Compositions and methods for treating tendon and bone injuries

Abstract

In one aspect, the invention relates to synthetic molecules, compositions and pharmaceutical compositions, which are useful for delivering a bioactive moiety to a site of injun'. Also disclosed are methods of treating a tendon injury, methods of treating a fracture, methods of treating a bone stress injury, methods of enhancing incorporation of allograft tissues, methods of promoting repair or regeneration of enthesis tissue during or after an orthopedic surgical procedure, and method of treating osteoporosis, osteoarthritis and / or cartilage defects using the disclosed molecules, compositions and pharmaceutical compositions.

Description

ATTORNEY DOCKET NO.37759.0682P1COMPOSITIONS AND METHODS FOR TREATING TENDON AND BONE INJURIESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U. S. Provisional Patent Application No. 63 / 755,772, filed on February 7. 2025, which is incorporated by reference herein in its entirety.SEQUENCE LSITING

[0002] The Sequence Listing submitted as a text file named “37759.0682P1’" created on February 5, 2026, and having a size of 22,792 bytes is hereby incorporated by reference.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0003] This invention was made with government support under IK2BX005199 awarded by the Department of Veterans Affairs. The government has certain rights in the invention.BACKGROUND

[0004] There are 32 million musculoskeletal injuries in the United States annually of which 45% involve tendons or ligaments such as rotator cuff tendon tears and anterior cruciate ligament (ACL) tears. These two common injuries that are often treated surgically and constitute a significant healthcare burden in the United States. Approximately 250,000 RCT repairs are performed each year in the US alone and it has been estimated that successful rotator cuff repairs in the US result in a lifetime societal savings of $3.44 billion. Unfortunately, on average 12% to 40% of rotator cuff repairs will fail to heal, and failure of up to 94% of repairs has been reported when rotator cuff tears are larger than 2cm in size or affect patients greater than 60 years of age, which is a noteworthy challenge since both of these factors are commonly encountered in clinical practice. A critical feature related to this high rate of observed tendon repair failure is that, currently, tendon-to-bone repairs heal via the formation of disorganized fibrotic tissue that fails to restore the native tissue architecture, and this abnormal reparative tissue is compositionally and mechanically inferior to the native tendon and enthesis tissue.

[0005] In general, the biology of tissue regeneration involves cells, extracellular matrix (ECM) and growth factors. Cells are in relative abundance given the close proximity of decorticated bone, periosteum, bursal adipose and often synovial tissues at the time of aATTORNEY DOCKET NO.37759.0682P1tendon-bone repair and the available pool of local mesenchymal progenitor cells are certainly capable of proliferating and producing ECM when appropriately exposed to various trophic factors. While certain growth factors such as bone morphogenetic proteins (BMPs) or transforming growth factor-beta (TGF(3) have been reported to improve both the biomechanical strength and tissue organization of the repaired enthesis. their clinical applicability continues to be severely limited due to a lack of effective delivery strategies.

[0006] Delivering growth factors to the site of tendon repair is an attractive strategy to incite the ingress of local progenitor cells and trigger tissue regeneration. Current clinical approaches to growth factor delivery are limited to supraphysiologic doses administered at the gross tissue level at one point in time (i.e. intra-operative delivery). Unfortunately, there is no current methodology to i) effectively deliver physiologically -relevant levels of growth factors at the cellular level, ii) target delivery to the site of surgical repair, and iii) deliver growth factors over multiple time points.

[0007] What is needed are molecules, compositions and pharmaceutical compositions and methods for targeting a molecule to a site of injury or site of surgical repair. Also needed are methods of treating a tendon injury, methods of treating a fracture, methods of treating an osteotomy and methods of treating a bone stress injury.BRIEF SUMMARY

[0008] Described herein are molecules, compositions and pharmaceutical compositions as well as methods for targeting a molecule to a site of injury. Further described herein are molecules, compositions and pharmaceutical compositions for treating a tendon injury, a fracture, and / or a bone stress injury. Also described herein are molecules, compositions and pharmaceutical compositions for enhancing incorporation of allograft tissues and for promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure. Also described herein are molecules, compositions and pharmaceutical compositions for treating osteoporosis, osteoarthritis and / or cartilage defects and reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site.

[0009] Disclosed herein are molecules comprising i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0010] Disclosed herein are compositions comprising a molecule, wherein the molecule comprises a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety;ATTORNEY DOCKET NO.37759.0682P1wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0011] Disclosed herein are pharmaceutical compositions comprising a molecule, wherein the molecule comprises i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0012] Disclosed herein are methods of targeting a molecule to a site of injury comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0013] Disclosed herein are methods of treating a tendon injury, a fracture, and / or a bone stress injury and / or promoting soft tissue to bone healing comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety'; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0014] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety7; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0015] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety7.

[0016] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0017] Disclosed herein are methods of reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site comprising administering a molecule, wherein the molecule comprises i) a bioactive moiety; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein theATTORNEY DOCKET NO.37759.0682P1cleavable linker links the bioactive moiety to the BP moiety.

[0018] Disclosed herein are molecules comprising i) a variant of BMP-2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0019] Disclosed herein are compositions comprising a molecule, wherein the molecule comprises a variant of BMP-2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0020] Disclosed herein are pharmaceutical compositions comprising a molecule, wherein the molecule comprises i) a variant of BMP -2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0021] Disclosed herein are methods of targeting a molecule to a site of injury comprising administering a molecule, wherein the molecule comprises i) a variant of BMP-2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0022] Disclosed herein are methods of treating a tendon injury, a fracture, and / or a bone stress injury and / or promoting soft tissue to bone healing comprising administering a molecule, wherein the molecule comprises i) a variant of BMP -2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP -2 to the BP moiety.

[0023] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a variant of BMP -2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0024] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a variant of BMP -2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0025] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects comprising administering a molecule, wherein the molecule comprises i) a variant of BMP-2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein theATTORNEY DOCKET NO.37759.0682P1BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP -2 to the BP moiety.

[0026] Disclosed herein are methods of reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site comprising administering a molecule, wherein the molecule comprises i) a variant of BMP -2; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0027] Disclosed herein are molecules comprising i) a piezol agonist, an IL-1 (3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1(3 antagonist, IRAK.4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0028] Disclosed herein are compositions comprising a molecule, wherein the molecule comprises a piezol agonist, an IL- 1 (3 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety7has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1(3 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0029] Disclosed herein are pharmaceutical compositions comprising a molecule, wherein the molecule comprises i) a piezol agonist, an IL- 1(3 antagonist, an IRAK.4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1(3 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0030] Disclosed herein are methods of targeting a molecule to a site of injury comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL-1(3 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1 (3 antagonist, IRAK.4 inhibitor, caspase-1ATTORNEY DOCKET NO.37759.0682P1inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0031] Disclosed herein are methods of treating a tendon injury, a fracture, and / or a bone stress injury and / or promoting soft tissue to bone healing comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL-ip antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and hi) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0032] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL- 1 P antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and hi) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1 P antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety7.

[0033] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL-1 P antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL-1 P antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0034] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL- ip antagonist, an IRAK.4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety; wherein the BP moiety' has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1 P antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.ATTORNEY DOCKET NO.37759.0682P1

[0035] Disclosed herein are methods of reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site comprising administering a molecule, wherein the molecule comprises i) a piezol agonist, an IL- 1 [3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor; ii) a cleavable linker; and iii) a bisphosphonate (BP) moiety'; wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the piezol agonist, IL- 1|3 antagonist, IRAK.4 inhibitor, caspase- 1 inhibitor. NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several aspects of the disclosed method and compositions and together with the description, serve to explain the principles of the disclosed method and compositions.

[0037] FIG. 1 shows H& E stained images shown under bright field in the top row and under polarized light in the bottom row. The images show critical enthesis between tendon and bone is not regenerated during the healing process.

[0038] FIG. 2 is a schematic of BP-BMP2 drug delivery’ strategy.

[0039] FIGs. 3A-3D show surgery on mouse (A) Transosseous and transtendinous stitch placement, (B) sharp transection of Achilles tendon at bone attachment, (C) posteriorsuperior calcaneus burr for decortication, and (D) repair of transected tendon to decorticated calcaneus.

[0040] FIGs. 4A-4C show results from AF647 experimental cohort. Mean fluorescence on-target signal (A) and off-target signal (B) over time among systemic, local and sham treatment groups (n = 7 per group) yvho received AF647-ZOL. Error bars represent standard deviation from the mean value. Both ** and * denote a significant difference between Sham and Local groups (p < 0.01, p < 0.05), respectively. Both ## and # denote a significant difference between Sham and Systemic groups (p < 0.01, p < 0.05), respectively. (C) Representative image of fluorescence signal on POD 10 among animals treated yvith AF647-ZOL

[0041] FIGs. 5A-5C show results from Cat-K 680 experimental cohort. Mean fluorescence on-target signal (A) and off-target signal (B) over time among repair (systemic) and sham treatment groups (n = 3 per group) yvho received Cat K 680 FAST. Error barsATTORNEY DOCKET NO.37759.0682P1represent standard deviation from the mean value. ** and * denote a significant difference between Sham and Repair groups (p < 0.01, p < 0.05). respectively. (C) Representative image of fluorescence signal on POD 4 among sham and Achilles tendon repair animals treated with Cat K 680 FAST.

[0042] FIGs. 6A-6C show results from OFS-3 experimental cohort. OFS-3 is a FRET-quenched molecular probe providing a similar signal activation mechanism to Cat-K 680 but including a bone-targeting BP moiety. Mean fluorescence on-target signal (A) and off-target signal (B) over time among systemic, local and sham treatment groups (n = 7 per group) who received OFS-3. Error bars represent standard deviation from the mean value. Both ** and * denote a significant difference between Sham and Local groups (p < 0.01, p < 0.05), respectively. Both ## and # denote a significant difference between Sham and Systemic groups (p < 0.01, p < 0.05), respectively. (C) Representative in vivo fluorescent imaging at postoperative day 10 for local, systemic, and sham treatment groups.

[0043] FIGs. 7A-7B show histological analysis of OFS-3 treatment and control groups. (A) H+E results: Mouse hindfoot sagittal sections taken 4 and 6 weeks after Achilles tendon-to-bone repair followed by saline (AR) and Achilles tendon-to-bone repair followed by application of OFS-3 (AR + OFS-3), stained with H& E. (B) IHC results: Mouse hindfoot sagittal sections taken 4and 6 weeks after Achilles tendon-to-bone repair followed by saline (AR) or OFS-3 (AR + OFS-3) treatment, stained with immunohistochemistry for Ctsk. Anti-Ctsk antibodies are stained brown while blue stain reflects residual hematoxylin.

[0044] FIG. 8A-8B show plasmid maps for bacterial expression vectors. A) pETl 1 a-pyltRNA containing mutated BMP2 sequence, where the codon (AAA) coding for Lys3 in wild-type sequence was replaced by TAG (amber-stop-codon), and TAG stop codon was replaced by a TAA stop codon; B) for pRSFduet-pyrtRNAsynth, which encodes the corresponding aminoacyl-tRNA synthetase.

[0045] FIG. 9 shows the structure of an OTS linker. The peptide linker shown is Ctsk-sensitive linker GHPGGPQG (SEQ ID NO: 1).

[0046] FIG. 10 shows time-course experiment for the CuAAC-mediated bioconjugation of the OTS linker and BMP2K3Plk. Each lane represents a time-point (minutes). The starting material monomer has an approximate mass of 13 kDa and the starting material dimer has an approximate mass of 26 kDa. The target conjugate has an expected mass of approximately 28 kDa (26,000 + 2 * 1200 = 28.4 kDa). Protein fragmentation (9 kDa) and multimer formation (40 kDa) during CuAAC-mediated bioconjugations using BMP2-K3Plk is shown

[0047] FIG. 11A-11C shows characterization of OTS-BMP2 conjugate. (A) Gel analysisATTORNEY DOCKET NO.37759.0682P1of product mixture with three different loading volumes (15, 7.5 and 3.75 pL). The % fragmented protein of total protein is given at the bottom of each lane. (B) Mass spectrum (ESI)+ of monomeric OTS-BMP2 (TCEP treated). A m / z ratio was observed of 12- (1188.40 m / z, calcd [M+12H]12+: 1188.46 m / z), 10- (1425.7784 m / z, calcd [M+10H] 10+: 1425.95 m / z), 9- (1584.0864 m / z, calcd [M+9H]9+: 1584.28 m / z) charged ions for monomeric OTS-BMP2. Undefined peaks in the 500 - 1000 m / z range likely corresponding to fragmented protein was also observed. (C) Deconvoluted mass spectrum of monomeric OTS-BMP2 (TCEP treated) observe dominant mass 14249.46 Da, expected 14249.47 Da.

[0048] FIG. 12 shows results from a BRE expression assay. FIG. 12 shows that bisphosphonate bound variant BMP2 does not activate BMP pathways among ATDC5cells when BP-BMP2 is not exposed to / digested by cathepsin K.

[0049] FIG. 13 shows pCT images of 5 different rat shoulders after rotator cuff injury and repair at 3 different time points (1 week, 3 weeks, and 5 weeks) after surgery.

[0050] FIG. 14 shows load to failure biomechanical testing of the repaired supraspinatus tendon at 6 weeks post-injury and 6 weeks post-treatment. BP-BMP2 (OTS-BMP2) at different doses (0.5pg, lug, 5pg, and lOpg) administered on post-operative day 0.

[0051] FIG. 15 shows stiffness biomechanical testing of the repaired supraspinatus tendon at 6 weeks post-injury' and 6 weeks post-treatment with subcutaneous BP-BMP2 (OTS-BMP2) at 3 different doses (0.5pg, Ipg, 5pg) administered on post-operative day 0.

[0052] FIG. 16 shows hematoxylin and eosin staining of histologic specimens 6 weeks after rat rotator cuff tendon tear and repair. Rats were treated with either saline or 5ug BP-BMP2 (OTS-BMP2) administered subcutaneously on post-operative day 0 (POD 0).

[0053] FIGs. 17A-D show operative limb pressure index (relative luminance divided by¬ paw surface area for each animal), which is a surrogate for weight bearing, as measured using BlackBox R4 device (Blackbox Biotech Inc.). (FIG. 17A) 2 weeks after surgery, (FIG. 17B) 3 weeks after surgery, (FIG. 17C) four weeks after surgery, and (FIG. 17D) six weeks after surgery, 5x dose = 5pg; lOx dose = lOpg; POW = post-operative week; POD = postoperative day.

[0054] FIGs. 18A-B show OFS-3 administration among DMM treated C57BL / 6 mice vs. sham over time. FIG. 18A shows mean fluorescence change from day before surgery of both DMM and sham treatment. FIG. 18B shows mean fluorescence difference (DMM-Sham). Post-operative day (x-axis) in these figures reflects the days after the injection procedure, thus would be better characterized as post-injection day. The animal underwent DMM surgery' 10 weeks prior to administration of OFS-3. Sham group consisted of n=5 animals,ATTORNEY DOCKET NO.37759.0682P1and the OFS-3 group consisted of n=8 animals. Y-axis is on a log scale.DETAILED DESCRIPTION

[0055] The disclosed method and compositions may be understood more readily by reference to the following detailed description of particular aspects and the Examples included therein and to the Figures and their previous and following description.

[0056] It is to be understood that the disclosed method and compositions are not limited to specific synthetic methods, specific analytical techniques, or to particular reagents unless otherwise specified, and, as such, may vary. It is also to be understood that the terminology7used herein is for the purpose of describing particular aspects only and is not intended to be limiting.A. Definitions

[0057] The terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting.

[0058] As used in the specification and the appended claims, the singular forms “a,” "an" and “the’’ can include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a compound” includes mixtures of compounds, reference to “a pharmaceutical carrier” includes mixtures of two or more such carriers, and the like.

[0059] The w ord “or” as used herein means any one member of a particular list and also includes any combination of members of that list.

[0060] Ranges may be expressed herein as from “about” one particular value, and / or to “about” another particular value. The term "about" is used herein to mean approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. In general, the term “about” is used herein to modify a numerical value above and below7the stated value by a variance of 20%. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

[0061] As used herein, the term “amino acid sequence” refers to a list of abbreviations, letters, characters or words representing amino acid residues. The amino acid abbreviations used herein are conventional one letter codes for the amino acids and are expressed asATTORNEY DOCKET NO.37759.0682P1follows: A, alanine; C, cysteine; D aspartic acid; E, glutamic acid; F, phenylalanine; G, glycine; H histidine; I isoleucine; K. lysine; L, leucine; M, methionine; N, asparagine; P. proline; Q, glutamine; R, arginine; S, serine; T, threonine; V, valine; W, tryptophan; and Y, tyrosine. All alpha amino acids except glycine can exist in either of two enantiomers, called L-amino acids or D-amino acids, which are mirror images of each other.

[0062] As used herein, the term “amino acid” refers to an organic molecule with a basic amino group (-NH2), an acidic carboxyl group (-COOH), a hydrogen and a variable ‘R’ group bound to an sp3 hybridized central carbon atom. The N-terminus of a peptide has a free amino group (-NH2). The C-terminus of a peptide has a free carboxyl group ( COOH).

[0063] As used herein, the terms “D-amino acid” or “amino acid in the D-form” refer to amino acids where the stereogenic carbon alpha to the amino group has the D-configuration. D amino acid is the enantiomer of an amino acid that is capable of rotating plane polarized light clockwise (right-hand side). D-amino acids are designated with the prime (’) symbol.

[0064] As used herein, the terms “L-amino acid” or “amino acid in the L-form” refer to amino acids where the stereogenic carbon alpha to the amino group has the L-configuration. L amino acid is the enantiomer of an amino acid that is capable of rotating plane polarized light anticlockwise (left-hand side). With the exception of achiral glycine, natural amino acids have the L configuration.

[0065] “Peptide” as used herein refers to any peptide, oligopeptide, polypeptide, gene product, expression product, or protein. A peptide is comprised of consecutive amino acids. The term “peptide” encompasses naturally occurring or synthetic molecules. A residue of a peptide is an amino acid. As used herein if an amino acid is said to be in the 5thposition, it is the 5thamino acid from the N-terminus of the peptide.

[0066] By “fragment” is meant a portion of a polypeptide or nucleic acid molecule, such as, but not limited to, a truncation mutant. This portion contains, preferably, at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% of the entire length of the reference nucleic acid molecule or polypeptide. For example, a fragment of a polypeptide may contain about 5. about 10, about 15, about 20, about 25 or about 30 or more amino acids. In another example, a fragment of a polypeptide may contain about 1000, about 1500, about 2000, about 2500 or about 3000 or more amino acids.

[0067] As used herein, the term “epitope” refers to a localized region on the surface of an antigen capable of eliciting an immune response.

[0068] As used herein, “sample” is meant to mean an animal; a tissue or organ from an animal; a cell (either within a subject, taken directly from a subject, or a cell maintained inATTORNEY DOCKET NO.37759.0682P1culture or from a cultured cell line); a cell lysate (or lysate fraction) or cell extract; or a solution containing one or more molecules derived from a cell or cellular material (e.g. a polypeptide or nucleic acid), which is assayed as described herein. A sample may also be any body fluid or excretion (for example, but not limited to, blood, urine, stool, saliva, tears, bile) that contains cells or cell components.

[0069] As used herein, “subject” refers to the target of administration, e.g. an animal. Thus, the subject of the disclosed methods can be a vertebrate, such as a mammal. For example, the subject can be a human. The term does not denote a particular age or sex.Subject can be used interchangeably with “individual” or “patient”.

[0070] Substantial changes in function or immunological identity are made by selecting substitutions that are less conservative than those in Table 1, i.e., selecting residues that differ more significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain. The substitutions which in general are expected to produce the greatest changes in the protein properties are those in which: (a) the hydrophilic residue, e.g. seryl or threonyl, is substituted for (or by) a hydrophobic residue, e.g., leucyl, isoleucyl, phenylalanyl, valyl or alanyl;Tryptophan, Tyrosinyl (b) a cysteine or proline is substituted for (or by) any other residue; (c) a residue having an electropositive side chain, e g., lysyl, arginyl, or hystidyl, is substituted for (or by) an electronegative residue, e.g. glutamyl or aspartyl; or (d) a residue having a bulky side chain, e g., phenylalanine, is substituted for (or by) one not having a side chain, e.g., glycine, in this case, or (e) by increasing the number of sites for sulfation and / or glycosylation.Table 1 Amino Acid SubstitutionsNon-limiting ExemplaryOriginal ResidueConservative SubstitutionsAla SerArg Gly; Gin; LysAsn Gin; HisAsp GluCys SerGin Asn; LysGlu AspATTORNEY DOCKET NO.37759.0682P1Non-limiting ExemplaryOriginal ResidueConservative SubstitutionsGly AlaHis Asn; GinIle Leu; ValLeu Ile; ValLys Arg; GinMet Leu; IlePhe Met; Leu; TyrSer ThrThr SerTrp TyrTyr Trp; PheVal Ile; LeuTable 2: Amino Acid AbbreviationsAmino Acid AbbreviationsAlanine Ala (A)Allosoleucine AIleArginine Arg (R)Asparagine Asn (N)Aspartic Acid Asp (D)Cysteine Cys (C)Glutamic Acid Glu (E)Glutamine Gin (Q)Glycine Gly (G)Histidine His (H)Isolelucine Ile (I)Leucine Leu (L)Lysine Lys (K)Phenylalanine Phe (F)Praline Pro (P)Pyroglutamic Acid PGlu (U)ATTORNEY DOCKET NO.37759.0682P1Amino Acid AbbreviationsSerine Ser (S)Threonine Thr (T)Tyrosine Tyr (Y)Tryptophan Trp (W)Valine Val (V)

[0071] It is understood that one way to define the variants and derivatives of the disclosed proteins herein is to define them in terms of homology / identity to specific known sequences. Specifically disclosed are vanants of peptides herein disclosed which have at least 70% or at least 75% or at least 80% or at least 85% or at least 90% or at least 95% or at least 96% or at least 97% or at least 98% or at least 99% identity to the peptides specifically recited herein. Those of skill in the art readily understand how to determine the identity of two proteins.

[0072] The polypeptides can be modified by either natural processes, such as post-translational processing, or by chemical modification techniques which are well known in the art. Modifications can occur anywhere in the polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. The same type of modification can be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide can have many types of modifications. Modifications include, without limitation, acetylation, acylation, ADP-ribosylation, amidation, covalent cross-linking or cyclization, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of a phosphytidylinositol, disulfide bond formation, demethylation, formation of cysteine or pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristolyation, oxidation, pegylation, proteolytic processing, phosphory lation, prenylation, racemization, selenoylation, sulfation, and transfer-RNA mediated addition of amino acids to protein such as arginylation. (See Proteins - Structure and Molecular Properties 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); Posttranslational Covalent Modification of Proteins, B. C. Johnson, Ed., Academic Press, New York, pp. 1-12 (1983)).

[0073] As used herein, the term “treat” or "treating" refers to partially or completely alleviating, ameliorating, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing incidence of one or more symptoms or features of a particularATTORNEY DOCKET NO.37759.0682P1disease, disorder, and / or condition. For example, "treating" osteoporosis, osteoarthritis and / or cartilage defects may refer to increasing tendon, cartilage or bone repair. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition and / or to a subject who exhibits only early signs of a disease, disorder, and / or condition for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. In some aspects, “to treat” can mean to administer a molecule or composition of the invention to a subject, such as a human or other mammal (for example, an animal model), that has an increased susceptibility for developing osteoporosis, osteoarthritis and / or cartilage defects, or that has osteoporosis, osteoarthritis and / or cartilage defects, in order to prevent or delay a worsening of the effects of the disease or condition, or to partially or fully reverse the effects of the disease.

[0074] As used herein, “osteochondral factor” refers to biological compounds that promote bone and / or cartilage repair and / or regeneration. Osteochondral factors include, but are not limited to, NELL-1, TGF-pi, TGF- 2, TGF- 3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects, osteochondral factors are derivatives of biological compounds that promote bone and / or cartilage repair and / or regeneration. In some aspects, osteochondral factors can be BMP2-K3Plk and BMP2-E83stop.

[0075] As used herein, “osteogenic factor” refers to biological compounds that promote new bone and / or new cartilage formation. Buser Z et al. Eur Spine J. 2017 Nov;26(l 1 ):2763-2772. Osteogenic factors include, but are not limited to, oxy 133 and oxysterol- 133.

[0076] As used herein, “tenogenic factor” refers to biological compounds that promote tendon repair and / or regeneration. Tenogenic factors include, but are not limited to TGF0-1, TGFP-2, TGF-P3, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1.

[0077] As used herein, “ECM protein” refers to proteins that are present in the extracellular matrix. The extracellular matrix is a three-dimensional network of extracellular macromolecules and minerals (including hydroxyapatite) that provide structural and biochemical support to surrounding cells. ECM proteins include, but are not limited to collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCNL WISP-1 or CTGF.

[0078] As used herein, an “agonist” is a molecule that binds to a target (e.g. a receptor) and activates the target in order to produce the target’s natural biological response. An agonist can competitively bind to the target in order to outcompete the target’s endogenous ligand. An agonist can also synergistically bind to a target in order to further amplify or increase the target’s natural biological response.ATTORNEY DOCKET NO.37759.0682P1

[0079] As used herein, an “antagonist'’ is a molecule that binds to a specific target (e.g., a receptor) and interferes with, inhibits or otherwise blocks activation of the target in order to inhibit the targets natural biological response.

[0080] As used herein, an “inhibitor” is a molecule that binds to a target (e.g., an enzyme or a receptor) in order to block, reduce, or stop the target’s biological response. For example, an inhibitor can competitively bind to an enzyme or a receptor in order to block, reduce, or stop the biological response.

[0081] As used herein, “tendon injury” refers to damage or inflammation of the connective tissue that links muscle to bone. It can be caused by overuse, degeneration, or trauma. As used herein, treating a tendon injury includes promoting soft tissue to bone healing.

[0082] As used herein, “fracture” refers to a break in a bone.

[0083] As used herein, “bone stress injury” refers to overuse injuries that are typified by a gradual onset of pain with activity. Examples of bone stress injuries include, but are not limited to, stress reactions and stress fractures. Bone stress injuries develop in response to repetitive loads applied to bone. Bone stress injuries are often seen on MRI as high signal intensity.

[0084] As used herein, treating “osteointegration of an implantable prosthesis” includes enhanced healing of bone to an implant material (metal, ceramic, etc.) when that implant is implanted into bone or dental applications.

[0085] As used herein, treating “osteointegration of allograft tissue” includes, but is not limited to, osteochondral allografts and bone allografts.

[0086] As used herein, “osteoarthritis” includes, but is not limited to, degenerative cartilage damage within a joint or intervertebral disc space, including damage from abnormal bone remodeling. Treatment of osteoarthritis includes, but is not limited to, treatment of osteoarthritic bone abnormalities, including subchondral bone and bone marrow edema as well as osteoarthritic cartilage abnormalities as the bone and cartilage abnormalities associated with osteoarthritis are related.

[0087] As used herein “cartilage defects” includes, but is not limited to, areas of damaged cartilage surrounded by relatively more healthy cartilage. In some aspects, the cartilage defect can be a defect or injury to articular or hyaline cartilage, fibrocartilage or elastic cartilage. The cartilage defects can be the result of traumatic mechanical destruction or progressive mechanical destructions. In some aspects, the cartilage defect can be in a specific, localized area of damage to the cartilage.ATTORNEY DOCKET NO.37759.0682P1

[0088] As used herein “osteoporosis” refers to a medical condition in which the bones become brittle and fragile from loss of tissue, typically as a result of hormonal changes, or deficiency of calcium or vitamin D.

[0089] As used herein “ectopic bone formation” refers to any bone growth outside of the normal skeleton. Thus, ectopic bone formation can refer to the process by which bone tissue forms outside of the skeleton in muscles and soft tissue.

[0090] As used herein “heterotopic ossification” refers to the presence of mature lamellar bone tissue outside of the bones that form the skeleton. In some aspects, heterotopic ossification can be a form of ectopic bone formation. In some aspects, heterotopic ossification and ectopic bone formation can even be used interchangeably.

[0091] As used herein “enthesis ossification at an injury’ site” refers to the transformation of soft tissue (tendon or ligament) into bone at its attachment point to the skeleton (the enthesis) following injury.

[0092] “Dose” or “dosage” as used herein refers to a specific quantity' of a therapeutic agent, such as a molecule, composition or pharmaceutical composition that is taken at specific times.

[0093] As used herein, “treat” is meant to mean administer one of the disclosed compositions to a subject, such as a human or other mammal (for example, an animal model), that has atherosclerosis, in order to prevent or delay a worsening of the effects of the disease or condition, or to partially or fully reverse the effects of the disease.

[0094] As used herein, “effective amount” is meant to mean a sufficient amount of one or more of the molecules, compositions or pharmaceutical compositions disclosed herein to provide the desired effect. For example, an effective amount of one or more of the molecules, compositions or pharmaceutical compositions disclosed herein can be an amount that provides a therapeutic effect and provides sustained therapeutic effects after withdrawal of the treatment. An effective amount of one or more of the molecules, compositions or pharmaceutical compositions disclosed herein can be an amount that is able to cause a benefit illustrated by, for example, promoting healing of tendon injuries, speeding up recovery time, improving biomechanical properties (such as strength or toughness) of the bone, cartilage and / or tendon, and / or improving tissue organization of bone, cartilage and / or tendon, as well as an amount that allows for a sustained therapeutic effect after withdrawal of the molecule, composition or pharmaceutical composition. The exact amount required will vary' from subject to subject, depending on the species, age, and general condition of the subject, the severity of disease (or underlying genetic defect) that is being treated, the particularATTORNEY DOCKET NO.37759.0682P1compound used, its mode of administration, and the like. Thus, it is not possible to specify an exact “effective amount." However, an appropriate “effective amount" may be determined by one of ordinary skill in the art using only routine experimentation.

[0095] The phrase “nucleic acid” as used herein refers to a naturally occurring or synthetic oligonucleotide or polynucleotide, whether DNA or RNA or DNA-RNA hybrid, single-stranded or double-stranded, sense or antisense, which is capable of hybridization to a complementary nucleic acid by Watson-Crick base-pairing. Nucleic acids of the invention can also include nucleotide analogs (e.g., BrdU), and non-phosphodiester internucleoside linkages (e.g., peptide nucleic acid (PNA) or thiodiester linkages). In particular, nucleic acids can include, without limitation, DNA, RNA, cDNA, gDNA, ssDNA, dsDNA or any combination thereof.

[0096] The terms “vector” or “construct” refer to a nucleic acid sequence capable of transporting into a cell another nucleic acid to which the vector sequence has been linked. The term "expression vector" includes any vector, (e.g., a plasmid, cosmid or phage chromosome) containing a gene construct in a form suitable for expression by a cell (e.g., linked to a transcriptional control element). " Plasmid" and "vector" are used interchangeably, as a plasmid is a commonly used form of vector. Moreover, the invention is intended to include other vectors which serve equivalent functions.

[0097] The term “expression vector” is herein to refer to vectors that are capable of directing the expression of genes to which they are operatively-linked. Common expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.Recombinant expression vectors can comprise a nucleic acid as disclosed herein in a form suitable for expression of the acid in a host cell. In other words, the recombinant expression vectors can include one or more regulatory elements or promoters, which can be selected based on the host cells used for expression that is operatively linked to the nucleic acid sequence to be expressed.

[0098] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of skill in the art to which the disclosed method and compositions belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present method and compositions, the particularly useful methods, devices, and materials are as described.Publications cited herein and the material for which they are cited are hereby specifically incorporated by reference. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention. NoATTORNEY DOCKET NO.37759.0682P1admission is made that any reference constitutes prior art. The discussion of references states what their authors assert, and applicants reserve the right to challenge the accuracy and pertinency of the cited documents. It will be clearly understood that, although a number of publications are referred to herein, such reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art.B. Compositions1. Molecules

[0099] Non-limiting examples of the molecules of the disclosure are disclosed herein.

[0100] Disclosed herein are molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the bioactive moiety to the BP moiety.

[0101] Disclosed herein are molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety' has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety. Disclosed herein are molecules comprising: i) a piezol agonist, an IL- 1(3 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an Activin A receptor like type 1 (ALK1) inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL- 1(3 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor. NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.i. Bioactive Moiety

[0102] As used herein, “bioactive moiety” is meant a part of a molecule that has or can elicit an action in the body of a subject or a physiological or pharmacological response in the body of a subject.

[0103] In some aspects, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1(3 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK.1 inhibitor. In some aspects, the ECM protein can be, but is not limited to, collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0104] In some aspects, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be, but is not limited to, NELL-1, TGF-(31, TGF-(32, TGF-|33. BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11,ATTORNEY DOCKET NO.37759.0682P1BMP-15, or rhBMP. In some aspects, the osteochondral factor can be NELL-1 or BMP-2.

[0105] In some aspects, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedi 1 / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects, the bioactive moiety can be, but is not limited to, an IL-ip antagonist In some aspects, IL-ip antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects, the bioactive moiety can be an IRAK4 inhibitor. In some aspects, the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects, the bioactive moiety can be a caspase- 1 inhibitor. In some aspects, the caspase- 1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects, the bioactive moiety can be aNLRP3 inflammasome inhibitor. In some aspects, theNLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects, the bioactive moiety can be a salt-inducible kinase inhibitor. In some aspects, the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects, the bioactive moiety can be an Activin A receptor like type 1 (ALK1 ) inhibitor. In some aspects, theALKl inhibitor can be, but is not limited to LDN214117.

[0106] In some aspects, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, or an Extracellular Matrix (ECM) protein that has been genetic code expanded to comprise a non-natural amino acid.

[0107] In some aspects, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects, the variant of BMP -2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects, the variant of BMP -2 can be BMP2-K3Plk. In some aspects, the bioactive moiety can be BMP2-K3Plk. In some aspects, the variant of BMP-2 can be BMP2-E83stop. In some aspects, the bioactive moiety can be BMP2-E83stop. In some aspects, the variant of BMP-2 can be BMP2-A2C. In some aspects, the bioactive moiety can be BMP2-A2C. In some aspects, the variant of BMP -2 can be BMP2-E83Plk. In some aspects, the bioactive moiety can be BMP2-E83Plk. In some aspects, the variant of BMP-2 can be BMP2-E83Azide. In some aspects, the bioactive moiety can be BMP2-E83Azide. In some aspects, the variant of BMP-2 can be BMP2 E94Plk. In someATTORNEY DOCKET NO.37759.0682P1aspects, the bioactive moiety can be BMP2 E94Plk.

[0108] In some aspects, BMP2-K3Plk sequence is modified from the native BMP2 sequence such that the codon (AAA) coding for Lys3 (amino acid at position 3 of the mature portion of BMP2 protein) in the wild-type sequence was replaced by TAG (amber-, stopcodon). In some aspects, BMP2-K3PLK has propargyl-L-lysine at the 3rdamino acid in the BMP2 protein sequence. In some aspects, the TAG stop codon terminating the mature part of the wild-type BMP2 sequence is replaced by a TAA stop codon.

[0109] In some aspects, pyrrolysyl-tRNA synthethase, a specialized tRNA for the translation of propargyl-L-lysine, is used for the incorporation of propargyl-L-lysine into the BMP2 amino acid sequence.

[0110] In some aspects,L‘BMP2-E83stop” sequence is human BMP2 sequence where the codon (GAA) encoding for glutamic acid 83rdamino acid is replaced by the amber stopcodon TAG. In some aspects, BMP2-E83stop has H-L-Lys(EO-N3)-OH (Azide) at the 83rdamino acid in the BMP2 protein sequence.

[0111] In some aspects, BMP2-A2C sequence is modified from the native BMP2 sequence such that the sequence contains an additional cysteine residue by replacing alanine at the 2ndamino acid in the BMP2 protein sequence.

[0112] In some aspects, BMP2-E83Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 83 is replaced by propargyl-L-lysine (Plk).

[0113] In some aspects, BMP2-E83Azide sequence is modified from the native BMP2 sequence such that an azide-functionalized amino acid H-L-Lys(EO-N3)-OH (Azide) is present at the 83rdamino acid.

[0114] In some aspects, BMP2-E94Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 94 is replaced with propargyl-L-lysine (Plk).

[0115] In some aspects, the variant of BMP-2 can be made using the methods described herein and in the Example.Table 3SEQ ID NO: SequenceSEQ ID NO: 6 MAQA*HKQRKRLKSSCKRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLA (BMP2-K3Plk DHLNSTNHAI VQTL VN S VNSKIPKACC VPTEL SAI SML YLDENEK V VLKN YQDMV AA sequence) VEGCGCR*SEQ ID NO: 7 TGGCTCAAGCCTAGCACAAACAGCGGAAACGCCTTAAGTCCAGCTGTAAGAGA (BMP2-K3Plk CACCCTTTGTACGTGGACTTCAGTGACGTGGGGTGGAATGACTGGATTGTGGCT nucleic acid CCCCCGGGGTATCACGCCTTTTACTGCCACGGAGAATGCCCTTTTCCTCTGGCTsequence) GATCATCTGAACTCCACTAATCATGCCATTGTTCAGACGTTGGTCAACTCTGTTATTORNEY DOCKET NO.37759.0682P1AACTCTAAGATTCCTAAGGCATGCTGTGTCCCGACAGAACTCAGTGCTATCTCG ATGCTGTACCTTGACGAGAATGAAAAGGTTGTATTAAAGAACTATCAGGACAT GGTTGTGGAGGGTTGTGGGTGTCGCTAA SEQ ID NO: 8 vector sequence Ndel amber stopPyrrolysyl- 5 ' catATG GCT C AA GCC TAG AAA CAC AAA CAG CGG AAA CGC tRNA CTT AAG TCC AGC TGT AAG AGA CAC CCT TTG TAC GTG GAC TTC AGT GAC synthethase GTG GGG TGG AAT GAC TGG ATT GTG GCT CCC CCG GGG TAT CAC GCC TTT gene sequence: TAC TGC CAC GGA GAA TGC CCT TTT CCT CTG GCT GAT CAT CTG AAC TCC ACT AAT CAT GCC ATT GTT CAG ACG TTG GTC AAC TCT GTT AAC TCT AAG ATT CCT AAG GCA TGC TGT GTC CCG ACA GAA CTC AGT GCT ATC TCG ATG CTG TAC CTT GAC GAG AAT GAA AAG GTT GTA TTA AAG AAC TAT CAG GAC ATG GTT GTG GAG GGT TGT GGG TGT CGC TAG TAA GGA TCC GCA AAA GCG GCC TTT GAC TCC CTG CAA GCC TCA GCG ACC GAA TAT ATC GGT TAT GCG TGG GCG ATG GTT GTT GTC ATT GTC GGC GCA ACT ATC GGT ATC AAG CTG I TT AAG AAA T TC ACC TCG AAA GCA AGC TAA ggatcc. 3'stop BamHI Vector sequence

[0116] In some aspects, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can be, but not limited to, oxy133 or oxysterol-133.

[0117] In some aspects, the bioactive moiety can be atenogenic factor, and the tenogenic factor can be, but not limited to, TGF0-1, TGF0-2, TGF-03, FGF, CTGF, BMP-12, BMP-13, BMP-14. CCN1 or WISP-1. In some aspects, the tenogenic factor can be TGF0-1 or TGF0-2.

[0118] In some aspects, the bioactive moiety can be a hormone. In some aspects, the hormone can be parathyroid hormone (e.g., teriparatide, Natpara). In some aspects, the bioactive moiety can be parathyroid hormone-related protein.

[0119] In some aspects, the bioactive moiety can be a growth differentiation factor. In some aspects, the grow th differentiation factor can be growth differentiation factor 11 (GDF11) or bone morphogenic protein 11 (BMP-11).

[0120] In some aspects, the bioactive moiety' can be activin receptor-like kinase- 1-Fc (ALKl-Fc, dalantercept). ALKl-Fc is a chimeric protein with ALK1 receptor domain combined with the Fc portion of human IgG and serves as a ligand trap for ALK1 ligands BMP9 and BMP 10.

[0121] In some aspects, the bioactive moiety' can be activin receptor-like kinase-4-Fc (ALK4-Fc). ALK4-Fc is a chimeric protein with ALK4 receptor domain combined with the Fc portion of human IgG and serves as a ligand trap for ALK4 ligands activins, GDF8 and GDF11.

[0122] In some aspects, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ATTORNEY DOCKET NO.37759.0682P1ii. Cleavable Linker

[0123] As used herein, “cleavable linker’ is meant a peptide linker that is capable of being cleaved. In some aspects, the cleavable linker can be enzymatically cleaved (i.e. enzymatically cleavable linkers). In some aspects, the cleavable linker can be a proteasesensitive peptide linker, acid sensitive hydrazone linker, a glutathione-sensitive disulfide linker, or a photocleavable linker. Examples of cleavable linkers include, but are not limited to peptides which are capable of being cleaved by the enzyme cathepsin K (cathepsin K-sensitive linker) or by the matrix metalloproteinase-2 enzyme (MMP linker). Cleavable linkers can also include, but are not limited to, peptide linkers that are substrates for other matrixmetalloproteinase enzymes such as MMP-1, MMP-3, MMP-9, MMP-10 and MMP-13. Examples of cleavable linkers can include, but are not limited to the cleavable linkers in Table 4.Table 4 - Examples of Cleavable LinkersEnzyme Amino acid sequence SEQ ID NO:Cathepsin K (Ctsk) GHPGGPQG 1Cathepsin K (Ctsk) GGGMGPSGPWGGK 2MMP PLGLAG 3MMP GCRD-GPQGIWGQ-DRCG 4MMP GCRD-GPQGIAGQ-DRCG 5MMP SGESPAYYTA 9MMP GAPFALRLV 10MMP GGYAELRMGG 11MMP GPLGLWAR 12MMP GPLGMRGL 13MMP IPESLRAG 14MMP PVG↓LIG (PVGLIG) 15MMP PLG↓LAG (PLGLAG) 16MMP PLGVR 17MMP VPMSMRGG 18MMP GPQG↓IWGQ (GPQGIWGQ) 19MMP GPQG↓IAGQ (GPQGIAGQ) 20Cathepsin K (Ctsk) HPGGPQ 21Cathepsin K (Ctsk) KPRGSKQ 22ATTORNEY DOCKET NO.37759.0682P1Cathepsin K (Ctsk) KKPGSKQ 23Cathepsin K (Ctsk) Z-GPR-AMC** Z-GPR-AMC = Z=Cbz=Benzyloxycarbonyl; AMC=7-amino-4-methylcoumarin

[0124] Also disclosed a variants or derivatives of the cleavable linkers disclosed herein. As used herein, the term “analog” is used interchangeably with “variant” and “derivative.” Variants and derivatives are well understood to those of skill in the art and can involve amino acid sequence modifications. Such amino acid sequence modifications typically fall into one or more of three classes: substantial; insertional; or deletional variants. Insertions include amino and / or carboxyl terminal fusions as well as intrasequence insertions of single or multiple amino acid residues. Insertions ordinarily are smaller insertions than those of amino or carboxyl terminal fusions, for example, on the order of one to four residues. These variants ordinarily are prepared by site-specific mutagenesis of nucleotides in the DNA encoding the protein, thereby producing DNA encoding the variant, and thereafter expressing the DNA in recombinant cell culture. Techniques for making substitution mutations at predetermined sites in DNA having a known sequence are well known, for example Ml 3 primer mutagenesis and PCR mutagenesis. Amino acid substitutions are typically of single residues, but can occur at a number of different locations at once. Substitutions, deletions, insertions or any combination thereof may be combined to arrive at a final derivative or analog.Substitutional variants are those in which at least one residue has been removed and a different residue inserted in its place. Such substitutions generally are made in accordance with Tables 1 and 2 and are referred to as conservative substitutions.

[0125] In some aspects, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0126] In some aspects, the cleavable linker can be an MMP cleavable linker. In some aspects, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ IDATTORNEY DOCKET NO.37759.0682P1NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0127] In some aspects, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent Nos. 8,063,209, 11,219,697 and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent Nos. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0128] As used herein, “bisphosphonate moiety’’ (BP moiety) refers to a molecule characterized by two C P bonds. When the two C P bonds share a single carbon atom (P-C-P), they are deemed to be analogues of pyrophosphate (P-O-P) and are called geminal bisphosphonates (so-called because the carbon is at the central or geminal position). The P-C-P bonds of the geminal bisphosphonate are stable to heat and most chemical reagents and are completely resistant to enzymatic hydrolysis. In some aspects, a “bisphosphonate moiety" refers to a molecule having two phosphate ions connected by a carbon atom (a P-C-P structure). Bisphosphonates are analogues of pyrophosphate that contain a carbon instead of an oxygen atom. The single P-C-P structure can allow a great number of possible variations, especially by changing the two lateral chains on the carbon atom. In some aspects, a “bisphosphonate moiety’’ exhibits a high affinity for exposed hydroxyapatite (HAP) calciumphosphate mineral in the ECM of bone and will preferentially bind newly resorbed bone surfaces. Some bisphosphonates, such as etidronate and methylhydroxyl diphosphonate, do not inhibit or weakly inhibit the function of osteoclasts. In general, nitrogen-containing bisphosphonates as a class of anti-osteoporosis drugs inhibit osteoclasts more strongly than non-nitrogen containing bisphosphonates. See USPN 11,400,104 and Hokugo A et al. Bone.2019 Jun;123: 115-128. In some aspects, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects, the BP may include nitrogencontaining bisphosphonates in which the nitrogen basicity is greatly decreased by derivatization of the amine group, e.g. to form an acid amide. In some aspects, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2- phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, pamidronate or zolendronate. In some aspects, the BP moiety has an aminoalkyl side chain and H or OH asATTORNEY DOCKET NO.37759.0682P1the other P-C-P carbon substituent. In some aspects, the BP moiety can be alendronate, pamidronate. or zolendronate. In some aspects, the BP moiety’ can be alendronate. In some aspects, the BP moiety can be zolendronate. In some aspects, the BP moiety can be pamidronate.

[0129] In some aspects, the chemical name for pamidronic acid is (3 -amino- 1 -hy droxy- 1-phosphonopropyl)phosphonic acid. In some aspects, the chemical name for alendronic acid is (4-amino- 1 -hydroxy- 1 -phosphonobutyl)phosphonic acid.iv. Spacer

[0130] In some aspects, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety' or the cleavable linker and the bisphosphonate moiety. In some aspects, the spacer can be a peptide spacer. In some aspects, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62. 63. 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137. 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155. 156, 157, 158. 159, 160. 161, 162, 163, 164, 165. 166, 167. 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227. 228, 229, 230, 231, 232. 233, 234, 235, 236, 237, 238, 239. 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility', improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0131] In some aspects, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety' or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0132] In some aspects, the spacer can be Beta-alanine, GABA, AEA, Ava, Ahx, PEG2ATTORNEY DOCKET NO. 37759.0682P1Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0133] Disclosed herein are molecules comprising the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityN–Gly–His–Pro–Gly–Gly–Pro–Gln–GlyPEPTIDE LINKERCtsk-substrate for cleavage

[0134] Disclosed herein are molecules comprising the structure:2. Compositions

[0135] Disclosed herein are compositions comprising i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has aP-C-P structure and wherein the cleavable linker links the bioactive moiety to the BP moiety. Disclosed herein are compositions comprising i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP -2 to the BP moiety. Disclosed herein are compositions comprising: i) a piezol agonist, an IL-1 (3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, an IL- ip antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.i. Bioactive Moiety

[0136] In some aspects, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-ip antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasomeATTORNEY DOCKET NO.37759.0682P1inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects, the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0137] In some aspects, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be, but is not limited to, NELL-1, TGF-01, TGF-P2, TGF-(33, BMP-1, BMP-2, BMP-3. BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b. BMP-10. BMP-11, BMP-15, or rhBMP. In some aspects, the osteochondral factor can be NELL-1 or BMP-2.

[0138] In some aspects, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects, the bioactive moiety can be, but is not limited to, an IL- 1 P antagonist In some aspects, IL- 1 P antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects, the bioactive moiety can be an IRAK4 inhibitor. In some aspects, the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects, the bioactive moiety can be a caspase- 1 inhibitor. In some aspects, the caspase- 1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects, the bioactive moiety can be a NLRP3 inflammasome inhibitor. In some aspects, theNLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects, the bioactive moiety can be a salt-inducible kinase inhibitor. In some aspects, the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRI A9, OMX-0407, Pterosin B, KIN- 112, HG-9-91 -01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor. In some aspects, the ALK1 inhibitor can be, but is not limited to LDN214117.

[0139] In some aspects, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0140] In some aspects, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be a variant of BMP-2. In some aspects, the variant of BMP -2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects, the variant of BMP -2 can be BMP2-K3Plk. In some aspects, the bioactive moiety can be BMP2-K3Plk. In some aspects.ATTORNEY DOCKET NO.37759.0682P1the variant of BMP-2 can be BMP2-E83stop. In some aspects, the bioactive moiety can be BMP2-E83stop. In some aspects, the variant of BMP-2 can be BMP2-A2C. In some aspects, the bioactive moiety can be BMP2-A2C. In some aspects, the variant of BMP-2 can be BMP2-E83Plk. In some aspects, the bioactive moiety can be BMP2-E83Plk. In some aspects, the variant of BMP-2 can be BMP2-E83Azide. In some aspects, the bioactive moiety can be BMP2-E83Azide. In some aspects, the variant of BMP-2 can be BMP2 E94Plk. In some aspects, the bioactive moiety can be BMP2 E94Plk.

[0141] In some aspects, BMP2-K3Plk sequence is modified from the native BMP2 sequence such that the codon (AAA) coding for Lys3 (amino acid at position 3 of the mature portion of BMP2 protein) in the wild-type sequence was replaced by TAG (amber-, stopcodon). In some aspects. BMP2-K3PLK has propargyl-L-lysine at the 3rd amino acid in the BMP2 protein sequence. In some aspects, the TAG stop codon terminating the mature part of the wild-type BMP2 sequence is replaced by a TAA stop codon.

[0142] In some aspects, pyrrolysyl-tRNA synthethase, a specialized tRNA for the translation of propargyl-L-lysine, is used to for the incorporation of propargyl-L-lysine into the BMP2 amino acid sequence.

[0143] In some aspects, “BMP2-E83stop” sequence is human BMP2 sequence where the codon (GAA) encoding for glutamic acid 83rdamino acid is replaced by the amber stopcodon TAG. In some aspects, BMP2-E83stop has H-L-Lys(EO-N3)-OH (Azide) at the 83rdamino acid in the BMP2 protein sequence.

[0144] In some aspects, BMP2-A2C sequence is modified from the native BMP2 sequence such that the sequence contains an additional cysteine residue by replacing alanine at the 2ndamino acid in the BMP2 protein sequence.

[0145] In some aspects, BMP2-E83Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 83 is replaced by propargyl-L-lysine (Plk).

[0146] In some aspects, BMP2-E83Azide sequence is modified from the native BMP2 sequence such that an azide-functionalized amino acid H-L-Lys(EO-N₃)-OH (Azide) is present at the 83rdamino acid.

[0147] In some aspects, BMP2 E94Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 94 is replaced with propargyl-L-lysine (Plk).

[0148] In some aspects, the variant of BMP-2 can be made using the methods described herein and in the Examples.ATTORNEY DOCKET NO.37759.0682P1

[0149] In some aspects, the bioactive moiety' can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0150] In some aspects, the bioactive moiety can be atenogenic factor, and the tenogenic factor can be, but is not limited to, TGF|3-1, TGF0-2, TGF-03, FGF, CTGF, BMP- 12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects, the tenogenic factor can be TGF0-1 or TGFP-2.

[0151] In some aspects, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0152] In some aspects, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0153] In some aspects, the cleavable linker can be an MMP cleavable linker. In some aspects, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11 ) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0154] In some aspects, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent Nos. 8,063,209, 11,219,697 and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent Nos. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0155] In some aspects, the BP moiety' does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects, the BP moiety can be. but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-ATTORNEY DOCKET NO.37759.0682P1(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects, the BP moiety can be alendronate. In some aspects, the BP moiety can be zolendronate. In some aspects, the BP moiety can be pamidronate.iv. Spacer

[0156] In some aspects, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety7or the cleavable linker and the bisphosphonate moiety. In some aspects, the spacer can be a peptide spacer. In some aspects, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87. 88. 89. 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155. 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167. 168, 169, 170, 171, 172, 173. 174, 175, 176. 177, 178. 179, 180, 181, 182, 183. 184, 185. 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245. 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility7and increase flexibility, improve the biological activity7of the molecule or to improve the stability7and bioavailability of the molecule.

[0157] In some aspects, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety ). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0158] In some aspects, the spacer can be. but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.ATTORNEY DOCKET NO.37759.0682P1

[0159] In some aspects, the spacer can be an azide spacer. In some aspects, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0160] Disclosed herein are compositions comprising molecules comprising the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityPro - Gly -Giy -Pro • Gin - Gly 'zPEPTIDE LINKERCtsk-substrate for cleavage

[0161] Disclosed herein are compositions comprising molecules comprising the structure:3. Pharmaceutical Compositions

[0162] Disclosed herein are pharmaceutical compositions comprising i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the bioactive moiety to the BP moiety. Disclosed herein are pharmaceutical compositions comprising i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety- has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.Disclosed herein are pharmaceutical compositions comprising: i) a piezol agonist, an IL-10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK.1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL-10 antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.i. Bioactive Moiety

[0163] In some aspects, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasomeATTORNEY DOCKET NO.37759.0682P1inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects, the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0164] In some aspects, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-01, TGF-P2, TGF-(33, BMP-1, BMP-2, BMP-3. BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b. BMP-10. BMP-11, BMP-15, or rhBMP. In some aspects, the osteochondral factor can be NELL-1 or BMP-2.

[0165] In some aspects, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects, the bioactive moiety can be, but is not limited to, an IL- 1 P antagonist In some aspects, IL- 1 P antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects, the bioactive moiety can be an IRAK4 inhibitor. In some aspects, the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects, the bioactive moiety can be a caspase- 1 inhibitor. In some aspects, the caspase- 1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects, the bioactive moiety can be a NLRP3 inflammasome inhibitor. In some aspects, theNLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects, the bioactive moiety can be a salt-inducible kinase inhibitor. In some aspects, the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124. ARN-3236, GLPG3312, MRI A9, OMX-0407, Pterosin B, KIN- 112, HG-9-91 -01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor. In some aspects, the ALK1 inhibitor can be, but is not limited to LDN214117.

[0166] In some aspects, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0167] In In some aspects, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects, the variant of BMP -2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects, the variant of BMP-2 can be BMP2-K3Plk. In some aspects, the bioactive moiety can be BMP2-K3Plk. In some aspects.ATTORNEY DOCKET NO.37759.0682P1the variant of BMP-2 can be BMP2-E83stop. In some aspects, the bioactive moiety can be BMP2-E83stop. In some aspects, the variant of BMP-2 can be BMP2-A2C. In some aspects, the bioactive moiety can be BMP2-A2C. In some aspects, the variant of BMP-2 can be BMP2-E83Plk. In some aspects, the bioactive moiety can be BMP2-E83Plk. In some aspects, the variant of BMP-2 can be BMP2-E83Azide. In some aspects, the bioactive moiety can be BMP2-E83Azide. In some aspects, the variant of BMP-2 can be BMP2 E94Plk. In some aspects, the bioactive moiety can be BMP2 E94Plk.

[0168] In some aspects, BMP2-K3Plk sequence is modified from the native BMP2 sequence such that the codon (AAA) coding for Lys3 (amino acid at position 3 of the mature portion of BMP2 protein) in the wild-type sequence was replaced by TAG (amber-, stopcodon). In some aspects. BMP2-K3PLK has propargyl-L-lysine at the 3rd amino acid in the BMP2 protein sequence. In some aspects, the TAG stop codon terminating the mature part of the wild-type BMP2 sequence is replaced by a TAA stop codon.

[0169] In some aspects, pyrrolysyl-tRNA synthethase, a specialized tRNA for the translation of propargyl-L-lysine, is used to for the incorporation of propargyl-L-lysine into the BMP2 amino acid sequence.

[0170] In some aspects, “BMP2-E83stop” sequence is human BMP2 sequence where the codon (GAA) encoding for glutamic acid 83rdamino acid is replaced by the amber stopcodon TAG. In some aspects, BMP2-E83stop has H-L-Lys(EO-N3)-OH (Azide) at the 83rdamino acid in the BMP2 protein sequence.

[0171] In some aspects, BMP2-A2C sequence is modified from the native BMP2 sequence such that the sequence contains an additional cysteine residue by replacing alanine at the 2ndamino acid in the BMP2 protein sequence.

[0172] In some aspects, BMP2-E83Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 83 is replaced by propargyl-L-lysine (Plk).

[0173] In some aspects, BMP2-E83Azide sequence is modified from the native BMP2 sequence such that an azide-functionalized amino acid H-L-Lys(EO-N₃)-OH (Azide) is present at the 83rdamino acid.

[0174] In some aspects, BMP2 E94Plk sequence is modified from the native BMP2 sequence such that the glutamic acid (E) at position 94 is replaced with propargyl-L-lysine (Plk).

[0175] In some aspects, the variant of BMP-2 can be made using the methods described herein and in the Examples.ATTORNEY DOCKET NO.37759.0682P1

[0176] In some aspects, the bioactive moiety' can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0177] In some aspects, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF|3-1, TGF0-2, TGF-03, FGF, CTGF, BMP- 12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects, the tenogenic factor can be TGFp-1 or TGFP-2.

[0178] In some aspects, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0179] In some aspects, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0180] In some aspects, the cleavable linker can be an MMP cleavable linker. In some aspects, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11 ) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0181] In some aspects, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent Nos. 8,063,209, 11,219,697 and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent Nos. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0182] In some aspects, the BP moiety' does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects, the BP moiety can be. but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-ATTORNEY DOCKET NO.37759.0682P1(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects, the BP moiety can be alendronate. In some aspects, the BP moiety can be zolendronate. In some aspects, the BP moiety can be pamidronate.iv. Spacer

[0183] In some aspects, the pharmaceutical compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety7or the cleavable linker and the bisphosphonate moiety. In some aspects, the spacer can be a peptide spacer. In some aspects, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84. 85. 86. 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153. 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165. 166, 167, 168, 169, 170, 171. 172, 173, 174. 175, 176. 177, 178, 179, 180, 181. 182, 183. 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243. 244, 245, 246, 247, 248. 249, or 250 amino acids, or any derivable range therein. In some aspects, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility7and increase flexibility, improve the biological activity7of the molecule or to improve the stability7and bioavailability of the molecule.

[0184] In some aspects, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety ). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0185] In some aspects, the spacer can be. but is not limited, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.ATTORNEY DOCKET NO. 37759.0682P1

[0186] In some aspects, the spacer can be an azide spacer. In some aspects, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0187] Disclosed herein are pharmaceutical compositions comprising molecules comprising the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinity•Gly His Pre Gly Gly -Pro • Gin - GlyPEPTIDE LINKERCtsk-substrate for cleavage

[0188] Disclosed herein are pharmaceutical compositions comprising molecules comprising the structure:

[0189] By “pharmaceutically acceptable” is meant a material or carrier that would be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject, as would be well known to one of skill in the art. Examples of carriers include dimyristoylphosphatidyl (DMPC), phosphate buffered saline or a multivesicular liposome. For example, PG: PC: Cholesterol:peptide or PC:peptide can be used as carriers in this invention. Other suitable pharmaceutically acceptable carriers and their formulations are described in Remington: The Science and Practice of Pharmacy (19th ed.) ed. A. R. Gennaro, Mack Publishing Company, Easton, PA 1995. Typically, an appropriate amount of pharmaceutically-acceptable salt is used in the formulation to render the formulation isotonic. Other examples of the pharmaceutically-acceptable carrier include, but are not limited to, saline, Ringer’s solution and dextrose solution. The pH of the solution can be from about 5 to about 8, or from about 7 to about 7.5. Further carriers include sustained release preparations such as semi-permeable matrices of solid hydrophobic polymers containing the composition, which matrices are in the form of shaped articles, e.g., films, stents (which are implanted in vessels during an angioplasty procedure), liposomes or microparticles. It will be apparent to those persons skilled in the art that certain carriers may¬ be more preferable depending upon, for instance, the route of administration andATTORNEY DOCKET NO.37759.0682P1concentration of composition being administered. These most typically would be standard carriers for administration of drugs to humans, including solutions such as sterile water, saline, and buffered solutions at physiological pH.

[0190] Pharmaceutical compositions can also include carriers, thickeners, diluents, buffers, preservatives and the like, as long as the intended activity' of the molecule, composition or pharmaceutical compositions of the invention is not compromised.Pharmaceutical compositions may also include one or more active ingredients (in addition to the composition of the invention) such as antimicrobial agents, anti-inflammatory agents, anesthetics, and the like. The pharmaceutical composition may be administered in a number of ways depending on whether local or systemic treatment is desired, and on the area to be treated.

[0191] Preparations of parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer’s dextrose, dextrose and sodium chloride, lactated Ringer’s, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer’s dextrose), and the like. Preservatives and other additives may also be present such as. for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.

[0192] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids, or binders may be desirable. Some of the compositions may potentially be administered as a pharmaceutically acceptable acid- or base- addition salt, formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases such as mon-, di-, trialkyl and aryl amines and substituted ethanolamines. C. Methods1. Methods For Targeting a Molecule to a Site of Injury

[0193] Disclosed herein are methods of targeting a molecule to a site of injury in aATTORNEY DOCKET NO.37759.0682P1subject in need thereof comprising administering any one of the disclosed molecules, compositions or pharmaceutical compositions to the subject.

[0194] Disclosed herein are methods of targeting a molecule to a site of injury in a subject in need thereof comprising administering a molecule comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, a piezol agonist, an IL-1 p antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0195] Disclosed herein are methods of targeting a molecule to a site of injury in a subject in need thereof comprising administering a molecule comprising: i) a variant of BMP -2, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0196] Disclosed herein are methods of targeting a molecule to a site of injury in a subject in need thereof comprising administering molecules comprising: i) a piezol agonist, an IL-ip antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK.1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL-1 antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0197] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the site of injury is enthesis tissue. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the site of injury can be, but is not limited to, a rotator cuff, a distal biceps tendon, a pectoralis major, a patellar tendon, a quadriceps tendon, or a triceps tendon.

[0198] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the site of injury can be a site of surgery' and the surgery can be. but is not limited to, an ACL reconstruction, a PCL reconstruction, a LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, or an osteochondral allograft transplantation.ATTORNEY DOCKET NO.37759.0682P1

[0199] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0200] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, atenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety is an ECM protein and the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0201] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11. BMP-15, or rhBMP. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP-2.

[0202] In some aspects, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects, the bioactive moiety can be, but is not limited to, an IL- 10 antagonist In some aspects, IL- 10 antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects, the bioactive moiety can be an IRAK4 inhibitor. In some aspects, the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects, the bioactive moiety can be a caspase- 1 inhibitor. In some aspects,the caspase- 1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740ATTORNEY DOCKET NO.37759.0682P1(pralnacasan). In some aspects, the bioactive moiety can be a NLRP3 inflammasome inhibitor. In some aspects, theNLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects, the bioactive moiety can be a salt-inducible kinase inhibitor. In some aspects, the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects, the bioactive moiety can be an Activin A receptor like type 1 (ALK1 ) inhibitor. In some aspects, theALKl inhibitor can be, but is not limited to LDN214117.

[0203] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0204] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-K3Plk. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-E83stop. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods ofATTORNEY DOCKET NO.37759.0682P1targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-E83Azide. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0205] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the variant of BMP -2 can be made using the methods described herein and in the Examples.

[0206] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0207] In some aspects of the methods of targeting a molecule to a site of injury7in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF0-1. TGF0-2. TGF-03, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects ofthe methods of targeting a molecule to a site of injury' in a subject in need thereof disclosed herein, the tenogenic factor can be TGF0-1 or TGF0-2.

[0208] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0209] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQATTORNEY DOCKET NO.37759.0682P1ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0210] In some aspects of the methods of targeting a molecule to a site of injury7in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0211] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697, and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0212] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of targeting a molecule to a site of injury7in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-(pyridin-4-yl)ethane-I,l-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspectsATTORNEY DOCKET NO.37759.0682P1of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0213] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety7. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 ammo acids, or any derivable range therein. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0214] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited toATTORNEY DOCKET NO.37759.0682P1hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0215] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Betaalanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0216] In some aspects of the methods of targeting a molecule to a site of injury7in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group. In some aspects of the methods of targeting a molecule to a site of injury' in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityN-Gly-His-Pro-Gly-Gly-Pro-Gln-GlyPEPTIDE LINKERCtsk-substrate for cleavage

[0217] In some aspects of the methods of targeting a molecule to a site of injury in a subject in need thereof disclosed herein, the molecules comprise the structure:2. Methods of Treating a Tendon Injury and / or Promoting Soft Tissue to Bone Healing

[0218] Disclosed are methods of treating a tendon injury in a subject in need thereof comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject. Also disclosed are methods of promoting soft tissue to bone healing comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject.

[0219] Disclosed herein methods of treating a tendon injury in a subject in need thereof comprising administering to the subject molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-PATTORNEY DOCKET NO.37759.0682P1structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0220] Disclosed herein are methods of treating a tendon injury in a subject in need thereof comprising administering to the subject molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0221] Disclosed herein are methods of treating a tendon injury in a subject in need thereof comprising administering to the subject molecules comprising: i) a piezol agonist, an IL-10 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist. IL- 10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0222] In some aspects of the methods of treating a tendon injury' in a subject in need thereof disclosed herein, the tendon injury can be a rotator cuff, a distal biceps tendon, pectoralis major, patellar tendon, quadriceps tendon, or a triceps tendon.

[0223] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of treating a tendon injury' in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0224] In some aspects of the methods of treating a tendon injury^ in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL- 10 antagonist, an IRAK.4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK.1 inhibitor. In some aspects of the methods ofATTORNEY DOCKET NO.37759.0682P1treating a tendon injur)- in a subject in need thereof disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0225] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-(31, TGF-(32, TGF-(33. BMP-1, BMP-2, BMP-3. BMP-4, BMP-5. BMP-6, BMP-7. BMP-8a. BMP-8b. BMP-10. BMP-11, BMP-15, or rhBMP. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP -2.

[0226] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (K. C289), KC159, Yaddlel, Jedi 1 / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an IL-1 (3 antagonist, and the IL- 113 antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a caspase-1 inhibitor, and the caspase- 1 inhibitor can be, but is not limited to. VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061. YKL-06-062, Dasatinib. Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0227] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise aATTORNEY DOCKET NO.37759.0682P1non-natural amino acid. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0228] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-K3Plk. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-E83stop. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety7can be BMP2-E83stop. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of treating a tendon injury7in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of treating a tendon injury7in a subject in need thereof disclosed herein, the bioactive moiety7can be BMP2 E94Plk.

[0229] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be made using the methods described herein and in the Examples.ATTORNEY DOCKET NO.37759.0682P1

[0230] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0231] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF0-1, TGF0-2, TGF-03, FGF, CTGF, BMP- 12, BMP-13, BMP- 14, CCN1 or WISP-1. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the tenogenic factor can be TGF0-1 or TGF0-2.

[0232] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11). bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0233] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0234] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0235] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitiveATTORNEY DOCKET NO.37759.0682P1linkers can be those found in US Patent No. 8,063,209, 11,219,697, and US 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0236] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-(pyridin-4-yl)ethane-I, I-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of treating a tendon injury7in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0237] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of treating a tendon injury7in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100,101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118,ATTORNEY DOCKET NO. 37759.0682P1119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138. 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150. 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210. 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222. 223, 224, 225, 226, 227, 228. 229, 230, 231. 232, 233, 234, 235, 236, 237, 238. 239, 240. 241, 242, 243. 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility’ and increase flexibility, improve the biological activity7of the molecule or to improve the stability’ and bioavailability of the molecule.

[0238] In some aspects of the methods of treating a tendon injury’ in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0239] In some aspects of the methods of treating a tendon injury’ in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava. Ahx. PEG2 Spacer, PEG3 Spacer. PEG4 Spacer, or Ttds.

[0240] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0241] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityN-Gly-His-Pro-Gly-Gly-Pro-Gln-GlyPEPTIDE LINKERCtsk- substrate for cleavage

[0242] In some aspects of the methods of treating a tendon injury’ in a subject in need thereof disclosed herein, the molecules comprise the structure:ATTORNEY DOCKET NO. 37759.0682P1Methods of Treating a Fracture

[0243] Disclosed are methods of treating a fracture in a subject in need thereofcomprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the fracture can be a bone stress injury such as a stress reaction or a stress fracture.

[0244] Disclosed herein are methods of treating a fracture in a subject in need thereof comprising administering to the subject molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-Pstructure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. Insome aspects of the methods of treating a fracture in a subject in need thereof disclosedherein, the bioactive moiety can be a variant of BMP -2 or a small molecule, such as, but not limited to, a piezol agonist, an IL- 1(3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0245] Disclosed herein are methods of treating a fracture in a subject in need thereof comprising administering to the subject molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-Pstructure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0246] Disclosed herein are methods of treating a fracture in a subject in need thereof comprising administering to the subject molecules comprising: i) a piezol agonist, an IL-1 (3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein thecleavable linker links the piezol agonist, IL- 1(3 antagonist, IRAK4 inhibitor, caspase- 1inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0247] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the molecule canATTORNEY DOCKET NO.37759.0682P1be administered locally. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0248] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0249] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP-2.

[0250] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159. Yaddlel. Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an IL- 10 antagonist, and the IL- 10 antagonist can be, but is not limited to, Di acerein (diacetylrhein). In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be a caspase- 1 inhibitor, and the caspase-1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide),ATTORNEY DOCKET NO.37759.0682P1Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be a saltinducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0251] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a nonnatural amino acid. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0252] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP -2. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-K3Plk. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of treating a fracture in a subj ect in need thereof disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of treatingATTORNEY DOCKET NO.37759.0682P1a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0253] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the variant of BMP-2 can be made using the methods described herein and in the Examples.

[0254] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0255] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGFβ-1, TGFβ-2, TGF-β3, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the tenogenic factor can be TGFβ-1 or TGFβ-2.

[0256] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11). bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0257] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of treating a fracture in a subj ect in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0258] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the MMPATTORNEY DOCKET NO.37759.0682P1cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0259] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697, and US 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0260] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC). 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP moiety is alendronate. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP moiety is zolendronate. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the BP moiety is pamidronate.iv. Spacer

[0261] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of treating a fracture in a subject in need thereof disclosedATTORNEY DOCKET NO.37759.0682P1herein, the spacer can be a peptide spacer. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70. 71. 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83. 84. 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95. 96. 97. 98. 99. 100, 101. 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166. 167, 168, 169, 170, 171. 172, 173. 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238. 239, 240, 241, 242, 243, 244, 245. 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0262] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0263] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0264] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0265] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the molecules comprise the structure:ATTORNEY DOCKET NO. 37759.0682P1AZIDE LINKERSpacer between BMP2 and Ctsk-peptideBISPHOSPHONATEConfers bone tissue affinityN-Gly-His-Pro-Gly-Gly-Pro-Gln-GlyPEPTIDE LINKERCtsk- substrate for cleavage

[0266] In some aspects of the methods of treating a fracture in a subject in need thereof disclosed herein, the molecules comprise the structure:4. Methods of Treating a Bone Stress Injury

[0267] Disclosed are methods of treating a bone stress injury in a subject in need thereof comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject.

[0268] Disclosed herein are methods of treating a bone stress injury in a subject in need thereof comprising administering to the subject molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0269] Disclosed herein are methods of treating a bone stress injury in a subject in need thereof comprising administering to the subject molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0270] Disclosed herein are methods of treating a bone stress injury’ in a subject in need thereof comprising administering to the subject molecules comprising: i) a piezol agonist, an IL- 1 [3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein theATTORNEY DOCKET NO.37759.0682P1cleavable linker links the piezol agonist, IL-1 [3 antagonist, IRAK4 inhibitor, caspase-1 inhibitor. NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0271] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of treating a bone stress injury' in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0272] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1. WISP-1 or CTGF.

[0273] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP-2.

[0274] In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yoda1, Yoda2 (KC289), KC159, Yaddle1, Jedi1 / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an IL-1β antagonist, and the IL-1β antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is notATTORNEY DOCKET NO.37759.0682P1limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of treating a tendon injury’ in a subject in need thereof disclosed herein, the bioactive moiety can be a caspase- 1 inhibitor, and the caspase-1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be. but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061. YKL-06-062, Dasatinib. Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of treating a tendon injury in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0275] In some aspects of the methods of treating a bone stress injury- in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0276] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of treating a bone stress injury- in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-K3Plk. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of treating a boneATTORNEY DOCKET NO.37759.0682P1stress injury' in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of treating a bone stress injury’ in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of treating a bone stress injury’ in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of treating a bone stress injury’ in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0277] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the variant of BMP-2 can be made using the methods described herein and in the Examples.

[0278] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor In some aspects, the osteogenic factor can be oxy 133 or oxysterol-133.

[0279] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the tenogenic factor can be an osteochondral factor, and the tenogenic factor can be, but is not limited to, TGF(3-1. TGF[ -2. TGF-(33, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the tenogenic factor can be TGF0-1 or TGFP-2.

[0280] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0281] In some aspects of the methods of treating a bone stress injury in a subject in needATTORNEY DOCKET NO.37759.0682P1thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0282] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0283] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697. and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912, US 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0284] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[1,2-c]pyridine-3-yl)-2-ATTORNEY DOCKET NO.37759.0682P1phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of treating a bone stress injury' in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0285] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7. 8, 9, 10, 11, 12, 13, 14, 15, 16, 17. 18. 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101. 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113. 114, 115, 116, 117, 118, 119. 120, 121, 122. 123, 124. 125, 126, 127, 128, 129. 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191. 192, 193, 194, 195, 196. 197, 198, 199, 200, 201, 202, 203. 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzy me accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0286] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link twoATTORNEY DOCKET NO.37759.0682P1different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0287] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the spacer can be, but is not limited, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0288] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0289] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityttyO" -' O' G G y H -Pro -Gly Giy -Pro Gi GtyPEPTIDE LINKERCtsk-substrate for cleavage

[0290] In some aspects of the methods of treating a bone stress injury in a subject in need thereof disclosed herein, the molecules comprise the structure:5. Methods of Enhancing Incorporation of Allograft Tissues During or After an Orthopedic Surgical Procedure

[0291] Disclosed are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject.

[0292] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof comprising administering molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) aATTORNEY DOCKET NO.37759.0682P1bisphosphonate moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, a piezol agonist, an IL-1 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK.1 inhibitor.

[0293] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof comprising administering molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0294] Disclosed herein are methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof comprising administering molecules comprising: i) a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL- 10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0295] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the orthopedic surgical procedure is an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

[0296] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the orthopedic surgical procedure is osteointegration of an implantable prosthesis. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the implantable prosthesis is an implant material, such as, but not limited to, metal, ceramic or plastic, that is implanted into bone or dental applications.

[0297] In some aspects of the methods of enhancing incorporation of allograft tissuesATTORNEY DOCKET NO.37759.0682P1during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the orthopedic surgical procedure is osteointegration of allograft tissue. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, allograft tissue is osteochondral allografts and / or bone allografts.

[0298] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule is administered just before the orthopedic surgical procedure. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule is administered during the orthopedic surgical procedure. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule is administered after the orthopedic surgical procedure.

[0299] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0300] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety is an ECM protein and the ECM protein can be,ATTORNEY DOCKET NO.37759.0682P1but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0301] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, BMP-1, BMP-2, TGF-01, TGF-02, TGF-03. BMP-3, BMP-4, BMP-5, BMP-6. BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP-2.

[0302] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a nonnatural amino acid.

[0303] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an IL-1β antagonist, and the IL-1β antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in need thereof disclosed herein, the bioactive moiety can be a caspase- 1 inhibitor, and the caspase- 1 inhibitor can be. but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein.ATTORNEY DOCKET NO.37759.0682P1the bioactive moiety can be aNLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312. MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0304] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-K3Plk. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of enhancingATTORNEY DOCKET NO.37759.0682P1incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-E83Azide. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0305] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP -2 can be made using the methods described herein and in the Examples.

[0306] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy 133 or oxysterol-133.

[0307] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF(3-1, TGF(3-2, TGF-(33. FGF, CTGF, BMP-12. BMP-13, BMP-14, CCN1 or WISP-1. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the tenogenic factor can be TGFP-1 or TGFP-2.

[0308] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein.ATTORNEY DOCKET NO.37759.0682P1growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0309] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0310] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0311] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697 and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0312] In some aspects of the methods of enhancing incorporation of allograft tissuesATTORNEY DOCKET NO.37759.0682P1during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1 -hydroxy -2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yljpropanoic acid (3-PEHPC). 2-hydroxy-3-(imidazo[1.2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0313] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59. 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84. 85. 86. 87. 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115,ATTORNEY DOCKET NO.37759.0682P1116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135. 136, 137, 138, 139, 140. 141, 142, 143, 144, 145, 146, 147. 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207. 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219. 220, 221, 222, 223, 224, 225. 226, 227, 228. 229, 230. 231, 232, 233, 234, 235. 236, 237. 238, 239, 240. 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0314] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0315] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0316] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0317] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecules comprise the structure:ATTORNEY DOCKET NO.37759.0682P1AZIDE LINKERSpacer between BMP2 and Ctsk-peptideBISPHOSPHONATEConfers bone tissue affinityN-; O "U' ■■■'■ O - ' ■ - N ■ Giy •• His -Pre -Giw G Pro-Gin -Giy''PEPTIDE LINKERCtsk- substrate for cleavage

[0318] In some aspects of the methods of enhancing incorporation of allograft tissues during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecules comprise the structure:6. Methods of Promoting Repair or Regeneration of Enthesis During or After an Orthopedic Surgical Procedure

[0319] Disclosed are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject.

[0320] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof comprising administering molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, a piezol agonist, an IL- 1(3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0321] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof comprising administering molecules comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein theATTORNEY DOCKET NO.37759.0682P1cleavable linker links the variant of BMP-2 to the BP moiety.

[0322] Disclosed herein are methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure comprising administering molecules comprising: i) a piezol agonist, an IL-1 antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL- 10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0323] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the orthopedic surgical procedure can be an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

[0324] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered just before the orthopedic surgical procedure. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered during the orthopedic surgical procedure. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered after the orthopedic surgical procedure.

[0325] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subj ect in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesisATTORNEY DOCKET NO.37759.0682P1ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0326] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be. but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0327] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be, but is not limited to, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the osteochondral factor can be parathyroid hormone. (PTH). parathyroid hormone-related protein (PTHrP), NELL- 1 or BMP-2.

[0328] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an IL-1β antagonist, and the IL-1β antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a caspase-ATTORNEY DOCKET NO.37759.0682P11 inhibitor, and the caspase- 1 inhibitor can be. but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01. YKL-05-099, YKL-06-061, YKL-06-062. Dasatinib, Bosutinib, MRT67307. or MRT 199665. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0329] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, atenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a nonnatural amino acid.

[0330] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedureATTORNEY DOCKET NO.37759.0682P1in a subject in need thereof disclosed herein, the variant of BMP -2 can be BMP2-K3Plk. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0331] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the variant of BMP -2 can be made using the methods described herein and in the Examples.

[0332] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor canATTORNEY DOCKET NO.37759.0682P1be oxy133 or oxysterol-133.

[0333] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF -1, TGF -2, TGF- 3, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the tenogenic factor can be TGFP-1 or TGF0-2.

[0334] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF 11 ), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0335] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0336] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ IDATTORNEY DOCKET NO.37759.0682P1NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0337] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697 and 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0338] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1 -hydroxy-2 -(pyridin-4-yl)ethane-l.l-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP). methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0339] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and theATTORNEY DOCKET NO.37759.0682P1bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84. 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101. 102, 103. 104, 105, 106, 107, 108. 109, 110, 111, 112, 113, 114, 115. 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171. 172, 173, 174, 175, 176. 177, 178, 179, 180, 181, 182, 183. 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243. 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0340] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0341] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.ATTORNEY DOCKET NO.37759.0682P1

[0342] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0343] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityPEPTIDE LINKER Ctsk-substrate for cleavage

[0344] In some aspects of the methods of promoting repair or regeneration of enthesis during or after an orthopedic surgical procedure in a subject in need thereof disclosed herein, the molecules comprise the structure:7. Methods of Treating Osteoporosis, Osteoarthritis and / or Cartilage Defects

[0345] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof comprising administering any one of the disclosed molecules, compositions or pharmaceutical compositions to the subject.

[0346] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof comprising administering a molecule comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as. but not limited to, aATTORNEY DOCKET NO.37759.0682P1piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0347] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof comprising administering a molecule comprising: i) a variant of BMP -2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0348] Disclosed herein are methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof comprising administering a molecule comprising: i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0349] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the site of osteoporosis, osteoarthritis and / or cartilage defects can be the knee, hip, ankle spine, wrist, shoulder, elbow, hands, fingers, feet and / or toes.

[0350] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecule can be administered systemically. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecule can be administered locally. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecule does not impair tissue healing or biomechanical strength. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, ectopic bone formation, heterotopic ossification, and enthesis ossification are decreased at the injury site of the subject.i. Bioactive Moiety

[0351] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subj ect in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aATTORNEY DOCKET NO.37759.0682P1NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be, but not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0352] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor is parathyroid hormone (PTH), parathyroid hormone related peptide, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP-2.

[0353] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yodal, Yoda2 (KC289), KC159, Yaddlel, Jedil / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an IL-1β antagonist, and the IL-1β antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a caspase- 1 inhibitor, and the caspase- 1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be. but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970. SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-ATTORNEY DOCKET NO.37759.0682P1099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0354] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0355] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subj ect in need thereof disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP-2. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a non-natural amino acid into BMP2. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP-2 comprises a propargyl-L-lysine incorporated into the amino acid sequence of BMP-2. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP -2 can be BMP2-K3Plk. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP -2 can be BMP2-A2C. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the bioactive moiety can be BMP2-A2C. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereofATTORNEY DOCKET NO.37759.0682P1disclosed herein the variant of BMP -2 can be BMP2-E83Plk. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the variant of BMP -2 can be BMP2 E94Plk. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0356] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the variant of BMP-2 can be made using the methods described herein and in the Examples.

[0357] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can be oxy133 or oxysterol-133.

[0358] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGF0-1, TGF0-2. TGF-03, FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP- 1. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein the tenogenic factor can be TGF0-1 or TGF0-2.

[0359] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptor-like kinase-l-Fc (ALK1-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0360] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) orATTORNEY DOCKET NO.37759.0682P1GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0361] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0362] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11.219.697, and US 11,840,549. all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and US 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0363] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yljpropanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-ATTORNEY DOCKET NO.37759.0682P1phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0364] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56. 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69. 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81. 82. 83. 84. 85. 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151. 152, 153, 154, 155, 156. 157, 158, 159, 160, 161, 162, 163. 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223. 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235. 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.ATTORNEY DOCKET NO. 37759.0682P1

[0365] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety ). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0366] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0367] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be an azide spacer. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0368] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityPEPTIDE LINKER Ctsk-substrate for cleavage

[0369] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecules comprise the structure:ATTORNEY DOCKET NO.37759.0682P18. Methods of Reducing Ectopic Bone Formation, Heterotopic Ossification, and Enthesis Ossification at an Injury site of the Subject

[0370] Disclosed are methods of reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site of a subject comprising administering to the subject an effective amount of any one of the molecules, compositions or pharmaceutical compositions disclosed herein to the subject.

[0371] Disclosed are methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject comprising administering a molecule comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate (BP) moiety, wherein the BP moiety has a P-C-P structure, and wherein the cleavable linker links the bioactive moiety to the BP moiety. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, or enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a variant of BMP-2 or a small molecule, such as, but not limited to, apiezol agonist, an IL- 1 [3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor.

[0372] Disclosed are methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject comprising administering a molecule comprising: i) a variant of BMP-2, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0373] Disclosed are methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject in comprising administering a molecule comprising: i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0374] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the injury site can be the knee, hip, ankle spine, wrist, shoulder, elbow, hands, fingers, feet and / or toes.

[0375] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, theATTORNEY DOCKET NO.37759.0682P1molecule can be administered systemically. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the molecule can be administered locally. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the molecule does not impair tissue healing or biomechanical strength.i. Bioactive Moiety

[0376] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor, an Extracellular Matrix (ECM) protein, a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor. In some aspects of methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an ECM protein, and the ECM protein can be, but is not limited to, a collagen, elastin, fibrin, fibronectin, gelatin, laminin, integrin, CCN1, WISP-1 or CTGF.

[0377] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an osteochondral factor, and the osteochondral factor can be parathyroid hormone (PTH), parathyroid hormone related peptide, NELL-1, TGF-β1, TGF-β2, TGF-β3, BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, or rhBMP. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the osteochondral factor can be NELL-1 or BMP -2.

[0378] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a piezol agonist, and the piezol agonist can be, but is not limited to, Yoda1, Yoda2 (KC289), KC159, Yaddle1, Jedi1 / 2, MCB-22-174, and CAS 3058199-62-2. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an IL-1β antagonist, and the IL-1β antagonist can be, but is not limited to, Diacerein (diacetylrhein). In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosedATTORNEY DOCKET NO.37759.0682P1herein, the bioactive moiety can be an IRAK4 inhibitor, and the IRAK4 inhibitor can be, but is not limited to, PF-06650833 (zimlovisertib). In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a caspase- 1 inhibitor, and the caspase-1 inhibitor can be, but is not limited to, VX-765 (belnacasan) or VX-740 (pralnacasan). In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a NLRP3 inflammasome inhibitor, and the NLRP3 inflammasome inhibitor can be, but is not limited to Glyburide (glibenclamide), Tranilast, MCC950, or Dapansutrile (OLT1177). In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a salt-inducible kinase inhibitor, and the salt inducible kinase inhibitor can be, but is not limited to GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, or MRT199665. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an Activin A receptor like type 1 (ALK1) inhibitor, and the ALK1 inhibitor can be, but is not limited to LDN214117.

[0379] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety has been genetic code expanded to comprise a non-natural amino acid. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, bioactive moiety can be an osteochondral factor, an osteogenic factor, a tenogenic factor or an Extracellular Matrix (ECM) protein and has been genetic code expanded to comprise a non-natural amino acid.

[0380] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an osteochondral factor and the osteochondral factor can be a variant of BMP -2. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2 that has been genetic code expanded to comprise a nonnatural amino acid into BMP2. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 comprises a propargyl-L-lysine incorporated into theATTORNEY DOCKET NO.37759.0682P1amino acid sequence of BMP-2. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2-K3Plk. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2-K3Plk. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2-E83stop. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2-E83stop. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2-A2C. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2-A2C. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2-E83Plk. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2-E83Plk. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2-E83Azide. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2-E83Azide. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be BMP2 E94Plk. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be BMP2 E94Plk.

[0381] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the variant of BMP-2 can be made using the methods described herein and in the Examples.

[0382] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be an osteogenic factor. In some aspects, the osteogenic factor can beATTORNEY DOCKET NO.37759.0682P1oxy 133 or oxysterol-133.

[0383] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be a tenogenic factor, and the tenogenic factor can be, but is not limited to, TGFβ-1, TGFβ-2, TGF-β3. FGF, CTGF, BMP-12, BMP-13, BMP-14, CCN1 or WISP-1. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the tenogenic factor can be TGFβ-1 or TGFβ-2.

[0384] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the bioactive moiety can be parathyroid hormone, parathyroid hormone-related protein, growth differentiation factor 11 (GDF11), bone morphogenic protein 11 (BMP-11), activin receptorlike kinase-l-Fc (ALKl-Fc), or activin receptor-like kinase-4-Fc (ALK4-Fc).ii. Cleavable Linker

[0385] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the cleavable linker can be a cathepsin-K-sensitive peptide linker. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the cathepsin-K-sensitive peptide linker can be GHPGGPQG (SEQ ID NO: 1).

[0386] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the cleavable linker can be an MMP cleavable linker. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury’ site of a subject disclosed herein, the MMP cleavable linkers can be PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18)ATTORNEY DOCKET NO.37759.0682P1or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0387] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the cleavable linker can be a pH sensitive linker. pH sensitive linkers can be those found in US Patent No. 8,063,209, 11,219,697, and US 11,840,549, all of which are hereby incorporated by reference in their entireties. pH sensitive linkers can also be found, for example, in US Patent No. 10,383,912 and US 10,688,193 both of which are hereby incorporated by reference in their entireties.iii. Bisphosphonate Moiety

[0388] In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the BP moiety does not fully or completely inhibit or only minimally inhibits osteoclast function. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the BP only minimally inhibits farnesyl pyrophosphate synthase (FPSS). In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the BP moiety can be, but is not limited to, 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP). methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury’ site of a subject disclosed herein, the BP moiety can be alendronate. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the BP moiety can be zolendronate. In some aspects of the methods of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject disclosed herein, the BP moiety can be pamidronate.iv. Spacer

[0389] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the compositions disclosed herein comprise a spacer between the cleavable linker and the bioactive moiety or the cleavable linker and the bisphosphonate moiety. In some aspects of the methods of treatingATTORNEY DOCKET NO.37759.0682P1osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be a peptide spacer. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer comprises less than 50 amino acids. The peptide spacer may comprise, comprise at least, or comprise at most 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31. 32. 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44. 45. 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56. 57. 58. 59. 60. 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115. 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133. 134, 135, 136, 137, 138. 139, 140, 141, 142, 143, 144, 145. 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205. 206, 207, 208, 209, 210. 211, 212, 213, 214, 215, 216, 217. 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, or 250 amino acids, or any derivable range therein. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer comprises more than 50 amino acids. Spacers can be used to reduce steric hindrance, promote enzyme accessibility and increase flexibility, improve the biological activity of the molecule or to improve the stability and bioavailability of the molecule.

[0390] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the peptide spacer can be a flexible molecule used to link two different molecules (e.g. the cleavable linker to the bioactive moiety or the cleavable linker to the bisphosphonate moiety ). Examples of spacers include, but are not limited to hydrophobic spacers, hydrophilic spacers, and PEGylated spacers.

[0391] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be, but is not limited to, a Beta-alanine, GABA, AEA, Ava, Ahx, PEG2 Spacer, PEG3 Spacer, PEG4 Spacer, or Ttds.

[0392] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the spacer can be an azideATTORNEY DOCKET NO. 37759.0682P1spacer. In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the azide spacer can be a PEG4-linker with a bioorthogonal azide group.

[0393] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecules comprise the structure:AZIDE LINKER BISPHOSPHONATESpacer between BMP2 and Ctsk-peptide Confers bone tissue affinityPEPTIDE LINKER Ctsk- substrate for cleavage

[0394] In some aspects of the methods of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof disclosed herein, the molecules comprise the structure:9. Dosing Regimens

[0395] Disclosed are dosing regimens comprising at least one treatment cycle of an effective amount of any of the disclosed molecules, compositions or pharmaceutical compositions.

[0396] Disclosed herein are dosing regimens in which the molecule, composition or pharmaceutical composition can be administered only once.

[0397] Disclosed herein are dosing regimens in which the molecule, composition or pharmaceutical composition can be administered multiple times over a period of time.

[0398] Disclosed herein are dosage regimens in which administration the molecule, composition or pharmaceutical composition can occur anywhere from day 0 (day of surgery) to 2 weeks after surgery.

[0399] Treatment cycles can include the administration of different dosages of molecules, compositions or pharmaceutical compositions as w ell as administration at different time points. The molecules, compositions or pharmaceutical compositions can be administered forATTORNEY DOCKET NO.37759.0682P1varying amounts of time for up to 6 months. The molecules, compositions or pharmaceutical compositions can be administered for varying amounts of time indefinitely. In some instances, the administration can occur for up to one, two, three, four, five or six months. For example, the molecule, composition or pharmaceutical composition can be administered once a week for 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, or 24 w eeks. In some aspects, the disclosed molecules, compositions or pharmaceutical compositions can be administered about every 3, about every' 6 or about every 12 months.

[0400] The length of time for each treatment cycle can vary depending on the amount of molecules, composition or pharmaceutical composition administered per dosage. A treatment cycle can include the administration of a molecule, composition or pharmaceutical composition once, twice or three times a week. In some aspects, the molecule, composition or pharmaceutical composition can be administered daily. In some aspects, the molecule, composition or pharmaceutical composition can be administered once every two w eeks or even once a month. In some instances, the molecule, composition or pharmaceutical composition can be administered every two weeks for 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, or 24 w eeks. For example, the treatment cycle can include administering a molecule, composition or pharmaceutical composition once a week for four weeks or once every' two weeks for up to six months. Thus, each treatment cycle includes an established length of time for administration as well as an established dosing schedule during that time frame.

[0401] In one aspect, more than molecule, composition or pharmaceutical composition can be administered during the treatment cycles. The more than one molecule, composition or pharmaceutical composition can be formulated together or in separate compositions. In some instances, one or more molecules, compositions or pharmaceutical compositions can be administered in combination with one or more other therapeutic agents, including, but not limited to antibodies, nanobodies, aptamers, liposomes, antioxidants, anti-inflammatory agents, senolytic agents.10. Dose

[0402] The dose or dosage of molecule, composition or pharmaceutical composition can vary depending on many factors, such as but not limited to, age, condition, sex and extent of the disease in the patient, route of administration, length of treatment cycle, or whether other drugs are included in the regimen, and can be determined by one of skill in the art.

[0403] Effective dosages can be determined empirically, and making such determinations is within the skill in the art. The dosage ranges for the administration of the compositions are those large enough to produce the desired effect in which the disease is treated. For example.ATTORNEY DOCKET NO.37759.0682P1the dosage can be an amount effective to provide therapeutic effects and provide or allow for sustained therapeutic effects even after the treatment (e.g. one or more of the disclosed molecules comprising: i) a bioactive moiety, ii) a cleavable linker, and iii) a bisphosphonate moiety, wherein the BP moiety has a P-C-P structure and wherein the cleavable linker links the bioactive moiety to the BP moiety) is withdrawn. The therapeutic effects can be, but are not limited to, an improvement in a tendon injury, the healing of a fracture or bone stress injury, a faster recovery from an orthopedic procedure, improved biomechanical properties, such as strength, or improved tissue organization of the repaired enthesis compared to a subject who did not receive the treatment. Other biomarkers used to measure therapeutic effects can be markers of tendon or bone repair. The therapeutic effects can be measured by imaging techniques, including MRI. intravascular ultrasound, ultrafast imaging CT scans, B-mode ultrasonography, virtual histology intravascular ultrasound, optical coherence tomography, or other known methods.

[0404] The dosage should not be so large as to cause adverse side effects, such as unwanted cross-reactions, anaphylactic reactions, and the like. The dosage can be adjusted by the individual physician in the event of any counter-indications. Dosage can vary, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products.

[0405] Suitable dosages include, but are not limited to amounts between 0.01 mg / kg and 20 mg / kg. For example, disclosed herein are methods involving administering one or more of the disclosed molecules, compositions or pharmaceutical compositions to a subject, wherein the molecule is administered in an amount of about 0.01 mg / kg to about 20 mg / kg. For example, the concentration of the molecule can be 0.01. 0.1, 1, 2, 3, 4, 5, 6. 7, 8, 9. 10. 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 mg / kg. Doses will depend on the identity of the bioactive moiety. Doses of BP-BMP-2, for example, may be about 10 mg / kg.

[0406] Suitable dosages include, but are not limited to, amounts between 0.01 pg and 100 pg. For example, disclosed herein are methods involving administering one or more of the disclosed molecules, compositions or pharmaceutical compositions to a subject, wherein the molecule is administered in an amount of about 0.01 pg to about 100 pg. For example, the concentration of the molecule can be 0.01 pg, 0.05 pg, 0.1 pg, 0.2 pg, 0.3 pg, 0.4 pg, 0.5 pg, 0.6 pg, 0.7 pg, 0.8 pg, 0.9 pg, 1 pg, 2 pg, 3 pg, 4 pg, 5 pg, 6 pg, 7 pg, 8 pg, 9 pg, 10 pg, 11 pg. 12 pg, 13 pg, 14 pg. 15 pg, 16 pg, 17 pg, 18 pg, 19 pg. 20 pg, 21 pg, 22 pg, 23 pg, 24 pg, 25 pg, 26 pg, 27 pg, 28 pg, 29 pg, 30 pg, 35 pg, 40 pg, 45 pg, 50 pg, 55 pg, 60ATTORNEY DOCKET NO.37759.0682P1μg, 65 μg, 70 μg, 75 μg, 80 μg, 85 μg, 90 μg, 95 μg, or 100 μg. Doses will depend on the identity- of the bioactive moiety. Doses of BP-BMP-2, for example, may be about 5 pg.

[0407] The molecule, composition or pharmaceutical composition dose can be administered systemically or locally. The molecule, composition or pharmaceutical composition dose can be administered as a bolus injection or as an infusion over one or more hours.11. Delivery

[0408] In the methods described herein, administration or delivery of the molecules, compositions or pharmaceutical compositions can be via a variety of mechanisms. As defined above, disclosed herein are methods of treating, dosing regimens and methods of using those dosing regimens to treat. The dosing regimens and methods include compositions containing any one or more of the molecules described herein that can also include a carrier such as a pharmaceutically acceptable carrier. For example, disclosed are pharmaceutical compositions, comprising the molecules and compositions disclosed herein, and a pharmaceutically acceptable carrier.

[0409] The disclosed molecules, compositions or pharmaceutical compositions can be in solution or in suspension (for example, incorporated into microparticles, liposomes, or cells).

[0410] Any suitable route of administration can be used for the disclosed molecules, compositions and pharmaceutical compositions. Suitable routes of administration can, for example, include topical, enteral, local, systemic, or parenteral. For example, administration can be epicutaneous, inhalational, enema, conjunctival, eye drops, ear drops, alveolar, nasal, intranasal, enteral, oral, intraoral, transoral, intestinal, rectal, intrarectal, transrectal, injection, infusion, intravenous, intraarterial, intramuscular, intracerebral, intraventricular, intracerebroventricular, intracardiac, subcutaneous, intraosseous, intradermal, intrathecal, intraperitoneal, intravesical, intracavemosal, intramedullar, intraocular, intracranial, transdermal, transmucosal, transnasal, inhalational, intracistemal, epidural, peridural, intravitreal, etc. The disclosed compositions can be used in and with any other therapy.

[0411] In another embodiment, one or more components of the solution can be provided as a "concentrate", e.g., in a storage container (e.g., in a premeasured volume) ready for dilution, or in a soluble capsule ready for addition to a volume of water.

[0412] The foregoing formulations and administration methods are intended to be illustrative and not limiting. It will be appreciated that, using the teaching provided herein, other suitable formulations and modes of administration can be readily devised.ATTORNEY DOCKET NO.37759.0682P112. Combination Therapy

[0413] In one aspect of the disclosed methods, molecules, compositions or pharmaceutical compositions can be administered alone or in combination with one or more additional therapeutic agents. The additional therapeutic agents are selected based on the disease or symptom to be treated. A description of the various classes of suitable pharmacological agents and drugs may be found in Goodman and Gilman, The Pharmacological Basis of Therapeutics, (11th Ed., McGraw-Hill Publishing Co.) (2005). For example, pharmaceutical compositions containing molecules can be administered in combination with one or more known therapeutic agents for treating atherosclerosis.

[0414] Examples of therapeutic agents that treat tendon injuries include, but are not limited to, anti-inflammatory agents, analgesic agents, anti-rheumatologic agents, and immune modulating agents, biophysical agents (e.g. shock wave therapy, ultrasound, magnetic fields, joint and tissue passive motion devices), dry needling, thermal therapy (e.g. ice, heat) and therapeutic exercises (e.g. physical therapy, eccentric strengthening).

[0415] The molecules, compositions or pharmaceutical compositions can be administered in conjunction with or followed by any of the disclosed additional therapeutics. In some aspects, the BP-BMP -2 molecule can be administered in combination with parathyroid hormone (PTH). In some aspects, the BP-BMP -2 molecule can be administered in combination with bisphosphonate parathyroid hormone (BP-PTH).The combination therapies can include administering the molecule, composition or pharmaceutical composition and an additional therapeutic agent during the treatment cycle of a dosing regimen.D. Embodiments

[0416] Embodiment 1. A molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0417] Embodiment 2. The molecule of embodiment 1. wherein the molecule further comprises a spacer.

[0418] Embodiment 3. The molecule of embodiment 2, wherein the linker is an azideATTORNEY DOCKET NO.37759.0682P1spacer.

[0419] Embodiment 4. The molecule of embodiment 3, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0420] Embodiment 5. The molecule of any one of embodiments 1 to 4, wherein the variant of BMP-2 is BMP2-K3Plk, BMP2-E83stop, BMP2-A2C, BMP2-E83Plk, BMP2-E83Azide, or BMP2 E94Plk.

[0421] Embodiment 6. The molecule of any one of embodiments 1 to 5, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0422] Embodiment 7. The molecule of embodiment 6, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0423] Embodiment 8. The molecule of embodiment 7, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0424] Embodiment 9. The molecule of any one of embodiments 1 to 5, wherein the cleavable linker is an MMP cleavable linker.

[0425] Embodiment 10. The molecule of embodiment 9, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0426] Embodiment 11. The molecule of any one of embodiments 1 to 5, wherein the cleavable linker is a pH sensitive linker.

[0427] Embodiment 12. The molecule of any one of embodiments 1 to 11, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0428] Embodiment 13. The molecule of any one of embodiments 1 to 12, wherein the BP moiety is 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[1,2-c]pyridine-3-yl)-2-ATTORNEY DOCKET NO.37759.0682P1phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0429] Embodiment 14. The molecule of embodiment 13, wherein the BP moiety is pamidronate.

[0430] Embodiment 15. A composition comprising the molecule of any one of embodiments 1 to 14.

[0431] Embodiment 16. A pharmaceutical composition comprising the molecule of any one of embodiments 1 to 14 or the composition of embodiment 15.

[0432] Embodiment 17. A method of targeting a molecule to a site of injury to a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP -2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0433] Embodiment 18. The method of embodiment 17, wherein the site of injury is enthesis tissue.

[0434] Embodiment 19. The method of embodiment 17 or embodiment 18, wherein the site of injury is at a rotator cuff, a distal biceps tendon, a pectoralis major, a patellar tendon, a quadriceps tendon, or a triceps tendon.

[0435] Embodiment 20. The method of embodiment 17 or embodiment 18, wherein the site of injury is also a site of surgery and the surgery is an anterior cruciate ligament (ACL) reconstruction, a posterior cruciate ligament (PCL) reconstruction, a lateral collateral ligament (LCL) or medial collateral ligament (MCL) repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, or an osteochondral allograft transplantation.

[0436] Embodiment 21. The method of any one of embodiments 17 to 20, wherein the site of injury is at the site of osteointegration of implantable prosthesis or osteointegration of allograft tissue.

[0437] Embodiment 22. The method of any one of embodiments 17 to 21, wherein the molecule is administered systemically.

[0438] Embodiment 23. The method of any one of embodiments 17 to 21, wherein the molecule is administered locally.ATTORNEY DOCKET NO.37759.0682P1

[0439] Embodiment 24. The method of any one of embodiments 17 to 23, wherein the molecule further comprises a spacer.

[0440] Embodiment 25. The method of embodiment 24, wherein the linker is an azide spacer.

[0441] Embodiment 26. The method of embodiment 25, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0442] Embodiment 27. The method of any one of embodiments 17 to 26, wherein the variant of BMP-2 is BMP2-K3Plk, BMP2-E83stop, BMP2-A2C, BMP2-E83Plk, BMP2-E83Azide, or BMP2 E94Plk.

[0443] Embodiment 28. The method of any one of embodiments 17 to 27, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0444] Embodiment 29. The method of embodiment 28, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0445] Embodiment 30. The method of embodiment 29, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0446] Embodiment 31. The method of any one of embodiments 17 to 27, wherein the cleavable linker is a MMP cleavable linker.

[0447] Embodiment 32. The method of embodiment 31, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0448] Embodiment 33. The method of any one of embodiments 17 to 27, wherein the cleavable linker is a pH sensitive linker.

[0449] Embodiment 34. The method of any one of embodiments 17 to 33, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0450] Embodiment 35. The method of any one of embodiments 17 to 34, wherein the BP moiety is 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-ATTORNEY DOCKET NO.37759.0682P1(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[1,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0451] Embodiment 36. The method embodiment 35, wherein the BP moiety is pamidronate.

[0452] Embodiment 37. A method of treating a tendon injury, fracture and / or bone stress injury and / or promoting soft tissue to bone healing to a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP -2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0453] Embodiment 38. The method of embodiment 37, wherein the tendon injury is a rotator cuff, a distal biceps tendon, pectoralis major, patellar tendon, quadriceps tendon, and / or triceps tendon.

[0454] Embodiment 39. A method of enhancing incorporation of allograft tissues or promoting repair of enthesis during or after an orthopedic surgical procedure to a subject in need thereof comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0455] Embodiment 40. The method of embodiment 39, wherein the orthopedic surgical procedure is an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

[0456] Embodiment 41. The method of embodiment 39 or embodiment 40, wherein the molecule is administered just before the orthopedic surgical procedure.ATTORNEY DOCKET NO.37759.0682P1

[0457] Embodiment 42. The method of embodiment 39 or embodiment 40, wherein the molecule is administered during the orthopedic surgical procedure.

[0458] Embodiment 43. The method of embodiment 39 or embodiment 40, wherein the molecule is administered after the orthopedic surgical procedure.

[0459] Embodiment 44. A method of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0460] Embodiment 45. A method of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

[0461] Embodiment 46. The method of any one of embodiments 37 to 45, wherein the molecule is administered systemically.

[0462] Embodiment 47. The method of any one of embodiments 37 to 45, wherein the molecule is administered locally at the site of the tendon injury or surgical procedure.

[0463] Embodiment 48. The method of any one of embodiments 37 to 47, wherein the molecule further comprises a spacer.

[0464] Embodiment 49. The method of embodiment 48, wherein the linker is an azide spacer.

[0465] Embodiment 50. The method of embodiment 49, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0466] Embodiment 51. The method of any one of embodiments 37 to 50, wherein the variant of BMP-2 is BMP2-K3Plk, BMP2-E83stop, BMP2-A2C, BMP2-E83Plk. BMP2-E83Azide, or BMP2 E94Plk.ATTORNEY DOCKET NO.37759.0682P1

[0467] Embodiment 52. The method of any one of embodiments 37 to 51, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0468] Embodiment 53. The method of embodiment 52, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0469] Embodiment 54. The method of embodiment 53, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0470] Embodiment 55. The method of any one of embodiments 37 to 51, wherein the cleavable linker is a MMP cleavable linker.

[0471] Embodiment 56. The method of embodiment 55, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0472] Embodiment 57. The method of any one of embodiments 37 to 51, wherein the cleavable linker is a pH sensitive linker.

[0473] Embodiment 58. The method of any one of embodiments 37 to 57, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0474] Embodiment 59. The method of any one of embodiments 37 to 58, wherein the BP moiety is 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[1,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0475] Embodiment 60. The method of embodiment 59, wherein the BP moiety is pamidronate.

[0476] Embodiment 61. The method of any one of embodiments 17 to 60, wherein the molecule is administered at a dosage of 0.01 pg to about 100 pg or 0.01 mg / kg to about 20 mg / kg.ATTORNEY DOCKET NO.37759.0682P1

[0477] Embodiment 62. The method of any one of embodiments 17 to 61, wherein the molecule does not impair tissue healing or biomechanical strength.

[0478] Embodiment 63. The method of any one of embodiments 17 to 62, wherein the molecule decreases ectopic bone formation, heterotopic ossification, and enthesis ossification in the subject.

[0479] Embodiment 64. A molecule comprising:i) a piezol agonist, an IL- 1(3 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL- 1 (3 antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0480] Embodiment 65. The molecule of embodiment 64, wherein the molecule further comprises a spacer.

[0481] Embodiment 66. The molecule of embodiment 65, wherein the linker is an azide spacer.

[0482] Embodiment 67. The molecule of embodiment 66, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0483] Embodiment 68. The molecule of any one of embodiments 64 to 67, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedi 1 / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glybunde, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN- 112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

[0484] Embodiment 69. The molecule of any one of embodiments 64 to 68, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0485] Embodiment 70. The molecule of embodiment 69, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0486] Embodiment 71. The molecule of embodiment 69, wherein the cathepsin-K-ATTORNEY DOCKET NO.37759.0682P1sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0487] Embodiment 72. The molecule of any one of embodiments 64 to 68, wherein the cleavable linker is an MMP cleavable linker.

[0488] Embodiment 73. The molecule of embodiment 72, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0489] Embodiment 74. The molecule of any one of embodiments 64 to 68, wherein the cleavable linker is a pH sensitive linker.

[0490] Embodiment 75. The molecule of any one of embodiments 64 to 74, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0491] Embodiment 76. The molecule of any one of embodiments 64 to 74, wherein the BP moiety is 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0492] Embodiment 77. The molecule of embodiment 76, wherein the BP moiety is pamidronate.

[0493] Embodiment 78. A composition comprising the molecule of any one of embodiments 64 to 77.

[0494] Embodiment 79. A method of targeting a molecule to a site of injury to a subject in need thereof, comprising administering a molecule comprising:i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;ATTORNEY DOCKET NO.37759.0682P1wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0495] Embodiment 80. The method of embodiment 79, wherein the site of injury is enthesis tissue.

[0496] Embodiment 81. The method of embodiment 79 or embodiment 80, wherein the site of injury is at a rotator cuff, a distal biceps tendon, a pectoralis major, a patellar tendon, a quadriceps tendon, or a triceps tendon.

[0497] Embodiment 82. The method of embodiment 79 or embodiment 80, wherein the site of injury is also a site of surgery and the surgery is an anterior cruciate ligament (ACL) reconstruction, a posterior cruciate ligament (PCL) reconstruction, a lateral collateral ligament (LCL) or medial collateral ligament (MCL) repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, or an osteochondral allograft transplantation.

[0498] Embodiment 83. The method of any one of embodiments 79 to 82, wherein the site of injury is at the site of osteointegration of implantable prosthesis or osteointegration of allograft tissue.

[0499] Embodiment 84. The method of any one of embodiments 79 to 83, wherein the molecule is administered systemically.

[0500] Embodiment 85. The method of any one of embodiments 79 to 83, wherein the molecule is administered locally.

[0501] Embodiment 86. The method of any one of embodiments 79 to 85, wherein the molecule further comprises a spacer.

[0502] Embodiment 87. The method of embodiment 86, wherein the linker is an azide spacer.

[0503] Embodiment 88. The method of embodiment 87, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0504] Embodiment 89. The method of any one of embodiments 78 to 88, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedi 1 / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glybunde, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.ATTORNEY DOCKET NO.37759.0682P1

[0505] Embodiment 90. The method of any one of embodiments 79 to 89, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0506] Embodiment 91. The method of embodiment 90, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0507] Embodiment 92. The method of embodiment 91, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0508] Embodiment 93. The method of any one of embodiments 79 to 89, wherein the cleavable linker is a MMP cleavable linker.

[0509] Embodiment 94. The method of embodiment 93, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0510] Embodiment 95. The method of any one of embodiments 79 to 89, wherein the cleavable linker is a pH sensitive linker.

[0511] Embodiment 96. The method of any one of embodiments 79 to 96, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0512] Embodiment 97. The method of any one of embodiments 79 to 96, wherein the BP moiety is 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[1,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0513] Embodiment 98. The method embodiment 97, wherein the BP moiety is pamidronate.

[0514] Embodiment 99. A method of treating a tendon injury, fracture and / or bone stress injury and / or promoting soft tissue to bone healing to a subject in need thereof, comprising administering a molecule comprising:ATTORNEY DOCKET NO.37759.0682P1i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0515] Embodiment 100. The method of embodiment 99, wherein the tendon injury is a rotator cuff, a distal biceps tendon, pectoralis major, patellar tendon, quadriceps tendon, and / or triceps tendon.

[0516] Embodiment 101. A method of enhancing incorporation of allograft tissues or promoting repair of enthesis during or after an orthopedic surgical procedure to a subject in need thereof comprising administering a molecule comprising:i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety’.

[0517] Embodiment 102. The method of embodiment 101, wherein the orthopedic surgical procedure is an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

[0518] Embodiment 103. The method of embodiment 101 or embodiment 102, wherein the molecule is administered just before the orthopedic surgical procedure.

[0519] Embodiment 104. The method of embodiment 101 or embodiment 102, wherein the molecule is administered during the orthopedic surgical procedure.

[0520] Embodiment 105. The method of embodiment 101 or embodiment 102, wherein the molecule is administered after the orthopedic surgical procedure.ATTORNEY DOCKET NO.37759.0682P1

[0521] Embodiment 106. A method of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof, comprising administering a molecule comprising:i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

[0522] Embodiment 107. The method of any one of embodiments 99 to 106, wherein the molecule is administered systemically.

[0523] Embodiment 108. The method of any one of embodiments 99 to 106, wherein the molecule is administered locally at the site of the tendon injury or surgical procedure.

[0524] Embodiment 109. The method of any one of embodiments 99 to 108, wherein the molecule further comprises a spacer.

[0525] Embodiment 110. The method of embodiment 109, wherein the linker is an azide spacer.

[0526] Embodiment 111. The method of embodiment 110, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0527] Embodiment 112. The method of any one of embodiments 99 to 111, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel. Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833. VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

[0528] Embodiment 113. The method of any one of embodiments 99 to 112, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0529] Embodiment 114. The method of embodiment 113, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.ATTORNEY DOCKET NO.37759.0682P1

[0530] Embodiment 115. The method of embodiment 114, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

[0531] Embodiment 116. The method of any one of embodiments 99 to 108, wherein the cleavable linker is a MMP cleavable linker.

[0532] Embodiment 117. The method of embodiment 116, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0533] Embodiment 118. The method of any one of embodiments 99 to 108, wherein the cleavable linker is a pH sensitive linker.

[0534] Embodiment 119. The method of any one of embodiments 99 to 118, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0535] Embodiment 120. The method of any one of embodiments 99 to 119, wherein the BP moiety is 2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy-2-(pyridin-4-yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP). methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3-(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine-3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

[0536] Embodiment 121. The method of embodiment 120, wherein the BP moiety is pamidronate.

[0537] Embodiment 122. The method of any one of embodiments 79 to 121, wherein the molecule is administered at a dosage of 0.01 pg and to about 100 pg or 0.01 mg / kg to about 20 mg / kg.

[0538] Embodiment 123. The method of any one of embodiments 79 to 122, wherein the molecule does not impair tissue healing or biomechanical strength.

[0539] Embodiment 124. The method of any one of embodiments 79 to 123, wherein the molecule decreases ectopic bone formation, heterotopic ossification, and enthesis ossification in the subject.

[0540] Embodiment 125. A method of reducing ectopic bone formation, heterotopicATTORNEY DOCKET NO.37759.0682P1ossification, and enthesis ossification at an injury site of a subject comprising:i) a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL- 10 antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety’.

[0541] Embodiment 126. The method of embodiment 125, wherein the molecule is administered systemically.

[0542] Embodiment 127. The method of embodiment 125 or embodiment 126, wherein the molecule is administered locally at the site of the tendon injury or surgical procedure.

[0543] Embodiment 128. The method of any one of embodiments 125 to 127, wherein the molecule further comprises a spacer.

[0544] Embodiment 129. The method of embodiment 128, wherein the linker is an azide spacer.

[0545] Embodiment 130. The method of embodiment 129, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

[0546] Embodiment 131. The method of any one of embodiments 125 to 130, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061. YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

[0547] Embodiment 132. The method of any one of embodiments 125 to 131, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

[0548] Embodiment 133. The method of embodiment 132, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

[0549] Embodiment 134. The method of embodiment 133, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).ATTORNEY DOCKET NO.37759.0682P1

[0550] Embodiment 135. The method of any one of embodiments 125 to 134, wherein the cleavable linker is a MMP cleavable linker.

[0551] Embodiment 136. The method of embodiment 135, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD-GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

[0552] Embodiment 137. The method of any one of embodiments 125 to 136, wherein the cleavable linker is a pH sensitive linker.

[0553] Embodiment 138. The method of any one of embodiments 125 to 137, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

[0554] Embodiment 139. The method of any one of embodiments 125 to 138, wherein the BP moiety is 2-(pyridin-4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1-hydroxy- 2-(pyridin-4-yl)ethane-1,1-diyl...

Claims

1. CLAIMSWe claim:

1. A molecule comprising:i) a variant of BMP-2:ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

2. The molecule of claim 1, wherein the molecule further comprises a spacer.

3. The molecule of claim 2, wherein the linker is an azide spacer.

4. The molecule of claim 3, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

5. The molecule of any one of claims 1 to 4. wherein the variant of BMP -2 is BMP2- K3Plk, BMP2-E83stop, BMP2-A2C, BMP2-E83Plk, BMP2-E83Azide, or BMP2 E94Plk.

6. The molecule of any one of claims 1 to 5, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

7. The molecule of claim 6, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

8. The molecule of claim 7. wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

9. The molecule of any one of claims 1 to 5, wherein the cleavable linker is an MMP cleavable linker.

10. The molecule of claim 9, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).ATTORNEY DOCKET NO.37759.0682P111. The molecule of any one of claims 1 to 5, wherein the cleavable linker is a pH sensitive linker.

12. The molecule of any one of claims 1 to 11, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

13. The molecule of any one of claims 1 to 12, wherein the BP moiety is 2-(pyridin-4- yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

14. The molecule of claim 13, wherein the BP moiety is pamidronate.

15. A composition comprising the molecule of any one of claims 1 to 14.

16. A pharmaceutical composition comprising the molecule of any one of claims 1 to 14 or the composition of claim 15.

17. A method of targeting a molecule to a site of injury to a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the variant of BMP-2to the BP moiety.

18. The method of claim 17, wherein the site of injury is enthesis tissue.

19. The method of claim 17 or claim 18, wherein the site of injury is at a rotator cuff, a distal biceps tendon, a pectoralis major, a patellar tendon, a quadriceps tendon, or a triceps tendon.

20. The method of claim 17 or claim 18, wherein the site of injury is also a site of surgery and the surgery is an anterior cruciate ligament (ACL) reconstruction, a posterior cruciate ligament (PCL) reconstruction, a lateral collateral ligament (LCL) or medial collateral ligament (MCL) repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, or an osteochondral allograft transplantation.

21. The method of any one of claims 17 to 20, wherein the site of injury is at the site of osteointegration of implantable prosthesis or osteointegration of allograft tissue.ATTORNEY DOCKET NO. 37759.0682P122. The method of any one of claims 17 to 21, wherein the molecule is administered systemically.

23. The method of any one of claims 17 to 21. wherein the molecule is administered locally.

24. The method of any one of claims 17 to 23, wherein the molecule further comprises a spacer.

25. The method of claim 24, wherein the linker is an azide spacer.

26. The method of claim 25, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

27. The method of any one of claims 17 to 26, wherein the variant of BMP-2 is BMP2-K3Plk, BMP2-E83stop, BMP2-A2C. BMP2-E83Plk, BMP2-E83Azide, or BMP2 E94Plk.

28. The method of any one of claims 17 to 27, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

29. The method of claim 28, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

30. The method of claim 29, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

31. The method of any one of claims 17 to 27. wherein the cleavable linker is a MMP cleavable linker.

32. The method of claim 31, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

33. The method of any one of claims 17 to 27, wherein the cleavable linker is a pH sensitive linker.

34. The method of any one of claims 17 to 33, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

35. The method of any one of claims 17 to 34, wherein the BP moiety is 2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-PyrEBP). 1 -hydroxy -2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP). 2-hydroxy-2-phosphono-3-ATTORNEY DOCKET NO.37759.0682P1(pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

36. The method claim 35, wherein the BP moiety is pamidronate.

37. A method of treating a tendon injury, fracture and / or bone stress injury and / or promoting soft tissue to bone healing to a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety:wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the variant of BMP-2to the BP moiety.

38. The method of claim 37, wherein the tendon injury’ is a rotator cuff, a distal biceps tendon, pectoralis major, patellar tendon, quadriceps tendon, and / or triceps tendon.

39. A method of enhancing incorporation of allograft tissues or promoting repair of enthesis during or after an orthopedic surgical procedure to a subject in need thereof comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the variant of BMP-2to the BP moiety.

40. The method of claim 39, wherein the orthopedic surgical procedure is an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

41. The method of claim 39 or claim 40. wherein the molecule is administered just before the orthopedic surgical procedure.

42. The method of claim 39 or claim 40, wherein the molecule is administered during the orthopedic surgical procedure.

43. The method of claim 39 or claim 40, wherein the molecule is administered after the orthopedic surgical procedure.ATTORNEY DOCKET NO. 37759.0682P144. A method of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof, comprising administering a molecule comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety:wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the variant of BMP-2to the BP moiety.

45. A method of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject comprising:i) a variant of BMP-2;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety7has a PCP structure, andwherein the cleavable linker links the variant of BMP-2 to the BP moiety.

46. The method of any one of claims 37 to 45. wherein the molecule is administered systemically.

47. The method of any one of claims 37 to 45, wherein the molecule is administered locally at the site of the tendon injury' or surgical procedure.

48. The method of any one of claims 37 to 47, wherein the molecule further comprises a spacer.

49. The method of claim 48, wherein the linker is an azide spacer.

50. The method of claim 49, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

51. The method of any one of claims 37 to 50, wherein the variant of BMP-2 is BMP2-K3Plk, BMP2-E83stop, BMP2-A2C, BMP2-E83Plk, BMP2-E83Azide, or BMP2 E94Plk.

52. The method of any one of claims 37 to 51, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

53. The method of claim 52, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.ATTORNEY DOCKET NO.37759.0682P154. The method of claim 53, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

55. The method of any one of claims 37 to 51. wherein the cleavable linker is a MMP cleavable linker.

56. The method of claim 55, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

57. The method of any one of claims 37 to 51, wherein the cleavable linker is a pH sensitive linker.

58. The method of any one of claims 37 to 57, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

59. The method of any one of claims 37 to 58, wherein the BP moiety is 2-(pyridin-4- yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP). 1 -hydroxy -2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

60. The method of claim 59, wherein the BP moiety is pamidronate.

61. The method of any one of claims 17 to 60, wherein the molecule is administered at a dosage of 0.01 pg to about 100 pg or 0.01 mg / kg to about 20 mg / kg.

62. The method of any one of claims 17 to 61, wherein the molecule does not impair tissue healing or biomechanical strength.

63. The method of any one of claims 17 to 62, wherein the molecule decreases ectopic bone formation, heterotopic ossification, and enthesis ossification in the subject.

64. A molecule comprising:i) a piezol agonist, an IL- 1 P antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andhi) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andATTORNEY DOCKET NO.37759.0682P1wherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase-1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

65. The molecule of claim 64, wherein the molecule further comprises a spacer.

66. The molecule of claim 65, wherein the linker is an azide spacer.

67. The molecule of claim 66, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

68. The molecule of any one of claims 64 to 67, wherein the piezol agonist, an IL-1 P antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF- 06650833, VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

69. The molecule of any one of claims 64 to 68, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

70. The molecule of claim 69, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

71. The molecule of claim 69, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

72. The molecule of any one of claims 64 to 68, wherein the cleavable linker is an MMP cleavable linker.

73. The molecule of claim 72, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

74. The molecule of any one of claims 64 to 68, wherein the cleavable linker is a pH sensitive linker.

75. The molecule of any one of claims 64 to 74, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

76. The molecule of any one of claims 64 to 74, wherein the BP moiety is 2-(pyridin- 4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-(pyridin-4-ATTORNEY DOCKET NO.37759.0682P1yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

77. The molecule of claim 76, wherein the BP moiety is pamidronate.

78. A composition comprising the molecule of any one of claims 64 to 77.

79. A method of targeting a molecule to a site of injury to a subject in need thereof, comprising administering a molecule comprising:i) a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a P-C-P structure, andwherein the cleavable linker links the piezol agonist, IL-10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

80. The method of claim 79, wherein the site of injury is enthesis tissue.

81. The method of claim 79 or claim 80, wherein the site of injury’ is at a rotator cuff, a distal biceps tendon, a pectoralis major, a patellar tendon, a quadriceps tendon, or a triceps tendon.

82. The method of claim 79 or claim 80, wherein the site of injury’ is also a site of surgery and the surgery is an anterior cruciate ligament (ACL) reconstruction, a posterior cruciate ligament (PCL) reconstruction, a lateral collateral ligament (LCL) or medial collateral ligament (MCL) repair / reconstruction. a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, or an osteochondral allograft transplantation.

83. The method of any one of claims 79 to 82, wherein the site of injury' is at the site of osteointegration of implantable prosthesis or osteointegration of allograft tissue.

84. The method of any one of claims 79 to 83. wherein the molecule is administered systemically.

85. The method of any one of claims 79 to 83, wherein the molecule is administered locally.

86. The method of any one of claims 79 to 85, wherein the molecule further comprises a spacer.ATTORNEY DOCKET NO.37759.0682P187. The method of claim 86, wherein the linker is an azide spacer.

88. The method of claim 87, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

89. The method of any one of claims 78 to 88, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

90. The method of any one of claims 79 to 89. wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

91. The method of claim 90, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

92. The method of claim 91, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

93. The method of any one of claims 79 to 89, wherein the cleavable linker is a MMP cleavable linker.

94. The method of claim 93, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

95. The method of any one of claims 79 to 89, wherein the cleavable linker is a pH sensitive linker.

96. The method of any one of claims 79 to 96, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

97. The method of any one of claims 79 to 96, wherein the BP moiety is 2-(pyridin-4- yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), 1 -hydroxy -2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

98. The method claim 97, wherein the BP moiety is pamidronate.ATTORNEY DOCKET NO.37759.0682P199. A method of treating a tendon injury, fracture and / or bone stress injury and / or promoting soft tissue to bone healing to a subject in need thereof, comprising administering a molecule comprising:i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-1β antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

100. The method of claim 99, wherein the tendon injury is a rotator cuff, a distal biceps tendon, pectoralis major, patellar tendon, quadriceps tendon, and / or triceps tendon.

101. A method of enhancing incorporation of allograft tissues or promoting repair of enthesis during or after an orthopedic surgical procedure to a subject in need thereof comprising administering a molecule comprising:i) a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, aNLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL- 10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

102. The method of claim 101, wherein the orthopedic surgical procedure is an osteotomy, an ACL reconstruction, a PCL reconstruction, an LCL or MCL repair / reconstruction, a medial patellofemoral ligament reconstruction, a meniscus root repair, a meniscus allograft transplantation, and / or osteochondral allograft transplantation.

103. The method of claim 101 or claim 102, wherein the molecule is administered just before the orthopedic surgical procedure.

104. The method of claim 101 or claim 102, wherein the molecule is administered during the orthopedic surgical procedure.ATTORNEY DOCKET NO.37759.0682P1105. The method of claim 101 or claim 102, wherein the molecule is administered after the orthopedic surgical procedure.

106. A method of treating osteoporosis, osteoarthritis and / or cartilage defects in a subject in need thereof, comprising administering a molecule comprising:i) a piezol agonist, an IL- 10 antagonist, an IRAK4 inhibitor, a caspase- 1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety7has a PCP structure, andwherein the cleavable linker links the piezol agonist, IL-10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety7.

107. The method of any one of claims 99 to 106, wherein the molecule is administered systemically.

108. The method of any one of claims 99 to 106, wherein the molecule is administered locally at the site of the tendon injury7or surgical procedure.

109. The method of any one of claims 99 to 108, wherein the molecule further comprises a spacer.

110. The method of claim 109, wherein the linker is an azide spacer.

111. The method of claim 110, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

112. The method of any one of claims 99 to 111, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutimb, MRT67307, MRT199665, or LDN214117.

113. The method of any one of claims 99 to 112, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

114. The method of claim 113, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

115. The method of claim 114, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).ATTORNEY DOCKET NO.37759.0682P1116. The method of any one of claims 99 to 108, wherein the cleavable linker is a MMP cleavable linker.

117. The method of claim 116, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

118. The method of any one of claims 99 to 108, wherein the cleavable linker is a pH sensitive linker.

119. The method of any one of claims 99 to 118, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

120. The method of any one of claims 99 to 119, wherein the BP moiety is 2-(pyridin- 4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP), l-hydroxy-2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP), 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

121. The method of claim 120, wherein the BP moiety is pamidronate.

122. The method of any one of claims 79 to 121, wherein the molecule is administered at a dosage of 0.01 pg and to about 100 pg or 0.01 mg / kg to about 20 mg / kg.

123. The method of any one of claims 79 to 122, wherein the molecule does not impair tissue healing or biomechanical strength.

124. The method of any one of claims 79 to 123, wherein the molecule decreases ectopic bone formation, heterotopic ossification, and enthesis ossification in the subject.

125. A method of reducing ectopic bone formation, heterotopic ossification, and enthesis ossification at an injury site of a subject comprising:i) a piezol agonist, an IL-1β antagonist, an IRAK4 inhibitor, a caspase-1 inhibitor, a NLRP3 inflammasome inhibitor, a salt-inducible kinase inhibitor, or an ALK1 inhibitor;ii) a cleavable linker; andiii) a bisphosphonate (BP) moiety;wherein the BP moiety has a PCP structure, andATTORNEY DOCKET NO.37759.0682P1wherein the cleavable linker links the piezol agonist, IL- 10 antagonist, IRAK4 inhibitor, caspase- 1 inhibitor, NLRP3 inflammasome inhibitor, salt-inducible kinase inhibitor, or ALK1 inhibitor to the BP moiety.

126. The method of claim 125, wherein the molecule is administered systemically.

127. The method of claim 125 or claim 126, wherein the molecule is administered locally at the site of the tendon injury or surgical procedure.

128. The method of any one of claims 125 to 127, wherein the molecule further comprises a spacer.

129. The method of claim 128, wherein the linker is an azide spacer.

130. The method of claim 129, wherein the azide spacer is a PEG4-linker with a bioorthogonal azide group.

131. The method of any one of claims 125 to 130, wherein the agonist, antagonist, or inhibitor is Yodal, Yoda2, KC159, Yaddlel, Jedil / 2, MCB-22-174, CAS 3058199-62-2, Diacerein, PF-06650833, VX-765, VX-740, Glyburide, Tranilast, MCC950, Dapansutrile, GLPG3970, SK-124, ARN-3236, GLPG3312, MRIA9, OMX-0407, Pterosin B, KIN-112, HG-9-91-01, YKL-05-099, YKL-06-061, YKL-06-062, Dasatinib, Bosutinib, MRT67307, MRT199665, or LDN214117.

132. The method of any one of claims 125 to 131, wherein the cleavable linker is a cathepsin-K-sensitive peptide linker.

133. The method of claim 132, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1) or GGGMGPSGPWGGK (SEQ ID NO: 2) or HPGGPQ (SEQ ID NO: 21) or KPRGSKQ (SEQ ID NO: 22) or KKPGSKQ (SEQ ID NO: 23) or Z-Gly-Pro-Arg-AMC.

134. The method of claim 133, wherein the cathepsin-K-sensitive peptide linker is GHPGGPQG (SEQ ID NO: 1).

135. The method of any one of claims 125 to 134, wherein the cleavable linker is a MMP cleavable linker.

136. The method of claim 135, wherein the MMP cleavable linker is PLGLAG (SEQ ID NO: 3) or Ac-GCRD-GPQGIWGQ-DRCG (SEQ ID NO: 4) or Ac-GCRD- GPQGIAGQ-DRCG (SEQ ID NO: 5) or SGESPAYYTA (SEQ ID NO: 9) or GAPFALRLV (SEQ ID NO: 10) or GGYAELRMGG (SEQ ID NO: 11) or GPLGLWAR (SEQ ID NO: 12) or GPLGMRGL (SEQ ID NO: 13) or IPESLRAG (SEQ ID NO: 14) or PVGLIG (SEQ ID NO: 15) or PLGLAG (SEQ ID NO: 16) or PLGVR (SEQ ID NO: 17) or VPMSMRGG (SEQ ID NO: 18) or GPQGIWGQ (SEQ ID NO: 19) or GPQGIAGQ (SEQ ID NO: 20).

137. The method of any one of claims 125 to 136, wherein the cleavable linker is a pH sensitive linker.ATTORNEY DOCKET NO.37759.0682P1138. The method of any one of claims 125 to 137, wherein the BP moiety does not inhibit osteoclast function and / or only minimally inhibits farnesyl pyrophosphate synthase (FPSS).

139. The method of any one of claims 125 to 138, wherein the BP moiety is 2-(pyridin- 4-yl)ethane-l,l-diylbisphosphonic acid (p-PyrEBP). l-hydroxy-2-(pyridin-4- yl)ethane-1,1-diylbisphosphonic acid (p-RIS), methylene bisphosphonate (MBP), methylene hydroxyl bisphosphonate (MHDP). 2-hydroxy-2-phosphono-3- (pyridine-3-yl)propanoic acid (3-PEHPC), 2-hydroxy-3-(imidazo[l,2-c]pyridine- 3-yl)-2-phosphonopropanoic acid (3-IP-EHPC), alendronate, zolendronate, or pamidronate.

140. The method of claim 139, wherein the BP moiety is pamidronate.

141. The method of any one of claims 125 to 140, wherein the molecule is administered at a dosage of 0.01 pg and to about 100 pg or 0.01 mg / kg to about 20 mg / kg.

142. The method of any one of claims 125 to 141, wherein the molecule does not impair tissue healing or biomechanical strength.

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