectable solutions which contain dispersing agents and / or pharmaceutically compatible solubilizing agents, for example propylene glycol or polyethylene glycol, are used.
  • a cosolvent can be used: an alcohol such as ethanol, a glycol such as polyethylene glycol or propylene glycol and a hydrophilic surfactant such as
  • the active principle can be solubilized by a triglyceride or a glycerol ester.
  • patches in multilaminated form or reservoirs in which the active ingredient is in alcoholic solution can be used.
  • the active ingredient may also be formulated in the form of microcapsules or microspheres, optionally with one or more supports or additives.
  • the sustained-release forms useful in the case of chronic treatment the implants can be used. These can be prepared as an oily suspension or as a suspension of microspheres in an isotonic medium.
  • TEST 1 Effect of compound A on cell migration in the bronchoalveolar space after stimulation with ovalbumin.
  • Balb / c mice weighing between 28 and 30 g were sensitized on day 1 and day 11 by intraperitoneal injection of a 0.05% solution of ovalbumin (in a 3% AI gel ( OH) 3 ) in a volume of 0.2 ml / animal.
  • mice were stimulated by intratracheal exposure of 10 ⁇ l of a 1% solution of ovalbumin (dissolved in sterile isotonic water). 24 hours after ovalbumin injection, the animals were anesthetized with pentobarbital and bronchoalveolar lavage was performed. The washing fluids were recovered, centrifuged and the cells resuspended. The number of cells was counted by differentiating, according to standard morphological criteria, eosinophilic, neutrophilic and mononuclear cells.
  • Intratracheal stimulation with ovalbumin induced a significant increase in the number of mononuclear and eosinophilic cells in the bronchoalveolar space of the mice: compared to the control group, respectively from 26800 to 47700 and from 480 to 11500 cells.
  • the effect of compound A treatment on the ovalbumin-induced recruitment of these cells was studied.
  • Compound A or vehicle water / Tween 80 was administered to animals 1 hour prior to ovalbumin stimulation at doses ranging from 0.1 to 10 mg / kg / i.p. in a volume of 0.1 ml / 10 g of weight. The results obtained are collated in Table 1 below.
  • total number was 0.9 mg / kg.
  • compound A In intraperitoneal administration one hour prior to stimulation with an allergenic agent, compound A thus inhibits the migration of inflammatory cells in a dose-dependent manner 24 hours later, the mononuclear cells being more significantly inhibited than eosinophils. Mononuclear cells have not been characterized. Conventionally, the mononuclear cells in the bronchoalveolar fluid of the control mice are predominantly macrophagic while the cells that migrate after stimulation are presumably predominantly lymphocyte but have not been differentiated by flow cytofluorometry.
  • TEST 2 Effect of compound A on the nociceptive response in neuropathic rats.
  • rats with CCI The sciatic nerve of each rat was isolated and, near the sciatic nerve trifurcation, four loose ligatures were made around the nerve.
  • the rats thus treated are hereinafter referred to as "rats with CCI”.
  • Compound A was administered orally and daily for 8 days at the dose of 30 mg / kg from the 7th day after surgery (ligated) by the procedure below:
  • Compound A was dissolved in the vehicle consisting of distilled water (pH 7.5) containing 5% DMSO (dimethylsulfoxide) and 5% Tween 80 and administered by gavage at a rate of 2 ml / kg of body weight.
  • vehicle consisting of distilled water (pH 7.5) containing 5% DMSO (dimethylsulfoxide) and 5% Tween 80 and administered by gavage at a rate of 2 ml / kg of body weight.
  • LTiyperalgésie thermal and mechanical allodynia response were recorded before surgery, then the 7th day and the 14th day (24 hours after the 7 th administration) after surgery and finally the 14th day ( 2 hours after the last administration of the compound A).
  • Thermal hyperalgesia was evaluated by recording latencies of paw retraction at radiation thermal stimulation according to the protocol or plantar test described by Hargreaves et al. Pain, 32 (1988) 77-88 using the analgesia meter marketed by the Italian company Ugo Basile, Varese, Italy.
  • Table 2 Effect of compound A on thermal hyperalgesia in rats with CCI and control rats.
  • the results are mean ⁇ SD for 5 to 8 rats.
  • the results are mean ⁇ EET for 5 to 8 rats.
  • TEST 3 Effect of compound A on neurite elongation
  • the posterior L4, L5 and L6 dorsal spinal ganglia with attached roots (DRG) were removed by micro-dissection in na ⁇ ve rats (controls without CCI) and neuropathic rats (rats with CCI) according to the protocol described in Example 2 above after anesthesia with pentobarbital sodium (100 mg / kg body weight ip). In both cases, the rats were adult Charles River CrI: CD BR rats weighing 400-500 g.
  • lymph nodes removed were incubated in medium of EEAGLE modified with DULBECCO medium (DMEM medium) containing 0.5 mg / ml of trypsin, 1 mg / ml of collagenase and 0.1 mg / ml of DNAase for 90 minutes. 37 ° C. A soy trypsin inhibitor was then added to neutralize the trypsin and the resulting suspension was centrifuged at 1000 g for one minute.
  • DMEM medium DULBECCO medium
  • the neurons contained in the centrifuge cake were suspended in DMEM medium containing 10% calf serum, 10% horse serum, 2 mM glutamine, 0.1 mg / ml penicillin / streptomycin and 2.5 mg / ml DNase and distributed in Petri dishes (diameter 35 mm) coated with Matrigel. The neurons were then incubated at 37 ° C in the presence of 95% CO 2 and 5% O 2 in a basal medium under different experimental conditions:
  • NGF nerve growth factor
  • next day cultures were washed and incubated with a second universal peroxidase-conjugated antibody (according to the ABC technique using an avidin-biotin enzyme complex with the kit sold under the name Vectastain® by VECTOR Laboratories in Burlingame, California. , USA) and stained with a solution of 3,3'-diaminobenzidine (DAB).
  • DAB 3,3'-diaminobenzidine
  • DRG explants were measured on each photograph in a random manner using SCION Image NIH software (National Institute of Health Bethesda, Maryland USA).
  • the exogenous NGF leads to an increase in elongation DRG neurites which is the maximum day 3 after injury. The 7 th day, the exogenous NGF is unable to cause further increase in neurite elongation DRG, probably due to activation of Schwann cells (regeneration process).
  • Benign prostatic hypertrophy (BPH) was induced in 10 male CD rats by administration of 18.75 mg testosterone per day subcutaneously for 24 days. Compound A was then administered orally in combination with
  • TEST 5 Isometric measurement The following ex vivo test was performed on the ventral lobe of the prostate of rats in which benign hypertrophy was induced according to the procedure described in test 4.
  • the prostate was rapidly excised in male rats Crl: CD ® Charles River) weighing 300-35Og and placed in Krebs bicarbonate buffer (118.4 mM NaCl, 4.7 mM KCl, CaCl 2 2, 5mM, 1.2mM KH 2 PO 4 , 25mM NaHCO 3 , 11.7mM glucose).
  • the prostate was cleared of surrounding tissue and the ventral portion was isolated and cut into two lobes. Each lobe was mounted longitudinally in a 20 ml isolated organ chamber for recording the isometric tension, put in Krebs bicarbonate buffer at 37 ° C, oxygenated with 95% O 2 - 5% CO 2 and stretched to a passive voltage of 250 mg. After an equilibration period during which the lobe was washed with Krebs buffer as defined above 4 times for 10 minutes, the norepinephrine response curve (1 nM - 0.1 mM) was obtained in the presence 0.5 ⁇ M of DMI (desmethylimipramine, Ciba) and 30 ⁇ M of hydrocortisone hemisuccinate (Flebocortid®, Lepetit).
  • DMI desmethylimipramine
  • Ciba hydrocortisone hemisuccinate
  • the isometric force changes were recorded using the PowerLab data acquisition system (AD Instruments Pty Ltd, Castle HiII, Australia).
  • the generation of isometric force was measured as the percentage of contraction induced by 40 mM KCI (100% maximal tissue contraction) added at the end of the experiment after the highest concentration of norepinephrine.
  • Compound A administered at 10 or 30 mg / kg body weight showed a normal contraction capacity to norepinephrine. Compound A thus restores the contractibility of the tested prostate sample.
  • the examples show that the compound A, its esters, its pharmaceutically acceptable salts and / or their solvates are useful for the treatment or for the preparation of a medicament intended for the treatment of chronic pain, neuropathic pain, neuropathies, especially neuropathies.
  • Diabetic neuropathy, inflammation, including bronchopulmonary asthmatic inflammation or bronchopulmonary allergic inflammation, irritable bowel syndrome, Crohn's disease, inflammation of the intestines and may be used for the preparation of a drug intended for such treatment.
  • the present invention also relates to a method of treatment of the pathologies indicated above which comprises the administration to a patient of an effective dose of the compound A its esters, its pharmaceutically acceptable salts and / or or their solvates.

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Abstract

The invention relates to 5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxylic acid the pharmaceutically-acceptable esters and salts and solvates thereof for use as medicament. Application : Treatment of pain, neuropathies, inflammation and benign hypertrophy of the prostate.

Description

L'acide 5-(4-chlorophényl)-l-(2,4-dichlorophényl)-4-méthylpyrazole-3- carboxylique, ses esters, ses sels pharmaceutiquement acceptables et leurs solvates pour leur utilisation comme médicament. 5- (4-Chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its esters, pharmaceutically acceptable salts and solvates for use as a medicament.
La présente invention a pour objet l'utilisation de l'acide 5-(4-chlorophényl)- l-(2,4-dichlorophényl)-4-méthylpyrazole-3-carboxylique, de ses esters, de ses sels pharmaceutiquement acceptables et de leurs solvates pour la préparation de médicaments utiles pour le traitement de la douleur, des neuropathies, de l'inflammation et de l'hypertrophie bénigne de la prostate.The present invention relates to the use of 5- (4-chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its esters, its pharmaceutically acceptable salts and their solvates for the preparation of medicaments useful for the treatment of pain, neuropathies, inflammation and benign prostatic hypertrophy.
L'acide 5-(4-chlorophényl)-l-(2,4-dichlorophényl)-4-méthylpyrazole-3- carboxylique, ci-après dénommé « composé A » est décrit dans le brevet EP 656 354. On a maintenant trouvé que ce composé présente une activité pharmacologique dans différents domaines thérapeutiques et qu'il peut être utilisé comme médicament pour le traitement de différentes affections, notamment pour le traitement de la douleur, des neuropathies, de l'inflammation et de l'hypertrophie bénigne de la prostate.5- (4-Chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, hereinafter referred to as "Compound A" is described in EP 656 354. It has now been found that this compound has pharmacological activity in different therapeutic areas and that it can be used as a medicament for the treatment of various conditions, in particular for the treatment of pain, neuropathy, inflammation and benign hypertrophy of the skin. prostate.
Plus particulièrement, selon la présente invention, le composé A, ses esters, ses sels pharmaceutiquement acceptables et/ou leurs solvates sont utiles dans le traitement des douleurs chroniques, des douleurs neuropathiques, des neuropathies notamment de la neuropathie diabétique, de l'inflammation, notamment de l'inflammation asthmatique broncho-pulmonaire ou inflammation allergique broncho-pulmonaire, du syndrome du colon irritable, de la maladie de Crohn, de l'inflammation des intestins.More particularly, according to the present invention, compound A, its esters, its pharmaceutically acceptable salts and / or their solvates are useful in the treatment of chronic pain, neuropathic pain, neuropathies including diabetic neuropathy, inflammation, including bronchopulmonary asthmatic inflammation or bronchopulmonary allergic inflammation, irritable bowel syndrome, Crohn's disease, inflammation of the intestines.
Les effets du composé A ont été étudiés chez des animaux dans les modèles expérimentaux ci-après. A - chez les souris sur un modèle d'asthme allergique induit par ovalbumine selon le protocole de Kips etal., (EurRespirJ. 2003, 22, 374-382). La sensibilisation par injection intrapéritonéale d'un allergène, l'ovalbumine, en présence d'alun comme adjuvant recrute et active des sous-populations de lymphocytes T, les CD4+ et certains Th2, qui activent les lymphocytes B à produire des immunoglobulines IgE spécifiques de l'ovalbumine. Ces cellules jouent également un rôle de mémoire. Lors de la seconde exposition ou stimulation, l'ovalbumine active les mastocytes par liaison aux IgE spécifiques et réactivent les lymphocytes T en particulier les CD4+ et les Th2. Il se produit une infiltration des cellules lymphocytes T et des éosinophiles dans les tissus broncho-pulmonaires suivie d'une libération de médiateurs qui entraînent des lésions tissulaires. Cette infiltration des cellules inflammatoires est la conséquence de l'activation des cellules mononucléaires (mastocytes et macrophages qui constituent la première barrière de défense au niveau de l'épithélium bronchique et lymphocytes T) stimulées par activation antigénique lors de la stimulation par l'ovalbumine. Ce type de modèle allergique chez la souris a permis de comprendre les rôles clés joués par les lymphocytes T CD4+ et par les Th2 et les cytokines libérées par ces cellules (Kυmar & Foster, Am J Resp CeII Mol Biol, 27, 267-272, 2002). B - chez des rats neuropathiques chez lesquels une lésion chronique par constriction (dénommée ci-après CCI) a été induite selon le protocole décrit par Bennet étal. Pain, 33 (1988) 87-107.The effects of compound A have been studied in animals in the experimental models below. A - in mice on a model of allergic asthma induced by ovalbumin according to the protocol of Kips et al., (EurRespirJ 2003, 22, 374-382). Sensitization by intraperitoneal injection of an allergen, ovalbumin, in the presence of alum as an adjuvant recruits and activates subpopulations of T lymphocytes, CD4 + and some Th2, which activate B cells to produce IgE immunoglobulins specific for ovalbumin. These cells also play a memory role. In the second exposure or stimulation, ovalbumin activates mast cells by binding to specific IgE and reactivates T lymphocytes, particularly CD4 + and Th2. There is infiltration of T cells and eosinophils in bronchopulmonary tissues followed by release of mediators that result in tissue damage. This infiltration of inflammatory cells is the consequence of the activation of mononuclear cells (mast cells and macrophages which constitute the first defense barrier at the level of the bronchial epithelium and T lymphocytes) stimulated by antigenic activation during stimulation by ovalbumin. This type of allergic model in the mouse has made it possible to understand the key roles played by the CD4 + T lymphocytes and by the Th2 and cytokines released by these cells (Kümar & Foster, Am J Resp CeII Mol Biol, 27, 267-272, 2002). B - in neuropathic rats in which a chronic constriction injury (hereinafter referred to as CCI) was induced according to the protocol described by Bennet et al. Bread, 33 (1988) 87-107.
C - sur les ganglions dorsaux rachidiens avec racines de rats neuropathiques chez lesquels une lésion chronique par constriction (dénommée ci-après CCI) a été induite selon le protocole indiqué ci-dessus, en utilisant le protocole de C. Sommer et al. Journal of Neuropathology and Expérimental Neurology et R. Lindsay et al. Development Biology, Vol. 112, 30-48 (1985).C - on the spinal dorsal ganglia with roots of neuropathic rats in which a chronic constriction lesion (hereinafter referred to as CCI) was induced according to the protocol indicated above, using the protocol of C. Sommer et al. Journal of Neuropathology and Experimental Neurology and R. Lindsay et al. Development Biology, Vol. 112, 30-48 (1985).
D - chez des rats chez lesquels une hypertrophie bénigne de la prostate a été induite par la testostérone ( Van Copenolle F. et al., Am. J. Physiol. Endocrinol. Metab. 280:E120-E129, 2001; Fujimoto N. et al., Cancer Sci: 95: 711-715, 2004).D - in rats in which a benign prostatic hypertrophy was induced by testosterone (Van Copenolle F. et al., Am J Physiol Endocrinol, Metab 280: E120-E129, 2001; Fujimoto N. et al. al., Cancer Sci: 95: 711-715, 2004).
Le composé A un de ses esters ou un de ses sels pharmaceutiquement acceptables ou un de leurs solvates peut être administré, en tant que principe actif, sous forme de compositions pharmaceutiques comprenant un véhicule pharmaceutiquement acceptable. Le composé A, ses esters, ses sels pharmaceutiquement acceptables ou leurs solvates peuvent être utilisés à des doses journalières de 0,01 à 100 mg par kilo de poids corporel du mammifère à traiter, de préférence à des doses journalières de 0,1 à 50 mg/kg. Chez l'être humain, la dose peut varier de préférence de 0,5 à 4000 mg par jour, plus particulièrement de 2,5 à 1000 mg selon l'âge du sujet à traiter ou le type de traitement : prophylactique ou curatif.The compound has one of its esters or a pharmaceutically acceptable salt or a solvate thereof may be administered as an active ingredient in the form of pharmaceutical compositions comprising a pharmaceutically acceptable carrier. Compound A, its esters, its pharmaceutically acceptable salts or their solvates may be used at daily doses of 0.01 to 100 mg per kilogram of body weight of the mammal to be treated, preferably at daily doses of 0.1 to 50 mg / kg. In humans, the dose may preferably vary from 0.5 to 4000 mg per day, more particularly from 2.5 to 1000 mg depending on the age of the subject to be treated or the type of treatment: prophylactic or curative.
Dans les compositions pharmaceutiques de la présente invention pour l'administration orale, sublinguale, sous-cutanée, intramusculaire, intraveineuse, transdermique, locale ou rectale, le principe actif peut être administré sous forme unitaire d'administration, en mélange avec des supports pharmaceutiques classiques, aux animaux et aux êtres humains. Les formes unitaires d'administration appropriées comprennent les formes par voie orale, telles que les comprimés, les gélules, les poudres, les granules et les solutions ou suspensions orales, les formes d'administration sublinguale et buccale, les aérosols, les implants, les formes d'administration sous-cutanée, intramusculaire, intraveineuse, intranasale ou intraoculaire et les formes d'administration rectale.In the pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous administration, transdermal, local or rectal, the active ingredient can be administered in unit dosage form, in admixture with conventional pharmaceutical carriers, animals and humans. Suitable unit dosage forms include oral forms, such as tablets, capsules, powders, granules and oral solutions or suspensions, sublingual and oral forms of administration, aerosols, implants, forms of subcutaneous, intramuscular, intravenous, intranasal or intraocular administration and forms of rectal administration.
Dans les compositions pharmaceutiques de la présente invention, le principe actif est généralement en unités de dosage contenant de 0,5 à 1000 mg, avantageusement de 1 à 500 mg, de préférence de 2 à 200 mg dudit principe actif par unité de dosage pour les administrations quotidiennes.In the pharmaceutical compositions of the present invention, the active principle is generally in dosage units containing from 0.5 to 1000 mg, advantageously from 1 to 500 mg, preferably from 2 to 200 mg of said active ingredient per dosage unit for daily administrations.
Lorsque l'on prépare une composition solide sous forme de comprimé, on peut ajouter au principe actif, micronisé ou non, un agent mouillant tel que le laurylsulfate de sodium et on mélange le tout avec un véhicule pharmaceutique, tel que la silice, l'amidon, le lactose, le stéarate de magnésium, le talc, mannitol, croscaramellose sodique, amidon de maïs, hydroxypropyl-méthylcellulose ou analogues. On peut enrober les comprimés de saccharose, de divers polymères ou d'autres matières appropriées ou encore les traiter de telle sorte qu'ils aient une activité prolongée ou retardée et qu'ils libèrent d'une façon continue une quantité prédéterminée de principe actif.When a solid composition in the form of a tablet is prepared, a wetting agent such as sodium lauryl sulphate may be added to the active ingredient, whether it is micronized or not, and the mixture is mixed with a pharmaceutical vehicle such as silica, starch, lactose, magnesium stearate, talc, mannitol, croscaramellose sodium, corn starch, hydroxypropyl methylcellulose or the like. The sucrose tablets, various polymers or other suitable materials may be coated or treated in such a way that they have prolonged or delayed activity and continuously release a predetermined amount of active ingredient.
A titre d'exemple, une forme unitaire d'administration d'un composé selon l'invention sous forme de comprimé peut comprendre les composants suivants :By way of example, a unitary form of administration of a compound according to the invention in tablet form may comprise the following components:
Composé A 50,0 mg Mannitol 223,75 mgCompound A 50.0 mg Mannitol 223.75 mg
Croscaramellose sodique 6,0 mgCroscaramellose sodium 6.0 mg
Amidon de maïs 15,0 mgCorn starch 15.0 mg
Hydroxypropyl-méthylcellulose 2,25 mgHydroxypropyl methylcellulose 2.25 mg
Stéarate de magnésium 3,0 mg.Magnesium stearate 3.0 mg.
On obtient une préparation en gélules en mélangeant le principe actif avec un diluant tel qu'un glycol ou un ester de glycérol et en incorporant le mélange obtenu dans des gélules molles ou dures. Une préparation sous forme de sirop ou d'élixir peut contenir le principe actif conjointement avec un édulcorant, acalorique de préférence, du méthylparaben et du propylparaben comme antiseptique, ainsi qu'un agent donnant du goût et un colorant approprié. Les poudres ou les granules dispersibles dans l'eau peuvent contenir le principe actif en mélange avec des agents de dispersion, des agents mouillants, ou des agents de mise en suspension, comme la polyvinylpyrrolidone, de même qu'avec des édulcorants ou des correcteurs du goût.A preparation in capsules is obtained by mixing the active ingredient with a diluent such as a glycol or a glycerol ester and incorporating the mixture obtained into soft or hard gelatin capsules. A preparation in syrup or elixir form may contain the active ingredient together with a sweetener, preferably calorie-free, methylparaben and propylparaben as antiseptic, as well as a flavoring agent and a suitable colorant. The water-dispersible powders or granules may contain the active ingredient in admixture with dispersing agents, wetting agents, or suspending agents, such as polyvinylpyrrolidone, as well as with sweeteners or scavengers. taste.
Pour une administration rectale, on recourt à des suppositoires qui sont préparés avec des liants fondant à la température rectale, par exemple, du beurre de cacao ou des polyéthylèneglycols.For rectal administration, suppositories are used which are prepared with binders melting at the rectal temperature, for example, cocoa butter or polyethylene glycols.
Pour une administration parentérale, on utilise des suspensions aqueuses, des solutions isotoniques ou des solutions stériles et injectables, qui contiennent des agents de dispersion et/ou des agents solubilisants pharmaceutiquement compatibles, par exemple le propylèneglycol ou le polyéthyléneglycol.For parenteral administration, aqueous suspensions, isotonic solutions or sterile and injectable solutions which contain dispersing agents and / or pharmaceutically compatible solubilizing agents, for example propylene glycol or polyethylene glycol, are used.
Ainsi, pour préparer une solution aqueuse injectable par voie intraveineuse, on peut utiliser un cosolvant : un alcool tel que l'éthanol, un glycol tel que le polyéthyléneglycol ou le propylèneglycol et un tensioactif hydrophile tel que leThus, to prepare an injectable aqueous solution intravenously, a cosolvent can be used: an alcohol such as ethanol, a glycol such as polyethylene glycol or propylene glycol and a hydrophilic surfactant such as
Tween® 80. Pour préparer une solution huileuse injectable par voie intramusculaire, on peut solubiliser le principe actif par un triglycéride ou un ester de glycérol.Tween ® 80. To prepare an injectable oily solution by the intramuscular route, the active principle can be solubilized by a triglyceride or a glycerol ester.
Pour l'administration transdermique, on peut utiliser des patches sous forme multilaminée ou réservoirs dans lequel le principe actif est en solution alcoolique.For transdermal administration, patches in multilaminated form or reservoirs in which the active ingredient is in alcoholic solution can be used.
Le principe actif peut être formulé également sous forme de microcapsules ou microsphères, éventuellement avec un ou plusieurs supports ou additifs. Parmi les formes à libération prolongée utiles dans le cas de traitement chroniques, on peut utiliser les implants. Ceux-ci peuvent être préparés sous forme de suspension huileuse ou sous forme de suspension de microsphères dans un milieu isotonique.The active ingredient may also be formulated in the form of microcapsules or microspheres, optionally with one or more supports or additives. Among the sustained-release forms useful in the case of chronic treatment, the implants can be used. These can be prepared as an oily suspension or as a suspension of microspheres in an isotonic medium.
La présente invention va maintenant être décrite plus en détail par les tests expérimentaux ci-après. TEST 1 : Effet du composé A sur le migration des cellules dans l'espace bronchoalvéolaire après stimulation par l'ovalbumine.The present invention will now be described in more detail by the experimental tests hereinafter. TEST 1: Effect of compound A on cell migration in the bronchoalveolar space after stimulation with ovalbumin.
Des souris Balb/c pesant entre 28 et 30 g (Charles River) ont été sensibilisées au jour 1 et au jour 11 par injection intrapéritonéale d'une solution d'ovalbumine 0,05 % (dans un gel à 3 % d' AI(OH)3) dans un volume de 0,2 ml/animal.Balb / c mice weighing between 28 and 30 g (Charles River) were sensitized on day 1 and day 11 by intraperitoneal injection of a 0.05% solution of ovalbumin (in a 3% AI gel ( OH) 3 ) in a volume of 0.2 ml / animal.
Au jour 19, les souris ont été stimulées par une exposition intratrachéale de 10 μl d'une solution à 1 % d'ovalbumine (dissoute dans de l'eau isotonique stérile). 24 heures après l'injection d'ovalbumine, les animaux ont été anesthésiés au pentobarbital et on a réalisé un lavage bronchoalvéolaire. Les fluides de lavage ont été récupérés, centrifugés puis les cellules remises en suspension. On a compté le nombre de cellules en différenciant, d'après les critères morphologiques standard, les cellules éosinophiles, neutrophiles et mononucléaires. La stimulation intratrachéale par l'ovalbumine a induit une augmentation importante du nombre des cellules mononucléaires et éosinophiles dans l'espace bronchoalvéolaire des souris : par rapport au groupe contrôle, respectivement de 26800 à 47700 et de 480 à 11500 cellules. On a étudié l'effet du traitement par le composé A sur le recrutement, induit par l'ovalbumine, de ces cellules. Le composé A ou son véhicule (eau/Tween 80) a été administré aux animaux 1 heure avant la stimulation par l'ovalbumine à des doses variant de 0,1 à 10 mg/kg/i.p. dans un volume de 0,1 ml/10 g de poids. Les résultats obtenus sont rassemblés dans le tableau 1 ci-aprèsAt day 19, the mice were stimulated by intratracheal exposure of 10 μl of a 1% solution of ovalbumin (dissolved in sterile isotonic water). 24 hours after ovalbumin injection, the animals were anesthetized with pentobarbital and bronchoalveolar lavage was performed. The washing fluids were recovered, centrifuged and the cells resuspended. The number of cells was counted by differentiating, according to standard morphological criteria, eosinophilic, neutrophilic and mononuclear cells. Intratracheal stimulation with ovalbumin induced a significant increase in the number of mononuclear and eosinophilic cells in the bronchoalveolar space of the mice: compared to the control group, respectively from 26800 to 47700 and from 480 to 11500 cells. The effect of compound A treatment on the ovalbumin-induced recruitment of these cells was studied. Compound A or vehicle (water / Tween 80) was administered to animals 1 hour prior to ovalbumin stimulation at doses ranging from 0.1 to 10 mg / kg / i.p. in a volume of 0.1 ml / 10 g of weight. The results obtained are collated in Table 1 below.
Tableau 1 : pourcentage d'inhibitionTable 1: percentage inhibition
Figure imgf000006_0001
L'inhibition de la migration cellulaire a été plus importante sur les cellules mononuclées que sur les éosinophiles ; ainsi à 10 mg/kg, l'inhibition de la migration des mononucléaires était totale alors que celle des éosinophiles n'a atteint que 62 %. La dose efficace 50 (DE50) qui inhibe la migration des cellules inflammatoires
Figure imgf000006_0001
Inhibition of cell migration was greater on mononuclear cells than on eosinophils; thus, at 10 mg / kg, the inhibition of mononuclear migration was complete whereas that of eosinophils reached only 62%. The effective dose 50 (DE 50 ) that inhibits the migration of inflammatory cells
(nombre total) était 0,9 mg/kg.(total number) was 0.9 mg / kg.
En administration intrapéritonéale une heure avant la stimulation par un agent allergène, le composé A inhibe donc de façon dose dépendante la migration des cellules inflammatoires mesurée 24 heures plus tard, les cellules mononuclées étant inhibées de manière plus importante que les éosinophiles. Les cellules mononuclées n'ont pas été caractérisées. De manière classique les cellules mononuclées dans le liquide bronchoalvéolaire des souris contrôles sont à prédominance macrophagique tandis que les cellules qui migrent après stimulation sont vraisemblablement à prédominance lymphocytaire, mais n'ont pas été différenciées par cytofluorométrie de flux.In intraperitoneal administration one hour prior to stimulation with an allergenic agent, compound A thus inhibits the migration of inflammatory cells in a dose-dependent manner 24 hours later, the mononuclear cells being more significantly inhibited than eosinophils. Mononuclear cells have not been characterized. Conventionally, the mononuclear cells in the bronchoalveolar fluid of the control mice are predominantly macrophagic while the cells that migrate after stimulation are presumably predominantly lymphocyte but have not been differentiated by flow cytofluorometry.
TEST 2 : Effet du composé A sur la réponse nociceptive chez les rats neuropathiαues.TEST 2: Effect of compound A on the nociceptive response in neuropathic rats.
Le nerf sciatique de chaque rat a été isolé et, à proximité de la trifurcation des nerfs sciatiques, quatre ligatures lâches ont été faites autour du nerf. Les rats ainsi traités sont dénommés ci-après « rats avec CCI ».The sciatic nerve of each rat was isolated and, near the sciatic nerve trifurcation, four loose ligatures were made around the nerve. The rats thus treated are hereinafter referred to as "rats with CCI".
Des rats chez lesquels le nerf sciatique a été isolé, mais sans ligature, ont été utilisés comme animaux témoins (rats témoins).Rats in which the sciatic nerve was isolated, but without ligation, were used as control animals (control rats).
Le composé A a été administré par voie orale quotidiennement et pendant 8 jours à la dose de 30 mg/kg à partir du 7eme jour après l'intervention chirurgicale (ligature) selon le mode opératoire ci-après :Compound A was administered orally and daily for 8 days at the dose of 30 mg / kg from the 7th day after surgery (ligated) by the procedure below:
- le composé A a été dissous dans le véhicule constitué d'eau distillée (pH 7,5) contenant 5% de DMSO (diméthylsulfoxyde) et 5% de Tween 80 et administré par gavage à raison de 2 ml/kg de poids corporel.Compound A was dissolved in the vehicle consisting of distilled water (pH 7.5) containing 5% DMSO (dimethylsulfoxide) and 5% Tween 80 and administered by gavage at a rate of 2 ml / kg of body weight.
LTiyperalgésie thermique et la réponse de l'allodynie mécanique ont été enregistrées avant l'intervention chirurgicale, puis le 7ème jour et le 14 ème jour (24 heures après la 7ème administration) après l'intervention chirurgicale et enfin le 14ème jour (2 heures après la dernière administration du composé A). L/hyperalgésie thermique a été évaluée par enregistrement des latences de retrait des pattes à une stimulation thermique par rayonnement selon le protocole ou test plantaire décrit par Hargreaves et al. Pain, 32 (1988) 77-88 en utilisant l'appareil de mesure de l'analgésie commercialisé par la Société italienne Ugo Basile, Varese, Italie.LTiyperalgésie thermal and mechanical allodynia response were recorded before surgery, then the 7th day and the 14th day (24 hours after the 7 th administration) after surgery and finally the 14th day ( 2 hours after the last administration of the compound A). Thermal hyperalgesia was evaluated by recording latencies of paw retraction at radiation thermal stimulation according to the protocol or plantar test described by Hargreaves et al. Pain, 32 (1988) 77-88 using the analgesia meter marketed by the Italian company Ugo Basile, Varese, Italy.
L'allodynie mécanique a été déterminée selon la méthode décrite par Villeti et al. (The Journal of Pharmacology and Expérimental Therapeutics, 306 :804- 814,2003) en utilisant l'instrument de mesure de la réponse à une stimulation mécanique (Plantar Aesthesio meter) commercialisé par la Société italienne Ugo Basile.Mechanical allodynia was determined according to the method described by Villeti et al. (The Journal of Pharmacology and Experimental Therapeutics, 306: 804-814,2003) using the instrument for measuring the response to mechanical stimulation (Plantar Aesthesio meter) marketed by the Italian company Ugo Basile.
Les résultats obtenus sont rassemblés dans les tableaux 2 et 3 ci-après.The results obtained are collated in Tables 2 and 3 below.
Tableau 2 : Effet du composé A sur l'hyperalgésie thermique chez les rats avec CCI et les rats témoins.Table 2: Effect of compound A on thermal hyperalgesia in rats with CCI and control rats.
Figure imgf000008_0001
Figure imgf000008_0001
Les résultats sont des moyennes ± ET pour 5 à 8 rats.The results are mean ± SD for 5 to 8 rats.
*p<0,05, **p<0,001 par rapport aux rats CCI + Véhicule, oop<0,001 par rapport aux témoins (Anovarep 2 passages plus test Newman-Keuls). Tableau 3 : Effet du composé A sur l'allodynie mécanique chez les rats avec CCI et les rats témoins.* p <0.05, ** p <0.001 compared to CCI + rats Vehicle, oo p <0.001 compared to controls (Anovarep 2 passages plus Newman-Keuls test). Table 3: Effect of compound A on mechanical allodynia in rats with CCI and control rats.
Figure imgf000009_0001
Figure imgf000009_0001
Les résultats sont des moyennes ± EET pour 5 à 8 rats.The results are mean ± EET for 5 to 8 rats.
*p<0,05, **p<0,001 par rapport aux rats CCI + Véhicule, oop<0,001 par rapport aux témoins (Anovarep 2 passages plus test Newman-Keuls).* p <0.05, ** p <0.001 compared to CCI + rats Vehicle, oo p <0.001 compared to controls (Anovarep 2 passages plus Newman-Keuls test).
TEST 3 : Effet du composé A sur l'élongation des neuritesTEST 3: Effect of compound A on neurite elongation
Les ganglions dorsaux rachidiens postérieurs L4, L5 et L6 avec les racines attachées (DRG) ont été prélevés par micro-dissection sur des rats naïfs (témoins sans CCI) et des rats rendus neuropathiques (rats avec CCI) selon le protocole décrit dans l'exemple 2 ci-dessus après anesthésie avec du pentobarbital sodique (100 mg/kg de poids corporel par voie i.p.). Dans les deux cas, les rats étaient des rats adultes Charles River CrI : CD BR pesant 400 à 500 g.The posterior L4, L5 and L6 dorsal spinal ganglia with attached roots (DRG) were removed by micro-dissection in naïve rats (controls without CCI) and neuropathic rats (rats with CCI) according to the protocol described in Example 2 above after anesthesia with pentobarbital sodium (100 mg / kg body weight ip). In both cases, the rats were adult Charles River CrI: CD BR rats weighing 400-500 g.
Les ganglions prélevés ont été incubés dans du milieu de EEAGLE modifié par le milieu DULBECCO (milieu DMEM) contenant 0,5 mg/ml de trypsine, 1 mg/ml de collagénase et 0,1 mg/ml d'ADNase pendant 90 minutes à 37°C. Un inhibiteur de trypsine de soja a été ensuite ajouté pour neutraliser la trypsine et la suspension résultante a été centrifugée à 1 000 g pendant une minute. Les neurones contenus dans le gâteau de centrifugation ont été mis en suspension dans le milieu DMEM contenant 10 % de sérum de veau, 10 % de sérum de cheval, 2 mM de glutamine, 0,1 mg/ml de pénicilline/streptomycine et 2,5 mg/ml d'ADNase et répartis dans des boîtes de Pétri (diamètre 35 mm) revêtues de Matrigel. Les neurones ont été ensuite incubés à 37°C en présence de 95 % de CO2 et 5 % d'O2 dans un milieu de base sous différentes conditions expérimentales :The lymph nodes removed were incubated in medium of EEAGLE modified with DULBECCO medium (DMEM medium) containing 0.5 mg / ml of trypsin, 1 mg / ml of collagenase and 0.1 mg / ml of DNAase for 90 minutes. 37 ° C. A soy trypsin inhibitor was then added to neutralize the trypsin and the resulting suspension was centrifuged at 1000 g for one minute. The neurons contained in the centrifuge cake were suspended in DMEM medium containing 10% calf serum, 10% horse serum, 2 mM glutamine, 0.1 mg / ml penicillin / streptomycin and 2.5 mg / ml DNase and distributed in Petri dishes (diameter 35 mm) coated with Matrigel. The neurons were then incubated at 37 ° C in the presence of 95% CO 2 and 5% O 2 in a basal medium under different experimental conditions:
- en présence du composé A (0,1 μM et 1 μM) ;in the presence of compound A (0.1 μM and 1 μM);
- en présence du facteur de croissance du tissu nerveux (NGF) (1 ng/ml et 50 ng/ml) ;in the presence of nerve growth factor (NGF) (1 ng / ml and 50 ng / ml);
- en l'absence de tout agent (rats témoins). Après 48 h d'incubation dans les conditions ci-dessus, les neurones cultivés ont été fixés dans du paraformaldéhyde 4 % pendant 5 à 10 minutes, lavés avec du tampon phosphate (PBS) et incubés immédiatement après avec un tampon bloquant (0,1 % de Tween 20 + 1,5 % de lait en poudre maigre dans du PBS à pH 4) pendant 30 minutes. Les cultures ont été ensuite incubées pendant une nuit à 40C avec une première solution d'anticorps (anticorps monodonaux de souris anti-neurofilament H et M commercialisés sous la dénomination Chemicon ® par la Société CHEMICON International Inc., Californie, USA) dans le tampon bloquant. Les cultures du jour suivant ont été lavées et incubées avec un second anticorps universel conjugué avec de la peroxydase (selon la technique ABC utilisant un complexe enzymatique avidine-biotine avec le kit commercialisé sous la dénomination Vectastain ® par la société VECTOR Laboratories à Burlingame, Californie, USA) et colorées avec une solution de 3,3'-diaminobenzidine (DAB). Les neurones ont été observés au microscope ZEISS et des photos ont été prises avec une caméra numérique Canon Power Shot G (4.0 mégapixel).- in the absence of any agent (control rats). After 48 h of incubation under the above conditions, the cultured neurons were fixed in 4% paraformaldehyde for 5 to 10 minutes, washed with phosphate buffer (PBS) and incubated immediately afterwards with a blocking buffer (0.1 % Tween 20 + 1.5% lean milk powder in PBS at pH 4) for 30 minutes. The cultures were then incubated overnight at 40 ° C. with a first antibody solution (anti-neurofilament mouse monodonal antibodies H and M marketed under the name Chemicon® by CHEMICON International Inc., California, USA) in the blocking buffer. The next day cultures were washed and incubated with a second universal peroxidase-conjugated antibody (according to the ABC technique using an avidin-biotin enzyme complex with the kit sold under the name Vectastain® by VECTOR Laboratories in Burlingame, California. , USA) and stained with a solution of 3,3'-diaminobenzidine (DAB). The neurons were observed under a ZEISS microscope and pictures were taken with a Canon Power Shot G (4.0 megapixel) digital camera.
Les explants DRG ont été mesurés sur chaque photographie d'une manière aléatoire en utilisant le logiciel SCION Image NIH (National Institute of Health Bethesda, Maryland USA).DRG explants were measured on each photograph in a random manner using SCION Image NIH software (National Institute of Health Bethesda, Maryland USA).
Un minimum de 15 explants DRG séparés ont été mesurés pour chaque boîte.A minimum of 15 separate DRG explants were measured for each box.
Dans chaque cas, la croissance maximale de neurites radiaux a été mesurée et convertie en unités métriques (élongation) par comparaison à une grille de calibration établie dans les mêmes conditions expérimentales. L'élongation des neurites DRF induite par NGF (exprimée en μm) dans les différentes conditions expérimentales est représentée sur la figure 1 :In each case, the maximum growth of radial neurites was measured and converted to metric units (elongation) compared to a calibration grid established under the same experimental conditions. The NGF-induced DRF elongation of neurites (expressed in μm) under the various experimental conditions is shown in FIG.
- rats témoins (sans CCI) ;- control rats (without CCI);
- rats témoins (sans CCI)+ NGF à 20 ng/ml ;control rats (without CCI) + NGF at 20 ng / ml;
- rats avec CCI ;- rats with CCI;
- rats avec CCI + NGF à 20 ng/ml ;rats with CCI + NGF at 20 ng / ml;
- au jour 0 (t = 0 jour de la création de la lésion chronique par constriction) ;- at day 0 (t = 0 days of the creation of the chronic lesion by constriction);
- au jour 3 et au jour 7 (t = 3 et t = 7, respectivement le 3eme et le 7eme jour après la création de la lésion chronique par constriction).- at day 3 and day 7 (t = 3 and t = 7, respectively the 3rd and 7th day after the establishment of the chronic constriction injury).
En condition nuisible, le NGF exogène provoque une augmentation de l'élongation des neurites DRG qui est maximale le 3ème jour après la lésion. Le 7eme jour, le NGF exogène est incapable de provoquer une augmentation supplémentaire de l'élongation des neurites DRG, ceci étant probablement dû à l'activation des cellules Schwann (processus de régénération).In adverse conditions, the exogenous NGF leads to an increase in elongation DRG neurites which is the maximum day 3 after injury. The 7 th day, the exogenous NGF is unable to cause further increase in neurite elongation DRG, probably due to activation of Schwann cells (regeneration process).
On a ensuite mesuré le % d'élongation des neurites après administration du composé A à la concentration 10"6 M ou 10"7 M. Les pourcentages d'effet sont calculés par rapport à l'effet dû au NGF. Les résultats obtenus sont rassemblés dans le tableau 4 ci-après.The percent neurite elongation was then measured after administration of compound A at the concentration of either 10 -6 M or 10 -7 M. The effect percentages are calculated relative to the effect due to NGF. The results obtained are summarized in Table 4 below.
Tableau 4 : Elongation des neuritesTable 4: Neurite elongation
Figure imgf000011_0001
Figure imgf000011_0001
* p < 0,05,* p <0.05,
** p < 0,01 ANOVA test Duncan / témoins n = 3-5 TEST 4 : Effet du composé A sur l'hypertrophie bénigne de la prostate** p <0.01 ANOVA Duncan test / controls n = 3-5 TEST 4: Effect of compound A on benign prostatic hypertrophy
Une hypertrophie bénigne de la prostate (HBP) a été provoquée chez 10 rats mâles CD par administration de testostérone à raison de 18,75 mg par jour par voie sous-cutanée pendant 24 jours. Le composé A a été ensuite administré par voie orale en combinaison avecBenign prostatic hypertrophy (BPH) was induced in 10 male CD rats by administration of 18.75 mg testosterone per day subcutaneously for 24 days. Compound A was then administered orally in combination with
NaOH 10% +Tween 80 5% +HCI IN 9,8%+H2O comme véhicule (5 ou 10 mg de composé A par ml de véhicule) à raison de 10 ou 30 mg/kg de poids corporel et par jour pendant 24 jours. On a ensuite mesuré le volume et le poids de la prostate totale et de la prostate ventrale. Les résultats obtenus sont reportés sur les figures 2a et 2b (poids) et les figures 3a et 3b (volume) ainsi que ceux obtenus avec des rats mâles témoins chez lesquels une hypertrophie bénigne de la prostate a été provoquée selon le mode opératoire ci-dessus mais auxquels on a administré uniquement le véhicule.10% NaOH + Tween 80 5% + HCl IN 9.8% + H 2 O as vehicle (5 or 10 mg of compound A per ml vehicle) at a rate of 10 or 30 mg / kg of body weight and per day during 24 days. The volume and weight of the total prostate and ventral prostate were then measured. The results obtained are shown in FIGS. 2a and 2b (weight) and FIGS. 3a and 3b (volume) as well as those obtained with male control rats in which a benign prostatic hypertrophy was provoked according to the above procedure. but to which only the vehicle has been administered.
TEST 5 : mesure isométrique On a réalisé le test ex vivo ci-après sur le lobe ventral de la prostate de rats chez lesquels une hypertrophie bénigne a été induite selon le mode opératoire décrit au test 4.TEST 5: Isometric measurement The following ex vivo test was performed on the ventral lobe of the prostate of rats in which benign hypertrophy was induced according to the procedure described in test 4.
La prostate a été rapidement excisée chez des rats mâles Crl:CD® Charles River) d'un poids de 300-35Og et mise dans le tampon bicarbonate de Krebs (NaCI 118,4 mM; KCI 4,7 mM, CaCI2 2,5 mM, KH2PO4 1,2 mM, NaHCO3 25 mM, glucose 11,7 mM).The prostate was rapidly excised in male rats Crl: CD ® Charles River) weighing 300-35Og and placed in Krebs bicarbonate buffer (118.4 mM NaCl, 4.7 mM KCl, CaCl 2 2, 5mM, 1.2mM KH 2 PO 4 , 25mM NaHCO 3 , 11.7mM glucose).
La prostate a été débarrassée des tissus environnants et la partie ventrale a été isolée et coupée en deux lobes. Chaque lobe a été monté longitudinalement dans une chambre d'organe isolé de 20 ml pour enregistrement de la tension isométrique, mis dans le tampon bicarbonate de Krebs à 37°C, oxygéné avec 95% d'O2 - 5% de CO2 et étiré jusqu'à une tension passive de 250 mg. Après une période d'équilibration durant laquelle le lobe a été lavé avec le tampon Krebs tel que défini ci-dessus 4 fois pendant 10 minutes, la courbe de réponse à la noradrénaline (1 nM - 0,1 mM) a été obtenue en présence de 0,5 μM de DMI (desméthylimipramine, Ciba) et 30 μM d'hémisuccinate dtiydrocortisone (Flebocortid ®, Lepetit). Les changements de forces isométriques ont été enregistrés en utilisant le système d'acquisition des données PowerLab (AD Instruments Pty Ltd, Castle HiII, Australie). La génération de la force isométrique a été mesurée comme étant le pourcentage de contraction induite par KCI 40 mM (100% de contraction maximale tissulaire) ajouté à la fin de l'expérimentation après la concentration la plus élevée de noradrénaline.The prostate was cleared of surrounding tissue and the ventral portion was isolated and cut into two lobes. Each lobe was mounted longitudinally in a 20 ml isolated organ chamber for recording the isometric tension, put in Krebs bicarbonate buffer at 37 ° C, oxygenated with 95% O 2 - 5% CO 2 and stretched to a passive voltage of 250 mg. After an equilibration period during which the lobe was washed with Krebs buffer as defined above 4 times for 10 minutes, the norepinephrine response curve (1 nM - 0.1 mM) was obtained in the presence 0.5 μM of DMI (desmethylimipramine, Ciba) and 30 μM of hydrocortisone hemisuccinate (Flebocortid®, Lepetit). The isometric force changes were recorded using the PowerLab data acquisition system (AD Instruments Pty Ltd, Castle HiII, Australia). The generation of isometric force was measured as the percentage of contraction induced by 40 mM KCI (100% maximal tissue contraction) added at the end of the experiment after the highest concentration of norepinephrine.
Les résultats sont exprimés en EC50 (concentration produisant 50% de l'effet maximal).The results are expressed in EC50 (concentration producing 50% of the maximum effect).
Les courbes de réponse à la noradrénaline sont représentées sur la figure 4.The norepinephrine response curves are shown in Figure 4.
Chez les rats ayant une hypertension bénigne provoquée par la testostérone (rats témoins + HBP), la courbe de réponse à la noradrénaline montre un décalage vers la droite par rapport aux rats témoins. Ce décalage est significatif d'une diminution de la capacité de contraction tissulaire.In rats with benign testosterone-induced hypertension (control + BPH rats), the norepinephrine response curve showed a shift to the right compared with the control rats. This shift is indicative of a decrease in tissue contraction capacity.
Le composé A administré à raison de 10 ou 30 mg/kg de poids corporel a montré une capacité de contraction normale à la noradrénaline. Le composé A restaure donc la contractibilité de l'échantillon de prostate testé.Compound A administered at 10 or 30 mg / kg body weight showed a normal contraction capacity to norepinephrine. Compound A thus restores the contractibility of the tested prostate sample.
Ainsi, les exemples montre que le composé A, ses esters, ses sels pharmaceutiquement acceptables et/ou leurs solvates sont utiles pour le traitement ou pour la préparation d'un médicament destiné au traitement des douleurs chroniques, des douleurs neuropathiques, des neuropathies notamment de la neuropathie diabétique, de l'inflammation, notamment de l'inflammation asthmatique bronchopulmonaire ou inflammation allergique bronchopulmonaire, du syndrome du colon irritable, de la maladie de Crohn, de l'inflammation des intestins et peuvent être utilisés pour la préparation d'un médicament destiné à un tel traitement.Thus, the examples show that the compound A, its esters, its pharmaceutically acceptable salts and / or their solvates are useful for the treatment or for the preparation of a medicament intended for the treatment of chronic pain, neuropathic pain, neuropathies, especially neuropathies. Diabetic neuropathy, inflammation, including bronchopulmonary asthmatic inflammation or bronchopulmonary allergic inflammation, irritable bowel syndrome, Crohn's disease, inflammation of the intestines and may be used for the preparation of a drug intended for such treatment.
La présente invention, selon un autre de ses aspects, concerne également une méthode de traitement des pathologies ci-dessus indiquées qui comprend l'administration, à un patient, d'une dose efficace du composé A ses esters, ses sels pharmaceutiquement acceptables et/ou leurs solvates. The present invention, according to another of its aspects, also relates to a method of treatment of the pathologies indicated above which comprises the administration to a patient of an effective dose of the compound A its esters, its pharmaceutically acceptable salts and / or or their solvates.

Claims

REVENDICATIONS
1. Utilisation de l'acide 5-(4-chlorophényl)-l-(2,4-dichlorophényl)-4-méthylpyrazole- 3-carboxylique, ses esters, ses sels pharmaceutiquement acceptables et leurs solvates pour la préparation d'un médicament destiné au traitement de la douleur, des neuropathies, de l'inflammation ou de l'hypertrophie bénigne de la prostate.1. Use of 5- (4-chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its esters, pharmaceutically acceptable salts and solvates for the preparation of a medicament for the treatment of pain, neuropathy, inflammation or benign prostatic hypertrophy.
2. Utilisation selon la revendication 1 caractérisée en ce que la douleur est la douleur chronique.2. Use according to claim 1 characterized in that the pain is chronic pain.
3. Utilisation selon la revendication 1 caractérisée en ce que la douleur est la douleur neuropathique.3. Use according to claim 1 characterized in that the pain is neuropathic pain.
4. Utilisation selon la revendication 1 caractérisée en ce que la neuropathie est la neuropathie du diabétique.4. Use according to claim 1 characterized in that the neuropathy is the neuropathy of the diabetic.
5. Utilisation selon la revendication 1 caractérisée en ce que l'inflammation est l'inflammation asthmatique bronchopulmonaire ou inflammation allergique bronchopulmonaire, le syndrome du colon irritable, la maladie de Crohn ou l'inflammation des intestins.5. Use according to claim 1 characterized in that the inflammation is bronchopulmonary asthmatic inflammation or bronchopulmonary allergic inflammation, irritable bowel syndrome, Crohn's disease or inflammation of the intestines.
6. Utilisation de l'acide 5-(4-chlorophényl)-l-(2,4-dichlorophényl)-4-méthylpyrazole- 3-carboxylique, son ester, son sel pharmaceutiquement acceptable et son solvate pour la préparation de médicaments utile pour le traitement de l'hypertrophie bénigne de la prostate.6. Use of 5- (4-chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its ester, pharmaceutically acceptable salt and solvate for the preparation of medicaments useful for treatment of benign prostatic hypertrophy.
7. Acide 5-(4-chlorophényl)-l-(2,4-dichlorophényl)-4-méthylpyrazole-3-carboxylique, son ester, son sel pharmaceutiquement acceptable et son solvate pour le traitement de la douleur, des neuropathies, de l'inflammation ou de l'hypertrophie bénigne de la prostate. 7. 5- (4-Chlorophenyl) -1- (2,4-dichlorophenyl) -4-methylpyrazole-3-carboxylic acid, its ester, its pharmaceutically acceptable salt and its solvate for the treatment of pain, neuropathies, inflammation or benign prostatic hypertrophy.
PCT/FR2008/001095 2007-07-26 2008-07-24 5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxylic acid the pharmaceutically-acceptable esters and salts and solvates thereof for use as medicament WO2009044006A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR0705483A FR2919184A1 (en) 2007-07-26 2007-07-26 Use of 5-(4-chlorophennyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxylic acid as a medicament for treating e.g. chronic pain, neuropathic pain, inflammation, benign hypertrophy of the prostate and diabetic neuropathy
FR0705483 2007-07-26

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WO2009044006A2 true WO2009044006A2 (en) 2009-04-09
WO2009044006A3 WO2009044006A3 (en) 2009-06-11

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0656354A1 (en) * 1993-12-02 1995-06-07 Sanofi Substituted N-piperidino 3-pyrazolecarboxamide
US5624941A (en) * 1992-06-23 1997-04-29 Sanofi Pyrazole derivatives, method of preparing them and pharmaceutical compositions in which they are present
WO2003088968A1 (en) * 2002-04-15 2003-10-30 Research Triangle Institute Compounds having unique cb1 receptor binding selectivity and methods for their production and use
FR2864958A1 (en) * 2004-01-12 2005-07-15 Sanofi Synthelabo New 1,5-diphenyl-3-sulfonamidomethyl-1H-pyrazole derivatives are cannabinoid receptor antagonists used for treating e.g. eating disorders, gastrointestinal disorders, inflammation and immunological disorders
WO2007009701A2 (en) * 2005-07-15 2007-01-25 Laboratorios Del Dr. Esteve, S.A. Use of substituted pyrazole compounds and combinations thereof for the treatment of the metabolic syndrome

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5624941A (en) * 1992-06-23 1997-04-29 Sanofi Pyrazole derivatives, method of preparing them and pharmaceutical compositions in which they are present
EP0656354A1 (en) * 1993-12-02 1995-06-07 Sanofi Substituted N-piperidino 3-pyrazolecarboxamide
WO2003088968A1 (en) * 2002-04-15 2003-10-30 Research Triangle Institute Compounds having unique cb1 receptor binding selectivity and methods for their production and use
FR2864958A1 (en) * 2004-01-12 2005-07-15 Sanofi Synthelabo New 1,5-diphenyl-3-sulfonamidomethyl-1H-pyrazole derivatives are cannabinoid receptor antagonists used for treating e.g. eating disorders, gastrointestinal disorders, inflammation and immunological disorders
WO2007009701A2 (en) * 2005-07-15 2007-01-25 Laboratorios Del Dr. Esteve, S.A. Use of substituted pyrazole compounds and combinations thereof for the treatment of the metabolic syndrome

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FR2919184A1 (en) 2009-01-30
WO2009044006A3 (en) 2009-06-11

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