JP2000513324A - Compositions and methods for treating bone defect conditions - Google Patents

Abstract

  (57) [Summary] A compound comprising two aromatic systems covalently linked via a linker containing one or more atoms, or as defined to include itself covalently linked through an aromatic system at a distance of 1.5-15 ° "Linkers" are effective in treating conditions involving bone defects. The compound can be administered to a vertebrate subject alone or in combination with an additional agent that promotes bone growth or inhibits bone resorption. They assess their ability to affect the transcription of a reporter gene linked to a promoter associated with bone morphogenetic proteins and / or stimulate calvarial growth in model animal systems prior to administration Are screened for activity.

Description

DETAILED DESCRIPTION OF THE INVENTION                 Compositions and methods for treating bone defect conditionsTechnical fields   The present invention is directed to the use of undesired bone in vertebrates at risk of such bone loss. Limiting loss, by unwanted bone loss, or the need for bone growth Treating symptoms characterized by gender, treating fractures, and treating Compositions and methods for use in treating bone disorders. More details In particular, the invention has been identified or characterized by high-throughput screening assays. It refers to the use of certain classes of compounds.Background art   Bone is not a static tissue. Bone breaks down bone and osteogenic cells that produce new bone In a complex process mediated by destructive osteoclasts, it constantly collapses and And have been recombined. The activity of these cells depends on many cytokines and proliferation Controlled by factors, many of which are now identified and cloned ing. Mundy describes the current findings on these factors (Mundy, G.R. Clin Orthop 324: 24-28, 1996; Mundy, G.R. J Bone Miner Res 8: S505-10, 1993 ).   There is a great deal of useful information on factors affecting bone destruction and resorption However, information on growth factors that stimulate new bone formation is more limited. Researchers have investigated the source of such activity and found that bone tissue itself It was found to be a reservoir of factors that have the ability to stimulate cells. Therefore, Extracts of bovine bone tissue obtained from slaughterhouses are responsible for maintaining the integrity of the bone structure. Biologically active bone that can stimulate bone cells to proliferate as well as building proteins Also includes growth factors. These latter factors include transforming growth factor β, heparin binding Synthetic growth factors (acidic and basic fibroblast growth factors), insulin-like growth factors (Insulin-like growth factor I and insulin-like growth factor II), and recently Described family of proteins called bone morphogenetic proteins (BMPs) There is. All of these growth factors are effective on other types of cells as well as bone cells You.   BMP is a novel factor in the extended transforming growth factor beta superfamily is there. These were initially obtained using the gene cloning technique, Wozney J. et al., Sci. ence (1988) 242: 1528-34, the biological content of the previously demineralized bone extract Identified following a description of the activity characterization (Urist M. Science (1965) 150: 893-99) Was. Recombinant BMP2 and BMP4 are new when injected locally into rat subcutaneous tissue. Can induce new bone formation (Wozney J. Molec Reprod Dev (1992) 32: 160-67). These factors are expressed when they differentiate by normal osteogenic cells, And differentiation of osteogenic cells and formation of bone nodules in vitro and in vivo (Harris S. et al., J. Bone Miner Res. (1994) 9: 855-63). This latter ability is useful for therapeutics in diseases that cause bone loss. Suggest potential usefulness.   The cells responsible for bone formation are osteogenic cells. Osteogenic cells transform from precursor to mature bone When differentiating into adult cells, these are a number of enzymes and structural proteins of the bone matrix (I Type collagen, osteocalcin, osteopontin, and alkaline phosphine (Including Steatase) and secrete (Stein G. et al., Curr Opin Cell Biol (1990) 2: 1018-27; Harris S. et al., (1994), supra). These are also stored in the bone matrix Synthesizes a number of growth-regulating peptides, and is probably responsible for normal bone formation. These growth regulatory peptides include BMP (Harris S. et al., (1994), supra). The rat's In studies of primary cultures of fetal calvarial osteogenic cells, BMPs 1, 2, 3, 4, and And 6 are expressed from cultured cells prior to the formation of mineralized bone nodules (Harris S. et al.). (1994), supra). Alkaline phosphatase, osteocalcin, and osteocalcin Like opontin, BMPs are proliferated by cultured osteogenic cells, It is expressed when differentiating.   BMP is a strong stimulator of bone formation in vitro and in vivo, but bone recovery There are disadvantages to their use as therapeutic agents to enhance steroids. many Bone morphogenetic protein receptors have been identified in tissues and BMPs themselves. The body is expressed in many types of tissues in a specific time and site manner. this Suggests that BMPs may have effects in many tissues other than bone, If so, it potentially limits its usefulness as a therapeutic. In addition, these Because it is a peptide, it should be administered by infusion. These inconveniences are It places severe restrictions on the development of BMP as a remedy.   Presence of polycythemia, characterized by the need to enhance bone formation . Possibly stimulates bone growth and promotes and completes bone repair The most obvious is the case in which a fracture is desired. Drugs that enhance bone formation are on the face It is also useful in the reconstruction procedure. Other bone defect symptoms include bone segment defects, periodontal disease Recovery, metastatic bone disease, osteolytic bone disease, and cartilage defect or wound Conditions where connective tissue repair is beneficial, such as regeneration. Also age related Osteoporosis, including osteoporosis that develops and osteoporosis associated with postmenopausal hormonal status The chronic symptoms of the disease are also very important. Characterized by need for bone growth Other symptoms include primary and secondary hyperparathyroidism, unused osteoporosis , Diabetes-related osteoporosis, as well as glucocorticoid-related osteoporosis. Additionally or alternatively, the compounds of the present invention may be metabolically, proliferative, and / or normal. Or it can regulate the differentiation of abnormal cells or tissues.   Currently, there are sufficient pharmaceutical approaches to control any of these situations do not do. Fractures are exclusively used for casting bandages, braces, anchors, and other rigorous machinery It is still being treated using conventional means. Additional post-menopausal osteoporosis Bone deterioration is reduced or prevented with estrogen or bisphosphonate Have been.   U.S. Pat. No. 5,280,040 discloses 3,4-diallylchroman, a class of compounds. Disclose. These compounds are considered derivatives of 2,3,4 triphenylbutanol. Wherein the hydroxyl group at the 1-position is a phenyl group substituted at the 4-position of butanol. It forms ether with the ortho position. The parent 3,4-diallylchroman is an aromatic ring The moieties or their linkers do not contain a nitrogen atom. Sene, a preferred compound Tochroman (centchroman) has only one substituent at the phenyl moiety with a nitrogen substituent. Including. These compounds are disclosed in the '040 patent as being useful for treating osteoporosis. ing.   The present invention is directed to limiting or treating the symptoms of bone defects, and from the teachings herein to those skilled in the art. Discloses compounds useful for other uses that should be apparent.Disclosure of the invention   The present invention can be administered as a regular formulation and has metabolic effects that enhance bone growth A compound having the formula: The compounds of the present invention may be involved in the regulation of these factors. Can be identified using assays for their ability to activate the element. Accordingly, the present invention relates to methods and compositions for stimulating the growth of skeletal (bone) tissue. The methods and compositions employ a compound as the active ingredient. Where two Are connected at about 1.5 to about 15 ° from each other. This The linked system (including the linker that connects them) It may contain at least one nitrogen atom.   Thus, compounds useful in the present invention are of the formula Ar¹-Linker-Ar^TwoWhere each Ar¹ And Ar^TwoIs independently an aromatic ring system, and the linker moiety of this formula is Ar¹When Ar^TwoAnd at a distance of about 1.5 to 15 mm. Ar¹, Ar^Two, And The linker can be optionally substituted with a non-interfering substituent. Useful compounds include Independently of all substituents in¹, Ar^TwoAnd / or linker At least one nitrogen atom may be present. Preferably, the compounds of the present invention are also And independently of all substituents, at least one selected from the group consisting of N, S, and O. Both contain one further heteroatom.   Other compounds of the invention contain certain five-membered rings with potential separation.   Accordingly, the present invention provides methods, and methods for treating bone disorders using the described compounds. And pharmaceutical compositions for this use.BRIEF DESCRIPTION OF THE FIGURES   FIG. 1 shows a dose response curve for the compound designated 59-0008.   Figures 2 and 3 show exemplary compounds of the present invention, and The results obtained using them are shown.Embodiment of the Invention   Linked to the BMP promoter, a surrogate for endogenously produced bone morphogenetic product Rapid throughput for compounds that can stimulate the expression of a reporter gene Screening test was conducted in US Ser. No. 08 / 458,4, filed Jul. 2, 1995. 34, the entire contents of which are incorporated herein by reference. This Assay is also described by Ghosh-Choudhery, N. et al. Endcrinology (1996) 137: 331-39. Immortalized mouse osteogenic cells (BMP2 promoter driving T-antigen expression) From mice expressing a transgene consisting of It is. In this study, immortalized cells were derived from the mouse BMP2 promoter (-2736 / 114b Control driven by p) and responding to recombinant human BMP2 in a dose-dependent manner Stable transfection with a plasmid containing the luciferase reporter gene Was turned on.   Briefly, this assay is based on the promoter of the bone morphogenetic protein (specifically, BMP2 or BMP4) is linked to a reporter gene (typically luciferase). ) Utilizing cells that have been persistently or transiently transformed. Next Thus, these transformed cells were evaluated for production of the reporter gene product. Compounds that activate the BMP promoter can be easily assayed. Drives production of the porter protein. More than 40,000 compounds are in this rapid screen Subjected to leaning technology, and only a very small percentage It can induce production levels of luciferase that are 5 times greater than the levels produced. BM Compounds that activate the P promoter are not present in inactive compounds, Share structural characteristics. Active compound ("BMP promoter active compound" or "Active compounds") are useful in promoting bone or cartilage growth, Thus, it is useful for treating vertebrates that require bone or cartilage growth.   BMP promoter active compounds may be used in other assays to test various specificities and toxicity. Can also be tested by Say. For example, a non-BMP promoter or response element The report can be linked to a reporter gene and inserted into a suitable host cell. Cytotoxicity can be, for example, BMP promoter- and / or non-BMP promoter- Determined by visual or microscopic examination of cells containing the reporter gene Can be Alternatively, nucleic acid and / or protein synthesis by cells is monitored. Can be For in vivo assays, tissue is removed and visually or Dyes or dyes that can be examined microscopically and, if necessary, facilitate histological examination It can be tested in combination with a dye. In evaluating the results of in vivo assays Test the biodistribution of the test compound using conventional medicinal chemistry / animal model techniques It can also be useful.   As used herein, “restricted” or “restrict”, and “treatment” "Or" are "interchangeable" terms. This term refers to the occurrence of bone defect symptoms. Has developed or is expected to develop postponement of symptoms and / or symptoms Including reducing the severity of such symptoms. The term further refers to the existing bone Or improving cartilage deficiency symptoms, preventing further symptoms, Ameliorate or prevent a negative cause, prevent or reverse bone resorption, and And / or enhancing bone growth. Therefore, this term is used for cartilage, bone or A spinal motion that has a skeletal defect or has the potential to develop such a defect Means that the subject is given a beneficial result.   By "bone defect" is meant an imbalance in the ratio of bone formation to bone resorption. As a result, if not relieved, the subject will show less bone than desired, or The subject's bone is less intact than desired and is not aggregated. Bone Defects can also be caused by fractures, surgical intervention, or dental or periodontal disease. It may be the result of a disease. By "cartilage defect", damaged cartilage, desired Less cartilage, or less intact than desired, and aggregated Not cartilage is meant.   Representative uses of the compounds of the present invention include: closed fractures, open fractures Bone defects and deficiencies, such as occur in non-union and non-union fractures Repair; prophylactic use in reducing closed and open fractures; for plastic surgery Bone healing in non-cemented artificial joints and bone grafts in bone Stimulation of endoproliferation; increased peak bone density in postmenopausal women; treatment of growth defects; periodontal disease And loss treatment, and other dental restoration procedures; distraction osteo enhanced bone formation during genesis), as well as age-related osteoporosis, postmenopausal bone Osteoporosis, glucocorticoid-induced osteoporosis, or unused osteoporosis and related osteoporosis Treatment of other skeletal disorders, such as arthritis. The compounds of the present invention are congenital, Repair of conductive or surgical removal of bone (eg, for cancer treatment), and beauty It may also be useful in surgery. In addition, the compounds of the invention may be used for cartilage damage or Can be used to limit or treat disorders, and wound healing or tissue repair May be useful for   Bone or cartilage defects or deficiencies can cause certain structural and And can be treated by administration of a compound of the invention that exhibits functional properties. Set of the present invention The composition can be administered systemically or locally. For systemic use, this specification Compounds in parenteral delivery (eg, intravenous, subcutaneous, intramuscular, intraperitoneal, intranasal, Or for transdermal) or for enteral delivery (eg, oral or rectal) Formulated according to conventional methods. Intravenous administration may be by a series of injections or by Obtained by continuous infusion over a prolonged period. Administration by injection or other routes Dosing at other locations is performed at intervals ranging from once a week to three times a day obtain. Alternatively, the compounds disclosed herein may be used in a cyclic manner (disclosed). Compound administration; then no administration; then administration of the disclosed compounds, etc.) obtain. Treatment continues until the desired result is achieved. Generally, pharmaceutical formulations include: Compounds of the present invention can be administered in saline, buffered saline, 5% dextrose in water. Pharmaceutically acceptable vehicles, such as borate buffered saline containing trace elements, etc. Included in combination with vehicle. The formulation may further comprise one or more excipients, preservatives, solubilizers , Buffer, albumin to prevent protein loss on vial surface, lubrication Agents, fillers, stabilizers and the like. The method of formulation is well known in the art, and For example,Remington's Pharmaceutical Sciences, Gennaro, Mack Publish ing Co., Easton PA, 1990, which is incorporated herein by reference. It is shown. Pharmaceutical compositions for use in the present invention are sterile, non-pyrogenic Solutions or suspensions, coated capsules, suppositories, lyophilized powders, It can be a skin patch, or a form known in the art. Topical administration is damaged or missing This can be done by injection at the site of injury, or insertion of a solid carrier into the site. Kuha Attachment can be used or for direct, topical application of viscous liquids, etc. Get more. For topical administration, the delivery vehicle is preferably a bone or cartilage Provides a substrate for growth and, more preferably, is tested without adverse effects. A vehicle that can be absorbed by the body.   Delivery of the compounds herein to the site of injury can be controlled release compositions (e.g., For example, see U.S. Patent Application No. 07 / 871,246 (WIPO Publication No. WO 93/20859). Corresponding compositions, all of which are herein incorporated by reference). Can be enhanced. This type of film is used for artificial devices and external It is particularly useful as a coating for medical implantation. The film is, for example, outside It can be wrapped around an external surface such as a technical screw, rod, pin, plate, or the like. This This type of implantable brace is routinely used in orthopedic surgery. the film Is also a bone filling matrix (hydroxyapatite block, To coat demineralized bone matrix plugs, collagen matrix, etc. Can be used. Generally, the film or prosthesis described herein may be used to Applied to bone in position. Application is generally to the bone using normal surgical procedures. By grafting or mounting to the surface.   In addition to the above copolymers and carriers, biodegradable films and substrates are: It may contain other active or inactive ingredients. The specific purpose is to target a group, such as a growth factor. A reagent that promotes tissue growth or invasion. Exemplary growth factors for this purpose As epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (P DGF), transforming growth factor (TGF), parathyroid hormone (PTH), leukemia inhibitory factor (LIF) , And insulin-like growth factors (IGFs). Drugs that promote bone growth Agents such as bone morphogenetic proteins (US Pat. No. 4,761,471; PCT Publication No. WO 90 / No. 11366), osteogenin (Sampath et al., Proc. Natl. Acad. Sci. . USA (1987) 84: 7109-13), and NaF (Tencer et al., J. Biomed. Mat. Res. (198 9) 23: 571-89) are also preferred. As a biodegradable film or substrate, Calcium phosphate, tricalcium phosphate, hydroxyapa Tight, polybutyric acid, polyanhydride, bone or epidermal collagen, pure protein , Extracellular matrix components, and the like, and combinations thereof. Such biodegradation sex The substance is used in combination with a non-biodegradable substance to provide the desired functional, cosmetic Alternatively, it can provide the properties of a tissue or substrate interface.   Another method for delivery of the compounds of the present invention is the ALZET osmotic minipump (osmo tic minipump) (Alza Corp., Palo Alto, CA); Wang et al. (PCT Publication No. WO 90/11366) Sustained release of substrate material as disclosed in Bao et al. (PCT Publication No. WO 92/03125). Dextran beads having a charge as disclosed in); for example, Ksander et al. (Ann .Surg. (1990) 211 (3): 288-94); a methylcellulose gel system, Be. ck et al. (J. Bone Min. Res. (1991) 6 (11): 1257-65). Delivery system; and disclosed in Edelman et al. (Biomaterials (1991) 12: 619-26). Alginate-based systems, etc. . Another method of maintaining local delivery of bone, known in the art, is imprementation (impre gnate) porous coated metal prosthesis, and incorporated therein Solid plastic rods with the therapeutic compositions to be obtained.   The compounds of the present invention may also be used in combination with an agent that inhibits bone resorption. You. Anti-reabsorption, such as estrogen, bisphosphonates and calcitonin Yields are preferred for this purpose. More specifically, disclosed herein The compound is administered for a period of time sufficient to correct the bone defect symptoms (eg, months to years), Can be administered. Once the bone defect symptoms have been corrected, the vertebrate can To maintain condition, an anti-resorbable compound can be administered. Alternatively, as used herein The disclosed compounds can be used in a cyclical fashion (with the disclosed compounds , Then an anti-resorbent, then a disclosed compound, etc.).   As further formulations, conventional preparations as described below may be used.   Aqueous suspensions contain a pharmaceutically acceptable excipient, such as a suspension of methylcellulose. Agents; and moisturizers (including lecithin, lysolecithin, or long-chain fatty alcohols) )). This aqueous suspension also complies with industry standards. Preservatives, colorings, flavors and even sweeteners.   Topical and topical applications include lower fatty alcohols, polyglycerols such as glycerol Recalls, polyethylene glycols, fatty acid esters, oils and fats, Aerosol spray in a pharmaceutically suitable vehicle which may contain And ointments, gels, and ointments. The preparation may further comprise an antioxidant (eg, as Corbic acid or tocophenol), and preservatives (eg, p-hydrobenze) Acid esters).   Parenteral preparations especially include sterile or sterilized products. Active compounds, and Injectable compositions can be provided that include any of the well-known injectable carriers. this Can contain salts to regulate osmotic pressure. If desired, the osteogenic agent may vary. Any reported preparation of liposomes for use in the treatment of pathogenic conditions Can be incorporated into liposomes by different preparation methods. The composition comprises Macrophages, mononuclear cells, and other cells in which the compound takes up the liposome composition And incorporated into liposomes to target tissues and organs Compounds can be used. The compounds of the present invention incorporated into liposomes are suitable for parenteral administration. Thus, it can be utilized, allowing for efficacious use of smaller doses of the compound. Ligands can also be incorporated to further focus liposome specificity .   Conventional suitable methods for the preparation of liposomes are described in Bangham, A.D. et al., J Mol Biol (196 5) 23: 238-252; Olson, F. et al., Biochem Biophys Acta (1979) 557: 9-23, Szoka. 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. You are not limited to them.   Liposomes can be any conventional synthetic or natural, phospholipid liposome material. (Phospholipids from natural sources, such as egg, plant, or animal sources (eg, Phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycol Serol, sphingomyelin, phosphatidylserine, and phosphatidyl (Including inositol). Can be used Synthetic phospholipids also include, but are not limited to: dimyristoyl Sphatidylcholine, dioleoylphosphatidylcholine, dipalmitoylphos Fatidylcholine, distearoylphosphatidylcholine, and counterparts Synthetic phosphatidylethanolamine and phosphatidylglycerol. Cholesterol or other sterols (cholesterol hemisuccinate, Colipid, cerebroside, fatty acid, ganglioside, sphingolipid, 1, 2-bis (oleoyloxy) -3- (trimethylammonio) propane (DOTAP), N- [ 1- (2,3-dioleoyl) propyl-N, N, N, -trimethylammonium chloride ( DOTMA)), and other cationic lipids are incorporated into liposomes as is known to those skilled in the art. Can be impregnated. The relative amounts of phospholipids and additives used in the liposome Can change if done. The preferred range is about 60-90 mol% phospholipid: cholester Roll, cholesterol hemisuccinate, fatty acids, or cationic lipids It can be used in the range of 0 to 50 mol%. This compound incorporated into the lipid layer of liposomes The amount of matter can vary from about 0.01 to about 50 mol% in lipid concentration.   Using conventional methods, about 20-30% of the compound present in solution is converted into liposomes. Can be trapped: about 70-80% of active compound is wasted. Target When compounds are incorporated into liposomes, virtually all compounds The active compound is incorporated into the system and essentially no active compound is wasted.   Liposomes having the above formulation can be prepared using monoclonal antibody incorporation. More specific with ligands specific for their intended target or other targets Can be made. For example, a monoclonal antibody against the BMP receptor is available from Leserma. n, L. et al. (Nature (1980) 288: 602-604) Incorporated into liposomes by binding to phosphatidylethanolamine (PE) Can be rare.   Veterinary use of the disclosed compounds is also contemplated. These uses include livestock, Limitation of bone or cartilage loss or deficiency in livestock and purebred horses Or treatment. The compounds described herein also cause osteogenic cells to The environment of the target tissue or organ in order to attract Can be modified.   The compounds of the present invention can also be used in vivo, either in vitro or ex vivo. To stimulate the growth of morphogenic cells or their precursors, or the precursors of osteogenic cells It can be used to induce cell differentiation. As used herein, the term "previous `` Competent cells '' are mated as mature, fully differentiated cells that are restricted by the differentiation pathway. Or a cell that does not generally express a function. As used herein, The term “mesenchymal cell” or “mesenchymal stem cell” is a pluripotent progenitor cell that can be divided many times And their offspring are responsible for skeletal tissue (cartilage, bone, tendon, ligament, bone marrow matrix and connective tissue). (See A. Caplan J. Orthop. Res. (1991) 9: 641-50)). . As used herein, the term "osteogenic cells" refers to osteogenic cells and osteoblasts. Contains progenitor cells. More particularly, the disclosed compounds are useful in cells including bone marrow mesenchymal cells Useful for stimulating a group and thereby increasing the number of osteogenic cells in that group It is. In a preferred method, hematopoietic cells are stimulated prior to stimulation with the disclosed compounds. , Or later, is removed from the population of cells. Through the implementation of these methods Thus, the osteogenic cells can swell. Expanded osteogenic cells need these The vertebrate subject can be injected (or re-injected). For example, the mesenchyme of the subject itself Stem cells can be exposed ex vivo to a compound of the present invention, and the resulting bone The progenitor cells can be injected or directly directed to the desired site in the subject. In this case, further proliferation and / or differentiation of osteogenic cells are immune rejection It can happen. Alternatively, a population of cells exposed to the disclosed compounds comprises an immortalized human It can be a fetal osteogenic cell or an osteogenic cell. Such cells are used in vertebrate When implanted or transplanted into a subject, it enhances transplantation and bone or cartilage repair. To "immunoprotect" these non-self cells or to suppress the recipient (Preferably locally) may be beneficial.   In the present invention, an “effective amount” of a composition is an amount that produces a statistically significant effect. . For example, an "effective amount" for therapeutic use includes the active compound herein. The amount of the composition required to provide a clinically significant increase in the treatment rate of: Reversal of bone loss in osteoporosis; reversal of cartilage defect or disorder; osteoporosis Prevention or delayed onset of disease; in unconnected fractures and distraction bone formation Stimulation and / or enhancement of bone formation; increased bone growth and / or prosthetic device Or acceleration; and restoration of dental defects. These effective doses are determined using conventional visualization techniques. Specific condition to be determined and treated, the condition of the patient, the route of administration, the prescription, And the judgment of the practitioner, and other factors apparent to those skilled in the art. Of the present invention The required dosage of the compound (eg, in osteoporosis where increased bone formation is desired) ) Indicates a statistically significant difference in bone mass between the treatment and control groups. Different Is specified as This difference in bone mass is, for example, 5% of the bone mass in the treatment group. It can be considered as an increase of 2020% or more. Other clinically significant increases in treatment Measurements include, for example, breaking strength and tension, breaking strength and torsional force, and four-point bending (4 -point bending), increased connectivity in bone biopsy, and other It may include tests for biomechanical tests well known to those skilled in the art. One to the treatment life rules General guidance comes from experiments performed in animal models of the disease of interest.   The dosage of the compound of the present invention depends on the range and severity of the treatment required, the compound administered. Depending on the activity of the product, the general health of the subject, and other considerations well known to those of skill in the art. I do. In general, these can be about 0.1 mg / kg to 1000 mg / kg daily for a typical human, and More preferably, it can be administered daily on an oral dosage basis of about 1 mg / kg to 200 mg / kg . The parenteral dose is suitably about 20-100% of the oral dose.   Screening assay   The osteogenic activity of the compounds used in the method of the present invention was determined by in vitro screening. Techniques (eg, linked to a bone morphogenetic protein binding promoter, Assessment of reporter gene transcripts), or alternative assays such as: Can be proven:   Newborn mouse calvaria assay technology (in vitro)   This assay is described in Gowen M. & Mundy G. J. Immunol (1986) 136: 2478-82 Similar to the one described. Briefly, ICR Swiss White, 4 days old Microdissection of the frontal and parietal bones of the offspring of a Swiss white mouse It was removed by microdessection and divided along the sagittal suture. Bone on BGJb Ground (Irvine Scientific, Santa Ana, CA) + 0.02% (or low concentration) β-methyl Cyclodextrin (where the medium also contains test or control substances) 5% of CO_TwoAnd incubate for 96 hours at 37 ° C in 95% humidified air. I did it.   Following this, the bone is removed from the incubation medium and 10% buffered Fix with rumarin for 24-48 hours, decalcify with 14% EDTA for 1 week, grade alcohol And embedded in paraffin wax. 3 μm calvarial Sections were prepared. Representative sections were selected for histomorphological evaluation of bone formation and resorption. I chose. Bone changes were measured in sections cut to 200 μm. Osteogenesis Vesicles and osteoclasts were identified by their characteristic morphology.   Other supplemental assays to determine non-BMP promoter-mediated effects of test compounds Can be used as a control for For example, mitogenic activity is Responsive element (SRE) as promoter and luciferase reporter It can be measured using a screening assay characterized by the gene. More specifically , These screening assays are based on SRE-mediated pathways (eg, protein inks). Signal through the Nase C pathway). For example, osteogenic cells -Luciferase screening for inverters and pseudo-insulin SRE-luciferase screening is useful for this purpose. Similarly, cAMP response Stimulation of the element (CRE) -mediated pathway by a test compound can also be assayed. For example, receptors for PTH and calcitonin (two bone-activating factors) The transfected cells are used to detect elevated cAMP levels, It can be used for cellulase screening. Therefore, the BMP promoter characteristics of the test compound The isomerism can be tested by using these complementary assay types.   In vivo assay of the effect of compounds on mouse calvarial bone growth   Four to six weeks old, male ICR Swiss white mice weighing 13-26 gm were administered to each group. Use 4-5 animals per animal. The calvarial bone growth assay is described in PCT application WO 95/24211 (reference and Performed as described in). Briefly, test compounds or The appropriate control vehicle is injected into the subcutaneous tissue through the right calvaria of normal mice I do. Typically, a control vehicle is a vehicle that has solubilized the compound. And PBS containing 5% DMSO or PBS containing Tween (2 μl / 10 ml) There is a vehicle. Animals were sacrificed on day 14 and bone growth was determined by histomorphometry (his tomorphometry). Purification of bone samples from attached tissue for calibration Fix with 10% buffered formalin for 24-48 hours and decalcify with 14% EDTA for 1-3 weeks And treated with grade alcohol; further embedded in paraffin wax . 3-5 μm calvarial sections were prepared and typical sections were taken for bone formation and resorption. Effect Select for histomorphological evaluation of fruit. Sections are placed on a digitizing plate under a microscope Measured using a camera equipped with Lucida to directly trace the image I do. Bone changes on both the injected and uninjected sides of the calvaria Sections of four adjacent 1 × 1 mm sections cut at 200 μm intervals are measured. New bone is identified by its characteristic wavy structure, and osteoclasts and bone shape Adult cells were identified using their unique morphology. Organizational morphometry software (O using steoMeasure, Osteometrix, Inc., Atlanta) Alternatively, digital input processing for determining the perimeter is performed.   Further in vivo assays   Lead compounds are transferred to intact animals using an in vivo dose assay. Can be tested further. Basic dosage is subcutaneous, intraperitoneal, or oral And by injection, sustained release or other delivery techniques. Can be performed. The duration of administration of the test compound can vary (eg, 28 days as well as 35 days). Days may be appropriate). Illustratively, an in vivo subcutaneous dose assay is as follows: Can be implemented in:   In a representative study, 70 3-month-old female Sprague-Dawley (Sprague- Dawley) Rats are combined and divided into 7 groups of 10 rats in each group . This is the baseline control group of animals sacrificed at the start of the study. Control group receiving vehicle only; control group treated with PBS And compounds known to promote bone growth (protein or non- Protein) is administered. To be tested The three dose levels of the compound are administered to the remaining three groups.   Briefly, test compounds, positive control compounds, PBS, or Vehicle only is administered subcutaneously once a day for 35 days. Calcein (c alcein) for 9 days and 2 days before sacrifice (two injections of calcein) Was administered on each specified day). Body weight is measured weekly. At the end of the 35-day cycle, The objects were weighed and bled using an orbital or cardiac puncture. Serum calsi Um, phosphate, osteocalcin, and CBC were determined. Both leg bones (thigh Bone and tibia) and lumbar vertebrae are removed, soft adherent tissue is cleaned and evaluated (peripheral Quantitative computed tomography (pQCT; Ferretti, J. Bone (1995) 17: 353S-64S) , Dual Energy X-ray Absorptiometry (DEXA; Lavel-Jeantet A. et al., Calcif Tiss ue Intl (1995) 56: 14-18; Casez et al., Bone and Mineral (1994) 26: 61-68) And / or performed by tissue morphometry) for storage in 70% ethanol I do. Thus, the effect of a test compound on bone remodeling can be assessed.   The main compound is also expressed in vivo in acute ovariectomized animals (prophylactic model). It can be tested using a dose assay. Such assays are also controlled May include estrogen treatment groups. An exemplary subcutaneous dose assay is described below. Implemented as:   In a representative study, the weight of 80 3-month-old female Sprague-Dawley rats And further divided into 8 groups of 10 animals in each group. This is the start of the study Control group at baseline of animals sacrificed in 3 control groups (Sham oophorectomy (sham OVX) + vehicle only; ovariectomy (OVX) + Vehicle only; PBS treated OVX); and Control OVX group included. The three dose levels of the compound to be tested are Administer to three groups of OVX animals.   Ovariectomy (OVX) induces all OVX animals to induce sham OVX With the items, they were pair-feeded for 35 days of study. Briefly, a trial Test compound, positive control compound, PBS or vehicle only for 1 day It was administered subcutaneously once for 35 days. Alternatively, the test compound is implanted for 35 days, It can be formulated as an implantable pellet or administered orally (eg, by gavage) By gastric gavage). Sham OVX / vehicle and OVX / vehicle Calcein was given intraperitoneally for 9 and 2 days before sacrifice Inject (to ensure proper labeling of the newly formed bone Two injections were given each designated day). Body weight is measured weekly. 35 day cycle Later, the blood and tissues of the animals were processed as described above.   Major compounds can also be tested in chronic OVX animals (treatment model). Anabolic agent Treatment of established bone loss in ovariectomized animals that can be used to evaluate efficiency An exemplary protocol for can be implemented as follows. In short, 80-100 female 6 month old female Sprague-Dawley rats were shampooed at 0 hours. Subject to surgery (sham OVX) or ovariectomy (OVX) and base on 10 rats Slaughter to serve as line control. Weight every week during the experiment Record After almost 6 weeks (42 days) of bone depletion, 10 Sham OVX and 10 OVX rats for slaughter as controls Select randomly. 10 Sham OVX and 10 OVX rats of the remaining animals Is used as a placebo treatment control. Replace the remaining OVX animals with 3 Treatment was for 5 weeks (35 days) at a dose of ５5. OV as a positive control A group of X rats is treated with an agent such as PTH (an anabolic agent known in this model) (Kimmel et al., Endocrinology (1993) 132: 1577-84). Effects on bone formation The following procedure may be followed to determine Femur, tibia, and first to fourth lumbar vertebrae Is excised and collected. PQCT measurement of adjacent left and right tibia, spongy mineral dense (BMD) (weight measurement), and used for histology, while the central diaphysis of each tibia was Provide for MD or histology. Prior to biomechanical examination, the femur should be pQCT Prepared for can. For the lumbar spine (LV), LV2 is BMD (pQCT is also Can be performed); LV3 is prepared for histology of non-decalcified bone; LV4 is then processed for mechanical testing.Properties of compounds useful in the present invention   All of the compounds of the present invention are spaced at a distance of 1.5 ° -15 ° by the linker. Two aromatic Ars separated from each other¹And Ar^TwoAnd at least one It may contain a nitrogen atom. Ar¹And Ar^TwoBoth systems, represented by It may contain non-interfering substituents. Ar¹Aromatic represented by Non-interfering substituents on the system and Ar^TwoThe non-interfering substituent on the aromatic system represented by , Where R^aAnd R^bAnd is represented in the formulas herein. (However While replacing one Ar with Ar¹And the other Ar is Ar^TwoNamed as It is understood that doing so is optional. ) For ease of reference, each , Are named separately. (However, the linker described below has a palindrome structure (palindromic) unless they are present in the compound in the order of the "reverse" atoms. It is clear that you can. In general, non-interfering substituents can be of a wide variety You. Substituents that do not interfere with the beneficial effects of the compounds of the invention on the bone of the subject to be treated Include alkyl (1-6C, preferably lower alkyl 1-4C) (their Including straight or branched chain forms), alkenyl (1-6C, preferably 1-4C), Alkynyl (1-6C, preferably 1-4C), all of which are directly And further substituents (halogen (F, Cl, Br, And I), siloxy, OR, SR, NR_Two, OOCR, COOR, NC OR, NCOOR, and benzoyl, CF_Three, OCF_Three, SCF_Three, N (CF_Three)_Two , CN, SO, SO_TwoR and SO_ThreeR (where R is alkyl (1-6C) Or H). Two R^aOr two R^bSubstituents in aromatic systems When in adjacent positions, they may form a ring. In addition, this possibility is offered. Rings may be included in substituents containing enough carbon and heteroatoms.   Preferred non-interfering substituents include 1-6C hydrocarbyl groups (saturated and unsaturated). Containing linear or branched hydrocarbyl groups and hydrocarbyl groups containing ring systems. ), Halo groups, alkoxy, hydroxy, amino, monoalkyl, and dial Killamino (where the alkyl group includes 1-6C, CN, CF_Three, And COO R).   R^aAnd R^bThe number of substituents will typically be dependent on the available position in the aromatic system. 0-4 or 0-5, and preferred embodiments include R^aIs 0, 1 or 2 and R^bIs 0, 1, or 2 You.   The linker group L has a covalent bond or a valence of at least 2 and It can be any group that includes a straight-line distance between Å and about 15Å (satisfies this spatial requirement (Including those having an annular portion). Useful linkers are defined herein. , Three general categories: (1) flexible non-conjugated linkers; Flexible conjugated linkers, and (3) constrained linkers Divided into The choice of a preferred linker is Ar¹And Ar^TwoDepends on the choice of . All of the linkers defined below are all Ar¹And Ar^TwoCombination of Not always suitable for   As defined herein, a flexible non-conjugated linker is Ar¹of Ar at only one position^TwoAnd one shared connection. Or Ar¹And Ar^TwoOnly a single chain between The chain may include branches, but the chain It may not contain π bonds (except in branches) or rings. Linker source in the chain The linker is larger than 2 because the child itself is free to rotate around a covalent single bond. It has a great degree of freedom. In addition to conjugated bonds, particularly useful flexible non-conjugated phosphorus The car has the formula: -NR-, -CR_Two-, -S-, or -O-. here And R is H or alkyl (1-6C), more preferably H or lower. Alkyl (1-4C), and more preferably H. Equally preferred Has the formula: -NRCO-, -CONR-, -CR_TwoS-, -SCR_Two-, -OCR_Two -, -CR_TwoO-, -NRNR-, -CR_TwoCR_Two-, -NRSO_Two-, -SO_TwoN R-, -CR_TwoCO-, -COCR_Two-, And -NR-NR-CO-CR_Two− Rabi ni its complement-CR_Two—CO—NR—NR— (including its isomeric form) . Also preferred are compounds of the formula: -NR (CR_Two)_TwoNR-, -O (CR_Two)_TwoO- And -S (CR_Two)_TwoS- (including its equidistant body). The resources in this group The best choice of anchor is Ar¹And Ar^TwoDepends on the nature of   The flexible conjugated linker is Ar¹Only one position of Ar^TwoOne place of At least one double or triple bond and / or Or it has only two degrees of freedom because it incorporates one or more ring systems. flexible A suitable conjugated linker is Ar¹And Ar^TwoForms a fully conjugated π-linked link system with Ar¹And Ar^TwoAnd co-planarity is provided. Useful flexible conjugation Examples of linkers include -RC = CR-, -N = N-, -C≡C-, -RC = N-, -N = CR-, -NR-N = CR-, -NR-NR-CO-CR = CR-, etc. Wherein R is H or alkyl (1-6C), preferably H Or lower alkyl (1-4C), and more preferably H.   The tethered linker is Ar¹And Ar^TwoMore than one to either or both Since it has additional points, it generally has only one degree of freedom. Bound The linker is Ar¹And / or Ar^TwoFused 5-membered ring moiety or 6-membered ring having Form the parts most often. Where Ar¹Or Ar^TwoIs either a linker At least one substituent appropriately positioned to form a second covalent bond (Eg, where Ar^TwoIs a phenyl having a reactive ortho-position substituent. Or directly derivatized at the ortho position to a linker). (like this In such cases, the aromatic moiety should be properly called phenylene or naphthylene In general, however, the term "phenyl" or "naphthyl" refers to the monovalent of these moieties. It is used herein to include both divalent forms. ) Especially useful bound Examples of linkers include:Wherein X is O, N, S, or CR, and Y is C R_TwoOr, C = O.   Many of the compounds useful in the present invention are commercially available and are known in the art. It can be synthesized by technical methods. New compounds, useful in the present invention These compounds can likewise be obtained by methods generally known in the art. Wear.   In one set of compounds of the invention, Ar¹Is a substituted or unsubstituted 6-membered heterocyclic ring Compounds that are substituted aromatics and that are useful in the present invention have the formula: Has,   Where R^aIs a non-interfering substituent;   m is an integer from 0 to 4;   Each dashed line represents an optional π-bond;   Each Z is independently N, NR, O, S, CR, or CR_TwoAnd     Wherein each R is independently H or alkyl (1-6C);   X is O, S, SO, or SO_TwoIs;   L is a flexible linker; and   Ar^TwoIs a substituted or unsubstituted 6-membered aromatic ring.   A particularly preferred set of embodiments has the following formula:   Of   here,   R¹Is a group: N = NAr, NR⁶COAr, CONR⁶Ar, CH_TwoOAr, CH_Two NR⁶Selected from Ar, wherein Ar is a 6-membered (un) substituted aromatic ring . Possible substituents on this aromatic ring include:   Halogen, straight or branched lower alkyl, alkenyl, or alkynyl (Optionally substituted by a 6-membered aromatic, cyclic alkyl, or cyclic alkenyl ring ), Hydroxyl, siloxy, acyloxy, linear or branched lower alkyl With coxyl, benzoyl, carboalkoxy, lower alkyl or phenyl Carbamoyl optionally substituted at the nitrogen, or carboxy. ,here,   R⁶Is selected from the group: hydrogen or linear or branched lower alkyl;   R^TwoAnd R^FiveIndependently represent a group: H, hydroxy, siloxy, acyloxy, ha B, cyano, straight or branched lower alkyl, or straight or branched lower alkyl Selected from lower alkoxyl,   R^ThreeAnd R^FourIs independently a group: H, halogen, linear or branched lower alkyl. Kill, alkenyl, or alkynyl (6-membered aromatic, cyclic alkyl, or Optionally substituted by a cyclic alkenyl ring), hydroxyl, siloxy, acyl Roxy, linear or branched lower alkoxyl, benzoyl, carboalkoxy Optionally substituted at the nitrogen by lower-chain alkyl or phenyl , And carboxy;   X and Y are each: NR⁸And N, where X and Y is a single bond) or CR⁹And CR^Ten(In this case X And Y are double-bonded), where   R⁸Is either H or lower alkyl;   R⁹And R^TenIs independently selected from the group: H, halo, and lower alkyl And   Z is a group: O, S, SO, and SO_TwoOr a salt thereof. The compounds of general structure I above can be prepared in a variety of ways, for example:   a) thiohydrazide of general structure II, or the corresponding thiohydrazone, Treating in hot acetic acid with   b) reacting a compound of general structure III with bromine;   c) heating the compound of general structure IV in a protic solvent;   d) reacting a compound of general structure V or VI with sodium hydride;  e) reacting the compound of general structure VII with a base;   f) reacting the pyrylium compound of general structure VIII with a suitable nucleophile;   Where R^Two, R^Three, R^Four, R^FiveIs as defined above, and R is a group: Ar, NHAr, NHNHAr, COAr, carboalkoxy, alcohol Kishi, NR⁶COAr, CH_TwoOAr, and CH_TwoNR⁶Selected from Ar, here And Ar and R⁶Is as described above and, if necessary, then deprotected By coupling, addition, substitution or elimination, or by sulfoxides of sulfur Or one or more groups R, R by oxidation to a sulfone^Two, R^Three, R^Four, R^FiveTo New groups R, R^Two, R^Three, R^Four, R^FiveAnd, if desired, the conversion of general formula I The compound is converted to the salt or it is released from the salt.   Example:   The diphenylthiohydrazone is heated in refluxing acetic acid in air for 30-90 minutes, Obtain Nzochi Asiazen 1.  Certain representative examples of compounds of general structure I include:     3-phenylazo-1H-4,1,2-benzothiazidine     2-phenylazo-2H-benzopyran.   Another group of compounds suitable for use in the methods of the invention are compounds of the formula:   And   Where R^aIs a non-interfering substituent;   n is an integer from 0 to 5;   L is a nitrogen-free flexible linker; and   Ar^TwoIs substituted or unsubstituted phenyl or substituted or unsubstituted naphthyl It is.   A particularly preferred embodiment of this group of compounds has the formula:   Of   here,   R³⁵Represents the group: H, hydroxy, alkoxyl, acyloxy, and silyloxy Selected from   R³⁶Is either Ar or COAr, where Ar is (un) substituted Phenyl (possible substituents here are in the group: H, hydroxy, (un) substituted alkoxy , Acyloxy, siloxy, (un) substituted alkyl, (un) substituted alkenyl, or (Un) substituted alkynyl), carboxy, carboalkoxy, lower chain alkynyl Selected from carbamoyl and aryl optionally substituted at the nitrogen by Re;   R³⁷Represents the group: H, hydroxy, alkoxy, halo, acyloxy, and siloxy Selected from sill;   R³⁸Represents the group: H, hydroxy, alkoxy, acyloxy, siloxy, (non) Substituted alkoxy, acyloxy, siloxy, (un) substituted alkyl, (un) substituted alk And alkynyl and (un) substituted alkynyl, or salts thereof.   Compounds of general structure XXXV can be prepared under general or acidic conditions Treatment of acetophenone of structure XXXVI with the appropriate aldehyde of general structure XXXVII Depending on the process   Alternatively, in the presence of a suitable catalyst such as aluminum trichloride, Treating the appropriate alkyne of XVIII with an acid halide of the general structure XXXIX Therefore,   Alternatively, the general structure XX in the presence of a suitable catalyst (eg, a palladium catalyst) The acid chloride of XIX is converted to (E) -1,2-bis (tri-n-butylstannyl (butyls tanyl)) vinyl stana with ethylene or of general structure XL ne)  Alternatively, the step of treating acetophenone having the general structure XLI with a strong base is carried out. Can be prepared by   Where R³⁵, R³⁶, R³⁷, R³⁸Is defined above and is required Depending on, then, by deprotection, coupling, addition, substitution, or elimination, One or more groups R³⁵, R³⁶, R³⁷, R³⁷, And R³⁸With the new radical R³⁵, R³⁶, R³⁷, And R³⁸To a compound of general structure XXXV, if desired. Convert to a salt or free it from the salt.   Certain representative compounds of general structure XXXV include:   2,4-dimethoxy-2'-hydroxychalcone   1- (2-hydroxyphenyl) -3- (4-methoxyphenyl) propane- 1,3-dione   1,4-dioxo-1,4-diphenylbut-2-ene Is mentioned.   Yet another group of compounds useful in the present invention has the following formula:   Of   Where R^aIs a non-interfering substituent;   n is an integer from 0 and 5;   L is a tethered linker; and   Ar^TwoIs substituted or unsubstituted phenyl or substituted or unsubstituted naphthyl It is.   Particularly preferred compounds in this group are those of formulas IX, XIV and XX as shown below. Is:   here:   R¹¹And R¹²The groups independently:     H, hydroxy, C_1-6Alkoxy, acetyloxy, and C_1-12(Non) Selected from substituted alkyl;   R¹³, R¹⁴, And R¹⁷The groups independently:     H, hydroxy, C_1-6Linear or branched alkoxy, and acetyloxy Selected from   R¹⁵Represents the group: hydroxy, (un) substituted C_1-12Alkoxy, C_1-12Alkyl, ( Selected from un) substituted alkenyl, and acetyloxy;   R¹⁶Represents the group: H, hydroxy, (un) substituted lower alkoxy, acetoxy, (non ) Selected from substituted alkyl, and (un) substituted alkenyl;   Where R¹¹, R¹²Forms a 5- to 7-membered (un) substituted carbocyclic or heterocyclic ring Gain;   Where R¹⁵, R¹⁶Represents a 5- to 7-membered (un) substituted carbocyclic or heterocyclic ring May form;     X¹Is carbonyl or CH_TwoOne of the following:   And the dotted line can be a double bond,   Here, possible substituents of the group of substituents described above include lower alkyl, lower alkyl, Lower alkoxyl, hydroxy, siloxy, halo, carboxyl, and aryl Or salts thereof, with the following conditions:   X¹Is carbonyl, and   R¹⁵Is hydroxy, and if R¹¹, R¹²Or R¹³Only one of the If it is droxy, R¹⁴, R¹⁶, And R¹⁷At least one of which is not H Must be   Alternatively, R¹⁵Is alkoxy, and R¹¹, R¹², R¹³Are both H If R¹⁷Cannot be H or hydroxy,   Alternatively, R¹⁵Is (un) substituted alkoxy, and R¹¹, R¹², And R¹³ Are both H only, or H and 1 or 2 alkoxy, and R¹⁷ Is H¹⁴Must be other than H, Me, or hydroxymethyl Not;   Alternatively, R¹⁵Is hydroxy or alkoxy, and R¹¹, R¹², R¹³Are both H only, or H and 1 or 2 alkoxy, or H and only one or two alkyls, and R¹⁷Is C_1-4If it is alkyl , R¹⁴And R¹⁶At least one of must be other than H;   Alternatively, R¹⁵Is hydroxy and R¹¹, R¹², R¹³, R¹⁴,and R¹⁶Are all H, then R¹⁷Should not be H or hydroxy,   Alternatively, R¹⁵Is isopropoxy and R¹¹, R¹², And R¹³Is When only H or H and one or two hydroxys,¹⁴, R¹⁶, R¹⁷At least one of must be other than H;   Alternatively, R¹⁵Is 1,5 di (lower) alkyl C_5-10When it is alkyl, R¹¹ , R¹², R¹³, R¹⁴, R¹⁶, And R¹⁷At least one of must be other than H Must.   Compounds of the general structure shown above are ketones of structure (X) shown below:Is reacted with an alkyl orthoformate in the presence of a base, Reaction with ethyl oxalyl chloride in the presence of gin, followed by hydrolysis and And decarboxylation method, or   A method of reacting with an alkyl formate in the presence of an alkali metal, or   Reacting with N, N-dialkylformamide in the presence of phosphorus oxychloride The law, or   By reacting with cyanide in the presence of hydrogen halide,   Alternatively, 2-hydroxyisoflavanoid having the following general structure (XI) is dehydrated. Depending on the process:   Or by subjecting a compound of the following general structure XII to catalytic hydrogenation:   Or the corresponding phenylacetate is And then (PF₆)_TwoRh (EtC_FiveMe_Four) (C₆H₆) To reduce A compound of general structure XIII obtained byBy processing   Where the group R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, And R¹⁷Is defined above Followed by deprotection, dehydrogenation, addition, substitution, or By elimination, any one or more groups R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, And And R¹⁷To a new group R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, And R¹⁷Convert to Then, if desired, the compound of general structure IX is converted to a salt thereof or From its salt.Example   1,3,5-Trihydroxybenzene is reacted with isopentynyl chloride. And then catalytic hydrogenation to give product 2. Compound 2 is reacted with acid chloride 3 In response, ketone 4 is obtained. Ketone 4 is converted to ethyl oxane in pyridine at 0 ° C. Treat with salyl chloride, ester (this is an aqueous solution containing sodium carbonate) Hydrolysis in acetone to give acid 5). Heated in refluxing toluene In this case, acid 5 undergoes decarboxylation to give compound 6, which is converted to 2,3-dichloromethane. By treating with loro-5,6-dicyano-1,4-benzoquinone, Obtain ravanoid 7.   Particular representative examples of compounds of general structure IX include:   Tectorigenin   Robustone   Robuston methyl ether   7,2 ', 4'-trihydroxyisoflavone   6,2 ', 3'-trihydroxy-7,4'-dimethoxyisoflavan   8,4'-dimethoxy-7-hydroxyisoflavone   The compound of XIV has the following structure: Or a salt thereof,   here,   R¹⁸And R¹⁹The groups independently:     H, hydroxy, (un) substituted alkyl, (un) substituted alkoxy, COR^{twenty one}Mosquito Ruboxy, carboalkoxy, OR^{twenty two}By lower chain alkyl or phenyl Carbamoyl, acyloxy, halo, cyano, and optionally substituted at nitrogen And azides.   R²⁰Represents the group: H, hydroxy, halo, lower chain alkyl, acyloxy, and Selected from siloxy;   Where R^{twenty one}Is a group: alkyl, alkenyl, alkynyl, aralkyl, (non ) Substituted phenyl, (un) substituted naphthyl, thienyl, furanyl and pyridyl Selected from;   And R^{twenty two}Is C_3-6Consists of a hydrocarbon moiety.   The compound of the general structure XIV is obtained by converting the ylide of the general structure XV to an acid chloride of the general structure XVI. Or reacting with an acid anhydride of general structure XVII:  Or an acid of general structure XVIII: With polyphosphoric acid, trifluoroacetic anhydride, or a similar reagent. What   Or chalcone of general structure XIX: Is treated with any base, or treated with a base, followed by 2,3- By treating with dichloro-5,6-dicyano-1,4-benzoquinone. Can be prepared,   Where the group R¹⁸, R¹⁹, R²⁰Is as described above, followed by withdrawal as needed Protection, coupling, addition, substitution, or elimination of any one or more groups R¹⁸, R¹⁹, R²⁰To a new group, R¹⁸, R¹⁹, R²⁰And, if desired, general The compound of formula XIV is converted to its salt or it is liberated from its salt.   Representative examples of compounds of general structure XIV are:     5,4'-dimethyl-7-acetylflavone     7-benzoyloxyflavanone     Apiin acetate.   Compounds of structure XX have the following formula:   Or a salt thereof,   here,   R^{twenty three}, R^{twenty four}, R^{twenty five}, R²⁶Is independently a group: H, hydroxy, (un) substituted alcohol Xy, siloxy, (un) substituted alkyl, (un) substituted alkenyl, halo, carboxy Selected from sil, and acyloxy, and wherein R^{twenty three}And R^{twenty four}And then And likewise R^{twenty five}And R²⁶Is a 5- to 7-membered (un) substituted carbocyclic ring or heterocyclic And a substituent on the optionally substituted group described above. Is lower chain alkyl, lower chain alkoxy, hydroxy, siloxy, acyloxy , Halo, benzoyl, carboxy, carboalkoxy, and lower chain alkyl, Optionally substituted at nitrogen by phenyl, thienyl, furyl, or pyridinyl It may include substituted carbamoyl.   Y¹Is a group: O, -OCH_TwoCH_TwoO-, -OCH_TwoCH_TwoS-, -OCH_TwoCH_Two CH_TwoO-, -SCH_TwoCH_TwoCH_TwoS- and -SCH_TwoCH_TwoSelected from S- ,   X^TwoIs a group: CH_Two, O, and S, with the following conditions: Do:   X and Y are O and R^{twenty four}Or R^{twenty five}Are both alkoxy Or alkoxy and alkyl (in no particular order)^{twenty three}And R²⁶Little At least one must be other than H.   Compounds of general structure XX can be prepared by converting an amide of general structure XXI to sec-butyl in THF. Reacting with lithium and tetramethylethylenediamine, followed by general Prepared by adding a benzaldehyde of structure XXII and adding an acid I can do it.   The resulting lactone of the general structure XXIII can be hydrogenated with a catalyst or activated in an acid. Can be reduced by treatment with zinc, followed by removal with trifluoroacetic anhydride. Hydrogenation or conversion of diaryls of the general structure XXIV to sulfuric acid, aluminum trichloride Treat with sodium, trifluoroacetic anhydride, or similar agents:   Where R^{twenty three}, R^{twenty four}, R^{twenty five}, R²⁶Is as defined above, followed by One if necessary, by deprotection, coupling, addition, substitution, or elimination The above group R^{twenty three}, R^{twenty four}, R^{twenty five}, R²⁶To a new group R^{twenty three}, R^{twenty four}, R^{twenty five}, R²⁶Convert to And, if desired, converting the compound of general structure XX to a salt thereof or converting it to a salt thereof. Release from its salt.   Particular representative examples of compounds of general structure XX include 3-isopropoxyanthrone No.   Another group useful in the invention is of the formula:  Or a salt thereof, wherein:   X^ThreeIs NR²⁷And X^FourIs CR³⁰And X^FiveIs O and X⁶Is CR³¹Is , X⁷Is O^-Is;   Or X^ThreeIs NR³⁰And X^FourIs CR²⁷Or N and X^FiveIs NR³¹In , X⁶Is CR²⁸And X⁷Is O^-Or S^-Is;   Or X^ThreeIs NR²⁷And X^FourIs CR³⁰And X^FiveIs NR²⁸And X⁶Is CR³¹And X⁷Is O^-Or S^-Is;   Or X^ThreeIs NR²⁷And X^FourIs CR²⁸Or N and X^FiveIs NR³⁰In , X⁶Is CR²⁹And X⁷Is NR³²Is;   Or X^ThreeIs NR³⁰And X^FourIs CR²⁷Or N and X^FiveIs NR²⁸In , X⁶Is CR²⁹And X⁷Is NR³²Is;   Or X^ThreeIs NR²⁷And X^FourIs CR³⁰And X^FiveIs S and X⁶Is CR³¹ And X⁷Is NR³²Is;   Or X^ThreeIs NR³⁰And X^FourIs CR²⁷And X^FiveIs S and X⁶Is CR²⁸ And X⁷Is NR³²Is;   Or X^ThreeIs S and X^FourIs CR³⁰And X^FiveIs NR²⁷And X⁶Is CR³¹ And X⁷Is O^-Or S^-Is;   Or X^ThreeIs S and X^FourIs CR³⁰And X^FiveIs NR²⁷And X⁶Is CR²⁸ And X⁷Is NR³²Is;   Or X^ThreeIs S and X^FourIs CR²⁷And X^FiveIs NR³⁰And X⁶Is CR²⁸ And X⁷Is NR³²Is;   Or X^ThreeIs S and X^FourIs CR³⁰And X^FiveIs S and X⁶Is CR²⁷so Yes, X⁷Is NR³²Is;   Or X^ThreeIs S and X^FourIs CR³⁰And X^FiveIs S and X⁶Is CR³¹so Yes, X⁷Is O^-Is;   Or X^ThreeIs NR³⁰And X^FourIs CR²⁷Or N and X^FiveIs NR³¹In , X⁶Is N and X⁷Is O^-Or S^-Is;   Or X^ThreeIs NR²⁷And X^FourIs CR³⁰And X^FiveIs NR²⁸And X^Four Is N and X⁷Is NR³²Or CZ^TwoZ^ThreeIs;   Or X^ThreeIs NR²⁷And X^FourIs CR²⁸Or N and X^FiveIs NR³⁰In , X^FourIs N and X⁷Is NR³²Or CZ^TwoZ^ThreeIs;   Or X^ThreeIs NR³⁰And X^FourIs N and X^FiveIs S and X⁶Is CR³¹so Yes, X⁷Is O^-Is;   Or X^ThreeIs S and X^FourIs CR²⁷And X^FiveIs NR³⁰And X⁶Is N Yes, X⁷Is NR³²Is;   Or X^ThreeIs S and X^FourIs CR³⁰And X^FiveIs NR²⁷And X⁶Is N Yes, X⁷Is NR³²Is;   Or X^ThreeIs O or S, X^FourIs N and X^FiveIs NR³⁰And X⁶Is N and X⁷Is NR³²Is;   here,   R²⁷, R²⁸, And R²⁹Is independently linear or branched lower alkyl ;   R³⁰And R³¹Is independently a group: hydrogen, straight or branched (un) substituted alkyl And (un) substituted aromatic systems, wherein the substituents may include: Halogen, linear or branched lower alkyl, alkenyl, 6-membered aromatic, cyclic alkyl Alkynyl, hydroxy optionally substituted by a killed or cyclic alkenyl ring , Linear or branched alkoxyl, benzoyl, carboalkoxy, lower Carbamoyl optionally substituted at the nitrogen by alkyl or phenyl, Or carboxy;   R³²Is a group: Ar, COAr, COR³³Where Ar is 6 members A (un) substituted aromatic ring wherein the substituents on this ring may include: Halogen, linear or branched lower alkyl, alkenyl, optionally 6-membered aromatic, Alkynyl, hydroxy substituted by cyclic alkyl or cyclic alkenyl ring Sil, straight or branched chain alkoxyl, benzoyl, carboalkoxy, lower chain Carbamoyl optionally substituted at the nitrogen by alkyl or phenyl, Or carboxy;   R³³Is selected from the group: hydrogen, and straight or branched chain alkyl;   Z^TwoAnd Z^ThreeIndependently represent the groups: CN and CO_TwoR³⁴Selected from;   R³⁴Is the group: hydrogen, linear or branched alkyl, and (un) substituted aromatic Selected from   The compound of general structure XXV is a compound of general structure XXVI:   (Where X⁸Is NR³⁰Or S and X⁹Is CR³⁰Or N and X^TenIs NR³⁰Or S and Z^FourIs CO_TwoH, CO_TwoR³⁰Or CN) is an acid salt By treating with a compound or anhydride,   Alternatively, a compound of general structure XXVII:   (Where X¹¹Is NR³⁰Or S and X¹²Is N or CR³⁰And X¹³Is Halogen, SMe, or OEt) with an amine, sulfur, or enolate Depending on the step of reacting,   Alternatively, a compound of general structure XXVIII:  (Where X¹⁵Is O or S) isocyanate, isothiocyanate, Or by reacting with carbon disulfide,   Or a compound of general structure XXIX: By reacting with sodium hydroxide,   Or a compound of general structure XXX: With alkyl tosylate, aryl tosylate or alkyl halide Depending on the process   Alternatively, a compound of general structure XXXI: With aryl diisocyanide, phosgene, thiophosgene, or 3,3-bis By reacting with (methylthio) acrylonitrile,   Or a compound of general structure XXXII:   (Where X¹⁶Is O, S, or NH) in the presence of acid chloride Reaction with ethoxide or HCl or with HCl in the presence of acid anhydride Depending on the process,   Or a compound of general structure XXXIII:   (Where X¹⁷Reacts NH or S) with acid chlorides, acid anhydrides or HONO Depending on the process Or a compound of general structure XXXIV: To Cu (acac)_TwoAnd reacting with   Where R^{twenty two}, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, And R³⁴Is determined above As defined, followed by deprotection, coupling, addition, substitution, Or one or more groups R by elimination^{twenty two}, R²⁸, R²⁹, R³⁰, R³¹, R³² , R³³, And R³⁴To a new group R^{twenty two}, R²⁸, R²⁹, R³⁰, R³¹, R³², R³³, And R³⁴And, if desired, converting the compound of general structure XXV to a salt thereof. Or liberate it from its salt.   Particular representative examples of compounds of general structure XXV include:   3- (4-chlorophenyl) -1,2,3,4-oxatriazolium-5- (4-chlorophenyl) aminide,   1,3-di (4-methylphenyl) -1,2,3,4-tetrazolium-5 Oxide. The following examples are intended to illustrate, but not limit, the invention.                                 Example 1   Compound 59-0008 was prepared according to McDonald, W.S. Chem Comm (1969) 392-393, Irving, H.N. Synthesized according to the procedure of N.H. et al., Anal Chim Acta (1970) 49: 261-266. Briefly Then, 10.0 g of dithizone was placed in 100 ml of Et0H and 50 ml of AcOH and heated under reflux for 18 hours. After cooling, it was first diluted with 100 ml of water, followed by addition of 50 ml of 1N NaOH. Followed It was further neutralized by adding 6N NaOH to a pH of 5.0. continue, The deep purple mixture was concentrated on a rotor vapor to remove organics. Once liquid When the purple color had completely disappeared, it was filtered to collect a dark precipitate. Hula Flash chromatography (4.5 × 25.7 cm, EtAc / Hep (1: 4), R_f0.22) And then recrystallized from EtOH to give 2.15 g of deep purple crystals (mp = 184-185 ° C.) Yield 25%). Example 2 High throughput screening   United States filed June 2, 1995 and incorporated herein by reference. Thousands of compounds were tested in the assay system described in 08 / 458.434. Standard positive The active control is a compound 59-0008 of the present invention of the following formula (also described as OS8): there were. :  More specifically, 2T3-BMP-2-LUC cells (stably transformed (see above) Ghosh-Choudhury et al., Endocrinology (1996) 137: 331-39)) was used. Cells were treated with α-MEM, 1% penicillin / streptomycin, and 1% glutamine ( Culture using 10% FCS with "plate medium") and split 1: 5 once a week Cracked. For the assay, cells were resuspended in plate medium containing 4% FCS and 5 × 10 on microtiter plate^ThreeConcentration of cells (in 50 μl) / well Plate at 5 ° C and 5% CO^TwoAnd incubated at 37 ° C for 24 hours. Up To begin the assay, add 50 μl of test compound or control in DMSO A 2 × concentration was added to the wells, resulting in a final volume of 100 μl. Final serum concentration Degree 2% FCS and final DMSO concentration was 1%. Compound 59-0008 (10 μM) Was used as a positive control.   Treated cells at 37 ° C for 24 hours, 5% CO^TwoIncubated in. Then the medium Removed and cells were rinsed three times with PBS. After removing excess PBS, 25 μl of 1 × fine Vesicle culture lysis reagent (Promega # E153A) is added to each well and at least 10 minutes Incubated. If necessary, plates / samples can be frozen at this point. You. Add 50 μl luciferase substrate (Promega # E152A; 10 ml Promega Lu Add 7mg Promega luciferase assay substrate) to the ciferase assay buffer. I got it. Fluorescence was measured in a 96-well automated fluorimeter and the fluorescence was Picogram of luciferase activity per microliter or microgram of protein It was shown as picogram of luciferase activity per mouse.   In this assay, compound 59-0008 (3-phenylazo-1H-4,1,2-benzo- Thiadiazine (3-phenylazo-1H-4,1,2-benzothiadiazine) is shown in FIG. Such a pattern of reactivity was shown. Activity against compound 59-0008 is approximately 3-10 μM At the concentration (more specifically at 3 μM), and therefore approximately 175 light emitting units (l ight emission unit). Therefore, other test compounds may be used at various concentrations. And these results were compared to the results obtained for 59-0008 at 10 μM (this Was normalized to 100). For example, in FIG. 2 and FIG. [0008] Any test compound that showed higher activity produced a value of 100 or more.   As shown in FIGS. 2 (39) and FIG. 3 (10), some compounds are particularly effective. It was found to be fruitful.                                 Example 3 In vivo calvarial bone growth data   Compound 59-0008 was prepared as described previously (“Compound on mouse calvarial bone growth” In vivo assay of the effect of, see above). Vehicle Compound 59-0008 quadruples the enhancement of new calvarial bone width compared to control Induced.Example 4 Chondrogenic activity   Compound 59-008, compound 59-0102, compound 50-0197, the effect of recombinant human BMP-2 In comparison, the effect on chondrocyte differentiation was assayed. In short, TMC-23, a mouse chondrogenic cell type, was converted to SV-40 large T-antigen. Mouse (Ghosh-Cho) containing the BMP-2 gene regulatory region udhury et al., Endocrinology 137: 331-39, 1996). It was isolated from bone cartilage and further cloned. These cells in DMEM / 10% FCS Cultured, and these cells show expression of the T antigen, and are also confluent Aggrecan (toluidine blue staining at pH 1.0) by 7 days after And the production of type II collagen (immunostaining).   For measurement of alkaline phosphatase (ALP) activity, LF Bonewald et al., J Biol.  Chem (1992) 267: 8943-49. Briefly, TMC-23 cells are 4 × 10 in a 96-well microtiter plate (in DMEM containing 10% FCS)^Threecell / Well. Two days after plating, the cells are confluent and Medium with fresh medium (10% FCS and different concentrations of compound or recombinant BMP-2 ). After an additional 2 or 5 days of incubation, plate Rinse twice with S, then add lysis solution (0.05% TrionX-100) (100 μl / well) Le). The cells are subjected to three freeze-thaw cycles (-70 ° C (30 minutes) followed by 37 ° C (with no shaking). 30 minutes)). 20 μl of cell lysate was added to 80 μl of 50 mM p-nitrophen 1.5 M 2-amino-2-methyl-propanol buffer with nil phosphate (pH 1 0.3) (Sigma ALP kit, Sigma Chemical Co., St. Louis, MO) . The reaction was stopped by adding 100 μl of 0.5 M NaOH. Spectral absorbance at 405nm Is compared with the standard absorbance of p-nitrophenol to evaluate the ALP activity in the sample did. Determine the protein content of the cell lysate using the Bio-Rad Protein Assay Kit ( Bio-Rad, Hercules, CA). Specific activity is determined by these two parameters. Calculated using data.   On the second day, compound 59-0008 (10^-9M), compound 59-0102 (10^-7M), and the compound 59-0197 (10^-9M) compared ALP levels with vehicle control about 3, 2, and 2.5 fold increase. At 100, 50, or 10 ng / ml, recombinant BMP2 Approximately 10, 4, or 1.5 times the ALP level, respectively, compared to vehicle control Was induced.   From the foregoing, it has been described that certain embodiments of the present invention have been described herein for purposes of illustration. Various modifications may be made without departing from the spirit and scope of the invention. Is recognized. Accordingly, the invention is not limited except as by the appended claims. Absent.

────────────────────────────────────────────────── ─── Continuation of front page    (81) Designated countries EP (AT, BE, CH, DE, DK, ES, FI, FR, GB, GR, IE, IT, L U, MC, NL, PT, SE), OA (BF, BJ, CF) , CG, CI, CM, GA, GN, ML, MR, NE, SN, TD, TG), AP (KE, LS, MW, SD, S Z, UG), EA (AM, AZ, BY, KG, KZ, MD , RU, TJ, TM), AL, AM, AU, BA, BB , BG, BR, CA, CN, CU, CZ, EE, FI, GE, HU, IL, IS, JP, KG, KP, KR, L C, LK, LR, LT, LV, MD, MG, MK, MN , MX, NO, NZ, PL, RO, SG, SI, SK, TR, TT, UA, UZ, VN (71) Applicant Board of Regents, The Univer             City of Texas System             United States Texas 78701, Oh             Stin, West 7 T             Treat 201 (72) Inventor Petri, Charles             United States Washington 98072, C             Woodinville, N. E. 196 tiers             Ichi Place 18459 (72) Inventor Olme, Mark W.             United States Washington 98117,             Attle, N. Wu. 98 Tea             Chi Street 636 (72) Inventor Baindur, Nando             United States Washington 98026, D             Domon's, 57 Tea H Place             Est 13919 (72) Inventors Robbins, Kirksey.             United States Washington 98055, Les             Ngong, Grant Avenue South             Number Y-304 1200 (72) Inventors Harris, Scott M.             United States Washington 98815,             Attle, 31 Estee Avenue             Nu. E. 6825 (72) Inventor Contoianni, Maria             United States Washington 98109,             Attle, Hays Street Number             504 769 (72) Inventor Huley, Lawrence H.             United States Texas 78731, Oh             Stin, Northwest Place             5915 (72) Inventor Curwin, Scene M.             United States Texas 78681, Lau             Ndlock, Ivy Court 703 (72) Inventor Mandy, Gregory Earl.             United States Texas 78230, Sun               Antonio, Morgan's Creek             3719

Claims (1)

[Claims] 1. A method of treating a condition in a vertebrate characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, said method requiring such treatment. Administering to the vertebrate subject to be treated an effective amount of a compound of the formula: Wherein R ^a is a non-interfering substituent; m is an integer from 0 to 4; each dashed line represents an arbitrary π-bond; Te N, NR, O, S, a CR or CR _2,, wherein each R is independently H or alkyl (1-6C); X is O, S, SO, or SO, be _2; L is an flexible linker; and Ar ² is a substituted or unsubstituted 6-membered aromatic ring, methods. 2. 2. The method of claim 1, wherein said L is a flexible conjugated linker. 3. Wherein L is a covalent bond, -N = N -, - RC = CR -, - RC = N -, - N = CR -, - NRCO -, - CONR -, - CR 2 O- and -CR ₂ NR- The method of claim 1, wherein each R is independently H or alkyl (1-6C). 4. Ar ² is The method of claim 1, wherein R ^b is a non-interfering substituent and n is an integer from 0-5. 5. Wherein Ar ² is an unsubstituted phenyl The method of claim 4. 6. Wherein said compound is 59-0008: The method of claim 1, wherein 7. A pharmaceutical composition for use in a method of treating a condition in a vertebrate, characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, wherein the composition comprises: Including a pharmaceutically acceptable excipient and an effective amount of a compound of the formula: Wherein R ^a is a non-interfering substituent; m is an integer from 0 to 4; each dotted line represents an arbitrary π-bond; each Z is independently N, NR, O, S, a CR or CR _2,, wherein each of R, are independently H or alkyl (l6C), X is, O, S, SO, or SO _2,, L Is a flexible linker; and Ar ² is a substituted or unsubstituted 6 membered aromatic ring. 8. The composition of claim 7, wherein L is a flexible conjugated linker. 9. Wherein L is a covalent bond, -N = N -, - RC = CR -, - RC = N -, - N = CR -, - NRCO -, - CONR -, - CR 2 O, and -CR ₂ NR- The composition of claim 7, wherein each R is independently H or alkyl (1-6C). 10. Ar ² is The composition of claim 7, wherein R ^b is a non-interfering substituent and n is an integer from 0 to 5. 11. The composition of claim 7, wherein Ar ² is unsubstituted phenyl. 12. Wherein said compound is 59-0008: The composition of claim 7, which is 13. A method of treating a condition in a vertebrate that is characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, the method comprising: Administering to the sample an effective amount of a compound of the formula: Here, R ^a is a located non-interfering substituents; n is an integer from 0 to 5; L is an flexible linker which does not contain nitrogen; and Ar ² is a substituted or unsubstituted phenyl or The method is a substituted or unsubstituted naphthyl. 14． Wherein R ^a is a -NR ₂ or -COOR, wherein, R is H or alkyl (l6C), The method of claim 13. 15. Wherein Ar ² is a substituted or unsubstituted phenyl The method of claim 13. 16. Ar ¹ and the Ar ² are different A method according to claim 13. 17． A pharmaceutical composition for use in a method of treating a condition in a vertebrate, characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, wherein the composition comprises: Including a pharmaceutically acceptable excipient and an effective amount of a compound of the formula: Here, R ^a is a located non-interfering substituents; n is an integer from 0 to 5; L is an flexible linker which does not contain nitrogen; and Ar ² is a substituted or unsubstituted phenyl or A composition which is substituted or unsubstituted naphthyl. 18. Wherein R ^a is a -NR ₂ or -COOR, wherein, R is H or alkyl (l6C), The composition of claim 17. 19. Wherein Ar ² is a substituted or unsubstituted phenyl, A composition according to claim 17. 20. Ar ¹ and the Ar ² are different, composition as claimed in claim 17. 21. A method of treating a condition in a vertebrate that is characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, the method comprising: Administering to the sample an effective amount of a compound of the formula: Where R ^a is a non-interfering substituent; n is an integer from 0 to 5; L is a constrained linker; and Ar ² is a substituted or unsubstituted phenyl or The method is a substituted or unsubstituted naphthyl. 22. Wherein R ¹ is -NR ^2, or -COOR, wherein, R is H or alkyl (l6C), The method of claim 21. 23. Wherein Ar ² is a substituted or unsubstituted phenyl The method of claim 21. 24. A pharmaceutical composition for use in a method of treating a condition in a vertebrate characterized by a defect in or a need for bone growth replacement and / or undesirable levels of bone resorption, wherein the composition comprises a pharmaceutical composition. Comprising a chemically acceptable excipient and an effective amount of a compound of the formula: Where R ^a is a non-interfering substituent; n is an integer from 0 to 5; L is a constrained linker; and Ar ² is a substituted or unsubstituted phenyl or a substituted or unsubstituted phenyl. A composition which is a substituted naphthyl. 25. Wherein R ¹ is -NR _2, or -COOR, wherein, R is H or alkyl (l6C), The composition of claim 24. 26. Wherein Ar ² is a substituted or unsubstituted phenyl, A composition according to claim 24. 27. The condition may be osteoporosis, fracture or bone loss, primary or secondary hyperparathyroidism, periodontal disease or defect, metastatic bone disease, osteolytic bone disease, after plastic surgery 22. The method according to any of claims 1, 13 or 21, wherein the method is after a prosthetic joint surgery or after a dental implantation. 28. 22. The method of any of claims 1, 13, or 21, further comprising administering to the subject one or more agents that promote bone growth or inhibit bone resorption. 29. 29. The method of claim 28, wherein the agent is selected from the group consisting of a bone morphogen, an anti-resorptive, an osteogenic factor, a cartilage-derived morphogenic protein, a growth hormone, and a differentiation factor.