[go: up one dir, main page]

WO1997015308A9 - Compositions et procedes pour le traitement des deficits osseux - Google Patents

Compositions et procedes pour le traitement des deficits osseux

Info

Publication number
WO1997015308A9
WO1997015308A9 PCT/US1996/017019 US9617019W WO9715308A9 WO 1997015308 A9 WO1997015308 A9 WO 1997015308A9 US 9617019 W US9617019 W US 9617019W WO 9715308 A9 WO9715308 A9 WO 9715308A9
Authority
WO
WIPO (PCT)
Prior art keywords
bone
substituted
compounds
composition
compound
Prior art date
Application number
PCT/US1996/017019
Other languages
English (en)
Other versions
WO1997015308A1 (fr
Filing date
Publication date
Application filed filed Critical
Priority to KR1019980702947A priority Critical patent/KR19990067010A/ko
Priority to JP09516761A priority patent/JP2000513324A/ja
Priority to EP96936906A priority patent/EP0866710A4/fr
Priority to AU74710/96A priority patent/AU706262B2/en
Priority to BR9611210-7A priority patent/BR9611210A/pt
Priority to EA199800393A priority patent/EA199800393A1/ru
Publication of WO1997015308A1 publication Critical patent/WO1997015308A1/fr
Publication of WO1997015308A9 publication Critical patent/WO1997015308A9/fr
Priority to NO981810A priority patent/NO981810L/no

Links

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.
  • Bone is not a static tissue. It is subject to constant breakdown and resynthesis in a complex process mediated by osteoblasts, which produce new bone, and osteoclasts, which destroy bone. The activities of these cells are regulated by a large number of cytokines and growth factors, many of which have now been identified and cloned. Mundy has described the current knowledge related to these factors (Mundy, G. R. Clin Orthop 324 : 24-28, 1996; Mundy, G. R. J Bone Miner Res 8: S505-10, 1993).
  • 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 morphogenetic 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).
  • Recombinant BMP2 and BMP4 can induce new bone formation when they are injected locally into the subcutaneous tissues of rats (Wozney J. Molet 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 of the bone matrix, including Type-1 collagen, osteocalcin, osteopontin and alkaline phosphatase (Stein G. et al. Curr Opin 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. (1994), supra).
  • BMPs 1, 2, 3, 4, and 6 are expressed by cultured cells prior to the formation of mineralized bone nodules (Harris S. et al. ( 1994), supra). Like alkaline phosphatase, osteocalcin and osteopontin, the BMPs are expressed by cultured osteoblasts as they proliferate and differentiate.
  • BMPs are potent stimulators of bone formation in vitro and in vivo, there are disadvantages to their use as therapeutic agents to enhance bone healing.
  • BMPs Receptors for the bone morphogenetic 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. Also of great significance is the 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
  • Bone fractures are still treated exclusively using casts, braces, anchoring devices and other strictly mechanical means. Further bone deterioration associated with post-menopausal osteoporosis has been decreased or prevented with estrogens or bisphosphonates.
  • 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 of the phenyl group substituted at the 4-position of the 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 of the 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 of the Invention
  • the invention provides compounds that can be administered as ordinary pharmaceuticals and have the metabolic effect of enhancing bone growth.
  • the compounds of the 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 1 -linker-Ar 2 , wherein each of Ar 1 and Ar 2 is independently an aromatic system and the linker portion of the formula spaces Ar 1 and Ar 2 apart by a distance of
  • Ar 1 , Ar 2 and the linker may optionally be substituted with non interfering substituents.
  • the compounds of the 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 of the invention and the results obtained with them in an in vitro test.
  • a rapid throughput screening test for compounds capable of stimulating expression of a reporter gene linked to a BMP promoter is described in U. S. Application Serial No. 08/458,434, filed 2 June 1995, the entire contents of which are incorporated herein by reference.
  • This assay is also described as a portion of a study of immortalized murine osteoblasts (derived from a mouse expressing a transgene composed of a BMP2 promoter driving expression of T-antigen) in Ghosh-Choudhery, N. et al. Endocrinology (1996) 137:331-39.
  • 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 morphogenetic protein, specifically BMP2 or BMP4, is coupled to a reporter gene, typically luciferase. These transformed cells are then evaluated for the production of the reporter gene product; compounds that activate the BMP promoter will drive production of the reporter protein, which can be readily assayed. Over 40,000 compounds have been subjected to this rapid screening technique, and only a very small percentage are able to elicit a level of production of luciferase 5-fold greater than that produced by vehicle. Compounds that activate the BMP promoter share certain structural characteristics not present in inactive compounds.
  • the active compounds (“BMP promoter-active compounds” or “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 and inserted into an appropriate host cell. Cytotoxicity can be determined by visual or microscopic examination of BMP promoter- and/or non-BMP promoter-reporter gene-containing cells, for instance. Alternatively, nucleic acid and/or protein synthesis by the cells can be monitored.
  • tissues may be removed and examined visually or microscopically, and optionally examined in conjunction with dyes or stains that facilitate histologic examination. In assessing in vivo assay results, it may also be useful to examine biodistribution of the test compound, using conventional medicinal chemistry/animal model techniques.
  • limit or “limiting” and “treat” or “treatment” are interchangeable terms.
  • the terms include a postponement of development of bone deficit symptoms and/or a reduction in the severity of such symptoms that will or are expected to develop.
  • the terms further include ameliorating existing bone or cartilage deficit symptoms, preventing additional symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, preventing or reversing bone resorption and/or encouraging bone growth.
  • the terms denote that a beneficial result has been conferred on a vertebrate subject with a cartilage, bone or skeletal deficit, or with the potential to develop such deficit.
  • bone deficit is meant an imbalance in the ratio of bone formation to bone resorption, 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 of the compounds of the 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 peri
  • the compounds of the 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 of the present invention can be used for limiting or treating cartilage defects or disorders, and may be useful in wound healing or tissue repair.
  • 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 of the 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.
  • 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.
  • 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.
  • the biodegradable films and matrices may include other active or inert components.
  • agents that promote tissue growth or infiltration such as growth factors.
  • growth factors include epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), transforming growth factors (TGFs), parathyroid hormone (PTH), leukemia inhibitory factor (LEF), and insulin-like growth factors (IGFs) and the like.
  • growth factors include epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), transforming growth factors (TGFs), parathyroid hormone (PTH), leukemia inhibitory factor (LEF), 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
  • LEF 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 of the present invention include use of ALZET osmotic minipumps (Alza Corp., Palo Alto, CA); sustained release matrix materials such as those disclosed in Wang et al. (PCT Publication WO 90/1 1366);
  • 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.
  • suspending agents such as methyl cellulose
  • 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.
  • Parenteral preparations comprise particularly sterile or sterilized products
  • Injectable compositions may be provided containing the active compound and any of the well known injectable carriers. These may contain salts for regulating the osmotic pressure.
  • the osteogenic agents can be incorporated into liposomes by any of the reported methods of preparing liposomes for use in treating various pathogenic conditions.
  • the present compositions may utilize the compounds noted above incorporated 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-incorporated compounds of the invention can be utilized by parenteral administration, to allow for the efficacious use of lower doses of the compounds.
  • Ligands may also be incorporated to further focus the specificity of the 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 Natl Acad 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 of the conventional synthetic or natural phospholipid liposome materials including phospholipids from natural sources such as egg, plant or animal sources such as
  • Synthetic phospholipids that may also be used, include, but are not limited to dimyristoylphosphatidylcho line, dioleoylphosphatidylcholine, dipalmitoylphosphatidylcholine and distearoylphosphatidycholine, and the corresponding synthetic phosphatidylethanolamines and phosphatidylglycerols. Cholesterol or other sterols, cholesterol hemisuccinate, glycolipids, cerebrosides, fatty acids, gangliosides.
  • sphingolipids 1,2-bis(oleoyloxy)-3-(trimethyl ammonio) propane (DOTAP), N-[1-(2,3-dioleoyl) propyl-N,N,N-trimethylammonium chloride (DOTMA), and other cationic lipids may be incorporated 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 of the phospholipid; cholesterol, cholesterol hemisuccinate, fatty acids or cationic lipids may be used in amounts ranging from 0 to 50 mole percent.
  • the amounts of the present compounds incorporated into the hpid layer of liposomes can be varied with the concentration of the 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 incorporation of monoclonal antibodies or other ligands specific for a target.
  • monoclonal antibodies to the BMP receptor may be
  • PE phosphatidylethanolamine
  • Veterinary uses of the 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.
  • meenchymal cells or “mesenchymal stem cells” refers to pluripotent progenitor cells that are capable of dividing many times, and whose progeny will give rise to skeletal tissues, including cartilage, bone, tendon, ligament, marrow stroma and connective tissue (see A. Caplan J. Orthop. Res. ( 1991 ) 9: 641-50).
  • osteoogenic 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 In a preferred method, 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 of the 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 of the 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 of the 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 of the patient, the route of administration, the formulation, and the judgment of the practitioner and other factors evident to those skilled in the art.
  • the dosage required for the compounds of the 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.
  • General guidance for treatment regimens is obtained from experiments carried out in animal models of the disease of interest.
  • the bones are removed from the incubation media and fixed in 10% buffered formalin for 24-48 hours, decalcified in 14% EDTA for 1 week, processed through graded alcohols, and embedded in paraffin wax .
  • Three ⁇ m sections of the calvaria are prepared. Representative sections are selected for histomorphometric assessment of bone formation and bone resorption. Bone changes are measured on sections cut 200 ⁇ m apart. Osteoblasts and osteoclasts are identified by their distinctive morphology.
  • 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. More specifically, these screening assays can detect signalling through SRE-mediated pathways, such as the protein kinase C pathway. For instance, an osteoblast activator SRE-luciferase screen and an insulin mimetic SRE-luciferase screen are useful for this purpose.
  • test compound stimulation of cAMP response element (CRE)-mediated pathways can also be assayed.
  • CRE cAMP response element
  • cells transfected with receptors for PTH and calcitonin can be used in CRE-luciferase screens to detect elevated cAMP levels.
  • the BMP promoter specificity of a test compound can be examined through use of these types of auxiliary assays.
  • Lead compounds can be further 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:
  • 70 three-month-old female Sprague-Dawley rats are weight-matched and divided into seven groups, with ten animals in each group. This includes a baseline control group of animals sacrificed at the initiation of the study; a control group administered vehicle only, a PBS-treated control group, and a positive control group administered a compound (non-protein or protein) known to promote bone growth. Three dosage levels of the compound to be tested are administered to the remaining three groups.
  • 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. At the end of the 35-day cycle, 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.
  • pQCT peripheral quantitative computed tomography
  • DEXA dual energy X-ray absorptiometry
  • Laval-Jeantet A. et al. Calcif Tissue Intl (1995) 56: 14-18 J. Casez et al. Bone and Mineral (1994) 26: 61- 68
  • histomorphometry J. Casez et al. Bone and Mineral (1994) 26: 61- 68
  • Lead compounds can also be tested in acute ovariectomized animals (prevention model) using an in vivo dosing assay.
  • Such assays may also include an estrogen-treated group as a control.
  • An exemplary subcutaneous dosing assay is performed as follows:
  • 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 of the 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 of the 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 of the 35-day cycle, the animals' blood and tissues are processed as described above.
  • 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. Of the 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 of the 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 of the compounds of the invention contain two aromatic systems, Ar 1 and Ar 2 , spaced apart by a linker at a distance of 1.5-15 ⁇ , 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 a 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. For ease of reference, each is designated separately, it will, however, be evident that the linkers described below, unless palindromic, could thus exist in the compounds in "reverse" order of atoms.
  • the non-interfering substituents can be of wide variety.
  • substituents that do not interfere with the beneficial effect of the compounds of the invention on bone in treated subjects are included alkyl (1-6C, preferably lower alkyl 1-4C), including straight or branched-chain forms thereof, alkenyl (1-6C, preferably 1-4C), alkynyl (1-6C, preferably 1-4C), all of which can be straight or branched chains and may contain further substituents; halogens, including F, Cl, Br and I; siloxy, OR, SR, NR 2 , OOCR, COOR, NCOR,
  • NCOOR and benzoyl, CF 3 , OCF 3 , SCF 3 , N(CF 3 ) 2 , CN, SO, SO 2 R and SO 3 R wherein R is alkyl (1-6C) or is H Where two R a or two R b substituents are in adjacent positions in the aromatic system, they may form a ring. Further, rings may be included in substituents which contain sufficient carbon atoms and heteroatoms to provide this possibility.
  • 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 a 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 a 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 of the 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 of the 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.
  • Flexible conjugating linkers are those that link only one position of Ar 1 to one position of Ar 2 , but incorp 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:
  • n 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.
  • 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,
  • R 3 and R 4 are individually taken from the group: H,
  • R 8 is either H or lower alkyl
  • Z is taken from the group O, S, SO, and SO 2 ,
  • R a 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
  • R 35 is taken from the group: H, hydroxy, alkoxyl, acyloxy, and silyloxy;
  • R 36 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;
  • 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
  • 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,
  • Compounds of general structure XXXV can be prepared by treating an acetophenone of general structure XXXVI with an appropriate aldehyde of general structure XXXVII under either basic or acidic conditions,
  • n is an integer of 0 and 5 ;
  • L is a constrained linker
  • Particularly preferred compounds in this group are those of formulas IX, XIV, and XX as follows:
  • R 11 and R 12 are individually taken from the group :
  • R 13 , R 14 and R 17 are individually taken from the group:
  • R 16 is taken from the group H, hydroxy, (un)substituted lower alkoxy, acetoxy, (un)substituted alkyl, and (un)substituted alkenyl ;
  • R 11 , 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;
  • R 15 is hydroxy and if only one of R 11 , R 12 , or R 13 is hydroxy, then at least one of R 14 , R 16 , and R 17 must be other than H;
  • 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 21 is taken from the group Alkyl, alkenyl,
  • R 22 is comprised of a C 3-6 carbohydrate moiety
  • acids of the general structure XVIII with polyphosphoric acid, trifluoracetic anhydride, or similar reagent,
  • R 23 , R 24 , R 25 , R 26 are individually taken from the group:
  • Y 1 is taken from the group O, -OCH 2 CH 2 O-, -OCH 2 CH 2 S-,
  • diaryl ethers of general structure XXIV with sulfuric acid, alumium trichloride, trifluoracetic anhydride, or similar reagent,
  • X 3 is S
  • X 4 is CR 30
  • X 5 is NR 27
  • X 6 is CR 31
  • X 3 is S
  • X 4 is CR 30
  • X 5 is NR 27
  • X 6 is CR 28
  • X 7 is NR 32 ;
  • X 3 is S
  • X 4 is CR 30
  • X 5 is S
  • X 6 is CR 27
  • X 7 is NR 32 ;
  • X 3 is S, X 4 is CR 30 , X 5 is NR 27 , X 6 is N, X 7 is NR 32 ;
  • 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
  • Z 2 and Z 3 are individually taken from the group: CN and
  • 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 30 or CN, with acid chlorides or anhydrides,
  • XXVII 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 30 , 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 33 , and R 34 by deprotection, coupling, addition, substitution, or elimination, and, if desired, by converting a compound of the general structure XXV into its salt or setting it free from its salt.
  • 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, was employed.
  • the cells were cultured using ⁇ -MEM, 10% FCS with 1%
  • 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.
  • 25 ⁇ l of 1X cell culture lysing reagent (Promega #E153A) was added to each well and incubated for at least ten minutes.
  • 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).

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
KR1019980702947A KR19990067010A (ko) 1995-10-23 1996-10-23 골결핍 상태의 치료용 조성물 및 그 치료방법
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
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
BR9611210-7A BR9611210A (pt) 1995-10-23 1996-10-23 Composições e processos para tratamento de condições ósseas deficitárias
EA199800393A EA199800393A1 (ru) 1995-10-23 1996-10-23 Композиции и способы их применение для лечения поражения костей
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 WO1997015308A1 (fr) 1997-05-01
WO1997015308A9 true 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)

Families Citing this family (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2258822A1 (fr) * 1996-06-20 1997-12-24 Sean Kerwin Composes et procedes d'obtention de preparations pharmacologiquement actives et leurs utilisations
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
AU8748798A (en) * 1997-08-22 1999-03-16 Kyowa Hakko Kogyo Co. Ltd. 4-aminoquinazoline derivatives
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
DE19812204A1 (de) * 1998-03-19 1999-11-04 Plantamed Arzneimittel Gmbh Verwendung von Extrakten aus Cimicifuga racemosa und Belamcanda sinensis als estrogenartiges organselektives Arzneimittel ohne uterotrope Wirkung
JP2000004882A (ja) * 1998-06-18 2000-01-11 Hoechst Marion Roussel Kk ヒトｍｐ５２遺伝子プロモーターおよびこれを用いた有用物質の探索法
US6462019B1 (en) 1998-07-10 2002-10-08 Osteoscreen, Inc. Inhibitors of proteasomal activity and production for stimulating bone growth
US6838436B1 (en) 1998-07-10 2005-01-04 Osteoscreen Inc. Inhibitors of proteasomal activity for stimulating bone growth
US6902721B1 (en) 1998-07-10 2005-06-07 Osteoscreen, Inc. Inhibitors of proteasomal activity for stimulating bone growth
CN1335836A (zh) * 1998-11-06 2002-02-13 巴斯福股份公司 三环吡唑衍生物
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
MXPA02006474A (es) 1999-12-28 2002-11-29 Eisai Co Ltd Compuestos heterociclicos que contienen sulfonamida.
FR2806408B1 (fr) * 2000-03-17 2002-10-11 Oreal Composition cosmetique comprenant un derive de furane- naphtoquinone, leur utilisation comme agent colorant et derives
ATE305466T1 (de) 2000-06-05 2005-10-15 Austria Wirtschaftsserv Gmbh Heterocyclische hydrazone als anti-krebs- wirkstoffe
US6660737B2 (en) 2001-05-04 2003-12-09 The Procter & Gamble Company Medicinal uses of hydrazones
US7119120B2 (en) 2001-12-26 2006-10-10 Genzyme Corporation Phosphate transport inhibitors
US20050182040A1 (en) * 2002-04-03 2005-08-18 Naonori Imazaki Benzamide derivatives
US20050260126A1 (en) * 2002-08-30 2005-11-24 Yukitsuka Kudo Diagnostic probes and remedies for diseases with accumulation of prion protein, and stains for prion protein
EP1574500A1 (fr) * 2002-12-16 2005-09-14 BF Research Institute, Inc. Derive de quinoline utilise comme sonde pour le diagnostic d'une maladie associee a une accumulation de la proteine tau
JP2007527363A (ja) 2003-02-03 2007-09-27 ジヤンセン・フアーマシユーチカ・ナームローゼ・フエンノートシヤツプ バニロイドｖｒ１受容体のモジュレーターであるキノリン誘導アミド
EP1740551B9 (fr) 2004-04-30 2013-01-16 Takeda Pharmaceutical Company Limited Composé amide heterocyclique et utilisation de celui-ci en tant qu'inhibiteur mmp-13
PA8663501A1 (es) 2005-02-17 2006-11-09 Wyeth Wyeth Derivados de indol, benzotiofeno, benzofurano e indeno cicloalquilcondensados
NZ579355A (en) 2007-02-01 2012-03-30 Resverlogix Corp 2-(Aryl)-4-oxo-quinazoline derivatives
WO2009076792A1 (fr) 2007-12-19 2009-06-25 Givaudan Sa Composés rafraîchissants
PL2346837T3 (pl) 2008-06-26 2015-07-31 Resverlogix Corp Sposoby wytwarzania pochodnych chinazolinonu
EP2660238B1 (fr) 2009-01-08 2015-05-06 Resverlogix Corporation Composés pour la prévention et le traitement d'une maladie cardiovasculaire
WO2010088414A2 (fr) * 2009-01-28 2010-08-05 Emory University Potentialisateurs du récepteur nmda, sélectifs pour des sous-unités, et destinés au traitement d'états neurologiques
NZ708314A (en) 2009-03-18 2017-08-25 Resverlogix Corp Quinazolinones for use as anticancer agents
ES2821018T3 (es) 2009-04-22 2021-04-23 Resverlogix Corp Nuevos agentes antiinflamatorios
ES2745471T3 (es) 2011-11-01 2020-03-02 Resverlogix Corp Formulaciones orales de liberación inmediata para quinazolinonas sustituidas
WO2014012889A1 (fr) 2012-07-18 2014-01-23 University College Dublin - National University Of Ireland, Dublin Composés antiangiogéniques
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
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
US9271978B2 (en) 2012-12-21 2016-03-01 Zenith Epigenetics Corp. Heterocyclic compounds as bromodomain inhibitors
WO2016147053A1 (fr) 2015-03-13 2016-09-22 Resverlogix Corp. Compositions et procédés thérapeutiques pour le traitement de maladies associées au complément
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
CN116650495B (zh) * 2021-12-10 2024-04-19 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

Family Cites Families (8)

* 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
US5280040A (en) * 1993-03-11 1994-01-18 Zymogenetics, Inc. Methods for reducing bone loss using centchroman derivatives
EP0716086A1 (fr) * 1994-12-09 1996-06-12 Boehringer Mannheim Gmbh Inhibiteurs de métalloprotéinases matricielles à base d'acide malonique
EP0973513A4 (fr) * 1996-10-23 2003-03-19 Zymogenetics Inc Compositions et methodes pour traiter les etats associes a un deficit osseux
US6022887A (en) * 1996-12-13 2000-02-08 Osteoscreen, Inc. Compositions and methods for stimulating bone growth

Similar Documents

Publication Publication Date Title
AU706262B2 (en) Compositions and methods for treating bone deficit conditions
US5922753A (en) Methods for treating bone deficit conditions with benzothiazole
US5919808A (en) Compositions and methods for treating bone deficit conditions
US6153631A (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
US5948776A (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
WO1997015308A9 (fr) Compositions et procedes pour le traitement des deficits osseux
US6413998B1 (en) Compositions and methods for treating bone deficit conditions
US6251901B1 (en) Compositions and methods for treating bone deficit conditions
US6080779A (en) Compositions and methods for stimulating bone growth
US6649631B1 (en) Compositions and methods for treating bone deficit conditions
EP0944312B9 (fr) Compositions visant a stimuler la croissance osseuse et methodes afferentes
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
US5939444A (en) Compositions and methods for treating bone deficit conditions
US6017940A (en) Compositions and methods for treating bone deficit conditions
EP1609469A2 (fr) Compositions et méthodes pour stimuler la croissance osseuse
HK1085676A (en) Compositions and methods for stimulating bone growth
MXPA98003131A (en) Compositions and methods for the treatment of deficitary conditions of hue