Fluoro β-carboline compound

Abstract

In one aspect, the present invention provides compounds of Formula I, as well as salts, hydrates, and isomers thereof. In another aspect, the present invention provides methods for promoting bone formation in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound of Formula I. The present invention also provides orthopedic and periodontal devices, and methods for treating kidney disease, diabetes, bone loss, and cancer using compounds of Formula I.

Description

Technical Field

[0001] Cross - References to Related Applications This application claims the benefit of priority under 35 U.S.C.§119(e) of Title 119, United States Patent Law, to U.S. Provisional Application Serial No. 62 / 718,604, filed on August 14, 2018, the disclosure of which is hereby incorporated by reference in its entirety.

[0002] Statement Regarding Rights in Inventions Made Under Federally Sponsored Research and Development Not applicable

[0003] Reference to a "Sequence Listing", Table, or Computer Program Listings Appendix Submitted on a Compact Disk Not applicable

Background Art

[0004] Bone homeostasis involves a balanced process of bone formation and bone resorption. Increased bone resorption and loss of bone homeostasis are associated with many diseases and disorders such as osteoporosis and Paget's disease. All FDA - approved low - bone - density therapeutics except Teriparatide treat by stopping bone resorption and are thus bone resorption inhibitors. Bone resorption inhibitors act on osteoclasts by preventing them from resorbing bone.

[0005] It is known in the art that bone can be formed by two processes. One is mediated through a chondrocyte cartilage intermediate (endochondral), and the other is a direct process that stimulates osteoblasts (intramembranous). The endochondral process involves chondrocytes (chondrocyte / cartilage cell) dying and leaving voids that become occupied by mineralized osteoblasts on the surface of chondrocyte cartilage calcification. During the resorption process, osteoclasts resorb this cartilage calcification, leaving clean non-cartilaginous bone mineral. The endochondral process is present during the basic formation and growth period of long bones, and during the cartilage callus process period of fractures. The endochondral process begins when mesenchymal stem cells differentiate into chondrocytes to form cartilage. On the other hand, the intramembranous process occurs during the new bone growth stage of fractures and during the formation of the bones of the head. The intramembranous process occurs when mesenchymal stem cells differentiate into osteoblasts. Unlike cartilage, which is an elastic tissue, bone is hard and lacks flexibility. Two very different cell processes (osteoblasts vs chondrocytes) involving different molecules (WNT (Wingless and Int-1) vs BMP (bone morphogenetic protein)) and cell mechanisms (osteoblasts vs chondrocytes).

[0006] It is well understood that osteoblasts are involved in the secretion of bone mineral that causes an increase in bone density. To date, only teriparatide has been known to stimulate osteoblasts to increase mineral deposition, albeit indirectly via the Wnt pathway.

[0007] For the treatment of a variety of disorders such as simple aging, bone degeneration and osteoporosis, fracture healing, osteogenesis imperfecta, HPP, the healing of two bones or arthrodesis across a joint, low bone density disorders, etc. in mammals, and to successfully install various medical orthopedic implants and periodontal implants such as screws, rods, titanium cages for spinal fixation, hip joints, knee joints, ankle joints, shoulder joints, dental implants, bone grafts, plates and rods, etc., it is desirable to cause mineral deposition (bone formation) of osteoblasts.

[0008] For treating subjects in a low bone density state, but not limited to, the use of cathepsin K inhibitors, RankL inhibitors, denosumab, Prolia® (prolia), osteoprotegerin (OPG) inhibitors, alendronate, selective estrogen receptor modulators (SERMs), bisphosphonates, vitamin D, and other bone resorption inhibitors has resulted in at least a partially very small initial increase in bone mineral deposition of less than 6% in the first year, followed by a slight increase in subsequent years. The overall increase in bone density due to bone resorption inhibition therapy (cessation of bone loss) has been reported to be 9.4% over three years. Such treatable conditions may include osteopenia, osteoporosis, arthritis, tumor metastasis, osteogenesis imperfecta, Paget's disease, secondary low bone density disorders / diseases, and other metabolic bone disorders.

[0009] The use of parathyroid hormone and analogs, prostaglandin agonists, PDGE2, PDGE, Forteo® (forteo), osteoprotegerin (OPG) inhibitors, teriparatide, BMP2, BMP7, BMP4, EP4 agonists, etc. can be used to cause a desirable increase in bone mineral in subjects with the following low bone density states. Such states may include osteopenia, osteoporosis, arthritis, tumor metastasis, osteogenesis imperfecta, Paget's disease, secondary low bone density disorders, bone healing, spinal fusion, arthrodesis, and other metabolic bone disorders. However, the use of BMP agonists for treating systemic diseases has not progressed beyond Phase I clinical trials by the FDA.

[0010] Furthermore, the use of PTH, TGFβ binding proteins, etc. for increased bone mineralization to treat conditions that can be partially characterized by an increased risk of fractures such as osteopenia, degenerative disc disease, fractures, osteoporosis, arthritis, tumor metastasis, osteogenesis imperfecta, Paget's disease, and other metabolic bone disorders is known in the art. It is also known that demineralized bone matrix can partially promote a small amount of new bone growth by endogenous growth factors (TGFβ binding proteins (BMP)) that survive after the sterilization procedure of cadaver bone. However, demineralized bone matrix is usually supplied from a donor cadaver bank and is associated with certain risks such as disease transmission and bacterial contamination. Other versions of demineralized bone matrix are more severely processed and have fewer disease risks. The current unmet medical needs using currently approved treatments in the field of nonunion fractures (pseudarthrosis) require improvement of the nonunion observed in large defects of long bones consisting of large gaps between fracture ends. The use of demineralized bone or similar osteoconductive materials known in the art has not provided the desired effect of fusing long bones.

[0011] Accordingly, there is still a need in the art for new methods of treating bone disorders, fractures, and related problems. The present invention meets these and other needs. SUMMARY OF THE INVENTION

[0012] In one aspect, the present invention provides compounds and compositions, and methods of using such compounds and compositions. In a first aspect, the present invention provides a compound of formula I:

[0013]

Chemical formula

[0014] or a salt, hydrate, prodrug, or isomer thereof, wherein R 1a 、R 1b 、R 1c 、R 1d 、and R 2 are each independently hydrogen (H), halogen, C 1-6 alkyl, C1-6 Haloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, C 1-6 Alkyl-OH, -O-C 1-6 Alkyl-OH, C 3-6 Cycloalkyl-C 1-4 Independently selected from alkoxy, and hydroxyl group (-OH), provided that R 1a , R 1b , R 1c , R 1d , and R 2 Among them, no more than two are H, A is

[0015]

Chemical formula

[0016] and R N is selected from the group consisting of heterocyclyl and heteroaryl, The heterocyclyl moiety is selected from monocyclic, fused bicyclic, and bridged cyclic heterocyclyls. The monocyclic heterocyclyl contains 4 to 7 ring members. The fused bicyclic heterocyclyl and the bridged bicyclic heterocyclyl contain 7 to 10 ring members. Each heterocyclyl moiety has 1 to 3 heteroatoms as ring members selected from nitrogen (N), oxygen (O), and sulfur (S). Each heterocyclyl moiety contains at least one nitrogen atom as a ring member and is optionally substituted with 1 to 3 R 5 moieties, The heteroaryl moiety contains 5 to 10 ring members, at least one ring member is a nitrogen atom, and is optionally substituted with 1 to 3 R 5 moieties, and R 5 are each a hydroxyl group (-OH), C 1-3 alkyl, C 1-3 alkyl-OH, -O-C 1-3 alkyl, C 3-4 heteroalkyl, C 1-3Haloalkyl, -O-C 1-3 Provided is said compound selected from the group consisting of haloalkyl, halogen, and oxo.

[0017] In a second embodiment, the present invention provides a method for promoting bone formation and bone healing in a subject in need thereof. The method includes promoting bone formation in the subject by administering to the subject a therapeutically effective amount of a compound of Formula I described herein. The bone formation can be systemic or local. For local bone formation, in some embodiments, the compound can be administered with a bone-conductive agent, such as a bone-conductive matrix.

[0018] In a third embodiment, the present invention provides a method for treating kidney disorders. The method includes administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I.

[0019] In a fourth embodiment, the present invention provides a method for treating diabetes. The method includes administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I.

[0020] In a fifth embodiment, the present invention provides a method for treating cancer. The method includes administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I.

[0021] In a sixth embodiment, the present invention provides a medical device, such as an orthopedic or periodontal medical device. The medical device includes a structural support, and the implantable portion of the structural support is adapted to be permanently implanted within a subject. The implantable portion adheres to bone, and the structural support has at least a partial outer coating containing a compound of Formula I.

[0022] In a seventh embodiment, the present invention provides a compound or composition (e.g., a compound or composition of formula I) as described herein for use in the preparation of a medicament for the treatment of a disease or condition described herein. In some embodiments, the disease or condition is damaged bone, fracture, weakened bone, osteogenesis imperfecta, hypophosphatasia (HPP), osteopenia, osteoporosis, arthrodesis, or a condition characterized by low bone mass or low bone density. Also contemplated herein is the use of a compound or composition as described herein in a periodontal implant or a medical orthopedic implant. Orthopedic implants include screws, rods, and titanium cages used in spinal fixation, etc.

[0023] In an eighth embodiment, the present invention provides an orthobiologic, e.g., a bone formation inducer, for surgical implantation with or without a bone grafting device. The medical device includes a structural support, and the implantable portion of the structural support is adapted to be permanently implanted within a subject. The implantable portion adheres to bone, and the structural support has at least a partial outer coating containing a compound of formula I.

[0024] In a ninth embodiment, the present invention provides a method for treating osteopenia. The method includes administering to a subject in need thereof a therapeutically effective amount of a compound of formula I continuously or in combination with an osteoclast inhibitor. BRIEF DESCRIPTION OF THE DRAWINGS

[0025]

Figure 1

DETAILED DESCRIPTION OF THE INVENTION

[0026] I. Introduction Bone mass homeostasis and bone remodeling involve a balanced process of bone formation (osteoblast deposition of mineral, an anabolic process) and bone resorption (osteoclast resorption of mineral, bone loss, a catabolic process). These two processes are paired in healthy bone. See Figure 1. In bone formation, osteoblasts synthesize bone matrix and regulate mineralization, ultimately differentiating into osteocytes or bone lining cells. In bone resorption, another cell type, osteoclasts, remove mineralized bone matrix, degrade organic bone, and release calcium into the serum. See, for example, Kular et al., Clinical Biochemistry 45:863-873 (2012).

[0027] Osteoblasts (bone-forming cells) and osteoclasts (bone-resorbing cells) are controlled by various mechanisms. The differentiation of osteoclasts is controlled or regulated by osteoblasts (Glass et al., Dev Cell 8:751-764 (2005)) or other hormones such as PTH, calcitonin, and IL6. In contrast, the differentiation or activity of osteoblasts is not controlled or regulated by osteoclasts, but rather by various signals such as CPFA, hedgehog, WNT / LRP, and sclerostin. Bone formation can occur via endochondral ossification or intramembranous ossification (sclerostin). In intramembranous ossification, bone is formed directly by the stimulation of osteoblasts / osteocytes of bone cells. In endochondral ossification, bone formation occurs via a cartilage template, and the time until bone is formed becomes longer. BMP signaling is involved in endochondral ossification, whereas Wnt signaling has been shown to be involved in both endochondral ossification and intramembranous ossification.

[0028] In a normal healthy state, bone remodeling (or bone homeostasis) involves the breakdown of old bone (by osteoclasts) and the repair or replacement of old bone with new bone (by osteoblasts). When this homeostasis is disrupted and bone resorption exceeds bone formation, i.e., in a diseased bone state, as a result, bone resorption is detached from bone formation. An increase in bone resorption leads to bone loss (trabecular bone loss) and an increase in bone fragility (decrease in bone strength). Many diseases and conditions, including osteoporosis, osteogenesis imperfecta, Paget's disease of bone, metabolic bone diseases, bone changes secondary to cancer, and other diseases characterized by or associated with low bone density, are associated with an increase in bone resorption.

[0029] Diseases caused by increased bone resorption are associated with bone loss and increased bone fragility. To name just a few, they are often treated with bone resorption inhibitors such as bisphosphonates, denosumab, Prolia® (prolia), alendronate, cathepsin K modulators, RankL inhibitors, estrogen, cathepsin K inhibitors, and selective estrogen receptor modulators. These agents function by directly or indirectly preventing or inhibiting osteoclastic bone resorption. See Figure 1. However, these agents do not promote new bone formation (i.e., anabolic bone formation) by osteoblasts. In contrast, administration of a single dose of an anabolic agent usually results in a >8% annual cumulative increase from baseline in bone formation in the human lumbar spine. Administration of bone resorption inhibitors results in a slight increase in bone density of <7% in the first year, but subsequent increases in bone density are <3.5%, resulting in an annual cumulative increase of <10%. Therefore, bones with osteoporotic fragility treated with bone resorption inhibitors do not become more brittle, but since the bone resorption inhibitors increase bone density by depositing more bone mineral and do not promote new bone growth, the fragile bones do not become stronger or increase in strength. In contrast, for example, agents that promote anabolic bone growth by stimulating osteoblast activity either promote more bone matrix deposition or, when proliferation is stimulated, the agent results in more osteoblasts and thus more bone cells to bridge the gap and fuse two bones. Therefore, bones with osteoporotic fragility treated with anabolic bone formation agents prevent the bone from becoming more fragile and also enable the bone to have greater strength due to increased bone formation.

[0030] Referring to FIG. 1, without being bound by a particular theory, if the bone is considered as a bathtub, the drain would be associated with bone loss or resorption, and the faucet would be associated with added bone or bone formation. Due to aging or disease, both the faucet and the drain are added and removed at the same rate (in pairs) until the faucet bends or the size of the drain increases. Such perturbations cause an imbalance (dissociation) in formation / resorption, leading to a decrease in bone density. For example, imagine a sponge with an outer core and fibers extending from one end to the other on the inside. During bone resorption, these fibers are removed, and if bone resorption occurs at a faster rate than bone construction or formation, these fibers become fewer, and the bone becomes brittle. It doesn't take much force to break a sponge with fewer inner fibers compared to a sponge with more inner fibers. Since the process of bone resorption is well understood, many of the commercially available therapeutic agents act on osteoclasts to stop bone resorption. These include cathepsin K inhibitors, RankL inhibitors, denosumab, Prolia (registered trademark), Fosamax (registered trademark), EVISTA (registered trademark), Premarin (registered trademark), osteoprotegerin (OPG) inhibitors, alendronate, selective estrogen receptor modulators (SERMs), bisphosphonates, and other agents that act to stop the activity of osteoclasts.

[0031] Based on the example of the sponge, to increase the strength of the bone, the number of fibers inside the bone increases the strength of the bone. However, it is not possible to increase bone strength by acting on osteoclasts, which are bone resorbing cells. Therefore, it is necessary to focus on the bone that forms osteoblasts. Unlike bone resorption, bone formation is not well understood, and until recently, only one systemic therapeutic agent (teriparatide) and one surgical implant (injection of BMP protein) have been commercially available to promote bone formation. However, BMP products act to increase chondrocytes and promote cartilage production. In this process, chondrocytes may be replaced by osteoblasts.

[0032] Intermittent administration of teriparatide globally increases bone density by activating PKA, subsequently phosphorylating LRP and activating the WNT pathway (Wan et al., Genes Dev. 22(21): 2968-2979 (2008)). This increase in bone density occurs along the trabeculae already laid down within the bone matrix. Osteoblasts that reinforce the trabeculae secrete minerals into the existing trabecular bone, thus increasing the amount and density of minerals in the trabeculae.

[0033] When there are bone voids, whereby most of the bone is removed, causing non-union or a defect of significant size in the bone. The bone cannot heal itself across large gaps. Addition of BMP to this site causes pluripotent cells to differentiate into chondrocytes / cartilage, generating a cartilage callus. The ability of bone to fill the gap instead of cartilage is thought to require osteoblasts and osteocytes to undergo proliferation to fill the gap and then deposit minerals to fill the void.

[0034] Without being bound by a particular theory, the compounds of the present invention are believed to be SOST (sclerostin) antagonists and / or WISE antagonists that function by modulating the Wnt / LRP and / or BMP signaling pathways. SOST and WISE are proteins that are thought to regulate bone formation by binding to the Wnt receptor LRP to inhibit the Wnt signaling pathway or by binding to BMP and inhibiting BMP activity via different amino acid sequences or domains. By neutralizing the inhibitory effects of SOST protein and / or WISE protein on the Wnt pathway, the compounds and compositions of the present invention restore Wnt signaling and promote bone formation / growth. Thus, in one aspect, the present invention provides compounds, compositions, and methods for promoting bone formation in a subject. Bone formation can be systemic or local. The compounds and compositions of the present invention can be administered locally and / or systemically and, if desired, continuously and in combination with one or more other therapeutic agents. In another aspect, the present invention provides a device transplantable as a structural scaffold for promoting osteoblast / osteocyte migration to a scaffold and depositing bone mineral, and for delivering the compounds and compositions of the present invention, for example, for promoting bone formation at the transplant site. In another aspect, the compounds and compositions of the present invention can be used to treat kidney disorders, diabetes, osteopenia, and cancer.

[0035] II. Definitions As used herein, the term "pharmaceutically acceptable excipient" refers to a substance that aids in the administration of an active agent to a subject and its absorption by the subject. Pharmaceutically acceptable excipients useful in the present invention include, but are not limited to, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, and colorants. Those skilled in the art will recognize that other pharmaceutical excipients are useful in the present invention.

[0036] As used herein, the term "alkyl" refers to a straight-chain or branched saturated aliphatic radical having the indicated number of carbon atoms. For example, C1-C6 (or C 1-6) Alkyl includes, but is not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, iso - propyl, iso - butyl, sec - butyl, tert - butyl, etc.

[0037] Alkylene represents either a straight - chain or branched alkylene having 1 to 7 carbon atoms, that is, a divalent hydrocarbon radical having 1 to 7 carbon atoms. For example, straight - chain alkylene is a divalent radical of the formula -(CH2) n -, where n is 1, 2, 3, 4, 5, 6, or 7. Preferably, alkylene represents a straight - chain alkylene having 1 to 4 carbon atoms. For example, methylene, ethylene, propylene, or butylene chain, or methylene, ethylene, propylene, or butylene chain mono - substituted with C1 - C3 alkyl (preferably methyl) at the same or different carbon atoms, or disubstituted with C1 - C3 alkyl (preferably methyl) at the same or different carbon atoms, and the total number of carbon atoms is 7 or less. One of ordinary skill in the art will understand that a single carbon of alkylene can be divalent as in -CH((CH2) n CH3)-(n = 0 - 5).

[0038] As used herein, the terms "alkoxy" or "-O - alkyl" refer to an alkyl group containing an oxygen atom, for example, methoxy, ethoxy, etc. "Haloalkoxy" is as defined for alkoxy, where some or all of the hydrogen atoms are replaced by halogen atoms. For example, halo - substituted alkoxy includes trifluoromethoxy, etc.

[0039] The term "hydroxyalkyl" or "alkyl - OH (hydroxyl group)" refers to an alkyl group in which at least one of the hydrogen atoms is replaced by a hydroxy group as defined above. With respect to the alkyl group, the hydroxyalkyl group may have any suitable number of carbon atoms such as C 1-6 etc. Exemplary hydroxyalkyl groups include, but are not limited to, hydroxymethyl, hydroxyethyl (when hydroxy is in the 1 - or 2 - position), hydroxypropyl (when hydroxy is in the 1 -, 2 -, or 3 - position).

[0040] As used herein, the term "alkenyl" refers to any straight or branched hydrocarbon having from 2 to 6 carbon atoms and having at least one double bond. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, butenyl, isobutenyl, butadienyl, pentenyl, or hexadienyl.

[0041] As used herein, the term "alkynyl" refers to any straight or branched hydrocarbon having from 2 to 6 carbon atoms and having at least one triple bond. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, or butynyl.

[0042] As used herein, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0043] As used herein, the term "haloalkyl" refers to alkyl as defined above in which some or all of the hydrogen atoms are replaced by halogen atoms. Halogen (halo) preferably represents chloro or fluoro, but may also be bromo or iodo. For example, haloalkyl includes trifluoromethyl, fluoromethyl, etc. The term "perfluoro" defines a compound or radical having at least two available hydrogens replaced by fluorine. For example, perfluoromethane refers to 1,1,1-trifluoromethyl, and perfluoromethoxy refers to 1,1,1-trifluoromethoxy.

[0044] As used herein, the term "heteroalkyl" refers to an alkyl group having from 1 to 3 heteroatoms such as N, O, and S. A heteroalkyl group has the indicated number of carbon atoms, with at least one non-terminal carbon substituted with a heteroatom. Additional heteroatoms including, but not limited to, B, Al, Si, and P may also be useful. The heteroatoms may also be oxidized, such as, but not limited to, -S(O)- and -S(O)2-. For example, heteroalkyls include ethers, thioethers, and alkylamines. A heteroalkyl group does not include a peroxide (-O-O-) or other contiguous bonded heteroatoms.

[0045] As used herein, the term "oxo" refers to a double-bonded oxygen (=O).

[0046] As used herein, the term "cycloalkyl" refers to a saturated or partially unsaturated, monocyclic, fused bicyclic, or bridged polycyclic ring assembly containing from 3 to 12 ring atoms, from 3 to 8, from 3 to 6, or the indicated number of atoms. For example, C 3-8 Cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and up to cyclooctyl. The cycloalkyl groups of the present invention are optionally substituted as defined below.

[0047] As used herein, the terms "heterocyclic", "heterocycloalkyl", and "heterocyclyl" refer to ring systems having from 3 to about 20 ring members and from 1 to about 5 heteroatoms such as N, O, and S. Additional heteroatoms including, but not limited to, B, Al, Si, and P may also be useful. The heteroatoms may also be oxidized, including, but not limited to, -S(O)- and -S(O)2-. The term heterocyclic includes monocyclic, fused bicyclic, and bridged cyclic moieties. For example, heterocycles include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, morpholino, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperazinyl, piperidinyl, indolinyl, quinuclidinyl hexahydro-1H-furo[3,4-c]pyrrolyl, and 1,4-dioxa-8-aza-spiro[4.5]decan-8-yl. The heterocycloalkyl groups of the present invention are optionally substituted as defined below.

[0048] The substituents of the cycloalkyl groups and heterocyclyl groups are diverse and are independently selected from: the number of halogens within the range of 0 to the total number of open valences of the ring system, C 1-8 alkyl, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"C(O)2R', -NR'-C(O)NR"R"', -S(O)R', -S(O)2R', -S(O)2NR'R", phenyl, perfluoro(C1-C4)alkoxy, and perfluoro(C1-C4)alkyl (R', R", and R"' are independently selected from hydrogen, (C1-C8)alkyl, and C 3-8 heteroalkyl, and phenyl.

[0049] As used herein, a group "linked through a carbon atom" refers to a linkage between the carbon atom of the referenced group and the remainder of the molecule. A group "linked through a nitrogen atom" refers to a linkage between the nitrogen atom of the referenced group and the remainder of the molecule. By way of example only, a heterocyclyl group linked through a carbon atom may be

[0050]

Chem.

[0051] (the wavy line indicates the point of attachment to the remainder of the molecule). By way of example only, a heterocyclyl group linked through a nitrogen atom may be

[0052]

Chem.

[0053] (the wavy line indicates the point of attachment to the remainder of the molecule).

[0054] As used herein, when the referenced compound is an N-oxide, it contains an N-O bond having three additional bonds to nitrogen, i.e., an N-oxide is a group of R3N + -O - By way of example only, N-oxides may include

[0055]

Chem.

[0056] and the like.

[0057] As used herein, the term "aryl" refers to a monocyclic or fused bicyclic, tricyclic or higher aromatic ring assembly containing 6 to 16 ring carbon atoms. For example, aryl can be phenyl, benzyl, or naphthyl, preferably phenyl. "Arylene" means a divalent radical derived from an aryl group. An aryl group can be mono-substituted, di-substituted, or tri-substituted with one, two, or three radicals as described below.

[0058] The substituents of the aryl group are diverse and are selected from: - halogen, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', -CONR'R", -C(O)R', -OC(O)NR'R", -NR"C(O)R', -NR"C(O)2R', -NR'-C(O)NR"R"', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R", alkylenedioxy, heteroaryl, -C 1-2 alkylene-heteroaryl, heterocyclyl, C 1-2 alkylene-heterocyclyl, phenyl, perfluoro(C1-C4)alkoxy, and perfluoro(C1-C4)alkyl, (R', R", and R"' are independently selected from hydrogen, (C1-C8)alkyl, and C 3-8 heteroalkyl, and phenyl). Alkylenedioxy is a divalent substituent bonded to two adjacent carbon atoms of phenyl, such as methylenedioxy or ethylenedioxy. Oxy-C2-C3-alkylene is also a divalent substituent bonded to two adjacent carbon atoms of phenyl, such as oxyethylene or oxypropylene.

[0059] Examples of the substituted phenyl group include 4-chlorophen-1-yl, 3,4-dichlorophen-1-yl, 4-methoxyphen-1-yl, 4-methylphen-1-yl, 4-aminomethylphen-1-yl, 4-methoxyethylaminomethylphen-1-yl, 4-hydroxyethylaminomethylphen-1-yl, 4-hydroxyethyl-(methyl)-aminomethylphen-1-yl, 3-aminomethylphen-1-yl, 4-N-acetylaminomethylphen-1-yl, 4-aminophen-1-yl, 3-aminophen-1-yl, 2-aminophen-1-yl, 4-phenyl-phen-1-yl, 4-(imidazol-1-yl)-phenyl, 4-(imidazol-1-ylmethyl)-phen-1-yl, 4-(morpholin-1-yl)-phen-1-yl, 4-(morpholin-1-ylmethyl)-phen-1-yl, 4-(2-methoxyethylaminomethyl)-phen-1-yl, and 4-(pyrrolidin-1-ylmethyl)-phen-1-yl, 4-(thiophenyl)-phen-1-yl, 4-(3-thiophenyl)-phen-1-yl, 4-(4-methylpiperazin-1-yl)-phen-1-yl, and 4-(piperidinyl)-phenyl and 4-(pyridinyl)-phenyl, but are not limited thereto.

[0060] As used herein, the term "heteroaryl" refers to a monocyclic, fused bicyclic, or tricyclic aromatic ring assembly containing 5 to 16 ring atoms, wherein 1 to 4 of the ring atoms are each a heteroatom of N, O, or S. For example, heteroaryl includes pyridyl, indolyl, indazolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothienyl, benzofuranyl, furanyl, pyrrolyl, thiazolyl, benzothiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, or other radicals substituted, particularly mono- or disubstituted, for example by alkyl, nitro, or halogen. Pyridyl represents 2-, 3-, or 4-pyridyl. Thienyl represents 2- or 3-thienyl. Quinolinyl preferably represents 2-, 3-, or 4-quinolinyl. Isoquinolinyl preferably represents 1-, 3-, or 4-isoquinolinyl. Benzopyranyl and benzothiopyranyl each preferably represent 3-benzopyranyl or 3-benzothiopyranyl, respectively. Thiazolyl preferably represents 2- or 4-thiazolyl, most preferably 4-thiazolyl. Triazolyl is preferably 1-, 2-, or 5-(1,2,4-triazolyl). Tetrazolyl is preferably 5-tetrazolyl. As defined below, the heteroaryl moiety may optionally be substituted.

[0061] Preferably, heteroaryl is pyridyl, indolyl, quinolinyl, pyrrolyl, thiazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, thienyl, furanyl, benzothiazolyl, benzofuranyl, isoquinolinyl, benzothienyl, oxazolyl, indazolyl, or any other radical that is substituted, particularly mono- or disubstituted.

[0062] The substituents of the heteroaryl group are diverse and are selected from the following: halogen in an amount within the range of 0 to the total number of open valences of the heteroaromatic ring system, -OR’, -OC(O)R’, -NR’R”, -SR’, -R’, -CN, -NO2, -CO2R’, -CONR’R”, -C(O)R’, -OC(O)NR’R”, -NR”C(O)R’, -NR”C(O)2R’, -NR’-C(O)NR”R”’, -S(O)R’, -S(O)2R’, -S(O)2NR’R”, perfluoro(C1-C4)alkoxy, and perfluoro(C1-C4)alkyl (R’, R”, and R”’ are independently selected from hydrogen, (C1-C8)alkyl, C 3-8 heteroalkyl, and phenyl).

[0063] As used herein, the terms “ring member” and “ring vertex” are intended to have the same meaning. For example, a 6-membered ring has 6 ring vertices.

[0064] As used herein, “C 3-6 cycloalkyl-C 1-4 alkoxy” refers to an alkoxy group in which one hydrogen atom is substituted with a C 3-6 cycloalkyl group.

[0065] As used herein, the term “salt” refers to an acidic or basic salt of a compound used in the methods of the present invention. Exemplary examples of pharmaceutically acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid, etc.) salts, quaternary ammonium (methyl iodide, ethyl iodide, etc.) salts. Pharmaceutically acceptable salts are understood to be non-toxic. Additional information regarding suitable pharmaceutically acceptable salts is described in Remington’s Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, which is incorporated herein by reference.

[0066] The pharmaceutically acceptable salts of the acidic compounds of the present invention are salts formed with bases, i.e., cationic salts such as alkali and alkaline earth metal salts of sodium, lithium, potassium, calcium, magnesium, etc., and ammonium salts such as ammonium, trimethylammonium, diethylammonium, tris(hydroxymethyl)-methyl-ammonium salts.

[0067] Similarly, acid addition salts of mineral acids, organic carboxylic acids, and organic sulfonic acids, such as hydrochloric acid, methanesulfonic acid, maleic acid, etc., can also be provided, and basic groups such as pyridyl form part of the structure.

[0068] The neutral form of the compound can be regenerated by contacting the salt with a base or an acid and isolating the parent compound by conventional methods. The parent form of the compound differs from the various salt forms in certain physical properties such as solubility in polar solvents, but otherwise, the salts are equivalent to the parent form of the compound for the purposes of the present invention.

[0069] As used herein, the term "calcium salt" refers to a salt containing calcium. Examples of calcium salts include, but are not limited to, calcium acetate, calcium aluminate, calcium aluminate, calcium arsenide, calcium borate, calcium bromide, calcium carbide, calcium carbonate, calcium chloride, calcium chloride, calcium citrate, calcium citrate malate, calcium cyanamide, calcium dihydrogen phosphate, calcium fluoride, calcium formate, calcium gluceptate, calcium glucoheptonate, calcium gluconate, calcium glycerophosphate, calcium hexaborate, calcium hydride, calcium hydroxide, calcium hypochlorite, calcium inosinate, calcium iodate, calcium iodide, calcium lactate, calcium lactate gluconate, calcium magnesium acetate, calcium malate, calcium nitrate, calcium nitride, calcium oxalate, calcium oxide, calcium pangamate, calcium peroxide, calcium phosphate, calcium phosphide, calcium propionate, calcium pyrophosphate, calcium silicate, calcium silicide, calcium sorbate, calcium stearate, calcium sulfate, calcium sulfide, calcium tartrate, calcium chloride (I), dicalcium citrate, dicalcium phosphate, dodecacalcium hepta-aluminate, tricalcium aluminate, tricalcium phosphate, and triple superphosphate. Those skilled in the art will understand that other calcium salts may be useful in the present invention.

[0070] As used herein, the term "hydrate" refers to a compound that forms a complex with at least one water molecule. The compounds of the present invention can form complexes with 1 to 10 water molecules. The term "hydrate" also includes hemihydrates in which there are two compounds for each water molecule of the complex.

[0071] Certain compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds, and it is intended that all racemates, diastereomers, geometric isomers, and individual isomers are included within the scope of the present invention.

[0072] As used herein, the term "subject" refers to an animal such as a mammal including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice, etc. In certain embodiments, the subject is a human.

[0073] As used herein, the terms "therapeutically effective amount or dose", "therapeutically sufficient amount or dose", or "effective or sufficient amount or dose" refer to an amount or dose that produces a therapeutic effect when administered. The exact amount will be determined by the purpose of the treatment and can be ascertained by one of ordinary skill in the art using known methods (see, e.g., Lieberman, Pharmaceutical Dosage Forms (vols. 1 - 3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999), Pickar, Dosage Calculations (1999), and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins). In sensitized cells, the therapeutically effective amount is often lower than the conventional therapeutically effective amount for non - sensitized cells.

[0074] As used herein, the term "site of injury or local condition" refers to a specific location within the body of a subject in need of treatment by the methods of the present invention. For example, the injury may be a fracture, and the local condition may be a medical condition (such as osteoporosis) limited to a specific location of the subject's body, such as a particular bone, joint, finger, hand, foot, limb, spine, head, torso, etc. In some embodiments, the site of injury or local condition is a surgical implantation site.

[0075] As used herein, the term "promotion of bone formation" refers to the stimulation of new bone formation, the growth of bone across a joint or gap, the enhancement or promotion of bone formation, and / or an increase in bone density or bone mineral content. In some embodiments, a compound promotes bone formation if it increases the amount of bone in a sample by at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20% or more compared to a control sample (e.g., a sample not in contact with the compound).

[0076] As used herein, the term "arthrodesis" often refers to the artificial induction of joint ossification across two bones and / or a joint, often via surgery. Arthrodesis can be achieved, inter alia, via the use of bone grafts, metal implants, or artificial bone substitutes.

[0077] As used herein, the term "bone autograft" refers to the transplantation of a subject's own bone.

[0078] As used herein, the term "bone allograft" refers to the transplantation of bone from one person to another.

[0079] As used herein, the term "bone resorption inhibitor" refers to a drug that delays or blocks bone resorption and / or acts on osteoclasts.

[0080] As used herein, the term "bone-related disease characterized by low bone mass" refers to bone with a T-score of less than -0.5. Other methods of determining low bone mass are known to those of skill in the art.

[0081] As used herein, the term "fracture" refers to a crack, break, or damaged bone at one or more locations along the bone. In some embodiments, the term "fracture" also includes segments of bone that are missing.

[0082] As used herein, the term "spinal arthrodesis" refers to a surgical procedure for joining or fusing two or more vertebrae.

[0083] As used herein, the term "structural support" refers to a segment (transplantable portion) of a device that can be implanted into a subject. The structural support can be prepared from a variety of different materials, including metals, ceramics, polymers, and inorganic materials as described below. The structural support may be coated with various materials that promote bone growth. In some embodiments, the entire device comprises a structural support that is implantable. For example, in some embodiments, the entire device described herein can be implanted into a surgical site and the surgical site can be closed over the device.

[0084] As used herein, the term "outer coating" refers to a coating of the structural support that can cover only a portion of the structural support (partial outer coating) or can cover the entire structural support. For example, a partial outer coating can completely cover only the implantable portion of the structural support.

[0085] As used herein, the terms "weakened bone", "low bone density", or "low bone mass" refer to bone having a T-score of less than -0.5 (less than 0.9 g / cm 2 2).

[0086] As used herein, the term "demineralized bone" refers to bone from which inorganic minerals have been removed. The remaining organic collagen material may contain bone-inductive growth factors. These growth factors include bone morphogenetic proteins that induce cartilage, which ossifies via endochondral ossification to produce new bone formation. Demineralized bone is often provided in the form of "demineralized bone matrix (DBM)". DBM can be made from fresh frozen or lyophilized bulk bone allografts or from gentle acid extraction of cadaver bone that removes the mineral phase leaving collagen, growth factors, and non-collagenous proteins that provide the inherent properties of osteoconduction. DBM may be processed in various ways and ultimately the powder is mixed with a carrier to provide optimal handling properties desirable for the surgeon. DBM is clinically available in gels, pastes, putties, and fabrics tailored to meet the needs of surgical procedures. Some DBMs are mixed with antibiotics prior to surgical procedures.

[0087] As used herein, the term "kidney disorder" refers to the inability of the kidneys to excrete waste products and to help maintain the body's electrolyte balance. Kidney disorders are characterized by several of the following: high blood pressure, accumulation of urea and formation of urea frost, accumulation of potassium in the blood, decreased erythropoietin synthesis, increased fluid volume, hyperphosphatemia, and metabolic acidosis, among others.

[0088] As used herein, the term "diabetes" refers primarily to a condition characterized by the inability to produce a sufficient amount of the hormone insulin, which is produced by the pancreas. When insulin is released into the bloodstream, it induces glucose uptake by cells. Therefore, when the amount of insulin is insufficient, the blood glucose level of the affected individual rises. Those skilled in the art will recognize that the inability of the body to produce a sufficient amount of insulin can be a feature of both type 1 and type 2 diabetes.

[0089] As used herein, the term "osteoconductive matrix" refers to a material having a scaffold structure that can act as an osteoconductive substrate (i.e., enable bone growth), to which infiltrating cells can attach, proliferate, and participate in the process of generating osteoid (the organic phase of bone) leading to bone neoformation or new bone formation. The terms "matrix" and "scaffold" are used interchangeably to refer to a structural component or substrate that essentially has a three-dimensional form in which specific cellular events involved in bone formation occur. The osteoconductive matrix allows for the ingrowth of host capillaries, perivascular tissue, and bone progenitor cells. In some embodiments, the osteoconductive matrix contains an "osteogenic agent" to provide osteogenic ability. As used herein, an osteogenic agent is an agent that stimulates the host to proliferate bone cells, thereby producing more bone osteoid.

[0090] As used herein, the terms "treat", "treating", and "treatment" refer to signs of success in the treatment or amelioration of an injury, condition, state, or symptom (e.g., pain) that include the following objective or subjective parameters: reduction of symptoms, remission, alleviation, or making the symptom, injury, condition, or state more tolerable to the patient, decreasing the frequency or duration of the symptom or condition, or, depending on the situation, preventing the onset of the symptom or condition. Treatment or amelioration of a symptom can be based on objective or subjective parameters including, for example, the results of a physical examination.

[0091] As used herein, the term "RankL inhibitor" refers to a compound or agent that inhibits the activity of RankL. RankL (receptor activator of nuclear factor κB ligand) is important for bone metabolism by activating osteoclasts. RankL inhibitors include, but are not limited to, the human monoclonal antibody denosumab. Those skilled in the art will understand that other RankL inhibitors are useful in the present invention.

[0092] As used herein, the terms "parathyroid hormone" or "PTH" refer to a compound or agent that acts on the PTH receptor to activate a pathway. PTH is important for bone metabolism by activating osteoblasts. PTH includes, but is not limited to, teriparatide, Forteo® (forteo), and abaloparatide-SC. Those skilled in the art will understand that other PTHs may be useful in the present invention.

[0093] As used herein, the term "combination therapy" means using the present invention in combination, together, or sequentially, before or after administration of a compound of the present invention.

[0094] Certain compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds, and racemates, diastereomers, geometric isomers, positional isomers, and individual isomers (e.g., separate enantiomers) are all intended to be included within the scope of the present invention. In some embodiments, the compounds of the present invention are specific enantiomers or diastereomers that are substantially free of other forms. The compounds of the present invention may also contain unnatural proportions of atomic isotopes in one or more of the atoms that make up such compounds. For example, the compound may be radiolabeled with radioactive or non-radioactive isotopes such as, for example, deuterium ( 2 H), tritium ( 3 H), iodine-125 ( 125 I), or carbon-14 ( 14 C). All isotopic variants of the compounds of the present invention are intended to be included within the scope of the present invention, whether radioactive or not.

[0095] III. Compounds and Compositions In some embodiments, the present invention provides a compound according to the following formula I:

[0096]

Chemical formula

[0097] or a salt, hydrate, prodrug, or isomer thereof, provided that R 1a 、R 1b 、R 1c 、R 1d 、and R 2 are each independently selected from hydrogen (H), halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkyl-OH, -O-C 1-6 alkyl-OH, C 3-6 cycloalkyl-C 1-4 alkoxy, and hydroxyl (-OH), provided that no more than two of R 1a 、R 1b 、R 1c 、R 1d 、and R 2 are H, A is

[0098]

Chemical formula

[0099] and R N is selected from the group consisting of heterocyclyl and heteroaryl, the heterocyclyl moiety is selected from monocyclic, fused bicyclic, and bridged cyclic heterocyclyls, the monocyclic heterocyclyl contains 4 to 7 ring members, the fused bicyclic heterocyclyl and the bridged bicyclic heterocyclyl contain 7 to 10 ring members, each heterocyclyl moiety has 1 to 3 heteroatoms as ring members selected from nitrogen (N), oxygen (O), and sulfur (S), each heterocyclyl moiety contains at least one nitrogen atom as a ring member, and optionally is substituted with 1 to 3 R 5 moieties, the heteroaryl moiety contains 5 to 10 ring members, at least one ring member is a nitrogen atom, and optionally is substituted with 1 to 3 R5 is partially replaced, and R 5 are each selected from the group consisting of a hydroxyl group (-OH), C 1-3 alkyl, C 1-3 alkyl-OH, -O-C 1-3 alkyl, C 3-4 heteroalkyl, C 1-3 haloalkyl, -O-C 1-3 haloalkyl, halogen, and oxo, said compound.

[0100] In some embodiments, R 1a , R 1b , R 1c , and R 1d are each independently selected from the group consisting of hydrogen (H), halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy, provided that no more than two of R 1a , R 1b , R 1c , and R 1d are H.

[0101] In some embodiments, R 1a , R 1b , R 1c , and R 1d are each independently selected from the group consisting of hydrogen (H), halogen, and C 1-6 alkoxy, provided that no more than two of R 1a , R 1b , R 1c , and R 1d are H.

[0102] In some embodiments, R 1a , R 1b , R 1c , and R 1d are each independently selected from the group consisting of hydrogen (H), fluorine (F), and methoxy, provided that no more than two of R 1a , R 1b , R 1c , and R1d Two or fewer of them are H.

[0103] In some embodiments, R of formula I 1c is H or F. In some embodiments, R of formula I 1d is H. In some embodiments, R of formula I 1b is C 1-6 alkoxy. In some embodiments, R of formula I 1b is methoxy. In some embodiments, R of formula I 1a is halogen. In some embodiments, R of formula I 1a is F.

[0104] In some embodiments, R of formula I 2 is H, C 1-6 alkyl, or C 1-6 haloalkyl. In some embodiments, R of formula I 2 is H or C 1-6 alkyl. In some embodiments, R of formula I 2 is C 1-6 haloalkyl. In some embodiments, R of formula I 2 is CF3. In some embodiments, R of formula I 2 is CH3.

[0105] In some embodiments, R of formula I N is monocyclic, fused bicyclic, or bridged heterocyclyl. In some embodiments, R of formula I N is monocyclic heterocyclyl. In some embodiments, said monocyclic heterocyclyl is a piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, or azetidinyl group optionally substituted with one or two R 5 moieties. In some embodiments, R of formula I N is fused bicyclic or bridged heterocyclyl. In some embodiments, said fused bicyclic or bridged heterocyclyl is

[0106]

Chemical formula

[0107] selected from each having 1 to 2 Rs 5 optionally replaced at the desired position.

[0108] In some embodiments, when R N is a condensed bicyclic or bridged heterocyclyl, R 5 is -OH, C 1-3 alkyl, or -O-C 1-3 alkyl.

[0109] In some embodiments, R N of formula I is heteroaryl. In some embodiments, the heteroaryl moiety contains 5 to 8 ring members, at least one ring member is a nitrogen atom, and is optionally substituted with 1 to 3 R 5 moieties. In some embodiments, R N of formula I is a tetrazolyl, imdidazolyl, or pyridinyl group optionally substituted with 1 to 3 R 5 moieties.

[0110] In some embodiments, R N of formula I is

[0111]

Chemical formula

[0112] selected from the group consisting of.

[0113] In some embodiments, R N of formula I is

[0114]

Chemical formula

[0115] selected from the group consisting of.

[0116] In some embodiments, R of formula I N is

[0117]

Chem.

[0118] selected from the group consisting of.

[0119] In some embodiments, R of formula I N is

[0120]

Chem.

[0121] is.

[0122] In some embodiments, R 5 is -OH, C 1-3 alkyl, C 1-3 alkyl-OH, -O-C 1-3 alkyl, C 1-3 haloalkyl, -O-C 1-3 haloalkyl, halogen, and oxo. In some embodiments, R 5 is -OH, C 1-3 alkyl, or -O-C 1-3 alkyl.

[0123] In some embodiments, the present invention provides a compound according to the following formula I:

[0124]

Chem.

[0125] or a salt, hydrate, prodrug, or isomer thereof, wherein R 1a and R 1b are each independently hydrogen (H), halogen, C1-6 Alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, and C 1-6 independently selected from the group consisting of haloalkoxy, R 1c and R 1d are independently H or halogen, provided that R 1a , R 1b , R 1c , and R 1d among which no more than two are H, R 2 is C 1-6 alkyl or C 1-6 haloalkyl, A is

[0126]

Chemical formula

[0127] and R N is a heterocyclyl optionally substituted with 1 to 3 R 5 substituents, and the heterocyclyl moiety is a monocyclic, bridged bicyclic, or fused bicyclic heterocycle, and R 5 is as defined by formula I.

[0128] In some embodiments, the present invention provides a compound according to the following formula I:

[0129]

Chemical formula

[0130] or a salt, hydrate, prodrug, or isomer thereof, wherein R 1a and R 1b are each independently hydrogen (H), halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6Independently selected from the group consisting of haloalkoxy, R 1c and R 1d are independently H or halogen, provided that R 1a R 1b R 1c and R 1d two or less of are H, R 2 is C 1-6 alkyl or C 1-6 haloalkyl, A is

[0131]

Chemical formula

[0132] and R N is a heterocyclyl optionally substituted with 1 to 3 R 5 substituents, and the heterocyclyl moiety is a monocyclic, bridged bicyclic, or fused bicyclic heterocycle, and R 5 is as defined in Formula I and the secondary embodiments described herein.

[0133] In some embodiments, the present invention provides a compound of Formula I having the following structure:

[0134]

Chemical formula

[0135] or a salt, hydrate, prodrug, or isomer thereof, wherein R 1a is halogen, R 1b is C 1-6 alkoxy, R 1c and R 1d are independently hydrogen (H) or fluorine (F), R 2 is C1-6 Haloalkyl or C 1-6 is alkyl, and A is

[0136]

Chemical formula

[0137] and R N is

[0138]

Chemical formula

[0139] selected from the group consisting of.

[0140] In some embodiments of the compound of formula I, each variable region is as defined in the preceding paragraph, except for R N and is as follows:

[0141]

Chemical formula

[0142] In some embodiments of the compound of formula I, each variable region is as defined in the preceding paragraph, except for R N and is as follows:

[0143]

Chemical formula

[0144] In a group of some embodiments, a compound of formula I having a structure selected from the following:

[0145]

Chemical formula

[0146] or a salt, hydrate, or prodrug thereof.

[0147] In some embodiments, the present invention provides formates of the compounds by any of the above compounds. In some embodiments, the present invention provides citrates of the compounds by any of the above compounds. In some embodiments, the present invention provides hydrochlorides of the compounds by any of the above compounds.

[0148] The compounds and compositions of the present invention may also include hydrates, solvates, and prodrug forms. The compounds and compositions of the present invention may also optionally include isomers and metabolites of the compounds of Formula I.

[0149] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound according to Formula I and a pharmaceutically acceptable excipient.

[0150] The compounds of the present invention can be in the form of salts. Salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, phosphonate, isonicotinate, lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Other salts include, but are not limited to, salts with inorganic bases including alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, aluminum salts, and ammonium salts. Other salts containing organic bases include salts containing diethylamine, diethanolamine, meglumine, and N,N'-dibenzylethylenediamine. In some embodiments, the present invention provides the aforementioned hydrochloride.

[0151] In some embodiments, the compounds of the invention contain nitrogen atoms that are optionally further oxidized, i.e., the compounds are N-oxides. By way of mere example, as an example, the nitrogen atom of the pyrido-indolyl ring system in the compounds of formula (I) is oxidized to the corresponding N-oxide.

[0152] In some embodiments, the compounds described herein are delivered and / or formulated as prodrugs. In one embodiment, any of the compounds described herein is an ester prodrug. In another embodiment, any of the compounds described herein is an amide prodrug. In a further embodiment, the prodrug moiety contains a conjugate group that allows for selective targeting at the bone structure. Examples of such motifs are described in Erez et al., Bioorg. Med. Chem. Lett. 2008, 18, 816 - 820 and Neale et al., Bioorg. Med. Chem. Lett. 2009, 19, 680 - 683, which are incorporated herein by reference. Accordingly, within the scope of the embodiments presented herein are estradiol conjugates and / or bisphosphonate conjugates of the compounds of formula I.

[0153] The compounds of the invention can be made using various synthetic methods known to those skilled in the art (see Comprehensive Organic Transformations Richard C. Larock, 1989). Those skilled in the art will understand that other methods of making the compounds are useful in the present invention. Exemplary methods for synthesizing the compounds of formula I are described in the Examples section and in Scheme 1 below. Scheme 1

[0154]

Chemical formula

[0155] Starting from Compound 1, the reaction with Compound 2 containing a leaving group (LG) provides a compound of Formula I. Various leaving groups are suitable, including but not limited to halo, activated ester, mesylate, triflate, or any other suitable leaving group that enables the attachment of a group at the 9-position of the core ring system. Optionally, when R

[0156]

Chemical formula

[0157] is methoxy, it may be converted to a hydroxy group by demethylation using the procedures described, for example, HBr in acetic acid, boron tribromide, or any other suitable procedure. Optionally, the compound of Formula I includes an N-oxide prepared, for example, by oxidation using chloro perbenzoic acid. 1b If R

[0158] IV. Method for promoting bone formation In another aspect, the present invention provides a method for promoting bone formation and bone healing in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound of the present invention (e.g., a compound or composition of Formula I as described in Section III above).

[0159] In some aspects, the present invention is a method for promoting in a subject in need of bone formation, comprising administering to the subject a therapeutically effective amount of a compound of the following Formula I:

[0160]

Chemical formula

[0161] or a salt, hydrate, prodrug, or isomer thereof, the method comprising R 1a 、R 1b 、R 1c 、R 1d 、and R 2 are each independently hydrogen (H), halogen, C 1-6 alkyl, C 1-6Haloalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy, C 1-6 Alkyl-OH, -O-C 1-6 Alkyl-OH, C 3-6 Cycloalkyl-C 1-4 Independently selected from alkoxy and hydroxyl group (-OH), provided that R 1a , R 1b , R 1c , R 1d , and R 2 Among them, no more than two are H, A is

[0162]

Chemical formula

[0163] and R N is selected from the group consisting of heterocyclyl and heteroaryl, The heterocyclyl moiety is selected from monocyclic, fused bicyclic, and bridged cyclic heterocyclyls. The monocyclic heterocyclyl contains 4 to 7 ring members. The fused bicyclic heterocyclyl and the bridged bicyclic heterocyclyl contain 7 to 10 ring members. Each heterocyclyl moiety has 1 to 3 heteroatoms as ring members selected from nitrogen (N), oxygen (O), and sulfur (S). Each heterocyclyl moiety contains at least one nitrogen atom as a ring member and is optionally substituted with 1 to 3 R 5 moieties, The heteroaryl moiety contains 5 to 10 ring members, at least one ring member is a nitrogen atom, and is optionally substituted with 1 to 3 R 5 moieties, and each R 5 is -OH, C 1-3 alkyl, C 1-3 alkyl-OH, -O-C 1-3 alkyl, C 3-4 heteroalkyl, C 1-3 haloalkyl, -O-C1-3 Providing the method selected from the group consisting of haloalkyl, halogen, and oxo.

[0164] In some embodiments, the method comprises administering to a subject a therapeutically effective amount of a hydrochloride, sulfate, formate, or citrate salt of a compound of Formula I above.

[0165] One of ordinary skill in the art will understand that mineral deposition (bone formation) in osteoblasts can be achieved by local administration, systemic administration, or both local and systemic administration. In some embodiments, bone formation is local. Subjects in need of local bone formation can have any of a variety of diseases or conditions (including, but not limited to, diseases or conditions characterized by weakened bone, fractures, or low bone mass or insufficient mineralization as described herein). In some embodiments, the subject is in need of spinal fusion, bone healing, arthrodesis, or orthopedic, dental, or periodontal synthetic bone grafts or implants. In some embodiments, the present invention provides a method of promoting bone formation at the site of an injury or a local condition. In some embodiments, the present invention includes a method of fusing bone (e.g., at the site of an injury). In some embodiments, the injury site is a surgical site. In other embodiments, the injury is a fracture or weakened bone or periodontal disease.

[0166] In some embodiments, bone formation is systemic. Systemic bone formation refers to the formation of bone throughout the subject and can affect all of the bones within the subject's body. Subjects in need of systemic bone formation may be suffering from any of a variety of diseases or conditions. In some embodiments, the subject is suffering from low bone mass / density condition / disease (either primary or secondary), a fracture, a periodontal disease / condition, or a disease / condition that causes insufficient bone mineralization (e.g., osteogenesis imperfecta or HPP). Low bone mass can be determined by a variety of methods known to those of skill in the art. For example, low bone mass / density can be characterized by a T-score of less than about -0.5. Diseases / conditions of low bone mass / density include, but are not limited to, osteoporosis, osteopenia, and osteopetrosis-pseudoglioma syndrome (OPPG), glucocorticoid-induced low bone mass / density, osteogenesis imperfecta. In some other embodiments, the low bone mass condition / disease can be osteopenia or osteopetrosis-pseudoglioma syndrome (OPPG), HPP, or glucocorticoid-induced low bone mass / density or other diseases that result in secondary low bone density condition.

[0167] Local and / or systemic bone formation using the compounds or compositions of the present invention can be achieved according to any of a variety of methods. Methods of formulating and administering the compounds and compositions of the present invention (e.g., compounds or compositions of Formula I) are described in Section VII below. In some embodiments, a method of promoting bone formation includes implanting a medical device as described herein (e.g., in Section VIII below) into a subject in need thereof.

[0168] Methods that promote mineral deposition in osteoblasts and ultimately increase bone mineralization or density can be used in the treatment of diseases characterized by secondary osteoporosis (low bone mass). Such diseases include osteomalacia, polyostotic fibrous dysplasia, osteogenesis imperfecta, Paget's disease, rheumatoid arthritis, microgravity, osteoarthritis, prolonged inactivity or immobility, joint immobilization, osteomyelitis, celiac disease, Crohn's disease, ulcerative colitis, inflammatory bowel disease, gastrectomy, secondary osteoporosis, amenorrhea, Cushing's disease, Cushing's syndrome, type 2 diabetes, diabetes, eating disorders, hyperparathyroidism, hyperthyroidism, hypophosphatasia (HPP), hyperprolactinemia, Klinefelter syndrome, thyroid disorders, Turner syndrome, steroid-induced osteoporosis, seizure- or depression-induced osteoporosis, immobility, arthritis, cancer-induced secondary osteoporosis, gonadotropin-releasing hormone agonist-induced low bone mass, thyroid medication-induced low bone mass, Dilantin (phenytoin)-induced low bone mass, Depakote-induced low bone mass, chemotherapy-induced low bone mass, immunosuppressant-induced low bone mass, anticoagulant-induced low bone mass, Graves' disease, juvenile rheumatoid arthritis, malabsorption syndrome, anorexia nervosa, kidney diseases, anti-epileptic drug treatment (e.g., for epilepsy), corticosteroid treatment (e.g., for rheumatoid arthritis, asthma), immunosuppressive treatment (e.g., for cancer), inadequate nutrition (especially calcium, vitamin D), excessive exercise leading to amenorrhea (absence of menstruation), smoking and alcohol abuse, pregnancy-related osteoporosis, copper deficiency, type 2 aminoaciduria, Werner syndrome, Hallermann-Streiff syndrome, juvenile cortical hyperostosis deformansjuvenilis), type 2 methylmalonic aciduria, cystathionine β-synthase deficiency, exemestane, hyper-IgE syndrome, hemochromatosis, Singleton-Merten syndrome, β-thalassemia (homozygous), reflex sympathetic dystrophy, sarcoidosis, Winchester syndrome, Hallermann-Streiff syndrome (HSS), cyproterone, glycerol kinase deficiency, Bonnet-Dechaume-Blanc syndrome, prednisolone, heparin, cutis laxa osteodysplasia of the elderly, Torg osteolysis syndrome, orchidectomy, Fabry disease, pseudo-progeria syndrome, Wolcott-Rallison syndrome, ankylosing spondylitis, myeloma, systemic infantile hyalinosis, Albright hereditary osteodystrophy, anorexia nervosa, autoimmune lymphoproliferative syndrome, Brown-Séquard syndrome, Diamond-Blackfan anemia, eating disorders, galactorrhea, hyperprolactinemia, ovarian dysgenesis, kidney diseases, Menkes disease, menopause disorders, neuritis, ovarian insufficiency due to FSH resistance, familial ovarian insufficiency, premature aging, primary biliary cirrhosis, prolactinoma, familial prolactinoma, renal osteodystrophy, ulcerative colitis, weight loss, Werner syndrome, bone tumors, bone cancer, brittle bone diseases, osteonecrosis, congenital osteogenesis imperfecta, late-onset osteogenesis imperfecta, osteogenesis imperfecta, glucocorticoid-induced osteoporosis / osteopenia, and periodontal diseases are included, but not limited to these. Those skilled in the art will understand that other types of conditions, diseases, and treatments can also lead to osteoporosis.

[0169] Bone formation can be measured according to any of a variety of methods known to those skilled in the art. Methods for measuring bone formation include, but are not limited to, μCT (micro-CT), dual-energy X-ray absorptiometry (bone density), ultrasound, QCT, SPA, DPA, DXR, SEXA, QUS, X-ray, performed visually during surgical procedures, alizarin red S, serum osteocalcin, serum alkaline phosphatase, serum bone Gla-protein (BGP), bone mineral content, ash weight of bone, serum calcium, serum phosphorus, tartrate markers, and serum IGF-1.

[0170] Using many indicators of bone formation, the amount of bone formation, including bone density, can be measured and / or quantified. In some embodiments, bone formation is indicated by a 0.1% increase in bone density. In other embodiments, bone growth is indicated by an increase of 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 14%, 16%, 18%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% or more in bone density. Bone density can be measured in various ways, such as T-score and Z-score. The Z-score is the number of standard deviations above or below the average for the patient's age and gender. The T-score is the number of standard deviations above or below the average of a healthy 30-year-old adult of the same sex as the patient. Low bone mass is characterized by a T-score of -1 to -2.5. Osteoporosis is characterized by a T-score of less than -2.5. An improvement in the T-score or Z-score indicates bone growth. Bone density can be measured at various locations in the skeleton, such as the spine and hip. Those skilled in the art will understand that other methods of determining bone density are useful in the present invention.

[0171] V. Methods for Treating Kidney Disorders In another aspect, the present invention provides a method for treating kidney disorders in a subject suffering from kidney disorders by administering a therapeutically effective amount of a compound of the present invention (e.g., a compound or composition of Formula I as described in Section III above).

[0172] Kidney disorders can be caused by various diseases known to those skilled in the art. In some embodiments, kidney disorders are caused by infection, radiation, toxins, dehydration, or trauma. Toxins that cause kidney disorders include, but are not limited to, chemicals, poisons, and chemotherapeutic agents. Those skilled in the art will understand that other causes of kidney disorders can be treated by the methods of the present invention.

[0173] The renal disorders treatable by the compounds of the present invention include acute renal failure. Acute renal failure is also known as acute renal insufficiency or acute kidney injury. Acute renal failure usually leads to the retention of nitrogenous (urea and creatinine) and non-nitrogenous waste products that are normally excreted by the kidneys. Depending on the severity and duration of the renal dysfunction, this accumulation is accompanied by metabolic disorders such as metabolic acidosis (acidification of the blood) and hyperkalemia (elevation of potassium levels), changes in fluid balance, and effects on other organ systems. Acute renal failure may be characterized by oliguria or anuria (decrease or cessation of urine production), although non-oliguric acute renal failure can also occur.

[0174] Subjects may be characterized by (1) risk of acute injury, (2) kidney injury causing harm, (3) acute renal failure, and (4) loss of renal function. The risk of acute kidney injury is characterized by a 1.5-fold increase in serum creatinine or urine output less than 0.5 mL / kg body weight in 6 hours. Injury occurs when serum creatinine doubles to 2.0-fold or urine production is less than 0.5 mL / kg in 12 hours. Renal failure occurs when serum creatinine triples or creatinine increases to >355 μM (>44-fold increase), or when urine output is less than 0.3 mL / kg in 24 hours. Loss of renal function occurs when the subject has persistent acute renal failure or complete loss of renal function for more than 4 weeks.

[0175] Renal biopsy can be performed in the setting of acute renal failure to provide a definitive diagnosis and, in some cases, an idea of the prognosis, as long as the cause is not clear and appropriate screening tests are not definitely negative.

[0176] The renal therapeutic agent of the present invention can be used for a subject who has suffered from renal impairment or is at risk of chronic renal failure. As used herein, when it is reasonably expected that a subject suffers from a progressive loss of renal function associated with a progressive loss of functioning nephron units, the subject is said to be at risk of having or having chronic renal failure, or the subject is at risk of the need for renal replacement therapy (i.e., chronic hemodialysis, continuous peritoneal dialysis, or kidney transplantation). Whether a particular subject has or is at risk of chronic renal failure is a determination that can be routinely made by a person skilled in the relevant medical or veterinary art. Subjects at risk of chronic renal failure or its risk, or at risk of the need for renal replacement therapy, include, but are not limited to: subjects who can be considered to have chronic renal failure, end-stage renal disease, chronic diabetic nephropathy, hypertensive nephrosclerosis, chronic glomerulonephritis, hereditary nephritis, and / or renal dysplasia; subjects having a biopsy showing glomerular hypertrophy, tubular hypertrophy, chronic glomerulosclerosis, renal cell carcinoma, and / or chronic tubulointerstitial sclerosis; subjects undergoing ultrasound, MRI, CAT scan, or other non-invasive examinations showing renal fibrosis; subjects having an abnormal number of casts in the urine sediment; subjects having a GFR that is chronically less than about 50% of the predicted GFR of the subject, more specifically about 40%, 30%, or 20% or less; human male subjects weighing at least about 50 kg and having a GFR that is chronically less than about 50 mL / min, more specifically about 40 mL / min, 30 mL / min, or 20 mL / min or less; human female subjects weighing at least about 40 kg and having a GFR that is chronically less than about 40 mL / min, more specifically about 30 mL / min, 20 mL / min, or 10 mL / min or less; subjects having a number of functioning nephron units that is less than about 50% of the number of functioning nephron units possessed by a healthy but otherwise similar subject, more specifically about 40%, 30%, or 20% or less; subjects having only one kidney; and subjects who are kidney transplant recipients.

[0177] VI. Treatment Methods for Diabetes The compounds and compositions of the present invention are also useful in the treatment of diabetes. Accordingly, some embodiments of the present invention provide a method for treating diabetes. This method includes administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention (e.g., a compound or composition of Formula I as described in Section III above).

[0178] Diabetes is a disease in which the body cannot produce insulin at all or sufficiently, causing an increase in blood glucose levels in the affected individual. Without being bound by a particular theory, the compounds and compositions of the present invention are thought to be useful in treating diabetes by regenerating pancreatic cells. In some embodiments, the compounds of the present invention are thought to induce the regeneration of β-cells in the pancreas. See, for example, Wang P. et al., Nat Med., 21(4):383-388 (2015).

[0179] The diabetes therapeutics of the present invention can be used in subjects with pancreatic injury, in a prediabetic state, or with diabetes. As used herein, a subject considered to have pancreatic injury is one in which the natural production of insulin has decreased, been impaired, or is absent. Whether a subject is considered prediabetic or diabetic is determined by many factors, including fasting blood glucose levels. A subject is considered prediabetic if their fasting blood glucose level exceeds 100 mg / dL. If a subject's fasting blood glucose level exceeds 125 mg / dL, the subject is considered diabetic.

[0180] Pancreatic injury can be caused by various diseases known to those skilled in the art. In some embodiments, pancreatic injury is caused by an infectious disease, an autoimmune disease, radiation, a toxin, or trauma. Toxins that cause pancreatic injury include, but are not limited to, chemicals, poisons, and chemotherapeutic agents. Those skilled in the art will understand that other causes of pancreatic injury can be treated by the methods of the present invention.

[0181] In some embodiments, the disease being treated is type 1 diabetes. In some embodiments, the disease being treated is type 2 diabetes.

[0182] The compounds of the present invention can be administered continuously or in combination with other therapeutically useful agents for the treatment of diabetes. In some embodiments, the other therapeutic agent is an anti-diabetic agent. Anti-diabetic agents include, but are not limited to, lipid-lowering / lipid-regulating agents, agents for treating diabetic complications, anti-obesity agents, antihypertensive agents, SGLT1 inhibitors, SGLT2 inhibitors, anti-hyperuricemic agents, and agents for treating chronic heart failure, atherosclerosis, or related disorders.

[0183] VII. Method for treating osteopenia In another aspect, the present invention provides a method for treating osteopenia in a subject suffering from osteopenia by administering a therapeutically effective amount of a compound of the present invention (e.g., a compound or composition of Formula I as described in Section III above).

[0184] As shown in Figure 1, the bone density of an individual can be explained by the net loss (bone resorption) and increase (bone formation) of bone mass. In an individual with osteopenia, the net bone resorption is greater than bone formation, causing a decrease in osteopenia. In certain embodiments of the present invention, it is believed that osteopenia can be treated by inhibiting or reducing bone resorption while stimulating or promoting bone formation.

[0185] Bone resorption inhibitors are compounds that slow down the process of bone resorption. Bone resorption inhibitors include, but are not limited to, RankL inhibitors, denosumab, Prolia, cathepsin K modulators, alendronic acid, Fosamax, selective estrogen receptor modulators (SERMs), calcium, estrogen, bisphosphonates, and calcitonin.

[0186] The compounds and compositions of the present invention treat osteopenia by promoting bone formation. When the compounds of the present invention are administered continuously or in combination with one or more bone resorption inhibitors, the rate of bone resorption is inhibited or reduced and the rate of bone formation is stimulated.

[0187] The compounds and compositions of the present invention, as well as the bone resorption inhibitors described herein, can be administered continuously or in combination. Details of combination therapies are described in Section IX.C below.

[0188] The compounds and compositions of the present invention treat bone loss by promoting bone formation. In some embodiments, when the compounds of the present invention are administered continuously with one or more bone resorption inhibitors, the rate of bone resorption is inhibited or reduced and the amount of bone formation is maintained.

[0189] The compounds and compositions of the present invention are bone resorption inhibitors and can thus be administered to patients administered continuously, or patients can be administered one or more bone resorption inhibitors continuously, whereby the rate of bone resorption is inhibited or reduced and the amount of bone formation is maintained.

[0190] VIII. Cancer Therapies The compounds and compositions of the present invention are also useful in the treatment of cancer. Accordingly, some embodiments of the present invention provide methods of treating cancer. The method includes administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention (e.g., a compound or composition of Formula I as described in Section III above).

[0191] In some embodiments, the compounds of the present invention are useful in the treatment of cancer, proliferative disorders such as leukemia, and other disorders associated with uncontrolled cell proliferation such as psoriasis and restenosis. As defined herein, the anti-proliferative effects within the scope of the present invention can be demonstrated, for example, by using any of the A549, HT29, Saos-2, HeLa, or MCF-7 cell lines or by inhibiting cell proliferation in an in vitro whole cell assay by showing inhibition of a CDK enzyme (such as CDK2 or CDK4), MTT, or BRDU in a suitable assay. Such cell lines and enzyme assays can be used to determine whether a compound is anti-proliferative in light of the present invention.

[0192] As used herein, the term "cancer" includes, but is not limited to, the following cancers: breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, skin cancer, keratoacanthoma, lung cancer, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver and biliary tract cancer, kidney cancer, myelodysplastic disorders, lymphoid disorders, Hodgkin's disease, hairy cell leukemia, cancer of the buccal cavity and pharynx (oral), lip cancer, tongue cancer, oral cancer, pharyngeal cancer, small intestine cancer, colorectal cancer, colon cancer, rectal cancer, tumors of the brain and central nervous system, multiple myeloma, and leukemia. Those skilled in the art will understand that other cancers and proliferative disorders may be treated by the compounds and compositions of the present invention.

[0193] In some embodiments, the cancer is bone cancer, colon cancer, multiple myeloma, gastric cancer, colorectal cancer, prostate cancer, cervical cancer, lung cancer, pancreatic cancer, medulloblastoma, liver cancer, parathyroid cancer, endometrial cancer, or breast cancer. In some embodiments, the cancer is bone cancer. In some embodiments, the cancer is a cancer characterized by secondary low bone mass including, but not limited to, breast cancer and prostate cancer. In some embodiments, the cancer is a cancer that has metastasized to the bone.

[0194] IX. Formulation and Administration In some embodiments, the present invention provides a pharmaceutical composition comprising a compound described herein (e.g., a compound or composition of Formula I as described in Section III above) and a pharmaceutically acceptable excipient. In other embodiments, the composition further comprises a bone-conductive matrix.

[0195] The composition of the present invention can be in the form of a pharmaceutical composition comprising the antagonist and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include physiologically buffered saline solutions, other buffers, solvents, vehicles such as glycols and glycerols, oils such as olive oil, or aqueous solutions such as injectable organic esters. The choice of pharmaceutically acceptable carrier will depend in part on the chemical nature of the compound.

[0196] The compounds of the present invention can be formulated in a variety of different ways known to those skilled in the art. The pharmaceutically acceptable carrier is determined in part by the particular composition being administered and the particular method used to administer the composition. Thus, there are a variety of suitable formulations for the pharmaceutical compositions of the present invention (see, for example, Remington’s Pharmaceutical Sciences, 20 th ed., 2003, supra).

[0197] The pharmaceutically acceptable carrier can include, for example, a physiologically acceptable compound that acts to stabilize the composition or increase its absorption, or other excipients as needed. Physiologically acceptable compounds include, for example, carbohydrates such as glucose, sucrose, dextran, dextrin, cyclodextrin, or captisol, antioxidants such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins, or other stabilizers or excipients. Those skilled in the art will know that the choice of pharmaceutically acceptable carrier containing a physiologically acceptable compound is determined, for example, by the route of administration and its particular physicochemical properties.

[0198] Typically, such carriers must be non-toxic to the recipient at the dosages and concentrations used. Generally, the preparation of such compositions involves combining the therapeutic agent with buffers, antioxidants such as ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins, amino acids, glucose, maltose, sucrose, dextran, dextrin, cyclodextrin, or carbohydrates including captisol, chelating agents such as EDTA, glutathione, and other stabilizers and excipients. Neutral buffered saline or saline mixed with non-specific serum albumin are exemplary suitable diluents.

[0199] The amount of the compound or composition of the invention (e.g., a compound or composition of formula I described herein) to be administered to an individual will be determined, in part, by the degree of the disease and / or injury. Methods for determining the effective amount of a drug to be administered for diagnostic or therapeutic procedures are well known in the art and include Phase I, Phase II, and Phase III clinical trials, or pilot and pivotal trials (US Food and Drug Administration (FDA) device approval pathways). Typically, the drug is administered at a dose of about 0.0001 - 500 mg / kg body weight when administered systemically and at a concentration of about 0.1 nM - 1000 μM when administered directly to the site of injury.

[0200] The total amount of the compound or composition can be administered to the subject as a bolus, or by relatively short infusions, as a single administration, or using a divided treatment protocol in which multiple administrations are carried out over a longer period. One of ordinary skill in the art will know that the concentration of a particular compound or composition required to provide an effective amount in one or more regions of injury is determined by many factors including the age and general health of the subject and the route of administration, the number of administrations, and the nature of the compound. Taking these factors into account, one of ordinary skill in the art will adjust the specific dosage to obtain an effective amount for effectively promoting bone formation for therapeutic purposes.

[0201] The pharmaceutical preparation is preferably in unit dosage form. In such a form, the preparation is subdivided into unit doses containing an appropriate amount of the active ingredient. The unit dosage form can be a packaged preparation, and the package can include a packet of tablets, capsules, transdermal patches, ampoules, and individual amounts of the preparation such as powders in vials or ampoules or on osteoconductive matrices. Also, the unit dosage form can be a capsule, tablet, cachet, or troche itself, or an appropriate number of any of these in a packaged form. The composition can optionally also include other compatible therapeutic agents. Preferred pharmaceutical preparations can deliver the compounds of the present invention in sustained release formulations.

[0202] In some embodiments, the methods of the present invention include the application of the compounds described herein in cocktails containing other agents such as antibiotics, bactericides, bone anabolic agents, bone resorption inhibitors, and / or anti-inflammatory agents. Alternatively, the methods can include the sequential administration of the compounds described herein and one or more additional agents to an affected individual to optimize a treatment regimen. In such an optimized regimen, the agents containing the compounds of the present invention can be applied in any order and in any combination.

[0203] Individuals treated with the compounds and compositions of the present invention can be any mammal, such as a human or non-human mammal, such as a primate, dog, cat, horse, cow, goat, sheep, pig, mouse, or rat, or a commercially important animal or livestock.

[0204] In some embodiments, the individual being treated according to the methods of the invention is an individual who has been administered, or is being administered, an antiresorptive therapy. For example, in some embodiments, the antiresorptive therapy may be concurrent with the administration of the compounds or compositions of the invention. In some embodiments, the antiresorptive therapy and the treatment with the compounds or compositions of the invention are sequential (either an antiresorptive therapy preceding the treatment with the compounds or compositions of the invention, or a treatment with the compounds or compositions of the invention preceding the antiresorptive therapy). In some embodiments, the individual may have been previously treated with an antiresorptive agent. In some embodiments, the individual may be co-treated with an antiresorptive agent during the first part of the treatment course with the compounds or compositions of the invention, but the treatment with the antiresorptive agent may be discontinued during the second part of the treatment course. In some embodiments, the individual being treated according to the methods of the invention is not being treated with an antiresorptive agent. In some embodiments, the individual is treated with an antiresorptive agent after treatment with the compounds or compositions of the invention.

[0205] In some embodiments, the individual being treated according to the methods of the invention is an individual who has received, or is receiving, a combination of an antiresorptive agent and / or an anabolic bone therapy agent. For example, in some embodiments, the antiresorptive and / or anabolic bone therapy may be concurrent with the administration of the compounds or compositions of the invention. In some embodiments, the antiresorptive and / or anabolic bone therapy and treatment with the compounds or compositions of the invention are sequential administrations (either an antiresorptive therapy preceding treatment with the compounds or compositions of the invention, or treatment with the compounds or compositions of the invention preceding an antiresorptive therapy). In some embodiments, the individual may have been previously treated with an antiresorptive agent and / or an anabolic agent. In some embodiments, the individual may be treated concurrently with an antiresorptive agent and / or an anabolic agent during the first part of the treatment course with the compounds or compositions of the invention, but treatment with the antiresorptive agent and / or an anabolic agent may be discontinued during the second part of the treatment course. In some embodiments, the individual being treated according to the methods of the invention has not been treated with an antiresorptive agent and / or an anabolic agent. In some embodiments, the individual is treated with an antiresorptive agent and / or an anabolic agent after treatment with the compounds or compositions of the invention.

[0206] In some embodiments, the compounds and compositions of the invention are administered systemically. In some embodiments, the compounds and compositions of the invention are administered locally.

[0207] A. Systemic Delivery In some embodiments, the compounds and compositions of the invention are administered systemically. Systemic administration of the compounds and compositions of the invention can be used, for example, for the treatment of systemic diseases or conditions characterized by systemic effects, i.e., low bone mass (e.g., osteoporosis), diabetes, cancer, or renal disease.

[0208] The pharmaceutical composition of the present invention can be prepared for administration by various different routes. Usually, the type of carrier is selected based on the administration method. The pharmaceutical composition can be formulated for any suitable administration method including, for example, topical, oral, nasal, intrathecal, rectal, vaginal, sublingual, or parenteral administration, including injection or infusion subcutaneously, intravenously, intramuscularly, intra-sternally, transdermally, intracorporally, intrameatally, or intraurethrally. The pharmaceutical composition (for delivery, for example, by oral administration or injection) can be in the form of a liquid (such as an elixir, syrup, solution, emulsion, or suspension). The liquid pharmaceutical composition can include, for example, one or more of the following: water for injection, physiological saline, preferably physiological saline, Ringer's solution, isotonic sodium chloride, fixed oil that can serve as a solvent or suspension medium, polyethylene glycol, glycerin, propylene glycol, or other solvents; antibacterial agents; antioxidants; chelating agents; buffers such as acetate, citrate, phosphate; and agents for adjusting the osmotic pressure such as sodium chloride and dextrose. The parenteral preparation may be placed in an ampoule, disposable syringe, or multiple-dose vial made of glass or plastic. The use of physiological saline is preferred, and the injectable pharmaceutical composition is preferably sterile.

[0209] The formulations of the present invention are also suitable for administration to all body spaces / cavities including, but not limited to, the pleura, peritoneum, skull, mediastinum, pericardium, synovial sheath or bursa, epidural, intrathecal, intraocular, intra-articular, intra-disc, intramedullary, perispinal.

[0210] Formulations suitable for oral administration may be (a) liquid solutions such as an effective amount of a compound of the present invention suspended in a diluent such as water, physiological saline or PEG400, (b) capsules, sachets, depots, or tablets, each containing a predetermined amount of the active ingredient as a liquid, solid, granule, or gelatin, (c) suspensions in a suitable liquid, (d) suitable emulsions, and (e) patches. The dosage form may contain one or more of lactose, sucrose, mannitol, sorbitol, calcium phosphate, corn starch, potato starch, dextran, dextrin, cyclodextrin, captisol, microcrystalline cellulose, gelatin, colloidal silicon dioxide, talc, magnesium stearate, stearic acid, and other excipients, colorants, fillers, binders, diluents, buffers, wetting agents, preservatives, flavorings, dyes, disintegrants, pharmaceutically acceptable carriers, and other components classified as inactive ingredients by the FDA. The lozenge form may include lozenges containing the active ingredient in an inert base containing, in addition to the active ingredient and carriers known in the art, the active ingredient in a flavor, such as sucrose, and gelatin and glycerin or sucrose and acacia emulsion, gel, etc.

[0211] Specific oral formulations suitable for the present invention include, but are not limited to: buffered aqueous solutions with a pH of 4 to 10; non-buffered aqueous systems with a pH of 2 to 10; co-solvent aqueous solutions containing propylene glycol, glycerol, ethanol, or combinations thereof; aqueous solutions containing one or more emulsifiers such as one or more saturated polyglyceride saccharides (e.g., Gelucire®); aqueous suspensions containing methylcellulose (optionally containing sodium dodecyl sulfate, sodium lauryl sulfate, docusate, or polysorbate 80 at sub-critical micelle concentration (CMC)); aqueous solutions containing cyclodextrin (e.g., hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin); solutions containing one or more vegetable oils (e.g., safflower oil, soybean oil, oleic acid, etc.); non-aqueous solutions with or without emulsifiers such as PEG400 or 600, soybean oil / polysorbate 80 / sorbitan fatty acid ester (Span 80), mono / diglyceride or capric acid / caprylic acid (IMWITOR 742) / polysorbate 80 (70:30), polyethoxylated palm kernel oil / polyethylene glycol (PEG) 400 or 600 / water; surfactant aqueous solutions containing polysorbate 80 and SDS or SLS; oily suspensions containing soybean oil and safflower oil; and the preparation of the compounds or compositions of the present invention in nanoparticles.

[0212] Formulations suitable for intravenous bolus injection of a compound or composition include, but are not limited to: aqueous solutions containing buffered or unbuffered saline, optionally containing dextrose; co-solvent systems containing glycerin, ethanol, propylene glycol, PEG 300 or 400, glycofural, N-methylpyrrolidone (NMP), dimethylacetamide (DMA), dimethylformamide (DMF), dimethylisosorbide (DMI), dimethyl sulfoxide (DMSO), or combinations thereof in water; aqueous surfactant solutions containing polysorbate 80; aqueous solutions containing cyclodextrin (e.g., hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin); oily emulsions; plasma; and, optionally, aqueous suspensions containing methylcellulose and / or sodium dodecyl sulfate, sodium lauryl sulfate, docusate, or polysorbate 80 at sub-critical micelle concentration (CMC).

[0213] Formulations suitable for intravenous infusion of the compound or composition include, but are not limited to, aqueous solutions containing buffered or unbuffered saline, optionally containing dextrose, mannitol, or lactose, or any of the above formulations for intravenous bolus injection.

[0214] Formulations suitable for intramuscular, subcutaneous, or intraperitoneal administration include, but are not limited to: solutions in oil containing soybean oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, polysorbate 80, or sorbitan fatty acid esters; aqueous suspensions containing water, buffered or unbuffered saline, or dextrose (in water); or any of the above formulations for intravenous bolus injection.

[0215] Formulations suitable for intraocular administration include, but are not limited to: buffered or unbuffered aqueous solutions of pH 4 to 9 such as saline, optionally containing hydroxyethylcellulose; aqueous suspensions; or oily emulsions containing, for example, mineral oil, peanut oil, or petrolatum.

[0216] Typical formulations for topical / transdermal administration include creams, ointments, sprays, lotions, and patches. The topical / transdermal formulations of the present disclosure include propylene glycol, isopropyl myristate, PEG300, PEG400, petrolatum, or mixtures thereof, and may optionally include ethanol or isopropanol.

[0217] The compounds of the present invention may also be included in sustained-release formulations for long-term treatment after a single administration. In one embodiment, the formulation is prepared in the form of microspheres. The nanoparticles / microspheres can be prepared using the desired additional agents required for treatment as a homogeneous matrix of the compound containing a biodegradable controlled-release material. The nanoparticles / microspheres are preferably prepared in a size suitable for penetration and / or injection and are injected systemically or directly into the treatment site.

[0218] Some sustained-release embodiments include biodegradable and / or slowly dissolving polymeric substances. Such polymeric substances include polyvinylpyrrolidone, low molecular weight and medium molecular weight hydroxypropyl cellulose and hydroxypropyl methylcellulose, cross-linked sodium carboxymethyl cellulose, carboxymethyl starch, potassium methacrylate / divinylbenzene copolymer, polyvinyl alcohol, starch, starch derivatives, microcrystalline cellulose, ethyl cellulose, methyl cellulose, and cellulose derivatives, β-cyclodextrin, captisol, poly(methyl vinyl ether / maleic anhydride), glucan, scierozlucan, mannan, xanthan, alzinic acid and its derivatives, dextrin derivatives, glyceryl monostearate, semi-synthetic glycerides, glyceryl palmitostearate, glyceryl behenate, polyvinylpyrrolidone, gelatin, magnesium stearate, stearic acid, sodium stearate, talc, sodium benzoate, boric acid, and colloidal silica.

[0219] The sustained-release agent of the present invention may also contain adjuvants such as starch, pregelatinized starch, calcium phosphate, mannitol, lactose, sucrose, glucose, sorbitol, microcrystalline cellulose, gelatin, polyvinylpyrrolidone, methylcellulose, starch solution, ethylcellulose, gum arabic, tragacanth gum, magnesium stearate, stearic acid, colloidal silica, glyceryl monostearate, hydrogenated castor oil, wax, and mono-substituted, di-substituted, and tri-substituted glycerides. The sustained-release agent can also be prepared as commonly described in WO 94 / 06416 pamphlet.

[0220] B. Local delivery In some embodiments, the compounds and compositions of the present invention are administered locally. Local administration of the compounds and compositions of the present invention can be used, for example, for fracture healing, bone union (e.g., arthrodesis), orthopedic reconstruction, and periodontal repair. In some embodiments, local administration involves administering the compound or composition in combination with a suitable carrier material that can maintain the compound at the in vivo application site or provide a structural load. In some embodiments, the carrier is biocompatible, a matrix, biodegradable or absorbable in vivo, and / or porous enough to allow cell infiltration. In some embodiments, the compound or composition of the present invention (e.g., a compound or composition of formula I) is administered locally via an implantable (embedded) medical device.

[0221] The compounds and compositions of the present invention are useful in clinical applications in combination with a suitable delivery or support system (e.g., the scaffolds or matrices described herein). As disclosed herein, the matrix may be combined with a compound or composition of Formula I to reliably and reproducibly induce bone formation in a mammalian body. The matrix preferably comprises particles of a porous material. The pores are preferably of a size that allows for the migration of progenitor cells into the matrix and subsequent differentiation and proliferation. In some embodiments, the pore size of the matrix is at least 5 μm, for example, at least 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 250, 300, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1200, 1500, 1700, or 2000 μm. The matrix can be produced by tightly packing particulate material into a shape spanning a bone defect, or by structuring, as needed, a material that is biocompatible, preferably biodegradable or absorbable in vivo, and functions as a "temporary scaffold" and a substrate for the mobilization of migratory progenitor cells and as a substrate for subsequent anchorage and growth. In some embodiments, the scaffold or matrix consists of a mesh structure, a foam structure, a sponge structure, or a fibrous structure.

[0222] The scaffold or matrix for use in delivering the compounds of the present invention can include synthetic materials, biological materials, or combinations thereof. In some embodiments, the scaffold or matrix includes naturally occurring polymers, synthetic biodegradable polymers, synthetic non-biodegradable polymers, bioceramics, bioglasses, or combinations thereof. Natural polymers, synthetic polymers, bioceramics, and bioglasses for use in scaffolds are known in the art. See, for example, Dhandayuthapani et al., International Journal of Polymer Science, volume 2011, article ID 290602 (2011), which is incorporated herein by reference. Natural polymers include, but are not limited to, proteins (e.g., silk, collagen, gelatin, fibrinogen, elastin, keratin, actin, and myosin), polysaccharides (e.g., cellulose, amylose, dextran, chitin, chitosan, and glycosaminoglycan), and polynucleotides (e.g., DNA and RNA). Synthetic polymers include, but are not limited to, PLA, PGA, PLLA, PLGA, PCL, PLDLA, PDS, PGCL, PEA, PCA, PDLLA, PEU, and PBT. Bioceramics and bioglasses include, but are not limited to, HAP, TCP, CP ceramics, BCP, and TCP. In some embodiments, the scaffold or matrix is a hydrogel scaffold, a fibrous scaffold, a microsphere scaffold, a polymer-bioceramic composite scaffold, or a cell-free scaffold.

[0223] In some embodiments, the scaffold or matrix is a bone-conductive matrix. Non-limiting examples of suitable bone-conductive matrix materials include, for example: collagen; homopolymers or copolymers of glycolic acid, lactic acid, and butyric acid (including their derivatives); and ceramics, hydroxyapatite, tricalcium phosphate, biphasic calcium phosphate and other calcium phosphates, and calcium sulfate, or combinations thereof. Typically, the bone-conductive matrix contemplated herein includes at least one of the materials listed above. Other matrices useful in the present invention include, but are not limited to, biocomposite bone grafts, Kryptonite Bone Cement™ (Doctors Research Group, Oxford, Connecticut), Vitoss®, Vitoss® BA, Grafton® Orthoblend, Arthrex® allografts, cadaver bone, OSTEOSET®, NovaBone®, AUGMATRIX®, Mastergraft®, HydroSet®, PRO-DENSE™, PRO-STIM™, (porous) tantalum bone grafts, titanium mesh, titanium bone grafts, and genex® bone grafts. Combinations of these matrix materials may also be useful. The bone-conductive matrix can also include calcium salts, calcium sulfate, calcium phosphate, calcium phosphate cement, hydroxyapatite, coral-derived hydroxyapatite (HA), dicalcium phosphate, tricalcium phosphate (TCP), calcium carbonate, collagen, plaster of Paris, phosphopholine, borosilicate, bioactive glass, biocompatible ceramics, calcium phosphate ceramics, polytetrafluoroethylene, sulfates, collagen, and homopolymers or copolymers of glycolic acid, lactic acid, butyric acid (including their derivatives), and structural supports such as ceramics, hydroxyapatite, tricalcium phosphate, biphasic calcium phosphate and other calcium phosphates, and calcium sulfate.Other matrices useful in the present invention include, but are not limited to, biocomposite bone grafts, Kryptonite Bone Cement™ (Doctors Research Group, Oxford, Connecticut), Vitoss®, Vitoss® BA, Grafton® Orthoblend, Arthrex® allografts, cadaver bone, OSTEOSET®, NovaBone®, AUGMATRIX®, Mastergraft®, HydroSet®, PRO-DENSE™, PRO-STIM™, (porous) tantalum bone grafts, titanium mesh, titanium bone grafts, and genex® bone grafts or hydrogels.

[0224] In some embodiments, the osteoconductive matrix comprises an osteoinductive agent and, optionally, a structural support. The osteoinductive agent can be any agent that promotes bone formation. In some embodiments, the osteoinductive agent is an allograft, an autograft, demineralized bone, or periodontal ligament cells.

[0225] C. Combination Therapy In practicing the methods of the present invention, the pharmaceutical compositions can be used alone or in combination with other therapeutic or diagnostic agents. Further, the medical devices described herein include using the compounds of Formula I alone or in combination with other therapeutic or diagnostic agents. Additional drugs used in the combination protocols of the present invention may be administered separately or one or more of the drugs used in the combination protocol may be administered together, such as in a mixture. When administering one or more agents separately, the timing and schedule of administration of each agent can be different. The other therapeutic or diagnostic agents may be administered simultaneously with, separately from, or at different times than the compounds of the present invention.

[0226] In some embodiments, the compounds or compositions described herein (e.g., compounds or compositions of Formula I) are administered in combination with one or more other therapeutic agents. When the compounds of the invention are combined with another agent, the two can be administered simultaneously or separately. Simultaneous administration means within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours after administration of the compound of the invention, and within 1 to 7 days (e.g., 1, 2, 3, 4, 5, 6, or 7 days), within 1 to 4 weeks (e.g., 1, 2, 3, or 4 weeks), or within 1 to 18 months (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 months) of administering the other agent. Simultaneous administration also includes administering the other agent and the compound of the invention simultaneously, substantially simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other, or on the same day, in the same week, or in the same month), or in any order in sequence. In some embodiments, simultaneous administration involves administering another agent (e.g., an antiresorptive agent) for a period of time (e.g., several weeks, months, or years), then administering the compound or composition of Formula I for a period of time (e.g., several days, weeks, months, or years), and then administering the other agent (e.g., an antiresorptive agent) alone or in combination with the compound or composition of Formula I. In some embodiments, the other agents and compounds of the invention are each administered once, twice, three times, or more times a day, and can provide a preferred dosage level per day.

[0227] In some embodiments, simultaneous administration can be achieved by preparing a fixed-dose combination, i.e., a single pharmaceutical composition containing both the compound of the invention and a second therapeutic agent (e.g., an antiresorptive agent). In other embodiments, the compound of the invention and the second therapeutic agent are formulated separately.

[0228] One or more other therapeutic agents can be delivered by any suitable means. The pharmaceutical formulation is preferably in unit dosage form. In such form, the formulation is subdivided into unit doses containing an appropriate amount of the bone resorption inhibitor and / or the compound of the present invention. The unit dosage form can be a packaged formulation, and the package can include individual amounts of the formulation such as packeted tablets, capsules, and powders in vials or ampoules. Also, the unit dosage form can be a capsule, tablet, cachet, patch, or troche itself, or an appropriate number of any of these in a packaged form.

[0229] One or more other therapeutic agents may be present in any suitable amount and can be determined by various factors including, but not limited to, the weight and age of the subject, the state of the disease, etc. Suitable dosage ranges for one or more other therapeutic agents in combination with the compound or composition of the present invention include from about 0.0001 μg to about 10,000 mg, from about 0.0001 μg to about 1000 mg, from about 0.0001 μg to about 500 mg, from about 0.0001 μg to about 1000 μg, from 0.1 μg to about 10,000 mg, from about 0.1 μg to about 1000 mg, from about 0.1 μg to about 500 mg, from about 0.1 μg to about 1000 μg, from about 1 μg to about 1000 mg, from about 1 μg to about 500 mg, from about 1 μg to about 50 mg, from about 1 μg to about 1000 μg, from about 10 μg to about 1000 mg, from about 10 μg to about 500 mg, from about 10 μg to about 50 mg, from about 0.1 mg to about 10,000 mg, from about 1 mg to about 1000 mg, from about 10 mg to about 750 mg, from about 25 mg to about 500 mg, or from about 50 mg to about 250 mg. Suitable dosages for one or more other therapeutic agents in combination with the compound or composition of the present invention include about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1250, 1500, 1750, or 2000 mg.

[0230] One or more other therapeutic agents and the compounds or compositions of the present invention may be present in the compositions of the present invention in any suitable weight ratio, such as from about 1:100 to about 100:1 (w / w), or about 1:50 to about 50:1, about 1:25 to about 25:1, about 1:10 to about 10:1, about 1:5 to about 5:1 (w / w), or about 1:1 (w / w). Other dosages and dosage ratios of the bone resorption inhibitors and compounds of the present invention are suitable for the compositions and methods of the present invention.

[0231] The composition may also include other compatible therapeutic agents. The compounds described herein may be used in combination with each other, with other active agents, or with adjuvants that are not effective alone but may contribute to the effectiveness of the active agent.

[0232] In some embodiments, the compounds or compositions described herein (e.g., compounds or compositions of formula I) are administered to an individual being treated according to the methods of the present invention, in combination with or sequentially with, a bone resorption inhibitor. Bone resorption inhibitors include agents that retard or block bone resorption. Administration of the compounds or compositions described herein and the bone resorption inhibitor may promote local and / or systemic bone growth. In some embodiments, administration of the compounds or compositions described herein and the bone resorption inhibitor promotes systemic bone growth. Bone growth can be achieved by an increase in bone mineral content, an increase in bone density, and / or the growth of new bone. In other embodiments, local application of the compounds or compositions described herein and the bone resorption inhibitor achieves systemic bone growth.

[0233] Bone resorption inhibitors useful in the method of the present invention include, but are not limited to, denosumab, Prolia (registered trademark), RankL inhibitor, bisphosphonate (e.g., Fosamax (registered trademark), denosumab, Prolia (registered trademark), Actonel (registered trademark), Reclast (registered trademark), alendronate, Bonviva (trademark), Zometa (trademark), olpadronate, neridronate, Skelid (registered trademark), Bonefos (registered trademark)), selective estrogen receptor modulator (SERM) or analog (e.g., Evista (registered trademark)), calcitonin, calcitonin analog (e.g., Miacalcic (registered trademark)), parathyroid hormone, calcium degrading agent, calcium mimetic (e.g., cinacalcet), statin, anabolic steroid, lanthanum and strontium salts, and sodium fluoride, vitamin D or vitamin D analog, cathepsin K (CatK) inhibitor, prostaglandin inhibitor, or phosphodiesterase inhibitor (type E).

[0234] In some embodiments, the bone resorption inhibitor is denosumab.

[0235] The bisphosphonates useful in the method of the present invention can be any suitable bisphosphonates. In some embodiments, the bisphosphonates are nitrogen-containing compounds such as pamidronic acid (APD, Aredia®), neridronate, olpadronate, alendronic acid (Fosamax®), ibandronic acid (Bonviva™), risedronic acid (Actonel®), and zoledronic acid (Zometa™). In other embodiments, the bisphosphonates are non-nitrogen-containing compounds such as etidronic acid (Didronel®), clodronic acid (Bonefos®, Loron®), and tiludronic acid (Skelid®). Those skilled in the art will understand that other bisphosphonates are useful in the present invention.

[0236] The SERMs useful in the method of the present invention can be any suitable SERMs. In some embodiments, the SERMs can be clomiphene, raloxifene, tamoxifen, toremifene, bazedoxifene, lasofoxifene, or ormeloxifene. Those skilled in the art will understand that other SERMs are useful in the present invention.

[0237] The bone resorption inhibitor can also be any suitable calcitonin analog or cathepsin K inhibitor. In some embodiments, the calcitonin analogs useful in the method of the present invention include, but are not limited to, Miacalcic®. Those skilled in the art will understand that other calcitonin analogs are useful in the present invention.

[0238] The vitamin D analogs useful in the method of the present invention can be any suitable vitamin D analog. In some embodiments, vitamin D analogs useful in the method of the present invention include, but are not limited to, vitamin D1 (a molecular compound of ergocalciferol and lumisterol, 1:1), vitamin D2 (ergocalciferol or calciferol), vitamin D3 (cholecalciferol), vitamin D4 (22-dihydroergocalciferol), and vitamin D5 (sitocalciferol). Those skilled in the art will understand that other vitamin D analogs are useful in the present invention.

[0239] The RankL inhibitors useful in the present invention include any compound that inhibits RankL activity. For example, RankL inhibitors include, but are not limited to, denosumab or Prolia® (Prolia), a human monoclonal antibody. Those skilled in the art will understand that other RankL inhibitors are useful in the present invention.

[0240] In some embodiments, the individual being treated according to the method of the present invention is administered the compounds or compositions described herein (e.g., the compounds or compositions of Formula I) in combination with, or sequentially with, an osteoanabolic agent. Osteoanabolic agents include, but are not limited to, parathyroid hormone (PTH) or an analog thereof, a sclerostin inhibitor, a bone morphogenetic protein (BMP) or a BMP agonist, a population of bone marrow stem cells, or a population of mesenchymal stem cells.

[0241] In some embodiments, the bone anabolic agent is parathyroid hormone (PTH) or an analogue thereof (e.g., teriparatide (Forteo®)). In some embodiments, the bone anabolic agent is a sclerostin antibody (Mab) inhibitor. In some embodiments, the BMP is selected from the group consisting of BMP2, BMP7, and BMP4. In some embodiments, the BMP agonist is a compound described in Vrijens K, et al. PLoS One. 2013;8(3):e59045, the content of which is incorporated herein by reference. In some embodiments, the bone anabolic agent is a population of bone marrow stem cells. In some embodiments, the bone anabolic agent is a population of mesenchymal stem cells.

[0242] X. Medical Devices In some embodiments, the invention provides a medical device formed from a structural support, wherein the implantable portion of the structural support is adapted to be permanently implanted within a subject, the implantable portion adheres to bone, and the structural support carries at least a partial coating comprising the compound of claim 1 (comprising a compound of formula I as described herein (e.g., in section III above)). In some embodiments, the medical device is an orthopedic or periodontal medical device.

[0243] Another aspect of the invention is directed to medical implants. Such medical devices and implants include, for example, bone forming devices and methods of using the same for repairing endochondral bone and osteochondral defects, as taught in U.S. Patent Application Publication No. 20060177475 to David Rueger et al. (issued August 10, 2006), the subject matter of which is incorporated herein by reference, and as disclosed in U.S. Patent Nos. 6,190,880, 5,344,654, 5,324,819, 5,468,845, 6,949,251, 6,426,332, and 5,656,593, and U.S. Patent Application Publications Nos. 2002 / 0169122, 2002 / 0187104, 2006 / 0252724, and 2007 / 0172479.

[0244] These medical devices typically provide a structural support having a transplantable portion that is preferentially adapted to mechanically engage bone and / or cartilage, for example, as taught in U.S. Patent Application Publication No. 2006 / 0178752 to Joseph Vaccarino III, et al. (issued Aug. 10, 2006), the subject matter of which is incorporated herein by reference. These bone implants preferably contain an active agent in at least a portion thereof. As shown in U.S. Patent Application Publication No. 2006 / 0188542 to John Dennis Bobyn, et al. (issued Aug. 24, 2006), the subject matter of which is incorporated herein by reference, the active agent is preferably formulated to be locally deliverable to bone adjacent the implant in a sustained release or at least biphasic release scheme. In the latter case, a first phase rapidly releases a first amount of the active agent and a second and subsequent phases gradually release a second amount of the active agent, thereby regulating bone formation stimulated by the active agent.

[0245] Medical devices such as bone implants feature a transplantable portion having the compound or composition of the present invention (e.g., a compound or composition of Formula I) and promote more rapid and complete bone formation in situ. The transplantable portion of the medical device may desirably be at least partially or completely covered or impregnated with the compound or composition of the present invention. In some embodiments, the medical device is externally coated with the compound or composition described herein. In some embodiments, the external coating completely coats the transplantable portion of the structural support. In some embodiments, the structural support (e.g., a matrix or scaffold) contains the compound or composition described herein within the support, i.e., internally. In some embodiments, the structural support (e.g., a matrix or scaffold) includes an external coating of the compound or composition described herein and also contains the compound or composition within the support, i.e., internally.

[0246] The medical devices of the present invention include pins, rods, screws, plates, and orthopedic implants or dental implants. In some embodiments, the medical devices are made from materials including metals, polymers, ceramics, or combinations thereof. Metals useful for manufacturing the medical devices of the present invention include, but are not limited to, cobalt, chromium, chromium, stainless steel, titanium, titanium alloys, tantalum, Trabecular Metal (trademark). Polymers useful for manufacturing the medical devices of the present invention include, but are not limited to, ultra-high molecular weight polyethylene or high density polyethylene. In some embodiments, carbon fibers are combined with polyethylene. Additional useful polymers are described below. Ceramics useful for manufacturing the medical devices of the present invention include, but are not limited to, aluminum oxide, calcium phosphate, hydroxyapatite, zirconium oxide, silicon oxide.

[0247] In some other embodiments, the implantable portion of the structural support includes an osteoconductive matrix. The matrix material may promote bone growth. This may be desirable for materials such as teeth and bone grafts. Alternatively, if the implantable piece is load-bearing and formed, for example, of stainless steel, these implantable pieces are considered desirable when coated with the compounds or compositions of the present invention. In that case, it is also desirable to provide a separate matrix material that helps to form new bone growth.

[0248] In some embodiments, the matrix is composed of particles of a porous material. The pores are preferably of a size that allows for the migration of progenitor cells into the matrix and subsequent differentiation and proliferation. In some embodiments, the pore size of the matrix is at least 5 μm, for example, at least 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 110, 115, 120, 125, 150, 175, 200, 250, 300, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1250, 1500, 1750, or 2000 μm. In some embodiments, the scaffold or matrix consists of a mesh structure, a foam structure, a sponge structure, or a fibrous structure.

[0249] The scaffolds or matrices for use in the devices described herein may include synthetic and / or biological materials. In some embodiments, the scaffold or matrix includes naturally occurring polymers, synthetic biodegradable polymers, synthetic non-biodegradable polymers, bioceramics, bioglasses, bioactive glasses, biocomposites, or combinations thereof. Natural polymers and synthetic polymers, bioceramics, and bioglasses for use in scaffolds are known in the art. See, for example, Dhandayuthapani et al., International Journal of Polymer Science, volume 2011, article ID 290602 (2011), which is incorporated herein by reference. Natural polymers include, but are not limited to, proteins (e.g., silk, collagen, gelatin, fibrinogen, elastin, keratin, actin, and myosin), polysaccharides (e.g., cellulose, amylose, dextran, chitin, chitosan, and glycosaminoglycans), and polynucleotides (e.g., DNA and RNA). Synthetic polymers include, but are not limited to, PLA, PGA, PLLA, PLGA, PCL, PLDLA, PDS, PGCL, PEA, PCA, PDLLA, PEU, and PBT. Bioceramics and bioglasses include, but are not limited to, HAP, TCP, CP ceramics, BCP, and TCP. In some embodiments, the scaffold or matrix is a hydrogel scaffold, a fibrous scaffold, a microsphere scaffold, a polymer-bioceramic composite scaffold, or a cell-free scaffold.

[0250] In some embodiments, suitable matrices include composite biomaterials having a sponge-like structure, such as those containing phosphoholine and / or collagen, as taught by U.S. Patent Application Publication No. 2006 / 0188544 to Takashi Saito, published Aug. 24, 2006, which is incorporated herein by reference. Such coatings include, for example, monolayer and multilayer coatings as taught by U.S. Patent Application Publication No. 2006 / 0204542 to Zongtao Zhang et al., published Sep. 14, 2006, which is incorporated herein by reference, and those described in U.S. Pat. Nos. 6,949,251; 5,298,852; 5,939,039; and 7,189,263, and may be made by conventional methods including those taught therein.

[0251] In some embodiments, the matrix is an osteoconductive matrix. In some embodiments, the osteoconductive matrix includes an osteoinductive agent such as an allograft, autograft, demineralized bone or periodontal ligament cells, or combinations thereof. In some other embodiments, the osteoconductive matrix can be calcium salts, calcium sulfate, biphasic calcium phosphate, calcium phosphate, calcium phosphate cement, hydroxyapatite, coral-derived hydroxyapatite (HA), dicalcium phosphate, tricalcium phosphate (TCP), calcium carbonate, collagen, plaster of paris, phosphoholine, borosilicate, biocompatible ceramic, calcium phosphate ceramic, polytetrafluoroethylene, sulfate, borosilicate, bioactive glass, Mastergraft™ (Mastergraft) variants, Vitoss™ (Vitoss) variants, cement hydrogel, or combinations thereof. Those skilled in the art will understand that other osteoconductive matrices and osteoinductive agents are useful in the present invention.

[0252] In some embodiments, the medical devices described herein include both a compound of Formula I and an additional therapeutic agent. Suitable additional therapeutic agents include the combinations discussed in Section IX.C herein. For example, the medical device may include a compound of Formula I in combination with an osteoanabolic agent. In some embodiments, the medical devices described herein include a compound of Formula I in combination with a bone morphogenetic protein (BMP) or a BMP agonist. In some embodiments, the BMP is selected from the group consisting of BMP2, BMP7, BMP4. In some embodiments, the BMP agonist is a compound described in Vrijens K, et al. PLoS One. 2013;8(3):e59045, the content of which is incorporated herein by reference.

[0253] XI. Assays for Identifying Compounds for Treating Osteopenia Compounds useful in the methods of the present invention can be identified via a variety of methods known to those of skill in the art. Some exemplary methods for identifying such antagonists are described herein, including cell-based in vitro techniques (Journal of Bone and Mineral Research 2006, 21(11), 1738-1749). A general method for identifying a compound involves assessing the effect of an antagonist candidate on bone formation under controlled conditions. Preferably, bone formation is determined using dual energy x-ray absorptiometry (DEXA) on a living animal or microCT on an ex vivo sample. Preferred animals include rodents, and more preferably primates. The femur, tibia, and vertebrae are particularly useful subjects for such studies.

[0254] Briefly, the test animals are treated with a predetermined dose of the candidate compound. The control animals are treated with a control solution, preferably a non-irritating buffer or other carrier. When the candidate compound is delivered with a carrier, the control solution is ideally a carrier that does not contain the candidate compound. Multiple administrations of the candidate compound can preferably be applied to the test animals according to a predetermined dosing schedule. The dosing schedule can be, for example, over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 days or more, over several weeks, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 weeks or more, or over other periods of several months, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 months or more.

[0255] In an exemplary embodiment, local in situ administration of the candidate compound may be performed on the test animals, and the control animals are given an equal amount of the control solution without the candidate compound. The appropriate dosage will depend on the nature of the particular candidate compound being tested. Note that as an example, systemic administration (e.g., by oral or injection, e.g., intravenous, subcutaneous, or intramuscular) may also be used in dosing. The dosing performed by aerosol inhalation, eye drops, or oral ingestion must be in an amount sufficient to produce blood levels of the candidate compound similar to those achieved using systemic injection. The amount of the candidate compound that can be delivered by aerosol inhalation, eye drops, or oral ingestion to achieve these levels depends on the nature of the inhibitor used and can be determined by routine experimentation.

[0256] Once the dosing schedule is complete, both the test animals and the control animals are examined to determine the amount of bone formation present. This can be accomplished by any suitable method, but it is preferred to perform on live animals to analyze bone mineral content. Methods for micro-CT examination of animal bone are well known in the art. Candidate compounds suitable for use in promoting bone formation are identified by noting a significant increase in bone formation in the test animals as compared to the control animals. In some embodiments, the amount of bone formation in the test bone of the test animals is at least 0.5%, 1%, 3%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 22%, 24%, 26%, 28%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% or more compared to the equivalent bone of the control animals, the candidate compound is identified as being suitable for use in promoting bone formation. In some embodiments, compared to the control animals, bone formation increases by at least 3%, at least 5%, at least 7%, at least 10%, at least 12%, at least 14%, at least 16%, at least 18%, at least 20%, at least 30%, at least 40%, or at least 50% or more. Optionally, the level of bone formation can be calculated by determining the amount of bone formation present in each animal. The calculation can be performed by constructing a three-dimensional image of the bone formation and calculating the volume from the image with the aid of, for example, histomorphometry.

[0257] Examples of the above molecular modeling systems are composed of the CHARMm program and the QUANTA program (Polygen Corporation, Waltham, Massachusetts). CHARMm performs energy minimization and molecular dynamics functions. QUANTA performs molecular structure construction, graphic modeling, and analysis. QUANTA enables interactive construction, modification, visualization, and analysis of molecular interactions.

[0258] The compounds may be identified using a process known as computer or molecular modeling. This enables visualization of the three-dimensional atomic structure of the selected molecule and rational design of new compounds that interact with the molecule. The three-dimensional structure is typically determined from data from X-ray crystallography or NMR imaging of the selected molecule. Molecular dynamics requires force field data. A computer graphics system predicts how the new compound will link to the target molecule and experimentally manipulates the structures of the compound and the target molecule to achieve complete binding specificity. Prediction of how the molecule-compound interaction will change when small changes are made to one or both requires a computer specialized for calculations and molecular mechanics software, usually combined with a user-friendly menu-based interface between the molecular design program and the user.

[0259] XII. Specific Embodiments of the Present Disclosure Embodiment 1. A compound according to the following formula I:

[0260]

Chemical formula

[0261] or a salt, hydrate, prodrug, or isomer thereof, wherein R 1a 、R 1b 、R 1c 、R 1d 、and R 2 are each independently selected from hydrogen (H), halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkyl-OH, -O-C 1-6 alkyl-OH, C 3-6 cycloalkyl-C 1-4 alkoxy, and hydroxyl (-OH), provided that R However, R1a , R 1b , R 1c , R 1d , and R 2 Among them, two or less are H, A is

[0262] [Chemical formula]

[0263] and is R N is selected from the group consisting of heterocyclyl and heteroaryl, The heterocyclyl moiety is selected from monocyclic, fused bicyclic, and bridged cyclic heterocyclyls. The monocyclic heterocyclyl contains 4 to 7 ring members. The fused bicyclic heterocyclyl and the bridged bicyclic heterocyclyl contain 7 to 10 ring members. Each heterocyclyl moiety has 1 to 3 heteroatoms as ring members selected from nitrogen (N), oxygen (O), and sulfur (S). Each heterocyclyl moiety contains at least one nitrogen atom as a ring member and, optionally, is substituted with 1 to 3 R 5 moieties, The heteroaryl moiety contains 5 to 10 ring members, at least one ring member is a nitrogen atom, and, optionally, is substituted with 1 to 3 R 5 moieties, and R 5 are each selected from the group consisting of a hydroxyl group (-OH), C 1-3 alkyl, C 1-3 alkyl-OH, -O-C 1-3 alkyl, C 3-4 heteroalkyl, C 1-3 haloalkyl, -O-C 1-3 haloalkyl, halogen, and oxo, a compound.

[0264] Embodiment 2. R 1a , R 1b , R 1c , and R 1d are each hydrogen (H), halogen, C 1-6 alkyl, C1-6 Haloalkyl, C 1-6 Alkoxy, and C 1-6 Independently selected from the group consisting of haloalkoxy, Provided that R 1a , R 1b , R 1c , and R 1d Among them, two or less are H, the compound according to Embodiment 1. Embodiment 3. R 1a , R 1b , R 1c , and R 1d Each is independently selected from the group consisting of hydrogen (H), halogen, and C 1-6 Alkoxy, Provided that R 1a , R 1b , R 1c , and R 1d Among them, two or less are H, the compound according to Embodiment 2. Embodiment 4. R 1a , R 1b , R 1c , and R 1d Each is independently selected from the group consisting of hydrogen (H), fluorine (F), and methoxy, Provided that R 1a , R 1b , R 1c , and R 1d Among them, two or less are H, the compound according to Embodiment 3.

[0265] Embodiment 5. R 1c Is hydrogen (H) or fluorine (F), the compound according to any one of Embodiments 1 to 4.

[0266] Embodiment 6. R 1d Is hydrogen (H), the compound according to any one of Embodiments 1 to 5.

[0267] Embodiment 7. R 1b Is C 1-6The compound according to any one of Embodiments 1 to 6, which is an alkoxy group.

[0268] Embodiment 8. R 1b is the compound according to Embodiment 7, which is methoxy.

[0269] Embodiment 9. R 1a is the compound according to any one of Embodiments 1 to 8, which is a halogen.

[0270] Embodiment 10. R 1a is the compound according to Embodiment 9, which is fluorine (F).

[0271] Embodiment 11. R 2 is H, C 1-6 alkyl, and C 1-6 haloalkyl, and is the compound according to any one of Embodiments 1 to 10, which is selected from the group consisting of.

[0272] Embodiment 12. R 2 is C 1-6 haloalkyl, and is the compound according to Embodiment 11.

[0273] Embodiment 13. R 2 is CF3, and is the compound according to Embodiment 12.

[0274] Embodiment 14. R 2 is C 1-6 alkyl, and is the compound according to Embodiment 11.

[0275] Embodiment 15. R 2 is CH3, and is the compound according to Embodiment 14.

[0276] Embodiment 16. R NThe compound according to any one of Embodiments 1 to 15, which is heterocyclic.

[0277] Embodiment 17. R N is

[0278]

Chemical formula

[0279] The compound according to Embodiment 16, which is selected from the group consisting of

[0280] Embodiment 18. R N is

[0281]

Chemical formula

[0282] The compound according to Embodiment 17, which is selected from the group consisting of

[0283] Embodiment 19. R N is

[0284]

Chemical formula

[0285] The compound according to Embodiment 18, which is selected from the group consisting of

[0286] Embodiment 20. R N is

[0287]

Chemical formula

[0288] The compound according to Embodiment 19, which is

[0289] Embodiment 21. R N is the compound according to Embodiment 16, which is a monocyclic heterocyclyl.

[0290] Embodiment 22. The monocyclic heterocyclyl is each selected from the group consisting of piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl, and azetidinyl, optionally substituted with 1 to 2 R 5 moieties, the compound according to Embodiment 21.

[0291] Embodiment 23. R N is the compound according to Embodiment 16, which is a fused bicyclic or bridged heterocyclyl.

[0292] Embodiment 24. The fused bicyclic or bridged heterocyclyl is each optionally substituted with 1 to 2 R 5 moieties,

[0293]

Chemical Structure

[0294] selected from the group consisting of, the compound according to Embodiment 23.

[0295] Embodiment 25. R 1a and R 1b are each independently selected from the group consisting of hydrogen (H), halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, and C 1-6 haloalkoxy, R 1c and R 1d are independently H or halogen, provided that R 1a , R 1b , R 1c and R 1dTwo or less of them are H, R 2 is C 1-6 alkyl or C 1-6 haloalkyl, R N is a heterocyclyl optionally substituted with 1 to 3 R 5 substituents, and the heterocyclyl moiety is a monocyclic, bridged bicyclic, or fused bicyclic heterocycle, the compound according to Embodiment 1.

[0296] Embodiment 26. R 1a is halogen, R 1b is C 1-6 alkoxy, R 1c and R 1d are independently hydrogen (H) or fluorine (F), R 2 is C 1-6 haloalkyl, and R N is

[0297]

Chemical formula

[0298] selected from the group consisting of, the compound according to Embodiment 25.

[0299] Embodiment 27. R N is

[0300]

Chemical formula

[0301] selected from the group consisting of, the compound according to Embodiment 26.

[0302] Embodiment 28.

[0303]

Chemical formula

[0304] The compound according to Embodiment 1, selected from the group consisting of a salt, hydrate, or prodrug thereof.

[0305] Embodiment 29. The formate salt of the compound according to any one of Embodiments 1 to 28.

[0306] Embodiment 30. The sulfate salt of the compound according to any one of Embodiments 1 to 28.

[0307] Embodiment 31. The citrate salt of the compound according to any one of Embodiments 1 to 28.

[0308] Embodiment 32. The hydrochloride salt of the compound according to any one of Embodiments 1 to 28.

[0309] Embodiment 33. The prodrug of the compound according to any one of Embodiments 1 to 28.

[0310] Embodiment 34. A pharmaceutical composition comprising the compound according to any one of Embodiments 1 to 3 and a pharmaceutically acceptable excipient.

[0311] Embodiment 35. A method for promoting bone formation in a subject in need thereof, comprising administering a therapeutically effective amount of the compound according to any one of Embodiments 1 to 34 to the subject, thereby promoting bone formation in the subject.

[0312] Embodiment 36. The method according to Embodiment 35, wherein the bone formation is promoted at the surgical site of an injury or a local condition.

[0313] Embodiment 37. The method according to embodiment 36, wherein the bone formation is promoted at a surgical site selected from the group consisting of a fracture and weakened bone.

[0314] Embodiment 38. The method according to embodiment 36, wherein the subject requires spinal fusion, arthrodesis, or orthopedic or periodontal synthetic bone grafts or implants.

[0315] Embodiment 39. The method according to embodiment 35, wherein the bone formation is systemic.

[0316] Embodiment 40. The method according to any one of embodiments 35 to 39, wherein the subject has low bone mass / density, a fracture, or periodontal disease.

[0317] Embodiment 41. The method according to embodiment 40, wherein the low bone mass state is selected from the group consisting of osteoporosis, osteopenia, osteogenesis imperfecta (OI), osteoporosis-pseudoglioma syndrome (OPPG), and secondary low bone mass state.

[0318] Embodiment 42. The method according to embodiment 41, wherein the low bone mass state is selected from the group consisting of osteoporosis, osteopenia, and osteoporosis-pseudoglioma syndrome (OPPG).

[0319] Embodiment 43. The method according to any one of embodiments 35 to 42, further comprising administering a bone-conductive matrix to the subject.

[0320] Embodiment 44. The method according to embodiment 43, wherein the bone-conductive matrix comprises an osteoinductive agent selected from the group consisting of bone allografts, bone autografts, and periodontal ligament cells.

[0321] Embodiment 45. The method according to embodiment 43, wherein the bone-conductive matrix comprises a calcium salt, calcium sulfate, calcium phosphate, calcium phosphate cement, hydroxyapatite, coral-derived hydroxyapatite (HA), dicalcium phosphate, tricalcium phosphate (TCP), calcium carbonate, collagen, plaster of Paris, phosphoholine, borosilicate, biocompatible ceramic, calcium phosphate ceramic, demineralized bone matrix, biphasic calcium phosphate, biocomposite, tantalum, titanium, polytetrafluoroethylene, sulfate, hydrogel, bioactive glass, or a combination thereof.

[0322] Embodiment 45. The method according to any one of embodiments 35 to 45, wherein the compound is administered continuously or in combination with an antiresorptive agent.

[0323] Embodiment 46. The method according to embodiment 45, wherein the compound is administered to a patient who has been treated with the antiresorptive agent or who has previously been treated with an antiresorptive agent.

[0324] Embodiment 47. The method according to embodiment 45, wherein the antiresorptive agent is selected from the group consisting of denosumab, Prolia® (prolia), RankL inhibitor, bisphosphonate, selective estrogen receptor modulator (SERM), calcitonin, calcitonin analog, vitamin D, vitamin D analog, and cathepsin K inhibitor.

[0325] Embodiment 48. The method according to embodiment 45, wherein the antiresorptive agent is denosumab.

[0326] Embodiment 49. The method according to embodiment 45, wherein the antiresorptive agent is administered systemically.

[0327] Embodiment 50. The method according to embodiment 45, wherein the antiresorptive agent is administered locally.

[0328] Embodiment 51. The method according to any one of Embodiments 35 to 50, further comprising administering an osteoanabolic agent.

[0329] Embodiment 52. A medical device comprising a structural support, wherein the implantable portion of the structural support is adapted to be permanently implanted within a subject, the implantable portion adheres to bone, and the structural support carries at least a partial outer coating comprising the compound according to any one of Embodiments 1 to 34.

[0330] Embodiment 53. A method for treating bone loss in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of the compound according to any one of Embodiments 1 to 34 continuously or in combination with an antiresorptive agent to treat the bone loss in the subject.

Examples

[0331] XIII. Examples Example 1: 4-(2-(8-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine hydrochloride

[0332]

Chemical formula

[0333] To a dry tube were added 3-bromo-2-trifluoromethylpyridine (500 mg, 2.2 mmol), cesium carbonate (866 mg, 2.7 mmol), X-Phos (80 mg, 0.16 mmol), palladium(II) acetate (Pd(OAc)2) (25 mg, 0.1 mmol), and 2-fluoro-3-methoxy-6-chloroaniline (400 mg, 2.3 mmol). The mixture was diluted with toluene (5 mL), degassed, stirred at 120 °C overnight, then cooled to room temperature, diluted with ethyl acetate (EtOAc), washed with water and brine, dried over magnesium sulfate (MgSO4), and concentrated. The product was purified by silica chromatography using a gradient of 80% hexane / 20% EtOAc to 100% EtOAc to give N-(6-chloro-2-fluoro-3-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine (450 mg, 63%).

[0334] To a dry tube were added N-(6-chloro-2-fluoro-3-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine (450 mg, 1.4 mmol), potassium carbonate (390 mg, 2.8 mmol), and dimethylacetamide (DMA) (15 mL). The reaction mixture was degassed, and tert-butylphosphonium tetrafluoroborate ((t-Bu3)P HBF4) (80 mg, 0.3 mmol) and Pd(OAc)2 (30 mg, 0.15 mmol) were introduced. The reaction mixture was stirred at 120 °C overnight. An additional aliquot of the catalyst was added, and the reaction mixture was stirred at 120 °C overnight, then cooled to room temperature, diluted with EtOAc, washed with water and brine, dried over MgSO4, and concentrated. The product was purified by silica chromatography using a gradient of 85% hexane / 15% EtOAc to 50% hexane / 50% EtOAc. The product was re-purified by reverse-phase chromatography on a C 18 column using a gradient of 50% ACN / 50% water (0.1% FA) to 90% ACN / 10% water (0.1% FA). The dechlorinated material was still present. The product was further purified by HPLC C 18Further purification by column chromatography gave 8-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole (41 mg, 10%).

[0335] To a solution of dimethylformamide (DMF) (3 mL) containing 8-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole (41 mg, 0.15 mmol), 4-(2-chloroethyl)morpholine hydrochloride (54 mg, 0.3 mmol) was added, followed by sodium hydride (60% in oil, 35 mg, 0.9 mmol). The reaction mixture was stirred at 60 °C overnight, stirred at the lowest temperature of room temperature, quenched with sodium bicarbonate (NaHCO3) (saturated), diluted with EtOAc, then washed with NaHCO3 (saturated), brine, dried over MgSO4, and concentrated. The product was purified by silica chromatography using a gradient of 60% hexane / 40% EtOAc to 100% EtOAc to give 4-(2-(8-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine (29 mg, 51%). 1 H NMR (CDCl3, 400 MHz) δ 8.47 (d, 1H, J = 4.8 Hz), 8.02, (d, 1H, J = 5.2 Hz), 7.82 (d, 1H, J = 7.6 Hz), 7.03 (t, 1H, J = 7.2 Hz), 4.73 (t, 2H, J = 7.6 Hz), 4.03 (s, 3H), 3.58 (t, 4H, J = 4.8 Hz), 2.68 (t, 2H, J = 7.6 Hz), 2.47 (t, 4H, J = 4.8 Hz).

[0336] The product was diluted with dichloromethane (CH2Cl2), dioxane containing 1 mL of 4 M hydrogen chloride (HCl) was added, stirred for 10 minutes, and then evaporated to dryness. Trituration from CH2Cl2 / hexane gave the monohydrochloride salt. 11H NMR (DMSO-d6, 400 MHz) δ 11.31 (s, 1H), 8.53 (d, 1H, J = 4.8 Hz), 8.52 (d, 1H, J = 4.8 Hz), 8.20 (d, 1H, J = 8.8 Hz), 7.34 (t, 1H, J = 7.6 Hz), 4.95 (m, 2H), 4.02 (m, 2H), 3.99 (s, 3H), 3.81 (d, 2H), 3.53 (m, 2H), 3.39 (m, 2H), 3.19 (m, 2H). LCMS m / z 398.2 ([M+H] + , C 19 H 20 requires 398.2 for F4N3O2.)

[0337] Example 2: Synthesis of 4-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine hydrochloride

[0338]

Chemical Structure

[0339] In a Schlenk tube flame-dried under argon, 2 g of 2-bromo-4-fluoro-5-methoxyaniline (9.09 mmol, 1 eq) and 2.465 g of 3-bromo-2-(trifluoromethyl)pyridine were dissolved in 25 mL of dry degassed DMF. Subsequently, tris(dibenzylideneacetone)dipalladium(0) (Pd2dba3) (208 mg, 0.227 mmol, 2.5%) and 1,3-bis(2,6-diisopropylphenyl)imidazolium chloride (IPr·HCl) (193 mg, 0.455 mmol, 5%) were added. The resulting mixture was evacuated and refilled with argon (3 times), and then 10.91 mL of tetrahydrofuran (THF) containing 1.0 M potassium tert-butoxide (tBuOK) (10.91 mmol, 1.2 Eq) was added. The flask was evacuated and refilled with argon (3 times), and the reaction mixture was heated to 100 °C for 18 h. When the reaction was complete, water and EtOAc were added, and the resulting mixture was filtered through celite. The aqueous layer was extracted with EtOAc (×3), and the resulting organic layer was washed with brine, dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (CombiFlash® using 0% - 30% EtOAc / hexane over 20 min) to give 1.6 g of N-(2-bromo-4-fluoro-5-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine in 49% yield.

[0340] To a flame-dried Schlenk tube were loaded 1.67 g of N-(2-bromo-4-fluoro-5-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine (4.556 mmol, 1.0 Eq) and 1.260 g of potassium carbonate (K2CO3) (9.112 mmol, 2.0 Eq). Next, 40 mL of dry DMA was added, the flask was evacuated and refilled with argon three times, and then 103 mg of Pd(OAc)2 (0.457 mmol, 10 mol%) and 265 mg of tri-tert-butylphosphonium tetrafluoroborate (PtBu3.HBF4) (0.914 mmol, 20 mol%) were added. The flask was evacuated, refilled with argon three times, and the mixture was heated at 130 °C for 12 h. When the reaction was complete, water and EtOAc were added. The resulting mixture was filtered through celite, and the aqueous layer was extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (CombiFlash®, 0%–30% EtOAc / hexane) to give 776 mg of pure 6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole as a brown solid in 60% yield.

[0341] 4-(2-Chloroethyl)morpholine hydrochloride (105 mg, 0.562 mmol) was added to a stirred solution of the compound in DMF (3 mL) containing 6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole (80 mg, 0.281 mmol). Sodium hydride (NaH) (60% in oil, 67 mg, 1.7 mmol) was added in one portion, the flask was flushed with argon (Ar), and the mixture was heated at 60 °C overnight with stirring. The mixture was cooled to room temperature, diluted with EtOAc, washed once with water and once with brine, dried (MgSO4), and evaporated. After flash chromatography of the residue on silica (SiO2) (12 g) using 50%–100% EtOAc-hexane, preparative HPLC gave pure 4-(2-(6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine (32 mg, 29%). 1 1H NMR (CDCl3, 400 MHz) δ 8.47 (d, 1H, J = 4.8 Hz), 8.02 (d, 1H, J = 5.2 Hz), 7.77 (d, 1H, J = 10.4 Hz), 7.26 (d, 1H, J = 6.8 Hz), 4.75 (t, 1H, J = 8.0 Hz), 4.08 (s, 3H), 3.84 (t, 4H, J = 4.4 Hz), 2.91 (t, 2H, J = 8.0 Hz), 2.81 (t, 4H, J = 4.4 Hz).

[0342] HCl (2 M in Et2O, 0.36 mL, 0.72 mmol) was added via syringe to a stirred solution of 4-(2-(6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine (33 mg, 0.083 mmol) in CH2Cl2 (1 mL). Stirring was continued for 5 minutes, and then the volatiles were removed in vacuo. The residue was purified by trituration with 80% dichloromethane (CH2Cl2)-hexane to afford 4-(2-(6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine hydrochloride (36 mg, quantitative). 1 1H NMR (DMSO-d6, 400 MHz) δ 11.93 (bs, 1H), 8.47 (d, 1H, J = 5.2 Hz), 8.44 (d, 1H, J = 4.8 Hz), 8.30 (d, 1H, J = 11.2 Hz), 7.81 (d, 1H, J = 6.8 Hz), 5.00 (m, 1H), 4.10 (s, 3H), 4.01 (d, 2H, J = 12.8 Hz), 3.82 (t, 2H, J = 12.4 Hz), 3.60 (d, 2H, J = 12.4 Hz), 3.33 (m, 2H), 3.14 (m, 2H). LCMS m / z 398.2 ([M+H] + , C 19 H 20F4N3O2 requires 398.2.)

[0343] The compounds of Examples 3 to 13 are prepared by preparing a 6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole intermediate as described in Example 2. The final product of each example is prepared using an alkylation procedure similar to that described in Example 2. See Scheme 1.)

[0344] Example 3: 6-Fluoro-7-methoxy-9-(2-(4-methoxypiperidin-1-yl)ethyl)-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0345]

Chemical Structure

[0346] Free base: 1 H NMR (CD3OD, 400 MHz) δ 8.38 (d, 1H, J = 5.2 Hz), 7.93 (d, 1H, J = 4.8 Hz), 7.72 (d, 1H, J = 10.4 Hz), 7.08 (d, 1H, J = 7.2 Hz), 4.54 (t, 2H, J = 7.6 Hz), 3.99 (s, 3H), 3.28 (s, 3H) 3.18 (m, 1H), 2.77 (m, 2H), 2.65 (t, 2H, J = 7.6 Hz), 2.24 (t, 2H, J = 9.2 Hz), 1.85 (m, 2H), 1.57 (m, 2H).

[0347] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.45 (d, 1H, J = 4.8 Hz), 8.36 (d, 1H, J = 4.8 Hz), 8.07 (d, 1H, J = 10.4 Hz), 7.58 (d, 1H, J = 6.4 Hz), 5.03 (m, 2H), 4.15 (s, 3H), 3.81 (d, 0.74H, J = 12 Hz), 3.68 (m, 0.63H), 3.58 (m, 1.25H), 3.43 (m, 1.16H) 3.38 (s, 3H), 3.20 (m, 2H), 2.35 (m, 1.3H), 2.20 (m, 2H), 2.10 (m, 2H), 1.85 (m, 1.2H). LCMS m / z 426.2 ([M+H] + , C 21 H 24 requires 426.2 for F4N3O2.)

[0348] Example 4: 4-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)-1-methylpiperazin-2-one hydrochloride

[0349]

Chemical Structure

[0350] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.46 (d, 1H, J = 4.8 Hz), 8.00 (d, 1H, J = 4.8 Hz), 7.79 (d, 1H, J = 10.4 Hz),7.04 (d, 1H, J = 5.6 Hz), 4.61 (t, 2H, J = 7.2 Hz), 4.03 (s, 3H), 3.33 (m, 4H), 2.97 (S, 3H), 2.76 (m, 4H).

[0351] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.41 (d, 1H J = 4.8 Hz), 8.28 (d, 1H J = 4.8 Hz), 7.98 (d, 1H J = 10.8 Hz), 7.63 (d, 1H J = 6.8 Hz), 5.06 (t, 2H, J = 8.4 Hz), 4.13 (s, 3H), 4.09 (m, 2H), 3.55 (m, 4H), 3.55 (t, 2H, J = 8.0 Hz), 3.05 (s, 3H). LCMS m / z 425.2 ([M+H] + , C 20 H 21 requires 425.2 for F4N4O2.)

[0352] Example 5: 9-(2-(3-(Difluoromethoxy)azetidin-1-yl)ethyl)-6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0353]

Chemical Structure

[0354] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.39 (d, 1H, J = 4.8 Hz), 7.93 (d, 1H, J = 5.2 Hz), 7.72 (d, 1H, J = 10.0 Hz), 7.03 (d, 1H, J = 7.2 Hz), 6.12 (t, 1H, J = 73.6 Hz), 4.72 (m, 1H), 4.40 (t, 2H, J = 7.6 Hz), 4.00 (s, 3H), 3.66 (m, 2H), 3.14 (m, 2H), 2.82 (t, 2H, J = 7.2 Hz).

[0355] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.47 (d, 1H, J = 4.8 Hz), 8.40 (d, 1H, J = 4.0 Hz), 8.06 (d, 1H, J = 10.0 Hz), 7.56 (d, 1H, J = 7.2 Hz), 6.55 (t, 1H, J = 73.6 Hz), 4.91 (m, 1H), 4.75 (m, 2H), 4.46 (m, 2H), 4.20 (m, 2H), 4.15 (s, 3H), 3.72 (m, 2H). LCMS m / z 434.2 ([M+H] + , C 19 H 19 F6N3O2 requires 434.2.)

[0356] Example 6: 3-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)-8-oxa-3-azabicyclo[3.2.1]octane hydrochloride

[0357]

Chemical Structure

[0358] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.39 (d, 1H, J = 5.2 Hz), 7.93 (d, 1H, J = 4.8 Hz), 7.72 (d, 1H, J = 10.4 Hz), 6.94 (d, 1H, J = 6.8 Hz), 4.49 (t, 2H, J = 7.2 Hz), 4.20 (m, 2H), 3.99 (s, 3H), 2.62 (t, 2H, J = 7.6 Hz), 2.48 (d, 2H, J = 9.6 Hz), 2.40 (d, 2H, J = 9.2 Hz), 1.74 (m, 4H).

[0359] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.45 (d, 1H, J = 4.8 Hz), 8.35 (d, 1H, J = 4.8 Hz), 8.05 (d, 1H, J = 10.4 Hz), 7.83 (d, 1H, J = 6.4 Hz), 5.10 (m, 2H), 4.58 (m, 2H), 4.27 (s, 3H), 3.62 (d, 2H, J = 12.4 Hz), 3.42 (m, 2H), 3.25 (m, 2 H), 2.38 (m, 2H), 2.20 (m, 2H). LCMS m / z 424.2 ([M+H] + , C 21 H 22 F4N3O2 requires 424.2.)

[0360] Example 7: 8-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)-3-oxa-8-azabicyclo[3.2.1]octane hydrochloride

[0361]

Chemical Structure

[0362] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.45 (d, 1H, J = 5.2 Hz), 8.00 (d, 1H, J = 5.6 Hz), 7.78 (d, 1H, J = 10.4 Hz), 7.18 (d, 1H, J = 6.8 Hz), 4.59 (t, 2H, J = 7.2 Hz), 4.07 (s, 3H), 3.62 (d, 2H, J = 10.0 Hz), 3.49 (d, 2H, J = 10.0 Hz), 2.98 (m, 2H), 2.62 (t, 2H, J = 7.2 Hz), 1.84 (m, 4H).

[0363] Hydrochloride salt: 11H NMR (DMSO-d6, 400 MHz) δ 8.47 (d, 1H J = 4.8 Hz), 8.43 (d, 1H J = 4.8 Hz), 8.29 (d, 1H J = 11.2 Hz), 8.08 (d, 1H J = 7.2 Hz), 5.10 (t, 1H J = 8.0 Hz), 4.28 (d, 1H J = 12.0 Hz), 4.15 (m, 2H), 4.12 (s, 3H), 3.71 (d, 1H J = 12.0 Hz), 3.20 (m, 2H), 2.06 (bs, 4H). LCMS m / z 424.2 ([M+H] + , C 21 H 22 F4N3O2 requires 424.2.)

[0364] Example 8: 5-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)-2-oxa-5-azabicyclo[2.2.1]heptane hydrochloride

[0365]

Chemical Structure

[0366] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.45 (d, 1H, J = 5.2 Hz), 7.99 (d, 1H, J = 4.8 Hz), 7.77 (d, 1H, J = 10.4 Hz), 7.13 (d, 1H, J = 6.8 Hz), 4.53 (t, 2H, J = 7.6 Hz), 4.41 (bs, 1H), 4.05 (s, 3H), 3.91 (d, 1H, J = 8.0 Hz), 3.61 (d, 1H, J = 8.0 Hz), 3.46 (bs, 1H), 2.90 (m, 3H), 2.65 (d, 1H, J = 9.6 Hz), 2.05 (m, 2H). LCMS m / z 410.2 ([M+H] + , C20 H 20 F4N3O2 requires 410.2.)

[0367] Hydrochloride salt: 1 H NMR (CD3OD, 400 MHz) δ 8.44 (d, 1H J = 5.2 Hz), 8.32 (d, 1H J = 5.2 Hz), 8.05 (d, 1H J = 10.4 Hz), 7.58 (d, 1H J = 6.4 Hz), 5.01 (m, 2H), 4.23 (m, 1H), 4.15 (S, 3H), 3.90 (m, 2H), 3.65 (m, 2H), 3.45 (m, 1H), 3.18 (m, 1H), 2.30 (m, 3H). LCMS m / z 410.2 ([M+H] + , C 20 H 20 F4N3O2 requires 410.2.)

[0368] Example 9: 9-(2-(3,4-Dimethoxypyrrolidin-1-yl)ethyl)-6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0369]

Chemical formula

[0370] Free base: 1 H NMR (CDCl3, 400 MHz) δ 8.39 (d, 1H, J = 5.2 Hz), 7.93 (d, 1H, J = 5.2 Hz), 7.72 (d, 1H, J = 10.4 Hz), 7.03 (d, 1H, J = 6.8 Hz), 4.53 (t, 2H, J = 8.0 Hz), 3.98 (s, 3H), 3.82 (m, 2H), 3.36 (s, 6H), 2.92 (m, 2H), 2.82 (m, 4H).

[0371] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.42 (d, 1H J = 4.8 Hz), 8.29 (d, 1H J = 5.2 Hz), 8.01 (d, 1H J = 10.0 Hz), 7.55 (m, 1H), 4.96 (m, 2H), 4.28 (m, 2H), 4.14 (s, 3H), 3.88 (m, 2H), 3.57 (m, 2H), 3.49 (s, 6H), 3.35 (m, 2H). LCMS m / z 442.2 ([M+H] + , C 21 H 24 F4N3O2 requires 442.2.)

[0372] Example 10: 6-Fluoro-7-methoxy-9-(2-(3-methoxypyrrolidin-1-yl)ethyl)-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0373]

Chemical Structure

[0374] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.45 (d, 1H, J = 5.2 Hz), 7.99 (d, 1H, J = 4.8 Hz), 7.78 (d, 1H, J = 10.4 Hz), 7.12 (d, 1H, J = 6.8 Hz), 4.64 (t, 2H, J = 7.6 Hz), 4.04 (s, 3H), 3.95 (m, 1H), 3.31 (s, 3H), 3.97 (m, 1H), 2.85 (m, 3H), 2.66 (dd, 1H, J = 10.4, 6.0 Hz), 2.49 (q, 1H, J = 7.2 Hz), 2.13 (m, 1H), 1.88 (m, 1H).

[0375] Hydrochloride salt: 11H NMR (CD3OD, 400 MHz) δ 8.44 (d, 1H J = 4.8 Hz), 8.30 (d, 1H J = 5.6 Hz), 8.05 (d, 1H J = 10.4 Hz), 7.46 (bs, 1H), 4.95 (m, 2H), 4.22 (m, 1H), 4.13 (s, 3H), 3.92 (m, 2H), 3.56 (m, 2H), 3.38 (s, 3H), 3.25 (m, 2H), 2.42 (m, 1H), 2.22 (m, 1H). LCMS m / z 412.2 ([M+H] + , C 20 H 22 F4N3O2 requires 412.2.)

[0376] Example 11: 5-(2-(6-Fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)hexahydro-1H-furo[3,4-c]pyrrole hydrochloride

[0377]

Chemical Structure

[0378] Free base: 1 1H NMR (DMSO-d6, 400 MHz) δ 11.7 (bs, 1H), 11.2 (bs, 1H), 8.47 (d, 1H J = 4.8 Hz), 8.43 (d, 1H J = 4.8 Hz), 8.30 (d, 1H J = 10.8 Hz), 7.80 (dd, 1H J = 18.8, 6.8 Hz), 4.92 (m, 2H), 4.09 (s, 3H), 3.98 (m, 1H), 3.77 (m, 3H), 3.67 (m, 1H), 3.45 (m, 3H), 3.08 (m, 3H), 2.70 (m, 2H), 1.55 (m, 2H), 1.40 (m, 4 H). There are solvent and 5% impurities present.

[0379] Hydrochloride salt:1 1H NMR (DMSO-d6, 400 MHz) δ 12.2 (broad singlet, 1 H), 11.7 (broad singlet, 1H), 8.47, (doublet, 1H, J = 5.2 Hz), 8.44 (doublet, 1H, J = 5.2 Hz), 8.29 (doublet, 1H, J = 11.2 Hz), 7.97 (doublet, 0.5H, J = 6.4 Hz), 7.87 (doublet, 0.5H, J = 6.4 Hz), 5.00 (multiplet, 2H), 4.10 (singlet, 3H), 3.75 (multiplet, 4H), 3.40 (multiplet, 3H), 3.25 (multiplet, 1H), 3.08 (multiplet, 2H), 2.67 (multiplet, 1H), 1.65 (multiplet, 4H), 1.35 (multiplet, 8H). LCMS m / z 424.2 ([M+H] + , C 21 H 22 F4N3O2 requires 424.2. There are solvent and 5% impurities present.

[0380] Example 12: 9-(2-(4,4-Difluoropiperidin-1-yl)ethyl)-6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0381]

Chemical Structure

[0382] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.40 (doublet, 1H, J = 4.8 Hz), 7.92 (doublet, 1H, J = 4.8 Hz), 7.72 (doublet, 1H, J = 10.4 Hz), 7.00 (doublet, 1H, J = 6.8 Hz), 4.53 (triplet, 2H, J = 7.6 Hz), 3.99 (singlet, 3H), 2.71 (triplet, 2H, J = 7.6 Hz), 2.60 (triplet, 4H, J = 5.6 Hz), 1.95 (multiplet, 4H).

[0383] Hydrochloride salt:1 1H NMR (CD3OD, 400 MHz) δ 8.55 (broad doublet, 2H, J = 11.2 Hz), 8.11 (doublet, 1H, J = 10.0 Hz), 7.81 (doublet, 1H, J = 5.2 Hz), 5.16 (multiplet, 2H), 4.18 (singlet, 3H), 3.92 (multiplet, 2H), 3.50 (multiplet, 4H), 2.50 (multiplet, 4H). LCMS m / z 432.2 ([M+H] + , C 20 H 20 F6N3O requires 432.2.)

[0384] Example 13: 9-(2-(1H-1,2,4-triazol-1-yl)ethyl)-6-fluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole hydrochloride

[0385]

Chemical Structure

[0386] Free base: 1 1H NMR (CDCl3, 400 MHz) δ 8.45 (doublet, 1H, J = 5.2 Hz), 7.96 (doublet, 1H, J = 4.8 Hz), 7.94 (singlet, 1H), 7.67 (doublet, 1H, J = 10.0 Hz), 7.55 (singlet, 1H), 6.54 (doublet, 1H, J = 6.8 Hz), 4.91 (triplet, 2H, J = 5.6 Hz), 4.52 (triplet, 2H, J = 5.6 Hz), 3.88 (singlet, 3H).

[0387] Hydrochloride salt: 11H NMR (DMSO-d6, 400 MHz) δ 9.18 (s, 1H), 8.59 (s, 1H), 8.48 (d, 1H, J = 5.2 Hz), 8.43 (d, 1H, J = 5.2 Hz), 8.06 (d, 1H, J = 10.8 Hz), 7.10 (d, 1H, J = 6.8 Hz), 5.16 (t, 2H, J = 6.0 Hz), 4.87 (t, 2H, J = 6.0 Hz), 4.02 (s, 3H). LCMS m / z 380.1 ([M+H] + , C 17 H 14 requires 380.1 for F4N5O.)

[0388] Example 14: 4-(2-(6,8-Difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine hydrochloride

[0389]

Chemical Structure

[0390] A solution of N-chlorosuccinimide (839 mg, 6.28 mmol, 1 Eq) in 15 mL of acetonitrile was added dropwise at room temperature to a solution of 2,4-difluoro-3-methoxyaniline (1 g, 6.28 mmol, 1 Eq) in 35 mL of dry acetonitrile. The resulting mixture was heated at 60 °C for 12 h. When the reaction was complete, the solvent was evaporated in vacuo and the crude product was dissolved in ethyl acetate (100 mL). The organic layer was washed with water and then brine, dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (ethyl acetate:hexane = 1:9) to give 893 mg of 6-chloro-2,4-difluoro-3-methoxyaniline as a yellow oil in 73% yield.

[0391] In a flame-dried flask, 6-chloro-2,4-difluoro-3-methoxyaniline (404 mg, 2.08 mmol, 1.0 Eq) and 3-bromo-2-(trifluoromethyl)pyridine were dissolved in 10 mL of dry degassed xylene. Subsequently, cesium carbonate (CsCO3) (813 mg, 2.5 mmol, 1.2 Eq), Pd(OAc)2 (35 mg, 0.156 mmol, 7.5%), and XPhos (99 mg, 0.208 mmol, 10%) were added to the reaction mixture. The flask was evacuated and refilled with argon (×3), and the mixture was refluxed overnight. When the reaction was complete, water was added, and the product was extracted with EtOAc (×3). The combined organic layers were washed with brine, dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (CombiFlash® , isocratic, 5% EA / hexane) to give 223 mg of N-(6-chloro-2,4-difluoro-3-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine in 32% yield.

[0392] Pd(OAc)2 (30 mg, 0.132 mmol, 20%) 、 K2CO3 (182 mg, 1.32 mmol, 2.0 Eq), and tBu3P HBF4 (96 mg, 0.33 mmol, 50%) were loaded into a flame-dried flask. Dry degassed DMA (3 mL) containing N-(6-chloro-2,4-difluoro-3-methoxyphenyl)-2-(trifluoromethyl)pyridin-3-amine (223 mg, 0.658 mmol, 1.0 Eq) was added to the flask. The resulting mixture was heated at 165 °C overnight. The next day, water and EtOAc were added, and the mixture was filtered through celite. The product was extracted with EtOAc (×3), and the combined organic layers were washed with water (×2), brine (×2), dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (CombiFlash® , 5% EtOAc / hexane to 10% EtOAc / hexane) to give 53 mg of 6,8-difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole in 26.5% yield.

[0393] A stirred suspension of dry DMF (1 mL) containing 6,8-difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indole (28 mg, 0.093 mmol, 1.0 Eq), TBAI (6 mg, 0.02 mmol, 20%), and chloroethylmorpholine hydrochloride (95 mg, 0.186 mmol, 2.0 Eq) was added to NaH (60%) in oil (22 mg, 0.56 mmol, 6.0 Eq). The resulting mixture was heated at 60 °C overnight. Next, water was added and the compound was extracted with EtOAc (×3). The combined organic layers were washed with water (×3), brine (×1), dried over MgSO4, and concentrated in vacuo. The crude product was purified by flash chromatography (CombiFlash®, 0% - 40% EtOAc / hexane for 10 min, then 40% - 100% EtOAc / hexane for 5 min) to give 6 mg of 4-(2-(6,8-difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine in 15% yield. 1 H NMR (CDCl3, 400 MHz) δ 8.51 (d, 1H, J = 5.2 Hz), 8.02 (d, 1H, J = 5.2 Hz), 7.63 (dd, 1H, J = 9.2, 1.6 Hz), 4.74 (t, 2H, J = 7.2 Hz), 4.12 (s, 3H), 3.58 (t, 4H, J = 4.4 Hz), 2.68 (t, 2H, J = 7.2 Hz), 2.45 (t, 4H, J = 4.4 Hz). LCMS m / z 416.2 ([M+H] + , C 19 H 19 F5N3O2 requires 416.2.)

[0394] HCl (2 M in Et2O, 0.36 mL, 0.72 mmol) was added via syringe to a stirred solution of 4-(2-(6,8-difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine (0.083 mmol) in CH2Cl2 (1 mL). Stirring was continued for 5 minutes, and then the volatiles were removed in vacuo. The residue was purified by trituration with 80% dichloromethane (CH2Cl2)-hexane to afford 4-(2-(6,8-difluoro-7-methoxy-1-(trifluoromethyl)-9H-pyrido[3,4-b]indol-9-yl)ethyl)morpholine hydrochloride (quantitative). Hydrochloride salt: 1 H NMR (DMSO-d6, 400 MHz) δ 11.35 (s, 1H), 8.56 (s, 2H), 8.33 (d, 1H, J = 10.0 Hz), 4.96 (m, 2H), 4.06 (s, 3H), 4.98 (m, 2H), 4.06 (s, 3H), 3.98 (m, 2H), 3.82 (m, 2H), 3.52 (m, 2H), 3.38 (m, 2H), 3.18 (m, 2H).

[0395] Example 15: Modulation of Sclerostin / Wnt Activity The compounds synthesized according to the methods of Examples 1-14 were assayed for their ability to restore Wnt signaling in the presence of sclerostin, a known sclerostin antagonist, a monoclonal antibody to sclerostin that matches sclerostin. See Ellies et al., J Bone Miner Res 21:1738-1749 (2006). As shown in Table 1 below, sclerostin antagonized Wnt3a signaling in human embryonic cells. The addition of a known sclerostin antagonist inhibited the sclerostin inhibition of Wnt3a signaling, thereby restoring intracellular Wnt3a signaling (IC 100 )(data not shown). The compounds of Examples 1-14 also inhibited the sclerostin inhibition of Wnt3a signaling and restored intracellular Wnt3a signaling.

[0396] Example 16: Bone Formation Assay Mineralization (crystalline calcium phosphate formation) represents an in vitro model of bone formation. Using an assay that quantifies the amount of mineralization by measuring the total calcium after solubilization of the deposited crystalline calcium phosphate, sclerostin has previously been shown to inhibit mineralization of MC3T3-E1 (mouse calvarial) osteoblasts. Li et al., J Bone Miner Res 24:578-588 (2008). Compounds were assayed for their ability to rescue the inhibition of mineralization by sclerostin in MC3T3 osteoblasts according to the protocol described by Li et al. With sclerostin treatment alone, mineralization was significantly reduced when measured by calcium concentration (Table 1 and data not shown). The addition of the compounds of Examples 1 to 14 neutralized the inhibition of mineralization mediated by sclerostin, as reflected by the increase in calcium concentration.

[0397] Example 17: Metabolic Stability The compounds of the present invention (0.1 μM) were incubated with microsomes at 37 °C for a total of 60 minutes. The reaction contained human liver microsomal protein (0.1 mg / mL) pooled in potassium phosphate buffer containing NADPH. At indicated time points (0, 5, 15, 30, and 60 minutes). Samples were analyzed by LC / MS / MS and the remaining parent drug was calculated using Microsoft Excel (2007). Obach RS, Drug Metab Dispos. 27(11):1350-1359 (1999)).

[0398] [Table 1]

[0399] The foregoing invention has been described in some detail by way of illustration and example for the purposes of clarity of understanding, but those skilled in the art will appreciate that certain changes and modifications can be made within the scope of the appended claims. Further, each of the prior art documents provided herein is incorporated by reference in its entirety to the same extent as if each document were individually incorporated by reference.

Claims

1. A compound according to the following formula I: 【Chemical Formula 1】 Or a salt or hydrate thereof, wherein R 1a is fluorine (F); R 1b is methoxy; R 1c is hydrogen (H) or fluorine (F); R 1d is hydrogen (H); R 2 is hydrogen (H), C 1-6 alkyl, and C 1-6 haloalkyl, selected from the group consisting of, provided that no more than two of R1a, R1b, R1c, R1d, and R2 are H, A is 【Chemical Formula 2】 and R Ｎ is 【Chemical Formula 3】 A compound or a salt or hydrate thereof selected from the group consisting of.

2. R Ｎ is 【Chemical Formula 4】 The compound according to claim 1, selected from the group consisting of.

3. R Ｎ is 【Chemical Formula 5】 The compound according to claim 1.

4. 【Chemical Formula 6】 The compound according to claim 1, or a salt or hydrate thereof, selected from the group consisting of.

5. The formula: 【Chemical Formula 7】 The compound according to claim 1, or a salt thereof, having

6. Formula: 【Chemical Formula 8】 The compound according to claim 1, or a salt thereof, having

7. Formula: 【Chemical Formula 9】 The compound according to claim 1, or a salt thereof, having

8. The formate of the compound according to any one of claims 1 to 7.

9. The sulfate of the compound according to any one of claims 1 to 7.

10. The citrate of the compound according to any one of claims 1 to 7.

11. The hydrochloride of the compound according to any one of claims 1 to 7.

12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 11 and a pharmaceutically acceptable excipient.

13. A composition for promoting bone formation in a subject in need of bone formation, comprising a therapeutically effective amount of the compound according to any one of claims 1 to 11.

14. The composition according to claim 13, wherein the bone formation is promoted at a surgical site, an injury site or a site of a local condition.

15. The composition according to claim 14, wherein the bone formation is promoted at a surgical site selected from the group consisting of a fracture and weakened bone.

16. The composition according to claim 14, wherein the subject requires spinal fusion, arthrodesis, or synthetic bone grafts or implants for orthopedics or periodontics.

17. The composition according to claim 13, wherein the bone formation is systemic.

18. The composition according to claim 13, wherein the subject has low bone mass / density, a fracture, or periodontal disease.

19. The composition according to claim 18, wherein the low bone mass state is selected from the group consisting of osteoporosis, osteopenia, osteogenesis imperfecta (OI), osteoporosis-pseudoglioma syndrome (OPPG), and secondary low bone mass state.

20. The low bone mass state is selected from the group consisting of osteoporosis, osteopenia, and osteoporosis-pseudoglioma syndrome (OPPG ). The composition according to claim 19.

21. The composition according to any one of claims 14 to 20, further comprising an osteoconductive matrix.

22. The composition according to claim 21, wherein the osteoconductive matrix comprises an osteoinductive agent selected from the group consisting of bone allografts, bone autografts, and periodontal ligament cells.

23. The composition according to claim 21, wherein the osteoconductive matrix comprises calcium salts, calcium sulfate, calcium phosphate, calcium phosphate cement, hydroxyapatite, coral-derived hydroxyapatite (HA), dicalcium phosphate, tricalcium phosphate (TCP), calcium carbonate, collagen, plaster of paris, phosphopholine, borosilicate, biocompatible ceramics, calcium phosphate ceramics, demineralized bone matrix, biphasic calcium phosphate, biocomposites, tantalum, titanium, polytetrafluoroethylene, sulfates, hydrogels, bioglass, or combinations thereof.

24. whether the bone formation is promoted at the surgical site, or the subject has periodontal disease, osteoporosis, osteopenia, pseudoglioblastoma syndrome (OPPG), or rheumatoid arthritis, long-term immobility or immobilization arthropathy, osteomyelitis, celiac disease, Crohn's disease, ulcerative colitis, inflammatory bowel disease, gastrectomy, amenorrhea, Cushing's disease, Cushing's syndrome, eating disorder, hyperparathyroidism, hyperthyroidism, hyperprolactinemia, Klinefelter syndrome, thyroid disease, Turner syndrome, steroid-induced osteoporosis, seizure or depression-induced osteoporosis, immobility, arthritis, gonadotropin-releasing hormone agonist-induced low bone mass, thyroid medication-induced low bone mass, Dilantin (phenytoin)-induced low bone mass, Depakote-induced low bone mass, chemotherapy-induced low bone mass, immunosuppressant-induced low bone mass, anticoagulant-induced low bone mass, Graves' disease, juvenile rheumatoid arthritis, malabsorption syndrome, anorexia nervosa, kidney disease, antiepileptic drug treatment, corticosteroid treatment, immunosuppressive treatment, inadequate nutrition, smoking, alcohol abuse, pregnancy-related osteoporosis, copper deficiency, type 2 aminoaciduria, Werner syndrome, Hallermann-Streiff syndrome, juvenile cortical hyperostosis, type 2 methylmalonic aciduria, cystathionine beta synthase deficiency, exemestane, hyper IgE syndrome, hemochromatosis, Singleton-Merten syndrome, beta thalassemia, reflex sympathetic dystrophy, sarcoidosis, Winchester syndrome, Hallermann-Streiff syndrome (HSS), cyproterone, glycerol kinase deficiency, Bonnet-Dechaume-Blanc syndrome, prednisone use, heparin use, osteodystrophy cutis atrophicans senilis, Torg osteolysis syndrome, orchidectomy, Fabry disease, pseudoprogeria syndrome, Wolcott-Rallison syndrome, ankylosing spondylitis, multiple myeloma, generalized infantile hyalinosis, Albright hereditary osteodystrophy, anorexia nervosa, autoimmune lymphoproliferative syndrome, Brown-Séquard syndrome, Diamond-Blackfan anemia, galactorrhea, hyperprolactinemia, ovarian dysgenesis, kidney disease, Menkes disease, menopause, neuritis, ovarian insufficiency due to FSH resistance, familial ovarian insufficiency, premature aging, primary biliary cirrhosis, prolactinoma, familial prolactinoma, renal osteodystrophy, weight loss, Werner syndrome, bone cancer, brittle bone disease, osteonecrosis, osteomalacia, polyfibromatosis, osteogenesis imperfecta, Paget's disease,The composition according to claim 13, having a secondary low bone mass disease selected from the group consisting of osteoarthritis, osteomyelitis, late-onset osteogenesis imperfecta, and congenital osteogenesis imperfecta.

25. A medical device comprising a structural support, wherein the implantable portion of the structural support is adapted to be permanently implanted within a subject, the implantable portion adheres to bone, and the structural support carries at least a partial external coating, a partial internal coating, or both, at least one of which contains the compound according to any one of claims 1 to 11. The medical device.

26. The medical device according to claim 25, wherein osteoblasts / osteocytes can migrate to the structural support by the structural support and deposit bone mineral.

27. The medical device according to claim 25, wherein the structural support has at least a partial external coating, at least a partial internal coating, or at least a partial external and internal coating.

28. The medical device according to claim 25, wherein the structural support has at least a partial internal coating.

29. The medical device according to any one of claims 25 to 28, wherein the structural support is a cage, a pin, a rod, a screw, an orthopedic implant, or a dental implant.

30. The medical device according to claim 29, wherein the structural support contains a substance selected from metal, polymer, ceramic, or a combination thereof.

31. The medical device according to claim 29, wherein the structural support contains a polymer.

32. The medical device according to claim 30, wherein the metal contains stainless steel, cobalt, chromium, chrome, titanium alloy, titanium, tantalum, or trabecular metal (registered trademark). Claim 33 The medical device according to any one of claims 25 to 28, wherein the structural support is a titanium cage. Claim 34 The medical device according to claim 25, wherein the structural support is used for hip joint, knee joint, ankle joint, shoulder joint, dental implant or spinal fixation. Claim 35 The medical device according to claim 30, wherein the ceramic is calcium phosphate, aluminum oxide, zirconium oxide, silicon oxide or hydroxyapatite. Claim 36 The medical device according to claim 25, wherein the structural support is a scaffold or matrix containing synthetic material, biological material, biocomposite, naturally occurring polymer, synthetic biodegradable polymer, synthetic non-biodegradable polymer, bioceramic, bioglass or a combination thereof. Claim 37 The medical device according to claim 36, wherein the structural support is a scaffold or matrix containing a synthetic non-biodegradable polymer. Claim 38 The medical device according to claim 25, wherein the structural support is a scaffold or matrix containing silk, collagen, gelatin, fibrinogen, elastin, keratin, actin, myosin, cellulose, amylose, dextran, chitin, chitosan, glycosaminoglycan, DNA or RNA. Claim 39 The medical device according to claim 25, wherein the structural support is a scaffold or matrix containing PLA, PGA, PLLA, PLGA, PCL, PLDLA, PDS, PGCL, PEA, PCA, PDLLA, PEU, PBT, HAP, TCP, CP ceramic, BCP, or TCP. Claim 40 The medical device according to claim 25, wherein the structural support is a matrix containing particles of a porous material. Claim 41 The medical device according to claim 40, wherein the pore size of the matrix is at least 5 μm.

42. The medical device according to any one of claims 25 to 41, further comprising an osteoanabolic agent selected from the group consisting of parathyroid hormone (PTH) or an analog thereof, a sclerostin antibody inhibitor, a sclerostin inhibitor, BMP, a BMP agonist, a population of bone marrow stem cells, and a population of mesenchymal stem cells.

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