Trans-cyclooctene bioorthogonal agents and uses in cancer and immunotherapy

Abstract

To provide compositions and methods that may be used for bioorthogonal delivery to a targeted location in a subject.SOLUTION: Trans-cyclooctene conjugates of therapeutic agents may be used for bioorthogonal delivery to a targeted location in a subject. The compositions and methods have applications in the treatment of various diseases or conditions including cancer, tumor growths, and bacterial infections.SELECTED DRAWING: Figure 1A

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 871,051, filed July 5, 2019, U.S. Provisional Patent Application No. 62 / 971,196, filed February 6, 2020, and U.S. Provisional Patent Application No. 62 / 981,401, filed February 25, 2020, each of which is incorporated by reference in its entirety herein.

[0002] The present disclosure provides trans-cyclooctene derivatives and uses for bioorthogonal delivery to a subject for cancer and / or immunotherapy. [Background technology]

[0003] Immunotherapies are being clinically tested to enhance the immune system's response to tumor growth and cancer dissemination. Immunotherapeutic strategies utilize immune cells and include monoclonal antibodies against tumor antigens, immune checkpoint inhibitors, vaccination, adoptive immune cell therapy (e.g., CAR-T cells), and cytokine administration.

[0004] TLR agonists play a fundamental role in the activation of innate and adaptive immune responses. In mouse models, treatment with TLR agonists has been shown to suppress tumor growth and, in some cases, destroy established tumors when used in combination with other therapeutics, such as chemotherapeutic drugs, mAbs, and various tumor antigen vaccines in the form of proteins, peptides, or plasmid DNA. TLR agonists activate professional antigen-presenting cells (APCs), i.e., dendritic cells (DCs). TLRs induce favorable antitumor effects by inducing inflammatory cytokine expression and cytotoxic T lymphocyte (CTL) responses. As adjuvants, TLR agonists can initiate strong immune responses to support cancer radiotherapy and biochemotherapy. Recently, it has been demonstrated that TLR binding to various T cell subsets increases their responses, which represents a novel and promising strategy to enhance the efficacy of cancer immunotherapy.

[0005] The central role of STING in regulating anti-cancer immune responses was illustrated by the observation that spontaneous and radiation-induced adaptive immunity was reduced in the absence of STING, indicating the potential of targeting STING for cancer immunotherapy.

[0006] Bioorthogonal conjugation reactions, or click reactions, are selective and orthogonal (non-interacting) functionalities found in biological systems that are useful for a variety of applications in chemistry, chemical biology, molecular diagnostics, and medicine, where they can be used to facilitate the selective manipulation of molecules, cells, particles, and surfaces, as well as the tagging and tracking of biomolecules in vitro and in vivo. These reactions include the Staudinger ligation, the azide-cyclooctyne cycloaddition, and the inverse electron-demand Diels-Alder reaction.

[0007] WO 2017 / 044983 describes the antitumor effect of a trans-cyclooctene conjugate of doxorubicin, obtained by release of doxorubicin at the tumor site via a bioorthogonal reaction with a tetrazine-functionalized alginate implanted at the tumor site. Summary of the Invention [Means for solving the problem]

[0008] The present disclosure provides trans-cyclooctene derivatives for delivering payload molecules to a subject using bioorthogonal chemistry. The present disclosure also provides methods of producing the compositions, as well as methods of using the compositions.

[0009] In one aspect, the present invention provides a compound of formula (I), or a pharma- ceutically acceptable salt thereof: [ka] (In the formula, R 1a Each occurrence of represents hydrogen, C 1~4 Alkyl, and C 1~4 independently selected from the group consisting of haloalkyl; R 1b Each occurrence of represents hydrogen, C 1~4 Alkyl, C 1~4 Haloalkyl, C(O)OH, C(O)OC 1~4 Alkyl, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )CHR 1e C(O)OC 1~4 Alkyl, C(O)N(R 1c )-C 1~6 Alkylene-COH and C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 independently selected from the group consisting of alkyl; R 1c Each occurrence is independently hydrogen or C 1~4 is alkyl; R 1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; D, at each occurrence, is independently a payload selected from the group consisting of a toll-like receptor (TLR) agonist and a stimulator of interferon genes (STING) agonist; L 1 is, independently at each occurrence, a linker; m is independently at each occurrence 1, 2, or 3; p, at each occurrence, is independently 0, 1, or 2. to provide.

[0010] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0011] In another aspect, the present invention provides a method of treating or preventing a disease state or disorder or enhancing or eliciting an immune response, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I), or a pharma- ceutically acceptable salt or composition thereof; and a therapeutic support composition, the therapeutic support composition comprising a biocompatible support and a compound of formula [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S) is selected from the group consisting of O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R'', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; t is 0, 1, 3, or 4. The tetrazine-containing group is

[0012] In another aspect, the present invention provides a pharmaceutical combination comprising a compound of formula (I), or a pharma- ceutically acceptable salt or composition thereof, for use in the treatment or prophylaxis of a disease or disorder, such as cancer, infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or inducing an immune response; and a therapeutic support composition, said therapeutic support composition comprising a biocompatible support, as defined herein, and a compound of formula [ka] The tetrazine-containing group is

[0013] In another aspect, the present invention provides the use of a combination comprising a compound of formula (I), or a pharma- ceutically acceptable salt or composition thereof; and a therapeutic support composition, in the manufacture of a medicament for the treatment or prevention of a condition or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, graft rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or eliciting an immune response, said therapeutic support composition comprising a biocompatible support, as defined herein, and a compound of formula [ka] The tetrazine-containing group is

[0014] Aspects of the present disclosure include a method of delivering an effective amount of a payload to a target site in a subject, the method comprising administering to the subject a therapeutic support composition as defined herein.

[0015] Aspects of the present disclosure also include kits comprising a compound of formula (I), a therapeutic support composition as defined herein, and optionally a compound of formula (IB) as defined herein.

[0016] Another aspect of the present invention provides a method of treating cancer or enhancing or inducing an immune response, comprising administering to a subject in need thereof: a) a therapeutically effective amount of a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof. [ka] (In the formula, R 1A Each time it occurs, C 1~4 Alkyl, C 1~4 Haloalkyl and C 1~4 independently selected from the group consisting of alkoxy; R 1B Each time it occurs, G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1~6 Alkylene-COH, -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-COH)-C 2~4 Alkylene) n -N(C 1~4 Alkylene -COH)2, -N(R 1c )CHR 1e C(O)OC 1~6 Alkyl, -N(R 1c )-C 1~6 Alkylene-C(O)OC 1~6 Alkyl, and -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-C(O)OC 1~6 Alkyl)-C 2~4 Alkylene) n -N(C 1~4 Alkylene-C(O)OC 1~6alkyl)2; R 1c and R 1d Each occurrence is independently hydrogen or C 1~4 is alkyl; R 1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; R 1f Each occurrence independently represents hydrogen, C 1~6 Alkyl or -C 1~4 alkylene-COH; D 1 is, each occurrence independently, an anticancer drug payload; L 1 is, independently at each occurrence, a linker; L 2 Each occurrence independently represents -C(O)- and C 1~3 alkylene; G 1 is independently at each occurrence an optionally substituted heterocyclyl; m is 1, 2, or 3; n is independently at each occurrence 0, 1, 2, or 3; p, at each occurrence, is independently 0, 1, or 2; b) a support and a compound of formula [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2 is selected from the group consisting of NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S) R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' are independently selected from hydrogen, aryl, and alkyl at each occurrence; R''' is independently selected from aryl and alkyl at each occurrence; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; and t is 0, 1, 3, or 4. a therapeutic support composition comprising a tetrazine-containing group of the formula (wherein the tetrazine-containing group is linked or directly bonded to the support); c) a therapeutically effective amount of one or more immunomodulatory agents, or pharma- ceutically acceptable salts thereof; The method includes administering

[0017] In another aspect, the present invention provides a pharmaceutical combination of a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; a therapeutic support composition; and an immunomodulatory agent for use in the treatment or prevention of a disease or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or inducing an immune response.

[0018] In another aspect, the present invention provides the use of a combination of a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; a therapeutic support composition; and an immunomodulatory agent in the manufacture of a medicament for the treatment or prevention of a condition or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or inducing an immune response.

[0019] Another aspect of the invention provides a kit comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt or composition thereof; b) an immunomodulatory agent, or a pharma- ceutically acceptable salt or composition thereof; and c) instructions for use.

[0020] Another aspect of the invention provides a kit comprising: a) a therapeutic support composition; b) an immunomodulatory agent, or a pharma- ceutically acceptable salt or composition thereof; and c) instructions for use.

[0021] Another aspect of the present invention provides a pharmaceutical composition comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof; b) an immunomodulatory agent, or a pharma- ceutically acceptable salt thereof; and c) a pharma- ceutically acceptable carrier.

[0022] Another aspect of the present invention provides a pharmaceutical composition comprising: a) a therapeutic support composition; b) an immunomodulatory agent, or a pharma- ceutically acceptable salt thereof; and c) a pharma- ceutically acceptable carrier.

[0023] Another aspect of the present disclosure includes a method of delivering an effective amount of a payload to a target site in a subject, the method comprising administering to the subject a therapeutic support composition as defined herein.

[0024] Another aspect of the present invention is a method for producing a composition comprising the steps of: a) a therapeutically effective amount of a compound of formula (II-A), or a pharma- ceutically acceptable salt thereof. [ka] (In the formula, R 1A Each time it occurs, C 1~4 Alkyl, C 1~4 Haloalkyl and C 1~4 independently selected from the group consisting of alkoxy; R 1B Each time it occurs, G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1~6 Alkylene-COH, -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-COH)-C 2~4 Alkylene) n -N(C 1~4 Alkylene -COH)2, -N(R 1c )CHR 1e C(O)OC 1~6 Alkyl, -N(R 1c )-C 1~6 Alkylene-C(O)OC 1~6 Alkyl, and -N(R1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-C(O)OC 1~6 Alkyl)-C 2~4 Alkylene) n -N(C 1~4 Alkylene-C(O)OC 1~6 alkyl)2; R 1c and R 1d Each occurrence is independently hydrogen or C 1~4 is alkyl; R 1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; R 1f Each occurrence independently represents hydrogen, C 1~6 Alkyl or -C 1~4 alkylene-COH; D 1 is, each occurrence independently, an anticancer drug payload; L 1 is, independently at each occurrence, a linker; L 2 Each occurrence of -C(O)- and C 1~3 independently selected from the group consisting of alkylene; G 1 is independently at each occurrence an optionally substituted heterocyclyl; m is 1, 2, or 3; n is independently at each occurrence 0, 1, 2, or 3; p, at each occurrence, is independently 0, 1, or 2. to a subject in need thereof; and b) a support and a compound of formula [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'' , NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; t is 0, 1, 3, or 4. locally administering to the subject a therapeutic support composition comprising a tetrazine-containing group of the formula: and providing a method for treating cancer comprising administering to said patient a In this case, the subject has a second tumor, and administration of a) and administration of b) inhibits the growth of the second tumor in the patient.

[0025] In another aspect, the present invention provides a method of enhancing or inducing an immune response against a second tumor in a subject, the method comprising the steps of: a) administering to the subject a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof; and b) locally administering to a first tumor in the subject a therapeutic support composition, wherein the compound of Formula (II-A) or (III-A) and the therapeutic support composition are as defined herein, and wherein administering a) and administering b) enhances or induces an immune response against the second tumor.

[0026] In another aspect, the present invention provides a method of inhibiting tumor metastasis in a subject at risk of tumor metastasis, the method comprising the steps of: a) administering to the subject a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof; and b) locally administering to a first tumor of the subject a therapeutic support composition, wherein the compound of Formula (II-A) or (III-A) and the therapeutic support composition are as defined herein. do.

[0027] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, for use in a method for inhibiting the growth of a second tumor in a subject, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0028] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, for use in a method of enhancing or inducing an immune response against a second tumor in a subject, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0029] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, for use in a method of inhibiting tumor metastasis in a subject at risk of tumor metastasis, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0030] In another aspect, the present invention provides the use of a combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof; and b) a therapeutic support composition, in the manufacture of a medicament for inhibiting the growth of a second tumor, wherein the therapeutic support composition is administered locally to a first tumor in a subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof, is administered to the subject.

[0031] In another aspect, the present invention provides the use of a combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof; and b) a therapeutic support composition, in the manufacture of a medicament for enhancing or inducing an immune response against a second tumor, wherein the therapeutic support composition is administered locally to a first tumor in a subject, and a compound of formula (IA), or a pharma- ceutically acceptable salt, or composition thereof, is administered to the subject.

[0032] In another aspect, the present invention provides the use of a combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof; and b) a therapeutic support composition, in the manufacture of a medicament for inhibiting tumor metastasis in a subject at risk of tumor metastasis, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof, is administered to the subject. [Brief description of the drawings]

[0033] [Figure 1A] Figure 1 shows the body weight effect of mice injected with MC38 colorectal tumor cells into the right flank and then treated at the tumor injection site with modified sodium hyaluronate biomaterial in combination with systemic treatment with saline (G1), prodrug 1 (G2), or prodrug 1 and a TLR9a agonist (G3) as described in Examples B1 and C1. Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 1B] Figure 1 shows the body weight effect of mice injected with MC38 colorectal tumor cells in both the right and left flanks, followed by treatment with modified sodium hyaluronate biomaterial at the tumor injection site in the right flank in combination with systemic treatment with saline (G4), doxorubicin (G5), doxorubicin and TLR9a agonist (G6), prodrug 1 and TLR9a agonist (G7), or prodrug 1 (G8), as described in Example C1. Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 2A] Figure 1 shows the effect on tumor size in mice injected with MC38 colorectal tumor cells into the right flank followed by treatment of the tumor injection site with modified sodium hyaluronate biomaterial in combination with systemic treatment with saline (G1), prodrug 1 (G2), or prodrug 1 and a TLR9a agonist (G3) as described in Examples B1 and C1. Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 2B] As described in Example C1, the effect on right flank tumor size of mice injected with MC38 colorectal tumor cells in both the right and left flanks, followed by treatment of the right flank tumor injection site with modified sodium hyaluronate biomaterial in combination with systemic treatment with saline (G4), doxorubicin (G5), doxorubicin and TLR9a agonist (G6), prodrug 1 and TLR9a agonist (G7), or prodrug 1 (G8). Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 3A]Figure 1 shows the effect on tumor size in mice injected with MC38 colorectal tumor cells into the right flank followed by treatment of the tumor injection site with modified sodium hyaluronate biomaterial in combination with systemic treatment with saline (G1), prodrug 1 (G2), or prodrug 1 and a TLR9a agonist (G3) as described in Examples B1 and C1. Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 3B] As described in Example C1, the effect on right flank tumor size of mice injected with MC38 colorectal tumor cells in both the right and left flanks, followed by treatment of the right flank tumor injection site with modified sodium hyaluronate biomaterial in combination with systemic treatment with saline (G4), doxorubicin (G5), doxorubicin and TLR9a agonist (G6), prodrug 1 and TLR9a agonist (G7), or prodrug 1 (G8). Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 4] As described in Example C1, the effect on left flank tumor size of mice treated with modified sodium hyaluronate biomaterial at the tumor injection site on the right flank in combination with systemic treatment with saline (G4), doxorubicin (G5), doxorubicin and TLR9a agonist (G6), prodrug 1 and TLR9a agonist (G7), or prodrug 1 (G8) after injection of MC38 colorectal tumor cells into both the right and left flanks. Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Diagram 5] As described in Example C1, the effect on left flank tumor size of mice injected with MC38 colorectal tumor cells in both the right and left flanks, followed by treatment of the right flank tumor injection site with modified sodium hyaluronate biomaterial in combination with saline (G4), doxorubicin (G5), doxorubicin and TLR9a agonist (G6), prodrug 1 and TLR9a agonist (G7), or prodrug 1 (G8). Data points represent group mean body weight. Error bars represent standard error of the mean (SEM). [Figure 6A]The treatment schedules for treatment groups G4, G5, and G8 are shown. [Figure 6B] 1 shows a comparison of the effect on volumetric increase of biomaterial-injected right flank tumors observed in treatment groups G4, G5, and G8. The shaded area represents the Biomaterial 1 / Prodrug 1 treatment period. [Figure 6C] 1 shows a comparison of the effect on non-biomaterial injected left flank tumor volume increase observed in treatment groups G4, G5, and G8. The shaded area represents the Biomaterial 1 / Prodrug 1 treatment period. [Figure 6D] Kaplan-Meier survival curves for mice in treatment groups G4, G5, and G8 are shown. [Figure 7A] Tumor growth curves for individual mice in treatment group G8 are shown. [Figure 7B] Tumor growth curves for individual mice in treatment group G5 are shown. [Figure 7C] Tumor growth curves for individual mice in treatment group G4 are shown. [Figure 8] FIG. 13 shows tumor infiltrating immune cell profile in biomaterial injected right flank tumors 2 weeks after treatment for treatment group G8. [Figure 9] FIG. 1 shows tumor infiltrating immune cell profile 2 weeks after treatment in left flank tumors not injected with biomaterial for treatment group G8. [Figure 10A] As described in Example C1, MC38 colorectal tumor cells were injected into the right flank (day 0), and the tumor injection site was treated with the modified sodium hyaluronate biomaterial in combination with systemic treatment with prodrug 1 (G2), followed by the effect on tumor size in one mouse that received a second injection of MC38 colorectal tumor cells into the left flank on day 70 (indicated by the arrow in FIG. 10A). [Figure 10B] A comparison of the effects of five naive mice injected with MC38 colorectal tumor cells on the same day with the treatment group in FIG. 10A is shown. [Figure 11]Kaplan-Meier survival curves for mice in treatment groups G4, G6, and G7 are shown. ***Statistical significance of survival times was determined by the log-rank (Mantel-Cox) test. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] 1.Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present specification, including definitions, shall control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

[0035] As used herein, "comprising," "including," "having," "having," "can," "containing," and variations thereof are intended to be open-ended transitional phrases that do not exclude the possibility of additional acts or structures. The singular forms "a," "an," and "the" include plural referents unless the context clearly requires otherwise. The present disclosure also contemplates other embodiments that "comprise," "consist of," and "consist essentially of" the embodiments or elements presented herein, whether or not explicitly stated.

[0036] The modifier "about" used in connection with a quantity is inclusive of the recited value and has the meaning required by the context (e.g., it includes at least the degree of error associated with measurement of the particular quantity). The modifier "about" should also be considered as disclosing a range defined by the absolute values ​​of the two endpoints. For example, the phrase "about 2 to about 4" also discloses a range of "2 to 4". The term "about" may refer to plus or minus 10% of the indicated number. For example, "about 10%" may indicate a range of 9% to 11%, and "about 1" may mean 0.9 to 1.1. Other meanings of "about" may be apparent from the context, such as rounding, for example, "about 1" may also mean 0.5 to 1.4.

[0037] The conjunction "or" includes any combination of one or more of the listed elements that the conjunction connects. For example, the phrase "an apparatus comprising A or B" can refer to an apparatus that comprises A and B is absent, an apparatus that comprises B and A is absent, or an apparatus that comprises both A and B. The phrases "at least one of A, B, . . . and N" and "at least one of A, B, . . . and N" can be used to refer to any combination of one or more of the listed elements that the conjunction connects. The phrase "at least one of A, B, ...N, or combinations thereof" is broadly defined to mean one or more elements selected from the group including A, B, ..., and N, or in other words, any combination of one or more of the elements A, B, ..., or N, including any one element by itself or in combination with one or more other elements that may also include other unlisted elements in combination.

[0038] Definitions of specific functional groups and chemical terms are described in more detail below. For purposes of this disclosure, chemical elements are defined as defined by the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th Edition, thOrganic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March's Advanced Organic Chemistry, 5th Ed., 1999, inside cover, and specific functional groups are generally defined as described therein. Further, general principles of organic chemistry and specific functional groups and reactivity are described in the following: Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3 rd Edition, Cambridge University Press, Cambridge, 1987; each of which is incorporated herein by reference in its entirety.

[0039] The term "alkoxy," as used herein, refers to an alkyl group (as defined herein) appended to the parent molecular moiety through an oxygen atom. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, 2-propoxy, butoxy, and tert-butoxy.

[0040] As used herein, the term "alkyl" refers to a linear or branched saturated hydrocarbon chain containing 1 to 30 carbon atoms. The term "lower alkyl" or "C1-C6-alkyl" refers to a linear or branched hydrocarbon chain containing 1 to 6 carbon atoms. The term "C1-C3-alkyl" refers to a linear or branched hydrocarbon chain containing 1 to 3 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, n-heptyl, n-octyl, n-nonyl, and n-decyl.

[0041] As used herein, the term "alkenyl" refers to a hydrocarbon chain containing 2 to 30 carbon atoms and having at least one carbon-carbon double bond. The alkenyl group may be substituted or unsubstituted. For example, the alkenyl group may be substituted with an aryl group, such as, for example, phenyl.

[0042] As used herein, the term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbonyl group containing 2 to 30 carbon atoms, e.g., 2 to 20 or 2 to 10 carbon atoms, and having at least one site of triple bond unsaturation. The term "alkyne" also includes non-aromatic cycloalkyl groups of 5 to 20 carbon atoms, e.g., 5 to 10 carbon atoms, having single or multiple rings and with at least one triple bond. Examples of such alkynyl groups include, but are not limited to, acetylenyl (-C≡CH), and propargyl (-CH2C≡CH), as well as cycloalkynyl moieties, such as, but not limited to, aryl ... , substituted or unsubstituted cyclooctyne moieties.

[0043] The term "alkoxyalkyl," as used herein, refers to an alkoxy group, as defined herein, appended to the parent molecular moiety through an alkyl group.

[0044] As used herein, the term "alkylene" refers to a divalent group derived from a straight or branched chain hydrocarbon of 1 to 30 carbon atoms, e.g., 2 to 10 carbon atoms. Representative examples of alkylene include, but are not limited to, -CH2CH2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, and -CH2CH2CH2CH2CH2-.

[0045] The term "amino acid" refers to both natural and unnatural amino acids. It also includes protected natural and unnatural amino acids.

[0046] As used herein, the term "aryl" refers to a phenyl group, or a bicyclic aryl or tricyclic aryl fused ring system. An example of a bicyclic fused ring system includes a phenyl group attached to a parent molecular moiety and fused with one phenyl group. An example of a tricyclic fused ring system includes a phenyl group attached to a parent molecular moiety and fused with two other phenyl groups. Representative examples of bicyclic aryl include, but are not limited to, naphthyl. Representative examples of tricyclic aryl include, but are not limited to, anthracenyl. Monocyclic, bicyclic, and tricyclic aryl are connected to the parent molecular moiety through any carbon atom contained within their ring and may be unsubstituted or substituted.

[0047] As used herein, the term "azide" refers to the functional group -N3.

[0048] As used herein, the term "cycloalkyl" refers to a carbocyclic ring system containing 3 to 10 carbon atoms, 0 heteroatoms, and 0 double bonds. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl. "Cycloalkyl" also encompasses a carbocyclic ring system in which a cycloalkyl is appended to a parent molecular moiety and is fused to an aryl group, as defined herein, a heteroaryl group, as defined herein, or a heterocycle, as defined herein.

[0049] As used herein, the term "cycloalkenyl" means a non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and preferably having from 5 to 10 carbon atoms per ring. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, or cycloheptenyl.

[0050] As used herein, the term "cyclooctene" refers to a substituted or unsubstituted non-aromatic cyclic alkyl group of 8 carbon atoms having a single ring with a double bond. Examples of such cyclooctene groups include, but are not limited to, substituted or unsubstituted trans-cyclooctene (TCO).

[0051] As used herein, the term "fluoroalkyl" means an alkyl group (as defined herein) in which one, two, three, four, five, six, seven, or eight hydrogen atoms are replaced by fluorine. Representative examples of fluoroalkyl include, but are not limited to, 2-fluoroethyl, 2,2,2-trifluoroethyl, trifluoromethyl, difluoromethyl, pentafluoroethyl, and trifluoropropyl, such as 3,3,3-trifluoropropyl.

[0052] The term "alkoxyfluoroalkyl," as used herein, refers to an alkoxy group (as defined herein) appended to the parent molecular moiety through a fluoroalkyl group (as defined herein).

[0053] The term "fluoroalkoxy," as used herein, means at least one fluoroalkyl group (as defined herein) is appended to the parent molecular moiety through an oxygen atom. Representative examples of fluoroalkoxy include, but are not limited to, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy.

[0054] As used herein, the term "halogen" or "halo" means Cl, Br, I, or F.

[0055] As used herein, the term "haloalkyl" means an alkyl group (as defined herein) in which 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms are replaced by halogen.

[0056] As used herein, the term "haloalkoxy" means at least one haloalkyl group (as defined herein) is appended to the parent molecular moiety through an oxygen atom.

[0057] As used herein, the term "heteroalkyl" refers to an alkyl group (as defined herein) in which one or more carbon atoms are replaced by a heteroatom selected from S, Si, O, P, and N. The heteroatom may be optionally oxidized. Representative examples of heteroalkyl include, but are not limited to, alkyl ethers, secondary and tertiary alkyl amines, and alkyl sulfides.

[0058] As used herein, the term "heteroaryl" refers to an aromatic monocyclic ring or aromatic bicyclic ring system or aromatic tricyclic ring system. An aromatic monocyclic ring is a 5- or 6-membered ring containing at least one heteroatom independently selected from the group consisting of N, O, and S (e.g., 1, 2, 3, or 4 heteroatoms independently selected from O, S, and N). A 5-membered aromatic monocyclic ring has two double bonds, and a 6-membered aromatic monocyclic ring has three double bonds. Examples of bicyclic heteroaryl groups include a monocyclic heteroaryl ring attached to a parent molecular moiety and fused with a monocyclic cycloalkyl group, as defined herein, a monocyclic aryl group, as defined herein, a monocyclic heteroaryl group, as defined herein, or a monocyclic heterocycle, as defined herein. Examples of tricyclic heteroaryl groups include a monocyclic heteroaryl ring appended to a parent molecular moiety and fused to two of a monocyclic cycloalkyl group, a monocyclic aryl group, a monocyclic heteroaryl group, or a monocyclic heterocycle, as defined herein. Representative examples of monocyclic heteroaryls include, but are not limited to, pyridinyl (including pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl), pyrimidinyl, pyrazinyl, thienyl, furyl, thiazolyl, thiadiazolyl, isoxazolyl, pyrazolyl, and 2-oxo-1,2-dihydropyridinyl. Representative examples of bicyclic heteroaryls include, but are not limited to, chloromethyl, benzothienyl, benzodioxolyl, benzotriazolyl, quinolinyl, thienopyrrolyl, thienothienyl, imidazothiazolyl, benzothiazolyl, benzofuranyl, indolyl, quinolinyl, imidazopyridine, benzoxadiazolyl, and benzopyrazolyl. Representative examples of tricyclic heteroaryls include, but are not limited to, dibenzofuranyl and dibenzothienyl. Monocyclic, bicyclic, and tricyclic heteroaryls are connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the ring and can be unsubstituted or substituted. Either is acceptable.

[0059] As used herein, the term "heterocycle" refers to monocyclic heterocycles, bicyclic heterocycles, and tricyclic heterocycles. Monocyclic heterocycles are 3-, 4-, 5-, 6-, 7-, or 8-membered rings containing at least one heteroatom independently selected from the group consisting of O, N, and S. 3- or 4-membered rings contain zero or one double bond and one heteroatom selected from the group consisting of O, N, and S. 5-membered rings contain zero or one double bond and one, two, or three heteroatoms selected from the group consisting of O, N, and S. 6-membered rings contain zero, one, or two double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. 7- and 8-membered rings contain zero, one, two, or three double bonds and one, two, or three heteroatoms selected from the group consisting of O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithianyl, 1,3-dimethylpyrimidine-2,4(1H,3H)-dione, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, oxazolidin ... These include aryl, oxetanyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, 1,2-thiazinanyl, 1,3-thiazinanyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidethiomorpholinyl (thiomorpholinesulfone), thiopyranyl, and trithianyl. A bicyclic heterocycle is a monocyclic heterocycle fused to a phenyl group, or a monocyclic heterocycle fused to a monocyclic cycloalkyl, or a monocyclic heterocycle fused to a monocyclic cycloalkenyl, or a monocyclic heterocycle fused to a monocyclic heterocycle, or a spiro heterocyclic group, or a bridged monocyclic heterocycle ring system in which two non-adjacent atoms of the ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms, or by an alkenylene bridge of 2, 3, or 4 carbon atoms.Representative examples of bicyclic heterocycles include, but are not limited to, benzopyranyl, benzothiopyranyl, chromanyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzothienyl, 2,3-dihydroisoquinoline, 2-azaspiro[3.3]heptan-2-yl, azabicyclo[2.2.1]heptyl (including 2-azabicyclo[2.2.1]hept-2-yl), 2,3-dihydro-1H-indolyl, isoindolinyl, octahydrocyclopenta[c]pyrrolyl, octahydropyrrolopyridinyl, and tetrahydroisoquinolinyl. Examples of tricyclic heterocycles include bicyclic heterocycles fused to a phenyl group, or bicyclic heterocycles fused to a monocyclic cycloalkyl, or bicyclic heterocycles fused to a monocyclic cycloalkenyl, or bicyclic heterocycles fused to a monocyclic heterocycle, or bicyclic heterocycles in which two non-adjacent atoms of the bicyclic ring are linked by an alkylene bridge of 1, 2, 3, or 4 carbon atoms, or an alkenylene bridge of 2, 3, or 4 carbon atoms. Examples of tricyclic heterocycles include, but are not limited to, octahydro-2,5-epoxypentalene, hexahydro-2H-2,5-methanocyclopenta[b]furan, hexahydro-1H-1,4-methanocyclopenta[c]furan, aza-adamantane (1-azatricyclo[3.3.1.1. 3,7 ]decane), and oxa-adamantane (2-oxatricyclo[3.3.1.1 3,7 ]decane. The monocyclic, bicyclic, and tricyclic heterocycles are connected to the parent molecular moiety through any carbon atom or any nitrogen atom contained within the ring, and can be unsubstituted or substituted.

[0060] As used herein, the term "hydroxyl" means an --OH group.

[0061] As used herein, the term "hydroxyalkyl" refers to a group, as defined herein, in which one, two, three, four, five, six, seven, or eight hydrogen atoms are replaced by a hydroxyl group. It means an alkyl group as defined herein.

[0062] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkyl or cycloalkyl) is indicated by the prefix "C x ~C y -" or "C x~y (where x is the minimum number of carbon atoms in the substituent and y is the maximum number). Thus, for example, "C1-C3-alkyl" and "C 1~3 "Alkyl" refers to an alkyl substituent containing 1 to 3 carbon atoms. x ~C y -" and "C x~y " are used interchangeably and have the same meaning.

[0063] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkyl or cycloalkyl) is indicated by the prefix "C x ~C y -" (where x is the minimum number of carbon atoms in the substituent and y is the maximum number). Thus, for example, "C1-C3-alkyl" refers to an alkyl substituent containing from 1 to 3 carbon atoms.

[0064] The term "substituted" refers to a group that may be further substituted with one or more non-hydrogen substituents. Substituents include, but are not limited to, halogen, =O, =S, cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, ketone, amide, carbamate, and acyl.

[0065] The term "tetrazine" refers to a substituted or unsubstituted aromatic cyclic group of two carbon atoms and four nitrogen atoms having a single ring with three double bonds. Examples of tetrazine groups include 1,2,3,4-tetrazine and 1,2,4,5-tetrazine. As used herein, 1,2,4,5-tetrazine is referred to as the "Tz" group.

[0066] The term "selectively delivering" refers to the delivery of an agent (e.g., payload) to an organ or tissue (or portion thereof) requiring treatment or diagnosis without significant binding to other non-target organs or tissues (or portions thereof).

[0067] The term "payload" refers to an agent for delivery to a target site in a subject. The payload includes a therapeutic agent.

[0068] The term "therapeutic agent" refers to an agent capable of treating and / or ameliorating a medical condition or disease in a subject, or one or more symptoms thereof. The therapeutic agent of the present disclosure also includes prodrug forms of the therapeutic agent.

[0069] The term "diagnostic agent" refers to an agent that aids in the diagnosis of a medical condition or disease. Exemplary diagnostic agents include imaging agents such as paramagnetic agents, optical probes, and radionuclides. Paramagnetic agents are imaging agents that are magnetic under an externally applied field. Examples of paramagnetic agents include, but are not limited to, iron particles, including iron nanoparticles and iron microparticles. Optical probes are fluorescent compounds that can be detected by excitation with radiation at one wavelength and detection with radiation at a second, different wavelength. Optical probes of the present disclosure include, but are not limited to, Cy5.5, Alexa680, Cy5, DiD (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate) and DiR (1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine perchlorate). Other optical probes include quantum dots. A radionuclide is an element that undergoes detectable radioactive decay. Radionuclides useful in embodiments of the present disclosure include, but are not limited to: 3 H, 11 C. 13 N, 18 F, 19 F, 60 Co, 64 Cu, 67 Cu, 68 Ga, 82 Rb, 90 Sr, 90 Y, 99 Tc, 99m Tc, 111 In, 123 I, 124 I, 125 I, 129 I, 131 I, 137 Cs, 177 Lu, 186 Re, 188 Re, 211 At, Rn, Ra, Th, U, Pu and 241 Am is one example.

[0070] The term "targeting agent" refers to a chemical or biological agent that specifically binds to a target (e.g., a targeted organ or tissue), thereby forming a stable association between the targeting agent and a particular target. "Stably associated" or "stable association" means that a moiety binds to or otherwise associates with another moiety or structure under standard physiological conditions. Bonds can include covalent bonds and non-covalent interactions, including, but not limited to, ionic bonds, hydrophobic interactions, hydrogen bonds, van der Waals forces (e.g., London dispersion forces), dipole-dipole force interactions, and the like. A targeting agent may be a member of a specific binding pair, including, but not limited to, a member of a receptor / ligand pair; a ligand-binding portion of a receptor; a member of an antibody / antigen pair; an antigen-binding fragment of an antibody; a hapten; a member of a lectin / carbohydrate pair; a member of an enzyme / substrate pair; biotin / avidin; biotin / streptavidin; digoxin / antidigoxin; a member of a DNA or RNA aptamer binding pair; a member of a peptide aptamer binding pair, and the like. Targeting agents include ligands that specifically bind (or substantially specifically bind) to a particular clinically relevant target receptor or cell surface target. Ligands may be antibodies, peptides, nucleic acids, phages, bacteria, viruses, or other molecules that have a specific affinity for a target receptor or cell surface target. Examples of receptors and cell surface targets include, but are not limited to, LT4-human mAb against PD-1, CTLA-4, HER2 / neu, HER1 / EGFR, VEGFR, BCR-ABL, SRC, JAK2, MAP2K, EML4-ALK, BRAF V600E, 4-1BB, GITR, GSK3β, inhibitory immune checkpoint receptor immunoglobulin-like transcript 4 (ILT4); leukocyte immunoglobulin-like receptor superfamily B member 2, LILRB2, lymphocyte immunoglobulin-like receptor 2, LIR2, monocyte / macrophage immunoglobulin-like receptor 10, MIR-10, CD85d, or other cell receptors or cell surface targets.

[0071] The term "targeted organ or tissue" refers to an organ or tissue that is targeted for delivery of a payload. Exemplary organs and tissues that may be targeted include those that can be targeted by chemical or biological targeting agents, as well as those that cannot be targeted by chemical or biological targeting agents.

[0072] The term "implanting" refers to surgical implantation into a subject's body.

[0073] The term "contacting" or "contacting" refers to the process of bringing at least two different species into contact so that they can interact with one another, for example, in a non-covalent or covalent interaction or binding reaction. However, it should be understood that the resulting complex or reaction product may be produced directly from the interaction or reaction of the added reagents, or from an intermediate (which may be produced in the contact mixture) derived from one or more of the added reagents or moieties.

[0074] The term "binding agent" refers to an agent having a functional group capable of forming a covalent bond with a complementary functional group of another binding agent in a biological environment. The binding may also be referred to as a bioconjugation reaction. Binders include bioorthogonal binders, which are binders that have a bioorthogonal functional group. The bioorthogonal functional group of a bioorthogonal binder selectively reacts with the complementary bioorthogonal functional group of another bioorthogonal binding partner. The selective reaction of the bioorthogonal binding partners with each other can minimize side reactions with other binders, biological compounds, or other non-complementary bioorthogonal binders or non-complementary bioorthogonal functional groups. Bioorthogonal functional groups of bioorthogonal binders include, but are not limited to, azides and alkynes for triazole formation by click chemistry, trans-cyclooctene (TCO) and tetrazine (Tz) (e.g., 1,2,4,5-tetrazine). Binders useful in the present disclosure may be highly reactive with the corresponding binders such that the reaction is rapid.

[0075] The term "functionalized" refers to a moiety having a functional group attached, for example, a binding agent functional group (eg, a bioorthogonal functional group) attached.

[0076] The term "administering" refers to any suitable route of administration to a subject, including, but not limited to, oral administration, administration as a suppository, topical contact, parenteral, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, intrathecal administration, or implantation of a sustained release device, such as a mini-osmotic pump, to a subject.

[0077] As used herein, "parenteral" refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.

[0078] The term "leaving group" refers to an atom (or group of atoms) with electron-withdrawing capability that can be displaced as a stable species, thereby accepting a bonding electron. Examples of suitable leaving groups include halides (e.g., Br, Cl, I), sulfonate esters (e.g., triflate, mesylate, tosylate, and brosylate), and nitrophenols.

[0079] The terms "pharmacologically effective amount" and "therapeutically effective amount" refer to an amount of a compound sufficient to treat a specified disorder or disease, or one or more symptoms thereof, and / or to prevent or reduce the risk of onset or recurrence of a disorder or disease, or one or more symptoms thereof. With respect to tumorigenic proliferative disorders, a pharma- ceutically effective or therapeutically effective amount includes, inter alia, an amount sufficient to shrink a tumor or reduce the rate of tumor growth.

[0080] As used herein, "subject", "patient" or "organism" includes humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). Typical subjects to which the agents of the present disclosure can be administered may include mammals, particularly primates, especially humans. For veterinary applications, suitable subjects may include, for example, livestock animals such as cattle, sheep, goats, cows, pigs; poultry such as chickens, ducks, geese, turkeys; and pets such as dogs and cats. For diagnostic or research applications, suitable subjects may include mammals, for example, rodents (e.g., mice, rats, hamsters), rabbits, primates, and pigs, such as inbred pigs.

[0081] The term "treating" or "treatment" as used herein means treating or managing a disease or condition or symptom thereof in a patient, such as a mammal (especially a human), which may include: (a) ameliorating the disease or condition or symptom thereof, e.g., eliminating or causing the regression of the patient's disease or condition or symptom thereof; (b) inhibiting the patient's disease or condition or symptom thereof, e.g., by slowing or arresting the onset of the disease or condition or symptom thereof; or (c) preventing the patient's disease or condition or symptom thereof. This includes alleviating the condition or symptoms thereof.

[0082] The term "physiological conditions" is meant to encompass conditions compatible with living cells, eg, primarily aqueous conditions of temperature, pH, salinity, and the like, compatible with living cells.

[0083] For the compounds described herein, the groups and substituents may be selected according to the allowed valences of the atoms and substituents, such that the selection and substitution give stable compounds, e.g., compounds that do not spontaneously undergo transformation by rearrangement, cyclization, elimination, and the like.

[0084] When a range of values ​​is described, it is understood that all intervening values ​​between the upper and lower limits of that range (to one-tenth of the unit of the lower limit unless the context clearly requires otherwise), and other stated or intervening values ​​within this description are encompassed within the scope of the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and may also be encompassed within the invention, subject to any specifically excluded limit in this described range. Where the described range includes one or both limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0085] When numerical ranges are recited herein, each of the intervening values ​​therebetween, to the same degree of precision, is expressly contemplated, for example, in the range of 6 to 9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and in the range of 6.0 to 7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0086] It is understood that certain features of the invention that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described for brevity in the context of a single embodiment may also be provided separately or in any suitable subcombination. Any combination of the embodiments of the invention is specifically embraced by the present invention and is disclosed herein exactly as if each and every combination were individually and expressly disclosed, to the extent that such combinations include subject matter that is, for example, a stable compound (i.e., a compound that can be made, isolated, characterized, and tested for biological activity). In addition, any subcombination of the various embodiments and elements thereof (e.g., elements of chemical groups listed in the embodiments that illustrate such variations) is also specifically embraced by the present invention and is disclosed herein exactly as if each and every such subcombination were individually and expressly disclosed herein.

[0087] 2. Composition A. trans-Cyclooctene Functionalized Payloads The trans-cyclooctene functionalized payloads of the present disclosure are compounds of formula (I) (D, R 1a , R 1b , L 1 , m and p are as defined herein.

[0088] The compound of formula (I) can be prepared by reacting a compound of formula (IA) (D, R 1a , R 1b , L 1 , m and p are as defined herein. [ka]

[0089] In the compounds described herein, R 1a and R 1b may be hydrogen.

[0090] In the compounds described herein, R 1a is C 1~4 R is alkyl; 1b are as follows: C(O)OH, C(O)OC 1~4 Alkyl, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )CHR 1e C(O)OC 1~4 Alkyl, C(O)N(R 1c )-C 1~6 Alkylene-COH and C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 R may be selected from the group consisting of alkyl. 1b is further divided into C(O)OH, C(O)N(R 1c )CHR 1e COH and C(O)N(R 1c )CH2CO2H.

[0091] In the compounds described herein, R 1emay be -CH2CO2H, -CH2CH2CO2H, -CH2CONH2, -CH2CH2CONH2, -CH2OH, or -CH(CH3)OH; or R 1e -C 1~4 alkylene-COH; or R 1e is -CH2CO2H.

[0092] In the compounds described herein, R 1a may be hydrogen.

[0093] In the compounds described herein, R 1a is C 1~4 It may be alkyl.

[0094] In the compounds described herein, R 1a may be CH3.

[0095] In the compounds described herein, R 1b may be hydrogen.

[0096] In the compounds described herein, R 1b is C(O)N(R 1c )-C 1~6 It may be alkylene-COH.

[0097] In the compounds described herein, R 1b is C(O)N(R 1c )CH2CO2H.

[0098] In the compounds described herein, R 1b may be C(O)OH.

[0099] In the compounds described herein, R 1c may be hydrogen.

[0100] The trans-cyclooctene functionalized payload of the present disclosure is a compound of formula (IB), or a pharma- ceutically acceptable salt thereof: [ka] Including, In the formula, D 1 is, at each occurrence, a payload independently selected from an anticancer drug payload, a microbial immunosuppressant drug payload, a restenosis inhibitor drug payload, an antibiotic drug payload, an antifungal drug payload, an antiviral drug payload, an anti-inflammatory drug / anti-arthritic drug payload, a corticosteroid drug payload, and an immunosuppressant drug payload; R 1a , R 1b , L 1 , p, and m are as defined herein for formula (IB). For example, p can be 0; m is 1; L 1 teeth, [ka] It is.

[0101] In some embodiments, the anticancer drug is doxorubicin.

[0102] The trans-cyclooctene functionalized payload of the present disclosure has formula (II-A), 1 , R 1A , R 1B , L 1 , L 2 , m and p are as defined herein.

[0103] The compound of formula (II-A) is represented by the formula (II-A') (wherein D 1 , R 1A , R 1B , L 1 , L 2 , m and p are as defined herein. [ka]

[0104] In some embodiments, R 1B is as follows:G1 , OH, -NR 1c -C 1~4 Alkylene -G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, and -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1e is -CH2CO2H, -CH2CH2CO2H, -CH2CONH2, -CH2CH2CONH2, -CH2OH, or -CH(CH3)OH; R 1f is hydrogen or CH2CO2H; where n, G 1 and R 1c is as defined herein.

[0105] In some embodiments, R 1A is C 1~4 R is alkyl; 1B is G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, and -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1e -C 1~4 alkylene-COH; R 1f is hydrogen or C 1~4 Alkylene-COH; G 1 is a 4-8 membered monocyclic heterocyclyl containing a first nitrogen and optionally one additional heteroatom selected from nitrogen, oxygen, and sulfur; G1 is attached to the first nitrogen, and optionally, C 1~4 Alkyl, C 1~4 Haloalkyl, halo, cyano, OH, -OC 1~4 substituted with 1 to 4 substituents independently selected from the group consisting of alkyl, alkyl, and oxo; n is 0, 1, or 2, where R 1c and R 1d is as defined herein.

[0106] In some embodiments, R 1A is CH3; R 1e is -CH2CO2H; R 1f is hydrogen or CH2CO2H; G 1 is attached via a ring nitrogen atom, and optionally 1~4 Alkyl, C 1~4 Haloalkyl, halo, cyano, OH, -OC 1~4 piperazinyl (e.g., piperazin-1-yl), morpholinyl (e.g., morpholin-4-yl), piperidinyl (e.g., piperidin-1-yl), azepanyl (e.g., azepan-1-yl), or pyrrolidinyl (e.g., pyrrolidin-1-yl), substituted by 1 to 4 substituents independently selected from the group consisting of alkyl, and oxo.

[0107] In some embodiments, L 2 is -C(O)-.

[0108] In some embodiments, R 1B are OH, -N(H)CH2CO2H, -N(H)CHR 1e CO2H, -N(H)CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2, and -N(CH2CO2H)-CH2CH2-N(CH2CO2H)2; R 1e is -CH2CO2H.

[0109] In some embodiments, L 2 is -C(O)-; R 1Ais C 1~4 R is alkyl; 1B OH, -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1~6 Alkylene -COH or -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-COH)-C 2~4 Alkylene) n -N(C 1~4 alkylene-COH; R 1c is hydrogen or C 1~4 R is alkyl; 1e -C 1~4 alkylene-COH; R 1f is hydrogen or C 1~4 Alkylene-COH; m, n, p, D 1 , and L 1 is as defined herein.

[0110] In some embodiments, L 2 is -C(O)-; R 1A is C 1~4 R is alkyl; 1B OH, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, or -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1c is hydrogen or C 1~4 R is alkyl; 1e -C 1~4 alkylene-COH; R 1f is hydrogen or C 1~4 Alkylene-COH; m, n, p, D 1 , and L 1 is as defined herein.

[0111] In another embodiment, L 2 is -C(O)-; R 1Ais CH3; R 1B OH, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, or -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1e is -CH2CO2H, -CH2CH2CO2H, -CH2CONH2, -CH2CH2CONH2, -CH2OH, or -CH(CH3)OH; R 1f is hydrogen or CH2CO2H; R 1c is hydrogen or CH3; m, n, p, D 1 , and L 1 is as defined herein.

[0112] In yet another embodiment, L 2 is -C(O)-; R 1A is CH3; R 1B are OH, -N(H)CH2CO2H, -N(H)CHR 1e CO2H, -N(H)CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2 or -N(CH2CO2H)-CH2CH2-N(CH2CO2H)2; R 1e is -CH2CO2H; m, n, p, D 1 , and L 1 is as defined herein.

[0113] The trans-cyclooctene functionalized payload of the present disclosure has formula (III-A): 1 , R 1B , L 1 , L 2 , m and p are as defined in formulas (II-A) and (II-A'). [ka]

[0114] In the compounds described herein, the linker L 1 may have 1-100 linking atoms and may include ethylene-oxy groups, amines, esters, amides, carbamates, carbonates, and ketone functionalities. For example, the linker may have 1-50 linking atoms, or 5-50 linking atoms, or 10-50 linking atoms.

[0115] The linker may be a non-releasable linker. A non-releasable linker forms a bond between at least two moieties that is not significantly disrupted (e.g., the covalent bond in the linker remains intact and is not cleaved) under the conditions in which the composition employing the non-releasable linker is used.

[0116] The linker may be a releasable linker. A releasable linker forms a bond between at least two moieties that can be broken under release conditions (e.g., one or more bonds in the linker can be cleaved) such that the moieties are no longer bound to each other. A releasable linker can have bonds between the moieties that are broken when the releasable linker is exposed to release conditions, such as, but not limited to, light, heat, sound, a releasing agent (e.g., a chemical releasing agent (e.g., an acid, a base, an oxidizing agent, a reducing agent), a solvent, an enzyme, etc.), a combination thereof, etc. In some embodiments, a releasable linker forms a bond between the moieties that can be broken under release conditions ... In some cases, the application of an external stimulus or contact with a release condition is not required to break the bond. For example, a releasable linker may include one or more labile bonds or functional groups in the linker that can be spontaneously cleaved without contact with an external stimulus or release condition, thereby releasing the payload from the support composition. As mentioned above, an example of a bond or functional group that can be spontaneously cleaved includes, but is not limited to, carbamate, which releases carbon dioxide upon spontaneous cleavage. The functionalized payload of the present disclosure that includes a releasable linker can facilitate the delivery of the payload to a target site in a subject.

[0117] In some cases, the payload can be released as described above by contacting the releasable linker with a release condition. The release condition can be target specific, for example, a release condition that is applied directly to a desired target location in a subject (e.g., a target location where the therapeutic support composition is present). In some embodiments, the release condition can be non-specific, for example, by exposure of the releasable linker to extracellular mechanisms (e.g., low pH in tumor tissue, hypoxia, enzymes, etc.). In other cases, the release of the payload can be achieved via intracellular (e.g., lysosomal) release mechanisms (e.g., glutathione, proteases (e.g., cathepsins), catabolism, etc.). In these cases, the therapeutic support composition can be internalized into the cell and then exposed to the release conditions present within the cell. The intracellular release condition (e.g., glutathione, cathepsins, etc.) can cause the release of the payload from the therapeutic support composition, allowing the payload to disperse from the cell and provide a therapeutic effect to neighboring cells. Examples of these types of releasable linkers include, but are not limited to, hydrazones (acid labile), peptide linkers (cathepsin B cleavable), disulfide moieties (thiol cleavable), etc. This type of release mechanism of action may facilitate the treatment of diseases or conditions such as tumors (e.g., tumors with heterologous receptor expression or tumors with poor mAb penetration).

[0118] In certain embodiments, the linker between the payload and the trans-cyclooctene is an immolative linker.

[0119] In certain embodiments, the linker between the payload and trans-cyclooctene is a pH-tunable linker.

[0120] In some cases, the therapeutic agent is covalently attached to the linker via an amide bond; for example, the therapeutic agent may be an amine-containing therapeutic agent to attach the therapeutic agent to a carbonyl group of the linker, or in other cases, the therapeutic agent may be a carboxyl-containing therapeutic agent to attach the therapeutic agent to an amine group of the linker. In some cases, the therapeutic agent and the linker together form a carbamate group; for example, the therapeutic agent may be an amine-containing therapeutic agent to attach the therapeutic agent to an acyloxy group of the linker. In some cases, the therapeutic agent and the linker together form a carbonate group; for example, the therapeutic agent may be a hydroxyl-containing therapeutic agent to attach the therapeutic agent to an acyloxy group of the linker.

[0121] For example, in the compounds described herein, L 1 teeth, [ka] or -O-; Here, L 3 is a bond or C 1~6 alkylene; L 4 is the bond, -NHN:-N(R 10 )-C 2~6 Alkylene-N(R 11 )-, -N(R 12 )-C 2~3 Alkylene -N(R 13 )C(O)-, -N(R 10 )-C 1~6 Alkylene-C(O)NHN: -NHNHC(O)C 1~6 Alkylene -C(O)NHN: -CH(NHC(O)R 14 ) C 1~4 Alkylene-SSC 1~4 Alkylene -OC(O)-, -NHNHC(O)CH(NHC(O)R 15 )CH2C(O)-, -C 1~6 Alkylene-CH(G x )OC(O)-, [ka] ;R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , and R 19 are each independently hydrogen or C 1~4 R is alkyl; 16 is hydrogen, C 1~4 Alkyl, -C 1~4 Alkylene -OH, -C 1~4 Alkylene-OC 1~4 Alkyl, -C 1~4 Alkylene-COH or -C 1~4 Alkylene-CONH2; R 17 is independently at each occurrence hydrogen or -CH2OC(O)-; G x Optionally, halogen, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 It is phenyl substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy, cyano, and nitro.

[0122] In the compounds described herein, m can be 1. When m is 1, [ka] teeth, [ka] [ka] wherein R 18 is independently at each occurrence hydrogen or -CH2OC(O)NHD'; R D is a hydrogen or C on the nitrogen atom of the payload 1~4is alkyl; and D' is a payload moiety (eg, a cyclic dinucleotide payload moiety, an imidazo[4,5-c]quinolin-4-amine payload moiety, a TLR agonist payload moiety, a STING agonist payload moiety).

[0123] In the compounds described herein, [ka] teeth, [ka] where D' is a cyclic dinucleotide payload moiety.

[0124] If m is 1, then [ka] teeth, [ka] [ka] wherein R 18 Each occurrence is independently hydrogen or -CH2OC(O)NHD 1a ;R D is a hydrogen or C on the nitrogen atom of the payload 1~4 Alkyl; D 1a is a payload moiety (e.g., an anti-cancer payload moiety).

[0125] Those skilled in the art will recognize that the payload D / D 1 It will be appreciated that the D / D from the compounds herein does not refer to the payload molecule itself, but rather to the portion of the payload molecule attached to the linker. 1 Upon release of the payload, the payload itself is released.

[0126] As used herein, a "payload moiety" refers to that nucleophilic group attached to a linker, e.g., NH, NC 1~4 or its electrophilic group attached to the linker, e.g., C(O), i.e., payload D / D excluding the remainder of the payload 1 For example, the formula [ka] The compound is [ka] Compounds such as; [ka] teeth, [ka] Including compounds such as; [ka] teeth, [ka] Compounds such as: [ka] The compound is [ka] D 1 H, NH2-D 1a ,HOOC-D 1a , or the release of HO-D' releases the payload itself.

[0127] In the compounds described herein, p can be 0.

[0128] In the compounds described herein, m can be 2 or 3. In some embodiments, m is 2; [ka] teeth, [ka] It is.

[0129] In some embodiments, m is 2; [ka] teeth, [ka] It is.

[0130] D 1may be a drug payload selected from an anti-cancer drug payload, a microbial immunosuppressant payload, or a restenosis inhibitor payload. The anti-cancer drug may be one or more selected from the following: methotrexate, purines, pyrimidines, plant alkaloids, epothilones, triptolide compounds, antibiotics (particularly actinomycin D), hormones, and antibodies. Among the plant alkaloids, mention may be made of paclitaxel, doxorubicin, maytansine, auristatins, calicheamicins, duocarmycins, tubulysins, and camptothecins, among others. The microbial immunosuppressant may be one or more selected from the following: cyclosporin A, tacrolimus and its analogs, despergualin, mycophenolic acid esters, rapamycin and its derivatives, FR-900520 substances from Streptomyces strains, FR-900523 substances from Streptomyces strains, daclizumab, pentanamides, canglemycin C, spergualin, prodigiosin-25C, tranilast, myriocin, cyclosporin C, bredinin, mycophenolic acid, brefeldin A, and ketosteroids. The restenosis inhibitor may be one or more selected from the following: batimastat, metalloproteinase inhibitors, 17β-estradiol, NO donors, 2-chlorodeoxyadenosine, 2-deoxycoformycin, fingolimod, sodium mycophenolate, ISA. TX The agent may be one or more selected from 247 (cyclosporine A derivative), elsibucole, daclizumab, basiliximab, antithymocyte globulin, everolimus, methotrexate, neoral, cyclophosphamide, brequinar sodium, leflunomide and mizoribine.

[0131] Exemplary anticancer drugs include, but are not limited to, the following: abiraterone acetate, abitrexate (methotrexate), abraxane (paclitaxel albumin-stabilized nanoparticle formulation), ABVD, ABVE, ABVE-PC, AC, AC-T, adcetris (brentuximab vedotin), ADE, ado-trastuzumab emtansine, adriamycin (doxorubicin hydrochloride), adrsil (fluorouracil), afatinib dimaleate, afinitor (everolimus), aldara (imiquimod), aldes Leukin, Alemtuzumab, Alimta (Pemetrexed Disodium), Aloxi (Palonosetron Hydrochloride), Ambochlorin (Chlorambucil), Aminolevulinic Acid, Anastrozole, Aprepitant, Aredia (Pamidronate Disodium), Arimidex (Anastrozole), Aromasin (Exemestane), Alanone (Nelarabine), Arsenic Trioxide, Arzera (Ofatumumab), Asparaginase Erwinia c hrysanthemi), Avastin (bevacizumab), axitinib, azacitidine, BEACOPP, bendamustine hydrochloride, BEP, bevacizumab, bexarotene, Bexar (tositumomab and I-131 iodine tositumomab), bicalutamide, bleomycin, bortezomib, Bosulif (bosutinib), bosutinib, brentuximab vedotin, busulfan, Busulfex (busulfan), cabazitaxel, cabozantinib-S-malate, CAF, Campus (alemtuzumab), Camptosa Taxol (irinotecan hydrochloride), capecitabine, CAPOX, carboplatin, carboplatin-taxol, carfilzomib, Casodex (bicalutamide), CeeNU (lomustine), Cerbidine (daunorubicin hydrochloride), Cervarix (recombinant HPV bivalent vaccine), cetuximab, chlorambucil, chlorambucil-prednisone, CHOP, cisplatin, Clafen (cyclophosphamide), clofarabine, Clofarex (clofarabine), Chloral (clofarabine), CMF, rice Triq (cabozantinib-S-malate), COPP, COPP-ABV, Cosmegen (dactinomycin), crizotinib, CVP, cyclophosphamide, Cyfos (ifosfamide), cytarabine, cytarabine, liposomal, Cytosar-U (cytarabine), Cytoxan (cyclophosphamide), dabrafenib, dacarbazine, Dacogen (decitabine), dactinomycin, dasatinib, daunorubicin hydrochloride, decitabine, degarelix, denileukin diftitox, denosumab, DepoCyt (lis DepoFoam (liposomal cytarabine), Dexrazoxane hydrochloride, Docetaxel, Doxil (doxorubicin hydrochloride liposomal), Doxorubicin hydrochloride, Doxorubicin hydrochloride liposomal, Dox-SL (doxorubicin hydrochloride liposomal), DTIC-Dome (dacarbazine), Efudex (fluorouracil), ERYTECH (rasburicase), Elence (epirubicin hydrochloride), Eloxatin (oxaliplatin), Eltrombopag Olamine, Emend (aprepitant), Enzalutamide, Epirubicin hydrochloride, EPOCH, Erbitux (cetuximab), Eribulin mesylate,Erivage (vismodegib), erlotinib hydrochloride, Erwinase (asparaginase-erwinia chrysanthemi), Etopofos (etoposide phosphate), etoposide, etoposide phosphate, Everset (doxorubicin hydrochloride liposome), everolimus, Evista (raloxifene hydrochloride), exemestane, Fareston (toremifene), Faslodex (fulvestrant), FEC, Femara (letrozole), filgrastim, Fludara (fludarabine phosphate), fludarabine phosphate, Fluoroplex (fluorouracil), Fluorouracil, Forex (methotrexate), ForexPFS (methotrexate), Forfiri, Forfiri-bevacizumab, Forfiri-cetuximab, Forfirinox, Forfox (leucovorin, fluorouracil, oxaliplatin), Forothin (pralatrexate), FU-LV, fulvestrant, Gardasil (recombinant HPV quadrivalent vaccine), Gazyva (obinutuzumab), gefitinib, gemcitabine hydrochloride, gemcitabine-cisplatin, gemcitabine-oxaliplatin, Gemtu Zumab ozogamicin, Gemzar (gemcitabine hydrochloride), Gilotrif (afatinib dimaleate), Glivec (imatinib mesylate), glucarpidase, goserelin acetate, Halaven (eribulin mesylate), Herceptin (trastuzumab), HPV bivalent vaccine (recombinant), HPV quadrivalent vaccine (recombinant), Hycamtin (topotecan hydrochloride), Hyper CVAD, ibritumomab tiuxetan, ibrutinib, ICE, Iclusig (ponatinib hydrochloride), Ifex (Ifosf) imatinib mesylate, Imbruvica (ibrutinib), imiquimod, Inlyta (axitinib), Intron A (recombinant interferon alpha-2b), iodine-131 tositumomab and tositumomab, ipilimumab, Iressa (gefitinib), irinotecan hydrochloride, Istodax (romidepsin), ixabepilone, Ixempra (ixabepilone), Jakafi (ruxolitinib phosphate), Jevtana (cabazitaxel), Kadcyla (Ado-trastuzumab emtansine),, Keoxifene (raloxifene hydrochloride), Kepivans (palifermin), Kyprolis (carfilzomib), lapatinib ditosylate, lenalidomide, letrozole, leucovorin calcium, Leukeran (chlorambucil), leuprolide acetate, Levran (aminolevulinic acid), Lympholidine (chlorambucil), LipoDox (doxorubicin hydrochloride liposomal), liposomal cytarabine, lomustine, Lupron (leuprolide acetate), Lupron Depot (leuprolide acetate), Lupron Depot-P ed (leuprolide acetate), Leupro Depot-3 months (leuprolide acetate), Leupro Depot-4 months (leuprolide acetate), Marquibo (vincristine sulfate liposomal), Matulane (procarbazine hydrochloride), mechlorethamine hydrochloride, Megace (megestrol acetate), megestrol acetate, Mekinist (trametinib), mercaptopurine, mesna, Mesnex (mesna), Metazolastone (temozolomide), methotrexate, methotrexate LPF (methotrexate), Me Mexate (methotrexate), Mexate-AQ (methotrexate), Mitomycin C, Mitoditrex (mitomycin C), MOPP, Mozovir (plelixafor), Mustalgen (mechlorethamine hydrochloride), Mutamycin (mitomycin C), Myleran (busulfan), Mirosal (azacitidine), Milotarg (gemtuzumab ozogamicin), nanoparticle paclitaxel (paclitaxel albumin-stabilized nanoparticle formulation), Navelbine (vinorelbine tartrate), Nelarabine, Neosal (cyclophosphamide), Neupogen ( Filgrastim), Nexavar (sorafenib tosylate), nilotinib, Nolvadex (tamoxifen citrate), Nplate (romiplostim), obinutuzumab, ofatumumab, omacetaxine mepesuccinate, Oncaspar (pegaspargase), Ontac (denileukin diftitox), OEPA, OPPA, oxaliplatin, paclitaxel, paclitaxel albumin-stabilized nanoparticle formulation, palifermin, palonosetron hydrochloride, pamidronate disodium, panitumumab, Paraplat (carboplatin),Paraplatin (carboplatin), pazopanib hydrochloride, pegaspargase, peginterferon alfa-2b, PEG-Intron (peginterferon alfa-2b), pemetrexed disodium, Perjeta (pertuzumab), pertumumab, Platinol (cisplatin), Platinol-AQ (cisplatin), plerixafor, pomalidomide, Pomalyst (pomalidomide), ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, Proleukin (aldesleukin), Prolia (denosumab), Promacta (eltrombopag olamine), Provenge (sipuleucel-T), Purinetol (mercaptopurine), radium-223, dichloride, raloxifene hydrochloride, rasburicase, R-CHOP, R-CVP, recombinant HPV bivalent vaccine, recombinant HPV quadrivalent vaccine, recombinant interferon alpha-2b, regorafenib, Revlimid (lenalidomide), Rheumatrex (methotrexate), Rituxan (rituximab), rituximab, romidespin, romiplostim, rubidomycin (daunorubicin hydrochloride), ruxolitinib phosphate, sclero Sole Intraplural Aerosol (talc), Sipuleucel-T, Sorafenib Tosylate, Sprycel (dasatinib), Stanford V, Sterile Talc Powder (talc), Steritalc (talc), Stivarga (regorafenib), Sunitinib Malate, Stent (sunitinib malate), Sylatron (pegylated interferon alpha-2b), Synovir (thalidomide), Synribo (omacetaxine mepesuccinate), Tafinlar (dabrafenib), Talc, Tamoxifen Citrate, Tarabin PFS (cytarabine), Tarceva (erlotinib hydrochloride), Targretin (bexarotene), Tasigna (nilotinib), Taxol (paclitaxel), Taxotere (docetaxel), Temodar (temozolomide), temozolomide, temsirolimus, thalidomide, Thalomid (thalidomide), Toposar (etoposide), topotecan hydrochloride, toremifene, Torisel (temirolimus), tositumomab and I131 iodine tositumomab, Totect (dexrazoxane hydrochloride), trametinib, trastuzumab, Treanda (bendamustine hydrochloride),Trisenox (arsenic trioxide), Tykerb (lapatinib ditosylate), vandetanib, VAMP, Vectibix (panitumumab), VelP, Velban (vinblastine sulfate), Velcade (bortezomib), Versal (vinblastine sulfate), vemurafenib, Bepcid (etoposide), Viadur (leuprolide acetate), Vidaza (azacitidine), vinblastine sulfate, Vincersar PFS (vincristine sulfate), vincristine sulfate, vincristine sulfate liposomal, vinorelbine tartrate, vismodegib, boraxase (glucarpidase), vorinostat, Votrient (pazopanib hydrochloride), Wellcovorin (leucovorin) Calcium, Xalkori (crizotinib), Xeloda (capecitabine), Xelox, Xgeva (denosumab), Xofigo (radium-223 dichloride), Xtandi (enzalutamide), Yervoy (ipilimumab), Zaltrap (Ziv-aflibercept), Zelboraf (vemurafenib), Zevalin (ibritumomab tiuxetan), Zinecard (dexrazoxane hydrochloride), Ziv-aflibercept, Zoladex (goserelin acetate), zoledronic acid, Zolinza (vorinostat), Zometa (zoledronic acid), and Zytiga (abiraterone acetate).

[0132] In certain embodiments, D 1The drug payloads of are PBD dimer, calicheamicin, speromycin, tubulysin B, rhizoxin, dolastatin, didemnin B, camptothecin, CBI, temsirolimus, actinomycin D, epothilone B, taxol, cryptophycin, SN38, velcade, bruceanthin, DAVLBH, DM1, phyllanthoside, alimta, T2 toxin, MMC, banta vantalanib, vinorelbine, brefeldin, sunitinib, daunomycin, semaxanib, tarceva, iressa, irinotecan, LY-541503, geldanomycin, gemcitabine, methotrexate, gleevec, topotecan, bleomycin, doxorubicin, cisplatin, N-mustards, etoposide, or 5-FU.

[0133] In certain embodiments, the anti-cancer drug is an anthracycline. In certain embodiments, the anti-cancer drug is a taxane. In certain embodiments, the anti-cancer drug is gemcitabine. In certain embodiments, the anti-cancer drug is doxorubicin. In certain embodiments, the anti-cancer drug is docetaxel. In certain embodiments, the anti-cancer drug is SN38. In certain embodiments, the anti-cancer drug is monomethyl auristatin E. In certain embodiments, D 1 The drug payload of is dexamethasone. In certain embodiments, 1 The drug payload of is celecoxib. In certain embodiments, D 1 The drug payload of D is gentamicin. 1 In some embodiments, the drug payload of D is vancomycin. 1 In some embodiments, the drug payload of D is daptomycin. 1 In some embodiments, the drug payload of D is doxorubicin. 1 In some embodiments, the drug payload of D is gemcitabine. 1 The drug payload of D is docetaxel. 1The drug payload of is cyclic adenosine monophosphate (c-AMP).

[0134] Specific [ka] As, [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] Examples include:

[0135] Any of the embodiments described herein may further comprise: [ka] but, [ka] [ka] In yet another embodiment,

[0136] Preferred compounds of formula (IB) and (II-A) are those of formula [ka] Compounds of, for example, [ka] Examples include:

[0137] Preferred compounds of formula (IB) and (II-A) are those of formula [ka] Compounds of, for example, [ka] for example, [ka] Examples include:

[0138] Preferred compounds of formula (IA) include those of formula [ka] Compound Examples include:

[0139] Payload D is a TLR agonist or a STING agonist.

[0140] TLR agonists are immunomodulatory agents. TLR-mediated signaling in response to pathogen-associated molecular patterns (PAMPs) is a sequential cascade of transcriptional regulatory events that vary depending on the TLR agonist, the cell type involved, and the pathogenicity of the antigen. Individual genes (particularly inflammatory cytokines such as IL-1 (α and β), IL-6, IL-18, TNF-C) are transiently induced, representing the ability of the innate immune system to interpret the infection and direct an appropriate response while promoting resolution (T. Ravasi, CA Wells, DA Hume, Bioessays 29, 1215 (Nov. 15, 2007); JC Roachet al., Proc Natl Acad Sci USA 104, 16245 (Oct. 9, 2007); M. Gilchrist et al., Bioessays 29, 1215 (Nov. 15, 2007); J. C Roachet al., Proc Natl Acad Sci USA 104, 16245 (Oct. 9, 2007); M. Gilchrist et al., Bioessays 29, 1215 (Nov. 15 ...Oct. 9, 2007); M. Gilchrist et al., Bioessays 29, 1215 (Oct. 9, 2007); M. Gilchrist et al., Bioessays 29, 1215 (Oct. 9, 2007); M. Gilchrist et al., Bioessays 29, 121 al., Nature 441, 173 (May 11, 2006)).

[0141] TLR agonists include, but are not limited to, TLR1 / 2 heterodimers (e.g., Pam3CSK4, i.e., trispalmitoyl CysSerLysLysLysLys), TLR3 (e.g., poly I:C, poly ICLC), TLR4 (e.g., monophosphoryl lipid A, lipopolysaccharide, GLA-SE, G100), TLR5 (e.g., flagellin), TLR2 / 6 heterodimers (e.g., diacyl lipopeptides of gram-positive bacteria, mycoplasma, and fungi), TLR7 (e.g., imidazo[4,5-c]quinolin-4-amines, e.g., imiquimod, and the like, which are incorporated by reference herein, and are not limited to any of the U.S. Pat. Nos. 5,993,333, 5,9 ... No. 4,689,338, as well as polyriboinosinic acid-polyribocytidylic acid (poly I:C), TLR3 (polyadenylic acid-polyuridylic acid (poly A:U)), TLR2 (peptidoglycan), TLR2 and TLR4 (e.g., Bacillus Calmette-Guerin (BCG)), [ka] TLR7 / 8 (e.g., loxoribine, imidazo[4,5-c]quinolin-4-amines, e.g., resiquimod (R848) and MEDI9197); [ka] TLR8 (e.g., VTX-2337) [ka] and TLR9 (e.g., CpG ODNs, e.g., ODNs D-SL01, MGN1703, CPG7909, SD-101, EMD 1201081). CpG ODNs are short synthetic single-stranded DNA molecules that contain unmethylated CpG dinucleotides in a specific sequence context (CpG motif). CpG ODNs have a partially or fully phosphorothioated (PS) backbone, in contrast to the natural phosphodiester (PO) backbone present in genomic bacterial DNA. Based on their structural features and activity on human peripheral blood mononuclear cells (PBMCs), particularly B cells and plasmacytoid dendritic cells (pDCs), three main classes of stimulatory CpG ODNs have been identified. CpG-A ODNs are characterized by a PO-centered CpG-containing palindromic motif and a PS-modified 3' poly-G tract. They induce high IFN-α production from pDCs but are weak stimulators of TLR9-dependent NF-κB signaling and inflammatory cytokine (e.g., IL-6) production. CpG-B ODNs contain an entire PS backbone with one or more CpG dinucleotides. They potently activate B cells and TLR-dependent NF-κB signaling but are weak at stimulating IFN-α secretion. CpG-C ODNs combine features of both classes A and B. They contain a complete PS backbone and a CpG-containing palindromic motif. C class CpG ODNs induce strong IFN-α production from pDCs as well as B cell stimulation.

[0142] Further TLR9 agonists are described in WO 2019 / 115402, EP 2017281, U.S. Patent No. 2019 / 0160173, U.S. Patent No. 2019 / 0151345, U.S. Patent No. 2011 / 0311518, and U.S. Patent No. 2011 / 0293565, each of which is incorporated by reference herein.

[0143] Single- and double-stranded RNA can function as TLR agonists, as described by Roers et al. in Immunity (2016) 44, 739-754, incorporated herein by reference.

[0144] STING agonists are immunomodulators responsible for regulating many host defense genes, including type I interferons and proinflammatory cytokines, after recognizing cyclic dinucleotides in the cytosol of cells. These signals can then stimulate the adaptive immune system through antigen cross-presentation and T cell priming, among other mechanisms (Barber GN. STING: infection, inflammation and cancer. Nat Rev Immunol. 2015;15(12):760-70). TLR and STING agonists can also promote antitumor immune responses in solid tumors and cancers treated with immunotherapy (Berger G, Marloye M, Lawler SE. Immunotherapy. Trends Mol Med.2019;25(5):412-427).

[0145] STING agonists were ADU-S100 and 2'3'-cG S A S MP. STING agonists include: cyclic dinucleotides and their analogs, e.g. [ka] .

[0146] STING agonists also include modified cyclic dinucleotides. In some embodiments, the modified cyclic dinucleotides may not be naturally occurring or may be chemically synthesized. In some embodiments, the modified cyclic dinucleotide has the formula: [ka] In some embodiments, R1 and R2 can each independently be 9-purine, 9-adenine, 9-guanine, 9-hypoxanthine, 9-xanthine, 9-uric acid, or 9-isoguanine, the structure of which is as follows: [ka] R1 and R2 may be the same or different. In some embodiments, the aryl groups described in US 2016 / 0287623 (herein incorporated by reference) are As described above, the compound may be provided in the form of a predominantly Rp.Rp or Rp.Sp stereoisomer, or a prodrug, or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound may be provided in the form of a predominantly Rp.Rp stereoisomer. In certain embodiments, the compound may be in the form of the following formula or a predominantly Rp.Rp stereoisomer thereof: [ka]

[0147] STING agonists can be represented by the formula: [ka] The compound may include:

[0148] STING agonists can be represented by the formula: [ka] The compound may include:

[0149] STING agonists can be represented by the formula: [ka] The compound may include:

[0150] STING agonists can be represented by the formula: [ka] The compound may include:

[0151] STING agonists can be represented by the formula: [ka] The compound may include:

[0152] STING agonists may be of the formula: [ka] The compound may include:

[0153] STING agonists can be represented by the formula: [ka] The compound may include:

[0154] STING agonists can be represented by the formula: [ka] The compound may include:

[0155] STING agonists can be represented by the formula: [ka] The compound may include:

[0156] STING agonists can be represented by the formula: [ka] The compound may include:

[0157] STING agonists can be represented by the formula: [ka] The compound may include:

[0158] STING agonists can be represented by the formula: [ka] The compound may include:

[0159] STING agonists can be represented by the formula: [ka] The compound may include:

[0160] STING agonists can be represented by the formula: [ka] The compound may include:

[0161] STING agonists can be represented by the formula: [ka] The compound may include:

[0162] STING agonists can be represented by the formula: [ka] The compound may include:

[0163] STING agonists can be represented by the formula: [ka] The compound may include:

[0164] STING agonists can be represented by the formula: [ka] The compound may include:

[0165] STING agonists can be represented by the formula: [ka] The compound may include:

[0166] STING agonists can be represented by the formula: [ka] The compound may include:

[0167] STING agonists can be represented by the formula: [ka] The compound may include:

[0168] STING agonists can be represented by the formula: [ka] The compound may include:

[0169] STING agonists may be of the formula: [ka] The compound may include:

[0170] STING agonists can be represented by the formula: [ka] The compound may include:

[0171] STING agonists can be represented by the formula: [ka] The compound may include:

[0172] STING agonists can be of the formula: [ka] The compound may include:

[0173] STING agonists can be represented by the formula: [ka] The compound may include:

[0174] STING agonists may be of the formula: [ka] The compound may include:

[0175] STING agonists can be of the formula: [ka] The compound may include:

[0176] STING agonists can be of the formula: [ka] The compound may include:

[0177] STING agonists can be represented by the formula: [ka] The compound may include:

[0178] STING agonists can be represented by the formula: [ka] The compound may include:

[0179] STING agonists can be of the formula: [ka] The compound may include:

[0180] STING agonists can be of the formula: [ka] The compound may include:

[0181] STING agonists can be represented by the formula: [ka] The compound may include:

[0182] STING agonists have the formula [ka] (In the formula, X, 1 ~X 3 , L, Q, Z, Y, n, and R 6 ~R 8 may include compounds as described in WO 2018 / 234805 (herein incorporated by reference).

[0183] STING agonists have the formula [ka] (In the formula, X, 1 ~X 3 , L, Q, Y, and R 6 ~R8 may include compounds as described in WO 2018 / 234807 (herein incorporated by reference).

[0184] STING agonists have the formula [ka] (In the formula, X, 1 ~X 3 , L, Q, Y, and R 6 ~R 11 may include compounds as described in WO 2018 / 234808 (herein incorporated by reference).

[0185] STING agonists include, for example, the compound DMXAA: [ka] Includes.

[0186] STING agonists include di-amidobenimidazoles, e.g., [ka] Includes.

[0187] Preferably, D is a cyclic dinucleotide, e.g. [ka] where Y is a nucleobase and X is O or S, as exemplified below. Nucleobases include naturally occurring purine and pyrimidine bases, as well as modified purine and pyrimidine bases, and other modified heterocyclic bases. Such modifications include methylated purines or pyrimidines, acylated purines or pyrimidines, etc. Modified nucleobases include, for example, deazapurines, N-1-methylguanosine, isoguanine, 2-aminopurine, 1,3-diaza-2-oxophenothiazine, 1,3-diaza-2-oxophenoxazine, 7-nitro-1,3-diaza-2-oxophenoxazine, 2,6-diaminopurine, purine, 6-thioguanine, hypoxanthine, 2-pyrimidinone, 2-pyridone, 4-thiouridine, imidazole-4-carboxamide, N-substituted 5-(carboxamido)uridine, such as 5-(N-benzylcarboxamido)-uridine, or 5-fluoro-deoxyuridine.

[0188] According to the definition of a payload moiety above, a "cyclic dinucleotide payload moiety" is It is a cyclic dinucleotide with its nucleophilic group (typically O) attached to a linker. For example, [ka] but, [ka] when the cyclic dinucleotide payload moiety is [ka] It could be.

[0189] In some embodiments, the payload D is: [ka] It is.

[0190] The compounds of formula (I) / (IA) are as follows: [ka] [ka] [ka] Includes.

[0191] Preferably, D is an imidazo[4,5-c]quinolin-4-amine, such as [ka] According to the definition of the payload moiety above, an "imidazo[4,5-c]quinolin-4-amine payload moiety" is an imidazo[4,5-c]quinolin-4-amine, excluding its nucleophilic group (typically O or N) that is attached to a linker. For example, [ka] but, [ka] when the imidazo[4,5-c]quinolin-4-amine payload moiety D' is [ka] For example, [ka] but, [ka] when the imidazo[4,5-c]quinolin-4-amine payload moiety D' is [ka] The compound of formula (I) / (IA) may be [ka] Includes.

[0192] Methods for the synthesis of trans-cyclooctene modified payloads are described in detail in WO 2018 / 187740, WO 2014 / 205126, WO 2015 / 139025, and WO 2017 / 044983, which are incorporated by reference herein.

[0193] The compounds may exist as stereoisomers in which asymmetric or chiral centers are present. The stereoisomers are either "R" or "S" depending on the configuration of the substituents around the chiral carbon atom. As used herein, the terms "R" and "S" are the configurations defined in the IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem., 1976, 45:13-30. The present disclosure contemplates various stereoisomers and mixtures thereof, which are expressly included within the scope of the present invention. Stereoisomers include enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. Individual stereoisomers of the compounds can be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers, or by preparation of racemic mixtures followed by resolution methods well known to those skilled in the art. Such resolution methods include: (1) Furniss, Hannaford, Smith, and Tatchell, “Vogel's Textbook of Practical Organic Chemistry”5 th (1) by combining the mixture of enantiomers with a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and optically liberating the optically pure product from the auxiliary, or (2) by directly separating the mixture of optical enantiomers on a chiral chromatography column, or (3) by fractional recrystallization methods, as described in U.S. Pat. No. 6,339,341, published by Longman Scientific & Technical in 1989, edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England.

[0194] It is to be understood that the compounds may have tautomeric forms and geometric isomers and that these also form an aspect of the present invention.

[0195] The present disclosure also includes isotopically labeled compounds, which are identical to the compounds described herein except that one or more atoms have been replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes suitable for inclusion in the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, including, but not limited to, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, respectively. 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, and 36 Cl. Deuterium (i.e. 2Substitution with heavier isotopes, such as 3H, may confer certain therapeutic advantages from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may be preferred in some circumstances. The compounds may incorporate positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron-emitting isotopes that may be incorporated into compounds of formula (I), (II-A), or (III-A) are: 11 C. 13 N, 15 O, and 18 F. Isotopically labeled compounds disclosed herein can generally be prepared by conventional techniques well known to those skilled in the art or by methods analogous to those illustrated in the accompanying examples, using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents.

[0196] B. Therapeutic Support Composition The therapeutic support composition includes a support. The support can be a biocompatible support composition, i.e., compatible with the subject's body. In some cases, the support is non-toxic to the subject and does not substantially react with tissues or biological compounds in the subject. For example, the support can be a hydrogel, among others. The support can be implanted in the subject's body and can support the binding agent (e.g., tetrazine-containing group) as well as the payload after the binding agent is conjugated. Exemplary supports include, but are not limited to, polymers, viscous or non-viscous liquid materials, gels, hydrogels, polysaccharide hydrogels, crosslinked polymer matrices, metals, ceramics, plastics, bone graft materials, alginates, cellulose, chitosan, hyaluronic acid, chondroitin sulfate, heparin, and the like. Supports also include particles, such as nanoparticles and microparticles.

[0197] The hydrogel may be a polysaccharide hydrogel, alginate, cellulose, hyaluronic acid, chitosan, chitosin, chitin, hyaluronic acid, chondroitin sulfate, heparin, etc. Other suitable sugar-based biomaterials include those described in Polymer Advanced Technology, 2014, 25, 448-460. Polymers that may be used as supports include, but are not limited to: polyphosphazenes, polyanhydrides, polyacetals, poly(orthoesters), polyphosphoesters, polycaprolactones, polyurethanes, polylactides, polycarbonates, polyamides, and polyethers, and blends / composites / copolymers thereof. Representative polyethers include, but are not limited to, poly(ethylene glycol) (PEG), poly(propylene glycol) (PPG), triblock Pluronic® (PEG), poly(propylene glycol) (PPG ... n -[PPG] m -[PEG]n), PEG diacrylate (PEGDA), and PEG dimethacrylate (PEGDMA). Supports can also include proteins and other poly(amino acids), such as collagen, gelatin, elastin and elastin-like polypeptides, albumin, fibrin, poly(γ-glutamic acid), poly(L-lysine), poly(L-glutamic acid), poly(aspartic acid), and the like.

[0198] In some embodiments, the support is a hydrogel. In some embodiments, the support is an alginate. In some embodiments, the support is chitin. In some embodiments, the support is hyaluronic acid (e.g., non-hydrogel hyaluronic acid substantially free of crosslinks). In some embodiments, the support is chitosin.

[0199] In certain embodiments, the support is a particle. The particle of the present disclosure may have a diameter of 2 cm or less, for example, a diameter of 1.5 cm or less, or 1 cm or less, or 0.5 cm or less. For example, the particle may be a nanoparticle or a microparticle. Nanoparticles include particles with an average dimension on the nanometer scale (e.g., 1000 nm or less). Microparticles are particles with an average dimension on the micrometer scale (e.g., 1000 μm or less). "Average" refers to the arithmetic mean. In some embodiments, the nanoparticles have a diameter in the range of 1 nm to 1 μm, e.g., 10 nm to 1 μm, or 25 nm to 1 μm, or 50 nm to 1 μm, or 75 nm to 1 μm, or 100 nm to 1 μm, or 150 nm to 1 μm, or 200 nm to 1 μm, or 250 nm to 1 μm, or 300 nm to 1 μm, or 350 nm to 1 μm, or 400 nm to 1 μm, or 450 nm to 1 μm, or 500 nm to 1 μm. In other embodiments, the microparticles have diameters in the range of 1 μm to 1 mm, e.g., 10 μm to 1 mm, or 25 μm to 1 mm, or 50 μm to 1 mm, or 75 μm to 1 mm, or 100 μm to 1 mm, or 150 μm to 1 mm, or 200 μm to 1 mm, or 250 μm to 1 mm, or 300 μm to 1 mm, or 350 μm to 1 mm, or 400 μm to 1 mm, or 450 μm to 1 mm, or 500 μm to 1 mm. In another embodiment, small particles as small as 10 to 100 nm in diameter can be assembled to form larger complexes (e.g., clusters or aggregates as small as 1 to 10 μm). The particles of the present disclosure may be substantially spherical, such that they have a substantially circular cross-section. Other particle shapes may be used, including, but not limited to, ellipsoids, cubes, cylinders, cones, needles, or other irregular shapes.

[0200] A "particle" can take the form of any manufactured material, molecule, cryptophane, virus, phage, etc. Particles can be composed of materials such as, but not limited to, metals, ceramics, plastics, glass, composites, polymers, hydrogels, etc. For example, particles can be made of inert materials such as alginate or iron oxide. In some cases, particles can be magnetic and can be formed from paramagnetic, superparamagnetic or ferromagnetic materials, or other materials that respond to magnetic fields. Furthermore, particles can be of any shape, for example, spheres, rods, asymmetric shapes. A particle, or a group of several particles in a complex, can be functionalized with a receptor that has a specific affinity to bind to or interact with a clinically relevant substrate. The receptor can be inherent to the particle itself. For example, the particle itself can be a virus or phage that has an inherent affinity for a particular substrate. Additionally or alternatively, particles can be functionalized by covalently binding to, or attaching or associating with, a receptor that specifically binds to or otherwise recognizes a clinically relevant substrate. The functionalized receptor may be an antibody, peptide, nucleic acid, phage, bacteria, virus, or any other molecule with a defined affinity for the target substrate. Examples of materials that can be used for the "particle" and / or "support" include polylactic acid, polyglycolic acid, PLGA polymers, alginates and alginate derivatives, gelatin, collagen, fibrin, hyaluronic acid, laminin-rich gels, agarose, natural and synthetic polysaccharides, polyamino acids, polypeptides, polyesters, polyanhydrides, polyphosphazines, poly(vinyl alcohol), poly(alkylene oxide), poly(allylamine) (PAM), poly(acrylate), modified styrene polymers, Pluronic® polyols, poloxamers, poly(uronic acid), poly(vinylpyrrolidone), and copolymers or graft copolymers of any of these. These examples are not intended to limit the concentration, crosslinking with another agent, method of administration, engineered degradation profile, and other characteristics well known to those skilled in the art.

[0201] The particle, or a group of several particles in the complex, may be functionalized with a targeting agent (e.g., a ligand or antibody) that specifically binds (or substantially specifically binds) to a target (e.g., a target receptor or cell surface target, e.g., a clinically relevant receptor or cell surface target (e.g., an antigen)). The targeting agent may be directly attached to the particle itself. The targeting agent may be an antibody, peptide, nucleic acid, phage, bacteria, virus, or any other molecule that has a specific affinity for a target receptor or cell surface target. In some cases, the receptor or cell surface target is PD-1, CTLA-4, HER2 / neu, HER1 / EGFR, VEGFR, BCR-ABL, SRC, JAK2, MAP2K, EML4-ALK, BRAFV600E, 4-1BB, GITR, GSK3β, or other cell receptor or cell surface target. Other compounds or molecules (e.g., fluorescent dyes or autofluorescent or luminescent markers) that may aid in the detection (e.g., in vivo detection) of the particles may be attached to the particles. Ligands and / or detectable labels may be attached directly to the particles or via bioorthogonal functional groups as described herein.

[0202] In certain embodiments, the support is a bone graft material, such as a bone graft replacement material. A bone graft replacement material is a biocompatible material that is similar in structure to bone. In some cases, the bone graft replacement material is bioresorbable, which allows the bone graft replacement material to dissolve or be absorbed in the body over time. The bone graft replacement material may be osteoconductive, which promotes the formation of blood vessels and new bone into the bone graft replacement material. In some cases, the bone graft replacement material is osteoinductive, which promotes the formation of new bone through active recruitment of mesenchymal stem cells from the surrounding tissue. For example, the bone graft replacement material may contain growth factors such as bone morphogenetic proteins. Bone graft replacement materials include, but are not limited to, hydroxyapatite, tricalcium phosphate, demineralized bone matrix, bovine collagen, calcium sulfate, calcium phosphate, cancellous bone chips, and the like, as well as combinations thereof.

[0203] The therapeutic support composition of the present disclosure includes a support and a first binding agent covalently bonded to the support. The binding agent may be attached to the surface of the support, for example, a solvent accessible surface of the support (e.g., a surface of the support in contact with the surrounding solvent). In some cases, the binding agent is directly attached to the support. For example, the binding agent may be covalently bonded to the surface of the support through a covalent bond, for example, an amide, amine, ester, carbamate, urea, thioether, thiocarbamate, thiocarbonate, thiourea, etc. In some cases, the binding agent is covalently bonded to the support through an amide bond. In other cases, the binding agent may be linked to the support through a linker. Any suitable linker may be used to link the binding agent to the support. Exemplary linkers may have 1-100 linking atoms, including ethylene-oxy groups, amines, esters, amides, carbamates, and ketone functional groups. For example, the linker may have 1-50 linking atoms, or 5-50 linking atoms, or 10-50 linking atoms. Representative examples of linkers include, but are not limited to, the following: [ka]

[0204] In certain embodiments, the therapeutic support composition comprises a support and a compound of the formula: [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'' , NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; and t is 0, 1, 3, or 4. The tetrazine-containing group is

[0205] In certain embodiments, the therapeutic support composition has the formula: [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O) is selected from the group consisting of NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R'', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 22 is a linker of 1 to 100 linking atoms, which may include ethylene-oxy groups, amines, esters, amides, carbamates, and ketone functional groups. For example, the linker can have 1 to 50 linking atoms, or 5 to 50 linking atoms, or 10 to 50 linking atoms.

[0206] In certain embodiments, the therapeutic support composition has the formula: [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'' , NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; and t is 0, 1, 3, or 4. has.

[0207] In certain embodiments, the therapeutic support composition has the formula: [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'' , NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R'', R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; and R''' at each occurrence is independently selected from aryl and alkyl). or a salt thereof. Includes.

[0208] In certain embodiments, the therapeutic support composition has the formula: [ka] Includes units of.

[0209] In some embodiments, the therapeutic support composition has the formula: [ka] Includes units of.

[0210] In some embodiments, the therapeutic support composition has the formula: [ka] Includes units of.

[0211] In some embodiments, the therapeutic support composition has formula (II): [ka] (In the formula, G 2 teeth, [ka] ;R 22 is a linker of 1 to 100 linking atoms; R 20 are as defined herein) The substituted hyaluronic acid includes substituted hyaluronic acids having the units:

[0212] In another embodiment, G 2 teeth, [ka] It is.

[0213] In yet another embodiment, G 2 teeth, [ka] ;R 20 is hydrogen or C 1~4 It is an alkyl.

[0214] The compound of formula (II) may be represented by formula (III): [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'' , NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R'', R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; and R''' at each occurrence is independently selected from aryl and alkyl). In another embodiment according to formula (III), R 20 is hydrogen or C 1~4 It is an alkyl.

[0215] In some embodiments, the therapeutic support composition has the formula: [ka] Includes units of.

[0216] Another therapeutic support composition is described in WO 2017 / 044983, WO 2015 / 139025A1, and WO 2014 / 205126A1, the entire contents of each of which are incorporated herein by reference in their entirety.

[0217] Hyaluronic acid derivatives include hyaluronic acid having multiple glucuronic acid units and tetrazine-containing groups linked or directly bonded to the glucuronic acid units of hyaluronic acid.Hyaluronic acid may also have multiple N-acetylglucosamine units.In certain embodiments, the N-acetylglucosamine units of hyaluronic acid are not linked or conjugated to the tetrazine-containing groups.

[0218] The tetrazine-containing group is linked or directly bonded via the carboxylic acid of the glucuronic acid unit. The tetrazine-containing group can be incorporated into about 0.1% to about 80% of hyaluronic acid, measured by the percentage of carboxylic acid linked or conjugated with the tetrazine-containing group, for example, about 1% to about 75%, about 5% to about 75%, about 10% to about 50%, or about 40% to about 75% of hyaluronic acid, measured by the percentage of carboxylic acid linked or conjugated with the tetrazine-containing group.

[0219] 3.Synthesis method The disclosed compounds can be better understood in connection with the following synthetic schemes and methods that illustrate means by which the compounds can be prepared.

[0220] Generally, the compounds of formula (IA) / (IB) / (II-A) / (III-A) can be prepared by reacting a suitably activated linker with a payload having a primary amine, a secondary amine, or a hydroxyl group, either before or after the linker is attached to the cyclooctene moiety. It should be understood that the reactive group on the linker (e.g., ester, carbonate, acyl chloride, carboxylic acid) can be located at any selected site on the linker group. Conversely, the linker can contain a nucleophilic amine or hydroxyl group, such as an aldehyde, ketone, ester, carbonate, carboxylic acid, or acyl chloride, that can react with a suitable group on the payload.

[0221] In certain embodiments, trans-cyclooctene activated for nucleophilic addition can be reacted with an appropriate payload (D / D) in the presence of a base as shown below. 1 ), or a linker L 4The payload can be reacted with a payload attached to -H to obtain a functionalized payload. The payload or linker can contain a primary amine, a secondary amine, or a hydroxyl group that reacts with the activated TCO. In certain embodiments, the leaving group (LG) is a chloro leaving group, a p-nitrophenol leaving group, or an N-hydroxysuccinimide leaving group. Exemplary bases used in this reaction include organic and inorganic bases, such as triethylamine, pyridine, sodium hydroxide, and sodium bicarbonate.

[0222] As shown in Scheme 1, trans-cyclooctene with an activated carbonate was converted to (D / D 1 )-L 4 -H to give intermediate 4, which can be further hydrolyzed to acid 5 or reacted under basic conditions to give amine G 1 -N(R 1c )H to give 7. G suitable for the method of Scheme 1 1 -N(R 1c )H, for example, HN(R 1c )CHR 1e CO2H, HN(R 1c )-C 1~6 Alkylene-COH, HN(R 1c )CHR 1e C(O)OC 1~4 Alkyl, and HN(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 Examples of such alkyl groups include alkyl. [ka]

[0223] As shown below in Scheme 2, trans-cyclooctene bearing an activated carbonate ester can be coupled with an amine-bearing payload (e.g., doxorubicin, abbreviated as doxo). Intermediate 4 can be hydrolyzed to an acid to provide a functionalized payload of the invention. [ka]

[0224] Scheme 3 shows another application of the above chemistry, in which the intermediate carbonate is reacted with the ornithine side chain of daptomycin, followed by the addition of the amino-containing group G under basic conditions. 1 -N(R 1c )H. [ka]

[0225] Scheme 4 shows the synthetic sequence for converting intermediate 10 to intermediate 11. Either 10 or 11 can be used to make linkers, protected linkers, or linkers attached to payloads using the general synthetic methods disclosed in WO2017 / 044983. The trimethylsilylethyl group can be removed at an appropriate point in the synthetic sequence to obtain the carboxylic acid. One skilled in the art will be able to modify the synthetic routes and protecting group strategies to achieve the compounds of the invention. [ka]

[0226] For example, Scheme 5 shows the conversion of 11 to a carboxylic acid intermediate, which can be further converted to payload-bearing products 13 and 14. [ka]

[0227] Other carboxylic acids that can be prepared using 11 are shown in Scheme 6. Payload D' in Schemes 5 and 6 can be prepared using payloads D or D. 1 The payload portion of either [ka]

[0228] Scheme 7 shows a general method for preparing TCO conjugates that contain amide substitutions on the TCO. [ka]

[0229] A synthetic strategy for preparing a representative STING agonist TCO conjugate is shown in Scheme 8. [ka]

[0230] A synthetic strategy for preparing a representative STING agonist TCO conjugate is shown in Scheme 9. [ka]

[0231] Scheme 10 shows a general method for conjugating a cyclic dinucleotide to trans-cyclooctene as shown in formula (I). The exemplified method is the conjugation of a carbonate nitrile in the presence of a base. The method proceeds by reacting a cyclic dinucleotide with a diphenyl substituted trans-cyclooctene to form a mono- or bis-substituted cyclic dinucleotide, depending on the amount of trans-cyclooctene. [ka]

[0232] Scheme 11 shows a general method for conjugating cyclic dinucleotides with trans-cyclooctene, where R 2 -C 1~6 Alkylene -COH, -CHR 1e CO2H, -C 1~6 Alkylene-C(O)OC 1~4 Alkyl, C(O)OC 1~4 Alkyl or -CHR 1e C(O)OC 1~4 alkyl, which is C(O)N(R 1c)-C 1~6 Alkylene-COH, C(O)OH, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 Alkyl, C(O)OC 1~4 Alkyl, or C(O)N(R 1c )CHR 1e C(O)OC 1~4 R of formula (I) is alkyl 1b The process of Scheme 11 proceeds similarly to that shown in Schemes 1-3, 7, and 9. As in Scheme 10, the process shown in Scheme 11 may be modified to provide bis-conjugated cyclic dinucleotides using an excess of trans-cyclooctene reagent. [ka]

[0233] Schemes 12 and 13 show representative synthetic methods for conjugating imidazo[4,5-c]quinolin-4-amine to trans-cyclooctene as shown in formula (I) following procedures similar to those in Schemes 10 and 11. [ka] [ka]

[0234] The disclosed compounds can be prepared in racemic form or as individual enantiomers or diastereomers by either stereospecific synthesis or resolution. The compounds can be resolved into their component enantiomers or diastereomers by standard techniques, for example, the formation of stereoisomeric pairs by salt formation with optically active bases, followed by fractional crystallization and regeneration of the free acid. The compounds can also be resolved by the formation of stereoisomeric esters or amides, followed by chromatographic separation and removal of the chiral auxiliary. Alternatively, the compounds can be resolved using a chiral HPLC column. Enantiomers can also be obtained from kinetic resolution of the corresponding ester racemates using lipase enzymes.

[0235] The compounds described herein may be in the form of a salt, e.g., a pharma- ceutically acceptable salt. The term "pharma- ceutically acceptable salt" includes salts of active compounds prepared with relatively non-toxic acids or bases, depending on the particular substituents present in the compounds described herein. The neutral form of the compound can be regenerated by contacting the salt with a base or acid, followed by isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise remains the same. For purposes of this disclosure, the salts are equivalent to the parent compound. Examples of pharma- ceutically acceptable salts are described in Berge et al., 1977, “Pharmaceutically Acceptable Salts." J. Pharm. Sci. Vol. 66, pp. 1-19.

[0236] For example, if the compound is anionic, or if a functional group which may be anionic (e.g., -COOH can be replaced with -COO - In the case where the cation has a substituent, a salt can be formed with a suitable cation. Examples of suitable inorganic cations include, but are not limited to, alkali metal ions (e.g., Na + and K +), alkaline earth cations (e.g., Ca 2+ and Mg 2+ ), as well as other cations. Examples of suitable organic cations include, but are not limited to, ammonium ion (i.e., NH + ) and substituted ammonium ions (e.g., NH3R1 + , NH2R2 + , NHR3 + , NR4 + Some examples of suitable substituted ammonium ions include those derived from: ethylamine, diethylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, and amino acids such as lysine and arginine.

[0237] When the compound is cationic, or a functional group which may be cationic (e.g., -NH2 may be -NH3 + In the case where the cation is an inorganic acid, it may be possible to form a salt with a suitable anion. Examples of suitable inorganic anions include, but are not limited to, those derived from the following inorganic acids: hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfurous acid, nitric acid, nitrous acid, phosphoric acid, and phosphorous acid.

[0238] Examples of suitable organic anions include, but are not limited to, those derived from the following organic acids: 2-acetoxybenzoic acid, acetic acid, ascorbic acid, aspartic acid, benzoic acid, camphorsulfonic acid, cinnamic acid, citric acid, edetic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, hydroxymaleic acid, hydroxynaphthalenecarboxylic acid, isethionic acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, methanesulfonic acid, mucic acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pantothenic acid, phenylacetic acid, phenylsulfonic acid, propionic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, tartaric acid, toluenesulfonic acid, and valeric acid. Examples of suitable polymeric organic anions include, but are not limited to, those derived from the following polymeric acids: tannic acid, carboxymethyl cellulose.

[0239] Unless otherwise specified, when a particular compound is referenced, its salt forms are also included.

[0240] It may be convenient or desirable to prepare, purify and / or handle active compounds in a chemically protected form. The term "chemically protected form" is used herein in its conventional chemical sense and refers to a compound in which one or more reactive functional groups are protected from undesired chemical reactions under specified conditions (e.g., pH, temperature, radiation, solvents, etc.). In practice, known chemical methods are used to reversibly render otherwise reactive functional groups unreactive under specified conditions. In a chemically protected form, one or more reactive functional groups are in the form of protected or protecting groups (also known as masked or blocking groups). By protecting reactive functional groups, reactions involving other unprotected reactive functional groups can be carried out without affecting the protected groups; protecting groups can usually be removed in a later step without substantially affecting the remainder of the molecule. For example, Protective See, for example, Ve Groups in Organic Synthesis (T. Green and P. Wuts; 3rd Edition; John Wiley and Sons, 1999). Unless otherwise specified, a reference to a particular compound also includes chemically protected forms thereof.

[0241] A wide variety of such "protecting," "blocking," or "masking" methods are widely used and well known in organic synthesis. For example, a compound having two unequal reactive functional groups (either of which may be reactive under specified conditions) may be derivatized to "protect" one of the functional groups, and therefore render it unreactive, under specified conditions; so protected, the compound may in effect be used as a reactant with only one reactive functional group. After the desired reaction (involving the other functional group) is complete, the protected group may be "deprotected" to return it to its original functionality.

[0242] Hydroxy groups can be protected as ethers (-OR) or esters (-OC(O)R), for example, as t-butyl ethers; benzyl, benzhydryl (diphenylmethyl), or trityl (triphenylmethyl) ethers; trimethylsilyl or t-butyldimethylsilyl ethers; or acetyl esters (-OC(O)CH, -OAc).

[0243] Aldehyde or ketone groups can be protected, for example, as acetals (RCH(OR)2) or ketals (R2C(OR)2) in which the carbonyl group (R2C=O) is converted to a diether (R2C(OR)2) by reaction with a primary alcohol, respectively, which is readily regenerated by hydrolysis using a large excess of water in the presence of acid.

[0244] Amine groups can be protected, for example, as amides (-NRC(O)R) or urethanes (-NRC(O)OR), e.g., as methylamides (-NHC(O)CH3); benzyloxyamides (-NHC(O)OCH2C6H5, -NH-Cbz); t-butoxyamides (-NHC(O)OC(CH3)3, -NH-Boc); 2-biphenyl-2-propoxyamides (-NHCO(O)C(CH3)2C6H4C6H5, -NH-Bpoc), 9-fluorenylmethoxyamides. (-NH-Fmoc), as 6-nitroveratryloxyamide (-NH-Nvoc), as 2-trimethylsilylethyloxyamide (-NH-Teoc), as 2,2,2-trichloroethyloxyamide (-NH-Troc), as allyloxyamide (-NH-Alloc), as 2(-phenylsulfonyl)ethyloxyamide (-NH-Psec); or, in appropriate cases (e.g. cyclic amines), as a nitroxide radical (>NO<<).

[0245] Carboxylic acid groups can be protected as esters, for example as alkyl esters (e.g., methyl esters; t-butyl esters); haloalkyl esters (e.g., haloalkyl esters); trialkylsilylalkyl esters; or arylalkyl esters (e.g., benzyl esters; nitrobenzyl esters); or as amides, for example as methylamides.

[0246] Thiol groups can be protected as thioethers (-SR), for example, as benzyl thioethers; acetamidomethyl ethers (-S-CH2NHC(O)CH3).

[0247] The compounds described herein can also be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications can be performed using known methods known in the art. These include those that increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism, and / or alter excretion rate. Examples of these modifications include, but are not limited to, the following: esterification with polyethylene glycol, derivatization with pivorate or fatty acid substituents, conversion to carbamates, hydroxylation of aromatic rings, and heteroatom substitution in aromatic rings.

[0248] In certain embodiments, the products may be further modified, for example, by manipulation of substituents. Such manipulations include, but are not limited to, reduction, oxidation, organometallic cross-coupling, alkylation, acylation, and hydrolysis reactions, which are well known to those skilled in the art. In some cases, the order of carrying out the foregoing reaction schemes may be varied to facilitate the reaction or to avoid unwanted reaction products.

[0249] 4. Preparation Another aspect of the present invention provides a pharmaceutical composition comprising: a) a compound of formula (II-A), or a pharma- ceutically acceptable salt thereof; b) one or more immunomodulatory agents, or a pharma- ceutically acceptable salt thereof; and c) a pharma- ceutically acceptable carrier.

[0250] Another aspect of the present invention provides a pharmaceutical composition comprising: a) a therapeutic support composition; b) one or more immunomodulatory agents, or pharma- ceutically acceptable salts thereof; and c) a pharma- ceutically acceptable carrier.

[0251] The compositions (e.g., the support composition, one or more immunomodulatory agents, and / or functionalized payload) can be provided in any suitable form, e.g., in the form of a pharma- ceutically acceptable formulation, and can be formulated for any suitable route of administration, e.g., oral, topical, or parenteral. When the compositions are provided as injectable solutions (e.g., in embodiments in which they are administered intravenously or directly to a tissue), the compositions can be provided as ready-to-use dosage forms or as reconstitutable storage-stable powders or liquids, which can contain pharma- ceutically acceptable carriers and excipients.

[0252] A "pharmaceutically acceptable excipient," "pharmaceutically acceptable diluent," "pharmaceutically acceptable carrier," or "pharmaceutically acceptable adjuvant" is an excipient, diluent, carrier, and / or adjuvant useful in preparing a pharmaceutical composition, which generally includes excipients, diluents, carriers, and adjuvants that are safe, non-toxic, and not biologically or otherwise harmful and acceptable for veterinary and / or human pharmaceutical use. As used herein, "pharmaceutically acceptable excipient, diluent, carrier, and / or adjuvant" includes one or more excipients, diluents, carriers, and adjuvants.

[0253] Methods for formulating the compositions can be readily adapted from those available. For example, the compositions can be provided as pharmaceutical formulations that include a therapeutically effective amount of the composition and a pharma- ceutically acceptable carrier (e.g., saline). The pharmaceutical formulations can optionally contain other additives (e.g., buffers, stabilizers, preservatives, etc.). In some embodiments, the formulations are suitable for administration to a mammal, e.g., suitable for administration to a human.

[0254] The compositions of the present disclosure can be prepared into various oral, parenteral and topical dosage forms. Oral formulations include tablets, pills, powders, dragees, capsules, liquids, lozenges, cachets, gels, syrups, slurries, suspensions, etc., suitable for ingestion by a subject. The compositions of the present disclosure can also be administered by injection, which can be intravenous, intramuscular, intradermal, subcutaneous, intraduodenal, or intraperitoneal injection. In some cases, the compositions described herein can be , by inhalation, for example, intranasally. In some cases, the compositions of the present disclosure can be administered transdermally. In some cases, the compositions can be administered via ocular, intravaginal, and intrarectal routes, including suppositories, insufflation, powders, and aerosol formulations (e.g., steroid inhalants, see Rohatagi, J. Clin. Pharmacol. 35:1187-1193, 1995; Tjwa, Ann. Allergy Asthma Immunol. 75:107-111, 1995). Thus, the present disclosure also provides pharmaceutical formulations comprising the compositions described herein and a pharma- ceutically acceptable carrier or excipient.

[0255] For preparing pharmaceutical formulations from the compositions of the present disclosure, pharma- ceutically acceptable carriers can be solid or liquid. Solid formulations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules. A solid carrier can be one or more substances that can also act as diluents, flavoring agents, binders, preservatives, tablet disintegrating agents, or encapsulating materials. Details regarding techniques for formulation and administration can be found, for example, in Remington's Pharmaceutical Sciences, Maack Publishing Co, Easton PA ("Remington's").

[0256] In some embodiments, the pharmaceutical composition of the invention is a vaccine comprising a compound of formula (IA), or a pharma- ceutically acceptable salt thereof, a pharma- ceutically acceptable carrier, and optionally an antigen. The antigen used in the immunogenic compositions provided herein may be provided in an effective amount (e.g., an amount effective for use in a therapeutic or prophylactic method). For example, the immunogenic compositions of the invention may be used to treat or prevent a disease or condition, such as infectious disease and cancer. Exemplary antigens include, but are not limited to, tumor antigens and infectious disease antigens. The antigen used in the immunogenic compositions provided herein is typically a macromolecule (e.g., a polypeptide, polysaccharide, polynucleotide) that is foreign to the host. An antigen may be any target epitope, molecule (including a biomolecule), molecular complex (including a molecular complex containing a biomolecule), intracellular assembly, cell or tissue against which it is desired to induce or enhance immune reactivity in a subject. Often, the term antigen may refer to a polypeptide antigen of interest. However, antigen as used herein may also refer to a recombinant construct (e.g., an expression construct) that encodes a polypeptide antigen of interest. In certain preferred embodiments, the antigen may be, may be derived from, or may be immunologically cross-reactive with, an infectious agent and / or epitope, biomolecule, cell or tissue associated with an infectious disease, cancer, autoimmune disease, allergy, asthma, or any other medical condition to which an antigen-specific immune response may be desirable or beneficial.

[0257] In certain embodiments, tumor or cancer antigens are used with the immunogenic compositions provided herein. In certain embodiments, the tumor antigen is a peptide-containing tumor antigen, such as a polypeptide tumor antigen or a glycoprotein tumor antigen. In certain embodiments, the tumor antigen is a saccharide-containing tumor antigen, such as a glycolipid tumor antigen or a ganglioside tumor antigen. In certain embodiments, the tumor antigen is a polynucleotide-containing tumor antigen that expresses a polypeptide-containing tumor antigen, such as an RNA vector construct or a DNA vector construct, such as a plasmid DNA. In certain embodiments, the tumor antigen is a live or dead or permeable whole cancer cell. Tumor antigens suitable for use with the immunogenic compositions provided herein include a wide variety of molecules, such as (a) polypeptide-containing tumor antigens, such as polypeptides (which can be, for example, 8-20 amino acids in length, although lengths outside this range are common), lipopolypeptides, and glycoproteins, (b) saccharide-containing tumor antigens, such as polysaccharides, mucins, gangliosides, glycolipids, and glycoproteins, and (c) polynucleotides that express antigenic polypeptides.

[0258] In certain embodiments, tumor antigens include, for example, (a) full-length molecules associated with cancer cells, (b) ) homologues and modified forms thereof, e.g., molecules containing deleted, added and / or substituted portions, and (c) fragments thereof. In certain embodiments, the tumor antigen is provided in recombinant form. In certain embodiments, the tumor antigen comprises, for example, a class I-restricted antigen recognized by CD8+ lymphocytes or a class II-restricted antigen recognized by CD4+ lymphocytes.

[0259] In certain embodiments, tumor antigens include, but are not limited to, (a) cancer / testis antigens, such as NYESO-1, SSX2, SCP1, and RAGE, BAGE, GAGE ​​and MAGE family polypeptides, such as GAGE-1, GAGE-2, MAGE-1, MAGE-2, MAGE-3, MAGE-4, MAGE-5, MAGE-6, and MAGE-12 (which can be used, for example, to address melanoma, lung, head and neck, NSCLC, breast, gastrointestinal, and bladder tumors); (b) mutant antigens, such as p53 (associated with various solid tumors, for example, colon, lung, head and neck cancer), p21 / Ras (associated with melanoma, pancreatic cancer, and colon cancer), CDK4 (associated with melanoma), MUM1 (associated with melanoma), caspase-8 (associated with head and neck cancer), CIA 0205 (e.g., associated with bladder cancer), HLA-A2-Rl 701, β-catenin (e.g., associated with melanoma), TCR (e.g., associated with T-cell non-Hodgkin's lymphoma), BCR-abl (e.g., associated with chronic myeloid leukemia), triosephosphate isomerase, KIA 0205, CDC-27, and LDLR-FUT; (c) overexpressed antigens, e.g., galectin 4 (e.g., associated with colon cancer), galectin 9 (e.g., associated with Hodgkin's disease), proteinase 3 (e.g., associated with chronic myeloid leukemia), WT 1 (e.g., associated with various leukemias), carbonic anhydrase (e.g., associated with renal cancer), aldolase A (e.g., associated with lung cancer), PRAME (e.g., associated with melanoma), HER-2 / neu (e.g., associated with breast, colon, lung and ovarian cancer), alpha-fetoprotein (e.g., associated with hepatocellular carcinoma), KSA (e.g., associated with colorectal cancer), gastrin (e.g., associated with pancreatic and gastric cancer), telomerase catalytic protein, MUC-1 (e.g., associated with breast and ovarian cancer), G-250 (e.g., associated with renal cell carcinoma), p53 (e.g., associated with breast, colon cancer), and carcinoembryonic antigen (e.g., associated with cancers of the digestive tract, such as breast, lung and colorectal cancer); (d) common antigens, e.g., melanoma melanocyte differentiation antigens, e.g., MART-1 / Melan A, gp 100, MC1R, melanocyte stimulating hormone receptor, tyrosinase, tyrosinase related protein-1 / TRP1 and tyrosinase related protein-2 / TRP2 (e.g., associated with melanoma), (e) prostate-related antigens, e.g., associated with prostate cancer, e.g., PAP, PSA, PSMA, PSHP1, PSM-P1, PSM-P2, (f) immunoglobulin idiotypes (e.g., associated with melanoma and B cell lymphoma), and (g) other tumor antigens, such as polypeptide and carbohydrate-containing antigens, including: (i) glycoproteins, e.g., sialyl Tn and sialyl Tn. (ii) lipopolypeptides (e.g., MUC-1 linked to a lipid moiety); (iii) polysaccharides (e.g., globo H synthetic hexasaccharide) linked to a carrier protein (e.g., KLH); and (iv) gangliosides, such as GM2, GM12, GD2, GD3 (e.g., associated with brain tumors, lung cancer, melanoma), also linked to a carrier protein (e.g., KLH).

[0260] In certain embodiments, tumor antigens include, but are not limited to, p15, Hom / MeI-40, H-Ras, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, Epstein-Barr virus antigens, EBNA, human papillomavirus (HPV) antigens including E6 and E7, hepatitis B and C virus antigens, human T-cell lymphotropic viral antigens, TSP-180, p185erbB2, p180erbB-3, c-met, nm-23H1, TAG-72-4, CA 19-9, CA 72-4. , CAM 17.1, NuMa, K-ras, p16, TAGE, PSCA, CT7, 43-9F, 5T4, 791 Tgp72, β-HCG, BCA225, BTAA, CA 125, CA 15-3 (CA 27.29\BCAA), CA 195, CA 242, CA-50, CAM43, CD68\KP1, C0-029, FGF-5, Ga733 (EpCAM), HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90 (Mac-2 binding protein\cyclophilin C-related protein), TAAL6, TAG72, TLP, TPS, etc.

[0261] 5. Treatment method Aspects of the present disclosure include methods of delivering a payload to a target site in a subject. In certain embodiments, the methods include selectively delivering a payload to a target site in a subject. Selective delivery of a payload includes delivering a payload to a target site (e.g., an organ or tissue, or a portion thereof) without targeting other sites in the subject that do not require administration of the payload (e.g., other organs or tissues, or portions thereof). Selective delivery of a payload can be achieved using the support compositions and functionalized payloads described herein.

[0262] In some cases, the support composition of the present disclosure may be localized at a desired target site in a subject. For example, the method of the present disclosure may include administering a support composition described herein to a subject. The support composition may be administered to a subject at a desired target site in the subject. In some cases, the support composition may be implanted in a subject at a desired target site in the subject. In some embodiments, the support composition may be conjugated to a targeting agent described herein, and the method may include administering (e.g., systemically administering) the support composition to a subject. In these embodiments, the support composition conjugated to a targeting agent may be localized at a desired target site in a subject by specific binding of the targeting agent to its target (e.g., antibody-antigen interaction, etc.), or may be localized on the surface of the desired target (e.g., cell surface) by specific binding of the targeting agent to its target (e.g., antibody-antigen interaction, etc.).

[0263] As described herein, selective binding between orthogonal binding partners (e.g., between a tetrazine binding agent of the support composition and a complementary trans-cyclooctene binding agent of the functionalized payload) can occur. As previously described, upon localized administration of the support composition to a desired site in a subject, selective binding between the binding agent of the support composition and the complementary binding agent of the functionalized payload will localize the payload to the desired target site. Thus, in certain embodiments, the method includes administering a functionalized payload to a subject such that the functionalized payload binds to the support composition to form a support complex. For example, the functionalized payload can be administered systemically to a subject. Upon administration of the functionalized payload to a subject, contact occurs between the binding agent of the support composition and the complementary binding agent of the functionalized payload, such that the binding agent and its complementary binding agent bind to each other to form a support complex, thereby allowing the payload to be selectively delivered to a target site in the subject. In some embodiments, selective delivery of the functionalized payload achieves a higher concentration of the payload at the target site than at other locations in the subject (eg, non-target sites in the subject).

[0264] Indications for this approach include cancer, both hematologic and solid, infections, wound healing, stenosis, ischemia, revascularization, myocardial infarction, arrhythmias, vascular occlusion (thrombus due to anticoagulants), inflammation due to antiproliferative drugs, corticosteroids and derivatives, and / or NSAIDs, autoimmune disorders, grafts, macular degeneration, rheumatoid arthritis, osteoarthritis, coatings on implants, pastes, waxes, periprosthetic infections due to polymethylmethacrylate (PMMA) constructs, etc. In certain embodiments, the present approach can be used to treat and / or diagnose soft tissue sarcomas: rhabdomyosarcoma, fibrosarcoma, Ewing's sarcoma, and all subtypes of soft tissue sarcoma and osteosarcoma. The present compositions can be used to treat and / or diagnose pigmented villonodular synovitis.

[0265] The compositions of the present disclosure are useful for treating and / or diagnosing a condition or disease in a subject that is amenable to treatment or diagnosis by administration of a payload (e.g., a parent drug (i.e., the drug prior to conjugation to the composition)). "Treatment" means that at least an improvement in symptoms associated with the condition suffered by the subject is achieved, where improvement is used broadly and refers to at least a decrease in the magnitude of a parameter (e.g., a symptom) associated with the condition being treated. Thus, treatment also includes a condition in which the pathological condition, or at least a symptom associated therewith, is completely inhibited, e.g., prevented from occurring, or arrested, e.g., terminated, such that the subject is no longer afflicted by the condition, or at least a symptom characterizing the condition. Treatment may include inhibition, i.e., arrest of the occurrence or further occurrence of clinical symptoms, e.g., alleviation or complete inhibition of active disease. Treatment may include remission, i.e., causing regression of clinical symptoms. For example, in the context of cancer, the term "treating" includes any or all of the following: reducing the growth of solid tumors, inhibiting the replication of cancer cells, reducing overall tumor burden, increasing survival time, and ameliorating one or more symptoms associated with cancer.

[0266] The subject to be treated is one who needs a therapy, and the subject to be treated is one who is suitable for treatment with the parent drug. Thus, a variety of subjects are considered suitable for treatment with the compositions disclosed herein. In general, such subjects are "mammals", including humans. Other subjects include pets (e.g., dogs and cats), livestock (e.g., cows, pigs, goats, horses, etc.), rodents (e.g., mice, guinea pigs, and rats, e.g., as animal models of disease), and non-human primates (e.g., chimpanzees and monkeys).

[0267] The functionalized payload, therapeutic support composition, and method can be used for the treatment, prevention, and / or diagnosis of any target disease. Indications for this approach include cancer, both hematological and solid, infectious diseases, wound healing, stenosis, ischemia, revascularization, myocardial infarction, arrhythmias, vascular occlusion (thrombus due to anticoagulants), inflammation due to antiproliferative drugs, corticosteroids and derivatives, and / or NSAIDs, autoimmune disorders, grafts, macular degeneration, rheumatoid arthritis, osteoarthritis, coatings of implants, pastes, waxes, periprosthetic infections due to polymethylmethacrylate (PMMA) constructs, etc. In certain embodiments, the functionalized payload, therapeutic support composition, and method can be used for the treatment, prevention, and / or diagnosis of soft tissue sarcomas: rhabdomyosarcoma, fibrosarcoma, Ewing's sarcoma, and all subtypes of soft tissue sarcoma and osteosarcoma. The composition can be used for the treatment and / or diagnosis of pigmented villonodular synovitis.

[0268] In certain embodiments, the functionalized payload, therapeutic support composition, another therapeutic agent, one or more immunomodulatory agents, and methods can be used to treat, prevent, and / or diagnose solid tumors, including, but not limited to, melanoma (e.g., unresectable, metastatic melanoma), kidney cancer (e.g., renal cell carcinoma), prostate cancer (e.g., metastatic castration-resistant prostate cancer), ovarian cancer (e.g., epithelial ovarian cancer, such as metastatic epithelial ovarian cancer), breast cancer (e.g., triple-negative breast cancer), glioblastoma (e.g., glioblastoma multiforme), and lung cancer (e.g., non-small cell lung cancer), soft tissue sarcoma, fibrosarcoma, osteosarcoma, pancreatic cancer, among others. The disclosed approach is suitable as an adjuvant / neoadjuvant system. For example, during a biopsy, the particles disclosed herein can be placed, and once the test results are back, the physician can deliver the appropriate cocktail to the desired site of the body. This can minimize the size of the tumor, especially for tumors that can be surgically removed. Next, at the end of the operation, The physician can place more particles around the resection cavity and treat the patient with additional therapeutic agents (e.g., chemotherapy agents in the disclosed approach) to minimize the risk of any cancer cells that may have been missed at the resection margins.

[0269] In certain embodiments, the disclosed methods allow for the placement of the particles disclosed herein at the time of biopsy. Once the results are back, the physician can deliver immunomodulatory agents, such as TLR agonists, STING agonists, chemokines (agents that attract cancer cells and / or immune cells) and adjuvants to boost the immune system with fewer side effects, as well as chemotherapy agents combined with immunotherapeutic agents, through the biopsy site. This combination approach can benefit the patient. While the chemotherapy agents treat the solid tumor or specific site, the enhanced response of the immunotherapy can help distant metastatic sites. For example, in certain embodiments, the disclosed compositions and methods can be used with or in conjunction with anthracyclines, taxanes, gemcitabine, and other agents to enhance the effect of one or more immunomodulatory agents, such as ipilimumab, nivolumab, pembrolizumab, avelumab (alias: MSB0010718C; Pfizer).

[0270] The disclosed compounds and compositions can be used in methods of treatment. The disclosed methods of treatment can be used to treat bacterial infections. The disclosed methods of treatment can be used to treat or prevent MRSA infections. The disclosed methods of treatment can be used to treat cancer. The disclosed methods of treatment can be used to treat pigmented villonodular synovitis. The disclosed methods of treatment can be used to treat diseases or disorders related to inflammation. The disclosed methods of treatment can be used to treat arthritis.

[0271] Bacterial infection The disclosed method can be used to treat or prevent bacterial infections. Bacteria are not necessarily harmful and may be beneficial in some cases, but bacteria may also cause infections. Bacterial infections can affect multiple organs and body tissues, including but not limited to skin, mucous membranes, blood, lungs, kidneys, urinary tract, eyes, heart, intestines, meninges, airways, genitals, stomach, bone, connective tissue, and tissues surrounding organs. Bacterial infections can affect more than one organ or body tissue. Bacterial infections can be systemic. Bacterial infections can be asymptomatic. Bacterial infections can cause a variety of symptoms, including but not limited to fever, inflammation, non-healing wounds, oozing wounds, skin rash, skin erythema, abscesses, enlarged lymph nodes, nausea, diarrhea, headache, earache, sore throat, fatigue, low blood pressure, hyperventilation syndrome, weak and rapid pulse, local or generalized pain, and muscle pain. Bacterial infections can cause death. Subjects with comorbidities or compromised immune systems may be more susceptible to bacterial infections. Bacterial infections may occur at the surgical site. Bacterial infections may be associated with catheter placement.

[0272] Diagnosis of bacterial infection may include, but is not limited to, symptomatic diagnosis, microbial culture, microscopy, biochemical testing, PCR diagnosis, and metagenomic sequencing. Microbiological testing may include sample collection, microbial culture, identification, and antibiotic susceptibility testing. Diagnosis may include Gram staining of bacterial cultures. Diagnosis may include coagulase testing of bacterial cultures. Diagnosis may include catalase testing of bacterial cultures. Diagnosis may include blood tests. Blood tests may include, but are not limited to, complete blood count, measurement of C-reactive protein, measurement of procalcitonin, and measurement of rapid plasma reagin. Diagnosis may include ELISA. Diagnosis may include PCR. To identify MRSA, a rapid latex agglutination test detecting PBP2a protein may be performed. Samples may be grown on agar plates. Samples may be grown in nutrient broth. Growth conditions may include the ability of the growing bacteria to be resistant to the infection. Various factors may be included to determine the type (e.g., type of growth medium, nutrients, selective compounds, antibiotics, temperature, pH level, oxygen level). Determination of the bacteria growing on the agar plate or in the nutrient broth may determine the bacteria causing the subject's infection. A disk containing an antibiotic compound may be placed on the agar plate. The antibiotic compound may kill the bacteria growing on the plate. A larger zone of dead bacteria around the disk (zone of inhibition) may indicate greater antibiotic effectiveness.

[0273] A sample for diagnosing a bacterial infection may be obtained from a subject in need of treatment. The sample for testing may be from the site of the infection. The sample for testing may be obtained from the subject by swabbing the skin, throat, or nasal passages. The sample for testing may be obtained from the subject by collecting pus or fluid from a wound, abscess, or other skin infection. The sample for testing may be obtained from the subject by collecting a bodily fluid. The bodily fluid may include blood, saliva, urine, and / or other bodily fluids. Multiple samples may be taken from the subject. Multiple samples may be taken from around the prosthesis or medical device.

[0274] Bacterial infections may be treated using the compounds and compositions disclosed herein. Bacterial infections that may be treated by the compounds and compositions disclosed herein include, but are not limited to, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus (MRSA), Methicillin-susceptible Staphylococcus aureus (MSSA), Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Salmonella, Neisseria, Bacillus, Brucella, Nocardia, Listeria monocytogenes, Lactobacillus plantarum, Lactococcus lactis, Lactobacillus casei ... lactis, Francisella, Legionella, Yersinia pestis, Pseudomonas aeruginosa, Burkholderia cenocepacia, Mycobacterium avium, vancomycin-resistant Enterococci (VRE), and vancomycin-resistant Staphylococcus aureus (VRSA). The bacterial infection to be treated may be resistant to one or many antibiotics. The bacterial infection to be treated herein may be caused by gram-positive bacteria. The bacterial infection to be treated herein may be caused by vancomycin-resistant gram-positive bacteria. The bacterial infection to be treated herein may be caused by multi-drug resistant gram-positive bacteria.

[0275] i. MRSA infection The disclosed methods can be used to treat MRSA. MRSA is any strain of Staphylococcus aureus that has developed multiple resistance to β-lactam antibiotics, including penicillins (methicillin, dicloxacillin, nafcillin, oxacillin, etc.), and cephalosporins. MRSA evolved into at least five different strains of S. aureus from horizontal transfer of the mecA gene. MRSA infections can rapidly cause severe, life-threatening internal infections, including, but not limited to, sepsis, endocarditis, MRSA pneumonia, bone infections, and implant infections. MRSA can cause skin infections. MRSA skin infections can cause furuncles or abscesses. MRSA can cause systemic or internal infections. Some MRSA infections are untreatable with currently available antibiotics, commonly resulting in severe, debilitating infections or death. MRSA infections can also occur in subjects who have been hospitalized, known as healthcare-associated MRSA (HA-MRSA). MRSA infections can be spread by skin-to-skin contact, known as community-acquired MRSA (CA-MRSA). Cases of MRSA are increasing in livestock animals. A variant of MRSA, CC398, has emerged in animals and is found in intensively farmed production animals (e.g., pigs, cattle, and poultry), where MRSA can be transmitted to humans as LA-MRSA (livestock-associated MRSA).

[0276] Strains of MRSA to be treated with the compounds and compositions disclosed herein include, but are not limited to, the following: CBD-635, ST250 MRSA-1, ST2470-MRSA-I, ST239-MRSA-III, ST5-MRSA-II, ST5-MRSA-IV, ST239-MRSA-III, EMRSA15, EMRSA16, MRSA252, ST5:USA100, EMRSA 1, ST8:USA300, ST1:USA400, ST8:USA500, ST59:USA1000, USA1100, USA600, USA800, USA300, ST30, ST93, ST80, ST59, CC22, CC8, CC425, and CC398.

[0277] ii. Catheter-related bloodstream infection The disclosed method can be used to treat catheter-related bloodstream infection. Catheter-related bloodstream infection (CRBSI) is defined as the presence of bacteremia originating from an intravenous catheter. CRBSI is frequent, can be fatal, and can be a common cause of hospital-acquired bacteremia. Intravenous catheters are integral to modern technology practices and are inserted into critically ill patients for administration of fluids, blood products, medications, nutritional solutions, and hemodynamic monitoring. Central venous catheters (CVCs) may present a higher risk of device-related infection than any other type of medical device and can be a major cause of morbidity and mortality. They can be a cause of bacteremia and sepsis in hospitalized patients. CRBSI can be associated with CVCs.

[0278] The disclosed method can be used to deliver molecular payloads to implanted biomaterials (e.g., polymers or hydrogels substituted with bioorthogonal groups). The materials can be implanted in the desired location of the body during any local operation (e.g., surgical implant or indwelling device insertion ("local injection")), even when a specific pathogen or problem is not yet known. For example, a suitably modified polymer or hydrogel, e.g., tetrazine-modified hyaluronic acid (HAT), can be used to coat catheter materials or other implanted medical devices using known procedures for coating plastic materials with hyaluronic acid. The coating procedure can be optimized on small pieces of polyurethane (PU) or polyvinyl chloride (PVC) tubing. PU or PVC tubing can be treated with 3-aminopropyltriethoxysilane in distilled water to incorporate amine groups for covalent functionalization with hyaluronic acid (HA). A substratum of HAT or unmodified HA can then be attached to the surface using carbodiimide chemistry conditions detailed in the literature. Further layers of HAT or HA can be deposited by manually repeating the dip-coating procedure using similar carbodiimide chemistry conditions until a total of 10 more layers have been applied. The final coated tubes can be characterized by scanning electron microscopy to examine surface morphology, by confocal microscopy to determine coating thickness, and by contact angle measurements to assess surface hydrophilicity.

[0279] After implantation of the biomaterial-coated device, an inactive prodrug, made by modifying the drug with a reactive partner, is injected into the bloodstream wherever needed ("systemic exposure"). The inactive prodrugs spread throughout the body, but when they reach the vicinity of the biomaterial, whether in the form of a coating or gel, the prodrugs rapidly attach to the biomaterial ("trap"), thereby concentrating the prodrug at the desired site. Finally, the active drug is spontaneously released from the biomaterial to perform its function ("release"). This provides a system with temporal control of systemic drug delivery, effectively turning a systemic drug into a localized agent (Figure 8).

[0280] The limited systemic effect of the prodrug prevents problems with disrupting the body's natural microbiome or developing infectious diseases, such as drug-resistant bacteria. The highest therapeutic doses can be administered, which can increase the therapeutic index of the drug and reduce the chance of bacteria at the site of infection developing resistance. By coating the surface of the CVC or other implanted device with gel, the drug can accumulate deep in tissues that cannot be reached by systemic drugs at normal doses.

[0281] The disclosed method may achieve "reloading" with a prodrug to ensure improved local release and efficacy. This will lead to better utilization of the antibacterial agent and reduced emergence of resistance. If the bacterial or fungal infection proves resistant to the first prodrug, the second prodrug can be "captured and released" by the already implanted gel or coating device. Standard techniques require removal and placement of an implant to achieve a similar result. The disclosed biodegradable coating does not require additional invasive procedures to implant or remove it.

[0282] b. Cancer The disclosed method can be used to treat or prevent cancer. Cancer is a group of related diseases that may involve sustained growth signaling, evasion of growth inhibitors, resistance to cell death, achieving replicative immortality, induction of angiogenesis, and activation of invasion and metastasis. The disclosed method can enhance or induce an immune response against cancer in a subject. The immune response can result in an increase in one or more of leukocytes, lymphocytes, monocytes, and eosinophils.

[0283] Cancers that may be treated by the disclosed methods include, but are not limited to, astrocytoma, adrenocortical carcinoma, appendix cancer, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumor, brain stem cancer, brain stem glioma, breast cancer, cervical cancer, colon cancer, colorectal cancer, cutaneous T-cell lymphoma, diffuse pontine glioma, ductal carcinoma, endometrial cancer, ependymoma, Ewing's sarcoma, esophageal cancer, eye cancer, fibrosarcoma, bladder cancer, gastric cancer, gastrointestinal cancer, germ cell tumor, glioma, hepatocellular carcinoma, histiocytosis, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, Kaposi's sarcoma, renal cancer, laryngeal cancer, cancer), leukemia, liver cancer, lung cancer, lymphoma, macroglobulinemia, melanoma, mesothelioma, oral cancer, multiple myeloma, nasopharyngeal cancer, neuroblastoma, non-Hodgkin's lymphoma, osteosarcoma, ovarian cancer, pancreatic cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary cancer, prostate cancer, rectal cancer, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, skin cancer, small cell lung cancer, small intestine cancer, soft tissue cancer, soft tissue sarcoma, solid tumor, squamous cell carcinoma, stomach cancer, T-cell lymphoma, testicular cancer, pharyngeal cancer, thymoma, thyroid cancer, trophoblastic tumor, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, and Wilms' tumor.

[0284] In some embodiments, the cancer that may be treated by the disclosed methods is melanoma, renal cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a soft tissue cancer. In some embodiments, the cancer is a fibrosarcoma. In some embodiments, the cancer is diffuse pontine glioma.

[0285] Without wishing to be bound by any particular theory, certain anti-cancer Local release of drugs can cause or contribute to immunogenic cell death (ICD). For example, certain anticancer drugs (e.g., anthracyclines, cyclophosphamide, oxaliplatin) have been reported to induce ICD. Kroemer et al. Annu. Rev. Immunol. 2013(31), 51-72. Immunogenic apoptosis of cancer cells can induce effective antitumor immune responses by activating dendritic cells (DCs) and consequently specific T cell responses. ICD is characterized by the secretion of damage-associated molecular patterns (DAMPs). During ICD, three important DAMPs are exposed on the cell surface. One of the DAMP molecules, calreticulin (CRT), is normally located within the lumen of the endoplasmic reticulum (ER), but is translocated to the surface of dead cells after induction of immunogenic apoptosis, where it functions as an "eat me" signal for professional phagocytes. Other important surface-exposed DAMPs are the heat shock proteins (HSPs), namely HSP70 and HSP90, which also translocate to the plasma membrane under stress conditions. On the cell surface, they have immunostimulatory effects depending on their interaction with several antigen-presenting cell (APC) surface receptors, such as CD91 and CD40, and also promote the cross-presentation of antigens derived from tumor cells on MHC class I molecules, which subsequently leads to CD8+ T cell responses. Other important DAMPs specific to ICD are secreted amphoterin (HMGB1) and ATP. HMGB1 is considered a late apoptotic marker, and its release into the extracellular space appears to be necessary for optimal release and presentation of tumor antigens to dendritic cells. It binds to several pattern recognition receptors (PRRs), such as Toll-like receptors (TLR) 2 and 4, expressed on APCs. A recently recognized DAMP released during immunogenic cell death is ATP, which, upon secretion, acts as a "find-me" signal for monocytes, inducing their recruitment to sites of apoptosis. Kroemer et.al. Curr. Op. Immunol. 2008(20), 504-511.

[0286] Thus, localized delivery of ICD-inducing factors using the compounds and methods of the present invention may be beneficial in combination with one or more immunomodulatory agents.

[0287] In one aspect, the present invention provides a method of treating cancer, comprising the steps of: a) administering a therapeutically effective amount of a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt or composition thereof, to a subject in need thereof; and b) locally administering to a first tumor of the subject a therapeutic support composition described herein; wherein the subject has a second tumor, and wherein administering a) and administering b) inhibits the growth of the second tumor.

[0288] Another aspect provides a method of enhancing or eliciting an immune response against a second tumor in a subject, comprising the steps of: a) administering to the subject a therapeutically effective amount of a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt or composition thereof; and b) locally administering to a first tumor in the subject a therapeutic support composition described herein; wherein administering a) and administering b) enhances or elicits an immune response against the second tumor.

[0289] In another aspect, the present invention provides a method of inhibiting tumor metastasis in a subject at risk of tumor metastasis, comprising the steps of: a) administering to the subject a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof; and b) locally administering to a first tumor of the subject a therapeutic support composition; wherein the compound of Formula (II-A) or (III-A) and the therapeutic support composition are as defined herein.

[0290] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A), or a pharma- ceutically acceptable salt, or composition thereof; and b) a therapeutic support composition, for use in a method of inhibiting the growth of a second tumor in a subject, wherein the therapeutic support composition is administered locally to a first tumor in the subject and delivers the compound of formula (II-A) or (III-A), or The pharma- ceutically acceptable salt, or composition, is administered to a subject.

[0291] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, for use in a method of enhancing or inducing an immune response against a second tumor in a subject, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0292] In another aspect, the present invention provides a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, for use in a method of inhibiting tumor metastasis in a subject at risk of tumor metastasis, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0293] In another aspect, the present invention provides the use of a pharmaceutical combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, in the manufacture of a medicament for inhibiting the growth of a second tumor, wherein the therapeutic support composition is administered locally to a first tumor in a subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0294] In another aspect, the present invention provides the use of a combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof; and b) a therapeutic support composition, in the manufacture of a medicament for enhancing or inducing an immune response against a second tumor, wherein the therapeutic support composition is administered locally to a first tumor in a subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof, is administered to the subject.

[0295] In another aspect, the present invention provides the use of a combination comprising: a) a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and b) a therapeutic support composition, in the manufacture of a medicament for inhibiting tumor metastasis in a subject at risk of tumor metastasis, wherein the therapeutic support composition is administered locally to a first tumor in the subject, and the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition is administered to the subject.

[0296] The method of inhibiting tumor metastasis in a subject at risk of tumor metastasis may further include identifying and / or selecting a subject at risk of tumor metastasis. Subjects at risk of tumor metastasis can be identified from a tumor biopsy to assess the pathological status of the tumor by serum and / or tissue biomarkers and / or imaging techniques.

[0297] In the methods and uses disclosed herein, the second tumor may or may not be present in the subject at the time of administration of the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and the therapeutic support composition. In the methods and uses disclosed herein, the compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; and the therapeutic support composition may inhibit (i.e., prevent) the formation or development of the second tumor. In some embodiments, the therapeutic support composition is not administered locally to the second tumor. The methods of treating cancer or methods of enhancing or inducing an immune response and the disclosed pharmaceutical combinations may further be combined with the use of an immunomodulatory agent. Alternatively, the optional immunomodulatory agent may be omitted.

[0298] Without being bound by a particular theory, the methods and uses disclosed herein can inhibit metastasis or the formation or growth of secondary tumors by inducing or enhancing an immune response against the primary tumor (localized therapeutic support composition) and / or the secondary tumor (non-localized therapeutic support composition). The immune response can be an increase or decrease in one or more of innate and adaptive immune cells. For example, the immune response can be an increase or decrease in one or more of leukocytes, lymphocytes, monocytes, eosinophils, and antibodies. Further, for example, the immune response can be an increase in CD3, CD4, CD8, and / or PD-1 positive tumor infiltrating lymphocytes in the first tumor and / or the second tumor. The immune response can also be a decrease in regulatory T cells in the first tumor and / or the second tumor.

[0299] Without being bound to a particular theory, treatment of mouse breast and fibrosarcoma tumors with doxorubicin results in IFN-g-producing CD8+ T cell proliferation and their recruitment to tumor A is observed similar to that observed with radiation therapy, and the same types of cytotoxic compounds (e.g., anthracyclines, cyclophosphamide, and oxaliplatin) also activate immunogenic cell death pathways, resulting in cell surface expression of calreticulin followed by release of ATP, HMGB1, and HSPs, leading to DC-mediated cross-presentation of tumor antigens to CD8+ T cells. Confirmatory in vitro evidence shows that exposure of cancer cells to 5-fluorouracil or doxorubicin stimulates HSP release and promotes phagocytosis of cellular debris by DCs, thereby promoting cross-presentation to CD8+ T cells. Similarly, when doxorubicin-treated cancer cells are injected into allograft mice, DCs phagocytose cellular debris and generate tumor-specific CD8+ T cell antitumor immune responses (Medler TR, et. al. Trends Cancer. 2015;1(1):66-75.).

[0300] Inhibition of metastasis or secondary tumors not treated with therapeutic support composition may cause cell death in treated tumors. Cell death may result in release of stress molecules and antigens to the tumor microenvironment. These antigens may be presented by antigen-presenting cells to cytotoxic T cells, which may induce local and systemic immune responses against cells with similar antigens at the location of the second tumor. This treatment may recruit macrophages, NK cells, and cytotoxic T cells to the secondary tumor, which may result in a general increase in tumor-infiltrating lymphocytes and subsequent antitumor immune responses in the secondary tumor.

[0301] The method may be used to inhibit metastasis of solid malignant tumors in subjects at risk for tumor metastasis. Subjects at risk for tumor metastasis include those with stage IV (metastatic disease) including multiple tumors, or stage II-III (local spread). Subjects at risk for tumor metastasis also include those with high-grade solid tumors and those exhibiting tissue and / or serum biomarkers indicative of metastasis. Tumors classified as either grade 3 or "high grade" have poor cellular tissue differentiation and spread faster than grade 1 and 2 tumors. Biomarkers of metastasis include, but are not limited to, the following: CCR7, E-cadherin, CXCR4, VEGF, VEGFR, E-cadherin, EpCAM, VCAM, integrin-α10, N-cadherin, vimentin, and fibronectin. Additional biomarkers include the following, as described in Brinton et al., Cancer Genomics & Proteomics (2012) 9:345-356, incorporated herein by reference: AGR2, AGR3, alpha-enolase, CA125, CRP, SAA, IL6, IL8, CacyBP, CCR7, E-cadherin, CXCR4, CYFRA21-1, EGFR, EMP2, EphA2, galectin-1, GDF15, H2K18ac, H3K4me2, H3K9me2, HE4, HER2-neu, HSP27, HSP60, IGFBP2, IGFBP3, IGFBP7, IL6, IL6sR, ILK, integrin alpha v β6, LCN2, MSLN, Muc-1, PDX6, Pre These include cutin, SAA, SPARC, TFF3, TGF-β1, TGM2, TGM4, triosephosphate isomerase, USP9X, VCAM-1, VEGF-C, VEGF-D, and VVEGFR-3.

[0302] The biomarkers can be protein biomarkers, which can indicate risk of tumor metastasis by either increased or decreased protein expression compared to standard samples from non-metastatic or non-cancerous controls.

[0303] In some embodiments, the first tumor cells are isolated from a first tumor in a subject at risk for metastasis, hi other embodiments, the first tumor cells are present in tissue surrounding the first tumor, present in tumor cell-platelet aggregates, present in the systemic circulation of the subject, and / or present at the location of a second tumor in the subject.

[0304] In certain embodiments, the functionalized payloads, therapeutic support compositions, and methods may be used to treat, prevent, and / or diagnose solid tumors, including, but not limited to, melanoma (e.g., unresectable, metastatic melanoma), kidney cancer (e.g., renal cell carcinoma), prostate cancer (e.g., metastatic castration-resistant prostate cancer), ovarian cancer (e.g., epithelial ovarian cancer, such as metastatic epithelial ovarian cancer), breast cancer (e.g., triple-negative breast cancer), glioblastoma (e.g., glioblastoma multiforme), and lung cancer (e.g., non-small cell lung cancer), soft tissue sarcoma, fibrosarcoma, osteosarcoma, pancreatic cancer, among others.

[0305] The disclosed approach is suitable as an adjuvant / neoadjuvant system. For example, during a biopsy, the therapeutic support composition disclosed herein can be placed, and when the test results come back, the doctor can administer the appropriate cocktail to deliver the therapeutic agent to the desired site in the body (a compound of formula (II-A) and optionally another therapeutic agent). The results of the biopsy can indicate the amount and type of therapeutic agent to be delivered to the site of the tumor. For example, chemokines (agents that attract cancer cells and / or immune cells) and adjuvants that boost the immune system with fewer side effects, as well as chemotherapy agents, can be delivered and combined with immunotherapy agents.

[0306] The disclosed compounds and compositions may be administered prior to surgical resection. The disclosed methods may minimize tumor size prior to surgical resection. This may minimize tumor size, particularly for tumors that are surgically resectable. The disclosed compounds and compositions may be administered during surgical resection. The disclosed compounds and compositions may be administered after surgical resection. At the end of surgical resection, the therapeutic support composition may be placed around the resection cavity, and the subject may then be treated with further administration of a therapeutic agent (e.g., pro-doxorubicin) to minimize the risk of any cancer cells that may have been missed at the resection margin.

[0307] The disclosed method may include systemic administration of multiple doses of a functionalized payload that is concentrated at one location. The disclosed method may be used to deliver a second payload. If the tumor is resistant to the first payload, the disclosed method may be used to administer a second functionalized payload. The second payload may be a TCO-labeled payload of gemcitabine or docetaxel. The TCO-labeled payload of gemcitabine or docetaxel may be administered in combination with doxorubicin. The second functionalized payload may be activated by the therapeutic support composition used for the first prodrug.

[0308] The functionalized payloads disclosed herein can act as adjuvants. This combination approach can be beneficial for patients. Chemotherapeutic drugs can treat solid tumors or specific locations, enhance or induce immune responses, and functionalized payloads and / or Augmenting the immunotherapeutic response of other agents may be useful at distant metastatic sites. For example, in certain embodiments, the disclosed compositions and methods can be used with or in conjunction with anthracyclines, taxanes, gemcitabine, and other agents to enhance the efficacy of ipilimumab, nivolumab, pembrolizumab, avelumab (alias: MSB0010718C; Pfizer).

[0309] i. Diffuse pontine glioma The disclosed methods can be used to treat diffuse pontine glioma. Diffuse pontine glioma (DIPG) is a pediatric brainstem tumor that can be extremely malignant and difficult to treat. There is no known therapeutic treatment for DIPG, and the chances of survival remain dismal over the past 40 years. DIPG patients have a median overall survival of only 11 months, with a 2-year survival rate of less than 10%. DIPG accounts for 75-80% of pediatric brainstem tumors, affecting an estimated 200-300 children in the United States each year. Research has been hampered by the rarity of this devastating disease and the lack of a previous experimental model system, and the chances of survival have remained the same over the past 40 years. Diagnosis of DIPG begins with clinical symptoms and can be confirmed by MRI. The disease can begin with months of generalized symptoms, including changes and difficulties in school behavior, double vision, abnormal or restricted eye movements, asymmetric smiling, impaired balance, and weakness. Alternatively, severe neurologic deterioration may occur rapidly, with symptoms present for less than a month before diagnosis. Laboratory examination may reveal a triad of multiple cranial nerve deficits, long tract symptoms such as hyperreflexia and clonus, and ataxia. Elongation of the pontine segments of the brainstem may cause obstructive hydrocephalus and increased intracranial pressure.2

[0310] Nerve nuclei vital to vital functions such as breathing and heartbeat are located in the pons, and if untreated, breathing and heartbeat can be impaired by DIPGs.

[0311] The disclosed methods may be used to deliver molecular payloads to the site of DIPG (e.g., HDAC inhibitors such as panobinostat). The disclosed methods may include systemically administering a drug that is only activated at the tumor site. The disclosed methods may be used as neoadjuvant or adjuvant therapy. During the biopsy, a biomaterial may be placed. The results of the biopsy may indicate the amount and type of therapeutic to be delivered to the site of the tumor. The disclosed compounds and compositions may be administered prior to surgical resection. The disclosed methods may minimize the size of the tumor prior to surgical resection. The disclosed compounds and compositions may be administered during surgical resection. The disclosed compounds and compositions may be administered after surgical resection. At the end of the surgical resection, a biomaterial may be placed around the resection cavity, and the subject may then be treated with additional administrations of a therapeutic agent (e.g., pro-doxorubicin). The disclosed biodegradable gel may be implanted at the time of the biopsy or surgery. The disclosed methods do not require a separate invasive procedure to deliver additional doses of the disclosed compounds and compositions.

[0312] The disclosed method may include systemic administration of multiple doses of a functionalized payload that is concentrated at one location. The disclosed method may be used to deliver a second payload. If the tumor is resistant to the first payload, the disclosed method may be used to administer a second functionalized payload. The second payload may be a TCO-labeled payload of gemcitabine or docetaxel. The TCO-labeled payload of gemcitabine or docetaxel may be administered in combination with doxorubicin. The second functionalized payload may be activated by the therapeutic support composition used for the first prodrug.

[0313] c. Inflammation-related disease or disorder The disclosed methods can be used to treat or prevent diseases and disorders associated with inflammation. Diseases and / or disorders that can be treated and / or prevented by the disclosed methods include, but are not limited to, asthma, arthritis, rheumatoid arthritis, osteoarthritis, autoimmune diseases, autoinflammatory diseases, and the like. These include chronic inflammation, celiac disease, chronic prostatitis, diverticulitis, glomerulonephritis, otitis, necrotizing enterocolitis, inflammatory bowel disease, Crohn's disease, ulcerative colitis, colitis, Behcet's disease, vasculitis, transplant rejection, and autoimmune thyroid disease.

[0314] i. Pigmented villonodular synovitis The disclosed methods may be used to treat pigmented villonodular synovitis. Pigmented villonodular synovitis (PVNS), also known as tenosynovial giant cell tumor (TGCT), is a chronic, progressive tumorigenic process that causes thickening of the synovial lining layer of joints, bursae, or tendon sheaths, overgrows in an aggressive manner, with a very low risk of metastasis. The condition affects approximately 1.8 per million people in the United States, or about 600 people per year, and generally appears in people aged 20-45 years. PVNS can be focal or diffuse. In the diffuse form, the disease process can accelerate wear of tendons and joints, and traditional treatment strategies can have a local recurrence rate of 40-50%. The benign, but aggressive behavior of PVNS makes treatment difficult, as clinicians must weigh the morbidity of treatment against the natural history of the disease process. Methods to locally deliver and activate therapeutic agents may be a solution for conditions such as diffuse PVNS. This limits the systemic side effects of the drug.Diffuse pigmented villonodular synovitis (PVNS), also synonymously called tenosynovial giant cell tumor (TGCT) in the extra-articular manifestations of the disease, is a predominantly localized, aggressive tumorigenic process that affects the synovial lining layer of the joints, bursae, or tendon sheaths, causing it to thicken and overgrow, and inducing a destructive inflammatory process.

[0315] The majority of patients with both localized and diffuse types of PVNS have a gene rearrangement at chromosome 1p11-13, the site of macrophage colony-stimulating factor (CSF-1). The rearrangement results in overexpression of CSF-1, which attracts inflammatory cells that express the CSF-1 receptor (CSF1R) and drives the formation of PVNS.13 CSF-1, a secreted cytokine and hematopoietic growth factor that plays an essential role in the proliferation, differentiation, and survival of monocytes, macrophages, and related cells.

[0316] It has been demonstrated that only a small population of mononuclear stromal cells (2-16%) overexpress CSF-1 in PVNS-affected tissues, and these neoplastic cells constitute a minor component within the tumor. However, most of the cells are non-neoplastic and have high levels of receptor (CSFR1) expression, which is recruited and activated by CSF1 produced by neoplastic cells. Because CSFR1 is a group III receptor tyrosine kinase, tyrosine kinase receptor inhibitors (TKIs) targeting CSF1R (e.g., imatinib, nilotinib, emactuzumab, and PLX3397) are speculated to inhibit PVNS progression, reduce surgical mortality, and preserve patients' quality of life.

[0317] There are at least two types of disease, which can be distinguished histologically. The first, focal PVNS / GCTTS, can appear within joints or around tendon sheaths supporting joints. It can also manifest as a localized extra-articular process, usually affecting the small joints of the hand or wrist (65%-89%) and foot and ankle (5%-15%), or as a localized intra-articular disease, usually affecting the knee, hip, or ankle. The disclosed methods can be used to treat type 1 PVNS / GCTTS. Type 2 PVNS is a diffuse type that affects the entire synovial lining layer. It is most common in the larger joints, usually the hip (4-16%) and knee (66-80%), but can occur in other joints as well (ankle, shoulder, elbow, spine, etc.). This type of disease is more invasive and less successfully treated by surgical excision. The disclosed methods can be used to treat type 2 PVNS.

[0318] Patients with symptomatic, aggressive PVNS, especially the diffuse type, are currently undergoing treatments that have long-term consequences. Concurrent surgical and radiological approaches ultimately result in a benign condition. is too potent. The recent development of systemic drugs with action on the CSF-1R pathway has created an interesting new approach to this frustrating condition. Its use as an adjuvant to surgery has demonstrated promising early results, but side effects remain a limitation. The disclosed method of locally delivering and activating therapeutic agents would readily be beneficial in treating PVNS while avoiding the long-term sequelae of the treatment itself. The disclosed method may eliminate the need for surgery in PVNS patients. The disclosed method may eliminate the need for surgery in focal PVNS. The disclosed method may reduce the recurrence of PVNS. The disclosed method may reduce the local recurrence of diffuse PVNS.

[0319] ii. Arthritis The disclosed treatment method can be used to treat arthritis. Arthritis is a term that can refer to any disorder that affects joints. Symptoms can include joint pain and stiffness. Other symptoms can include redness, warmth, swelling, and reduced range of motion of the affected joint. In some types of arthritis, other organs can also be affected. Onset can be either gradual or sudden. There can be more than 100 types of arthritis. The most common types are osteoarthritis and rheumatoid arthritis. Osteoarthritis occurs with age and can affect the fingers, knees, and hips. Rheumatoid arthritis is an autoimmune disorder that can affect the joints of the hands, joints of the legs, skin, lungs, heart and blood vessels, blood, kidneys, eyes, liver, bones, and nervous system.

[0320] In some embodiments, the disclosed compounds and compositions can be used to treat infections, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmias, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infections, and pigmented villonodular synovitis.

[0321] b. Method of administration The method of treatment may include any number of methods of administering the disclosed compounds or compositions, including tablets, pills, dragees, hard and soft gel capsules, granules, pellets, skin patches, skin creams, skin gels, aqueous, lipid, oily or other solutions, emulsions such as oil-in-water emulsions, liposomes, aqueous or oily suspensions, syrups, elixirs, solid emulsions, solid dispersions or dispersible powders. For the purpose of preparing pharmaceutical compositions for oral administration, the compounds or compositions disclosed herein may be mixed with adjuvants and excipients, such as gum arabic, talcum, starch, sugars (e.g., mannitol, methylcellulose, lactose), gelatin, surfactants, magnesium stearate, aqueous or non-aqueous solvents, paraffin derivatives, crosslinking agents, dispersants, emulsifiers, lubricants, preservatives, flavorings (e.g., ethereal oils), solubility enhancers (e.g., benzyl benzoate or benzyl alcohol) or bioavailability enhancers (e.g., Gelucire®). In the pharmaceutical composition, the compounds or compositions disclosed herein may be dispersed in microparticle, e.g., nanoparticle compositions.

[0322] For parenteral administration, the compounds or compositions disclosed herein may be dissolved or suspended in a physiologically acceptable diluent, such as water, a buffer, an oil with or without a solubilizer, a surfactant, a dispersant, or an emulsifier. Suitable oils may include, for example, olive oil, peanut oil, cottonseed oil, soybean oil, castor oil, and sesame oil. For parenteral administration, the compounds or compositions disclosed herein may be administered in the form of an aqueous, lipid, oily, or other type of solution or suspension, or in the form of liposomes or nano-suspensions.

[0323] The term "parenteral" as used herein refers to modes of administration which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.

[0324] The compounds and compositions disclosed herein may be administered topically. The topical compositions disclosed herein may be applied to the skin of a subject in need thereof. The skin location selected for treatment may be the site of bacterial infection. The skin location selected for treatment may be the skin surrounding the site of infection. The skin location selected for treatment may be the site of bacterial infection and the skin surrounding the site of infection. The skin infection may be due to MRSA. The topical compositions disclosed herein may be applied to the mucosa of a subject in need thereof. The mucosa selected for treatment may be the site of bacterial infection. The mucosa location selected for treatment may be the mucosa surrounding the bacterial infection. The mucosa selected for treatment may be the site of bacterial infection and the mucosa surrounding the site of infection. The mucosa infection may be due to MRSA.

[0325] Topical administration may be performed using a patch containing the compounds and compositions disclosed herein. Topical administration may be performed using a dissolving patch containing the compounds and compositions disclosed herein. Topical administration may be performed using a cream containing the compounds and compositions disclosed herein. Topical administration may be performed using a foam containing the compounds and compositions disclosed herein. Topical administration may be performed using a lotion containing the compounds and compositions disclosed herein. Topical administration may be performed using an ointment containing the compounds and compositions disclosed herein. Topical administration may be performed using a gel containing the compounds and compositions disclosed herein. Topical administration may have fewer side effects than systemic administration of antibiotics.

[0326] In some embodiments, topical compositions comprising a therapeutically effective amount of the compounds and compositions disclosed herein can be applied to infected skin and / or mucosa of a subject to reduce or eliminate infection and / or improve healing of damaged skin and / or mucosa. In certain embodiments, topical compositions comprising a therapeutically effective amount of the compounds and compositions disclosed herein can be applied to sites of skin and / or mucosa infected with MRSA, including infections caused by MRSA biofilms. In these embodiments, the compounds and compositions disclosed herein can be administered alone or in combination with one or more other active agents to reduce infection of the skin and / or mucosa and / or promote healing of the skin and / or mucosa.

[0327] The therapeutic support composition is preferably administered locally to the site of the tumor, such as by injection or implantation. The functionalized payload, such as a compound of formula (IA), (IB), (II-A), or (III-A), may be administered by any convenient route, taking into account the condition of the subject and the judgment of the medical professional. Parenteral administration is the preferred means of administration of the compound of formula (IA), (IB), (II-A), or (III-A).

[0328] The amount of the composition administered to a subject can be determined based on the dosage and / or dosing regimen guidance of the parent drug initially.In general, the composition can enable targeted delivery of the conjugated drug and / or increase its serum half-life, thus enabling at least one of a reduced dose or reduced administration in the dosing regimen.Thus, the composition allows for a reduced dose and / or reduced administration in the dosing regimen compared to the parent drug before conjugation in the composition of the present disclosure.

[0329] The compositions of the present disclosure can be delivered by any suitable means, including oral, parenteral and topical methods. For example, topical transdermal administration methods can be formulated as applicator sticks, solutions, suspensions, emulsions, gels, creams, ointments, pastes, jellies, salves, powders and aerosols.

[0330] The pharmaceutical formulation may be provided in a unit dosage form. In such a dosage form, the pharmaceutical formulation The dosage form may be divided into unit doses containing appropriate amounts of the compositions of the present disclosure. The unit dosage form may be a packaged preparation, the package containing discrete quantities of the preparations, for example, tablets, capsules, and powders, packaged in pouches, vials, or ampoules. The unit dosage form may also be a capsule, tablet, dragee, cachet, or lozenge, or the appropriate number of any of these in packaged form.

[0331] The compositions of the present disclosure may be present in any suitable amount, which may vary depending on a variety of factors including, but not limited to, the weight and age of the subject, disease state, etc. Suitable doses of the compositions of the present disclosure include 0.1 mg to 10,000 mg, or 1 mg to 1000 mg, or 10 mg to 750 mg, or 25 mg to 500 mg, or 50 mg to 250 mg. For example, suitable doses of compositions of the present disclosure include 1 mg, 5 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1000 mg.

[0332] In some embodiments, multiple doses of the composition are administered. The frequency of administration of the composition may vary depending on any of a variety of factors, such as the severity of symptoms, the condition of the subject, etc. For example, in some embodiments, the composition is administered once a month, twice a month, three times a month, every other week (qow), once a week (qw), twice a week (biw), three times a week (tiw), four times a week, five times a week, six times a week, every other day (qod), every day (qd), twice a day (qid), or three times a day (tid).

[0333] The composition of the present disclosure can be administered at any suitable frequency, interval and period.For example, the composition of the present disclosure can be administered once per hour, or 2 or 3 times or more per hour, once per day, or 2 or 3 times or more per day, or once every 2, 3, 4, 5, 6 or 7 days, so as to achieve the desired dosage level of the subject.When the composition of the present disclosure is administered more than once per day, representative intervals include 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes and 60 minutes, and 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours and 24 hours. The compositions of the present disclosure may be administered once, twice, or three or more times for 1 hour, 1-6 hours, 1-12 hours, 1-24 hours, 6-12 hours, 12-24 hours, 1 day, 1-7 days, 1 week, 1-4 weeks, 1 month, 1-12 months, 1 year or more, or indefinitely.

[0334] The composition of the present disclosure can be co-administered with another active agent. Co-administration includes administering the composition of the present disclosure and the active agent within 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 16 hours, 20 hours, or 24 hours of each other. Co-administration also includes administering the composition of the present disclosure and the active agent simultaneously or nearly simultaneously (e.g., within about 1 minute, 5 minutes, 10 minutes, 15 minutes, 20 minutes, or 30 minutes of each other), or sequentially in any order. In addition, the composition of the present disclosure and the active agent can each be administered once a day, or two, three times a day, or more, to achieve the desired daily dosage level.

[0335] The combined administration may involve combined implantation or combined injection.

[0336] In some embodiments, the combined administration may involve co-formulation, e.g., preparation of a single pharmaceutical formulation that includes both the composition of the present disclosure and the active agent, hi other embodiments, the composition of the present disclosure and the active agent may be formulated separately and administered combined to a subject.

[0337] The composition of the present disclosure and the active agent may be present in the formulation in any suitable weight ratio, for example, 1:100-100:1 (w / w), or 1:50-50:1, or 1:25-25:1, or 1:10-10:1, or 1:5-5:1 (w / w). The composition of the present disclosure and the other active agent may be present in any suitable weight ratio, for example, 1:100 (w / w), 1:75, 1:50, 1:25, 1:10, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 10:1, 25:1, 50:1, 75:1, or 100:1 (w / w). Other doses and dose ratios of the composition of the present disclosure and the active agent are also suitable in the formulations and methods described herein.

[0338] C. Combination therapy In one aspect, the present invention provides a method of treating cancer or enhancing or eliciting an immune response, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt or composition thereof; a therapeutic support composition as described herein; and a therapeutically effective amount of one or more immunomodulatory agents, or a pharma- ceutically acceptable salt thereof.

[0339] The present invention also provides pharmaceutical combinations comprising a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; a therapeutic support composition as described herein; and one or more immunomodulatory agents for use in the treatment or prevention of a disease or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or inducing an immune response. The present invention also provides pharmaceutical combinations comprising a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; a therapeutic support composition as described herein; and one or more immunomodulatory agents for use in a method for treating or preventing a disease or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in a method for enhancing or inducing an immune response.

[0340] The invention also provides for the use of a pharmaceutical combination comprising: a) a compound of Formula (II-A) or (III-A), or a pharma- ceutically acceptable salt, or composition thereof; a therapeutic support composition; and one or more immunomodulatory agents in the manufacture of a medicament for the treatment or prevention of a condition or disorder, such as cancer, an infectious disease, tissue injury, stenosis, ischemia, revascularization, myocardial infarction, arrhythmia, vascular occlusion, inflammation, autoimmune disorders, transplant rejection, macular degeneration, rheumatoid arthritis, osteoarthritis, periprosthetic infection, and pigmented villonodular synovitis; or for use in enhancing or eliciting an immune response.

[0341] In the methods and uses described herein, pharmaceutical combinations comprising a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt thereof, or composition; a therapeutic support composition; and one or more immunomodulators may be administered / used simultaneously, separately, or sequentially, and in any order, and the components may be administered separately or as a fixed combination. For example, delaying disease progression or treating a disease according to the present invention may involve administration of a first active ingredient, in free or pharma- ceutically acceptable salt form, and a second active ingredient, in free or pharma- ceutically acceptable salt form, simultaneously or sequentially in any order, in a jointly therapeutically effective or effective amount, e.g., in a daily dose corresponding to the amounts described herein. The individual active ingredients of the combination may be administered separately at various times during the treatment period, or simultaneously in a divided or single dosage form. The present invention should therefore be understood as embracing all such regimes of simultaneous or alternating treatment, and the term "administering" should be interpreted accordingly. Thus, a pharmaceutical combination, as used herein, refers to a combination administration. The term "combination" refers to a combination of a compound of formula (II-A) or (III-A) that is administered orally or in a single unit dosage form for administration, where the combination may be administered independently at the same time or at different times. As another example, the compound of formula (II-A) or (III-A) may be administered / used simultaneously (e.g., by co-injection or co-implantation), separately, or sequentially after the therapeutic support composition and one or more immunomodulatory agents.

[0342] The method and use in the treatment of cancer includes administering / localizing a therapeutic support composition to a tumor. In the method and use disclosed herein, the administration of a compound of formula (II-A) or (III-A), or a pharma- ceutically acceptable salt or composition thereof; a therapeutic support composition; and one or more immunomodulatory agents can inhibit tumor growth.

[0343] Any of the methods and uses, including combination therapies, disclosed herein using formula (I), (IA), (II-A) or (III-A) may be further combined with another therapeutic agent, such as an anti-cancer agent, an antibacterial agent, an immunomodulatory agent, and a vaccine.

[0344] The other therapeutic agent may be an anti-cancer drug, which may be any of the anti-cancer drugs described herein as anti-cancer payload drugs of formula (IB) or (II-A).

[0345] Another therapeutic agent may be a vaccine, including an adjuvant and / or an antigen.

[0346] Another therapeutic agent may be a TLR or STING agonist as described herein for Formula (I) / (IA).Other immunomodulatory agents include cytokines, chemokines, chemokine antagonists, and immune checkpoint inhibitors.

[0347] Cytokines can limit tumor cell proliferation directly through antiproliferative or proapoptotic activity, or indirectly by stimulating the cytotoxic activity of immune cells against tumor cells. Cytokines that can be used as immunomodulatory agents include, but are not limited to, IFN-α, IFN-β, and IFN-γ, interleukins (e.g., IL-1 through IL-29, particularly IL-7, IL-12, IL-15, IL-18, and IL-21), tumor necrosis factors (e.g., TNF-α and TNF-β), erythropoietin (EPO), MIP3a, ICAM, macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (GCSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF), as described in US Patent Application Publication No. 2008 / 0014222. In embodiments of the invention, the cytokine is IL-2, IL-2 covalently bound to an immunoglobulin (e.g., sergituzumab amnaleukin, RO6874281) or a PEG molecule (e.g., NKTR-214), IL-10, PEGylated IL-10 (e.g., pegylodecakin), IL-12, IL-15, recombinant aglycosylated IL-15, a fusion protein of IL-15 with the binding domain of IL-15Rα (e.g., RLI), a triple fusion protein of human IL-15, the binding domain of IL-15Rα and apolipoprotein AI, ALT-803 (il-15 fused to an IgG1 Fc domain), IL-21, GM-CSF, talimogene laherparepvec, IFN-α, pegylated IFN-α, apolipoprotein AI fusion protein with IFN-α.

[0348] Inhibitors of certain cytokines, their cognate receptor agonists and / or antagonists may also be used in cancer therapy. Cytokines are secreted or membrane-bound proteins that act as mediators of intracellular signaling to regulate the homeostasis of the immune system. They are produced by cells of the innate and adaptive immunity in response to microorganisms, self-antigens and tumor antigens. Inhibitors of TNF-α (e.g., infliximab, certolizumab) inhibit the inhibition of TGF-β (e.g., galunisertib, fresoxofenib), especially in relation to PD-1 pathway blockade. Limumab, M7824), and CSF-1 (e.g., pexidartinib, cabilalizumab) can be used in the methods of the invention.

[0349] Immunotherapy using cytokines and cytokine inhibitors has been described by Berraondo et al. al., British Journal of Cancer (2019) 120, 6-15, incorporated herein by reference.

[0350] Cytokine and / or chemokine receptor inhibitors may be used as immunomodulatory agents; these are chemotactic proteins that have the potential to attract macrophages, T cells, eosinophils, basophils, and other cells to sites of inflammation, infection, and / or tumor growth. These proteins are usually of low molecular weight (7-9 kD). Chemokines form four structural subclasses (C, CC, CXC, and CX3C) classified according to their primary amino acid structure, which contain various combinations of conserved cysteine ​​residues.

[0351] Potentially suitable immune modulating chemokines are CCL27 and CCL28, CC (CCL2, CCL3, CCL5) and CXC (CXCL1, CXCL2, CXCL5, CXCL6, CXCL8, CXCL9, CXCL10, CXL12).

[0352] ELRCXC chemokines such as IL-8, GROα, GROβ, GROγ, NAP-2, and ENA-78 (Strieter, 1995, J Biol Chem, 270:27348-57) have also been implicated in the induction of tumor angiogenesis (new blood vessel growth). The angiogenic activity is due to the binding of ELRCXC-chemokines to CXCR2, and in the case of IL-8, possibly CXCR1, which are expressed on the surface of vascular endothelial cells (ECs) in peripheral blood vessels, and their activation. Many different types of tumors have been shown to produce ELRCXC chemokines. Chemokine production correlates with a more aggressive phenotype (Inoue, 2000, Clin Cancer Res, 6:2104-2119) and poor prognosis (Yoneda, 1998, J Nat Cancer Inst, 90:447-54). Chemokines are potent chemotactic factors, and in particular the ELRCXC chemokines have been shown to induce EC chemotaxis. Thus, these chemokines are thought to induce chemotaxis of epithelial cells toward their site of production in tumors. This may be a key step in the induction of angiogenesis by tumors. Inhibitors of CXCR2 or dual inhibitors of CXCR2 and CXCR1 would inhibit the angiogenic activity of ELRCXC chemokines and thus block tumor growth. This antitumor activity has been demonstrated for antibodies against IL-8 (Arenberg, 1996, J Clin Invest, 97:2792-802), ENA-78 (Arenberg, 1998, J Clin Invest, 102:465-72), and GROα (Haghnegandar, 2000, J Leukoc Biology, 67:53-62). CXC chemokine inhibitors, as described in U.S. Pat. No. 10,046,002 (hereby incorporated by reference), include the following: [ka] Includes.

[0353] Suitable immunomodulatory agents for use in the methods of the invention include chemokine or chemokine receptor antagonists that inhibit the recruitment of suppressive immune cells into the tumor microenvironment. For example, but not limited to, the methods of the invention may use CCR1, CCR2 or CCR5 antagonists that reduce the infiltration of myeloid suppressor cells and regulatory T cells.

[0354] Suitable CCR, CXCR, and CCL inhibitors include the following: CCR1 (e.g., CCX721, BL5923), CCR2 (e.g., CCX9588, PF-04136309, CCX872, RDC018, 747, iCCR2), CCL2 (e.g., CNTO 888), CCR4 (e.g., Affi 5, AF399 / 420 / 1802), CCR5 (e.g., Maraviroc), CCR7 (e.g., siRNA, MSM R707 ), CXCR2 (e.g., Navarixin, SB225002, Reparixin, SB265610, AZD5069), CXCR4 (e.g., AMD3100, AMD3465, LY2510924, BKT140, BMS-936564, PF-06747143, PRX177561, POL5551, USL311, CTCE-9908).

[0355] Immune checkpoint inhibitors include, but are not limited to, PD-1 inhibitors (e.g., nivolumab, pembrolizumab, pidilizumab, sintilimab, AMP-224), PD-L1 inhibitors (e.g., atezolizumab, avelumab, durvalumab, BMS-936559), CTLA4 inhibitors (e.g., ipilimumab, tremelimumab), IDO inhibitors (e.g., indoximod, epacadostat), TIGIT inhibitors (e.g., LAG-3, e.g., the anti-LAG-3 antibodies described in U.S. Patent Application Publication No. 2015 / 0259420, which is incorporated herein by reference; TIM-3, e.g., the anti-TIM-3 antibodies described in U.S. Patent Application Publication No. 2015 / 0218274, which is incorporated herein by reference), and BTLA pathway antagonists.

[0356] In some embodiments, the immune response is modulated using xenobiotics, biologics, natural or artificial adjuvants, cell therapy, and / or checkpoint inhibitors, including inhibitors of PD-1, PD-L1, CTLA-4, B7 molecules, TIGIT, Tim-3 and / or Lag-3, indoleamine 2,3-dioxygenase (IDO).

[0357] Another therapeutic agent may be an immune checkpoint inhibitor, such as a PD-1 inhibitor (e.g., nivolumab, pidilizumab, sintilimab), a PD-L1 inhibitor (e.g., atezolizumab, avelumab, durvalumab, BMS-936559), a CTLA4 inhibitor (e.g., ipilimumab, tremelimumab), or an IDO inhibitor (e.g., indoximod, epacadostat).

[0358] The other therapeutic agent may be a compound of formula (IB), or a pharma- ceutically acceptable salt thereof.

[0359] For the treatment of bacterial infections, the compounds and compositions may be combined with various antibiotics, including, but not limited to, vancomycin, linzolid, teicoplanin, ceftaroline, clindamycin, mupirocin, trimethoprim-sulfamethoxazole, tetracycline, daptomycin, sulfa drugs, ceftobiprole, ceftaroline, dalbavancin, telavancin, torezolid, iclaprim, nemonoxacin, platensimycin, and oxadiazoles.

[0360] The compounds and compositions may be combined with agents that inhibit bacterial biofilm formation, including, but not limited to, imidazole derivatives, indole derivatives, emodin, flavonoids, ginger extract, Hypericum perforatum, 7-epiclusianon, isolimonic acid, tannic acid, chelerythrine, carvacrol, bugaine, resveratrol, garlic extract, natural halogenated furanones, brominated alkylidene lactams, and AHL-based inhibitors.

[0361] The compounds and compositions may be combined with lysine-conjugated aliphatic norspermidine analogs. The compounds and compositions may be combined with phage therapeutics. In the case of infections associated with a medical device or prosthesis, the compounds and compositions may be administered in conjunction with removal of the medical device or prosthesis. A new sterile medical device or prosthesis may be implanted in the subject.

[0362] The compounds and compositions may be combined with agents that mitigate potential side effects from antibacterial agents, including, but not limited to, probiotics, antidiarrheal agents, antiemetics, and painkillers that may mediate or treat side effects.

[0363] Subjects may be undergoing various treatments for comorbid conditions.

[0364] The other therapeutic agent may be administered simultaneously or sequentially with the disclosed compounds and compositions. Sequential administration includes administration before or after the disclosed compounds and compositions. The other therapeutic agent may be administered before the disclosed compounds and compositions. The other therapeutic agent may be administered after the disclosed compounds and compositions. The other therapeutic agent may be administered simultaneously with the disclosed compounds and compositions. In some embodiments, the other therapeutic agent may be administered in the same composition as the disclosed compounds. In other embodiments, there may be a time interval between the administration of the other therapeutic agent and the administration of the disclosed compounds or compositions. In some embodiments, administration of the disclosed compounds or compositions with the other therapeutic agent may allow for lower doses of the other therapeutic agent and / or administration at less frequent intervals. When used in combination with one or more other active ingredients, the compounds or compositions of the present invention and the other active ingredients may be used in lower doses than when each is used alone. Thus, pharmaceutical compositions of the present invention include those that contain one or more active ingredients in addition to the compounds of the present disclosure.

[0365] 5. Kit Aspects of the present disclosure include kits having the compositions described herein.

[0366] The kit may include a compound of formula (IA), or a pharma- ceutically acceptable salt thereof, or composition, and a therapeutic support composition. The kit may include a compound of formula (IA), or a pharma- ceutically acceptable salt thereof, or composition, and a compound of formula (IB), formula (II-A), or formula (III-A), or a pharma- ceutically acceptable salt thereof, or composition.

[0367] The kit may include a compound of Formula (II-A) or Formula (III-A), or a pharma- ceutically acceptable salt thereof, or composition, and one or more immunomodulatory agents, or a pharma- ceutically acceptable salt thereof, or composition, and, optionally, a therapeutic support composition. The kit may include a therapeutic support composition described herein, and one or more immunomodulatory agents, or a pharma- ceutically acceptable salt thereof, or composition.

[0368] The therapeutic support composition, one or more immunomodulatory agents, and compounds of Formula (IA), (IB), (II-A), and / or Formula (III-A) may be in separate containers within the packaging. One or more therapeutic support compositions may be provided in a kit.

[0369] The kits described herein may include packaging configured to contain a composition (e.g., a therapeutic support composition and / or one or more immunomodulatory agents). Similarly, one or more compounds of Formula (IA), (IB), (II-A), and / or Formula (III-A) may be provided in the kit. The packaging may be a sealed packaging, such as a sterile sealed packaging. "Sterile" means substantially free of microorganisms (e.g., fungi, bacteria, viruses, spore forms, etc.). In some cases, the packaging may be configured to be sealed, e.g., a moisture-resistant packaging, optionally under air-tight and / or vacuum seal.

[0370] In certain embodiments, the kit includes a reagent that can be used as a releasing agent for the releasable linkers described herein. The releasing agent can be any of the releasing agents described herein, including, but not limited to, chemical releasing agents (e.g., acids, bases, oxidizing agents, reducing agents, etc.). ), solvents, etc. The release reagent in the kit can be provided in any convenient form, such as a gas, a solution, a solid, granules, a powder, a suspension, etc. The release reagent may be packaged in a container separate from the compositions in the kit.

[0371] In addition to the above components, the subject kits may further include instructions for carrying out the subject methods. These instructions may be present in the subject kits in a variety of forms, one or more of which may be present in the kit. One form in which these instructions may be present is as information printed on a suitable medium or support, such as one or more sheets of paper on which the information is printed, in the kit's packaging, a package insert, or the like. Another form of the instructions may be a computer readable medium on which the information is recorded or stored, such as a CD, DVD, Blu-ray, computer readable memory (e.g., flash memory), and the like. Yet another form of the instructions may be a website address, which allows the information to be viewed at the removed site via the Internet. Any convenient means may be present in the kit.

[0372] 6. Working Example The present disclosure has many aspects, exemplified by the following non-limiting examples. The following examples are provided to provide one of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the following experiments are all experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. "Average" means arithmetic mean. Standard abbreviations may be used, such as bp (base pairs); kb (kilobase); pl (picoliters); s or sec (seconds); min (minutes); h or hr (hours); aa (amino acids); kb (kilobase); bp (base pairs); nt (nucleotides); im (intramuscular); ip (intraperitoneal); sc (subcutaneous), etc.

[0373] Many general references are available that provide well-known chemical synthetic schemes and conditions useful for synthesizing the disclosed compounds (see, for example, Smith and March, March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, 2001; or Vogel, A Textbook of Practical Organic Chemistry, Including Qualitative Organic Analysis, Fourth Edition, New York: Longman, 1978).

[0374] The compounds described herein can be purified by any purification protocol known in the art, such as HPLC, preparative thin layer chromatography, flash column chromatography, and ion exchange chromatography. Any suitable stationary phase can be used, including normal and reverse phase and ionic resins. In certain embodiments, the disclosed compounds are purified via silica gel and / or alumina chromatography. See, for example, Introduction to Modern Liquid Chromatography, 2nd Edition, ed. LRSnyder and JJKirkland, John Wiley & Sons, 1999. and Sons, 1979; and Thin Layer Chromatography, ed E. Stahl, Springer-Verlag, New York, 1969.

[0375] During any step of the preparation of the subject compounds, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules concerned. This can be achieved using conventional protecting groups as described in standard works such as: JFW MacOmie, "Protective Groups in Organic Chemistry", Plenum Press London and New York 1973; TW Greene and PG M Huts, "Protective Groups in Organic Synthesis", Third edition, Wiley, New York 1999, in "The Peptides"; Volume 3 (editors: E. Gross and J. Meienhofer), Academic Press, London and New York 1981, in "Methoden der organischen Chemie", Houben-Weyl, 4 th edition, Vol. 15 / l, Georg Thieme Verlag, Stuttgart 1974, in H.-D. Jakubke and H. Jescheit, “Aminosauren, Peptide, Protein”, Verlag Chemie, Weinheim, Deerfield Beach, and Basel 1982, and / or Jochen Lehmann, "Chemie der Kohlenhydrate: Monosaccharide and Derivate", Georg Thieme Verlag, Stuttgart 1974. The protecting groups may be removed at a later convenient stage using methods known in the art.

[0376] The subject compounds can be synthesized through a variety of different synthetic routes using commercially available starting materials and / or starting materials prepared by conventional synthetic methods.The following schemes illustrate various examples of synthetic routes that can be used to synthesize the compounds disclosed herein.The compound of formula (IA) can be prepared according to the synthetic procedure for preparing the compound of formula (IB).

[0377] The following abbreviations are used herein: ACN Acetonitrile Boc tert-butoxycarbonyl Cy5 Cyanine 5 Cy5.5 Cyanine 5.5 Dapto (dapt) Daptomycin DCC N,N'-dicyclohexylcarbodiimide DCM Dichloromethane dd Double distillation DIBAL Diisobutylaluminum hydride DIPEA Diisopropylethylamine DMAP 4-Dimethylaminopyridine DMFN, N-Dimethylformamide DMSO Dimethyl sulfoxide Doxorubicin EDCI N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride Et Ethyl EtOAc Ethyl acetate FCC Flash Column Chromatography Fmoc Fluorenylmethyloxycarbonyl h or hr Time HA Hyaluronic Acid HAT Tetrazine-modified hyaluronic acid HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate salt HBTU Hexafluorophosphate Benzotriazole Tetramethyluronium HMT Hydrogel modified tetrazine HOAt 1-Hydroxy-7-azabenzotriazole HOBt 1-Hydroxybenzotriazole HPLC High Performance Liquid Chromatograph iPrOH Isopropyl Alcohol LCMS Liquid Chromatography-Mass Spectrometry Me Methyl MeCN Acetonitrile MeOH Methanol MES 2-(N-morpholino)ethanesulfonic acid MeTz Methyltetrazine min MTD maximum tolerated dose NHS N-hydroxysuccinimide NMPN-Methylpiperazine PBS Phosphate Buffered Saline Ph Phenyl ppm parts per million pyridine rt / RT room temperature SEM Standard error of the sample mean Sulfo-NHS N-hydroxysulfosuccinimide TAG tetrazine modified activated gel TBME tert-Butyl methyl ether TCO trans-cyclooctene TEA Triethylamine THF Tetrahydrofuran TLC Thin Layer Chromatography TFA Trifluoroacetic acid TsCl Tosyl chloride or toluenesulfonyl chloride UVLVG Ultra-high purity low viscosity guluronic acid Vanco Vancomycin EXAMPLES

[0378] Example A1 Acid-TCO-doxorubicin (axial isomer) rel-(1R,4E,6R,pS)-6-Hydroxy-1-methylcyclooct-4-ene-1-carboxylic acid (axial isomer 2) A solution of 5.34 g (95.2 mmol) of potassium hydroxide in 16.7 mL of water was added over 5 min to a water-cooled solution of the trans-cyclooctene ester 1 isomeric mixture (Rossin et.al., Bioconjugate Chem., 2016, 27, 1697-1706) (1.64 g, 8.28 mmol, axial / equatorial isomer ratio approximately 1.2:1 for this specific batch) in 37 mL of methanol. The solution was stirred at room temperature for 18 h. Water (51 mL) was added and the mixture was extracted with 3 × 150 mL of TBME. The combined organic layers were washed with 100 mL of water and then dried under vacuum to obtain the unhydrolyzed equatorial ester 1b. The combined aqueous layers were washed with 300 mL of TBME. The mixture was treated with TBME and then 15 g of citric acid. The layers were separated and the aqueous layer was extracted with TBME (3×150 mL). The combined organic layers were dried and then rotary evaporated at 25° C. to give 873 mg (57%) of the pure axial isomer 2 of trans-cyclooctenoic acid as a colorless oil. 1 H-NMR (CDCl3): δ=6.15-5.95(m,1H),5.6(d,1H),4.45(bs,1H),2.4-1.7(m,7H),1.6(dd,1H),1.18(s,3H). 13 C-NMR (CDCl3): δ=185.4(C=O),134.8(=CH),130.7(=CH),69.8(CH),44.8,38.2,31.0,29.8(CH2),18.1(CH3).

[0379] rel-(1R,4E,6R,pS)-2,5-dioxopyrrolidin-1-yl-6-((((2,5-dioxopyrrolidin-1-yl)oxy)carbonyl)oxy)-1-methylcyclooct-4-ene-1-carboxylate (axial isomer 3) To a solution of compound 2 (873 mg, 4.74 mmol) in 24.0 mL of MeCN, DIPEA (4.59 g, 35.6 mmol) was added followed by N,N'-disuccinimidyl carbonate (5.22 g, 20.4 mmol). The mixture was stirred at RT for 3 days and then evaporated under vacuum at 25°C. The residue was chromatographed on 40 g silica using dichloromethane as eluent and then eluted with dichloromethane containing increasing amounts of TBME (0-20%). The product fractions were combined and dried under vacuum. The resulting residue was stirred with TBME until a homogenous suspension was obtained. Filtration and washing gave 761 mg of product 3 as a white solid (38%); 1 H-NMR(CDCl3):δ=6.07(ddd,J=16.8,10.7,3.5Hz,1H),5.62(dd,J=16.7,2.5Hz, 1H),5.25(s,1H),2.83(2s,8H),2.5-2.25(m,4H),2.2-1.9(m,4H),1.28(s,3H).

[0380] NHS-TCO-doxorubicin (axial isomer 4) Doxorubicin hydrochloride (53.7 mg; 0.093 mmol) and 3 (39.1 mg; 0.093 mmol) were dissolved in DMF (2.0 mL) and then DIPEA (60.1 mg; 0.465 mmol) was added. The solution was stirred at room temperature under an argon atmosphere for 22 h. HPLC analysis showed about 60% of the desired product with a double peak. The crude product was divided into two portions. One portion was treated with morpholine (4.0 mg, 0.047 mmol, 5 eq) at room temperature for 24 h. The reaction was stirred at room temperature for an additional 20 h as starting material was still present. The conversion was about 71%. The product also showed a double peak. The product was purified by preparative HPLC to give fairly pure product. The product was confirmed by LCMS to have m / z 935.9 (M+114-1). Another portion was treated with 1-methylpiperazine (4.7 mg, 0.047 mmol, 5 eq) at room temperature for 24 h. Starting material was still present so the reaction was stirred at room temperature for an additional 20 h. Conversion was about 64%. The product also showed a double peak. The product was purified by preparative HPLC to give fairly pure product. The product was confirmed by LCMS to have m / z 948.9 (M+114-1).

[0381] NHS-TCO-doxorubicin (axial isomer 4) Doxorubicin hydrochloride (1.05 g; 1.8 mmol) and 3 (761 mg; 1.8 mmol) were dissolved in DMF (18 mL), followed by the addition of DIPEA (1.16 g; 9.0 mmol). The solution was stirred at room temperature under nitrogen atmosphere for 22 h. HPLC analysis showed that the reaction had proceeded well, with the product having a single peak. The remaining crude product was concentrated to dryness on a rotavapor to remove DMF. The residue was purified by FCC (iPrOH / DCM: 0%-23%) to give the pure product 4 (1.015 g, 66%) as a red solid. 1 H-NMR(CDCl3):δ=13.97(s,1H),13.22(s,1H),8.03(d,J=7.9Hz,1H),7.78(t,J=8.0Hz, 1H),7.38(d,J=8.6Hz,1H),5.85(m,1H),5.59(m,1H),5.51(s,1H),5.29(s,1H).5.16(d, J=8.4Hz,1H),5.12(s,1H),4.75(d,J=4.8Hz,2H),4.52(d,J=5.8Hz,1H),4.15(q,J=6.5Hz ,1H),4.08(d,J=3.6Hz,3H),3.87(m,1H),3.69(m,1H),3.26(d,J=18.8Hz,1H),3.00(m,2H ),2.81(s,4H),2.4-1.7(br.m,13H),1.62(s,2H),1.30(d,J=6.5Hz,3H),1.23(s,3H)ppm. [ka]

[0382] Intermediate 4 (approximately 2.4 mg) in DMF (0.10 mL) can be treated with saturated sodium bicarbonate (0.10 mL) at room temperature. After 18 h, most of the starting material was consumed and the reaction was still complex. The crude product could be purified by Prep HPLC to give fairly pure product.

[0383] Example A2 [ka] General procedure for the preparation of TCO-pexidartinib To a solution of pexidartinib (PLX3397) (373 mg, 0.89 mmol) in DMF (4.0 mL) at 0 °C, sodium hydride (~60%, 39 mg, ~0.96 mmol) was added; the reaction mixture was stirred under nitrogen for 1 h, followed by the addition of TCO-PNB ester (200 mg, 0.68 mmol). The resulting mixture was stirred at rt overnight and evaporated under vacuum. The reaction mixture was diluted with water (30 mL) and then extracted with ethyl acetate (2 x 30 mL). The combined organic layers were washed with brine, dried (Na2SO4), and evaporated under vacuum. The residue was purified by flash chromatography on silica gel eluting with dichloromethane followed by MeOH-CH2Cl2 (0-5%) to give TCO-pexidartinib (145 mg, 37%). LC-MS: 571 [M+H] +1 H NMR(300MHz,CDCl3)δ 8.72(s,1H),8.41(s,1H),8.05(s,1H),7.85(d,J=6.9Hz,1H),7.66(s,1H),7.62(d,J= 7.8Hz,1H),7.56(s,1H),7.29(d,J=2.4Hz,1H),6.37(d,J=8.4Hz,1H),6.15(m,1H),5. 74(s,1H),5.60(d,J=6.0Hz,1H),4.88(t,J=6.0Hz,1H),4.67(d,J=6.0Hz,2H),3.87(s,1H),2.50(m 1H),2.30(m,1H),2.10 -0.80(m,8H).

[0384] Example A3 [ka] General Procedure for Preparation of TCO-Valdecoxib To a solution of valdecoxib (157 mg, 0.5 mmol) in DMF (4 mL) was added TCO-PNB ester (129 mg, 0.44 mmol), DMAP (106 mg, 0.88 mmol). The mixture was stirred at rt for 40 h, diluted with ethyl acetate (100 mL), washed with brine (40 mL), dried over sodium sulfate, and evaporated under vacuum. The product was purified by flash chromatography on silica gel eluting with DCM, then MeOH-DCM (5%) to obtain compound TCO-valdecoxib (201 mg, 97%) as a white solid. LC-MS: 467 [M+H] + . 1 H NMR(300MHz,CDCl3)δ 8.03(d,J=8.7Hz,2H),7.65(m,1H),7.43-7.32(m,7H),5.73(m,1H),5.64(d ,J=16.5Hz,1H),5.33(s,1H),2.50(s,3H),2.43(m,1H),2.09-0.77(m,9H).

[0385] Example A4 [ka] General Procedure for Preparation of TCO-Celecoxib A solution of celecoxib (141 mg, 0.37 mmol) in DMF (4 mL) was added to TC O-PNB ester (100 mg, 0.34 mmol), DMAP (106 mg, 0.88 mmol) were added. The mixture was stirred for 40 h, diluted with ethyl acetate (100 mL), washed with water (30 mL) and brine (30 mL), dried over sodium sulfate, and concentrated in vacuo. The product was purified by flash chromatography on silica gel eluting with methanol (5%) in DCM to give TCO-celecoxib (162 mg, 88%). LC-MS: 534 [M+H] + 1 H NMR(300MHz,CDCl3)δ 8.01(d,J=8.7Hz,2H),7.60(br,1H),7.50(d,J=8.7Hz,2H),7.18(d,J=8.1Hz,2H),7.14(d,J=8 .1Hz,2H),6.74(s,1H),5.69(m,1H),5.45(d,J=12.0Hz,1H),5.30(s,1H),2.44(m,1H),2.38(s 3H),2.03-0.76(m,9H).

[0386] Example A5 Synthesis of TCO-monomethylauristatin E (TCO-MMAE) conjugates [ka] Preparation of TCO-MMAE conjugates To monomethyl auristatin E (170 mg, 0.24 mmol) in DMF (2 mL) at rt was added TCO-bis-NHS (100 mg, 0.24 mmol) and DIPEA (93 mg, 0.72 mmol). The solution was stirred at rt for 20 h, acetonitrile (ACN, 8 mL) was added, and the mixture was purified by prep-HPLC (ACN / water 0-100%, formic acid 0.1%) to obtain TCO-NHS-MMAE (88 mg, 36%). To TCO-NHS-MMAE (78 mg, 0.076 mmol) in THF (2 mL) and HO (2 mL) at rt was added LiOH (9.2 mg, 0.38 mmol). The solution was stirred at rt for 20 h. After removal of the solvent, HCl (aq, 0.5 N) was added until pH approx. 3. The mixture was analyzed by prep-HPLC (ACN / water 0–100%, formic acid 0.1%) ) to give TCO-acid-MMAE (54 mg, 76%, two isomers). LCMS: (ESI+) 928 [M+H].

[0387] Example A6 Synthesis of trans-cyclooctene (TCO)-glycine-doxorubicin conjugate [ka] Preparation of TCO-glycine-doxorubicin conjugate To a solution of doxorubicin hydrochloride (100 mg) in 1 mL DMSO was added TCO-Bis-NHS (75 mg). DIPEA (148 μL) was added by injection. After the mixture was stirred overnight, glycine (51 mg) was added to the reaction in one portion and the reaction was stirred for 24 h. The mixture was diluted with 2 mL HO and then purified by HPLC to obtain TCO-Gly-Dox. MS: (ESI+) 833 [M+Na].

[0388] Example A7 Antibiotic-TCO conjugates Example A7A Dapto-TCO-Amino Acid Synthesis [ka] Example protocol: Add daptomycin (100 mg, 0.062 mmol), TCO-bis-NHS (62.5 mg, 0.149 mmol), and triethylamine (62.5 μL, 45.3 mg, 0.448 mmol) to DMSO and stir at RT overnight to produce Dapto-TCO-NHS. LCMS: (ESI-) 1926.8 [MH]. Add aspartic acid (104.5 mg, 0.785 mmol) and 4-dimethylaminopyridine (150.9 mg, 1.235 mmol) to Dapto-TCO-NHS (126.1 mg, 0.0654 mmol) and stir at 37 °C for 18 h. Purify by HPLC to obtain Dapto-TCO-Aspartic acid. Yield: 100 mg, 0.0514 mmol. LCMS:(ESI-)1944.8[MH].

[0389] Using this approach, glycine and aspartic acid modified TCO-prodrugs were generated, which can be generally applied to the incorporation of other amino acid cargoes.

[0390] Example A7B Daptomycin-TCO-glycine conjugate [ka] Daptomycin (537 mg, 0.33 mmol), TCO-bis-NHS (350 mg, 0.83 mmol), and triethylamine (0.350 mL, 2.51 mmol) in DMSO (11 mL). Stir overnight at RT. Then heat to 37 °C. Add glycine (300 mg, 4.00 mmol) and triethylamine (1.8 mL, 13 mmol) and stir for 18 h. Add 8 mL water and purify by HPLC. Yield: Dapto-TCO-Glycine-373 mg, 0.20 mmol, 59.6%.

[0391] Example A7C Vanco-bis-TCO-glycine conjugate [ka] Example A7C can be synthesized using a protocol similar to that of Example A7B. Vanco-bis-TCO-glycine, tested up to 64 μg / ml (32 μM), exhibits no activity against bacteria as measured by microcalorimetry, indicating drug inactivation after modification.

[0392] Typical HPLC purification conditions for TCO amino acid conjugates are as follows: Column: Higgins Cat#PS-253C-C185, 250 x 30 mm, Phalanx C18 5 μm Solvent A: Water (0.1% formic acid) Solvent B: Acetonitrile (0.1% formic acid)

[0393] [Table 1]

[0394] Example A8 Hyaluronic acid modified tetrazine [ka] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH=6.5), 0.0500 grams of sodium hyaluronate (200 kDa) was added and stirred until it dissolved (4 hours). To this, N-hydroxysulfosuccinimide (23.3 mg, 0.107 mmol), N,N'-dicyclohexylcarbodiimide (42.0 mg, 0.219 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (15.9 mg, 0.066 mmol) were added. The reaction mixture was stirred in the dark for 20 hours, after which it was quenched with hydroxylamine (66.2 mg, 0.953 mmol). The hyaluronic acid product was purified in the dark against deionized water containing decreasing concentrations of salt (NaCl, 0.13 M to 0.0 M) over a period of 5 days. The hyaluronic acid product was filtered (0.22 μm) and lyophilized for 5 days.

[0395] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH=6.5), 0.0500 grams of sodium hyaluronate (100 kDa) was added and stirred until it dissolved (4 hours). To this, N-hydroxysulfosuccinimide (40.6 mg, 0.19 mmol), N,N'-dicyclohexylcarbodiimide (72.1 mg, 0.38 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (28.4 mg, 0.12 mmol) were added. The reaction mixture was stirred in the dark for 20 hours, after which it was quenched with hydroxylamine (117.1 mg, 1.69 mmol). The hyaluronic acid product was purified in the dark against deionized water containing decreasing concentrations of salt (NaCl, 0.13 M to 0.0 M) over a period of 5 days. The hyaluronic acid product was filtered (0.22 μm) and lyophilized for 5 days.

[0396] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH=6.5), 0.0500 grams of sodium hyaluronate (60 kDa) was added and stirred until it dissolved (4 hours). To this, N-hydroxysulfosuccinimide (58.2 mg, 0.27 mmol), N,N'-dicyclohexylcarbodiimide (103.9 mg, 0.54 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (40.4 mg, 0.17 mmol) were added. The reaction mixture was stirred in the dark for 20 hours, after which it was quenched with hydroxylamine (165.7 mg, 2.38 mmol). The hyaluronic acid product was purified in the dark against deionized water containing decreasing concentrations of salt (NaCl, 0.13 M to 0.0 M) over a period of 5 days. The hyaluronic acid product was filtered (0.22 μm) and lyophilized for 5 days.

[0397] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH=6.5), 0.0500 grams of sodium hyaluronate (5 kDa) was added and stirred until it dissolved (4 hours). mg, 0.670 mmol), N,N'-dicyclohexylcarbodiimide (257.3 mg, 1.34 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (100.3 mg, 0.42 mmol) were added. The reaction mixture was stirred in the dark for 20 h, after which it was quenched with hydroxylamine (413.4 mg, 5.95 mmol). The hyaluronic acid product was purified in the dark against deionized water containing decreasing concentrations of salt (NaCl, 0.13 M to 0.0 M) over 5 days. The hyaluronic acid product was filtered (0.22 μm) and lyophilized for 5 days.

[0398] Example A9 Hyaluronic acid modified tetrazine [ka] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH=4.5), 0.5000 grams of sodium hyaluronate (14.8 kDa) was added and stirred until it dissolved. To this, N-hydroxysulfosuccinimide (14.2 mg, 0.0625 mmol), N,N'-dicyclohexylcarbodiimide (125.7 mg, 0.625 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (151.2 mg, 0.625 mmol) were added. The reaction mixture was stirred in the dark for 4 hours, after which it was diluted to 1% (w / w) and then filtered through a 0.45 μm filter. The hyaluronic acid product was then purified by tangential flow filtration (TFF), followed by terminal sterile filtration (0.22 μm) and lyophilization for 3 days. Elemental analysis indicates 40% tetrazine incorporation into the sodium hyaluronate starting material.

[0399] Example A10 Tetrazine-modified alginate gel [ka] To 5 mL of MES buffer (0.1 M MES, 0.3 M NaCl, pH = 6.5), 50 mg of UPLVG alginate (75-200 kDa) was added and stirred until it dissolved (4 h). To this, N-hydroxysulfosuccinimide (34.7 mg, 0.16 mmol), N,N'-dicyclohexylcarbodiimide (61.8 mg, 0.32 mmol), and (4-(6-methyl-1,2,4,5-tetrazin-3-yl)phenyl)methanamine hydrochloride (24.1 mg, 0.10 mmol) were added. The reaction mixture was stirred in the dark for 20 h, after which it was quenched with hydroxylamine (99.3 g, 1.44 mmol). The alginate product was purified in the dark against deionized water containing decreasing concentrations of salt (NaCl, 0.13 M to 0.0 M) over a period of 4 days. The alginate was filtered (0.22 μm) and freeze-dried for 5 days.

[0400] Example C1 In vivo study of antitumor efficacy of drugs in MC-38 subcutaneous allograft model in C57BL / 6 mice 1.Overview BIOMATERIAL1 / PRODRUG1 treatment is a combination therapy of PRODRUG1 (a trans-cyclooctene modified prodrug of doxorubicin) and BIOMATERIAL1 (a tetrazine modified hyaluronic acid biomaterial). PRODRUG1 has attenuated activity and can be administered systemically with minimal risk of spontaneous conversion and exposure to systemic doxorubicin (Dox). PRODRUG1 prodrug is activated only after reacting with BIOMATERIAL1, which is injected into the local site.

[0401] 2. Purpose of the study This study aims to evaluate the in vivo therapeutic efficacy of biomaterial 1 / prodrug 1 treatment in MC38 colon cancer model in immunocompetent C57BL / 6 mice compared to conventional Dox and in combination with TLR agonist (TLRa).

[0402] 3. Study Design Three groups had 5 mice / group, and seven groups had 10 mice / group, for a total of 10 groups.

[0403] 3.1 Treatment Groups and Medication Note: The tumor is approximately 100 mm 3 Treatment was initiated when the mice reached a size of 1. The first day of treatment was designated as "Day 1" as follows.

[0404] [Table 2]

[0405] [Table 3]

[0406] For groups 1–3 (single tumor model), immunocompetent male C57BL / 6 mice were injected with 5 × 10 5, MC38 tumor cells were inoculated SC.

[0407] For groups 4–10 (double tumor model), immunocompetent male C57BL / 6 mice were injected with 5 × 10 5 MC38 tumor cells (large injected tumor) and 1 × 10 5 MC38 tumor cells (small non-injected tumors) were inoculated into each mouse.

[0408] Prior to injection, tumors were grown by suspending MC38 cells in 0.1 mL of DMEM medium mixed with 50% Matrigel. The average tumor volume of large tumors was approximately 100 mm. 3 Once the tumor reached a mass of 100 mg / kg, animals were randomly divided into 10 treatment groups, with 5–10 mice per group, based on body weight and tumor volume. All groups received peritumoral biomaterial injections (100 μL / mouse) near the large tumor (hereafter, injected tumor). One hour later, groups 1, 4, and 10 were IV-administered with saline control (QD×5 days); groups 2, 8, and 9 were IV-administered with Prodrug 1 (PRODRUG1) prodrug (28.6 mg / kg Dox Eq ​​QD×5 days; cumulative dose 143 mg / kg Dox Group 5 received IV Dox HCl control (MTD; 8.1 mg / kg Q4D x 3 doses; cumulative dose 24.3 mg / kg). Tumor volumes of both tumors were measured three times weekly in two dimensions using calipers and volumes were measured in mm 3 Eight groups were used to evaluate inhibition of tumor growth, and groups 9 and 10 (prodrug 1, saline n=10 / group) were used to evaluate inhibition of tumor growth. Immune cell infiltration was determined by flow cytometry. Groups 3, 6, and 7 (n=10) were used to test Dox+TLR9 agonist (SL-01) and prodrug 1+TLR9 agonist. TLR9 agonist was administered as an intratumoral injection into the primary tumor after the last prodrug 1 or Dox dose of 25 μg per mouse. For complete responder mice, 5×10 per mouse 5Tumor rechallenge was performed with 1000 cells. Flow cytometry analysis was performed on animal groups 9 and 10 (prodrug 1, saline n=10 / group) to determine immune cell infiltration by flow cytometry. Tumor harvesting was performed on the second and fourth subgroups after 1 and 2 weeks, respectively. After RBC lysis and Fc blocking, tumor-derived cells were analyzed for cell surface (CD45, CD3, CD4, CD8, CD25, PD-1) or intracellular (FoxP3) markers by flow cytometry. Cells were also labeled with Fixable Viability Stain to distinguish non-viable from viable cells. CD45 is a cell surface (transmembrane) molecule available on most cells of hematopoietic origin (i.e., blood-derived). It is used to distinguish infiltrating cells from native cells in MC38 tumors (a colon carcinoma lineage that lacks CD45). CD3 is a pan-T cell marker, while CD4 and CD8 are T helper cell markers (T H FoxP3 is an intracellular marker present on CD4+CD25+ cells and is typically expressed on T regS PD-1 is identified as the most common type of PD-1. PD-1 is an immune checkpoint protein and a programmed death receptor present on cells, generally T cells. When bound by its cognate ligand, PD-1 can trigger apoptosis of antigen-specific (CD4+ or CD8+) T cells.

[0409] [Table 4]

[0410] 3.2 Retreatment Groups and Dosing Retreatment was performed 38 days post-inoculation (dpi).

[0411] Group 2 - All remaining mice (n=8) 1. One peritumoral injection (through all five poles) of 100 μL of biomaterial 1 2.1 hours later, 5 doses of prodrug 1 at a daily dose of 16.6 mg / kg / day IV (lower dose than before)

[0412] Group 3 - all remaining mice (n=10) Group 3 is divided into two subgroups.

[0413] Each subgroup contained n=5: n=3 had tumors >100 mm 3 and n=2 had tumors <100 mm 3 had.

[0414] One subgroup was treated with the combination of biomaterial 1 + TLRa (peritumoral) and saline alone IV.

[0415] The second subgroup was treated with the combined administration of biomaterial 1 + TLRa (peritumoral) and prodrug 1 administered IV five times a day.

[0416] Combined administration of biomaterial 1 + TLRa 1. 100 μL of biomaterial 1 was mixed with 25 μg of TLRa (for one mouse; a lower amount of TLRa than before) 2. One peritumoral injection of biomaterial 1 mixed with TLRa 3.1 h later, 5 doses of prodrug 1 at a daily dose of 16.6 mg / kg / day IV

[0417] 4.2 Test Substance 4.2.1 Biomaterials 1 having about 10-15 kD MW and about 30% modification, [ka] Tetrazine-modified sodium hyaluronate modified as follows: Appearance: Pink liquid in a syringe Storage: -20℃ Formulation: Not required

[0418] 4.2.2 Prodrug 1 [ka] Appearance: Dry red powder Storage: -20℃ Formulation: Dissolve prodrug 1 powder in sterile phosphate buffered saline (PBS). Adjust pH with 1M NaOH to pH 7.2. Filter through 0.2 μm membrane under sterile conditions. The formulation is prepared fresh each day prior to IV injection.

[0419] 4.2.3 TLRa Invivogen ODN D-SL01 Appearance: Dry white solid Storage: -20℃ Formulation: 200 μg of TLRa is dissolved in 400 μL of endotoxin-free water. The formulation is prepared fresh each day prior to IT injection.

[0420] 4.3 FACS antibodies and reagents

[0421] [Table 5]

[0422] Mouse Fc Block (#553141), BD Horizon Brilliant Stain Buffer (#563794) and compensation beads (#554825) were obtained from BD. ·Miltenyi Tumor Dissociation Kit(Miltenyibiotec,#130-096-730) RBC Lysis Buffer (BD, #555899) 70μm cell strainer (Miltenyibiotec, #130-098-462)

[0423] 5. Experimental methods and procedures: 5.1 Cell culture MC-38 cells were cultured at 37°C in a 5% CO2 incubator with DMEM supplemented with 10% heat-inactivated fetal bovine serum (FBS). Cells were passaged twice a week. Cells were harvested, counted, passaged, and then inoculated when they reached approximately 70% confluence.

[0424] 5.2 Tumor inoculation and classification 85 animals were enrolled in the efficacy study. Tumor volumes were approximately 100 mm 3 Once a mean volume of 100 μL was reached, animals were randomized based on their tumor size using block randomization in Excel as follows: 5×10 cells suspended in 100 μL PBS mixed with 50% matrix gel were randomized based on tumor size as follows: 5 MC-38 cells were inoculated subcutaneously into the right flank. For groups 4 to 10, 1 × 10 cells suspended in 100 μL PBS mixed with 50% matrix gel were inoculated. 5 MC-38 cells were inoculated subcutaneously into the left flank.

[0425] 5.3 Observations During routine monitoring, animals were checked for any adverse effects of tumor growth and / or treatment on normal behavior, such as effects on mobility, food and water consumption (by observation only), and weight gain / loss (weight was measured twice a week during the pre-treatment phase and daily during the treatment phase), eye / hair matting, and any other abnormal effects including tumor ulceration. If animals reached a weight loss of 10%, the sponsor was notified. Unexpected deaths and observed clinical signs were recorded based on the number of animals in each subset. Animals were not allowed to become moribund.

[0426] 5.4 Tumor Measurements and Endpoints Tumor volumes were measured three times weekly in two dimensions using calipers and the volume was calculated according to the formula: V = 0.5a × b 2 (where a and b are the long and short diameters of the tumor, respectively) 3The tumor volumes were then used to calculate both TC and T / C values. TC was calculated using T as the median time (in days) it took for tumors in the treatment group to reach the planned size and C as the median time (in days) it took for tumors in the control group to reach the same size. T / C values ​​(%) were indicators of antitumor efficacy; T and C were the mean volumes of the treatment and control groups, respectively, on a given day. TC values ​​were calculated according to TV. TC was calculated using T as the median time (in days) it took for tumors in the treatment group to reach the planned size and C as the median time (in days) it took for tumors in the control group to reach the same size.

[0427] 5.5 Flow cytometry analysis (FC / FACS) In the same study, two groups (n=10) of mice bearing dual MC38 tumors were used for tumor immune cell profiling, either biomaterial 1 / prodrug 1 treatment or saline treatment. Each group was further divided into two subgroups of 5 mice each. Tumor harvesting was performed after 1 and 2 weeks for the first and second subgroups, respectively. After red blood cell (RBC) lysis and Fc-blocking, tumor-derived cells were analyzed by flow cytometry for cell surface (CD45, CD3, CD4, CD8, CD25, PD-1) or intracellular (FoxP3) markers. Cells were also labeled with Fixable Viability Stain to distinguish non-viable from viable cells. The percentage of cells in each population of interest was determined (Table 6). CD45 is a cell surface (transmembrane) molecule available on most cells of hematopoietic origin (i.e., blood-derived). It is used to distinguish infiltrating cells from native cells in MC38 tumors (a colon carcinoma lineage that lacks CD45). CD3 is a pan-T cell marker, while CD4 and CD8 are present on helper T cells (TH cells) and cytotoxic T lymphocyte (CTL) subsets, respectively. FoxP3 is an intracellular marker present on CD4+CD25+ cells and is typically associated with regulatory T cells (T regSPD-1 is an immune checkpoint protein and a programmed death receptor present on cells, typically T cells. When bound by its cognate ligand, PD-1 can trigger apoptosis of antigen-specific (CD4+ and CD8+) T cells.

[0428] 5.5.1 Tissue processing For tumor samples, the tumor tissue was homogenized using the Miltenyi Tumor Dissociation kit. Single cell suspensions from tumor samples were RBC lysed, centrifuged to pellet cells, washed with cold PBS, and then the cell pellets were resuspended in staining buffer and stored on ice. The cells were then ready for FACS antibody staining.

[0429] 5.5.2 FACS antibody staining 1. For each sample, add cells (≤10 × 10) to staining buffer in an Eppendorf tube labeled with the correct sample name. 6 cells / tube) were added. Panel 1: Viability test reagents / CD45 / CD3 / CD4 / CD8 / CD25 / PD-1 Panel 2: Viability test / CD45 / CD3 / CD4 / CD25 / Fox3

[0430] 2. For each tissue type (tumor), prepare two separate tubes (see below) and add 10 x 10 cells (≤10 x 10) to the staining buffer in these two tubes. 6 cells / tube) were added. Tube: Colorless Tube: Live-dead only*** Tube: Colorless (fixed / transparent) Tube: Viability test reagent only (fixation / permeabilization) NOTE: These two tubes were used as gating controls for each tissue type. *** A tube containing only viability reagent was also used for viability reagent compensation purposes. Compensation for the eight fluorochrome-conjugated antibodies was prepared separately using compensation beads from BD according to the vendor's manual.

[0431] 3. Prepare FcR blocking solution (100 μL per tube x number of tubes) by diluting FcR blocking antibody 1 / 100 in staining buffer. Note: Because two Brilliant Violet dyes were used in the FACS panel, BD Horizon Brilliant Stain Buffer was added at 5 μL per 100 μL of FcR Blocking Solution.

[0432] 4. Resuspend the cell pellet in 100 μL of FcR blocking solution and incubate for 3 min at RT in the dark.

[0433] 5. For panel 1, FACS antibodies were added to the desired concentration (2 μg / ml for each antibody). Incubated 30 minutes at 4°C in the dark. For panel 2, FACS antibodies (except foxp3) were added to the desired concentration (2 μg / ml for each antibody) and incubated 30 minutes at 4°C in the dark, after which cells were fixed / permeabilized for 40-50 minutes at 4°C before adding foxp3 antibody to the desired concentration (2 μg / ml) over 40 minutes at 4°C in the dark.

[0434] 6.1 ml of staining buffer was added and the cells were washed and pelleted by centrifugation at 350 g for 4 minutes at 4°C.

[0435] 7. Resuspend the cells in "Tube: Colorless" containing 500 μL of staining buffer (no viability reagent added).

[0436] Cells were resuspended in all other tubes containing 500 μL of staining buffer with 8.1 μg / ml of viability reagent.

[0437] 9. After the cells were ready for FACS analysis, they were subjected to analysis using an Attune Nxt Flow Cytometer (ThermoFisher).

[0438] 5.6 Statistical analysis Two-way ANOVA was performed to compare body weight and tumor volume. All data were analyzed using GraphPad Prism 5. In our analysis, p<0.05 was considered statistically significant.

[0439] 5.7 Endpoint Tissue Collection Five mice were sacrificed for Groups 9 and 10, respectively, 1 and 2 weeks after completion of treatment, and tumors were harvested for FACS.

[0440] 6. Results and Discussion 6.1 Weight The results of the weight changes in the tumor-bearing mice for groups 1 to 8 are shown in FIG. 1A and FIG. 1B.

[0441] 6.2 Tumor volume Injected tumor volumes for all treatment groups at various time points are shown in Figures 2A and 2B for Groups 1-8. Injected tumor volumes for all treatment groups analyzed are shown in Figures 3A and 3B for Groups 1-8. Non-injected tumor volumes for all treatment groups at various time points are shown in Figure 4 for Groups 4-8. Non-injected tumor volumes for all treatment groups analyzed are shown in Figure 5 for Groups 4-8.

[0442] 6.3 Tumor growth inhibition effect Treatment with biomaterial 1 / prodrug 1 therapeutics resulted in significantly improved antitumor responses (p<0.05) and overall survival (p<0.001) compared to those seen with conventional Dox treatment (Figures 6A-6B and 6D). Furthermore, the majority of non-injected tumors in the biomaterial 1 / prodrug 1 treatment group showed sustained antitumor responses, whereas all non-injected tumors in the conventional Dox treatment group showed progressive growth (Figure 6C).

[0443] The tumor growth inhibitory effects are summarized in Tables 3, 4, and 5.

[0444] [Table 6]

[0445] [Table 7]

[0446] [Table 8]

[0447] Immunocompetent C57BL / 6 mice were inoculated with MC38 tumors. All tumor cells were implanted on day 0. Treatment began on day 7 with local injection of biomaterial into the "injected" tumors, followed by systemic therapy. Large (injected) tumors were 5×10 5 Small (non-injected) tumors were started with 1 × 10 5 Tumor growth curves show the mean ± SEM; data points without error bars occurred when the standard error was smaller than the sign used to represent the treatment condition. Curves were discontinued when one or more mice in the group died or the tumor volume exceeded 2000 mm 3 and stopped after being sacrificed.

[0448] *, **Statistical significance of tumor growth curves was determined by unpaired t-test with Welch's correction for each day. Saline (Group 4) and Dox HCl (Group 5) treatments were not significantly different on any day. Biomaterial 1 / Prodrug 1 (Group 8) treatment was significantly different from saline, Dox HCl, or both saline and Dox HCl treatments on the days indicated by the asterisk and brackets.

[0449] ***Statistical significance for survival times was determined by log-rank (Mantel-Cox) test; biomaterial 1 / prodrug 1 treatment was significantly different from Dox HCl or saline, whereas Dox HCl and saline were not significantly different from each other.

[0450] Immunocompetent C57BL / 6 mice were inoculated with MC38 tumors. All tumor cells were inoculated on day 0. Treatment was started on day 7 with local injection of biomaterial into the "injected" tumors, followed by systemic therapy. Large (injected) tumors were 5×10 5 Small (non-injected) tumors were started with 1 × 10 5 Tumor growth of individual non-injected tumors is expressed as a percentage of the initial volume of each tumor (measurements started 12 days after inoculation).

[0451] 6.5 Flow cytometry analysis Tumor samples were taken for immune profiling after 1 or 2 weeks of treatment. Table 6 shows the immune cell populations and the corresponding markers used for detection. Figures 8 and 9 show the quantification of immune cell frequency and phenotype in tumor samples after 2 weeks.

[0452] One week after completion of treatment, no differences were observed between the treatment and saline groups. Two weeks after completion of treatment, significant differences in T cell profiles were observed between the Biomaterial 1 / Prodrug 1 treated and saline groups for both injected and non-injected tumors. There was an increase in the overall % of CD45+CD3+ cells in Biomaterial 1 / Prodrug 1 treated mice compared to that shown in saline treated mice for both injected ("Injected") (Figure 8) and non-injected ("Non-Injected") (Figure 9) tumors. This indicated an increase in total tumor infiltrating lymphocytes (TILs) in Biomaterial 1 / Prodrug 1 treated mice. Among these cells, the percentages of C8+ and CD4+ cells were significantly higher in the injected tumors receiving Biomaterial 1 / Prodrug 1 treatment compared to saline (Figure 8). For non-injected tumors, only the percentage of CD4+ cells was significantly higher in the Biomaterial 1 / Prodrug 1 treated group compared to the saline group (Figure 9). In the case of non-injected tumors (FIG. 9), no CD4+CD25+FoxP3+ cells were observed in the biomaterial 1 / prodrug 1 treatment group.

[0453] Taken together, these results indicate an overall increase in helper T cells and CTL effectors, as well as an increase in T cells infiltrating tumors in the biomaterial 1 / prodrug 1 treatment group. reg The cell percentage of CD8+ cells in the non-injected tumors (FIG. 9) was found to be higher, whereas the percentage of FoxP3+ cells in the injected tumors (FIG. 8) was lower in the saline group, and more animals per group would have been needed to achieve statistical significance.

[0454] Interestingly, there was a higher percentage of PD-1+CD4+ T cells in the injected tumors of the biomaterial 1 / prodrug 1 treatment group (Figure 8) compared to that shown in the saline group. Although the elevation of PD-1 indicates T cell exhaustion, the true functional significance of these cells needs to be further explored. This difference was not observed in the non-injected tumors (Figure 9). This finding also provides a solid basis for the application of the combination therapy of biomaterial 1 / prodrug 1 and anti-PD-1 checkpoint inhibitors in future studies. Furthermore, there was no difference in PD-1 expression in CD8+ cells in injected or non-injected tumors in saline or biomaterial 1 / prodrug 1 treatment groups.

[0455] Taken together, the flow cytometry data from this study demonstrated that Biomaterial 1 / Prodrug 1 treatment has the capacity for immune activation and increases the total number of TILs in both injected and non-injected tumors. These effects were present after 2 weeks but not after 1 week of treatment, suggesting a time response. Solid tumors are reg This study demonstrated that Biomaterial 1 / Prodrug 1 treatment suppresses T cell proliferation in non-injected tumors. reg These results suggest that this may reduce the number of PD-1 cells and may have potential benefit when combined with anti-PD-1 antibodies.

[0456] [Table 9]

[0457] 7. Results of the re-challenge test Apart from the double tumor group, the study also examined tumor reimplantation in mice inoculated with only one MC38 tumor. On day 38, all animals (Table 1) (n=8) in the G2 group were treated with a second cycle of biomaterial 1 / prodrug 1 (injection of 100 μL of intratumoral biomaterial 1 followed by five daily doses (one dose per day) of prodrug 1 - this time 11.9 mg / kg / dose of Dox Eq (59.3 mg / kg / cycle of Dox Eq).

[0458] On day 70, one mouse from group G2 with a tendency for complete response (Figure 10A) was inoculated SC in the left flank with 5 x 10 5 A control group of five naive mice was rechallenged with 5 × 10 MC38 tumor cells on the same day. 5 MC38 tumor cells were inoculated into the treated and untreated animals. Tumor growth curves for treated and untreated animals are shown in Figure 10B.

[0459] In addition, mice from group G3 (Table 1) (n=5) and mice from dual tumor groups G6, G7, and G8 (n=10) were rechallenged. Tumors grew rapidly in all naive mice, whereas tumor growth was suppressed in Biomaterial 1 / Prodrug 1 pretreated mice, suggesting that Biomaterial 1 / Prodrug 1 treatment may trigger an antitumor memory immune response.

[0460] 8. Summary and Conclusion This study evaluates the therapeutic efficacy of prodrug 1, prodrug 1 + TLR agonist, Dox, and Dox + TLRa in a subcutaneous MC-38 tumor model.

[0461] Treatment with prodrug 1, prodrug 1 + TLR agonist resulted in significant antitumor activity compared to control group G1, P<0.001, P<0.001, respectively. Treatment of injected tumors with prodrug 1, prodrug 1 + TLR agonist, Dox, and Dox + TLR agonist resulted in significant antitumor activity compared to control group G4, P<0.01, P<0.001, P<0.001, respectively. Treatment of non-injected tumors with prodrug 1, prodrug 1 + TLR agonist, Dox, and Dox + TLR agonist resulted in significant antitumor activity compared to G4, P<0.001, P<0.001, P<0.001, respectively. All treatments were well tolerated in MC-38 tumor-bearing C57BL / 6 mice.

[0462] Taken together, the flow cytometry data from this study demonstrated that Biomaterial 1 / Prodrug 1 treatment has the capacity for immune activation and increases the total number of TILs in both injected and non-injected tumors. These effects were present after 2 weeks but not after 1 week of treatment, suggesting a time response. Solid tumors are reg This study demonstrated that Biomaterial 1 / Prodrug 1 treatment suppresses T cell proliferation in non-injected tumors. reg These results suggest that this may reduce the number of PD-1 cells and may have potential benefit when combined with anti-PD-1 antibodies.

[0463] Sixteen mice were rechallenged with MC-38 cells and compared to the vehicle group at the end of the reimplantation study, all mice's tumors still showed significant changes. In comparison, all 10 of the 10 naive mice implanted with MC-38 cells had tumors that grew normally. This data suggests that the treatment can induce an anti-tumor immune memory response.

[0464] In conclusion, biomaterial 1 / prodrug 1 treatment showed tumor growth inhibition and improved overall survival compared to conventional Dox treatment. In addition, it showed complete remission in 10% of treated mice, followed by a sustained antitumor response upon rechallenge. Furthermore, biomaterial 1 / prodrug 1 treatment induced immune activation and resulted in an increase in total TILs 2 weeks after the start of therapy. Overall, biomaterial 1 / prodrug 1 treatment demonstrated sustained anticancer and immunomodulatory effects in both injected and non-injected tumors, suggesting that it may be useful for the treatment of localized tumors and metastatic disease.

[0465] For completeness, the various aspects of the invention are set out in the following numbered clauses: Clause A1. A compound of formula (IA) or a pharma- ceutically acceptable salt thereof [ka] (In the formula, R 1a Each occurrence of represents hydrogen, C 1~4 Alkyl, and C 1~4 independently selected from the group consisting of haloalkyl; R 1b Each occurrence of represents hydrogen, C 1~4 Alkyl, C 1~4 Haloalkyl, C(O)OH, C(O)OC 1~4 Alkyl, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )CHR 1e C(O)OC 1~4 Alkyl, C(O)N(R 1c )-C 1~6 Alkylene-COH and C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 independently selected from the group consisting of alkyl; R 1c Each occurrence is independently hydrogen or C 1~4 is alkyl; R1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; D, at each occurrence, is independently a payload selected from the group consisting of a toll-like receptor (TLR) agonist and a stimulator of interferon genes (STING) agonist; L 1 is, independently at each occurrence, a linker; m is independently at each occurrence 1, 2, or 3; Each occurrence of p is independently 0, 1, or 2.

[0466] Clause A2.R 1a is hydrogen; R 1b is hydrogen, or a pharma- ceutically acceptable salt thereof.

[0467] Article A3.R 1a is C 1~4 is alkyl; R 1b are C(O)OH and C(O)OC. 1~4 Alkyl, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )CHR 1e C(O)OC 1~4 Alkyl, C(O)N(R 1c )-C 1~6 Alkylene-COH and C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 The compound of clause A1, wherein the compound is selected from the group consisting of alkyl, or a pharma- ceutically acceptable salt thereof.

[0468] Article A4.R 1b are C(O)OH, C(O)N(R 1c )CHR 1e COH and C(O)N(R 1c)CH2CO2H, or a pharma- ceutically acceptable salt thereof.

[0469] Article A5.R 1e is -CH2CO2H, -CH2CH2CO2H, -CH2CONH2, -CH2CH2CONH2, -CH2OH, or -CH(CH3)OH, or a pharma- ceutically acceptable salt thereof.

[0470] Article A6.R 1e -C 1~4 A compound according to clause A3 or A4, which is alkylene-CO2H, or a pharma- ceutically acceptable salt thereof.

[0471] Article A7.R 1e is -CH2CO2H, or a pharma- ceutically acceptable salt thereof.

[0472] Article A8.R 1a A compound according to any of clauses A3 to A7, or a pharma- ceutically acceptable salt thereof, wherein:

[0473] Clause A9.R 1c A compound according to any of clauses A3 to A8, wherein is hydrogen, or a pharma- ceutically acceptable salt thereof.

[0474] Article A10.L 1 teeth, [ka] or -O-; L 3 is a bond or C 1~6 is alkylene; L 4 is a bond, -NHN:, -N(R 10 )-C 2~6 Alkylene-N(R 11 )-, -N(R 12 )-C 2~3 Alkylene-N(R 13 )C(O)-, -N(R 10 )-C1~6 Alkylene-C(O)NHN: -NHNHC(O)C 1~6 Alkylene -C(O)NHN: -CH(NHC(O)R 14 ) C 1~4 Alkylene-SSC 1~4 Alkylene -OC(O)-, -NHNHC(O)CH(NHC(O)R 15 )CH2C(O)-, -C 1~6 Alkylene-CH(G x )OC(O)-, [ka] and; R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , and R 19 are each independently hydrogen or C 1~4 is alkyl; R 16 is hydrogen, C 1~4 Alkyl, -C 1~4 Alkylene -OH, -C 1~4 Alkylene-OC 1~4 Alkyl, -C 1~4 Alkylene-COH or -C 1~4 alkylene-CONH2; R 17 is independently at each occurrence hydrogen or -CH2OC(O)-; G x Optionally, halogen, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 The compound of any of clauses A1-A9, or a pharma- ceutically acceptable salt thereof, wherein phenyl is substituted with one to five substituents independently selected from the group consisting of alkoxy, cyano, and nitro.

[0475] Clause A11.m is a compound according to any of clauses A1 to A10, which is 1, or a pharma- ceutically acceptable salt thereof.

[0476] Article A12. [ka] teeth, [ka] [ka] and R 18 is independently at each occurrence hydrogen or -CH2OC(O)NHD'; R D is a hydrogen or C on the nitrogen atom of the payload 1~4 is alkyl; D' is a payload moiety, the compound of clause A11, or a pharma- ceutically acceptable salt thereof.

[0477] Clause A13. A compound according to any of clauses A1 to A12, wherein p is 0, or a pharma- ceutically acceptable salt thereof.

[0478] Clause A14.m is a compound of clause A13 which is 2 or 3, or a pharma- ceutically acceptable salt thereof.

[0479] Article A15. [ka] teeth, [ka] or a pharma- ceutically acceptable salt thereof.

[0480] Clause A16. Payload D is: [ka] A compound according to any one of clauses A1 to A15, selected from the group consisting of:

[0481] Article A17 below: [ka] [ka] [ka] or a pharma- ceutically acceptable salt thereof.

[0482] Clause A18. A pharmaceutical composition comprising a compound according to any one of clauses A1 to A17, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0483] Clause A19. A method for treating or preventing a disease state or disorder or enhancing or inducing an immune response, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of clauses A1-A17, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to clause A18, and a therapeutic support composition, the therapeutic support composition comprising a biocompatible support and a compound of the formula [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2 is selected from the group consisting of NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S) R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; t is 0, 1, 3, or 4. The method comprises the steps of:

[0484] Clause A20. The method of clause A19, wherein the tetrazine-containing group is linked or directly bonded to the hyaluronic acid biocompatible support.

[0485] Clause A21. The therapeutic supportive composition comprises a compound of formula (II) [ka] (In the formula, G 2 teeth, [ka] and; R 22 is a linker of 1 to 100 linking atoms. The method of clause A20, comprising a substituted hyaluronic acid unit of

[0486] Article A22.G 2 teeth, [ka] That is, the method of clause A21.

[0487] Article A23.G 2 teeth, [ka] and; R 20 is hydrogen or C 1~4 The method of clause A21, wherein the alkyl group is alkyl.

[0488] Clause A24. The method of any of clauses A19-A23, wherein said method is a method for treating or preventing cancer.

[0489] Clause A25. The method of clause A24, wherein said cancer is melanoma, renal cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer.

[0490] Clause A26. The method of clause A24 or A25, wherein said cancer is a solid tumor.

[0491] Clause A27. The method of clause A24 or A25, wherein said cancer is a soft tissue sarcoma.

[0492] Clause A28. The method of clause A27, wherein said soft tissue sarcoma is fibrosarcoma, rhabdomyosarcoma, or Ewing's sarcoma.

[0493] Clause A29. The method of any of clauses A19 to A23, wherein said method is a method of enhancing or inducing an immune response.

[0494] Clause A30. The method of clause A29, wherein said immune response is an increase in one or more of leukocytes, lymphocytes, monocytes, and eosinophils.

[0495] Clause A31. A therapeutically effective amount of: an anticancer drug, an immune checkpoint inhibitor, or a compound of formula (IB), or a pharma- ceutically acceptable salt thereof. [ka] (In the formula, D 1 Each occurrence is independently classified as an anticancer drug payload, a microbial immunosuppressant payload, a restenosis inhibitor payload, an antibiotic payload, an antifungal payload, an antiviral payload, a payload selected from an anti-inflammatory / anti-arthritic payload, a corticosteroid payload, and an immunosuppressant payload; R 1a , R 1b , L 1 , p, and m are as defined in any one of claims 1 to 11. The method of any of clauses A19-A30, further comprising administering another additional therapeutic agent selected from the group consisting of:

[0496] Clause A32.p is 0; m is 1;-L 1 -teeth, [ka] That is, the method of clause A31.

[0497] Clause A33. The method of clause A31 or A32, wherein said anticancer drug is doxorubicin.

[0498] Clause A34. A kit comprising a compound according to any one of clauses A1 to A17, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to clause 18, and instructions for use thereof.

[0499] Clause A35. The kit of clause A34, further comprising a therapeutic support composition as defined in any of clauses A19-A23.

[0500] Clause A36. A kit according to clause A34 or A35, further comprising a compound of formula (IB) as defined in any of clauses A31 to A33.

[0501] Clause B1. A method of treating cancer or enhancing or inducing an immune response comprising administering to a subject in need thereof: a) a therapeutically effective amount of a compound of formula (II-A) or a pharma- ceutically acceptable salt thereof; [ka] (In the formula, R 1A Each time it occurs, C 1~4 Alkyl, C 1~4 Haloalkyl and C 1~4 independently selected from the group consisting of alkoxy; R 1B Each time it occurs, G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1~6 Alkylene-COH, -N(R 1f )-C 2~4 Alkylene-( N(C 1~4 Alkylene-COH)-C 2~4 Alkylene) n -N(C 1~4Alkylene -COH)2, -N(R 1c )CHR 1e C(O)OC 1~6 Alkyl, -N(R 1c )-C 1~6 Alkylene-C(O)OC 1~6 Alkyl, and -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-C(O)OC 1~6 Alkyl)-C 2~4 Alkylene) n -N(C 1~4 Alkylene-C(O)OC 1~6 alkyl)2; R 1c and R 1d Each occurrence is independently hydrogen or C 1~4 is alkyl; R 1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; R 1f Each occurrence independently represents hydrogen, C 1~6 Alkyl or -C 1~4 alkylene-COH; D 1 is, each occurrence independently, an anticancer drug payload; L 1 is, independently at each occurrence, a linker; L 2 Each occurrence independently represents -C(O)- and C 1~3 alkylene; G 1 is independently at each occurrence an optionally substituted heterocyclyl; m is 1, 2, or 3; n is independently at each occurrence 0, 1, 2, or 3; p, at each occurrence, is independently 0, 1, or 2; b) a support and a compound of formula [ka] (In the formula, R 20 is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2NR'R'', C(=O)O-R', C(=O)S-R', C(=S) is selected from the group consisting of O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R'', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1- C6 haloalkyl; t is 0, 1, 3, or 4. a therapeutic support composition comprising a tetrazine-containing group of the formula (wherein the tetrazine-containing group is linked or directly bonded to the support); c) a therapeutically effective amount of one or more immunomodulatory agents, or pharma- ceutically acceptable salts thereof; The method of claim 1, further comprising administering

[0502] Clause B2. The method of clause B1, wherein said method is a method of enhancing or inducing an immune response, and administering a), b), and c) enhances or induces an immune response against cancer in the subject.

[0503] Clause B3. The method of clause B1 or B2, wherein said immune response is an increase or decrease in one or more of innate and adaptive immune cells, including but not limited to, leukocytes, lymphocytes, monocytes, eosinophils, and antibodies.

[0504] Clause B4. The method of clause B1, wherein said method is a method of treating cancer.

[0505] Clause B5. The method of any of clauses B1-B4, wherein said cancer is melanoma, renal cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, rhabdomyosarcoma, colon cancer or pancreatic cancer.

[0506] Clause B6. The method of any of clauses B1-B5, wherein said cancer is a solid tumor.

[0507] Clause B7. The method of any of clauses B1-B5, wherein said cancer is soft tissue sarcoma.

[0508] Clause B8. The method of clause B7, wherein said soft tissue sarcoma is fibrosarcoma, rhabdomyosarcoma, or Ewing's sarcoma.

[0509] Clause B9. The method of any of clauses B1-B5, wherein said cancer is diffuse pontine glioma.

[0510] Clause B10. The method of any of clauses B1-B9, further comprising administering a therapeutically effective amount of an immune checkpoint inhibitor.

[0511] Clause B11. The method of any of clauses B1-B10, wherein said immunomodulatory agent is a toll-like receptor (TLR) agonist.

[0512] Clause B12. The method of clause B11, wherein said toll-like receptor (TLR) agonist is Bacillus Calmette-Guerin (BCG), lipopolysaccharide, peptidoglycan, polyriboinosinic-polyribocytidylic acid (Poly I:C), imiquimod, Coley's toxin, polyadenylic-polyuridylic acid (Poly A:U), monophosphoryl lipid A, single-stranded and double-stranded RNA, or CpG oligodeoxynucleotide (ODN).

[0513] Clause B13. The method of any of clauses B1-B10, wherein said immunomodulatory agent is a stimulator of interferon genes (STING) agonist.

[0514] Clause B14. The method of any of clauses B1-B10, wherein said immunomodulatory agent is a cytokine, a cytokine inhibitor, a cytokine receptor agonist, or a cytokine receptor antagonist.

[0515] Clause B15. The immunomodulatory agent is selected from the group consisting of chemokines, chemokine inhibitors, chemokine receptor antagonists, and the like. The method of any of clauses B1-B10, wherein the compound is a vasopressin receptor antagonist, a vasopressin receptor antagonist, or a chemokine receptor antagonist.

[0516] Clause B16. The method of any of clauses B1-B15, wherein a), b) and c) are administered simultaneously, separately or sequentially, in any order.

[0517] Clause B17. The method of any of clauses B1-B16, wherein the immunomodulatory agent is administered simultaneously with the therapeutic support composition.

[0518] Clause B18. The method of clause B16 or B17, wherein the simultaneous administration is by co-injection, co-implantation, or co-formulation.

[0519] Article B19.R 1B is G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, and -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1e is -CH2CO2H, -CH2CH2CO2H, -CH2CONH2, -CH2CH2CONH2, -CH2OH, or -CH(CH3)OH; R 1f The process of any of clauses B1-B18, wherein is hydrogen or CH2CO2H.

[0520] Article B20.R 1A is C 1~4 is alkyl; R 1B is G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )CH2CO2H, and -N(R 1f )-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2; R 1e -C 1~4 alkylene-COH; R 1f is hydrogen or C 1~4 alkylene-COH; G 1is a 4-8 membered monocyclic heterocyclyl containing a first nitrogen and optionally one additional heteroatom selected from nitrogen, oxygen, and sulfur; G 1 is attached to the first nitrogen, and optionally, C 1~4 Alkyl, C 1~4 Haloalkyl, halo, cyano, OH, -OC 1~4 substituted with 1 to 4 substituents independently selected from the group consisting of alkyl, aryl, aryloxy ... The method of any of clauses B1-B18, wherein n is 0, 1, or 2.

[0521] Article B21.R 1A is CH3; R 1e is -CH2CO2H; R 1f is hydrogen or CH2CO2H; G 1 is attached via a ring nitrogen atom, and optionally 1~4 Alkyl, C 1~4 Haloalkyl, halo, cyano, OH, -OC 1~4 The process of clause B20, wherein the is piperazinyl, morpholinyl, piperidinyl, azepanyl, or pyrrolidinyl substituted with 1 to 4 substituents independently selected from the group consisting of alkyl, and oxo.

[0522] Article B22.L 2 The method of any of clauses B1 to B21, wherein -C(O)-.

[0523] Article B23.R 1B are OH, N(H)CH2CO2H, -N(H)CHR 1e CO2H, -N(H)-CH2CH2-(N(CH2CO2H)CH2CH2) n -N(CH2CO2H)2, and -N(CH2CO2H)-CH2CH2-N(CH2CO2H)2; R 1e is -CH2CO2H, The method of clause B22.

[0524] Article B24.L1 teeth, [ka] or -O-; L 3 is a bond or C 1~6 is alkylene; L 4 is a bond, -NHN:, -N(R 10 )-C 2~6 Alkylene-N(R 11 )-, -N(R 12 )-C 2~3 Alkylene-N(R 13 )C(O)-, -N(R 10 )-C 1~6 Alkylene-C(O)NHN: -NHNHC(O)C 1~6 Alkylene -C(O)NHN: -CH(NHC(O)R 14 ) C 1~4 Alkylene-SSC 1~4 Alkylene -OC(O)-, -NHNHC(O)CH(NHC(O)R 15 )CH2C(O)-, -C 1~6 Alkylene-CH(G x )OC(O)-, [ka] and; R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , and R 19 are each independently hydrogen or C 1~4 is alkyl; R 16 is hydrogen, C 1~4 Alkyl, -C 1~4 Alkylene -OH, -C 1~4 Alkylene-OC 1~4 Alkyl, -C 1~4 Alkylene-COH or -C 1~4 alkylene-CONH2; R17 is independently at each occurrence hydrogen or -CH2OC(O)-; G x Optionally, halogen, C 1~4 Alkyl, C 1~4 Haloalkyl, C 1~4 The process of any of clauses B1-B23, wherein the phenyl is substituted with 1 to 5 substituents independently selected from the group consisting of alkoxy, cyano, and nitro.

[0525] Clause B25.m is 1, any one of clauses B1 to B24.

[0526] Article B26. [ka] teeth, [ka] [ka] and; R 18 Each occurrence is independently hydrogen or -CH2OC(O)NHD 1a and; R D is a hydrogen or C on the nitrogen atom of the payload 1~4 is alkyl; D 1a The method of clause B25, wherein:

[0527] The method according to any one of clauses B1 to B26, wherein clause B27.p is 0.

[0528] Clause B28.m is 2 or 3, the method of clause B27.

[0529] Article B29. [ka] teeth, [ka] That is, the method of clause B28.

[0530] Clause B30. The therapeutic support composition comprises a compound of formula (II): [ka] (In the formula, G 2 teeth, [ka] and; R 22 is a linker of 1 to 100 linking atoms. The method of any of clauses B1-B29, comprising a substituted hyaluronic acid unit of

[0531] Article B31.G 2 teeth, [ka] This is the method of clause B30.

[0532] Article B32.G 2 teeth, [ka] and; R 20 is hydrogen or C 1~4 The method of clause B31, wherein the alkyl group is alkyl.

[0533] Article B33 below: a) a compound of formula (IA) as defined in any of clauses B1 or B19-B29, or a pharma- ceutically acceptable salt or composition thereof; b) one or more immunomodulatory agents, or pharma- ceutically acceptable salts or compositions thereof; c) Instructions for use; Including the kit.

[0534] Clause B34. The kit of clause B33 further comprising a therapeutic support composition according to any of clauses B1 or B30-B32.

[0535] Article B35 et seq.: a) a therapeutic support composition according to any of clauses B1 or B30-B32; b) one or more immunomodulatory agents, or pharma- ceutically acceptable salts or compositions thereof; c) Instructions for use; Including the kit.

[0536] Article B36. Below: a) a compound of formula (IA) according to any of clauses B1 or B19 to B29, or and its salts which are acceptable to; b) one or more immunomodulatory agents, or pharma- ceutically acceptable salts thereof; c) a pharma- ceutically acceptable carrier; and 23. A pharmaceutical composition comprising:

[0537] Article B37 below: a) a therapeutic support composition according to any of clauses B1 or B30-B32; b) one or more immunomodulatory agents, or pharma- ceutically acceptable salts thereof; c) a pharma- ceutically acceptable carrier; and 23. A pharmaceutical composition comprising:

[0538] Clause B38. The method, composition, or kit of any of clauses A1-A36 and B1-B37, wherein the support is a polysaccharide hydrogel, alginate, agarose, cellulose, hyaluronic acid, chitosan, chitin, chondroitin sulfate, heparan sulfate, heparin, gelatin, collagen, a polymer matrix, metal, ceramic, or plastic, each of which may be optionally modified.

[0539] Clause C1. A method of treating cancer comprising: a) administering to a subject in need thereof a therapeutically effective amount of a compound of formula (II-A), or a pharma- ceutically acceptable salt thereof. [ka] (In the formula, R 1A is C 1~4 Alkyl, C 1~4 Haloalkyl and C 1~4 alkoxy; R 1B is G 1 , OH, -NR 1c -C 1~4 Alkylene-G 1 , -NR 1c -C 1~4 Alkylene-N(R 1d )2, -N(R 1c )CHR 1e CO2H, -N(R 1c )-C 1~6 Alkylene-COH, -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-COH)-C 2~4 Alkylene) n -N(C 1~4 Alkylene -COH)2, -N(R 1c )CHR 1e C(O)OC 1~6 Alkyl, -N(R 1c )-C 1~6 Alkylene-C(O)OC 1~6 Alkyl, and -N(R 1f )-C 2~4 Alkylene-(N(C 1~4 Alkylene-C(O)OC 1~6 Alkyl)-C 2~4 Alkylene) n -N(C 1~4 Alkylene-C(O)OC 1~6 alkyl)2; R 1c and R 1dEach occurrence is independently hydrogen or C 1~4 is alkyl; R 1e -C 1~4 Alkylene-COH, -C 1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; R 1f is hydrogen, C 1~6 Alkyl or -C 1~4 alkylene-COH; D 1 is, each occurrence independently, the payload; -L 1 - is a linker; -L 2 - is -C(O)- and C 1~3 alkylene; G 1 is an optionally substituted heterocyclyl; m is 1, 2, or 3; n is 0, 1, 2, or 3; p is 0, 1, or 2; and b) a support and a compound of formula [ka] (In the formula, R 20is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF3, CF2-R', NO2, OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O)2R''', S(=O)2 is selected from the group consisting of NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S) R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R 30 is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R a , R 31a and R 31b are each independently hydrogen, C1-C6 alkyl, or C1-C6 haloalkyl; t is 0, 1, 3, or 4. locally administering to the subject a therapeutic support composition comprising a tetrazine-containing group of the formula: Including, wherein the subject has a second tumor, and administration of a) and administration of b) inhibits growth of the second tumor.

[0540] Clause C2. A method of enhancing or eliciting an immune response to a second tumor in a subject, comprising: a) administering to a subject a compound of formula (IA), or a pharma- ceutically acceptable salt thereof. and b) locally administering a therapeutic support composition to a first tumor in a subject. Including, wherein the compound of formula (IA) and the therapeutic support composition are as defined in clause C1; The administration of a) and the administration of b) enhances or induces an immune response against a second tumor.

[0541] Clause C3. The method of clause C1 or C2, wherein the therapeutic support composition is not administered locally to the second tumor.

[0542] Clause C4. A method of inhibiting tumor metastasis in a subject at risk of tumor metastasis, comprising: a) administering to a subject a compound of formula (IA), or a pharma- ceutically acceptable salt thereof; and b) locally administering a therapeutic support composition to a first tumor in a subject; A method wherein the compound of formula (IA) and the therapeutic support composition are as defined in clause C1.

[0543] The method of clause C4, wherein administering clause C5.a) and administering clause b) enhances or induces an immune response that inhibits metastasis.

[0544] Clause C6. The method of clause C5 or C6, wherein inhibiting tumor metastasis comprises inhibiting the development of a second tumor in the subject.

[0545] Clause C7. The method of any of clauses C4-C6, further comprising the step of identifying a subject at risk for tumor metastasis.

[0546] Clause C8. The method of any of clauses C4-C7, further comprising the step of selecting a subject at risk for tumor metastasis.

[0547] Clause C9. The method of any of clauses C4-C8, wherein the subject at risk of metastasis is afflicted with a first tumor that is considered to be a stage II-III or higher solid tumor or a high-grade tumor.

[0548] Clause C10. The method of any of clauses C4-C9, wherein the first tumor cell is isolated from the first tumor.

[0549] Clause C11. The method of clause C10, wherein the first tumor cell is present in tissue surrounding the first tumor, present in a tumor cell-platelet aggregate, present in the subject's systemic circulation, and / or present at a second tumor site in the subject.

[0550] Clause C12. The method of any of clauses C1-C11, wherein said subject exhibits a biomarker of tumor metastasis.

[0551] Clause C13. The method of clause C12, wherein said biomarker is one or more of CCR7, CXCR4, E-cadherin, EpCAM, VCAM1, integrin-alpha10, N-cadherin, vimentin, and fibronectin.

[0552] Clause C14. The composition of clauses C1 to C13, further comprising a therapeutically effective amount of one or more immunomodulatory agents. Either way.

[0553] Clause C15. The method of clause C14, wherein the one or more immunomodulatory agents are one or more of an immune checkpoint inhibitor, a toll-like receptor (TLR) agonist, a stimulator of interferon genes (STING) agonist, a cytokine, a cytokine inhibitor, a cytokine receptor agonist, a cytokine receptor antagonist, a chemokine, a chemokine inhibitor, a chemokine receptor agonist, or a chemokine receptor antagonist.

[0554] Clause C16. The method of clause C14 or C15, wherein the one or more immunomodulatory agents comprise one or more TLR agonists selected from the group consisting of Bacillus Calmette-Guerin (BCG), lipopolysaccharide, peptidoglycan, polyriboinosinic-polyribocytidylic acid (Poly I:C), imiquimod, Coley's toxin, polyadenylic-polyuridylic acid (Poly A:U), monophosphoryl lipid A, single-stranded and double-stranded RNA, or CpG oligodeoxynucleotides (ODN).

[0555] Clause C17. The method of any of clauses C1-C16, wherein the administration of a), b), and / or one or more immunomodulatory agents c) is simultaneous, separate or sequential, in any order.

[0556] Clause C18. The method of any of clauses C14-C17, wherein said immunomodulatory agent is administered simultaneously with a therapeutic support composition.

[0557] Clause C19. The method of clause C17 or 18, wherein the simultaneous administration is by co-injection, co-implantation, or co-formulation.

[0558] Clause C20. The method of any of clauses C2-C3 or C5-C19, wherein said immune response is an increase or decrease in one or more of innate and adaptive immune cells.

[0559] Clause C21. The method of any of clauses C2-C3 or C5-C19, wherein said immune response is an increase or decrease in one or more of white blood cells, lymphocytes, monocytes, eosinophils, and antibodies.

[0560] Clause C22. The method of any of clauses C2-C3 or C5-C19, wherein said immune response is an increase in CD3, CD4, CD8, and / or PD-1 positive tumor infiltrating lymphocytes in the first tumor and / or the second tumor.

[0561] Clause C23. The method of any of clauses C2-C3 or C5-C19, wherein said immune response is a reduction in regulatory T cells in the first tumor and / or the second tumor.

[0562] Clause C24. The method of any of clauses C1-C23, wherein the support is a polysaccharide hydrogel, alginate, agarose, cellulose, hyaluronic acid, chitosan, chitin, chondroitin sulfate, heparan sulfate, heparin, gelatin, collagen, a polymer matrix, metal, ceramic, or plastic, each of which may be optionally modified. This specification includes the disclosure of the following inventions. [1] A compound of formula (I) or a pharma- ceutically acceptable salt thereof [ka] (In the formula, R 1a Each time it occurs, C 1~4 Alkyl, hydrogen, and C 1~4 independently selected from the group consisting of haloalkyl; R 1b Each time it occurs, C(O)N(R 1c )-C 1~6 Alkylene-COH, C(O)OH, C(O)N(R 1c )CHR 1e CO2H, C(O)N(R 1c )-C 1~6 Alkylene-C(O)OC 1~4 Alkyl, C(O)OC 1~4 Alkyl, C(O)N(R 1c )CHR 1e C(O)OC 1~4 Alkyl, hydrogen, C 1~4 Alkyl, and C 1~4 independently selected from the group consisting of haloalkyl; R 1c Each occurrence is independently hydrogen or C 1~4 is alkyl; R 1e Each occurrence of is independently -C 1~4 Alkylene-COH, -C1~4 Alkylene-CONH2 or -C 1~4 alkylene-OH; D, at each occurrence, is independently a cyclic dinucleotide; L 1 is, independently at each occurrence, a linker; m is independently at each occurrence 1, 2, or 3; Each occurrence of p is independently 0, 1, or 2. [2] A compound according to formula (IA) [1] or a pharma- ceutically acceptable salt thereof. [ka] . [3] R 1a is hydrogen, or a pharma- ceutically acceptable salt thereof. [4] R 1a is C 1~4 The compound according to [1] or [2], or a pharma- ceutically acceptable salt thereof, wherein R is an alkyl group. [5] R 1a is CH3, or a pharma- ceutically acceptable salt thereof. [6] R 1b is hydrogen, or a pharma- ceutically acceptable salt thereof. [7] R 1b is C(O)N(R 1c )-C 1~6 The compound according to any one of [1] to [5], which is alkylene-CO2H, or a pharma- ceutically acceptable salt thereof. [8] R 1b is C(O)N(R 1c ) CH2CO2H, or a pharma- ceutically acceptable salt thereof. [9] R 1cis hydrogen, or a pharma- ceutically acceptable salt thereof.

[10] R 1b is C(O)OH, or a pharma- ceutically acceptable sa...

Claims

1. A compound of formula (I) or a pharma- ceutically acceptable salt thereof 【Chemistry 1】 (In the formula, R １ａ Each time it occurs, C １～４ Alkyl, hydrogen, and C １～４ independently selected from the group consisting of haloalkyl; R １ｂ Each time it occurs, C(O)N(R １ｃ ) -C １～６ Alkylene-CO ２ H, C(O)OH, C(O)N(R １ｃ ) CHR １ｅ CO ２ H, C(O)N(R １ｃ ) -C １～６ Alkylene-C(O)OC １～４ Alkyl, C(O)OC １～４ Alkyl, C(O)N(R １ｃ ) CHR １ｅ C(O)OC １～４ Alkyl, hydrogen, C １～４ Alkyl, and C １～４ independently selected from the group consisting of haloalkyl; R １ｃ Each occurrence is independently hydrogen or C １～４ is alkyl; R １ｅ Each occurrence independently represents -C １～４ Alkylene-CO ２ H, -C １～４ Alkylene -CONH ２ , or -C １～４ alkylene-OH; D, at each occurrence, is independently a cyclic dinucleotide; L １ is independently at each occurrence a linker; m is independently at each occurrence 1, 2, or 3; Each occurrence of p is independently 0, 1, or 2.

2. The compound according to claim 1 of formula (IA): or a pharma- ceutically acceptable salt thereof. 【Chemistry 2】 。

3. R １ａ is hydrogen; or a pharma- ceutically acceptable salt thereof.

4. R １ａ is C １～４ 3. The compound of claim 1 or 2, or a pharma- ceutically acceptable salt thereof, wherein: R is an alkyl group;

5. R １ａ is CH ３ 3. The compound of claim 1 or 2, wherein:

6. R １ｂ The compound according to any one of claims 1 to 5, or a pharma- ceutically acceptable salt thereof, wherein: is hydrogen.

7. R １ｂ is C(O)N(R １ｃ ) -C １～６ Alkylene-CO ２ H, or a pharma- ceutically acceptable salt thereof.

8. R １ｂ is C(O)N(R １ｃ ) CH ２ CO ２ H, or a pharma- ceutically acceptable salt thereof.

9. R １ｃ The compound according to any one of claims 1 to 5 or 7 to 8, or a pharma- ceutically acceptable salt thereof, wherein:

10. R １ｂ is C(O)OH, or a pharma- ceutically acceptable salt thereof.

11. D is independently 【Chemistry 3】 wherein Y is a nucleobase and X is O or S.

11. The compound according to any one of claims 1 to 10, wherein:

12. 【Chemical 4】 teeth, 【Chemistry 5】 and 12. The compound of any one of claims 1 to 11, or a pharma- ceutically acceptable salt thereof, wherein D' is a cyclic dinucleotide payload moiety.

13. the cyclic dinucleotide payload moiety comprises 【Chemistry 6】 13. The compound of claim 12, wherein:

14. the cyclic dinucleotide payload moiety comprises 【Chemistry 7】 13. The compound of claim 12, wherein:

15. below: 【Chemistry 8】 【Chemistry 9】 【Chemistry 10】 2. The compound of claim 1, selected from the group consisting of:

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

17. A pharmaceutical combination comprising a compound according to any one of claims 1 to 15, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16, for use in the treatment of cancer or for use in enhancing or inducing an immune response, and a therapeutic support composition, said therapeutic support composition comprising a biocompatible support and a compound of formula 【Chemistry 11】 (In the formula, R ２０ is hydrogen, halogen, cyano, nitro, alkyl, alkenyl, alkynyl, heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, cycloalkenyl, CF ３ , C.F. ２ -R', NO ２ , OR', SR', C(=O)R', C(=S)R', OC(=O)R''', SC(=O)R''', OC(=S)R''', SC(=S)R''', S(=O)R', S(=O) ２ R''', S(=O) ２ is selected from the group consisting of NR'R'', C(=O)O-R', C(=O)S-R', C(=S)O-R', C(=S)S-R', C(=O)NR'R'', C(=S)NR'R'', NR'R'', NR'C(=O)R'', NR'C(=S)R'', NR'C(=O)OR'', NR'C(=S)OR'', NR'C(=O)SR'', NR'C(=S)SR'', OC(=O)NR'R'', SC(=O)NR'R'', OC(=S)R'R''', SC(=S)R'R'', NR'C(=O)NR''R'', and NR'C(=S)NR''R''; R' and R'' at each occurrence are independently selected from hydrogen, aryl, and alkyl; R''' at each occurrence is independently selected from aryl and alkyl; R ３０ is halogen, cyano, nitro, hydroxy, alkyl, haloalkyl; alkenyl, alkynyl, alkoxy; haloalkoxy; heteroalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, or cycloalkenyl; R ａ , R ３１ａ and R ３１ｂ are each independently hydrogen, C １ ~C ６ Alkyl, or C １ ~C ６ haloalkyl; t is 0, 1, 2, 3, or 4. A pharmaceutical combination comprising a tetrazine-containing group of formula:

18. 18. The pharmaceutical combination of claim 17, wherein the tetrazine-containing group is linked or directly bonded to a hyaluronic acid biocompatible support.

19. The therapeutic support composition comprises a compound represented by formula (II): 【Chemistry 12】 (In the formula, G ２ teeth, 【Chemistry 13】 and R ２２ is a linker of 1 to 100 linking atoms. The pharmaceutical combination of claim 17, comprising a substituted hyaluronic acid unit of

20. G ２ teeth, 【Chemistry 14】 20. The pharmaceutical combination of claim 19, wherein

21. G ２ teeth, 【Chemistry 15】 and R ２０ is hydrogen or C １～４ 20. The pharmaceutical combination of claim 19, wherein said alkyl is aryl.

22. The pharmaceutical combination according to any one of claims 17 to 21, wherein said use is for the treatment or prevention of cancer.

23. 23. The pharmaceutical combination of claim 22, wherein the cancer is melanoma, renal cancer, prostate cancer, ovarian cancer, breast cancer, glioma, lung cancer, soft tissue cancer, soft tissue sarcoma, osteosarcoma, or pancreatic cancer.

24. 24. The pharmaceutical combination of claim 22 or 23, wherein the cancer is a solid tumor.

25. 24. The pharmaceutical combination of claim 22 or 23, wherein the cancer is a soft tissue sarcoma.

26. 26. The pharmaceutical combination of claim 25, wherein the soft tissue sarcoma is fibrosarcoma, rhabdomyosarcoma, or Ewing's sarcoma.

27. The pharmaceutical combination according to any one of claims 17 to 21, wherein said use is for enhancing or inducing an immune response.

28. 28. The pharmaceutical combination of claim 27, wherein the immune response is an increase in one or more of leukocytes, lymphocytes, monocytes, and eosinophils.

29. The following: an anticancer drug, an immune checkpoint inhibitor, or a compound of formula (IB) or a pharma- ceutically acceptable salt thereof: 【Chemistry 16】 (In the formula, D １ Each occurrence is independently classified as an anticancer drug payload, a microbial immunosuppressant payload, a restenosis inhibitor payload, an antibiotic payload, an antifungal payload, an antiviral payload, a payload selected from an anti-inflammatory / anti-arthritic payload, a corticosteroid payload, and an immunosuppressant payload; R １ａ , R １ｂ , L １ , p and m are as defined in any one of claims 1 to 11. The pharmaceutical combination of any one of claims 17 to 28, further comprising another additional therapeutic agent selected from the group consisting of:

30. p is 0; m is 1; -L １ -teeth, 【Chemistry 17】 30. The pharmaceutical combination of claim 29, wherein

31. 31. The pharmaceutical combination of claim 29 or 30, wherein the anticancer drug is doxorubicin.

32. A kit comprising a compound according to any one of claims 1 to 15, or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition according to claim 16, and instructions for use thereof.

33. The kit of claim 32, further comprising the therapeutic support composition of any one of claims 19 to 23.

34. The kit according to claim 32 or 33, further comprising a compound of formula (IB) according to any one of claims 29 to 31.

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