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  • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
  • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
  • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D215/20—Oxygen atoms
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  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/47—Quinolines; Isoquinolines
  • A61K31/4704—2-Quinolinones, e.g. carbostyril
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• • A—HUMAN NECESSITIES
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  • A61P19/00—Drugs for skeletal disorders
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• • A—HUMAN NECESSITIES
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  • A61P19/00—Drugs for skeletal disorders
  • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
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• • A—HUMAN NECESSITIES
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  • A61P25/00—Drugs for disorders of the nervous system
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• • A—HUMAN NECESSITIES
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  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
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• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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• • A—HUMAN NECESSITIES
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  • A61P5/00—Drugs for disorders of the endocrine system
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• • A—HUMAN NECESSITIES
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  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D215/00—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems
  • C07D215/02—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom
  • C07D215/16—Heterocyclic compounds containing quinoline or hydrogenated quinoline ring systems having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen atoms or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
  • C07D215/48—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
  • C07D215/54—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen attached in position 3
  • C07D215/56—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen attached in position 3 with oxygen atoms in position 4

Definitions

• the present invention relates to compounds derived from quinolinone capable of modulating the activity, in particular of inducing differentiation, of stem and progenitor cells; these compounds are useful for the treatment of disorders related to a lack of cell differentiation; the invention also relates to novel compounds among these quinolinone derivatives and to pharmaceutical compositions containing them.
• the repair of damaged tissue following disease, trauma or age increasingly uses stem cells or progenitors that retain the ability to differentiate to different cell types. These cells constitute a reservoir capable of renewing tissues in order to restore biological functions.
• Mesenchymal stem cells for example, can yield osteoblasts, chondrocytes, adipocytes, or stromal cells that support hematopoiesis.
• disorders resulting from a defect in cell differentiation are those related to dysfunction of osteoblast differentiation.
• Bone is continually renewed during life by a complex process involving resorption by osteoclasts and osteoblast formation.
• the precursors of osteoblasts are pluripotent cells also called mesenchymal stem cells.
• mesenchymal stem cells the precursors of osteoblasts are pluripotent cells also called mesenchymal stem cells.
• the mechanisms that allow the differentiation of these cells to the osteoblastic lineage are complex and are of great importance in understanding the development of bone.
• the identification of molecules that induce osteoblast differentiation and stimulate their osteogenic activity would represent a therapeutic avenue in the treatment of bone diseases.
• osteoporosis is characterized by excessive fragility of the skeleton due to a decrease in bone mass and alteration of bone microarchitecture.
• the strength of the bone results from a balance between the action of two types of bone cells: osteoblasts that solidify the bone and osteoclasts (responsible for bone resorption) which weaken them.
• Dominant activity of osteoclasts leads to osteoporosis, which can result either in bone deficiency at the end of growth or excessive bone loss in old age.
• the prevention of osteoporosis can be done by reducing a precursory physiological phenomenon, osteopenia (a decrease in bone density) which, before osteoporosis, can lead to bone rarefaction disorders and the weakening of the tissue. bony.
• this disease is also called "glass bone disease” includes diseases characterized by excessive bone fragility, due to a congenital defect in the development of coUagene fibers connective tissue that forms the frame of the bone. All types are characterized by extremely fragile bones, the most typical sign of the disease;
• osteomalacia which corresponds to a bone decalcification induced by a lack of mineralization (lack of calcium and phosphate ions) of the skeletal protein matrix;
• osteonecrosis which covers conditions defined by the death of bone cells
• Renewal of bone tissue may also be necessary in situations where bone repair is attempted such as fractures, plastic surgery or placement of implants, including dental implants.
• Osteoblastic differentiation is influenced by multiple signaling pathways including, for example, TGF- ⁇ (transforming growth factor B1), Hedgehog (Hh), Wnt, FGF (fibroblast growth factors), IGF1 ( insulin-like growth factor 1) or bone morphogenetic proteins (BMPs) (Centrella et al., 1994, Yamaguchi et al., 2000, van der Horst et al., 2003, Fromigue et al., 2004 and Hu et al., 2005).
• TGF- ⁇ transforming growth factor B1
• Hh Hedgehog
• Wnt fibroblast growth factors
• FGF fibroblast growth factors
• IGF1 insulin-like growth factor 1
• BMPs bone morphogenetic proteins
• TGF- ⁇ has also been described as a major regulator of the balance of activity between osteoclasts and osteoblasts. Recently, pharmacological inhibitors of its receptor have shown a stimulating activity on osteoblasts and an inhibitory effect on osteoclasts (Mohammad et al., 2009).
• IGF1 The role of IGF1 has been well studied, but the use of recombinant human IGF1, despite its influence on bone metabolism, has certain disadvantages. It does not specifically target the skeleton and causes side effects that limit its use for bone diseases.
• the Hedgehog signaling molecule plays a fundamental role in the morphogenesis of many tissues, including bone, as well as in cell proliferation, and is thought to be involved in tissue maintenance and repair in adults (see Ingham's reviews). McMahon 2001, Wechsler-Reya and Scott 2001, Marti and Bovolenta 2002, Lum and Beachy 2004, Varjosalo and Taipale 2008).
• Hedgehog pathway allows the induction of osteogenesis in different models.
• Several agonist molecules have been studied: - Hedgehog proteins and derived polypeptides that stimulate osteoblast differentiation by acting on the Patched protein (Spinella-Jaegle et al., 2001, Guan et al., 2009);
• Oxysterol derivatives (Corcoran and Scott 2006, Amantea et al., 2008) that induce osteoblast differentiation and bone formation (Aghaloo et al., 2007, Dwyer et al., 2007, Yu et al., 2008).
• the inventors have identified compounds of general formula (I) capable of inducing cell differentiation, in particular osteoblastic differentiation; these compounds therefore represent novel agents stimulating the differentiation of stem cells or progenitor cells towards osteoblasts or other cell types.
• stem cell an undifferentiated, embryonic or adult cell capable of giving specialized cells by cell differentiation and which can be renewed practically indefinitely.
• Adult stem cells will preferably be used.
• the progenitor cells are pluripotent adult cells, that is to say, whose differentiation can lead to several cell types.
• cells are involved in osteoblastic differentiation - they are then referred to as "cells with osteoblastic phenotype" - when they produce alkaline phosphatase.
• Such cells are sufficiently engaged in osteoblast differentiation for the continuation of their incubation in the nutrient medium and / or their implantation to allow at least a portion of said cells to advance in differentiation, to terminal differentiation (with production a mineralized extracellular matrix).
• Highlighting properties characteristics of these cells ie production of alkaline phosphatase
• X positioned in ortho, meta or para, represents -H, -OH, -NH 2 , a halogen atom, preferably chlorine or bromine, an alkyl radical consisting of a carbon chain of 1 to 10 atoms of carbon, linear or branched, an alkoxy radical (of formula -O-alkyl where the alkyl group is as defined above), a cycloalkyl of 3 to 8 carbon atoms or an aryl group
• - W represents -H, -OH, -NH 2 or a halogen atom, preferably chlorine or bromine;
• R 1, R 2 and R 3 identical or different and independently of each other, represent:
• alkyl group consisting of a linear or branched, optionally unsaturated, carbon chain of 1 to 10 carbon atoms with one or more double or triple bonds, optionally substituted with one or more heteroatoms such as O and S, with one or more atoms; halogens or with one or more aryl or heteroaryl groups, preferably a pyridine group; or
• a cycloalkyl of 3 to 8 carbon atoms optionally substituted by an alkyl radical consisting of a carbon chain of 1 to 10 atoms of carbon, linear or branched or an alkoxy radical (of formula -O-alkyl where the alkyl group is as defined above);
• R 4 , R 5 , R 6 and R 7, which are identical or different and independently of each other, are chosen from -H, -Cl, -Br, -I, -CN, -NO 2 , an alkyl radical consisting of carbon chain of 1 to 10 carbon atoms, linear or branched, an alkoxy radical (of formula -O-alkyl where the alkyl group is as defined above) or a cycloalkyl of 3 to 8 carbon atoms, for example a cyclopropyl, cyclopentyl, cyclohexyl;
• R 3 and R 4 may be fused to form, with the carbon and adjacent nitrogen atoms of the quinoline ring which carries them, a 5- or 6-membered ring;
• alkyl is understood to mean a linear or branched, saturated hydrocarbon aliphatic group having from 1 to 10 carbon atoms, preferably from 3 to 8 carbon atoms.
• branched means that at least one lower alkyl group of 1 to 6 carbon atoms, such as methyl or ethyl, is carried by a linear alkyl chain.
• Halogen atom means a bromine, chlorine, iodine or fluorine atom; the bromine and chlorine designations being preferred.
• aryl group any functional group or substituent derived from at least one aromatic ring; mention may be made of phenyl, benzylcyclobutene, pentalene, naphthalene, benzylphenyl and anthracene groups.
• heteroaryl group any functional group or substituent derived from at least one aromatic ring as defined above and containing at least one heteroatom selected from P, S, O and N; among the heteroaryl groups, mention may be made of furan, pyridine, pyrrole, thiophene, imidazole, pyrazole, oxazole, isoxazole, thiazole, pyridine, pyrazine, pyrimidine, pyridazine, benzofuran, isobenzofuran, indole, isoindole, benzothiophene, benzo [c] groups; thiophene, benzimidazole, indazole, benzoxazole, benzisoxazole, benzothiazole, quinoline, isoquinoline, quinoxaline, quinazoline, cinnoline, purine, and acridine.
• the compounds of general formula (I) are such that the radical R 3 is an alkyl radical of 3 to 8 carbon atoms such as the hexyl radical; the radical R 2 is a hydrogen atom or an alkyl radical of 1 to 3 carbon atoms; that the radicals R 4 , R 5 , R 6 and R 7 represent a hydrogen atom or an alkyl radical of 1 to 3 carbon atoms.
• the compounds of general formula (I) according to the invention may be prepared according to the synthesis scheme shown in FIG.
• aniline is condensed with triethyl methane tricarboxylate under microwave irradiation to obtain quinolinones carboxylates; then the quinolones are condensed with 4-tert-butylaniline under microwave irradiation to give carboxamidoquinolinone esters; the next step is to saponify the ester function of these compounds to give an acid starting point for various modifications as illustrated in the experimental examples.
• the compounds of general formula (I) according to the invention possess the property of inducing the differentiation of mesenchymal cells into osteoblasts.
• these compounds are of interest for the preparation of implants obtained from autologous cells which will have been differentiated into osteoblates; these implants are useful for correcting bone loss resulting from injuries and / or surgical operations.
• the compounds of general formula (I) are therefore of particular interest for the treatment of diseases caused by the dysregulation of the function or differentiation of osteoblasts and / or by the functional imbalance between osteoblasts and osteoclasts.
• these compounds are useful for the treatment of osteoporosis; and disorders such as skeletal fragility and / or bone scarcity and / or bone tissue fragility resulting from osteopenia; osteogenesis imperfecta; hypercalcemia; hyperparathyroidism; osteomalacia; osteonecrosis; Paget's disease of bone (deforming osteitis); rheumatoid arthritis; inflammatory arthritis; osteomyelitis; periodontitis; bone metastases.
• disorders such as skeletal fragility and / or bone scarcity and / or bone tissue fragility resulting from osteopenia; osteogenesis imperfecta; hypercalcemia; hyperparathyroidism; osteomalacia; osteonecrosis; Paget's disease of bone (deforming osteitis); rheumatoid arthritis; inflammatory arthritis; osteomyelitis; periodontitis; bone metastases.
• tissue dysfunction to induce the formation, regeneration, repair and / or increase of tissue activity
• tissue activity such as but not limited to: nerve tissue [central (brain) and peripheral nervous system (neurons sensory, motor, sympathetic], bone, cartilage, testes, liver, spleen, intestine, pancreas, kidneys, smooth and skeletal muscles, heart, lungs, skin and system hair, mucous membranes, blood cells and cells of the immune system, by way of non-limiting example of these pathologies, mention may in particular be made of neuropathies and associated neuromuscular diseases, diabetes, alopecia, burns, ulcerations (skin and mucous membrane) and spermatogenic disorders.
• the present invention also relates to a process for preparing osteoblast cells from stem cells or progenitor cells, such as mesenchymal cells, comprising the step of incubating said cells for a sufficient time in a liquid nutrient medium allowing the development of said cells, said nutrient medium containing in solution at least one compound of general formula (I).
• said cells are incubated in the culture medium, under standard conditions allowing their development, that is to say not only their survival but also their proliferation and / or their differentiation.
• Standard conditions human cell culture are known: for example temperature of about 37 ° C; atmosphere air-COss 95: 5; pH close to neutrality.
• the culture medium used is a conventional liquid nutrient medium containing the ingredients necessary for the development of mammalian cells. These ingredients are known. They are mainly mineral salts (in particular Na, K, Mg, Ca and possibly Cu, Fe, Zn), amino acids, vitamins and sources of carbon (for example glucose).
• a nutrient medium such as the minimum essential MEM medium of EAGLE supplemented with fetal calf serum or, preferably, with autologous human serum may be used.
• These culture media may be supplemented with osteoinductive factors, such as BMP factors, but, as indicated above, these factors are not necessary to induce osteoblastic differentiation of human cells used in the method of the invention. It is therefore possible to use media free of osteoinductive factors.
• osteoinductive factors such as BMP factors
• the culture media used are supplemented with osteopromotor factors such as ascorbic acid and beta-glycerophosphates (for example sodium or calcium).
• osteopromotor factors such as ascorbic acid and beta-glycerophosphates (for example sodium or calcium).
• the present invention also relates to the novel compounds of general formula (Ia):
• X positioned in ortho, meta or para, represents -H, -OH, -NH 2 , a halogen atom, preferably chlorine or bromine, an alkyl radical consisting of a carbon chain of 1 to 10 carbon atoms; , linear or branched, an alkoxy radical (from formula -O-alkyl where the alkyl group is as defined above), a cycloalkyl of 3 to 8 carbon atoms or an aryl group;
• - W represents -H, -OH, -NH 2 or a halogen atom, preferably chlorine or bromine;
• R 1 and R 3 which are identical or different, and independently of each other, represent an alkyl group consisting of a linear or branched carbon chain of 3 to 10 carbon atoms, optionally unsaturated by one or more double or triple bonds, optionally substituted with one or more heteroatoms such as O and S, with one or more halogen atoms or with one or more aryl or heteroaryl groups, preferably a pyridine group;
• R 4 , R 5 , R 6 and R 7, which are identical or different and independently of each other, are chosen from -H, -Cl, -Br, -I, -CN, -NO 2 , an alkyl radical consisting of carbon chain of 1 to 10 carbon atoms, linear or branched, an alkoxy radical (of formula -O-alkyl where the alkyl group is as defined above) or a cycloalkyl of 3 to 8 carbon atoms, for example a cyclopropyl, cyclopentyl, cyclohexyl;
• R 3 and R 4 may be fused to form, with the carbon and adjacent nitrogen atoms of the quinoline ring which carries them, a 6-membered ring;
• the preferred radicals R 5 and R 7 are chosen, independently of one another, from -H, -Cl, -Br, -CN, an alkyl radical or an alkoxy radical.
• the dosage will vary depending on the condition to be treated, the route and timing of administration, and the nature and weight of the subject to be treated (human or animal).
• the present invention also relates to pharmaceutical compositions characterized in that they comprise, as active principle, at least one compound of general formula (Ia) according to the invention, and at least one pharmaceutically acceptable excipient.
• the compound or compounds of general formula (Ia) are preferably used in an amount for administering unit doses of between 1 mg and 2 g approximately.
• the form of the drug or pharmaceutical composition e.g., solution, suspension, emulsion, tablets, capsules, suppositories, etc.
• the form of the drug or pharmaceutical composition e.g., solution, suspension, emulsion, tablets, capsules, suppositories, etc.
• the form of the drug or pharmaceutical composition will depend on the chosen route of administration.
• the drug or the pharmaceutical composition can be administered by any suitable route, for example by oral, anal, local, systemic, intravenous, intramuscular or mucosal route, or by using a patch, or in encapsulated form, or immobilized on liposomes, microparticles, microcapsules, and the like.
• excipients suitable for oral administration talc, lactose, starch and its derivatives, cellulose and its derivatives, polyethylene glycols, acid polymers acrylic, gelatin, magnesium stearate, animal, vegetable or synthetic fats, paraffin derivatives, glycols, stabilizers, preservatives, anti-oxidants, wetting agents, anti-caking agents, dispersants , emulsifiers, taste modifying agents, penetration agents, solubilization agents, etc.
• FIG. 1 shows the synthesis scheme of the compounds of general formula (I).
• Figure 2 illustrates the histochemical detection of alkaline phosphatase induced following the differentiation of mesenchymal stem cells under the action of the compounds of general formula (I).
• the cells were cultured in the presence of 10 ⁇ l of the indicated compounds 1, 4, 6 or 8 or the solvent (control) for 6 days in the usual culture medium.
• the cells were then stained by histochemistry to reveal the presence of alkaline phosphatase (red marking appearing as darker spots) marker of osteogenesis and then photographed.
• Figure 3 illustrates the effect of compound 1 on the enzymatic activity of alkaline phosphatase indicating the effect of this compound on the differentiation of C3H10T1 / 2 mesenchymal stem cells.
• the differentiation induced by compound 1 is measured by the enzymatic activity of alkaline phosphatase (OD at 415 nm) (round).
• OD at 415 nm the enzymatic activity of alkaline phosphatase
• FIG. 4 Differentiation of C3H10T1 / 2 Cells by Compound 2.
• the response to Compound 2 was evaluated under the same experimental conditions as those used for compound 1.
• the differentiation of the cells obtained in the same Experiment with compound 1 (10 ⁇ ) is indicated by a diamond.
• Figure 5 Differentiation of C3H10T1 / 2 cells induced by compound 3
• Figure 6 Differentiation of C3H10T1 / 2 cells induced by compound 4.
• Figure 7 Differentiation of C3H10T1 / 2 cells induced by compound 5.
• Figure 8 Differentiation of C3H10T1 / 2 cells induced by compound 6.
• N-hexylaniline 355 mg, 2 mmol
• a solution of triethyl ester methanetricarboxylic acid (1.35 ml, 6.4 mmol).
• the effect of the compounds of general formula (I) according to the invention on the stimulation of osteogenesis was determined in vitro by analysis of the differentiation of the pluripotent mesenchymal cell line C3H10T1 / 2.
• the compounds of formula (I) to be tested were dissolved in dimethylsulfoxide (DMSO) to a concentration of 2.5 mM, and then stored at a temperature of -20 ° C until use.
• DMSO dimethylsulfoxide
• the pluripotent fibroblast cell line C3H10T1 / 2 was cultured under the conditions recommended by ATCC in DMEM culture supplemented with 10% fetal calf serum at a temperature of 37 ° C under a 5% CO 2 atmosphere. 24 hours after seeding, stimulation of the cells was carried out for 6 days in the presence of the compounds of general formula (I) directly diluted in the culture medium. Activation by these compounds causes differentiation of the cell line to the osteoblast lineage and allows them to express alkaline phosphatase. This is then detected by histochemistry or enzymatic assay.
• cells are cultured in 6-well plates containing a 1H treated glass coverslip at 0.05 mg / ml polyD-Lysine.
• the C3H10T1 / 2 cells are seeded at a density of 150,000 cells per well.
• the compounds were applied at a concentration of 10 ⁇ .
• the SIGMA Staining Kit (85L-3R) was used following the protocol described. Briefly, after fixing the cells for 30 seconds with a solution of citrate / acetone (2-3) and then rinsing with double-distilled water, the coloration is carried out for 30 minutes in the presence of a solution of Fast-Violet / Naphtol shelter from the light. The slides are then rinsed thoroughly with distilled water and then mounted in an aqueous medium before photographing on a DMRXA2 microscope (Leica Microsystems). Cells expressing alkaline phosphatase are stained red.
• the cells were washed in cold phosphate buffer (PBS) and then lysed by sonication at 4 ° C. in 50 ⁇ l of a solution containing 0.9% NaCl and 0.2% Triton X-100.
• the measurement of the alkaline phosphatase activity in the lysates thus obtained was then carried out according to the method described by Pepinsky et al. (Pepinsky et al 1998).
• reaction buffer 200 mM Tris-HCl, pH 10.5, 0.4 M of 2-amino-2-methylpropanol and 8 mM of MgCl 2
• substrate 4 mM P - Nitrophenyl phosphate disodium
• the lysates were incubated at 37 ° C for 60 min, then the optical density (OD) was measured at a wavelength of 415 nm.
• OD optical density
• the activity of the compound SAG, activator of osteogenesis by action on the hedgehog pathway was tested under the same conditions.
• GraphPad Prism 4® software was used to plot the curves and determine the effective concentrations 50 (EC 50 ).
• Stimulation of osteogenesis in C3H10T1 / 2 pluripotent cells was assessed by observing the induction of alkaline phosphatase (AP). After 6 days of differentiation in the presence of 10 ⁇ l of compounds of formula (I), the expression of PA was detected by histochemical staining. By way of example, an increase in the number of labeled cells is observed in the presence of compounds 1, 4, 6 and 8 with greater or lesser intensities (compound 4> compound 1> compound 8> compound 6). No solvent-treated cells (DMSO) express AP at a level detectable by this method.
• DMSO solvent-treated cells
• FIG. 3 shows a representative curve of compound 1 produced in parallel with that of the smoothened SAG receptor agonist. The latter gives a maximum stimulation lower than the compound 1 while their affinities are close.
• Table I Affinity and maximum activation of the compounds of formula (I) on the differentiation of C3H10T1 / 2 multipotent cells.
• the effective concentration (EC 50 ) of the compounds on the differentiation is expressed in ⁇ .
• the maximum stimulation is expressed as a percentage of that obtained with compound 1 in the same experiment.
• the data correspond to the average of 2 to 5 independent experiments.
• Hedgehog Signaling Identification and Characterization of Smoothened Agonists and Antagonists. J Biol 1 (2): 10.

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• Medicines Containing Material From Animals Or Micro-Organisms (AREA)
• Micro-Organisms Or Cultivation Processes Thereof (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

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• 2010
  • 2010-11-16 FR FR1004445A patent/FR2967353B1/fr active Active
• 2011
  • 2011-11-15 JP JP2013538319A patent/JP5914509B2/ja not_active Expired - Fee Related
  • 2011-11-15 EP EP11794858.8A patent/EP2640388B1/fr not_active Not-in-force
  • 2011-11-15 WO PCT/IB2011/055096 patent/WO2012066478A1/fr not_active Ceased
  • 2011-11-15 US US13/885,089 patent/US9061997B2/en not_active Expired - Fee Related
  • 2011-11-15 ES ES11794858.8T patent/ES2651015T3/es active Active
  • 2011-11-15 CA CA2817388A patent/CA2817388C/fr not_active Expired - Fee Related

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