FAP-Activated Radiotheranostics and Related Uses

Abstract

The present disclosure relates to fibroblast activation protein (FAP)-activated theranostic prodrugs, pharmaceutical compositions containing them, and methods of treating disorders characterized by upregulation of FAP, such as cancer.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 126,617, filed December 17, 2020, which is incorporated by reference herein in its entirety. [Background technology]

[0002] Fibroblast activation protein alpha (FAP), also known as seprase, is a type II integral membrane serine peptidase. FAP belongs to the dipeptidyl peptidase IV family. It is a 170 kDa homodimer containing two N-glycosylated subunits with a large C-terminal extracellular domain in which the catalytic domain of the enzyme is located. In its glycosylated form, FAP possesses both post-prolyl dipeptidyl peptidase and gelatinase activities. Homologues of human FAP have been found in several species, including mice and cynomolgus monkeys.

[0003] FAP is selectively expressed in reactive stromal fibroblasts in over 90% of epithelial malignancies (primary and metastatic) examined, including lung, colorectal, bladder, ovarian, and breast carcinomas, and in malignant mesenchymal cells of osteosarcoma and soft tissue sarcomas, but is generally absent from normal adult tissues (Non-Patent Documents 1-4). FAP is also expressed in certain malignant tumor cells.

[0004] Due to its expression in many common cancers and its restricted expression in normal tissues, FAP is considered a promising antigenic target for imaging, diagnosis, and treatment of various cancers. Various approaches have been devised to exploit the selective expression of FAP in the tumor stroma for clinical benefit, including monoclonal antibodies against FAP, small molecule inhibitors of FAP enzymatic activity, FAP-activated prodrugs of cytotoxic compounds, and FAP-specific CAR-T cells. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Brennen et al., Mol. Cancer Ther. 11 (2): 257-266 (2012) [Non-patent document 2] Garin-Chesa et al., Proc Natl Acad Sci USA 87, 7235-7239 (1990) [Non-patent document 3] Rettig et al., Cancer Res. 53:3327-3335 (1993) [Non-patent document 4] Rettig et al., Proc Natl Acad Sci USA 85, 3110-3114(1988) Summary of the Invention [Means for solving the problem]

[0006] FAP-activated radiotheranostics are disclosed that enable the selective delivery of radioactive diagnostic and therapeutic agents to the tumor microenvironment. These include radiotherapeutic agents designed to target other molecules or receptors in the tumor microenvironment, such as prostate-specific membrane antigen, folate receptors, and somatostatin. FAP activation reduces exposure to normal cells and tissues that express or contain significant levels of the targeted key receptors or molecules, thereby reducing adverse side effects and improving the therapeutic window and efficacy.

[0007] One aspect of the present invention relates to FAP-activated theranostic prodrugs and compositions containing them. Another aspect of the present invention is a method of using the prodrugs and compositions containing them to treat disorders characterized by upregulation of fibroblast activation protein (FAP).

[0008] In certain embodiments, the subject FAP-activated theranostic prodrug agents have the general formula I: [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, "FAPs" refers to a moiety containing a FAPα substrate ("FAP substrate moiety") that is cleaved by FAPα to release FAPs-C(=O)OH and NH2-LR; L is a bond or a self-eliminating linker that upon cleavage by FAP releases NH2-LR; R represents a ligand-targeting theranostic moiety that includes a ligand for binding to a cellular target and one or more of a radioactive moiety and / or a chelator for chelating the radioactive moiety.

[0009] In some embodiments, enzymatic cleavage of the prodrug by a fibroblast activation protein (FAP) results in release of the ligand-targeted theranostic moiety either as an activated ligand-targeted theranostic moiety (i.e., its pharmacologically active form) or in a form that is readily metabolized to its active form; and, upon release from the prodrug by FAP cleavage, the activated ligand-targeted theranostic moiety binds to the cellular target with a Kd for binding to the cellular target that is less than the Kd for binding of the prodrug to the cellular target (i.e., has a higher affinity for the cellular target).

[0010] In some embodiments, the FAP-activated theranostic prodrug is Formula II: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, R represents a ligand-targeting theranostic moiety that includes a ligand for binding to a cellular target and one or more of a radioactive moiety and / or a chelator for chelating the radioactive moiety; A represents a 5- to 8-membered heterocycle; X is O or S; R 10 is an amino terminal blocking group; R 12 is hydrogen or (C1-C6) alkyl; R 13 is hydrogen, (C1-C6) alkyl (which may be linear or branched) or (C1-C6); R 14 is, independently for each occurrence, -(C1-C6)alkyl, -OH, -NH2, or halogen; p is an integer from 0 to 6; L is a bond or a self-eliminating linker that upon cleavage by FAP releases NH2-LR; Enzymatic cleavage of the prodrug by fibroblast activation protein (FAP) results in release of the ligand-targeted theranostic moiety either as an activated ligand-targeted theranostic moiety (i.e., its pharmacologically active form) or in a form that is readily metabolized to that active form; and, upon release from the prodrug by FAP cleavage, the activated ligand-targeted theranostic moiety binds to the cellular target with a Kd for binding to the cellular target that is less than the Kd for binding of the prodrug to the cellular target (i.e., has a higher affinity for the cellular target).

[0011] In certain preferred embodiments of Formula II, R 12 is H.

[0012] In certain embodiments of the structure of formula II, R 10 forms an amide or thioamide with the nitrogen to which it is attached, and the prodrug has the formula IIa: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, A, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0013] In certain preferred embodiments of Formula IIa, X is O; and / or R 12 is H. In certain preferred embodiments, X is O and R 12 is H.

[0014] In certain embodiments, a preferred FAP-activated theranostic prodrug has Formula III: [ka] or a pharmaceutically acceptable salt thereof, wherein R, R 10 , R 12 , R 13 , R 14 , L, X and p are as defined above for Formula II.

[0015] In certain embodiments of the structure of formula III, R 10 forms an amide or thioamide with the nitrogen to which it is attached, and the prodrug has the formula IIIa: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0016] In certain embodiments, the FAP-activated theranostic prodrug has Formula IV: [ka] or a pharmaceutically acceptable salt thereof, wherein R 13is (C1-C6) alkyl (which may be linear or branched) or (C1-C6), and R, R 10 , R 12 , R 13 , R 14 , L, X and p are as defined above for Formula II.

[0017] In certain embodiments of the structure of formula IV, R 10 forms an amide or thioamide together with the nitrogen to which it is attached, and the prodrug has the formula IVa: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n-(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0018] In certain embodiments, R 13 is C1-C6 alkyl, such as methyl. In another embodiment, R 13 is hydrogen.

[0019] In certain embodiments, R 12 is H.

[0020] In certain embodiments, p is 1 or 2 and R 14 is a halo for each occurrence. In other embodiments, p is 0.

[0021] In some embodiments, the FAP-activated radiopharmaceutical has Formula V: [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; In the formula, R, R 10 , R 11 , R 13 and L is as described for formula II.

[0022] In certain embodiments, the FAP-activated theranostic prodrug has Formula VI: [ka] or a pharmaceutically acceptable salt thereof; [ka] or a pharmaceutically acceptable salt thereof; In the formula, R, R 10 , R 11 and L is as described for formula II.

[0023] In yet other embodiments, the FAP-activated theranostic prodrug has Formula VII: [ka] or a pharmaceutically acceptable salt thereof, wherein R and L are as defined above for formula II; and 11 forms an acyl group.

[0024] In some embodiments, X is O.

[0025] In some embodiments, R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 )alkoxy, -(C3-C8)cycloalkyl, -(C6-C 14 ) Aryl, Aryl (C1-C 10 ) alkyl, or 5- to 10-membered heteroaryl.

[0026] In some embodiments, R 11 teeth, [ka] is.

[0027] In some embodiments, n is equal to 1 and AA is a serine residue.

[0028] In some embodiments, R 11 is (C1~C 10 ) alkyl, (C1-C 10 ) alkoxy, (C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, (C3-C8) cycloalkyl, (C3-C8) cycloalkyl(C1-C 10 ) Alkyl, (C6-C 14 ) Aryl, Aryl (C1-C 10 ) alkyl, 5-10 membered heteroaryl, or 5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; R 12 is hydrogen; R 13 is (C1-C6) alkyl; R 14 does not exist, or p is 2 and R 14 is a halogen for each occurrence; L is a bond or -N(H)-L- is a self-elimination linker.

[0029] In some embodiments, —C(O)—R 11 forms an acyl group of the carboxylic acid, such as a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0030] In other embodiments, R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, preferably a 6 membered aryl or heteroaryl group, and m is an integer from 1 to 6, preferably 1 or 2. In some embodiments, aryl is selected from the group consisting of benzyl, naphthalenyl, phenanthrenyl, phenolyl, and anilinyl. In other embodiments, heteroaryl is selected from the group consisting of pyrryl, furyl, thiophenyl (a / k / a thienyl), imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl.

[0031] In some embodiments, L is a bond, while in other embodiments, L is a self-elimination linker. [ka] [ka] may be selected from the group consisting of During the ceremony, R a is hydrogen, alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl; R b is halogen, alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl; h is an integer from 0 to 8, as valence permits; i is an integer from 1 to 6.

[0032] In other embodiments, L is [ka] where R 1 is hydrogen, unsubstituted or substituted C 1-3 It is alkyl, or unsubstituted or substituted heterocyclyl.

[0033] In still other embodiments, L is [ka] is selected from.

[0034] In some embodiments, L is [ka] is selected from During the ceremony, U is O, S or NR 6 and; Q is CR 4 or N; V 1 , V 2 and V 3 independently, CR 4 or N, with the proviso that for Formulas II and III, Q, V 1 and V 2 at least one of is N; T is NH, NR pending from the therapeutic moiety 6 , O or S: R 1 , R 2 , R 3 and R 4are independently H, F, Cl, Br, I, OH, -N(R 5 )2, -N(R 5 )3 + , C1 to C8 halogenated Alkyl (or alkylhalide), carboxylate, sulfate, sulfamate, sulfonate, -SO2R 5 , -S(=O)R 5 , -SR 5 , -SO2N(R 5 )2, -C(=O)R 5 , -CO2R 5 , -C(=O)N(R 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 halosubstituted alkyl, polyethyleneoxy, phosphonate, phosphate, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocycles and C1-C 20 substituted heterocycles; or R 2 and R 3 together form a carbonyl (=O) or a spirocarbocyclic ring of 3 to 7 carbon atoms; R 5 and R 6 are independently H, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocycles and C1-C 20 substituted heterocycles; where C1 to C8 substituted alkyl, C2 to C8 substituted alkenyl, C2 to C8 substituted alkynyl, C6 to C 20 Substituted aryl and C2-C 20 Substituted heterocycles are independently selected from F, Cl, Br, I, OH, N(R 5 )2, -N(R 5 )3+ , C1-C8 alkyl halide, carboxylate, sulfate, sulfamate, sulfonate, C1-C8 alkyl sulfonate, C1-C8 alkyl amino, 4-dialkylaminopyridinium, C1-C8 alkyl hydroxyl, C1-C8 alkyl thiol, -SO2R 5 , -S(=O)R 5 , -SR 5 , -SO2N(R 5 )2, -C(=O)R 5 , -CO2R 5 , -C(=O)N(R 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 trifluoroalkyl, C1-C8 alkyl, C3-C 12 Carbocycle, C6~C 20 Aryl, C2-C 20 It is substituted with one or more substituents selected from heterocycle, polyethyleneoxy, phosphonate, and phosphate.

[0035] In some embodiments, L is selected from -NH-(CH2)4-C(=O)-, or -NH-(CH2)3-C(=O)-, p-aminobenzyloxycarbonyl (PABC), 2,4-bis(hydroxymethyl)aniline, or benzyloxycarbonyl.

[0036] In some embodiments, the ligand-targeting theranostic moiety (R) is -TM-L 1 -R 20 Represented by: During the ceremony, TM represents a ligand targeting moiety that selectively binds to a cell surface feature on a target cell; L 1 represents a bond or a linker; R 20represents a radioactive moiety, a chelator, a fluorescent moiety, a photoacoustic reporting molecule, a Raman-active reporting molecule, an imaging agent, or a detectable nanoparticle.

[0037] In some embodiments, the ligand targeting moiety is a folate receptor ligand, preferably folic acid or a folic acid analog, preferably etarfolatide, vintafolide, leucovorin, and methotrexate.

[0038] In some embodiments, the ligand targeting moiety is somatostatin or a somatostatin analog, preferably octreotate, octreotide, pentetreotide, lanreotide, vapreotide, pasireotide, seglitide, benereotide, KE-108, SDZ-222-100, Sst3-ODN-8, CYN-154806, JR11, J2156, SRA-880, ACQ090, P829, SSTp-58, SSTp-86, BASS, or somatoprim.

[0039] In yet other embodiments, the ligand targeting moiety is an αIIbβ3 targeting ligand such as RGD or an RGD analog, preferably cyclo(-Arg-Gly-Asp-D-PheVal-)["c(RGDfV)"], c(RGDfK), c(RGDfC), c(RADfC), c(RADfK), c(RGDfE), c(RADfE), RGDSK, RADSK, RGDS, c(RGDyC), c(RADyC), c(RGDyE), c(RGDyK), c(RADyK), HE[c(RGDyK)]2, EMD12194, DMP728, DMP757, and SK&F107260.

[0040] In some embodiments, the targeted theranostic moiety (R) is [ka] where R 30 represents independently for each occurrence hydrogen or lower alkyl.

[0041] In some embodiments, -L 1 -R 20 teeth, [ka] wherein R 31 Ha-(CH2) p -aryl or -(CH2) p -heteroaryl, and p is 0, 1, 2, 3, or 4.

[0042] In some embodiments, R 31 is -CH2-aryl, where the aryl group is C6-C 12 Aryl is a monocyclic or bicyclic fused ring, preferably naphthalene, e.g., -L 1 -R 20 teeth, [ka] It can be expressed as:

[0043] In some embodiments, R 20 is F 18 containing moiety, e.g., -L 1 -R 20 teeth, [ka] may be selected from:

[0044] In some embodiments, the ligand-targeting theranostic moiety (R) comprises a folic acid or folic acid analog selected from the following: [ka] During the ceremony, R 21 represents H, and R 22 is -NH-(CH2) q -R 20 , -NH-(CH2) q -NH-C(O)-(CH2) q -R 20 , or -NH-(CH2) q -C(O)-(CH2) q -R 20 represents; or R 22 represents H, and R 21 is -NH-(CH2) q -R 20 or -NH-(CH2) q -C(O)-(CH2) q -R 20 represents; or R 21 or R 22 One of them represents H and the other represents [ka] selected from the group consisting of: R 23 represents H, -CH3, -CH2CH3, or -CO2H; q is independently 0, 1, 2, 3, or 4 for each occurrence.

[0045] In other embodiments, R 21 is -NH-CH2-R 20 , -NH-CH2-C(O)- R 20 , -NH-C(O)-CH2-R 20 , -NH-CH2-C(O)-CH2-R 20 , or -NH-(CH2)2-NH-C(O)-CH2-R 20 represents R 22 represents H.

[0046] In yet other embodiments, R 21 represents H, and R 22is -NH-CH2-R 20 , -NH-CH2-C(O)- R 20 , -NH-C(O)-CH2-R 20 , -NH-CH2-C(O)-CH2-R 20 , or -NH-(CH2)2-NH-C(O)-CH2-R 20 Represents.

[0047] In some embodiments, the ligand-targeting theranostic moiety (R) is [ka] is.

[0048] In yet other embodiments, the ligand-targeting theranostic moiety (R) comprises a radiolabeled folic acid or folic acid analog selected from: [ka] In the formula, R 23 represents H, -CH3, -CH2CH3, or -CO2H, and X is CR 40 or N, R 40 is H or lower alkyl.

[0049] In some embodiments, R is [ka] [ka] is selected from.

[0050] In some embodiments, R 20 teeth, [ka] is selected from.

[0051] In yet other embodiments, R 20 -L 1 -teeth, [ka] is.

[0052] In some embodiments, R is a ligand for an extracellular receptor. For example, R is a ligand for an extracellular receptor that undergoes intracellular internalization, such that R, upon release from the prodrug, can be transported to one or more intracellular compartments of cells expressing the extracellular receptor. In yet other embodiments, the cellular target is expressed by cells in a tissue in which FAP expression is upregulated. In a further embodiment, the tissue in which FAP expression is upregulated is a tumor.

[0053] In some embodiments, R is an analog, such as a peptide analog, that binds to a peptide hormone receptor. For example, R can be an analog, such as a peptide analog of somatostatin, bombesin, calcitonin, oxytocin, EGF, α-melanocyte-stimulating hormone, minigastrin, neurotesin, or neuropeptide (or neuropeptide) Y (NPY).

[0054] In some embodiments, R is a ligand that binds to integrin αβ, gastrin-releasing peptide receptor (GRPR), somatostatin receptor (such as somatostatin receptor subtype 2), melanocortin receptor, cholecystokinin-2 receptor, neuropeptide Y receptor, or neurotensin receptor.

[0055] In some embodiments, R is a ligand that binds to a type II membrane protein, such as prostate-specific membrane antigen (PSMA). For example, theranostic prodrugs include: [ka] [ka] [ka] or a pharmaceutically acceptable salt thereof, which may optionally include a radioisotope chelated thereto.

[0056] In some embodiments, R is a ligand that binds to a somatostatin receptor, e.g., [ka] or a pharmaceutically acceptable salt thereof, which may optionally contain a radioisotope chelated thereto.

[0057] In some embodiments, the ligand is 18 F, 43 K. 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 75 Br, 76 Br, 77 As, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99m Tc, 100 Pd, 101m Rh, 103 Pb, 105 Rh, 109 Pd, 111 Ag, 111 In, 113 In, 119 Sb 121 Sn, 123 I, 124 I, 125 I, 127 Cs,128 Ba, 129 Cs, 131 Cs, 131 I, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 EU, 153 EU, 153 Sm, 159 Gr, 161 Tb, 165 Dy, 166 Ho, 169 EU, 175 Yb, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, and 227 The present invention includes chelating agents that chelate or are capable of chelating radioactive metal or semi-metal isotopes such as Th.

[0058] In some embodiments, the prodrug has a k for cleavage by a FAP that is at least 10 times greater than for cleavage by a prolyl endopeptidase. cat / K m It has.

[0059] The present disclosure also provides a pharmaceutical composition comprising a FAP-activated theranostic prodrug of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0060] The present disclosure also provides a method of treating a disorder characterized by upregulation of fibroblast activation protein (FAP), comprising administering to a subject in need thereof a therapeutically effective amount of a FAP-activating theranostic prodrug of any one of the preceding claims, or a pharmaceutically acceptable salt thereof. In some embodiments, the disorder characterized by upregulation of FAP is cancer.

[0061] In a further aspect, there is provided a method of treating a subject having prostate cancer, which may suitably comprise administering to a subject in need thereof a therapeutically effective amount of a FAP-activated theranostic prodrug of any one of the preceding claims, or a pharmaceutically acceptable salt thereof.

[0062] Other aspects of the invention are disclosed below. [Brief explanation of the drawings]

[0063] [Figure 1] 1 shows a synthesis scheme for compound 7885. [Figure 2] 1 shows a synthesis scheme for compound 6885. [Figure 3] 1 shows a synthesis scheme for compound 6879. [Figure 4] 1 shows a synthesis scheme for compound 6880. [Figure 5] 1 shows a synthesis scheme for compound 6886. [Figure 6] 1 is a graph showing the results of an assay of PSMA activity for compounds 7028P-7028A / B / C. Data are the average of triplicate readings (standard error of the mean is plotted but not visible). [Figure 7] 1 shows a synthesis scheme for compound 6970B ester. [Figure 8] The synthesis scheme of compound 7014 is shown. [Figure 9] 1 shows a synthesis scheme for compound 7366P5. [Figure 10] 1 shows a synthesis scheme for compound 7366. [Figure 11] 1 is a graph showing the results of FAP activation of 6970B isomers 1 & 2 and 7366 crude product (Crude) using 100 μM substrate and 50 nM FAP. [Figure 12] LC / MS spectrum of 6970B isomer 1 is shown. [Figure 13] LC / MS spectrum of 6970B isomer 2 is shown. [Figure 14] LC / MS spectra of 6970B isomers 1 & 2 are shown. [Figure 15] The LC / MS spectrum of 7366 is shown. DETAILED DESCRIPTION OF THE INVENTION

[0064] I. overview Targeting the tumor microenvironment with FAP-activated radiopharmaceutical prodrugs is thought to have multiple antitumor effects, primarily relying on the induction of DNA damage in tumor cells by ionizing radiation locally emitted from adjacent CAFs targeted by the therapy. FAP-activated radiotherapy can deliver ionizing radiation to cancer cells and the tumor stroma. Combining α- and β-emitters can improve these dual antitumor effects through short-range α-irradiation of CAFs and medium- to long-range β-irradiation of cancer cells.

[0065] FAP-positive CAFs are found in over 90% of epithelial cancers and therefore represent a potential pan-cancer prodrug-activating enzyme. Targeting ligand-directed radiopharmaceuticals (and other theranostic agents) to tumors by creating FAP-activated prodrug versions is a means of delivering activated ligand-directed radiopharmaceuticals selectively to tumors in a form that can bind to the cellular targets to which the natural ligand binds after being cleaved by FAP in the tumor. Because the circulating prodrug form has significantly reduced binding to its cellular targets compared to the ligand released from the prodrug by FAP cleavage, it is taken up to a lesser extent by non-tumor tissues (salivary glands, kidneys, etc.) than the activated ligand-directed radiopharmaceutical, which can result in an enhanced therapeutic index, better efficacy, or both of the prodrug (compared to when the activated ligand-directed radiopharmaceutical is administered in its original form).

[0066] In certain embodiments, the subject FAP-activated theranostic prodrug agents have the general formula: [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, FAPs represents a moiety containing a FAPα substrate ("FAP substrate moiety") that is cleaved by FAPα to release FAPs-C(=O)OH and NH2-LR; L is a bond or a self-eliminating linker that upon cleavage by FAP releases NH2-LR; R represents a ligand-targeting theranostic moiety that includes a ligand for binding to a cellular target and one or more of a radioactive moiety and / or a chelator for chelating the radioactive moiety; Enzymatic cleavage of the prodrug by fibroblast activation protein (FAP) results in release of the ligand-targeting theranostic moiety either as an activated ligand-targeting theranostic moiety (i.e., its pharmacologically active form) or in a form that is readily metabolized to its active form; upon release from the prodrug by FAP cleavage, the activated ligand-targeting theranostic moiety binds to the cellular target with a Kd for binding to the cellular target that is less than the Kd for binding of the prodrug to the cellular target (i.e., has a higher affinity for the cellular target).

[0067] In certain embodiments, the FAP substrate portion has a kcat / Km for cleavage by FAPα that is at least 10-fold, at least 100-fold, 1000-fold, 5000-fold, or 10,000-fold greater than the kcat / Km for cleavage by prolyl endopeptidase (EC 3.4.21.26; PREP).

[0068] In certain embodiments, the activated ligand-targeting theranostic moiety (i.e., when released from the prodrug) has a Kd for binding to the cellular target that is at least 2-fold less than the Kd of the prodrug that binds to the cellular target, and more preferably at least 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, 500-fold, or 1000-fold less.

[0069] In certain embodiments, the prodrug may be further characterized by one or more of the following features: · the prodrug has a therapeutic index that is at least 2-fold greater, more preferably at least 5-fold, 10-fold, 50-fold, 100-fold, 250-fold, 500-fold, 1000-fold, 5000-fold, or even 10,000-fold greater than the therapeutic index of the activated ligand-targeting theranostic moiety itself (i.e., when administered in its active form); the activated ligand-targeting theranostic moiety is present at a higher local concentration, e.g., at least 2-fold higher, more preferably at least 5-fold, 10-fold, 100-fold, or even 1000-fold higher, in the target tissue, i.e., the tumor or other target tissue expressing the FAP, compared to the concentration of the circulating activated ligand-targeting theranostic moiety; · the maximum tolerated dose of the prodrug is at least 2-fold greater, and even more preferably at least 5-fold, 10-fold, 100-fold, or even 1000-fold greater, than the maximum tolerated dose of the activated ligand-targeted theranostic moiety when administered alone in its active form; the receptor-mediated uptake of the prodrug is at least 50% less than the receptor-mediated uptake of the activated ligand-targeting theranostic moiety, and even more preferably at least 60%, 70%, 80%, 90%, 95%, 98%, 99%, or even 99.9% less; and / or the cell permeability of the prodrug is at least 50% less than the cell permeability of the activated ligand-targeting theranostic moiety, and even more preferably at least 60%, 70%, 80%, 90%, 95%, 98%, 99%, or even 99.9% less; and / or The prodrug has a molecular weight of less than 5000 amu; The circulating half-life of the prodrug is at least 25% longer than the circulating half-life of the activated ligand-targeting theranostic moiety alone, and even more preferably at least 50%, 75%, 100%, 150%, 200%, 500%, 750%, or even 1000% longer.

[0070] II. definition The term "fibroblast activation protein (FAP)" as used herein is also known as "seprase." Both terms can be used interchangeably herein. Fibroblast activation protein is a homodimeric integral membrane protein with a dipeptidyl peptidase IV (DPPIV)-like fold, characterized by an α / β-hydrolase domain and an eight-bladed β-propeller domain.

[0071] As used herein, the term "SPECT" is an abbreviation for Single Photon Emission Computed Tomography.

[0072] As used herein, the term "PET" is an abbreviation for positron emission tomography.

[0073] As used herein, the term "CT" is an abbreviation for computed tomography.

[0074] As used herein, the term MRI is an abbreviation for magnetic resonance imaging.

[0075] As used herein, the term "SIRT" is an abbreviation for selective internal radiation therapy.

[0076] As used herein, the term "EDTA" is an abbreviation for ethylenediaminetetraacetic acid.

[0077] As used herein, the term "DOTA" is an abbreviation for 1,4,7,10-tetraazacyclododecane-1,4,7,10-N,N',N",N'''-tetraacetic acid.

[0078] As used herein, the term "DTPA" is an abbreviation for diethylenetriaminepentaacetic acid.

[0079] As used herein, the term metal "chelating agent" or "chelator" refers to a multidentate ligand that forms two or more separate coordinate bonds with a single central atom (especially a radioisotope).

[0080] The term "therapeutically effective amount" as used herein includes within its meaning a non-toxic but sufficient amount of the compound or composition used in the present invention to provide the desired therapeutic effect.The exact amount required varies from subject to subject, depending on factors such as the species being treated, the age, weight and general condition of the subject, coexisting diseases, the severity of the condition being treated, the specific drug being administered and the method of administration.Therefore, for any given case, the appropriate "effective amount" can be determined by those skilled in the art using only routine methods.

[0081] The term "radioactive moiety" as used herein refers to a molecular assembly carrying a radioactive nuclide. The nuclide is attached by either a covalent or coordinate bond that remains stable under physiological conditions. Examples of radioactive moieties include [I]-3-iodobenzoic acid or GaDOTA.

[0082] As used herein, a "fluorescent isotope" emits electromagnetic radiation after excitation by electromagnetic radiation of a shorter wavelength.

[0083] As used herein, a "radioisotope" is a radioactive isotope of an element (included in the term "radionuclide") that emits alpha, beta, or gamma radiation.

[0084] The term radiopharmaceutical is used in the context of the present invention to refer to a biologically active compound modified with a radioisotope. In particular, intercalating agents can be used to deliver radioactivity directly into close proximity to DNA (e.g., Hoechst-33258). 131 I-supported derivatives).

[0085] The terms "chelating agent" and "chelate" are used interchangeably in the context of the present invention and refer to molecules, often organic molecules, and often Lewis bases, that have two or more unshared electron pairs available for donation to a metal ion. The metal ion is usually coordinated to the chelating agent by two or more electron pairs. The terms "bidentate chelating agent," "tridentate chelating agent," and "tetradentate chelating agent" refer to chelating agents that have two, three, and four electron pairs readily available to simultaneously donate to the metal ion coordinated by the chelating agent, respectively. Typically, the electron pairs of a chelating agent form coordinate bonds with a single metal ion, but in certain instances, a chelating agent may form coordinate bonds with multiple metal ions, allowing for a variety of binding modes.

[0086] The term "fluorochrome" is used in the context of the present invention to refer to a compound that emits visible or infrared light after excitation with electromagnetic radiation of a suitable shorter wavelength. It will be understood by those skilled in the art that each fluorescent dye has a given excitation wavelength.

[0087] The term "contrast agent" is used in the context of the present invention to refer to a compound that increases the contrast of structures or fluids in medical imaging. Enhancement is achieved by absorbing electromagnetic radiation or by modifying the electromagnetic field.

[0088] As used herein, the term "paramagnetism" refers to paramagnetism induced by unpaired electrons in a medium. Paramagnetic materials induce a magnetic field when an external magnetic field is applied. Unlike diamagnetism, the induced magnetic field has the same direction as the external magnetic field, and unlike ferromagnetism, the magnetic field is not maintained in the absence of an external magnetic field.

[0089] The term "therapeutically effective amount" as used herein includes within its meaning a non-toxic but sufficient amount of the compound or composition used in the present invention to provide the desired therapeutic effect.The exact amount required varies from subject to subject, depending on factors such as the species being treated, the age, weight and general condition of the subject, coexisting diseases, the severity of the condition being treated, the specific drug being administered and the method of administration.Therefore, for any given case, the appropriate "effective amount" can be determined by those skilled in the art using only routine methods.

[0090] The term "alkyl" refers to a saturated straight or branched carbon chain. Preferably, the chain contains 1 to 10 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, such as methyl, ethyl, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, pentyl, or octyl. Alkyl groups may be substituted.

[0091] The term "heteroalkyl" refers to a saturated straight or branched carbon chain. Preferably, the chain contains 1 to 9 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, octyl, etc., interrupted one or more times, e.g., 1, 2, 3, 4, 5 times, by the same or different heteroatoms. Preferably, heteroatoms are selected from O, S, and N, e.g., -O-CH3, -S-CH3, -CH2-O-CH3, -CH2-O-CH2-CH3, -CH2-S-CH3, -CH2-S-CH2-CH3, -CH2-CH2-O-CH3, -CH2-CH2-O-CH2-CH3, -CH2-CH2-S-CH3, -CH2-CH2-S-CH3, -CH2-CH2-CH3, etc. Heteroalkyl groups may be substituted.

[0092] The terms "cycloalkyl" and "heterocycloalkyl," alone or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl," respectively, preferably having 3, 4, 5, 6, 7, 8, 9, or 10 atoms forming the ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. The terms "cycloalkyl" and "heterocycloalkyl" are also meant to include bicyclic, tricyclic, and polycyclic versions thereof. The term "heterocycloalkyl" preferably refers to a 5-membered saturated ring (at least one member is an N, O, or S atom and may contain one additional O or one additional N atom); a 6-membered saturated ring (at least one member is an N, O, or S atom and may contain one additional O, one additional N, or two additional N atoms); or a 9- or 10-membered saturated bicyclic ring (at least one member is an N, O, or S atom and may contain one additional N atom, two additional N, or three additional N atoms). "Cycloalkyl" and "heterocycloalkyl" groups may be optionally substituted. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, spiro[3,3]heptyl, spiro[3,4]octyl, spiro[4,3]octyl, spiro[3,5]nonyl, spiro[5,3]nonyl, spiro[3,6]decyl, spiro[6,3]decyl, spiro[4,5]decyl, spiro[5,4]decyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, and the like.Examples of heterocycloalkyl include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, 1,8 diazo-spiro-[4,5]decyl, 1,7 diazo-spiro-[4,5]decyl, 1,6 diazo-spiro-[4,5]decyl, 2,8 diazo-spiro[4,5]decyl, 2,7 diazo-spiro[4,5]decyl, 2,6 diazo-spiro[4,5]decyl, 1,8 diazo-spiro-[5,4]decyl, 1,7 diazo-spirotetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like.

[0093] The term "aryl" preferably refers to an aromatic monocyclic ring containing 6 carbon atoms, an aromatic bicyclic ring system containing 10 carbon atoms, or an aromatic tricyclic ring system containing 14 carbon atoms. Examples include phenyl, naphthyl, or anthracenyl. Aryl groups may be substituted.

[0094] The term "aralkyl" refers to an alkyl moiety substituted by an aryl, where alkyl and aryl have the meanings outlined above. An example is the benzyl radical. Preferably, in this context, the alkyl chain contains 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, such as methyl, ethylmethyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butenyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. The aralkyl group may be substituted in the alkyl and / or aryl portions of the group.

[0095] The term "heteroaryl" preferably refers to a 5- or 6-membered aromatic monocyclic ring in which at least one of the carbon atoms is replaced by 1, 2, 3, or 4 (in the case of a 5-membered ring) or 1, 2, 3, 4, or 5 (in the case of a 6-membered ring) heteroatoms, which may be the same or different, preferably selected from O, N, and S; an aromatic bicyclic ring system in which 1, 2, 3, 4, 5, or 6 of the 8, 9, 10, 11, or 12 carbon atoms are replaced by the same or different heteroatoms, preferably selected from O, N, and S; or an aromatic tricyclic ring system in which 1, 2, 3, 4, 5, or 6 of the 13, 14, 15, or 16 carbon atoms are replaced by the same or different heteroatoms, preferably selected from O, N, and S. Examples include oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,5-thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1-benzofuranyl, 2-benzofuranyl, indoyl, isoindoi. isoindoyl, benzothiophenyl, 2-benzothiophenyl, 1H-indazolyl, benzimidazolyl, benzoxazolyl, indoxazinyl, 2,1-benzoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl, 2,1-benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, quinolinyl, 1,2,3-benzotriazinyl or 1,2,4-benzotriazinyl.

[0096] The term "heteroaralkyl" refers to an alkyl moiety substituted by a heteroaryl, where alkyl and heteroaryl have the meanings outlined above. Examples include 2-alkylpyridinyl, 3-alkylpyridinyl, or 2-methylpyridinyl. Preferably, in this context, the alkyl chain contains 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, such as methyl, ethylmethyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butenyl, tert-butyl, pentyl, hexyl, pentyl, and octyl.

[0097] Heteroaralkyl groups may be optionally substituted on the alkyl and / or heteroaryl portions of the group.

[0098] The terms "alkenyl" and "cycloalkenyl" refer to an olefinically unsaturated carbon atom containing chain or ring with one or more double bonds. Examples include propenyl and cyclohexenyl. Preferably, the alkenyl chain contains 2 to 8 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, or 8 carbon atoms, such as ethenyl, 1-propenyl, 2-propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl, isobutenyl, sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, hexenyl, pentenyl, and octenyl. Preferably, the cycloalkenyl ring contains 3 to 8, i.e., 3, 4, 5, 6, 7 or 8 carbon atoms, such as 1-cyclopropenyl, 2-cyclopropenyl, 1-cyclobutenyl, 2-cyclobutenyl, 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, cyclohexenyl, cyclopentenyl, cyclooctenyl.

[0099] The term "alkynyl" refers to an unsaturated carbon atom containing chain or ring with one or more triple bonds. An example is the propargyl radical. Preferably, the alkynyl chain contains 2 to 8 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, or 8 carbon atoms, such as ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, pentynyl, and octynyl.

[0100] In one embodiment, carbon or hydrogen atoms in the alkyl, heteroalkyl, cycloalkyl, aryl, aralkyl, alkenyl, cycloalkenyl, and alkynyl radicals may be substituted, independently of one another, with one or more elements selected from the group consisting of O, S, and N, or with a group containing one or more elements selected from the group consisting of O, S, and N.

[0101] Embodiments include alkoxy, cycloalkoxy, aryloxy, aralkoxy, alkenyloxy, cycloalkenyloxy, alkynyloxy, alkylthio, cycloalkylthio, arylthio, aralkylthio, alkenylthio, cycloalkenylthio, alkynylthio, alkylamino, cycloalkylamino, arylamino, aralkylamino, alkenylamino, cycloalkenylamino, alkynylamino radicals.

[0102] Other embodiments include hydroxyalkyl, hydroxycycloalkyl, hydroxyaryl, hydroxyaralkyl, hydroxyalkenyl, hydroxycycloalkenyl, hydroxyalinyl, mercaptoalkyl, mercaptocycloalkyl, mercaptoaryl, mercaptoaralkyl, mercaptoalkenyl, mercaptocycloalkenyl, mercaptoalkynyl, aminoalkyl, aminocycloalkyl, aminoaryl, aminoaralkyl, aminoalkenyl, aminocycloalkenyl, aminoalkynyl radicals.

[0103] In another embodiment, the hydrogen atoms in the alkyl, heteroalkyl, cycloalkyl, aryl, aralkyl, alkenyl, cycloalkenyl, and alkynyl radicals may be replaced, independently of one another, by one or more halogen atoms. One radical is the trifluoromethyl radical.

[0104] Where two or more radicals or two or more residues may be selected independently of each other, the term "independently" means that the radicals or residues may be the same or different.

[0105] As used herein, phrases defining a length range limitation, such as, for example, "1 to 6," refer to any integer from 1 to 6, i.e., 1, 2, 3, 4, 5, and 6. In other words, any range defined by two explicitly stated integers is meant to include and disclose any integers defining said limitation and any integers falling within said range.

[0106] As used herein, the term "halo" refers to a halogen radical selected from the group consisting of F, Br, I, and Cl. Preferably, the halogen is F.

[0107] The phrase "protecting group" as used herein means temporary substituents which protect a potentially reactive functional group from undesired chemical transformations.

[0108] The term "amino-protecting group" or "N-terminal protecting group" refers to a group intended to protect the α-N-terminus of an amino acid or peptide, or otherwise protect the amino group of an amino acid or peptide from undesired reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, Protective Groups in Organic Synthesis, (John Wiley & Sons, New York (1981)), which is incorporated herein by reference. Furthermore, protecting groups can be used as prodrugs that are easily cleaved in vivo, for example, by enzymatic hydrolysis, to release the biologically active parent.α-N protecting groups include lower alkanoyl groups such as formyl, acetyl ("Ac"), propionyl, pivaloyl, and t-butylacetyl; other acyl groups such as 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, and 4-nitrobenzoyl; sulfonyl groups such as benzenesulfonyl and p-toluenesulfonyl; benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-ethoxybenzyloxycarbonyl, and 2-nitro-4,5-dimethoxybenzyloxycarbonyl. Carbamate-forming groups include benzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyoxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl; arylalkyl groups such as benzyl, triphenylmethyl, benzyloxymethyl, and 9-fluorenylmethyloxycarbonyl (Fmoc); and silyl groups such as trimethylsilyl. Other examples include theyl, succinyl, methoxysuccinyl, suberoyl, adipyl, azelayl, dansyl, benzyloxycarbonyl, methoxyazelayl, methoxyadipyl, methoxysuberoyl, and 2,4-dinitrophenyl. As used herein, the term "linker" refers to any chemically suitable linker.Preferably, the linker is not cleaved or is cleaved only slowly under physiological conditions.Therefore, preferably, the linker does not contain a recognition sequence for protease or other degradative enzymes.Because the compound of the present invention is preferably administered systemically, allowing broad access to all compartments of the body, and subsequently, the compound of the present invention is preferably concentrated wherever tumors are located in the body, it is preferable that the linker is selected so that it is not cleaved or is cleaved only slowly in blood.If less than 50% of the linker is cleaved 2 hours after the compound is administered to a human patient, cleavage is considered to be slow. Suitable linkers include, for example, optionally substituted alkyl, heteroalkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, sulfonyl, amine, ether, thioether, phosphine, phosphoramidate, carboxamide, ester, imidoester, amidine, thioester, sulfonamide, 3-thiopyrrolidine-2,5-dione, carbamate, urea, guanidine, thiourea, disulfide, oxime, hydrazine, hydrazide, hydrazone, diaza bond, triazole, triazoline, tetrazine, platinum complex, and amino acid, or a combination thereof. Preferably, the linker includes or consists of 1,4-piperazine, 1,3-propane, and phenol ether, or a combination thereof.

[0109] The term "optionally substituted" refers to a group in which one, two, three or more hydrogen atoms may be replaced, independently of one another, by respective substituents.

[0110] As used herein, the term "amino acid" refers to any organic acid containing one or more amino substituents, for example, an α-, β-, or γ-amino derivative of an aliphatic carboxylic acid.

[0111] The term "conventional amino acids" refers to the 20 naturally occurring amino acids and includes all of their stereoisomers, i.e., D, L-, D- and L-amino acids.

[0112] As used herein, the term "N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle" refers to a cyclic saturated or unsaturated hydrocarbon compound containing at least one nitrogen atom as a component of the cyclic chain.

[0113] Illustrative examples of pharmaceutically acceptable salts include, but are not limited to, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, clavulanate, cyclopentanepropionate, digluconate, dihydrochloride, dodecyl sulfate, edetate, edisylate, estolate, ethanesulfonate, ethanesulfonate, formate, fumarate, gluceptate, glucoheptonate, gluconate, glutamate, glycerophosphate, glycolylarsanilate, hemisulfate, heptanoate, hexanoate, and hexylresorcinate. (hexylresorcinate), hydrabamine, hydrobromide, hydrochloride, hydroiodide, 2-hydroxyethanesulfonic acid, hydroxynaphthoate, iodide, isothiocyanate, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methyl sulfate, mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate, N-methylglucamine Ammonium salts, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate / diphosphate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, undecanoate, valerate, etc. (See, e.g., Berge, SM, et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19.) Certain specific compounds of the present invention contain both basic and acidic functional groups that allow the compounds to be converted into either base or acid addition salts.

[0114] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but the salts are otherwise equivalent to the parent form of the compound for purposes of the present invention.

[0115] In addition to salt forms, the present invention provides compounds in prodrug form. Prodrugs of the compounds described herein are compounds that readily undergo chemical changes under physiological conditions to provide the compounds of Formula (I). Prodrugs are active or inactive compounds that are chemically modified into compounds of the present invention through in vivo physiological action, such as hydrolysis or metabolism, after administration of the prodrug to a patient. Furthermore, prodrugs can be converted to compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme. The properties and techniques involved in making and using prodrugs are well known to those skilled in the art. For a general discussion of prodrugs, including esters, see Svensson and Tunek, Drug Metabolism Reviews 16.5 (1988) and Bundgaard, Design of Prodrugs, Elsevier (1985).

[0116] Examples of masked carboxylate anions include various esters, such as alkyl (e.g., methyl, ethyl), cycloalkyl (e.g., cyclohexyl), aralkyl (e.g., benzyl, p-methoxybenzyl), and alkylcarbonyloxyalkyl (e.g., pivaloyloxymethyl). Amines have been masked as arylcarbonyloxymethyl-substituted derivatives, which are cleaved in vivo by esterases to release the free drug and formaldehyde (Bungaard J. Med. Chem. 2503 (1989)). Drugs containing an acidic NH group, such as imidazole, imide, and indole, have also been masked with N-acyloxymethyl groups (Bundgaard Design of Prodrugs, Elsevier (1985)).

[0117] Hydroxyl groups are masked as esters and ethers. EP 00039051 (Sloan and Little, April 11, 1981) discloses Mannich-base hydroxamic acid prodrugs, their preparation and use.

[0118] Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to the unsolvated forms and are intended to be encompassed within the scope of the present invention.

[0119] Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0120] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, geometric isomers, and individual isomers are all intended to be encompassed within the scope of the present invention.

[0121] The compounds of the present invention contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds, but still contain less than 100% of the molecules containing radioisotopic versions of that atom.

[0122] The term "pharmaceutical composition" as used herein refers to a substance and / or combination of substances used to identify, prevent, or treat a tissue condition or disease. A pharmaceutical composition is formulated to be suitable for administration to a patient to prevent and / or treat a disease. Furthermore, a pharmaceutical composition refers to a combination of an active agent and an inert or active carrier that makes the composition suitable for therapeutic use. Depending on its chemical and physical properties, a pharmaceutical composition can be formulated for oral, parenteral, topical, inhalation, rectal, sublingual, transdermal, subcutaneous, or vaginal administration. Pharmaceutical compositions include solids, semisolids, liquids, and transdermal therapeutic systems (TTS). Solid compositions are selected from the group consisting of tablets, coated tablets, powders, granules, pellets, capsules, effervescent tablets, or transdermal therapeutic systems. Liquid compositions are also included, selected from the group consisting of solutions, syrups, infusions, extracts, solutions for intravenous application, solutions for infusion, or solutions of the carrier systems of the present invention. Semisolid compositions that can be used in the context of the present invention include emulsions, suspensions, creams, lotions, gels, globules, buccal tablets, and suppositories.

[0123] "Pharmaceutically acceptable" means approved by a regulatory agency of the federal or state government or listed in the United States Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0124] The term "carrier," as used herein, refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic agent is administered. Such pharmaceutical carriers can be sterile liquids, such as saline and oils (including those of petroleum, animal, vegetable, or synthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesame oil, etc.). Saline is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline and aqueous dextrose and glycerol solutions can also be used as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. If desired, the composition may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin.

[0125] III. Structural overview of FAP-activated radiopharmaceuticals In some embodiments, the FAP-activated theranostic prodrug is [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, R represents a ligand-targeting theranostic moiety that includes a ligand for binding to a cellular target and one or more of a radioactive moiety and / or a chelator for chelating the radioactive moiety; A represents a 5- to 8-membered heterocycle; X is O or S; R 10 is an amino terminal blocking group; R 12 is hydrogen or (C1-C6) alkyl; R 13is hydrogen, (C1-C6) alkyl (which may be linear or branched) or (C1-C6); R 14 is, independently for each occurrence, -(C1-C6)alkyl, -OH, -NH2, or halogen; p is an integer from 0 to 6; L is a bond or a self-eliminating linker that upon cleavage by FAP releases NH2-LR; Enzymatic cleavage of the prodrug by fibroblast activation protein (FAP) results in release of the ligand-targeted theranostic moiety either as an activated ligand-targeted theranostic moiety (i.e., its pharmacologically active form) or in a form that is readily metabolized to that active form; and, upon release from the prodrug by FAP cleavage, the activated ligand-targeted theranostic moiety binds to the cellular target with a Kd for binding to the cellular target that is less than the Kd for binding of the prodrug to the cellular target (i.e., has a higher affinity for the cellular target).

[0126] In certain preferred embodiments of Formula II, R 12 is H.

[0127] In certain embodiments of the structure of formula II, R 10 forms an amide or thioamide with the nitrogen to which it is attached, and the prodrug has the formula: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, A, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0128] In certain preferred embodiments of Formula IIa, X is O; and / or R 12 is H. In certain preferred embodiments, X is O and R 12 is H.

[0129] In certain preferred embodiments, X is O and R 11 is -(C1~C 10 )Alkyl-CO2H, -(C1-C 10 ) alkyl-CO2H, or -(C1-C 10 )aryl-CO2H.

[0130] In certain preferred embodiments, X is O and —C(═O)—R 11 forms an acyl group of the carboxylic acid, such as, for example, a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0131] In a preferred embodiment, X is O and R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, and m is an integer from 1 to 6. In certain embodiments, m is 1 or 2. In certain embodiments, R 11a represents a 6-membered aryl or heteroaryl group.

[0132] In certain embodiments, the FAP-activated theranostic prodrug is [ka] or a pharmaceutically acceptable salt thereof, wherein R, R 10 , R 12 , R 13 , R 14 , L, X and p are as defined above for Formula II.

[0133] In certain preferred embodiments of Formula III, R 12 is H.

[0134] In certain embodiments of the structure of formula III, R 10 forms an amide or thioamide with the nitrogen to which it is attached, and the prodrug has the formula IIIa: [ka] or a pharmaceutically acceptable salt thereof; During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0135] In certain preferred embodiments of Formula IIIa, X is O; and / or R 12 is H. In certain preferred embodiments, X is O and R 12 is H.

[0136] In certain preferred embodiments, X is O and R 11 Ha-(C1~C10 )Alkyl-CO2H, -(C1-C 10 ) alkenyl-CO2H, or -(C1-C 10 )aryl-CO2H.

[0137] In certain preferred embodiments, X is O and —C(═O)—R 11 forms an acyl group of the carboxylic acid, for example, by way of example, a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0138] In a preferred embodiment, X is O and R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, and m is an integer from 1 to 6. In certain embodiments, m is 1 or 2. In certain embodiments, R 11a represents a 6-membered aryl or heteroaryl group.

[0139] In certain embodiments, the FAP-activated theranostic prodrug has Formula IV: [ka] or a pharmaceutically acceptable salt thereof, wherein R 13 is (C1-C6) alkyl (which may be linear or branched) or (C1-C6), and R, R 10 , R 12 , R 13 , R 14 , L, X and p are as defined above for formula II.

[0140] In certain preferred embodiments of Formula IV, R 13 is methyl; p is zero; and / or R12 is H. In certain preferred embodiments, R 13 is methyl and p is zero (i.e., R 14 does not exist), R 12 is H

[0141] In certain embodiments of the structure of formula IV, R 10 forms an amide or thioamide together with the nitrogen to which it is attached, and the prodrug is of formula IVa [ka] or a pharmaceutically acceptable salt thereof, During the ceremony, X is O or S; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R, R 12 , R 13 , R 14 , p and L are as described for formula II.

[0142] In certain preferred embodiments of Formula IVa, R 13 is methyl; X is O; p is zero; and / or R 12 is H. In certain preferred embodiments, R 13 is methyl, X is O, p is zero, and R 12 is H.

[0143] In certain preferred embodiments, X is O and R 11 Ha-(C1~C 10 )Alkyl-CO2H, -(C1-C 10 ) alkenyl-CO2H, or -(C1-C 10)aryl-CO2H.

[0144] In certain preferred embodiments, X is O and —C(═O)—R 11 forms an acyl group of the carboxylic acid, for example, by way of example, a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0145] In a preferred embodiment, X is O and R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, and m is an integer from 1 to 6. In certain embodiments, m is 1 or 2. In certain embodiments, R 11a represents a 6-membered aryl or heteroaryl group.

[0146] In certain embodiments, the FAP-activated radiopharmaceutical is [ka] is represented by During the ceremony, R, R 10 , R 13 and L is as described for formula II; R 11 -(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5; R 13 is (C1-C6) alkyl (which may be straight or branched chain) or (C1-C6).

[0147] In certain preferred embodiments of Formulas V and Va, R 13 is methyl.

[0148] In certain preferred embodiments, X is O and R 11 Ha-(C1~C 10 )Alkyl-CO2H, -(C1-C 10 ) alkenyl-CO2H, or -(C1-C 10 )aryl-CO2H.

[0149] In certain preferred embodiments, X is O and —C(═O)—R 11 forms an acyl group of the carboxylic acid, for example, by way of example, a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0150] In a preferred embodiment, X is O and R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, and m is an integer from 1 to 6. In certain embodiments, m is 1 or 2. In certain embodiments, R 11a is a 6-membered aryl or heteroaryl group.

[0151] In certain embodiments, the FAP-activated theranostic prodrug has Formula VI: [ka] is represented by During the ceremony, R, R 10 and L is as described for formula II; R 11-(C=X) taken together represent an acyl N-terminal blocking group; or R 11 is -(C1~C 10 ) alkyl, -(C1-C 10 ) alkoxy, -(C1-C 10 ) alkyl-C(O)-OH, -(C1-C 10 ) alkenyl-C(O)-OH, -(C1-C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(C3-C8) cycloalkyl, -(C3-C8) cycloalkyl(C1-C 10 ) alkyl, -(C6-C 14 ) aryl, -aryl (C1-C 10 ) alkyl, -O-(C1-C4) alkyl-(C6-C 14 ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C1-C 10 ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C1~C 10 ) alkyl, -(AA) n -(C1~C 10 ) alkoxy, -(AA) n -(C1~C 10 )Alkyl-C(O)-(C1-C 10 ) alkyl, -(AA) n -(C3-C8)cycloalkyl, -(AA) n -(C3-C8)cycloalkyl(C1-C 10 ) alkyl, -(AA) n -(C6~C 14 ) aryl, -(AA) n -Aryl (C1-C 10 ) alkyl, -(AA) n -5 to 10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl (C1-C 10) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5;

[0152] In certain preferred embodiments, X is O and R 11 is -(C1~C 10 )Alkyl-CO2H, -(C1-C 10 ) alkenyl-CO2H, or -(C1-C 10 )aryl-CO2H.

[0153] In certain preferred embodiments, X is O and —C(═O)—R 11 forms an acyl group of the carboxylic acid, for example, by way of example, a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl, or methoxysuccinyl group.

[0154] In a preferred embodiment, X is O and R 11 Ha-(CH2) m -R 11a where R 11a is a 5-10 membered aryl or heteroaryl group, and m is an integer from 1 to 6. In certain embodiments, m is 1 or 2. In certain embodiments, R 11a is a 6-membered aryl or heteroaryl group.

[0155] In yet other embodiments, the FAP-activated theranostic prodrug has Formula VII: [ka] wherein R and L are as defined for formula II above, and -C(=O)R 11 forms an acyl group.

[0156] In certain embodiments of structures II-VII above, —C(═X)R 11 or -C(=O)R 11 forms an acyl group.

[0157] In certain embodiments, the acyl group is selected from the group consisting of aryl(C1-C6)acyl and heteroaryl(C1-C6)acyl.

[0158] In certain embodiments, the aryl(C1-C6)acyl is a (C1-C6)acyl substituted with an aryl selected from the group consisting of benzyl, naphthalenyl, phenanthrenyl, phenolyl, and anilinyl.

[0159] In certain embodiments, the aryl(C1-C6)acyl is a (C1)acyl substituted with an aryl selected from the group consisting of benzyl, naphthalenyl, phenanthrenyl, phenolyl, and anilinyl.

[0160] In certain embodiments, the acyl group is heteroaryl(C1-C6)acyl.

[0161] In certain embodiments, the heteroaryl(C1-C6)acyl is a (C1-C6)acyl substituted with a heteroaryl selected from the group consisting of pyrryl, furyl, thiophenyl (a / k / a thienyl), imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl.

[0162] In certain embodiments, the heteroaryl(C1-C6)acyl is a (C1)acyl substituted with a heteroaryl selected from the group consisting of pyrryl, furyl, thiophenyl (a / k / a thienyl), imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl.

[0163] In certain embodiments, the FAP substrate portion comprises (d)-Ala (or other (d)-amino acid at that position formed by R3) at the third amino acid position, optionally at the N-terminus, and optionally the amino acid at the third amino acid position is serine or threonine.

[0164] a. Self-Eliminating Linker In certain embodiments, the FAP substrate moiety is linked to the ligand-targeted theranostic moiety via a self-elimination linker (L in the formula above). Cleavage of the FAP substrate moiety by FAPα then releases the activated ligand-targeted theranostic moiety by removal of the self-elimination linker.

[0165] A self-eliminating moiety can be defined as a bifunctional chemical group capable of covalently linking two separate chemical moieties into a generally stable molecule, one of which can be released from the molecule by enzymatic cleavage; and following enzymatic cleavage, which can spontaneously cleave from the remainder of the bifunctional chemical group, releasing the other of the separate chemical moieties. Thus, in some embodiments, the self-eliminating moiety is covalently attached at one of its ends to a ligand via an amide bond, either directly or indirectly through a spacer unit, and at the other end to a chemically reactive site (functional group) attached to a therapeutic moiety.

[0166] Therapeutic conjugates should typically be stable in circulation, or at least stable in the absence of an enzyme capable of cleaving the amide bond between the substrate recognition sequence (FAPα-cleavable linker) and the self-elimination moiety. Exposure of the therapeutic conjugate to an appropriate enzyme (FAPα) cleaves the amide bond, initiating a spontaneous self-elimination reaction that results in cleavage of the bond covalently linking the self-elimination moiety to the therapeutic moiety, thereby releasing the free therapeutic moiety in its underivatized or pharmacologically active form. The self-elimination moiety in the conjugate incorporates one or more heteroatoms, thereby improving solubility, improving cleavage rate, and reducing the tendency of the conjugate to aggregate.

[0167] In some embodiments, L is a benzyloxycarbonyl group. In other embodiments, the self-eliminating linker L is -NH-(CH)-C(=O)- or -NH-(CH)-C(=O)-. In yet other embodiments, the self-eliminating linker L is p-aminobenzyloxycarbonyl (PABC). In yet other embodiments, the self-eliminating linker L is 2,4-bis(hydroxymethyl)aniline.

[0168] The therapeutic conjugates of the present disclosure can employ a heterocyclic self-elimination moiety covalently linked to a therapeutic moiety and a cleavable substrate recognition sequence. A self-elimination moiety can be defined as a bifunctional chemical group capable of covalently linking two separate chemical moieties into a generally stable molecule, one of which can be released from the molecule by enzymatic cleavage; and, following enzymatic cleavage, spontaneously cleaves away from the remainder of the bifunctional chemical group, releasing the other separate chemical moiety. According to the present disclosure, the self-elimination moiety can be covalently attached to a ligand at one end via an amide bond, either directly or indirectly via a spacer unit, and covalently attached to a chemical reactive site (functional group) on a drug at the other end. Derivatization of a therapeutic moiety with a self-elimination moiety can render the drug less pharmacologically active (e.g., less toxic) or completely inactive until the drug is cleaved.

[0169] Therapeutic conjugates should typically be stable in circulation, or at least stable in the absence of enzymes capable of cleaving the amide bond between the substrate recognition sequence and the self-elimination moiety. However, exposure of the therapeutic conjugate to an appropriate enzyme cleaves the amide bond, initiating a spontaneous self-elimination reaction that results in cleavage of the bond covalently linking the self-elimination moiety to the drug, thereby releasing the free therapeutic moiety in its underivatized or pharmacologically active form.

[0170] The self-eliminating moieties in the conjugates of the present disclosure, in some embodiments, incorporate one or more heteroatoms, thereby providing improved solubility, improved cleavage rates, and reduced tendency of the conjugates to aggregate. These improvements of the heterocyclic self-eliminating linker constructs of the present disclosure over non-heterocyclic PAB-type linkers can result in surprising and unexpected biological properties, such as increased potency, reduced toxicity, and more favorable pharmacokinetics.

[0171] In some embodiments, L is a benzyloxycarbonyl group.

[0172] In some embodiments, L is [ka] where R 1 is hydrogen, unsubstituted or substituted C 1-3 alkyl, or unsubstituted or substituted heterocyclyl. In some embodiments, R 1 is hydrogen. In some instances, R 1 is methyl.

[0173] In some embodiments, L is [ka] is selected from.

[0174] In some embodiments, the self-elimination moiety, L, is [ka] is selected from During the ceremony, U is O, S or NR 6 and; Q is CR 4 or N; V 1 , V 2 and V 3 is an independent CR 4 or N, with the proviso that for Formulas II and III, Q, V 1 and V 2 at least one of is N; T is NH, NR attached to the therapeutic moiety 6 , O or S; R 1 , R 2 , R 3 and R 4 are independently H, F, Cl, Br, I, OH, -N(R 5 )2, -N(R 5 )3 +, C1-C8 alkyl halides, carboxylates, sulfates, sulfamates, sulfonates, -SO2R 5 , -S(=O)R 5 , -SR 5 , -SO2N(R 5 )2, -C(=O)R 5 , -CO2R 5 , -C(=O)N(R 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 halosubstituted alkyl, polyethyleneoxy, phosphonate, phosphate, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocycles and C1-C 20 substituted heterocycles; or R 2 and R 3 together form a carbonyl (=O) or a spirocarbocyclic ring of 3 to 7 carbon atoms; R 5 and R 6 are independently H, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocycles and C1-C 20 substituted heterocycles; where C1 to C8 substituted alkyl, C2 to C8 substituted alkenyl, C2 to C8 substituted alkynyl, C6 to C 20 Substituted aryl and C2-C 20 The substituted heterocycles are independently selected from F, Cl, Br, I, OH, -N(R 5 )2, -N(R 5 )3 + , C1-C8 alkyl halide , carboxylate, sulfate, sulfamate, sulfonate, C1-C8 alkyl sulfonate, C1-C8 alkyl amino, 4-dialkylaminopyridinium, C1-C8 alkyl hydroxyl, C1-C8 alkyl thiol, -SO2R 5 , -S(=O)R 5 , -SR 5 , -SO2N(R 5 )2, -C(=O)R 5 , -CO2R 5 , -C(=O)N(R 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 trifluoroalkyl, C1-C8 alkyl, C3-C 12 Carbocycle, C6~C 20 Aryl, C2-C 20 It is substituted with one or more substituents selected from heterocycle, polyethyleneoxy, phosphonate, and phosphate.

[0175] It will be understood that when T is NH, it is derived from a primary amine (-NH) attached to the therapeutic moiety (prior to coupling to the self-eliminating moiety), and when T is N, it is derived from a secondary amine (-NH) attached to the therapeutic moiety (prior to coupling to the self-eliminating moiety). Similarly, when T is O or S, it is derived from a hydroxyl (-OH) or sulfhydryl (-SH) group, respectively, attached to the therapeutic moiety prior to coupling to the self-eliminating moiety.

[0176] In some embodiments, the self-elimination linker L is —NH—(CH 2 ) 4 —C(═O)— or —NH—(CH 2 ) 3 —C(═O)—.

[0177] In some embodiments, the self-elimination linker L is p-aminobenzyloxycarbonyl (PABC).

[0178] In some embodiments, the self-elimination linker L is 2,4-bis(hydroxymethyl)aniline.

[0179] Other examples of self-eliminating linkers readily adapted for use in the therapeutic conjugates described herein are taught, for example, in U.S. Pat. No. 7,754,681; WO 2012 / 074693 A1; U.S. Pat. No. 9,089,614; European Patent Application Publication No. 1,732,607; WO 2015 / 038426 A1 (all of which are incorporated by reference); Walther et al. "Prodrugs in medicinal chemistry and enzyme prodrug therapies" Adv Drug Deliv Rev. 2017 Sep 1; 118:65-77; and Tranoy-Opalinski et al. "Design of self-eliminating linkers for tumor-activated prodrug therapy", Anticancer Agents Med Chem. 2008 Aug;8(6):618-37 (the teachings of each are incorporated by reference herein).

[0180] Still other non-limiting examples of self-elimination linkers for use in accordance with the present disclosure are described in International Publication No. WO2019 / 236567, published December 12, 2019, which is incorporated herein by reference.

[0181] b. Targeting moiety In certain embodiments, the ligand-targeting theranostic moiety (R) is -TM-L 1 -R 20 is represented by During the ceremony, TM represents a ligand targeting moiety that selectively binds to a cell surface feature on a target cell; L 1 represents a bond or a linker; R 20 represents a radioactive moiety, a chelator, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, an imaging agent, or a detectable nanoparticle.

[0182] For example, the ligand targeting moiety TM can be a moiety that selectively binds to a cell surface feature on tumor cells or tumor stromal cells.

[0183] For example, the ligand targeting moiety TM can be a moiety that selectively binds to a protein, carbohydrate, or lipid (such as a glycolipid) on a target cell.

[0184] In certain embodiments, the cell surface feature causes the ligand-targeted theranostic moiety to be internalized upon binding of the ligand-targeted theranostic moiety to the cell surface feature.

[0185] In certain embodiments, the ligand targeting moiety selectively binds to a protein on the target cell, hi certain embodiments, the protein on the target cell is a receptor.

[0186] In certain embodiments, the receptor is a G protein-coupled receptor (GPCR), such as a gastrin-releasing peptide receptor (e.g., a bombesin receptor such as BB1, BB2, or BB3), a calcitonin receptor, an oxytocin receptor, a somatostatin receptor (e.g., somatostatin receptor subtype 2), a melanocortin receptor (e.g., MC1R), a cholecystokinin receptor (e.g., a cholecystokinin B receptor), a neurotensin receptor, or a neuropeptide (or neuropeptide) Y receptor.

[0187] In other embodiments, the receptor is a growth factor receptor, such as an epidermal growth factor receptor (e.g., ErbB1, ErbB2, ErbB3, or ErbB4), an insulin growth factor receptor (e.g., IGFR1 or IGFR2), a TGFβ receptor (e.g., TGFβR1 or TGFβR2), a VEGF receptor (e.g., VEGFR1, VEGFR2, VEGFR3, or VEGFR4), a PDGF receptor (e.g., PDGFRα or PDGFRβ), or an FGF receptor (e.g., FGFR1, FGFR2, FGFR3, or FGFR4).

[0188] In certain embodiments, the receptor binding moiety binds to the folate receptor alpha and can be a folate receptor ligand, such as folic acid or a folic acid analog (e.g., etalfolatide, vintafolide, leucovorin, and methotrexate).

[0189] In certain instances, the ligand targeting moiety can be selected to bind to an integrin. In certain embodiments, the ligand targeting moiety binds to integrin α v Binds to β3.

[0190] In certain embodiments, the ligand targeting moiety may be selected to bind to an N-acetyl-L-aspartyl-L-glutamic peptidase, such as prostate-specific membrane antigen (PSMA).

[0191] The ligand targeting moiety may itself have pharmacological activity of its own, or may be inert and simply serve the purpose of delivering the ligand-targeting theranostic moiety to cells (preferably intracellularly) that express the receptor.

[0192] In certain embodiments, the ligand targeting moiety is somatostatin or a somatostatin analog, such as octreotate, octreotide, or pentetreotide.

[0193] In certain embodiments, the ligand targeting moiety binds to αIIbβ3 and may be an αIIbβ3 targeting ligand such as RGD or an RGD analog (i.e., a dimeric or multimeric analog), including exemplary cyclic RGD peptides such as cyclo(-Arg-Gly-Asp-D-Phe Val-)["c(RGDfV)"], c(RGDfK), c(RGDfC), c(RADfC), c(RADfK), c(RGDfE), c(RADfE), RGDSK, RADSK, RGDS, c(RGDyC), c(RADyC), c(RGDyE), c(RGDyK), c(RADyK), and HE[c(RGDyK)]2, EMD12194, DMP728, DMP757, and SK&F107260.

[0194] To further illustrate, in certain embodiments, the ligand-targeting moiety binds to prostate-specific membrane antigen (PSMA). For example, the ligand-targeting theranostic moiety (R) in the above structure is: [ka] can be expressed as During the ceremony, L 1 represents a bond or a linker; R 20 represents a radioactive moiety, a chelator, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, an imaging agent, or a detectable nanoparticle; R 30 represents independently for each occurrence hydrogen or lower alkyl.

[0195] In certain embodiments, L 1 represents a linker. In certain embodiments, the linker L 1 is selected to provide some hydrophobic contact with PSMA. In certain embodiments, the linker L 1 is selected to provide some hydrophobic contact with PSMA.

[0196] In certain embodiments, -L 1 -R 20 teeth, [ka] where R 20 is as defined above, and R 31 is -(CH2) p -aryl or -(CH2) p -heteroaryl, where p is 0, 1, 2, 3, or 4. In certain embodiments, p is 1 or 2, preferably p is 1. In certain embodiments, R 31 is -CH2-aryl, where the aryl group is C6-C 12aryl and is a monocyclic ring or a bicyclic fused ring. In certain preferred embodiments, R 31 is -CH2-naphthalene.

[0197] In certain embodiments, R 20 is a chelator (or chelating agent) and may contain a chelated radioisotope.

[0198] In certain embodiments, -L1-R 20 teeth, [ka] or a radioisotope chelated formulation thereof.

[0199] In certain embodiments, R 20 is F 18 For illustration, -L 1 -R 20 teeth, [ka] may be selected from:

[0200] By way of further example, in certain embodiments, the ligand-targeting moiety binds to a folate receptor. For example, the ligand-targeting theranostic moiety (R) comprises folic acid or a folic acid analog, such as [ka] can be expressed as one of During the ceremony, R 21 represents H, and R 22 is -NH-(CH2) q -R 20 , -NH-(CH2) q -NH-C(O)-(CH2) q -R 20 or -NH-(CH2) q -C(O)-(CH2) q -R 20 represents; or R 22 represents H, and R 21 is -NH-(CH2) q -R 20 or -NH-(CH2) q -C(O)-(CH2) q -R 20 represents; or R 21 Or R 22 One of them represents H and the other represents [ka] selected from the group consisting of: R 23 represents H, -CH3, -CH2CH3, or -CO2H; R 20 represents a radioactive moiety, a chelator, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, an imaging agent, or a detectable nanoparticle; q is independently 0, 1, 2, 3, or 4 for each occurrence.

[0201] In certain embodiments, R 20 represents the chelating moiety, and R 21 -NH-CH2-R 20 , -NH-CH2-C(O)-R 20 , -NH-C(O)-CH2-R 20 , -NH-CH2-C(O)-CH2-R 20 or -NH-(CH2)2-NH-C(O)-CH2-R 20 represents R 22 represents H.

[0202] In certain embodiments, R 20 represents the chelating moiety, and R 21 represents H, and R 22 is -NH-CH2-R 20 , -NH-CH2-C(O)-R 20 , -NH-C(O)-CH2-R 20 , -NH-CH2-C(O)-CH2-R 20 , or -NH-(CH2)2-NH-C(O)-CH2-R20 Represents.

[0203] For example, the ligand-targeting theranostic moiety (R) can be: [ka] It could be.

[0204] In yet other embodiments, the ligand-targeting theranostic moiety (R) comprises a directly radiolabeled folic acid or folic acid analog, such as: [ka] In the formula, R 23 represents H, -CH3, -CH2CH3 or -CO2H, and X represents CR 40 or N, R 40 is H or lower alkyl.

[0205] In certain embodiments, the ligand targeting moiety binds to a somatostatin receptor (such as somatostatin receptor subtype 2). For example, the ligand targeting theranostic moiety (R) can comprise somatostatin or a somatostatin analog. Examples of somatostatin analogs include octreotide, octreotate, lanreotide, vapreotide, pasireotide, seglitide, venereotide, KE-108, SDZ-222-100, Sst3-ODN-8, CYN-154806, JR11, J2156, SRA-880, ACQ090, P829, SSTp-58, SSTp-86, BASS, and somatoprim.

[0206] In certain embodiments, the somatostatin analog is a somatostatin receptor agonist.

[0207] For example, the ligand-targeting theranostic moiety (R) can be: [ka] [ka] and During the ceremony, L 1 represents a bond or a linker; R 20 represents a radioactive moiety, a chelator, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, an imaging agent, or a detectable nanoparticle.

[0208] In certain embodiments, R 20 is a chelating moiety. By way of example, R may be DOTA-octreotate, e.g., [ka] is.

[0209] Also, R is (DOTA 0 -Phe 1 -Tyr 3 ) octreotide, for example: [ka] is.

[0210] In certain embodiments, R 20 teeth, 18 It is the moiety containing the F group.

[0211] R is NOTA-octreotide (shown below [ 18 F]AlF-NOTA-octreotide), for example: [ka] is.

[0212] or, 18 F may be a direct substituent on the somatostatin or somatostatin analog, or R 20 -L 1 -but [ka] It may also be part of a non-chelating tracer moiety, such as when:

[0213] In still other embodiments, the ligand targeting moiety can be selected from a bombesin analog, a calcitonin analog, an oxytocin analog, an EGF analog, an α-melanocyte stimulating hormone analog, a minigastrin analog, a neurotesin analog, and a neuropeptide Y (NPY) analog.

[0214] c. Radioisotopes, chelators, and other theranostic labels In certain embodiments, the ligand (R in Formulas I, II, and III) comprises a radioactive moiety, wherein the radioactive moiety comprises a fluorescent isotope, a radioisotope, a radiopharmaceutical, or a combination thereof. Preferably, the radioactive moiety comprises a radioisotope selected from the group consisting of an alpha-emitting isotope, a beta-emitting isotope, a gamma-emitting isotope, an Auger electron-emitting isotope, an X-ray-emitting isotope, and a fluorescent-emitting isotope.

[0215] The radioisotope can be selected to allow for imaging and / or radiotherapy.

[0216] The radioisotope may include a radioactive metal or metalloid isotope. Preferably, the radioisotope is a water-soluble metal cation.

[0217] Exemplary radioisotopes include: 18 F, 43 K. 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 75 Br,76 Br, 77 As, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99m Tc, 100 Pd, 101m Rh, 103 Pb, 105 Rh, 109 Pd, 111 Ag, 111 In, 113 In, 119 Sb 121 Sn, 123 I, 124 I, 125 I, 127 Cs, 128 Ba, 129 Cs, 131 Cs, 131 I, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 Eu, 153 Eu, 153 Sm, 159 Gr, 161 Tb, 165 Dy, 166 Ho, 169 Eu, 175 Yb, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, and 227 Th are included.

[0218] In certain embodiments, the radioisotope is intended to enable imaging, such as by SPECT imaging and / or PET imaging. Single-photon emission computed tomography (SPECT) is a nuclear medicine tomography technique that uses gamma rays and can provide true 3D information. The information is often presented as cross-sectional slices across the patient. The gamma-ray emission of the isotope allows for the location of the radiolabeled substance's accumulation within the patient's body to be seen. Such true 3D representations can be useful in tumor imaging. Positron emission tomography (PET) is a nuclear medicine imaging technique that produces 3D images and has greater sensitivity than traditional SPECT imaging. The system detects gamma-ray pairs indirectly emitted by a positron-emitting radionuclide (tracer) introduced into the body. A 3D image of the tracer concentration within the body is then constructed by computer analysis, often achieved with the aid of a computed tomography (CT) X-ray scan performed on the patient during the same session on the same machine. Positron-emitting isotopes can also be used in conjunction with CT to provide 3D imaging of the anatomical distribution of labeled medical devices.

[0219] In certain embodiments, the radioisotope is an element of Group XIII (boron family) of the periodic table, which includes Ga and In. Particularly preferred radioisotopes include Ga-67, Ga-68, Lu-177, Y-90, and In-111. Most preferably, the radioisotopes are Lu-177 and Y-90. In one embodiment, the radioisotope is Lu-177.

[0220] In certain embodiments, the radioisotope is a transition metal such as Lu-177, Y-90, Cu-64, Cu-67, and Tb-161. Preferably, the radioisotope is Lu-177 or Y-90.

[0221] In certain embodiments, the ligand may comprise a combination of at least two radioisotopes to enable imaging and / or therapy. The combination of radioisotopes may be selected from Ga-68 and Lu-177; Ga-67 and Y-90; Ga-68 and Y-90; In-111 and Y-90; Lu-177 and Y-90; and Ga-67 and Tb-161.

[0222] The present invention can further comprise the use of at least one non-radioactive and non-toxic carrier metal.For example, the carrier metal can be selected from Bi and Fe.For example, the non-radioactive carrier metal can be one that allows MRI imaging (for example, Fe) or X-ray contrast imaging (for example, Bi).Another example of a carrier metal includes trivalent bismuth, which can also provide X-ray contrast to the microsphere and can be imaged by CT.

[0223] In certain embodiments, the ligand comprises a chelating moiety, eg, a chelator for a radiometal or paramagnetic ion.

[0224] The chelating agent can include any chelator known in the art, see, e.g., Parus et al., "Chemistry and bifunctional chelating agents for binding (177)Lu," Curr Radiopharm. 2015; 8(2):86-94; Wangler et al., "Chelating agents and their use in radiopharmaceutical sciences," Mini Rev Med Chem. 2011 October; 11(11):968-83; Liu, "Bifunctional Coupling Agents for Radiolabeling of Biomolecules and Target-Specific Delivery of Metallic Radionuclides," Adv Drug Deliv Rev. 2008 September; 60(12): 1347-1370.

[0225] Specific examples include, for example: [ka] [ka] [ka] [ka] [ka]

[0226] Further illustrative examples include, for example: [ka]

[0227] In certain preferred embodiments, the ligand may comprise DOTA, ie, may be covalently attached to the ligand via any of its four carboxylic acid groups.

[0228] In certain embodiments, the chelator comprises a radioisotope chelated thereto.

[0229] In certain embodiments, the chelator comprises a paramagnetic ion chelated thereto, such as chromium(III), manganese(II), iron(III), iron(II), cobalt(II), nickel(II), copper(II), neodymium(III), samarium(III), ytterbium(III), gadolinium(III), vanadium(II), terbium(III), dysprosium(III), holmium(III), erbium(III), or a combination of these paramagnetic ions.

[0230] When the moiety is a detectable label, it can be a fluorescent label.That is, in certain embodiments, the ligand comprises a fluorescent dye conjugated to the ligand, such as xanthene, acridine, oxazine, cyanine, styryl dye, coumarin, porphine, metal-ligand complex, fluorescent protein, nanocrystal, perylene, boron-dipyrromethene, and phthalocyanine, and the conjugates and combinations of these types of dyes.Specific examples of fluorescent labels include, but are not limited to, organic dyes such as cyanine, fluorescein, rhodamine, Alexa Fluor, Dylight fluor, ATTO Dyes, BODIPY Dyes, and the like, as well as biological fluorophores such as green fluorescent protein (GFP), R-phycoerythrin, and quantum dots.

[0231] In certain embodiments, the fluorescent moiety is selected from the group consisting of Cy5, Cy5.5 (also known as Cy5++), Cy2, fluorescein isothiocyanate (FITC), tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin, Cy7, fluorescein (FAM), Cy3, Cy3.5 (also known as Cy3++), Texas Red, LightCycler-Red 640, LightCycler-Red 705, tetramethylrhodamine (TMR), rhodamine, rhodamine derivative (ROX), hexachlorofluorescein (HEX), rhodamine 6G (R6G), rhodamine derivative JA133, Alexa Fluorescent Dyes (e.g., Alexa Fluor 488, Alexa Fluor 546, Alexa Fluor 633, Alexa Fluor 555, and Alexa Fluor Fluorescent compounds may be selected from the group consisting of fluorescent transition metal complexes such as Fluorescent Protein (GFP), Fluorescent Protein (EBFP), Fluorescent Protein 2 (EBFP2), Fluorescent Protein 3 (EBFP2), Fluorescent Protein 4 (EBFP4), Fluorescent Protein 5 (EBFP5), Fluorescent Protein 6 (EBFP6), Fluorescent Protein 7 (EBFP7), Fluorescent Protein 8 (EBFP8), Fluorescent Protein 9 (EBFP9), Fluorescent Protein 10 (EBFP10), Fluorescent Protein 11 (EBFP11), Fluorescent Protein 12 (EBFP12), Fluorescent Protein 13 (EBFP14), Fluorescent Protein 14 (EBFP15), Fluorescent Protein 15 (EBFP16), Fluorescent Protein 16 (EBFP17), Fluorescent Protein 17 (EBFP18), Fluorescent Protein 18 (EBFP19), Fluorescent Protein 19 (EBFP21), Fluorescent Protein 19 (CFP19 ...

[0232] IV. Exemplary Therapeutic Uses of FAP-Activated Radiopharmaceuticals Yet another aspect of the present invention provides a method for diagnosing, imaging, or reducing tissues that overexpress a FAP in an animal (preferably a human patient), comprising administering to the animal a FAP-activated theranostic prodrug of the present invention.

[0233] In some embodiments, the tissue that overexpresses FAP is a tumor, particularly a solid tumor. In some embodiments, the tumor is selected from the group consisting of: colorectal tumor, pancreatic tumor, lung tumor, ovarian tumor, liver tumor, breast tumor, kidney tumor, prostate tumor, neuroendocrine tumor, gastrointestinal tumor, melanoma, cervical tumor, bladder tumor, glioblastoma, and head and neck tumor. In some embodiments, the tumor is a colorectal tumor. In some embodiments, the tumor is an ovarian tumor. In some embodiments, the tumor is a lung tumor. In some embodiments, the tumor is a pancreatic tumor. In some embodiments, the tumor is a melanoma. In some embodiments, the tumor is a bladder tumor.

[0234] By way of further example, the subject FAP-activated radiopharmaceutical prodrugs may be used to treat osteosarcoma, rhabdomyosarcoma, neuroblastoma, kidney cancer, leukemia, renal transitional cell carcinoma, bladder cancer, Wilms' carcinoma, and leukemia. It can be used to treat patients suffering from cancer such as ovarian cancer, pancreatic cancer, breast cancer (including triple-negative breast cancer), prostate cancer, bone cancer, lung cancer (e.g., small cell or non-small cell lung cancer), gastric cancer, colorectal cancer, cervical cancer, synovial sarcoma, head and neck cancer, squamous cell carcinoma, multiple myeloma, renal cell carcinoma, retinoblastoma, hepatocellular carcinoma, melanoma, rhabdoid tumor of the kidney, Ewing's sarcoma, chondrosarcoma, brain tumor, glioblastoma, meningioma, pituitary adenoma, vestibular schwannoma (schwannoma), primitive neuroectodermal tumor, medulloblastoma, astrocytoma, anaplastic astrocytoma, oligodendroglioma, ependymoma, choroid plexus papilloma, polycythemia vera, thrombocythemia, idiopathic myelofibrosis, soft tissue sarcoma, thyroid cancer, endometrial cancer, carcinoid cancer or liver cancer, breast cancer or gastric cancer. In some embodiments of the present disclosure, the cancer is a metastatic cancer, such as any of the various cancers described above.

[0235] In some embodiments, in addition to administering a FAP-activating radiopharmaceutical prodrug described herein, the method or treatment further comprises administering at least one additional immune response stimulator. In some embodiments, the additional immune response stimulator includes, but is not limited to, colony-stimulating factors (e.g., granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF)), interleukins (e.g., IL-1, IL2, IL-3, IL-7, IL-12, IL-15, IL-18), checkpoint inhibitors, antibodies that block immunosuppressive function (e.g., anti-CTLA-4 antibodies, anti-CD28 antibodies, anti-CD3 antibodies), Toll-like receptors (e.g., TLR4, TLR7, TLR9), or members of the B7 family (e.g., CD80, CD86). The additional immune response stimulator can be administered before, simultaneously with, and / or after administration of the FAP-activating radiopharmaceutical prodrug. Also provided are pharmaceutical compositions comprising a FAP-activating radiopharmaceutical prodrug and an immune response stimulator. In some embodiments, the immune response stimulator comprises one, two, three, or more immune response stimulators.

[0236] In some embodiments, in addition to administering a FAP-activated radiopharmaceutical prodrug described herein, the method or treatment further comprises administering at least one additional therapeutic agent. The additional therapeutic agent can be administered before, simultaneously with, and / or after administration of the FAP-activated radiopharmaceutical prodrug. Pharmaceutical compositions comprising a FAP-activated radiopharmaceutical prodrug and an additional therapeutic agent are also provided. In some embodiments, the at least one additional therapeutic agent comprises one, two, three, or more additional therapeutic agents.

[0237] Combination therapy with two or more therapeutic agents often uses agents that act by different mechanisms of action, although this is not required. Combination therapy using agents with different mechanisms of action may result in additive or synergistic effects. Combination therapy may allow for lower doses of each agent than those used in monotherapy, thereby reducing toxic side effects and / or increasing the therapeutic index of FAP-activated radiopharmaceutical prodrugs. Combination therapy may reduce the likelihood of resistant cancer cells developing. In some embodiments, combination therapy includes a therapeutic agent that affects the immune response (e.g., enhances or activates the response) and a therapeutic agent that affects tumor / cancer cells (e.g., inhibits or kills them).

[0238] In some embodiments of the methods described herein, the combination of a FAP-activated radiopharmaceutical prodrug described herein with at least one additional therapeutic agent results in additive or synergistic results. In some embodiments, the combination therapy results in an increased therapeutic index of the FAP-activated radiopharmaceutical prodrug. In some embodiments, the combination therapy results in an increased therapeutic index of the additional therapeutic agent. In some embodiments, the combination therapy results in reduced toxicity and / or side effects of the FAP-activated radiopharmaceutical prodrug. In some embodiments, the combination therapy results in reduced toxicity and / or side effects of the additional therapeutic agent.

[0239] Useful classes of therapeutic agents include, for example, antitubulin agents, auristatins, DNA minor groove binders, DNA replication inhibitors, alkylating agents (e.g., cisplatin, mononuclear (platinum), dinuclear (platinum), and trinuclear platinum complexes, and platinum complexes such as carboplatin), anthracyclines, antibiotics, antifolates, antimetabolites, chemotherapy sensitizers, duocarmycins, etoposide, fluorinated pyrimidines, ionophores, lexitropsin, nitrosoureas, platinol, purine antimetabolites, puromycin, radiosensitizers, steroids, taxanes, topoisomerase inhibitors, vinca alkaloids, etc. In some embodiments, the second therapeutic agent is an alkylating agent, antimetabolite, mitotic inhibitor, topoisomerase inhibitor, or angiogenesis inhibitor.

[0240] Therapeutic agents that may be administered in combination with the FAP-activated radiopharmaceutical prodrugs described herein include chemotherapeutic agents. Thus, in some embodiments, a method or treatment involves administering a FAP-activated radiopharmaceutical prodrug of the present disclosure in combination with a chemotherapeutic agent or a cocktail of chemotherapeutic agents. Treatment with a FAP-activated radiopharmaceutical prodrug can occur before, simultaneously with, or after the administration of the chemotherapeutic agent. Combination administration can include simultaneous administration, either in a single pharmaceutical formulation or in separate formulations, or sequential administration, in any order, but within a time period that allows all active agents to simultaneously exert their biological activity. Preparation and administration schedules for such chemotherapeutic agents can be used according to manufacturer's instructions or as empirically determined by one of skill in the art. Preparation and administration schedules for such chemotherapy are also described in The Chemotherapy Source Book, 4th Edition, 2008, MC Perry, Editor, Lippincott, Williams & Wilkins, Philadelphia, PA.

[0241] Chemotherapeutic agents useful in the present disclosure include alkylating agents, such as thiotepa and cyclophosphamide (CYTOXAN); alkylsulfonates, such as busulfan, improsulfan, and piposulfan; aziridines, such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylmelamines, including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolmelamine. ); nitrogen mustards, for example, chlorambucil, chlornaphazine, colofosfamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, nobuenbikinin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosoureas, for example, carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics, for example, aclacinomycin, actinomycin, ausramycin, azaserine, bleomycin, ka Cutinomycin, calicheamicin, carabicin, carminomycin, carzinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogalamycin, olivomycin, peplomycin, potfilomycin, puromycin, queramycin, rodorubicin, streptonigrin, streptozocin, tuberculin Lucidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytosine arabinoside, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU;Androgens such as calsterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; Antiadrenal drugs such as aminoglutethimide, mitotane, trilostane; Folic acid supplements such as folinic acid; Aceglatone; Aldophosphamide glycosides; Aminolevulinic acid; Amsacrine; Bestravcil; Bisantrene; Edatraxate; Defofamine; Demecolcine; Diaziquone; Elformitin; Elliptinium acetate; Etogluside ;Gallium nitrate;Hydroxyurea;Lentinan;Lonidamine;Mitoguazone;Mitoxantrone;Mopidamol;Nitracrine;Pentostatin;Fenamet;Pirarubicin;Podophyllic acid;2-Ethylhydrazide;Procarbazine;PSK;Razoxane;Sizofiran;Spirogermanium;Tenuazonic acid;Triazicon, 2,2',2"-Trichlorotriethylamine;Urethane;Vindesine;Dacarbazine;Mannomustine;Mitobronitol;Mitolactol;Pipobroman;Gasitosine;Arabinoside (Ara- C); taxoids, such as paclitaxel (TAXOL) and doxetaxel (TAXOTERE); chlorambucil; gemcitabine, 6-thioguanine; mercaptopurine; platinum analogs, such as cisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; ibandronate; CPT-11; topoisomerase inhibitors R Chemotherapeutic agents include, but are not limited to, FS2000, difluoromethylolnitine (DMFO), retinoic acid, esperamicin, capecitabine (XELODA), and pharmaceutically acceptable salts, acids, or derivatives of any of the above. Chemotherapeutic agents also include antiestrogens, including, for example, tamoxifen, raloxifene, aromatase-inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, and toremifene (FARESTON);Also included are antihormonal agents that act to regulate or inhibit hormone action on tumors, such as antiandrogens, for example, flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids, or derivatives of any of the above. In certain embodiments, the additional therapeutic agent is cisplatin. In certain embodiments, the additional therapeutic agent is carboplatin.

[0242] In some embodiments of the methods described herein, the chemotherapeutic agent is a topoisomerase inhibitor. A topoisomerase inhibitor is a chemotherapeutic agent that inhibits the action of topoisomerase enzymes (e.g., topoisomerase I or II). Topoisomerase inhibitors include, but are not limited to, doxorubicin HCl, daunorubicin citrate, mitoxantrone HCl, actinomycin D, etoposide, topotecan HCl, teniposide (VM-26), and irinotecan, as well as any pharmaceutically acceptable salt, acid, or derivative thereof. In some embodiments, the additional therapeutic agent is irinotecan.

[0243] In some embodiments, the chemotherapeutic agent is an antimetabolite.Antimetabolites are chemicals that resemble metabolites required for normal biochemical reactions, but have a structure that is sufficiently different to interfere with one or more normal functions of cells, such as cell division.Antimetabolites include, but are not limited to, gemcitabine, fluorouracil, capecitabine, methotrexate sodium, ralitrexed, pemetrexed, tegafur, cytosine arabinoside, thioguanine, 5-azacytidine, 6-mercaptopurine, azathioprine, 6-thioguanine, pentostatin, fludarabine phosphate, and cladribine, as well as pharmaceutically acceptable salts, acids, or derivatives thereof.In some embodiments, the additional therapeutic agent is gemcitabine.

[0244] In some embodiments of the methods described herein, the chemotherapeutic agent is an antimitotic agent, including but not limited to, an agent that binds to tubulin. In some embodiments, the agent is a taxane. In some embodiments, the agent is paclitaxel or docetaxel, or a pharmaceutically acceptable salt, acid, or derivative of paclitaxel or docetaxel. In some embodiments, the agent is paclitaxel (TAXOL), docetaxel (TAXOTERE), albumin-bound paclitaxel (nab-paclitaxel; ABRAXANE), DHA-paclitaxel, or PG-paclitaxel. In certain alternative embodiments, the antimitotic agent comprises a vinca alkaloid, such as vincristine, vinblastine, vinorelbine, or vindesine, or a pharmaceutically acceptable salt, acid, or derivative thereof. In some aspects, the antimitotic agent is an inhibitor of kinesin Eg5 or an inhibitor of a mitotic kinase, such as Aurora A or Plk1. In some embodiments, the additional therapeutic agent is paclitaxel. In some embodiments, the additional therapeutic agent is nab-paclitaxel.

[0245] In some embodiments of the methods described herein, the additional therapeutic agent comprises an agent such as a small molecule. For example, the treatment may involve the combined administration of a FAP-activated radiopharmaceutical prodrug of the present disclosure with a small molecule that acts as an inhibitor of tumor-associated antigens, including, but not limited to, EGFR, HER2 (ErbB2), and / or VEGF. In some embodiments, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in combination with a protein kinase inhibitor selected from the group consisting of gefitinib (IRESSA), erlotinib (TARCEVA), sunitinib (SUTENT), lapatanib, vandetanib (ZACTIMA), AEE788, CI-1033, cediranib (RECENTIN), sorafenib (NEXAVAR), and pazopanib (GW786034B). In some embodiments, the additional therapeutic agent comprises an mTOR inhibitor.

[0246] In some embodiments of the methods described herein, the additional therapeutic agent is a small molecule that inhibits a cancer stem cell pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Notch pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Wnt pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the BMP pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Hippo pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the mTOR / AKR pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the RSPO / LGR pathway.

[0247] In some embodiments of the methods described herein, the additional therapeutic agent comprises a biological molecule such as an antibody. For example, treatment may include co-administration of a FAP-activating radiopharmaceutical prodrug of the present disclosure with an antibody against a tumor-associated antigen, including, but not limited to, an antibody that binds EGFR, HER2 / ErbB2, and / or VEGF. In some embodiments, the additional therapeutic agent is an antibody specific for a cancer stem cell marker. In some embodiments, the additional therapeutic agent is an antibody that binds to a component of the Notch pathway. In some embodiments, the additional therapeutic agent is an antibody that binds to a component of the Wnt pathway. In some embodiments, the additional therapeutic agent is an antibody that inhibits a cancer stem cell pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Notch pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Wnt pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the BMP pathway. In some embodiments, the additional therapeutic agent is an antibody that inhibits β-catenin signaling. In some embodiments, the additional therapeutic agent is an antibody that is an angiogenesis inhibitor (e.g., an anti-VEGF or VEGF receptor antibody). In some embodiments, the additional therapeutic agent is bevacizumab (AVASTIN), ramucirumab, trastuzumab (HERCEPTIN), pertuzumab (OMNITARG), panitumumab (VECTIBIX), nimotuzumab, zalutumumab, or cetuximab (ERBITUX).

[0248] In some embodiments of the methods described herein, the additional therapeutic agent is an antibody that modulates the immune response, hi some embodiments, the additional therapeutic agent is an anti-PD-1 antibody, an anti-LAG-3 antibody, an anti-CTLA-4 antibody, an anti-TIM-3 antibody, or an anti-TIGIT antibody.

[0249] Additionally, treatment with the FAP-activated radiopharmaceutical prodrugs described herein can include combination treatment with other biological molecules, such as one or more cytokines (e.g., lymphokines, interleukins, tumor necrosis factors, and / or growth factors), or can involve surgical removal of tumors, removal of cancer cells, or any other therapy deemed necessary by the treating physician. In some embodiments, the additional therapeutic agent is an immune response stimulator.

[0250] In some embodiments of the methods described herein, the FAP-activated radiopharmaceutical prodrug may be combined with a growth factor selected from the group consisting of adrenomedullin (AM), angiopoietin (Ang), BMP, BDNF, EGF, erythropoietin (EPO), FGF, GDNF, G-CSF, GM-CSF, GDF9, HGF, HDGF, IGF, migration stimulating factor, myostatin (GDF-8), NGF, neurotrophins, PDGF, thrombopoietin, TGF-α, TGF-β, TNF-α, VEGF, P1GF, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-12, IL-15, and IL-18.

[0251] In some embodiments of the methods described herein, the additional therapeutic agent is an immune response stimulator, hi some embodiments, the immune response stimulator is selected from the group consisting of granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin-3 (IL-3), interleukin-12 (IL-12), interleukin-1 (IL-1), interleukin-2 (IL-2), B7-1 (CD80), B7-2 (CD86), 4-1BB ligand, anti-CD3 antibody, anti-CTLA-4 antibody, anti-TIGIT antibody, anti-PD-1 antibody, anti-LAG-3 antibody, and anti-TIM-3 antibody.

[0252] In some embodiments of the methods described herein, the immune response stimulator is selected from the group consisting of a modulator of PD-1 activity, a modulator of PD-L2 activity, a modulator of CTLA-4 activity, a modulator of CD28 activity, a modulator of CD80 activity, a modulator of CD86 activity, a modulator of 4-1BB activity, a modulator of OX40 activity, a modulator of KIR activity, a modulator of Tim-3 activity, a modulator of LAG3 activity, a modulator of CD27 activity, a modulator of CD40 activity, a modulator of GITR activity, a modulator of TIGIT activity, a modulator of CD20 activity, a modulator of CD96 activity, a modulator of IDO1 activity, a cytokine, a chemokine, an interferon, an interleukin, a lymphokine, a member of the tumor necrosis factor (TNF) family, and an immunostimulatory oligonucleotide.

[0253] In some embodiments of the methods described herein, the immune response stimulator is selected from the group consisting of a PD-1 antagonist, a PD-L2 antagonist, a CTLA-4 antagonist, a CD80 antagonist, a CD86 antagonist, a KIR antagonist, a Tim-3 antagonist, a LAG3 antagonist, a TIGIT antagonist, a CD20 antagonist, a CD96 antagonist, and / or an IDO1 antagonist.

[0254] In some embodiments of the methods described herein, the PD-1 antagonist is an antibody that specifically binds to PD-1. In some embodiments, the antibody that binds to PD-1 is KEYTRUDA (MK-3475), pidilizumab (CT-011), nivolumab (OPDIVO, BMS-936558, MDX-1106), MEDI0680 (AMP-514), REGN2810, BGB-A317, PDR-001, or STI-A1110. In some embodiments, the antibody that binds to PD-1 is described in WO2014 / 179664, e.g., an antibody identified as APE2058, APE1922, APE1923, APE1924, APE1950, or APE1963, or an antibody comprising the CDR regions of any of these antibodies. In another embodiment, the PD-1 antagonist is a fusion protein that includes PD-L2, e.g., AMP-224. In another embodiment, the PD-1 antagonist is a peptide inhibitor, e.g., AUNP-12.

[0255] In some embodiments, the CTLA-4 antagonist is an antibody that specifically binds to CTLA-4. In some embodiments, the antibody that binds to CTLA-4 is ipilimumab (YERVOY) or tremelimumab (CP-675,206). In some embodiments, the CTLA-4 antagonist is a CTLA-4 fusion protein, such as KAHR-102.

[0256] In some embodiments, the LAG3 antagonist is an antibody that specifically binds to LAG3. In some embodiments, the antibody that binds to LAG3 is IMP701, IMP731, BMS-986016, LAG525, and GSK2831781. In some embodiments, the LAG3 antagonist comprises a soluble LAG3 receptor, such as IMP321.

[0257] In some embodiments, the KIR antagonist is an antibody that specifically binds to KIR. In some embodiments, the antibody that binds to KIR is lirilumab.

[0258] In some embodiments, the immune response stimulator is selected from the group consisting of a CD28 agonist, a 4-1BB agonist, an OX40 agonist, a CD27 agonist, a CD80 agonist, a CD86 agonist, a CD40 agonist, and a GITR agonist. In some embodiments, the OX40 agonist comprises an OX40 ligand or an OX40-binding portion thereof. For example, the OX40 agonist can be MEDI6383. In some embodiments, the OX40 agonist is an antibody that specifically binds to OX40. In some embodiments, the antibody that binds to OX40 is MEDI6469, MEDI0562, or MOXR0916 (RG7888). In some embodiments, the OX40 agonist is a vector (e.g., an expression vector such as an adenovirus or a virus) capable of expressing an OX40 ligand. In some embodiments, the OX40 expression vector is Delta-24-RGDOX or DNX2401.

[0259] In some embodiments, the 4-1BB (CD137) agonist is a binding molecule such as anticalin. In some embodiments, the anticalin is PRS-343. In some embodiments, the 4-1BB agonist is an antibody that specifically binds to 4-1BB. In some embodiments, the antibody that binds to 4-1BB is PF-2566 (PF-05082566) or urelumab (BMS-663513).

[0260] In some embodiments, the CD27 agonist is an antibody that specifically binds to CD27. In some embodiments, the antibody that binds to CD27 is varlilumab (CDX-1127).

[0261] In some embodiments, the GITR agonist comprises a GITR ligand or a GITR binding portion thereof. In some embodiments, the GITR agonist is an antibody that specifically binds to GITR. In some embodiments, the antibody that binds to GITR is TRX518, MK-4166, or INBRX-110.

[0262] In some embodiments, immune response stimulators include, but are not limited to, cytokines such as chemokines, interferons, interleukins, lymphokines, and members of the tumor necrosis factor (TNF) family, hi some embodiments, immune response stimulators include immunostimulatory oligonucleotides, such as CpG dinucleotides.

[0263] In some embodiments, immune response stimulators include, but are not limited to, an anti-PD-1 antibody, an anti-PD-L2 antibody, an anti-CTLA-4 antibody, an anti-CD28 antibody, an anti-CD80 antibody, an anti-CD86 antibody, an anti-4-1BB antibody, an anti-OX40 antibody, an anti-KIR antibody, an anti-Tim-3 antibody, an anti-LAG3 antibody, an anti-CD27 antibody, an anti-CD40 antibody, an anti-GITR antibody, an anti-TIGIT antibody, an anti-CD20 antibody, an anti-CD96 antibody, or an anti-IDO1 antibody.

[0264] In some embodiments, the FAP-activated radiopharmaceutical prodrugs disclosed herein can be used alone or in conjunction with radiation therapy.

[0265] In some embodiments, the FAP-activated radiopharmaceutical prodrugs disclosed herein can be used alone or in conjunction with targeted therapy. Examples of targeted therapy include hormone therapy, signal transduction inhibitors (e.g., EGFR inhibitors such as cetuximab (Erbitux) and erlotinib (Tarceva)); HER2 inhibitors (e.g., trastuzumab (Herceptin) and pertuzumab (Perjeta)); BCR-ABL inhibitors (such as imatinib (Gleevec) and dasatinib (Sprycel)); ALK inhibitors (e.g., crizotinib (Xalkori) and ceritinib (Zykadia)); BRAF inhibitors (e.g., These include: anti-cancer drugs (e.g., vemurafenib (Zelboraf) and dabrafenib (Tafinlar)), gene expression modulators, apoptosis inducers (e.g., bortezomib (Velcade) and carfilzomib (Kyprolis)), angiogenesis inhibitors (e.g., bevacizumab (Avastin) and ramucirumab (Cyramza)), monoclonal antibodies conjugated to toxins (e.g., brentuximab vedotin (Adcetris) and ado-trastuzumab emtansine (Kadcyla)).

[0266] In some embodiments of the present disclosure, the FAP-activating radiopharmaceutical prodrug of the present disclosure is administered in association with a STING agonist, e.g., as part of a pharmaceutical composition. The cyclic dinucleotide (CDN) cyclic di-AMP (produced by Listeria monocytogenes and other bacteria) and its analogs cyclic di-GMP and cyclic GMP-AMP are recognized by host cells as pathogen-associated molecular patterns (PAMPs) that bind to a pathogen recognition receptor (PRR) known as Stimulator of Interferon Genes (STING). STING is an adaptor protein in the cytoplasm of host mammalian cells that activates the TANK-binding kinase (TBK1)-IRF3 and NF-κB signaling axis, resulting in the induction of IFN-β and other gene products that potently activate innate immunity. STING is now recognized as a component of the host cytosolic surveillance pathway that senses infection by intracellular pathogens and responds by inducing the production of IFN-α, leading to the development of an adaptive defensive pathogen-specific immune response consisting of both antigen-specific CD4+ and CD8+ T cells and pathogen-specific antibodies. U.S. Patent Nos. 7,709,458 and 7,592,326; WO2007 / 054279, WO2014 / 093936, WO2014 / 179335, WO2014 / 189805, WO2015 / 185565, WO2016 / 096174, WO2016 / 145102, WO2017 / 027645, WO2017 / 027646, and WO2017 / 075477; and Yan et al., Bioorg. Med. Chem Lett. 18:5631-4, 2008.

[0267] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrugs of the present disclosure are administered in conjunction with an Akt inhibitor. Exemplary AKT inhibitors include GDC0068 (also known as GDC-0068, ipatasertib, and RG7440), MK-2206, perifosine (also known as KRX-0401), GSK690693, AT7867, triciribine, CCT128930, A-674563, PHT-427, Akti-1 / 2, afuresertib (G SK2110183), AT13148, GSK2141795, BAY1125976, uprosertib (also known as GSK2141795), AKT inhibitor VIII (1,3-dihydro-1-[1-[[4-(6-phenyl-1H-imidazo[4,5-g]quinoxalin-7-yl)phenyl]m-ethyl]-4-piperidinyl]-2H-benzimidazol-2-one), Akt inhibitor X (2-chloro-N,N-diethyl- 10H-phenoxazine-10-butanamine, monohydrochloride), MK-2206 (8-(4-(1-aminocyclobutyl)phenyl)-9-phenyl-[1,2,4]triazolo[3,4-f][-1,6]naphthyridin-3(2H)-one), uprosertib (N-((S)-1-amino-3-(3,4-difluorophenyl)propan-2-yl)-5-chloro-4-(4-chloro-1-methyl-1H-pyrazol-5-yl)furan-2-carboxamide), Patasertib ((S)-2-(4-chlorophenyl)-1-(4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperazin-1-yl)-3-(isopropylamino)propan-1-one), AZD5363 (4-piperidinecarboxamide, 4-amino-N-[(1S)-1-(4-chlorophenyl)-3-hydroxypropyl]-1-(7H-pyrrolo[2,[3-d]p-irimidin-4-yl), perifosine, GSK690693, GDC-0068, tricirbine, CCT128930, A-674563, PF-04691502, AT7867, miltefosine, PHT-427, honokiol, triciribine phosphate, and KP372-1A (10H-indeno[2,1-e]tetrazolo[1,5-b][1,2,4]triazin-10-one), Akt inhibitor IX (CAS 98510-80-6). Additional Akt inhibitors include ATP-competitive inhibitors, such as isoquinoline-5-sulfonamides (e.g., H-8, H-89, NL-71-101), azepane derivatives (e.g., (-)-balanol derivatives), aminofurazans (e.g., GSK690693), heterocycles (e.g., 7-azaindole, 6-phenylpurine derivatives, pyrrolo[2,3-d]pyrimidine derivatives, CCT128930, 3-aminopyrrolidine, anilinotriazole derivatives, spiroindoline derivatives, AZD5363, A-674563 , A-443654), phenylpyrazole derivatives (e.g., AT7867, AT13148), thiophenecarboxamide derivatives (e.g., afuresertib (GSK2110183), 2-pyridimyl-5-amidothiophene derivatives (DC120), uprosertib (GSK2141795); allosteric inhibitors, for example, 2,3-diphenylquinoxaline analogues (e.g., 2,3-diphenylquinoxaline derivatives, triazolo[3,4-f][1,6] naphthyridin-3(2H)-one derivatives (MK-2206), alkylphospholipids (e.g., edelfosine (1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine, ET-18-OCH3), ilmofosine (BM41.440), miltefosine (hexadecylphosphocholine, HePC), perifosine (D-21266), erucylphosphocholine (ErPC), erufosine (ErPC3, erucylphosphohomocholine), indole-3-cal Binol analogues (e.g., indole-3-carbinol, 3-chloroacetylindole, diindolylmethane, diethyl 6-methoxy-5,7-dihydroindolo[2,3-b]carbazole-2,10-dicarboxylate (SR13668), OSU-A9), sulfonamide derivatives (e.g., PH-316, PHT-427), thiourea derivatives (e.g., PIT-1, PIT-2, DM-PIT-1, N-[(1-methyl-1H-pyrazol-4-yl)carbonyl]-N'-(3-bromophenyl)-thiourea), purine derivatives (e.g., triciribine (TCN, NSC)), 154020), triciribine monophosphate active analogue (TCN-P), 4-amino-pyrido[2,3-d]pyrimidine derivative API-1, 3-phenyl-3H-imidazo[4,5-b]pyridine derivative, ARQ 092), BAY 1125976, 3-methyl-xanthine, quinoline-4-carboxamide, 2-[4-(cyclohexa-1,3-dien-1-yl)-1H-pyrazol-3-yl]phenol, 3-oxo-tirucaric acid, 3α- and 3β-acetoxy-tirucaric acid, acetoxy-tirucaric acid; and irreversible inhibitors, such as natural products, antibiotics, lactoquinomycin, frenolicin B, calafungin, medelmicin, Boc-Phe-vinyl ketone, 4-hydroxynonenal (4-HNE), 1,6-naphthyridinone derivatives, and imidazo-1,2-pyridine derivatives.

[0268] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in conjunction with a MEK inhibitor. Exemplary MEK inhibitors include AZD6244 (Selumetinib), PD0325901, GSK1120212 (Trametinib), U0126-EtOH, PD184352, RDEA119 (Rafametinib), PD98059, BIX02189, MEK162 (Binimetinib), AS-703026 (Pimasertib), SL-327, BIX02188, AZD8330, TAK-733, Cobimetinib, and PD318088.

[0269] In some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with both an anthracycline, such as doxorubicin (including pegylated liposomal doxorubicin), and cyclophosphamide.

[0270] In some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with both an anti-CD20 antibody and an anti-CD3 antibody, or a bispecific CD20 / CD3 binding agent (including a CD20 / CD3 BiTE).

[0271] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in conjunction with a CD73 inhibitor, a CD39 inhibitor, or both. These inhibitors can be CD73 or CD39 binding agents (such as antibodies, antibody fragments, or antibody mimetics) that inhibit ectonucleosidase activity. The inhibitor can be a small molecule inhibitor of ectonucleosidase activity, such as 6-N,N-diethyl-β-γ-dibromomethylene-D-adenosine-5′-triphosphate trisodium salt hydrate, PSB069, or PSB06126.

[0272] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrugs of the present disclosure are administered in association with an inhibitor of poly ADP-ribose polymerase (PARP). Exemplary PARP inhibitors include Olaparib, Niraparib, Rucaparib, Talazoparib, Veliparib, CEP9722, MK4827, and BGB-290.

[0273] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with an oncolytic virus. An exemplary oncolytic virus is Talimogene Laherparepvec.

[0274] In some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with a CSF-1 antagonist, such as an agent that binds to CSF-1 or CSF1R and inhibits the interaction of CSF-1 with CSF1R on macrophages. Exemplary CSF-1 antagonists include Emactuzumab and FPA008.

[0275] In some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with an anti-CD38 antibody. Exemplary anti-CD38 antibodies include daratumumab and isatuximab.

[0276] In some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in association with an anti-CD40 antibody. Exemplary anti-CD40 antibodies include Selicrelumab and Dacetuzumab.

[0277] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in conjunction with an inhibitor of anaplastic lymphoma kinase (ALK). Exemplary ALK inhibitors include alectinib, crizotinib, and ceritinib.

[0278] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in conjunction with a multikinase inhibitor that inhibits one or more selected from the group consisting of VEGFR, PDGFR, and FGFR family members, or an anti-angiogenic inhibitor. Exemplary inhibitors include axitinib, cediranib, linifanib, motesanib, nintedanib, pazopanib, ponatinib, regorafenib, sorafenib, sunitinib, tivozanib, vatalanib, LY2874455, or SU5402.

[0279] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrugs of the present disclosure are administered in conjunction with one or more vaccines intended to stimulate an immune response to one or more predetermined antigens. The antigens may be administered directly to the individual or may be expressed in the individual from, for example, tumor cell vaccines (e.g., GVAX), dendritic cell vaccines, DNA vaccines, RNA vaccines, virus-based vaccines, bacterial or yeast vaccines (e.g., Listeria monocytogenes or Saccharomyces cerevisiae), which may be autologous or allogeneic. See, for example, Guo et al., Adv. Cancer Res. 2013; 119: 421-475; Obeid et al., Semin Oncol. 2015 August; 42(4): 549-561. The target antigen may also be a fragment or fusion polypeptide containing an immunologically active portion of the antigens listed in the table.

[0280] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrugs of the present disclosure are administered in association with one or more antiemetic drugs, including, but not limited to, casopitant (GlaxoSmithKline), netupitant (MGI-Helsinn) and other NK-1 receptor antagonists, palonosetron (sold as Aloxi by MGI Pharma), aprepitant (sold as Emend by Merck and Co.; Rahway, NJ), diphenhydramine (sold as Benadryl by Pfizer; New York, NY), hydroxyzine (sold as Atarax by Pfizer; New York, NY), metoclopramide (sold as Reglan by A.H. Robins Co.; Richmond, Va.), lorazepam (sold as Ativan by Wyeth; Madison, NJ), alprazolam (sold as Xanax by Pfizer; New York, NY), New York, NY), haloperidol (sold as Haldol by Ortho-McNeil; Raritan, NJ), droperidol (Inapsine), dronabinol (sold as Marinol by Solvay Pharmaceuticals, Inc.; Marietta, Ga.), dexamethasone (sold as Decadron by Merck and Co.; Rahway, NJ), methylprednisolone (sold as Medrol by Pfizer; New York, NY), prochlorperazine (sold as Compazine by Glaxosmithkline; Research Triangle Park, N.C.), granisetron (sold as Kytril by Hoffmann-La Roche Inc.; Nutley, NJ), ondansetron (sold as Zofran by Glaxosmithkline; Research Triangle Park, N.C.), dolasetron (sold as Anzemet by Sanofi-Aventis; New York, NY). York, NY), tropisetron (sold as Navoban by Novartis; East Hanover, NJ).

[0281] Other side effects of cancer treatment include red and white blood cell deficiencies. Thus, in some embodiments of the present disclosure, a FAP-activated radiopharmaceutical prodrug is administered in association with an agent that treats or prevents such deficiencies, such as filgrastim, PEG-filgrastim, erythropoietin, epoetin alfa, or darbepoetin alfa.

[0282] In some embodiments of the present disclosure, the FAP-activated radiopharmaceutical prodrug of the present disclosure is administered in conjunction with anti-cancer radiation therapy. For example, in some embodiments of the present disclosure, the radiation therapy is external beam radiation therapy (EBT), a method for delivering a beam of high-energy X-rays to the location of a tumor. The beam is generated outside the patient's body (e.g., by a linear accelerator) and targeted to the tumor site. These X-rays can destroy cancer cells, and careful treatment planning can spare surrounding normal tissue. No radioactive source is placed inside the patient's body. In some embodiments of the present disclosure, the radiation therapy is proton beam therapy, a type of conformal radiation therapy that bombards diseased tissue with protons instead of X-rays. In some embodiments of the present disclosure, the radiation therapy is conformal external beam radiation therapy, a procedure that uses advanced technology to tailor radiation therapy to an individual's anatomy. In some embodiments of the present disclosure, the radiation therapy is brachytherapy, a temporary placement of radioactive materials inside the body, usually used to give an area an extra dose or boost of radiation. [Example]

[0283] V. Example Example 1: [ka] The synthetic scheme for the preparation of compound 7885 is shown in Figure 1; in Figure 1, i. BrCH2Cl, NaHCO3; ii. TEA, NaI; iii. TFA-DCM; iv. DOTA-PNP.

[0284] Example 2: [ka] The synthetic scheme for the preparation of compound 6885 is shown in Figure 2; in Figure 2, i. triphosgene, Py; ii. Lys(Fmoc)-OtBu, DIEA, flash column purified; iii. 50% TEA in DCM; iv. Fmoc-L-2-Nal-OH, HATU, DIEA, DMF, flash column purified; v. 50% TEA in DCM; vi. N-Boc-tranexamic acid, HBTU, DIEA, DMF, flash column purified; vii. Pd(PPh3)4, flash column purified; viii. BrClCHCH3, Cs2CO3; ix. HATU, DIEA; x. Pd(PPh3)4; xi. Cs2CO3; xii. TFA-DCM; xiii. DOTA-PNP.

[0285] Example 3: [ka] The synthesis scheme for compound 6879 is shown in Figure 3. In Figure 3, i. Pd[PPh3]4, morpholine, DCM; ii. Fmoc-L-2-Nal-OH, HBTU, DIEA, DMF; iii. 50% piperidine in DMF; iv. N-Fmoc-tranexamic acid, HBTU, DIEA, DMF; v. 50% piperidine in DMF; vi. TFA, TIPS; vii. DOTA-PNP.

[0286] Example 4: [ka] The synthesis scheme of compound 6880 is shown in Figure 4. In Figure 4, i. Pd[PPh3]4, morpholine, DCM; ii. Fmoc-L-2-Nal-OH, HBTU, DIEA, DMF; iii. 50% piperidine in DMF; iv. N-Fmoc-tranexamic acid, HBTU, DIEA, DMF; v. 50% piperidine in DMF; vi. TFA, TIPS; vii. DOTA-PNP.

[0287] Example 5: [ka] The synthetic scheme of compound 6886 is shown in Figure 5; in Figure 5, i. triphosgene, Py; ii. Lys(Fmoc)-OtBu, DIEA; iii. 50% piperidine in DMF; iv. Fmoc-L-2-Nal-OH, HBTU, DIEA, DMF; v. 50% piperidine in DMF; vi. N-Boc-tranexamic acid, HBTU, DIEA, DMF; vii. Pd(PPh3)4; viii. BrClCH2, Cs2CO3; ix. HATU, DIEA; x. Pd(PPh3)4; xi. Cs2CO3; xii. TFA-DCM; xiii. DOTA-PNP.

[0288] Example 6: Additional Compounds and Synthesis [ka]

[0289] [ka]

[0290] [ka]

[0291] [ka]

[0292] [ka]

[0293] [ka]

[0294] [ka]

[0295]

change

[0296]

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[0297]

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[0298]

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[0299]

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[0300]

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[0301]

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[0302]

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[0303]

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[0304]

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[0305] [ka]

[0306] [ka]

[0307] Example 7: IC50 assay of 7028P and FAP-activated 7028A, 7028B and 7028C Objective: The purpose of this study was to demonstrate the affinity of 7028P and 7028A / B / C for binding to prostate-specific antigen (PSMA) by measuring inhibition of PSMA enzymatic activity. Enzyme activity was measured using acetyl-Asp-Glu as a substrate, monitoring the generation of primary amino groups generated by enzymatic cleavage of the peptide bond. The amino groups were detected using the fluoroaldehyde o-phthalaldehyde, which, together with mercaptoethanol, forms a fluorescent adduct with primary amines.

[0308] The structures of 7028P and 7028A / B / C are shown below. [ka]

[0309] method material: ·rhPSMA(R&D systems, 4334-ZN-010) Assay buffer: 50mM HEPES pH 7.5, 100mM NaCl Ac-Asp-Glu (Sigma, A5930) 2-PMPA (2-(phosphonomethyl)-pentanedioic acid, Sigma, SML1612) Fluoroaldehyde OPA (Thermo Fisher Scientific, 26025) ·7028P, 7028A, 7028B&7028C(Tufts) Corning 96-well flat-bottom polystyrene NBS (Fisher, 07-201-203)

[0310] Device: Molecular Devices M2e Plate Reader

[0311] procedure: 1. rhPSMA was dissolved in assay buffer at 0.4 μg / mL. 2. Ac-Asp-Glu was dissolved in assay buffer at 80 μM. 3. Stock solutions of inhibitors (7028P, 7028A, 7028B, and 7028C) were prepared at 100 μM in DMSO. 4. Inhibitor stocks were diluted by adding 160 μL to 240 μL of assay buffer for a 40 μM working stock. A series of 10× dilutions were made to prepare 4× concentrated samples of each inhibitor as shown below. [Table 1] 5. Inhibitor and substrate were mixed by combining 100 μL of inhibitor with 100 μL of 80 μM substrate. 6. The reaction was initiated by adding an equal volume (200 μL) of 0.4 μg / mL rhPMSA. The reaction mixture was incubated at 37°C for 60 minutes. a. Enzyme concentration in the reaction: 0.2 μg / mL b. Substrate concentration in the reaction: 20 μM 7. A blank sample containing only assay buffer and substrate, and an inhibitor-free control reaction sample (100 μL buffer, and 100 μL of 80 μM substrate and 200 μL of 0.4 mg / μL rhPSMA) were prepared and reacted in the same manner as the inhibitor samples. 8. The reaction was stopped by heating in a boiling water bath for 5 minutes. 9. 100 μL of each sample was placed in triplicate wells of a 96-well plate. 10. 100 μL of Fluoroaldehyde OPA Reagent was added to each well and mixed. 11. Fluorescence was measured at excitation 330 nm and emission 450 nm. 12. Data were normalized to the maximum value and IC50 was determined using the Prism 9 equation "log(inhibitor) vs. response (3 parameters)".

[0312] The results are shown in Figure 6.

[0313] Example 8: Additional Syntheses and Compounds (FAP-Activated Folic Acid / Methotrexate [MTX] Prodrug Conjugates) [ka] [ka] The synthetic scheme for 6970B and 6970B-ester isomers is shown in FIG.

[0314] [ka] The synthesis scheme of 7014 is shown in Figure 8.

[0315] [ka] [ka] The synthetic scheme of 7366P5 is shown in FIG. 9, and the synthetic scheme of 7366 is shown in FIG.

[0316] Example 9: FAP activation of 6970B isomers 1 & 2 and 7366 The results of FAP activation of 6970B isomers 1 & 2 and 7366 with 100 μM substrate and 50 nM FAP are shown in FIG.

[0317] The LC / MS spectrum of 6970B Isomer 1 at 0.1 mM in FAP assay buffer is shown in FIG. instrumentation: ·Agilent 1290 HPLC / 6460 Triple Quad LC / MS Column: Zorbax Eclipse Plus C18, 4.6 x 50 mm, 1.8 μm ·HPLC method: ·Mobile phase A: 0.1% TFA aqueous solution ·Mobile phase B: 0.08% TFA in ACN ·Flow rate: 0.5mL / min Gradient: 0-3 min, 10% B; 25 min, 98% B.

[0318] The LC / MS spectrum of 6970B Isomer 2 (0.1 mM in FAP assay buffer) is shown in FIG. instrumentation: ·Agilent 1290 HPLC / 6460 Triple Quad LC / MS Column: Zorbax Eclipse Plus C18, 4.6 x 50 mm, 1.8 μm ·HPLC method: ·Mobile phase A: 0.1% TFA aqueous solution ·Mobile phase B: 0.08% TFA in ACN ·Flow rate: 0.5mL / min Gradient: 0-3 min, 10% B; 25 min, 98% B.

[0319] The LC / MS spectrum of the 6970B mixture containing isomers 1 & 2 is shown in FIG. 6970B isomer 1 and isomer 2 are co-injected. instrumentation: ·Agilent 1290 HPLC / 6460 Triple Quad LC / MS Column: Zorbax Eclipse Plus C18, 4.6 x 50 mm, 1.8 μm ·HPLC method: ·Mobile phase A: 0.1% TFA aqueous solution ·Mobile phase B: 0.08% TFA in ACN ·Flow rate: 0.5mL / min Gradient: 0-3 min, 10% B; 25 min, 98% B.

[0320] The LC / MS spectrum of 7366 is shown in FIG. 0.1 mM 7366 in FAP buffer, T = 0 min Peak assignments: Peak @ 12.6 min, MV. 578.4 / 115.4, 7366, Ref. LC / MSS (Peak 1) instrumentation: ·Agilent 1290 HPLC / 6460 Triple Quad LC / MS Column: Zorbax Eclipse Plus C18, 4.6 x 50 mm, 1.8 μm ·HPLC method: ·Mobile phase A: 0.1% TFA aqueous solution Mobile phase B: 0.1% TFA in ACN ·Flow rate: 0.5mL / min Gradient: 0-3 min, 10% B; 12 min, 26% B; 12-15 min, 98% B.

Claims

1. A fibroblast activation protein (FAP)-activated theranostic prodrug represented by Formula II, or a pharma- ceutically acceptable salt thereof: 【Chemistry 1】 [In the formula, A represents a 5-8 membered heterocycle; R １０ is an amino terminal blocking group; R １２ is hydrogen, or (C １ ~C ６ ) alkyl; R １３ is hydrogen, or (C １ ~C ６ ) alkyl, which may be straight or branched; R １４ is independently for each occurrence -(C １ ~C ６ ) Alkyl, -OH, -NH ２ or halogen; p is an integer from 0 to 6; L is a bond or a self-eliminating linker which upon cleavage by FAP releases NH 2 -L-R; R is a ligand-targeted theranostic moiety that includes a ligand for binding to a cellular target and a chelator for chelating a radioactive moiety.

2. 2. The FAP-activated theranostic prodrug of claim 1, represented by Formula III: 【Chemistry 2】

3. 3. The FAP-activated theranostic prodrug of claim 2, represented by Formula IV, or a pharma- ceutically acceptable salt thereof: 【Chemistry 3】

4. (i) R １３ is methyl, and / or (ii) R 12 is H; and / or (iii) p is 0. The FAP-activated theranostic prodrug of any one of claims 1 to 3.

5. 3. The FAP-activated theranostic prodrug of claim 2, represented by Formula VI, or a pharma- ceutically acceptable salt thereof. 【Chemistry 4】

6. R 10 is R 11 (C═X)—; Where: X is O or S; R 11 -(C═X) taken together represent an acyl N-terminal blocking group; or R 11 is —(C 1 -C 10 )alkyl, —(C 1 -C 10 )alkoxy, —(C 1 -C 10 )alkyl-C(O)—OH, —(C 1 -C 10 )alkenyl-C(O)—OH, —(C 1 -C 10 )alkyl-C(O)—(C 1 -C 10 )alkyl, —(C 3 -C 8 )cycloalkyl, —(C 3 -C 8 )cycloalkyl(C 1 -C 10 )alkyl, —(C 6 -C 14 )aryl, —(C 6 -C 14 )aryl(C 1 -C 10 )alkyl, —O—(C 1 -C 4 )alkyl-(C 6 -C 14 )aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl(C 1 -C 10 ). ) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; or R 11 is -(AA) n -(C 1 -C 10 )alkyl, -(AA) n -(C 1 -C 10 )alkoxy, -(AA) n -(C 1 -C 10 )alkyl-C(O)-(C 1 -C 10 )alkyl, -(AA) n -(C 3 -C 8 )cycloalkyl, -(AA) n -(C 3 -C 8 )cycloalkyl(C 1 -C 10 )alkyl, -(AA) n -(C 6 -C 14 )aryl, -(AA) n -(C 6 -C 14 )aryl(C 1 -C 10 )alkyl, -(AA) n -5-10 membered heteroaryl, or -(AA) n -5-10 membered heteroaryl(C 1 -C 10) alkyl, where R 11 is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; AA, independently for each occurrence, is an amino acid residue; n is an integer from 1 to 5 The FAP-activated theranostic prodrug of any one of claims 1 to 5.

7. A fibroblast activation protein (FAP)-activated theranostic prodrug represented by formula VII, or a pharma- ceutically acceptable salt thereof: 【Chemistry 5】 [In the formula, R is a ligand-targeted theranostic moiety that includes a ligand for binding to a cellular target and a chelator for chelating a radioactive moiety; R １１ is -(C １ ~C １０ ) alkyl, -(C １ ~C １０ ) alkoxy, -(C １ ~C １０ ) alkyl-C(O)-OH, -(C １ ~C １０ ) alkenyl-C(O)-OH, -(C １ ~C １０ ) alkyl-C(O)-(C １ ~C １０ ) alkyl, -(C ３ ~C ８ ) cycloalkyl, -(C ３ ~C ８ ) Cycloalkyl(C １ ~C １０ ) alkyl, -(C ６ ~C １４ ) aryl, -(C ６ ~C １４ ) Aryl (C １ ~C １０ ) alkyl, —O—(C １ ~C ４ ) alkyl-(C ６ ~C １４ ) aryl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C １ ~C １０ ) alkyl, where R １１ is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, alkoxy, carboxy, cyano, amino, nitro, and thio; L is a bond or is cleaved by FAP to give NH ２ -LR is released after which it is a self-eliminating linker.

8. 7. The FAP-activated theranostic prodrug of claim 6, wherein X is O.

9. R １１ is -(C １ ~C １０ ) alkyl, -(C １ ~C １０ ) alkoxy, -(C ３ ~C ８ ) cycloalkyl, -(C ６ ~C １４ ) aryl, -(C ６ ~C １４ ) Aryl (C １ ~C １０ 9. The FAP-activated theranostic prodrug of any one of claims 6 to 8, wherein R is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27

10. R １１ teeth, 【Chemistry 6】 10. The FAP-activated theranostic prodrug of claim 9,

11. The FAP-activated theranostic prodrug of any one of claims 6 to 10, wherein n is equal to 1 and AA is a serine residue.

12. The FAP-activated theranostic prodrug of any one of claims 6 to 8, wherein n is 1 or 2.

13. R １１ (C １ ~C １０ ) alkyl, (C １ ~C １０ ) alkoxy, (C １ ~C １０ ) alkyl-C(O)-(C １ ~C １０ ) alkyl, (C ３ ~C ８ ) cycloalkyl, (C ３ ~C ８ ) Cycloalkyl(C １ ~C １０ ) alkyl, (C ６ ~C １４ ) aryl, (C ６ ~C １４ ) Aryl (C １ ~C １０ ) alkyl, 5- to 10-membered heteroaryl, or 5- to 10-membered heteroaryl (C １ ~C １０ ) alkyl, where R １１ is optionally substituted with one or more substituents independently selected from the group consisting of halo, hydroxy, carboxy, cyano, amino, nitro, and thio; R １２ is hydrogen; R １３ (C １ ~C ６ ) alkyl; R １４ is absent, or p is 2 and R １４ is a halogen for each occurrence; L is a bond or -N(H)-L- is a self-elimination linker The FAP-activated theranostic prodrug of any one of claims 6 to 8.

14. C(O)-R １１ The FAP-activated theranostic prodrug of claim 8, wherein:

15. C(O)-R １１ is a formyl, acetyl, propionyl, butryl, oxalyl, malonyl, succinyl, glutaryl, adipoyl, acryloyl, maleoyl, fumaroyl, glycoloyl, lactoyl, pyruvoyl, glyceroyl, maloyl, oxaloacetyl, benzoyl, trifluoroacetyl or methoxysuccinyl group.

16. R １１ Ha-(CH ２ ) ｍ -R １１ａ and R １１ａ The FAP-activated theranostic prodrug of any one of claims 6 to 8, wherein is a 5-10 membered aryl or heteroaryl group and m is an integer from 1 to 6.

17. said aryl is selected from the group consisting of benzyl, naphthalenyl, phenanthrenyl, phenolyl, and anilinyl; 17. The FAP-activated theranostic prodrug of claim 16, wherein said heteroaryl is selected from the group consisting of pyrryl, furyl, thiophenyl (a / k / a thienyl), imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrazolyl, pyridinyl, pyrazinyl, pyridazinyl, and pyrimidinyl.

18. (i) L is a self-elimination linker, the self-elimination linker being 【Chemistry 7】 【Chemistry 8】 is selected from the group consisting of During the ceremony, R ａ is hydrogen, alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl; R ｂ is halogen, alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, or heteroaryl; h is an integer from 0 to 8, as valences permit; i is an integer from 1 to 6; or (ii) L is 【Chemistry 9】 and wherein R 1 is hydrogen, unsubstituted or substituted C 1-3 alkyl, or unsubstituted or substituted heterocyclyl; (iii) L is 【Chemistry 10】 or (iv) L is 【Chemistry 11】 is selected from During the ceremony, U is O, S or NR 6 ; Q is CR 4 or N; V 1 , V 2 and V 3 are independently CR 4 or N, with the proviso that for Formulas II and III, at least one of Q, V 1 and V 2 is N; T is NH, NR 6 , O, or S attached to said therapeutic moiety; R 1 , R 2 , R 3 and R 4 are independently H, F, Cl, Br, I, OH, -N(R 5 ) 2 , -N(R 5 ) 3 + , C 1 to C 8 alkyl halide, carboxylate, sulfate, sulfamate, sulfonate, -SO 2 R 5 , -S(═O)R 5 , -SR 5 , -SO 2 N(R 5 ) 2 , -C(═O)R 5 , -CO 2 R 5 , -C(═O)N(R 5 ) 2 , -CN, -N 3 , -NO 2 , C 1 to C 8 alkoxy, C 1 to C 8 halo-substituted alkyl, polyethyleneoxy, phosphonate, phosphate, C 1 to C 8 alkyl, C 1 to C 8 substituted alkyl, C 2 -C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C20 aryl, C6-C20 substituted aryl, C1-C20 heterocycle, and C1-C20 substituted heterocycle; or R2 and R3 taken together form a carbonyl (=O), or a spirocarbocycle of 3 to 7 carbon atoms; R 5 and R 6 are independently selected from H, C 1 -C 8 alkyl, C 1 -C 8 substituted alkyl, C 2 -C 8 alkenyl, C 2 -C 8 substituted alkenyl, C 2 -C 8 alkynyl, C 2 -C 8 substituted alkynyl, C 6 -C 20 aryl, C 6 -C 20 substituted aryl, C 1 -C 20 heterocycle, and C 1 -C 20 substituted heterocycle; wherein C 1 -C 8 substituted alkyl, C 2 -C 8 substituted alkenyl, C 2 -C 8 substituted alkynyl, C 6 -C 20 substituted aryl, and C 2 -C 20 substituted heterocycle are independently selected from F, Cl, Br, I, OH, —N(R 5 ) 2 , —N(R 5 ) 3 + , C 1 -C 8 alkyl halide, carboxylate, sulfate, sulfamate, sulfonate, C 1 -C 8 alkyl sulfonate, C 1 -C 8 alkyl amino, 4-dialkylaminopyridinium, C 1 -C 8 alkyl hydroxyl, C 1 -C 8 alkyl thiol, -SO 2 R 5 , -S(═O)R 5 , -SR 5 , -SO 2 N(R 5 ) 2 , -C(═O)R 5 , -CO 2 R 5 , -C(═O)N(R 5 ) 2 , -CN, -N 3 , -NO 2 , C 1 -C 8 alkoxy, C 1 -C 8 trifluoroalkyl, C 1 -C 8 alkyl, C 3 -C 12 carbocycle, C 6 -C 20 aryl, C 2 -C 20 Substituted with one or more substituents selected from heterocycles, polyethyleneoxy, phosphonates, and phosphates; or (v) The FAP-activated theranostic prodrug of any one of claims 1 to 17, wherein L is selected from -NH-(CH 2 ) 4 -C(═O)-, or -NH-(CH 2 ) 3 -C(═O)-, p-aminobenzyloxycarbonyl (PABC), 2,4-bis(hydroxymethyl)aniline, or benzyloxycarbonyl.

19. The ligand-targeting theranostic moiety (R) is -TM-L １ -R ２０ is represented by TM represents a ligand targeting moiety that selectively binds to a cell surface feature on a target cell; L １ represents a bond or a linker; R ２０ The FAP-activated theranostic prodrug of any one of claims 1 to 18, wherein represents a chelating agent.

20. (i) the ligand targeting moiety is a folate receptor ligand; or (ii) the ligand targeting moiety is somatostatin or a somatostatin analog; or (iii) The FAP-activated theranostic prodrug of claim 19, wherein the ligand targeting moiety is an αIIbβ3 targeting ligand.

21. (i) the ligand targeting moiety is a folic acid or folic acid analog selected from etharfolatide, vintafolide, leucovorin, and methotrexate; or (ii) the ligand targeting moiety is a somatostatin or somatostatin analogue selected from octreotate, octreotide, pentetreotide, lanreotide, vapreotide, pasireotide, seglitide, venereotide, KE-108, SDZ-222-100, Sst3-ODN-8, CYN-154806, JR11, J2156, SRA-880, ACQ090, P829, SSTp-58, SSTp-86, BASS, or somatoprim; or (iii) The FAP-activated theranostic prodrug of claim 20, wherein the ligand targeting moiety is an RGD or RGD analog selected from cyclo(-Arg-Gly-Asp-D-PheVal-)["c(RGDfV)"], c(RGDfK), c(RGDfC), c(RADfC), c(RADfK), c(RGDfE), c(RADfE), RGDSK, RADSK, RGDS, c(RGDyC), c(RADyC), c(RGDyE), c(RGDyK), c(RADyK), H-E[c(RGDyK)]2, EMD12194, DMP728, DMP757, and SK&F107260.

22. The ligand-targeting theranostic moiety (R) is 【Chemistry 12】 and In the formula, R ３０ 20. The FAP-activated theranostic prodrug of claim 19, wherein, independently for each occurrence, represents hydrogen or lower alkyl.

23. -L １ -R ２０ teeth, 【Chemistry 13】 is represented by In the formula, R ３１ is -(CH ２ ) ｐ -aryl or -(CH ２ ) ｐ -heteroaryl, and p is 0, 1, 2, 3 or 4.

24. R ３１ Ha-CH ２ -aryl, where the aryl group is C ６ ~C １２ 24. The FAP-activated theranostic prodrug of claim 23, which is aryl and is a monocyclic ring or a bicyclic fused ring.

25. -L １ -R ２０ teeth, 【Chemistry 14】 The FAP-activated theranostic prodrug of any one of claims 19 to 22, represented by:

26. The ligand-targeting theranostic moiety (R) comprises a folic acid or folic acid analog selected from: 【Chemistry 15】 During the ceremony, R ２１ represents H, R ２２ is -NH-(CH ２ ) ｑ -R ２０ , -NH-(CH ２ ) ｑ -NH-C(O)-(CH ２ ) ｑ -R ２０ or -NH-(CH ２ ) ｑ -C(O)-(CH ２ ) ｑ -R ２０ represents; or R ２２ represents H, R ２１ is -NH-(CH ２ ) ｑ -R ２０ or -NH-(CH ２ ) ｑ -C(O)-(CH ２ ) ｑ -R ２０ represents; or R ２１ Or R ２２ One of them represents H and the other represents 【Chemistry 16】 is selected from the group R ２３ is H, -CH ３ , -CH ２ CH ３ or -CO ２ Represents H; 20. The FAP-activated theranostic prodrug of claim 19, wherein q is, independently for each occurrence, 0, 1, 2, 3 or 4.

27. ​​(i) R ２１ is -NH-CH ２ -R ２０ , —NH—CH ２ -C(O)-R ２０ , -NH-C(O)-CH ２ -R ２０ , —NH—CH ２ —C(O)—CH ２ -R ２０ or -NH-(CH ２ ) ２ -NH-C(O)-CH ２ -R ２０ represents R ２２ represents H; or (ii) A FAP-activated theranostic prodrug according to claim 26, wherein R 21 represents H and R 22 represents -NH-CH 2 -R 20 , -NH-CH 2 -C(O)-R 20 , -NH-C(O)-CH 2 -R 20 , -NH-CH 2 -C(O)-CH 2 -R 20 , or -NH-(CH 2 ) 2 -NH-C(O)-CH 2 -R 20 .

28. The ligand-targeting theranostic moiety (R) is 【Chemistry 17】 20. The FAP-activated theranostic prodrug of claim 19, wherein

29. R is 【Chemistry 18】 【Chemistry 19】 20. The FAP-activated theranostic prodrug of claim 19 selected from:

30. R is 【Chemistry 20】 20. The FAP-activated theranostic prodrug of claim 19 selected from:

31. 【Chemical 21】 【Chemical 22】 【Chemistry 23】 13. The FAP-activated theranostic prodrug of claim 1, having a structure selected from:

32. 【Catalogue 24】 13. The FAP-activated theranostic prodrug of claim 1, having a structure selected from:

33. The prodrug comprises a radioisotope chelated thereto, the radioisotope being １８ F. ４３ K. ４７ Sc, ５１ Cr, ５７ Co, ５８ Co, ５９ Fe, ６４ Cu, ６７ Cu, ６７ G.A. ６８ G.A. ７１ Ge, ７２ As, ７２ Se, ７５ B, ７６ B, ７７ As, ７７ B, ８１ Rb, ８８ Y. ９０ Y. ９７ Ru, ９９ｍ Tc, １００ Pd, １０１ｍ Rh, １０３ Pb, １０５ Rh, １０９ Pd, １１１ Ag, １１１ In, １１３ In, １１９ Sb １２１ Sn, １２３ I, １２４ I, １２５ I, １２７ Cs, １２８ Ba, １２９ Cs, １３１ Cs, １３１ I, １３９ La, １４０ La, １４２ P.R. １４３ P.R. １４９ P.M., １５１ EU, １５３ EU, １５３ Sm, １５９ G, １６１ Tb, １６５ Dy, １６６ Ho, １６９ EU, １７５ Yb, １７７ Lu, １８６ Re, １８８ Re, １８９ Re, １９１ Os, １９３ Pt, １９４ Ir, １９７ Hg, １９８ Au, １９９ Ag, １９９ Au, ２０１ Tl, ２０３ Pb, ２１１ At, ２１２ Bi, ２１２ Pb, ２１３ Bi, ２２５ Ac and ２２７ Th 33. The FAP-activated theranostic prodrug of any one of claims 1 to 32, selected from the group consisting of:

34. k for cleavage by FAP is at least 10-fold greater than that by prolyl endopeptidase ｃａｔ / K ｍ 33. The FAP-activated theranostic prodrug of any one of claims 1 to 32, having the formula:

35. 35. A pharmaceutical composition comprising the FAP-activated theranostic prodrug of any one of claims 1 to 34, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

36. 35. A composition for treating a disorder characterized by upregulation of fibroblast activation protein (FAP), comprising a FAP-activated theranostic prodrug of any one of claims 1 to 34, or a pharma- ceutically acceptable salt thereof.

37. 37. The composition of claim 36, wherein the disorder characterized by upregulation of FAP is cancer.

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