Targeted steroid compound

Abstract

To provide a compound, or a related compound, comprising folate as a ligand linked to a steroid via a linker.SOLUTION: A compound of formula (I), or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof; a composition comprising such a compound; and the use of such a compound and composition for treating, for example, inflammation associated with a disease or disorder. Formula (I): G1-L-G2, where G1 is a folate radical, an antifolate radical, or a folate analog radical; L is a linker; and G2 is a radical of a steroid.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 958,102, filed January 7, 2020, and U.S. Provisional Patent Application No. 63 / 030,020, filed May 26, 2020, which are incorporated by reference herein in their entireties. [Background technology]

[0002] background Macrophages are a diverse group of white blood cells known to eliminate pathogens through phagocytosis. Macrophages were previously classified according to the organ in which they were found: Kupffer cells in the liver, Langerhans cells in the skin, microglia in the brain and spinal cord, and osteoclasts in bone.

[0003] Current classification of macrophages has shifted from organ-specific macrophages to M1 and M2 macrophages, which is based on macrophage polarization rather than macrophage location.

[0004] M1 macrophages are currently understood to be activated, typically by IFN-γ or lipopolysaccharide (LPS), to produce proinflammatory cytokines, phagocytose microorganisms, and initiate immune responses. M1 macrophages produce nitric oxide (NO) or reactive oxygen intermediates (ROIs) to protect against bacteria and viruses.

[0005] M2 macrophages, in turn, are activated by exposure to certain cytokines, such as IL-4, IL-10, or IL-13. M2 macrophages will produce either polyamines, which induce proliferation, or proline, which induces collagen production. These macrophages are associated with wound healing and tissue repair. Summary of the Invention

[0006] overview The disclosure relates to the design, synthesis, and testing of a series of compounds. Accordingly, the disclosure provides compounds or related compounds comprising folate as a ligand linked to a steroid via a linker. The linker can be any suitable linker, such as a hydrophilic linker. The linker can include one or more amino acids, polyethylene glycol (PEG) monomers, PEG oligomers, PEG polymers, or any combination of the foregoing. The linker can include peptidoglycans, glycans, anions, or oligomers of any combination of the foregoing. The compound polarizes macrophages from a pro-inflammatory (M1) to an anti-inflammatory (M2) state. In alternative embodiments, the ligand is a folate analog or antifolate. The steroid can be any suitable steroid, such as dexamethasone, betamethasone, or betamethasone 17-valerate. The steroid can be releasable, such as via reduction, oxidation, or hydrolysis, or non-releasable. The steroid can be released via a self-immolative moiety.

[0007] Provided in some embodiments herein are compounds of formula (I), or pharmaceutically acceptable salts, polymorphs, prodrugs, solvates, or clathrates thereof, TIFF2025118775000001.tif11128 expression G 1 is a folate radical, antifolate radical, or folate analog radical; L is a linker; and G 2 is a steroid radical.

[0008] Disclosed in some embodiments herein are compounds of formula (I), or pharmaceutically acceptable salts, polymorphs, prodrugs, solvates, or clathrates thereof, TIFF2025118775000002.tif11128 expression G 1 is a folate radical, antifolate radical, or folate analog radical; L is a linker; and G 2 is a radical of a steroid. In some embodiments, the folate radical has the formula: TIFF2025118775000003.tif24128, where the asterisk indicates the point of attachment of the carbonyl carbon to the linker L. In some embodiments, G 1 is a pteroyl-amino acid radical, where the amino acid is selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, arginine, and unnatural amino acids having a derivatizable moiety in the side chain. 1 is an antifolate radical or a folate analog radical comprising an amino acid selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, and arginine. 1 is the radical of any one of the antifolates of the formulae in Table 4 described herein. In some embodiments, the steroid polarizes macrophages from pro-inflammatory (M1) to anti-inflammatory (M2). In some embodiments, G 2 is the radical of a steroid selected from the group consisting of betamethasone, cortisone, cortivazol, difluprednate, hydrocortisone, prednisolone, methylprednisolone, prednisone, dexamethasone, hydrocortisone 17-valerate, budesonide, flumethasone, fluticasone propionate, fluorocortisone, fludrocortisone, paramethasone, eplerenone, and esters of any of the foregoing. 2 is the radical of dexamethasone. In some embodiments, G 2 is the radical of prednisone. In some embodiments, G 2 is a radical of prednisolone. In some embodiments, G 2 is the radical of methylprednisolone. In some embodiments, G 2 is a radical of budesonide. In some embodiments, G 2 is a radical of triamcinolone. In some embodiments, G2 is a radical of betamethasone. In some embodiments, the linker is releasable. In some embodiments, the linker is non-releasable. In some embodiments, the linker comprises one or more of an amino acid, an alkyl chain, a polyethylene glycol (PEG) monomer, a PEG oligomer, a PEG polymer, or a combination of any of the foregoing. In some embodiments, the linker increases the water solubility of the compound. In some embodiments, the linker comprises a peptidoglycan, a glycan, an anion, or an oligomer of a combination of any of the foregoing. In some embodiments, the linker comprises at least one 2,3-diaminopropionic acid group, at least one glutamic acid group, and at least one cysteine group. In some embodiments, the linker has the formula: TIFF2025118775000004.tif26128, where q is an integer from 1 to 10. In some embodiments, the linker comprises a repeating unit of the formula: TIFF2025118775000005.tif28128, where q is an integer from 1 to 10. In some embodiments, the linker has the formula: TIFF2025118775000006.tif36128, wherein X can be O, NH, NR, or S, and q is an integer from 1 to 10, where R is C 1~6 In some embodiments, the linker is of the formula: Includes TIFF2025118775000007.tif47128.

[0009] In some embodiments, the linker is a bivalent linker. In some embodiments, the linker is multivalent, having multiple attachment points for one or more additional chemical groups. In some embodiments, the additional chemical groups are one or more additional G 1 In some embodiments, the additional chemical group comprises a G 1In some embodiments, the linker comprises one or more binding ligands that are not a group. In some embodiments, the linker comprises a PEG oligomer having 2-16 PEG units. In some embodiments, the linker comprises a PEG oligomer having 12 PEG units. In some embodiments, the linker comprises an albumin ligand. In some embodiments, the albumin ligand is Includes TIFF2025118775000008.tif16128.

[0010] In some embodiments, the linker comprises a dimethylcysteine group. In some embodiments, the dimethylcysteine group is linked to a succinimide. Form TIFF2025118775000009.tif29128.

[0011] In some embodiments, the linker comprises a phosphate or pyrophosphate group. In some embodiments, the linker comprises a cathepsin B-cleavable group. In some embodiments, the cathepsin B-cleavable group is valine-citrulline. In some embodiments, the linker comprises a carbamate moiety. In some embodiments, the linker comprises a β-glucuronide. In some embodiments, the linker comprises an ester, phosphate, oxime, acetal, pyrophosphate, polyphosphate, disulfide, sulfate, hydrazide, imine, carbonate, carbamate, or an enzyme-cleavable amino acid sequence. In some embodiments, the linker comprises a self-immolative moiety. In some embodiments, the linker comprises a self-immolative disulfide and / or a sterically protected disulfide bond. In some embodiments, the linker comprises a self-immolative cathepsin-cleavable amino acid sequence. In some embodiments, the linker comprises a self-immolative furin-cleavable amino acid sequence. In some embodiments, the linker comprises a self-immolative β-glucuronidase-cleavable moiety. In some embodiments, the linker comprises a self-immolative phosphatase-cleavable moiety. In some embodiments, the linker comprises a self-immolative sulfatase-cleavable moiety. In some embodiments, the compound has the formula: TIFF2025118775000010.tif21096 or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000011.tif74144, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000012.tif67141, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000013.tif71138 or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000014.tif68141, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000015.tif68128 or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000016.tif88141, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof. In some embodiments, the compound has the formula: TIFF2025118775000017.tif20889 or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. In some embodiments, the compound is selected from the compounds herein.

[0012] In certain embodiments, disclosed herein are pharmaceutical compositions comprising: (a) a compound disclosed herein; and (b) a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutically acceptable excipient is part of a nanoparticle, liposome, or exosome formulation.

[0013] In certain aspects, disclosed herein are methods for transitioning macrophages from M1 to M2 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0014] In certain embodiments, disclosed herein are methods for treating an inflammatory disease or disorder ("disease" and "disorder" may be used interchangeably) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. 2 is the radical of dexamethasone. In some embodiments, G 2 is the radical of prednisone. In some embodiments, G 2 is a radical of prednisolone. In some embodiments, G 2 is the radical of methylprednisolone. In some embodiments, G 2 is a radical of budesonide. In some embodiments, G 2 is a radical of triamcinolone. In some embodiments, G 2 is the radical of betamethasone. In some embodiments, the inflammatory disorder is Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjögren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft-versus-host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor. In some embodiments, the disorder is treated with fewer adverse effects than when the steroid is administered without linkage to a folate ligand.

[0015] In certain embodiments, disclosed herein are methods of treating an autoimmune disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. 2 is the radical of dexamethasone. In some embodiments, G 2 is the radical of prednisone. In some embodiments, G 2 is a radical of prednisolone. In some embodiments, G 2 is the radical of methylprednisolone. In some embodiments, G 2 is a radical of budesonide. In some embodiments, G 2 is a radical of triamcinolone. In some embodiments, G 2 is the radical of betamethasone.

[0016] In certain embodiments, disclosed herein are methods for treating inflammation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. In some embodiments, the inflammation is treated with substantially fewer side effects than when the steroid is administered without linkage to a folate ligand. In some embodiments, the inflammation is associated with an autoimmune disease. In some embodiments, G 2 is the radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, fludrocortisone, beclomethasone, fluticasone, mometasone, ciclesonide, cortisone, cortivazol, hydrocortisone, 21-hydroxypregnenolone, meprednisone, dexamethasone, triamcinolone, betamethasone, prednisone, prednisolone, and methylprednisolone. 2 is the radical of dexamethasone. In some embodiments, G 2 is the radical of prednisone. In some embodiments, G 2is a radical of prednisolone. In some embodiments, G 2 is the radical of methylprednisolone. In some embodiments, G 2 is a radical of budesonide. In some embodiments, G 2 is a radical of triamcinolone. In some embodiments, G 2 is a radical of betamethasone. In some embodiments, the inflammation is associated with Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft-versus-host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor. In some embodiments, the inflammation is associated with Crohn's disease, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, budesonide, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with lupus, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with inflammatory bowel disease, wherein G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, hydrocortisone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with Addison's disease, and G 2is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, hydrocortisone, fludrocortisone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with Graves' disease, G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with Sjogren's syndrome and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with celiac disease, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with Hashimoto's thyroiditis, wherein G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with myasthenia gravis and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with autoimmune vasculitis, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with reactive arthritis, and G 2 is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with psoriatic arthritis, and G 2is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with pernicious anemia, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with ulcerative colitis, and G 2 is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with rheumatoid arthritis and G 2 is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with type 1 diabetes, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with multiple sclerosis, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with asthma, and G 2 is a radical of a steroid selected from the group consisting of triamcinolone, fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with osteoporosis, and G 2is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with sarcoidosis, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with glomerulonephritis, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with autoimmune myocarditis, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with a fibrotic disease, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, and prednisone. In some embodiments, the inflammation is associated with graft-versus-host disease (GVHD), and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with fatty liver disease, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with ischemia-reperfusion injury, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with periprosthetic osteolysis, and G 2is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with scleroderma, and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, budesonide, betamethasone, triamcinolone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with psoriasis, and G 2 is a radical of a steroid selected from the group consisting of budesonide, betamethasone, triamcinolone, prednisone, prednisolone, methylprednisolone, hydrocortisone, dexamethasone, hydrocortisone 17-valerate, diflorasone, meprednisone, halobetacol, tixocortol, amcinonide, desonide, fluocinolone acetonide, fluocinonide, halcinonide, beclomethasone, and halometasone. In some embodiments, the inflammation is associated with spinal cord injury and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is of the central nervous system and G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with a viral infection, and G 2 is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with influenza, and G 2 is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with SARS-CoV-2 (COVID-19), and G 2is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, and prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with cytokine storm syndrome, and G 2 is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with damage to bone, and G 2 Cortisone, Cortivaso In some embodiments, the inflammation is associated with an inflammatory brain disease, and the radical of the steroid is selected from the group consisting of steroids selected from the group consisting of steroids such as steroids, ... 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with atherosclerosis, and G 2 is a radical of a steroid selected from the group consisting of fluticasone, budesonide, beclomethasone, ciclesonide, dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with a tumor, wherein G 2 is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the inflammation is associated with cancer, wherein G 2 is the radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone.

[0017] In certain aspects, disclosed herein are methods for treating a disease or disorder involving polarization of macrophages from M1 to M2 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. [The present invention 1001] A compound of formula (I) or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof: TIFF2025118775000018.tif12128 expression G 1 is a folate radical, antifolate radical, or folate analog radical; L is a linker; and G 2 is a steroid radical. [The present invention 1002] The folate radical has the formula: 1001 compounds of the present invention having the formula TIFF2025118775000019.tif26128, wherein the asterisk indicates the point of attachment of the carbonyl carbon to the linker L. [The present invention 1003] G 1 is a pteroyl-amino acid radical, wherein the amino acid is selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, arginine, and unnatural amino acids having a derivatizable moiety in the side chain. [The present invention 1004] G 1 1001. The compound of claim 1001, wherein is an antifolate radical or a folate analog radical comprising an amino acid selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, and arginine. [The present invention 1005] The compound of the present invention 1001, wherein the steroid polarizes macrophages from pro-inflammatory (M1) to anti-inflammatory (M2). [The present invention 1006] G 21001. A compound of the present invention, wherein R is a radical of a steroid selected from the group consisting of betamethasone, cortisone, cortivazol, difluprednate, hydrocortisone, prednisolone, methylprednisolone, prednisone, dexamethasone, hydrocortisone 17-valerate, budesonide, flumethasone, fluticasone propionate, fluorocortisone, fludrocortisone, paramethasone, eplerenone, and esters of any of the foregoing. [The present invention 1007] G 2 The compound of the present invention 1001, wherein is a radical of dexamethasone. [The present invention 1008] G 2 1001. A compound of the present invention, wherein is a radical of prednisone. [The present invention 1009] G 2 1001. A compound of the present invention, wherein is a radical of prednisolone. [The present invention 1010] G 2 1001. A compound of the present invention, wherein is a radical of methylprednisolone. [The present invention 1011] G 2 1001. A compound of the present invention, wherein is a radical of budesonide. [The present invention 1012] G 2 The compound of the present invention 1001, wherein is a radical of triamcinolone. [The present invention 1013] G 2 The compound of the present invention 1001, wherein is a radical of betamethasone. [The present invention 1014] 1001. A compound of the present invention, wherein the linker is releasable. [The present invention 1015] 1001. A compound of the present invention, wherein the linker is non-releasable. [The present invention 1016] The compound of the present invention 1001, wherein the linker comprises one or more of an amino acid, an alkyl chain, a polyethylene glycol (PEG) monomer, a PEG oligomer, a PEG polymer, or a combination of any of the foregoing. [The present invention 1017] 1001. A compound of the present invention, wherein the linker increases the water solubility of said compound. [The present invention 1018] 1001. The compound of claim 10, wherein the linker comprises an oligomer of a peptidoglycan, a glycan, an anion, or a combination of any of the foregoing. [The present invention 1019] 1001. The compound of claim 10, wherein the linker comprises at least one 2,3-diaminopropionic acid group, at least one glutamic acid group, and at least one cysteine group. [The present invention 1020] The linker has the formula: 1001. A compound of the present invention, comprising a repeating unit of the formula TIFF2025118775000020.tif28128, wherein q is an integer of 1 to 10. [The present invention 1021] The linker has the formula: 1001. A compound of the present invention, comprising TIFF2025118775000021.tif30128, wherein q is an integer of 1 to 10. [The present invention 1022] The linker has the formula: 1001 Compounds of the present invention, including TIFF2025118775000022.tif38128, wherein X can be O, NH, NR, or S, and q is an integer from 1 to 10. [The present invention 1023] The linker has the formula: Compounds of the present invention 1001, including TIFF2025118775000023.tif50128. [The present invention 1024] 1001. The compound of claim 10, wherein the linker is a bivalent linker. [The present invention 1025] The compound of the present invention 1001, wherein the linker is multivalent and has multiple attachment points for one or more additional chemical groups. [The present invention 1026] Additional chemical groups may include one or more additional G 1 The compound of the present invention 1023, comprising a group. [The present invention 1027] Additional chemical groups are G 1 1023. A compound of the present invention comprising one or more binding ligands that are not groups. [The present invention 1028] 1001. The compound of the present invention, wherein the linker comprises a PEG oligomer having 2 to 16 PEG units. [The present invention 1029] 1001. The compound of the present invention, wherein the linker comprises a PEG oligomer having 12 PEG units. [The present invention 1030] 1001. The compound of claim 10, wherein the linker comprises an albumin ligand. [The present invention 1031] Albumin ligand Compounds of the present invention 1028, including TIFF2025118775000024.tif17128. [The present invention 1032] 1001. The compound of claim 10, wherein the linker comprises a dimethylcysteine group. [The present invention 1033] The dimethylcysteine group is linked to succinimide TIFF2025118775000025.tif30128, forming the compound of the present invention 1030. [The present invention 1034] 1001. The compound of claim 10, wherein the linker comprises a phosphate or pyrophosphate group. [This invention 1035] 1001. The compound of claim 10, wherein the linker comprises a cathepsin B cleavable group. [The present invention 1036] 1001. A compound of the present invention, wherein the cathepsin B cleavable group is valine-citrulline. [This invention 1037] 1001. The compound of claim 10, wherein the linker comprises a carbamate moiety. [The present invention 1038] 1001. The compound of claim 10, wherein the linker comprises a β-glucuronide. [This invention 1039] 1001. The compound of claim 10, wherein the linker comprises an ester, phosphate, oxime, acetal, pyrophosphate, polyphosphate, disulfide, sulfate, hydrazide, imine, carbonate, carbamate, or an enzyme-cleavable amino acid sequence. [The present invention 1040] 1001. The compound of claim 10, wherein the linker comprises a self-immolative moiety. [This invention 1041] 1001. The compound of claim 1001, wherein the linker comprises a self-immolative disulfide and / or a sterically protected disulfide bond. [The present invention 1042] 1001. The compound of claim 10, wherein the linker comprises a self-immolative cathepsin-cleavable amino acid sequence. [This invention 1043] 1001. The compound of claim 10, wherein the linker comprises a self-immolative furin-cleavable amino acid sequence. [This invention 1044] 1001. The compound of claim 10, wherein the linker comprises a self-immolative β-glucuronidase cleavable moiety. [This invention 1045] 1001. The compound of claim 1001, wherein the linker comprises a self-immolative phosphatase-cleavable moiety. [The present invention 1046] 1001. The compound of claim 10, wherein the linker comprises a self-immolative sulfatase-cleavable moiety. [This invention 1047] formula: The compound of the present invention 1001 having the formula TIFF2025118775000026.tif227102, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1048] formula: The compound of the present invention 1001 having the formula TIFF2025118775000027.tif79159, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1049] formula: The compound of the present invention 1001 having the formula TIFF2025118775000028.tif71159, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [The present invention 1050] formula: 1001 compounds of the present invention having the formula TIFF2025118775000029.tif76159, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1051] formula: The compound of the present invention 1001 having the formula TIFF2025118775000030.tif72159, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1052] formula: The compound of the present invention 1001 having the formula TIFF2025118775000031.tif72134, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1053] formula: The compound of the present invention 1001 having the formula TIFF2025118775000032.tif80170, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1054] formula: The compound of the present invention 1001 having the formula TIFF2025118775000033.tif22194, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof. [This invention 1055] 1001 compounds of the present invention selected from the compounds described herein. [The present invention 1056] a. any one of compounds 1001 to 1055 of the present invention, b. pharmaceutically acceptable excipients; 10. A pharmaceutical composition comprising: [This invention 1057] 1056. The pharmaceutical composition of the present invention, wherein the pharmaceutically acceptable excipient is part of a nanoparticle, liposomal or exosomal formulation. [This invention 1058] 1. A method for transitioning macrophages from M1 to M2 in a subject in need thereof, comprising: administering a therapeutically effective amount of any of the compounds of the present invention 1001 to 1055 or the composition of the present invention 1056 or 1057 to the subject. A method comprising: [This invention 1059] 1. A method for treating an inflammatory disorder in a subject in need thereof, comprising: administering a therapeutically effective amount of any of the compounds of the present invention 1001 to 1055 or the pharmaceutical composition of the present invention 1056 or 1057 to the subject. A method comprising: [The present invention 1060] G 2 The method of claim 1059, wherein is a radical of dexamethasone. [The present invention 1061] G 2 The method of claim 1059, wherein is a radical of prednisone. [The present invention 1062] G 2 The method of claim 1059, wherein is a radical of prednisolone. [This invention 1063] G 2 The method of claim 1059, wherein is a radical of methylprednisolone. [This invention 1064] G 2 The method of claim 1059, wherein is a radical of budesonide. [This invention 1065] G 2 The method of claim 1059, wherein is a radical of triamcinolone. [The present invention 1066] G2 The method of claim 1059, wherein is a radical of betamethasone. [This invention 1067] 1059. The method of claim 1059, wherein the inflammatory disorder is Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft versus host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor. [The present invention 1068] 1. A method of treating an autoimmune disease in a subject in need thereof, comprising: administering a therapeutically effective amount of any of the compounds of the present invention 1001 to 1055 or the pharmaceutical composition of the present invention 1056 or 1057 to the subject. A method comprising: [The present invention 1069] G 2 The method of claim 1069, wherein is a radical of dexamethasone. [The present invention 1070] G 2 1069. The method of claim 1069, wherein is a radical of prednisone. [This invention 1071] G 2 The method of claim 1069, wherein is a radical of prednisolone. [This invention 1072] G 2 The method of claim 1069, wherein is a radical of methylprednisolone. [This invention 1073] G 2 The method of claim 1069, wherein is a radical of budesonide. [This invention 1074] G 2The method of claim 1069, wherein is a radical of triamcinolone. [This invention 1075] G 2 The method of claim 1069, wherein is a radical of betamethasone. [This invention 1076] 1. A method for treating inflammation in a subject in need thereof, comprising: administering a therapeutically effective amount of any of the compounds of the present invention 1001 to 1055 or the pharmaceutical composition of the present invention 1056 or 1057 to the subject. A method comprising: [This invention 1077] The method of claim 1076, wherein the inflammation is associated with an autoimmune disease. [This invention 1078] G 2 is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, fludrocortisone, beclomethasone, fluticasone, mometasone, ciclesonide, cortisone, cortivazol, hydrocortisone, 21-hydroxypregnenolone, meprednisone, dexamethasone, triamcinolone, betamethasone, prednisone, prednisolone, and methylprednisolone. [This invention 1079] G 2 The method of claim 1076, wherein is a radical of dexamethasone. [The present invention 1080] G 2 The method of claim 1076, wherein is a radical of prednisone. [This invention 1081] G 2 The method of claim 1076, wherein is a radical of prednisolone. [This invention 1082] G 2 The method of claim 1076, wherein is a radical of methylprednisolone. [This invention 1083] G 2 The method of claim 1076, wherein is a radical of budesonide. [This invention 1084] G2 The method of claim 1076, wherein is a radical of triamcinolone. [This invention 1085] G 2 The method of claim 1076, wherein is a radical of betamethasone. [This invention 1086] The method of claim 1076, wherein the inflammation is associated with an immune disorder or disease. [This invention 1087] The method of claim 1076, wherein the inflammation is associated with a neuromuscular disorder or disease. [This invention 1088] The method of claim 1076, wherein the inflammation is associated with a hormonal disorder or disease. [This invention 1089] The method of claim 1076, wherein the inflammation is associated with a gastrointestinal disorder or disease. [The present invention 1090] The method of claim 1076, wherein the inflammation is associated with a connective tissue disease or disorder. [This invention 1091] The method of claim 1076, wherein the inflammation is associated with a liver disease or disorder. [This invention 1092] The method of claim 1076, wherein the inflammation is associated with a musculoskeletal disease or disorder. [This invention 1093] The method of claim 1076, wherein the inflammation is associated with a blood disease or disorder. [This invention 1094] The method of claim 1076, wherein the inflammation is associated with a metabolic disease or disorder. [This invention 1095] The method of claim 1076, wherein the inflammation is associated with a metabolic disease or disorder. [This invention 1096] The method of claim 1076, wherein the inflammation is associated with an endocrine disease or disorder. [This invention 1097] The method of claim 1076, wherein the inflammation is associated with an infection. [This invention 1098] The method of claim 1076, wherein the inflammation is associated with a neurological disease or disorder. [This invention 1099] The method of claim 1076, wherein the inflammation is associated with a kidney disease or disorder. [The present invention 1100] The method of claim 1076, wherein the inflammation is associated with a lung disease or disorder. [The present invention 1101] The method of claim 1076, wherein the inflammation is associated with a tissue disease or disorder. [The present invention 1102] 1076. The method of claim 1076, wherein the inflammation is associated with Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft versus host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor. [The present invention 1103] Inflammation is associated with Crohn's disease, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, budesonide, betamethasone, prednisone, prednisolone, and methylprednisolone. [The present invention 1104] Inflammation is associated with lupus, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1105] Inflammation is associated with inflammatory bowel disease, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, hydrocortisone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1106] Inflammation is associated with Addison's disease, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, hydrocortisone, fludrocortisone, prednisone, prednisolone, and methylprednisolone. [This invention 1107] Inflammation is associated with Graves' disease, 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1108] Inflammation is associated with Sjögren's syndrome, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1109] Inflammation is associated with celiac disease, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1110] Inflammation is associated with Hashimoto's thyroiditis, and G 2 The method of claim 1077, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1111] Inflammation is associated with myasthenia gravis and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1112] Inflammation is associated with autoimmune vasculitis, and G 2The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1113] Inflammation is associated with reactive arthritis, G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1114] Inflammation is associated with psoriatic arthritis, G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1115] Inflammation is associated with pernicious anemia, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [The present invention 1116] Inflammation is associated with ulcerative colitis, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1117] Inflammation is associated with rheumatoid arthritis, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1118] Inflammation is associated with type 1 diabetes, and G 2The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [This invention 1119] Inflammation is associated with multiple sclerosis, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [The present invention 1120] Inflammation is associated with asthma, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of triamcinolone, fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1121] Inflammation is associated with osteoporosis, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [This invention 1122] Inflammation is associated with sarcoidosis, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [This invention 1123] Inflammation is associated with glomerulonephritis, G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1124] Inflammation is associated with autoimmune myocarditis, and G 2The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [Invention 1125] Inflammation is associated with fibrotic disease, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, and prednisone. [The present invention 1126] Inflammation is associated with graft-versus-host disease (GVHD), and 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1127] Inflammation is associated with fatty liver disease, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1128] Inflammation is associated with ischemia-reperfusion injury, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1129] Inflammation is associated with periprosthetic osteolysis, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [The present invention 1130] Inflammation is associated with scleroderma, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, budesonide, betamethasone, triamcinolone, prednisone, prednisolone, and methylprednisolone. [This invention 1131] Inflammation is associated with psoriasis, and G 2 is a radical of a steroid selected from the group consisting of budesonide, betamethasone, triamcinolone, prednisone, prednisolone, methylprednisolone, hydrocortisone, dexamethasone, hydrocortisone 17-valerate, diflorasone, meprednisone, halobetacol, tixocortol, amcinonide, desonide, fluocinolone acetonide, fluocinonide, halcinonide, beclomethasone, and halometasone. [This invention 1132] Inflammation is associated with spinal cord injury, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1133] Inflammation is of the central nervous system, G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1134] Inflammation is associated with viral infection, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1135] Inflammation is associated with influenza, 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1136] Inflammation is associated with SARS-CoV-2 (COVID-19), and G 21076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, and prednisone, prednisolone, and methylprednisolone. [This invention 1137] Inflammation is associated with cytokine storm syndrome, 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1138] Inflammation is associated with damage to bone, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, cortivazol, hydrocortisone, 21-hydroxypregnenolone, meprednisone, dexamethasone, triamcinolone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1139] Inflammation is associated with inflammatory brain diseases, and G 2 The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone. [This invention 1140] Inflammation is associated with atherosclerosis, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of fluticasone, budesonide, beclomethasone, ciclesonide, dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone. [This invention 1141] Inflammation is associated with tumors, where G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone. [This invention 1142] Inflammation is associated with cancer, and G 2 1076. The method of claim 1076, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone. [This invention 1143] and polarizing macrophages from M1 to M2 in a subject in need thereof, comprising: administering a therapeutically effective amount of any of the compounds of the present invention 1001 to 1055 or the pharmaceutical composition of the present invention 1056 or 1057 to the subject. A method comprising: [Brief explanation of the drawings]

[0018] [Figure 1] FIG. 1 illustrates the chemical structure for compound 101 of the disclosure. [Figure 2] FIG. 2 illustrates a reaction scheme for activation of dexamethasone for linking to a ligand comprising folic acid and a moiety that becomes a linker upon reaction with activated dexamethasone. [Figure 3] FIG. 3 illustrates the chemical structure of the ligand shown in FIG. 2, which contains folic acid and a moiety that becomes a linker upon reaction with activated dexamethasone. [Figure 4] FIG. 4 illustrates a reaction scheme for the reaction of activated dexamethasone with the compound of FIG. 3 to form compound 101. [Figure 5] FIG. 5 illustrates the structures and names of the steroids described in the disclosure. [Figure 6] FIG. 6 is a plot of CD206 (a marker for M2 macrophages) expression versus percent of maximum (Max) in flow cytometry for Compound 101 in the presence and absence of the competitor folate-glucosamine. [Figure 7]Figure 7 is a plot of CD206 (a marker for M2 macrophages) expression versus percent of maximum (Max) and shows flow cytometry results for Compound 101 in the presence and absence of the competitor folate-glucosamine compared to untreated cells and cells treated with free dexamethasone. [Figure 8] FIG. 8 is a plot of CD86 (a marker for M1 macrophages) expression versus percent of maximum (Max) and shows flow cytometry results for Compound 101 in the presence and absence of the competitor glucosamine folate, compared to untreated cells and cells treated with free dexamethasone. [Figure 9A] FIG. 9 is a series of plots of flow cytometry results for F4 / 80 goat colon cells in a peritonitis model, including results for Compound 106, untreated, and vehicle-treated cells. [Figure 9B] See legend to Figure 9A. [Figure 9C] See legend to Figure 9A. [Figure 10] FIG. 10 is a bar graph of flow cytometry results for F4 / 80 goat colon cells in a peritonitis model, including results for Compound 106, untreated, and vehicle-treated cells. [Figure 11A] FIG. 11 is a series of plots of flow cytometry results for F4 / 80, CD4, Ly6G, and CD8 goat colon cells in a peritonitis model, including results for Compound 107-treated and untreated cells. [Figure 11B] See legend to Figure 11A. [Figure 11C] See legend to Figure 11A. [Figure 11D] See legend to Figure 11A. [Figure 12]FIG. 12 is a bar graph of flow cytometry results for F4 / 80, CD4, Ly6G, and CD8 goat colon cells in a peritonitis model, including results for Compound 107-treated and untreated cells. [Figure 13A] FIG. 13 is a series of plots of flow cytometry results for F4 / 80, CD4, Ly6G, and CD8 goat colon cells in a peritonitis model, including results for Compound 108-treated and untreated cells. [Figure 13B] See legend to Figure 13A. [Figure 13C] See legend to Figure 13A. [Figure 13D] See legend to Figure 13A. [Figure 14] FIG. 14 is a bar graph of flow cytometry results for F4 / 80, CD4, Ly6G, and CD8 goat colon cells in a peritonitis model, including results for Compound 108-treated and untreated cells. [Figure 15] FIG. 15 is a set of two images of mouse colons, one from an untreated mouse and one from a mouse treated with Compound 107. [Figure 16] FIG. 16 is a series of LC-MS traces for compound 101. [Figure 17A] FIG. 17 is a series of LC-MS traces for compound 106. [Figure 17B] See legend to Figure 17A. [Figure 18A] FIG. 18 is a series of LC-MS traces for compound 107. [Figure 18B] See legend to Figure 18A. [Figure 19A] FIG. 19 is a series of LC-MS traces for compound 125. [Figure 19B] See legend to Figure 19A. [Figure 20A] FIG. 20 is a series of LC-MS traces for compound 108. [Figure 20B]See legend to Figure 20A. [Figure 21A] FIG. 21 is a series of LC-MS traces for compound 124. [Figure 21B] See legend to Figure 21A. [Figure 22A] FIG. 22 is a series of LC-MS traces for compound 126. [Figure 22B] See legend to Figure 22A. [Figure 23A] FIG. 23 is a series of LC-MS traces for compound 127. [Figure 23B] See legend to Figure 23A.

[0019] It should be understood that the drawings are not intended to limit the scope of the present teachings in any way. DETAILED DESCRIPTION OF THE INVENTION

[0020] Detailed Description Folate receptor beta is expressed on the surface of activated macrophages present at sites of inflammation, and these macrophages have been shown to be important in maintaining levels of pro-inflammatory signals in a variety of diseases.

[0021] Steroids work by reducing inflammation and decreasing immune system activity. Inflammation is a process by which the body's white blood cells and chemicals can protect against infection and foreign substances such as bacteria and viruses. In certain diseases, however, the body's defense system (the immune system) does not function properly. This can cause inflammation to work against the body's tissues and cause damage. Signs of inflammation include redness, warmth, swelling, and pain.

[0022] Steroids reduce the production of chemicals that cause inflammation, which helps keep tissue damage as low as possible. Steroids also reduce the activity of the immune system by affecting the way white blood cells work.

[0023] The side effects associated with steroid use vary depending on the dose, type of steroid, and duration of treatment. The following guidelines can help avoid these side effects: use steroids only when necessary, and when possible, use topical steroids for localized problems; use the minimum dose needed to control the disease; and gradually reduce the dose as long as the disease is controlled.

[0024] The above side effects can be minimized by targeted delivery of steroids to the site of inflammation. For example, steroids can be delivered to activated macrophages by compounds that target the cell surface folate receptor beta. To date, the inventors have not seen any evidence in the literature of the synthesis and use of folic acid as a steroid-linked ligand. Therefore, there is an unmet need for such compounds.

[0025] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art of chemistry and biology. Additionally, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural of the referent unless the context clearly dictates otherwise. Thus, for example, if a compound / composition is substituted with "an" alkyl or aryl, the compound / composition may also be substituted with at least one alkyl and / or at least one aryl. Furthermore, unless otherwise specified, the term "about" refers to a range of plus or minus 10% for percentages and plus or minus 1.0 units for unit values; for example, about 1.0 refers to a range of 0.9 to 1.1.

[0026] When a chemical group is combined with several other chemical groups as defined herein, each part of the combination is considered to be defined as such when separated, taking into account valences to allow for attachment of other groups. For example, an "alkoxycycloalkylenecarbonyl" radical is understood to be an alkoxy, as defined herein, attached to a cycloalkylene, as defined herein, which in turn is attached to a carbonyl group, not defined herein but commonly understood by organic chemists, with an open valence on the carbonyl.

[0027] The term "radical" as used herein refers to a molecular fragment, where the fragment has an open valence for bond formation. A monovalent radical has one open valence so that it can form a bond with another chemical group. In some embodiments, a molecular radical (e.g., a steroid radical) as used herein is generated by removing one hydrogen atom from the molecule to generate a monovalent radical with one open valence at the location of the removed hydrogen atom. Where appropriate, the radical can be divalent, trivalent, etc., where two, three, or more hydrogen atoms are removed to generate a radical that can bond to two, three, or more chemical groups. Where appropriate, the open valence of the radical can be generated by removing a non-hydrogen atom (e.g., a halogen atom) or by removing two or more atoms (e.g., a hydroxyl group), as long as the removed atoms are only a small fraction (20% or less of the total number of atoms) of the total atoms in the radical-forming molecule. In some embodiments, the radical is formed from a folic acid, antifolate, or folic acid analog by removal of a hydroxyl group.

[0028] As used herein, the term "substituted" or "substituent" refers to a group that can be or is substituted on a molecule or other group (e.g., an aryl or alkyl group). Examples of substituents include halogen (e.g., F, Cl, Br, and I), OR, OC(O)N(R), CN, NO, NO, ONO, azido, CF, OCF, R, O(oxo), S(thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R), SR, SOR, SO, R, SO, N(R), SO, R, -(CH). 0~2 P(O)(OR)2, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2) 0~2 N(R)C(O)R, (CH2) 0~2 N(R)C(O)OR, (CH2) 0~2 Examples include, but are not limited to, N(R)N(R), N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R), N(R)SOR, N(R)SON(R), N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R), N(R)C(S)N(R), N(COR)COR, N(OR)R, C(=NH)N(R), C(O)N(OR)R, or C(=NOR)R, where each R is independently hydrogen, alkyl, It can be acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, where any alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, or two R groups attached to a nitrogen atom or adjacent nitrogen atoms, together with the nitrogen atom or nitrogen atoms, can form a heterocyclyl, which can be mono- or independently polysubstituted.

[0029] As used herein, an "individual," "subject," or "patient" is a mammal, preferably a human, but can also be an animal.

[0030] "Oxo" refers to the =O radical.

[0031] "Alkyl" refers to a straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms, such as those having 1 to 15 carbon atoms (e.g., C1-C 15 The disclosure provided herein of "alkyl" is intended to include the independent description of saturated "alkyl" unless otherwise specified. In certain embodiments, alkyl contains 1 to 13 carbon atoms (e.g., C1-C 13 In certain embodiments, an alkyl comprises 1 to 8 carbon atoms (e.g., a C1-C8 alkyl). In other embodiments, an alkyl comprises 1 to 5 carbon atoms (e.g., a C1-C5 alkyl). In other embodiments, an alkyl comprises 1 to 4 carbon atoms (e.g., a C1-C4 alkyl). In other embodiments, an alkyl comprises 1 to 3 carbon atoms (e.g., a C1-C3 alkyl). In other embodiments, an alkyl comprises 1 to 2 carbon atoms (e.g., a C1-C2 alkyl). In other embodiments, an alkyl comprises 1 carbon atom (e.g., a C1 alkyl). In other embodiments, an alkyl comprises 5 to 15 carbon atoms (e.g., a C5-C 15 In another embodiment, an alkyl group contains 5 to 8 carbon atoms (e.g., a C5-C8 alkyl). In another embodiment, an alkyl group contains 2 to 5 carbon atoms (e.g., a C2-C5 alkyl). In another embodiment, an alkyl group contains 3 to 5 carbon atoms (e.g., a C3-C5 alkyl). In another embodiment, an alkyl group is selected from the group consisting of methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (isobutyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). An alkyl is attached to the remainder of the molecule by a single bond.

[0032] "Alkoxy" refers to a radical attached through an oxygen atom of the formula --O-alkyl, where alkyl is an alkyl chain as defined above.

[0033] "Alkylene" or "alkylene chain" generally refers to a straight- or branched-chain divalent alkyl group, such as those having 1 to 12 carbon atoms, that attaches the remainder of the molecule to a radical group, e.g., methylene, ethylene, propylene, i-propylene, n-butylene, etc.

[0034] "Aryl" refers to a radical derived from an aromatic monocyclic or polycyclic hydrocarbon ring system by removing a hydrogen atom from a ring carbon atom. The aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and carbon atoms from 5 to 18 carbon atoms, where at least one of the rings within the ring system is fully unsaturated, i.e., contains a cyclic delocalized (4n+2) π-electron system according to the Hückel theory. Ring systems from which aryl groups are derived include, but are not limited to, groups such as benzene, fluorene, indane, indene, tetralin, and naphthalene.

[0035] An "aralkyl" or "aryl-alkyl" is a group of the formula -R c -aryl radical, where R c is an alkylene chain as defined above, for example, methylene, ethylene, etc. The alkylene chain part of the aralkyl radical may be optionally substituted as described above for an alkylene chain.

[0036] "Cycloalkyl" refers to a stable 3- to 18-membered non-aromatic cyclic radical containing only carbon atoms as ring atoms. Unless otherwise specified in the specification, a cycloalkyl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, optionally including aromatic, fused, and / or bridged ring systems. Examples of such radicals include cyclopropyl, cyclohexyl, norbornyl, and adamantyl. As used herein, "cycloalkylene" specifically refers to a divalent cycloalkyl radical.

[0037] "Halo" or "halogen" refers to a bromo, chloro, fluoro, or iodo substituent.

[0038] "Haloalkyl" refers to an alkyl radical, as defined above, that is substituted by one or more halogen radicals, as defined above, e.g., trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like.

[0039] The term "heteroalkyl" refers to an alkyl group, as defined above, in which one or more skeletal carbon atoms of the alkyl have been replaced with a heteroatom (with the appropriate number of substituents or valences, e.g., -CH- may be replaced with -NH- or -O-). For example, each substituted carbon atom is independently replaced with a heteroatom, e.g., carbon replaced with nitrogen, oxygen, selenium, or other suitable heteroatom. In some cases, each substituted carbon atom is independently replaced with oxygen, nitrogen (e.g., with -NH-, -N(alkyl)-, or -N(aryl)- or other substituents contemplated herein), or sulfur (e.g., -S-, -S(=O)-, or -S(=O)-). In some embodiments, a heteroalkyl is attached to the remainder of the molecule at a carbon atom of the heteroalkyl. In some embodiments, a heteroalkyl is attached to the remainder of the molecule at a heteroatom of the heteroalkyl. In some embodiments, a heteroalkyl is a C1-C 18 In some embodiments, heteroalkyl is C1-C 12 Heteroalkyl is. In some embodiments, heteroalkyl is C1-C6 heteroalkyl. In some embodiments, heteroalkyl is C1-C4 heteroalkyl. Heteroalkyl can include alkoxy, alkoxyalkyl, alkylamino, alkylaminoalkyl, aminoalkyl, heterocycloalkyl, and heterocycloalkylalkyl, as defined herein.

[0040] "Heteroalkylene" refers to a divalent heteroalkyl group, as defined above, linking one moiety of the molecule to another moiety of the molecule.

[0041] "Heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring radical containing 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified in the specification, a heterocyclyl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include aromatic, fused, and / or bridged ring systems. The heteroatoms in a heterocyclyl radical may be oxidized. The heterocyclyl radical may be partially or fully saturated. The disclosure provided herein for "heterocyclyl" is intended to include independent descriptions of heterocyclyl, including aromatic and non-aromatic ring structures, unless otherwise specified. A heterocyclyl is attached to the remainder of the molecule through any atom of the ring. Examples of such heterocyclyl radicals are dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, 1,3-benzodioxolyl, 1,4-benzodioxanyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, indolinyl, isoindolinyl In some embodiments, the aryl group may be substituted or unsubstituted, including, but not limited to, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0042] "Heteroaryl" refers to a radical derived from a 3- to 18-membered aromatic ring radical containing 2 to 17 carbon atoms and 1 to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. As used herein, a heteroaryl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, wherein at least one of the rings within the ring system is fully unsaturated, i.e., contains a cyclic delocalized (4n+2) π-electron system according to Hückel theory. Heteroaryl includes fused or bridged ring systems. Heteroatoms in a heteroaryl radical may be oxidized. If present, one or more nitrogen atoms may be quaternized. A heteroaryl is attached to the remainder of the molecule through any atom of the ring. Examples of heteroaryl include azepinyl, acridinyl, benzimidazolyl, benzoindolyl, benzofuranyl, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, and 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl. , 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7 ,8,9,10-Hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10-Hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolyl, indolizinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl, 1,6-Naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyri Examples of thiazolyl include, but are not limited to, pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e., thienyl).

[0043] As used herein, the term "heterocycloalkyl" refers to an alkyl group, as defined herein, in which a hydrogen or carbon bond of the alkyl group, as defined herein, is replaced with a bond to a heterocyclyl group, as defined herein. Representative heterocycloalkyl groups include, but are not limited to, furan-2-ylmethyl, furan-3-ylmethyl, pyridin-3-ylmethyl, tetrahydrofuran-2-ylmethyl, and indol-2-ylpropyl. As used herein, the term "heterocycloalkylalkyl" refers to a heterocycloalkyl group bonded to an alkyl group, as defined herein.

[0044] The term "heteroarylalkyl," as used herein, refers to an alkyl group, as defined herein, in which a hydrogen or carbon bond of the alkyl group is replaced with a bond to a heteroaryl group, as defined herein.

[0045] As used herein, the term "amine" refers to, for example, a group of formula N(group)3 +where each group may independently be H or non-H, e.g., alkyl, aryl, etc. Amines include, but are not limited to, R-NH, e.g., alkylamines, arylamines, alkylarylamines; RNH, where R is defined herein, e.g., dialkylamines, diarylamines, aralkylamines, heterocyclylamines, etc.; and RN, where each R is independently selected, e.g., trialkylamines, dialkylarylamines, alkyldiarylamines, triarylamines, etc. The term "amine" as used herein also includes ammonium ions.

[0046] As used herein, the term "amino group" refers to any of -NH2, -NHR, -NR2, and -NR3, where each R is as defined herein. + Substituents of the form -NR3, and -NR3 cannot be protonated + " refers to the respective protonated forms except for the following: ##STR1## Thus, any compound substituted with an amino group may be considered an amine. An "amino group" within the meaning herein may be a primary, secondary, tertiary, or quaternary amino group. An "alkylamino" group includes monoalkylamino, dialkylamino, and trialkylamino groups.

[0047] Examples of "alkylamino" are -NH-alkyl and -N(alkyl)2.

[0048] The compounds disclosed herein, in some embodiments, contain one or more asymmetric centers, thus giving rise to enantiomers, diastereomers, and other stereoisomers defined in terms of absolute stereochemistry as (R)- or (S)-. Unless otherwise specified, all stereoisomeric forms of the compounds disclosed herein are intended to be contemplated by the present disclosure. When a compound described herein contains an alkene double bond, unless otherwise specified, the present disclosure is intended to include both E and Z geometric isomers (e.g., cis or trans). Similarly, all possible isomers, including their racemic and optically pure forms, and all tautomeric forms, are also intended to be included. The term "geometric isomer" refers to the E or Z geometric isomer (e.g., cis or trans) of the alkene double bond. The term "positional isomer" refers to structural isomers around a central ring, such as ortho, meta, and para isomers around a benzene ring.

[0049] As used herein, the term "linker" generally refers to a G 1 (e.g., binding ligand) and / or G 2 A "linker" refers to a portion of a compound that forms a chemical bond with a folate ligand (e.g., a therapeutic agent). In particular, a "linker" can connect two or more functional portions of a molecule to form a compound provided herein. Illustratively, a linker can include atoms selected from C, N, O, S, Si, and P; C, N, O, S, and P; or C, N, O, and S. A linker can connect different functional portions of a compound, such as a folate ligand and a therapeutic agent. A linker can include several linker groups in a contiguous backbone, for example, ranging from about 2 to about 100 atoms. In some embodiments, a linker is a releasable linker. In some embodiments, a linker is a non-releasable linker.

[0050] As used herein, a "pteroyl" radical, moiety, or group has the following structure: TIFF2025118775000034.tif24128, where the asterisk represents the point of attachment of the carbonyl carbon to another chemical group, such as a linker, L.

[0051] As used herein, a "pteroyl-amino acid" radical, moiety, or group has the following structure: TIFF2025118775000035.tif24128, where the asterisk represents the point of attachment of the carbonyl carbon to another chemical group, such as a linker L, and H2N-Ax-COOH is an amino acid.

[0052] As used herein, "reducing adverse events" in a subject refers to a reduction in the incidence and / or severity of adverse events occurring in treated patients, where the reduction is significant at at least a 95% confidence level. As used herein, "adverse events" include serious events that cause death, hospitalization, and / or disability (temporary or permanent), as well as less serious events that cause discomfort perceptible by the subject (skin rash, sleep disturbance, hair loss, headache, etc.).

[0053] As used herein, the terms "salt" and "pharmaceutically acceptable salt" refer to derivatives of the disclosed compounds in which the parent compound is modified by making its acid or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic groups such as amines; alkali or organic salts of acidic groups such as carboxylic acids. Pharmaceutically acceptable salts include the conventional non-toxic salts or quaternary ammonium salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid; and salts prepared from organic acids such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, and isethionic acid.

[0054] Pharmaceutically acceptable salts can be synthesized from parent compounds that contain basic or acidic moieties by conventional chemical methods.In some cases, such salts can be prepared by reacting the free acid or base form of these compounds with stoichiometric amounts of suitable base or acid in water or organic solvent, or in the mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.A list of suitable salts can be found in, for example, Chapter 38 of Remington's Pharmaceutical Sciences, 21st ed., Lippincott Williams & Wilkins, 2006, the disclosure of which is incorporated herein by reference.

[0055] The term "solvate" means a compound or a salt thereof that further includes a stoichiometric or non-stoichiometric amount of solvent bound by non-covalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0056] The term "prodrug" refers to a derivative of a compound that can be hydrolyzed, oxidized, or reacted in some way under biological conditions (in vitro or in vivo) to provide an active compound, particularly the compounds disclosed herein. Examples of prodrugs include, but are not limited to, derivatives and metabolites of the compounds disclosed herein that contain biohydrolyzable moieties such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureides, and biohydrolyzable phosphate analogs. Specific prodrugs of compounds with a carboxyl functional group are lower alkyl esters of the carboxylic acid. Carboxylic acid esters are conveniently formed by esterifying any of the carboxylic acid moieties present on the molecule. Prodrugs can typically be prepared using well-known methods, such as those described by Burger's Medicinal Chemistry and Drug Discovery 6th ed. (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers GmbH).

[0057] The term "polymorph" generally refers to crystalline substances that have the same chemical composition but different molecular packing. The term "crystalline salt" includes crystalline structures that have the same chemical composition but incorporate an acid or base addition salt within the molecular packing of the crystalline structure.

[0058] The term "clathrate" generally refers to a compound in which molecules of one component (eg, a solvent) are physically trapped within the crystal structure of another component.

[0059] The terms "non-releasable linker" and "non-cleavable linker" are used interchangeably. As used herein, they refer to linkers that cannot be cleaved under extracellular physiological conditions (e.g., pH-labile, acid-labile, oxidation-labile, or enzyme-labile bonds). However, such linkers may contain bonds that can be cleaved after entering a cell.

[0060] As used herein, the term "releasable linker" refers to a linker that contains at least one bond (e.g., a pH-labile, acid-labile, oxidation-labile, or enzyme-labile bond) that can be cleaved under extracellular physiological conditions. Releasable groups also include photochemically cleavable groups. Examples of photochemically cleavable groups include linkers containing 2-(2-nitrophenyl)-ethan-2-ol groups, and o-nitrobenzyl, decyl, trans-o-cinnamoyl, m-nitrophenyl, or benzylsulfonyl groups (see, for example, Dorman and Prestwich, Trends Biotech. 18:64-77 (2000); and Greene and Wuts, Protective Groups in Organic Synthesis, 2nd ed., John Wiley & Sons, New York (1991)).

[0061] The cleavable bond may be present within the cleavable linker and / or at one or both termini of the cleavable linker. It should be appreciated that physiological conditions that result in bond cleavage include, for example, standard chemical hydrolysis reactions that occur at physiological pH or as a result of compartmentalization into organelles, such as endosomes, that have a pH lower than the cytoplasmic pH. Illustratively, the bivalent linkers described herein can undergo cleavage under other physiological or metabolic conditions, such as through the action of glutathione-mediated mechanisms. It should be appreciated that the lability of the cleavable bond can be adjusted by including functional groups or fragments within the bivalent linker L that can assist or promote such bond cleavage, also referred to as neighboring group assistance. The lability of the cleavable bond can also be adjusted by, for example, substitutional modifications at or near the cleavable bond, such as including an alpha branch adjacent to the cleavable disulfide bond, increasing the hydrophobicity of the silicon-on-oxygen bond-containing moiety, homologating an alkoxy group to form a hydrolyzable ketal or acetal moiety, etc. In addition, it is recognized that, if present, additional functional groups or fragments may be included within the bivalent linker L that can assist or facilitate further fragmentation of the compound following bond cleavage of the releasable linker.

[0062] As used herein, the terms "subject," "patient," and "individual" are used interchangeably. Neither term is intended to require the ongoing supervision of a medical professional. The subject may be any mammal, e.g., a human.

[0063] As used herein, the term "treating" includes therapeutic treatment (e.g., a subject having signs and symptoms of the condition being treated) and / or prophylactic treatment. Prophylactic treatment includes preventing as well as inhibiting or slowing the progression of the condition.

[0064] As used herein, the term "therapeutically effective amount" refers to an amount of one or more compounds of the various embodiments described herein (e.g., a compound of Formula (I)) that produces the biological or medical response in a tissue system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician, including alleviation of the signs or symptoms of the disease or disorder being treated.

[0065] As used herein, an "antifolate" is a compound that binds to the folate receptor and antagonizes the biological action of folic acid or one of its naturally occurring forms, such as dihydrofolate, 5-methyltetrahydrofolate, or methylenetetrahydrofolate. Antifolates include inhibitors of dihydrofolate reductase, thymidylate synthase, and other enzymes in the folate biosynthetic pathway. Antifolates include methotrexate, pemetrexed, proguanil, pyrimethamine, ralitrexate, pralatrexate, and trimethoprim. Antifolates further include the compounds set forth in Table 4 herein.

[0066] As used herein, the terms "a," "an," or "the" are used to include one or more unless the context clearly dictates otherwise. The term "or" is used to refer to a non-exclusive "or" unless otherwise indicated. Additionally, any phrases or terms employed herein and not specifically defined should be understood to be for purposes of description only, not limitation. Any use of section headings is intended to aid in the reading and comprehension of the document and should not be construed as limiting. Additionally, information associated with a section heading may appear within or outside of that particular section.

[0067] As used herein, the term "about" allows for some variation in a value or range, for example, within 10%, within 5%, or within 1% of a stated value or stated range.

[0068] The terms and expressions employed are used as terms of description and not of limitation. In this regard, if a particular term is defined under "Definitions" and is otherwise defined, explained, or discussed anywhere in the "Detailed Description," all such definitions, explanations, and discussions are intended to be ascribed to such terms. Nor is the use of such terms and expressions intended to exclude all equivalents to the features shown and described or portions thereof. Furthermore, while subheadings such as "Definitions" are used in the "Detailed Description," such use is for ease of reference only and is not intended to limit any disclosure provided in one section to only that section; rather, any disclosure provided under one subheading is intended to apply to the disclosures under all other subheadings as well.

[0069] compound In one aspect, the present disclosure relates to the design, synthesis, and testing of a series of compounds. Thus, there is provided a compound of formula (I), or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof: TIFF2025118775000036.tif11128 expression G 1 is a folate radical, antifolate radical, or folate analog radical; L is a linker; and G 2 is a steroid radical.

[0070] In some embodiments, the folate radical has the formula: TIFF2025118775000037.tif25128, where the asterisk (*) indicates the point of attachment of the carbonyl carbon to the linker L. In an alternative embodiment, G 1 may be the radical of an antifolate (e.g., a folate antagonist or folic acid antagonist) or a folate analog radical. 1is the radical of an antifolate of any one of the formulas in Table 4 described herein.

[0071] In some embodiments, G 1 is a folate analog radical. In some embodiments, the folate analog is a pteroyl moiety or pteroyl-amino acid moiety or folate analog of any one of formulas V-X described herein. In some embodiments, G 1 is a pteroyl-amino acid radical. In some embodiments, G 1 is a pteroyl-amino acid radical, where the amino acid is selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, arginine, and unnatural amino acids having a derivatizable moiety in the side chain.

[0072] In some embodiments, G 1 is an antifolate radical or a folate analog radical comprising an amino acid selected from the group consisting of aspartic acid, lysine, tyrosine, cysteine, threonine, serine, histidine, and arginine. 1 is a radical of a folic acid analog of any one of formulas V-X described herein. In some embodiments, G 1 is the radical of an antifolate of any one of the formulas in Table 4 described herein.

[0073] Provided in some embodiments herein are compounds of formula (I), or pharmaceutically acceptable salts, polymorphs, prodrugs, solvates, or clathrates thereof, TIFF2025118775000038.tif11128 expression G 1 is the expression: TIFF2025118775000039.tif24128 folate radical, where the asterisk indicates the point of attachment of the carbonyl carbon to the linker L; or pteroyl-amino acid radical; L is a linker; and G2 is the radical of a corticosteroid.

[0074] In some embodiments, G 1 is a radical of a folate analog, which is a pyrido[2,3-d]pyrimidine analog ligand, functional fragment, or analog thereof, having affinity (e.g., without limitation, high specificity) for the folate receptor. For example, such a folate analog can have a relative affinity for binding a folate receptor, such as folate receptor β (FRβ), of about 0.01 or greater compared to folic acid at about 20°C / 25°C / 30°C / physiological temperature.

[0075] Specific examples of suitable targeting moieties (or radicals thereof) are provided herein; however, the G 1 It is understood that the targeting moiety (or radical thereof) can include any ligand (or radical thereof) useful for targeting the folate receptor and is not limited to the structures identified herein. The ligand (or radical thereof) can bind to the folate receptor.

[0076] As used herein, for example, in Formulas V-XI below, a group depicted with a bond extending into a ring but not terminating at a ring atom (such as C(O)Z) is understood to be a substituent replacing any hydrogen bonded to a ring atom of that ring. This includes any hydrogen that is explicitly shown or described (e.g., X in Formulas V-XI). 1 ~X 9 hydrogen as part of the group), and hydrogen substitutions that are not explicitly shown but are understood to be present.

[0077] In some embodiments, G 1 has the structure of formula V or a radical thereof, TIFF2025118775000040.tif38128 expression X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamic acid or valine; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m and n are each independently 0, 1, or 0 to 1; and TIFF2025118775000041.tif3128 represents either a single or double bond CC.

[0078] In some embodiments, Formula V has the structure of VI, or a radical thereof: TIFF2025118775000042.tif36128 expression X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamic acid or valine; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m and n are each independently 0, 1, or 0 to 1; and TIFF2025118775000043.tif3128 represents either a single or double bond CC.

[0079] In some embodiments, Formula V has the structure of Formula VII, or a radical thereof: TIFF2025118775000044.tif33128 expression X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamic acid or valine; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m and n are each independently 0, 1, or 0 to 1; and TIFF2025118775000045.tif3128 represents either a single or double bond CC.

[0080] In some embodiments, Formula V has the structure of Formula VIII, or a radical thereof: TIFF2025118775000046.tif34128 expression X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamic acid or valine; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m is 0, 1, or 0 to 1; and TIFF2025118775000047.tif3128 represents either a single or double bond CC.

[0081] In some embodiments, Formula V has the structure of Formula IX, or a radical thereof: TIFF2025118775000048.tif41128 expression X1, X2, X3, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamic acid or valine; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m is 0, 1, or 0 to 1; and TIFF2025118775000049.tif3128 represents either a single or double bond CC.

[0082] In some embodiments, Formula V has the structure of Formula X, or a radical thereof: TIFF2025118775000050.tif36128 expression X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamate, valine, or substrate; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is H or alkyl; m is 0, 1, or 0 to 1; and TIFF2025118775000051.tif3128 represents either a single or double bond CC.

[0083] In some embodiments, Formula V has the structure of Formula XI, or a radical thereof: TIFF2025118775000052.tif40128 expression X1, X2, X3, X4, X5, X6, X7, X8, and X9 are each independently N, NH, CH, CH2, O, or S; Y is C, CH, CH2, N, NH, O, or S; Z is glutamate, valine, or substrate; R1 and R2 are each independently NH2, OH, SH, CH3, or H; R3 is alkyl or alkyl; m is 0, 1, or 0 to 1; and TIFF2025118775000053.tif3128 represents either a single or double bond CC.

[0084] G of this disclosure 1Chemical structures and spectroscopic data for some additional embodiments of moieties (or radicals thereof, etc.) are provided in Tables 1, 2, 3, and 4 below.

[0085] Table 1 provides non-limiting examples of additional embodiments of Formula VIII.

[0086] (Table 1) Formula VIII TIFF2025118775000054.tif72128TIFF2025118775000055.tif224126TIFF2025118775000056.tif126128

[0087] Table 2 provides non-limiting examples of additional embodiments of Formula IX.

[0088] (Table 2) Formula IX TIFF2025118775000057.tif188128

[0089] Table 3 provides non-limiting examples of additional embodiments of Formula X.

[0090] (Table 3) Formula X TIFF2025118775000058.tif140128

[0091] In some embodiments, instead of folic acid, G 1 The moiety (e.g., radical thereof) is a non-classical antifolate (or analog or functional fragment thereof) having a formula (e.g., radical of the formula) set forth in Table 4 below, such as, for example, a pyrido[2,3-d]pyrimidine or similar analog (or radical thereof). Note that in Table 4, the 2',3'-(C4H4) substituent indicates an aryl ring fused at the 2' and 3' positions of the substituted aryl ring to form a bicyclic aryl system.

[0092] Table 4. Non-classical antifolate analogs TIFF2025118775000059.tif84128TIFF2025118775000060.tif207126TIFF2025118775000061.tif220126TIFF2025118775000062.tif130128

[0093] The steroid present in the compounds described herein can be any suitable steroid that polarizes macrophages from pro-inflammatory (M1) to anti-inflammatory (M2). 2 is the radical of a corticosteroid. Examples of such corticosteroids include betamethasone, cortisone, cortivazol, difluprednate, hydrocortisone, prednisolone, methylprednisolone, prednisone, dexamethasone, hydrocortisone 17-valerate, fluorocortisone, fludrocortisone, paramethasone, eplerenone, amcinonide, alclometasone, beclomethasone, ciclesonide, clobetasol, clocortolone, desonide, deflazacort, and desoximetazol. flucloxone, diflorasone, fluocinolone, fluocinonide, fluprednidene, flunisolide, fluticasone, fluchlorolone, fludroxycortide, fluorometholone, flucorolone, halcinonide, halometasone, 21-hydroxypregnenolone, halobetacol, loteprednol, meprednisone, mometasone, prednicarbate, prebediolone, rimexolone, tixocortol, triamcinolone, or esters of any of the foregoing.

[0094] In some embodiments, G 2is the radical of a steroid selected from the group consisting of betamethasone, cortisone, cortivazol, difluprednate, hydrocortisone, prednisolone, methylprednisolone, prednisone, dexamethasone, hydrocortisone 17-valerate, budesonide, flumethasone, fluticasone propionate, fluorocortisone, fludrocortisone, paramethasone, eplerenone, and esters of any of the foregoing. In some embodiments, the steroid is betamethasone. In some embodiments, the steroid is cortisone. In some embodiments, the steroid is cortivazol. In some embodiments, the steroid is difluprednate. In some embodiments, the steroid is hydrocortisone. In some embodiments, the steroid is prednisolone. In some embodiments, the steroid is methylprednisolone. In some embodiments, the steroid is prednisone. In some embodiments, the steroid is dexamethasone. In some embodiments, the steroid is hydrocortisone 17-valerate. In some embodiments, the steroid is budesonide. In some embodiments, the steroid is flumethasone. In some embodiments, the steroid is fluticasone propionate. In some embodiments, the steroid is fluorocortisone. In some embodiments, the steroid is fludrocortisone. In some embodiments, the steroid is paramethasone. In some embodiments, the steroid is eplerenone. In some embodiments, the steroid is amcinonide. In some embodiments, the steroid is alclometasone. In some embodiments, the steroid is beclomethasone. In some embodiments, the steroid is ciclesonide. In some embodiments, the steroid is clobetasol. In some embodiments, the steroid is clocortolone. In some embodiments, the steroid is desonide. In some embodiments, the steroid is deflazacort. In some embodiments, the steroid is desoximetasone. In some embodiments, the steroid is diflorasone. In some embodiments, the steroid is fluocinolone. In some embodiments, the steroid is fluocinonide.In some embodiments, the steroid is fluprednidene. In some embodiments, the steroid is flunisolide. In some embodiments, the steroid is fluticasone. In some embodiments, the steroid is fluchlorolone. In some embodiments, the steroid is fludroxycortide. In some embodiments, the steroid is fluorometholone. In some embodiments, the steroid is flucorolone. In some embodiments, the steroid is halcinonide. In some embodiments, the steroid is halometasone. In some embodiments, the steroid is 21-hydroxypregnenolone. In some embodiments, the steroid is halobetacol. In some embodiments, the steroid is loteprednol. In some embodiments, the steroid is meprednisone. In some embodiments, the steroid is mometasone. In some embodiments, the steroid is prednicarbate. In some embodiments, the steroid is prebedilone. In some embodiments, the steroid is rimexolone. In some embodiments, the steroid is tixocortol. In some embodiments, the steroid is triamcinolone.

[0095] The linker present in the compounds described herein can be any suitable linker. For example, the linker can be a hydrophilic linker, such as a linker that can include one or more of an amino acid (which can be the same or different), an alkyl chain, a polyethylene glycol (PEG) monomer, a PEG oligomer, a PEG polymer, or any combination of the foregoing. The linker can include a peptidoglycan, a glycan, or an anionic oligomer. In some embodiments, when the linker includes a chemical group, the group includes one or more of its atoms in the backbone of the linker. In some embodiments, the chemical group does not need to include atoms in the backbone of L if the group is for binding purposes (such as an albumin binding group), a glucuronide, or a "W" group as described herein. For linkers that include one or more PEG units, all carbon and oxygen atoms of the PEG units are part of the backbone unless otherwise specified. For releasable linkers, the cleavable bond is part of the backbone. The "backbone" of the linker L is G. 1 and G 2 In some embodiments, the polyvalent linker has a branched backbone, with each branch acting as a portion of the backbone linker until a terminal end is reached.

[0096] The L groups described herein can have any suitable length and chemical composition. For example, L can have a chain length of at least about 7 atoms long. In some embodiments, L is at least about 10 atoms long. In some embodiments, L is at least about 14 atoms long. In some embodiments, L is about 7 to about 31 atoms long (e.g., about 7 to 31, 7 to about 31, or 7 to 31), about 7 to about 24 atoms long (e.g., about 7 to 24, 7 to about 24, or 7 to 24), or about 7 to about 20 atoms long (e.g., about 7 to 20, 7 to about 20, or 7 to 20). In some embodiments, L is about 14 to about 31 atoms in length (e.g., about 14 to 31, 14 to about 31, or 14 to 31), about 14 to about 24 atoms in length (e.g., about 14 to 24, 14 to about 24, or 14 to 24), or about 14 to about 20 atoms in length (e.g., about 14 to 20, 14 to about 20, or 14 to 20). In some embodiments, L has a chain length of at least 7 atoms, at least 14 atoms, at least 20 atoms, at least 25 atoms, at least 30 atoms, at least 40 atoms; or 1 to 15 atoms, 1 to 5 atoms, 5 to 10 atoms, 5 to 20 atoms, 10 to 40 atoms, or 25 to 100 atoms. Examples of L linker groups having a chain length of 1 to 5 atoms are of the formula: TIFF2025118775000063.tif15128, wherein R z1 is H, alkyl, arylalkyl, or -alkyl-S-alkyl, or the side chain of any naturally occurring or non-naturally occurring amino acid, etc.; the number represents the atom that is counted as part of the chain, in this case 3 atoms. z1 Examples of R include H (i.e., the side chain of glycine), alkyl (e.g., the side chain of alanine, valine, isoleucine, and leucine), -alkyl-S-alkyl (e.g., the side chain of methionine), arylalkyl (e.g., the side chain of phenylalanine, tyrosine, and tryptophan), and the like. z1 The atoms to which is attached may be chiral and may have any suitable relative configuration, such as D- or L-configuration.

[0097] The atoms used in forming L can be combined in any chemically relevant manner, such as a chain of carbon atoms forming an alkylene group, a chain of carbon and oxygen atoms forming a polyoxyalkylene group, a chain of carbon and nitrogen atoms forming a polyamine, etc. In addition, it is understood that the bonds connecting the atoms in the chain can be either saturated or unsaturated, such that, for example, alkanes, alkenes, alkynes, cycloalkanes, arylenes, imides, etc. can be divalent radicals included in L. In addition, the atoms forming the linker can also be of the formula: TIFF2025118775000064.tif15128 (in the formula, each 5 are independently CH2, N(X 5 (if there is a bond connected to X), NH, or O, and each X 6 are independently N, C(X 6 It is understood that groups may be cyclized with each other to form saturated or unsaturated bivalent cyclic radicals in the linker, such as radicals of the formula: Such as that in TIFF2025118775000065.tif36128. In each of the foregoing and other L groups described herein, the chain forming the linker may be substituted or unsubstituted.

[0098] Alternatively, or in addition to chain length, in some embodiments, L has suitable substituents that can affect the hydrophobicity or hydrophilicity of L. Thus, for example, L can have a hydrophobic side chain group such as an alkyl, cycloalkyl, aryl, arylalkyl, etc. group, each of which may be substituted. When L comprises one or more amino acids, L can contain hydrophobic amino acid side chains, such as one or more amino acid side chains from phenylalanine (Phe) and tyrosine (Tyr), including substitution variants, and analogs and derivatives of such side chains. These side chain variants, analogs, and derivatives include, for example, TIFF2025118775000066.tif18128, which are variants of tyrosine, amine analogs of tyrosine, and methoxy derivatives of tyrosine, respectively. Other variants, analogs, and derivatives are contemplated.

[0099] In some embodiments, L comprises a moiety that is neutral under physiological conditions. In some embodiments, L comprises a moiety that can be protonated or deprotonated to bear one or more positive charges or one or more negative charges, respectively. In some embodiments, L comprises a neutral moiety and a moiety that can be protonated to bear one or more positive charges. Examples of neutral moieties include sugars, carbohydrates, saccharides, polyhydroxyl groups such as inositol, and / or polyether groups such as polyoxyalkylene groups including polyoxyethylene, polyoxypropylene, and the like. Examples of moieties that can be protonated to bear one or more positive charges include amino groups such as polyaminoalkylenes including ethylenediamine, propylenediamine, butylenediamine, and the like, and / or heterocycles containing pyrrolidine, piperidine, piperazine, and other amino groups, each of which may be substituted. Examples of moieties that can be deprotonated to bear one or more negative charges include carboxylic acids such as aspartic acid, glutamic acid, and long-chain carboxylic acid groups, and sulfate esters such as alkyl esters of sulfuric acid.

[0100] Exemplary polyoxyalkylene groups include those with specific length ranges of about 4 to about 20 polyoxyalkylene (e.g., polyethylene glycol) groups, e.g., about 4-20, 4 to about 20, or 4-20 polyoxyalkylene groups. Exemplary alkyl sulfates may also be directly incorporated into the backbone via click chemistry. Exemplary L groups containing polyamines include EDTA and DTPA radicals: TIFF2025118775000067.tif31128(Poly)peptide: TIFF2025118775000068.tif15128β-amino acids, etc.: TIFF2025118775000069.tif16128 and combinations thereof, wherein each R 31 are independently H, alkyl, arylalkyl, heterocyclylalkyl, ureido, aminoalkyl, alkylthio, or amidoalkyl, as in the side chains of naturally occurring amino acids such as alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, tryptophan, serine, threonine, asparagine, methionine, lysine, arginine, and histidine. Non-naturally occurring amino acids are also contemplated herein.

[0101] As discussed herein, in some embodiments, L comprises at least one releasable moiety. In some embodiments, L comprises at least two releasable linkers (e.g., cleavable linkers). The choice of releasable or non-releasable linker can be made independently for each use or configuration of the compounds described herein. The releasable linkers described herein include various atoms, chains of atoms, functional groups, and combinations of functional groups. For example, in some embodiments, the releasable linker comprises about 1 to about 30 atoms (e.g., about 1 to about 30 atoms, 1 to about 30 atoms, and 1 to 30 atoms), or about 2 to about 20 atoms (e.g., about 2 to 20 atoms, 2 to about 20 atoms, and 2 to 20 atoms). Low molecular weight linkers (e.g., those having an approximate molecular weight of about 30 g / mol to about 1,000 g / mol, e.g., about 30 g / mol to about 300 g / mol, about 100 g / mol to about 500 g / mol, or about 150 g / mol to about 600 g / mol) are also described. Precursors of such linkers may be selected to have either nucleophilic or electrophilic functional groups or both, optionally in protected form with conveniently cleavable protecting groups to facilitate their use in the synthesis of intermediate species.

[0102] In some embodiments, the linker is a bivalent linker (e.g., a single G 1 A single G 2In some embodiments, the linker is a polyvalent linker (e.g., a linker that connects two or more G 1 A single G 2 In some embodiments, the linker is multivalent and has multiple points of attachment for one or more additional chemical groups (e.g., the additional chemical groups are connected to one or more additional G 1 or the additional chemical group is G 1 (including one or more attached ligands that are not groups). In some embodiments, the linker is a releasable linker. In some embodiments, the linker is a non-releasable linker.

[0103] In some embodiments, L is (L 1 ) o -Y-(L 2 ) p wherein: Each L 1 is the first linker; Each L 2 is a second linker; Y is a template connecting the arms of the compound; o is an integer from 1 to 5; and p is an integer of 1 to 5.

[0104] In some embodiments, L 1 and L 2 are identical. In some embodiments, L 1 and L 2 In some embodiments, each L 1 is G 1 In certain embodiments, each L 2 is G 2 In certain embodiments, o and m are the same, e.g., 1 to 6, 1 to 3, or 1. In some embodiments, p is 1. In some embodiments, o is 1. In some embodiments, p and o are each 1.

[0105] In some embodiments, each L 1 and L 2independently comprise an oligoethylene glycol (chain), a polyethylene glycol (chain), an alkyl (chain), an oligopeptide (chain), or a polypeptide (chain). In some embodiments, each L 1 and L 2 independently comprise an oligoethylene glycol (chain) or a polyethylene glycol (chain).

[0106] In some embodiments, each L 1 and L 2 independently comprises a triazole or an amide.

[0107] In some embodiments, each L 1 and L 2 In some embodiments, each L 1 and L 2 Each independently contains a peptidoglycan (chain).

[0108] In some embodiments, each L 1 and L 2 independently comprise an oligoproline or an oligopiperidine.

[0109] In some embodiments, each L 1 and L 2 are independently 15 to 200 angstroms (Å) in length.

[0110] In some embodiments, o is an integer from 1 to 5. In some embodiments, o is an integer from 1 to 3. In some embodiments, o is 1. In some embodiments, o is m.

[0111] In some embodiments, p is an integer from 1 to 5. In some embodiments, p is an integer from 1 to 3. In some embodiments, p is 1.

[0112] The following formula: Provided in some embodiments herein are polyvalent compounds having the formula: G1 is a folate radical, antifolate radical, or folate analog radical; S is a spacer (e.g., having a length that allows the arms of a multivalent targeting ligand (e.g., a drug) to reach multiple adjacent folate receptors on a target cell); Y is a template connecting the arms of the compound; L is G 1 G 2 (e.g., connect L to G 1 a first covalent bond connecting L to G 2 (e.g., bifunctional) linker (through a second covalent bond linking G 2 is a steroid radical; and m is 2 to 6.

[0113] In some embodiments, the spacer is of an optimal length for the arms of the multivalent drug to reach multiple adjacent folate receptors on the target (eg, macrophage) cell.

[0114] In some embodiments, S comprises an oligoethylene, polyethylene glycol, an alkyl chain, an oligopeptide, or a polypeptide. In some embodiments, S is an oligoethylene glycol or polyethylene glycol.

[0115] In some embodiments, S is an oligopeptide or polypeptide.

[0116] In some embodiments, S is peptidoglycan.

[0117] In some embodiments, the spacer is a rigid linker. In some embodiments, S is a rigid linker such as, for example, an oligoproline or an oligopiperidine.

[0118] In some embodiments, S is at least 15 Angstroms (Å) in length. In some embodiments, S is up to 200 Angstroms (Å) in length. In some embodiments, S is between 15 and 200 Angstroms (Å) in length.

[0119] In some embodiments, Y is a template connecting multiple arms of the compound. In some embodiments, Y has a repeating structure. In some embodiments, Y comprises a releasable bond. In some embodiments, L comprises a disulfide bond. In some embodiments, Y comprises at least one citrate group (or radical thereof). In some embodiments, Y comprises one or more triazoles. In some embodiments, Y comprises one or more amines. In some embodiments, Y comprises one or more amides. In some embodiments, Y has an aromatic core (e.g., an aryl core or a heteroaryl core). In some embodiments, Y has an alkyl(ene) core. In some embodiments, Y has an amine core. In some embodiments, Y is N(L 1 ) 3 (e.g., where L 1 (as described elsewhere herein). In some embodiments, Y is selected from the group consisting of three L 1 (e.g., where L 1 are described elsewhere herein). In some embodiments, Y is C(L 1 ) 4 (e.g., where L 1 are described elsewhere herein).

[0120] In some embodiments, Y is one L 1 In some embodiments, Y binds to one L 2 In some embodiments, Y binds to one L 1 and one L 2 In some embodiments, Y is independently L 1 and L 2 In some embodiments, Y is attached to L, and L is attached to Y via an amide bond.

[0121] In some embodiments, Y is a template (e.g., a multivalent template) that connects multiple arms of the compound. In some embodiments, Y has a repeating structure. In some embodiments, Y includes at least one citrate group (or radical thereof). In some embodiments, the template has the following structure: TIFF2025118775000071.tif29128

[0122] In some embodiments, Y is a template that connects multiple arms of the compound (e.g., a multivalent template) and includes a template (e.g., a repeat unit) of the following structure: TIFF2025118775000072.tif63128

[0123] In some embodiments, Y is a template that connects multiple arms of a compound having a citrate-based template. In some embodiments, Y is a template that connects multiple arms of a compound (e.g., a multivalent template) and has a (e.g., citrate-based) template of the following structure: TIFF2025118775000073.tif50128

[0124] In some embodiments, Y is a template that connects multiple arms of the compound (e.g., a multivalent template), having a (e.g., citrate-based) template of the following structure: TIFF2025118775000074.tif60128

[0125] In some embodiments, Y is a template that connects multiple arms of the compound (e.g., a multivalent template), having a (e.g., citrate-based) template of the following structure: TIFF2025118775000075.tif37128

[0126] In some embodiments, L comprises at least one linker group, each linker group selected from the group consisting of polyethylene glycol (PEG), alkyl, sugar, and peptide. In some embodiments, the linker is a polyethylene glycol (PEG) (e.g., pegylated), alkyl, sugar, and peptide-based dual linker.

[0127] In some embodiments, the linker comprises a PEG oligomer having 2 to 16 PEG units, hi some embodiments, the linker comprises a PEG oligomer having 12 PEG units.

[0128] In some embodiments, L is a non-releasable linker (e.g., G 2 and G 1 In some embodiments, L is a releasable linker (e.g., G 2 and G 1 is bivalently (e.g., covalently) bound to

[0129] In one example, L includes one or more releasable linkers that cleave under the conditions described herein by a chemical mechanism involving beta-elimination. Such releasable linkers include beta-thio, beta-hydroxy, and beta-amino substituted carboxylic acids and their derivatives, such as esters, amides, carbonates, carbamates, and ureas. Such linkers also include 2- and 4-thioaryl esters, carbamates, and carbonates.

[0130] An example of a releasable linker is a linker of the formula: TIFF2025118775000076.tif21128, wherein X 4 is NR, n is an integer selected from 0, 1, 2, and 3, and R 32 is hydrogen or a substituent that contains a substituent capable of stabilizing a positive charge inductively or by resonance on the aryl ring, such as alkoxy. The releasable linker may be further substituted.

[0131] Assisted cleavage of the releasable moiety of L can include mechanisms involving a benzylium intermediate, a benzyne intermediate, lactone cyclization, an oxonium intermediate, beta-elimination, and the like. In addition to fragmentation following cleavage of the releasable moiety of L, the initial cleavage of the releasable linker can be facilitated by an anchimerically assisted mechanism. Thus, in the example of the releasable moiety of L listed above, the hydroxyalkanoic acid, which may be cyclized, facilitates cleavage of the methylene bridge, e.g., by an oxonium ion, and promotes bond cleavage or subsequent fragmentation after bond cleavage of the releasable linker. Alternatively, acid-catalyzed oxonium ion-assisted cleavage of the methylene bridge can initiate a series of fragmentations of this exemplary bivalent linker or its fragments. Alternatively, acid-catalyzed hydrolysis of the carbamate can facilitate beta-elimination of the hydroxyalkanoic acid, which may cyclize and promote cleavage of the methylene bridge, e.g., by an oxonium ion. It is recognized that other chemical mechanisms of bond cleavage under the metabolic, physiological, or cellular conditions described herein can initiate such a series of fragmentations. It is recognized that other chemical mechanisms of bond cleavage under the metabolic, physiological, or cellular conditions described herein may initiate such a series of fragmentations.

[0132] Exemplary mechanisms for cleavage of the bivalent linkers described herein include the following 1,4 and 1,6 fragmentation mechanisms for carbonates and carbamates: TIFF2025118775000077.tif84128 during the ceremony, Nuc - is an exogenous or endogenous nucleophile, glutathione, or a bioreductant, and R a and X a is connected through the other part of the bivalent linker. a and X a The position of, for example, the resulting product is X a -S-Nuc and HO-R a H2N-R a The value can be switched to:

[0133] While the above fragmentation mechanisms are illustrated as concerted mechanisms, any number of individual steps may occur to result in the final fragmentation of the bivalent linker to the depicted end product. For example, bond cleavage can also occur via acid-catalyzed elimination of the carbamate moiety, which may be anchimerically assisted by stabilization provided by either the beta-sulfur or disulfide aryl group shown in the above examples. In these variations of this embodiment, the releasable linker is the carbamate moiety. Alternatively, fragmentation can be initiated by nucleophilic attack on the disulfide group, resulting in the formation of a thiolate upon cleavage. The thiolate can intermolecularly displace a carbonate or carbamate moiety to form the corresponding thiacyclopropane. In the case of a benzyl-containing bivalent linker, following exemplary cleavage of the disulfide bond, the resulting phenyl thiolate can further fragment to liberate the carbonate or carbamate moiety by forming a resonance-stabilized intermediate. In any of these cases, the releasable nature of the exemplary bivalent linkers described herein may be achieved by any mechanism that may be related to the chemical, metabolic, physiological, or biological conditions present.

[0134] Thus, as described above, the releasable linker can comprise a disulfide group. Further examples of releasable linkers within L include divalent radicals comprising an alkyleneaziridin-1-yl, alkylenecarbonylaziridin-1-yl, carbonylalkylaziridin-1-yl, alkylenesulfoxylaziridin-1-yl, sulfoxylalkylaziridin-1-yl, sulfonylalkylaziridin-1-yl, or alkylenesulfonylaziridin-1-yl group, wherein each of the releasable linkers is optionally substituted. Additional examples of releasable linkers contained in L include divalent radicals comprising methylene, 1-alkoxyalkylene, 1-alkoxycycloalkylene, 1-alkoxyalkylenecarbonyl, 1-alkoxycycloalkylenecarbonyl, carbonylarylcarbonyl, carbonyl(carboxyaryl)carbonyl, carbonyl(biscarboxyaryl)carbonyl, haloalkylenecarbonyl, alkylene(dialkylsilyl), alkylene(alkylarylsilyl), alkylene(diarylsilyl), (dialkylsilyl)aryl, (alkylarylsilyl)aryl, (diarylsilyl)aryl, oxycarbonyloxy, oxycarbonyloxyalkyl, sulfonyloxy, oxysulfonylalkyl, iminoalkylidenyl, carbonylalkylideneiminyl, iminocycloalkylidenyl, carbonylcycloalkylideneiminyl, alkylenethio, alkylenearylthio, or carbonylalkylthio groups, wherein each of the releasable linkers is optionally substituted.

[0135] Additional examples of releasable linkers included in L include an oxygen atom and a methylene, 1-alkoxyalkylene, 1-alkoxycycloalkylene, 1-alkoxyalkylenecarbonyl, or 1-alkoxycycloalkylenecarbonyl group, where each of the releasable linkers is optionally substituted. Alternatively, in some embodiments, the releasable linker includes an oxygen atom and a methylene group, where the methylene group is substituted with an optionally substituted aryl, and the releasable linker is bonded to the oxygen to form an acetal or ketal. Furthermore, in some embodiments, the releasable linker includes an oxygen atom and a sulfonylalkyl group, where the releasable linker is bonded to the oxygen to form an alkylsulfonate.

[0136] Additional examples of releasable linkers within L include nitrogen and iminoalkylidenyl, carbonylalkylideneiminyl, iminocycloalkylidenyl, and carbonylcycloalkylideneiminyl groups, each of which is optionally substituted, where the releasable linker is attached to the nitrogen to form a hydrazone. In some embodiments, the hydrazone is acylated with a carboxylic acid derivative, an orthoformic acid derivative, or a carbamoyl derivative to form various acylhydrazone releasable linkers.

[0137] Additional examples of releasable linkers included in L include an oxygen atom and an alkylene(dialkylsilyl), alkylene(alkylarylsilyl), alkylene(diarylsilyl), (dialkylsilyl)aryl, (alkylarylsilyl)aryl, or (diarylsilyl)aryl group, each of which is optionally substituted, and in which the releasable linker is bonded to the oxygen to form a silanol.

[0138] Additional examples of releasable linkers within L include two independent nitrogens and a carbonylarylcarbonyl, a carbonyl(carboxyaryl)carbonyl, or a carbonyl(biscarboxyaryl)carbonyl. In some embodiments, the releasable linker is attached to the heteroatom nitrogen to form an amide and is connected to X through an amide bond. a or R a Combine with.

[0139] Additional examples of releasable linkers included in L include an oxygen atom, a nitrogen atom, and a carbonylarylcarbonyl, a carbonyl(carboxyaryl)carbonyl, or a carbonyl(biscarboxyaryl)carbonyl. In some embodiments, the releasable linker forms an amide, and in some embodiments, is connected to X via an amide bond. a or R a Combine with.

[0140] In some embodiments, L comprises an optionally substituted 1-alkylenesuccinimide-3-yl group and a releasable moiety comprising a methylene, 1-alkoxyalkylene, 1-alkoxycycloalkylene, 1-alkoxyalkylenecarbonyl, or 1-alkoxycycloalkylenecarbonyl group, each of which is optionally substituted, to form a succinimide-1-yl alkyl acetal or ketal.

[0141] In some embodiments, L comprises carbonyl, thionocarbonyl, alkylene, cycloalkylene, alkylenecycloalkyl, alkylenecarbonyl, cycloalkylenecarbonyl, carbonylalkylcarbonyl, 1-alkylenesuccinimide-3-yl, 1-(carbonylalkyl)succinimide-3-yl, alkylenesulfoxyl, sulfonylalkyl, alkylenesulfoxylalkyl, alkylenesulfonylalkyl, carbonyltetrahydro-2H-pyranyl, carbonyltetrahydrofuranyl, 1-(carbonyltetrahydro-2H-pyranyl)succinimide-3-yl, or 1-(carbonyltetrahydrofuranyl)succinimide-3-yl, each of which is optionally substituted. In some embodiments, L further comprises an additional nitrogen such that L comprises an alkylenecarbonyl, cycloalkylenecarbonyl, carbonylalkylcarbonyl, or 1-(carbonylalkyl)succinimide-3-yl group attached to the nitrogen, each of which is optionally substituted, to form an amide. In some embodiments, L further comprises a sulfur atom and an alkylene or cycloalkylene group, each optionally substituted with carboxy, attached to the sulfur to form a thiol, hi some embodiments, L comprises a sulfur atom and a 1-alkylenesuccinimide-3-yl and 1-(carbonylalkyl)succinimide-3-yl group attached to the sulfur to form a succinimide-3-yl thiol.

[0142] In some embodiments, L comprises a nitrogen and a releasable moiety comprising an alkyleneaziridin-1-yl, carbonylalkylaziridin-1-yl, sulfoxylalkylaziridin-1-yl, or sulfonylalkylaziridin-1-yl, each of which is optionally substituted. In some embodiments, L comprises a carbonyl, thionocarbonyl, alkylenecarbonyl, cycloalkylenecarbonyl, carbonylalkylcarbonyl, or 1-(carbonylalkyl)succinimide-3-yl, each of which is optionally substituted, bonded to a releasable moiety to form an aziridine amide.

[0143] Examples of L include the following formula: As further illustrated by TIFF2025118775000078.tif21128, examples include alkylene-amino-alkylenecarbonyl, alkylene-thio-(carbonylalkylsuccinimide-3-yl), and the like, wherein x' and y' are each independently 1, 2, 3, 4, or 5.

[0144] L is C (e.g., -CH2-, C(O)), N (e.g., NH, NR b , where R b can have any suitable combination of atoms in the chain, including, for example, H, alkyl, alkylaryl, etc.), O (e.g., —O—), P (e.g., —OP(O)(OH)O—), and S (e.g., —S—). For example, the atoms used in forming L can be combined in all chemically relevant ways, such as chains of carbon atoms forming alkyl groups, chains of carbon and oxygen atoms forming polyoxyalkyl groups, chains of carbon and nitrogen atoms forming polyamines, and other ring-containing chains such as those forming aryl and heterocyclyl groups (e.g., triazoles, oxazoles, etc.). In addition, the bonds connecting the atoms in the chain in L can be either saturated or unsaturated, such that, for example, alkanes, alkenes, alkynes, cycloalkanes, arylenes, imides, etc. can be divalent radicals included in L. Furthermore, the Ls forming the chain can be substituted or unsubstituted.

[0145] Additional examples of L groups include 1-alkylsuccinimide-3-yl, carbonyl, thionocarbonyl, alkyl, cycloalkyl, alkylcycloalkyl, alkylcarbonyl, cycloalkylcarbonyl, carbonylalkylcarbonyl, 1-alkylsuccinimide-3-yl, 1-(carbonylalkyl)succinimide-3-yl, alkylsulfoxyl, sulfonylalkyl, alkylsulfoxylalkyl, alkylsulfonylalkyl, carbonyltetrahydro-2H-pyranyl, carbonyltetrahydrofuranyl, 1-(carbonyltetrahydro-2H-pyranyl)succinimide-3-yl, and 1-(carbonyltetrahydrofuranyl)succinimide-3-yl, where each group can be substituted or unsubstituted. Any of the above groups can be L or can be included as part of L. In some cases, any of the above groups may be used in combination (or more than once) (e.g., -alkyl-C(O)-alkyl), and may further include an additional nitrogen (e.g., alkyl-C(O)-NH-, -NH-alkyl-C(O)-, or -NH-alkyl-), oxygen (e.g., -alkyl-O-alkyl-), or sulfur (e.g., -alkyl-S-alkyl-). Examples of such L groups are alkylcarbonyl, cycloalkylcarbonyl, carbonylalkylcarbonyl, 1-(carbonylalkyl)succinimide-3-yl, and succinimide-3-ylthiol, where each group may be substituted or unsubstituted.

[0146] In some embodiments, L is formed via / derived from click chemistry. Those skilled in the art will appreciate that the terms "click chemistry" and "click chemistry-derived" generally refer to a class of small molecule reactions commonly used in conjugation, allowing for the attachment of selected substrates to specific molecules. Click chemistry is not a single specific reaction, but rather describes a method for generating products following examples from nature, which also generate substances by combining small modular units. In many applications, click reactions link biomolecules to reporter molecules. Click chemistry is not limited to biological contexts, and the "click" reaction concept has been used in pharmacological and various biomimetic applications. However, it has been found to be particularly useful for the detection, localization, and qualification of biomolecules.

[0147] The Click reaction can occur in one pot, is typically not hindered by water, generates minimal by-products, and is characterized by a high thermodynamic driving force that rapidly and irreversibly drives high yields of a single reaction product with high reaction specificity (in some cases, both regiospecificity and stereospecificity). These properties make the Click reaction well suited to the challenge of isolating and targeting molecules in complex biological environments. In such environments, therefore, the product must be physiologically stable, and any by-products must be non-toxic (as in in vivo systems).

[0148] Examples of click chemistry include copper-catalyzed azide-alkyne cycloaddition (CuAAC), strain-promoted azide-alkyne cycloaddition (SPAAC), inverse electron-demand Diels-Alder reaction (IEDDA), and Staudinger ligation (SL), which can derive L. For example, X can be synthesized as shown in Schemes 1-5. a and R a can be linked together. TIFF2025118775000079.tif166128In formula, each R bis independently H, alkyl, arylalkyl, -alkyl-S-alkyl or arylalkyl, or the side chain of any naturally occurring or non-naturally occurring amino acid, etc. In Schemes 1-5, X a and R a The wavy line connecting to is X a and R a and the groups to which they are attached. In Schemes 1-5, it is understood that triazole, oxazole, and -NH-SO2-NH- groups can be considered part of L.

[0149] In some embodiments, L is a linker selected from the group consisting of pegylated, alkyl, sugar, and peptide-based dual linkers. L is either a non-releasable or releasable linker bivalently covalently attached to a folate ligand (or, in another embodiment, a folate analog or antifolate) and a steroid.

[0150] In some embodiments, L is TIFF2025118775000080.tif32128, where x'' is an integer between 0 and 10, and y'' is an integer from 3 to 100.

[0151] In some aspects, x'' is an integer from 3 to 10.

[0152] In some embodiments, L is TIFF2025118775000081.tif86128, where R 33 and R 34 are each independently H or C1-C6 alkyl; and z is an integer from 1 to 8.

[0153] In some embodiments, L is TIFF2025118775000082.tif18128.

[0154] In some embodiments, L is TIFF2025118775000083.tif35128, where R 37 is H or C1-C6 alkyl; R 35a , R 35b , R 36a , and R 36b are each independently H or C1-C6 alkyl.

[0155] In some embodiments, L comprises an amino acid selected from the group consisting of Lys, Asn, Thr, Ser, Ile, Met, Pro, His, Gln, Arg, Gly, Asp, Glu, Ala, Val, Phe, Leu, Tyr, Cys, and Trp. In some embodiments, L comprises at least two amino acids independently selected from the group consisting of Glu and Cys. In some embodiments, L comprises Glu-Glu, where the glutamic acids are covalently linked to each other through their carboxylic acid side chains.

[0156] In some embodiments, L comprises one or more hydrophilic spacer linkers comprising multiple hydroxyl functional groups.

[0157] In some embodiments, L comprises at least one 2,3-diaminopropionic acid group, at least one glutamic acid group (e.g., the unnatural amino acid D-glutamic acid), and at least one cysteine group. An example of such a linker has the formula: TIFF2025118775000084.tif27128, where q is an integer between 1 and 10 (e.g., between 1 and 3 and between 2 and 5). In some embodiments, L has the general formula: TIFF2025118775000085.tif70128, wherein X can be O, NH, NR, or S, and q is an integer from 1 to 10. In some embodiments, L is a group of the formula: TIFF2025118775000086.tif48128, in which the disulfide group is part of a self-immolative group that can be generically described as a group of the formula -CH2-SS-CH2-.

[0158] In some embodiments, the compounds described herein comprise a linkage that allows the steroids described herein to be released by any suitable mechanism, including release mechanisms involving reduction, oxidation, or hydrolysis. An example of a reduction mechanism involves the reduction of a disulfide group into two separate sulfhydryl groups. Thus, for example, a group of formula -CH2-SS-CH2- can be used when the linker has the formula: TIFF2025118775000087.tif48128, the reduction product has the formula: TIFF2025118775000088.tif49128, which will be reduced to two separate groups of formula -CH2-SH. In this example, the steroid is attached to the linker via a self-immolative moiety (e.g., a disulfide group).

[0159] Examples of self-immolative disulfides also include sterically protected disulfide bonds. The steroid can be linked to the linker via any other suitable self-immolative bond, including via a self-immolative cathepsin-cleavable amino acid sequence; via a self-immolative furin-cleavable amino acid sequence; via a self-immolative β-glucuronidase-cleavable moiety; via a self-immolative phosphatase-cleavable moiety; or via a self-immolative sulfatase-cleavable moiety. Multiple self-immolative linkages are also contemplated herein.

[0160] In some embodiments, the linker comprises a self-immolative moiety. In some embodiments, the linker comprises a self-immolative disulfide and / or a sterically protected disulfide bond. In some embodiments, the linker comprises a self-immolative cathepsin-cleavable amino acid sequence. In some embodiments, the linker comprises a self-immolative furin-cleavable amino acid sequence. In some embodiments, the linker comprises a self-immolative β-glucuronidase-cleavable moiety. In some embodiments, the linker comprises a self-immolative phosphatase-cleavable moiety. In some embodiments, the linker comprises a self-immolative sulfatase-cleavable moiety.

[0161] In some embodiments, the linker comprises a phosphate or pyrophosphate group. In some embodiments, the linker comprises a cathepsin B cleavable group. In some embodiments, the cathepsin B cleavable group is valine-citrulline. In some embodiments, the linker comprises a carbamate moiety. In some embodiments, the linker comprises a β-glucuronide.

[0162] In some embodiments, the compounds described herein include a linkage in which the steroid is attached to the linker via an ester, phosphate, oxime, acetal, pyrophosphate, polyphosphate, disulfide, sulfate, hydrazide, imine, carbonate, carbamate, or an enzyme-cleavable amino acid sequence.

[0163] In some embodiments, the linker comprises an ester, phosphate, oxime, acetal, pyrophosphate, polyphosphate, disulfide, sulfate, hydrazide, imine, carbonate, carbamate, or an enzyme-cleavable amino acid sequence.

[0164] In some embodiments, L comprises one or more spacer linkers. The spacer linker can be a hydrophilic spacer linker containing multiple hydroxyl functional groups. The spacer "L" can comprise any stable atomic arrangement. The spacer comprises one or more L'. Each L' is independently selected from the group consisting of an amide, an ester, a urea, a carbonate, a carbamate, a disulfide, an amino acid, an amine, an ether, an alkyl, an alkene, an alkyne, a heteroalkyl (e.g., polyethylene glycol), a cycloakyl, an aryl, a heterocycloalkyl, a heteroaryl, a carbohydrate, a glycan, a peptidoglycan, a polypeptide, or any combination thereof. In some embodiments, the spacer comprises any one or more of the following units: an amide, an ester, a urea, a carbonate, a carbamate, a disulfide, an amino acid, an amine, an ether, an alkyl, an alkene, an alkyne, a heteroalkyl (e.g., PEG), a cycloakyl, an aryl, a heterocycloalkyl, a heteroaryl, a carbohydrate, a glycan, a peptidoglycan, a polypeptide, or any combination thereof. In some embodiments, the spacer L or L' comprises a solubility enhancer or PK / PD modulating agent W. In some embodiments, the spacer comprises a glycosylated amino acid. In some embodiments, the spacer comprises one or more monosaccharides, disaccharides, polysaccharides, glycans, or peptidoglycans. In some embodiments, the spacer comprises a releasable moiety (e.g., a disulfide bond, an ester, or other moiety that can be cleaved in vivo). In some embodiments, the spacer comprises one or more units of ethylene (e.g., polyethylene), ethylene glycol (e.g., PEG), ethanolamine, ethylenediamine, etc. (e.g., propylene glycol, propanolamine, propylenediamine). In some embodiments, the spacer comprises an oligoethylene, PEG, an alkyl chain, an oligopeptide, a polypeptide, a rigid functional group, a peptidoglycan, an oligoproline, an oligopiperidine, or any combination thereof. In some embodiments, the spacer comprises oligoethylene glycol or PEG. The spacer may comprise oligoethylene glycol.In some embodiments, the spacer comprises PEG. In some embodiments, the spacer comprises an oligopeptide or polypeptide. In some embodiments, the spacer comprises an oligopeptide. In some embodiments, the spacer comprises a polypeptide. In some embodiments, the spacer comprises a peptidoglycan. In some embodiments, the spacer does not comprise a glycan. In some embodiments, the spacer does not comprise a sugar. In some embodiments, the rigid functional group is an oligoproline or oligopiperidine. In some embodiments, the rigid functional group is an oligoproline. In some embodiments, the rigid functional group is an oligopiperidine. In some embodiments, the rigid functional group is an oligophenyl. In some embodiments, the rigid functional group is an oligoalkyne. In some embodiments, the oligoproline or oligopiperidine has from about 2 up to about 50 repeating units (e.g., proline or piperidine), inclusive, from about 2 to about 40, from about 2 to about 30, from about 2 to about 20, from about 2 to about 15, from about 2 to about 10, or from about 2 to about 6 repeating units (e.g., proline or piperidine).

[0165] In some embodiments, the compounds include W groups to enhance the properties of the compounds. 1 is replaced by W, except that one or more G 1 is not W. In some embodiments, one or more G 1 is replaced by W, except that one or more G 1is a folate receptor ligand. In some embodiments, the linker L comprises one or more W groups. In some embodiments, W comprises a solubility enhancer or PK / PD modulating agent. In some embodiments, W comprises polyethylene glycol (PEG), a sugar, a peptide, or a peptidoglycan. In some embodiments, W comprises PEG, a sugar, a peptide, or a peptidoglycan to achieve better solubility and PK / PD properties. In some embodiments, W comprises one or more monosaccharides, disaccharides, peptides, peptidoglycans, and / or serum albumin. In some embodiments, W comprises one or more PEGs, peptides, peptidoglycans, or serum albumins. In some embodiments, W does not comprise a sugar. In some embodiments, W does not comprise a monosaccharide, a disaccharide, or a polysaccharide. In some embodiments, W does not comprise a glycan. In some embodiments, W comprises a glycosylated amino acid. In some embodiments, W comprises a glycosylated cysteine. In some embodiments, W comprises a free carboxylic acid. In some embodiments, W comprises PEG.

[0166] In some embodiments, W comprises one or more monosaccharides, disaccharides, oligosaccharides, polysaccharides, peptides, peptidoglycans, serum albumin, solubility enhancers, PK / PD modifiers, or combinations thereof. In some embodiments, W modulates pharmacological, pharmacokinetic, pharmacodynamic, or physicochemical properties. In some embodiments, W promotes internalization. In some embodiments, W improves water solubility. In some embodiments, W increases plasma protein binding. In some embodiments, W modulates (e.g., reduces) the excretion, elimination, metabolism, stability (e.g., enzymatic stability, plasma stability), distribution, toxicity, or a combination thereof of the compound.

[0167] In some embodiments, the monosaccharides as found in W exist in equilibrium between their linear and cyclic forms. In some embodiments, the monosaccharides are linear. In some embodiments, the monosaccharides are cyclic. In some embodiments, the monosaccharides exist as D-isomers. In some embodiments, the monosaccharides exist as L-isomers. In some embodiments, by way of non-limiting example, W comprises one or more monosaccharides selected from the following: ribose, galactose, mannose, glucose fructose, N-acetylglucosamine, N-acetylmuramic acid, or derivatives thereof (e.g., cyclic or linear forms, methylated derivatives, acetylated derivatives, phosphorylated derivatives, aminated derivatives, oxidized or reduced derivatives, D or L isomers, isotopes, stereoisomers, positional isomers, tautomers, or combinations thereof).

[0168] In some embodiments, the disaccharides, oligosaccharides, or polysaccharides that may be disposed within W contain O-linkages, N-linkages, C-linkages, or combinations thereof. In some embodiments, the disaccharides, oligosaccharides, or polysaccharides contain glycosidic linkages in either the alpha- or beta-orientation. In some embodiments, W comprises an oligosaccharide, polysaccharide, or glycan (e.g., a glycoprotein, glycopeptide, glycolipid, glycogen, proteoglycan, peptidoglycan, etc.).

[0169] In some embodiments, W comprises an amino acid, peptide, polypeptide, or protein. In some embodiments, the amino acid is a natural amino acid (e.g., alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine (Cys), glutamic acid (Glu), glutamine (Gln), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val)). Alternatively, in some embodiments, the amino acid is an unnatural or modified amino acid. W can comprise a sugar or sugar derivative covalently attached to the side chain of an amino acid (e.g., glutamic acid, aspartic acid).

[0170] In some embodiments, W is It may contain glycosylated amino acids such as TIFF2025118775000089.tif16128.

[0171] In some embodiments, the peptide or polypeptide comprises multiple natural and / or unnatural amino acids. In some embodiments, the peptide (or peptidoglycan) has about 2 to about 20 amino acids. In some embodiments, the amino acids, peptides, polypeptides, or proteins (e.g., those located within or comprising W) have a pharmacological or physicochemical effect that improves one or more properties of the compound (e.g., modulating solubility, dissolution, size, permeability, protein binding, target binding, excretion, metabolism, toxicity, distribution, half-life, and / or duration of action). In some embodiments, W is a pharmacokinetic modifier. In some embodiments, the pharmacokinetic modifier is a peptide or protein that can modulate (e.g., enhance) protein binding. In some embodiments, the pharmacokinetic modifier enhances plasma protein binding. In some embodiments, the pharmacokinetic modifier decreases the rate of elimination, excretion, or metabolism. In some embodiments, the pharmacokinetic modifier increases the duration of action of the compound.

[0172] In some embodiments, the linker comprises an albumin ligand. In some embodiments, the albumin ligand is Includes TIFF2025118775000090.tif16128.

[0173] In some embodiments, the linker comprises a dimethylcysteine group. In some embodiments, the dimethylcysteine group is linked to a succinimide. Form TIFF2025118775000091.tif29128.

[0174] Compounds of formula (I) have the formula: TIFF2025118775000092.tif20199, or a pharmaceutically acceptable salt, polymorph, prodrug, solvate, or clathrate thereof, wherein the steroids are dexamethasone, flumethasone, betamethasone, betamethasone 17-valerate, and budesonide, respectively. While the steroid attached to L is shown attached to the carbonyl of the D ring through the hydroxyl group α, the attachment of the steroid described and contemplated herein can be from any suitable portion of the steroid. Furthermore, those skilled in the art will recognize that the folate-targeting ligand and steroid portions of the compound of Formula (I) contain chiral centers. All diastereomers and racemates of the compounds described herein are contemplated.

[0175] Representative compounds Compounds of formula (I) have the formula: Compound 101: TIFF2025118775000093.tif57128; Compound 102: TIFF2025118775000094.tif58130; Compound 103: TIFF2025118775000095.tif62130; Compound 104: TIFF2025118775000096.tif60130; Compound 105: TIFF2025118775000097.tif64130; Compound 106 (Folate-PEG2-hydrolyzed-dimethylcysteine-carbamate-dexamethasone): TIFF2025118775000098.tif40170; Compound 107 (Folate-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Dexamethasone): TIFF2025118775000099.tif66170; Compound 108 (Folate-PEG(CH)-pyrophosphate-dexamethasone): TIFF2025118775000100.tif72134; Compound 109 (Folate-Albumin Binder-PEG(CH)-Dimethylcysteine-Carbamate-Dexamethasone): TIFF2025118775000101.tif57170; Compound 110 (Folate-Albumin Binder-PEG(CH)-Dimethylcysteine-Carbamate-Dexamethasone): TIFF2025118775000102.tif57170; Compound 111 (Folate-PEG(CH)-dimethylcysteine-maleimide-cathepsin B cleavable-dicarbamate-dexamethasone): TIFF2025118775000103.tif85170; Compound 112 (Folate-Albumin Binder-PEG(CH)-Dimethylcysteine-Maleimide-Cathepsin B Cleavable-Dicarbamate-Dexamethasone): TIFF2025118775000104.tif89170; Compound 113 (Folate-Albumin Binder-PEG-Hydrolink-Dimethylcysteine-Maleimide-Cathepsin B Cleavable-(CHCH)-Dicarbamate-Dexamethasone): TIFF2025118775000105.tif61170; Compound 114 (Folate-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Maleimide-Cathepsin B Cleavable-Phosphate-Dexamethasone): TIFF2025118775000106.tif71170; Compound 115 (Folate-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Maleimide-Beta-Glucuronidase Cleavable-Dicarbamate-Dexamethasone): TIFF2025118775000107.tif80170; Compound 116 (Folate-Albumin Binder-PEG(CH)-Dimethylcysteine-Maleimide-Beta-Glucuronidase Cleavable-Dicarbamate-Dexamethasone): TIFF2025118775000108.tif106170; Compound 117 (Pteroyl-dimethylcysteine-cathepsin B cleavable-dicarbamate-dexamethasone): TIFF2025118775000109.tif101170; Compound 118 (Pteroyl-lysine-carbamate-dexamethasone): TIFF2025118775000110.tif56128; Compound 119 (Pteroyl-dimethylcysteine-carbamate-dexamethasone): TIFF2025118775000111.tif39128; Compound 120 (Pteroyl-serine-phosphate-dexamethasone): TIFF2025118775000112.tif41128; Compound 121 (Pteroyl-serine-pyrophosphate-dexamethasone): TIFF2025118775000113.tif50128; Compound 122 (Pteroyl-threonine-phosphate-dexamethasone): TIFF2025118775000114.tif38128; Compound 123 (Pteroyl-threonine-pyrophosphate-dexamethasone): TIFF2025118775000115.tif46128; Compound 124 (Folic Acid-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Flumethasone) TIFF2025118775000116.tif83170; Compound 125 (Fol-Alb-PEG2-hydrolink-dimethylcysteine-maleimide-(CH2)5-cathepsin B-cleavable-dicarbamate-dexamethasone): TIFF2025118775000117.tif90170; Compound 126 (Folic Acid-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Betamethasone) TIFF2025118775000118.tif83170; and Compound 127 (Folic Acid-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Budesonide) TIFF2025118775000119.tif81156 or a pharmaceutically acceptable salt, polymorph, prodrug, solvate or clathrate thereof.

[0176] Pharmaceutical Compositions In certain embodiments, disclosed herein are pharmaceutical compositions comprising one or more compounds described herein (e.g., compounds of Formula (I)) and one or more pharmaceutically acceptable excipients. Excipients are substances that are not active ingredients added to pharmaceutical formulations. Classes of excipients include diluents (e.g., fillers used to increase tablet weight and improve content uniformity, including starch, hydrolyzed starch, and partially pregelatinized starch; other examples of diluents include anhydrous lactose, lactose monohydrate, and sugar alcohols such as sorbitol, xylitol, and mannitol). Such compositions may be specially formulated for administration via one or more of a number of routes, including, but not limited to, buccal, cutaneous, epicutaneous, epidural, infusion, inhalation, intra-arterial, intracardiac, intraventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, pulmonary, rectal via enema or suppository, subcutaneous, subdermal, sublingual, transdermal, and transmucosal. Additionally, administration may be by means of capsules, drops, foams, gels, gums, injections, liquids, patches, pills, porous pouches, powders, tablets, or other suitable means of administration.

[0177] Also contemplated herein are pharmaceutical compositions comprising any of the compounds described herein and at least one pharmaceutically acceptable excipient that is part of a nanoparticle, liposome, or exosome formulation.

[0178] Pharmaceutical excipients provide chemical and / or biological stability and release characteristics, but generally do not provide pharmacological activity to the formulation. Examples of suitable formulations can be found, for example, in Remington, The Science and Practice of Pharmacy, 20th Edition, (Gennaro, AR, Chief Editor), Philadelphia College of Pharmacy and Science, 2000, which is incorporated by reference in its entirety.

[0179] Any suitable pharmaceutically acceptable excipient may be used in the compositions disclosed herein, including any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents that are physiologically compatible.

[0180] In some embodiments, pharmaceutical compositions are suitable for parenteral administration.Alternatively, pharmaceutical compositions are suitable for intravenous, intraperitoneal, intramuscular, sublingual or oral administration.In some embodiments, pharmaceutical compositions are sterile aqueous solutions or dispersions or sterile powders for the immediate preparation of sterile injectable solutions or dispersions.The use of such media and agents for pharmaceutically active substances is well known in the art.Except where any conventional media or agents are incompatible with the active compound, their use in the pharmaceutical compositions disclosed herein is contemplated.A supplementary active compound can also be incorporated into the composition.

[0181] The pharmaceutical composition may be sterile and stable under the conditions of manufacture and storage.The composition may be formulated as a solution, microemulsion, liposome, or other ordered structure suitable for high drug concentration.The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof.The proper fluidity may be maintained, for example, by using a coating such as lecithin, by maintaining the required particle size in the case of dispersion, and by using surfactants.

[0182] In some cases, isotonic agents may be included in the pharmaceutical composition, such as sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride. Prolonged absorption of injectable compositions can be achieved by including agents that delay absorption, such as monostearate salts and gelatin, in the composition. Furthermore, the compounds described herein can be formulated into sustained-release formulations, for example, compositions containing sustained-release polymers. The active compounds may be prepared with carriers that protect the compound from rapid release, such as controlled-release formulations, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, polylactic acid, and polylactic-polyglycolic acid copolymers (PLG), may also be used. Many methods for preparing such formulations are known to those skilled in the art.

[0183] Oral dosage forms are also contemplated herein. Pharmaceutical compositions may be orally administered as capsules (hard or soft), tablets (film-coated, enteric-coated, or uncoated), powders or granules (coated or uncoated), or liquids (solutions or suspensions). Formulations can be conveniently prepared by any method known in the art. Pharmaceutical compositions may contain one or more suitable manufacturing aids or excipients, including fillers, binders, disintegrants, lubricants, diluents, flow agents, buffers, wetting agents, preservatives, colorants, sweeteners, flavoring agents, and pharmaceutically compatible carriers.

[0184] For each of the described embodiments, the compound can be administered in various dosage forms known in the art.Biologically acceptable dosage forms known to those skilled in the art and their combinations are contemplated.Examples of such dosage forms include, but are not limited to, chewable tablets, fast-dissolving tablets, effervescent tablets, reconstitutable powders, elixirs, liquids, solutions, suspensions, emulsions, tablets, multi-layer tablets, bi-layer tablets, capsules, soft gelatin capsules, hard gelatin capsules, caplets, troches, chewable troches, beads, powders, gums, granules, particles, microparticles, dispersible granules, cachets, douches, suppositories, creams, topicals, inhalants, aerosol inhalants, patches, particle inhalants, implants, depot implants, oral administration, injections (including subcutaneous, intramuscular, intravenous, and intradermal), infusions, and combinations thereof.

[0185] Other compounds that may be included by mixing include, for example, medically inert ingredients (e.g., solid and liquid diluents) such as lactose, dextrose, saccharose, cellulose, starch, or calcium phosphate for tablets or capsules, olive oil or ethyl oleate for soft capsules, and water or vegetable oil for suspensions or emulsions; lubricants such as silica, talc, stearic acid, magnesium or calcium stearate, and / or polyethylene glycol; gelling agents such as colloidal clays; thickening agents such as tragacanth gum or sodium alginate, binders such as starch, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, or polyvinylpyrrolidone; disintegrating agents such as starch, alginic acid, alginates, or sodium starch glycolate; effervescent agents; dyes; sweeteners; wetting agents such as lecithin, polysorbate, or lauryl sulfate; and other therapeutically acceptable auxiliary ingredients that are known additives for such formulations, such as humectants, preservatives, buffers, and antioxidants.

[0186] Liquid dispersions for oral administration may be syrups, emulsions, solutions, or suspensions. Syrups may contain, for example, sucrose or sucrose with glycerol and / or mannitol and / or sorbitol as a carrier. Suspensions and emulsions may contain, for example, natural gums, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol.

[0187] The amount of active compound in the compositions of various embodiments may vary depending on factors such as the condition, age, sex, weight, patient history, risk factors, predisposition to the disease, route of administration, and existing treatment regimens (e.g., potential interactions with other drugs). The dosage regimen may be adjusted to provide an optimal response. For example, a single bolus may be administered, or several divided doses may be administered over time, or the dose may be reduced or increased proportionally to the exigencies of the situation. Desirably, a therapeutically effective amount is provided, the amount being sufficient to provide a therapeutic effect in a subject with inflammation, such as inflammation associated with a disease or disorder, or a prophylactic effect in a subject at risk of developing inflammation associated with a disease or disorder.

[0188] As used herein, "dosage unit form" refers to a physically discrete unit suitable as a single dose for a mammalian subject to be treated; each unit contains a predetermined amount of active compound calculated to provide the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit form for use with the methods disclosed herein are determined and directly influenced by the unique properties of the active compound and the specific therapeutic effect to be achieved, as well as the limitations inherent in the technology of formulating such active compounds for the treatment of susceptibility in individuals. In therapeutic use for the treatment of conditions in mammals (e.g., humans) for which the various embodiments of the compounds described herein or their appropriate pharmaceutical compositions are effective, the various embodiments of the compounds described herein can be administered in an effective amount.

[0189] Doses may be administered once, twice, or three times daily, although more frequent dosing intervals are possible. Doses may be administered daily, every two, three, four, five, six, and / or seven days (once a week). In one embodiment, doses may be administered daily for up to 30 days, preferably 7-10 days. In another embodiment, doses may be administered twice daily for 10 days. If a patient requires treatment for a chronic disease or condition, doses may be administered as long as signs and / or symptoms persist. Patients may require "maintenance therapy," in which a dose is given daily for months, years, or even lifelong. Additionally, the compositions disclosed herein may be intended to provide prophylaxis against recurring symptoms. For example, doses may be administered once or twice daily to prevent the onset of symptoms in at-risk patients, particularly for asymptomatic patients.

[0190] The compositions described herein may be administered by any of the following routes: buccal, epicutaneous, epidural, injection, inhalation, intra-arterial, intracardiac, intraventricular, intradermal, intramuscular, intranasal, intraocular, intraperitoneal, intraspinal, intrathecal, intravenous, oral, parenteral, pulmonary, rectal via enema or suppository, subcutaneous, subdermal, sublingual, transdermal, and transmucosal. Preferred routes of administration are buccal and oral. Administration can be local, in which the composition is administered directly to, adjacent to, locally, near, at, around, or near the site of disease, e.g., inflammation, or systemic, in which the composition is given to a patient and distributed throughout the body, thereby reaching the site of disease. Local administration can be to cells, tissues, organs, and / or organ systems that contain and / or are affected by disease and / or where signs and / or symptoms of disease are active or likely to occur. Administration can be local with a local effect, where the composition is applied directly to the location where its action is desired. Administration can be enteral, where the desired effect is systemic (non-local) and the composition is delivered via the digestive tract. Administration can be parenteral, where the desired effect is systemic and the composition is delivered by a route other than the digestive tract.

[0191] Compositions comprising a therapeutically effective amount of one or more compounds of various embodiments described herein (e.g., compounds of Formula (I)) are also contemplated. The compositions are useful in a method for treating inflammation in a subject, comprising administering a therapeutically effective amount of a compound described herein to the subject. A therapeutically effective amount can provide a preventive or therapeutic effect. Inflammation can be treated with reduced or eliminated adverse effects compared to when the steroid is administered without being linked to a folate targeting ligand. Adverse effects of steroids, such as those administered systemically, include sodium (salt) and fluid retention, weight gain, edema, high blood pressure, muscle weakness, delayed wound healing, etc.

[0192] Treatment method In certain aspects, disclosed herein are methods of treating inflammation in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of one or more compounds disclosed herein or a pharmaceutical composition disclosed herein.

[0193] In some embodiments, the inflammation is associated with Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft versus host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor.

[0194] In certain aspects, disclosed herein are methods of transitioning macrophages from M1 to M2 in an individual in need thereof, comprising administering to the individual a therapeutically effective amount of one or more compounds disclosed herein or a pharmaceutical composition disclosed herein.

[0195] In certain embodiments, disclosed herein are methods for treating a disease or disorder involving polarizing macrophages from M1 to M2 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. M1 and M2 macrophage profiles can be used to determine dosage, monitor the effectiveness of treatment (including therapeutic and prophylactic), and adjust dosage treatment regimens. CD86 can be assessed to determine M1 macrophage levels, while CD206 can be assessed to determine M2 macrophage levels.

[0196] In certain embodiments, disclosed herein are methods for treating a disease or disorder involving polarization of tissue-resident macrophages (TRM), also known by other names, from M1 to M2 in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein. Those skilled in the art will recognize that, depending on the organ in which they reside, some TRMs have specific names, such as alveolar macrophages (AM) (lung), microglia (brain), Kupffer cells (liver), renal macrophages (kidney), and osteoclasts (skeletal system).

[0197] In certain aspects, disclosed herein are methods for treating an inflammatory disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition described herein. In some embodiments, the inflammatory disorder is Crohn's disease, lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, or fibrotic disease, graft versus host disease (GVHD), fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, periprosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system, viral infection, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, atherosclerosis, cancer, or tumor.

[0198] In certain aspects, disclosed herein are methods for treating inflammation associated with an autoimmune disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition comprising one or more compounds disclosed herein.

[0199] In certain embodiments, a method for treating inflammation associated with an autoimmune disease in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of budesonide, fludrocortisone, beclomethasone, fluticasone, mometasone, ciclesonide, cortisone, cortivazol, hydrocortisone, 21-hydroxypregnenolone, meprednisone, dexamethasone, triamcinolone, betamethasone, prednisone, prednisolone, and methylprednisolone. In some embodiments, the autoimmune disease is Crohn's disease, lupus, inflammatory bowel disease, Addison's disease, Graves' disease, Sjogren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, asthma, osteoporosis, sarcoidosis, glomerulonephritis, scleroderma, psoriasis, or autoimmune myocarditis.

[0200] In certain aspects, disclosed herein are methods for treating inflammation associated with Crohn's disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0201] In certain embodiments, a method for treating inflammation associated with Crohn's disease in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, budesonide, betamethasone, prednisone, prednisolone, and methylprednisolone.

[0202] In certain aspects, disclosed herein are methods for treating inflammation associated with lupus in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0203] In certain embodiments, a method for treating inflammation associated with lupus in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, hydrocortisone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0204] In certain aspects, disclosed herein are methods for treating inflammation associated with inflammatory bowel disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0205] In certain embodiments, a method for treating inflammation associated with inflammatory bowel disease in a subject in need thereof, comprising administering to a subject a G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, hydrocortisone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0206] In certain aspects, disclosed herein are methods for treating inflammation associated with Addison's disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0207] In certain embodiments, a method for treating inflammation associated with Addison's disease in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, hydrocortisone, fludrocortisone, prednisone, prednisolone, and methylprednisolone.

[0208] In certain aspects, disclosed herein are methods for treating inflammation associated with Graves' disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0209] In certain embodiments, a method for treating inflammation associated with Graves' disease in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of Graves' disease-related inflammatory cytokine, such as erythrocyte sedimentation factor (EGF), a soluble erythrocyte sedimentation factor (SEQ ID NO: 1), a soluble erythrocyte sedimentation factor (SEQ ID NO: 2), a soluble erythrocyte sedimentation factor (SEQ ID NO: 3), a 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0210] In certain aspects, disclosed herein are methods for treating inflammation associated with Sjogren's syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0211] In certain embodiments, a method for treating inflammation associated with Sjogren's syndrome in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0212] In certain aspects, disclosed herein are methods for treating inflammation associated with celiac disease in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0213] In certain embodiments, a method for treating inflammation associated with celiac disease in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0214] In certain aspects, disclosed herein are methods for treating inflammation associated with Hashimoto's thyroiditis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0215] In certain embodiments, a method for treating inflammation associated with Hashimoto's thyroiditis in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0216] In certain aspects, disclosed herein are methods for treating inflammation associated with myasthenia gravis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0217] In certain embodiments, a method for treating inflammation associated with myasthenia gravis in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0218] In certain aspects, disclosed herein are methods for treating inflammation associated with autoimmune vasculitis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0219] In certain embodiments, a method for treating inflammation associated with autoimmune vasculitis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0220] In certain aspects, disclosed herein are methods for treating inflammation associated with reactive arthritis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0221] In certain embodiments, a method for treating inflammation associated with reactive arthritis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0222] In certain aspects, disclosed herein are methods for treating inflammation associated with psoriatic arthritis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0223] In certain embodiments, a method for treating inflammation associated with psoriatic arthritis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0224] In certain aspects, disclosed herein are methods for treating inflammation associated with pernicious anemia in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0225] In certain embodiments, a method for treating inflammation associated with pernicious anemia in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0226] In certain aspects, disclosed herein are methods for treating inflammation associated with ulcerative colitis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0227] In certain embodiments, a method for treating inflammation associated with ulcerative colitis in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein X is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0228] In certain aspects, disclosed herein are methods for treating inflammation associated with rheumatoid arthritis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0229] In certain embodiments, a method for treating inflammation associated with rheumatoid arthritis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein X is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, triamcinolone, beclomethasone, betamethasone, dexamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0230] In certain aspects, disclosed herein are methods for treating inflammation associated with type 1 diabetes in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0231] In certain embodiments, a method for treating inflammation associated with type 1 diabetes in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0232] In certain aspects, disclosed herein are methods for treating inflammation associated with multiple sclerosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0233] In certain embodiments, a method for treating inflammation associated with multiple sclerosis in a subject in need thereof, comprising administering to a subject a G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0234] In certain aspects, disclosed herein are methods for treating inflammation associated with asthma in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0235] In certain embodiments, a method for treating inflammation associated with asthma in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of triamcinolone, fluticasone, budesonide, mometasone, beclomethasone, ciclesonide, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0236] In certain aspects, disclosed herein are methods for treating inflammation associated with osteoporosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0237] In certain embodiments, a method for treating inflammation associated with osteoporosis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0238] In certain aspects, disclosed herein are methods for treating inflammation associated with sarcoidosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0239] In certain embodiments, a method for treating inflammation associated with sarcoidosis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0240] In certain aspects, disclosed herein are methods for treating inflammation associated with glomerulonephritis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0241] In certain embodiments, a method for treating inflammation associated with glomerulonephritis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, budesonide, betamethasone, prednisone, prednisolone, and methylprednisolone.

[0242] In certain aspects, disclosed herein are methods for treating inflammation associated with autoimmune myocarditis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0243] In certain embodiments, a method for treating inflammation associated with autoimmune myocarditis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0244] In certain aspects, disclosed herein are methods for treating inflammation associated with a fibrotic disorder (e.g., a fibrotic disorder resulting from inflammation) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0245] In certain embodiments, a method for treating inflammation associated with a fibrotic disorder (e.g., a fibrotic disorder resulting from inflammation) in a subject in need thereof, comprising administering to a subject a G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition comprising one or more compounds disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, and prednisone.

[0246] In certain aspects, disclosed herein are methods for treating inflammation associated with graft-versus-host disease (GVHD) in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0247] In certain embodiments, a method for treating inflammation associated with graft-versus-host disease (GVHD) in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of GVHD. 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0248] In certain aspects, disclosed herein are methods for treating inflammation associated with fatty liver disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0249] In certain embodiments, a method for treating inflammation associated with fatty liver disease in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0250] In certain aspects, disclosed herein are methods for treating inflammation associated with ischemia-reperfusion injury in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0251] In certain embodiments, a method for treating inflammation associated with ischemia-reperfusion injury in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0252] In certain aspects, disclosed herein are methods for treating inflammation associated with periprosthetic osteolysis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0253] In certain embodiments, a method for treating inflammation associated with periprosthetic osteolysis in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0254] In certain aspects, disclosed herein are methods for treating inflammation associated with scleroderma in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition comprising one or more compounds disclosed herein.

[0255] In certain embodiments, a method for treating inflammation associated with scleroderma in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, budesonide, betamethasone, triamcinolone, prednisone, prednisolone, and methylprednisolone.

[0256] In certain aspects, disclosed herein are methods for treating inflammation associated with psoriasis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0257] In certain embodiments, a method for treating inflammation associated with psoriasis in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of budesonide, betamethasone, triamcinolone, prednisone, prednisolone, methylprednisolone, hydrocortisone, dexamethasone, hydrocortisone 17-valerate, diflorasone, meprednisone, halobetacol, tixocortol, amcinonide, desonide, fluocinolone acetonide, fluocinonide, halcinonide, beclomethasone, and halomethasone.

[0258] In certain aspects, disclosed herein are methods for treating inflammation associated with spinal cord injury in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0259] In certain embodiments, a method for treating inflammation associated with spinal cord injury in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0260] In certain aspects, disclosed herein are methods for treating inflammation of the central nervous system in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0261] In certain embodiments, a method for treating inflammation of the central nervous system in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0262] In certain aspects, disclosed herein are methods for treating inflammation associated with a viral infection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0263] In certain embodiments, a method for treating inflammation associated with a viral infection in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0264] In certain aspects, disclosed herein are methods for treating inflammation associated with influenza in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0265] In certain embodiments, a method for treating inflammation associated with influenza in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0266] In certain aspects, disclosed herein are methods for treating inflammation associated with SARS-CoV-2 (COVID-19) infection in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0267] In certain embodiments, a method for treating inflammation associated with SARS-CoV-2 (COVID-19) infection in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein X is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, and prednisone, prednisolone, and methylprednisolone.

[0268] In certain aspects, disclosed herein are methods for treating inflammation associated with cytokine storm syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0269] In certain embodiments, a method for treating inflammation associated with cytokine storm syndrome in a subject in need thereof comprises administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of hydrocortisone, dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0270] In certain aspects, disclosed herein are methods for treating inflammation associated with damage to bone in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0271] In certain embodiments, a method for treating inflammation associated with injury to bone in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein X is a radical of a steroid selected from the group consisting of cortisone, cortivazol, hydrocortisone, 21-hydroxypregnenolone, meprednisone, dexamethasone, triamcinolone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0272] In certain aspects, disclosed herein are methods for treating inflammation associated with an inflammatory brain disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0273] In certain embodiments, a method for treating inflammation associated with an inflammatory brain disease in a subject in need thereof, comprising administering to a subject a G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of dexamethasone, betamethasone, budesonide, prednisone, prednisolone, and methylprednisolone.

[0274] In certain aspects, disclosed herein are methods for treating inflammation associated with atherosclerosis in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0275] In certain embodiments, a method for treating inflammation associated with atherosclerosis in a subject in need thereof comprises administering to a subject a G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of fluticasone, budesonide, beclomethasone, ciclesonide, dexamethasone, betamethasone, prednisone, prednisolone, and methylprednisolone.

[0276] In certain aspects, disclosed herein are methods for treating inflammation associated with a tumor in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0277] In certain embodiments, a method for treating inflammation associated with a tumor in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone.

[0278] In certain aspects, disclosed herein are methods for treating inflammation associated with cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein.

[0279] In certain embodiments, a method for treating inflammation associated with cancer in a subject in need thereof, comprising administering to a subject a therapeutically effective amount of G 2 Disclosed herein are methods that include administering to a subject a therapeutically effective amount of a compound disclosed herein or a pharmaceutical composition disclosed herein, wherein is a radical of a steroid selected from the group consisting of cortisone, hydrocortisone, clobetasol, dexamethasone, betamethasone, budesonide, meprednisone, prednisone, prednisolone, and methylprednisolone.

[0280] In some embodiments, a therapeutically effective amount is one that can treat or alleviate disease, signs or symptoms of disease, with a reasonable benefit / risk ratio applicable to any medical treatment.However, it should be understood that the total daily use amount of the compounds and compositions described herein can be determined by the attending physician within the scope of sound medical judgment.The specific therapeutically effective dose level for any particular patient will vary depending on various factors, including the condition being treated and the severity of the condition; the activity of the specific compound being used; the specific composition being used; the patient's age, weight, general health, sex and diet; the administration time, administration route and excretion rate of the specific compound being used; the duration of treatment; drugs used in combination with or simultaneously with the specific compound being used; and similar factors well known to researchers, veterinarians, physicians or other clinicians.It is also recognized that a therapeutically effective amount can be selected with reference to any toxicity or other undesirable adverse effects that may occur when administering one or more of the compounds described herein.

[0281] It will be appreciated by those skilled in the relevant art that other suitable modifications and adaptations to the compositions and methods described herein will be readily apparent from the disclosure contained herein in light of information known to those skilled in the art and may be made without departing from the scope of the disclosure. While the present disclosure has been described in detail above, it will be more clearly understood by reference to the following examples, which are for illustrative purposes only and are not intended to be limiting of the disclosure. [Example]

[0282] The present disclosure can be better understood by reference to the following examples, which are presented by way of illustration.

[0283] All LC-MS data were acquired using a 0-100% gradient method utilizing a C18 column with 20 mM ammonium bicarbonate pH 7 and acetonitrile.

[0284] Values expressed in range format should be interpreted flexibly to include not only the numerical values explicitly stated as the limits of the range, but also all individual numerical values or subranges within that range, as if each numerical value and subrange were explicitly stated. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be interpreted to include not only about 0.1% to about 5%, but also individual values (e.g., 1%, 2%, 3%, and 4%) and subranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within the stated range. The phrase "about X to Y" has the same meaning as "about X to about Y" unless otherwise specified. Similarly, the phrase "about X, Y, or about Z" has the same meaning as "about X, about Y, or about Z" unless otherwise specified.

[0285] Example 1 This example describes the synthesis of a compound containing folic acid as a ligand, a linker, and dexamethasone as a steroid.

[0286] Figure 1 shows the chemical structure of "Drug Compound," also known as Compound 101. The drug compound is composed, from left to right, of folic acid as the ligand. The folic acid ligand is covalently attached to a hydrophilic linker consisting of alternating unnatural amino acids D-glutamic acid, diaminopropionic acid (Dap), and the natural amino acid cysteine. The cysteine is then connected to dexamethasone via a disulfide self-immolative linker. The structure shown is also referred to as Compound 101 or folate-hydrolyzed cysteine-carbonate-dexamethasone.

[0287] The compound shown in Figure 1 can be synthesized via the scheme shown in Figure 4 by reacting activated dexamethasone, synthesized via the scheme shown in Figure 2, with the compound shown in Figure 3. Briefly, one equivalent of 6-chloro-1H-benzo[d][1,2,3]triazol-1-yl(2-(pyridin-2-yldisulfanayl)ethyl) carboxylic acid is reacted with one equivalent of dexamethasone (Cayman Chemical Co., Ann Arbor, MI, USA) in dimethyl sulfoxide (DMSO) at room temperature overnight. The product was purified by reverse-phase high-pressure liquid chromatography (RPHPLC) and confirmed by liquid chromatography-mass spectrometry (LC-MS).

[0288] Example 2 The compound in Figure 3 was synthesized via solid-phase peptide synthesis (SPPS) using a standard SPPS protocol with N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (HBTU) and coupling reagents. The product was purified by RP-HPLC and confirmed by LC-MS.

[0289] The product of the reaction scheme in Figure 2 is then reacted with the compound in Figure 3 to give the compound in Figure 4. Briefly, 1 equivalent of the compound in Figure 3 was dissolved in ammonium acetate buffer, pH 6.5. Activated dexamethasone (1 equivalent) was then dissolved in tetrahydrofuran (THF). THF was added to the buffer containing the compound in Figure 3 and stirred overnight at room temperature. The reaction was then purified by RPHPLC and confirmed by LC-MS. The LC-MS trace of the product, Compound 101, in Figure 4 is shown in Figure 16.

[0290] Example 3 General solid-phase peptide synthesis of folate-linker compounds As shown in Scheme 6, 2-chlorotrityl chloride resin was swollen with anhydrous dichloromethane and then loaded with a solution containing 2 equivalents of Fmoc-S-trityl-L-penicillamine (Fmoc-Pen(trt)-OH, also known as Fmoc-dimethylCys-OH) and 10 equivalents of either N-methylmorpholine (NMM) or DIPEA. This was stirred in a solid-phase peptide synthesis vessel under argon at room temperature for 2 hours, after which the resin was capped with the addition of 10% total methanol over 20 minutes. The resin was then washed with DCM / DMF / DCM, dried, and loaded into an automated solid-phase peptide synthesizer. The remaining portion of the molecule was synthesized via solid-phase peptide synthesis (SPPS) using standard SPPS protocols with N,N,N',N'-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (HBTU) / 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) as coupling reagents. 10 -trifluoroacetylation (N 10 -TFA) pteroic acid was finally coupled using the standard SPPS protocol described above, followed by N using 50% ammonium hydroxide in DMF. 10 -TFA deprotection was performed. The product was cleaved from the resin with TFA / TIPS / TCEP-HCl, precipitated in diethyl ether, centrifuged, washed three times with diethyl ether and confirmed by LC-MS. TIFF2025118775000120.tif140152 Scheme 6 - Synthesis scheme of steroid-carbamate-pyridyl disulfide compounds

[0291] Example 4 General procedure for the synthesis of steroid-carbamate-pyridyl disulfide compounds: TIFF2025118775000121.tif147135 Scheme 7 - Synthesis scheme of steroid-carbamate-pyridyl disulfide compounds

[0292] As shown in Scheme 7, a steroid (1 equivalent, dexamethasone, betamethasone, flumethasone, or budesonide) and p-nitrophenyl chloroformate (1 equivalent) were charged to a capped vial containing a magnetic stir bar. Next, the addition of either chloroform or dichloromethane resulted in a final steroid concentration of 0.1 M. Finally, approximately 25 equivalents of pyridine were added to the stirred solution, and the reaction was monitored at room temperature by LC-MS. Approximately 1.4 equivalents of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt was then mixed with chloroform or dichloromethane and approximately 4.3 equivalents of diisopropylethylamine (DIPEA) in a separate vessel, and monitoring continued by LC-MS. Upon completion, the reaction was purified by flash chromatography utilizing either a 0-100% hexane / ethyl acetate or 0-20% dichloromethane / methanol gradient. The product was confirmed by LC-MS.

[0293] Synthesis of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt 25.6 mmol of methoxycarbonylsulfenyl chloride in 10 mL of acetonitrile was cooled to 0°C, followed by the addition of 25.6 mmol of 2-aminoethanol hydrochloride. 23 mmol of 2-mercaptopyridine in 15 mL of methanol was then added, and the mixture was refluxed for 2 hours. The vessel was then cooled to 0°C for 1 hour, and the resulting precipitate was collected by filtration. (Yield: 2.5660 g)

[0294] Synthesis of Dex-carbamate-pyridyl disulfide A capped vial containing a magnetic stir bar was charged with 392 mg of dexamethasone and 230 mg of p-nitrophenyl chloroformate. The solid was dissolved in 10 mL of chloroform, followed by the addition of 1 mL of pyridine. The reaction was carried out at room temperature and monitored by LC-MS. 320 mg of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt was then mixed with 2.5 mL of chloroform and 752 μL of diisopropylethylamine (DIPEA) in a separate container, and monitoring continued by LC-MS. Upon completion, the reaction was purified by flash chromatography using a 0-20% dichloromethane / methanol gradient. The product was confirmed by LC-MS. (Yield 460.0 mg)

[0295] Synthesis of beta-carbamate-pyridyl disulfides A capped vial containing a magnetic stir bar was charged with 10.0 mg of betamethasone and 5.4 mg of p-nitrophenyl chloroformate. Next, 200 μL of dichloromethane was added, followed by 40 μL of pyridine, to the stirred solution, and the reaction was monitored at room temperature by LC-MS. 5.7 mg of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt was then mixed with 500 μL of dichloromethane and 13.3 μL of diisopropylethylamine (DIPEA) in a separate container, and monitoring continued by LC-MS. Upon completion, the reaction was purified by flash chromatography utilizing a 0-20% dichloromethane / methanol gradient followed by 0-100% hexanes / ethyl acetate. The product was confirmed by LC-MS. (Yield: 6.2 mg)

[0296] Synthesis of Flu-carbamate-pyridyl disulfides A capped vial containing a magnetic stir bar was charged with 10.6 mg of flumethasone and 5.4 mg of p-nitrophenyl chloroformate. 213 μL of dichloromethane followed by 42.7 μL of pyridine was added, and the reaction was monitored at room temperature by LC-MS. 5.75 mg of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt was then mixed with 500 μL of dichloromethane and 12.5 μL of diisopropylethylamine (DIPEA) in a separate container, and monitoring continued by LC-MS. Upon completion, the reaction was purified by flash chromatography using a 0-20% dichloromethane / methanol gradient. The product was confirmed by LC-MS. (Yield 5.8 mg)

[0297] Synthesis of Bud-carbamate-pyridyl disulfide A capped vial containing a magnetic stir bar was charged with 25 mg of budesonide and 12.3 mg of p-nitrophenyl chloroformate. 500 μL of chloroform and 100 μL of pyridine were added to the stirred solution, and the reaction was monitored at room temperature by LC-MS. 12.9 mg of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt was then mixed with 500 μL of chloroform and 30.3 μL of diisopropylethylamine (DIPEA) in a separate container and monitored by LC-MS. Upon completion, the reaction was purified by flash chromatography using a 0-20% dichloromethane / methanol gradient. The product was confirmed by LC-MS. (Yield 30.6 mg)

[0298] Example 5 General synthesis of folate-albumin binder-PEG2-hydrolinked-dimethylcysteine-carbamate-steroid compounds TIFF2025118775000122.tif141142Scheme 8 Final dimethyl Cys-maleimide coupling of compounds

[0299] General synthesis of folate-albumin binder-PEG2-hydrolinked-dimethylcysteine-carbamate-steroid compounds As shown in Scheme 8, 1-2 equivalents of Fol-Alb-PEG2-hydrolinked-dimethylCys were dissolved in 20 mM ammonium acetate buffer (pH 5-7) and vigorously stirred. One equivalent of steroid-carbamate-pyridyl disulfide dissolved in THF or DMF (an equal volume to the buffer) was then quickly added and monitored by LC-MS until completion. The reaction was then purified by RP-HPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS.

[0300] Compound 107 (Folic Acid-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Dexamethasone) 60 mg of Fol-Alb-PEG2-hydrolink-dimethylCys was dissolved in 1 mL of 20 mM ammonium acetate buffer (pH 6) and vigorously stirred. 19.2 mg of Dex-carbamate-pyridyl disulfide dissolved in 1 mL of THF was then quickly added and monitored by LC-MS until completion. The reaction was then purified by RP-HPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS. (Compound 107; yield 15.8 mg). The LC-MS is shown in Figure 18.

[0301] Compound 124 (Folic Acid-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Flumethasone) 26 mg of Fol-Alb-PEG2-hydrolink-dimethylCys was dissolved in 250 μL of 20 mM ammonium acetate buffer (pH 6.19) and vigorously stirred. 5.8 mg of Flu-carbamate-pyridyl disulfide dissolved in 250 μL of THF was then quickly added and monitored by LC-MS until completion. The reaction was then purified by RP-HPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS. (Compound 124; yield 1 mg) LC-MS is shown in Figure 21.

[0302] Folic acid-albumin binder-PEG2-hydrolyzed dimethylcysteine-carbamate-betamethasone (compound 126) 28.7 mg of Fol-Alb-PEG2-hydrolink-dimethylCys was dissolved in 0.250 mL of 20 mM ammonium acetate buffer (pH 6) and vigorously stirred. 6.2 mg of betamethasone-carbamate-pyridyl disulfide dissolved in 0.250 mL of THF was then quickly added and monitored by LC-MS until completion. The reaction was then purified by RP-HPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS. (Compound 126; yield 2.6 mg). The LC-MS is shown in Figure 22.

[0303] Folic acid-albumin binder-PEG2-hydrolyzed dimethyl Cys-carbamate-budesonide (compound 127) 21.8 mg of Fol-Alb-PEG2-hydrolink-dimethylCys was dissolved in 0.250 mL of 20 mM ammonium acetate buffer (pH 6) and vigorously stirred. 5 mg of betamethasone-carbamate-pyridyl disulfide dissolved in 0.250 mL of THF was then quickly added and monitored by LC-MS until completion. The reaction was then purified by RP-HPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS. (Compound 127; yield 3.0 mg). The LC-MS is shown in Figure 23.

[0304] Synthesis of Compound 106 (Folic Acid-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Dexamethasone) TIFF2025118775000123.tif126156 Scheme 9: Synthesis scheme for folic acid-PEG2-hydrolyzed-dimethylcysteine-carbamate-dexamethasone (compound 106)

[0305] 39.2 mg of dexamethasone and 23.0 mg of p-nitrophenyl chloroformate were dissolved in 1 mL of chloroform and 100 μL of pyridine. The reaction was run for 2 h, washed with water, diluted with HCl, water, dried over magnesium sulfate, filtered, vacuum dried, and triturated with diethyl ether. (Yield: 10.3 mg) The product was dissolved in 150 μL of DMF, followed by 7.3 μL of DIPEA and 4.7 mg of 2-(pyridin-2-yldisulfaneyl)ethan-1-amine acid chloride salt. The reaction was monitored by LC-MS, and upon completion, the crude reaction mixture was precipitated with diethyl ether, dried, dissolved in 300 μL of DMF, and 300 μL of 20 mM ammonium acetate buffer (pH 6) containing 20 mg of Fol-mPEG2-hydrolink-dimethylCys was quickly added. The reaction was monitored by LC-MS until completion. The reaction was then purified by RPHPLC using 20 mM ammonium acetate buffer / acetonitrile and confirmed by LC-MS. The LC-MS of compound 106 is shown in Figure 17.

[0306] Example 6 Synthesis of Compound 125 (Folate-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Cathepsin B Cleavable-Dicarbamate-Dexamethasone) TIFF2025118775000124.tif166129 Scheme 10 - Synthetic scheme for maleimide-cathepsin B cleavable-dicarbamate-Dex

[0307] Synthesis of Compound 125 (Folate-Albumin Binder-PEG-Hydrolink-Dimethylcysteine-Maleimide-(CH)-Cathepsin B Cleavable-Dicarbamate-Dexamethasone) As shown in Scheme 10, a capped vial containing a magnetic stir bar was charged with dexamethasone (1 equivalent) and p-nitrophenyl chloroformate (1 equivalent). Next, the addition of either chloroform or dichloromethane resulted in a final steroid concentration of 0.1 M. Finally, approximately 25 equivalents of pyridine was added to the stirred solution, and the reaction was monitored at room temperature by LC-MS. Approximately 1 equivalent of (S)-2-((methylamino)methyl)pyrrolidine-1-carboxylate tert-butyl was then mixed with chloroform or dichloromethane and diisopropylethylamine (DIPEA) in a separate container. Upon completion or reaction, as monitored by LC-MS, the product was purified by flash chromatography utilizing a 0-20% dichloromethane / methanol gradient to give tert-butyl (S)-2-(((2-((8S,9R,10S,11S,13S,14S,16R,17R)-9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethoxy)carbonyl)(methyl)amino)methyl)pyrrolidine-1-carboxylate.

[0308] Next, 12.9 mg of tert-butyl (S)-2-(((2-((8S,9R,10S,11S,13S,14S,16R,17R)-9-fluoro-11,17-dihydroxy-10,13,16-trimethyl-3-oxo-6,7,8,9,10,11,12,13,14,15,16,17-dodecahydro-3H-cyclopenta[a]phenanthren-17-yl)-2-oxoethoxy)carbonyl)(methyl)amino)methyl)pyrrolidine-1-carboxylate was treated with a 1:1 mixture of trifluoroacetic acid (TFA) and dichloromethane (DCM). Removal of the Boc protecting group was monitored and confirmed by LC-MS. The TFA / DCM mixture was evaporated under vacuum and the resulting oil was dissolved in 200 uL of THF plus 600 uL of DMF followed by 28.3 uL of DIPEA to which was added 15 mg of commercially supplied (4-nitrophenyl) 4-((R)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)hexanamido)-3-methylbutanamido)-5-ureidopentanamido)benzyl (4-nitrophenyl)carbonate.

[0309] This intermediate was confirmed by LC-MS, and the reaction mixture was then added to 37.9 mg of Fol-Alb-PEG2-hydrolink-dimethylCys in 800 μL of 20 mM ammonium acetate buffer, pH 5.18, under vigorously stirring, as shown in Scheme 10. The product was then purified by RPHPLC using 20 mM ammonium acetate buffer, pH 7 / acetonitrile, and confirmed by LC-MS. (Compound 125; yield 6.74 mg). The LC-MS is shown in Figure 19. TIFF2025118775000125.tif128156 Scheme 11 - Synthesis of Compound 125 (Fol-Alb-PEG2-Hydrolink-Dimethylcysteine-Maleimide-(CH2)5-Cathepsin B Cleavable-Dicarbamate-Dexamethasone)

[0310] Example 8 Synthesis of folate-PEG-pyrophosphate-dexamethasone TIFF2025118775000126.tif74170 Scheme 12 - Synthesis scheme for compound 108 (Folate-PEG12(CH2)-pyrophosphate-dexamethasone)

[0311] Synthesis of 21-dexamethasone phosphate As shown in Scheme 12, 1 g of dexamethasone was added to a 50 mL round-bottom flask followed by 5 mL of anhydrous THF. The reaction was then cooled to -40°C in a dry ice / acetonitrile cold bath. 1.06 mL of diphosphoryl chloride was added, and the reaction was stirred at -40°C for 1 hour. The reaction was quenched with water, titrated to approximately pH 8 with saturated sodium bicarbonate, and then adjusted to approximately pH 2 with 1 M hydrochloric acid. The resulting precipitate was extracted with ethyl acetate, washed with acidified brine, dried over sodium sulfate, and then evaporated in vacuo and used in the next step without further purification. (Yield 1.0231 g)

[0312] Synthesis of Fmoc-phosphate 1 g of (9H-fluoren-9-yl)methyl (2-hydroxyethyl)carbamate followed by 6.8 mL of anhydrous THF was added to a 50 mL round-bottom flask. The reaction was then cooled to -40 °C in a dry ice / acetonitrile cold bath. 1.2 mL of diphosphoryl chloride was added, and the reaction was stirred at -40 °C for 1 hour. The reaction was quenched with water, titrated to approximately pH 8 with saturated sodium bicarbonate, and then adjusted to approximately pH 2 with 1 M hydrochloric acid. The resulting precipitate was extracted with ethyl acetate, washed with acidified brine, dried over sodium sulfate, and then concentrated in vacuo and used in the next step without further purification. (Yield: 1.2870 g)

[0313] Synthesis of Fmoc-Pyro-Dex 50 mg of Fmoc-phosphate was dissolved in 0.4 mL of DMF, followed by the addition of 0.02 mL of trimethylamine and 56.7 mg of carbonyldiimidizole (CDI) over approximately 30 minutes. Then, 63.3 mg of 21-dexamethasone phosphate was added, followed by 150 mg of anhydrous zinc(II) chloride. The reaction was monitored by LC-MS, and upon completion, acetonitrile and 20 mM ammonium acetate buffer, pH 7, were added and lyophilized. The resulting residue was then dissolved in DMSO and purified by RP-HPLC. (Yield: 27.06 mg)

[0314] Synthesis of Fmoc-PEG12-Pyro-Dex 30.5 mg of Fmoc-Pyro-Dex was first dissolved in 10 mL of 6% diethylamine (DEA) in DCM, and Fmoc deprotection was monitored by LC-MS. The DEA / DCM mixture was then removed under vacuum. A solution of 31.3 mg of Fmoc-PEG12-acid was activated with 14.1 mg of HATU, 14.1 mg of Cl-HOBt, 900 μL of DMF, and 40.9 μL of NMM. This mixture was then added to the Fmoc-deprotected Pyro-Dex residue, and upon completion, as monitored by LC-MS, the product was purified by RP HPLC in 20 mM ammonium acetate buffer, pH 7, in acetonitrile, and confirmed by LC-MS.

[0315] Synthesis of folate-NHS 100 mg of folic acid was dissolved in 10 mL of DMSO, followed by the addition of 31.3 mg of NHS and 46.7 mg of N,N'-dicyclohexylcarbodiimide. The mixture was stirred overnight at room temperature. The product was then precipitated into ethyl acetate, filtered by vacuum filtration, and washed three times with diethyl ether. The resulting solid was used without further purification. (Yield: 103.4 mg)

[0316] Synthesis of Compound 108 (Folic Acid-PEG(CH)-Pyrophosphate-Dexamethasone) The Fmoc-PEG12-Pyro-Dex was then deprotected with DEA / DCM and then dried under vacuum. 2.2 mg of the resulting deprotected compound was mixed with 2 mg of folic acid-NHS in 250 μL of DMSO and 1 μL of DIPEA. The reaction was monitored by LC-MS and, upon completion, purified by RPHPLC and confirmed by LC-MS. (Compound 108; yield 1.93 mg). The LC-MS is shown in Figure 20.

[0317] Example 9 Figure 6 shows an example of compound 101 in flow cytometry of macrophages gated on F4 / 80 to show CD206 expression (a marker for M2, anti-inflammatory macrophages). Briefly, 1.5 mL of 3% thioglycollate was injected intraperitoneally into 12-week-old ND4 mice. Three days later, macrophages were isolated by intraperitoneal lavage, plated in 12-well dishes, and incubated at 37°C / 5% CO2. After 24 hours, the medium was replaced with medium containing either 20 nmol of the compound shown in Figure 1, or 20 nmol of the compound shown in Figure 1 plus a 100-fold excess of folate-glucosamine (to demonstrate receptor-specific binding), or untreated control, or 10 nmol of free dexamethasone (to demonstrate the effect of free drug), and incubated at 37°C / 5% CO2 for the indicated time. After 3 days, the cells were isolated and incubated with the corresponding fluorescently labeled antibodies in phosphate-buffered saline (PBS) containing 1% fetal bovine serum (FBS) for 1 hour at 4°C. The samples were then washed three times with ice-cold PBS containing 1% FBS and analyzed by flow cytometry.

[0318] Increased CD206 expression in the target group compared with the competitor group indicated receptor-specific uptake of the drug in macrophages.

[0319] Example 10 FIG. 7 is another example of Compound 101 in flow cytometry of macrophages gated on F4 / 80 to show CD206 expression by control.

[0320] Example 11 Figure 8 shows an example of Compound 101 in macrophage flow cytometry to demonstrate the expression of CD86 compared to control. CD86, a marker for M1, proinflammatory macrophages, showed no observable differences between the various groups treated with compound, competitor, or free dexamethasone over 24 hours, indicating that the steroid had no effect on M1 CD86 expression. This indicates that dexamethasone has no effect on proinflammatory macrophages.

[0321] Example 12 Mechanism research using peritonitis Peritonitis was induced in either BALB / C or Swiss Webster mice maintained on a folic acid-free diet for at least 3 weeks by intraperitoneal injection of 1.5 mL of 3% thioglycollate. 48 hours after induction, the mice were treated with an intravenous injection of 10 nmol of compound 107. After a further 48 hours, the mice were euthanized, and peritoneal lavage was performed using phosphate-buffered saline (PBS) containing 2% fetal bovine serum (FBS). Cells were passed through a 70 μm cell filter to remove debris and then counted using a hemocytometer. Approximately 1 million cells were preincubated in 100 μL of 2% FBS-containing PBS on ice for 1 hour in anti-CD16 / 32 (to block endogenous Fc receptor binding). The cells were then incubated with at least one of the following fluorescently labeled antibodies for cell surface markers: PE-F4 / 80 or PE / Cy7-F4 / 80 (mouse macrophages), FITC-CD4 (T helper cells), eFluor780-CD8 (cytotoxic T cells), and PerCP / Cy5.5-Ly6G (neutrophils) for 1 hour on ice according to the manufacturer's recommended procedure. The cells were washed twice with 1 mL of ice-cold 2% FBS-containing PBS and then suspended in 2% FBS-containing PBS. Controls included unstained cells as a negative control and a positive control using one drop of compensation beads per 1 μL of dye antibody, which was incubated on ice for 1 hour and washed twice with 1 mL of 2% FBS-containing PBS. All samples were centrifuged at 400 × g for 10 minutes between washes, transfers, and after incubation to remove the supernatant.

[0322] Samples and appropriate controls were analyzed on an Attune NxT Acoustic Focusing Cytometer, collecting 10,000–30,000 events per sample using laser lines BL1-A, BL2-A, BL3-A, and YL4-A for FITC-CD4, PE-F4 / 80, PerCP / Cy5.5-Ly6G, and eFluor780-CD8 or PE / Cy7-F4 / 80, respectively. Flow cytometry data analysis first involved observing cells in the experiment using forward and side scatter (FSC and SSC, respectively). All observed cells were gated (R1) and analyzed for the relative abundance of F4 / 80 (murine macrophages), CD4 (T helper cells), CD8 (cytotoxic T cells), and Ly6G (neutrophils) cells within the population. Values from four-quadrant gating based on negative controls were then presented in bar graph format.

[0323] Treatment with Compound 106 (Folic Acid-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Dexamethasone) Figure 9 shows F4 / 80 cytometry results for the first set of samples in the top row and the second set of samples in the bottom row. The left side of each row is a graph showing the results for cells treated with Compound 106, the middle of each row is a graph showing the results for untreated cells, and the right side of each row is a graph showing the results for vehicle-treated cells. Figure 10 is a bar graph summarizing the results from Figure 9. As shown in Figures 9 and 10, the percentage of F4 / 80 macrophages is lower in the treated groups compared to untreated or vehicle. This indicates that macrophages are one of the primary mediators of inflammation, and therefore fewer macrophages are present in the peritoneal cavity after treatment with Compound 106, resulting in less inflammation.

[0324] Treatment with Compound 107 (Folate-Albumin Binder-PEG2-Hydrolink-Dimethylcysteine-Carbamate-Dexamethasone): Figure 11 shows the cytometry results for untreated or compound 107-treated cells for four different immune cell types, and Figure 12 is a bar graph summarizing the results of Figure 11. As shown in Figures 11 and 12, the percentages of PE-F4 / 80 (mouse macrophages), FITC-CD4 (T helper cells), eFluor780-CD8 (cytotoxic T cells), and PerCP / Cy5.5-Ly6G (neutrophils) are lower in the treated groups compared to untreated or vehicle. The immune cells specifically identified above represent the majority of cells involved in inflammation, and fewer are present in the peritoneal cavity after treatment with compound 107.

[0325] Treatment with Compound 108 (Folate-PEG(CH)-pyrophosphate-dexamethasone): Figure 13 shows the cytometry results for untreated or Compound 108-treated cells for four different immune cell types, and Figure 14 is a bar graph summarizing the results of Figure 13. As can be seen in the upper and corresponding lower bar graphs, the percentages of PE-F4 / 80 (mouse macrophages), FITC-CD4 (T helper cells), eFluor780-CD8 (cytotoxic T cells), and PerCP / Cy5.5-Ly6G (neutrophils) are lower in the treated groups compared to untreated or vehicle. The immune cells specifically shown above represent the majority of cells involved in inflammation, and fewer are present in the peritoneal cavity after treatment with Compound 108.

[0326] Example 13 ulcerative colitis Eight- to 12-week-old C57BL / 6 mice were maintained on a folic acid-free diet for at least 3 weeks, and dextran sulfate sodium (DSS) was added to their drinking water at a final concentration of 2.5% for 6 days. On days 3 and 4, mice received a single intravenous injection of 10 nmol of compound 107 in 2% DMSO-containing PBS. On day 6, mice were euthanized by CO2 asphyxiation followed by cervical dislocation. The colons were then excised and prepared for histological examination as previously described (J Vis Exp. 2012;(60):3678). Briefly, the colons were excised, gently removed of feces, and washed with PBS. The colons were fixed in 10% buffered formalin for 24 hours and then transferred to 70% ethanol. These samples were submitted to the Purdue Veterinary Pathology lab, where they were embedded in paraffin, sectioned, and stained with hematoxylin and eosin. The results are shown in Figure 15, with an image of the untreated colon on the left and the treated colon on the right. Note that the treated group had more crypt structures, fewer infiltrating immune cells, and a higher number of goblet cells, indicating a healthier colon.

[0327] Those skilled in the art will recognize that numerous variations can be made to the specific embodiments described above, and the embodiments should not be limited to the specific limitations set forth. Other embodiments may be possible.

Claims

1. Compounds of formula (I), or pharmaceutically acceptable salts thereof: During the ceremony G 1 It is a folate radical; L is a linker; and G 2 It is the radical of dexamethasone, The folic acid radical is represented by the following formula: It has the structure (wherein the formula, the asterisk indicates the bond site of the carbonyl carbon to the linker L), and The linker is an amide, a glucuronide, and the following formula: (In the formula, x'' is an integer between 0 and 10.) Includes a group selected from groups having the structure of [this structure].

2. a. The compound according to claim 1, b. Pharmaceutically acceptable excipients and A pharmaceutical composition containing [the specified substance].

3. The pharmaceutical composition according to claim 2, wherein the pharmaceutically acceptable excipient is part of a nanoparticle, liposome formulation, or exosome formulation.

4. A pharmaceutical composition for treating ulcerative colitis in a subject for which such treatment is necessary, comprising a therapeutically effective amount of the compound according to claim 1.

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