FAK activators for treating bone disorders

Abstract

A method to activate or enhance phosphorylation of focal adhesion kinase (FAK) are provided herein. Methods to treat musculoskeletal disease in a mammal, comprising the administration of one or more small molecules having FAK activation properties are also provided.

Description

FAK ACTIVATORS FOR TREATING BONE DISORDERSPRIORITY

[0001] This patent application claims the benefit of priority to United States Provisional Patent Application Serial No. 63 / 734,527, entitled “Catabolic Role of FAK-Activation in Osteoclastogenesis and Bone Resorption’’ filed on December 16, 2024, and to United States Provisional Patent Application Serial No. 63 / 750,596, entitled “Use of Small Molecule FAK Activators in Osteoclastogenesis and Bone Resorption” filed on lanuary 28, 2025 in the disclosure of which is incorporated herein in its entirety by reference.TECHNICAL FIELD

[0002] The present disclosure relates to devices and methods for treating bone disorders.BACKGROUND

[0003] Musculoskeletal diseases comprise a broad spectrum of conditions that affect the body's musculoskeletal system and its related components. These conditions can impact bones, joints, muscles, ligaments, and connective tissues, often causing pain, reduced mobility, and impaired function.SUMMARY

[0004] In some aspects, the techniques described herein relate to a molecule used to treat a musculoskeletal disease including a compound of Formula 1 or a salt thereof,2024-262-01r1(1), wherein

[0005] X1is -C-, -N-. or -O-; X2is -N- or -C-; and R1and R2are independently selected from the group consisting of -H, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.

[0006] In some aspects, the techniques described herein relate to a method to treat a musculoskeletal disease including administering to a subject in need thereof an effective amount of a compound of Formula 1 or a salt thereof.BRIEF DESCRIPTION OF DRAWINGS

[0007] The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments discussed in the present document.

[0008] FIG. 1 illustrates, by way of example, a graph showing the pharmacokinetics of a FAK activator.

[0009] FIGS. 2 A and 2B illustrate, by way of example, a series of graphs showing that effects of a FAK activator on cell viability and cell proliferation.

[0010] FIG. 3 illustrates, by way of example, osteoclast differentiation in culture.

[0011] FIG. 4A-4C illustrate, by way of example, a series of graphs showing the effects of a FAK activator on tartrate-resistant acid phosphatase (TRAP) activity, total number of osteoclasts, and relative TRAP activity with osteoclasts.2024-262-01

[0012] FIG. 5A-5C illustrate, by way of example, a series of graphs showing the effects of a FAK activator on osteoclasts nuclei distribution as determined by nuclei differential count analysis.

[0013] FIG. 6 A and 6B illustrate, by way of example, the effect of a FAK activator on osteoclasts in bone slices.

[0014] FIG. 7 A and 7B illustrate, by way of example, a series of graphs showing the effects of a FAK activator on osteoclasts in bone slices and on bone resorption.

[0015] FIG. 8 illustrates, by way of example, the effect of a FAK activator on osteoclasts in calvariae.

[0016] FIG. 9 illustrates, by way of example, the effect of a FAK activator on calvaria resorption.DETAILED DESCRIPTION

[0017] Focal Adhesion Kinase (FAK) is a cytoplasmic protein tyrosine kinase that can co-localize with integrins at sites of attachment to their ligands. In cells in culture, the sites of attachment can be manifested as regions of close contact with the underlying substrate called focal adhesions. Attachment of integrins to their extracellular matrix ligands can be a major regulatory stimulus for FAK, resulting in tyrosine phosphorylation and enzymatic activation. Other stimuli, e.g. growth factors, neuropeptides, cytokines, mechanical stimuli, can induce FAK phosphorylation / activation. FAK can associate with a large number of enzymes, adaptors, scaffold proteins, or a combination thereof. FAK can serve an enzymatic role, a scaffolding role, or both, in the transduction of signals.

[0018] A FAK can be organized into 4 domains including a Four-point-one, Ezrin, Radixin, Moesin, (FERM) domain, a catalytic domain, a prolinerich region, and a focal adhesion targeting (FAT) domain. The FERM domain can be located at the N-terminus of the FAK. The FERM domain can mediate a protein-protein interaction. The protein-protein interaction can be found in a cytoskeletal protein or a signaling protein. The catalytic domain can be located at the central domain of the FAK. The FAT domain can be located at the C-terminal 140 amino acids. The FAT domain can include a2024-262-01four a-helix bundle that can include binding sites for paxillin. The FAT domain can function to localize FAK to a focal adhesion. The proline-rich region can be located between the catalytic domain and the FAT domain. The proline-rich region can include two proline-rich regions that can serve as ligands for a SH3 domain of a signaling protein.

[0019] Upon activation, FAK autophosphorylates can create a docking site for a SH2 domain of a signaling molecule. The signaling molecule can include a Src family kinase, a phosphatidyl-inositol 3 ’-kinase, or both. The Src family kinase can promote phosphorylation of FAK on a tyrosine residue. The phosphorylation of the FAK can enable a maximal FAK catalytic activity, creation of an additional binding site for another protein, or both. FAK can associate with a tyrosine phosphorylated protein, including, but not limited to, pl30Cas, paxillin, or a combination thereof. The association can follow integrin dependent adhesion and phosphorylation of the tyrosine phosphorylated protein.

[0020] FAK can be a downstream signaling molecule that can function in the control of an integrin regulated biological process. The integrin regulated biological process can include, but is not limited to, cell migration, cell survival, cell proliferation, or a combination thereof. FAK can control tubule formation by endothelial cells or angiogenesis. FAK can function in the control of neurite outgrowth, netrin induced axonal guidance, or both.

[0021] Dysregulation of motility, survival and proliferation can be a hallmark of a human pathological condition including, but not limited to, cancer. Aberrant FAK signaling can result in an altered cellular phenotype, including, but not limited to, increased invasion, growth in soft agar, tumorigenicity, metastasis, or a combination thereof. FAK can be overexpressed in human cancer, suggesting that FAK can play a role in cancer pathology.

[0022] Pyk2 is a FAK-related kinase that can share the same overall domain structure and approximately 45% of the sequence identity of FAK. In contrast to FAK, which is ubiquitously expressed, Pyk2 can be more restricted in its expression in epithelial cells, hematopoietic cells, neural tissue, or a combination thereof. FAK can be more strongly activated by cell2024-262-01adhesion compared to Pyk2. Pyk2 can be more strongly activated by soluble ligands when compared to FAK. FAK and Pyk2 can have a number of common binding partners including, but not limited to, a Src family kinase, paxillin, or both. The number of common binding partners can indicate that FAK and Pyk2 can have a common signaling mechanism. Pyk2 can play an important role in macrophage function, osteoclast function, or both.

[0023] FAK can regulate a cytoskeletal dynamic for cell motility, fibroblast migration, or both. The regulation of the cytoskeletal dynamic can assist in wound healing. FAK can influence the proliferation of keratinocytes. The proliferation of keratinocytes can contribute to re-epitheli alization in a skin wound. FAK can modulate an inflammatory response. Modulation of the inflammatory response can contribute to the initial phase of wound healing. FAK can interact with a signaling molecule to balance pro-inflammatory and anti-inflammatory signals, aiding in a controlled repair process.

[0024] In the context of the gastrointestinal mucosa, FAK can contribute to an integrity of intestinal epithelial cells, a function of intestinal epithelial cells, or both. FAK can enable the maintenance of a barrier function. FAK can be involved in the response to injury, promoting cell migration, cell proliferation, or both, to facilitate mucosal healing.

[0025] In bone, FAK can enable a signal transduction pathway that can regulate mesenchymal stem cell lineage development in a skeletal system. Upon cell interaction with an extracellular signal from a cell-surface transmembrane receptor including, but not limited to, an integrins, a cytokines, a growth factor, a G protein-coupled receptor, or a combination thereof. FAK can activate a cellular process to mediate cell migration, survival, adhesion, proliferation, or a combination thereof. FAK activation and a FAK downstream signaling pathway can enable osteogenic differentiation of mesenchymal stem cells. An extracellular-integrin signal can be transmitted through FAK to activate osteoblast-related gene expression including, but not limited to, RUNX2 phosphorylation, osteogenesis, or both.2024-262-01

[0026] Osteoclasts are large multinucleated cells that can carry out bone resorption. The process of osteoclastogenesis can occur within a bone microenvironment through differentiation of hematopoietic stem cells into mature osteoclasts. Hematopoietic stem cells can differentiate into bone marrow macrophages when exposed to Macrophage-colony Stimulating Factor (M-CSF). Bone marrow macrophages can fuse with each other to form mature osteoclasts when exposed to Receptor Activator of Nuclear factor Kappa beta ligand (RANK-L). The RANK-L can bind to a Receptor Activator of Nuclear factor Kappa beta (RANK) to induce the activation of master transcription factor nuclear factor of activated T cells 1 (NFATcl). The activation of NFATcl can induce the expression of an osteoclast marker including, but not limited to, Tartrate-resistant Acid Phosphatase (TRAP), Cathepsin K (CTSK), Calcitonin Receptor (Calc R), Osteoclast-associated Receptor (OSCAR), osteoclast-specific transmembrane protein (OC-STAMP), or a combination thereof. Mature osteoclasts can migrate to a bone surface where the mature osteoclasts can form a sealing zone. The sealing zone can be acidified, pumped with various proteases, or both, to trigger bone resorption.

[0027] Inhibition of FAK activation can negatively regulate integrin-mediated osteoclastogenesis. In this disclosure, compound 30 (depicted by Formula 5, herein), a small molecule that specifically binds to and activates FAK, can used to probe the effects of FAK activation on osteoclast differentiation and function and bone resorption.

[0028] Drug-like small molecule FAK activators can be utilized as a method to target FAK activation in osteoclast differentiation and function. Drug-like small molecule FAK activators can be used to treat one or more conditions. For example, drug-like small molecule FAK activators can be a therapeutic approach for various bone-related conditions through the ability to target FAK activation in osteoclast differentiation and function. Drug-like small molecule FAK activators can be administered through one or more delivery methods, depending on the condition being treated. For systemic delivery applications, drug-like small molecule FAK activators can treat bone diseases characterized by increased bone mass or density including, but2024-262-01not limited to, generalized bone sclerosis, osteopetrosis, a genetic disorder associated with excessive bone formation, a genetic disorder associated with impaired bone resorption, an acquired disorder associated with excessive bone formation, an acquired disorder associated with impaired bone resorption, or a combination thereof. Drug-like small molecule FAK activators can be administered locally for treating one or more forms of heterotopic ossification, including, but not limited to, cases induced by traumatic brain injury, spinal cord injury, severe burns, or a combination thereof. Drug-like small molecule FAK activators can be beneficial in treating fibrodysplasia ossificans progressiva (FOP) through systemic delivery methods. The one or more therapeutic applications can leverage the ability of drug-like small molecule FAK activators to influence osteoclast activity, providing one or more treatment options for conditions characterized by abnormal bone formation or density

[0029] A genetic mutation can impact bone health through disrupted osteoclast function. In an example, a mutation in a gene including, but not limited to, TCIRG1, M-CSF, CLC-7, CTSK, or a combination thereof, can lead to osteopetrosis, a condition where bones become overly dense and prone to fractures. The mutation can interfere with the normal process of bone resorption, resulting in abnormal bone structure and potential complications throughout the skeletal system.

[0030] One example is pycnodysostosis, a rare genetic disorder caused by mutations in the Cathepsin K gene. Pycnodysostosis can manifest as short stature; distinctive features affecting the skull, clavicles, or digits; or a combination thereof. The underlying mechanism of pycnodysostosis can involve impaired osteoclast function, specifically in the ability of the osteoclast to break down Type I collagen in bone. While the osteoclasts can still process the mineralized portion of bone matrix, osteoclasts can struggle with organic component degradation, leading to a situation where the osteoclasts can include visible fibrillary collagen in intracellular vacuoles. Pycnodysostosis can be characterized by an increased number of osteoclasts, yet their functionality remains compromised, resulting in one or more2024-262-01distinctive bone abnormalities. Drug-like small molecule FAK activators can be used to treat pycnodysostosis.COMPOUNDS

[0031] Provided herein is the further development of the observation that certain small molecules or compounds that can mimic the tertiary structure of one subdomain of FAK can result in increased FAK activation. No currently available therapeutic specifically activates FAK, although numerous growth factors and cytokines are noted to activate FAK along with many other signals within the cell. No currently available agent can directly and specifically promote the induction of osteoclastogenesis and bone resorption.

[0032] Induction of osteoclastogenesis and bone resorption can play a role in the treatment of musculoskeletal disease. Musculoskeletal disease can encompass a diverse range of conditions affecting the skeletal system.Osteogenesis imperfecta, commonly known as brittle bone disease, is a genetic disorder that can cause a bone to break due to a collagen defect. Osteoporosis can represent a metabolic bone disease characterized by decreased bone mass, increased fracture risk, or both. Osteoporosis can be common in a postmenopausal woman, an elderly individual, or both. Paget's disease can cause abnormal bone remodeling, resulting in an enlarged bone, a misshapen bone, or both, that can be prone to a fracture. Multiple myeloma, a cancer of plasma cells, can cause a bone lesion, a weakening of the skeletal structure, or both. A bone infection like osteomyelitis can occur through various pathways, causing inflammation, destruction of bone tissue, or both. Fibrous dysplasia can involve replacement of normal bone tissue with fibrous connective tissue, leading to a weakened bone, a bone deformity, or both. Hyperparathyroidism can affect bone metabolism through excessive parathyroid hormone production, causing bone weakening, a calcium imbalance, or both. Rickets can result from a vitamin D deficiency or a vitamin D resistance. Rickets can lead to soft bones, deformed bones or both. Osteomalacia can cause bone softening due to impaired bone mineralization. Giant cell tumor of bone can cause bone2024-262-01destruction, joint dysfunction, or both. Bone metastases, from breast, prostate, and lung cancers, can lead to a pathological fracture, severe bone pain, or both. Osteopetrosis can lead to excessive bone density due to impaired osteoclast function. Primary hyperparathyroidism can lead to increased bone formation. Fibrous dysplasia can lead to replacement of bone with fibrous tissue, causing localized increased bone mass. Acromegaly can lead to excessive growth hormone causing increased bone mass in specific areas.

[0033] The molecules or compounds disclosed herein can exert their therapeutic effects by directly activating FAK. While tyrosine phosphatase inhibition can activate FAK, such an approach can non-selectively activate multiple other kinases. The selectivity of FAK activation can permit administration of high doses of the activator with reduced off-target toxicity.

[0034] Compounds of the following formulas are provided herein for treating musculoskeletal diseases as described above.whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -II, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.2024-262-01

[0035] In some aspects, the (Ci-C2o)hydrocarbyl is chosen from (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0036] In some aspects, the compound is represented by Formula 2:whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -H, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof. The (C1-C20) hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0037] In some aspects, the compound is represented by Formula 3:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

[0038] In some aspects, the compound is represented by Formula 4:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

[0039] In some aspects, the compound is represented by Formula 5:2024-262-01

[0040] In some aspects, the compound is represented by Formula 6:(6), wherein n is any positive integer.

[0041] In some aspects, the compound is represented by Formula 7:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

[0042] In some aspects, the compound is represented by Formula 8:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

[0043] Some aspects provide for combination therapy, including, but not limited to, administration of a FAK activation small molecule, administration of a FAK activation compound, immunotherapy, immunosuppressive therapy, anti-ulcer therapy, or a combination thereof. For example, oral administration of the FAK activator can be synergistic with an immunosuppressive therapy in inflammatory bowel disease, enhancing quality of life at a lower immunosuppressive dosing. Oral administration of the FAK activator can be through direct dosing or via an enteral release formulation.PHARMACEUTICAL COMPOSITIONS AND METHODS OF ADMINISTRATION

[0044] The present disclosure also contemplates pharmaceutical compositions comprising one or more compounds disclosed herein, one or more pharmaceutically acceptable carriers, diluents, excipients, or a combination thereof. A “pharmaceutical composition” can refer to a chemical or biological composition suitable for administration to a subject (e.g., mammal). Such compositions can be specifically formulated for administration via one or more of a number of routes including, but not limited to, buccal, cutaneous, epicutaneous, epidural, infusion, inhalation,2024-262-01intraarterial, intracardial, intracerebroventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, pulmonary, rectally via an enema or suppository, subcutaneous, subdermal, sublingual, transdermal, transmucosal, or a combination thereof. In addition, administration can be through capsule, drops, foams, gel, gum, injection, liquid, patch, pill, porous pouch, powder, tablet, other suitable means of administration, or a combination thereof. Additionally, the pharmaceutical composition can be topically or directly applied to a tissue.

[0045] A “pharmaceutical excipient” or a “pharmaceutically acceptable excipient” can include a carrier, such as a liquid, in which an active therapeutic agent can be formulated. The excipient generally does not provide any pharmacological activity to the formulation, though it can provide chemical stability, biological stability, a release characteristic, or a combination thereof. Examples of suitable formulations can be found, for example, in Remington, The Science and Practice of Pharmacy, 20th Edition, (Gennaro, A. R., Chief Editor), Philadelphia College of Pharmacy and Science, 2000, which is incorporated by reference in its entirety.

[0046] As used herein “pharmaceutically acceptable carrier” or “excipient” can include any and all solvents, a dispersion media, a coating, an antibacterial agent, an antifungal agent, an isotonic, an absorption delaying agent, or a combination thereof. The carrier can be suitable for, among other applications, parenteral administration. Alternatively, the carrier can be suitable for intravenous, intraperitoneal, intramuscular, sublingual, oral administration, or a combination thereof. Pharmaceutically acceptable carriers can include, but is not limited to, sterile aqueous solutions or dispersions, sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion, or a combination thereof. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.2024-262-01

[0047] Pharmaceutical compositions can be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable for high drug concentration. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

[0048] In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, monostearate salts and gelatin.Moreover, the compounds described herein can be formulated in a time release formulation, for example in a composition that includes a slow-release polymer. The active compounds can be prepared with carriers that will protect the compound against rapid release, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid and polylactic, polyglycolic copolymers (PLG). Many methods for the preparation of such formulations are known to those skilled in the art.

[0049] Oral forms of administration are also contemplated herein. The pharmaceutical compositions can be orally administered as a capsule (hard or soft), tablet (film coated, enteric coated or uncoated), powder or granules (coated or uncoated) or liquid (solution or suspension). The formulations can be conveniently prepared by any of the methods well-known in the art. The pharmaceutical compositions can include one or more suitable production aids or excipients including fillers, binders, disintegrants, lubricants, diluents, flow agents, buffering agents, moistening agents, preservatives, colorants, sweeteners, flavors, and pharmaceutically compatible carriers.2024-262-01

[0050] The compounds can be administered by a variety of dosage forms as known in the art. Any biologically-acceptable dosage form known to persons of ordinary skill in the art, and combinations thereof, are contemplated. Examples of such dosage forms include, without limitation, chewable tablets, quick dissolve tablets, effervescent tablets, reconstitutable powders, elixirs, liquids, solutions, suspensions, emulsions, tablets, multilayer tablets, bi-layer tablets, capsules, soft gelatin capsules, hard gelatin capsules, caplets, lozenges, chewable lozenges, beads, powders, gum, granules, particles, microparticles, dispersible granules, cachets, douches, suppositories, creams, topicals, inhalants, aerosol inhalants, patches, particle inhalants, implants, depot implants, ingestibles, injectables (including subcutaneous, intramuscular, intravenous, and intradermal), infusions, and combinations thereof.

[0051] Other compounds which can be included by admixture are, for example, medically inert ingredients (e.g., solid and liquid diluent), such as lactose, dextrosesaccharose, cellulose, starch or calcium phosphate for tablets or capsules, olive oil or ethyl oleate for soft capsules and water or vegetable oil for suspensions or emulsions; lubricating agents such as silica, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycols; gelling agents such as colloidal clays; thickening agents such as gum tragacanth or sodium alginate, binding agents such as starches, arabic gums, gelatin, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disintegrating agents such as starch, alginic acid, alginates or sodium starch glycolate; effervescing mixtures; dyestuff; sweeteners; wetting agents such as lecithin, polysorbates or laurylsulphates; and other therapeutically acceptable accessory ingredients, such as humectants, preservatives, buffers and antioxidants, which are known additives for such formulations.

[0052] Liquid dispersions for oral administration can be syrups, emulsions, solutions, or suspensions. The syrups can contain as a carrier, for example, saccharose or saccharose with glycerol and / or mannitol and / or sorbitol. The suspensions and the emulsions can contain a carrier, for2024-262-01example a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol.

[0053] The amount of active compound in a therapeutic composition can vary according to factors such as the disease state, age, gender, weight, patient history, risk factors, predisposition to disease, administration route, pre-existing treatment regime (e.g., possible interactions with other medications), and weight of the individual. Dosage regimens can be adjusted to provide the optimum therapeutic response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the exigencies of therapeutic situation. The molecules and compounds disclosed herein can be administered at various concentrations and through various routes depending on the intended therapeutic application and delivery methods. The dosage of the molecules and compounds disclosed herein can be adjusted based on a patient- specific factor to optimize therapeutic efficacy, therapeutic safety, or both. The patient-specific factor can include, but is not limited to, an age of a patient, body weight, body surface area, sex, overall health status, the presence an underlying medical condition, or a combination thereof. For example, a pediatric dosage can be calculated based on body weight, or body surface area to ensure appropriate dosing for a child or an adolescent. The dosage can be modified for a patient with a comorbid condition including, but not limited to, diabetes, renal impairment, hepatic dysfunction, cardiovascular disease, an immune disorder, a condition that can affect drug metabolism, a condition that can affect drug distribution, or a condition that can affect drug clearance, or a combination thereof. The dosage can be adjusted based on the severity of the bone-related condition being treated, the response of a patient to therapy, a concurrent medication, an observed adverse effect, or a combination thereof.

[0054] “Dosage unit form,’’ as used herein, refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated: each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms are2024-262-01dictated by and can be directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals. In therapeutic use for treatment of conditions in mammals (e.g., humans) for which the compounds disclosed herein, or an appropriate pharmaceutical composition thereof are effective, the compounds disclosed herein can be administered in an effective amount. The dosages as suitable for this disclosure can be a composition, a pharmaceutical composition or any other compositions described herein.

[0055] The dosage can be administered once, twice, thrice or four times a day, although more frequent dosing intervals are possible. The dosage can be administered every day, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, and / or every 7 days (once a week). The dosage can be administered daily for up to and including 30 days, preferably between 7-10 days. Or the dosage can be administered twice a day for 10 days. If the patient requires treatment for a chronic disease or condition, the dosage can be administered for as long as signs and / or symptoms persist. The patient may require “maintenance treatment” where the patient is receiving dosages every day for months, years, or the remainder of their lives. In addition, the composition can affect prophylaxis of recurring symptoms. For example, the dosage can be administered once or twice a day to prevent the onset of symptoms in patients at risk, especially for asymptomatic patients.

[0056] The compositions described herein can be administered in any of the following routes: buccal, epicutaneous, epidural, infusion, inhalation, intraarterial, intracardial, intracerebroventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, pulmonary, rectally via an enema or suppository, subcutaneous, subdermal, sublingual, transdermal, and transmucosal. The administration can be local, where the composition is administered directly, close to, in the locality, near, at, about, or in the vicinity of, the site(s) of disease, e.g., inflammation, or systemic, wherein the composition is given to the patient and passes through the body widely, thereby reaching the site(s) of disease. Local administration can be administration to the cell, tissue,2024-262-01organ, and / or organ system, which encompasses and / or is affected by the disease, and / or where the disease signs and / or symptoms are active or are likely to occur. Administration can be topical with a local effect; the composition is applied directly where its action is desired. Administration can be enteral wherein the desired effect is systemic (non-local), composition is given via the digestive tract. Administration can be parenteral, where the desired effect is systemic, composition is given by other routes than the digestive tract.

[0057] For an in vitro application, the molecules or compounds disclosed herein can demonstrate efficacy at nanomolar concentrations, with therapeutic effects observed at concentrations from about 0.3 nM to about 10 nM. The molecules or compounds disclosed herein can have effective concentrations ranging from about 0.3 nM to about 1000 nM, from about 0.3 nM to about 500 nM, from about 0.3 nM to about 100 nM, from about 1 nM to about 1000 nM, from about 10 nM to about 1000 nM, from about 100 nM to about 1000 nM, from about 100 nM to about 800 nM, from about 100 nM to about 600 nM, from about 200 nM to about 1000 nM, from about 300 nM to about 800 nM, from about 400 nM to about 700 nM, or less than, equal to, or greater than about 0.3 nM, 0.5, 1, 2. 3, 4, 5, 10, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or about 1000 nM.

[0058] For topical or local administration, the molecules or compounds disclosed herein can be incorporated into a biocompatible matrix including, but not limited to, a collagen sponge, a collagen sheet, a biodegradable scaffold, or a combination thereof. The molecules or compounds disclosed herein can be administered at concentrations from about 10 pM to about 40 pM, from about 10 pM to about 35 pM, from 10 pM to about 30 pM, from about 15 pM to about 40 pM, from about 20 pM to about 40 pM, from about 20 pM to about 35 pM, or less than, equal to, or greater than about 10 pM, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34. 36. 38. or about 40 pM. The biocompatible matrix can be soaked with a solution including the molecules or compounds disclosed herein, wherein the solution volume can be from about 20 pL to about 50 pL, from about 20 pL to about 40 pL, from about2024-262-0125 pL to about 35 pL, or less than, equal to, or greater than about 20 pL, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44. 46. 48. or about 50 pL. The biocompatible matrix can have dimensions suitable for the treatment site from about 5 cm2to about 15 cm2, from about 8 cm2to about 12 cm2, or less than equal to, or greater than 5 cm2, 6, 7, 8, 9, 10, 11, 12, 13, 14, or about 15 cm2.

[0059] For oral delivery, the molecules or compounds disclosed herein can be administered in dosages from about 1 mg / kg to about 20 mg / kg in saline solution, from about 2 mg / kg to about 15 mg / kg, from about 5 mg / kg to about 15 mg / kg, from about 8 mg / kg to about 12 mg / kg, or less than, equal to, or greater than about 1 mg / kg, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / kg.

[0060] The molecules and compounds disclosed herein can be administered alone or with one or more therapeutic agents. The molecules and compounds disclosed herein can be administered with a nucleic acidbased therapy including, but not limited to, a small interfering RNA (siRNA), a micro RNA (miRNA), an antisense oligonucleotide, a messenger RNA (mRNA), or another gene therapy vector. The nucleic acid-based therapy can target a pathway involved in bone metabolism, osteoclast differentiation, osteoblast function, an inflammatory process, or a combination thereof. The nucleic-based therapy can be administered simultaneously with, sequentially with, or separately from the molecules and compounds disclosed herein.

[0061] The molecules and compounds disclosed herein can be administered with a cell-based therapy including, but not limited to, a stem cell, a hematopoietic stem cell, a mesenchymal stem cell, a bone marrow-derived cell, a macrophage, an osteoclast precursor cell, an osteoblast precursor cell, a progenitor or differentiated cell that can contribute to bone remodeling, a progenitor or differentiated cell that can contribute to tissue repair, a progenitor or differentiated cell that can contribute to immune modulation, or a combination thereof. The cell-based therapy can be administered simultaneously with, sequentially with, or separately from the molecules and compounds disclosed herein. For example, the molecules and2024-262-01compounds disclosed herein can be incorporated into a biocompatible matrix along with a hematopoietic cell or a macrophage to provide pharmacological activation of FAK and cellular support for osteoclast differentiation and function at the treatment site.

[0062] Also contemplated herein are compositions comprising a therapeutically effective amount of one or more compounds provided herein that are useful in a method for treating any of the aforementioned musculoskeletal diseases.

[0063] The term “therapeutically effective amount” as used herein, refers to that amount of one or more compounds disclosed herein that elicits a biological or medicinal response in a tissue system, animal or human, that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated. The therapeutically effective amount can be that which may treat or alleviate the disease or symptoms of the disease at a reasonable benefit / risk ratio applicable to any medical treatment. However, it is to be understood that the total daily usage of the compounds and compositions described herein can be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically-effective dose level for any particular patient will depend upon a variety of factors, including the condition being treated and the severity of the condition; activity of the specific compound employed; the specific composition employed; the age, body weight, general health, gender and diet of the patient: the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidentally with the specific compound employed; and like factors well known to the researcher, veterinarian, medical doctor or other clinician. It is also appreciated that the therapeutically effective amount can be selected with reference to any toxicity, or other undesirable side effect, which might occur during administration of one or more of the compounds described herein.

[0064] Values expressed in a range format should be interpreted in a flexible manner to include not only the numerical values explicitly recited as2024-262-01the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range were explicitly recited. For example, a range of “about 0.1% to about 5%” or “about 0.1% to 5%” should be interpreted to include not just about 0.1% to about 5%, but also the individual values (c.g., 1%, 2%, 3%, and 4%) and the sub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the indicated range. The statement “about X to Y” has the same meaning as “about X to about Y,” unless indicated otherwise. Likewise, the statement “about X, Y, or about Z” has the same meaning as “about X, about Y, or about Z,” unless indicated otherwise.

[0065] In this document, the terms “a,” “an,” or “the” are used to include one or more than one unless the context clearly dictates otherwise. The term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Any use of section headings is intended to aid reading of the document and is not to be interpreted as limiting. Further, information that is relevant to a section heading may occur within or outside of that particular section. Furthermore, all publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference.

[0066] In the methods described herein, the steps can be carried out in any order without departing from the spirit of this disclosure, except when a temporal or operational sequence is explicitly recited. Furthermore, specified steps can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed step of doing X and a claimed step of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.

[0067] The term “about” as used herein can allow for a degree of variability in a value or range, for example, within 10%, within 5%, or within 1 % of a stated value or of a stated limit of a range.2024-262-01

[0068] The term “organic group’’ as used herein refers to any carbon-containing functional group. Examples can include an oxygen-containing group such as an alkoxy group, aralkyloxy group, a carboxyl group including a carboxylic acid, carboxylate, and a carboxylate ester; a sulfur-containing group such as an alkyl and aryl sulfide group; and other heteroatom-containing groups. Non-limiting examples of organic groups include OR, OOR, OC(O)N(R)2, CN, CF3, OCF3, R, C(O), methylenedioxy, ethylenedioxy, N(R)2, SR, SOR, SO2R, SO2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2. C(S)N(R)2, (CH2)O-2N(R)C(O)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, C(=NOR)R, and substituted or unsubstituted (Ci-Cioo)hydrocarbyl, wherein R can be hydrogen (in examples that include other carbon atoms) or a carbon-based moiety, and wherein the carbon-based moiety can be substituted or unsubstituted.

[0069] The term “substituted” as used herein in conjunction with a molecule or an organic group as defined herein refers to the state in which one or more hydrogen atoms contained therein are replaced by one or more non-hydrogen atoms. The term “functional group” or “substituent” as used herein refers to a group that can be or is substituted onto a molecule or onto an organic group. Examples of substituents or functional groups include, but are not limited to, a halogen (e.g., F, Cl, Br, and I); an oxygen atom in groups such as hydroxy groups, alkoxy groups, carboxyl groups including carboxylic acids, carboxylates, and carboxylate esters; a sulfur atom in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; a nitrogen atom in groups such as amines, hydroxyamines, nitriles, nitro groups, N-oxides, hydrazides, azides, and enamines; and other heteroatoms in various other groups. Non-limiting examples of substituents that can be bonded to a substituted carbon (or other) atom include F, Cl, Br, I, OR, OC(O)N(R)2, CN, NO, NO2, ONO2, azido, CF3, OCF3, R, O (oxo), S (thiono), C(O), S(O),2024-262-01methylenedioxy, ethylenedioxy, N(R)2. SR, SOR, SO2R, SC>2N(R)2, SO3R, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2)O-2N(R)C(0)R, (CH2)O-2N(R)N(R)2, N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R)2, N(R)SO2R, N(R)SO2N(R)2, N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R)2, N(R)C(S)N(R)2, N(COR)COR, N(OR)R, C(=NH)N(R)2, C(O)N(OR)R, and C(=NOR)R, wherein R can be hydrogen or a carbon-based moiety: for example, R can be hydrogen, (Ci-Cioo)hydrocarbyl, alkyl, acyl, cycloalkyl, aryl; or wherein two R groups bonded to a nitrogen atom or to adjacent nitrogen atoms can together with the nitrogen atom or atoms form a heterocyclyl.

[0070] The term “alkyl” as used herein refers to straight chain and branched alkyl groups and cycloalkyl groups having from 1 to 40 carbon atoms, 1 to about 20 carbon atoms, 1 to 12 carbons or, in some aspects, from 1 to 8 carbon atoms. Examples of straight chain alkyl groups include those with from 1 to 8 carbon atoms such as methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl groups. Examples of branched alkyl groups include, but are not limited to, isopropyl, iso-butyl, sec-butyl, t-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl groups. As used herein, the term “alkyl” encompasses n-alkyl, isoalkyl, and anteisoalkyl groups as well as other branched chain forms of alkyl. Representative substituted alkyl groups can be substituted one or more times with any of the groups listed herein, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups.

[0071] The term “alkenyl” as used herein refers to straight and branched chain and cyclic alkyl groups as defined herein, except that at least one double bond exists between two carbon atoms. Thus, alkenyl groups have from 2 to 40 carbon atoms, or 2 to about 20 carbon atoms, or 2 to 12 carbon atoms or, in some aspects, from 2 to 8 carbon atoms. Examples include, but are not limited to vinyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), -C(CH2CH3)=CH2, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl among others.2024-262-01

[0072] The term “alkynyl” as used herein refers to straight and branched chain alkyl groups, except that at least one triple bond exists between two carbon atoms. Thus, alkynyl groups have from 2 to 40 carbon atoms, 2 to about 20 carbon atoms, or from 2 to 12 carbons or, in some aspects, from 2 to 8 carbon atoms. Examples include, but are not limited to −C≡CH, −C≡C(CH3), −C≡C(CH2CH3), −CH2C≡CH, −CH2C≡C(CH3), and −CH2C≡C(CH2CH3) among others.

[0073] The term “acyl” as used herein refers to a group containing a carbonyl moiety wherein the group is bonded via the carbonyl carbon atom. The carbonyl carbon atom is bonded to a hydrogen forming a “formyl” group or is bonded to another carbon atom, which can be part of an alkyl, aryl, aralkyl cycloalkyl, or cycloalkylalkyl. An acyl group can include 0 to about 12, 0 to about 20, or 0 to about 40 additional carbon atoms bonded to the carbonyl group. An acyl group can include double or triple bonds within the meaning herein. An acryloyl group is an example of an acyl group. An acyl group can also include heteroatoms within the meaning herein. A nicotinoyl group (pyridyl-3 -carbonyl) is an example of an acyl group within the meaning herein. Other examples include acetyl, benzoyl, phenylacetyl, pyridylacetyl, cinnamoyl, and acryloyl groups and the like. When the group containing the carbon atom that is bonded to the carbonyl carbon atom contains a halogen, the group is termed a “haloacyl” group. An example is a trifluoroacetyl group.

[0074] The term “cycloalkyl” as used herein refers to cyclic alkyl groups such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In some aspects, the cycloalkyl group can have 3 to about 8-12 ring members, whereas in other aspects the number of ring carbon atoms range from 3 to 4, 5, 6, or 7.Cycloalkyl groups further include polycyclic cycloalkyl groups such as, but not limited to, norbornyl, adamantyl, bornyl, camphenyl, isocamphenyl, and carenyl groups, and fused rings such as, but not limited to, decalinyl, and the like. Cycloalkyl groups also include rings that are substituted with straight or branched chain alkyl groups as defined herein. Representative substituted cycloalkyl groups can be mono -substituted or substituted more than once,2024-262-01such as, but not limited to, 2,2-, 2,3-, 2,4- 2,5- or 2,6-disubstituted cyclohexyl groups or mono-, di- or tri-substituted norbornyl or cycloheptyl groups, which can be substituted with, for example, amino, hydroxy, cyano, carboxy, nitro, thio, alkoxy, and halogen groups. The term “cycloalkenyl” alone or in combination denotes a cyclic alkenyl group.

[0075] The term “aryl” as used herein refers to cyclic aromatic hydrocarbon groups that do not contain heteroatoms in the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some aspects, aryl groups contain about 6 to about 14 carbons in the ring portions of the groups. Aryl groups can be unsubstituted or substituted, as defined herein. Representative substituted aryl groups can be mono-substituted or substituted more than once, such as, but not limited to, a phenyl group substituted at any one or more of 2-, 3-, 4-, 5-, or 6-positions of the phenyl ring, or a naphthyl group substituted at any one or more of 2- to 8-positions thereof.

[0076] The term “alkoxy” as used herein refers to an oxygen atom connected to an alkyl group, including a cycloalkyl group, as are defined herein. Examples of linear alkoxy groups include but are not limited to methoxy, ethoxy, propoxy, butoxy, pentyloxy, hexyloxy, and the like.Examples of branched alkoxy include but are not limited to isopropoxy, secbutoxy, tert-butoxy, isopentyloxy, isohexyloxy, and the like. Examples of cyclic alkoxy include but are not limited to cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like. An alkoxy group can include about 1 to about 12, about 1 to about 20, or about 1 to about 40 carbon atoms bonded to the oxygen atom, and can further include double or triple bonds, and can also include heteroatoms. For example, an allyloxy group or a methoxyethoxy group is also an alkoxy group within the meaning herein, as is a methylenedioxy group in a context where two adjacent atoms of a structure are substituted therewith.

[0077] The term “amine” as used herein refers to primary, secondary, and tertiary amines having, e.g., the formula N(group)3 wherein each group2024-262-01can independently be H or non-H, such as alkyl, aryl, and the like. Amines include but are not limited to R-NH2. for example, alkylamines, arylamines, alkylarylamines; R2NH wherein each R is independently selected, such as dialkylamines, diarylamines, aralkylamines, and the like; and R3N wherein each R is independently selected, such as trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, and the like. The term “amine” also includes ammonium ions as used herein.

[0078] The term “haloalkyl” group, as used herein, includes mono-halo alkyl groups, poly-halo alkyl groups wherein all halo atoms can be the same or different, and per-halo alkyl groups, wherein all hydrogen atoms are replaced by halogen atoms, such as fluoro. Examples of haloalkyl include trifluoromethyl, 1,1 -dichloroethyl, 1,2-dichloroethyl, l,3-dibromo-3,3-difluoropropyl, perfluorobutyl, and the like.

[0079] As used herein, the term “hydrocarbyl” refers to a functional group derived from a straight chain, branched, or cyclic hydrocarbon, and can be alkyl, alkenyl, alkynyl, aryl, cycloalkyl, acyl, or any combination thereof. Hydrocarbyl groups can be shown as (Ca-Cb)hydrocarbyl, wherein a and b are integers and mean having any of a to b number of carbon atoms. For example, (Ci-C4)hydrocarbyl means the hydrocarbyl group can be methyl (Ci), ethyl (C2), propyl (C3), or butyl (C4), and (Co-Cb)hydrocarbyl means in certain aspects there is no hydrocarbyl group.

[0080] As used herein, the term “salts” and “pharmaceutically acceptable salts” refer to derivatives of the disclosed compounds wherein the parent compound is modified by making acid or base salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amines; and alkali or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, and nitric; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic.2024-262-01malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, and isethionic, and the like.

[0081] Pharmaceutically acceptable salts can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. In some instances, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two; generally, nonaqueous media like ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Lists of suitable salts are found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, the disclosure of which is hereby incorporated by reference.

[0082] The term “solvate” means a compound, or a salt thereof, that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. Where the solvent is water, the solvate is a hydrate.

[0083] EXAMPLES

[0084] The following examples are provided to illustrate the present disclosure and are not intended to limit its scope.EXAMPLE 1While searching for small molecules that would mimic a key subdomain of the N-terminal FERM domain of FAK, and therefore competitively inhibit FAK-AKT binding, compound 9 and compound 10 were identified that activated FAK at concentrations as low as 10 nM. In vitro administration of compound 9 activated FAK and promoted intestinal epithelial sheet migration in monolayer wound closure. Activation of FAK can promote osteoclastogenesis and bone resorption in vivo. For example, compound 9 increased FAK-Tyr 397 phosphorylation within intact cells, without activation of Pyk2, Pyk2 being a close paralog of FAK. Compound 9 increased FAK-Tyr 397 phosphorylation within intact cells, without2024-262-01activation of Src, Src being a non-receptor kinase within the focal adhesioncomplex.(9) (10)

[0085] The following compounds show high structural similarity tocompound 9 and suitable computationally predicted drug-like properties.(14) (15) (16)2024-262-01(17) (18) (19)

[0086] Compounds 13, 16, 21, and 22 showed scaffold similarity to compound 9 and displayed increased FAK-Tyr-379 phosphorylationcompared to a DMSO vehicle control. The rest of the compounds causedlittle change in FAK phosphorylation. Further studies performed with compounds 13, 16, 21, and 22 in Caco-2 cells showed that compounds 13,16, 21, and 22 exhibited potent dose-dependent FAK activators at nanomolar to picomolar levels. Compounds 13, 16, 21, and 22 were selected to promote Caco-2 epithelial monolayer wound closure. Compared to the DMSO vehicle control, compounds 13, 16, 21, and 22 stimulated wound closure by approximately 10-26% over baseline wound closure rates. The resultsindicated that compound 13, 16, 21, and 22 activated FAK and stimulated epithelial sheet migration.

[0087] Prompted by the enhancement of both FAK activation and invitro wound closure by compounds 13, 16, 21, and 22, synthesis of a library of FAK activators using compounds 13 and 16 were pursued as a basis for further structure-activity relationship (SAR) studies. Predictedphysiochemical properties of compounds 13, 16, 21, and 22 were calculated2024-262-01using QikProp computational software (Schrodinger, LLC). Based on scaffold similarity among the compounds, the l-(2-morpholino-5-(trifluoromethyl)phenyl)urea moiety of the FAK activator enables FAK phosphorylation. The lack of a morpholine ring in compounds 18 and 19 leads to a lack of enhanced FAK activation.

[0088] Other ring moieties were modified by rational isosteric modifications. The synthetic strategy for diversified compound preparation was based on an approach to synthesizing ureas, via treatment of a phenyl carbamate, with the corresponding cyclic or aliphatic amine derivative shown in Scheme 1.o O' tiH WrlK: / *•. X, 33< l

[0089] The compounds were tested in a biochemical assay to assess the potency to stimulate Caco-2 (by Western blotting for FAK-397) and HIEC-6 FAK activation (by a commercially available ELISA for FAK-Y-397). As seen in Table 1, on the left column, compound 13 and its analogs compound 22, 24, 16, 25,26, 27, and 28 were compared in effectiveness to phosphorylate FAK measured as fold change to FAK. In this series, compound 13 was the most potent FAK activator with a fold change value of 1.5.Table 1Table 1. Cell-based FAK-Tyr-397 activationCompound FAK- Compound FAK- Try- Tyr- 397 / FA 397 / FA K fold K fold change change2024-262-01

[0090] To enhance the solubility and FAK activation of compound 13, the hydrochloric salt compound 24 of compound 13 was synthesized.Compound 24 showed reduced FAK activation as compared to compound 13. Compound 25 was synthesized by the removal of the CF3 moiety of2024-262-01compound 13 to reduce lipophilicity. The corresponding salt compound 26 of compound 25 was synthesized. Both compound 25 and compound 26 did not induce FAK phosphorylation. Compound 27 was synthesized as the desmethoxy analog of compound 16. Compound 27 did not induce FAK phosphorylation. Compound 28 was synthesized as an analog to compound 22 bearing a hydroxy moiety to increase uptake in Caco-2 cells by glucuronidation, thus enhancing FAK phosphorylation. Compound 28 did not promote FAK phosphorylation.

[0091] Synthesizing the hydrochloric salt to produce compound 24, removing the CF3 moiety to produce compound 25, or adding hydroxyl group to produce compound 28 did not lead to increased FAK activation due to issues including, but not limited to, bioavailability, cell membrane transportation, pharmacokinetic, the like, or a combination thereof.

[0092] Compounds 13, 16, 21, and 22 increased FAK phosphorylation at low concentration and promoted the healing of intestinal epithelial wounds in vitro. Compounds 13, 16, 21, and 22 displayed drug-like properties based on both in vitro and in silico results.EXAMPLE 2

[0093] Nonsteroidal anti-inflammatory drugs (NSAIDs) comprise 7.7% of all prescriptions. 65% of NSAID users can develop upper or lower gastrointestinal (GI) ulcers. Aspirin and other NSAIDs can injure the gastroduodenal mucosa and injure the distal small bowel injury at high rates.10-40% of chronic NSAID users develop upper (GI) ulcers. 50% of chronic NSAID users develop lower GI ulcers. Medical costs of adverse GI events from NSAID use exceed $4 billion per year. NSAIDs can injure the mucosa by two different mechanisms. Proximal GI injury can reflect COX-1 inhibition. Distal small bowel injury can be caused by NSAIDs complexing to bile acids, potentiating bile acid toxicity, changing the number of enteric bacteria, changing the type of enteric bacteria, or a combination thereof. A GI erosion that is not complicated can be asymptomatic. A GI damage can cause acute bleeding, chronic anemia, perforation, stricture, obstruction, or a combination thereof. A GI ulcer can be treated by proton pump inhibitors (PPIs) or H2 blockers. However, the administration of PPIs or H2 blockers2024-262-01can reduce acid without promoting healing. Suppressing gastric can worsen NSAID small bowel enteropathy by changing the enteric microbiome.

[0094] Inflammatory bowel disease therapy can reduce inflammatory injury to stimulate mucosal healing. Mucosal healing can balance epithelial migration and cell proliferation to resurface an injured gut against continual injury, inflammation, ischemia, luminal agents, or a combination thereof.

[0095] FAK activation can be a mechanism for mucosal healing. FAK can function as a regulator of epithelial sheet migration, be a convergent target for one or more growth factors, or both. Inhibition or reduction of FAK can impair sheet migration. FAK activation can promote healing before rebleeding occurs, avoiding surgical intervention. Both total and activated FAK levels can decrease in migrating intestinal epithelial cells in vitro and in vivo at the edge of a healing ulcer, indicating that pharmacological FAK activation can facilitate the healing process. The disclosed compounds in this disclosure can target epithelial healing directly instead of neutralizing ongoing injury. The disclosed compounds in this disclosure can work in a synergistic combination with an immunosuppressive agent for treatment of inflammatory bowel disease.

[0096] In vivo studies with compound 29 demonstrated accelerated healing of a standardized ischemic intestinal ulcer. Compound 29 demonstrated accelerated healing of murine NSAID-induced distal and proximal mucosal injury, without obvious toxicity.O HNT

[0097] Compound 29 was shown to promote healing in the stomach in a chronic aspirin injury model. Compound 29 was demonstrated to act directly on FAK in an in vitro kinase assay. Compound 29 interacted and activated the 35kD kinase domain of the FAK molecule.2024-262-01

[0098] Compound 13 was tested in vitro and in vivo. In vitro studies show that compound 13 is effective at a lower concentration compared to compound 29. Compound 13 was observed to be more water soluble compared to compound 29. A therapeutic effect was observed when 10 nM. of compound 29 was administered. A therapeutic effect was observed when 0.3 nM. of compound 13 was administered.EXAMPLE 3

[0099] Follow-up structure-activity relationship (SAR) studies using commercially available analogs identified several compounds that activate FAK at nanomolar concentrations, picomolar concentrations, or both. The compounds bind the FAK catalytic domain and increase its ATP-binding affinity. Compound 30 was synthesized as water-soluble (facilitating dosing), effective at picomolar concentrations, and exhibits no cytotoxicity at lOOOOx higher than effective doses. The efficacy and non-toxicity of compound 30 has been verified in vivo, showing that it is enterally absorbed, and can be applied for patients. Compound 30 can represent a prototype therapeutic lead for an approach to heal NSAID-associated mucosal injury. Compound 30 has a short plasma half-life, which can benefit from an adjustment in QID dosing.

[0100] Compound 30, the salt of compound 31 was found to be water soluble and did not require dimethyl sulfoxide (DMSO) for solubilization. Compound 30 was observed to active FAK directly in a purified in vitro kinase assay, activate FAK in intact cells in culture, and promote epithelial monolayer wound healing in cell culture. The structures of compounds 30 and 31 are shown below:2024-262-01(30) (31)

[0101] Additional compounds 32-36 were synthesized. Compound 36 is the salt version of compound 35.-HCiEXAMPLE 4

[0102] To a stirred solution of 2-morpholino-5-trifluoromethyl aniline (0.8 g, 3.2 mmol) in dry dichloromethane (DCM) (25 mL), under N2 atmosphere at 0 °C., was added a solution of phenyl chloroformate (0.6 mL, 4.8 mmol) in dry DCM (4 mL), followed by EtsN (0.65 mL, 8.1 mmol). The crude reaction was allowed to warm up at room temperature and stirred overnight. The crude reaction was concentrated in vacuum and the product was purified by column chromatography using hexane: ethyl acetate (70: 30) to afford the pure product as a clear oil in 85 %. To a stirred solution of phenyl (2-morpholino-5-(trifluoromethyl)phenyl)carbamate (0.81 g, 2.21 mmol) in dry pyridine (5 mL) was added N, N-dimethyl-l-(pyridine-4-yl)ethane-l, 2, -diamine (0.44 g, 2.21 mmol) and the solution was heated at 80 °C. for 16 hours. The crude reaction was cooled down at room temperature and concentrated in vacuum. The product was purified by column chromatography using DCM: ethanol (95:5) to afford the pure product as an orange solid in 60% yield. l-(2-(dimethylamino)-2-(pyridin-4- 2024-262-01yl)ethyl)-3-(2-morpholino-5-(trifluoromethyl)phenyl)urea (0.438 g, 1.0mmol) was stirred in 6 mL of a 3N solution of HC1 in diethyl ether at room temperature overnight. Crude was concentrated in vacuum and lyophilized to obtain compound 30 as an orange solid in 100 % yield. Compound 37 and 38 were synthesized following the general procedure for the synthesis of compound 30.(30)

[0103] Cell culture.

[0104] Human Caco-2 cells were obtained from ATCC (Manassas, VA) and maintained in Dulbecco’s Modified Essential Eagle’s Mediumsupplemented with 10% Fetal Bovine Serum (FBS). 80-90% confluentCaco-2 cells were seeded into cell culture plates pacificated with 1% heat inactivated bovine serum albumin to prevent adhesion and avoid adhesion-associated background FAK activation. Suspended cells were then treated with DMSO (0.1%) or small molecules for 1 hour before harvesting for Western blotting.

[0105] Western blotting.

[0106] Cells were lysed and protein concentration was determined via bicinchoninic acid protein assay (Thermo Fisher, Rockford, IL). Theproteins were then separated by SDS-PAGE, transferred to nitrocellulose membranes, and incubated with antibodies to FAK-Tyr-397 (ab81298, 1:1000 dilution), or FAK (Anti-FAK, clone 4.47, 05-537, 1:1000 dilution EMD Millipore, Temecula, CA). Images were taken using a LICOR -Odyssey-Fc imaging system (LI-COR Biosciences, Lincoln, NE). Densitometry was2024-262-01conducted on exposures within the linear range, and FAK phosphorylation was calculated as the ratio of FAK-Tyr-397 intensity to FAK intensity.

[0107] Monolayer wound closure assays.

[0108] Caco-2 cells were seeded at 75-80% confluence into collagen I coated 6-well plates. Upon the cells reaching 100% confluence, wounds were made in the monolayers with non-barrier autoclaved tips. Wound images were captured using an inverted light microscope (OLYMPUS CK2, Center Valley, PA) at 0 hours and 24 hours after treatment with DMSO or small molecules. Wound areas were measured with Image J software.

[0109] Statistical analysis.

[0110] Data are depicted graphically as mean + / - standard error, and were analyzed by t-test or ANOVA as appropriate seeking 95% confidence.

[0111] A plasma pharmacokinetic study of a single dose of compound 30 was conducted using two-year-old male Fischer (CDF) rats (420-460 grams) (Chales River Laboratories, Wilmington, MA). The rats were housed under standard conditions, including a 12-hour light-dark cycle, a temperature of 22 ± 2 °C., and relative humidity of 50 ± 10%. The rats had free access to tap water and standard commercial chow. A dose of 10 mg / kg of a compound (dissolved in saline, administered in a volume of 1 mL) was given via oral gavage using Instech feed tubing (Instech, Plymouth Meeting, PA, USA). Blood samples (approximately 120 pL) were collected from the tail immediately before dosing and at 0.5, 1, 1.5, 2, 4, 6, and 10 hours postdosing using Microvette® 200 Heparin Capillary Blood Collection Tubes (Sarstedt, Numbrech, Germany). Samples were centrifuged at 2000xg for 15 minutes at 4 °C. The resulting plasma was stored at -80 °C. and analyzed by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS).

[0112] Results.

[0113] Dose-dependent half maximal effective concentration (ECso) studies in an epithelial wound closure model were performed on compounds 30, 37, and 38 using Caco-2 cells. The ECso values for the tested compounds 30, 37, and 38 were determined to be in the range of 2.36-3.01 nM., demonstrating activity in promoting epithelial wound closure. Compound 302024-262-01had a EC50 value of 3.01 nM., compound 37 had a EC50 value of 2.36 nM., and compound 38 had a EC50 value of 3.01 nM. Compound 30 was calculated to have a FAK-Tyr-397 / FAK fold change of 0.82, compound 37 was calculated to have a FAK-Tyr-397 / FAK fold change of 1.39, and compound 38 was calculated to have a FAK-Tyr-397 / FAK fold change of 1.19.

[0114] To determine whether the observed wound closure activity was due to enhanced cell migration or cell proliferation, the dose-dependent EC50 studies were repeated in the presence of 4 inM hydroxyurea, hydroxyurea being a known inhibitor of cell proliferation. The EC50 studies with hydroxyurea showed that wound closure was slowed compared to the studies conducted with hydroxyurea. The slowed wound closure indicated that cell proliferation contributed to the wound closure process. However, wound closure activity was still observed in the presence of hydroxyurea, demonstrating that the compounds promote epithelial wound closure through enhancement of epithelial cell migration.

[0115] A single dose via oral gavage at 10 mg / kg of compound 30 in saline solution, was given to two-year-old male Fisher (CDF) rates.

[0116] Two-year-old male Fischer (CDF) rats were administered a single dose of compound 30 (10 mg / kg) in saline solution via oral gavage. Blood samples were collected from the tail immediately before dosing and at 0.5, 1, 1.5, 2, 4, 6, and 10 hours post-dosing. The resulting plasma concentrations were analyzed by UPLC-MS. As seen in Figure 1, the maximum concentration (Cmax) was 967 ng / mL, after 1 hour, and the area under the curve was 3149 mg-h / E. The half-life, ti / 2, is around 3 hours. The findings indicate that compound 30 showed drug-like properties and can be a good preclinical candidate for further studies towards mucosal wound healing.EXAMPLE 5

[0117] Mice.

[0118] Wild type C57BL / 6J male mice were purchased from Jackson Laboratories (Strain #000664). The mouse colony was housed and2024-262-01maintained at Northeast Ohio Medical University (NEOMED) in a facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC). Mouse housing was maintained at 21 °C. with a 12-hour light-dark cycle. Studies presented were approved by the Institutional Animal Care and Use Committee (IACUC) at NEOMED.

[0119] Proliferation and viability.

[0120] Hematopoietic stem cells derived from flushed bone marrow of femurs and tibias of C57BL / 6J male mice at ages between 6-8 weeks. Cells were cultured in 96-well plates and seeded at 5,000 cells per well with 2% FBS while treated with L-CM for 72 hours. Proliferation was based on BrdU incorporation and quantified using CyQUANT® NF Cell Proliferation Assay Kit (Invitrogen) as per manufacturer’s instructions. Cell proliferation was assessed using fluorescent plate reader (Excitation: 485nm, Emission:530nm, Spectrum SoftMax Pro). Cell viability was assessed using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) colorimetric assy. Cells were plated and assessed using the CellTiter 96® Aqueous One Solution Assay protocol (Promega). Plates were incubated with CellTiter 96® and read at 490nm using a 96-well plate reader (Spectrum SoftMax Pro).

[0121] Isolation of Primary osteoclasts and Differentiation.

[0122] Murine bone marrow cells were flushed and collected from femurs and tibias of C57BL / 6J male mice at ages between 6-8 weeks. Nonadherent cells were collected and seeded on 96-well plates or bone slices at 200,000 cells / well while exposed to 50 ng / mL of M-CSF. After 3 days, macrophages were treated with various doses of L-CM based on the experiment with 50 ng / mL of RANK-L. Treatment and RANK-L doses were replenished every other day until the osteoclasts matured.

[0123] Tartrate-Resistant Acid Phosphatase (TRAP) Activity and Staining.

[0124] Osteoclast cultures were assessed for TRAP staining and activity. Mature osteoclasts were fixed with 10% formalin for 10 minutes2024-262-01and washed twice with phosphate-buffered saline (PBS). For TRAP activity, methanol: acetone (1:1) was added to each well followed by TRAP buffer (52 mm of Na-tartrate in 0.1 M Na-acetate buffer, pH 5.2) containing 0.1 mg / mL of p-nitrophenyl phosphate. After 30 minutes of incubation at 37° C., reaction was stopped with IN NaOH and optical density was assessed at 405nm using 96-well plate reader (Spectrum SoftMax Pro). For TRAP staining, wells were washed twice with PBS, followed by incubation in TRAP buffer with 1.5 mm naphthol AS-MX phosphate and 0.5 mm Fast Red Violet LB Salt for 30-60 minutes at 37° C. The TRAP stain was then washed twice with PBS and cells were left submerged in 60% glycerol PBS until imaged. TRAP-positive osteoclasts (>3 nuclei) were imaged using a Nikon Ti Eclipse inverted microscope (Nikon Instruments Inc., Melville, NY, USA). Osteoclast number, size, and differential count were calculated using the NIS-Elements software (Nikon Instruments Inc.).

[0125] Pit Resorption Assay.

[0126] Osteoclasts were seeded on the surface of bone cortical slices (bone slices, BioVendor) and differentiated for 12-14 days. Following fixation, bone slices were TRAP stained, imaged, and counted. Following TRAP stain, bone slices were sonicated for 1 hour and brushed to remove osteoclasts. Resorption side of bone slices was exposed to 1% toluidine blue to stain for pit resorption followed by 5x H2O washes. Pit resorption area was imaged using NIS-Elements software (Nikon Instruments Inc.) and quantified using ImageJ.

[0127] Clavaria Bone Resorption Assay.

[0128] C57BL / 6J mice were used to assess in vivo bone resorption. Fivc-wcck-old animals were anesthetized with isofluranc and their calvarias exposed. Animals were treated with collagen sponges (10cm2, Medline) soaked in 30uL of PBS (Controls), Medium + RANK-L (50pg / mL), or different concentrations of FAK activator (Compound 30) (n=4-6 per group). After 7 days, animals were euthanized with CO2 immersion and their calvariae fixed in 10% formalin overnight. Following washing and trimming, calvarias were stained for TRAP as indicated above. TRAP+ osteoclasts and resorptive area was quantified using Osteomeasure software and imaged2024-262-01using Nikon SMZ 800 stereomicroscope (Nikon Instruments Inc.). Three-dimensional reconstruction was performed through micro-CT scan using SkyScan CTvox software (Skyscan 1172, Microphotonics, PA).

[0129] Data and Statistical Analysis.

[0130] Data were analyzed using GraphPad Prism 9 software (GraphPad). All individual experiments were repeated at a minimum of n=3 per experiment with at least 3-6 replicates per experiment. One-way ANOVA analysis followed by Dunnett’s post-hoc test was performed when comparing multiple groups. An unpaired t-test was performed to compare two groups. Differences were considered statistically significant if the p value was less than 0.05.

[0131] Results.

[0132] Hematopoietic stem cells were harvested from 6-8 week old C57 / B16 male mice, cultured with macrophage colony-stimulating factor- 1 (MSCF-1) (30 ng / ml) and FAK activator compound 30 at doses ranging from 100-1000 nM. Cultures were terminated 48 hours later for the assessment of cell viability and proliferation. Compound 30 treatment did not substantially affect hematopoietic stem cell viability, as shown in FIG. 2A. Compound 30 treatment did not substantially affect hematopoietic stem cell proliferation, as shown in FIG. 2B.

[0133] Bone marrow-derived hematopoietic stem cells were treated with MCSF to induce macrophage differentiation, then treated with both MCSF and RANKL to induce osteoclast differentiation. Cells were left untreated (control) or treated with different concentrations of compound 30. After 6-7 days in culture, osteoclasts were stained for TRAP (marker for osteoclast differentiation) and assessed for TRAP activity and osteoclasts were counted. The stained osteoclasts is shown in FIG. 3.

[0134] The administration of compound 30 did not affect TRAP activity significantly, as shown in FIG. 4A. The was an increase in the total osteoclast count when compound 30 was administered to the bone marrow-derived hematopoietic stem cells. There was a significant increase in the2024-262-01total osteoclast count when 500 nM. of compound 30 was administer to the bone marrow-derived hematopoietic stem cells.

[0135] FIG. 4B shows that when 500 nM. of compound 30 was administered, there was less TRAP activity. FIG. 4C shows that treatment with compound 30 demonstrated less TRAP activity per osteoclast.

[0136] Differential counts were performed, counting osteoclasts with 3-10 nuclei, 11-20 nuclei, or more than 20 nuclei per cell. FIG. 5A-5C show that treatment with compound 30 stimulates osteoclast differentiation.

[0137] For in vitro analysis, osteoclasts seeded on bone slices were differentiated while treated with various doses of compound 30 when compared to untreated control. FIG. 6A shows osteoclasts stained with TRAP. Following TRAP staining remove, resorption pits were stained using toluidine blue. FIG. 6B shows osteoclasts stained with toluidine blue. FIG.7 A demonstrates that treatment with compound 30 did not have a significant effect on total osteoclast counts in cultures of osteoclasts on bone chips in the presence of MCSF and RANKL. However, FIG. 7B shows that the function of the osteoclasts was enhanced in a dose-dependent manner when compound 30 was administered.

[0138] Male mice around 6 weeks of age, were subjected to in vivo bone resorption assay using the calvaria (skull) model. A collagen sheet was used a carrier for RANKL and compound 30. Mice received collagen sheets soaked with either (lOOng / ml) RANKL (to induce osteoclastogenesis) with PBS (control) or collagen sheets soaked with RANKL and 10-40 pM of compound 30 (n=6 per condition). After one week, the mice were sacrificed and the calvariae were harvested, cleaned and stained for TRAP. The total number of osteoclasts was counted and their size recorded. FIG. 8 shows calvariae that were cleaned, fixed, and stained for TRAP. Each round cell in FIG. 8 represents an osteoclast.

[0139] FIG. 9 shows the quantification of osteoclast number and resorption area. FIG. 9 shows a decreasing number of osteoclasts and decreasing osteoclast size in response to treatment with compound 30.2024-262-01

[0140] The results suggest that the FAK activator, compound 30, stimulates osteoclast differentiation and function as well bone resorption in vivo. There is evidence that FAK activation influences osteoblast differentiation and bone regeneration.

[0141] All publications, patents and patent applications are incorporated herein by reference. While in the foregoing specification this disclosure has been described in relation to certain preferred aspects thereof, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the disclosure is susceptible to additional aspects and that certain of the details described herein may be varied considerably without departing from the basic principles of the disclosure.EXEMPLARY ASPECTS

[0142] The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the embodiments of the present disclosure. Thus, it should be understood that although the present disclosure has been specifically disclosed by specific embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those of ordinary skill in the art, and that such modifications and variations are considered to be within the scope of embodiments of the present disclosure.

[0143] The following exemplary aspects are provided, the numbering of which is not to be construed as designating levels of importance.

[0144] Aspect 1 provides a molecule used to treat a musculoskeletal disease comprising a compound of Formula 1 or a salt thereof,2024-262-01R1(1),whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -H, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.

[0145] Aspect 2 provides the molecule of Aspect 1, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl. (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0146] Aspect 3 provides the molecule of Aspect 1, wherein the compound is represented by Formula 2:2024-262-01RR1(2), whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -H, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof.

[0147] Aspect 4 provides the molecule of Aspect 3, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0148] Aspect 5 provides the molecule of Aspect 1, wherein the compound is represented by Formula 3:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH₂CH₂OCH₂, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

[0149] Aspect 6 provides the molecule of Aspect 1, wherein the compound is represented by Formula 4:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CFhCFfcj-OCFk, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

[0150] Aspect 7 provides the molecule of Aspect 1, wherein the compound is represented by Formula 5:

[0151] Aspect 8 provides the molecule of Aspect 1, wherein the compound is represented by Formula 6:2024-262-01(6), wherein n is any positive integer.

[0152] Aspect 9 provides the molecule of Aspect 1, wherein the compound is represented by Formula 7:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF₃, -CH3, -CH2CH3.-OCH3. -OCH2CH3, and combinations thereof.

[0153] Aspect 10 provides the molecule of Aspect 1, wherein the compound is represented by Formula 8:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH₂CH₂OCH₂, -OCH2CH3, and combinations thereof.

[0154] Aspect 11 provides a method to treat a musculoskeletal disease comprising administering to a subject in need thereof an effective amount of a compound of Formula 1 or a salt thereof,whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -II, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.2024-262-01

[0155] Aspect 12 provides the method of Aspect 11, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0156] Aspect 13 provides the method of Aspect 11, wherein the compound is represented by Formula 2:R1R1(2), whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -H, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof.

[0157] Aspect 14 provides the method of Aspect 13, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

[0158] Aspect 15 provides the method of Aspect 11, wherein the compound is represented by Formula 3:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH₂CH₂OCH₂, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

[0159] Aspect 16 provides the method of Aspect 11, wherein the compound is represented by Formula 4:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH₂CH₂OCH₂, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

[0160] Aspect 17 provides the method of Aspect 11, wherein the compound is represented by Formula 5:2024-262-01

[0161] Aspect 18 provides the method of Aspect 11, wherein the compound is represented by Formula 6:wherein n is any positive integer.

[0162] Aspect 19 provides the method of Aspect 11, wherein the compound is represented by Formula 7:2024-262-01wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3.-OCH3. -OCH2CH3, and combinations thereof.

[0163] Aspect 20 provides the method of Aspect 11, wherein the compound is represented by Formula 8:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3.-OCH3. -OCH2CH3, and combinations thereof.2024-262-01

Claims

CLAIMSWhat is claimed is:

1. A molecule used to treat a musculoskeletal disease comprising a compound of Formula 1 or a salt thereof,whereinX1is -C-, -N-, or -O-;X2is -N- or -C-: andR1and R2are independently selected from the group consisting of -H, -OH, and substituted or unsubstituted (Ci-C2o)hydrocarbyl, and combinations thereof.

2. The molecule of claim 1, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

3. The molecule of claim 1, wherein the compound is represented by Formula 2:2024-262-01R1(2), whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -II, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof.

4. The molecule of claim 3, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl. (Ci-C2o)aryl. (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

5. The molecule of claim 1, wherein the compound is represented by Formula 3:2024-262-01whereinat each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

6. The molecule of claim 1, wherein the compound is represented by Formula 4:2024-262-011wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

7. The molecule of claim 1, wherein the compound is represented by Formula 5:2024-262-018. The molecule of claim 1, wherein the compound is represented by Formula 6:wherein n is any positive integer.2024-262-019. The molecule of claim 1, wherein the compound is represented by Formula 7:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

10. The molecule of claim 1, wherein the compound is represented by Formula 8:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.2024-262-0111. A method to treat a musculoskeletal disease comprising administering to a subject in need thereof an effective amount of a compound of Formula 1 or a salt thereof,whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -H, -OH, and substituted or unsubstituted (Ci-Cbojhydrocarbyl, and combinations thereof.

12. The method of claim 11, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ca-Czojalkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl, (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

13. The method of claim 11, wherein the compound is represented by Formula 2:2024-262-01R1(2), whereinX1is -C-, -N-, or -O-;X2is -N- or -C-; andR1and R2are independently selected from the group consisting of -II, -F, -OH, substituted or unsubstituted (C1-C20) hydrocarbyl, and combinations thereof.

14. The method of claim 13, wherein the (Ci-C2o)hydrocarbyl is selected from the group consisting of (Ci-C2o)alkyl, (Ci-C2o)alkenyl, (Ci-C2o)alkynyl, (Ci-C2o)acyl, (Ci-C2o)cycloalkyl. (Ci-C2o)aryl, (Ci-C2o)alkoxy, (Ci-C2o)haloalkyl, and combinations thereof.

15. The method of claim 11, wherein the compound is represented by Formula 3:2024-262-01whereinat each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -C-, -N-, -O-, and combinations thereof.

16. The method of claim 11, wherein the compound is represented by Formula 4:2024-262-01whereinat each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof; andY1, Y2, and Y3are independently selected from the group consisting of -N-, -C-, -O-, and combinations thereof.

17. The method of claim 11, wherein the compound is represented by Formula 5:2024-262-0118. The method of claim 11, wherein the compound is represented by Formula 6:wherein n is any positive integer.2024-262-0119. The method of claim 11, wherein the compound is represented by Formula 7:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.

20. The method of claim 11, wherein the compound is represented by Formula 8:wherein at each occurrence, R1is independently selected from the group consisting of -H, -F, -CF3, -CH3, -CH2CH3,-OCH3, -OCH2CH3, and combinations thereof.2024-262-01

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