WO1997015308A1 - Compositions et procedes pour le traitement des deficits osseux - Google Patents

Compositions et procedes pour le traitement des deficits osseux Download PDF

Info

Publication number
WO1997015308A1
WO1997015308A1 PCT/US1996/017019 US9617019W WO9715308A1 WO 1997015308 A1 WO1997015308 A1 WO 1997015308A1 US 9617019 W US9617019 W US 9617019W WO 9715308 A1 WO9715308 A1 WO 9715308A1
Authority
WO
WIPO (PCT)
Prior art keywords
bone
substituted
compounds
composition
ofthe
Prior art date
Application number
PCT/US1996/017019
Other languages
English (en)
Other versions
WO1997015308A9 (fr
Inventor
Charles Petrie
Mark W. Orme
Nand Baindur
Kirk G. Robbins
Scott M. Harris
Maria Kontoyianni
Laurence H. Hurley
Sean M. Kerwin
Gregory R. Mundy
Original Assignee
Zymogenetics, Inc.
Osteoscreen, Inc.
University Of Texas At Austin
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Zymogenetics, Inc., Osteoscreen, Inc., University Of Texas At Austin filed Critical Zymogenetics, Inc.
Priority to BR9611210-7A priority Critical patent/BR9611210A/pt
Priority to JP09516761A priority patent/JP2000513324A/ja
Priority to KR1019980702947A priority patent/KR19990067010A/ko
Priority to EA199800393A priority patent/EA199800393A1/ru
Priority to EP96936906A priority patent/EP0866710A4/fr
Priority to AU74710/96A priority patent/AU706262B2/en
Publication of WO1997015308A1 publication Critical patent/WO1997015308A1/fr
Publication of WO1997015308A9 publication Critical patent/WO1997015308A9/fr
Priority to NO981810A priority patent/NO981810L/no

Links

Classifications

    • 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
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • 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
    • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
    • A61K31/549—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame having two or more nitrogen atoms in the same ring, e.g. hydrochlorothiazide
    • 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
    • A61K31/435—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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease

Definitions

  • the invention relates to compositions and methods for use in limiting undesired bone loss in a vertebrate at risk of such bone loss, in treating conditions that are characterized by undesired bone loss or by the need for bone growth, in treating fractures, and in treating cartilage disorders. More specifically, the invention concerns the use of specific classes of compounds identified or characterized by a high throughput screening assay.
  • transforming growth factor ⁇ the heparin-binding growth factors (acidic and basic fibroblast growth factor), the insulin-like growth factors (insulin-like growth factor I and insulin-like growth factor II), and a recently described family of proteins called bone mo ⁇ hogenetic proteins (BMPs). All of these growth factors have effects on other types of cells, as well as on bone cells.
  • the BMPs are novel factors in the extended transforming growth factor ⁇ superfamily They were first identified by Wozney J et al. Science (1988) 242.1528-34, using gene cloning techniques, following earlier descriptions characterizing the biological activity in extracts of demineralized bone (Urist M Science ( 1965) 150 893-99).
  • BMP2 and BMP4 can induce new bone formation when they are injected locally into the subcutaneous tissues of rats (Wozney J Molec Reprod Dev (1992) 32 160- 67). These factors are expressed by normal osteoblasts as they differentiate, and have been shown to stimulate osteoblast differentiation and bone nodule formation in vitro as well as bone formation in vivo (Harris S et al. J. Bone Miner Res ( 1994) 9 855-63) This latter property suggests potential usefulness as therapeutic agents in diseases which result in bone loss
  • the cells which are responsible for forming bone are osteoblasts.
  • osteoblasts differentiate from precursors to mature bone-forming cells, they express and secrete a number of enzymes and structural proteins ofthe bone matrix, including Type-1 collagen, osteocalcin, osteopontin and alkaline phosphatase (Stein G et al. Curr Opm Cell Biol ( 1990) 2 1018-27, Harris S et al. ( 1994), supra) They also synthesize a number of growth regulatory peptides which are stored in the bone matrix, and are presumably responsible for normal bone formation These growth regulatory peptides include the BMPs (Harris S et al.
  • BMPs 1, 2, 3, 4, and 6 are expressed by cultured cells prior to the formation of mineralized bone nodules (Hams S et al. ( 1994), supra)
  • the BMPs are expressed by cultured osteoblasts as they proliferate and differentiate
  • the BMPs are potent stimulators of bone formation in vitro juv ⁇ in vivo, there are disadvantages to their use as therapeutic agents to enhance bone healing
  • BMPs Receptors for the bone mo ⁇ hogenetic proteins have been identified in many tissues, and the BMPs themselves are expressed in a large variety of tissues in specific temporal and spatial patterns. This suggests that BMPs may have effects on many tissues other than bone, potentially limiting their usefulness as therapeutic agents when administered systemically Moreover, since they are peptides, they would have to be administered by injection These disadvantages impose severe limitations to the development of BMPs as therapeutic agents There is a plethora of conditions which are characterized by the need to enhance bone formation. Perhaps the most obvious is the case of bone fractures, where it would be desirable to stimulate bone growth and to hasten and complete bone repair. Agents that enhance bone formation would also be useful in facial reconstruction procedures.
  • bone deficit conditions include bone segmental defects, periodontal disease, metastatic bone disease, osteolytic bone disease and conditions where connective tissue repair would be beneficial, such as healing or regeneration of cartilage defects or injury.
  • connective tissue repair would be beneficial, such as healing or regeneration of cartilage defects or injury.
  • chronic condition of osteoporosis including age-related osteoporosis and osteoporosis associated with post-menopausal hormone status
  • Other conditions characterized by the need for bone growth include primary and secondary hype ⁇ arathyroidism, disuse osteoporosis, diabetes-related osteoporosis, and glucocorticoid-related osteoporosis.
  • the compounds ofthe present invention may modulate metabolism, proliferation and/or differentiation of normal or aberrant cells or tissues. There are currently no satisfactory pharmaceutical approaches to managing any of these conditions.
  • US Patent 5, 280, 040 discloses a class of compounds which are 3, 4-diaryl chromans. These compounds can be considered derivatives of 2,3,4 triphenyl butanol, where the hydroxy at the 1 -position forms an ether with the ortho position ofthe phenyl group substituted at the 4-position ofthe butanol.
  • the parent 3, 4-diaryl chromans do not contain nitrogen atoms in the aromatic moieties or their linkers
  • a preferred compound, centchroman contains a nitrogen substituent only in one ofthe substituents on a phenyl moiety.
  • the present invention discloses compounds useful for limiting or treating bone deficit conditions, and for other uses that should be apparent to those skilled in the art from the teachings herein. Disclosure ofthe Invention
  • the invention provides compounds that can be administered as ordinary pharmaceuticals and have the metabolic effect of enhancing bone growth
  • the compounds ofthe invention can be identified using an assay for their ability to activate control elements associated with these factors
  • the invention is directed to methods and compositions for stimulating the growth of skeletal (bone) tissue, which methods and compositions use, as active ingredients, compounds wherein two aromatic systems are coupled so as to be spaced apart from each other by about 1.5 to about 15 Angstroms
  • the thus-linked systems may include at least one nitrogen atom other than a ring substituent.
  • the compounds useful in the invention can be described as having the formula Ar'-linker-Ar 2 , wherein each of Ar 1 and Ar 2 is independently an aromatic system and the linker portion ofthe formula spaces Ar 1 and Ar 2 apart by a distance of approximately 1.5-15 Angstroms Ar 1 , Ar 2 and the linker may optionally be substituted with non interfering substituents
  • Ar 1 , Ar 2 and the linker may optionally be substituted with non interfering substituents
  • the compounds ofthe invention also contain at least one additional heteroatom selected from the group consisting of N, S and O, independent of any substituent
  • the invention is directed to methods to treat bone disorders using the compounds described and to pharmaceutical compositions for this use
  • Figure 1 shows the dose response curve for the compound, designated 59-0008
  • Figures 2 and 3 show illustrative compounds ofthe invention and the results obtained with them in an in vitro test.
  • the immortalized cells were stably transfected with a plasmid containing a luciferase reporter gene driven by a mouse BMP2 promoter (- 2736/1 14 bp), and responded in a dose-dependent manner to recombinant human BMP2
  • the assay utilizes cells transformed permanently or transiently with constructs in which the promoter of a bone mo ⁇ hogenetic protein, specifically BMP2 or BMP4, is coupled to a reporter gene, typically luciferase
  • a reporter gene typically luciferase
  • BMP promoter-active compounds or “active compounds”
  • BMP promoter-active compounds are useful in promoting bone or cartilage growth, and thus in the treatment of vertebrates in need of bone or cartilage growth BMP promoter-active compounds can be examined in a variety of other assays that test specificity and toxicity
  • non-BMP promoters or response elements can be linked to a reporter gene, typically luciferase
  • bone deficit is meant an imbalance in the ratio of bone formation to bone reso ⁇ tion, such that, if unmodified, the subject will exhibit less bone than desirable, or the subject's bones will be less intact and coherent than desired. Bone deficit may also result from fracture, from surgical intervention or from dental or periodontal disease.
  • cartilage defect is meant damaged cartilage, less cartilage than desired, or cartilage that is less intact and coherent than desired.
  • Representative uses ofthe compounds ofthe present invention include: repair of bone defects and deficiencies, such as those occuring in closed, open and non-union fractures; prophylactic use in closed and open fracture reduction; promotion of bone healing in plastic surgery; stimulation of bone ingrowth into non-cemented prosthetic joints and dental implants; elevation of peak bone mass in pre-menopausal women; treatment of growth deficiencies; treatment of peridontal disease and defects, and other tooth repair processes; increase in bone formation during distraction osteogenesis; and treatment of other skeletal disorders, such as age-related osteoporosis, post-menopausal osteoporosis, glucocorticoid-induced osteoporosis or disuse osteoporosis and arthritis.
  • bone defects and deficiencies such as those occuring in closed, open and non-union fractures
  • prophylactic use in closed and open fracture reduction promotion of bone healing in plastic surgery
  • stimulation of bone ingrowth into non-cemented prosthetic joints and dental implants elevation of peak bone mass in pre-menopausal women
  • treatment of growth deficiencies treatment of
  • the compounds ofthe present invention can also be useful in repair of congenital, trauma-induced or surgical resection of bone (for instance, for cancer treatment), and in cosmetic surgery Further, the compounds ofthe present invention can be used for limiting or treating cartilage defects or disorders, and may be useful in wound healing or tissue repair. Bone or cartilage deficit or defect can be treated in vertebrate subjects by administering compounds ofthe invention which exhibit certain structural and functional characteristics.
  • the compositions ofthe invention may be administered systemically or locally.
  • the compounds herein are formulated for parenteral (e.g., intravenous, subcutaneous, intramuscular, intraperitoneal, intranasal or transdermal) or enteral (e.g., oral or rectal) delivery according to conventional methods.
  • Intravenous administration can be by a series of injections or by continuous infusion over an extended period. Administration by injection or other routes of discretely spaced administration can be performed at intervals ranging from weekly to once to three times daily.
  • the compounds disclosed herein may be administered in a cyclical manner (administration of disclosed compound; followed by no administration; followed by administration of disclosed compound, and the like). Treatment will continue until the desired outcome is achieved.
  • pharmaceutical formulations will include a compound ofthe present invention in combination with a pharmaceutically acceptable vehicle, such as saline, buffered saline, 5% dextrose in water, borate-buffered saline containing trace metals or the like.
  • Formulations may further include one or more excipients, preservatives, solubilizers, buffering agents, albumin to prevent protein loss on vial surfaces, lubricants, fillers, stabilizers, etc.
  • Methods of formulation are well known in the art and are disclosed, for example, in Remington's Pharmaceutical Sciences. Gennaro. ed . Mack Publishing Co., Easton PA 1990, which is inco ⁇ orated herein by reference
  • Pharmaceutical compositions for use within the present invention can be in the form of sterile, non-pyrogenic liquid solutions or suspensions, coated capsules, suppositories, lyophilized powders, transdermal patches or other forms known in the art.
  • Local administration may be by injection at the site of injury or defect, or by insertion or attachment of a solid carrier at the site, or by direct, topical application of a viscous liquid, or the like.
  • the delivery vehicle preferably provides a matrix for the growing bone or cartilage, and more preferably is a vehicle that can be absorbed by the subject without adverse effects. Delivery of compounds herein to wound sites may be enhanced by the use of controlled-release compositions, such as those described in pending U.S. Patent Application No. 07/871,246 (corresponding to WIPO publication WO 93/20859, which is inco ⁇ orated herein by reference in its entirety). Films of this type are particularly useful as coatings for prosthetic devices and surgical implants.
  • the films may, for example, be wrapped around the outer surfaces of surgical screws, rods, pins, plates and the like. Implantable devices of this type are routinely used in orthopedic surgery.
  • the films can also be used to coat bone filling materials, such as hydroxyapatite blocks, demineralized bone matrix plugs, collagen matrices and the like.
  • a film or device as described herein is applied to the bone at the fracture site. Application is generally by implantation into the bone or attachment to the surface using standard surgical procedures.
  • biodegradable films and matrices may include other active or inert components. Of particular interest are those - o -
  • growth factors agents that promote tissue growth or infiltration, such as growth factors.
  • growth factors for this pu ⁇ ose include epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), transforming growth factors (TGFs), parathyroid hormone (PTH), leukemia inhibitory factor (LIF), and insulin-like growth factors (IGFs) and the like
  • EGF epidermal growth factor
  • FGF fibroblast growth factor
  • PDGF platelet-derived growth factor
  • TGFs transforming growth factors
  • PTH parathyroid hormone
  • LIF leukemia inhibitory factor
  • IGFs insulin-like growth factors
  • Biodegradable films or matrices include calcium sulfate, tricalcium phosphate, hydroxyapatite, polylactic acid, polyanhydrides, bone or dermal collagen, pure proteins, extracellular matrix components and the like and combinations thereof. Such biodegradable materials may be used in combination with non-biodegradable materials, to provide desired mechanical, cosmetic or tissue or matrix interface properties
  • Alternative methods for delivery of compounds ofthe present invention include use of ALZET osmotic minipumps (Alza Co ⁇ ., Palo Alto, CA), sustained release matrix materials such as those disclosed in Wang et al. (PCT Publication WO 90/11366), electrically charged dextran beads, as disclosed in Bao et al. (PCT Publication WO 92/03125); collagen-based delivery systems, for example, as disclosed in Ksander et al. Ann. Surg. (1990) 21 1(3).288-94, methylcellulose gel systems, as disclosed in Beck et al. J. Bone Min. Res. (1991) 6( 1 1) 1257-65; and alginate-based systems, as disclosed in
  • the compounds ofthe present invention may also be used in conjunction with agents that inhibit bone reso ⁇ tion Antireso ⁇ tive agents, such as estrogen, bisphosphonates and calcitonin, are preferred for this pu ⁇ ose More specifically, the compounds disclosed herein may be administered for a period of time (for instance, months to years) sufficient to obtain correction of a bone deficit condition. Once the bone deficit condition has been corrected, the vertebrate can be administered an anti-reso ⁇ tive compound to maintain the corrected bone condition. Alternatively, the compounds disclosed herein may be adminstered with an anti-reso ⁇ tive compound in a cyclical manner (administration of disclosed compound, followed by anti-reso ⁇ tive, followed by disclosed compound, and the like).
  • Aqueous suspensions may contain the active ingredient in admixture with pharmacologically acceptable excipients, comprising suspending agents, such as methyl cellulose; and wetting agents, such as lecithin, lysolecithin or long-chain fatty alcohols.
  • the said aqueous suspensions may also contain preservatives, coloring agents, flavoring agents and sweetening agents in accordance with industry standards
  • Preparations for topical and local application comprise aerosol sprays, lotions, gels and ointments in pharmaceutically appropriate vehicles which may comprise lower aliphatic alcohols, polyglycols such as glycerol, polyethylene glycol, esters of fatty acids, oils and fats, and silicones.
  • the preparations may further comprise antioxidants, such as ascorbic acid or tocopherol, and preservatives, such as p-hydroxybenzoic acid esters.
  • Parenteral preparations comprise particularly sterile or sterilized products.
  • Injectable compositions may be provided containing the active compound and any ofthe well known injectable carriers. These may contain salts for regulating the osmotic pressure.
  • the osteogenic agents can be inco ⁇ orated into liposomes by any ofthe reported methods of preparing liposomes for use in treating various pathogenic conditions.
  • the present compositions may utilize the compounds noted above inco ⁇ orated in liposomes in order to direct these compounds to macrophages, monocytes, other cells and tissues and organs which take up the liposomal composition.
  • the liposome-inco ⁇ orated compounds ofthe invention can be utilized by parenteral administration, to allow for the efficacious use of lower doses ofthe compounds.
  • Ligands may also be inco ⁇ orated to further focus the specificity ofthe liposomes.
  • Suitable conventional methods of liposome preparation include, but are not limited to, those disclosed by Bangham, A.D. et al. J Mol Biol (1965) 23:238-252, Olson, F. et al. Biochim Biophys Acta (1979) 557:9-23, Szoka, F. et al. Proc NatlAca Sci USA (1978) 75:4194-4198, Mayhew, E. et al. (1984) 775: 169175, Kim, S. et al. Biochim Biophys Acta (1983) 728:339:348, and Mayer, et al. Biochim Biophys Acta (1986) 858: 161-168.
  • the liposomes may be made from the present compounds in combination with any ofthe conventional synthetic or natural phospholipid liposome materials including phospholipids from natural sources such as egg, plant or animal sources such as phosphatidylcholine, phosphatidylethanolamine. phosphatidylgiycerol, sphingomyelin, phosphatidylserine. or phosphatidylinositol.
  • Synthetic phospholipids that may also be used, include, but are not limited to: dimyristoylphosphatidylcholine, dioleoylphosphatidylcholine, dipalmitoylphosphatidylcholine and distearoylphosphatidycholine, and the corresponding synthetic phosphatidylethanolamines and phosphatidylglycerols. Cholesterol or other sterols, cholesterol hemisuccinate, glycolipids. cerebrosides. fatty acids, gangliosides.
  • sphingolipids l,2-bis(oleoyloxy)-3-(trimethyl ammonio) propane (DOTAP), N-[l-(2,3- dioleoyl) propyl-N,N,N-trimethylammonium chloride (DOTMA), and other cationic lipids may be inco ⁇ orated into the liposomes, as is known to those skilled in the art.
  • the relative amounts of phospholipid and additives used in the liposomes may be varied if desired. The preferred ranges are from about 60 to 90 mole percent ofthe phospholipid; cholesterol, cholesterol hemisuccinate, fatty acids or cationic lipids may be used in amounts ranging from 0 to 50 mole percent.
  • the amounts ofthe present compounds inco ⁇ orated into the lipid layer of liposomes can be varied with the concentration ofthe lipids ranging from about 0.01 to about 50 mole percent.
  • the liposomes with the above formulations may be made still more specific for their intended targets with the inco ⁇ oration of monoclonal antibodies or other ligands specific for a target.
  • monoclonal antibodies to the BMP receptor may be inco ⁇ orated into the liposome by linkage to phosphatidylethanolamine (PE) inco ⁇ orated into the liposome by the method of Leserman, L. et al. Nature (1980) 288:602-604.
  • PE phosphatidylethanolamine
  • Veterinary uses ofthe disclosed compounds are also contemplated. Such uses would include limitation or treatment of bone or cartilage deficits or defects in domestic animals, livestock and thoroughbred horses.
  • the compounds described herein can also modify a target tissue or organ environment, so as to attract bone-forming cells to an environment in need of such cells
  • the compounds ofthe present invention may also be used to stimulate growth of bone-forming cells or their precursors, or to induce differentiation of bone-forming cell precursors, either in vitro or ex vivo
  • the term "precursor cell” refers to a cell that is committed to a differentiation pathway, but that generally does not express markers or function as a mature, fully differentiated cell
  • the term "mesenchymal cells” or “mesenchymal stem cells” refers to pluripotent progenitor cells that are capable of dividing many times, and whose progeny will give ⁇ se to skeletal tissues, including cartilage, bone, tendon, ligament, marrow stroma and connective tissue (see A Caplan J. Orthop. Res.
  • osteogenic cells includes osteoblasts and osteoblast precursor cells More particularly, the disclosed compounds are useful for stimulating a cell population containing marrow mesenchymal cells, thereby increasing the number of osteogenic cells in that cell population
  • hematopoietic cells are removed from the cell population, either before or after stimulation with the disclosed compounds
  • osteogenic cells may be expanded
  • the expanded osteogenic cells can be infused (or reinfused) into a vertebrate subject in need thereof
  • a subject's own mesenchymal stem cells can be exposed to compounds ofthe present invention ex vivo, and the resultant osteogenic cells could be infused or directed to a desired site within the subject, where further proliferation and/or differentiation ofthe osteogenic cells can occur without immunorejection
  • the cell population exposed to the disclosed compounds may be immortalized human fetal osteoblastic or osteogenic cells If such cells are infused or implanted in a vertebrate subject, it may be advantageous to
  • an "effective amount" of a composition is that amount which produces a statistically significant effect
  • an “effective amount” for therapeutic uses is the amount ofthe composition comprising an active compound herein required to provide a clinically significant increase in healing rates in fracture repair, reversal of bone loss in osteoporosis, reversal of cartilage defects or disorders, prevention or delay of onset of osteoporosis, stimulation and/or augmentation of bone formation in fracture non-unions and distraction osteogenesis; increase and/or acceleration of bone growth into prosthetic devices; and repair of dental defects.
  • Such effective amounts will be determined using routine optimization techniques and are dependent on the particular condition to be treated, the condition ofthe patient, the route of administration, the formulation, and the judgment ofthe practitioner and other factors evident to those skilled in the art.
  • the dosage required for the compounds ofthe invention (for example, in osteoporosis where an increase in bone formation is desired) is manifested as a statistically significant difference in bone mass between treatment and control groups. This difference in bone mass may be seen, for example, as a 5-20% or more increase in bone mass in the treatment group.
  • Other measurements of clinically significant increases in healing may include, for example, tests for breaking strength and tension, breaking strength and torsion, 4-point bending, increased connectivity in bone biopsies and other biomechanical tests well known to those skilled in the art.
  • the dosage ofthe compounds ofthe invention will vary according to the extent and severity ofthe need for treatment, the activity ofthe administered compound, the general health ofthe subject, and other considerations well known to the skilled artisan. Generally, they can be administered to a typical human on a daily basis on an oral dose of about 0.1 mg/kg- 1000 mg/kg, and more preferably from about 1 mg/kg to about 200 mg/kg The parenteral dose will appropriately be 20-100% ofthe oral dose.
  • osteogenic activity ofthe compounds used in the methods ofthe invention can be verified using in vitro screening techniques, such as the assessment of transcription of a reporter gene coupled to a bone mo ⁇ hogenetic protein-associated promoter, as described above, or in alternative assays such as the following:
  • This assay is similar to that described by Gowen M. & Mundy G. J Immunol (1986) 136:2478-82. Briefly, four days after birth, the front and parietal bones of ICR Swiss white mouse pups are removed by microdissection and split along the sagittal suture. The bones are incubated in BGJb medium (Irvine Scientific, Santa Ana, CA) plus 0.02% (or lower concentration) ⁇ -methylcyclodextrin, wherein the medium also contains test or control substances, at 37°C in a humidified atmosphere of 5% CO 2 and 95% air for 96 hours
  • auxiliary assays can be used as controls to determine non-BMP promoter- mediated effects of test compounds
  • mitogenic activity can be measured using screening assays featuring a serum-response element (SRE) as a promoter and a luciferase reporter gene.
  • SRE serum-response element
  • these screening assays can detect signalling through SRE-mediated pathways, such as the protein kinase C pathway
  • SRE-mediated pathways such as the protein kinase C pathway
  • an osteoblast activator SRE-luciferase screen and an insulin mimetic SRE-luciferase screen are useful for this pu ⁇ ose
  • test compound stimulation of c AMP response element (CRE)-mediated pathways can also be assayed
  • cells transfected with receptors for PTH and calcitonin two bone-active agents
  • CRE-luciferase screens can detect elevated cAMP levels
  • the BMP promoter specificity of a test compound can be examined through use of these types of auxiliary assays
  • Bone changes are measured on sections cut 200 ⁇ m apart, over 4 adjacent lxl mm fields on both the injected and noninjected sides ofthe calvaria. New bone is identified by its characteristic woven structure, and osteoclasts and osteoblasts are identified by their distinctive mo ⁇ hology. Histomo ⁇ hometry software (OsteoMeasure, Osteometrix, Inc., Atlanta) is used to process digitizer input to determine cell counts and measure areas or perimeters.
  • Lead compounds can be fu ⁇ her tested in intact animals using an in vivo, dosing assay.
  • Prototypical dosing may be accomplished by subcutaneous, intraperitoneal or oral administration, and may be performed by injection, sustained release or other delivery techniques.
  • the time period for administration of test compound may vary (for instance, 28 days as well as 35 days may be appropriate)
  • An exemplary, in vivo subcutaneous dosing assay may be conducted as follows
  • test compound, positive control compound, PBS, or vehicle alone is administered subcutaneously once per day for 35 days All animals are injected with calcein nine days and two days before sacrifice (two injections of calcein administered each designated day). Weekly body weights are determined.
  • the animals are weighed and bled by orbital or cardiac puncture. Serum calcium, phosphate, osteocalcin, and CBCs are determined. Both leg bones (femur and tibia) and lumbar vertebrae are removed, cleaned of adhering soft tissue, and stored in 70% ethanol for evaluation, as performed by peripheral quantitative computed tomography (pQCT; Ferretti, J. Bone (1995) 17:353S-64S), dual energy X-ray abso ⁇ tiometry (DEXA; Laval- Jeantet A et al. Calcif Tissue /wt/ (1995) 56 14-18; J. Casez et al.
  • pQCT peripheral quantitative computed tomography
  • pQCT Ferretti, J. Bone (1995) 17:353S-64S
  • DEXA dual energy X-ray abso ⁇ tiometry
  • test compounds can also be tested in acute ovariectomized animals (prevention model) using an in vivo dosing assay.
  • assays may also include an estrogen-treated group as a control.
  • An exemplary subcutaneous dosing assay is performed as follows In a typical study, 80 three-month-old female Sprague-Dawley rats are weight- matched and divided into eight groups, with ten animals in each group This includes a baseline control group of animals sacrificed at the initiation ofthe study; three control groups (sham ovariectomized (sham OVX) + vehicle only, ovariectomized (OVX) + vehicle only, PBS-treated OVX), and a control OVX group that is administered a compound known to promote bone growth Three dosage levels ofthe compound to be tested are administered to the remaining three groups of OVX animals.
  • test compound positive control compound, PBS, or vehicle alone is administered subcutaneously once per day for 35 days.
  • test compound can be formulated in implantable pellets that are implanted for 35 days, or may be administered orally, such as by gastric gavage
  • All animals including sham OVX/vehicle and OVX vehicle groups, are injected intraperitoneally with calcein nine days and two days before sacrifice (two injections of calcein administered each designated day, to ensure proper labeling of newly formed bone) Weekly body weights are determined At the end ofthe 35-day cycle, the animals' blood and tissues are processed as described above Lead compounds may also be tested in chronic OVX animals (treatment model)
  • An exemplary protocol for treatment of established bone loss in ovariectomized animals that can be used to assess efficacy of anabolic agents may be performed as follows Briefly, 80 to 100 six month old female, Sprague-Dawley rats are subjected to sham surgery (sham OVX) or ovariectomy (OVX) at time 0, and 10 rats are sacrificed to serve as baseline controls. Body weights are recorded weekly during the experiment. After approximately 6 weeks of bone depletion (42 days), 10 sham OVX and 10 OVX rats are randomly selected for sacrifice as depletion period controls. Ofthe remaining animals, 10 sham OVX and 10 OVX rats are used as placebo-treated controls.
  • the remaining OVX animals are treated with 3 to 5 doses of test drug for a period of 5 weeks (35 days).
  • a group of OVX rats can be treated with an agent such as PTH, a known anabolic agent in this model (Kimmel et al. Endocrinology (1993) 132: 1577-84).
  • PTH a known anabolic agent in this model
  • the proximal left and right tibiae are used for pQCT measurements, cancellous bone mineral density (BMD) (gravimetric determination), and histology, while the midshaft of each tibiae is subjected to cortical BMD or histology.
  • BMD cancellous bone mineral density
  • the femurs are prepared for pQCT scanning ofthe midshaft prior to biomechanical testing.
  • LV lumbar vertebrae
  • LV2 are processed for BMD (pQCT may also be performed);
  • LV3 are prepared for undecalcified bone histology; and LV4 are processed for mechanical testing.
  • All ofthe compounds ofthe invention contain two aromatic systems, Ar 1 and Ar 2 , spaced apart by a linker at a distance of 1.5- 15 A, and may contain at least one nitrogen atom. Both the systems represented by Ar 1 and Ar 2 may contain non-interfering substituents.
  • the non-interfering substituents on the aromatic system represented by Ar 1 and the non-interfering substituents on the aromatic system represented by Ar 2 are represented in the formulae herein by R' and R b , respectively; however, it is recognized that the designation of one Ar as Ar 1 and the other as Ar 2 is arbitrary.
  • alkyl preferably lower alkyl 1-4C
  • alkenyl 1-6C, preferably 1-4C
  • alkynyl 1-6C, preferably 1-4C
  • substituents that do not interfere with the beneficial effect ofthe compounds ofthe invention on bone in treated subjects
  • substituents that do not interfere with the beneficial effect ofthe compounds ofthe invention on bone in treated subjects
  • Preferred non-interfering substituents include hydrocarbyl groups of 1-6C, including saturated and unsaturated, linear or branched hydrocarbyl as well as hydrocarbyl groups containing ring systems; halo groups, alkoxy, hydroxy, amino, monoalkyl- and dialkylamino where the alkyl groups are 1-6C, CN, CF 3 , and COOR.
  • R * and R b substituents may typically be 0-4 or 0-5 depending on the available positions in the aromatic system, preferred embodiments include those wherein the number of R * is 0, 1 or 2 and of R b is 0, 1 or 2.
  • the linker group, L may be a covalent bond or any group having a valence of at least two and covering a linear distance of from about 1.5 to about 15 Angstroms, including those that contain cyclic moieties, that meet this spatial requirement.
  • Useful linkers are divided, by definition herein, into three general categories: (1) flexible non- conjugating linkers, (2) flexible conjugating linkers, and (3) constrained linkers. The preferred choice of linker will depend on the choices for Ar 1 and Ar 2 . Not all ofthe linkers defined below are suitable for all Ar 1 and Ar 2 combinations.
  • flexible non-conjugating linkers are those that link only one position of Ar 1 to one position of Ar 2 , and provide only a single covalent bond or a single chain between Ar 1 and Ar 2 .
  • the chain may contain branches, but may not contain ⁇ -bonds (except in the branches) or cyclic portions in the chain.
  • the linker atoms in the chain itself rotate freely around single covalent bonds, and thus the linker has more than two degrees of freedom.
  • Particularly useful flexible non-conjugating linkers are those ofthe formulae: -NR-, -CR 2 -, -S-, or -O-, wherein R is H or alkyl (1-6C), more preferably H or lower alkyl (1-4C) and more preferably H.
  • linker within this group is dependent on the nature of Ar 1 and Ar 2 .
  • Flexible conjugating linkers are those that link only one position of Ar 1 to one position of Ar 2 , but inco ⁇ orate at least one double or triple bond and/or one or more cyclic systems and thus have only two degrees of freedom.
  • a flexible conjugating linker may form a completely conjugated ⁇ -bond linking system between Ar 1 and Ar 2 , thus providing for co-planarity of Ar 1 and Ar 2 .
  • Constrained linkers are those that have more than one point of attachment to either or both Ar 1 and Ar 2 and, thus, generally allow for only one degree of freedom Constrained linkers most frequently form fused 5- or 6-membered cyclic moieties with Ar 1 and/or Ar 2 where either Ar 1 or Ar 2 has at least one substituent appropriately positioned to form a second covalent bond with the linker, e.g., where Ar 2 is a phenyl group with a reactive, ortho-positioned substituent, or is derivatized to the linker directly at the ortho position.
  • Ar 1 is a substituted or unsubstituted aromatic system containing a six-membered heterocycle and the compounds useful in the invention have the formula:
  • R" is a non-interfering substituent
  • m is an integer of 0-4
  • each dotted line represents an optional ⁇ -bond
  • each Z is independently N, NR, O, S, CR or CR 2 , where each R is independently H or alkyl (1-6C);
  • X is O, S, SO or SO 2 ;
  • L is a flexible linker;
  • Ar 2 is a substituted or unsubstituted 6-membered aromatic ring.
  • a particularly preferred set of embodiments is ofthe formula:
  • Ar is a six-membered (un)substituted aromatic ring. Allowable substituents on this aromatic ring include: halogen, straight or branched chain lower alkyl, alkenyl, or alkynyl, optionally substituted by a six-membered aromatic, cyclic alkyl, or cyclic alkenyl ring, hydroxyl, siloxy, acyloxy, straight or branched chain lower alkoxyl, benzoyl, carboalkoxy, carbamoyl optionally substituted at nitrogen by lower chain alkyl or phenyl, or carboxy, in which R 6 is taken from the group: hydrogen, or straight or branched chain lower alkyl; R 2 and R 5 are individually taken from the group: H, hydroxy, siloxy, acyloxy, halo, cyano, straight or branched chain lower alkyl, or straight or branched chain lower alkoxyl;
  • R 3 and R 4 are individually taken from the group: H, halogen, straight or branched chain lower alkyl, alkenyl, or alkynyl optionally substituted by a six-membered aromatic, cyclic alkyl, or cyclic alkenyl ring, hydroxyl, siloxy, acyloxy, straight or branched chain lower alkoxyl, benzoyl, carboalkoxy, carbamoyl optionally substituted at nitrogen by lower chain alkyl or phenyl, and carboxy;
  • X and Y are either: NR 8 and N, respectively, in which case X and Y are singly bonded, or CR 9 and CR 10 , respectively, in which case X and Y are doubly bonded, wherein R 8 is either H or lower alkyl; R 9 and R 10 are individually taken from the group: H, halo, and lower alkyl;
  • Z is taken from the group: O, S, SO, and SO 2 , or salts thereof.
  • R 2 , R 3 , R 4 , R 5 are as defined above and R is taken from the group: Ar,
  • R' is a non-interfering substituent
  • n is an integer of 0 and 5;
  • L is a flexible linker which does not contain nitrogen
  • Ar 2 is a substituted or unsubstituted phenyl or a substituted or unsubstituted naphthyl. Particularly preferred embodiments of this group of compounds are those ofthe formula.
  • R , 35 is taken from the group: H, hydroxy, alkoxyl, acyloxy, and silyloxy;
  • R is either Ar, or COAr, in which Ar is (un)substituted phenyl in which the allowed substituents are taken from the group: H, hydroxy, (un)substituted alkoxy, acyloxy, siloxy, (un)substituted alkyl, (un)substituted alkenyl, or (un)substituted alkynyl, carboxy, carboalkoxy, carbamoyl optionally substituted at nitrogen by lower chain alkyl, and aryl;
  • R 37 is taken from the group: H, hydroxy, alkoxy, halo, acyloxy, and siloxyl;
  • R 38 is taken from the group: H, hydroxy, alkoxy, acyloxy, siloxy, (un)substituted alkoxy, acyloxy, siloxy, (un)substituted alkyl, (un)substituted alkenyl, and (un)substituted alkynyl, or salts thereof.
  • Compounds of general structure XXXV can be prepared by treating an acetophenone of general structure XXXVI with an appropriate aldehyde of general structure XXXVTI under either basic or acidic conditions,
  • R 35 , R 36 , R 37 , and R 38 are as defined above, followed, optionally, by conversion of any one or more ofthe groups R 35 , R 36 , R 37 , and R 38 into new groups R 35 , R 36 , R 37 , and R 38 by deprotection, coupling, addition, substitution, or elimination, and, if desired, by converting a compound ofthe general structure XXXV into its salt or setting it free from its salt.
  • Specific representative compounds of general structure XXXV include: 2,4-dimethoxy-2'-hydroxychalcone 1 -(2-hydroxyphenyl)-3-(4-methoxyphenyl)propan-
  • Still another group of compounds useful in the invention are those ofthe formula:
  • R' is a non-interfering substituent
  • n is an integer of 0 and 5
  • L is a constrained linker
  • Ar 2 is a substituted or unsubstituted phenyi or a substituted or unsubstituted naphthyl.
  • Particularly preferred compounds in this group are those of formulas IX, XIV, and XX as follows: - R 1 1
  • R" and R 12 are individually taken from the group
  • R 13 , R 14 and R 17 are individually taken from the group
  • R 15 is taken from the group Hydroxy, (un)subst ⁇ tuted C ⁇ . )2 alkoxy, C ⁇ . ]2 alkyl, (un)substituted alkenyl, and acetyloxy,
  • R 16 is taken from the group H, hydroxy, (un)substituted lower alkoxy, acetoxy, (un)substituted alkyl, and (un)substituted alkenyl, where R n , R 12 may form a 5-7 member (un)substituted carbocycle or heterocycle, where R 15 , R 16 may form a 5-7 member (un)substituted carbocyclic or heterocyclic ring,
  • X 1 is either carbonyl or CH 2 , and the dotted line may be a double bond, in which permissible substituents on the above mentioned substituted groups include Lower alkyl, lower alkoxyl, hydroxy, siloxy, halo, carboxyl, and aryl, with the following provisions if X 1 is carbonyl and if R 15 is hydroxy and if only one of R u , R 12 , or R 13 is hydroxy, then at least one of R 14 , R 16 , and R 17 must be other than H, or if R 15 is alkoxy, and if R 11 , R 12 , R 13 together are H, then R 17 can be neither H nor hydroxy, or if R 15 is (un)substituted alkoxy, and if R n , R 12 , and R 13 together consist of only H, or H and one or two alkoxy, and R 17 is H, then R 14 must be other than H,
  • R 15 is hydroxy or alkoxy, and if R u , R 12 , R 13 together consist of only H, or H and one or two alkoxy, or H and only one or two alkyl, and R 17 is C ⁇ alkyl, then at least one of R 14 and R 16 must be other than H; or if R 15 is hydroxy and if R 11 , R 12 , R 13 , R 14 , and R 16 all are H, R 17 must be neither H nor hydroxy; or if R 15 is iso-propoxy, and if R 11 , R 12 , and R 13 together consist of only H, or H and one or two hydroxys, then at least one of R 14 , R 16 , R 17 must be other than H; or if R 15 is 1,5 di(lower) alkyl C5.10 alkyl, then at least one of R 1 1 , R 12 , R 13 , R 14 , R 16 , and R 17 must be other than H; or salts thereof.
  • the groups R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , and R 17 are as defined above, followed, optionally, by the conversion of any one or more of groups R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , and R 17 into new groups R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , and R 17 by deprotection, dehydrogenation, addition, substitution, or elimination, and, if desired, by converting a compound ofthe general structure IX into its salt or setting it free from its salt.
  • 1,3,5-trihydroxybenzene is allowed to react with iso-pentynyl chloride. followed by catalytic hydrogenation, to give product 2.
  • the compound 2 is allowed to react with the acid chloride 3 to provide the ketone 4.
  • Ketone 4 is treated with ethyloxalyl chloride in pyridine at 0°C to afford an ester, which is hydrolyzed in aqueous acetone containing sodium carbonate to give the acid 5.
  • acid 5 When heated in refluxing toluene, acid 5 undergoes decarboxylation to give compound 6, which upon treatment with 2,3-dichloro-5,6-dicyano- 1,4-benzoquinone gives the isoflavanoid 7.
  • R 18 and R 19 are individually taken from the group:
  • R , 20 is taken from the group: H, hydroxy, halo, lower chain alkyl, acyloxy, and siloxy; in which R 21 is taken from the group: Alkyl, alkenyl, alkynyl, aralkyl, (un)substituted phenyl, (un)substituted naphthyl, thienyl, furanyl, and pyridyl; and R 22 is comprised of a C 3 -6 carbohydrate moiety; or salts thereof.
  • R 23 , R 24 , R 25 , R 26 are individually taken from the group: H. hydroxy, (un)substituted alkoxy, siloxy, (un)substituted alkyl,
  • Y 1 is taken from the group: O, -OCH 2 CH 2 O-, -OCH 2 CH 2 S-, -OCH 2 CH 2 CH 2 O-, -SCH 2 CH 2 CH 2 S-, and -SCH 2 CH 2 S-;
  • X 2 is taken from the group: CH 2 , O, and S,
  • R 23 and R 26 must be other than H, or salts thereof
  • Compounds ofthe general structure XX can be prepared by reacting amides of general structure XXI with 5ec-butyl lithium and tetramethylethylenediamine in THF, followed by addition of benzaldehydes of general structure XXII, and the addition of acid.
  • the resulting lactones of general structure XXIII can be reduced by catalytic hydrogenation or treatment with activated zinc in acid, followed by dehydration with trifluoracetic anhydride, ._• ,_»,(_- ⁇ --,
  • diaryl ethers of general structure XXIV with sulfuric acid, alumium trichloride, trifluoracetic anhydride, or similar reagent,
  • R , R , R , R are as defined above, followed, optionally, by conversion of any one or more ofthe groups R 23 , R 24 , R 25 , R 26 into new groups R 23 , R 24 , R 25 , R 26 by deprotection, coupling, addition, substitution, or elimination, and, if desired, by converting a compound ofthe general structure XX into its salt or setting it free from its salt.
  • X 3 is NR 27 , X 4 is CR 30 , X 5 is O, X 6 is CR 31 , X 7 is O ' , or X 3 s NR 30 , X 4 s CR 27 or N, X 5 is NR 31 , X 6 is CR 28 , X 7 is O " or S ' or X 3 s NR 27 .
  • X 4 is CR 28 or N
  • X 5 is NR 30
  • X 6 is CR 29
  • X 7 is NR 32
  • X 3 s NR 30 X 4 s CR 27 or N
  • X 5 is NR 28
  • X 6 is CR 29
  • X 7 is NR 32
  • X 4 is CR 30
  • X 5 is S, X 6 s CR 31 , X 7 s NR 32 ; or s NR 30 .
  • X 4 s CR 27 , X 5 is S, X 6 s CR 28 , X 7 s NR 32 , or X 3 s S, X 4 s CR 30 , X NR 27 X 6 s CR 31 , X 7 s O ' or S ' ; or X 3 s S, X 4 s CR 30 , X 5 s NR 27 , X 6 s CR 28 , X 7 s NR 32 ; or X 3 is S, X 4 s CR 27 , X 5 s NR 30 , X 6 s CR 28 , X 7 s NR 32 or X 3 ,s S, X 4 s CR 30 , X 5 s S, X 6 is CR 27 , X 7 is NR 32 ;
  • X 4 is CR 27 or N, X 5 is NR 31 , X 6 is N, X 7 is O " or S-; or X 3 NR 27 .
  • X 4 is CR 30 , X 5 is NR 28 , X 4 is N, X 7 is NR 32 or CZ Z 3 ; or X 3 s NR 27 .
  • X 4 is CR 28 or N, X 5 is NR 30 , X 4 is N, X 7 is NR 32 or
  • X 3 is NR 30 .
  • X 4 is N, X s is S, X 6 is CR 31 , X 7 is O ; or X 3 is S, X 4 is CR 27 , X 5 is NR 30 , X 6 is N, X 7 is NR 32 ; or X 3 is S, X 4 is CR 30 , X 5 is NR 27 , X 6 is N, X 7 is NR 32 ; or X 3 is O or S, X 4 is N, X 5 is NR 30 , X 6 is N, X 7 is NR 32 ; in which
  • R 27 , R 28 and R 29 are individually straight or branched chain lower alkyl;
  • R 30 and R 31 are individually taken from the group: hydrogen, straight or branched chain (un)substituted alkyl, (un)substituted aromatic, in which the substituents may include Halogen, straight or branched chain lower alkyl, alkenyl, alkynyl optionally substituted by a six-membered aromatic, cyclic alkyl, or cyclic alkenyl ring, hydroxyl, straight or branched chain alkoxyl, benzoyl, carboalkoxy, carbamoyl optionally substituted at nitrogen by lower chain alkyl or phenyl, or carboxy,
  • R 32 is taken from the group:
  • Ar is a six-membered (un)substituted aromatic ring, in which substituents on this ring may include Halogen, straight or branched chain lower alkyl, alkenyl, alkynyl optionally substituted by a six-membered aromatic, cyclic alkyl. or cyclic alkenyl ring, hydroxyl, straight or branched chain alkoxyl. benzoyl, carboalkoxy, carbamoyl optionally substituted at nitrogen by lower chain alkyl or phenyl, or carboxy, R 33 is taken from the group Hydrogen, and straight or branched chain alkyl,
  • Z 2 and Z 3 are individually taken from the group CN and
  • R 34 is taken from the group Hydrogen, straight or branched chain alkyl, and (un)substituted aromatic, or salts thereof
  • Compounds of general structure XXV above can be prepared by treating compounds of general structure XXVI, where X 8 is NR 30 or S, X 9 is CR 30 or N, X 10 is NR 30 or S, Z 4 is CO 2 H, CO 2 R ' ° or CN, with acid chlorides or anhydrides,
  • X XV I or by reacting compounds of general structure XXVII, where X 11 is NR 30 or S, X 12 is N or CR 30 , X 13 is halogen, SMe, or OEt, with amines, sulfides or enolates.
  • R 22 , R 28 , R 29 , R 3U , R 31 . R 32 , R 33 , and R 34 are as defined above, followed. optionally, by conversion of any one or more of the groups R 22 . R 28 , R 29 , R 30 , R 31 , R 32 , R 33 , and R 34 into new groups R 22 , R 28 , R 29 , R 30 , R 31 , R 32 , R 3 ⁇ and R 34 by deprotection, coupling, addition, substitution, or elimination, and, if desired, by converting a compound ofthe general structure XXV into its salt or setting it free from its salt. Specific representatives of compounds ofthe general structure XXV include:
  • Example 1 Compound 59-0008 was synthesized according to the procedure of McDonald, W S., et al. Chem Comm ( 1969) 392-393; Irving, H. N N. H et al. Anal Chim Acta (1970) 49:261-266. Briefly, 10.0 g of dithizone was taken up in 100 ml EtOH and 50 ml AcOH and heated at reflux for 18 h. After cooling, this was diluted first with 100 ml water and then with 50 ml IN NaOH. This was then further neutralized by the addition of 6 N NaOH to bring the pH to 5.0. This deep purple mixture was then concentrated on a rotavapor to remove organics.
  • the 2T3-BMP-2-LUC cells a stably transformed osteoblast cell line described in Ghosh-Choudhury et al. Endocrinology (1996) 137 331-39, referenced above.
  • the cells were cultured using ⁇ -MEM, 10% FCS with 1% penicillin/streptomycin and 1% glutamine ("plating medium"), and were split 1 5 once per week
  • the cells were resuspended in a plating medium containing 4% FCS, plated in microtiter plates at a concentration of 5 x 10 3 cells (in 50 ⁇ l)/well, and incubated for 24 hours at 37°C in 5% CO 2
  • 50 ⁇ l of the test compound or the control in DMSO was added at 2X concentration to each well, so that the final volume was 100 ⁇ l
  • the final serum concentration was 2% FCS, and the final DMSO concentration was 1%.
  • Compound 59-0008 (10 ⁇ M) was
  • the treated cells were incubated for 24 hours at 37°C and 5% CO 2 The medium was then removed, and the cells were rinsed three times with PBS After removal of excess PBS, 25 ⁇ l of IX cell culture lysing reagent (Promega #E153A) was added to each well and incubated for at least ten minutes Optionally, the plates/samples could be frozen at this point.
  • To each well was added 50 ⁇ l of luciferase substrate (Promega #E152A; 10 ml Promega luciferase assay buffer per 7 mg Promega luciferase assay substrate). Luminescence was measured on an automated 96-well luminometer, and was expressed as either picograms of luciferase activity per well or as picograms of luciferase activity per microgram of protein.
  • compound 59-0008 (3-phenylazo-lH-4, l,2-benzothiadiazine) exhibited a pattern of reactivity, as shown in Figure 1.
  • the activity for compound 59-0008 was maximal at a concentration of approximately 3-10 ⁇ M and, more particularly, at about 3 ⁇ M, and thus provided a response of approximately 175 light emission units.
  • other tested compounds were evaluated at various concentrations, and these results were compared to the results obtained for 59-0008 at 10 ⁇ M (which value was normalized to 100). For instance, any tested compound in Figure 2 and Figure 3 that showed greater activity than 10 ⁇ M of 59-0008 would result in a value over 100.
  • Compounds 59-008, 59-0102 and 50-0197 were assayed for effects on the differentiation of cartilage cells, as compared to the action of recombinant human BMP-2. Briefly, a mouse clonal chondrogenic cell line, TMC-23, was isolated and cloned from costal cartilage of transgenic mice containing the BMP-2 gene control region driving SV- 40 large T-antigen, generated as described in Ghosh-Choudhury et al Endocrinology 137:331-39, 1996.
  • TMC-23 cells were plated in 96 well microtiter plates in DMEM containing 10% FCS at 4 x 10 3 cells/well. Two days after plating, the cells were confluent and the medium was replaced with fresh medium containing 10% FCS and different concentrations of compounds or recombinant BMP-2. After an additional 2 or 5 days incubation, the plates were washed twice with PBS, and then lysing solution (0.05% Triton X-100) was added (100 ⁇ l/well).
  • lysing solution 0.05% Triton X-100
  • the cells were lysed by three freeze-thaw cycles of -70°C (30 min), followed by 37°C (30 min with shaking). Twenty microliters of cell lysates were assayed with 80 ⁇ l of 5 mM p- nitrophenol phosphate in 1.5 M 2-amino-2-methyl-propanol buffer, pH 10.3 (Sigma ALP kit, Sigma Chemical Co., St. Louis, MO) for 10 min at 37°C. The reaction was stopped by the addition of 100 ⁇ l of 0.5 M NaOH. The spectrophotometric absorbance at 405 nm was compared to that of p-nitrophenol standards to estimate ALP activity in the samples. The protein content ofthe cell lysates was determined by the Bio-Rad protein assay kit (Bio ⁇ Rad, Hercules, CA). Specific activity was calculated using these two parameters.

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Epidemiology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Rheumatology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Medicinal Preparation (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

L'invention porte sur des composés à deux systèmes aromatiques, liés par covalence par un liant à un ou plusieurs atomes ou par un liant défini comme incluant per se une liaison covalente espaçant les systèmes aromatiques d'une distance comprise entre 1,5 et 15 Å, et s'avérant efficaces pour le traitement des états liés à un déficit osseux. Lesdits composés s'administrent à des vertébrés, seuls ou associés à des additifs qui favorisent la croissance osseuse ou inhibent la résorption osseuse. On peut en évaluer l'activité avant administration en vérifiant leur capacité à effectuer la transcription d'un gène reporter couplé à un promoteur associé à une protéine morphogénétique osseuse et/ou leur capacité à stimuler la croissance de la voûte crânienne dans des systèmes de modèles animaux.
PCT/US1996/017019 1995-10-23 1996-10-23 Compositions et procedes pour le traitement des deficits osseux WO1997015308A1 (fr)

Priority Applications (7)

Application Number Priority Date Filing Date Title
BR9611210-7A BR9611210A (pt) 1995-10-23 1996-10-23 Composições e processos para tratamento de condições ósseas deficitárias
JP09516761A JP2000513324A (ja) 1995-10-23 1996-10-23 骨欠損状態を処置するための組成物および方法
KR1019980702947A KR19990067010A (ko) 1995-10-23 1996-10-23 골결핍 상태의 치료용 조성물 및 그 치료방법
EA199800393A EA199800393A1 (ru) 1995-10-23 1996-10-23 Композиции и способы их применение для лечения поражения костей
EP96936906A EP0866710A4 (fr) 1995-10-23 1996-10-23 Compositions et procedes pour le traitement des deficits osseux
AU74710/96A AU706262B2 (en) 1995-10-23 1996-10-23 Compositions and methods for treating bone deficit conditions
NO981810A NO981810L (no) 1995-10-23 1998-04-22 Sammensetninger og fremgangsmÕter for behandling av tilstander ved bensvikt

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US583095P 1995-10-23 1995-10-23
US60/005,830 1995-10-23

Publications (2)

Publication Number Publication Date
WO1997015308A1 true WO1997015308A1 (fr) 1997-05-01
WO1997015308A9 WO1997015308A9 (fr) 1997-06-26

Family

ID=21717974

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/017019 WO1997015308A1 (fr) 1995-10-23 1996-10-23 Compositions et procedes pour le traitement des deficits osseux

Country Status (13)

Country Link
EP (1) EP0866710A4 (fr)
JP (1) JP2000513324A (fr)
KR (1) KR19990067010A (fr)
CN (1) CN1201393A (fr)
AU (1) AU706262B2 (fr)
BR (1) BR9611210A (fr)
CA (1) CA2235481A1 (fr)
CZ (1) CZ115398A3 (fr)
EA (1) EA199800393A1 (fr)
HU (1) HUP9802319A3 (fr)
NO (1) NO981810L (fr)
PL (1) PL327617A1 (fr)
WO (1) WO1997015308A1 (fr)

Cited By (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999009986A1 (fr) * 1997-08-22 1999-03-04 Kyowa Hakko Kogyo Co., Ltd. Derives de 4-aminoquinazoline
EP0912549A1 (fr) * 1996-06-20 1999-05-06 The Board Of Regents, The University Of Texas System Composes et procedes d'obtention de preparations pharmacologiquement actives et leurs utilisations
WO1999066060A1 (fr) * 1998-06-18 1999-12-23 Aventis Pharma Ltd. Promoteur du gene humain mp52 et procede l'associant dans la recherche de substances utiles
EP0973513A1 (fr) * 1996-10-23 2000-01-26 zymogenetics, Inc. Compositions et methodes pour traiter les etats associes a un deficit osseux
WO2000027822A2 (fr) * 1998-11-06 2000-05-18 Basf Aktiengesellschaft Derives de pyrazole tricyclique
US6080779A (en) * 1996-12-13 2000-06-27 Osteoscreen, Inc. Compositions and methods for stimulating bone growth
WO2000078351A1 (fr) * 1999-06-18 2000-12-28 Mitsubishi Pharma Corporation Promoteurs de l'osteogenese
WO2001017562A1 (fr) * 1999-09-02 2001-03-15 Yamanouchi Pharmaceutical Co., Ltd. Agents promoteurs de l'osteogenese
FR2806408A1 (fr) * 2000-03-17 2001-09-21 Oreal Composition cosmetique comprenant un derive de furane- naphtoquinone, leur utilisation comme agent colorant et derives
US6376476B1 (en) 1996-12-13 2002-04-23 Zymogenetics Corporation Isoprenoid pathway inhibitors for stimulating bone growth
EP1243583A1 (fr) * 1999-12-28 2002-09-25 Eisai Co., Ltd. Composes heterocylciques contenant des groupes de sulfamide
US6462019B1 (en) 1998-07-10 2002-10-08 Osteoscreen, Inc. Inhibitors of proteasomal activity and production for stimulating bone growth
US6462036B1 (en) 1998-11-06 2002-10-08 Basf Aktiengesellschaft Tricyclic pyrazole derivatives
WO2003082808A1 (fr) * 2002-04-03 2003-10-09 Sumitomo Pharmaceuticals Company, Limited. Derives de benzamide
US6649631B1 (en) 1997-10-23 2003-11-18 The Board Of Regents Of The University Of Texas System Compositions and methods for treating bone deficit conditions
US6656904B2 (en) 1998-07-10 2003-12-02 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating bone and hair growth
US6660737B2 (en) 2001-05-04 2003-12-09 The Procter & Gamble Company Medicinal uses of hydrazones
WO2004054978A1 (fr) * 2002-12-16 2004-07-01 Bf Research Institute, Inc. Derive de quinoline utilise comme sonde pour le diagnostic d'une maladie associee a une accumulation de la proteine tau
US6838436B1 (en) 1998-07-10 2005-01-04 Osteoscreen Inc. Inhibitors of proteasomal activity for stimulating bone growth
EP1547996A1 (fr) * 2002-08-30 2005-06-29 BF Research Institute, Inc. Sondes de diagnostic et remedes contre des maladies presentant une accumulation de la proteine du prion et methode de marquage
US7112680B2 (en) 2000-06-05 2006-09-26 Austria Wirtschaftsservice Gesellschaft Mit Beschrankter Haftung Heterocyclic hydrazones for use as anti-cancer agents
EP1815860A2 (fr) 2001-12-26 2007-08-08 Genzyme Corporation Inhibiteurs de transport du phosphate
US7297704B2 (en) 2005-02-17 2007-11-20 Wyeth Cycloalkyfused indole, benzothiophene, benzofuran and idene derivatives
JP2010132699A (ja) * 1998-03-19 2010-06-17 Bionorica Ag 向子宮性作用を持たない、エストロゲン型の器官選択性薬剤としての、アヤメ科及びシミシフガ・ラセモサ（Ｃｉｍｉｃｉｆｕｇａｒａｃｅｍｏｓａ）の抽出物、並びにテクトリゲニンの使用
WO2010088408A2 (fr) * 2009-01-28 2010-08-05 Emory University Antagonistes du récepteur nmda, sélectifs pour des sous-unités, et destinés au traitement d'états neurologiques
US8114995B2 (en) 2008-06-26 2012-02-14 Resverlogix Corp. Methods of preparing quinazolinone derivatives
US8309598B2 (en) 2007-12-19 2012-11-13 Givaudan S.A. Cooling compounds
US8394828B2 (en) 2003-02-03 2013-03-12 Janssen Pharmaceutica, Nv Quinoline-derived amide modulators of vanilloid VR1 receptor
WO2014012889A1 (fr) 2012-07-18 2014-01-23 University College Dublin - National University Of Ireland, Dublin Composés antiangiogéniques
US8889698B2 (en) 2007-02-01 2014-11-18 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US8952021B2 (en) 2009-01-08 2015-02-10 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular disease
US9073878B2 (en) 2012-11-21 2015-07-07 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9212149B2 (en) 2004-04-30 2015-12-15 Takeda Pharmaceutical Company Limited Substituted 2-amidoquinazol-4-ones as matrix metalloproteinase-13 inhibitors
US9238640B2 (en) 2009-03-18 2016-01-19 Resverlogix Corp. Anti-inflammatory agents
US9271978B2 (en) 2012-12-21 2016-03-01 Zenith Epigenetics Corp. Heterocyclic compounds as bromodomain inhibitors
US9610251B2 (en) 2011-11-01 2017-04-04 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
US9757368B2 (en) 2009-04-22 2017-09-12 Resverlogix Corp. Anti-inflammatory agents
US9765039B2 (en) 2012-11-21 2017-09-19 Zenith Epigenetics Ltd. Biaryl derivatives as bromodomain inhibitors
US10111885B2 (en) 2015-03-13 2018-10-30 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102942515A (zh) * 2012-10-22 2013-02-27 暨南大学 一种乙烯桥连吲哚化合物及其合成方法和用途
CN107019687B (zh) * 2017-05-08 2020-10-27 上海市伤骨科研究所 N-(4-氯苯基)-3-羟基-2-萘甲酰胺类化合物的用途
CN111116552B (zh) * 2020-01-17 2022-10-11 河北科技大学 一种喹唑啉酮类化合物及其制备方法
CN114469863B (zh) * 2021-11-26 2023-09-26 南方医科大学南方医院 甾醇脂质体作为牙髓和牙本质药物传递系统的应用
CN114288292B (zh) * 2021-12-10 2023-06-13 中南大学湘雅医院 芳香族化合物作为神经丛素蛋白-b2的激活剂以及在制备治疗骨质疏松症药物中的应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5280040A (en) * 1993-03-11 1994-01-18 Zymogenetics, Inc. Methods for reducing bone loss using centchroman derivatives

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4889851A (en) * 1986-11-21 1989-12-26 Fujisawa Pharmaceutical Co, Ltd. Benzothiadiazine compounds, and pharmaceutical composition comprising the same
JP2724396B2 (ja) * 1987-12-18 1998-03-09 武田薬品工業株式会社 骨粗鬆症予防治療剤
JPH05508386A (ja) * 1989-02-10 1993-11-25 ワシントン　リサーチ　ファンデション 免疫調整剤
JPH06192272A (ja) * 1992-12-24 1994-07-12 Japan Tobacco Inc 新規なトリアゾロベンゾチアジアジン及びトリアゾロベンゾチアジアゼピン誘導体
EP0716086A1 (fr) * 1994-12-09 1996-06-12 Boehringer Mannheim Gmbh Inhibiteurs de métalloprotéinases matricielles à base d'acide malonique
WO1998017267A1 (fr) * 1996-10-23 1998-04-30 Zymogenetics, Inc. Compositions et methodes pour traiter les etats associes a un deficit osseux
WO1998025460A1 (fr) * 1996-12-13 1998-06-18 Zymogenetics, Inc. Compositions visant a stimuler la croissance osseuse et methodes afferentes

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5280040A (en) * 1993-03-11 1994-01-18 Zymogenetics, Inc. Methods for reducing bone loss using centchroman derivatives

Cited By (68)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0912549A1 (fr) * 1996-06-20 1999-05-06 The Board Of Regents, The University Of Texas System Composes et procedes d'obtention de preparations pharmacologiquement actives et leurs utilisations
US6720344B2 (en) 1996-06-20 2004-04-13 The Board Of Regents Of The University Of Texas System Methods and compositions for stimulating osteoblast proliferation or treating malignant cell proliferation and methods for selecting osteoblast proliferation stimulants
EP0912549A4 (fr) * 1996-06-20 2002-01-02 Univ Texas Composes et procedes d'obtention de preparations pharmacologiquement actives et leurs utilisations
EP0973513A1 (fr) * 1996-10-23 2000-01-26 zymogenetics, Inc. Compositions et methodes pour traiter les etats associes a un deficit osseux
EP0973513A4 (fr) * 1996-10-23 2003-03-19 Zymogenetics Inc Compositions et methodes pour traiter les etats associes a un deficit osseux
US6376476B1 (en) 1996-12-13 2002-04-23 Zymogenetics Corporation Isoprenoid pathway inhibitors for stimulating bone growth
US6080779A (en) * 1996-12-13 2000-06-27 Osteoscreen, Inc. Compositions and methods for stimulating bone growth
US7101907B2 (en) 1996-12-13 2006-09-05 Zymogenetics Corporation Topical administration of statins for treatment of bone disorders
WO1999009986A1 (fr) * 1997-08-22 1999-03-04 Kyowa Hakko Kogyo Co., Ltd. Derives de 4-aminoquinazoline
US6649631B1 (en) 1997-10-23 2003-11-18 The Board Of Regents Of The University Of Texas System Compositions and methods for treating bone deficit conditions
JP2010132699A (ja) * 1998-03-19 2010-06-17 Bionorica Ag 向子宮性作用を持たない、エストロゲン型の器官選択性薬剤としての、アヤメ科及びシミシフガ・ラセモサ（Ｃｉｍｉｃｉｆｕｇａｒａｃｅｍｏｓａ）の抽出物、並びにテクトリゲニンの使用
WO1999066060A1 (fr) * 1998-06-18 1999-12-23 Aventis Pharma Ltd. Promoteur du gene humain mp52 et procede l'associant dans la recherche de substances utiles
US7175994B2 (en) 1998-07-10 2007-02-13 Osteoscreen Ip, Llc Inhibitors of proteasomal activity for stimulating hair growth
US6656904B2 (en) 1998-07-10 2003-12-02 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating bone and hair growth
US6462019B1 (en) 1998-07-10 2002-10-08 Osteoscreen, Inc. Inhibitors of proteasomal activity and production for stimulating bone growth
US6958220B2 (en) 1998-07-10 2005-10-25 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating hair growth
US6902721B1 (en) 1998-07-10 2005-06-07 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating bone growth
US6884769B1 (en) 1998-07-10 2005-04-26 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating hair growth
US6838252B2 (en) 1998-07-10 2005-01-04 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating hair growth
US6838436B1 (en) 1998-07-10 2005-01-04 Osteoscreen Inc. Inhibitors of proteasomal activity for stimulating bone growth
US7223554B2 (en) 1998-07-10 2007-05-29 Osteoscreen, Ltd. Inhibitors of proteasomal activity for stimulating hair growth
AU762992B2 (en) * 1998-11-06 2003-07-10 Abbott Gmbh & Co. Kg Tricyclic pyrazole derivatives
WO2000027822A3 (fr) * 1998-11-06 2000-08-10 Basf Ag Derives de pyrazole tricyclique
WO2000027822A2 (fr) * 1998-11-06 2000-05-18 Basf Aktiengesellschaft Derives de pyrazole tricyclique
US6462036B1 (en) 1998-11-06 2002-10-08 Basf Aktiengesellschaft Tricyclic pyrazole derivatives
WO2000078351A1 (fr) * 1999-06-18 2000-12-28 Mitsubishi Pharma Corporation Promoteurs de l'osteogenese
WO2001017562A1 (fr) * 1999-09-02 2001-03-15 Yamanouchi Pharmaceutical Co., Ltd. Agents promoteurs de l'osteogenese
US6787534B2 (en) 1999-12-28 2004-09-07 Eisai Co., Ltd. Sulfonamide-containing heterocyclic compounds
EP1243583A4 (fr) * 1999-12-28 2003-08-20 Eisai Co Ltd Composes heterocylciques contenant des groupes de sulfamide
EP1243583A1 (fr) * 1999-12-28 2002-09-25 Eisai Co., Ltd. Composes heterocylciques contenant des groupes de sulfamide
FR2806408A1 (fr) * 2000-03-17 2001-09-21 Oreal Composition cosmetique comprenant un derive de furane- naphtoquinone, leur utilisation comme agent colorant et derives
EP1134217A3 (fr) * 2000-03-17 2001-11-28 L'oreal Composition cosmétique comprenant un dérivé de furane-naphtoquinone, leur utilisation comme agent colorant, et dérivés.
US7112680B2 (en) 2000-06-05 2006-09-26 Austria Wirtschaftsservice Gesellschaft Mit Beschrankter Haftung Heterocyclic hydrazones for use as anti-cancer agents
US6660737B2 (en) 2001-05-04 2003-12-09 The Procter & Gamble Company Medicinal uses of hydrazones
EP1815860A3 (fr) * 2001-12-26 2007-11-21 Genzyme Corporation Inhibiteurs de transport du phosphate
EP1815860A2 (fr) 2001-12-26 2007-08-08 Genzyme Corporation Inhibiteurs de transport du phosphate
WO2003082808A1 (fr) * 2002-04-03 2003-10-09 Sumitomo Pharmaceuticals Company, Limited. Derives de benzamide
EP1547996A4 (fr) * 2002-08-30 2006-08-02 Bf Res Inst Inc Sondes de diagnostic et remedes contre des maladies presentant une accumulation de la proteine du prion et methode de marquage
EP1547996A1 (fr) * 2002-08-30 2005-06-29 BF Research Institute, Inc. Sondes de diagnostic et remedes contre des maladies presentant une accumulation de la proteine du prion et methode de marquage
WO2004054978A1 (fr) * 2002-12-16 2004-07-01 Bf Research Institute, Inc. Derive de quinoline utilise comme sonde pour le diagnostic d'une maladie associee a une accumulation de la proteine tau
US7118730B2 (en) 2002-12-16 2006-10-10 Bf Research Institute, Inc. Quinoline derivative as diagnostic probe for disease with tau protein accumulation
US8394828B2 (en) 2003-02-03 2013-03-12 Janssen Pharmaceutica, Nv Quinoline-derived amide modulators of vanilloid VR1 receptor
US9475822B2 (en) 2004-04-30 2016-10-25 Takeda Pharmaceutical Company Limited Substituted 2- amidoquinazol-4-ones as matrix metalloproteinase-13 inhibitors
US9212149B2 (en) 2004-04-30 2015-12-15 Takeda Pharmaceutical Company Limited Substituted 2-amidoquinazol-4-ones as matrix metalloproteinase-13 inhibitors
US7297704B2 (en) 2005-02-17 2007-11-20 Wyeth Cycloalkyfused indole, benzothiophene, benzofuran and idene derivatives
US10532054B2 (en) 2007-02-01 2020-01-14 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US9199990B2 (en) 2007-02-01 2015-12-01 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US8889698B2 (en) 2007-02-01 2014-11-18 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular diseases
US8309598B2 (en) 2007-12-19 2012-11-13 Givaudan S.A. Cooling compounds
US8114995B2 (en) 2008-06-26 2012-02-14 Resverlogix Corp. Methods of preparing quinazolinone derivatives
US8952021B2 (en) 2009-01-08 2015-02-10 Resverlogix Corp. Compounds for the prevention and treatment of cardiovascular disease
WO2010088408A2 (fr) * 2009-01-28 2010-08-05 Emory University Antagonistes du récepteur nmda, sélectifs pour des sous-unités, et destinés au traitement d'états neurologiques
WO2010088408A3 (fr) * 2009-01-28 2011-03-31 Emory University Antagonistes du récepteur nmda, sélectifs pour des sous-unités, et destinés au traitement d'états neurologiques
US10131640B2 (en) 2009-03-18 2018-11-20 Resverlogix Corp. Anti-inflammatory agents
US11407719B2 (en) 2009-03-18 2022-08-09 Resverlogix Corp. Anti-inflammatory agents
US9238640B2 (en) 2009-03-18 2016-01-19 Resverlogix Corp. Anti-inflammatory agents
US10882828B2 (en) 2009-03-18 2021-01-05 Resverlogix Corp. Anti-inflammatory agents
US9757368B2 (en) 2009-04-22 2017-09-12 Resverlogix Corp. Anti-inflammatory agents
US9610251B2 (en) 2011-11-01 2017-04-04 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
US10016426B2 (en) 2011-11-01 2018-07-10 Resverlogix Corp. Pharmaceutical compositions for substituted quinazolinones
EP3147280A1 (fr) 2012-07-18 2017-03-29 University College Dublin, National University of Ireland, Dublin Composé anti-angiogène
WO2014012889A1 (fr) 2012-07-18 2014-01-23 University College Dublin - National University Of Ireland, Dublin Composés antiangiogéniques
US9073878B2 (en) 2012-11-21 2015-07-07 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9765039B2 (en) 2012-11-21 2017-09-19 Zenith Epigenetics Ltd. Biaryl derivatives as bromodomain inhibitors
US9278940B2 (en) 2012-11-21 2016-03-08 Zenith Epigenetics Corp. Cyclic amines as bromodomain inhibitors
US9271978B2 (en) 2012-12-21 2016-03-01 Zenith Epigenetics Corp. Heterocyclic compounds as bromodomain inhibitors
US10111885B2 (en) 2015-03-13 2018-10-30 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases
US10772894B2 (en) 2015-03-13 2020-09-15 Resverlogix Corp. Compositions and therapeutic methods for the treatment of complement-associated diseases

Also Published As

Publication number Publication date
EP0866710A1 (fr) 1998-09-30
HUP9802319A2 (hu) 1999-02-01
CA2235481A1 (fr) 1997-05-01
NO981810L (no) 1998-06-22
PL327617A1 (en) 1998-12-21
CN1201393A (zh) 1998-12-09
EA199800393A1 (ru) 1998-12-24
AU706262B2 (en) 1999-06-10
CZ115398A3 (cs) 1998-12-16
AU7471096A (en) 1997-05-15
EP0866710A4 (fr) 2001-07-11
BR9611210A (pt) 1999-12-28
HUP9802319A3 (en) 1999-12-28
NO981810D0 (no) 1998-04-22
KR19990067010A (ko) 1999-08-16
JP2000513324A (ja) 2000-10-10

Similar Documents

Publication Publication Date Title
US5922753A (en) Methods for treating bone deficit conditions with benzothiazole
US6153631A (en) Compositions and methods for treating bone deficit conditions
AU706262B2 (en) Compositions and methods for treating bone deficit conditions
US5919808A (en) Compositions and methods for treating bone deficit conditions
US5948776A (en) Compositions and methods for treating bone deficit conditions
US5965573A (en) Compositions and methods for treating bone deficit conditions
US5990169A (en) Compositions and methods for treating bone deficit conditions
US6342514B1 (en) Compositions and methods for treating bone deficit conditions
US5994358A (en) Compositions and methods for treating bone deficit conditions
US6413998B1 (en) Compositions and methods for treating bone deficit conditions
WO1997015308A9 (fr) Compositions et procedes pour le traitement des deficits osseux
US6251901B1 (en) Compositions and methods for treating bone deficit conditions
US6080779A (en) Compositions and methods for stimulating bone growth
EP0944312B9 (fr) Compositions visant a stimuler la croissance osseuse et methodes afferentes
US6649631B1 (en) Compositions and methods for treating bone deficit conditions
US6642216B2 (en) Isoprenoid pathway inhibitors for stimulating bone growth
WO1998025460A9 (fr) Compositions visant a stimuler la croissance osseuse et methodes afferentes
WO1998017267A9 (fr) Compositions et methodes pour traiter les etats associes a un deficit osseux
EP0973513A1 (fr) Compositions et methodes pour traiter les etats associes a un deficit osseux
AU750816B2 (en) Piperazine derivatives for treating bone deficit conditions
US6017940A (en) Compositions and methods for treating bone deficit conditions
US5939444A (en) Compositions and methods for treating bone deficit conditions
EP1609469A2 (fr) Compositions et méthodes pour stimuler la croissance osseuse
MXPA98003131A (en) Compositions and methods for the treatment of deficitary conditions of hue

Legal Events

Date Code Title Description
WWE Wipo information: entry into national phase

Ref document number: 96197827.9

Country of ref document: CN

AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AU BA BB BG BR CA CN CU CZ EE FI GE HU IL IS JP KG KP KR LC LK LR LT LV MD MG MK MN MX NO NZ PL RO SG SI SK TR TT UA UZ VN AM AZ BY KG KZ MD RU TJ TM

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): KE LS MW SD SZ UG AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN ML MR NE SN TD TG

COP Corrected version of pamphlet

Free format text: PAGES 1/50-50/50,DRAWINGS,REPLACED BY NEW PAGES BEARING THE SAME NUMBER;DUE TO LATE TRANSMITTAL BY THE RECEIVING OFFICE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: PV1998-1153

Country of ref document: CZ

ENP Entry into the national phase

Ref document number: 2235481

Country of ref document: CA

Ref document number: 2235481

Country of ref document: CA

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: PA/a/1998/003131

Country of ref document: MX

WWE Wipo information: entry into national phase

Ref document number: 1019980702947

Country of ref document: KR

WWE Wipo information: entry into national phase

Ref document number: 1996936906

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 199800393

Country of ref document: EA

WWP Wipo information: published in national office

Ref document number: 1996936906

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: PV1998-1153

Country of ref document: CZ

WWP Wipo information: published in national office

Ref document number: 1019980702947

Country of ref document: KR

WWR Wipo information: refused in national office

Ref document number: PV1998-1153

Country of ref document: CZ

WWW Wipo information: withdrawn in national office

Ref document number: 1019980702947

Country of ref document: KR

WWW Wipo information: withdrawn in national office

Ref document number: 1996936906

Country of ref document: EP