Compounds targeting fibroblast-activating proteins, and methods for using the same.

Abstract

Fibroblast activation protein (FAP) targeting compounds; methods for imaging cancer and fibrosis; and methods for treating fibrosis, inflammatory diseases / disorders and cancer.

Description

[Technical Field]

[0001] Priority This patent application relates to (a) U.S. Provisional Patent Application No. 63 / 200,593 filed on 16 March 2021; (b) U.S. Provisional Patent Application No. 63 / 165,563 filed on 24 March 2021; (c) U.S. Provisional Patent Application No. 63 / 200,592 filed on 16 March 2021; (d) U.S. Provisional Patent Application No. 63 / 165,550 filed on 24 March 2021; (e) U.S. Provisional Patent Application No. 63 / 161,835 filed on 16 March 2021; and (f) U.S. Provisional Patent Application No. 63 / 165,583 filed on 24 March 2021, and claims the benefit of their priority. The entire content of each of the above-mentioned applications is incorporated herein by reference. [Background technology]

[0002] Tumor survival and growth may, non-limitingly, depend on the tumor microenvironment (TME), including the percentage of tumor stroma (TSP). High TSP may be associated with even poorer patient long-term survival compared to low TSP (>50% vs. ≤50%, respectively). TSP may also be an important prognostic factor for tumor recurrence, growth, and metastasis.

[0003] Apart from cancer cells, tumors (and TSPs) include tumor-associated fibroblasts (TAFs) or cancer-associated fibroblasts (CAFs), extracellular matrix (ECM) proteins, T cells, tumor-associated macrophages (TAMs), myelosuppressor cells, and invasive immune and inflammatory cells such as blood and lymphatic vessels. These cells assist in the growth and development of tumors through mechanisms such as growth factor secretion, immunosuppression, metastasis, and resistance.

[0004] TAFs, or CAFs, are one of the major types of cells present in the tumor stroma and play several crucial roles in promoting tumor growth. These functions include ECM generation, remodeling, and cytokine secretion. These result in angiogenesis that promotes tumor growth, secretion of signaling factors that increase chemoresistance, a denser tumor stroma that provides physical blockage against immune cells, and increased cell motility that directs metastasis. In some cases, these processes resemble the behavior of pathogenic fibroblasts in fibrous diseases.

[0005] In some cases, a frequently observed marker of carcinoma-associated tumors (CAFs) is fibroblast-activating protein alpha (FAPα). FAPα is a serine protease found (primarily) on the cell surface of activated fibroblasts in affected cells and tissues such as fibrous diseases, inflammatory diseases, and / or cancers (e.g., fibrosis, rheumatoid arthritis, wound healing, and cancer). Over 90% of epithelial cancers show FAPα expression in immunohistochemical (IHC) staining. Further FAPα expression has been found in a subset of primary glioma cell cultures and TAMs. Recently, FAPα expression has been detected in at least 28 different types of human cancer. However, FAPα expression is very low or absent in most adult tissues. Therefore, because this expression is limited to the surface of affected cells such as carcinomas, FAPα has been specifically identified as a receptor for selective delivery of therapeutic agents to tumors through ligand targeting.

[0006] For the treatment of various cancers, fibrous disorders, and inflammatory diseases, radiation therapy, chemotherapy, and other therapies aimed at killing tumor cells can be considered. However, in many cases, such therapies are not used as first-line treatments due to the potential side effects (e.g., systemic) that may occur. Consequently, there is a need for targeted therapies, such as therapies using targeted radiotherapies and / or chemotherapy agents that may target affected cells and tissues, and can treat diseases (e.g., cancer, fibrous disorders, and / or inflammatory diseases) with minimal or reduced off-target or systemic effects. [Overview of the project]

[0007] Compounds of formula X (e.g., conjugates): A mx -L-B' (X) (In the formula, A is a radical (e.g., having a molecular weight of less than 10,000) of the fibroblast-activating protein alpha (FAPα) ligand (i.e., the target-directing portion), L is a linker that links one or more A groups to B' (for example, by a first covalent bond linking L to A and a second covalent bond linking L to B') (for example, difunctionalized or trifunctionalized), B' is a photodynamic agent, radioimaging agent, radiotherapy agent, chemotherapeutic agent, antifibrotic agent or anticancer agent (e.g., an anticancer agent effective against cancer cells or cancer-associated fibroblasts, myofibroblasts or other tumor microenvironment factors) (e.g., its radical), (mx is 1-6) It will be provided.

[0008] Similarly, compounds of formula I (e.g., conjugates): AL-B'(I) (In the formula, A contains an FAPα ligand (i.e., a target-directing moiety) (e.g., its radical), L includes a linker that links one or more A groups to B' (for example, a difunctionalized or trifunctionalized linker), B' includes photodynamic agents, radioimaging agents, radiotherapy agents, chemotherapeutic agents, antifibrotic agents, or anticancer agents (e.g., anticancer agents effective against cancer cells or cancer-associated fibroblasts, myofibroblasts, or other tumor microenvironment factors) (e.g., their radicals). It will be provided.

[0009] Compounds of formula I' (e.g., conjugates):

[0010] [ka] (In the formula, A is a radical of the FAPα ligand (i.e., the target-directing moiety) (for example, having a molecular weight of less than 10,000), L is a trivalent linker, B' is a radical of a phosphoinositide 3-kinase (PI3K) inhibitor, a chelate group optionally bound to an isotope, or a group covalently bound to an isotope, and the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is an albumin-binding ligand, polyethylene glycol. n (PEG) n (n is an integer between 0 and 32, a peptide, peptidoglycan, or sugar radical) It will be provided.

[0011] Similarly, compounds of formula (II) (e.g., conjugates):

[0012] [ka] (In the formula, A contains a radical of the FAPα ligand (i.e., a target-directing moiety) (for example, having a molecular weight of less than 10,000), L contains a trivalent linker, B’ contains a radical of a PI3K inhibitor, a chelating group optionally bonded to an isotope or a group covalently bonded to an isotope, and the isotope (or metal) is suitable for radiation imaging, radiation therapy or magnetic resonance imaging. C’ is an albumin-binding ligand, (PEG) n (n is an integer from 0 to 32), a peptide, a peptidoglycan or a sugar radical) is provided. Similarly, a compound represented by the structure of formula (V):

[0013]

Chemical formula

[0014] A compound represented by the structure of formula (X): A m -L-B’ (X) (wherein, A is a radical of a FAPα ligand of formula X-B:

[0015]

Chemical formula

[0016] In certain embodiments, such a compound further comprises C', where L is linked to one or more A groups and B', and C' is an albumin-binding ligand, (PEG) n (where n is an integer between 0 and 32), it is a peptide, peptidoglycan, or sugar radical.

[0017] In some embodiments, B' is a radical of a PI3K inhibitor, a chelate group optionally bound to an isotope, or a group covalently bound to an isotope, wherein the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0018] Compounds represented by the structure of formula (I'):

[0019] [ka] (In the formula, A is the FAP ligand of formula XB (i.e., the target-directing portion, e.g., and not limited to, the FAPα ligand):

[0020] [ka] It is a radical of, During the ceremony, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). J is C(R J ) 0~3 And R J Each of these is independently either H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6Each is independently selected from the group consisting of alkyl groups, R 5 , R 6 , R 7 and R 8 These are independently selected from the group consisting of H, alkyl, and halo. R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6 Each element is independently selected from the group consisting of alkyl, F, Cl, Br, and I. L is a trivalent linker, B' is a radical of a chelate group optionally bound to an isotope, and the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is an albumin-binding ligand, (PEG) n (n is an integer between 0 and 32, a peptide, peptidoglycan, or sugar radical) It is also provided.

[0021] In certain embodiments of the compound, A is

[0022] [ka] That's fine.

[0023] In certain embodiments of the compound, A is

[0024] [ka] That's fine.

[0025] In at least one embodiment of the compound, A is

[0026] [ka] Includes.

[0027] Alternatively, A is,

[0028] [ka] It can include, where X is between 1 and 20.

[0029] In a particular embodiment, A is

[0030] [ka] Includes.

[0031] B' may be a radical of a chelate group optionally bound to an isotope (or metal) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

[0032] B' is,

[0033] [ka] You can choose from these, each optionally bound to an isotope (or metal) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

[0034] In a particular embodiment, B' is

[0035] [ka] They are selected from among, each optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging.

[0036] B' may be a radical of a chelate group bonded to an isotope suitable for, for example, PET imaging, SPECT imaging, other radiographic imaging techniques, magnetic resonance imaging, or radiotherapy. In certain embodiments, B' may include a radical of DOTA. B' may include a radical of DOTA with a chelated (or metal-chelated) isotope. B' may include a radical of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid. B' may be a radical of a group covalently bonded to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging. B' is

[0037] [ka] It may include a base selected from the following.

[0038] B' is optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging.

[0039] [ka] It may contain a radical of a radioimaging isotope, radiotherapy isotope, or magnetic resonance isotope. In certain embodiments, B' includes a chelate group and a radioimaging isotope, radiotherapy isotope, or magnetic resonance isotope (or metal) bonded to the chelate group.

[0040] In a particular embodiment, B' comprises a radical of a radiation imaging isotope, a radiotherapy isotope or a magnetic resonance isotope, or a chelate group, and an isotope (e.g., a metal) bonded to the chelate group, where the isotope is 18 F, 32 P, 44 Sc, 47 Sc, 52 Mn, 55 Co,64 Cu, 67 Cu, 67 Ga, 68 Ga, 86 Y, 89 Sr, 89 Zr, 90 Y, 99m Tc, 111 In, 114m In, 117m Sn, 124 I, 125 I, 131 I, 149 Tb, 153 Sm, 152 Tb, 155 Tb, 161 Tb, 169 Er, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 224 Ra, 225 Ab, 225 Ac or 227 Th. The isotope of B' may be 111 In. The isotope of B' may be 177 Lu. In certain embodiments, the isotope is 11 C, 13 C, 13 N, 15 O, 60 Co and 123 I selected from the group consisting of.

[0041] A of the compound can have a binding affinity of about 1 nM to about 25 nM for FAPα. In certain embodiments, A has a binding affinity of about 1 nM to about 25 nM for FAP.

[0042] L of the compound can include a non-cleavable linker. L can include a cleavable linker. In certain embodiments, L is PEG nIt includes n=0~36. L may contain peptides. L may contain peptidoglycans. In certain embodiments, L is

[0043] [ka] Includes.

[0044] L is

[0045] [ka] It can include...

[0046] C' may be a radical of an albumin-binding ligand, with the following structure:

[0047] [ka] It holds.

[0048] In a particular embodiment, C' is a radical of an albumin-binding ligand, with the following structure:

[0049] [ka] It holds.

[0050] C' is, for example, the following:

[0051] [ka] It may be one of the radicals.

[0052] In some embodiments, C' may be a radical of an albumin-binding small protein scaffold, including ABD035, ABDCon, DARPin, dsFv CA645, nanobodies, and VNAR(E06). C' is PEG nIt may also be a peptidoglycan, where n is an integer between 0 and 32. C' may be a peptide. C' may be a peptidoglycan. C' may be a sugar.

[0053] In a particular embodiment, C' is

[0054] [ka] That is the case.

[0055] C' is,

[0056] [ka] That's fine.

[0057] In a particular embodiment, the compound has the structure of formula (V):

[0058] [ka] It can be represented by, During the ceremony, L is a linker containing at least one carbon atom, p is 0, 1, 2, or 3.

[0059] However, in some embodiments, the compound is as follows:

[0060] [ka] isn't it.

[0061] p may be 2. p may be 0.

[0062] Furthermore, L is

[0063] [ka] It can include, During the ceremony, m is an integer from 1 to 9. n is an integer between 1 and 32. q is an integer between 0 and 4. s is an integer between 0 and 4.

[0064] In a particular embodiment, the compound is

[0065] [ka] And, In the formula, t is either 0 or 1, and u is an integer between 2 and 12.

[0066] In some embodiments, the compound is

[0067] [ka] And, m is an integer between 1 and 4.

[0068] In some embodiments, the compound is given by formula:

[0069] [ka] That is the case.

[0070] In some embodiments, the compound is given by formula:

[0071] [ka] That is the case.

[0072] In some embodiments, the compound is given by formula:

[0073] [ka] That is the case.

[0074] In some embodiments, the compound is given by formula:

[0075] [ka] That is the case.

[0076] In some embodiments, the compound is given by formula:

[0077] [ka] That is the case.

[0078] In some embodiments, the compound has the following structure:

[0079] [ka] Each of these isotopes (or metals) is sometimes bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0080] In some embodiments, the compound has the following structure:

[0081] [ka] Each of these isotopes (or metals) is sometimes bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0082] In some embodiments, the compound has the following structure:

[0083] [ka] Each of these isotopes (or metals) is sometimes bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0084] In some embodiments, the compound has the following structure:

[0085] [ka] Each of these isotopes (or metals) is sometimes bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0086] In a particular embodiment, the compound has the following structure:

[0087] [ka] And, Each is sometimes bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0088] In certain embodiments, the compound may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, forming the following structure:

[0089] [ka] That is the case.

[0090] In certain embodiments, this compound may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, and may have the following structure:

[0091] [ka] It holds.

[0092] In certain embodiments, the compound may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, forming the following structure:

[0093] [ka] It can have.

[0094] In certain embodiments, the compound may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, forming the following structure:

[0095] [ka] It can have.

[0096] The compounds described herein have the following structure:

[0097] [ka] It may have, each optionally bound to an isotope (or metal) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

[0098] In certain embodiments, the compounds herein may be conjugated to an isotope (or metal) suitable for radioimaging, radiotherapy, or magnetic resonance imaging, and have the following structure:

[0099] [ka] It can have.

[0100] The compounds described herein may be conjugated to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, and may have the following structures:

[0101] [ka] It can have.

[0102] The compounds described herein may be conjugated to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, and may have the following structures:

[0103] [ka] It can have.

[0104] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0105] [ka] It can have.

[0106] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0107] [ka] It can have.

[0108] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0109] [ka] It can have.

[0110] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0111] [ka] It can have.

[0112] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0113] [ka] It can have.

[0114] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0115] [ka] It can have.

[0116] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0117] [ka] It can have.

[0118] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0119] [ka] It can have.

[0120] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0121] [ka] It can have.

[0122] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0123] [ka] It can have.

[0124] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0125] [ka] It can have.

[0126] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0127] [ka] It can have.

[0128] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0129] [ka] It can have.

[0130] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0131] [ka] It can have.

[0132] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0133] [ka] It can have.

[0134] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0135] [ka] It can have.

[0136] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0137] [ka] It can have.

[0138] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0139] [ka] It can have.

[0140] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0141] [ka] It can have.

[0142] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0143] [ka] It can have.

[0144] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0145] [ka] It can have.

[0146] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0147] [ka] It can have.

[0148] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0149] [ka] It can have.

[0150] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0151] [ka] It can have.

[0152] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0153] [ka] It can have.

[0154] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0155] [ka] It can have.

[0156] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0157] [ka] It can have.

[0158] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0159] [ka] It can have.

[0160] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0161] [ka] It can have.

[0162] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0163] [ka] It can have.

[0164] This compound has the following structure, optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging:

[0165] [ka] It can have.

[0166] Certain compounds described herein have the structure of formula (X): A m -L-B' (X) Represented by, During the ceremony, A is the structure of formula XX:

[0167] [ka] Represented by,

[0168] [ka] teeth,

[0169] [ka] It has a structure selected from the group consisting of, n = 1 to 5, n' = 1 to 5, Ring C is of arbitrary choice, X, Y, and Z in ring B are independently selected from O, N, and S, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, where at least one of X' and Y' is N. P is the bond site of ring C (if present) to L or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl. L is a linker if it exists. B' is a radical of an imaging agent or a radical of a therapeutic agent. m = 1 to 6.

[0170] This compound may further contain C', where L is linked to one or more A groups and B', and C' is an albumin-binding ligand, (PEG) n (where n is an integer between 0 and 32), it is a peptide, peptidoglycan, or sugar radical.

[0171] In certain embodiments, B' is a radical of a radioimaging agent, radiotherapy agent, or magnetic resonance imaging agent.

[0172] B' may be aromatic. In a particular embodiment, n=2, n'=1, and A is given by equation (XZ):

[0173] [ka] It is represented by [this].

[0174] In a particular embodiment, n = 0 to 4, and A has the structure of equation XY:

[0175] [ka] It is represented by [this].

[0176] In a particular embodiment, the compound has the following structure:

[0177] [ka] It holds.

[0178] In a particular embodiment, the compound has the following structure:

[0179] [ka] It holds.

[0180] In a particular embodiment, the compound has the following structure:

[0181] [ka] It holds.

[0182] In a particular embodiment, the compound has the following structure:

[0183] [ka] It holds.

[0184] The L of the compound may contain one or more linker groups, each independently selected from the group consisting of alkyl (alkylene), heteroalkyl (alkylene), heterocycloalkyl (alkylene), heteroaryl, aryl, alkoxy, thioether, disulfide, carboxylic acid, anhydride, carbonate, carbamate, thioether, sugar, peptide, and peptidoglycan. L may be a single linker. L may contain two or more linker groups. In certain embodiments, each linker group may be PEG.

[0185] In certain embodiments, L of the compound is (L 1 ) p -W-(L 2 ) q And L 1 This is the first linker, L 2 is the second linker, W is the third linker, and p=1~5 and q=1~5. Each L 1 and each L 2 It may independently contain one or more linker groups, each of which is independently selected from the group consisting of alkyl (alkylene), heteroalkyl (alkylene), heterocycloalkyl (alkylene), heteroaryl, aryl, alkoxy, thioether, disulfide, carboxylic acid, anhydride, carbonate, carbamate, thioether, sugar, peptide, and peptidoglycan. In certain embodiments, each L 1 and each L 2 W may independently contain one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), amide, phenyl, and triazole. In certain embodiments, W has an amine core, an aromatic core, or an alkylene core.

[0186] L, L of the compound 1 , L 2 or L 1 and L 2It can have a length of 5 angstroms to 200 angstroms. In some embodiments, L, L 1 , L 2 or L 1 and L 2 Structure:

[0187] [ka] It may include at least one linker group having a , where n is an integer from 0 to 10.

[0188] L, L 1 , L 2 or L 1 and L 2 Structure:

[0189] [ka] It may include at least one linker group having L, L 1 , L 2 or L 1 and L 2 Structure:

[0190] [ka] It may contain at least one linker group having , and n=1 to 32.

[0191] In certain embodiments (e.g., including triazole), B' is a radical of a dye. The dye may be, for example, fluorescent. B' may be a radical of an anticancer agent. B' may be a radical of an antifibrotic agent. B' may be a radical of a PI3K inhibitor. B' may be a radical of a radioimaging agent containing a chelated radioisotope. In certain embodiments, the radioisotope is 99m Tc, 111 In, 18 F, 68 Ga, 124 I,125 I and 131 Selected from the group consisting of I. In a particular embodiment, the radioactive isotope is 32 P, 89 Sr, 90 Y, 153 Sm, 169 Er, 177 Lu, 186 Re, 188 Re, 149 Tb, 211 At, 212 Bi, 213 Bi and 225 Selected from the group consisting of Ac. B' may be a radical of a dye. B' may be a radical of a fluorescent dye. B' may be a radical of an anticancer drug. B' may be a radical of an antifibrotic agent.

[0192] A ligand is also provided. In a particular embodiment, the ligand is for FAP and comprises an isoindoline scaffold into which the triazole moiety is introduced, and this ligand has a Schrödinger molecular docking score of at least about -8.0 kcal / mol.

[0193] In a particular embodiment, compound A has the structure X-X':

[0194] [ka] Represented by,

[0195] [ka] It has the formula XB, P is a bond site to L or B' in formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0196] In a particular embodiment, A has the structure of equation XY:

[0197] [ka] Represented by,

[0198] [ka] It has the formula XB, n = 0 to 4, X, Y, and Z are independently selected from O, N, and S, respectively, provided that at least one of X and Y is N, or Z is N. X' and Y' are independently selected from O, N, and S, respectively, provided that at least one of X' and Y' is N, or Z is N. P is a bond site to L or B' in formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0199] A is the structure of equation XZ:

[0200] [ka] It can be further expressed by,

[0201] [ka] It has the formula XB, X, Y, and Z are independently selected from O, N, and S, respectively, provided that at least one of X and Y is N, or Z is N. X' and Y' are independently selected from O, N, and S, respectively, provided that at least one of X' and Y' is N, or Z is N. P is a bond site to L or B' in formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0202] A pharmaceutical composition comprising any of the compounds described herein is provided. Such a pharmaceutical composition may, for example, include any one of the compounds described herein and a pharmaceutically acceptable carrier.

[0203] A method for imaging cancer or fibrosis (e.g., pulmonary fibrosis, renal fibrosis, or hepatic fibrosis) in subjects having cancer or fibrosis is further provided. The method comprises the step of administering an effective amount of a compound or a pharmaceutical composition containing the compound to a subject in need. The method may further comprise the step of imaging the subject. In certain embodiments, the method further comprises the step of generating an image of cancer or fibrosis in the subject.

[0204] A method for treating fibrosis in a subject is further provided. This method includes the step of administering an effective amount of a compound or a pharmaceutical composition containing the compound to a subject in need. The fibrosis can be selected from, for example, pulmonary fibrosis, renal fibrosis, and hepatic fibrosis.

[0205] Further methods for treating inflammatory diseases or disorders are provided. These methods include administering a therapeutically effective amount of a compound or a pharmaceutical composition containing the compound (for example, as described herein) to a subject in need.

[0206] Methods for treating cancer are also provided. In certain embodiments, a method for treating cancer includes the step of administering a therapeutically effective amount of the compound herein or a pharmaceutical composition containing said compound to a subject in need. The cancer can be selected from the group consisting of lung cancer, breast cancer, colorectal cancer, cervical cancer, and brain cancer (e.g., glioblastoma). In certain embodiments, the method further includes a step performed for chemotherapy or radiotherapy (or, for example, a step performed for additional treatment of cancer).

[0207] Furthermore, a method is provided for improving the affinity of a ligand to FAP, using a ligand that includes an isoindoline scaffold. In certain embodiments, the method includes the step of introducing a triazole moiety into the isoindoline scaffold of the ligand by molecular modeling to achieve a higher Schrödinger molecular docking score. [Brief explanation of the drawing]

[0208] [Figure 1-1] This document shows examples of both the demonstrated and predicted structures of various fibroblast-activating proteins (FAPs). Structures FAP-3000 to FAP-3017 are synthetic examples, while FAP-3018 to FAP-3029 are predicted structures. [Figure 1-2] Same as above. [Figure 1-3] Same as above. [Figure 1-4] Same as above. [Figure 1-5] Same as above. [Figure 1-6] Same as above. [Figure 1-7] Same as above. [Figure 1-8] Same as above. [Figure 1-9] Same as above. [Figure 1-10] Same as above. [Figure 2]Figure 2A illustrates the substitution of FAP ligands conjugated to rhodamine fluorescent dye derived from HEK-FAP cells with FAP ligands conjugated to DOTA without an albumin binder (FAP-3000) at a certain concentration range. Figure 2B illustrates the same substitution assay with FAP ligands conjugated to DOTA with an iodobenzenealbumin binder (FAP-3001). Figure 2C illustrates the same substitution assay with FAP ligands conjugated to NOTA with a fluorobenzenealbumin binder (FAP-3002). Figure 2D illustrates the same substitution assay with FAP ligands conjugated to NOTA with a chlorobenzenealbumin binder (FAP-3003). Figure 2E illustrates the same substitution assay with FAP ligands conjugated to DOTA with a shorter PEG(4) spacer (FAP-3015). Figure 2F illustrates the same substitution assay with a longer PEG(12) spacer conjugated with an FAP ligand (FAP-3016). Figure 2G illustrates the same substitution assay with a monofluoroanalog of the FAP ligand (FAP-3017). [Figure 3-1] Figure 3A shows the radiolabeling data for indium-111 in FAP-3000. [Figure 3-2] Figure 3B shows the radiolabeling data for lutetium-177 in FAP-3000. [Figure 3-3] Figure 3C shows the radiolabeling data for indium-111 in FAP-3001. [Figure 3-4] Figure 3D shows the radiolabeling data for lutetium-177 in FAP-3001. [Figure 4] Figure 4A illustrates the binding curve of 111In-FAP-3000 to cancer-associated fibroblast cell line Hs894 over a range of concentrations, in the absence or presence of 100× excess FAP ligand as competitive blockade. Figure 4B illustrates the same binding curve assay using 111In-FAP-3001. [Figure 5]Figure 5A illustrates the administration study of 111In-FAP-3000 using single-photon emission tomography / computed tomography (SPECT / CT) with the same amount of radioactivity but different numbers of molecules in nude mice with HT29 tumors. Figure 5B illustrates the administration study of 111In-FAP-3001 using SPECT / CT in nude mice with HT29 tumors with the same amount of radioactivity but different numbers of molecules in nude mice with HT29 tumors. [Figure 6] Figure 6A illustrates the administration study of 111In-FAP-3000 using single-photon emission computed tomography (SPECT / CT) with the same amount of radioactivity but different numbers of molecules in nude mice with 4T1 tumors. Figure 6B illustrates the administration study of 111In-FAP-3001 using SPECT / CT in nude mice with 4T1 tumors, with the same amount of radioactivity but different numbers of molecules in nude mice with 4T1 tumors. The maximum intensity of radioactivity is identified by the white circles in each image. [Figure 7] Figure 7A illustrates the retention study of 111In-FAP-3001 using SPECT / CT in BALB / c mice with 4T1 tumors. Figure 7B illustrates the retention study of 111In-FAP-3001 using SPECT / CT in mice with KB tumors. The highest intensity of radioactivity is identified by the white circles in each image. [Figure 8] Figure 8A illustrates the in vivo distribution of 177Lu-FAP-3000 at various time points in BALB / c mice with 4T1 tumors. Figure 8B illustrates the in vivo distribution of 111In-FAP-3001 at various time points in BALB / c mice with 4T1 tumors. Standard error bars are shown in both Figures 8A and 8B. [Figure 9]Figure 9A illustrates a radiotherapy study using two different doses of Lu-177-labeled 177In-FAP-3001 (0.25 mCi vs. 0.5 mCi) in BALB / c mice injected on day 0, using a 4T1 tumor growth chart. Figure 9B tracks the relative body weight of the mice. Figure 9C shows a SPECT / CT scan of one of the treated mice 24 hours after injection, with the highest intensity of radioactivity identified by a white circle in each image. Standard error bars are shown in both Figures 9A and 9B. [Figure 10] Figure 10A illustrates a radiotherapy study using 0.5 mCi radiolabeled 177In-FAP-3000, using a KB tumor growth chart, in nude mice injected on day 0. Figure 10B illustrates the survival curve for the same study. Figure 10C tracks the relative body weight of mice in the same study. Standard error bars are shown in both Figures 10A and 10C. [Figure 11] Figure 11A illustrates a radiotherapy study using 0.5 mCi radiolabeled 177In-FAP-3001, using a KB tumor growth chart, in nude mice injected on day 0. Figure 11B illustrates the survival curve for the same study. Figure 11C tracks the relative body weight of the mice in the same study. Standard error bars are shown in both Figures 11A and 11C. [Figure 12-1] Figure 12A illustrates a radiotherapy study using 0.25 mCi radiolabeled 177In-FAP-3001, with an HT29 tumor growth chart, in nude mice injected on day 0. Figure 12B illustrates the survival curve for the same study. Figure 12C tracks the relative body weight of the mice in the same study. [Figure 12-2] Figure 12D shows a photograph of the tumor dissected after euthanasia. The standard error bars are shown in both Figures 12A and 12C. [Figure 13]Figure 13A illustrates a radiotherapy study using two different doses of Lu-177-labeled 177In-FAP-3001 (0.25 mCi vs. 0.5 mCi), injected on day 0, with a U87MG tumor growth chart in nude mice. Figure 13B illustrates the survival curves for the same study. Figure 13C tracks the relative body weight of mice in the same study. Standard error bars are shown in both Figures 13A and 13C. [Figure 14] Images of tissue sections taken from various organs of interest in the control and treatment (0.5 mCi) groups, stained with H&E for evaluation, are shown. [Figure 15-1] Figure 15A illustrates the radiotherapy study using 177In-FAP-3001 (either 1.5 mCi injected on day 0, or two different doses, i.e., 1.5 mCi + 0.60 mCi injected on days 0 and 3, respectively). Figure 15B illustrates the survival curve for the same study. Figure 15C tracks the relative body weight of mice in the same study. [Figure 15-2] Figure 15D shows SPECT / CT scans of one of the treated mice at various time points after injection. The standard error bars are shown in both Figures 15A and 15C. [Figure 16] Figure 16A illustrates a radiotherapy study with 1.5 mCi radiolabeled 177In-FAP-3001, injected on day 0 and then measured by two researchers in a blinded manner. Figure 16B illustrates the survival curve for the same study. Figure 16C tracks the relative body weight of mice in the same study. Standard error bars are shown in both Figures 16A and 16C. [Figure 17] Figure 17A illustrates a radiotherapy study using 1.5 mCi radiolabeled 177In-FAP-3001 versus 177In-FAP-3005 injected on day 0. Figure 17B illustrates the survival curve for the same study. Figure 17C tracks the relative body weight of mice in the same study. Standard error bars are shown in both Figures 17A and 17C. [Figure 18]Figure 18A shows a summary of the histopathology of autopsy tissues derived from radiotherapy treatment with 177Lu-FAP-3001 at different doses and various time points after injection. Figure 18B shows images of tissue sections taken from various organs of interest in the control and treatment (1.5 mCi) groups at various time points after injection, stained with H&E for evaluation. [Figure 19] SPECT / CT scans of a pulmonary fibrosis model administered 111In-FAP-3001 24 hours after injection are shown. The upper arrows indicate the lungs, and the lower arrows indicate the kidneys. [Figure 20-1] Figures 20A and 20B illustrate the structures of various fibroblast-activating protein (FAP)-phosphoinositide 3-kinase (PI3K) (FAP5-PI3K) inhibitors. [Figure 20-2] Same as above. [Figure 21] The results for FAP5-PI3K inhibitors after 24 hours are shown. [Figure 22] The results for FAP5-PI3K inhibitors after 48 hours are shown. [Figure 23] Figures 23A and 23B show graphical data related to the inhibition of Akt phosphorylation by the FAP5-PI3K inhibitor 24 hours after incubation (Figure 5A) and 48 hours after incubation (Figure 5B). [Figure 24] The mass spectrometry data confirming the formation of the desired compound are shown in the diagram. [Figure 25] The mass spectrometry data confirming the formation of the desired compound are shown in the diagram. [Figure 26] Molecular models of a known FAP inhibitor (FAP inhibitor 1) and FAP-4001 are illustrated, illustrating that docking them increases the interaction between FAP and FAP-4001. [Figure 27-1] Molecular models of various compounds and their interactions with FAP when docked are illustrated. [Figure 27-2] Same as above. [Figure 27-3] Same as above. [Figure 27-4] Same as above. [Figure 27-5] Same as above. [Figure 28-1] Exemplary structures of various FAP triazole scaffolds are shown. [Figure 28-2] Same as above. [Figure 29] The following shows exemplary structures of various compounds that do not contain a linker. [Figure 30] The following shows exemplary structures of various compounds, including PEG linkers. [Figure 31] This shows exemplary structures of various compounds, including alkyl linkers. [Figure 32] Figures 32A and 32B illustrate fluorescence images (panels I and iv) and white light images (panels ii and v) integrated into panels ii and vi, taken from studies internalizing FAP-4004 (Figure 32A) and FAP-4003 (Figure 32B) into HT1080-FAP cells. Figures 32C and 32D show graphical data regarding the binding affinity and specificity of FAP-4004 (Figure 32C) and FAP-4002 (Figure 32D) as described herein. [Figure 33] Figures 33A-33C illustrate graphical data related to the analysis of the ability of the FAP-targeting ligand FAP-4002 to inhibit the closely related dipeptidyl peptidases FAP (Figure 33A), PREP (Figure 33B), and DPP-IV (Figure 33C). [Figure 34-1] Figures 34A to 34C illustrate data confirming the formation of certain compounds described herein. Figures 34A and 34B show ¹H nuclear magnetic resonance (¹H NMR) spectroscopy data for compound 16, and Figure 34C shows ¹H NMR (D²O) spectral data for FAP-4003. [Figure 34-2] Same as above. [Figure 34-3] Same as above. [Figure 35] The excitation (Ex) and emission (Em) spectra of a 1 μM solution of FAP-4003 in PBS, pH 7.4 are shown. [Modes for carrying out the invention]

[0209] This disclosure relates to the preparation and use of compounds and compositions for reducing the tendency for off-target toxicity after administration of therapeutic and / or imaging agents (e.g., radioimaging agents, radiotherapeutic agents, chemotherapeutic agents, antifibrotic agents, or anticancer agents). The term “off-target toxicity” means damage to an organ or tissue, or weight loss of the subject that is undesirable to the treating physician, or any other effect on the subject that is a potentially harmful indicator to the treating physician (e.g., B-cell aplasia, fever, hypotension, or pulmonary edema). The terms “to treat,” “to be treated,” or “treated” (related to a disease or condition) are used in relation to methods for obtaining beneficial or desired outcomes, such as clinical outcomes. Such outcomes include, but are not limited to, one or more of the following: improvement of a disease-related condition, cure of a disease, reduction of disease severity, delay of disease progression, reduction of one or more symptoms related to a disease, improvement of the quality of life of a person with a disease, extension of survival, and / or preventive (e.g., inhibitory) treatments. In reference to cancer, the terms “to treat,” “to treat,” “to be treated,” or “treatment” may further mean reduction in tumor size, complete or partial removal of the tumor (e.g., complete or partial response), stabilization of the disease, prevention of cancer progression (e.g., progression-free survival), or any other effect on cancer that a physician considers to be a therapeutic or preventive treatment of cancer.

[0210] The compounds herein may comprise a fibroblast-activating protein (FAP) targeting ligand (or its radical) bound to a linker comprising one or more linker groups (e.g., three linker groups), the linker being further bound to a therapeutic or imaging agent. FAP is a type II membrane-bound serine protease that cleaves proline-amino acid peptide bonds and can be expressed in cancer-associated fibroblasts (CAFs) and myofibroblasts that produce collagen. In certain embodiments, the compounds can target therapeutic compounds to FAP expressing cancer, fibrous disease, or inflammatory disease. In certain embodiments, such improvements to the FAP targeting ligand scaffold can be further used in conjunction with an albumin-binding moiety to achieve targeted delivery of radiolabeled functional groups and other functional groups. In certain embodiments, the FAP targeting ligand is a high-affinity FAP ligand comprising a triazole moiety (or a derivative thereof) introduced into the ligand scaffold. In certain embodiments, the FAP target ligand is a high-affinity FAP ligand containing a triazole moiety (or a derivative thereof) and a phenyl ring, introduced into a ligand scaffold (e.g., an isoindoline ring scaffold). Unless otherwise specified, "high affinity" or "even higher affinity," relating to the ligand's affinity for the target, means a ligand having a Schrödinger molecular docking score of at least approximately -8.0 kcal / mol. In certain embodiments, the high-affinity FAP ligand exhibits improved affinity for FAP compared to a ligand that does not contain the introduced triazole moiety.

[0211] The compounds, compositions, and methods described herein will be described in more detail. To facilitate understanding of the principles presented herein, the embodiments illustrated in the drawings will be referenced, and specific terminology will be used to describe them. Nevertheless, it will be understood that the description of these embodiments is not intended to limit the scope. In contrast, this disclosure is intended to include alternatives, variations, and equivalents that may fall within the spirit and scope of this application as defined by the appended claims. As stated herein, the art may be illustrated and described in one or more preferred embodiments, but the compositions, compounds, and methods described herein may include a number of different configurations, forms, substances, and auxiliary materials.

[0212] compound Compounds (i.e., conjugates) of formula X: A mx -L-B' (X) (In the formula, A is a radical (e.g., having a molecular weight of less than 10,000) of the fibroblast-activating protein alpha (FAPα) ligand (i.e., the target-directing portion), L is a linker that links one or more A groups to B' (for example, by a first covalent bond linking L to A and a second covalent bond linking L to B') (for example, difunctionalized or trifunctionalized), B' is a photodynamic agent, radioimaging agent, radiotherapy agent, chemotherapeutic agent, antifibrotic agent or anticancer agent (e.g., an anticancer agent effective against cancer cells or cancer-associated fibroblasts, myofibroblasts or other tumor microenvironment factors) (e.g., its radical), (mx is 1-6) It will be provided.

[0213] In a particular embodiment, mx is 1. In a particular embodiment, mx is 2. In a particular embodiment, mx is 3. In a particular embodiment, mx is 4. In a particular embodiment, mx is 5. In a particular embodiment, mx is 6. mx may be 1-3, 2-4, or 1-5.

[0214] This disclosure also relates to compounds of formula I (i.e., conjugates): AL-B'(I) (In the formula, A contains an FAPα ligand (i.e., a target-directing moiety) (e.g., its radical), L includes a linker that links one or more A groups to B' (for example, a difunctionalized or trifunctionalized linker), B' includes photodynamic agents, radioimaging agents, radiotherapy agents, chemotherapeutic agents, antifibrotic agents, or anticancer agents (e.g., anticancer agents effective against cancer cells or cancer-associated fibroblasts, myofibroblasts, or other tumor microenvironment factors) (e.g., their radicals). Regarding.

[0215] Similarly, compounds of formula I (e.g., conjugates): AL-B' (I) (In the formula, A contains an FAPα ligand (i.e., a target-directing moiety) (e.g., its radical), L includes a linker that links one or more A groups to B' (for example, a difunctionalized or trifunctionalized linker), B' contains radiolabeled functional groups (e.g., their radicals) suitable for PET imaging, SPECT imaging, other radiographic imaging techniques, or radiotherapy. It will be provided.

[0216] Similarly, compounds represented by the structure of formula (X): AL-B'(X) (In the formula, A is given by equation XB:

[0217] [ka] It is the radical of the FAPα ligand, During the ceremony, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). J is C(R J ) 0~3 And R J Each of these is independently either H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 These are independently selected from the group consisting of H, alkyl, and halo. R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6 Independently selected from the group consisting of alkyl, F, Cl, Br, and I, L is a linker that connects A to B', B' is a therapeutic agent, radioimaging agent, radiotherapy agent, chemotherapeutic agent, antifibrotic agent, or anticancer agent. It will be provided.

[0218] In some embodiments, A in formula X has the structure of formula X-X':

[0219] [ka] Represented by,

[0220] [ka] It has the formula XB, P is a bond site to L or B' in formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0221] In some embodiments, A in formula X has the structure of formula XX:

[0222] [ka] Represented by,

[0223] [ka] It has the formula XB, n = 1 to 5, n' = 1 to 5, Ring C is of arbitrary choice, X, Y, and Z in ring B are independently selected from O, N, and S, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, where at least one of X' and Y' is N. P is the bond site of ring C (if present) to L or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0224] In some embodiments,

[0225] [ka] teeth,

[0226] [ka] It has a structure selected from the group consisting of the following.

[0227] In some embodiments, the compound further comprises C', L has C' linked to one or more A groups and B', C' is an albumin-binding ligand, polyethylene glycol. n (PEG) n (where n is an integer between 0 and 32), it is a peptide, peptidoglycan, or sugar radical.

[0228] Similarly, compounds represented by the structure of formula (I'):

[0229] [ka] (In the formula, A is given by equation XB:

[0230] [ka] It is the radical of the FAPα ligand (target-directing portion), During the ceremony, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). J is C(R J ) 0~3 And R J Each of these is independently either H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6 Each element is independently selected from the group consisting of alkyl, F, Cl, Br, and I. L is a trivalent linker, B' is a radical of a chelate group optionally bound to an isotope, and the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is an albumin-binding ligand, (PEG) n(n is an integer between 0 and 32, a peptide, peptidoglycan, or sugar radical) It will be provided.

[0231] In some embodiments, B' is a radical of a therapeutic or imaging agent, such as a phosphoinositide 3-kinase (PI3K) inhibitor, a chelating group optionally bound to an isotope (or metal), or a group covalently bound to an isotope (or metal), wherein the isotope (or metal) is suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

[0232] In some embodiments, B' is a radical of a chelate group optionally bonded to a metal, or a group covalently bonded to an isotope, wherein the metal or isotope is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0233] Compounds of formula I' (e.g., conjugates):

[0234] [ka] (In the formula, A is a radical of the FAPα ligand (i.e., the target-directing moiety) (for example, having a molecular weight of less than 10,000), L is a trivalent linker, B' is a radical of a therapeutic or imaging agent, such as a PI3K inhibitor, a chelate group optionally bound to an isotope (or metal), or a group covalently bonded to an isotope (or metal), wherein the metal or isotope (or metal) is suitable for radiation imaging, radiotherapy, or magnetic resonance imaging. C' is an albumin-binding ligand, (PEG) n (where n is an integer between 0 and 32, it is a peptide, peptidoglycan, or sugar radical.) It is also offered.

[0235] This disclosure also relates to compounds of formula (II) (e.g., conjugates):

[0236] [ka] (In the formula, A contains a radical of the FAPα ligand (i.e., a target-directing moiety) (for example, having a molecular weight of less than 10,000), L contains a trivalent linker, B' comprises a PI3K inhibitor, a chelate group optionally bonded to a metal, or a radical of a group covalently bonded to an isotope, wherein the metal or isotope is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is an albumin-binding ligand, (PEG) n (n is an integer between 0 and 32, including peptides, peptidoglycans, or sugar radicals) Regarding.

[0237] This disclosure also relates to compounds (i.e., conjugates) of formula (II):

[0238] [ka] (In the formula, A contains a radical of the FAPα ligand (i.e., a target-directing moiety) (for example, having a molecular weight of less than 10,000), L contains a trivalent linker, B' comprises a PI3K inhibitor, a chelate group optionally bound to an isotope (or metal), or a radical of a group covalently bonded to an isotope (or metal), wherein the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' contains the radical of an albumin-binding ligand. Regarding.

[0239] In some embodiments, the compound has the structure of formula (I'):

[0240] [ka] (In the formula, A is a radical of the FAPα ligand (i.e., the target-directing moiety) (for example, having a molecular weight of less than 10,000),

[0241] [ka] Includes, L is a trivalent linker, B' is a radical of a PI3K inhibitor, a chelate group optionally bound to an isotope (or metal), or a group covalently bonded to an isotope (or metal), wherein the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is an albumin-binding ligand, (PEG) n (n is an integer between 0 and 32, a peptide, peptidoglycan, or sugar radical) It is represented by [this].

[0242] In some embodiments, the compound has the structure of formula (I'):

[0243] [ka] (In the formula, A is a radical of the FAPα ligand (i.e., the target-directing moiety) (for example, having a molecular weight of less than 10,000),

[0244] [ka] Includes, L is a trivalent linker, B' is a radical of a chelate group optionally bonded to an isotope (or metal), and the said isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. C' is the radical of an albumin-binding ligand. It is represented by [this].

[0245] In some embodiments, the compound has the structure of formula (X): A m -L-B' (X) (In the formula, A is the structure of formula XX:

[0246] [ka] Represented by,

[0247] [ka] teeth,

[0248] [ka] A structure selected from the group consisting of, and any other FAP ligand structure described herein, n = 1 to 5, n' = 1 to 5, Ring C is of arbitrary choice, X, Y, and Z in ring B are independently selected from O, N, and S, respectively, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, respectively, where at least one of X' and Y' is N. P is the bond site of ring C (if present) to L or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl. L is a linker if it exists. B' is a radical of an imaging agent or a radical of a therapeutic agent. (m = 1 to 6) It is represented by [this].

[0249] This compound contains one or more chiral centers and can therefore give rise to enantiomers, diastereomers, and other stereoisomers defined as (R)- or (S)- with respect to absolute stereochemistry. Unless otherwise specified, all stereoisomers of this compound are intended to be considered. If this compound contains an alkene double bond, and unless otherwise specified, this is intended to include both E and Z geometric isomers (e.g., cis or trans). Similarly, all possible isomers, as well as their racemic and optically pure forms, and all tautomers are also intended to be included. The term “geometric isomer” refers to the E or Z geometric isomers (e.g., cis or trans) of an 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. Those skilled in the art will further recognize that this compound is “deuterated,” meaning that one or more hydrogen atoms can be replaced by deuterium.

[0250] The compound may be a monovalent conjugate (e.g., a compound comprising one binding ligand (as described elsewhere herein, e.g., one FAP-binding ligand)). The compound may be a divalent conjugate (e.g., a compound comprising one or more binding ligands (as described elsewhere herein, e.g., one or more FAP-binding ligands) conjugated to a therapeutic or imaging agent (e.g., via a linker) (as described elsewhere herein). The compound may be a polyvalent conjugate (e.g., a compound comprising two or more binding ligands (as described elsewhere herein, e.g., two or more FAP-binding ligands) conjugated to a multipoint linker).

[0251] Binding ligand / target-directing moiety (A) In certain embodiments, A in formula (X), (I), (I'), or (II) is a target-directing moiety / binding ligand. The binding ligand (also referred herein as a target ligand or target-directing moiety) may be a compound (or its radical) that binds to a biomolecule (e.g., polypeptide (e.g., enzyme)) localized to a particular cell, tissue, organ, etc. The binding ligand may be an FAP ligand (or its radical). The binding ligand may be a fibroblast-activating protein alpha (FAPα) ligand (or its radical). As used herein, “ligand” means a molecule, ion, or atom bound to a central atom or ion (e.g., a drug) of a compound. “Ligand” also includes binders that are neither agonists nor antagonists, and that do not possess either agonist or antagonist properties.

[0252] In certain embodiments, the target-directed moiety binds to activated fibroblasts expressing FAPα, such activated fibroblasts involved in cancer or inflammatory diseases.

[0253] The target-directed portion may be a radical of an FAPα ligand having a molecular weight of, for example, less than approximately 10,000, less than 7,500, less than 5,000, less than 2,500, less than 1,000, less than 760, less than 500; approximately 500 to approximately 10,000 g / mol, approximately 1,000 to approximately 7,500 g / mol, approximately 750 g / mol to approximately 1,500 g / mol, approximately 1,000 to approximately 5,000 g / mol, or approximately 500 to approximately 2,500 g / mol.

[0254] The target-directing portion can bind to activated fibroblasts that express FAPα, and such activated fibroblasts are involved in cancer or inflammatory diseases.

[0255] In a particular embodiment, A is a structure represented by formula (I-A1) or (I-A2):

[0256] [ka] It has, During the ceremony,

[0257] [ka] This is a functionalized 5-10 member nitrogen-containing aromatic or non-aromatic, monocyclic or bicyclic heterocycle, wherein the heterocycle optionally further contains 1-3 heteroatoms selected from oxygen, nitrogen, and sulfur. In formula (I-A1), Z is a bond, a substituted or unsubstituted alkylene (e.g., -CH2-), a substituted or unsubstituted amino (e.g., -NH-), -O-, or -S-. T is a substituted or unsubstituted methylene group (-CH2-), a substituted or unsubstituted amino group (-NH-), -O-, or -S-. R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, Q in equation (I-A2) 1 It is selected from the group consisting of -H, -CH3, -CH2OH, and -CH(CH3)2.

[0258] [ka] This is the binding site of the FAPα-binding ligand (e.g., via the linker L, or the imaging agent / therapeutic portion B'), and this binding site is a carbon atom of a 5-10 member nitrogen-containing aromatic or non-aromatic, monocyclic or bicyclic heterocyclic ring, 1 o Or 2 o This can be mediated by an amine, or a functionalized alkyl or cycloalkyl motif, as well as a stereoisomer or a pharmaceutically acceptable salt thereof.

[0259] A is a structure represented by formula IB:

[0260] [ka] It can have, During the ceremony, T is a substituted or unsubstituted methylene group (-CH2-), a substituted or unsubstituted amino group (-NH-), -O-, or -S-. R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -OC 1~6 Alkyl, -SC 1~6It is independently selected from the group consisting of alkyl, F, Cl, Br, and I.

[0261] In a particular embodiment, A is a structure represented by formula IC:

[0262] [ka] It has, During the ceremony, T is a substituted or unsubstituted methylene group (-CH2-), a substituted or unsubstituted amino group (-NH-), -O-, or -S-. R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -OC 1~6 Alkyl, -SC 1~6 It is independently selected from the group consisting of alkyl, F, Cl, Br, and I.

[0263] A is a structure represented by the following formula:

[0264] [ka] It can have, In the formula, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S-. R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -OC 1~6 Alkyl, -SC 1~6 Each element is independently selected from the group consisting of alkyl, F, Cl, Br, and I.

[0265] In a particular embodiment, A is a structure represented by formula XA:

[0266] [ka] It has, During the ceremony, Q is an aryl, heteroaryl, or heterocyclyl (e.g., including aryl ring and nonaryl ring structures) (e.g., a 5-10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle may optionally further include 1-3 heteroatoms selected from O, N, and S), Z is a bonded, substituted or unsubstituted C1-C3 alkylene (e.g., -CH2-), a substituted or unsubstituted heteroalkyl (e.g., an atom of 1-3 length), an amino (e.g., NH), -O-, or -S-. T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 The element is independently selected from the group consisting of H, alkyl, and halo.

[0267] In certain embodiments of formula (XA), Q is bonded to L (e.g., L or L1 (see the linker section below)). Q may be an aryl, heteroaryl, or heterocyclyl. Heterocyclyls may include aryl and nonaryl ring structures. Q may bond to L at the heteroalkyl, alkyl, or aryl position of Q. Q may bond to L at the aryl position of Q. Q may bond to L via a nitrogen atom (e.g., of L). Q may bond to L via a triazolyl or amide (e.g., of L). Heteroaryls may include aryl and nonaryl ring structures. Heteroaryls or heterocyclyls may contain 1 to 3 heteroatoms selected from O, N, and S. Heterocyclyls may contain 1 to 3 heteroatoms selected from O, N, and S. Q may be a 5 to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle (e.g., optionally including aryl and nonaryl ring structures). Q may be an N-linked heterocyclyl (e.g., optionally including aryl ring and non-aryl ring structures). Q may also be a C6-C9 N-linked heterocyclyl (e.g., optionally including aryl ring and non-aryl ring structures). The N-linked heterocyclyl is linked to Z via an N-heterocycloalkyl group. Q may also be an N-linked isoindolinyl (e.g., N is linked to Z in formula (XA)).

[0268] As stated above, in certain embodiments of formula (XA), Z is a bonded, substituted or unsubstituted C1-C3 alkylene, a substituted or unsubstituted heteroalkylene (e.g., an atom of length 1-3), an amino (e.g., NH), -O-, or -S-. Z may be bonded. Z may be a substituted methylene. Z may be -CH2-. Z may be a substituted ethylene. Z may be an oxo-substituted ethylene. Z may be -C(CO)CH2-. Z may be -CH2CH2-. Z may be a C1-C3 heteroalkylene.

[0269] In a particular embodiment, A is a structure represented by formula XB:

[0270] [ka] is or includes the same, During the ceremony, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). J is C(R J ) 0~3 And R J Each of these is independently either H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6It is independently selected from the group consisting of alkyl, F, Cl, Br, and I.

[0271] In a particular embodiment, A is a structure represented by formula XC:

[0272] [ka] is or includes the same, During the ceremony, T is a substituted or unsubstituted methylene (-CH2-), a substituted or unsubstituted amino (-NH-), -O-, or -S- (for example, the substituent of T is C1-C3 alkyl, haloalkyl, or halo). J is C(R J ) 0~3 And R J These are independently H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 These are independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 , R 6 , R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 , R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC1~6 Alkyl, -SC 1~6 It is independently selected from the group consisting of alkyl, F, Cl, Br, and I.

[0273] In some embodiments, J is C(R J ) 1~3 That is the case.

[0274] In formulas (XB), (XC), or (XD), J may bond to L of the compound (e.g., L or L1 (see the linker section below)). J may bond to L via a nitrogen atom. J may bond to L via a triazolyl or amide (e.g., L). J may bond to C(R J ) may be 2, R J Each of these is independently either H or alkyl, or R J Both combine to form an oxo. J may be a C1-C3 alkyl group. J may be a -CH2- group. J may be a -CH2CH2- group. J may be a C=O group. J may be a bond.

[0275] In formula (XB) or (XC), T may be a substituted or unsubstituted methylene group (e.g., -CH2-), a substituted or unsubstituted amino group (e.g., -NH-), -O-, or -S-. The substituent of T may be a C1-C3 alkyl group, a C1-C3 haloalkyl group, or (in the case of methylene) a halo group. T may be (-CH2-). The substituent of T may be a C1-C3 alkyl group, a haloalkyl group, or a halo group. T may be unsubstituted.

[0276] In certain embodiments of formula (XB) or (XC), R 1 and R 2 Each of these is independently selected from the group consisting of -H, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, -CONH2, and 5-tetrazolyl. 1 and R2 Each of these can be independently selected from the group consisting of H, -CN, -CHO, and -B(OH)2. 1 and R 2 Each of these can be independently selected from the group consisting of H, -CN, -CHO, and -CONH2. 1 H may also be used. 2 R may be -CN, -CHO, -B(OH)2, or -CONH2. 1 It may be H, and R 2 R may be -CN, -CHO, -B(OH)2, or -CONH2. 1 It may be H, and R 2 -CN may also be used. 1 It may be H, and R 2 -CHO is also acceptable. 1 It may be H, and R 2 R may also be -B(OH)2. 1 It may be H, and R 2 -CONH2 is also acceptable.

[0277] R of equation (XB) or (XC) 3 and R 4 These are -H, -OH, F, Cl, Br, I, and -C, respectively. 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 It can be independently selected from the group consisting of alkyl groups. 3 and R 4 Each of these can be independently -H or -F. 3 It may be H, and R 4 -F is also acceptable. 3 It may also be F, R 4 -F is also acceptable.

[0278] In a particular embodiment, R 1 H is R 2 is -CN, R 3 H is R 4is -F. In a particular embodiment, R 1 H is R 2 is -CN, R 3 F is R 4 is -F. Furthermore, R 1 H may also be R 2 It may also be -CHO, R 3 H may also be R 4 It can also be -F. 1 H may also be R 2 It may also be -CHO, R 3 It may also be F, R 4 It can also be -F. 1 H may also be R 2 R may also be -B(OH)2, 3 H may also be R 4 It can also be -F. 1 H may also be R 2 R may also be -B(OH)2, 3 It may also be F, R 4 It can also be -F. 1 H may also be R 2 H may be -CONH2, R3 may be H, R 4 It can also be -F. 1 H may also be R 2 It may also be -CONH2, R 3 It may also be F, R 4 -F is also acceptable.

[0279] R 5 , R 6 , R 7 and R 8 Each of these can be independently selected from the group consisting of H, alkyl, and halo. 5 , R 6 , R 7 and R 8 These can each be H.

[0280] R 9 , R 10 and R11 These are H and -C respectively. 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6 It can be independently selected from the group consisting of alkyl, F, Cl, Br, and I. 9 , R 10 and R 11 These are H and -C respectively. 1~6 R can be independently selected from the group consisting of haloalkyl, F, and Cl. 9 and R 11 H may also be R 10 H, -C 1~6 R may be a haloalkyl, F, or Cl. 9 and R 11 It may be H, and R 10 R may be H, -CF3, F, or Cl. 9 and R 11 It may be H, and R 10 -CF3 may also be used. 9 and R 11 It may be H, and R 10 It may also be F. 9 and R 11 It may be H, and R 10 It may be Cl. 9 , R 10 and R 11 H may also be used.

[0281] In a particular embodiment, A is a structure represented by formula (XD):

[0282] [ka] is or includes the same, During the ceremony, J is C(R J 2) 0~3 And R J Each of the 2s is either H or two or more R J 2 combine to form an oxo, R 1 It is selected from the group consisting of -CN, -CHO, and -B(OH)2, R 3 and R 4 These are independently selected from the groups consisting of -H and F, R 10 The element is selected from the group consisting of H, -CF3, F, Cl, Br, and I.

[0283] A may be bonded to L of the compounds herein via a nitrogen atom (for example, of L). A may be bonded to L via a triazolyl or amide (for example, of L).

[0284] In a particular embodiment, A is

[0285] [ka] That is the case.

[0286] A is

[0287] [ka] You can choose from the group consisting of these.

[0288] A in this compound also,

[0289] [ka] You can choose from the group consisting of these.

[0290] A is

[0291] [ka] That's fine.

[0292] A is

[0293] [ka] That's fine.

[0294] In some embodiments, A is

[0295] [ka] It is either or includes this.

[0296] In some embodiments, A is

[0297] [ka] It is either or includes this.

[0298] In some embodiments, A is

[0299] [ka] is or includes the same, X is between 1 and 20.

[0300] In some embodiments, A is

[0301] [ka] It is either or includes this.

[0302] In certain embodiments, compound A of the herein (e.g., a FAPα-binding ligand) may have a binding affinity to FAP (e.g., FAPα) in the range of about 1 nM to about 25 nM, such as 1 nM to about 25 nM or about 1 nM to about 25 nM.

[0303] In certain embodiments, A of the compound comprises a high-affinity FAP-binding ligand. Such a high-affinity FAP ligand may, in certain embodiments, include a triazole moiety or one or more derivatives thereof. In certain embodiments, A is an FAP ligand comprising a triazole moiety within an isoindoline scaffold. In certain embodiments, A is an FAP ligand comprising a triazole moiety and a phenyl ring. In certain embodiments, A is an FAP ligand comprising an ethyldiaminoaryltriazole moiety. In certain embodiments, A is an FAP ligand comprising an ethyldiaminoaryltriazole moiety and a phenyl ring. In certain embodiments, the triazole moiety may further comprise a primary or secondary amine, or a functionalized alkyl or cycloalkyl motif.

[0304] If the FAP ligand contains a triazole moiety (or a derivative thereof) (e.g., introduced into an isoindoline scaffold), further interaction with the FAP target may be achieved, resulting in a higher docking score in Schrödinger molecular docking calculations (see, for example, Figure 26) (compared to FAP ligands that do not have a triazole moiety and / or phenyl ring bound to the FAP ligand).

[0305] In a particular embodiment, a ligand for FAP is provided, comprising an isoindoline scaffold into which a triazole moiety is introduced, and having a Schrödinger molecular docking score of at least about -8.2 kcal / mol.

[0306] In a particular embodiment, A is

[0307] [ka] And, n = 1 to 5, n' = 1 to 5, Ring C is of arbitrary choice, X, Y, and Z in ring B are independently selected from O, N, and S, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, where at least one of X' and Y' is N. P is a bond site of ring C (if present) to the linker (L) or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0308] [ka] This may be any FAP ligand structure provided herein. In a particular embodiment,

[0309] [ka] teeth,

[0310] [ka] It is a structure selected from the group consisting of the following.

[0311] In certain embodiments, the B and C rings may each be functionalized 5-10 member nitrogen-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, the heterocycles may optionally further contain 1-3 heteroatoms selected from O, N, and S. In certain embodiments, if ring C is absent, the bond site of formula (X) to L or B' may be via any carbon atom of the 5-10 member nitrogen-containing B ring. Furthermore, the bond site of formula (X) to L or B' (e.g., P) may be between a functionalized alkyl or cycloalkyl motif of the B or C ring, or a primary or secondary amine.

[0312] In certain embodiments, the high-affinity FAP-binding ligand further comprises C'. C' may be linked to one or more A groups and B' by L. C' is, for example, an albumin-binding ligand, (PEG) n (where n is an integer between 0 and 32), may be a peptide, peptidoglycan, or sugar radical.

[0313] In certain embodiments, ring B is non-aromatic. In certain embodiments, ring B is aromatic.

[0314] In a particular embodiment, A is given by equation (XZ):

[0315] [ka] It is expressed by, in the formula, Ring C is of arbitrary choice, X, Y, and Z in ring B are independently selected from O, N, and S, respectively, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, respectively, where at least one of X' and Y' is N. P is a bond site of ring C (if present) to the linker (L) or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0316] In a particular embodiment, A is given by equation XY:

[0317] [ka] It is expressed by, in the formula, Ring C is of arbitrary choice, n = 0 to 4, X, Y, and Z in ring B are independently selected from O, N, and S, provided that at least one of X and Y is N, or Z is N. X' and Y' in ring C are independently selected from O, N, and S, where at least one of X' and Y' is N. P is a bond site of ring C (if present) to the linker (L) or B' of formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0318] In some embodiments, A has the structure of formula XZ:

[0319] [ka] Represented by,

[0320] [ka] It has formula XB (or any other FAP ligand structure described herein), X, Y, and Z are independently selected from O, N, and S, provided that at least one of X and Y is N, or Z is N. X' and Y' are chosen independently from O, N, and S, provided that at least one of X' and Y' is N, or Z is N. P is a bond site to L or B' in formula (X), and is selected from the group consisting of -H, -OH, -NH2, -COOH, -CONH2, -CHO, -N3, -CN, -B(OH)2, -C(O)alkyl, -C(O)aryl, -C=CC(O)aryl, and -C=CS(O)2aryl.

[0321] In some embodiments, compound A of the herein (e.g., a high-affinity FAP ligand) may have a Schrödinger molecular docking score of at least about -8.2 kcal / mol. In certain embodiments, compound A of the herein may have a Schrödinger molecular docking score of at least about -11.5 kcal / mol.

[0322] Therapeutic agents and / or imaging agents (B') As stated above, the compounds of this specification further comprise group B'. In some embodiments, B' is a therapeutic agent or an imaging agent (or a radical of either of the above). In some embodiments, B' is a radical of an anticancer agent or comprises the same. In some embodiments, B' is a radical of a dye (e.g., a fluorescent dye) or comprises the same. In some embodiments, B' is a radical of an antifibrotic agent or comprises the same. In some embodiments, B' is a radical of a PI3K inhibitor or comprises the same. In some embodiments, B' is a chelated radioisotope (e.g., 99m Tc, 111 In, 18 F, 68 Ga, 124 I, 125 I, 131 I, 32 P, 89 Sr, 90 Y, 153 Sm, 169 Er, 177 Lu, 186 Re, 188 Re, 149 Tb, 211 At, 212 Bi, 213 Bi or 225 A radical of a radiation imaging agent containing Ac) or containing this. B' is, 99m Tc, 111 In, 18 F, 68 Ga, 124 I, 125 I and 131It may be a radical of a radioactive isotope selected from the group consisting of I, or it may contain such a radical. In a particular embodiment, B' is 32 P, 89 Sr, 90 Y, 153 Sm, 169 Er, 177 Lu, 186 Re, 188 Re, 149 Tb, 211 At, 212 Bi, 213 Bi and 225 A radioactive isotope radical selected from the group consisting of Ac, or containing such a radical.

[0323] In certain embodiments, B' is a PI3K inhibitor, a radical of a chelate group optionally bound to an isotope (or metal), or a group covalently bound to an isotope (or metal), wherein the isotope or metal is suitable for radioimaging, radiotherapy, or magnetic resonance imaging; anticancer agents; antifibrotic agents; and / or dyes (e.g., fluorescent dyes).

[0324] In some embodiments, B' is a radical of a chelate group optionally bonded to an isotope (or metal) or a group covalently bonded to an isotope (or metal), wherein the isotope (or metal) is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0325] In some embodiments, B' is a radical of a radioimaging agent, a radiotherapy isotope or a magnetic resonance isotope, or a chelating group, and a radical of a radioimaging agent, a radiotherapy isotope or a magnetic resonance isotope bonded to the chelating group, or comprising these, wherein the isotope is 32 P, 89 Sr, 90 Y, 153 Sm, 169 Er, 177 Lu, 186 Re, 188 Re, 149 Tb, 211 At,212 Bi, 213 Bi and 225 Selected from the group consisting of Ac. In some embodiments, B' is a radical of a radioimaging agent, a radiotherapy isotope or a magnetic resonance isotope, or a chelating group, and a radical of a radioimaging agent, a radiotherapy isotope or a magnetic resonance isotope bonded to the chelating group, or comprising these, wherein the isotope is 18 F, 32 P, 44 Sc, 47 Sc, 52 Mn, 55 Co, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 86 Y, 89 Sr, 89 Zr, 90 Y, 99m Tc, 111 In, 114m In, 117m Sn, 124 I, 125 I, 131 I, 149 Tb, 153 Sm, 152 Tb, 155 Tb, 161 Tb, 169 Er, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 224 Ra, 225 Ab, 225 Ac or 227 Selected from Th. In some embodiments, the isotope is 111 In some embodiments, the isotope is 177 It is Lu. In some embodiments, the isotope is 11 C, 13 C, 13 N, 15 O, 60 Co and 123Selected from the group consisting of I.

[0326] The therapeutic agent (or its radical) may be any entity capable of producing a desired physiological response. The therapeutic agent (or radical) may be an antifibrotic agent, an anticancer agent, a chemotherapeutic agent, a photodynamic agent, a radiotherapeutic agent, etc. The therapeutic agent may be a compound (e.g., or its radical) that is effective against cancer cells or pro-fibrotic cells (e.g., cancer-associated fibroblasts, myofibroblasts, etc. (e.g., other tumor microenvironment factors)) (e.g., effective in eliminating, destroying, reducing (e.g., reducing the number of those cells), or mitigating the effects of those cells). Examples of therapeutic agents (or their radicals) include, but are not limited to, photodynamic agents, radiotherapeutic agents, chemotherapeutic agents, antifibrotic agents, and anticancer agents. The therapeutic agents provided herein may be PI3K inhibitors (or their radicals). Figure 20A shows the structures of various exemplary FAP5-PI3K inhibitors.

[0327] The therapeutic agent may be an anticancer agent (or its radical). The therapeutic agent may be an antifibrotic agent (or its radical). The therapeutic agent may be a compound (or its radical) selected from tumor growth factor (TGF) β / Smad inhibitors, Wnt / β-catenin inhibitors, kinase inhibitors (e.g., kinase inhibitors for vascular endothelial growth factor receptor (VEGFR), kinase inhibitors for fibroblast growth factor receptor (FGFR), kinase inhibitors for platelet-derived growth factor receptor (PDGFR), kinase inhibitors for focal adhesion kinase (FAK), or kinase inhibitors for Rho-related protein kinase (ROCK)), Toll-like receptor agonists (TLRs), nuclear factor kappa-light chain enhancer (NF-κB) as an activated B cell inhibitor, collagen synthesis inhibitors, and PI3K inhibitors. In certain embodiments, B' is a phosphoinositide-3-kinase (PI3K) inhibitor (or its radical).

[0328] In a particular embodiment, B' is

[0329] [ka] It is selected from the group consisting of the following.

[0330] In certain embodiments, B' is an imaging agent. The imaging agent may be any compound (or its radical) that emits a detectable signal (e.g., an electromagnetic signal (e.g., a radio signal, a fluorescence signal, a gamma ray) or mass). Examples of imaging agents include, but are not limited to, radiation imaging agents (e.g., PET imaging agents or SPECT imaging agents), fluorescence imaging agents (e.g., fluorescent dyes), etc. The imaging agent may also be a magnetic resonance (MR) agent. In some embodiments, B' comprises a radiolabeled functional group (e.g., its radical) suitable for PET imaging, SPECT imaging, other radiation imaging techniques, magnetic resonance imaging, or radiotherapy. B' may include radicals of radiation imaging isotopes, radiotherapy isotopes, or magnetic resonance isotopes.

[0331] B' may include imaging agents, radioimaging agents, photodynamic agents, chemotherapeutic agents, antifibrotic agents and / or radiotherapeutic agents (e.g., their radicals), and B' is an anticancer agent effective against cancer cells or cancer-associated fibroblasts, myofibroblasts or other tumor microenvironment factors.

[0332] B' may be a radical of a PI3K inhibitor. B' may be a radical of a chelate group optionally bound to an isotope (or metal). B' may be a chelate group covalently bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging.

[0333] B' may be a chelating group (e.g., a chelating agent (or its radical)). Typical chelating groups include, but are not limited to, their free bases, such as those formed by the removal of one or more protons (H+) from CO2H(COOH) to form COO-.

[0334] [ka] Includes, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0335] B' may contain a chelating group, which is not limited to, DOTA(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) or its derivatives; TETA(1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid) or its derivatives; SarAr(1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]-eicosane-1,8-diamine) or its derivatives; NOTA(1,4,7-Triazacyclononane-1,4,7-triacetic acid) or its derivatives; NETA(4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl)acetic acid or its derivatives; TRAP(1,4,7-Triazacyclononane-1,4,7-tris[methyl(2-carboxyethyl)phosphinic acid) or its derivatives; HBED(N,N0-Bis(2-hydroxybenzyl)-ethylenediamine-N ,N0-diacetic acid) or its derivatives; 2,3-HOPO(3-hydroxypyridine-2-one) or its derivatives; PCTA(3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-1(15),11,13-triene-3,6,9-triacetic acid) or its derivatives; DFO(desferrioxamine) or its derivatives; DTPA(diethylenetriaminepentaacetic acid) or its derivatives; OCTAPA(N,N0-bis(6-carboxy-2-pyridylmethyl)- The EC20 head includes ethylenediamine-N,N0-diacetic acid) or its derivatives; or H2-MACROPA(N,N'-bis[(6-carboxy-2-pyridylmethyl (pyridipmethyl)]-4,13-diaza-18-crown-6) or its derivatives; H2dedpa(1,2-[[carboxy)-pyridine-2-yl]-methylamino]ethane or its derivatives; and β-1-diaminopropionic acid, aspartic acid, and cysteine.

[0336] B' can contain a radical of DOTA. In certain embodiments, B' is isotope-(or metal-)chelated DOTA(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid).

[0337] B' may be, or may contain, a radical of a group covalently bonded to an isotope (or metal) suitable for radioimaging, radiotherapy, or magnetic resonance imaging.

[0338] B' may be, or may contain, a chelate group bound to an isotope (or metal) suitable for PET imaging, SPECT imaging, other radiographic imaging techniques, or radiotherapy.

[0339] B' is,

[0340] [ka] That's fine.

[0341] B' is optionally bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging.

[0342] [ka] That's fine.

[0343] B' is the following chelating group

[0344] [ka] It may be one of these, each optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0345] B' may contain magnetic resonance, radiotherapy, or radiotherapeutic isotopes. In some embodiments, B' includes a chelate group and a radiotherapy isotope, radiotherapeutic isotope, or magnetic resonance isotope, which is a metal (e.g., a metal suitable for radiotherapy, radiotherapy, or magnetic resonance imaging) bonded to the chelate group. In some embodiments, the isotope is a metal atom bonded to the chelate group of B'. In some embodiments, the radiotherapy isotope, radiotherapeutic isotope, or magnetic resonance isotope (or a metal suitable for radiotherapy, radiotherapy, or magnetic resonance imaging) is 18 F, 32 P, 44 Sc, 47 Sc, 52 Mn, 55 Co, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 86 Y, 89 Sr, 89 Zr, 90 Y, 99m Tc, 111 In, 114m In, 117m Sn, 124 I, 125 I, 131 I, 149 Tb, 153 Sm, 152 Tb, 155 Tb, 161 Tb, 169 Er, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 224 Ra, 225 Ab, 225 Ac or 227 Th is used. In some embodiments, the radiation imaging isotope, the radiotherapy isotope, or the magnetic resonance isotope is 11 C, 13 C, 13 N, 15 O,60 Co or 123 I. In some embodiments, the radiation imaging isotope, the radiation therapy isotope, or the magnetic resonance isotope is 225 Ac, 32 P, 89 Sr, 117m Sn, 153 Sm, 169 Er, 186 Re, 188 Re, 149 Tb, 212 Bi or 213 It is Bi. In some embodiments, the isotope is 111 In some embodiments, the isotope is 177 It is Lu.

[0346] B' may contain a radioimaging nuclide. The radioimaging nuclide may be any suitable radioimaging nuclide. 99m Tc, 111 In, 18 F, 68 Ga, 124 I, 125 I and 131 You can choose from the group consisting of I.

[0347] B' may contain radionuclides for radiotherapy. Radionuclides for radiotherapy are, 177 Lu, 90 Y and 211 You can choose from the group consisting of At.

[0348] B' may contain a radiolabeled prosthetic group (or its radical). The radiolabeled prosthetic group is 18 F, 124 I, 125 I, 131 I and 211 It may contain radioactive isotopes selected from the group consisting of At.

[0349] In certain embodiments, B' (e.g., a radiolabeled prosthetic group (or its radical)) has the following structure:

[0350] [ka] It has, During the ceremony, Each X is independent of the others. 18 F, 124 I, 125 I, 131 I and 2ll It is a radioactive isotope selected from the group consisting of At, R or R 1 Each of these is independently H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, aryl, substituted aryl, heteroaryl, or substituted heteroaryl. Each n is an integer independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0351] Typical radiolabeled prosthetic groups (e.g., B') are, but are not limited to, the following:

[0352] [ka] Includes.

[0353] In the case of radionuclides used for radiotherapy, if B' is a chelating agent, in certain embodiments, B' can form a chelate with the radionuclide.

[0354] B' may be a PI3K inhibitor or its radical.

[0355] A PI3K inhibitor (or its radical) (for example, a compound or a conjugate containing a PI3K inhibitor (or its radical)) has the structure of formula III:

[0356] [ka] It can have, During the ceremony, X is

[0357] [ka] It is selected from the group consisting of the following.

[0358] In formula (III), X may be a radical of B' (for example, the radical may be located on a heteroatom (e.g., S, N, or O of X)). B' may be bonded to L of the compound via X (e.g., a hydroxyl radical of X).

[0359] A PI3K inhibitor (or its radical) of compound B' (e.g., a compound or a conjugate containing a PI-3 kinase inhibitor (or its radical)) has the following structure:

[0360] [ka] It can have.

[0361] L is

[0362] [ka] It may also be the case that n is an integer from 1 to 32. L is

[0363] [ka] It may also be, and m is an integer from 1 to 9. L is,

[0364] [ka] It may also be the case that m is an integer from 1 to 9, n is an integer from 1 to 32, q is an integer from 0 to 4, and s is an integer from 0 to 4.

[0365] m may be 1. m may be 2. m may be 3. m may be 4. m may be 5. m may be 6. m may be 7. m may be 8. m may be 9.

[0366] n can be 1 through 12. n can be 1. n can be 2. n can be 3. n can be 4. n can be 5. n can be 6. n can be 7. n can be 8. n can be 9. n can be 10. n can be 11. n can be 12. n can be 13. n can be 14. n can be 15. n can be 16. n can be 17. n can be 18. n can be 19. n can be 20. n can be 21. n can be 22. n can be 23. n can be 24. n can be 25. n can be 26. n can be 27. n can be 28. n can be 29. n can be 30. n can be 31. n can be 32.

[0367] q may be 0. q may be 1. q may be 2. q may be 3. q may be 4.

[0368] s may be 0. s may be 1. s may be 2. s may be 3. s may be 4.

[0369] Linker (L) L of this compound is a linker, such as any suitable linker. As used herein, the term “linker” generally refers to a portion of a compound that forms a chemical bond with A (e.g., a binding ligand) and / or B' (e.g., a therapeutic or imaging agent) and / or C' (e.g., an albumin-binding ligand, PEG, a peptide, a peptidoglycan, or a sugar). In particular, a “linker” can link two or more functional portions of a molecule to form the compounds provided herein. For example, a linker may contain 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 link different functional capabilities of a compound, such as the groups of an FAP ligand and a DOTA chelator. A linker may contain several linker groups, such as in the range of about 2 to about 100 atoms in a continuous main chain.

[0370] The linker may be a release linker. The linker may also be a non-release linker. L may be a trivalent linker. L may be a biofunctionalized linker. For example, in at least formulas (X) and (I), the linker (e.g., difunctionalized) can form a chemical bond between A and B'. L may be a linker (e.g., difunctionalized) that links one or more A groups to B' (e.g., by a first covalent bond linking L to A and a second covalent bond linking L to B').

[0371] L may contain one or more linker groups, each independently selected from the group consisting of alkyl (alkylene), heteroalkyl (alkylene), heterocycloalkyl (alkylene), heteroaryl, aryl, alkoxy, thioether, disulfide, carboxylic acid, anhydride, carbonate, carbamate, thioether, sugar, peptide, and peptidoglycan. L may contain one or more linker groups, each independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, anhydride, carbonate, ester, carbamate, thioether, triazole, sugar, and peptide. L may contain one or more linker groups, each independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, carbonate, ester, phenyl, triazole, and carbamate. L may contain one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, phenyl, triazole, ester, and carbonate. L may contain one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, ester, and carbonate. L may contain one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, and amide. L may contain one or more linker groups, each of which is independently selected from the group consisting of alkyl (alkylene), disulfide, and amide. L may contain one or more linker groups, each of which is independently selected from the group consisting of amide, alkyl (alkylene), PEG, phenyl, and triazole. L may contain one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), and amide.L may contain one or more linker groups, each independently selected from the group consisting of alkyl (alkylene) and amide. L may contain one or more linker groups, each independently selected from the group consisting of PEG and amide. The linker may contain one or more triazole linker groups. The linker may contain one or more todisulfide linker groups. The linker may contain one or more amide linker groups. The linker may contain one or more PEG linker groups.

[0372] L may be a non-releasing linker (for example, divalent and bonded to B' and A (for example, by covalent bonds)). L may be a releasing linker (for example, divalent and bonded to B' and A (for example, by covalent bonds)).

[0373] In certain embodiments, for example and not limited to, in formulas (I') and (II), L comprises three linker groups, each independently selected from the group consisting of alkyl(alkylene), heteroalkyl(alkylene), heterocycloalkyl(alkylene), heteroaryl, aryl, alkoxy, thioether, disulfide, carboxylic acid, anhydride, carbonate, carbamate, thioether, sugar, and peptide. In certain embodiments, L comprises three linker groups, each independently selected from the group consisting of PEG, alkyl(alkylene), disulfide, amide, carboxylic acid, anhydride, carbonate, ester, carbamate, thioether, triazole, sugar, and peptide. L may also comprise three linker groups, each independently selected from the group consisting of PEG, alkyl(alkylene), disulfide, amide, carboxylic acid, carbonate, ester, phenyl, triazole, and carbamate. In certain embodiments, L comprises three linker groups, each independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, phenyl, triazole, ester, and carbonate. L can comprise three linker groups, each independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, amide, carboxylic acid, ester, and carbonate. L can comprise three linker groups, each independently selected from the group consisting of PEG, alkyl (alkylene), disulfide, and amide. L can comprise three linker groups, each independently selected from the group consisting of alkyl (alkylene), disulfide, and amide. L can comprise three linker groups, each independently selected from the group consisting of amide, alkyl (alkylene), PEG, phenyl, and triazole. L can contain three linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), and amide.L can contain three linker groups, each of which is independently selected from the group consisting of alkyl (alkylene) and amide.

[0374] L may contain one or more release groups.

[0375] L can be bonded to A (for example, by covalent bonding) via an amide linker group. L can be bonded to B' (for example, by covalent bonding) via an amide linker group. L can be bonded to C' (for example, by covalent bonding) via an amide linker group. L can be bonded independently (by covalent bonding, etc.) to A, B' and (in formulas (I') and (II)) C' via an amide linker group.

[0376] L can be bonded to A (for example, by covalent bonding) via a triazole linker group. L can be bonded to B' (for example, by covalent bonding) via a triazole linker group. L can be bonded to A and B' independently (by covalent bonding, etc.) via a triazole linker group.

[0377] L can be bonded to A (for example, by covalent bonding) via a triazole linker group. L can be bonded to A via a triazole linker group and to B' via an amide linker group.

[0378] L can be bonded to B' (for example, by covalent bonding) via a carbamate linker group. L can be bonded to A via an amide linker group, and can be bonded to B' via a carbamate linker group.

[0379] A linker can be an ejected linker. A linker can be a non-ejected linker.

[0380] The linker may be a reductively cleavable linker (e.g., a disulfide). The linker may be an oxidatively cleavable linker (e.g., an amino-aromatic group). The linker may be an oxime ester or may contain one. The linker may be a hydrazone or may contain one. The linker may be a PEG n (n=0~36) may or may include this. The linker may or may include a peptide. The linker may or may include a peptidoglycan.

[0381] L is (L 1 )-(L 2 ) or (L 1 )-(L 2 )-(L 3 ) may also be, and in the formula, L 1 This is the first linker, L 2 It is the second linker, (If necessary), L 3 This is the third linker.

[0382] L 1 , L 2 and L 3 L may be the same. 1 , L 2 and L 3 Two of them may be the same. 1 , L 2 and L 3 They can be different. L 1 is the A group (and L 2 It can be connected to the base. 2 The B' group (and L 2 and L 3 It can bond to the base. 3 The C' group (and L 2 It can be bonded to the base.

[0383] L 1 , L2 and L 3 Each of these may independently have a length of 15 to 200 angstroms (Å).

[0384] L is (L 1 ) p -W-(L 2 ) q It may be so, and in the formula, L 1 This is the first linker, L 2 It is the second linker, p = 1 to 5, q = 1 to 5.

[0385] In a particular embodiment, each L 1 and each L 2 It independently comprises one or more linker groups, each of which is independently selected from the group consisting of PEG, alkyl (alkylene), amide, phenyl, and triazole.

[0386] W (if present) may be an amine core, an aromatic core, or an alkylene core.

[0387] L 1 and L 2 Each of these can be independent and have a length of 5 to 200 Å.

[0388] L has the following structure:

[0389] [ka] It can have.

[0390] L is

[0391] [ka] That's fine.

[0392] L is

[0393] [ka] That's fine.

[0394] L may contain at least one linker group, each of which is selected from the group consisting of PEG, alkyl, sugar, and peptide. The linker may be a dual linker based on PEG (e.g., PEGylated), alkyl, sugar, and peptide.

[0395] L may be a non-releasing linker (for example, divalent and bonded to B' and A (for example, by covalent bonds)). L may be a releasing linker (for example, divalent and bonded to B' and A (for example, by covalent bonds)).

[0396] L has the following structure:

[0397] [ka] It may contain one or more linker groups having the following characteristics: In the formula, n is between 0 and 10.

[0398] L has the following structure:

[0399] [ka] Having one or more linker groups (e.g., L 1 , L 2 and / or L 3 (where applicable) may include, In the formula, n is between 0 and 10.

[0400] L has the following structure:

[0401] [ka] It may contain one or more linker groups having the following characteristics: In the formula, n is between 1 and 32.

[0402] L has the following structure:

[0403] [ka] It may contain one or more linker groups having the following characteristics: In the formula, n is between 1 and 32.

[0404] L has the following structure:

[0405] [ka] It may contain one or more linker groups having the following characteristics: During the ceremony, R 12 and R 13 Each of these may independently be H or a C1-C6 alkyl group. z is an integer between 1 and 8.

[0406] L has the following structure:

[0407] [ka] It may contain one or more linker groups having the following characteristics: During the ceremony, R 12 and R 13 Each of these may independently be H or a C1-C6 alkyl group. z is an integer between 1 and 8.

[0408] L has the following structure:

[0409] [ka] Having one or more linker groups (e.g., L 1and each L 2 ) can include

[0410] L has the following structure:

[0411] [ka] It may contain one or more linker groups having

[0412] L has the following structure:

[0413] [ka] It may contain one or more linker groups having the following characteristics: During the ceremony, R 16 is H or C1-C6 alkyl, R 14a , R 14b , R 15a and R 15b Each of these may independently be H or a C1-C6 alkyl group.

[0414] L has the following structure:

[0415] [ka] It can include...

[0416] L has the following structure:

[0417] [ka] It may contain one or more linker groups having the following characteristics: In the formula, n is between 0 and 15.

[0418] L may contain a reductively cleavable linker. L may contain an oxidatively cleavable linker. L may contain an oxime ester. L may contain a hydrazone. L may contain PEGn, where n=0-36. L may contain a peptide. L may contain a peptidoglycan.

[0419] L is

[0420] [ka] That's fine.

[0421] L is

[0422] [ka] That's fine.

[0423] C’ In certain embodiments of the compounds described herein (for example, compounds having the structure of formula (I') or (II)), the compound further comprises a C' linked to the linker of the compound.

[0424] C' may be any pharmacokinetic extender. In certain embodiments, C' is an albumin-binding ligand. C' may be an albumin-binding small protein scaffold (ABD035) containing an albumin-binding domain 035, an albumin-binding domain Con (ABDCon) which is a peptide of 45 three-helix bundle amino acids, an engineered ankyrin repeat protein (DARPin), a disulfide-stabilized Fv fragment (dsFv)CA645 (anti-albumin antibody), any nanobody that forms a complex with human serum albumin (nanobody), and a variable new antigen receptor E06 (VNAR(E06)).

[0425] In a particular embodiment, C' is

[0426] [ka] It is either or includes this.

[0427] In a particular embodiment, C' is

[0428] [ka] is or includes the same, Where applicable, R 12~19 Each of these (as appropriate) independently corresponds to -H, -C1~C6 alkyl, -F, -Cl, -Br, -I, -CN, -CHO, -B(OH)2, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, or -SO2F. R 20 and R 21 These are independently -H, -C1~C6 alkyl, -F, -Cl, -Br, -I, -OC 1~6 These are alkyl, -CN, -CHO, -B(OH)2, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, -SO2F, or CF3.

[0429] In some embodiments, C' is

[0430] [ka] It is either or includes this.

[0431] In some embodiments, C' is

[0432] [ka] It is either or includes this.

[0433] In some embodiments, C' is

[0434] [ka] It is either or includes this.

[0435] In certain embodiments, the C' of the compound is (PEG) n (where n is an integer between 0 and 32), it includes peptides, peptidoglycans, or sugars (e.g., radicals of these).

[0436] C' is (PEG) n It may also be a peptidoglycan, where n is an integer between 0 and 32. C' may be a peptide. C' may be a peptidoglycan. C' may be a sugar.

[0437] In some embodiments, C' is

[0438] [ka] It is either or includes this.

[0439] In some embodiments, C' is

[0440] [ka] It is either or includes this.

[0441] Similarly, compounds of formula (V) (e.g., conjugates):

[0442] [ka] Provided During the ceremony, L is a linker containing at least one carbon atom, p is 0, 1, 2, or 3.

[0443] In the case of the compound of formula (V), L may be the group shown in one of the embodiments previously shown with respect to linker L, or may contain this group. In certain embodiments relating to the compound of formula (V), p may be 0. In certain embodiments relating to the compound of formula (V), p may be 1. In certain embodiments relating to the compound of formula (V), p may be 2. In certain embodiments relating to the compound of formula (V), p may be 3.

[0444] Compounds / Conjugates In some embodiments, the compound is as follows:

[0445] [ka] isn't it.

[0446] AL-B' has the following structure:

[0447] [ka] It can have, During the ceremony, n is an integer between 1 and 5.

[0448] AL-B' has the following structure:

[0449] [ka]

[0450] [ka] It can have, During the ceremony, n is an integer between 1 and 5.

[0451] This compound has the formula:

[0452] [ka] It can have, t is either 0 or 1, and u is an integer between 2 and 12.

[0453] This compound is, formula

[0454] [ka] It can have such a value, where n is an integer from 1 to 12.

[0455] This compound is, formula

[0456] [ka] It can have m, where m is an integer from 1 to 4.

[0457] This compound is, formula

[0458] [ka] It can have.

[0459] This compound is, formula

[0460] [ka] It can have.

[0461] This compound is, formula

[0462] [ka] It can have.

[0463] This compound is, formula

[0464] [ka] It can have.

[0465] This compound is, formula

[0466] [ka] It can have.

[0467] In certain embodiments, the AL-B' of the compounds herein has the following structure:

[0468] [ka] It can have.

[0469] AL-B' has the following structure:

[0470] [ka] It can have.

[0471] AL-B' has the following structure:

[0472] [ka] It can have.

[0473] AL-B' has the following structure:

[0474] [ka] It can have.

[0475] AL-B' has the following structure:

[0476] [ka]

[0477] [ka] It can have.

[0478] AL-B' is a structure that may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, as follows:

[0479] [ka] It can have.

[0480] AL-B' is a structure that may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, as follows:

[0481] [ka] It can have.

[0482] AL-B' is a structure that may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, as follows:

[0483] [ka] It can have.

[0484] AL-B' is a structure that may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, as follows:

[0485] [ka] It can have.

[0486] AL-B' is a structure that may be bound to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, as follows:

[0487] [ka] It can have.

[0488] The compound (e.g., conjugate) has the following structure:

[0489] [ka]

[0490] [ka] It can have.

[0491] The compound (e.g., conjugate) has the following structure:

[0492] [ka]

[0493] [ka] It can have.

[0494] The compound (e.g., conjugate) has the following structure:

[0495] [ka]

[0496] [ka] It can have.

[0497] The compound (e.g., conjugate) has the following structure:

[0498] [ka]

[0499] [ka] It can have.

[0500] The compound (e.g., conjugate) has the following structure:

[0501] [ka] It can have.

[0502] The compound (e.g., conjugate) has the following structure:

[0503] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0504] The compound (e.g., conjugate) has the following structure:

[0505] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0506] The compound (e.g., conjugate) has the following structure:

[0507] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0508] The compound (e.g., conjugate) has the following structure:

[0509] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0510] The compound (e.g., conjugate) has the following structure:

[0511] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0512] The compound (e.g., conjugate) may be conjugated to an isotope (or metal) suitable for radiographic imaging, radiotherapy, or magnetic resonance imaging, and may have the following structure:

[0513] [ka] It can have.

[0514] The compound (e.g., conjugate) has the following structure:

[0515] [ka]

[0516] [ka]

[0517] [ka] It can have.

[0518] The compound (e.g., conjugate) has the following structure:

[0519] [ka]

[0520] [ka]

[0521] [ka] It can have.

[0522] The compound (e.g., conjugate) has the following structure:

[0523] [ka]

[0524] [ka]

[0525] [ka] It can have.

[0526] The compound (e.g., conjugate) has the following structure:

[0527] [ka]

[0528] [ka]

[0529] [ka] It can have.

[0530] The compound (e.g., conjugate) has the following structure:

[0531] [ka]

[0532] [ka]

[0533] [ka] It can have.

[0534] The compound (e.g., conjugate) has the following structure:

[0535] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0536] The compound (e.g., conjugate) has the following structure:

[0537] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0538] The compound (e.g., conjugate) has the following structure:

[0539] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0540] The compound (e.g., conjugate) has the following structure:

[0541] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0542] The compound (e.g., conjugate) has the following structure:

[0543] [ka] It can have, Each is optionally bound to an isotope (or metal) suitable for radiation imaging, radiation therapy, or magnetic resonance imaging.

[0544] This compound can have any of the structures shown in Figure 1, each optionally bound to an isotope (or metal) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

[0545] This compound has the formula:

[0546] [ka] It can have, t is either 0 or 1, and u is an integer between 2 and 12.

[0547] This compound is, formula

[0548] [ka] It can have such a value, where n is an integer from 1 to 12.

[0549] This compound is, formula

[0550] [ka] It can have m, where m is an integer from 1 to 4.

[0551] This compound is, formula

[0552] [ka] It can have.

[0553] This compound is, formula

[0554] [ka] It can have.

[0555] This compound is, formula

[0556] [ka] It can have.

[0557] This compound is, formula

[0558] [ka] It can have.

[0559] This compound is, formula

[0560] [ka] It can have.

[0561] This compound may have any of the formulas shown in Figure 20A.

[0562] In a particular embodiment, the compound (i.e., the conjugate) has the following structure:

[0563] [ka] It can have.

[0564] In a particular embodiment, the compound (i.e., the conjugate) has the following structure:

[0565] [ka] It can have.

[0566] In a particular embodiment, the compound (i.e., the conjugate) has the following structure:

[0567] [ka] It can have.

[0568] In a particular embodiment, the compound (i.e., the conjugate) has the following structure:

[0569] [ka] It can have.

[0570] In certain embodiments, the compound (i.e., the conjugate) may have any of the structures shown in Figures 28-31.

[0571] In a particular embodiment, the compound has the following structure:

[0572] [ka] It may have any of the following:

[0573] Pharmaceutical composition, route of administration and administration In certain embodiments, a pharmaceutical composition comprising a compound and a pharmaceutically acceptable carrier is provided. In certain embodiments, the pharmaceutical composition comprises a plurality of compounds and a pharmaceutically acceptable carrier.

[0574] In certain embodiments, the pharmaceutical composition further comprises at least one additional pharmaceutically active agent. The at least one additional pharmaceutically active agent may be an agent useful for treating ischemia-reperfusion injury.

[0575] Pharmaceutical compositions can be prepared by combining one or more compounds with a pharmaceutically acceptable carrier and, optionally, one or more additional pharmaceutically active agents.

[0576] As used herein, “effective dose” refers to any amount sufficient to achieve the desired biological effect. By combining the teachings presented herein with a selection of various active compounds and weighting factors such as potency, relative bioavailability, patient weight, severity of adverse side effects, and mode of administration, it is possible to design effective prophylactic or therapeutic regimens that do not cause serious undesirable toxicity but remain effective in treating a particular subject. The effective dose for any particular use may vary depending on factors such as the disease or condition being treated, the specific compound administered, the size of the subject, or the severity of the disease or condition. Those skilled in the art can empirically determine the effective dose of a particular compound and / or other therapeutic agent without requiring excessive experimentation. Depending on some medical judgment, the maximum dose, i.e., the safest dose, may be used. A repeated daily dose can be used to achieve an appropriate systemic level of the compound. An appropriate systemic level can be determined, for example, by a patient’s peak measurement or by a sustained plasma level of the drug. “Dose” and “administration” are used interchangeably herein.

[0577] Generally, the daily oral dose of a compound in human subjects ranges from approximately 0.01 mg / kg to 1,000 mg / kg per day. Oral doses ranging from 0.5 to 50 mg / kg in one or more daily doses can produce therapeutic effects. Dosages can be appropriately adjusted to achieve the desired local or systemic drug level, depending on the mode of administration. For example, intravenous administration can vary in doses that are one or even several orders of magnitude lower per day. If the response in a subject is insufficient at such doses, higher doses (or higher doses with greater efficacy via a different, more localized delivery route) can be used up to the extent that the patient can tolerate. Repeated daily doses are intended to achieve an appropriate systemic level of the compound.

[0578] With respect to use in treatment, the “therapeutic effective dose” (or “effective dose”) of a compound means the amount of the compound in a preparation that, when administered (to a mammal such as a human) as part of a desired administration regimen, reduces symptoms, improves a condition, or delays the onset of a disease condition, in accordance with clinically acceptable standards for the disorder or condition being treated or for cosmetic purposes, for example, in a reasonable benefit / risk ratio applicable to any medical treatment.

[0579] For any given compound, the therapeutically effective dose can first be determined from animal models. The therapeutically effective dose can also be determined from human data for compounds tested in humans, and for compounds known to exhibit similar pharmacological activity, such as other related activators. Higher doses may be required for parenteral administration. The applicable dose can be adjusted based on the relative bioavailability and potency of the compound being administered. Based on the above methods and other methods well known in the art, dose adjustments to achieve maximum potency are well within the capabilities of those skilled in the art.

[0580] With regard to clinical use, each compound may be administered in an amount equivalent to 0.2 to 2,000 milligrams (mg) of the compound per kilogram (kg) of the subject's body weight per day. This compound may be administered in an amount equivalent to 2 to 2,000 mg of the compound per kg of the subject's body weight per day. This compound may be administered in an amount equivalent to 20 to 2,000 mg of the compound per kg of the subject's body weight per day. This compound may be administered in an amount equivalent to 50 to 2,000 mg of the compound per kg of the subject's body weight per day. This compound may be administered in an amount equivalent to 100 to 2,000 mg of the compound per kg of the subject's body weight per day. This compound may be administered in an amount equivalent to 200 to 2,000 mg of the compound per kg of the subject's body weight per day. If a precursor or prodrug of the compound is to be administered, it will be administered in an equivalent amount, i.e., a sufficient amount, to deliver the above-mentioned amount of the compound.

[0581] Compound formulations can be administered to human subjects in therapeutically effective doses. Typical dose ranges are approximately 0.01 micrograms to 2 mg per kg of body weight per day. The dosage of a drug administered is likely to depend on variables such as the type and severity of the disorder, the overall health status of the particular subject, the specific compound administered, the excipients used to formulate the compound, and its route of administration. Conventional experiments can be used to optimize the dose and frequency of administration for any particular compound.

[0582] This compound can be administered at concentrations ranging from approximately 0.001 micrograms / kg to over 500 mg / kg. For example, concentrations can be 0.001 micrograms / kg, 0.01 micrograms / kg, 0.05 micrograms / kg, 0.1 micrograms / kg, 0.5 micrograms / kg, 1.0 micrograms / kg, 10.0 micrograms / kg, 50.0 micrograms / kg, 100.0 micrograms / kg, 500 micrograms / kg, 1.0 mg / kg, 5.0 mg / kg, 10.0 mg / kg, 15.0 mg / kg, 20.0 mg / kg, 25.0 mg / kg, and 3. The values ​​may range from 0.0 mg / kg, 35.0 mg / kg, 40.0 mg / kg, 45.0 mg / kg, 50.0 mg / kg, 60.0 mg / kg, 70.0 mg / kg, 80.0 mg / kg, 90.0 mg / kg, 100.0 mg / kg, 150.0 mg / kg, 200.0 mg / kg, 250.0 mg / kg, 300.0 mg / kg, 350.0 mg / kg, 400.0 mg / kg, 450.0 mg / kg, up to approximately 500.0 mg / kg, or any increment thereof. It should be understood that all values ​​and ranges between these values ​​and ranges are intended to be included.

[0583] This compound can be administered in doses ranging from approximately 0.2 mg / kg / day to over 100 mg / kg / day. For example, doses include 0.2 mg / kg / day to 100 mg / kg / day, 0.2 mg / kg / day to 50 mg / kg / day, 0.2 mg / kg / day to 25 mg / kg / day, 0.2 mg / kg / day to 10 mg / kg / day, 0.2 mg / kg / day to 7.5 mg / kg / day, 0.2 mg / kg / day to 5 mg / kg / day, 0.25 mg / kg / day to 100 mg / kg / day, 0.25 mg / kg / day to 50 mg / kg / day, 0.25 mg / kg / day to 25 mg / kg / day, and 0.25 mg / kg / day to... 10mg / kg / day, 0.25mg / kg / day~7.5mg / kg / day, 0.25mg / kg / day~5mg / kg / day, 0.5mg / kg / day~50mg / kg / day, 0.5mg / kg / day~25mg / kg / day, 0.5mg / kg / day~ 20mg / kg / day, 0.5mg / kg / day ~ 15mg / kg / day, 0.5mg / kg / day ~ 10mg / kg / day, 0.5mg / kg / day ~ 7.5mg / kg / day, 0.5mg / kg / day ~ 5mg / kg / day, 0.75mg / kg / day ~ 50 mg / kg / day, 0.75mg / kg / day ~ 25mg / kg / day, 0.75mg / kg / day ~ 20mg / kg / day, 0.75mg / kg / day ~ 15mg / kg / day, 0.75mg / kg / day ~ 10mg / kg / day, 0.75mg / kg / day ~7.5mg / kg / day, 0.75mg / kg / day ~5mg / kg / day, 1.0mg / kg / day ~50mg / kg / day, 1.0mg / kg / day ~25mg / kg / day, 1.0mg / kg / day ~20mg / kg / day, 1.0mg / kg / day ~1 The dosage may be 5 mg / kg / day, 1.0 mg / kg / day to 10 mg / kg / day, 1.0 mg / kg / day to 7.5 mg / kg / day, 1.0 mg / kg / day to 5 mg / kg / day, 2 mg / kg / day to 50 mg / kg / day, 2 mg / kg / day to 25 mg / kg / day, 2 mg / kg / day to 20 mg / kg / day, 2 mg / kg / day to 15 mg / kg / day, 2 mg / kg / day to 10 mg / kg / day, 2 mg / kg / day to 7.5 mg / kg / day, or 2 mg / kg / day to 5 mg / kg / day.

[0584] This compound can be administered in doses ranging from approximately 0.25 mg / kg / day to approximately 25 mg / kg / day. For example, the doses are 0.25 mg / kg / day, 0.5 mg / kg / day, 0.75 mg / kg / day, 1.0 mg / kg / day, 1.25 mg / kg / day, 1.5 mg / kg / day, 1.75 mg / kg / day, 2.0 mg / kg / day, 2.25 mg / kg / day, 2.5 mg / kg / day, 2.75 mg / kg / day, 3.0 mg / kg / day, 3.25 mg / kg / day, 3.5 mg / kg / day, 3.75 mg / kg / day, and 4.0 mg / kg / day. g / day, 4.25mg / kg / day, 4.5mg / kg / day, 4.75mg / kg / day, 5mg / kg / day, 5.5mg / kg / day, 6.0mg / kg / day, 6.5mg / kg / day, 7.0mg / kg / day, 7. 5mg / kg / day, 8.0mg / kg / day, 8.5mg / kg / day, 9.0mg / kg / day, 9.5mg / kg / day, 10mg / kg / day, 11mg / kg / day, 12mg / kg / day, 13mg / kg / day, 14 mg / kg / day, 15mg / kg / day, 16mg / kg / day, 17mg / kg / day, 18mg / kg / day, 19mg / kg / day, 20mg / kg / day, 21mg / kg / day, 22mg / kg / day, 23mg / k g / day, 24mg / kg / day, 25mg / kg / day, 26mg / kg / day, 27mg / kg / day, 28mg / kg / day, 29mg / kg / day, 30mg / kg / day, 31mg / kg / day, 32mg / kg / day, 3 The dosage may be 3 mg / kg / day, 34 mg / kg / day, 35 mg / kg / day, 36 mg / kg / day, 37 mg / kg / day, 38 mg / kg / day, 39 mg / kg / day, 40 mg / kg / day, 41 mg / kg / day, 42 mg / kg / day, 43 mg / kg / day, 44 mg / kg / day, 45 mg / kg / day, 46 mg / kg / day, 47 mg / kg / day, 48 mg / kg / day, 49 mg / kg / day, or 50 mg / kg / day.

[0585] This compound or its precursor can be administered at concentrations ranging from 0.01 micromolar to over 500 micromolar. For example, doses can be 0.01 micromolar, 0.02 micromolar, 0.05 micromolar, 0.1 micromolar, 0.15 micromolar, 0.2 micromolar, 0.5 micromolar, 0.7 micromolar, 1.0 micromolar, 3.0 micromolar, 5.0 micromolar, 7.0 micromolar, 10.0 micromolar, 15.0 micromolar, 20.0 micromolar, 25.0 micromolar, 30.0 micromolar, 35.0 micromolar, and 40.0 micromolar. Micromolar concentrations may range from 45.0 micromolars, 50.0 micromolars, 60.0 micromolars, 70.0 micromolars, 80.0 micromolars, 90.0 micromolars, 100.0 micromolars, 150.0 micromolars, 200.0 micromolars, 250.0 micromolars, 300.0 micromolars, 350.0 micromolars, 400.0 micromolars, 450.0 micromolars, up to approximately 500.0 micromolars, or any increment thereof. It should be understood that all values ​​and ranges between these values ​​and ranges are intended to be included.

[0586] This compound or its precursor can be administered at concentrations ranging from 0.10 micrograms / mL to 500.0 micrograms / mL. For example, concentrations can be 0.10 micrograms / mL, 0.50 micrograms / mL, 1 microgram / mL, 2.0 micrograms / mL, 5.0 micrograms / mL, 10.0 micrograms / mL, 20 micrograms / mL, 25 micrograms / mL, 30 micrograms / mL, 35 micrograms / mL, 40 micrograms / mL, 45 micrograms / mL, 50 micrograms / mL, 60.0 micrograms / mL, and 70.0 micrograms / mL. L, 80.0 micrograms / mL, 90.0 micrograms / mL, 100.0 micrograms / mL, 150.0 micrograms / mL, 200.0 micrograms / mL, 250.0 g / mL, 250.0 micrograms / mL, 300.0 micrograms / mL, 350.0 micrograms / mL, 400.0 micrograms / mL, 450.0 micrograms / mL, up to approximately 500.0 micrograms / mL, or any increment thereof. It should be understood that all values ​​and ranges between these values ​​and ranges are intended to be included.

[0587] The formulation can be administered in a pharmaceutically acceptable solution, which may conventionally contain pharmaceutically acceptable concentrations of salts, buffers, preservatives, compatible carriers, adjuvants, and optionally other therapeutic components. For therapeutic use, an effective amount of the compound can be administered to a target by any means of delivering the compound to the desired surface. Administration of the pharmaceutical composition may be carried out by any means known to those skilled in the art. Routes of administration include, but are not limited to, intravenous, intramuscular, intraperitoneal, intravesical (bladder), oral, subcutaneous, direct injection (e.g., intratumor or abscess), mucosal (e.g., topical to the eye), inhalation, and topical.

[0588] For intravenous and other parenteral administration routes, the compound can be formulated as a lyophilized preparation, as a lyophilized preparation of the active compound intercalated in liposomes or encapsulated, as a lipid complex in an aqueous suspension, or as a salt complex. The lyophilized preparation is generally reconstituted immediately before administration in a suitable aqueous solution, such as sterile water or saline.

[0589] For oral administration, this compound can be readily formulated by combining the active compound with pharmaceutically acceptable carriers well known in the art. Such carriers allow the compound to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc., for oral administration, depending on the target of treatment. Pharmaceutical preparations for oral use can be obtained as solid excipients by optionally grinding the resulting mixture, adding suitable adjuvants if desired, and then processing the granular mixture to obtain the core of tablets or dragees. Suitable excipients include sugars, particularly lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or fillers such as polyvinylpyrrolidone (PVP). If desired, disintegrants such as cross-linked PVP, agar or alginate, or salts thereof such as sodium alginate may be added. Depending on the circumstances, oral formulations may also be formulated in physiological saline or a buffer to neutralize the internal acidity, such as EDTA, or they may be administered without any carrier.

[0590] Similarly, oral dosage forms of the compound are intended. The compound may be chemically modified so that oral delivery of the derivative is effective. Generally, the intended chemical modification involves the attachment of at least one moiety to the compound itself, in which case the moiety enables (a) inhibition of acid hydrolysis and (b) uptake into the bloodstream from the stomach or intestines. Similarly, increased overall stability of the compound and increased circulating time in the body are desirable. Examples of such moieties include polyethylene glycol, copolymers of ethylene glycol and propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, PVP, and polyproline. Abuchowski and Davis, "Soluble Polymer-Enzyme Adducts," In: Enzymes as Drugs, Hocenberg and Roberts, eds., Wiley-Interscience, New York, NY, pp. 367-383 (1981); Newmark et al., J Appl Biochem 4:185-189 (1982). Other polymers that can be used are poly-1,3-dioxolane and poly-1,3,6-thioxocan. For pharmaceutical use, the polyethylene glycol portion is preferred, as indicated above.

[0591] The release site of the compounds described herein may be the stomach, small intestine (e.g., duodenum, jejunum, or ileum), or large intestine. Those skilled in the art have available formulations that do not dissolve in the stomach but release the substance in the duodenum or somewhere in the intestine. This release can avoid adverse effects on the gastric environment by either protecting the compound or releasing the compound beyond the gastric environment, such as in the intestines.

[0592] To ensure complete resistance in the stomach, a coating that is impermeable to at least pH 5.0 is essential. Examples of more common inert ingredients used as enteric coatings include cellulose trimellitic acetate (CAT), hydroxypropyl methylcellulose phthalate (HPMCP), HPMCP50, HPMCP55, polyvinyl acetate phthalate (PVAP), Eudragit L30D, Aquateric, cellulose phthalate acetate (CAP), Eudragit L, Eudragit S, and shellac. These coatings can be used as mixed films.

[0593] Coating agents, or mixtures of coating agents, may also be used on the surface of tablets, not intended to protect the stomach. These may include sugar coating agents or coating agents that facilitate the swallowing of tablets. Capsules may consist of a hard shell (such as gelatin) for the delivery of dry therapeutic agents (e.g., powders), and a soft gelatin shell may be used for liquid forms. The shell material for cachets may be thick starch paper or other edible paper. For pills, lozenges, molded tablets, or powder tablets, the moist mashing technique may be used.

[0594] The therapeutic agent can be incorporated into the formulation as fine multiparticles in the form of granules, or as pellets with a particle size of approximately 1 mm. Formulations of the substance for capsule administration may also be lightly compressed plugs of powder, or even tablets. The therapeutic agent can be prepared by compression.

[0595] All flavorings and colorings may be included. For example, the compound can be formulated (e.g., by encapsulation in liposomes or microsphere capsules) and then further incorporated into edible products such as chilled beverages that also contain colorings and flavorings.

[0596] The volume of the therapeutic agent may be diluted or increased using an inert substance. These diluents may include carbohydrates, particularly mannitol, α-lactose, anhydrous lactose, cellulose, sucrose, modified dextran, and starch. Certain inorganic salts, including calcium triphosphate, magnesium carbonate, and sodium chloride, may also be used as fillers. Some commercially available diluents include Fast-Flo, Emdex, STA-Rx1500, Emcompress, and Avicell.

[0597] Disintegrants can be incorporated into the solid dosage form of therapeutic formulations. Substances used as disintegrants include, but are not limited to, starch, including commercially available starch-based disintegrants such as Explotab. Sodium starch glycolate, amberlite, sodium carboxymethylcellulose, ultraamylopectin, sodium alginate, gelatin, orange peel, acidic carboxymethylcellulose, sponge, and bentonite can all be used. Another form of disintegrant is an insoluble cation exchange resin. Powdered gums can be used as disintegrants and binders, and these may include powdered gums such as agar, karaya, or tragacanth. Alginic acid and its sodium salts are also useful as disintegrants.

[0598] Binders are used to bind therapeutic agents together to form hard tablets and include substances derived from natural products such as acacia, tragacanth, starch, and gelatin. Others include methylcellulose (MC), ethylcellulose (EC), and carboxymethylcellulose (CMC). Both PVP and hydroxypropyl methylcellulose (HPMC) can be used in alcoholic solutions to granulate therapeutic agents.

[0599] Antifriction agents can be incorporated into the formulation of the treatment agent to prevent adhesion during the formulation process. Lubricants can be used as a layer between the treatment agent and the die wall, and these may include, but are not limited to, stearic acid containing magnesium and calcium salts, polytetrafluoroethylene (PTFE), liquid paraffin, vegetable oils, and waxes. Soluble lubricants such as sodium lauryl sulfate, magnesium lauryl sulfate, polyethylene glycol of various molecular weights, and Carbowax 4000 and 6000 can also be used.

[0600] A flow enhancer can be added to improve the flow properties of the drug during formulation and to assist in rearrangement during compression. The flow enhancer may include starch, talc, calcined silica, and hydrated aluminosilicate.

[0601] To aid in the elution of therapeutic agents into an aqueous environment, surfactants may be added as wetting agents. Surfactants may include anionic detergents such as sodium lauryl sulfate, sodium dioctyl sulfosuccinate, and sodium dioctyl sulfonate. Cationic detergents that can be used include benzalkonium chloride and benzethonium chloride. Potentially nonionic detergents that may be included in the formulation as surfactants include lauromacrogol 400, polyoxyl stearate 40, polyoxyethylene hydrogenated castor oil 10, 50, and 60, glycerol monostearate, polysorbate 40, 60, 65, and 80, sucrose fatty acid esters, methylcellulose, and carboxymethylcellulose. These surfactants may be present in the formulation of the compound or its derivatives either individually or in mixtures of different ratios.

[0602] Pharmaceutical preparations that can be used orally include push-fit capsules made from gelatin, and soft, sealable capsules made from gelatin and a plasticizer such as glycerol or sorbitol. Push-fit capsules may contain the active ingredient in a mixture with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol. Further stabilizers may be added. Microspheres formulated for oral administration can also be used. Such microspheres are well defined in the art. All formulations for oral administration should be in a dosage suitable for such administration.

[0603] With regard to oral administration, this composition can conventionally take the form of a tablet or lozenge.

[0604] For local administration, the compound can be formulated as a solution, gel, ointment, cream, suspension, etc., as is well known in this art. Systemic formulations include those designed for administration by injection, such as subcutaneous, intravenous, intramuscular, intra-shelter, or intraperitoneal injection, and those designed for transdermal, transmucosal, oral, or pulmonary administration.

[0605] For administration by inhalation, the compound can be conveniently delivered in the form of an aerosol spray supply from a pressurized pack or nebulizer, along with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or other suitable gas. In the case of pressurized aerosols, the dose unit can be determined by providing a valve for delivering the measured amount. For use in inhalers or inhalation devices, formulations can be made, for example, of gelatin capsules and cartridges containing a powder mix of the compound and a suitable powder base such as lactose or starch.

[0606] Similarly, pulmonary delivery of the compound (or its salt) is attempted. The compound is delivered to the mammalian lungs during inhalation, passes through the pulmonary epithelial lining, and enters the bloodstream. Other reports on inhaled molecules include: Adjei et al., Pharm Res 7:565-569 (1990); Adjei et al., Int J Pharmaceutics 63:135-144 (1990) (leuprolide acetate); Braquet et al., J Cardiovasc Pharmacol 13(suppl. 5):143-146 (1989) (endothelin-1); Hubbard et al., Annal Int Med 3:206-212 (1989) (α1-antitrypsin); Smith et al., 1989, J Clin Invest 84:1145-1146 (α-1-proteinase); Oswein et al., 1990, "Aerosolization of Proteins," Proceedings of Symposium on Respiratory Drug Delivery II, Keystone, Colorado. Examples include March (recombinant human growth hormone); Debs et al., 1988, J Immunol 140:3482-3488 (interferon-gamma and tumor necrosis factor alpha); and Platz et al., U.S. Patent No. 5,284,656 (granulocyte colony-stimulating factor; incorporated herein by reference). Methods and compositions for pulmonary delivery of drugs for systemic effects are described in U.S. Patent No. 5,451,569 issued by Wong et al. on September 19, 1995 (its disclosure relating thereto is specifically incorporated herein by reference).

[0607] The use of a wide range of mechanical devices designed to deliver therapeutic products to the lungs, including but not limited to nebulizers, dose-measuring inhalers, and powder inhalers, all of which are well known to those skilled in the art, is contemplated.

[0608] Nasal delivery of pharmaceutical compositions is also being considered. Nasal delivery allows the pharmaceutical composition to enter the bloodstream directly after the product is administered through the nose, without requiring the accumulation of therapeutic products in the lungs. Formulations for nasal delivery include those containing dextran or cyclodextran.

[0609] When systemic delivery is desired, the compounds can be formulated for parenteral administration by injection, for example, by bolus injection or continuous infusion. Injectable formulations may be supplied in unit dosage forms, for example, in ampoules or in repeat-dose containers with added preservatives. Compositions may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles and may contain compounding agents such as suspending agents, stabilizers, and / or dispersants.

[0610] Pharmaceutical formulations for parenteral administration contain aqueous solutions of the active compound in a water-soluble form. Furthermore, suspensions of the active compound can be prepared as suitable oily suspensions for injection. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. Aqueous suspensions for injection may contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or activators that improve the solubility of the compound, allowing for the preparation of very high-concentration solutions.

[0611] Alternatively, the active compound may be in powder form for preparation using a suitable vehicle, such as a sterile pyrogen-free substance, before use.

[0612] The compounds can also be formulated into rectal or vaginal compositions, such as suppositories or enemas, which may contain conventional suppository bases, such as cocoa butter or other glycerides.

[0613] In addition to the formulations described above, the compounds can also be formulated as depot preparations. Such long-acting formulations can be formulated with suitable polymeric or hydrophobic substances (e.g., as emulsions in acceptable oils), or with ion exchange resins, or as somewhat poorly soluble derivatives, for example, as somewhat poorly soluble salts.

[0614] The pharmaceutical composition may also contain suitable solid-phase or gel-phase supports or excipients. Examples of such supports or excipients include, but are not limited to, polymers such as calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polyethylene glycol.

[0615] Suitable forms of the pharmaceutical preparations, whether liquid or solid, include, for example, aqueous or saline solutions for inhalation, microcapsule-encapsulated, encochleate, coated with fine gold particles, contained in liposomes, sprayed, aerosol, pellets for skin implantation, or dried on a sharp object for skin rubbing. The pharmaceutical compositions also include granules, powders, tablets, coated tablets, (micro)capsules, suppositories, syrups, emulsions, suspensions, creams, drops, or preparations with sustained release of the active compound, wherein excipients and additives and / or auxiliary agents such as disintegrants, binders, coatings, swelling agents, lubricants, flavorings, sweeteners, or solubilizers are conventionally used in the preparations described above. The pharmaceutical compositions are suitable for use in a variety of drug delivery systems. For a brief overview of drug delivery methods, see Langer R, Science 249:1527-1533 (1990).

[0616] Compounds and, if applicable, one or more other therapeutic agents may be administered either in themselves (undiluted) or in the form of pharmaceutically acceptable salts. While salts should be pharmaceutically acceptable when used in medicine, non-pharmaceutically acceptable salts may be conveniently used to prepare pharmaceutically acceptable salts of that salt. Such salts include, but are not limited to, those prepared from the following acids: hydrogen chloride, hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid, maleic acid, acetic acid, salicylic acid, p-toluenesulfonic acid, tartaric acid, citric acid, methanesulfonic acid, formic acid, malonic acid, succinic acid, naphthalene-2-sulfonic acid, and benzenesulfonic acid. Similarly, such salts may be prepared as alkali metal salts or alkaline earth salts, such as sodium, potassium, or calcium salts of the carboxylic acid group.

[0617] Suitable buffering agents include acetic acid and salt (1-2% w / v); citric acid and salt (1-3% w / v); boric acid and salt (0.5-2.5% w / v); and phosphoric acid and salt (0.8-2% w / v). Suitable preservatives include benzalkonium chloride (0.003-0.03% w / v); chlorobutanol (0.3-0.9% w / v); parabens (0.01-0.25% w / v); and thimerosal (0.004-0.02% w / v).

[0618] The pharmaceutical composition contains an effective amount of the compounds described herein and, optionally, one or more other therapeutic agents contained in a pharmaceutically acceptable carrier. The term “pharmaceutically acceptable carrier” means one or more suitable solid or liquid fillers, diluents or encapsulating materials suitable for administration to humans or other vertebrates. The term “carrier” refers to a natural or synthetic organic or inorganic component that facilitates the administration of the active ingredients together. The components of the pharmaceutical composition may also be mixed with and with the compounds in such a way that no interactions are likely to substantially impair the desired pharmaceutically effectiveness.

[0619] Specifically, therapeutic agents, including compounds, may be supplied in particles. “Particles,” as used herein, mean nanoparticles or microparticles (or, in some examples, larger particles) that may consist of all or part of the compounds or other therapeutic agents described herein. Particles may contain the therapeutic agent in a core surrounded by a coating agent, including, but not limited to, an enteric coating. The therapeutic agent may also be dispersed throughout the particles. The therapeutic agent may also be adsorbed onto the particles. Particles may have any order of release rate, including zero-order release, primary release, secondary release, delayed release, sustained release, immediate release, and any combination thereof. In addition to the therapeutic agent, particles may contain any of the substances commonly used in the fields of compounding and pharmaceuticals, including, but not limited to, edible substances, non-edible substances, biodegradable substances or non-biodegradable substances, or combinations thereof. Particles may be microcapsules containing compounds in solution or semi-solid state. Particles may have substantially any shape.

[0620] Both non-biodegradable and biodegradable polymer materials can be used in the manufacture of particles for delivering therapeutic agents. Such polymers may be natural or synthetic polymers. The polymers are selected based on the desired release period. Of particular interest are bioadhesive polymers, including biodegradable hydrogels described in Sawhney et al., Macromolecules 26:581-587 (1993), whose teachings are specifically incorporated herein by reference. These include polyhyaluronic acid, casein, gelatin, glutin, polyanhydride, polyacrylic acid, alginate, chitosan, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butyl methacrylate), poly(isobutyl methacrylate), poly(hexyl methacrylate), poly(isodecyl methacrylate), poly(lauryl methacrylate), poly(phenyl methacrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), and poly(octadecyl acrylate).

[0621] Therapeutic agents may be contained in controlled-release systems. The term “controlled-release” is intended to refer to any drug-containing formulation in which the mode and profile of drug release from the formulation are controlled. This includes immediate-release and non-immediate-release formulations, non-immediate-release formulations including sustained-release and delayed-release formulations, but not limited to these. The term “sustained-release” (also called “sustained-release”) is used in its idiomatic sense to refer to a drug formulation that achieves a gradual release of the drug over a long period, allowing for a substantially constant blood level of the drug over a long period. The term “delayed-release” is used in its idiomatic sense to refer to a drug formulation in which there is a delay time between the administration of the drug formulation and the subsequent release of the drug. “Delayed-release” may or may not include the gradual release of the drug over a long period, and therefore may or may not be “sustained-release.”

[0622] For the treatment of chronic conditions, the use of long-term sustained-release implants may be particularly suitable. “Long-term” release, as used herein, means that the implant is configured and prepared to deliver therapeutic levels of the active ingredient for at least 7 days, and up to 30–60 days. Long-term sustained-release implants are well known to those skilled in the art and include some of the release systems described above.

[0623] Treatment method Methods for treating inflammatory diseases or disorders are also provided. These methods may involve modulating the activity of activated fibroblasts. As used herein, the term “modulate” and its variations mean altering or inducing a modification of a particular biological activity. Modulation includes, but is not limited to, stimulating or inhibiting activity by, for example, activating a receptor to inhibit it from transmitting a signaling pathway, by activating an endogenous inhibitor that weakens biological activity, or by inhibiting the activity of a protein that inhibits a particular biological function.

[0624] The method may include a step of administering a compound (e.g., a conjugate) of any of the formulas presented herein. The method may also include a step of administering any of the pharmaceutical compositions described herein. The method may also include a step of administering a therapeutically effective amount of a compound (e.g., a conjugate) (whether as part of a pharmaceutical composition or otherwise) provided herein.

[0625] In some embodiments, inflammatory diseases or disorders are selected from the group consisting of 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 fibrous disease, graft-versus-host disease (GVHD), transplant rejection, fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, prosthesis lysis, glomerulonephritis, scleroderma, psoriasis with autoimmune myocarditis, spinal cord injury, central nervous system disorders, viral infections, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, and atherosclerosis.

[0626] A method for treating cancer is provided. The method for treating cancer may involve modulating the activity of activated fibroblasts. The method may include the step of administering a compound of any of the formulas provided herein (e.g., a conjugate) and / or a pharmaceutical composition provided herein. The method may include the step of contacting a CAF (e.g., a CAF of a cancer patient) with a compound of any of the formulas provided herein (e.g., a conjugate). In some embodiments, the cancer is lung cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, neck cancer, cutaneous melanoma, intraocular melanoma, uterine cancer, ovarian cancer, endometrial cancer, leiomyosarcoma, rectal cancer, gastric cancer, colon cancer, breast cancer, triple-negative breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, non-small cell lung cancer, small cell lung cancer. The cancers are selected from the group consisting of adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, pleural mesothelioma, bladder cancer, Burkitt lymphoma, ureteral cancer, kidney cancer, renal cell carcinoma, renal pelvis cancer, neoplasms of the central nervous system (CNS), primary CNS lymphoma, spinal axis tumor, brainstem glioma, pituitary adenoma, cholangiocarcinoma, Hurthle cell thyroid cancer, and gastroesophageal junction adenocarcinoma. In some embodiments, the cancers are selected from the group consisting of lung cancer, breast cancer, colorectal cancer, cervical cancer, and brain cancer (e.g., glioblastoma).

[0627] Methods for treating fibrosis are also provided. These methods may involve modulating the activity of activated fibroblasts. The method may include administering a compound of any of the formulas provided herein (e.g., a conjugate) (e.g., as part of a pharmaceutical composition provided herein, or otherwise) (e.g., in a therapeutically effective amount). In some embodiments, the fibrosis is selected from the group consisting of pulmonary fibrosis, renal fibrosis, and hepatic fibrosis. In some embodiments, the fibrosis is idiopathic pulmonary fibrosis.

[0628] A method for treating fibrosis or cancer may further include steps performed on a target of chemotherapy or radiotherapy. In certain embodiments, the method for treating fibrosis or cancer includes the step of administering a compound of any of the formulas provided herein (e.g., a conjugate) (e.g., as part of a pharmaceutical composition provided herein or otherwise) alone (e.g., in a therapeutically effective amount). In certain embodiments, the method for treating fibrosis or cancer includes the step of administering a compound of any of the formulas provided herein (e.g., a conjugate) (e.g., as part of a pharmaceutical composition provided herein or otherwise) (e.g., as part of a pharmaceutical composition provided herein or otherwise) to a target in combination with one or more additional therapies. Such additional therapies may, but are not limited to, include immunotherapy, DNA damage response pathway inhibitors, chemotherapy and / or surgery.

[0629] A method is provided for imaging cancer or fibrosis in a subject having cancer or fibrosis. In certain embodiments, the method includes administering to a subject an effective amount (e.g., a therapeutically effective amount) of any of the compounds provided herein (e.g., a conjugate) (e.g., as part of a pharmaceutical composition provided herein or otherwise). In certain embodiments, the method further includes imaging the subject. In certain embodiments, the method further includes generating an image of cancer or fibrosis in the subject (e.g., after or simultaneously with the administration of the compound).

[0630] Similarly, compositions and methods for optical imaging are provided. These compositions and methods may be used for fluorescence-guided surgery. These compositions and methods may be used for radiographic imaging.

[0631] This disclosure also relates to compositions and methods for magnetic resonance imaging (MRI).

[0632] The above method comprises the step of providing an effective amount of conjugate ALB to a patient in need thereof, wherein A comprises a FAPα target-directing moiety such as a moiety having a molecular weight of less than 10,000, L comprises one or more linkers capable of forming chemical bonds with at least A and B', and B' comprises an optical dye (e.g., a fluorescent dye), a photodynamic agent, a radioimaging agent, a radiotherapy agent, a chemotherapeutic agent, an antifibrotic agent, or an anticancer agent effective against cancer cells, cancer-associated fibroblasts, myofibroblasts and / or other tumor microenvironment factors.

[0633] The radiographic imaging agent may be a magnetic resonance (MR) contrast agent. The MR contrast agent may contain iron oxide particles. The iron oxide particles may be nanoparticles. The iron oxide particles may be paramagnetic particles, superparamagnetic particles (SPIOs), or ultra-small superparamagnetic particles (USPIOs).

[0634] Methods to improve ligand affinity of FAP Methods for improving the ligand affinity of FAP are also provided. In certain embodiments, a method for improving the ligand affinity of FAP (such ligands include isoindoline scaffolds) includes the step of introducing a triazole moiety to isoindoline or another scaffold of the ligand by molecular modeling to achieve a higher Schrödinger molecular docking score, thereby improving the ligand affinity of FAP. In certain embodiments, the method may include the step of introducing an ethyldiaminoaryltriazole moiety to the FAP ligand. In certain embodiments, the method may include the step of introducing a triazole moiety and a phenyl ring to the FAP ligand by molecular modeling to achieve a higher Schrödinger molecular docking score.

[0635] definition Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art in the fields of chemistry and biology. Furthermore, where used herein and in the appended claims, the singular forms "a," "an," and "the" refer to multiple subjects unless the content specifically indicates otherwise. Thus, for example, if a compound / composition is substituted with "one" alkyl or aryl, the compound / composition is optionally substituted with at least one alkyl and / or at least one aryl.

[0636] The terms “preventive or therapeutic” treatments are recognized in the art and include the administration of one or more of the compounds of this disclosure to a patient. The treatment is preventive (i.e., the treatment protects the host from the onset of the undesirable condition) if it is performed before the clinical manifestations of an undesirable condition (e.g., disease of the host animal or other undesirable condition), whereas the treatment is therapeutic (i.e., the treatment is intended to reduce, improve or stabilize an existing undesirable condition or its side effects) if it is performed after the signs of an undesirable condition.

[0637] The terms “patient,” “individual,” or “subject” refer to a mammal requiring a specific treatment. The patient or subject may be a primate, canid, feline, or equid. The patient or subject may be a bird. The bird may be a domesticated bird such as a chicken. The bird may be poultry. The patient or subject may be a human.

[0638] "Oxo" refers to the =O radical.

[0639] "Alkyl" refers to a group of 1 to 15 carbon atoms (for example, C1 to C1). 15The term "alkyl" generally refers to linear or branched hydrocarbon chain radicals consisting only of carbon and hydrogen atoms, such as those having an alkyl group. Unless otherwise specified, the disclosure of "alkyl" provided herein is intended to include a separate enumeration of saturated "alkyl" groups. Alkyl groups are defined as having 1 to 13 carbon atoms (e.g., C1 to C13). 13 Alkyl groups can contain 1 to 8 carbon atoms (e.g., C1-C8 alkyl groups). Alkyl groups can contain 1 to 5 carbon atoms (e.g., C1-C5 alkyl groups). Alkyl groups can contain 1 to 4 carbon atoms (e.g., C1-C4 alkyl groups). Alkyl groups can contain 1 to 3 carbon atoms (e.g., C1-C3 alkyl groups). Alkyl groups can contain 1 to 2 carbon atoms (e.g., C1-C2 alkyl groups). Alkyl groups can contain 1 carbon atom (e.g., C1 alkyl groups). Alkyl groups can contain 5 to 15 carbon atoms (e.g., C5-C 15 Alkyl groups may include alkyl groups. Alkyl groups may include 5 to 8 carbon atoms (e.g., C5 to C8 alkyl groups). Alkyl groups may include 2 to 5 carbon atoms (e.g., C2 to C5 alkyl groups). Alkyl groups may include 3 to 5 carbon atoms (e.g., C3 to C5 alkyl groups). In other embodiments, alkyl groups are selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). Alkyl groups are bonded to the rest of the molecule by single bonds.

[0640] "Alkoxy" refers to a radical in which the oxygen atom of the formula -O-alkyl is bonded, and alkyl is an alkyl chain as defined above.

[0641] "Alkylene" or "alkylene chain" generally refers to a straight or branched divalent alkyl group that has 1 to 12 carbon atoms and links the rest of the molecule to a radical group, such as methylene, ethylene, propylene, i-propylene, and n-butylene.

[0642] The term "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. An aromatic monocyclic or polycyclic hydrocarbon ring system contains only hydrogen and 5 to 18 carbon atoms, in which case at least one of the rings in the ring system is completely unsaturated; that is, it contains a delocalized (4n+2)π electron system of the ring, according to Huckel's theory. Ring systems from which the aryl group is derived include, but are not limited to, groups such as benzene, fluorene, indan, indene, tetralin, and naphthalene.

[0643] "Aralkyl" or "aryl-alkyl" refers to the formula -R c -The aryl radical, R c However, this refers to radicals, such as the alkylene chains defined above, for example, methylene and ethylene. The alkylene chain portion of the aralkyl radical is sometimes substituted with respect to the alkylene chain, as described above.

[0644] "Carbocyclyl" or "cycloalkyl" refers to a stable, non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, including condensed or bridging ring systems having 3 to 15 carbon atoms. Carbocyclyls can contain 3 to 10 carbon atoms. Carbocyclyls can contain 5 to 7 carbon atoms. Carbocyclyls are bonded to the rest of the molecule by single bonds. Carbocyclyls or cycloalkyls are either saturated (i.e., containing only CC single bonds) or unsaturated (i.e., containing one or more double or triple bonds). Examples of saturated cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Unsaturated carbocyclyls are also called "cycloalkenyls." Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norborneyl, dekalinyl, and 7,7-dimethyl-bicyclo[2.2.1]heptanyl.

[0645] "Carbocyclylalkyl" refers to the formula -R c -A radical of carbocyclyl, R c This refers to the radical, which is an alkylene chain as defined above.

[0646] "Chelating group" or "chelate group" as used herein refers to a polydentate chemical group that can bond to a central metal atom via multiple bonding interactions by using two or more bonding sites on the chelate group. The combination of a chelate group and a metal atom is a chelate. The bond between the chelate group and the metal atom can be by non-covalent interactions or bonds. In some embodiments, the bond of the chelate group to the metal atom is by multiple coordination bonds. Chelate groups include, but are not limited to, DOTA, NOTA, and EDTA.

[0647] "Metals suitable for radiation imaging, radiation therapy, or magnetic resonance imaging" or "isotopes suitable for radiation imaging, radiation therapy, or magnetic resonance imaging" are not limited to the following, 18 F, 32 P, 44 Sc, 47 Sc, 52 Mn, 55 Co, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 86 Y, 89 Sr, 89 Zr, 90 Y, 99m Tc, 111 In, 114m In, 117m Sn, 124 I, 125 I, 131 I, 149 Tb, 153 Sm, 152 Tb, 155 Tb, 161 Tb, 169 Er, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 224 Ra, 225 Ab, 225 and 227 Th is included. In some embodiments, metals (or isotopes) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging are 177 It is Lu. In some embodiments, a metal (or isotope) suitable for radiation imaging, radiotherapy, or magnetic resonance imaging is 111 It is In.

[0648] "Halo" or "halogen" refers to bromo, chloro, fluoro, or iodo substituents.

[0649] "Haloalkyl" refers to an alkyl radical as defined above, which is substituted with one or more halogen radicals as defined above, such as trifluoromethyl, difluoromethyl, fluoromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, etc.

[0650] The term "heteroalkyl" refers to an alkyl group as defined above, in which one or more of the alkyl backbone carbon atoms are substituted by heteroatoms (having an appropriate number of substituents or valencies; for example, -CH2- can be replaced by -NH- or -O-). For example, each substituted carbon atom is independently substituted by a heteroatom, such as the carbon being substituted by nitrogen, oxygen, selenium or other suitable heteroatoms. In some examples, each substituted carbon atom is independently substituted by oxygen, nitrogen (e.g., -NH-, -N(alkyl)- or -N(aryl)-, or another substituent as intended herein), or sulfur (e.g., -S-, -S(=O)- or -S(=O)2-). The heteroalkyl group is bonded to the rest of the molecule at the carbon atoms of the heteroalkyl group. The heteroalkyl group is bonded to the rest of the molecule at the heteroatoms of the heteroalkyl group. The heteroalkyl group is C1~C 18 It is a heteroalkyl group. Heteroalkyl groups are C1-C 12 It is a heteroalkyl. A heteroalkyl is a C1-C6 heteroalkyl. A heteroalkyl is a C1-C4 heteroalkyl. A heteroalkyl may include alkoxy, alkoxyalkyl, alkylamino, alkylaminoalkyl, aminoalkyl, heterocycloalkyl, heterocycloalkyl, and heterocycloalkylalkyl as defined herein.

[0651] "Heteroalkylene" refers to the divalent heteroalkyl group defined above, which links one part of a molecule to another part of a molecule.

[0652] "Heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring radical that may contain 2- to 12 carbon atoms and 1- to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically stated herein, heterocyclyl radicals are monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include aromatic, fused, and / or bridging ring systems. The heteroatoms in heterocyclyl radicals may be oxidized. Heterocyclyl radicals are partially or completely saturated. The disclosure of "heterocyclyl" provided herein is intended to include a separate enumeration of heterocyclyls, including aromatic and non-aromatic ring structures, unless otherwise specified. Heterocyclyls are bonded to the rest of the molecule via any atom of the ring. Examples of such heterocyclyl radicals include, but are not limited to, dioxolanil, thienyl[1,3]dithianil, decahydroisoquinolyl, imidazolinil, 1,3-benzodioxolyl, 1,4-benzodioxanil, tetrahydroquinolinil, 5,6,7,8-tetrahydroquinazolinil, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinil, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinil, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinil, and indolinil. This includes isoindolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianil, tetrahydropyranil, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0653] An "N-heterocyclyl" or "N-bonded heterocyclyl" refers to a heterocyclyl radical as defined above, containing at least one nitrogen atom, where the bond site of the heterocyclyl radical to the rest of the molecule is via the nitrogen atom in the heterocyclyl radical. Examples of such N-heterocyclyl radicals include, but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

[0654] "Heteroaryl" refers to a radical derived from a 3- to 18-membered aromatic ring radical, which may contain 2- to 17 carbon atoms, as well as 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 in which at least one of the rings in the ring system is completely unsaturated, i.e., it contains a delocalized (4n+2)π-electron system of the ring, according to Huckel theory. Heteroaryls include fused or bridging ring systems. Heteroatoms in a heteroaryl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. Heteroaryls are bonded to the rest of the molecule via any atom of the ring. Examples of heteroaryls include, but are not limited to, azepinyl, acridinyl, benzimidazolyl, benzoindolyl, benzofuranil, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, benzonaphthofuranil, benzoxazolyl, benzodioxolyl, benzodioxynyl, benzopyranil, benzopyranonil, benzofuranil, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyrimidinyl, carbazolyl, sinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclo Lopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]sinnolinyl, 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, indolidinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthilidinyl, 1,6-naphthilidinol, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxyranil, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxadinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyr This includes pridine, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridadinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyrimidinyl (pridinyl), and thiophenyl (i.e., thienyl).

[0655] The term “radical,” as used herein, refers to a fragment of a molecule that has a free valency for bond formation. A monovalent radical has one free valency so that it can form one bond with another chemical group. Unless otherwise specified, a radical of a molecule (e.g., a radical of an FAP ligand) is produced by removing one hydrogen atom from the molecule to produce a monovalent radical having one free valency at the position where the hydrogen atom was removed. Where appropriate, radicals may also be divalent, trivalent, etc., by removing two, three or more hydrogen atoms or other groups to produce radicals that can bond to two, three or more chemical groups. Where appropriate, the free valency of a radical can be produced by removing non-hydrogen atoms (e.g., halogens) or by removing two or more atoms (e.g., hydroxyl groups), provided that the atoms removed constitute a small proportion of the total atoms in the molecule forming the radical (less than 20% of the total number of atoms). In some embodiments, radicals are formed by removing a hydroxyl group from a folate ester, antifolate agent, or folate analog.

[0656] The term “releasing linker,” as used herein, refers to a linker containing at least one cleavable bond that can be cleaved under extracellular physiological conditions (e.g., a pH-unstable bond, an acid-unstable bond, an oxidatively unstable bond, or an enzyme-unstable bond). Releasing groups also include photochemically cleavable groups. Examples of photochemically cleavable groups include linkers containing the 2-(2-nitrophenyl)-ethane-2-ol group 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)). Cleavage bonds (one or more) can be present within a cleavage linker and / or at one or both ends of a cleavage linker. It should be recognized that such physiological conditions resulting in bond cleavage include, for example, standard chemical hydrolysis reactions occurring as a result of compartmentalization in organelles such as endosomes at physiological pH or at pH lower than cytoplasmic pH. Exemplarily, the divalent linkers described herein may undergo cleavage under other physiological or metabolic conditions, such as through the action of glutathione-mediated mechanisms. It is understood that the instability of cleavage bonds can be regulated by incorporating functional groups or fragments within the divalent linker L, also known as adjacent auxiliaries, which are capable of supporting or promoting such bond cleavage. The instability of the cleavage bond can also be regulated by substitutions at or near the cleavage bond, such as including an alpha branch adjacent to the cleavage disulfide bond, increasing the hydrophobicity of substituents on the silicon in the portion having a hydrolyzable silicon-oxygen bond, or homologating the alkoxy group that forms the hydrolyzable ketal or acetal portion.Furthermore, it is recognized that additional functional groups or fragments may be included within the divalent linker L, which can support or promote further fragmentation of the compound after the cleavage of the release linker (if present).

[0657] It will be understood by those skilled in the art that other suitable modifications and adaptations to the compositions and methods described herein are readily apparent from the descriptions contained herein in light of information known to those skilled in the art and can be made without departing from the scope of this disclosure or any of its embodiments.

[0658] All patents, patent application publications, academic articles, teaching materials, and other publications referenced herein represent the level of skill of a person skilled in the art to which this disclosure relates. All such publications are incorporated herein by reference to the same extent as they are specifically and individually indicated, as each of the individual publications is incorporated by reference.

[0659] Numerous specific details are provided above to ensure a full understanding of the disclosure. Certain examples can be implemented without some or all of these specific details, and it should be understood that the disclosure is not limited to specific biological systems, specific cancers, or specific organs or tissues. The disclosure may, of course, vary, but remains applicable given the data presented herein.

[0660] Furthermore, various techniques and mechanisms of this disclosure sometimes describe relationships or connections between two components. Words such as combined, related, linked, associated, and their inflectional morphemes and similar terms are used interchangeably unless otherwise specified or made clear from the context. These words and expressions do not necessarily indicate a direct relationship, but include relationships through mediating components. A relationship between two components does not necessarily mean a direct, unhindered relationship, since various other components may exist between the two components of interest. Thus, unless otherwise specified, a relationship does not necessarily mean a direct, unhindered relationship.

[0661] Furthermore, this disclosure is presented herein solely for illustrative purposes, and it should be understood that the principles and embodiments described herein may be applied to components of compounds and / or compositions having configurations other than those specifically described herein. In fact, it is explicitly intended that the components of the compositions and compounds of this disclosure may be modified to facilitate their desired applications.

[0662] Where ranges relating to physical properties such as molecular weight or chemical properties such as chemical formulas are used herein, it is intended that these ranges include all combinations and partial combinations of the scope of this specification and the particular embodiments.

[0663] Furthermore, when the term “about” refers to a number or numerical value or range (including, for example, integers, fractions, and percentages), it means that the number or numerical range referred to is an approximation within the range of laboratory variability (or within statistical experimental error), and therefore the numerical value or range may vary between 1% and 15% of the specified number or numerical range (for example, + / - 5% to 15% of the listed values), provided that a person skilled in the art will consider them equivalent to the listed values ​​(for example, having the same function or result). The term “comprises” (and related terms such as “comprise” or “comprises” or “having” or “including”) is not intended to exclude that in other particular embodiments, for example, embodiments such as any compound, composition of substance, composition, method or process described herein may “consist of” or “substantially consist of” the described features. The term "substantially" means that a value or range may allow a degree of variation, for example, within 90%, 95%, or 99% of the specified value or the specified boundary value of the range.

[0664] If a treatment method involves more than one procedure, compound, or composition, it will be understood that the order, timing, number, concentration, and amount of administration are limited only by the medical requirements and limitations of that procedure (i.e., two procedures may be administered to a subject, for example, simultaneously, sequentially, in order, alternatively, or in accordance with any other regimen).

[0665] Furthermore, in describing representative embodiments, this disclosure may present the method and / or process as a specific sequence of steps. The method or process should not be limited to a specific sequence of steps described herein to the extent that it does not rely on such a sequence of steps. As those skilled in the art will recognize, other sequences of steps are possible. Therefore, the specific sequence of steps disclosed herein should not be construed as a limitation on the claims. Moreover, the claims relating to the method and / or process should not be limited to performing those steps in the sequence described herein, and those skilled in the art will readily recognize that the sequence can vary and still remain within the spirit and scope of this disclosure. [Examples]

[0666] The following examples illustrate certain specific embodiments of the present disclosure and are not intended to limit the scope of the claimed invention.

[0667] [Example 1A] Synthesis of FAP5-carbamate-PI3K inhibitors (PI3Ki)

[0668] [ka]

[0669] Under an N2 atmosphere, 4-nitrophenyl chloroformate (20 mg, 0.1 mmol, 1.0 equivalent) was dissolved in dimethyl sulfoxide (DMSO) (1 mL). Next, diispropylethylamine (DIPEA) (15.48 mg, 0.12 mmol, 1.2 equivalents) and then compound 1 (58.85 mg, 0.11 mmol, 1.1 equivalents) were added. This mixture was maintained for 30 minutes. The reaction was monitored by liquid chromatography-mass spectrometry (LC-MS) until the starting materials were completely consumed, and then washed with ethyl acetate (EA) (2 mL) and water three times (3 × 2 mL). The product was dried over sodium sulfate and concentrated under reduced pressure, and purified by passing through a combi with Hex / EA as the eluent. Compound 2 was obtained as a yellow oily substance in 42 mg.

[0670] Under an N2 atmosphere, compound 2 (6.99 mg, 0.01 mmol, 1.0 equivalent) was dissolved in DMSO (1 mL), then FAP5 free amine (5.16 mg, 0.01 mmol, 1.0 equivalent) and then DIPEA (1.93 mg, 0.015 mmol, 1.5 equivalents) were added. This mixture was maintained for 2 hours. After purification, the final compound was obtained as a white powder in an amount of 1.4 mg. Purification conditions: reversed-phase C-18 column, ACN / NH4HCO3, pH=7, flow rate 8 mL / min.

[0671] [Example 1B] Comparison of various FAP5-PI3K inhibitors In 12-well plates, primary HLF cells at passage number 8 were seeded (approximately 100,000 cells / well) in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) to achieve approximately 80% confluence. The cells were then starved for 24 hours in DMEM containing 0.4% FBS, and subsequently stimulated for 24 hours with 10 ng / mL of transforming growth factor-beta-1 (TGF-β1).

[0672] After stimulation, cells were incubated with the FAP5-PI3K inhibitor for 2 hours, then replaced with culture medium containing 2 ng / mL of TGF-β1 and incubated for 24 hours (see Figure 21) and 48 hours (see Figure 22). 0.1% DMSO was added to all wells as a vehicle.

[0673] Cells were detached using trypsin, lysed in lysis buffer, and cell-derived proteins were extracted for Western blot analysis.

[0674] Figures 23A and 23B compare the inhibition of Akt phosphorylation by FAP5-PI3K inhibitors at 24 hours of incubation (Figure 23A) and 48 hours of incubation (Figure 23B). pAkt was normalized to tAkt. Positive controls were treated with TGF-β1 alone or untreated, while negative controls were treated without TGF-β1 or untreated. Among compounds with non-releasing linkers, FAP5-carbamate-PI3ki consistently performed better than FAP5-ester-PI3Ki and FAP5-PI3Ki-NR.

[0675] [Example 1C] Synthesis of FAP-3000

[0676] [ka]

[0677] Isoindoline-4-carboxylic acid 4-methyl hydrochloride was purchased from PharmaBlock (Hatfield, PA). Boc-L-pyroglutamate benzyl ester was purchased from Accela ChemBio (San Diego, CA). 4-(p-iodophenyl)butyric acid was purchased from AstaTech, Inc (Bristol, PA). NHS-ester-PEG6-NHFmoc and propargyl-PEG6-amine were purchased from BroadPharm. DOTA-NHS ester was purchased from Macrocyclics. Fmoc-Lys-OH was purchased from AAPPTec (Louisville, KY). 4,4-difluoro-L-prolineamide hydrochloride and HATU were purchased from Chem-Impex International (Chicago, IL). 4-ethynylbenzoic acid and mono-Fmoc ethylenediamine hydrochloride were purchased from AA Blocks LLC (San Diego, CA). Di-tert-butyl dicarbonate was purchased from Oakwood Chemical (Estill, SC). 10% carbon-supported palladium was purchased from Alfa Aesar (Haverhill, MA). Sodium borohydride, N-bromosuccinimide, triphenylphosphine, sodium azide, lithium bis(trimethylsilyl)amide, tert-butyl bromoacetate, 1,8-diazabicyclo[5.4.0]undeca-7-ene, pyridine, imidazole, phosphoryl chloride, diethyl ether (either), DIPEA, trifluoroacetic acid (TFA), tetrahydrofuran (THF), N,N-dimethylformamide (DMF), dichloromethane (DCM), methanol (MeOH), DMSO, and all other reagents were purchased from Sigma-Aldrich (St. Louis, MO). All synthesized molecules were purified using either flash chromatography (CombiFlash RF, Teledyne) or RP-HPLC (Agilent 1200 Instrument) with an XBridge OBD preparative column (19 × 150 mm, 5 μm) purchased from Waters (Milford, MA).Low-resolution mass spectrometry-liquid chromatography / mass spectrometry (LRMS-LC / MS) was performed using an Agilent 1220 Infinity LC equipped with a reversed-phase XBridge Shield RP18 column (3.0 × 50 mm, 3.5 μm).

[0678] The synthesis of key intermediate fragments 1 and 2 has been reported somewhere in the literature.

[0679] In short, 4-ethynylbenzoic acid was dissolved in anhydrous DMF containing HATU (1 equivalent) and anhydrous DIPEA (3 equivalents) for 10 minutes. Mono-Fmoc ethylenediamine was dissolved in anhydrous DMF and DIPEA (1.5 equivalents) and then added to the reaction mixture. The resulting solution was stirred under an inert atmosphere for 2 hours. The desired intermediate 1 was precipitated with ice-cold water, then filtered and dried under vacuum. Intermediate 1 was dissolved in anhydrous DMF, then CuI (0.5 equivalents) and DIPEA (2.0 equivalents) were added. This reaction mixture was heated to 55°C and stirred for 5 hours. The crude product was extracted with ethyl acetate (SiO), washed with ice-cold brine, and then purified by flash chromatography [A=Hex, B=SiO, solvent gradient from B0% to B100% over 20 minutes] to obtain product intermediate 2. LC / MS (m / z): [M+H]+C 40 H 40 The calculated mass of N6O5 is 684.8; the measured mass is 685.

[0680] Intermediate 2 was dissolved in DCM and diethylamine (20 equivalents), and then stirred for 1 hour. LC / MS (m / z): [M+H]+C 25 H 30The calculated mass of N6O3 was 462.6; the measured mass was 463. After deprotection was complete, the product was isolated by rotational evaporation, washed several times with ice-cold diethyl ether, and then placed under high vacuum for several hours. After this, it was used without further purification. NHS ester-PEG6-NHFmoc (1.1 equivalents) and DIPEA (3 equivalents) were added and stirred for several hours. The crude product was washed and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B30% over 25 minutes] to obtain intermediate 3. LC / MS (m / z): [M+H]+C 55 H 69 N6O 12 The calculated value is 1020.2; the measured mass is 1021.

[0681] Intermediate 3 was dissolved in ACN and cooled to 0°C. An equal volume of TFA was added, and the reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by LC / MS. LC / MS (m / z): [M+H] + C 50 H 61 N7O 10 Calculated value 920.1; measured mass 921. Once deprotection was complete, TFA was removed by rotary evaporation, and the deprotected product was used without further purification. Fragment 2 was dissolved in anhydrous DMF and DIPEA (3 equivalents) for 10 minutes. The previously deprotected product was dissolved in anhydrous DMF containing excess DIPEA, then added to the reaction mixture, and stirred under an inert atmosphere for 2 hours. The crude product was extracted with ethyl acetate, washed with ice-cold brine, and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B30% over 25 minutes] to obtain intermediate 4. [M+H] + C 62 H 72 F2N 10 O 13 Calculated value: 1203.3; measured value: 1204.

[0682] Intermediate 4 was dissolved in DCM and diethylamine (20 equivalents) and then stirred for 1 hour. LC / MS (m / z): Calculated value of [M+H]+C47H62F2N10O11 981.1; measured mass 982. After deprotection was complete, the product was isolated by rotational evaporation, washed several times with ice-cold diethyl ether, then placed under high vacuum for several hours, and used thereafter without further purification. The deprotected product was dissolved in anhydrous DMF and DIPEA (3 equivalents) containing DOTA-NHS ester (1.2 equivalents) and stirred under an inert atmosphere for 12 hours. The product was purified by RP-HPLC [A=20mM ammonium acetate buffer (pH 5.0) and B=CH3CN, solvent gradient from B5% to B55% over 45 minutes] to obtain the final desired product FAP-3000 (see Figure 24). [M+H]+C 63 H 88 F2N 14 O 18 Calculated value: 1367.47; measured value: 1368.

[0683] [Example 1D] Synthesis of FAP-3001

[0684] [ka]

[0685] 4-(p-iodophenyl)butyric acid was dissolved in anhydrous DMF containing HATU (1 equivalent) and anhydrous DIPEA (3 equivalents) for 1 hour. Fmoc-Lys-OtBu HCl (1 equivalent) was dissolved in anhydrous DMF and DIPEA (1.5 equivalents) and then added to the reaction mixture. The resulting solution was stirred under an inert atmosphere for 2 hours. The product was extracted with ethyl acetate, washed with ice-cold brine, and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B20% over 25 minutes] to obtain intermediate 5. LC / MS (m / z): [M+H] + C 31 H 33 The calculated mass of IN2O5 is 696.6; the measured mass is 697.

[0686] Intermediate 5 was dissolved in anhydrous DMF containing HATU (1 equivalent) and anhydrous DIPEA (3 equivalents) for 10 minutes. Propargyl-PEG6-amine was dissolved in anhydrous DMF and DIEPA (1.5 equivalents) and then added to the reaction mixture. The resulting solution was stirred under an inert atmosphere for 3 hours. The product was extracted with SiO2, washed with ice-cold brine, and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B20% over 25 minutes] to obtain intermediate 6. LC / MS (m / z): [M+H] + C 46 H 60 IN3O 10 The calculated value was 941.9; the measured mass was 943. The reaction progress was monitored by LC / MS.

[0687] Fragment 1 (1.2 equivalents) was dissolved in anhydrous DMF containing intermediate 6, and then CuI (0.5 equivalents) and DIPEA (2.0 equivalents) were added. The reaction mixture was heated to 55°C and stirred for 5 hours. The product was extracted with SiO2, washed with ice-cold brine, and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B20% over 25 minutes] to obtain intermediate 7. LC / MS (m / z): [M+H] + C 60 H 78 IN7O 12 Calculated value: 1216.2; measured value: 1217.

[0688] Intermediate 7 was dissolved in ACN and cooled to 0°C. An equal volume of TFA was added, and the reaction mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by LC / MS. LC / MS (m / z): [M+H] + C 55 H 70 IN7O 10The calculated value was 1116.11; the measured mass was 1117. After deprotection was complete, TFA was removed by rotary evaporation, and the product with the deprotected amine was used without further purification. Fragment 2 was dissolved in anhydrous DMF and DIPEA (3 equivalents) for 10 minutes. The previously deprotected product was dissolved in anhydrous DMF and excess DIPEA, then added to the reaction mixture and stirred under an inert atmosphere for 2 hours. The crude product was extracted with SiO2, washed with ice-cold brine, and then purified by flash chromatography [A=DCM, B=MeOH, solvent gradient from B0% to B30% over 25 minutes] to obtain intermediate 8. LC / MS (m / z): [M+H] + C 67 H 81 IN 10 O 13 Calculated value: 1399.3; measured value: 1400.

[0689] Intermediate 8 was dissolved in DCM and diethylamine (20 equivalents), and then stirred overnight. The reaction progress was monitored by LC / MS. LC / MS (m / z): [M+H] + C 52 H 71 F2IN 10 O 10 The calculated mass was 1177.11; the measured mass was 1178. After deprotection was complete, the amine-containing product was isolated by rotational evaporation, washed several times with ice-cold ethyl acetate, washed once with ice-cold ether, and then placed under high vacuum for several hours. After this, it was used without further purification. The deprotected product was dissolved in anhydrous DMF and DIPEA (3 equivalents) containing DOTA-NHS ester (1.2 equivalents) and stirred under an inert atmosphere for 12 hours. The final desired product FAP-3001 was purified by RP-HPLC [A=20 mM ammonium acetate buffer (pH 5.0) and B=CH3CN, solvent gradient from B5% to B55% over 45 minutes] to obtain FAP-3001 (see Figure 25). LC / MS (m / z): [M+H] + C 68 H 97 F2IN 14 O 18 Calculated value: 1563.6; measured value: 1564.

[0690] Other structures presented herein were synthesized in the same manner as the two compositions presented.

[0691] [Example 2] FAP conjugate binding affinity 250,000 HEK-FAP cells were seeded in amine-coated 24-well plates and allowed to reach confluence. Using the HEK-FAP cells, 10 nM FAP-rhodamine was replaced by increasing the concentration of the FAP-targeted conjugate at 4°C for 1 hour. The cells were then washed three times with phosphate-buffered saline (PBS), dissolved in 1% sodium lauryl sulfate (SDS), transferred to 96-well clear-bottom black-wall plates, and analyzed using a Synergy Neo2 microplate reader. All samples were triple-processed, and the bar for the standard error of the mean (SEM) is shown (see Figures 6A and 6B). 2.67 nM K d The inhibition constant was calculated using [the specified method].

[0692] [Example 3] Binding affinity of radiolabeled FAP conjugates Radiolabeling of the DOTA-containing conjugate was performed as follows: The conjugate was diluted in ammonium acetate (0.5 M, pH 8.0) to a final DOTA concentration of 0.5 mM. 111 In order to produce radiation imaging agents, 111 In( 111 With the addition of InCl3, a specific activity of up to 4.0 MBq / nmol was obtained, as shown, while the generation of the corresponding radiotherapy agent was as follows: 177 Lu( 177LuCl3 was added to obtain a specific activity of up to 11.0 MBq / nmol. Next, the resulting solution was heated to 90°C for 10–20 minutes, and the radiochemical purity of the labeled product was analyzed by radioisotope HPLC (20 mM ammonium acetate buffer (pH 7) (A) and acetonitrile (B)) with a linear gradient (B5% to B95% over 15 minutes). Radiochemical purity was found to be above 95% in all studies. After confirming the success of radiolabeling, diethylenetriaminepentaacetate sodium solution (5 mM, pH 7.0) was added to a final concentration of approximately 0.2 mM to complex any unreacted trace amounts of radioactive isotopes.

[0693] In the absence of, or in the presence of, 100x excess FAP competing ligands, 111 In-FAP-3000 or 111 Cancer-associated fibroblasts (Hs894 cell line) were incubated at room temperature for 1 hour while increasing the concentration of In-FAP-3001, washed three times with PBS, then dissolved in 1.0 M NaOH and analyzed by gamma counter (see Figures 4A and 4B). All samples were analyzed in a triple-bar SEM setup.

[0694] [Example 4] Consideration of drug administration for HT29 tumors Nu / nu mouse shoulder, 5x10 6 Human colorectal cancer cells with HT29 were inoculated. In nude mice with HT29 tumors, administration studies were conducted using either FAP-3000 or FAP-3001, radiolabeled with In-111, with the dose increased. MILabs VECTor 4+ Imaging was performed using instruments. Animals were anesthetized with isoflurane and scanned at various time points after injection. Emission scans were performed for 20–60 minutes using the MILabs VECTor / CT system. CT scans were acquired using an X-ray source set to 60kV and 615μA. U-SPECT II software and 171 111SPECT images were reconstructed in γ-energy width and at 241 keV. The POS-EM algorithm was used with 16 subsets and 4 iterations on a 0.8 mm voxel grid. CT images were reconstructed using NRecon software. The datasets were fused and filtered using PMOD software (version 3.2).

[0695] Nude mice with HT29 tumors were administered 0.3 mCi of In-111 chelated with either 20 nmol, 10 nmol, or 5 nmol of FAP-3000 via intravenous injection into the tail vein. Figure 5A illustrates mice that received FAP-3000 using single-photon emission tomography / computed tomography (SPECT / CT), with images taken 2 and 4 hours after injection.

[0696] Nude mice with HT29 tumors were administered 0.3 mCi of In-111 chelated with either 5 nmol or 1 nmol of FAP-3001 via intravenous injection into the tail vein. Figure 5B illustrates SPECT / CT images of such mice taken at various time points after injection (6 hours, 24 hours, 48 ​​hours, 72 hours, and 96 hours).

[0697] [Example 5] Consideration of administration for 4T1 tumors In the shoulders of Balb / cJ mice, 1 × 10⁶ units of sterile PBS were placed. 5 Individual 4T1 cells were inoculated. The administration study involved administering 0.3 mCi of In-111 chelated with either 30 nmol, 20 nmol, and 10 nmol of FAP-3000 or 6 nmol, 4 nmol, and 2 nmol of FAP-3001 to BALB / c mice with 4T1 tumors via intravenous injection into the tail vein. SPECT / CT images were taken at various intervals after injection. Figure 6A shows: 111 Figure 6B shows images from mice that received In-FAP-3000. 111The images shown are from mice that received In-FAP-3001.

[0698] [Example 6] Consideration of delay FAP-3001 retention studies were conducted in both BALB / c mice with 4T1 tumors and mice with KB tumors. Both sets of mice were injected with 5 nmol of FAP-3001 radiolabeled with 0.3 mCi of indium-111 via intravenous injection into the tail vein, and SPECT / CT images were taken at various post-injection intervals. Figure 7A shows images from BALB / c mice with 4T1 tumors, and Figure 7B shows images from mice with KB tumors.

[0699] [Example 7] The effect of albumin binders on the in vivo distribution of FAP In vivo distribution studies were conducted on 4T1 tumors with a volume of approximately 200 mm². 3 It started when it reached [a certain point]. The mice were given FAP-3000 radiolabeled with 100 μCi of lutetium-177. 177 Lu-FAP-3000), or 10 μCi of indium-111 ( 111 A single dose of 5 nmol of FAP-3001 (in-FAP-3001) was administered intravenously into the tail vein (see Figures 8A and 8B, respectively) and measured using a Packard Cobra gamma counter. Each desired time point contained 4-5 mice per conjugate, which had been induced into asphyxiation by CO2. The organs of interest were immediately collected, washed in cold PBS, and then measured using the gamma counter. Results were normalized to a percentage of the injected dose per gram of tissue.

[0700] [Example 8] In 4T1 tumors 177 Lu-FAP-3001 Radiation Therapy Review Treatment is for 4T1 tumors with a volume of approximately 50 mm. 3Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% ethanol (EtOH) in sterile PBS, while treatment mice were injected with FAP-3001 radiolabeled with either 0.25 mCi or 0.50 mCi of lutetium-177. 177 A single dose of 5 nmol of Lu-FAP-3001 was injected into the mice. Body weight of the mice was monitored as a measure of total toxicity. Figure 9A shows tumor growth data from this study, Figure 9B shows relative body weight data from this study, and Figure 9C shows a SPECT / CT scan of one of the treated mice 24 hours after injection.

[0701] [Example 9] In KB tumors 177 Lu-FAP-3000 Radiation Therapy Study Treatment is available for KB tumors with a volume of approximately 50 mm. 3 Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% EtOH in sterile PBS, while treated mice were injected with 0.50 mCi of lutetium-177 radiolabeled FAP-3000 ( 177 A single dose of 5 nmol of Lu-FAP-3000 was injected into the mice. The mice's body weight was monitored as a measure of overall toxicity. Figure 10A shows the tumor growth data from this study (KB tumor growth charts were used for nude mice for comparison), Figure 10B shows the survival curve from this study, and Figure 10C shows the relative body weight data of the mice derived from this study.

[0702] [Example 10] In KB tumors 177 Lu-FAP-3001 Radiation Therapy Review Treatment is available for KB tumors with a volume of approximately 50 mm. 3 Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% EtOH in sterile PBS, while treated mice were injected with 0.50 mCi of lutetium-177 radiolabeled FAP-3001 ( 177A single dose of 5 nmol of Lu-FAP-3001 was administered by injection. The mice's body weight was monitored as a measure of overall toxicity. Figure 11A shows the tumor growth data from this study (KB tumor growth charts were used for nude mice for comparison), Figure 11B illustrates the survival curve from this study, and Figure 11C shows the relative body weight data of the mice derived from this study.

[0703] [Example 11] In HT29 tumors 177 Lu-FAP-3001 Radiation Therapy Review Treatment is for HT29 tumors with a volume of approximately 150 mm. 3 Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% EtOH in sterile PBS, while treated mice were injected with 0.25 mCi of lutetium-177 radiolabeled FAP-3001 ( 177 A single dose of 5 nmol of Lu-FAP-3001 was injected into the mice. The mice's body weight was monitored as a measure of overall toxicity. Figure 12A shows the tumor growth data from this study (KB tumor growth charts were used for nude mice for comparison), Figure 12B illustrates the survival curve from this study, and Figure 12C shows the relative body weight data of the mice derived from this study. Figure 12D shows photographic images of the tumors dissected after euthanasia.

[0704] [Example 12] In U87MG tumors 177 Lu-FAP-3001 Radiation Therapy Review Treatment is for U87MG tumors with a volume of approximately 190 mm². 3 Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% EtOH in sterile PBS, while treated mice were injected with 0.50 mCi of lutetium-177 radiolabeled FAP-3001 ( 177A single dose of 5 nmol of Lu-FAP-3001 was administered by injection. The mice's body weight was monitored as a measure of overall toxicity. Figure 13A shows the tumor growth data from this study (KB tumor growth charts were used for nude mice for comparison), Figure 13B illustrates the survival curve from this study, and Figure 13C shows the relative body weight data of the mice derived from this study.

[0705] [Example 13] 177 Lu-FAP-3001 toxicological staining for radiation therapy To perform further toxicological evaluations, mice were randomly selected from a control group and a treatment group (0.5 mCi dose). The organs of interest were collected immediately after euthanasia, washed, and fixed in a 10% formalin buffer solution for 48–72 hours. The organs were then maintained in a 70% ethanol solution until radioactivity had completely decayed. After this, the organs were submitted to the histology laboratory at Purdue University for embedding in paraffin, sectioned, and stained with hematoxylin and eosin (H&E) (see Figure 14). Lesions in the tissue sections (n=4–8 per organ per mouse) were examined blindly by a board-certified pathologist at the Department of Comparative Pathobiology, Purdue University.

[0706] Table 1 summarizes the data observed from this study (see also Figure 14).

[0707] [Table 1]

[0708] [Example 14] 4T1 - High dose 177 Lu-FAP-3001 Radiation Therapy Review Treatment is for 4T1 tumors with a volume of approximately 200 mm². 3Treatment was initiated upon reaching [a certain point]. By intravenous injection into the tail vein, control mice were injected with 5% EtOH in sterile PBS, while treated mice received 1.50 mCi of lutetium-177 radiolabeled FAP-3001 ( 177 Lu-FAP-3001) 5 nmol as one dose, or on day 0 and day 3, FAP-3001 radiolabeled with 1.50 + 0.6 mCi of lutetium-177. 177 Two doses of Lu-FAP-3001)5 nmol were injected. The mice's body weight was monitored as a measure of total toxicity. Figure 15A shows tumor growth data from this study, Figure 15B shows the survival curve from this study, Figure 15C shows the relative body weight data of the mice derived from this study, and Figure 15D shows... 177 This image shows a SPECT / CT scan of one mouse that received a single dose of Lu-FAP-3001.

[0709] [Example 15] 4T1 - Blinded Measurement 177 Lu-FAP-3001 Radiation Therapy Review Treatment is for 4T1 tumors with a volume of approximately 100 mm². 3 Treatment began upon reaching [a certain point]. On day 0, control mice were injected with 5% EtOH in sterile PBS via intravenous injection into the tail vein, while treatment mice were injected with 1.50 mCi of lutetium-177 radiolabeled FAP-3001 ( 177 A single dose of 5 nmol of Lu-FAP-3001 was injected. Two assistants measured tumors with calipers every other day in a blinded manner (i.e., they did not know which mice were treated and which were controls). Mouse body weight was monitored as a measure of overall toxicity. Figure 16A shows tumor growth data from Study 7, Figure 16B shows the survival curve from this study, and Figure 16C shows the relative body weight data of mice derived from this study.

[0710] [Example 16] 4T1 - Blinded Measurement 177 Lu-FAP-3001 177 Investigation of radiotherapy using Lu-FAP-3005 Treatment is for 4T1 tumors with a volume of approximately 100 mm². 3 Treatment began upon reaching [a certain point]. On day 0, control mice were injected with 5% EtOH in sterile PBS via intravenous injection into the tail vein, while treatment mice were injected with FAP-3001 or FAP-3005 radiolabeled with 1.50 mCi of lutetium-177 (respectively, 177 Lu-FAP-3001 or 177 A single dose of 5 nmol of Lu-FAP-3005 was injected. Two assistants measured tumors with calipers every other day in a blinded manner (i.e., they did not know which mice were treated and which were controls). Mouse body weight was monitored as a measure of overall toxicity. Figure 17A shows tumor growth data from Study 7, Figure 17B shows the survival curve from this study, and Figure 17C shows the relative body weight data of mice derived from this study.

[0711] [Example 17] 177 Toxicological staining of Lu-FAP-3001 in high-dose radiotherapy To perform further toxicological evaluations, mice were randomly selected from control and treated mice (1.5 mCi dose) at various time points after injection. The organs of interest were collected immediately after euthanasia, washed, and fixed in a 10% formalin buffer solution for 48–72 hours. The organs were then maintained in a 70% ethanol solution until radioactivity had completely decayed. After this, the organs were submitted to the histology laboratory at Purdue University for embedding in paraffin, sectioned, and stained with H&E (see Figure 14). Lesions in the tissue sections (n=1–4 per organ per mouse) were examined blindly by a board-certified pathologist at the Department of Comparative Pathobiology, Purdue University. The table summarizes the results, with representative tissue sections shown in Figures 18A and 18B, respectively.

[0712] [Example 18] In a mouse pulmonary fibrosis model, disease was induced in C57 / BL6 mice by intratracheal injection of bleomycin, and on day 14,111 In-FAP-3001 (5 nmol, radiolabeled with 0.3 mCi) was administered intravenously, and a SPECT / CT scan was performed 24 hours later. See Figure 19.

[0713] [Example 19] High affinity FAP ligand and molecular docking The design of drug conjugates requires the specific selection of linker binding sites, ensuring that the binding affinity of the modified drug remains similar to that of the parent drug. Molecular docking experiments were used to identify suitable linker binding sites in known FAP inhibitors 1.

[0714] [ka]

[0715] The crystal structure of human FAP [PDB ID: 1Z68] was retrieved from the Protein Databank (a global open-access digital data source) and further prepared for docking using the protein preparation toolbox included in the Schrodinger software package (Schrodinger, LLC, New York, New York). The protein structure was preprocessed using Schrodinger's default protocol, and heteroatom states were generated using the Epik module with a pH value of 7.4+ / -0.5. The protein structure was further refined to optimize intramolecular hydrogen bonding and suppressed energy minimization using the OPLS4 force field.

[0716] The structures of three FAP ligands (FAP inhibitor 1, intermediate 2', and FAP-4000 (see structures of intermediate 2' and FAP-4000 below)) were uploaded to Maestro (Schrodinger, LLC, New York, NY), and preparations for grid docking were made using the LigPrep program (Schrodinger, LLC, New York, NY). The three-dimensional shapes of the ligands were optimized using the OPLS4 force field and used for docking.

[0717] The target ligand was docked to the FAP binding pocket using the standard Inductive Fit Docking (IFD) protocol of the Schrodinger software package. First, a receptor grid box was generated by identifying amino acid residues in FAP reported to be involved in the binding interaction. The IFD protocol, utilizing the grid docking protocol, generated up to 20 poses for each ligand, which were further refined using the Prime Refinement module. Residues within 5 Å of the ligand poses were refined, and the residue side chains were optimized. After refining the binding site following the initial docking, the ligand re-docked to a structure within 30.0 kcal / mol of the best structure and within the top 20 structures overall. A standard-precision model was used for the grid re-docking step.

[0718] Referring to Figure 27, it was found that in the docking pose between FAP inhibitor 1 and FAP, the Glu204 and Tyr541 residues are involved in non-covalent interactions. Glu204 forms a hydrogen bond with the NH group of the pyrrolidine ring, and Tyr541 forms a π-π interaction with the 2,3-dihydroisoindole ring of inhibitor 1. The shaded region of the 2,3-dihydroisoindole ring indicates the atom exposed to the solvent (Figure 27).

[0719] Next, FAP inhibitor 1 was further functionalized at the C-4 position of the 2,3-dihydroisoindole ring. Methyleneamine (-CH2NH2) was bonded at the C-4 position of the 2,3-dihydroisoindole ring of FAP inhibitor 1, forming intermediate 2'. When intermediate 2' was docked to the FAP protein, its interaction with the FAP protein was similar to that of parent FAP inhibitor 1, which has a further cation-pi interaction between the 2,3-dihydroisoindole ring and Arg550. The methyleneamine (-CH2NH2) in intermediate 2' remained exposed to the solvent.

[0720] In the deep binding pocket of FAP, the presence of nearby aromatic amino acid residues was observed, so we searched for hydrophobic spacers that would promote the formation of new, stable interactions with these aromatic amino acid residues. Thus, the methyleneamine group in intermediate 2' was replaced by a triazole ring containing a solvent-exposed methyleneamine (-CH2NH2) group (FAP-4000), resulting in a pi-pi interaction between the triazole ring and Phe350. Furthermore, the NH of methylamine formed a hydrogen bond with Cys545, and Arg123 formed a hydrogen bond with the amide oxygen adjacent to the 2,3-dihydroisoindole ring (see Figure 27). The increase in the number of intermolecular interactions improved the docking score from -8.0 kcal / mol (FAP inhibitor 1) to -9.4 kcal / mol (FAP-4000).

[0721] Since it was found that the C-1 position (-CH2NH2 unit) of the triazole ring of FAP-4000 was exposed to the solvent during docking, the methyleneamine in FAP-4000 was replaced with aminoethylbenzamide to form FAP-4001. When FAP-4001 was docked to the FAP protein, the docking score further improved to -11.5 kcal / mol. Specifically, the phenyl ring in FAP4001 formed a π-π interaction with Trp623, and the 2,3-dihydroisoindole ring formed a further cation-π interaction with Arg550.

[0722] Next, the modified structure was docked again into the same binding pocket (FAP4002) by utilizing the -NH2 portion of FAP-4001 for binding to the PEG4-amine linker and creating a larger grid box for accommodating the ligand-PEG4-amine drug conjugate. The interactions between the ligand and the Glu204 and Tyr541 residues were conserved in the docking pose between FAP4002 and FAP, resulting in a docking score of -9.5 kcal / mol. The triazole ring was involved in a pi-pi interaction with Phe350, as shown in Figure 27.

[0723] Therefore, these data support the idea that adding spacers and linkers to the parent (free) drug helps the drug maintain its binding affinity, even after conjugation, without interfering with the parent (free) drug's important non-covalent interactions.

[0724] [Example 20] Synthesis of FAP-Targeted Dye Compounds The free amine functional group in FAP-4002 was used to synthesize the FAP-targeted dye conjugate (FAP-4003) in accordance with Scheme 1 below:

[0725] [ka]

[0726] Next, as shown in Scheme 2, the synthesis of the FAP-targeted fluorescent compound was initiated from intermediates 5', 6', and 7'.

[0727] [ka] Option 1

[0728] [ka] Alternative option 2:

[0729] [ka]

[0730] In short, intermediate 5' was synthesized from commercially available benzyl pyroglutamate 8. Intermediate 6' was prepared from 2,3-dihydro-1H-isoindole-4-carbocite methyl hydrochloride 9 by tert-butyloxycarbonyl (Boc) protection in dichloromethane, yielding compound 10 in quantitative yield. Next, the methyl ester in compound 10 was reduced with NaBH4 by heating in methanol and tetrahydrofuran to obtain the corresponding alcohol 11 in 90% yield. Bromination of 11 with NBS / PPh3 in DMF at room temperature, followed by nucleophilic substitution of the corresponding bromide 12 with NaN3 in DMF at 70°C for 12 hours to obtain the key azide intermediate 6' in 95% yield. In dry DMF, in the presence of HATU / DIPEA, mono-Fmoc ethylenediamine hydrochloride was coupled with 4-ethynylbenzoic acid 13 to obtain alkyne intermediate 7'. Next, in DMF, at 55°C for 5 hours, intermediate 6' and intermediate 7' were subjected to a classical Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction to obtain click product 14 having a phenyltriazole side chain (appendage). Compound 14 was treated with trifluoroacetic acid (TFA) in DCM, then activated acid 5 was added, and then Fmoc was deprotected using diethylamine in DCM to obtain FAP-4001 (see Scheme 2, Option 1).

[0731] The same compound 14 was treated with diethylamine in dichloromethane for 30 minutes, and the resulting amine was then coupled with Fmoc-PEG4-NHS ester in CH2Cl2 in the presence of DIPEA to produce intermediate 15 in 90% yield (see Scheme 2, Option 2). Treatment of intermediate 15' in CH2Cl2 with TFA, followed by coupling of the resulting amine with a key acid intermediate 5' in DMF in the presence of HATU / DIPEA, yielded the FAP-targeting ligand (intermediate 16) in 65% yield.

[0732] Referring to Scheme 3, to produce the final FAP-targeted fluorescent dye used below, the Fmoc protecting group in compound 16 was deprotected with diethylamine in dichloromethane to produce free amine 17 (compound 17). FAP-4002 was obtained by coupling free amine 17 (compound 17) with 3-(4-hydroxyphenyl)propionic acid. Subsequently, under basic conditions, the FAP-targeted near-infrared dye FAP-4003 was obtained in 65% yield by substitution of chloride from the near-infrared dye (ClS0456).

[0733] [ka]

[0734] [Example 21] Internalization through compound bonding Regarding affinity for binding to FAP, the following compounds were analyzed: FAP inhibitor 1; FAP-targeted ligand with a linker (FTL-PEG4-NH2) (compound 17); conjugate of compound 17 with 4-hydroxyphenylpropionic acid (FAP-4002); conjugate of compound 17 with tetramethylrhodamine (FAP-4004); and conjugate of FAP-4002 with ClS0456 (FAP-4003).

[0735] [ka]

[0736] HT1080-FAP cells were incubated at 37°C for 1 hour with 25 nM FAP-4004 (see Figure 32A) or FAP-4003 (see Figure 32B), and examined by confocal microscopy and wide-field Nikon microscopy, respectively. The binding affinity and specificity of each compound were also quantified by measuring the fluorescence of either HT1080-FAP cells or HT1080 cells after incubation at 4°C for 1 hour in the presence or absence of 100-fold excess unlabeled FAP-4002, and in the presence of elevated concentrations of FAP-4004 (see Figure 32C) or FAP-4003 (see Figure 32D).

[0737] The ability of FAP-targeted FAP-4002 to inhibit the closely related dipeptidyl peptidases FAP, PREP, and DPP-IV was also evaluated. FAP, PREP, and DPP-IV were incubated with FAP-targeted FAP-4002 at room temperature for 10 minutes, after which a fluorescent substrate was added. The reaction was allowed to proceed for 30 minutes, after which it was stopped, and the change in fluorescence was quantified as a measure of catalytic activity. All assays were performed in triplicate, and SEM bars are shown (see Figures 33A-33C).

[0738] Synthesis example: The following examples further describe in detail the substances and methods used to synthesize the intermediates and compounds described in Examples 21-23 and Schemes 1-3. After synthesis, all intermediates and compounds / products were identified by LC-MS and nuclear magnetic resonance spectroscopy (NMR).

[0739] [Example 22] Synthesis of 2-(tert-butyl)4-methylisoindoline-2,4-dicarboxylate (compound 10) At room temperature, 20 mL of DCM was added to a stirred solution of 2,3-dihydro-1H-isoindole-4-carboxylate methyl hydrochloride 9 (1.00 g, 5.64 mmol). To this mixture, (Boc)2O (4.9 mL, 22.59 mmol) was added in one addition, followed by the addition of triethylamine (2.9 mL, 22.59 mmol) dropwise. The mixture was stirred for 12 hours, and then the reaction mixture was diluted with water (30 mL) and extracted into DCM (2 × 25 mL).

[0740] The organic layer was dehydrated with anhydrous MgSO4, filtered, and the filtrate was evaporated under reduced pressure. The resulting crude residue was purified by CombiFlash using hexane + ethyl acetate as the mobile phase, and compound 10 (1.4 g, 92%) was obtained as a white solid. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.92 (dd, J1= 15.0 Hz, J2= 10.0 Hz, 1H); 7.44-7.32 (m, 1H); 7.33 (m, 1H); 4.94 (d, J = 12.6 Hz, 2H); 4.67 (d, J = 17.0 Hz, 2H); 3.90 (s, 3H); 1.51 (s, 9H) ppm. 13 C-NMR (CDCl3, 125 MHZ) δ = 166.41, 154.50, 154.33, 139.93, 138.96, 138.83, 138.56, 129.37, 128.96, 127.61, 127.17, 126.89, LCMS of 4: LC / MS (m / z): C 15 H 21 Calculated value of NO4[M+H]: 278.14, measured value: 278.13 g / mol.

[0741] [Example 23] Synthesis of tert-butyl 4-(hydroxymethyl)isoindoline-2-carboxylate (alcohol 11) Under an N2 atmosphere, sodium bromide (1.37 g, 36.101 mmol) was added at room temperature to a stirred solution of compound 10 (1.0 g, 3.61 mmol) in THF (10.0 mL). Then, methanol (10 mL) was slowly added to the reaction mixture over 5 minutes. The reaction mixture was heated to 55°C and continuously stirred for 5 hours.

[0742] Next, the reaction mixture was cooled to 0°C, slowly quenched with saturated aqueous ammonium chloride, and extracted to dimethylammonium chloride (60 mL). The organic phase was collected, dehydrated with sodium sulfate, and the solvent was distilled to obtain a crude residue. This residue was purified using CombiFlash, yielding alcohol 11 (700 mg, 70%) as a viscous white solid. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.28-7.25 (m, 2H); 7.19-7.13 (m, 1H); 4.68-4.62 (m, 6H); 1.51 (s, 9H) ppm. 13 C-NMR (CDCl3, 125 MHZ) δ = 154.64, 137.60, 137.39, 135.62, 135.32, 143.92, 127.81, 125.90, 125.67, 122.05, 121.60, 79.87, LC-MS of 11. LC-MS (m / z): C 14 H 20 Calculated value of NO3[M+H]: 250.14, measured value: 250.14 g / mol.

[0743] [Example 24] Synthesis of tert-butyl 4-(bromomethyl)isoindoline-2-carboxylate (bromide 12) To a stirred solution of compound 11 (500 mg, 2.00 mmol) in DMF (10 mL), PPh3 (790 mg, 3.01 mmol) was added, followed by freshly recrystallized N-bromosuccinimide (NBS) (532 mg, 3.01 mmol). The reaction mixture was stirred under an N2 atmosphere at room temperature for 4-5 hours, then diluted with water (40 mL) and extracted with ethyl acetate (2 × 25 mL). The organic layer was washed with water and brine, then dehydrated with anhydrous sodium sulfate and filtered. The filtrate was evaporated under reduced pressure to obtain a crude residue, which was purified by CombiFlash to yield bromide 12 (450 mg, 90%) as a white solid. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.28-7.17 (m, 3H); 4.71 (m, 4H); 4.42 (d, J = 8.5 Hz, 2H); 1.52 (s, 9H) ppm. 13 C-NMR (CDCl3, 125 MHZ) δ = 154.46, 138.38, 138.03, 136.78, 136.48, 132.44, 132.17, 128.20, 128.13, 127.8, 123.18, 122.90, LC-MS of 12: LC-MS: (m / z): C 14 H 19 Calculated value of BrNO2[M+H]: 312.05, measured value: 312.05 g / mol.

[0744] [Example 25] Synthesis of tert-butyl 4-(azidomethyl)isoindoline-2-carboxylate (intermediate 6') To a stirred solution of bromide compound 12 (400 mg, 1.286 mmol) in DMF, NaN3 (420 mg, 6.430 mmol) was added, and the mixture was continuously stirred at 75°C for 6 hours. Next, the reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with water, brined, dehydrated with anhydrous sodium sulfate, and filtered. The filtrate was evaporated under reduced pressure and purified by CombiFlash, yielding azide intermediate 6' (300 mg, 95%) as a white solid. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.32-7.25 (m, 2H); 7.22-7.20 (m, 1H); 4.72-4.66 (m, 4H); 4.31 (s, 2H); 1.53 (s, 9H) ppm. 13 C-NMR (CDCl3, 125 MHZ) δ = 154.42, 138.28, 137.95, 136.34, 135.95, 130.18, 129.92, 128.08, 127.40, 127.30, 122.96, 122.65, LC-MS of 79.96, 52.59, 52.48, 52.39, 52.15, 51.05, 50.89, 28.56 ppm. 7: LC-MS (m / z): C 14 H 19 Calculated value of N4O2[M+H]: 275.14, measured value: 275.14 g / mol.

[0745] [Example 26] Synthesis of (9H-fluoren-9-yl)methyl(2-(4-ethynylbenzamide)ethyl)carbamate (intermediate 7') Under an N2 atmosphere, a stirred solution of 4-ethynylbenzoic acid compound 13 (500 mg, 3.424 mmol) in dry DMF (10 mL) was to which HATU (1.4 gm, 3.76 mmol) and then DIPEA (1.7 mL, 10.27 mmol) were added at room temperature. The mixture was continuously stirred for 10 minutes to activate the acid.

[0746] Next, N-Fmoc-ethylenediamine (1.0 g, 3.76 mmol) was added to the reaction mixture, and the mixture was continuously stirred for a further 3 hours. After this, the reaction mixture was diluted with water (50 mL), and the resulting precipitate was filtered through a Buchner funnel, yielding a white solid. This white solid was washed again with water (2 × 50 mL) and dried under vacuum for 1 hour to obtain intermediate 7' (1.2 g m, 85%). 1 7.25 (m, 2H); 4.44 (bs, 2H); 4.33 (d, J = 7.0 Hz, 1H); 4.16 (t, J = 7.0 Hz, 1H); 3.50 (s, 1H); 3.48 (t, J = 6.1Hz, 2H); 3.34(t, J = 6.0 Hz, 2H) ppm. 13 ¹¹C-NMR (CD3OD +CDCl3, 125 MHz) δ = 168.27, 158.16, 143.81, 141.20, 134.02, 131.83, 127.49, 127.06, 126.85, 125.62, 124.83, 119.65, 83.34, 79.69, 77.92, 66.61, 47.06, 40.04 ppm. and LC-MS of 7: LC-MS (m / z): ¹¹C 26 H 23 Calculated value of N2O3[M+H]: 411.16, measured value: 411.16 g / mol. HRMS-ESI: C 26 H 23 Calculated value of N2O3[M + H]+: 411.1708, measured value: 411.1710.

[0747] [Example 27] Synthesis of tert-butyl 4-((4-(4-((2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethyl)carbamoyl)phenyl)-1H-1,2,3-triazole-1-yl)methyl)isoindoline-2-carboxylate (Compound 14) To a mixture of azide intermediate 6 (1.0 equivalent) and alkyne intermediate 7 (1.2 equivalents) in dry DMF (5.0 mL), CuI (0.5 equivalents) was added, followed by DIPEA (2.0 equivalents). The reaction mixture was stirred under a nitrogen atmosphere at 55°C for 1 hour, then transferred to room temperature, diluted with saturated aqueous ammonium chloride (20 mL), and vigorously stirred for 15 minutes. The solid residue formed in the reaction mixture was filtered and washed with water (2 × 20 mL), and dried under vacuum for 1 hour to obtain compound 14 as a brown solid.

[0748] Compound 14 was used in some cases without purification. When a certain amount of compound 14 was purified using an EtOAC / hexane mixture as the mobile phase, compound 14 (97%) was obtained as a white solid. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.79 (bs, 3H); 7.78-7.70 (m, 2H); 7.68 (d, J = 7.9 Hz, 2H); 7.52 (d, J = 7.3 Hz, 2H); 7.73-7.720 (m, 7H); 7.15-7.13 (m, 1H); 5.50 (s, 2H); 4.67-4.57 (m, 4H); 4.34 (d, J = 6.7 Hz, 2H); 4.12 (t, J = 6.7 Hz, 1H); 3.48 (t, J = 5.7 Hz, 1H); 3.35-3.33 (m, 2H); 2.77 (s, 2H); 2.71 (bs, 1H); 1.46 (s, 9H) ppm. 13C-NMR (CDCl3, 125 MHZ) δ = 173.47, 167.26, 156.65, 154.33, 144.98, 141.27, 138.65, 138.46, 136.59, 135.92, 129.26, 129.03, 128.58, 127.82, 127.67, 127.44, 127.06, 125.61, 125.12, 123.58, 123.26, 119.95, 80.18, 80.08, 70.49, 70.13, 67.14, 66.58, 50.98, LC-MS of 14: LC-MS (m / z): C 40 H 41 Calculated value of N6O5[M+H]: 685.31, measured value: 685.31 g / mol. HRMS-ESI: C 40 H 41 Calculated value of N6O5[M + H]+: 685.3138, measured value: 685.3139.

[0749] [Example 28] Synthesis of tert-butyl 4-((4-(4-((1-(9H-fluoren-9-yl)-3,19-dioxo-2,7,10,13,16-pentaoxa-4,20-diazasodocosan-22-yl)carbamoyl)phenyl)-1H-1,2,3-triazole-1-yl)methyl)isoindoline-2-carboxylate (intermediate 15') To a stirred solution of compound 14 (400 mg, 0.584 mmol) in a DCM+MeOH mixture (1 + 0.5 mL), (Et)2NH (1.0 mL) was added and the mixture was continuously stirred for 2 hours. The reaction mixture was evaporated under reduced pressure, and the resulting crude residue was purified by CombiFlash using MeOH+CH2Cl2 as the mobile phase to obtain the free amine of compound 14 (this was reserved and used in the studies described herein).

[0750] A portion of the amine compound was dissolved in DCM (1 mL per 1 mmol), and Fmoc-NH(PEG)4NHS ester (1.2 equivalents) and DIPEA (2.0 equivalents) were added to this mixture under a nitrogen atmosphere at room temperature for 1 hour with stirring. The reaction mixture was evaporated under reduced pressure, and the resulting crude residue was purified using DCM + MeOH by CombiFlash, yielding intermediate 15' as a white solid with an 80% concentration. 1 H-NMR (CDCl3, 500 MHZ) δ = 7.86 (m, 4H); 7.74 (d, J = 7.5 Hz, 2H); 7.68 (d, J = 3.7 Hz, 1H); 7.57 (m, 3H); 7.37 (t, J = 7.5 Hz, 2H); 7.28 (m, 4H); 7.16 (m, 2H); 5.67 (bs, 1H); 5.51 (d, J=6.7 Hz, 2H); 4.71 (s, 2H); 4.65 (m, 2H); 4.37 (d, J=7.2Hz, 1H); Hz, 1H); 3.72-3.34 (m, 22H); 2.44 (s, 2H); 1.51 (s, 9H) ppm. 13 C-NMR (CDCl3, 125 MHZ) δ = 173.47, 167.26, 156.65, 154.33, 143.98, 141.27, 138.65, 138.46, 136.59, 135.92, 133.77, 133.19, 129.26, 129.03, 128.58, 127.82, 127.67, 127.44, 127.06, 125.61, 125.12, 123.58, 123.26, 119.95, 80.18, 80.08, 70.49, 70.42, 70.13, 70.06, 67.14, 66.58, 52.11, 51.92, 51.73, 50.90, 50.81, 47.25, 41.59, 40.90, 39.06, 36.85, 28.52 ppm. 15のLC-MS:LC-MS (m / z): C 51 H 62 N7O 10 [M+H] calculated value: 932.45, measured value 932.45g / mol. HRMS-ESI: C 51 H 61 N7O 10 Calculated value of Na [M + Na]+: 954.4377, measured value 954.4433.

[0751] [Example 29] Synthesis of (9H-fluoren-9-yl)methyl(1-(4-(1-((2-(2-((3S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidine-3-yl)acetyl)isoindorin-4-yl)methyl)-1H-1,2,3-triazole-4-yl)phenyl)-1,6-dioxo-9,12,15,18-tetraoxa-2,5-diazaeicosan-20-yl)carbamate (compound 16) To a stirred solution of intermediate 15 (250 mg, 0.268 mmol) in DCM (1.0 mL), TFA (0.5 mL) was added at room temperature. This mixture was continuously stirred for 30 minutes, after which the solvent was evaporated and the mixture was dried under vacuum. In separate round-bottom flasks, acid intermediate 5 (100 mg, 0.333 mmol), followed by HATU (151 mg, 0.399 mmol) and DIPEA (0.170 mL, 0.999 mmol) were dissolved in DMF (0.5 mL). To activate the acid functional groups, the reaction mixture was stirred under a nitrogen atmosphere at room temperature for 10 minutes.

[0752] The amine generated in situ from intermediate 15 (220 mg, 0.268 mmol) was dissolved in DMF (1 mL) and added to the above reaction mixture, and continuously stirred for 2 hours. Next, this reaction mixture was diluted with water (15 mL) and stirred at room temperature for 15 minutes. The black suspension (turbidity) formed in the reaction mixture was filtered and redissolved in a mixture of methanol and dichloromethane. The organic layer was evaporated, and the resulting crude residue was purified by reverse-phase preparative high-performance liquid chromatography (HPLC) (A=20 mm ammonium acetate buffer (pH=7), B=acetonitrile, B5% to 95% solvent gradient) for 60 minutes to obtain compound 16 as a white solid (175 mg, 65%). Figures 34A-34B show data supporting the formation of compound 16, and Figure 34C shows H supporting the formation of FAP-4003. 1 The NMR (D2O) data is shown. 1H-NMR (CD3OD + CDCl3, 500 MHZ) δ = 8.32 (d, J= 15.0 Hz, 1H), 7.89-7.85 (m, 4H), 7.73 (d, J = 10.0 Hz, 2H); 7.58 (d, J = 5.0 Hz, 2H); 7.35-7.24 (m, 8H); 5.61-5.59 (m, 2H); 5.05 (dd, J1 = 9.1 Hz, J2 = 3.9 Hz, 1H); 4.85 (m, 2H); 4.72 (d, J = 17.2Hz, 2H); 4.45-4.37 (m, 1H); 4.31 (d, J = 6.8 Hz, 2H); 4.15-4.02 (m, 3H); 3.67 (t, J = 6.0 Hz, 2H); 3.55-3.48 (m, 20H); 3.31-3.25 (m, 6H); 3.0-2.74 (m, 5H); 2.60-2.54 (m, 1H); 2.43-2.30 (m, 4H) ppm. 13 C-NMR (CD3OD + CDCl3, 125 MHZ) δ = 173.28, 168.36, 156.66, 143.88, 141.19, 127.76, 127.45, 126.83, 125.34, 124.82, 121.72, 119.62, 70.72, 70.19, 69.99, 69.91, 69.62, 66.85, 66.30, 52.80, 52.06, 44.58, 40.44, 39.86, 38.69, 37.60 36.38 ppm. 16のLC-MS: LC-MS (m / z): C 58 H 65 F2N 10 O 11 [M+H] calculated value: 1115.47, measured value 1115.47 g / mol. HRMS-ESI: C 58 H 65 F2N 10 O 11 Calculated value of [M+H]: 1115.4802, measured value 1115.4817.

[0753] [Example 30] Sodium 2-((E)-2-((E)-2-(4-(1-(4-(1-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4-difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidine-3-yl)acetyl)isoindoline-4-yl)methyl)-1H-1,2,3-triazole-4-yl)phenyl)-1,6,22-trioxo-9,12,15,18-tetra Synthesis of Oxa-2,5,21-Triazatetracosan-24-yl)phenoxy)-3-(2-((E)-3,3-dimethyl-5-sulfonate-1-(4-sulfonatebutyl)indoline-2-ylidene)ethylidene)cyclohexa-1-en-1-yl)vinyl)-3,3-dimethyl-1-(4-sulfonatebutyl)-3H-indole-1-ium-5-sulfonate (FAP-4003) To a stirred solution of compound 16 (250.0 mg, 0.2244 mmol) in DCM+MeOH (1+1 mL), diethylamine (Et)2NH (2.0 mL) was added and the mixture was continuously stirred at room temperature for 2 hours. The reaction mixture was evaporated under reduced pressure, and the resulting crude residue was redissolved in SiO (10 mL). The resulting precipitate was filtered through a Buchner funnel, yielding free amine (intermediate 2) as a brown solid. LC-MS of 2: LC-MS (m / z): C 43 H 55 F2N 10 Calculated value of O9 [M+H]: 893.40, measured value: 893.40 g / mol. HRMS-ESI: C 43 H 55 F2N 10 Calculated value of O9 [M+H] + : 893.4121, measured value 893.4160. This substance was used in the synthesis of FAP-targeted dye compound 4, described below.

[0754] Next, to a stirred solution of amine (intermediate 2) (50 mg, 0.056 mmol) in DMF (1.0 mL), 3-(4-hydroxyphenyl)propionic acid (14.0 mg, 0.084 mmol), HATU (38.0 mg, 0.101 mmol), and DIPEA (42 μl, 0.252 mmol) were added to obtain 4-(1-((2-(2-((3S,5S)-5-((S)-2-cyano-4,4 Intermediate 3 was prepared by preparing -difluoropyrrolidine-1-carbonyl)-2-oxopyrrolidine-3-yl)acetyl)isoindoline-4-yl)methyl)-1H-1,2,3-triazole-4-yl)-N-(22-(4-hydroxyphenyl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazadocosyl)benzamide (intermediate 3), and the mixture was continuously stirred at room temperature for 30 minutes. Next, the reaction mixture was quenched with water (5 mL), and the resulting crude residue was purified by UHPLC (A = 20 Mm ammonium acetate buffer (pH = 7), B = acetonitrile, solvent gradient from 5% to 95% over 60 minutes) to obtain intermediate 3 as a white solid (34 mg, 60%). LC / MS of 3: LC-MS (m / z): C52 H 63 F2N 10 O 11 Calculated value [M+H]+: 1041.46, measured value: 1041.46. HRMS-ESI: C 52 H 63 F2N 10 O 11 The calculated value [M+H]+ is 1041.4645, and the measured value is 1041.4650.

[0755] To a stirred solution of intermediate 3 (5.0 mg, 0.00480 mmol) in anhydrous DMSO (500 μl), ClS0456 dye (4.6 mg, 0.0048 mmol) and then Cs2CO3 (15.0 mg, 0.0480 mmol) were added under an argon atmosphere at room temperature. This mixture was continuously stirred at room temperature for a further 3-4 hours, and the progress of the reaction was monitored by LC-MS.

[0756] Next, this reaction mixture was diluted with water and purified by UHPLC (A = 20 mm ammonium acetate buffer (pH = 7), B = acetonitrile 0, solvent gradient from 5% to 35% B over 60 minutes), yielding intermediate 5 as a cottony green solid (6.0 mg, 66%). LC-MS of 4: LC / MS (m / z): [M + H] C 90 H 110 F2N 12 O 23 Calculated value of S4[M+H]: 1892.66, measured value: 1892.60, [M+H] / 2: 946.0, and [M+H] / 3: 630.0 g / mol. 1H-NMR (D2O, 500 MHZ) δ = 8.27 (m, 1H); 7.62 (m, 10H); 7.12 (d, J = 6.2Hz, 5H); 6.97 (dd, J = 14.7 Hz, 7.9 Hz, 2H); 6.64 (dd, J = 14.1 Hz, 8.1 Hz, 2H); 5.86 (dd, J = 14.3 Hz, 3H); 5.40 (d, J = 9.8 Hz, 3H);4.97 (m, 3H); 4.54 (d, J = 7.5 Hz, 3H); 4.46 (s, 1H); 4.35 (s, 1H); 3.97 (m, 1H); 3.85 (s, 6H); 3.50 (s, 6H); 3.26 (m, 20H); 3.09 (m, 3H); 2.77(s, 10H); 2.62 (m, 4H); 2.33 (m,9H); 2.15 (m, 2H); 1.90 (d, J = 0.9 Hz, 7H); 1.68 (s, 10 H); 0.95 (s, 10 H) ppm.

[0757] Figure 35 shows the excitation and emission spectra of a 1 μM FAP-4003 solution in PBS at pH 7.4 after synthesis. Furthermore, this disclosure includes the following aspects. [Aspect 1] Structure of equation (X): A m -L-B' (X) (In the formula, A is given by equation XB: [ka] It is the radical of the fibroblast-activating protein alpha (FAPα) ligand, During the ceremony, T is a substituted or unsubstituted methylene group (-CH 2 -), substituted or unsubstituted amino(-NH-), -O- or -S- (for example, substitution of T is C 1 ~C 3 (It is alkyl, haloalkyl, or halo) J is C(R J ) 0~3 And R J Each of these is independently either H or alkyl, or two or more R J They come together to form an oxo, R 1 and R 2 is -H, -CN, -CHO, -B(OH) 2 -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O) 2 Aryl, -CO 2 H, -SO 3 H, -SO 2 NH 2 ,-PO 3 H 2 , -SO 2 F, -CONH 2 and independently selected from the group consisting of 5-tetrazolyl, R 3 and R 4 -H, -OH, F, Cl, Br, I, -C 1~6 Alkyl, -OC 1~6 Alkyl and -SC 1~6 Independently selected from the group consisting of alkyls, R 5 、R 6 、R 7 and R 8 It is independently selected from the group consisting of H, alkyl, and halo, R 9 、R 10 and R 11 H, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, -SC 1~6 Independently selected from the group consisting of alkyl, F, Cl, Br, and I, L is a linker that connects A to B', B' is a radical of a therapeutic agent, radioimaging agent, radiotherapy agent, magnetic resonance imaging agent, chemotherapy agent, antifibrotic agent, or anticancer agent. (m = 1 to 6) A compound represented by [this symbol]. [Aspect 2] It also contains C', L links C' to one or more A groups and B', C' is an albumin-binding ligand, polyethylene glycol n (PEG) n The compound according to embodiment 1, wherein n is an integer from 0 to 32, and is a peptide, peptidoglycan, or sugar radical. [Aspect 3] The compound according to embodiment 1 or 2, wherein B' is a phosphoinositide 3-kinase (PI3K) inhibitor, a chelating group optionally bound to an isotope (or metal), or a radical of a group covalently bonded to an isotope (or metal), and the isotope or metal is suitable for radiation imaging, radiation therapy, or magnetic resonance imaging. [Aspect 4] Compounds represented by the structure of formula (I'):

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Claims

1. Structure of formula (V): 【Transformation 36】 (In the formula, L is a linker containing at least one carbon atom, p is 0, 1, 2, or 3. L, 【Transformation 38】 And, m is an integer from 1 to 9, n is an integer between 1 and 32. q is an integer between 0 and 4, (where s is an integer between 0 and 4) A compound represented by [this symbol]. 【Request Item 2】 【Chemistry 39】 (In the formula, t is 0 or 1, and u is an integer between 2 and 12.) A compound that is 【Request Item 3】 【Chemistry 40】 (m is an integer between 1 and 4) A compound that is 【Request Item 4】 【Chemistry 41】 A compound that is 【Request Item 5】 【Chemistry 42】 A compound that is 【Request Item 6】 【Chemistry 43】 A compound that is 【Request Item 7】 【Chemistry 44】 A compound that is 【Request Item 8】 【Chemistry 45】 A compound that is

9. The following structure: 【Chemistry 51】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

10. The following structure: 【Transformation 65】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

11. The following structure: 【Chemical Formula 66】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

12. The following structure: 【Transformation 67】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

13. The following structure: 【Chemistry 80】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

14. The following structure: 【Chemistry 81】 A compound that is optionally bound to an isotope suitable for radiation imaging, radiotherapy, or magnetic resonance imaging.

15. structure: 【Chemistry 93】 A compound having the following properties.

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15 and a pharmaceutically acceptable carrier.

17. A compound according to any one of claims 1 to 15 for use in imaging cancer or fibrosis in a subject.

18. The pharmaceutical composition according to claim 16 for use in imaging cancer or fibrosis in a subject.

19. The compound according to claim 17, wherein the imaging comprises generating an image of the cancer or fibrosis in the subject.

20. The pharmaceutical composition according to claim 18, wherein the imaging comprises generating an image of the cancer or fibrosis in the subject.

21. The compound according to claim 17, wherein the fibrosis is selected from pulmonary fibrosis, renal fibrosis, and hepatic fibrosis.

22. The pharmaceutical composition according to claim 18, wherein the fibrosis is selected from pulmonary fibrosis, renal fibrosis, and hepatic fibrosis.

23. A compound according to any one of claims 1 to 15, for use in the treatment of fibrosis in a subject.

24. The pharmaceutical composition according to claim 16, for use in the treatment of fibrosis in a subject.

25. The compound according to claim 23, wherein the fibrosis is selected from pulmonary fibrosis, renal fibrosis, and hepatic fibrosis.

26. The pharmaceutical composition according to claim 24, wherein the fibrosis is selected from pulmonary fibrosis, renal fibrosis, and hepatic fibrosis.

27. A compound according to any one of claims 1 to 15 for use in treating inflammatory diseases or disorders in subjects requiring such treatment.

28. The pharmaceutical composition according to claim 16 for use in treating inflammatory diseases or disorders in subjects requiring the same.

29. A compound according to any one of claims 1 to 15, for use in the treatment of cancer in a subject requiring it.

30. The pharmaceutical composition according to claim 16 for use in the treatment of cancer in a subject requiring it.

31. The compound according to claim 29, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, colorectal cancer, cervical cancer, and brain cancer.

32. The pharmaceutical composition according to claim 30, wherein the cancer is selected from the group consisting of lung cancer, breast cancer, colorectal cancer, cervical cancer, and brain cancer.

33. The compound according to claim 29 or 31, for further use in combination with chemotherapy or radiotherapy applied to the subject.

34. The pharmaceutical composition according to claim 30 or 32, for further use in combination with chemotherapy or radiotherapy applied to the subject.

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