Heterobivalent and homobivalent agents targeting fibroblast activation protein alpha and / or prostate-specific membrane antigen

Abstract

Disclosed are diagnostic imaging and radiotherapeutic agents that target fibroblast activation protein-alpha (FAP-α) and / or prostate-specific membrane antigen (PSMA) and their use in the diagnostic imaging and treatment of diseases and disorders associated with FAP-α and / or PSMA.

Description

Statement of Government Interest

[0001] This invention was made with government support under Grants CA134675 and EB024495 awarded by the National Institutes of Health. The Government has certain rights in this invention. [Background technology]

[0002] Prostate-specific membrane antigen (PSMA) acts as a target for a wide variety of cancers, including prostate cancer and clear cell renal cell carcinoma. In addition to expression in tumor epithelium, PSMA is also expressed in the neovasculature of essentially all solid tumors. On the other hand, fibroblast activation protein alpha (FAP-α) is expressed in cancer-associated fibroblasts, a key driver of the malignant phenotype, and is expressed in nearly all cancers as well. Thus, PSMA and FAP-α can serve as markers for different aspects of cancer, namely epithelium and tumor microenvironment. A platform that simultaneously targets both PSMA and FAP-α would not only be able to image, but if properly functionalized with therapeutic agents, it would also be able to treat PSMA- and FAP-α-associated cancers in a manner superior to one agent alone. Summary of the Invention

[0003] In certain embodiments, the subject matter of the present disclosure has formula (I): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is a targeting moiety for prostate-specific membrane antigen (PSMA) or FAP-α, and when A and B are each a targeting moiety for FAP-α, they can be the same or different; C is any optically or radiolabeled functional group suitable for optical imaging, photoacoustic imaging, positron emission tomography (PET) imaging, single photon emission computed tomography (SPECT) imaging, or radiotherapy; and L a , L b , and L c are bifunctionalized linkers capable of forming chemical bonds with each other and with A, B, and C, respectively.

[0004] In another embodiment, the subject matter of the present disclosure has formula (II): [ka] providing a compound of the formula: wherein each y is independently an integer selected from the group consisting of 0, 1, and 2; R 1x , R 2x , and R 3x’ , are H, OH, halogen, and C, respectively. 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 independently selected from the group consisting of alkyl; R 4x is H, linear or branched C 1 - 6 Alkyl, -(CH 2 ) q4 -aryl and hydroxyl substituted straight or branched chain C 1 - 6 alkyl, wherein q4 is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; R 5x and R 6x are each independently H, a halogen, or -O-(CH 2 ) z3 -X a z1 is an integer from 0 to 4, z2 is an integer from 0 to 2, z3 is an integer from 1 to 6, and X a is a halogen, where R 5x and R 6xcannot both be H at the same time; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0005] In another aspect, the presently disclosed subject matter provides a compound of formula (III): In other embodiments, the subject matter of the present disclosure has formula (III): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); A and B can be the same or different, when B is absent or is a targeting moiety for FAP-α; C 1 may be absent or present, and when present is a chelating group; C 2 is a prosthetic group; L a and L b are each other as well as A, B, C 1 and C 2 is a bifunctional linker capable of forming a chemical bond with; L c1 and L c2 are independent of each other and A, B, L a , and L b is a bifunctional linker capable of forming a chemical bond with; C 1 If absent, L c1 is also absent, and if B is absent, then L b is also non-existent; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0006] In another embodiment, the subject matter of the present disclosure has formula (IV): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is a targeting moiety for prostate-specific membrane antigen (PSMA); C is any optically or radiolabeled functional group suitable for optical imaging, photoacoustic imaging, positron emission tomography (PET) imaging, single photon emission computed tomography (SPECT) imaging, or radiotherapy; and L a , L b , and L c are bifunctional linkers each capable of forming chemical bonds with each other and with A, B, and C, respectively; and and their stereoisomers and pharma- ceutically acceptable salts.

[0007] In other embodiments, the presently disclosed subject matter provides pharmaceutical compositions comprising a compound of formula (I-IV). In some embodiments, the pharmaceutical composition comprises one or more of a pharma- ceutically acceptable carrier, diluent, excipient, or adjuvant.

[0008] In another aspect, the subject matter of the present disclosure provides a method for imaging a disease or disorder associated with fibroblast activation protein-alpha (FAP-alpha) and / or prostate-specific membrane antigen (PSMA), the method comprising administering a compound of formula (I-IV), or a pharmaceutical composition thereof, wherein the compound of formula (I-IV) comprises an optically or radioactively labeled functional group suitable for optical imaging, photoacoustic imaging, PET imaging, or SPECT imaging; and obtaining an image.

[0009] In other aspects, the presently disclosed subject matter provides a method for inhibiting fibroblast activation protein-alpha (FAP-alpha) and / or prostate-specific membrane antigen (PSMA), the method comprising administering to a subject in need thereof an effective amount of a compound of Formula (I-IV), or a pharmaceutical composition thereof.

[0010] In another aspect, the presently disclosed subject matter provides a method for treating a fibroblast activation protein-alpha (FAP-α)- and / or prostate-specific membrane antigen (PSMA)-related disease or disorder, comprising administering to a subject in need of such treatment an effective amount of a compound of formula (I-IV), or a pharmaceutical composition thereof, wherein the compound of formula (I-IV) comprises a radiolabel functional group suitable for radiation therapy.

[0011] In some embodiments, the (FAP-α)-related disease or disorder is selected from the group consisting of proliferative diseases; diseases characterized by tissue remodeling and / or chronic inflammation; disorders associated with endocrine dysfunction; and blood clotting disorders.

[0012] In certain embodiments, the proliferative disease is selected from the group consisting of breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, renal cancer, lung cancer, melanoma, fibrosarcoma, bone and connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, and adenocarcinoma.

[0013] In certain embodiments, the prostate-specific membrane antigen (PSMA)-associated disease or disorder is selected from the group consisting of prostate cancer, kidney cancer, head cancer, neck cancer, head and neck cancer, lung cancer, breast cancer, prostate cancer, colon cancer, esophageal cancer, gastric cancer, leukemia / lymphoma, uterine cancer, skin cancer, endocrine cancer, urological cancer, pancreatic cancer, gastrointestinal cancer, ovarian cancer, cervical cancer, adenoma, and tumor neovasculature. In certain embodiments, the prostate-specific membrane antigen (PSMA)-associated disease or disorder comprises prostate cancer.

[0014] While certain aspects of the presently disclosed subject matter that are addressed in whole or in part by the presently disclosed subject matter have been described hereinabove, other aspects will become apparent as the description proceeds, as best described herein below, when taken in conjunction with the accompanying examples and figures. [Brief description of the drawings]

[0015] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0016] Having thus described the subject matter of the present disclosure in general terms, reference will now be made to the accompanying drawings, which are not necessarily drawn to scale:

[0017] [Figure 1] Representative compounds of formula (I), for example, are the chemical structures of [M]-SB-FAP-01, where M is 68Ga, 225Ac, 177Lu, and 64Cu, and have dual modalities against PSMA and FAP; [Diagram 2] 1 is a scheme for the synthesis of FAP-acetylene 1; [Diagram 3] 1 is a scheme for the synthesis of SB-FAP-01; [Figure 4] HPLC report for SB-FAP-01; [Diagram 5] This is the HRMS report for SB-FAP-01; [Figure 6] HPLC chromatogram of 68Ga-SB-FAP-01; showing the radioactive peak at 31.1 min (bottom) and the UV peak (λ = 254 nm) at 29.9 min associated with the unbound ligand SB-FAP-01 (top); [Figure 7] We demonstrate that the compound SB-FAP-01, a heterobivalent 68Ga-labeled contrast agent for positron emission tomography (PET), is capable of imaging tumors derived from FAP+HT-1080 human fibrotic cells engineered to overexpress FAP, but not wild-type tumors. Small animal PET / CT imaging was performed 60 min after intravenous administration in this mirroring experimental model; and [Figure 8] Cell binding data for SB-FAP-01 in PSMA+PIP cells (blue) and PSMA-flu cells (red) (uptake calculated as percent of incubated dose (ID%) per million cells). [Figure 9] Figures 9A and 9B show the structures of clinically relevant FAP-targeting scaffolds (Figure 9A) and 64Cu-FP-L1 and 64Cu-FP-L2 (Figure 9B); [Figure 10] Figures 10A and 10B show (Figure 10A) in celluro binding specificity of 64Cu-FP-L1 in U87, SK-MEL-24, and isogenic PSMA+ / PSMA- PC3 PIP / flu, PSMA+ / PSMA-786-O cells (mean ± SD, n=3) at 37 C. Data were obtained in two independent experiments with data shown from one. Receptor blocking studies were performed by co-incubating either 10 μM FAPI-04 (for FAP blocking) or 10 μM ZJ43 (for PSMA blocking) to assess binding specificity; (FIG. 10B) Cell surface FAP and PSMA expression by antibody-based flow cytometry was illustrated by the percentage of positive FAP and / or PSMA expressing cells for the following lines: human U87 glioma (no staining for PSMA, high staining for FAP); human SK-MEL-24 melanoma (moderate staining for PSMA, high staining for FAP); PC3 PIP (high staining for PSMA, no staining for FAP); PC3 flu (no staining for PSMA, no staining for FAP), PSMA+786-O (high staining for PSMA, low / no staining for FAP), 786-O vector (no staining for PSMA, no staining for FAP). Data were acquired in three independent experiments with data shown from one; [Figure 11]Figures 11A, 11B, 11C, 11D, 11E, and 11F show: Figure 11A. Experiment 1: Scheme; Figure 11B, 11C. Quantitative PET / CT imaging and region of interest (ROI) analysis of 64Cu-FP-L1 (7.4MBq in 150 μL saline) in U87 tumor-bearing mice (n=3 / time point). Figure 11D. Biodistribution data are shown as percentage of injected dose per gram of tissue (%ID / g), mean ± SD; Figure 11E. In vivo specificity with either FAPI-04 (FAP blocking) or ZJ43 (PSMA blocking) at 10 nmol per mouse, co-injected, 0.74 MBq in 150 μL saline (n=4); Figure 11F. Head-to-head comparison of H&E and IHC (10× magnification) of U87 tumor sections from the same cohort of mice displayed high FAP (brown staining) and PSMA (no staining) expression, with intermediate FAP and high PSMA expression in PSMA+PC3 PIP tumors. Significance was determined by unpaired t-test; [Figure 12] 12A, 12B, 12C, 12D, 12E, and 12F show: 12A. Experiment 2: Schematic; 12B. Quantitative PET imaging and region of interest (ROI) analysis of 64Cu-FAP-L1 in SK-MEL-24 tumor-bearing mice (n=4 / group). Tumor uptake is indicated by black arrows; 12C. Receptor blockade: tumor (red), kidney (yellow) dotted area [co-injection of PSMA-targeted ZJ43, or co-injection of FAP-targeted FAPI-04 or autoblockade with FP-L1 (10 nmol per mouse) was performed at 30 min]. The absence of kidney uptake upon administration of ZJ43 or non-radiolabeled FP-L1 indicates PSMA binding specificity; 12D. Quantitative depiction of PET / CT data; 12E. Mice were euthanized for biodistribution study 2 h after imaging. Biodistribution data are presented as percentage of injected dose per gram of tissue (%ID / g), mean ± SD. Figure 12F. H&E (10x magnification) for morphology and brown staining (IHC) for FAP and PSMA expression from the same cohort of mice used in this experiment showing high FAP expression and no PSMA expression; [Figure 13]Figures 13A, 13B, 13C, 13D, and 13E show whole-body PET imaging of KPC mice with 64Cu-FP-L1 showing localization in pancreatic lesions. Figure 13A. Experiment 3: Schematic; Figure 13B. Age-matched healthy littermates (left) and KPC mice (right) after 2 hours. Focused uptake in the abdomen (red arrow), kidney (yellow dotted area), and salivary gland (red circle); Figure 13C. (Left) Ex vivo near-infrared fluorescence imaging of selected tissues 2 hours after IRDye800-FP-L1 administration showing focal uptake in the pancreas (yellow dotted area), metastatic lesions (yellow arrows), and (right) white light photographs of tissues; Figure 13D. From left to right: H&E (whole tissue and 40x magnification) obtained demonstrating the presence of PDAC and PanIN lesions; FAP-positive IHC of PDAC (10x magnification); and FAP-negative healthy tissue. Figure 13E. PSMA-specific staining of healthy pancreas and kidney as well as tumor tissue sections; [Figure 14] Figure 14A, Figure 14B, and Figure 14C show: Figure 14A. PET / CT imaging of 64Cu-FP-L1 in PSMA+786-O (n=2) tumor-bearing mice shows focal uptake in the dotted areas of the tumor (red) and kidney (yellow); Figure 14B. Biodistribution data shown as percentage of injected dose per gram of tissue (%ID / g), mean ± SD (n=4); Figure 14C. Left to right: H&E and IHC micrographs of tumor tissue from the same cohort of mice, 10x magnification, showing clear cell morphology (10x magnification) and positive (brown) staining for FAP and PSMA expression; [Figure 15] 1 is a schematic diagram of the synthesis scheme of FP-L1; [Figure 16] 1 is a schematic diagram of the synthesis scheme of FP-L2; [Figure 17] Dose-dependent responses (sigmoidal curves) of FP-L1 and FPL2 to the indicated substrates PSMA (NAAG), FAP and PREP (Z-Gly-Pro-AMC) and DPPIV (H-Gly-Pro-AMC) are shown in the presence of either the corresponding recombinant enzymes for FAP, PREP and DPPIV, or LNCaP cell lysate for PSMA; [Figure 18] Static PET imaging of U87 tumor-bearing mice during 1-4 hours after 64Cu-FP-L1 administration. Blocking studies using co-injection of 50 nmol FAPI-04 were also performed, showing a significant reduction in tumor and non-specific tissue uptake. This study was performed from a batch of mice bearing small tumors that underwent biodistribution studies at 2 hours; [Figure 19] Sequential PET imaging and biodistribution studies of 64Cu-FPL2 at 24 hours are presented; [Figure 20] Shows the synthesis of IR800Dye-FP-L1; and [Figure 21] H&E staining of the lungs, liver and gallbladder of KPC mice is shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0018] Detailed Description

[0019] The subject matter of the present disclosure will now be described in more detail below with reference to the accompanying figures, in which some, but not all, embodiments of the present disclosure are shown. Like numbers refer to like elements throughout. The subject matter of the present disclosure may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the subject matter of the present disclosure described herein will readily come to mind to one skilled in the art to which the subject matter of the present disclosure pertains having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. It should therefore be understood that the subject matter of the present disclosure is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0020] I. Heterobivalent and homobivalent agents targeting fibroblast activation protein alpha and / or prostate-specific membrane antigen

[0021] Fibroblast activation protein-α (FAP-α) is a type II integral membrane serine protease of the prolyl oligopeptidase family, which are distinguished by their ability to cleave Pro-AA peptide bonds (where AA represents any amino acid). FAP-α exists as a homodimer to perform its enzymatic function. It has been shown to play a role in cancer by modifying bioactive signal peptides through this enzymatic activity (Kelly et al., 2005; Edosada et al., 2006). Expression of FAP-α has been detected on the surface of fibroblasts in epithelial cancers, including but not limited to malignant breast, colorectal, skin, prostate, pancreatic cancer, and the like, and in the stroma surrounding more than 90% of inflammatory diseases, including but not limited to arthritis, fibrosis, and the like, and is rarely expressed in healthy tissues. Inhibitors that selectively target FAP-α have been reported (Lo et al., 2009; Tsai et al., 2010; Ryabtsova et al., 2012; Poplawski et al., 2013; Jansen et al., 2014).

[0022] More specifically, FAP-α expression has been detected on the surface of fibroblasts in the stroma surrounding >90% of epithelial cancers investigated, including malignant breast, colorectal, skin, prostate, and pancreatic cancers (Garin-Chesa et al., 1990; Rettig et al., 1993; Tuxhorn et al., 2002; Scanlan et al., 1994). It is a characteristic marker of carcinoma-associated fibroblasts (CAFs), where it plays an important role in promoting angiogenesis, proliferation, invasion, and suppressing tumor cell death (Allinen et al., 2004; Franco et al., 2010). In healthy adult tissues, FAP-α expression is restricted to areas of tissue remodeling or wound healing. (Scanlan et al., 1994; Yu et al., 2010; Bae et al., 2008; Kraman et al., 2010). In addition, FAP-α positive cells are observed in areas of chronic inflammation, arthritis, and fibrosis during embryonic development, and in soft tissue and bone sarcomas. (Scanlan et al., 1994; Yu et al., 2010). These features make FAP-α an imaging and radiotherapy target for cancer and inflammatory diseases.

[0023] Because FAP-α is expressed in the tumor stroma, anti-FAP antibodies have been investigated for radioimmunotargeting of malignant tumors, including murine F19, sibrotuzumab (a humanized version of the F19 antibody), ESC11, ESC14, and others (Welt et al., 1994; Scott et al., 2003; Fischer et al., 2012). Antibodies have also demonstrated feasibility for imaging inflammation, such as rheumatoid arthritis (Laverman et al., 2015). However, the use of antibodies as molecular imaging agents suffers from pharmacokinetic limitations, including slow blood and non-target tissue clearance (usually 2-5 days or longer) and non-specific organ uptake. Low molecular weight (LMW) agents demonstrate fast pharmacokinetics after administration and high specific signal within a clinically convenient time frame. Also, they can be more easily synthesized in radiolabeled form and the path to regulatory approval may be shorter (Coenen et al., 2010; Coenen et al., 2012; Reilly et al., 2015). However, to date, no LMW ligands with ideal properties for nuclear imaging of FAP-α have been reported.

[0024] Similarly, prostate-specific membrane antigen (PSMA) is a type II integral membrane protein expressed on the surface of prostate tumors, particularly in castration-resistant, advanced, and metastatic disease (Huang, 2004; Schuelke, 2003). PSMA is also expressed on the neovascular endothelium of most solid tumors, such as lung, colon, pancreatic, renal carcinoma, and cutaneous melanoma, but not on normal vasculature (Liu, 1997; Chang, 1999), making it an excellent target for imaging and targeted therapy of these cancers.

[0025] The subject matter of the present disclosure provides, in part, compounds that include FAP-α and PSMA-selective targeting moieties that can be modified with optical dyes, radiometal chelation complexes, and other radiolabeled prosthetic groups, thus providing a platform for imaging and radiotherapy targeted to FAP-α and PSMA.

[0026] Radionuclide molecular imaging, including positron emission tomography (PET), is the most mature molecular imaging technique without the limitation of tissue penetration. Due to the benefits of its high sensitivity and quantifiability, radionuclide molecular imaging plays an important role in clinical and preclinical research (Youn et al., 2012; Chen et al., 2014). Many radionuclides, mainly beta- and alpha-emitters, have been investigated for targeted immunotherapy, and include both radioactive halogens and radioactive metals (see Table 1 for representative therapeutic radionuclides).

[0027] [Table 1]

[0028] Radioisotopes suitable for use with the presently disclosed subject matter also include, but are not limited to, 11 C. 18 F, 51 Cr, 68 Ga, 99m Tc, 130 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 88 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 72 As,72 Se, 97 Ru, 109 Pd, 105 Rh, 101 mRh, 119 Sb, 128 Ba, 124 I, 197 Hg, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 64 Cu, 198 Au, 225 Ac, 227 Th, and 199 Contains Ag.

[0029] Fluorescent dyes suitable for use with the presently disclosed subject matter include, but are not limited to, xanthenes, acridines, oxazines, cyanines, styryl dyes, coumarins, porphines, fluorescent proteins, perylenes, boron-dipyrromethenes, and phthalocyanines.

[0030] The highly potent and specific binding moiety targeting FAP-α and PSMA allows its use in nuclear imaging and radiotherapy. The subject matter of the present disclosure provides the first synthesis of a nuclear imaging and radiotherapy agent based on this dual targeting moiety against FAP-α and PSMA.

[0031] Thus, in some embodiments, the presently disclosed subject matter provides potent and selective low molecular weight (LMW) ligands of FAP-α, i.e., FAP-α and PSMA selective inhibitors conjugated with optical dyes or radiolabeling groups, including metal chelators and metal complexes, enabling in vivo optical imaging, nuclear imaging (optical, PET and SPECT), and radiation therapy targeted to FAP-α and PSMA.

[0032] Importantly, the compounds of the present disclosure can be modified, for example, conjugated with labeling groups without significant loss of their potency. The approach of the present disclosure can be used to conjugate, for example, but not limited to, optical dyes and PET or SPECT isotopes for FAP-α and / or PSMA related imaging applications. 68 Ga, 64 Cu, 18 F, 86 Y, 90 Y, 89 Zr, 111 In, 99m Tc, 125 I, 124 The disclosed approach can be conveniently labeled with radiotherapeutic isotopes, including, but not limited to, for FAP-α and / or PSMA-related radiotherapy. 90 Y, 177 Lu, 125 I, 131 I, 211 At, 111 In, 153 Sm, 186 Re, 188 Re, 67 Cu, 212 Pb, 225 Ac, 213 Bi, 212 Bi, 212 Pb, and 67 Targeting compounds with Ga inclusions allow for radiolabeling of FAP-α and PSMA.

[0033] A. Compound

[0034] The subject matter of the present disclosure provides heterobivalent compounds that target prostate-specific membrane antigen (PSMA) and fibroblast activation protein (FAP), which can serve as a platform for cancer imaging and treatment.Imaging and treatment agents have been developed to target each of these cancer-related proteins independently.Without wishing to be bound by a particular theory, it is believed that it was necessary to combine agents that target PSMA and FAP into one platform, because they depict different aspects of tumors and their microenvironments.

[0035] The imaging aspect can include various modalities including, for example, near-infrared optical imaging, positron emission tomography (PET), single photon emission computed tomography (SPECT), and magnetic resonance imaging (MRI).

[0036] Therapeutic aspects can include a variety of therapeutic nuclides, including, but not limited to, 177 Lu, 211 At, 225 Chemical toxins can also be affixed to the platform.

[0037] In some embodiments, the subject matter of the present disclosure has formula (I): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is a targeting moiety for prostate-specific membrane antigen (PSMA) or a targeting moiety for FAP-α, and when A and B are each a targeting moiety for FAP-α, they can be the same or different; C is any optically or radiolabeled functional group suitable for optical imaging, photoacoustic imaging, positron emission tomography (PET) imaging, single photon emission computed tomography (SPECT) imaging, or radiotherapy; and L a , L b , and L c are bifunctionalized linkers each capable of forming chemical bonds with each other and with A, B, and C, respectively.

[0038] Suitable FAP-α specific targeting moieties are provided in International PCT Patent Application Publication No. WO2019 / 083990 A2 to Yang et al., relating to Imaging and Radiotherapeutics Targeting Fibroblast Activation Protein-α (FAP-α), published May 2, 2019, which is incorporated by reference in its entirety.

[0039] In certain embodiments, the targeting moiety for fibroblast activation protein alpha (FAP-α) has the following structure: [ka] having; wherein each y is independently an integer selected from the group consisting of 0, 1, and 2; R 1x , R 2x , and R 3x’ , are H, OH, halogen, and C, respectively. 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 independently selected from the group consisting of alkyl; R 3x are H, -CN, -B(OH) 2 , -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O) 2 Aryl, -CO 2 H, -SO 3 H, -SO 2 NH 2 , -PO 3 H2 and 5-tetrazolyl; R 4x is H, linear or branched C 1 - 6 Alkyl, -(CH 2 ) q4 -aryl and hydroxyl substituted straight or branched chain C 1 - 6 alkyl, wherein q4 is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; R 5x , R 6x , and R 7x are H, -OH, oxo, halogen, and -C 1-6 Alkyl, -OC 1-6 Alkyl, -SC 1-6 Alkyl, -NR 8x R 9x , -OR 12x , -Het 2 and -Ar 2 Each C 1-6 The alkyl is optionally substituted with 1 to 3 substituents selected from -OH and halogen; R 8x , R 9x , and R 12x are H, -OH, halo, and -C 1-6 Alkyl, -OC 1-6 Alkyl, -SC 1-6 Alkyl, and -Ar 3 independently selected from the group consisting of; R 10x , R 11x , R 13x and R 14x are H, -OH, halogen, and -C, respectively. 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 Ar is independently selected from the group consisting of alkyl; 1 , Ar 2 and Ar 3each independently is a 5- or 6-membered aromatic monocycle optionally containing 1 or 2 heteroatoms selected from O, N and S; 1 , Ar 2 and Ar 3 -NR 10x R 11x , -C 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 Optionally and independently substituted with 1 to 3 substituents selected from alkyl; Het 2 is a 5- or 6-membered non-aromatic monocyclic ring optionally containing one or two heteroatoms selected from O, N and S; Het 2 is optional -NR 13x R 14x , -C 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 substituted with 1 to 3 substituents selected from alkyl; v is 0, 1, 2, or 3; and [ka] represents a 5 to 10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, said heterocycle optionally further containing 1, 2 or 3 heteroatoms selected from O, N and S; So [ka] is the linker, L 3 indicates the point of attachment of the FAP-α binding ligand to, where the point of attachment can be through any of the carbon atoms of, their 5 to 10 membered, N-containing aromatic or non-aromatic monocyclic or bicyclic heterocyclic rings; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0040] In some embodiments, the linker between the 5 to 10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle and the pyrrolidine ring is derived from an amino acid, e.g., glycine, alanine, phenylalanine, valine, serine, and threonine: [ka] It is.

[0041] In certain embodiments, R 4x is H (glycine), -CH 3 (Alanine), -CH 2 -phenyl(phenylalanine), -CH(CH 3 ) 2 (valine), -CH 2 -OH (serine), and -CH(OH)CH 3 (Threonine).

[0042] In certain embodiments, [ka] is the following: [ka] Selected from the group consisting of; In the formula, * represents -(CH 2 ) v - denotes the point of attachment of a 5 to 10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle to the aryl group.

[0043] In further certain embodiments, A is: [ka] or wherein A and B are an FAP-α targeting moiety having the structure: During the ceremony [ka] is the linker, L 3 where the attachment point can be at any carbon atom 5, 6, 7, or up to 8 of the quinolinyl ring thereof; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0044] In still further embodiments, A is: [ka] or A and B are each independently selected from the group consisting of:

[0045] In still further certain embodiments, A is: [ka] and stereoisomers thereof, or A and B are each independently selected from the group consisting of:

[0046] In still further certain embodiments, A is: [ka] or A and B are each independently selected from the group consisting of:

[0047] Suitable FAP inhibitors are disclosed in International PCT Patent Application Publication No. WO2019 / 154886 to Haberkorn et al. for FAP Inhibitors, published on August 15, 2019, which is incorporated by reference in its entirety. Exemplary FAP ligands, linkers, and reporting moieties include those having the following formula (I): [ka] Contains compounds; During the ceremony: Q, R, U, V, W, Y, Z are individually present or absent under the proviso that at least three of Q, R, U, V, W, Y, Z are present; Q, R, U, V, W, Y, Z are O, CH 2 , N.R. 4 , C=O, C=S, C=NR 4 , H.C.R. 4 and R 4 CR 4 with the proviso that the two oxygen atoms are not directly adjacent to each other; R 1 and R 2 Regardless of -H, -OH, halo, C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl; R 3 -H, -CN, -B(OH) 2 , -C(O)-alkyl, -C(O)-aryl-, -C=CC(O)-aryl, -C=CS(O) 2 -aryl, -CO 2 H, -SO 3 H, -SO 2 H 2 ,-PO 3 H 2 and 5-tetrazolyl; R 4 -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl, and -C 1-6 -aralkyl, each of said -C 1-6 - alkyl is optionally substituted with 1 to 3 substituents selected from -OH, oxo, halo, and optionally connected to Q, R, U, V, W, Y, or Z; R 5 -H, halo, and C 1-6 -alkyl; R 6 , and R 7 Regardless of -H, [ka] From the group consisting of R 6 and R 7 is selected under the proviso that: D is a linker; A is NR 4 , O, S, and C-H 2 Selected from the group consisting of; E is C 1-6 Alkyl, [ka] selected from the group consisting of; and where i is 1, 2, or 3; where j is 1, 2, or 3; where k is 1, 2, or 3; wherein m is 1, 2, or 3; B is S, NR 4 , N.R. 4 -O, NR 4 -C 1-6 -Alkyl, NR 4 -C 1-6 -Alkyl-NR 4 and 5 to 10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, preferably further containing one or two heteroatoms selected from O, N, and S, preferably further containing one or two nitrogen atoms, and preferably where NR 4 -C 1-6 -Alkyl-NR 4 and N-containing heterocycles are C 1-6 -Alkyl, aryl, C1-6 -substituted with 1 to 3 substituents selected from the group consisting of aralkyl; and R 8 is selected from the group consisting of a radioactive moiety, a chelating agent, a fluorescent dye, a contrast agent, and combinations thereof; [ka] is an l-naphthyl moiety or a 5 to 10 membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, in which there are two ring atoms between the N atom and X; said heterocycle optionally further contains 1, 2 or 3 heteroatoms selected from O, N and S; and X is a C atom; or a pharma- ceutically acceptable tautomer, racemate, hydrate, solvate, or salt thereof.

[0048] In certain embodiments, A is a FAP-α targeting moiety or one of the following: [ka] is a ligand having the structure: X in the formula 1 and X 2 are each independently H or F.

[0049] In certain embodiments, the FAP inhibitor disclosed in WO2019 / 154886 is a compound, including a FAP ligand, a linker, and a reporting moiety, disclosed in one or more of Tables 1, 2, 3, 4, and 5, or any compound disclosed on page 44, line 1 to page 75, line 6, which are incorporated herein by reference, including, but not limited to, FAPI-1, FAPI-2, FAPI-3, FAPI-4, FAPI-5, FAPI-6, FAPI-7, FAPI-8, FAPI-9, FAPI-10, FAPI-11, FAPI-12, FAPI-13, FAPI-14, FAPI-15, FAPI-16, FAPI-17, FAPI-18, FAPI-19, FAPI-20, FAPI-21, FAPI-22, FAPI-23, FAPI-24, FAPI-25, FAPI-26, FAPI-27, FAPI-28, FAPI-29, FAPI-30, FAPI-31, FAPI-32, FAPI-33, FAPI-34, FAPI-35, FAPI-36, FAPI-37, FAPI-38, FAPI-39, FAPI-40, FAPI-41, FAPI-42, FAPI-43, FAPI-44, FAPI-45, FAPI-46, FAPI-47, FAPI-48, FAPI-49 ...9, FAPI-49, FAPI-49, FAPI-49, FAPI-49, FAPI-49, F PI-5, FAPI-6, FAPI-7, FAPI-8, FAPI-9, FAPI-10, FAPI-11, FAPI-12, FAPI-13, FAPI-14, FAPI-15, FAPI-16, FAPI-17, FAPI-1 8, FAPI-19, FAPI-20, FAPI-21, FAPI-22, FAPI-23, FAPI-24, FAPI-25, FAPI-26, FAPI-27, FAPI-28, FAPI-29, FAPI-30, FAPI-3 1, FAPI-32, FAPI-33, FAPI-34, FAPI-35, FAPI-36, FAPI-37, FAPI-38, FAPI-39, FAPI-40, FAPI-41, FAPI-42, FAPI-43, FAPI- 44, FAPI-45, FAPI-46, FAPI-47, FAPI-48, FAPI-49, FAPI-50, FAPI-51, FAPI-52, FAPI-53, FAPI-54, FAPI-55, FAPI-56, FAPI FAPI-57, FAPI-58, FAPI-60, FAPI-61, FAPI-62, FAPI-63, FAPI-64, FAPI-65, FAPI-66, FAPI-67, FAPI-68, FAPI-69, FAPI-70, FAPI-71, FAPI-72, FAPI-73, FAPI-74, FAPI-75, FAPI-76, FAPI-77, FAPI-78, and FAPI-79, each of which is incorporated herein by reference.

[0050] In some embodiments, the FAP-α ligand includes the following: Jansen et al., Selective Inhibitors of Fibroblast Activation Protein (FAP) with a (4-Quinolinoyl)-glycyl-2-cyanopyrrolidine Scaffold. ACS Med Chem Lett. 2013 Mar 18;4(5):491-6; Jansen et al., Extended structure-activity relationship and pharmacokinetic investigation of (4-quinolinoyl)glycyl-2-cyanopyrrolidine inhibitors of fibroblast activation protein (FAP). J Med Chem. (Journal of Medicinal Chemistry) 2014 Apr 10;57(7):3053-74, each of which is incorporated by reference in its entirety. Such FAP-α ligands include the substituted (4-quinolinoyl)-glycyl-2-cyanopyrrolidine scaffolds disclosed therein: [ka] Includes; During the ceremony: X 1 and X 2 are each independently H or F; and R 15x is H, C 1-6 Alkyl, halogen, trihalomethoxyl, C 1-6 alkoxyl, and 4-methoxyphenyl.

[0051] Also included are the FAP ligands disclosed in Roy et al., Design and validation of fibroblast activation protein alpha targeted imaging and therapeutic agents, Theranostics 2020, 10(13), 5778-5789, which are incorporated herein by reference in their entireties, including, but not limited to, the following: [ka] Includes:

[0052] In certain embodiments, A has the structure: [ka] or A and B each are.

[0053] In other embodiments, B has the structure: [ka] is a targeting moiety for FAP-α having the formula:

[0054] In certain embodiments, B has the structure: [ka] a PSMA targeting moiety having the formula: During the ceremony: Z is tetrazole or CO 2 Q; Q is H or a protecting group; q1 is an integer selected from the group consisting of 1, 2, 3, 4, and 5; q 2 is an integer from selected from 0 or 1; and R y is independently H or -(CH 2 ) q3 -R y1 Where q 3 is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6, and R y1 is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline;

[0055] In further certain embodiments, B has the structure: [ka] is a PSMA targeting moiety having the formula:

[0056] In certain embodiments, R y1 is the following: [ka] Selected from the group consisting of; In the formula, X is Br, 75 Br, 76 Br, 77 Br, 80m Br, 82 Br, I, 124 I, 123 I, 125 I, 131 I, At, and 211 At.

[0057] Suitable linkers are disclosed in U.S. Patent Application Publication No. US2011 / 0064657 A1, published March 17, 2011 to Pomper et al., for "Labeled Inhibitors of Prostate Specific Membrane Antigen (PSMA), Biological Evaluation, and Use as Imaging Agents," and U.S. Patent Application Publication No. US2012 / 0009121 A1, published January 12, 2012 to Pomper et al., for "PSMA-Targeting Compounds and Uses Thereof," each of which is incorporated by reference in its entirety.

[0058] In certain embodiments, L a , L b , and L c are each independently selected from the group consisting of (a), (b), (c), or (d): (a) [ka] and; During the ceremony: p 1 , p 2 , p 3 and p 4 may be in any order; t 1 and t 2 are each an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; p 1、 p 3 , and p 4 are each independently 0 or 1; p 2 is an integer selected from the group consisting of 0, 1, 2, and 3, and p 2 When is 2 or 3, each R 1are the same or different; m 1 and m 2 are each an integer independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; W 1 is selected from the group consisting of a bond, -S-, -C(=O)-, -C(=O)-NR-, and -NR-C(=O)-; W 2 is a bond, -S-, -CH 2 -C(=O)-NR-, -C(=O)-, -NRC(=O)-, -NR'C(=O)NR-, -NRC(=S)NR' 2 -, -NRC(=O)O-, -OC(=O)NR-, -OC(=O)-, -C(=O)NR-, -NR-C(=O)-, -C(=O)O-, -(O-CH 2 -CH 2 ) q - and -(CH 2 -CH 2 -O) q wherein q is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; Each R or R' is independently H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and -OR 4 where R 4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl, where q is defined as immediately above; Tz is a triazole group which may be present or absent, and when present, [ka] Selected from the group consisting of; Each R 1 Regardless of H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, -(CH 2) q -C 3 -C 12 Aryl, -C 4 -C 16 Alkylaryl, or -(CH 2 ) q -C 4 -C 16 R is alkylaryl; 2 and R 3 are H and -CO independently 2 R 5 where R 5 is H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, and C 4 -C 16 alkylaryl, where R 2 or R 3 One of them is CO 2 R 5 If , then else H; V is selected from the group consisting of -C(O)-, -C(S)-, -NRC(O)-, -NRC(S)-, and -OC(O)-; (b) [ka] and In the formula, p 1、 p 2 , p 3 , m 1 , m 2 , Tz, W 2、 R, R 1 , R 2 , R 3 and V is defined above; (c)-L 1 -, -L 2 -L 3 - or -L 1 -L 2 -L 3 - in which: L 1 -NR-(CH 2 ) q -[O-CH2 -CH 2 -O] q -(CH 2 ) q -C(=O)-; L 2 -NR-(CH 2 ) q -C(COOR 5 )-NR-; and L 3 is -(O=)C-(CH 2 ) q -C(=O)-; wherein each q is independently an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as defined above; and (d)-(CR 6 H) q -(CH 2 ) q -C(=O)-NR-(CH 2 ) q -O- or -NR-(CH 2 ) q -O-; where each q and R are defined above; and R 6 H or -COOR 5 It is.

[0059] Further, in certain embodiments, one or more of L a , L b , and L c is the following: [ka] where u is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as set out above.

[0060] In certain embodiments, C is 18 F, 124 I, 125 I, 131 I, and 211A radiolabeled prosthetic group containing a radioisotope selected from the group consisting of At.

[0061] In further embodiments, the radiolabeled prosthetic group is: [ka] Selected from the group consisting of; In the formula, each X is independently 18 F, 124 I, 125 I, 131 I, and 211 At; each R and R' is defined above; and each n is independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0062] In still further embodiments, the radiolabeled prosthetic group is: [ka] is selected from the group consisting of:

[0063] In certain embodiments, C comprises a chelating agent. In further certain embodiments, the chelating agent is selected from the group consisting of DOTAGA (1,4,7,10-tetraazacyclododecane, 1-(glutaric acid)-4,7,10-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTASA (1,4,7,10-tetraazacyclododecane-1-(2-succinic acid)-4,7,10-triacetic acid), CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), DEPA (7-[2-(bis-carboxymethylamino)-ethyl]-4,10-bis-carboxymethyl-1,4,7,10-tetraaza-cyclod odec-1-yl-acetic acid), 3p-C-DEPA (2-[(carboxymethyl)][5-(4-nitrophenyl-1-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]pentan-2-yl)amino]acetic acid), TCMC (2-(4-isothiocyanotobenzyl)-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamonylmethyl)-cyclododecane), oxo-DO3A (1-oxa-4,7,10-triazacyclododecane-5-S-(4-isothiocyanatobenzyl)-4,7,10-triacetic acid), p-NH 2-Bn-oxo-DO3A (1-oxa-4,7,10-tetraazacyclododecane-5-S-(4-aminobenzyl)-4,7,10-triacetic acid), TE2A ((1,8-N,N′-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane), MM-TE2A, DM-TE2A, CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane), CB-TE1A1P (4,8,11-tetraazacyclotetradecane-1-(methyl)- CB-TE2P (1,4,8,11-tetraazacyclotetradecane-1,8-bis(methanephosphonic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N′,N″-triacetic acid), NODA (1,4,7-triazacyclononane-1,4-diacetate); NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), (NOTAGA) 1,4,7-Triazonan-1,4-diyl)diacetic acid DFO (deferoxamine), NETA ([4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid), TACN-TM (N,N',N'',tris(2-mercaptoethyl)-1,4,7-triazacyclononane), Diamsar (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo(6,6,6)eicosapentaenoic acid), and n-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), Sarar (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), AmBaSar (4-((8-amino-3,6,10,13,16,19-hexazabicyclo[6.6.6]icosan-1-ylamino)methyl)benzoic acid), and BaBaSar.

[0064] In still further embodiments, the chelating agent is: [ka] [ka] [ka] [ka] [ka] is selected from the group consisting of:

[0065] One of ordinary skill in the art will recognize that commercially available chelating agents may include activating agents, such as agents capable of reacting with primary amines. Such agents include, but are not limited to, N-hydroxysuccinimide (NHS), N-hydroxysulfosuccinimide (sulfo-NHS), anhydrides, maleimides, N-benzyl, 4-isothiocyanatobenzyl (p-NCS-Bz), NH 2 -MPAA, Propargyl, TA, N-(2-aminoethyl)ethanamide, NH 2 -PEG4, and a hydrophilic dPEG spacer linked to a tetrafluorophenyl (TFP) ester. These agents can be attached to or form part of a linker that is attached to a chelating agent.

[0066] In certain embodiments, the chelating agent further comprises a radioactive metal. 60 Cu, 62 Cu, 64 Cu, 67 Cu, 203 Pb, 212 Pb, 225 Ac, 177 Lu, 99m Tc, 68 Ga, 149 Tb, 86 Y, 90 Y, 111 In,186 Re, 188 Re, 153 Sm, 89 Zr, 213 Bi, 212 Bi, 212 Pb, 67 Ga, 47 Sc, and 166 Ho.

[0067] In certain embodiments, C comprises an optical dye. In further certain embodiments, the optical dye comprises a fluorescent dye. In yet further certain embodiments, the fluorescent dye comprises a fluorescent dye that emits in the near infrared spectral region. In certain embodiments, the fluorescent dye is selected from the group consisting of polymethine dyes, coumarin dyes, xanthene dyes, and boron-dipyrromethene (BODIPY) dyes.

[0068] In certain embodiments, the polymethine dye is selected from the group consisting of carbocyanine dyes, indocarbocyanine dyes, oxacarbocyanine dyes, thiacarbocyanine dyes, and merocyanine dyes. In certain embodiments, the xanthene dye is selected from the group consisting of fluorescein dyes and coumarin dyes. In particular embodiments, the fluorescent dye is one of the following: BODIPY FL, BODIPY R6G, BODIPY TR, BODIPY TMR, BODIPY 493 / 503, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, and BODIPY 650 / 665; Cy3, Cy3.5, Cy5, Cy5.5, Cy7, and Cy7.5; VivoTag-645, VivoTag-680, VivoTag-S680, VivoTag-S750, VivoTag-800; Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680, Alexa Fluor 700, Alexa Fluor 750, and Alexa Fluor790; Dy677, Dy676, Dy682, Dy752, Dy780; DyLight 350, DyLight 405, DyLight 488, DyLight 547, DyLight 550, DyLight 594, DyLight 633, DyLight 647, DyLight 650, DyLight 680, DyLight 755, and DyLight 800; HiLyte Fluor 405, HiLyte Fluor 488, HiLyte Fluor 532, HiLyte Fluor 555, HiLyte TM Fluor 594, HiLyte Fluor 647, HiLyte Fluor 680, HiLyte Fluor 750; IR800 (dimethyl{4-[1,5,5-tris(4-dimethylaminophenyl)-2,4-pentadienylidene]-2,5-cyclohexadien-1-ylidene}ammonium perchlorate), IRDye 650, IRDye 680RD, IRDye 680LT, IRDye 700, IRDye 700DX, IRDye 750, IRDye 800, IRDye 800CW, IRDye 800RS; and Selected from the group consisting of ADS1065A, ADS1075A, ADS775MI, ADS775MP, ADS775PI, ADS775PP, ADS780HO, ADS780WS, ADS785WS, ADS790WS, ADS795WS, ADS798SM, ADS800AT, ADS815EI, ADS830AT, ADS830WS, ADS832WS, ADS845MC, and ADS920MC.

[0069] In a more particular embodiment, the optical dye is: [ka] [ka] is selected from the group consisting of:

[0070] Suitable fluorescent agents and linkers are disclosed in International PCT Patent Application No. WO2018232280 to Pomper et al., entitled "PSMA Targeted Fluorescent Agents for Image Guided Surgery," published on December 20, 2018, which is incorporated herein by reference in its entirety.

[0071] In some embodiments, the compounds of the present disclosure can be made by linking near-infrared, closed chain, sulfocyanine dye to prostate specific membrane antigen ligand.For example, the prostate specific membrane antigen ligand used in the compounds of the present disclosure is described in International PCT Patent Application Publication No. WO2010 / 108125 to Pomper et al., published on September 23, 2010, which is incorporated herein in its entirety.

[0072] Compounds can be assembled to form linkages such as ureas (-NRC(O)NR-), thioureas (-NRC(S)NR-), amides (-C(O)NR- or -NRC(O)-), or esters (-C(O)O- or -OC(O)-) by reaction between different components. Urea linkages can be easily prepared by reaction between amines and isocyanates, or between amines and activated carbonamides (-NRC(O)-). Thioureas can be easily prepared from reaction of amines with isothiocyanates. Amides (-C(O)NR- or -NRC(O)-) can be easily prepared by reaction between amines and activated carboxylic acids or esters such as acyl halides or N-hydroxysuccinimide esters. Carboxylic acids can also be activated in situ, for example with coupling reagents such as carbodiimides, or carbonyldiimidazole (CDI). Esters can be formed by the reaction between an alcohol and an activated carboxylic acid. Triazoles are readily prepared by the reaction between an azide and an alkyne, optionally in the presence of a copper (Cu) catalyst.

[0073] Prostate specific membrane antigen ligands can also be prepared by sequential addition of components, such as lysine-urea-glutamic acid compounds, to preformed urea as described by Banerjee et al. (J. Med. Chem. 51, 4504-4517, 2008). Other urea-based compounds may also be used as building blocks. Exemplary syntheses of the near-IR, closed-chain, sulfocyanine dyes used in the compositions of the present disclosure are described in U.S. Pat. Nos. 6,887,854 and 6,159,657, and are incorporated herein in their entireties. In addition, some of the IR, closed-chain, sulfocyanine dyes of the subject matter of the present disclosure can be synthesized using DyLight. TM800 (ThermoFisher).

[0074] In some embodiments, the disclosed compounds include a fluorescent dye and are used for photoacoustic imaging of tumors.See, for example, Zhang et al., Prostate-specific membrane antigen-targeted photoacoustic imaging of prostate cancer in vivo, J. of Biophotonics, 2018;11:e201800021. Thus, in certain embodiments, C includes a dye suitable for use in photoacoustic imaging.

[0075] In certain embodiments, C comprises a photosensitizer. Photodynamic therapy (PDT) is a minimally invasive cancer treatment and has been used in clinical practice to improve the quality of life and survival time of cancer patients. However, the lack of specific delivery of photosensitizer is a major limitation of PDT. In non-targeted, conventional photodynamic therapy, photosensitizer cannot be specifically delivered to tumors, and photosensitizer circulates in the body for a long time after treatment and often causes hypersensitivity to light for several months.

[0076] Urea-based photosensitizers target prostate-specific membrane antigen (PSMA) for imaging and targeted treatment of PSMA-expressing tumors and cancers and are disclosed in International PCT Patent Application No. WO2015057692, published April 23, 2015, to Pomper et al., relating to Prostate-Specific Membrane Antigen-Targeted Photosensitizers for Photodynamic Therapy, and U.S. Pat. No. 10,232,058, published March 19, 2019, to Pomper et al., relating to Prostate-Specific Membrane Antigen-Targeted Photosensitizers for Photodynamic Therapy, each of which is incorporated by reference in its entirety. [ka]

[0077] In certain embodiments, the compound of Formula (I) has the following structure: [ka] having; wherein b1, b2, b3, b4, b5, b6, and b7 are each independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8.

[0078] In certain embodiments, the compound of formula (I) has the following structure: [ka] has.

[0079] In a further particular embodiment, the compound of formula (I) is: [ka] and; In the formula, M is 68 Ga, 225 Ac, 177 Lu, and 64Cu.

[0080] In some embodiments, A and B are each: [ka] The FAP-α targeting moiety has the structure:

[0081] In certain embodiments, the compound is: [ka] is selected from the group consisting of:

[0082] In certain embodiments, the compound is: [ka] It is.

[0083] In other embodiments, the presently disclosed subject matter provides pharmaceutical compositions comprising a compound of formula (I). In some embodiments, the pharmaceutical compositions comprise one or more pharma- ceutically acceptable carriers, diluents, excipients, or adjuvants.

[0084] In some embodiments, the subject matter of the present disclosure has formula (II): [ka] providing a compound of the formula: wherein each y is independently an integer selected from the group consisting of 0, 1, and 2; R 1x , R 2x , and R 3x’ , are H, OH, halogen, and C, respectively. 1-6 Alkyl, -OC 1-6 Alkyl, and -SC 1-6 independently selected from the group consisting of alkyl; R 4x is H, linear or branched C1 - 6 Alkyl, -(CH 2 ) q4 -aryl and hydroxyl substituted straight or branched chain C 1 - 6 alkyl, wherein q4 is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6; R 5x and R 6x are each independently H, a halogen, or -O-(CH 2 ) z3 -X a where z1 is an integer from 0 to 4, z2 is an integer from 0 to 2, z3 is an integer from 1 to 6, and X a is a halogen, where R 5x and R 6x cannot both be H at the same time; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0085] In some embodiments, the compound of Formula (II) has the following structure: [ka] Selected from.

[0086] In other embodiments, the subject matter of the present disclosure has formula (III): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is absent or is a targeting moiety for FAP-α, where A and B can be the same or different; C 1 may be absent or present, and when present is a chelating group; C 2 is a prosthetic group; L a and L b are each other as well as A, B, C 1 and C 2 is a bifunctional linker capable of forming a chemical bond with; L c1 and L c2 are independent of each other and A, B, L a , and L b is a bifunctional linker capable of forming a chemical bond with; So C 1 If absent, L c1 is also absent, and if B is absent then L b is also non-existent; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0087] In some embodiments, A is: [ka] or, if B is present, A and B each are; During the ceremony, [ka] is the linker L a and / or L b where the attachment point can be at any of carbon atoms 5, 6, 7, or up to 8 of the quinolinyl ring; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0088] In some embodiments, C 2 teeth 18 F, 124 I, 125 I, 131 I, and 211 A prosthetic group containing a radioisotope selected from the group consisting of At.

[0089] In some embodiments, the prosthetic group is: [ka] Selected from the group consisting of; where each X is a straight chain or a branched C 1 -C 8 Alkyl, -SO 2 , -C(=O)-, -C(=O)OR 20 are independently selected from the formula R 20 is H or C 1 -C 4 Alkyl, and 18 F, 124 I, 125 I, 131 I, and 211 At; each R and R' is defined above; and each n is independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, wherein [ka] Each carbon in the alkylene chain is C 1 -C 4 It can be substituted with alkyl.

[0090] In some embodiments, the prosthetic group is: [ka] is selected from the group consisting of:

[0091] In some embodiments, C 1These include DOTAGA (1,4,7,10-tetraazacyclododecane, 1-(glutaric acid)-4,7,10-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTASA (1,4,7,10-tetraazacyclododecane-1-(2-succinic acid)-4,7,10-triacetic acid), CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), DEPA (7-[2-(bis-carboxymethylamino)-ethyl]-4,10-bis-carboxymethyl-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), and CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane). 3p-C-DEPA (2-[(carboxymethyl)][5-(4-nitrophenyl-1-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]pentan-2-yl)amino]acetic acid)), TCMC (2-(4-isothiocyanotobenzyl)-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamonylmethyl)-cyclododecane), oxo-DO3A (1-oxa-4,7,10-triazacyclododecane-5-S-(4-isothiocyanatobenzyl)-4,7,10-triacetic acid), p-NH 2-Bn-oxo-DO3A (1-oxa-4,7,10-tetraazacyclododecane-5-S-(4-aminobenzyl)-4,7,10-triacetic acid), TE2A ((1,8-N,N′-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane), MM-TE2A, DM-TE2A, CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane), CB-TE1A1P (4,8,11-tetraazacyclotetradecane-1-(methyl)- CB-TE2P (1,4,8,11-tetraazacyclotetradecane-1,8-bis(methanephosphonic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N′,N″-triacetic acid), NODA (1,4,7-triazacyclononane-1,4-diacetate); NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), (NOTAGA) DFO (deferoxamine), 1,4,7-triazonane-1,4-diyl)diacetic acid, NETA ([4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonane-1-yl}-acetic acid), TACN-TM (N,N',N'',tris(2-mercaptoethyl)-1,4,7-triazacyclononane), Diamsar (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo(6,6,6)eicosane, 3,6,1 and a chelating agent selected from the group consisting of 0,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), Sarar (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), AmBaSar (4-((8-amino-3,6,10,13,16,19-hexazabicyclo[6.6.6]icosan-1-ylamino)methyl)benzoic acid), and BaBaSar.

[0092] In some embodiments, C 1 is a chelating agent and is: [ka] [ka] [ka] [ka] [ka] is a chelating agent is selected from the group consisting of.

[0093] In some embodiments, the chelating agent further comprises a radioactive metal.

[0094] In some embodiments, the radiometal 60 Cu, 62 Cu, 64 Cu, 67 Cu, 203 Pb, 212 Pb, 225 Ac, 177 Lu, 99m Tc, 68 Ga, 149 Tb, 86 Y, 90 Y, 111 In, 115 In, 186 Re, 188 Re, 153 Sm, 89 Zr, 213 Bi, 212 Bi, 212 Pb, 67 Ga, 47 Sc, 166 Ho, 43 Sc, 223 Ra, 226 / 227 Th, Al- 18 F, and Sc- 18 Selected from the group consisting of F.

[0095] In some embodiments, L a , L b , L c1 , and L c2 are each independently selected from the group consisting of (a), (b), (c), or (d): (a) [ka] and; During the ceremony: p 1 , p 2 , p 3 and p 4 may be in any order; t 1 and t 2 are each an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; p 1、 p 3 , and p 4 are each independently 0 or 1; p 2 is an integer selected from the group consisting of 0, 1, 2, and 3, and p 2 When is 2 or 3, each R 1 are the same or different; m 1 , m 2 , m 3 , and m 4 are each an integer independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; W 1 is selected from the group consisting of a bond, -S-, -C(=O)-, -C(=O)-NR-, and -NR-C(=O)-; W 2 is a bond, -S-, -CH 2 -C(=O)-NR-, -C(=O)-, -NRC(=O)-, -NR'C(=O)NR-, -NRC(=S)NR' 2 -, -NRC(=O)O-, -OC(=O)NR-, -OC(=O)-, -C(=O)NR-, -NR-C(=O)-, -C(=O)O-, -(O-CH 2 -CH 2 )q - and -(CH 2 -CH 2 -O) q wherein q is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; Each R or R' is independently H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and -OR 4 where R 4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl, where q is defined as immediately above; H can be present or absent, and when present, is selected from the group consisting of nitrogen-containing heteroalkylene chains, cyclic or bicylic heteroalkyl radicals, and the structures [ka] A triazole radical having the formula: Each R 1 Regardless of H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, -(CH 2 ) q -C 3 -C 12 Aryl, -C 4 -C 16 Alkylaryl, or -(CH 2 ) q -C 4 -C 16 R is alkylaryl; 2 and R 3 are independently H, -(CH 2 ) q -C 3 -C 12 Aryl, and -CO 2 R 5 where R 5 is H, C 1 -C6 Alkyl, C 3 -C 12 Aryl, and C 4 -C 16 alkylaryl, where R 2 and R 3 can be the same or different, where q is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; V is selected from the group consisting of -C(O)-, -C(S)-, -NRC(O)-, -NRC(S)-, and -OC(O)-; (b) [ka] and p in the formula 1、 p 2 , p 3 , m 1 , m 2 , Tz, W 2、 R, R 1 , R 2 , R 3 and V is defined above; (c)-L 1 -, -L 2 -L 3 - or -L 1 -L 2 -L 3 - in which: L 1 -NR-(CH 2 ) q -[O-CH 2 -CH 2 -O] q -(CH 2 ) q -C(=O)-; L 2 -NR-(CH 2 ) q -C(COOR 5 )-NR-; and L 3 is -(O=)C-(CH 2 ) q -C(=O)-; wherein each q is independently an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as defined above; and (d)-(CR 6 H) q -(CH 2 ) q -C(=O)-NR-(CH 2 ) q -O- or -NR-(CH 2 ) q -O-; where each q and R are defined above; and R 6 H or -COOR 5 It is.

[0096] In some embodiments, H is: [ka] where X 1 and X 2 are each independently -CH- or N; 16 is H or -C(=O)-OR regardless 17 where R 17 is C 1 -C 4 is alkyl; -N(R 18 )-(CH 2 ) z5 -N(R 19 )--wherein R 18 and R 19 are independently H or C 1 -C 4 alkyl, and Z5 is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; [ka] Selected from.

[0097] In some embodiments, one or more L a , L b , Lc1 , and L c2 The following: [ka] [ka] It includes one or more units selected from wherein u is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as set out above.

[0098] In some embodiments, the compound of Formula (III) is: [ka] [ka] [ka] Selected from.

[0099] In some embodiments, the prosthetic group C 2 is a chelating group C 1 covalently binds to

[0100] In some embodiments, the compound is: [ka] Selected from.

[0101] In some embodiments, the subject matter of the present disclosure has formula (IV): [ka] providing a compound of the formula: During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is a targeting moiety for prostate-specific membrane antigen (PSMA); C is any optically or radiolabeled functional group suitable for optical imaging, photoacoustic imaging, positron emission tomography (PET) imaging, single photon emission computed tomography (SPECT) imaging, or radiotherapy; and L a , L b , and L c are bifunctional linkers each capable of forming chemical bonds with each other and with A, B, and C, respectively; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0102] In some embodiments, A is: [ka] is a FAP-α targeting moiety having the structure: During the ceremony [ka] is the linker L a and / or L b where the attachment point can be at any of carbon atoms 5, 6, 7, or up to 8 of the quinolinyl ring; and stereoisomers and pharma- ceutically acceptable salts thereof.

[0103] In some embodiments, B has the structure: [ka] a targeting moiety for PSMA having the formula: During the ceremony: Z is tetrazole or CO 2 Q is; Q is H or a protecting group; q 1is an integer selected from the group consisting of 1, 2, 3, 4, and 5; q 2 is selected from an integer ranging from 0 to 1; and R y is independently H or -(CH 2 ) q3 -R y1 Where q 3 is an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, and 6, and R y1 is selected from the group consisting of substituted aryl, substituted pyridine, and unsubstituted isoquinoline;

[0104] In some embodiments, B has the structure: [ka] is a PSMA targeting moiety having the formula:

[0105] In some embodiments, R y1 is the following: [ka] Selected from the group consisting of; In the formula, X is Br, 75 Br, 76 Br, 77 Br, 80m Br, 82 Br, I, 124 I, 123 I, 125 I, 131 I, At, and 211 At.

[0106] In some embodiments, C is 18 F, 124 I, 125 I, 131 I, and 211 A radiolabeled prosthetic group containing a radioisotope selected from the group consisting of At.

[0107] In some embodiments, the radiolabeled prosthetic group is [ka] Selected from the group consisting of; In the formula, each X is independent 18 F, 124 I, 125 I, 131 I, and 211 At; each R and R' is defined above; and each n is independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0108] In some embodiments, the radiolabeled prosthetic group is: [ka] is selected from the group consisting of:

[0109] In some embodiments, C is selected from the group consisting of DOTAGA (1,4,7,10-tetraazacyclododecane, 1-(glutaric acid)-4,7,10-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTASA (1,4,7,10-tetraazacyclododecane-1-(2-succinic acid)-4,7,10-triacetic acid), CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), DEPA (7-[2-(bis-carboxymethylamino)-ethyl]-4,10-bis-carboxymethyl-1,4,7,10 -tetraaza-cyclododec-1-yl-acetic acid), 3p-C-DEPA (2-[(carboxymethyl)][5-(4-nitrophenyl-1-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl]pentan-2-yl)amino]acetic acid), TCMC (2-(4-isothiocyanotobenzyl)-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamonylmethyl)-cyclododecane), oxo-DO3A (1-oxa-4,7,10-triazacyclododecane-5-S-(4-isothiocyanatobenzyl)-4,7,10-triacetic acid), p-NH 2-Bn-oxo-DO3A (1-oxa-4,7,10-tetraazacyclododecane-5-S-(4-aminobenzyl)-4,7,10-triacetic acid), TE2A ((1,8-N,N′-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane), MM-TE2A, DM-TE2A, CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane), CB-TE1A1P (4,8,11-tetraazacyclotetradecane-1-(methyl)- CB-TE2P (1,4,8,11-tetraazacyclotetradecane-1,8-bis(methanephosphonic acid), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N′,N″-triacetic acid), NODA (1,4,7-triazacyclononane-1,4-diacetate); NODAGA (1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid), (NOTAGA) 1,4,7-Triazonan-1,4-diyl)diacetic acid DFO (deferoxamine), NETA ([4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid), TACN-TM (N,N',N'',tris(2-mercaptoethyl)-1,4,7-triazacyclononane), Diamsar (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo(6,6,6)eicosane, 3,6 ,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), Salal (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), Anbasar (4-((8-amino-3,6,10,13,16,19-hexazabicyclo[6.6.6]icosan-1-ylamino)methyl)benzoic acid), and BaBaSar.

[0110] In some embodiments, C is: [ka] [ka] [ka] [ka] [ka] The chelating agent is selected from the group consisting of:

[0111] In some embodiments, the chelating agent further comprises a radioactive metal.

[0112] In some embodiments, the radiometal 60 Cu, 62 Cu, 64 Cu, 67 Cu, 203 Pb, 212 Pb, 225 Ac, 177 Lu, 99m Tc, 68 Ga, 149 Tb, 86 Y, 90 Y, 111 In, 115 In, 186 Re, 188 Re, 153 Sm, 89 Zr, 213 Bi, 212 Bi, 212 Pb, 67 Ga, 47 Sc, 166 Ho, 43 Sc, 223 Ra, 226 / 227 Th, Al- 18 F, and Sc- 18 Selected from the group consisting of F.

[0113] In some embodiments, C includes an optical dye.

[0114] In some embodiments, the optical dye includes a fluorescent dye.

[0115] In some embodiments, the fluorescent dye comprises a fluorescent dye that emits in the near infrared region of the spectrum.

[0116] In some embodiments, the fluorescent dye is selected from the group consisting of polymethine dyes, coumarin dyes, xanthene dyes, and boron dipyrromethene (BODIPY) dyes.

[0117] In some embodiments, the polymethine dye is selected from the group consisting of carbocyanine dyes, indocarbocyanine dyes, oxacarbocyanine dyes, thiacarbocyanine dyes, and merocyanine dyes.

[0118] In some embodiments, the xanthene dye is selected from the group consisting of a fluorescein dye and a coumarin dye.

[0119] In some embodiments, the fluorescent dye is: BODIPY FL, BODIPY R6G, BODIPY TR, BODIPY TMR, BODIPY 493 / 503, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, and BODIPY 650 / 665; Cy3, Cy3.5, Cy5, Cy5.5, Cy7, and Cy7.5; VivoTag-645, VivoTag-680, VivoTag-S680, VivoTag-S750, VivoTag-800; Alexa Fluor 488, Alexa Fluor 532, Alexa Fluor 546, Alexa Fluor 555, Alexa Fluor 568, Alexa Fluor 594, Alexa Fluor 647, Alexa Fluor 660, Alexa Fluor 680, Alexa Fluor 700, Alexa Fluor 750, and Alexa Fluor790; Dy677, Dy676, Dy682, Dy752, Dy780; DyLight 350, DyLight 405, DyLight 488, DyLight 547, DyLight 550, DyLight 594, DyLight 633, DyLight 647, DyLight 650, DyLight 680, DyLight 755, and DyLight 800; HiLyte Fluor 405, HiLyte Fluor 488, HiLyte Fluor 532, HiLyte Fluor 555, HiLyte TM Fluor 594, HiLyte Fluor 647, HiLyte Fluor 680, HiLyte Fluor 750; IR800 (dimethyl{4-[1,5,5-tris(4-dimethylaminophenyl)-2,4-pentadienylidene]-2,5-cyclohexadien-1-ylidene}ammonium perchlorate), IRDye 650, IRDye 680RD, IRDye 680LT, IRDye 700, IRDye 700DX, IRDye 750, IRDye 800, IRDye 800CW, IRDye 800RS; and Selected from the group consisting of ADS1065A, ADS1075A, ADS775MI, ADS775MP, ADS775PI, ADS775PP, ADS780HO, ADS780WS, ADS785WS, ADS790WS, ADS795WS, ADS798SM, ADS800AT, ADS815EI, ADS830AT, ADS830WS, ADS832WS, ADS845MC, and ADS920MC.

[0120] In some embodiments, the optical dye is: [ka] [ka] is selected from the group consisting of:

[0121] In some embodiments, L a , L b , and L c are each independently selected from the group consisting of (a), (b), (c), or (d): (a) [ka] and; During the ceremony: p 1 , p 2 , p 3 and p 4 may be in any order; t 1 and t 2 are each an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; p 1、 p 3 , and p 4 are each independently 0 or 1; p 2 is an integer selected from the group consisting of 0, 1, 2, and 3, and p 2 When is 2 or 3, each R 1 are the same or different; m 1 , m 2 , m 3 , and m 4 are each an integer independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; W 1 is selected from the group consisting of a bond, -S-, -C(=O)-, -C(=O)-NR-, and -NR-C(=O)-; W 2 is a bond, -S-, -CH2 -C(=O)-NR-, -C(=O)-, -NRC(=O)-, -NR'C(=O)NR-, -NRC(=S)NR' 2 -, -NRC(=O)O-, -OC(=O)NR-, -OC(=O)-, -C(=O)NR-, -NR-C(=O)-, -C(=O)O-, -(O-CH 2 -CH 2 ) q - and -(CH 2 -CH 2 -O) q wherein q is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; Each R or R' is independently H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and -OR 4 where R 4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl, where q is defined as immediately above; H can be present or absent, and when present, is a nitrogen-containing heteroalkylene chain, a cyclic or bicyclic heteroalkyl radical, and the structure [ka] A triazole radical having the formula: Each R 1 Regardless of H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, -(CH 2 ) q -C 3 -C 12 Aryl, -C 4 -C 16 Alkylaryl, or -(CH 2 ) q -C 4 -C 16 R is alkylaryl;2 and R 3 are independently H, -(CH 2 ) q -C 3 -C 12 Aryl, and -CO 2 R 5 where R 5 is H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, and C 4 -C 16 alkylaryl, where R 2 and R 3 can be the same or different, where q is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; V is selected from the group consisting of -C(O)-, -C(S)-, -NRC(O)-, -NRC(S)-, and -OC(O)-; (b) [ka] and p in the formula 1、 p 2 , p 3 , m 1 , m 2 , Tz, W 2、 R, R 1 , R 2 , R 3 and V is defined above; (c)-L 1 -, -L 2 -L 3 - or -L 1 -L 2 -L 3 - in which: L 1 -NR-(CH 2 ) q -[O-CH 2 -CH 2 -O] q -(CH 2 ) q -C(=O)-; L2 -NR-(CH 2 ) q -C(COOR 5 )-NR-; and L 3 is -(O=)C-(CH 2 ) q -C(=O)-; wherein each q is independently an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as defined above; and (d)-(CR 6 H) q -(CH 2 ) q -C(=O)-NR-(CH 2 ) q -O- or -NR-(CH 2 ) q -O-; where each q and R are defined above; and R 6 H or -COOR 5 It is.

[0122] In some embodiments, H is: [ka] where X 1 and X 2 are each independently -CH- or N; 16 is H or -C(=O)-OR regardless 17 where R 17 is C 1 -C 4 is alkyl; -N(R 18 )-(CH 2 ) z5 -N(R 19 )--wherein R 18 and R 19 are independently H or C 1 -C 4 alkyl, and Z5 is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; [ka] Selected from.

[0123] In some embodiments, one or more L a , L b , and L c is the following: [ka] [ka] Contains one or more units selected from where u is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as set out above.

[0124] In some embodiments, the compound of Formula (IV) is: [ka] [ka] is selected from the group consisting of:

[0125] B.How to use B.1. Imaging Methods

[0126] In other embodiments, the subject matter of the present disclosure provides a method for imaging a disease or disorder associated with fibroblast activation protein-alpha (FAP-alpha) and / or prostate-specific membrane antigen (PSMA), the method comprising administering a compound of Formula (I-IV), or a pharmaceutical composition thereof, wherein the compound of Formula (I-IV) comprises an optically or radioactively labeled functional group suitable for optical imaging, photoacoustic imaging, PET imaging, or SPECT imaging; and obtaining an image.

[0127] Thus, in some embodiments, the subject matter of the present disclosure provides a method for imaging one or more cells, organs or tissues, the method comprises exposing cells to an effective amount of the compound of formula (I-IV) with suitable optical or radioisotope label for imaging or administering to a subject.In some embodiments, the one or more organs or tissues comprise prostate tissue, kidney tissue, brain tissue, vascular tissue or tumor tissue.

[0128] The imaging methods of the present invention are suitable for imaging any physiological process or feature in which FAP-α and / or PSMA is involved, for example, for identifying areas of tissue or targets that exhibit high concentrations or express FAP-α and / or PSMA.

[0129] Physiological processes in which FAP-α is involved include, but are not limited to: (a) proliferative diseases (including, but not limited to, cancer); (b) tissue remodeling and / or chronic inflammation (including, but not limited to, fibrotic diseases, wound healing, keloid formation, osteoarthritis, rheumatoid arthritis, and related disorders involving cartilage degradation); and (c) endocrinological disorders (including, but not limited to, disorders of glucose metabolism).

[0130] In certain embodiments, the radiolabeled compound is stable in vivo.

[0131] In certain embodiments, the radiolabeled compound is detected by positron emission tomography (PET) or single photon emission computed tomography (SPECT).

[0132] In certain embodiments, the optical reporting moiety is detected by fluorescence, such as, for example, by fluorescence microscopy.

[0133] In certain embodiments, the compounds of the present disclosure are rapidly excreted from the body tissues to prevent long-term exposure to the radiation of the radiolabeled compound administered to the subject.Typically, the compounds of the present disclosure are eliminated from the body in less than about 24 hours.More typically, the compounds of the present disclosure are eliminated from the body in less than about 16 hours, less than 12 hours, less than 8 hours, less than 6 hours, less than 4 hours, less than 2 hours, less than 90 minutes, or less than 60 minutes.Exemplary compounds are eliminated between about 60 minutes and about 120 minutes.In certain embodiments, the compounds of the present disclosure are stable in vivo such that substantially all, for example more than about 50%, 60%, 70%, 80%, or 90%, of the injected compound is not metabolized in the body before being excreted.

[0134] Additionally, for in vitro applications, such as in vitro diagnostic and research applications, body fluids and cell samples of the subject are preferably used, such as blood, urine or tissue samples from mammals, particularly primates, such as humans, or blood urine or tissue samples from animals listed for veterinary applications.

[0135] Other embodiments provide kits comprising a compound of formula (I-IV). In certain embodiments, the kit provides a packaged pharmaceutical composition comprising a pharma- ceutically acceptable carrier and a compound of formula (I-IV). In certain embodiments, the packaged pharmaceutical composition comprises the reactive precursors necessary to produce a compound of formula (I-IV) by combination with a radiolabeled precursor. Other packaged pharmaceutical compositions further comprise indicia comprising at least one of instructions for preparing a compound of formula (I-IV) from the provided precursors, instructions for using the composition to image cells or tissues expressing FAP-α or PSMA.

[0136] In certain embodiments, a kit is provided that includes about 1 to about 30 mCi of the radionuclide-labeled imaging agent described above in combination with a pharma- ceutically acceptable carrier. The imaging agent and carrier may be provided in solution or in lyophilized form. When the imaging agent and carrier of the kit are in lyophilized form, the kit may optionally include a sterile and physiologically acceptable reconstitution medium, such as water, saline, buffered saline, and the like. The kit may provide the compound of formula (I-IV) in solution or in lyophilized form, and these components of the kit may optionally include a stabilizer, such as NaCl, silicate, phosphate buffer, ascorbic acid, gentisic acid, and the like. In this embodiment, additional stabilization of the kit components may be provided, for example, by providing a reducing agent in an oxidation-resistant form. The determination and optimization of such stabilizers and stabilization methods are well within the level of skill in the art.

[0137] In certain embodiments, the kit provides a non-radiolabeled precursor that is combined with the radiolabeled reagent in situ.

[0138] The imaging agent may be used according to the method of the present disclosure by those skilled in the art. Images may be generated based on the difference in spatial distribution of the imaging agent that accumulates at the site when contacted with FAP-α and / or PSMA. The spatial distribution may be measured using any means suitable for the particular label, such as a gamma camera, a PET device, a SPECT device, and the like. The degree of accumulation of the imaging agent may be quantified using known methods for quantifying radioactive emissions or fluorescence. A particularly useful imaging approach employs more than one imaging agent to perform simultaneous studies.

[0139] In most cases, a detectably effective amount of the imaging agent of the present invention is administered to the subject. A "detectably effective amount" of an imaging agent is defined as an amount sufficient to obtain an acceptable image using equipment available for clinical use. A detectably effective amount of the imaging agent may be administered in more than one injection. A detectably effective amount of the imaging agent of the present invention may vary depending on factors such as the degree of sensitivity of the individual, the age, sex, and weight of the individual, the individual's idiosyncratic response, and dosimetry. A detectably effective amount of the imaging agent may also vary depending on factors related to the instrument and film. Optimization of such factors is well within the level of skill in the art. The amount of imaging agent used for diagnostic purposes and the duration of the imaging study will depend on the radionuclide used to label the imaging agent, the patient's weight, the nature and severity of the condition being treated, the nature of the therapeutic treatment the patient has undergone, and the patient's idiosyncratic response. Ultimately, the attending physician will determine the amount of imaging agent administered to each individual patient and the duration of the imaging study.

[0140] D. Methods of Treating Diseases or Disorders Associated with FAP-α and / or PSMA Using Compounds of Formula (I-IV), or Pharmaceutical Compositions Thereof

[0141] In some embodiments, the presently disclosed subject matter provides a method for inhibiting fibroblast activation protein-alpha (FAP-alpha) and / or prostate-specific membrane antigen (PSMA), the method comprising administering to a subject in need thereof an effective amount of a compound of Formula (I-IV), or a pharmaceutical composition thereof.

[0142] In other embodiments, the presently disclosed subject matter provides a method for treating a fibroblast activation protein-alpha (FAP-α)- and / or prostate-specific membrane antigen (PSMA)-associated disease or disorder, the method comprising administering to a subject in need of such treatment an effective amount of a compound of Formula (I-IV), or a pharmaceutical composition thereof, wherein the compound of Formula (I-IV) comprises a radiolabel functional group suitable for radiation therapy.

[0143] In some embodiments, the compounds of formula (I-IV) of the present disclosure can be used to treat a subject suffering from one or more FAP-α related diseases or disorders, including but not limited to the following: (a) proliferation (including but not limited to cancer); (b) tissue remodeling and / or chronic inflammation (including but not limited to fibrotic diseases, wound healing, keloid formation, osteoarthritis, rheumatoid arthritis and related disorders involving cartilage degeneration); and (c) endocrinological disorders (including but not limited to disorders of glucose metabolism).

[0144] Thus, in some embodiments, the one or more FAP-α related diseases or disorders are selected from the group consisting of proliferative diseases, including, but not limited to, breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, melanoma, fibrosarcoma, bone and connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, and adenocarcinoma; diseases characterized by tissue remodeling and / or chronic inflammation; diseases associated with endocrinological dysfunction; and blood clotting disorders.

[0145] In certain embodiments, the prostate-specific membrane antigen (PSMA)-related disease or disorder is selected from the group consisting of prostate cancer, renal cancer, head cancer, neck cancer, head and neck cancer, lung cancer, breast cancer, prostate cancer, colorectal cancer, esophageal cancer, gastric cancer, leukemia / lymphoma, uterine cancer, skin cancer, endocrine cancer, urinary cancer, pancreatic cancer, gastrointestinal cancer, ovarian cancer, cervical cancer, adenoma, and tumor neovasculature. In certain embodiments, the prostate-specific membrane antigen (PSMA)-related disease or disorder comprises prostate cancer.

[0146] In general, an "effective amount" of an active agent or drug delivery device refers to the amount necessary to elicit a desired biological response. As will be appreciated by those of ordinary skill in the art, the effective amount of an agent or device may vary depending on factors such as the desired biological endpoint, the agent being delivered, the composition of the agent composition, the target tissue, and the like.

[0147] The term "combination" is used in its broadest sense and means that a subject is administered at least two agents, more specifically, a compound described herein and at least one other therapeutic agent. More specifically, the term "in combination" refers, for example, to the concomitant administration of two (or more) active agents for the treatment of a single disease state. As used herein, the active agents may be administered in combination and in a single dosage form, simultaneously as separate dosage forms, or as separate dosage forms that are administered alternately or sequentially on the same or separate days. In one embodiment of the subject matter of the present disclosure, the active agents are administered in combination in a single dosage form. In another embodiment, the active agents are administered in separate dosage forms (for example, where it is desirable to vary the amount of one and not the other). The single dosage form may contain an additional active agent for the treatment of a disease state.

[0148] Additionally, the compounds described herein, alone or in combination with one or more therapeutic agents, can be administered in combination with adjuvants that enhance the stability of the compounds, in certain embodiments, facilitate administration of pharmaceutical compositions containing them, provide increased solubility or dispersibility, increase inhibitory activity, include other active ingredients, provide adjunctive therapy, and the like. Advantageously, such combination therapies utilize lower doses of conventional therapeutic substances, thus avoiding the toxicities and side effects that may be incurred when those agents are used as monotherapy.

[0149] The timing of administration of the compound described herein and at least one additional therapeutic agent can be varied, so long as the beneficial effects of the combination of these agents are achieved.Thus, the phrase "in combination with" refers to the administration of the compound described herein and at least one additional therapeutic agent simultaneously, sequentially, or any combination thereof.Thus, a subject who is administered a combination of the compound described herein and at least one additional therapeutic agent can receive the compound and at least one additional therapeutic agent at the same time (i.e., simultaneously) or at different times (i.e., sequentially, in any order, on the same day, or on different days), so long as the combined effects of both agents are achieved in the subject.

[0150] When administered sequentially, the agents can be administered within 1, 5, 10, 30, 60, 120, 180, 240 minutes or more of each other. In other embodiments, the agents administered sequentially can be administered within 1, 5, 10, 15, 20 or more days of each other. When the compounds described herein and at least one additional therapeutic agent are administered simultaneously, they can be administered to the subject as separate pharmaceutical compositions, each of which contains either the compound or the at least one additional therapeutic agent, or they can be administered to the subject as a single pharmaceutical composition containing both agents.

[0151] When administered in combination, the effective concentration of each agent to elicit a particular biological response may be lower than the effective concentration of each agent when administered alone, thereby reducing the dosage of one or more agents relative to the dosage required when the agent is administered as a single agent. The effect of multiple agents is not necessarily additive or synergistic. Agents may be administered multiple times.

[0152] In some embodiments, two or more agents can have a synergistic effect when administered in combination. As used herein, "synergy," "synergistic," "synergistically," and derivatives thereof refer to a situation in which the biological activity of a combination of a compound described herein and at least one additional therapeutic agent is greater than the sum of the biological activities of each agent when administered individually, e.g., in a "synergistic effect" or a "synergistic combination" or a "synergistic composition."

[0153] Synergy can be expressed in terms of a "Synergy Index (SI)", which is usually: Q a / Q A +Q b / Q B = Synergy Index (SI) The ratio can be calculated by the method described in F. C. Kull et al., Applied Microbiology 9, 538 (1961), During the ceremony: Q A is the concentration of component A acting alone that produces the end point relative to component A; Q a is the concentration of component A in the mixture that produces the endpoint; Q B is the concentration of component B acting alone that produces an end point relative to component B; and Q b is the concentration of component A in the mixture that produces the endpoint.

[0154] Usually, Q a / Q A and Q b / Q BWhen the sum is higher than one, it indicates antagonism. When the sum is equal to one, it indicates additivity. When the sum is less than one, it demonstrates synergy. The lower the SI, the greater the synergy exhibited by that particular mixture. Thus, a "synergistic combination" has greater activity than can be expected based on the activity of the individual components observed when used alone. Furthermore, a "synergistically effective amount" of a component refers to the amount of the component required to elicit a synergistic effect, for example, in another therapeutic agent present in the composition.

[0155] In other embodiments, the method can be practiced in vitro or ex vivo by introducing and preferably mixing the compound and the cell(s) or tumor(s) in a controlled environment such as a culture dish or tube. The method can be practiced in vivo, in which case contacting means exposing a target in a subject to at least one compound of the presently disclosed subject matter, such as administering the compound to the subject via any suitable route. According to the presently disclosed subject matter, contacting can consist of introducing, exposing, and the like, a compound at a site remote from the cells to be contacted, and allowing the subject's bodily functions, or natural (e.g., diffusion) or artificial (e.g., swirling) movement of fluids to bring the compound into contact with the target.

[0156] The subjects treated by the methods of the present disclosure are, in many of their embodiments, desirably human subjects, although it is understood that the methods described herein are effective with respect to all vertebrate species intended to be encompassed by the term "subject." Thus, a "subject" can include a human subject for medical purposes, such as, for example, treatment of an existing condition or disease or prophylactic treatment to prevent the onset of a condition or disease, or an animal (non-human) subject for medical, veterinary, or developmental purposes. Suitable animal subjects include, but are not limited to, primates, such as humans, monkeys, apes, and the like; bovines, such as cattle, oxen, and the like; ovines, such as sheep, and the like; caprines, such as goats, and the like; porcines, such as pigs, hogs, and the like; equines, such as horses, donkeys, zebras, and the like; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, and the like; and rodents, including mice, rats, and the like. The animals may be transgenic animals. In some embodiments, the subject is a human, including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Additionally, "subject" can include a patient suffering from or suspected of suffering from a condition or disease. Thus, the terms "subject" and "patient" are used interchangeably herein. In some embodiments, the subject is a human. In other embodiments, the subject is a non-human.

[0157] As used herein, the term "treating" can include reversing, alleviating, inhibiting the progression of, preventing, or reducing the likelihood of the disease, or condition to which such term applies, or one or more symptoms or manifestations of such disease or condition.

[0158] "Preventing" refers to making a disease, condition, or symptom or manifestation thereof, or worsening of such severity, not occur. Thus, the compounds of the present disclosure can be administered prophylactically to prevent or reduce the incidence or recurrence of a disease or condition.

[0159] C. Drug Composition and Administration

[0160] The present disclosure provides pharmaceutical compositions comprising one of the compounds of formula (I-IV) alone or in combination with one or more additional therapeutic agents in admixture with pharma-ceutically acceptable excipients. Those skilled in the art will recognize that the pharmaceutical compositions include pharma-ceutically acceptable salts of the above compounds. Pharmaceutically acceptable salts generally include salts of active compounds prepared using relatively non-toxic acids or bases that are well known to those of ordinary skill in the art and depending on the particular substituent moieties found on the compounds described herein. When the compounds of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a desired base, either neat or in a suitable inert solvent, or by ion exchange, whereby one basic counterion (base) in the ionic complex is replaced with another. Examples of pharma-ceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts.

[0161] When compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent, or by ion exchange, whereby one acidic counterion (the acid) in the ionic complex is replaced by another. Examples of pharma- ceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic, or phosphorous acid, and the like, as well as those derived from relatively non-toxic organic acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like. Also included are the salts of amino acids such as arginates and the like, and the salts of organic acids such as glucuronic or galactunoric acids and the like (see, e.g., Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0162] Thus, pharma- ceutically acceptable salts suitable for use with the subject matter of the present disclosure include, by way of example and without limitation, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, citrate, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinol, hexylsuccinate ... Other pharma- ceutically acceptable salts include, for example, phosphate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, or teoclate. ... th (Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000). In therapeutic and / or diagnostic applications, the compounds of the present disclosure can be formulated for a variety of modes of administration, including systemic administration and local or topical administration. Techniques and formulations are generally described in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000).

[0163] Depending on the particular condition to be treated, such drugs can be formulated into liquid or solid dosage forms and administered systemically or locally. Drugs can be delivered, for example, in time-release or sustained-release forms, as known to those skilled in the art. Techniques for formulation and administration are described in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000). Suitable routes include oral, buccal, inhalation spray, sublingual, rectal, transdermal, vaginal, transmucosal, nasal or intestinal administration; intramuscular, subcutaneous, intramedullary injection, and parenteral delivery, including intrathecal, direct intraventricular, intravenous, intra-articullar, intrasternal, intrasynovial, intrahepatic, intralesional, intracranial, intraperitoneal, intranasal or intraocular injection or other delivery modes.

[0164] For injection, the agents of the present disclosure can be formulated and diluted in aqueous solutions, such as, for example, Hank's solution, Ringer's solution, or physiologically compatible buffers such as physiological saline buffer, etc. For such transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.

[0165] The use of pharmaceutically acceptable inert carriers to formulate the disclosed compounds for the practice of the present disclosure into dosages suitable for systemic administration is within the scope of the present disclosure.By appropriate selection of carriers and appropriate manufacturing practices, the compositions of the present disclosure, particularly those formulated as solutions, can be administered parenterally, such as by intravenous injection.The present compounds can be easily formulated into dosages suitable for oral administration using pharmaceutically acceptable carriers well known in the art.Such carriers allow the compounds of the present disclosure to be formulated into tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, and the like, for oral ingestion by the subject (e.g., patient) to be treated.

[0166] For nasal or inhalation delivery, the agents of the present disclosure may also be formulated by methods known to those of skill in the art and may include, for example, but not limited to, solubilizing, diluting, or dispersing agents such as saline; preservatives such as benzyl alcohol; absorption enhancers; and fluorocarbons.

[0167] Suitable pharmaceutical compositions for use in the present disclosure include compositions in which the active ingredient is contained in an amount effective to achieve its intended purpose. Determination of an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. In general, compounds according to the present disclosure are effective over a wide dosage range. For example, in treating adult humans, dosages of 0.01 to 1000 mg per day, 0.5 to 100 mg, 1 to 50 mg, and 5 to 40 mg per day are examples of dosages that may be used. A non-limiting dosage is 10-30 mg per day. The exact dosage depends on the route of administration, the dosage form of the compound, the subject to be treated, the body weight of the subject to be treated, the bioavailability of the compound(s), the adsorption, distribution, metabolism, and excretion (ADME) toxicity of the compound(s), and the preference and experience of the attending physician.

[0168] In addition to the active ingredient, these pharmaceutical compositions can contain suitable pharma- ceutical acceptable carriers, including excipients and auxiliaries that facilitate the processing of the active compound into pharma- ceutical preparations. Preparations formulated for oral administration can be in the form of tablets, dragees, capsules, or solutions.

[0169] Pharmaceutical preparations for oral use can be obtained by combining active compounds with solid excipients, optionally grinding the resulting mixture, and then processing the mixture of granules to obtain tablets or dragee cores, after adding suitable auxiliaries as necessary.Suitable excipients are, in particular, sugars, such as lactose, sucrose, mannitol, or sorbitol; cellulose preparations, such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethyl-cellulose, sodium carboxymethyl-cellulose (CMC), and / or fillers such as polyvinylpyrrolidone (PVP: povidone).If necessary, disintegrants, such as cross-linked polyvinylpyrrolidone, agar, alginic acid, or its salts, such as sodium alginate, may be added.

[0170] The dragee cores are provided with a suitable coating. For this purpose, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol (PEG), and / or titanium dioxide, lacquer solutions, and concentrated sugar solutions, which may optionally contain suitable organic solvents or solvent mixtures, may be used. Dye-stuffs dyes or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of doses of active compounds.

[0171] Orally usable pharmaceutical preparations include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. Push-fit capsules can contain the active ingredient in admixture with fillers, such as lactose, binders, such as starch, and / or lubricants, such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compound can be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols (PEGs). Additionally, stabilizers may be added.

[0172] D. Definition

[0173] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the subject matter described herein belongs.

[0174] Although the following terms related to compounds of formula (I-IV) are believed to be well understood by those of ordinary skill in the art, the following definitions are provided to facilitate the description of the subject matter of the present disclosure. These definitions are intended to supplement and illustrate, rather than preclude, the definitions that would be apparent to those of ordinary skill in the art upon review of the present disclosure.

[0175] The term substituted, as used herein, whether preceded by the term "optionally" or not, and regardless of the substituent, refers to the ability to change one functional group on a molecule to another functional group, provided that the valences of all atoms are maintained, as recognized by one of skill in the art. When more than one position in any given structure can be substituted with more than one substituent selected from a particular group, the substituents can be the same or different at every position. Substituents can also be further substituted (e.g., a substituent on an aryl group can have another substituent off it, such as another aryl group that is further substituted at one or more positions).

[0176] Where substituents or linking groups are specified by conventional chemical formulas written from left to right, they are represented by, for example, -CH 2 O-OCH 2 -C(=O)O- is equivalent to -OC(=O)-; OC(=O)NR- is equivalent to -NRC(=O)O-, and the like.

[0177] When the term “independently selected” is used, the substituents being referred to (e.g., the group R 1 , R 2 , and the like, or variables such as "m" and "n" may be the same or different. For example, R 1 and R 2 can both be substituted alkyl, or R 1 is hydrogen, and R 2 can be substituted alkyl, and the like.

[0178] The terms "a", "an" or "a(n)" when used herein with respect to a group of substituents means at least one. For example, if a compound is substituted with "an" alkyl or aryl, then the compound is optionally substituted with at least one alkyl and / or at least one aryl. Furthermore, if a moiety is substituted with an R substituent, then the group may be referred to as "R-substituted." If a moiety is R-substituted, then the moiety is substituted with at least one R substituent, and each R substituent is optionally different.

[0179] A named "R" or group generally has the structure recognized in the art as corresponding to the group having that name, unless otherwise specified herein. For purposes of illustration, certain representative "R" groups, as described above, are defined below.

[0180] The description of the compounds of the present disclosure is limited by the principles of chemical bonding known to those skilled in the art.Therefore, when a group can be substituted with one or more of a plurality of substituents, such substitutions are selected to give compounds known to those skilled in the art as following the principles of chemical bonding and not being inherently unstable and / or not likely to be unstable under ambient conditions, such as aqueous, neutral, and some known physiological conditions.For example, heterocycloalkyl or heteroaryl is attached to the rest of the molecule through ring heteroatom according to the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0181] As used herein, unless expressly defined otherwise, a "substituent" includes a functional group selected from one or more of the following moieties, as defined herein:

[0182] The term hydrocarbon, as used herein, refers to any chemical group containing hydrogen and carbon. The hydrocarbon may be substituted or unsubstituted. Any substitutions must satisfy all valencies, as would be known to one of skill in the art. The hydrocarbon may be unsaturated, saturated, branched, unbranched, cyclic, polycyclic, or heterocyclic. Exemplary hydrocarbons are further defined herein below and include, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl, and the like.

[0183] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, straight-chain (i.e., unbranched) or branched-chain, acyclic or cyclic hydrocarbon groups, or combinations thereof, which may be fully saturated, mono- or polyunsaturated, and can include divalent and polyvalent groups, having the specified number of carbon atoms (i.e., C 1-10 In certain embodiments, the term "alkyl" refers to a group having from 1 to 10 carbons, including 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbons. 1-20 and 20 carbon atoms, inclusive, which may be linear (i.e., "straight chain"), branched, or cyclic, saturated or at least partially, and optionally fully unsaturated (i.e., alkenyl and alkynyl) derived by the removal of a single hydrogen atom from a hydrocarbon moiety containing between 1 and 20 carbon atoms.

[0184] Representative saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof.

[0185] "Branched" refers to an alkyl group having a lower alkyl group, such as methyl, ethyl, or propyl, attached to a linear alkyl chain. A "lower alkyl" refers to an alkyl group having from 1 to about 8 carbon atoms (i.e., C 1-8 "Alkyl" refers to alkyl groups having from about 10 to about 20 carbon atoms, examples being 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. In certain embodiments, "alkyl" refers specifically to alkyl groups having from about 10 to about 20 carbon atoms, examples being 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. 1-8 In other embodiments, "alkyl" refers specifically to a straight chain alkyl. 1-8 Reference is made to branched chain alkyls.

[0186] An alkyl group can be optionally substituted with one or more alkyl group substituents, which can be the same or different ("substituted alkyl"). The term "alkyl group substituted" includes, but is not limited to, alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms can be optionally inserted along the alkyl chain, where the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as "alkylaminoalkyl"), or aryl.

[0187] Thus, as used herein, the term "substituted alkyl," as defined herein, includes alkyl groups in which one or more atoms or functional groups of the alkyl group have been replaced with another atom or functional group, including, for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, cyano, and mercapto.

[0188] The term "heteroalkyl", by itself or in combination with another term, means, unless otherwise stated, a stable linear or branched chain or cyclic hydrocarbon radical having from 1 to 20 carbon atoms or heteroatoms, or a combination thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si and S, and in which the nitrogen, phosphorus and sulfur atoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized. The heteroatom(s) O, N, P and S and Si may be located at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Examples include, but are not limited to, -CH 2 -CH 2 -O-CH 3 , -CH 2 -CH 2 -NH-CH 3 , -CH 2 -CH 2 -N(CH 3 )-CH 3 , -CH 2 -S-CH 2 -CH 3 , -CH 2 -CH 2 -S(O)-CH 3 , -CH 2 -CH 2 -S(O) 2 -CH 3 , -CH=CH-O-CH 3 , -Si(CH 3 ) 3 , -CH 2 -CH=N-OCH 3, -CH=CH-N(CH 3 )-CH 3 , O-CH 3 , -O-CH 2 -CH 3 Up to two or three heteroatoms are included, for example, -CH 2 -NH-OCH 3 and -CH 2 -O-Si(CH 3 ) 3 It may be continuous, such as:

[0189] As noted above, heteroalkyl groups, as used herein, can include heteroatoms, such as, for example, -C(O)NR', ​​-NR'R'', -OR', -SR, -S(O)R, and / or -S(O 2 )R', and the like. Where "heteroalkyl" is recited followed by a recitation of a specific heteroalkyl group, such as NR'R, or the like, it will be understood that the terms heteroalkyl and NR'R'' are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is recited for added clarity. Thus, the term "heteroalkyl" should not be construed herein as excluding specific heteroalkyl groups, such as NR'R'', and the like.

[0190] "Cyclic" and "cycloalkyl" refer to a non-aromatic monocyclic or polycyclic ring system of from about 3 to about 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Cycloalkyl groups can be optionally partially unsaturated. Cycloalkyl groups can also be optionally substituted with alkyl group substituents, oxo, and / or alkylene, as defined herein. One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms can be optionally inserted along the cyclic alkyl chain, where the nitrogen substituents are hydrogen, unsubstituted alkyl, substituted alkyl, aryl, or substituted aryl, thus providing a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Polycyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl, as well as fused ring systems, e.g., dihydro- and tetrahydronaphthalene, and the like.

[0191] The term "cycloalkylalkyl" as used herein refers to a cycloalkyl group, as defined above, which includes an alkylene moiety, also as defined above, examples of which include C 1-20 It is attached to the parent molecular moiety through an alkylene moiety. Examples of cycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl.

[0192] The term "cycloheteroalkyl" or "heterocycloalkyl" refers to a non-aromatic ring system, unsaturated or partially unsaturated ring system, e.g., a substituted or unsubstituted cycloalkyl ring system, from 3 to 10 members, containing one or more heteroatoms, which can be the same or different, and are selected from the group consisting of nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), and silicon (Si), and can optionally contain one or more double bonds.

[0193] Cycloheteroalkyl rings can be optionally fused or otherwise attached to other cycloheteroalkyl rings and / or non-aromatic hydrocarbon rings. Heterocyclic rings include those having from 1 to 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, where the nitrogen and sulfur heteroatoms can be optionally oxidized and the nitrogen heteroatom can be optionally quaternized. In certain embodiments, the term heterocyclic refers to a non-aromatic 5-, 6-, or 7-membered ring or polycyclic group in which at least one ring atom is a heteroatom selected from O, S, and N, wherein the nitrogen and sulfur heteroatoms may be optionally oxidized, including, but not limited to, fused 6-membered rings containing bi- or tri-cyclic groups and having between 1 and 3 heteroatoms independently selected from oxygen, sulfur, and nitrogen, where (i) each 5-membered ring has zero to two double bonds, each 6-membered ring has zero to two double bonds, and each 7-membered ring has zero to three double bonds, (ii) the nitrogen and sulfur heteroatoms may be optionally oxidized, (iii) the nitrogen heteroatom may be optionally quaternized, and (iv) any of the above heterocycles may be fused to an aryl or heteroaryl ring. Representative cycloheteroalkyl ring systems include, but are not limited to, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, indolinyl, quinuclidinyl, morpholinyl, thiomorpholinyl, thiadiazinanyl, tetrahydrofuranyl, and the like.

[0194] The terms "cycloalkyl" and "heterocycloalkyl", by themselves or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl", respectively. 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, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. The terms "cycloalkylene" and "heterocycloalkylene" refer to the divalent derivatives of cycloalkyl and heterocycloalkyl, respectively.

[0195] Unsaturated hydrocarbons have one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. Alkyl groups that are limited to hydrocarbon groups are referred to as "homoalkyl".

[0196] More specifically, the term "alkenyl" as used herein refers to a C alkyl group having at least one carbon-carbon double bond obtained by the removal of a hydrogen molecule. 2-20It refers to a monovalent group derived from an inclusive straight or branched hydrocarbon moiety. Alkenyl groups include, for example, ethenyl (i.e., vinyl), propenyl, butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, allenyl, and butadienyl.

[0197] The term "cycloalkenyl" as used herein refers to a cyclic hydrocarbon containing at least one carbon-carbon double bond. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.

[0198] The term "alkynyl" as used herein refers to a straight or branched alkyl group having a specified number of carbon atoms containing at least one carbon-carbon triple bond. 2-20 Refers to a univalent group derived from a hydrocarbon. Examples of "alkynyl" include ethynyl, 2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl, and heptynyl groups, and the like.

[0199] The term "alkylene" by itself or as part of another substituent refers to a straight or branched divalent aliphatic hydrocarbon group derived from an alkyl group having from 1 to about 20 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. The alkylene group can be straight, branched, or cyclic. The alkylene group can also be optionally unsaturated and / or substituted with one or more "alkyl group substitutions." One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms (also referred to herein as "alkylaminoalkyl") can be optionally inserted along the alkylene group, where the nitrogen substituent is alkyl as described above. Exemplary alkylene groups include methylene (-CH 2 -);Ethylene (-CH 2-CH 2 -); Propylene (-(CH 2 ) 3 -);Cyclohexylene (-C 6 H 10 -);-CH=CH-CH=CH-;-CH=CH-CH 2 -;-CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH=CHCH 2 -, -CH 2 CsCCH 2 -, -CH 2 CH 2 CH(CH 2 CH 2 CH 3 )CH 2 -, -(CH 2 ) q -N(R)-(CH 2 ) r -, where each of q and r is independently an integer from 0 to about 20, examples being 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, and R is hydrogen or lower alkyl; methylenedioxyl (-O-CH 2 -O-); and ethylenedioxyl (-O-(CH 2 ) 2 -O-). Alkylene groups can have from about 2 to about 3 carbon atoms, and can further have from 6 to 20 carbon atoms. Typically, alkyl (or alkylene) groups have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being some embodiments of the present disclosure. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having eight or fewer carbon atoms.

[0200] The term "heteroalkylene" by itself or as part of another substituent refers to heteroalkyl, such as, but not limited to, -CH 2 -CH 2 -S-CH 2-CH 2 - and -CH 2 -S-CH 2 -CH 2 -NH-CH 2 - means a divalent group derived from. In the case of heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (examples include alkyleneoxo, alkylenedioxo, alkyleneamino, alkylenediamino, and the like). Still further, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-.

[0201] The term "aryl", unless otherwise stated, means an aromatic hydrocarbon substituent, which may be a single ring or multiple rings (such as, for example, 1 to 3 rings, etc.), which are fused or covalently linked together. The term "heteroaryl" refers to an aryl group (or ring) containing from 1 to 4 heteroatoms (in each separate ring in the case of multiple rings) selected from N, O, and S, in which the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. Heteroaryl groups can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, Examples include 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" refer to the divalent forms of aryl and heteroaryl, respectively.

[0202] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the terms "arylalkyl" and "heteroarylalkyl" include aryl or heteroaryl groups in which the alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, furylmethyl, and the like) and those alkyl groups in which a carbon atom (e.g., methylene) is replaced with, for example, an oxygen atom (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, and the like). However, the term "haloaryl" as used herein is meant to cover only aryl substituted with one or more halogens.

[0203] When a heteroalkyl, heterocycloalkyl, or heteroaryl contains a specific number of members (eg, from 3 to 7 members), the term "member" refers to a carbon or heteroatom.

[0204] Further, generally, the formula: [ka] The structure represented by refers to ring structures, such as, but not limited to, aliphatic and / or aromatic ring structures, including 3-carbon, 4-carbon, 5-carbon, 6-carbon, 7-carbon, and the like, but including saturated, partially saturated, and unsaturated ring structures, including substituent R groups, where R groups may be present or absent, and when present, one or more R groups may each be substituted on one or more available carbon atoms of the ring structure. The presence or absence of R groups and the number of R groups is determined by the value of the variable "n", which is generally an integer having a value ranging from 0 to the number of carbon atoms on the ring available for substitution. Each R group, when more than one, is substituted on an available carbon of the ring structure rather than another R group. For example, the above structures, where n is from 0 to 2, include, but are not limited to, the following: [ka] and other similar compounds.

[0205] A dashed line representing a bond in a cyclic ring structure indicates that the bond is either present or absent in the ring, i.e., a dashed line representing a bond in a cyclic ring structure indicates that the ring structure is selected from the group consisting of saturated ring structures, partially saturated ring structures, and unsaturated ring structures.

[0206] symbol [ka] denotes the point of attachment of the moiety to the rest of the molecule.

[0207] When a named atom of an aromatic or heteroaromatic ring is defined as "absent," the named atom is replaced by a direct bond.

[0208] Each of the above terms (e.g., "alkyl," "heteroalkyl," "cycloalkyl," and "heterocycloalkyl," "aryl," "heteroaryl," "phosphonate," and "sulfonate" and their divalent derivatives) are meant to include both substituted and unsubstituted forms of the indicated radical. Optional substituents for each type of radical are provided below.

[0209] Substituents for alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl monovalent and divalent derivative groups (including those groups often referred to as alkylene, alkenyl, heteroalkylene, heteroalkenyl, alkynyl, cycloalkyl, heterocycloalkyl, cycloalkenyl, and heterocycloalkenyl) include, but are not limited to, the following: -OR', ═O, ═NR', ═N-OR', -NR'R'', -SR', -halogen, -SiR'R''R''', -OC(O)R', -C(O)R', -CO 2R', -C(O)NR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR''C(O)OR'-NR-C(NR'R'')=NR''', -S(O)R', -S(O) 2 R', -S(O) 2 NR'R'', -NRSO 2 R', -CN, CF 3 , Fluorinated C 1-4 Alkyl, and -NO, a number ranging from zero to (2m'+l). 2 where m' is the total number of carbon atoms in such group. R', R'', R''', and R'''' may each independently refer to hydrogen, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl (e.g., aryl substituted with 1-3 halogens), substituted or unsubstituted alkyl, alkoxy or thioalkoxy groups, or arylalkyl groups. As used herein, an "alkoxy" group is an alkyl attached to the remainder of the molecule through a divalent oxygen. When compounds of the present disclosure contain more than one R group, for example, each R group is independently selected, as are each R', R'', R''', and R'''' groups when more than one of these groups is present. When R' and R'' are attached to the same nitrogen atom, they can be combined with the nitrogen atom to form a 4-, 5-, 6-, or 7-membered ring. For example, -NR'R'' is meant to include, but not be limited to, 1-pyrrolidinyl and 4-morpholinyl. From the discussion of substituents above, those skilled in the art will appreciate that the term "alkyl" can also include haloalkyl (e.g., -CF 3 and -CH 2 CF 3 ) and acyls (e.g., -C(O)CH 3 , -C(O)CF 3 , -C(O)CH 2 OCH 3 It will be understood that the term "aryl" refers to groups that contain carbon atoms bonded to groups other than hydrogen groups, such as aryl, aryloxy ...

[0210] Similar to the substituents described for the alkyl groups above, exemplary substituents for the aryl and heteroaryl groups (and divalent derivatives thereof) are varied and include, for example: halogen, -OR', -NR'R'', -SR', SiR'R''R''', -OC(O)R', -C(O)R', -CO 2 R', -C(O)NR'R'', -OC(O)NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR''C(O)OR', -NR-C(NR'R''R''')=NR'''', NRC(NR'R'')=NR'''-S(O)R', -S(O) 2 R', -S(O) 2 NR'R'', -NRSO 2 R', -CN and -NO 2 , R', -N 3 , -CH(Ph) 2 , Fluoro(C 1-4 ) alkoxo, and fluoro (C 1-4 ) alkyl, selected in a number ranging from 0 to the total number of open valences on the aromatic ring system; and where R', R'', R''', and R'''' can be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl. When a compound of the present disclosure includes more than one R group, for example, each R group is independently selected, as are each R', R'', R''', and R'''' groups when more than one of these groups is present.

[0211] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring are optionally represented by the formula -TC(O)-(CRR') q -U-, where T and U are independently -NR-, -O-, -CRR'- or a single bond, and q is an integer from 0 to 3. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally form a ring of the formula -A-(CH 2 )r A and B are independently substituted by the substituents -CRR'-, -O-, -NR-, -S-, -S(O)-, -S(O) 2 -, -S(O) 2 NR'- or a single bond, and r is an integer from 1 to 4.

[0212] One of the single bonds of the new ring so formed may optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a group of the formula -(CRR') s X'-(CR') d -, where s and d are independently integers from 0 to 3, and X' is -O-, -NR'-, -S-, -S(O)-, -S(O) 2 - or -S(O) 2 The substituents R, R', R'', and R''' may be independently selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted heteroaryl.

[0213] As used herein, the term "acyl" refers to an organic acid group in which the -OH of the carboxyl group is replaced with another substituent and has the general formula RC(=O)-, where R is an alkyl, alkenyl, alkynyl, aryl, carbocylic, heterocyclic, or aromatic heterocyclic group as defined herein. As such, the term "acyl" specifically includes arylacyl groups such as 2-(furan-2-yl)acetyl- and 2-phenylacetyl groups. Specific examples of acyl groups include acetyl and benzoyl. Acyl groups are also intended to include amides, -RC(=O)NR', ​​esters, -RC(=O)OR', ketones, -RC(=O)R', and aldehydes, -RC(=O)H.

[0214] The terms "alkoxyl" or "alkoxy" are used interchangeably herein and refer to a saturated (i.e., alkyl-O-) or unsaturated (i.e., alkenyl-O- and alkynyl-O-) group attached to a parent molecular moiety through an oxygen atom, where the terms "alkyl", "alkenyl", and "alkynyl" are as previously described and include C 1-20 It can include straight, branched, or cyclic, saturated or unsaturated oxo-hydrocarbon chains, including, for example, methoxyl, ethoxyl, propoxyl, isopropoxyl, n-butoxyl, sec-butoxyl, tert-butoxyl, and n-pentoxyl, neopentoxyl, n-hexoxyl, and the like.

[0215] The term "alkoxyalkyl," as used herein, refers to an alkyl-O-alkyl ether, for example, a methoxyethyl or an ethoxymethyl group.

[0216] "Aryloxyl" refers to an aryl-O group as described above, including substituted aryl. As used herein, the term "aryloxyl" can refer to phenyloxyl or hexyloxyl, as well as phenyloxyl or hexyloxyl substituted with alkyl, substituted alkyl, halo, or alkoxyl.

[0217] "Aralkyl" refers to an aryl-alkyl group, where aryl and alkyl are as previously described, and includes substituted aryl and substituted alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0218] "Aralkyloxyl" refers to an aralkyl-O- group, in which the aralkyl group is as previously described. An exemplary aralkyloxyl group is benzyloxyl, i.e., C 6 H 5 CH 2 The aralkyloxyl group is optionally substituted.

[0219] "Alkoxycarbonyl" refers to the alkyl-OC(=O)- group. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butyloxycarbonyl, and tert-butyloxycarbonyl.

[0220] "Aryloxycarbonyl" refers to the group aryl-OC(=O)-. Exemplary aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.

[0221] "Aralkoxycarbonyl" refers to the group aralkyl-OC(=O)-. An exemplary aralkoxycarbonyl group is benzyloxycarbonyl.

[0222] "Carbamoyl" is a group of the formula -C(=O)NH 2 "Alkylcarbamoyl" refers to the group R'RN-C(=O)- where one of R and R' is hydrogen and the other R and R' is alkyl and / or substituted alkyl as described above. "Dialkylcarbamoyl" refers to the group R'RN-C(=O)- where each R and R' is independently alkyl and / or substituted alkyl as described above.

[0223] The term carbonyldioxyl, as used herein, refers to a carbonate group of formula -OC(=O)-OR.

[0224] "Acyloxyl" refers to an acyl-O- group, where acyl is as previously described.

[0225] The term "amino" means -NH 2 The term "amino" refers to a group, and also refers to nitrogen-containing groups as are known in the art, derived from ammonia by replacement of one or more hydrogen radicals with an organic radical. For example, the terms "acylamino" and "alkylamino" refer to certain N-substituted organic radicals with acyl and alkyl substituents, respectively.

[0226] The term "aminoalkyl" as used herein refers to an amino group covalently bonded to an alkylene linker. More specifically, the terms alkylamino, dialkylamino, and trialkylamino as used herein refer to one, two, or three, respectively, alkyl groups, as defined above, attached to a parent molecular moiety through a nitrogen atom. The term alkylamino refers to a group having the structure -NHR', where R' is an alkyl group, as defined above; while the term dialkylamino refers to a group having the structure -NR'R'' where R' and R'' are each independently selected from the group consisting of alkyl groups. The term trialkylamino refers to a group having the structure -NR'R''R'' where R', R'', and R''' are each independently selected from the group consisting of alkyl groups. Additionally, R', R'', and / or R'' taken together are optionally -(CH 2 ) k -, where k is an integer from 2 to 6. Examples include, but are not limited to, methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino, and propylamino.

[0227] An amino group is -NR'R'', where R' and R'' are typically selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0228] The terms alkylthioether and thioalkoxyl refer to saturated (i.e., alkyl-S-) or unsaturated (i.e., alkenyl-S- and alkynyl-S-) groups attached to a parent molecular moiety through a sulfur atom. Examples of thioalkoxyl moieties include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

[0229] "Acylamino" refers to an acyl-NH- group where acyl is as previously described. "Aroylamino" refers to an aroyl-NH- group where aroyl is as previously described.

[0230] The term "carbonyl" refers to a -C(=O)- group and can include aldehyde groups represented by the general formula RC(=O)H.

[0231] The term "carboxyl" refers to a -COOH group. Such groups are also referred to herein as "carboxylic acid" moieties.

[0232] The term "cyano" refers to the group -C≡N.

[0233] The terms "halo", "halide" or "halogen" as used herein refer to fluoro, chloro, bromo, and iodo groups. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C 1-4 )Alkyl" is defined to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0234] The term "hydroxyl" refers to an --OH group.

[0235] The term "hydroxyalkyl" refers to an alkyl group substituted with an --OH group.

[0236] The term "mercapto" refers to the group --SH.

[0237] The term "oxo" as used herein means an oxygen atom that is double bonded to a carbon atom or to another element.

[0238] The term "nitro" means -NO 2 Refer to the base.

[0239] The term "thio" refers to a compound described herein above in which a carbon or oxygen atom is replaced by a sulfur atom.

[0240] The term "sulfate" is -SO 4 Refer to the base.

[0241] The term thiohydroxyl or thiol, as used herein, refers to a group of formula --SH.

[0242] More specifically, the term "sulfide" refers to a compound having a group of formula -SR.

[0243] The term "sulfone" refers to the sulfonyl group -S(O 2 ) R

[0244] The term "sulfoxide" refers to a compound that has a sulfinyl group, -S(O)R.

[0245] The term ureido has the formula -NH-CO-NH 2 This refers to the urea group.

[0246] Throughout this specification and the claims, a given chemical formula or name is intended to encompass all tautomers, congeners, and optical- and stereoisomers, as well as racemic mixtures where such isomers and mixtures exist.

[0247] Certain compounds of the present disclosure may have asymmetric carbon atoms (optical or asymmetric centers) or double bonds, and enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomers that may be defined in terms of absolute stereochemistry as (R)- or (S)-, or for amino acids as D- or L-, and individual isomers are encompassed within the scope of the present disclosure. Compounds of the present disclosure do not include those known in the art to be too unstable to synthesis and / or separation. The present disclosure is intended to include racemic, scalemic, and optically pure forms of the compounds. Optically active (R)- and (S)-, or D- and L-isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques. When compounds described herein contain olefinic bonds or other centers of geometric asymmetry, and unless otherwise specified, the compounds are intended to include both E and Z geometric isomers.

[0248] Unless otherwise stated, structures depicted herein are intended to include all stereochemical forms of the structure, i.e., the R and S configurations for each asymmetric center. Thus, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0249] It will be apparent to one of skill in the art that certain compounds of the present disclosure may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present disclosure. The term "tautomer," as used herein, refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0250] Unless otherwise stated, structures depicted herein are meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. For example, replacement of hydrogen with deuterium or tritium, or replacement of carbon with 13 C- or I4Compounds having this structure substituted with C-enriched carbons are within the scope of this disclosure.

[0251] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain tritium ( 3 H), Iodine-125( 125 I) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.

[0252] The compounds of the present disclosure may exist as salts. The present disclosure includes such salts. Examples of applicable salt forms include hydrochloride, hydrobromide, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate (e.g., (+)-tartrate, (-)-tartrate, or mixtures thereof, including racemic mixtures), succinate, benzoate, and salts with amino acids such as glutamic acid. These salts can be prepared by methods known to those skilled in the art. Also included are base addition salts such as sodium salt, potassium salt, calcium salt, ammonium salt, organic amino salt, or magnesium salt, or similar salts. When the compounds of the present disclosure contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either neat or in a suitable inert solvent, or by ion exchange. Examples of acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic, or phosphorous acids and the like, as well as those derived from organic acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Also included are salts of amino acids such as arginates and the like, and salts of organic acids such as glucuronic or galactunoric acids and the like. Certain specific compounds of the present disclosure contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0253] The neutral forms of the compounds may 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.

[0254] Certain compounds of the present disclosure can exist in solvated forms, including hydrated forms, as well as nonsolvated forms.In most cases, solvated forms are equivalent to nonsolvated forms and are included within the scope of the present disclosure.Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the use contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0255] In addition to salt forms, the present disclosure provides compounds in the form of prodrugs. Prodrugs of the compounds described herein are compounds that undergo chemical changes under physiological conditions to provide the compounds of the present disclosure. Additionally, prodrugs can be converted to the compounds of the present disclosure by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to the compounds of the present disclosure when placed in a transdermal patch reservoir with the appropriate enzyme or chemical reagent.

[0256] The term "protecting group" refers to a chemical moiety that blocks some or all reactive moieties of a compound and prevents such moieties from participating in a chemical reaction until the protecting group is removed, for example, those moieties listed and described in TW Greene, PGM Wuts, Protective Groups in Organic Synthesis, 3rd ed. John Wiley & Sons (1999). When different protecting groups are employed, it can be advantageous for each (different) protecting group to be removable by different means. Protecting groups that are cleaved under entirely disparate reaction conditions allow for differential removal of such protecting groups. For example, protecting groups can be removed by acid, base, hydrogenolysis. Groups such as trityl, dimethoxytrityl, acetal, and tert-butyldimethylsilyl are acid labile and may be used to protect carboxy and hydroxy reactive moieties in the presence of hydrogenolysis removable Cbz groups and amino groups protected with base labile Fmoc groups. Carboxylic acid and hydroxy reactive moieties may be blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl in the presence of amines blocked with acid labile groups such as tert-butyl carbamate, or with acid and base stable but hydrolytically removable carbamates.

[0257] Carboxylic acid and hydroxy reactive moieties may also be blocked with hydrolytically removable protecting groups such as benzyl groups, while amine groups capable of hydrogen bonding with acids may be blocked with base labile groups such as Fmoc. Carboxylic acid reactive moieties may be blocked with oxidatively removable protecting groups such as 2,4-dimethoxybenzyl, and coexisting amino groups may be blocked with fluorine labile silyl carbamates.

[0258] Allyl blocking groups are useful in the presence of acid- and base-protecting groups because they are stable and can be subsequently removed by metal or pi-acid catalysts. For example, allyl-blocked carboxylic acids can be deprotected by palladium(O) catalysis in the presence of acid-labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate may be attached. As long as the residue is attached to the resin, that functional group is blocked and cannot react. Once released from the resin, the functional group is available for reaction.

[0259] Exemplary blocking / protecting groups include, but are not limited to, the following moieties: [ka]

[0260] Following long-standing patent law convention, the terms "a," "an," and "the" refer to "one or more" when used in this application, including the claims. Thus, for example, a reference to "an object" includes a plurality of objects unless the context clearly indicates the contrary (e.g., a plurality of objects), and so forth.

[0261] Throughout this specification and the claims, the terms "comprise," "comprises," and "comprising" are used in a non-exclusive sense, unless the context otherwise requires. Similarly, the term "include" and its grammatical variations are intended to be open-ended, and the recitation of items in a list does not exclude other similar items that may be substituted for or added to the listed items.

[0262] For purposes of this specification and the appended claims, unless otherwise indicated, all numbers expressing amounts, sizes, dimensions, ratios, shapes, formulations, parameters, percentages, quantities, properties, and other numerical values ​​used in this specification and claims are understood to be modified in all instances by the term "about," even if the term "about" is not explicitly stated in conjunction with the value, amount, or range. Thus, unless indicated to the contrary, the numerical parameters set forth in the following specification and the appended claims are not, and need not be, precise, but may be approximate and / or larger or smaller as desired, reflecting tolerances, conversion factors, rounding, measurement error and the like, as well as other factors known to those of skill in the art, depending upon the desired properties sought to be obtained by the subject matter of the present disclosure. For example, the term "about" when referring to a value can be defined to encompass variations such as, in some embodiments, ±100%, in some embodiments, ±50%, in some embodiments, ±20%, in some embodiments, ±10%, in some embodiments, ±5%, in some embodiments, ±1%, in some embodiments, ±0.5%, and in some embodiments, ±0.1% from the stated amount, as variations are appropriate for performing the disclosed methods or employing the disclosed compositions.

[0263] Additionally, when used in connection with one or more numbers or numerical ranges, the term "about" should be understood to refer to all such numbers, including all numbers within the range, and to modify the range by extending the boundaries above and below the recited numerical values. Recitation of numerical ranges by endpoints includes all numbers included within that range, examples and fractions thereof, integers (e.g., recitation of 1 through 5 includes 1, 2, 3, 4, and 5, as well as fractions thereof, examples being 1.5, 2.25, 3.75, 4.1, and the like), and any ranges within that range. EXAMPLES

[0264] The following examples are included to provide guidance to those of ordinary skill in the art to practice representative embodiments of the subject matter of the present disclosure. In light of this disclosure and the general level of skill in the art, the skilled artisan can recognize that the following examples are intended to be illustrative only, and that numerous variations, modifications, and alterations can be adopted without departing from the scope of the subject matter of the present disclosure. The synthetic descriptions and specific examples that follow are intended for illustrative purposes only, and are not to be construed as limiting in any way the making of the compounds of the present disclosure by other methods.

[0265] Example 1 Heterovalent FAP and PSMA Ligands 1.1 Experimental procedure and spectral characterization

[0266] Methyl (6-hydroxyquinoline-4-carbonyl)glycinate (2): 6-hydroxyquinoline-4-carboxylic acid (1) (200 mg, 1.05 mmol, 1.0 eq.), glycine methyl HCl salt (200 mg, 1.58 mmol, 1.5 eq.) and HATU (603 mg, 1.58 mmol, 1.5 eq.) were dissolved in 5 mL anhydrous DMF. To the solution was added DIPEA (0.46 mL, 2.64 mmol, 2.5 eq.). The reaction was stirred at room temperature for 6 h. After removing the solvent under reduced pressure, the mixture was loaded onto a 25 g C18 cartridge and the product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 2 was obtained as a yellow powder (209 mg) with a yield of 76%. 1 H-NMR (400MHz, CD 3 OD): δ8.69(s, 1H), 7.94(d, J=7.92Hz, 1H), 7.57-7.51(m, 3H), 7.42-7.37(m, 1H), 4.21(s, 2H), 3.81(s, 3H). 13 C-NMR (100 MHz, CD 3OD): δ172.4, 160.9, 145.1, 143.7, 129.7, 129.4, 128.3, 121.8, 119.6, 112.4, 109.1, 56.8, 44.8.ESI-MS: m / z is C 13 H 13 N 2 O 4 [M+H] + Calculated for 261.3; findings 261.0.

[0267] Methyl (6-(3-((tert-butoxycarbonyl)amino)propoxy)quinoline-4-carbonyl)glycinate (3): Methyl (6-hydroxyquinoline-4-carbonyl)glycinate (2) (100 mg, 0.38 mmol, 1.0 eq.) and 3-(Bocamino)propyl bromide (230 mg, 0.96 mmol, 2.5 eq.) were dissolved in 4 mL of anhydrous DMF. 2 CO 3 (376 mg, 1.15 mmol, 3.0 eq.) was added to the solution and the reaction was stirred at room temperature overnight. After filtration, the solvent was removed under reduced pressure and the remaining mixture was loaded onto a 25 g C18 cartridge. The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 3 was obtained as a yellow powder with a yield of 54% (86 mg). 1 H-NMR (400 MHz, CDCl 3 ):δ8.68-8.37(m, 2H), 8.02(d, J=9.1Hz, 1H), 7.80(s, 1H), 7.72-7.64(m, 1H), 7.40(d, J=9.1Hz, 1H), 4.94(br s, 1H), 4.41-4.31(m, 2H), 4.27-4.18(m, 2H), 3.85(s, 3H), 3.44-3.30(m, 2H), 2.13-2.00(m, 2H), 1.43(s, 9H). 13 C NMR (100 MHz, CDCl 3 ):δ170.1, 167.2, 158.4, 144.7, 142.3, 128.4, 126.1, 124.7, 119.1, 103.7, 79.5, 60.4, 52.5, 41.4, 37.7, 29.3, 28.4.ESI-MS: m / z is C21 H 28 N 3 O 6 [M+H] + Calculated for 418.5; found 418.1.

[0268] tert-Butyl (S)-(3-((4-((2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)carbamate (5): Dissolve compound 3 (145 mg, 0.347 mmol, 1.0 eq.) and LiOH (58.4 mg, 1.39 mmol, 4.0 eq.) in 5 mL of H 2 The mixture was stirred in 0 / THF (1 / 1) for 6 h. After most of the THF was removed under reduced pressure, the mixture was loaded onto a 25 g C18 cartridge and eluted with a MeCN / water / TFA gradient (from 0 / 100 / 0.1 to 90 / 10 / 0.1) to remove salts. After removing the solvent and drying, the product 4 was obtained in quantitative yield. To a stirred solution of 4 (144 mg, 0.357 mmol, 1.0 eq.) in anhydrous DMF (4 mL) was added (S)-pyrrolidine-2-carbonitrile (56.8 mg, 0.428 mmol, 1.2 eq.), HATU (163 mg, 0.428 mmol, 1.2 eq.) and DIPEA (125 μL, 0.714 mmol, 2.0 eq.). After 6 h, the solvent was removed under reduced pressure and the crude mixture was loaded onto a 25 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 5 was obtained in 80% (135 mg) yield. 1 H NMR (400 MHz, CDCl 3):δ8.73(s, 1H), 7.95(d, J=10.2Hz, 1H), 7.68(br s, 1H), 7.63-7.56(m, 1H), 7.56-7.48(m, 1H), 7.38-7.29(m, 1H), 5.27(br s, 1H), 4.84-4.72(m, 1H), 4.46-4.35(m, 1H), 4.33-4.20(m, 1H), 4.17-4.09(m, 2H), 3.78-3.6 4(m, 1H), 3.59-3.46(m, 1H), 3.36(s, 2H), 2.38-2.17(m, 4H), 1.42(s, 9H), 1.35-1.27(m, 2H). 13 C NMR (100 MHz, CDCl 3 ):δ167.6, 167.5, 157.9, 156.2, 146.3, 130.2, 125.7, 123.7, 119.3, 118.0, 10 3.3, 79.0, 65.9, 46.8, 45.7, 42.2, 37.6, 29.8, 29.3, 28.4, 25.1.ESI-MS: m / z is C 25 H 32 N 5 O 5 [M+H] + Calculated for 482.6; findings 482.2.

[0269] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-6-(3-(pent-4-ynamido)propoxy)quinoline-4-carboxamide (FAP-acetylene 1): Compound 5 (100 mg, 0.207 mmol.) was treated with 1 mL of TFA / methylene chloride (1 / 1) for 2 h. The solvent was removed under reduced pressure and the crude mixture was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 6 was obtained in quantitative yield. To a stirred solution of 6 (10 mg, 0.026 mmol, 1.0 eq.) in DMF (1 mL) was added 2,5-dioxopyrrolidin-1-ylpent-4-ynoate (7) (5.12 mg, 0.026 mmol, 1.0 eq.) and DIPEA (13.5 μL, 0.078 mmol, 3.0 eq.). After 2 h, the solvent was removed under reduced pressure and the crude mixture was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, 8.6 mg of FAP-acetylene 1 was obtained in 71% yield. ESI-MS: m / z was C 25 H 28 N 5 O 4 [M+H] + Calculated 462.2; found 462.1.

[0270] (((S)-5-(5-amino-N-(4-bromobenzyl)pentanamido)-1-carboxypentyl)carbamoyl)-L-glutamic acid (9): To a stirred solution of 8 (214 mg, 0.326 mmol, 1.0 eq.) and 2,5-dioxopyrrolidin-1-yl 5-((tert-butoxycarbonyl)amino)pentanoate (102.5 mg, 0.326 mmol, 1.0 eq.) in DMF (2 mL) was added DIPEA (170 μL, 0.978 mmol, 3.0 eq.) at room temperature. The reaction mixture was stirred for 24 h and concentrated to give the crude product. The crude was diluted with 2 mL of TFA / CH2 Cl 2 (1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 9 (143 mg, 75%) was given as a white solid. H 1 -NMR (500MHz, D 2 O): δ7.55(d, J=8.5Hz, 1H), 7.51(d, J=8.5Hz, 1H), 7.12(dd, J=2.5, 8.0Hz, 2H), 4.59(s, 1H), 4.51(s, 1H), 4.29-4.22(m, 1H), 4.19-4.11(m, 1H), 3.39-3.29(m, 2H), 3.05-2.88(m, 2H), 2.59-2.38(m, 4H), 2.22-2 .10(m, 1H), 2.02-1.90(m, 1H), 1.82-1.46(m, 8H), 1.39-1.24(m, 2H).

[0271] (10S,23S,27S)-10-amino-18-(4-bromobenzyl)-2,2-dimethyl-4,11,17,25-tetraoxo-3-oxa-5,12,18,24,26-pentaazanonacosane-23,27,29-tricarboxylic acid (10): To a stirred solution of 9 (139 mg, 0.236 mmol, 1.0 eq.) and Fmoc-L-Lys(Boc)-OSu (133.9 mg, 0.236 mmol, 1.0 eq.) in DMF (2 mL) was added DIPEA (250 L, 1.42 mmol, 6.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product. To the above crude was added 2 mL of 20% piperidine in DMF (3 mL) at room temperature and the mixture was stirred for 1 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 10 (125 mg, 65%) was given as a white solid. 1-NMR (500MHz, DMSO-d 6 ):δ8.43-8.32(m, 1H), 8.08(s, 2H), 7.93(d, J=7.5Hz, 1H), 7.74(d, J=7.5Hz, 1H), 7.55 (d, J=8.0Hz, 1H), 7.41(d, J=8.0Hz, 1H), 6.74(t, J=5.0Hz, 1H), 6.40-6.26(m, 2H), 4.52 (s, 1H), 4.44(s, 1H), 4.14-3.98(m, 2H), 3.23-2.97(m, 5H), 2.93-2.82(m, 2H), 2.37(t , J=7.0Hz, 1H), 2.31-2.15(m, 3H), 1.96-1.43(m, 14H), 1.35(s, 9H), 1.29-1.15(m, 4H); [M+H] + is C 35 H 56 BrN 6 O 11 Calculated for (calcd) 815.3192; found for, 815.3184.

[0272] (11S,24S,28S)-1-Azido-19-(4-bromobenzyl)-11-(4-((tert-butoxycarbonyl)amino)butyl)-9,12,18,26-tetraoxo-3,6-dioxa-10,13,19,25,27-pentaazatriacontane-24,28,30-tricarboxylic acid (11): To a stirred solution of 10 (47.3 mg, 0.058 mmol, 1.0 eq.) and azido-PEG3-NHS ester (20 mg, 0.058 mmol, 1.0 eq.) in DMSO (100 μL) was added DIPEA (80 μL, 0.464 mmol, 8.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (from 0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 11 (42 mg, 70%) was given as a white solid. ESI-MS: m / z was C 44 H 71 BrN 9 O 15 [M+H] +Calculated 1044.4; findings 1044.3.

[0273] (14S,27S,31S)-22-(4-bromobenzyl)-14-(4-((tert-butoxycarbonyl)amino)butyl)-1-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazo (triazol-1-yl)-12,15,21,29-tetraoxo-3,6,9-trioxa-13,16,22,28,30-pentaazatritriacontane-27,31,33-tricarboxylic acid (12): A stirred solution of FAP-acetylene 1 (6 mg, 0.0129 mmol, 1.0 eq.) in t-BuOH:HO (1:2, 300 μL) was added with CuSO 4 .5H 2 HO (0.486 mg, 0.0019 mmol, 0.15 eq.) and sodium ascorbate (1.15 mg, 0.0058 mmol, 0.45 eq.) were added. To the above reaction mixture, a solution of product 11 (13.5 mg, 0.0129 mmol, 1.0 eq.) in DMSO (100 L) was added and stirred overnight at room temperature. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 12 (16.7 mg, 86%) was given as a white solid. ESI-MS: m / z was C 69 H 98 BrN 14 O 19 [M+H] + Calculated 1505.6; findings 1505.4.

[0274] (14S,27S,31S)-22-(4-bromobenzyl)-1-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)-12,15,21,29-tetraoxo-14-( 4-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetamido)butyl)-3,6,9-trioxa-13,16,22,28,30-pentaazatritriacontane-27,31,33-tricarboxylic acid (SB-FAP-01): Compound 12 (9 mg, 0.0059 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, it gave the free amine in quantitative yield. To a stirred solution of the amine (3.8 mg, 0.0027 mmol, 1.0 eq.) and DOTA-NHS-ester (3.08 mg, 0.004 mmol, 1.5 eq.) in DMSO (50 μL) was added DIPEA (4.7 μL, 0.027 mmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the crude was purified by preparative RP-HPLC chromatography using 0.1% TFA in HO and 0.1% TFA in acetonitrile as eluents followed by lyophilization to give compound SB-FAP-01 (3.3 mg, 69%) as a white solid. [RP-HPLC purification was achieved using an Agilent System, λ254 nm, 250 mmX10 mm Phenomenex Luna C18 column, solvent gradient: 90% HO (0.1% TFA) and 10% ACN (0.1% TFA), reaching 90% ACN in 25 min at a flow rate of 5 mL / min, product eluted at 13.8 min]. HRMS (ESI) m / z: [M+H] + is C 80 H 114 BrN 18 O 24 Calculated for, 1789.7456; found for, 1789.7442.

[0275] 1.2 Binding affinity (Ki) of SB-FAP-01 The binding affinity (Ki) of SB-FAP-01 is provided in Table 2.

[0276] [Table 2-1]

[0277] Example 2 Homobivalent FAP Ligands

[0278] In some embodiments, homobivalent FAP ligands can be prepared via the following scheme: [ka]

[0279] In some embodiments, the homobivalent FAP compound is selected from the group consisting of: [ka]

[0280] Example 3 Hetero-bivalent FAP ligands

[0281] In other embodiments, the heterobivalent FAP ligand includes a FAP-albumin heterobivalent compound having the following structure: [ka]

[0282] Example 4 Hetero-bivalent theranostics targeting FAP and PSMA 4.1 Overview

[0283] In some embodiments, the subject matter of the present disclosure provides theranostic radiopharmaceuticals that engage two major cell surface proteases, fibroblast activation protein alpha (FAP) and prostate specific membrane antigen (PSMA), each of which is frequently overexpressed within the tumor microenvironment (TME). The latter is also expressed in most prostate tumor epithelium. To engage in imaging and treatment of a broader spectrum of cancers, small molecule FAP and PSMA-targeting moieties have been 64 Conjugation was performed using an optimized linker to provide a Cu-labeled compound.

[0284] Representative compounds FP-L1 and FP-L2 were synthesized using two linker constructs with attached FAP and PSMA binding pharmacophores. In vitro inhibition constants (Ki) were determined for FAP and PSMA. Cellular uptake assays and flow cytometry were performed in human glioma (U87), melanoma (SK-MEL-24), and prostate cancer (PSMA+PC3 PIP and PSMA- PC3 flu) as well as clear cell renal cell carcinoma lines (PSMA+ / PSMA-786-O). Quantitative positron emission tomography / computed tomography (PET / CT) and tissue biodistribution studies were performed using experimental xenograft models of U87, SK-MEL-24, PSMA+PC3 PIP, and PSMA+786-O and the KPC genetically engineered mouse model of pancreatic cancer.

[0285] 64 Cu-FP-L1 and -L2 were produced with high radiochemical yields and molar activities. Ki values ​​were in the nanomolar range for both FAP and PSMA. PET imaging and biodistribution studies demonstrated that Cu-FP-L1 and -L2 are potent agonists against FAP and PSMA. 64 Cu-FP-L1 and 64 High and specific targeting of Cu-FP-L2 was demonstrated. 64 Cu-FP-L1 is 64 It showed better pharmacokinetics than Cu-FP-L2. 64 Cu-FP-L1 was detected after 2 hours in the U87 model. 64 It showed similar tumor uptake as Cu-FAPI-04. 64 Cu-FP-L1 demonstrated high tumor uptake and retention at >10% of the injected dose per gram of tissue from 1 to 24 hours post-injection in PSMA+PC3 PIP tumors.

[0286] To sum up, 64Cu-FP-L1 showed high and specific tumor targeting of FAP and PSMA. This compound should enable imaging of lesions expressing FAP, PSMA, or both on the tumor cell surface or within the TME. FP-L1 can be easily converted into a theranostic drug for managing heterogeneous tumors.

[0287] 4.2 Background

[0288] Theranostic radiopharmaceuticals are used to treat patients with metastatic cancer with high efficacy and low toxicity. Herrmann et al., 2020; Siva et al., 2020; Imlimthan et al., 2021; Uijen et al., 2021. Over the past decade, radiopharmaceutical development has focused on targeting selective cell surface receptors for specific biological targets, one target at a time. The "one molecule, one receptor" strategy has been quite successful. One successful low-molecular-weight radiotheranostic agent, 68 Ga- / 177 Lu-DOTATATE has received regulatory approval to treat somatostatin receptor-positive gastrointestinal pancreatic neuroendocrine tumors. In addition, several promising agents are underway, including prostate-specific membrane antigen (PSMA)-based radiopharmaceuticals. Herrmann et al., 2020.

[0289] Due to the heterogeneity of many cancers and their ability to rapidly acquire resistance, such highly selective agents may only provide temporary relief. Targeting appropriate cells in the tumor microenvironment (TME), which show differential expression in tumor cells, vasculature, and tumor stroma, may complement direct tumor targeting to enhance efficacy, especially when performed simultaneously. Bejarano et al., 2021. Without wishing to be bound by any particular theory, it was thought that heterobivalent agents targeting fibroblast activation protein alpha (FAP) and PSMA may enhance cancer detection and treatment, as both are abundantly expressed in the TME and the latter is expressed on prostate tumor cells. FAP is expressed on cancer-associated fibroblasts (CAFs) (Garin-Chesa et al., 1990), while PSMA is expressed on most prostate cancers and in solid tumor neovasculature. Chang et al., 1999; Wernicke et al., 2017; Spatz et al., 2018. Both FAP and PSMA are known to be present during disease progression in many invasive cancers and demonstrate increased expression in invasive and metastatic disease. Cohen et al., 2008; Busek et al., 2016; Lopez et al., 2016; Solano-Iturri et al., 2020a; Solano-Iturri et al., 2020b; Hofman et al., 2021.

[0290] PSMA-based radiotheranostics have proven beneficial in metastatic castration-resistant prostate cancer (mCRPC) compared with standard treatments. Hofman et al., 2021; Hofman et al., 2020. Patients with mCRPC have heterogeneous lesions and in some cases do not express PSMA. Paschalis et al., 2019. Lesions that are PSMA-negative, e.g., neuroendocrine prostate cancer (NEPC), can represent a particularly aggressive and often metabolically active disease. That fact has been exploited to select patients for PSMA-directed therapy by avoiding its use in patients with high 18F-fluorodeoxyglucose (FDG) uptake in their tumors. Paschalis et al., 2019.

[0291] Recent immunohistochemistry (IHC) studies have further begun to reveal that FAP expression is a hallmark of mCRPC, regardless of genetic subtype, treatment regimen, or metastatic location. Hintz et al., 2020; Kesch et al., 2021. Recent studies have also shown that FAP-based PET imaging is more sensitive for detecting PSMA-negative metastatic lesions than FDG PET / CT. Isik et al., 2021; Kessel et al., 2021. FAP-based PET imaging has emerged as a new diagnostic tool in various malignancies. Kratochwil et al., 2019; Mona et al., 2021. FAP is a complex membrane protease overexpressed on CAFs in >90% of human epithelial tumors. Kalluri, 2016. It is also an independent negative prognostic factor for several malignancies (Fitzgerald and Weiner, 2020; and exists on the cell surface and in soluble, circulating form in the blood in mice and humans (Keane et al., 2014). CAFs produce cytokines, chemokines, metabolites, enzymes, and extracellular matrix molecules that play a key role in fueling cancer cell proliferation (Kalluri, 2016). Like PSMA, FAP is a promising clinical drug candidate. 68Ga-FAPI-04 and 68 Employing high affinity inhibitors, including Ga-FAPI-46 (Figure 9), Brennen et al., 2012, allows selective targeting of various tumors. Loktev et al., 2019. Like FAP, PSMA is also a protease known as glutamic acid carboxypeptidase II (GCP II) and increases endothelial cell invasion and angiogenesis in most invasive solid tumors. Conway et al., 2013.

[0292] Copper-64 [Half-life (t 1 / 2 ) = 12.7 hours; β+: 17.4%, Eβ+ (average) = 278 keV] has a unique decay profile and is primarily used for PET imaging, 67 Cu[t 1 / 2 = 61.9 hours, β- = 100%, Eβ- (average) = 141 keV] is a promising radioisotope for therapy. Copper-64 is an attractive alternative to short-lived radioisotopes for PET imaging of antibodies, peptides, and small molecules. Ling et al., 2019. Recent 64 Cu-DOTATATE (LUTATHERA (R) )(Lutathera, (R) Regulatory approval of 1,2,3-Trifluoroethylene (1,2-difluoroethylene) for several molecular targeted radiotheranostic applications. 64 This prompted the development of Cu-labeled compounds. Ling et al., 2019.

[0293] Without wishing to be bound by any particular theory, it was believed that heterobivalent compounds using two clinically tested high affinity FAP and PSMA-based targeting moieties could combine and enable imaging and treatment of various cancers and cancer subtypes within a given malignancy, such as PSMA+ mCRPC and PSMANEPC. Such compounds could also enhance retention of PSMA-based radiotheranostics in solid malignancies, such as glioblastoma, that have PSMA+ neovasculature and FAP+ tumor cells or CAFs. Uijen et al., 2021; Busek et al., 2016; Rohrich et al., 2019. As a first step in testing this hypothesis, optimized heterobivalent compounds with two distinct binding pharmacophores targeting FAP and PSMA were developed.

[0294] Previously optimized small molecule FAP targeting moieties, Jansen et al., 2014; Slania et al., 2021, and PSMA targeting platforms were used to generate high affinity heterobivalent compounds. The compounds were 64 It was labeled with Cu and evaluated in preclinical imaging studies in relevant human xenografts and in the KPC genetically engineered mouse model (GEMM) of pancreatic ductal adenocarcinoma (PDAC).

[0295] 4.3 Materials and Methods 4.3.1 Reagents, cell lines, and animal models

[0296] Six cell lines were used for in vitro and in vivo evaluation: U87 (glioblastoma), SK-MEL-24 (melanoma), PSMA+PC3 PIP, PSMAPC3 flu (prostate cancer), PSMA+786-O, and PSMA-786-O (renal cell carcinoma). Six to eight-week-old male, nonobese diabetic / shi-scid / IL-2rγ(null) (NSG) mice (Johns Hopkins Animal Resources Core) were implanted subcutaneously with the indicated cell lines.

[0297] 4.3.2 In vitro assays 4.3.2.1 Competitive inhibition assay

[0298] The PSMA binding affinity of the compounds was determined using a competitive inhibition assay as previously reported. Banerjee et al. FAP, prolyl endopeptidase (PREP), and dipeptidyl dipeptidase 4 (DPPIV) inhibition assays: Recombinant enzymes (FAP, PREP, DPPIV) were purchased from R&D Systems (Minneapolis, MN). FAPI-04 was used as a positive control. Z-Gly-Pro-AMC was used as a substrate for FAP and PREP. H-Gly-Pro-AMC was used as a substrate for DPPIV. Recombinant enzymes (0.4 μg / mL) were incubated with various amounts of test articles in the presence of the indicated substrates (80 μM) for 10 min at room temperature. Fluorescence intensity was measured with an excitation of 380 nm and emission of 460 nm using a Cytation 5 Cell Imaging Multi-Mode Reader (BioTek, Winooski, VT). IC50 and Ki values ​​were determined using a sigmoidal dose-response function. Cheng and Prusoff, 1973.

[0299] 4.3.2.2 Cellular uptake, flow cytometry, and IHC

[0300] Cellular uptake assays, flow cytometry, and IHC were performed as previously described (Banerjee et al., 2019; Nimmagadda et al., 2018). PET Imaging and Biodistribution To quantify and validate the PET imaging data, sequential PET imaging and biodistribution studies were performed. Briefly, tumor-bearing mice were administered approximately 7.4 MBq of radiotracer in 150 μL of saline via tail vein injection. They were randomized into the indicated groups of 3-4 mice before injection with the radiotracer.

[0301] In experiment 1, bilateral xenografts of U87 tumors (right flank) and PSMA+PC3 PIP tumors (left flank) were established in male NSG mice (n=3) that were followed for imaging followed by biodistribution over 24 hours. Blocking studies were performed with separate cohorts of U87 and PSMA+PC3 PIP bilateral xenografts (n=3-4) by co-injection of FAPI-04 (for FAP), Loktev et al., 2019, or ZJ43 (for PSMA), Olszewski et al., 2004, to demonstrate FAP or PSMA binding specificity. 64 Cu-FP-L1 and 64 To compare the tissue distribution characteristics of Cu-FAPI-04, biodistribution studies were further performed using U87 tumors (n=3) 2 h post-injection.

[0302] In experiment 2, SK-MEL-24 tumor-bearing mice (n=3-4) were examined.

[0303] In experiment 3, PET studies were performed at 1 and 2 h after injection using GEMMs, [6-mo-old, male, LSL-KrasG12D;LSL-Trp53R173H;Pdx1-Cre(KPC) triple mutant], He et al., 2020. PET studies were performed simultaneously with age-matched littermates (female). After 48 h, mice were injected with a near-infrared fluorescent (NIRF) compound, IRDye800-FP-L1, which contains the same construct as FP-L1 for FAP and PSMA targeting, with the 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) chelator replaced by IRDye800CW (LICOR, Lincoln, NE). Mice were then sacrificed at 2 h, and ex vivo optical imaging studies were performed. A veterinary pathologist (Dr. Brayton) performed the histopathological evaluation.

[0304] Experiment 4 consisted of exploratory imaging (7.4MBq) (n=2) and 2-hour biodistribution studies (0.74MBq) in male NSG healthy mice (n=3) and tumor-bearing human clear cell renal cell carcinoma (ccRCC) PSMA+786-O tumors (n=4).

[0305] 4.3.3 Statistical analysis

[0306] Statistical significance was calculated using unpaired two-tailed t-tests with GraphPad Prism 9.0 software. Data are expressed as mean ± standard deviation (± SD). Statistical significance was defined as P ≤ 0.05.

[0307] 4.4 Results 4.4.1 Chemical and radiochemical synthesis

[0308] The structures of compounds FP-L1 and FP-L2 are shown in Figure 9. Click chemistry was used to conjugate the FAP and PSMA targeting moieties with the selected linker. Radiolabeling was achieved in nearly quantitative yields. 64A rapid microwave-assisted method was employed to generate Cu-FP-L1, and high-performance liquid chromatography (HPLC) was used to remove unreacted ligand and ensure high specific activity (>19 MBq / nmol). 64 Cu-FP-L1 was stable in PBS and bovine serum albumin at 37° C. for at least 4 h.

[0309] 4.4.2 In vitro characterization 4.4.2.1 Inhibitory potential

[0310] FP-L1 and FP-L2 showed high binding affinity to FAP (Table 2, FIG. 17), comparable to that of FAPI-04, studied in the same assay as the reference compound. The PSMA binding affinity of FP-L1, carrying a polyethylene glycol (PEG) linker, was 2-fold lower than that of FP-L2, carrying a pentamethylene linker. These values ​​were lower than ZJ43, Olszewski et al., 2004, a known PSMA-binding compound used as a standard for the assay.

[0311] [Table 2-2]

[0312] 4.4.2.2 Cellular uptake

[0313] 64 Cu-FP-L1 showed the highest uptake in FAP+U87 and SK-MEL-24 cells compared to PSMA+PIP and PSMA-PC3 flu and 786 PSMA+ / PSMA- cells (Figure 10A). Blocking studies revealed a significant (P<0.001) reduction in uptake by co-incubation with FAPI-04 (FAP blocking) or ZJ43 (PSMA blocking), demonstrating the binding specificity of the compounds. Cellular uptake was proportional to the surface expression of the target protein in the studied cell lines as revealed by flow cytometry (Figure 10B).

[0314] 4.4.3 In vivo characterization 4.4.3.1 Biodistribution and PET Imaging (Experiment 1)

[0315] 64 Cu-FP-L1 was evaluated in a mouse xenograft model bearing both PSMA+ human prostate cancer PC3 PIP and FAP+ U87 tumors in the left and right upper flank, respectively (Figure 11A, Figure 11B). Biodistribution data (Figure 11C) revealed high tumor uptake in FAP+ U87 tumors (16.96 ± 5.01% ID / g at 2 h, 19.05 ± 5.89% ID / g at 4 h, and 4.31 ± 0.75% ID / g at 24 h). Also, uptake in PSMA+ PC3 PIP tumors remained high at 12.06 ± 0.78% ID / g at 2 h, 18.89 ± 3.95% ID / g at 4 h, and 10.51 ± 1.82% ID / g at 24 h). Previously reported FAP-based compounds, 111 Compared to In-QCP02 (Slania et al., 2021), 64 Tumor uptake and retention of Cu-FP-L1 was higher from 2 h post-injection onwards, whereas nonspecific healthy tissue clearance was comparable. Blocking studies to determine binding specificity with FAPI-04 (500 nmol / kg) (Figure 11E) showed a significant decrease in uptake in U87 tumors (2.47 ± 0.82%ID / g) and an increase in uptake in PSMA+PC3 PIP tumors (18.56 ± 2.02%ID / g) after 2 h. A significant decrease in activity was observed in healthy tissues, especially blood, salivary gland and lacrimal gland. The relatively high bone uptake (>1%ID / g) was consistent with the previous study. 64This may be related to the initial high blood and bone marrow uptake of Cu-FP-L1. In contrast, PSMA blockade with ZJ43 (500 nmol / kg) caused significantly lower uptake in PSMA+PIP tumors (7.48±0.5%ID / g), pointing to the PSMA specificity of the drug. Specificity was further supported by a >2-fold lower uptake in the kidney (2.97±0.23%ID / g) compared with unblocked drug (8.03±0.89%ID / g), a known site of endogenous PSMA expression. Silver et al., 1997.

[0316] This model (FAP+U87 and PSMA+PC3 PIP) 64 The drug was evaluated using Cu-FP-L2. 64 Uptake in the kidney was significantly higher than Cu-FP-L1 (FIG. 19) and uptake in the tumor was relatively low and was not studied further. 64 Biodistribution studies with Cu-FP-L1 also demonstrated that small tumors (approximately 51.9 ± 16.5 mm) were significantly increased after 2 h of injection. 3 ), and 64 Compared with Cu-FAPI-04. Tumor uptake was comparable. 64 10.84 ± 1.02%ID / g for Cu-FP-L1, and 64 For Cu-FAPI-04, the value was 9.53 ± 2.55% ID / g (Figure 11F). 64 Cu-FP-L1 was high, but uptake in the salivary glands was 64 Cu-FAPI-04 was higher. IHC studies verified high FAP and absent / low PSMA expression in U87 tumors, and relatively low FAP and high PSMA expression in PSMA+PC3 PIP tumors (Figure 11G). Imaging and biodistribution in SK-MEL-24 human melanoma model (Experiment 2). 64Cu-FPL1 was evaluated in FAP+SK-MEL-24 tumor xenografts (n=4, Fig. 4A-4E). Strong tumor accumulation was observed 15 min after injection, but activity disappeared over the next 2 h. Among healthy tissues, kidney (9.71±2.48%ID / g), liver (3.54±0.46%ID / g), salivary gland (3.77±0.35%ID / g) and bone (2.91±0.59%ID / g) showed relatively high uptake. At 24 h after injection, no detectable tumor accumulation was observed. Uptake in other healthy tissues was within the range of that of blood (1.51±0.15%ID / g). Receptor blocking studies with FAPI-04 and ZJ43, and dual blockade (self-blockade) with FP-L1 were performed (Fig. 12C). Blocking studies were monitored by PET and showed significantly lower tumor uptake for FAP blockade and dual FAP and PSMA blockade compared to PSMA blockade. PSMA blockade was evident by reduced signal intensity in the kidney, a known PSMA-expressing tissue. SK-MEL-24 express medium-low PSMA, but PSMA levels in these cells were previously shown to be approximately 10-fold lower compared to PSMA+PC3 PIP tumors (Nimmagadda et al., 2018); however, the study cohort of tumors showed low PSMA expression as revealed in the IHC study (Figure 12F).

[0317] 4.4.3.2 Imaging of pancreatic cancer in GEMM (Experiment 3)

[0318] 64Imaging of Cu-FP-L1 was performed in immunodeficient KPC mice (Figure 13). Pancreatic ductal carcinoma (PDAC) progression in KPC mice recapitulates human PDAC. He et al., 2020. PET demonstrated higher uptake in the abdominal area 2 hours after injection in KPC mice than in healthy mice (Figure 13B). To confirm the distribution of the radiotracer, mice were injected with the corresponding NIRF contrast agent, IRDye800-FP-L1 (5 nmol) (Figure 20), carrying the same heterobivalent construct as FP-L1, followed by NIRF imaging of dissected organs 2 hours later. As shown in Figure 13C, IRDye800-FP-L1 specifically accumulated in the pancreas of KPC mice, demonstrating a clear margin between it and the healthy spleen. In contrast, the pancreas of control mice showed minimal uptake. Both mice showed high uptake in the kidney, but due to renal clearance. Histopathological evaluation further confirmed the presence of PDAC lesions. PDAC lesions were <2 mm and were associated with high FAP expression as confirmed by IHC (Figure 13D). IHC data also revealed moderate PSMA expression within the TME (Figure 13D). Lung metastases (<1 mm) were also observed, suggesting the high sensitivity of the radiotracer (histology, Figure 21). 64 Cu-FP-L1 and IRDye800-FP-L1 can delineate pancreatic tumors and metastases by PET and NIRF imaging, respectively, in the KPC model of PDAC.

[0319] 4.4.3.3 Imaging of ccRCC in PSMA+786-O (Experiment 4)

[0320] FAP expression is associated with tumor aggressiveness and reduced survival in ccRCC. Lopez et al., 2016. Lower concentrations of soluble FAP in the plasma of patients with ccRCC compared to healthy controls predict tumor progression. Solano-Iturri et al., 2020. A PSMA+786-O tumor model that retains the ccRCC phenotype was recently developed to test these clinical events. This model showed lower FAP expression compared to U87 and SK-MEL-24, and lower PSMA expression compared to PSMA+PC3 PIP tumors, as revealed by flow cytometry (Figure 10B). In NSG mice bearing PSMA+ 786-O xenografts and healthy NSG mice 64 Head-to-head biodistribution performed with Cu-FP-L1 demonstrated high renal uptake compared to healthy mice (Figure 14B). High tumor uptake was also observed in PET imaging and biodistribution studies after 2 hours (11.97±1.63%ID / g). IHC revealed high PSMA and FAP expression within tumor sections (Figure 14C).

[0321] 4.3 Discussion

[0322] One goal of the subject matter of this disclosure was to develop a dual-targeting radioligand that could detect a broader range of cancers than is possible with current agents. Because FAP and PSMA are medically important as cancer biomarkers and because they are expressed in many cancers, it was chosen to target these two cell surface proteins with the same compound. It was also interesting to have one versatile agent that would allow for the detection and treatment of a wide range of prostate cancers, from castration sensitivity to mCRPC, including NEPC (the latter does not express PSMA, but has been shown to be detectable with 68Ga-FAPI-04). Kesch et al., 2021. Compared to the administration of a cocktail containing radioactive FAP and PSMA-targeted compounds, a single agent is likely to provide more uniform dosimetry and pharmacokinetics, and thus may be clinically easier to interpret. It is noted that it is not necessarily expected or intended that these heterobivalent compounds bind both FAP and PSMA simultaneously. FAP and PSMA reside in different cells and tissues, within the TME, or, in the case of PSMA and prostate cancer, within the tumor epithelium.

[0323] Reprogramming the TME by targeting specific, contributing cells, e.g., CAFs, macrophages or T-cell subtypes, is a new and promising approach for cancer treatment and overcoming resistance. Dual targeting approaches have the advantage of managing two biologically distinct targets and could result in synergistic effects. In certain cancers, it is beneficial to engage therapeutic radiopharmaceuticals both in the associated CAFs as well as in neovasculature. One such example is clear cell renal cell carcinoma (ccRCC), where FAP expression correlates with more aggressive and metastatic disease as cancer cells undergo epithelial to mesenchymal transition. Errarte et al., 2016.ccRCC has been shown to be imaged to good advantage with the chimeric neovasculature of ccRCC, where tumor vessels are composed of both endothelial and cancer cells, using PSMA-targeted PET agents, Meyer et al., 2019; Delgado-Bellido et al., 2017; Zhou et al., 2016. Antivascular agents are used to treat ccRCC, more recently in combination with immune checkpoint inhibitors, Rini et al., 2019, and targeting of neovasculature - by radioactive theranostics - may allow tumor growth control. Other such cancers, which highly express FAP and PSMA, Silania et al., 2021; Nimmagadda et al., 2018; Pure and Blomberg, 2018, and for which a dual targeting approach may prove useful, include melanoma, breast, glioma, lung, ovarian, upper aerodigestive tract cancer, and pancreatic, the last of which was demonstrated in KPC mice (Figure 13).KPC mice were chosen because pancreatic cancer is known to express both FAP and PSMA (Stock et al., 2017; Pereira et al., 2019; Poels et al., 2021; Krishnaraju et al., 2021) and is notoriously difficult to image. The dual targeting approach also provides an early stage mechanism when one of the targets is pharmacokinetically inaccessible in a particular lesion or expressed at a different disease stage than the other. Examples of the strategy include heterobivalent, bispecific antibodies that have received regulatory approval for the treatment of certain cancers (Sheridan, 2015); heterobivalent immunoligands have also demonstrated enhanced tumor uptake and retention in PET (Luo et al., 2015). The subject matter of the present disclosure also employed a heterobivalent approach, however, focusing on small molecule targeting moieties. Such a strategy is possible by targeting PSMA and integrin αvβ3, which has previously been shown to provide synergistic effects in targeting tumor neovasculature (Shallal et al., 2014). Others have similarly attempted dual targeting of PSMA and other cell surface proteins, notably gastrin-releasing peptide receptor (GRPR) (Bandari et al., 2021; Eder et al., 2014).

[0324] The subject matter of the present disclosure, in some embodiments, provides two orthogonal targets, one on CAFs (FAP) and the other on neovasculature (PSMA), to provide a synergistic effect, while also allowing target engagement by theranostic agents when one target is missing from the lesion. 64 To test the ability of Cu-FP-L1 to engage FAP and PSMA at the same dose, we sought tumor cell lines known to have at least moderate expression of both targets, such as SK-MEL-24 ( Nimmagadda et al., 2018 ).

[0325] However, synergistic uptake was not observed, i.e., in that cell line 64This was consistent with the IHC data showing low PSMA in the study cohort of tumors, with Cu-FP-L1 engaging both FAP and PSMA (Figure 10) or with in vivo targeting (Figure 12). Notably, in PC3 PIP tumors 64 The in vivo tumor targeting of Cu-FP-L1 was significantly higher than that reported for PSMA-based heterobivalent compounds. Bandari et al., 2021. 64 The biodistribution data for Cu-FAPI-04 demonstrated similar in vivo properties as reported in Watabe et al., 2020. Also, higher tumor uptake (%ID / g) was observed in tumors with volumes <100mm in the U87 model. 3 compared with tumors of 200 mm 3 It should be noted that the large tumor volume was associated with tumor volumes greater than 100 μg / kg. Without wishing to be bound by any particular theory, it is believed that the large tumors likely result in the incorporation of additional stromal or desmoplastic tissue into the TME.

[0326] From these studies, 64 These results suggest that Cu-FP-L1 can detect both FAP+ and PSMA+ tumors in vivo while minimizing nonspecific tissue accumulation. In addition to detecting FAP and PSMA in the TME of various solid tumors, 64 Cu-FP-L1 or a suitable analogue could enable imaging of a spectrum of prostate cancer subtypes, including those that no longer express PSMA. Isik et al., 2021. Although more lesions would be detected, a limitation of this approach is that it is not known whether lesions were detected based on FAP or by expression of PSMA (or both), thus necessitating tissue sampling for definitive characterization. This limitation is likely offset by increased sensitivity. 64Agents similar to Cu-FP-L1 could be used to get a fuller picture of the lesions present in patients being staged for prostate cancer, or to follow up patients undergoing PSMA-specific radiopharmaceutical therapy to understand why they may not be responding, for example through the localization of appearing neuroendocrine-differentiated lesions. The corresponding radiotherapy, 67Cu-FP-L1, has the potential to treat both PSMA+ and NEPC cancers simultaneously.

[0327] 4.6 Overview

[0328] The data disclosed herein are 64 We show that Cu-FP-L1 can target both PSMA and FAP expression in the same in vivo experiment. By targeting two widely used targets, FAP, which has been noted as a pan-cancer marker, 64 Cu-FP-L1 has the potential to detect more than one type of cell on the tumor cell surface or in the TME, and corresponding treatments may address resistance issues due to tumor heterogeneity.

[0329] Example 5 Fibroblast activation protein alpha (FAP-α)-targeted imaging and radiation therapy 5.1 Overview

[0330] Fibroblast activation protein-α (FAP-α) expression has been detected on the surface of fibroblasts in the stroma surrounding >90% of epithelial cancers (malignant breast, colorectal, skin, prostate, pancreatic cancer, and the like) and inflammatory diseases (arthritis, fibrosis, and the like), and is rarely expressed in healthy tissues. Imaging and radiotherapeutic agents that specifically target FAP-α are of clinical importance. FAP is similar in sequence and catalytic domain to dipeptidyl peptidase IV (DPPIV) and propyl endopeptidase (PREP), so most inhibitors of FAP also bind to DPPIV and PREP. Here, we developed potent and selective low molecular weight (LMW) inhibitors of FAP-α bearing targeting moieties amenable to modification with radiolabels and optical groups. These compounds allow for in vivo nuclear imaging (such as PET and SPECT), optical imaging and radiotherapy targeting of FAP-α. The targeting moieties of the present disclosure can be adapted to other optical dyes and radioisotopes for imaging and therapeutic applications targeting FAP-α.

[0331] In part, the subject matter of the present disclosure demonstrates that: (1) radiolabeled isotopes with sufficient half-life conjugated to low molecular weight (LMW) FAP-α selective inhibitors can enable successful in vivo and ex vivo imaging; (2) the disclosed compounds are useful for, but not limited to, FAP-α related imaging applications. 18 F, 68 Ga, 64 Cu, 86 Y, 90 Y, 89 Zr, 111 In, 99m Tc, 125 I, 124 I, 11 C. 76(3) the compounds of the present disclosure are useful for, but are not limited to, FAP-α-related radiation therapy, including, but not limited to, the use of optical dyes, PET and SPECT isotopes; 90 Y, 177 Lu, 125 I, 131 I, 211 At, 111 In, 153 Sm, 186 Re, 188 Re, 67 Cu, 212 Pb, 225 Ac, 213 Bi, 212 Bi, 212 Pb, 67 Ga, 77 Br, 161 It further allows for radiolabeling with radiotherapeutic isotopes, including Tb; compounds of the present disclosure showed specific binding to FAP-positive cell lines over the closely related peptidases DPPIV and PREP.

[0332] 5.2 Background 5.2.1 Fibroblast activation protein alpha (FAP-α)

[0333] FAP-α expression is associated with the tumor microenvironment and has been detected on the surface of stromal fibroblasts surrounding over 90% of epithelial-derived cancers and their metastases. Garin-Chesa et al., 1990; Rettig et al., 1993; Tuxhorn et al., 2002; Scanlan et al., 1994. FAP-α plays an important role in promoting angiogenesis, proliferation, invasion, and suppression of tumor cell death. Allinen et al., 2004; Franco et al., 2010. In healthy adult tissues, FAP-α expression is only restricted to areas of tissue remodeling or wound healing. Scanlan et al., 1994; Yu et al., 2010; Bae et al., 2008; Kraman et al., 2010.

[0334] FAP expression is extremely difficult to detect in non-diseased adult organs, but is greatly elevated at sites of tissue remodeling, including lung or liver fibrosis, arthritis, and tumors (Scanlan et al., 1994; Yu et al., 2010). These characteristics make FAP-α an excellent imaging and radiotherapy target for cancer and inflammatory diseases.

[0335] 5.2.2 Implications for FAP-α imaging with low molecular weight (LMW) agents.

[0336] FAP-α is expressed in the tumor stroma, and anti-FAP antibodies have been investigated for radioimmunotargeting of malignancies, including murine F19, sibrotuzumab (a humanized version of the F19 antibody), ESC11, and ESC14, among others. Welt et al., 1994; Scott et al., 2003; Fischer et al., 2012. Antibodies have also demonstrated the potential to image inflammation, such as rheumatoid arthritis. Laverman et al., 2015. However, antibodies as molecular imaging agents suffer from pharmacokinetic limitations, including slow blood and non-target tissue clearance (usually 2-5 days or longer) and non-specific organ uptake. LMW agents demonstrate faster pharmacokinetics and higher specific signals within clinically convenient times after administration. They are also more easily synthesized in radiolabeled form, providing a shorter route to regulatory approval. Coenen et al., 2010; Cho et al., 2012; Reilly et al., 2015.

[0337] FAP-α are type II transmembrane serine proteases of the prolyl oligopeptidase family, which are distinguished by their ability to cleave post-proline peptide bonds. They have been shown to play a role in cancer by modifying bioactive signaling peptides through this enzymatic activity. Kelly, 2005; Edosada et al., 2006. FAP-α exists as a homodimer and performs its enzymatic function. Inhibitors that selectively target FAP-α have been reported. Lo et al., 2009; Tsai et al., 2010; Ryabtsova et al., 2012; Poplawski et al., 2013; Jansen et al., 2013; Jansen et al., 2014. DPPIV is the closest homolog of FAP, with over 50% sequence similarity and over 70% similarity in the catalytic region. Juillerat-Jeanneret et al., 2017. Prolyl endopeptidase (PREP) is phylogenetically related to FAP and cleaves the post-proline peptide bond of its substrates similar to FAP (Jambunathan et al., 2012). Therefore, it is essential to establish the specificity of imaging compounds for FAP rather than DPPIV and PREP. FAP-α selective inhibitors could potentially provide a direct optical agent or radioisotope solution for FAP-α targeted imaging and therapy.

[0338] The subject matter of the present disclosure provides FAP-α selective targeting moieties that are amenable to modification with optical dyes, radioactive metal chelate complexes, and other radioisotope prosthetic groups, providing a platform for imaging and radiotherapy targeted to FAP-α. In some instances, the FAP pharmacophore and labeled prosthetic groups, e.g., 18 The linker is chosen so that the F-labeled prosthetic group is separated by a hydrophilic amino acid-based linker. The hydrophilic linker reduces nonspecific binding and improves the pharmacokinetics of the radiotracer.

[0339] 5.2.3 Implications for nuclear imaging and radiotherapy

[0340] Radionuclide molecular imaging, including PET, is the most mature molecular imaging technique without the limitation of tissue penetration. Due to its advantages of high sensitivity and quantification, radionuclide molecular imaging plays an important role in clinical and preclinical research. Youn and Hong, 2012; Chen et al., 2014.

[0341] Many radionuclides, mainly beta- and alpha-emitters, have been investigated for targeted radioimmunotherapy, and include both radioactive halides and radioactive metals (Table 1). Highly potent and specific binding moieties targeting FAP-α would enable its nuclear imaging and radiotherapy. Here, we describe the first synthesis of a nuclear imaging and radiotherapy agent based on this dual targeting moiety against FAP-α for nuclear imaging.

[0342] 5.3 Experimental section 5.3.1 Ligand synthesis

[0343] [ka] Scheme 1. Synthesis of JHU1220627.

[0344] 6-(3-((tert-butoxycarbonyl)amino)propoxy)quinoline-4-carboxylic acid (2): To a stirred solution of 6-hydroxyquinoline-4-carboxylic acid (1) (300 mg, 1.586 mmol, 1 eq.) and cesium carbonate (1.55 g, 4.758 mmol, 3 eq.) in 5 mL of DMF, tert-butyl (3-bromopropyl)carbamate (944 mg, 3.965 mmol, 2.5 eq.) was added and the mixture was allowed to stir at 60° C. overnight. The mixture was cooled to room temperature and diluted with 2 mL of acetonitrile and 5 mL of water followed by the addition of 600 μL of 12 M NaOH. After stirring for 30 min at room temperature, the reaction mixture was loaded onto a 30 g C18 cartridge (Biotage Sfar). The product was purified with a MeCN / water / TFA gradient (from 5 / 100 / 0.1 to 60 / 10 / 0.1). 312 mg of product 2 was obtained in 90% yield. 1 H-NMR (400MHz, CD 3 OD): δ8.80(d, J=3.2Hz, 1H), 8.28(s, 1H), 8.04-7.98(comp, 2H), 7.51(d, J=7.2Hz, 1 H), 4.19(t, J=7.6Hz, 2H), 3.29-3.26(comp, 2H), 2.04(t, J=7.6Hz, 2H), 1.42(s, 9H). ESI-MS: m / z is C 18 H 23 N 2 O 5 [M+H] + Calculated 347.4; found 347.1.

[0345] (S)-6-(3-aminopropoxy)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)quinoline-4-carboxamide (4): To a stirred solution of 2 (184 mg, 0.53 mmol, 1.0 eq.), 3 (122 mg, 0.795 mmol, 1.5 eq.) and HATU (302 mg, 0.795 mmol, 1.5 eq.) in DMF (3 mL) was added DIPEA (231 μL, 1.325 mmol, 2.5 eq.) at room temperature. The reaction mixture was stirred overnight at room temperature. After removal of the solvent, 2 mL of TFA / CH was added under reduced pressure.2 Cl 2 (1:1) was added and stirred for 30 min. The reaction mixture was concentrated and loaded onto a 30 g C18 cartridge (Biotage Sfar). The product was purified by MeCN / water / TFA gradient (5 / 100 / 0.1 to 60 / 10 / 0.1). After lyophilization, 120 mg of product 4 was obtained in 74% yield. NMR (400 MHz, CD 3 OD): δ8.93(d, J=4.0Hz, 1H), 8.17(d, J=2.0Hz, 1H), 8.10(d, J=7.2Hz, 1H), 7.81(d, J=4.0Hz, 1H), 7.67(dd, J=7.6, 2.0Hz, 1H), 4.84-4.78 (m, 1H), 4.47-4.38(comp, 2H), 4.38-4.28(comp, 2H), 3.86-3.78(m, 1H), 3.70-3.62(m, 1H), 3.24-3.18(comp, 2H), 2.39-2.18(comp, 6H). ESI-MS: m / z is C 20 H 24 N 5 O 3 [M+H] + Calculated 382.4; found 382.2.

[0346] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-6-(3-((4-iodobenzyl)amino)propoxy)quinoline-4-carboxamide (5): Dissolve 4 (40 mg, 0.105 mmol, 1.0 eq.) and 4-iodobenzaldehyde (27 mg, 0.115 mmol, 1.1 eq.) in 2 mL of 1,2-dichloroethane / CH 3 OH (1:1), sodium triacetoxyborohydride (76 mg, 0.314 mmol, 3 eq.) were added and stirred at 60° C. for 48 h. After cooling to room temperature, the reaction mixture was diluted with 1% to 15% CH 3 CH in OH 2 Cl 2 The product was purified by silica gel flash chromatography eluting with 50% hexanes. 23 mg of product 4 was obtained in 50% yield. NMR (400 MHz, CD 3OD): δ8.80-8.75(m, 1H), 8.02-7.96(m, 2H), 7.80(d, J=6.00Hz, 2H), 7.58(d, J=3.60Hz, 1H), 7.45(dd, J=7.20, 1.6Hz, 1H), 7.26(d, J=6.40Hz, 2H), 4.81-4 .75(m, 1H), 4.40-4.34(comp, 2H), 4.34-4.28(comp, 2H), 4.15(s, 2H), 3.86- 3.77(m, 1H), 3.70-3.61(m, 1H), 3.28-3.21(comp, 2H), 2.38-2.17(comp, 6H). ESI-MS: m / z is C 27 H 28 IN 5 O 3 [M+H] + Calculated 598.5; found 598.1.

[0347] Compound JHU1220627: To a stirred solution of 5 (7.5 mg, 11.41 μmol, 1.0 eq.) and DOTA-NHS-ester (10.4 mg, 13.69 μmol, 1.2 eq.) in DMF (250 μL) was added DIPEA (20 μL, 228.13 μmol, 20 eq.) at room temperature. The reaction mixture was stirred at 70° C. for 2 h and another 1.2 eq. of DOTA-NHS ester was added and stirred overnight. The solvent was then evaporated and redissolved in acetonitrile / water (1:1) and the pH was adjusted to approximately 3 by the addition of TFA. The product was purified by preparative HPLC chromatography using a Phenomenex 21.2mmX250mm Luna, λ254nm, 10μm PREP C18(2) column and isocratic eluent 30% acetonitrile / 70% water / 0.1% TFA at a flow rate of 10mL / min. After lyophilization, 6.2mg of JHU1220627 was obtained as a white solid in 55% yield. ESI-MS: m / z was C 43 H 55 IN 9 O 10 [M+H] + Calculated for 984.9; found 984.3.

[0348] [ka] Scheme 2. Synthesis of JHU1221785.

[0349] Compound 7: To a stirred solution of 4 (120 mg, 0.3146 mmol, 1.0 eq.), Fmoc-D-Asp(t-Bu)-OH (388 mg, 0.9438 mmol, 3 eq.) and HATU (359 mg, 0.9438 mmol, 3 eq.) in DMF (3 mL), DIPEA (219 μL, 1.2584 mmol, 4 eq.) was added and the mixture was allowed to stir at room temperature overnight. After evaporating the solvent, the mixture was stirred in 20% piperidine in DMF for a period of 30 min. The reaction mixture was then diluted with EtOAc and washed with water (3x) and brine. The combined organics were washed with anhydrous Na 2 SO 4 The mixture was dried over hexane, filtered, concentrated, and dried under reduced pressure. The crude intermediate was then dissolved in 4 mL of DMF and Fmoc-D-Asp(t-Bu)-OH (388 mg, 0.9438 mmol, 3 eq.), HATU (359 mg, 0.9438 mmol, 3 eq.), DIPEA (219 μL, 1.2584 mmol, 4 eq.) were added and stirred at room temperature for 24 h. The solvent was then evaporated and the mixture was stirred in 20% piperidine in DMF for 30 min. The reaction mixture was then diluted with EtOAc and washed with water (3x) and brine. The combined organics were washed with anhydrous Na 2 SO 4 Dry, filter, concentrate, and collect on CH 2 Cl 2 CH2 from 1% to 10% 3 Purification was performed by flash chromatography on silica gel eluting with 0.1% OH. 138 mg of product 7 was obtained in 61% yield. ESI-MS: m / z was C 36 H 50 N 7 O 9 [M+H] + Calculated for 724.8; found 724.4.

[0350] Compound JHU1221785: To a stirred solution of 7 (25 mg, 34.54 μmol, 1.0 eq.), 6-fluoronicitinic acid (14.6 mg, 103.62 μmol, 3 eq.) and HATU (39.4 mg, 103.62 μmol, 3 eq.) in DMF (1 mL), DIPEA (24 μL, 135.15 μmol, 4 eq.) was added and the mixture was allowed to stir at room temperature overnight. The reaction mixture was then diluted with EtOAc and washed with water (3x) and brine. The combined organics were washed with anhydrous Na 2 SO 4 The mixture was dried over 100 ml of TFA / CH 2 Cl 2 (1:1) was added and stirred for 30 min. The mixture was then concentrated and loaded onto a 12 g C18 cartridge (Biotage Sfar). The product was purified using a MeCN / water / TFA gradient (5 / 100 / 0.1 to 50 / 10 / 0.1). After lyophilization, 10 mg of JHU1221785 was obtained as a white solid in 40% yield. NMR (400 MHz, CD 3 OD): δ9.00-8.95(m, 1H), 8.62(d, J=13.2Hz, 1H), 8.39-8.26(m, 1H), 8.18-8.13(m, 1H), 8.13-8.0 8(m, 1H), 7.98-7.92(m, 1H), 7.79-7.72(m, 1H), 7.15-7.08(m, 1H), 4.77-4.83(m, 1H), 4.72-4.68( m, 1H), 4.40(s, 2H), 4.37-4.26(comp, 2H), 3.86-3.77(m, 1H), 3.70-3.60(m, 1H), 3.50-3.41(com p, 2H), 2.98-2.66(comp, 5H), 2.37-2.26(comp, 2H), 2.26-2.17(comp, 2H), 2.15-2.05(comp, 2H). ESI-MS: m / z is C 34 H 36 FN 8 O 10 [M+H] + Calculated 735.7; found 735.3.

[0351] [ka] Scheme 3. 18 Synthesis of F-JHU1221785.

[0352] Methyl (6-hydroxyquinoline-4-carbonyl)glycinate (8): 6-hydroxyquinoline-4-carboxylic acid (1) (200 mg, 1.05 mmol, 1.0 eq.), glycine methyl HCl salt (200 mg, 1.58 mmol, 1.5 eq.) and HATU (603 mg, 1.58 mmol, 1.5 eq.) were dissolved in 5 mL anhydrous DMF. DIPEA (0.46 mL, 2.64 mmol, 2.5 eq.) was added to the solution. The reaction was stirred at room temperature for 6 h. After removing the solvent under reduced pressure, the mixture was loaded onto a 25 g C18 cartridge and the product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 2 was obtained as a yellow powder with a yield of 76% (209 mg). 1 H-NMR (400MHz, CD 3 OD): δ8.69(s, 1H), 7.94(d, J=7.92Hz, 1H), 7.57-7.51(m, 3H), 7.42-7.37(m, 1H), 4.21(s, 2H), 3.81(s, 3H). 13 C-NMR (100 MHz, CD 3 OD): δ172.4, 160.9, 145.1, 143.7, 129.7, 129.4, 128.3, 121.8, 119.6, 112.4, 109.1, 56.8, 44.8.ESI-MS: m / z is C 13 H 13 N 2 O 4 [M+H] + Calculated for 261.3; findings 261.0.

[0353] Methyl (6-(3-((tert-butoxycarbonyl)amino)propoxy)quinoline-4-carbonyl)glycinate (9): Methyl (6-hydroxyquinoline-4-carbonyl)glycinate (8) (100 mg, 0.38 mmol, 1.0 eq.) and 3-(Bocamino)propyl bromide (230 mg, 0.96 mmol, 2.5 eq.) were dissolved in 4 mL of anhydrous DMF. 2 CO 3 (376 mg, 1.15 mmol, 3.0 eq.) was added to the solution and the reaction was stirred at room temperature overnight. After filtration, the solvent was removed under reduced pressure and the remaining mixture was loaded onto a 25 g C18 cartridge. The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 3 was obtained as a yellow powder with a yield of 54% (86 mg). 1 H-NMR (400 MHz, CDCl 3 ):δ8.68-8.37(m, 2H), 8.02(d, J=9.1Hz, 1H), 7.80(s, 1H), 7.72-7.64(m, 1H), 7.40(d, J=9.1Hz, 1H), 4.94(br s, 1H), 4.41-4.31(m, 2H), 4.27-4.18(m, 2H), 3.85(s, 3H), 3.44-3.30(m, 2H), 2.13-2.00(m, 2H), 1.43(s, 9H). 13 C NMR (100 MHz, CDCl 3 ):δ170.1, 167.2, 158.4, 144.7, 142.3, 128.4, 126.1, 124.7, 119.1, 103.7, 79.5, 60.4, 52.5, 41.4, 37.7, 29.3, 28.4.ESI-MS: m / z is C 21 H 28 N 3 O 6 [M+H] + Calculated for 418.5; found 418.1.

[0354] tert-Butyl (S)-(3-((4-((2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)carbamate (11): Compound 9 (145 mg, 0.347 mmol, 1.0 eq.) and LiOH (58.4 mg, 1.39 mmol, 4.0 eq.) were dissolved in 5 mL of H 2 The mixture was stirred in 200 ml of 1:1 hexane / THF for 6 h. After most of the THF was removed under reduced pressure, the mixture was loaded onto a 25 g C18 cartridge and eluted with a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1) to remove salts. After removal of the solvent and drying, the product 10 was obtained in quantitative yield. To a stirred solution of 10 (144 mg, 0.357 mmol, 1.0 eq.) in anhydrous DMF (4 mL) was added (S)-pyrrolidine-2-carbonitrile (56.8 mg, 0.428 mmol, 1.2 eq.), HATU (163 mg, 0.428 mmol, 1.2 eq.) and DIPEA (125 μL, 0.714 mmol, 2.0 eq.). After 6 h, the solvent was removed under reduced pressure and the crude mixture was loaded onto a 25 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 11 was obtained in 80% (135 mg) yield. 1 H NMR (400 MHz, CDCl 3 ):δ8.73(s, 1H), 7.95(d, J=10.2Hz, 1H), 7.68(br s, 1H), 7.63-7.56(m, 1H), 7.56-7.48(m, 1H), 7.38-7.29(m, 1H), 5.27(br s, 1H), 4.84-4.72(m, 1H), 4.46-4.35(m, 1H), 4.33-4.20(m, 1H), 4.17-4.09(m, 2H), 3.78-3.6 4(m, 1H), 3.59-3.46(m, 1H), 3.36(s, 2H), 2.38-2.17(m, 4H), 1.42(s, 9H), 1.35-1.27(m, 2H). 13 C NMR (100 MHz, CDCl 3):δ167.6, 167.5, 157.9, 156.2, 146.3, 130.2, 125.7, 123.7, 119.3, 118.0, 10 3.3, 79.0, 65.9, 46.8, 45.7, 42.2, 37.6, 29.8, 29.3, 28.4, 25.1.ESI-MS: m / z is C 25 H 32 N 5 O 5 [M+H] + Calculated for 482.6; findings 482.2.

[0355] (S)-6-(3-aminopropoxy)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)quinoline-4-carboxamide (4): Compound 11 (100 mg, 0.207 mmol.) was treated with 1 mL of a solution of TFA / methylene chloride (1 / 1) for 2 h. The solvent was removed under reduced pressure and the crude mixture was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, product 6 was obtained in quantitative yield. 1 H NMR (500 MHz, D 2 O): δ9.05(dd, J=2.0, 5.5Hz, 1H), 8.24(dd, J=2.0, 9.0Hz, 1H), 8.13(dd, J=2.0, 5.5Hz, 1H), 7.93-7.80(m, 2H), 4.84(t, J=4.5Hz, 2 H), 4.58-4.32(m, 4H), 3.87-3.78(m, 1H), 3.67(q, J=8.0Hz, 1H), 3.29(t, J=6.0Hz, 2H), 2.45-2.12(m, 6H), 1.34(d, J=6.0Hz, 1H). ESI-MS: m / z is C 20 H 23 N 5 O 3 [M+H] + Calculated 382.4; found 382.2.

[0356] [ka] Scheme 4. Synthesis of FAP-acetylene 1.

[0357] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-6-(3-(pent-4-ynamido)propoxy)quinoline-4-carboxamide (FAP-acetylene 1): To a stirred solution of 4 (10 mg, 0.026 mmol, 1.0 eq.) in DMF (1 mL) was added 2,5-dioxopyrrolidin-1-ylpent-4-ynoate (12) (5.12 mg, 0.026 mmol, 1.0 eq.) and DIPEA (13.5 μL, 0.078 mmol, 3.0 eq.). After 2 h, the solvent was removed under reduced pressure and the crude mixture was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (from 0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, 8.6 mg of FAP-acetylene 1 was obtained in 71% yield. 1 H NMR (500MHz, DMSO-d6): δ9.05 (t, J=5.5Hz, 1H), 8.84 (d, J=4.5Hz, 1H), 8.03-7.95 (m, 2H), 7. 86(d, J=2.0Hz, 1H), 7.56(d, J=4.0Hz, 1H), 7.49(dd, J=2.5, 9.0Hz, 1H), 4.85-4.78(m, 1H), 4. 17(t, J=6.0Hz, 2H), 2.92-2.86(m, 2H), 2.81(s, 1H), 2.74(t, J=2.5Hz, 1H), 2.37-2.31(m, 3H) ), 2.29-2.23(m, 3H), 2.22-2.13(m, 2H), 2.12-1.99(m, 3H), 1.97-1.89(m, 2H); ESI-MS: m / z is C 25 H 28 N 5 O 4 [M+H] + Calculated 462.2; found 462.1.

[0358] [ka] Reagents and conditions: a) (i). 2,5-Dioxopyrrolidin-1-yl 5-((tert-butoxycarbonyl)amino)-pentanoate, DIPEA, DMF, RT, overnight. (ii). TFA:CH 2 Cl 2 (1:1), RT, 2h, 75%; (b) (i) Fmoc-L-Lys(Boc)-OSu, DIPEA, DMF, RT, 2h; (ii) 20% piperidine, DMF, RT, 1h, 65%; (c) Azide-PEG3-NHS ester, DIPEA, DMSO, RT, 2h, 70%; (d) FAP-acetylene 1, CuSO 4 .5H 2 O, sodium ascorbate, tBuOH / DMSO / H 2 HO, RT, overnight, 86%; e) (i).TFA:CH 2 Cl 2 (1:1), RT, 2h, 90%; (ii).DOTA-NHS-ester, DIPEA, DMSO, RT, 2h, 69%. Scheme 5. Synthesis of SB-FAP-01

[0359] (((S)-5-(5-amino-N-(4-bromobenzyl)pentanamido)-1-carboxypentyl)carbamoyl)-L-glutamic acid (14): To a stirred solution of 13 (214 mg, 0.326 mmol, 1.0 eq.) and 2,5-dioxopyrrolidin-1-yl 5-((tert-butoxycarbonyl)amino)pentanoate (102.5 mg, 0.326 mmol, 1.0 eq.) in DMF (2 mL) was added DIPEA (170 μL, 0.978 mmol, 3.0 eq.) at room temperature. The reaction mixture was stirred for 24 h and concentrated to give the crude product. The crude was diluted with 2 mL of TFA / CH 2 Cl 2 (1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 14 (143 mg, 75%) was given as a white solid. H 1-NMR (500MHz, D 2 O): δ7.55(d, J=8.5Hz, 1H), 7.51(d, J=8.5Hz, 1H), 7.12(dd, J=2.5, 8.0Hz, 2H), 4.59(s, 1H), 4.51(s, 1H), 4.29-4.22(m, 1H), 4.19-4.11(m, 1H), 3.39-3.29(m, 2H), 3.05-2.88(m, 2H), 2.59-2.38(m, 4H), 2.22-2 .10(m, 1H), 2.02-1.90(m, 1H), 1.82-1.46(m, 8H), 1.39-1.24(m, 2H).

[0360] (10S,23S,27S)-10-amino-18-(4-bromobenzyl)-2,2-dimethyl-4,11,17,25-tetraoxo-3-oxa-5,12,18,24,26-pentaazanonacosane-23,27,29-tricarboxylic acid (15): To a stirred solution of 14 (139 mg, 0.236 mmol, 1.0 eq.) and Fmoc-L-Lys(Boc)-OSu (133.9 mg, 0.236 mmol, 1.0 eq.) in DMF (2 mL) was added DIPEA (250 μL, 1.42 mmol, 6.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product. To the above crude was added 2 mL of 20% piperidine in DMF (3 mL) at room temperature and the mixture was stirred for 1 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 15 (125 mg, 65%) was given as a white solid. 1 -NMR (500MHz, DMSO-d 6):δ8.43-8.32(m, 1H), 8.08(s, 2H), 7.93(d, J=7.5Hz, 1H), 7.74(d, J=7.5Hz, 1H), 7.55( d, J=8.0Hz, 1H), 7.41(d, J=8.0Hz, 1H), 6.74(t, J=5.0Hz, 1H), 6.40-6.26(m, 2H), 4.52(s , 1H), 4.44(s, 1H), 4.14-3.98(m, 2H), 3.23-2.97(m, 5H), 2.93-2.82(m, 2H), 2.37(t, J=7 [M+H] + is C 35 H 56 BrN 6 O 11 Calculated for, 815.3192; findings, 815.3184.

[0361] (11S,24S,28S)-1-Azido-19-(4-bromobenzyl)-11-(4-((tert-butoxycarbonyl)amino)butyl)-9,12,18,26-tetraoxo-3,6-dioxa-10,13,19,25,27-pentaazatriacontane-24,28,30-tricarboxylic acid (16): To a stirred solution of 15 (47.3 mg, 0.058 mmol, 1.0 eq.) and azido-PEG3-NHS ester (20 mg, 0.058 mmol, 1.0 eq.) in DMSO (100 μL) was added DIPEA (80 μL, 0.464 mmol, 8.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 11 (42 mg, 70%) was given as a white solid. ESI-MS: m / z was C 44 H 71 BrN 9 O 15 [M+H] + Calculated 1044.4; findings 1044.3.

[0362] Compound 17: A stirred solution of FAP-acetylene 1 (6 mg, 0.0129 mmol, 1.0 eq.) in t-BuOH:HO (1:2, 300 μL) was added with CuSO 4 .5H 2 HO (0.486 mg, 0.0019 mmol, 0.15 eq.) and sodium ascorbate (1.15 mg, 0.0058 mmol, 0.45 eq.) were added. To the above reaction mixture, a solution of product 16 (13.5 mg, 0.0129 mmol, 1.0 eq.) in DMSO (100 μL) was added and stirred overnight at room temperature. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 17 (16.7 mg, 86%) was given as a white solid. ESI-MS: m / z was C 69 H 98 BrN 14 O 19 [M+H] + Calculated 1505.6; findings 1505.4.

[0363] Compound SB-FAP-01: Compound 17 (9 mg, 0.0059 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, it gave the free amine in quantitative yield. To a stirred solution of the amine (3.8 mg, 0.0027 mmol, 1.0 eq.) and DOTA-NHS-ester (3.08 mg, 0.004 mmol, 1.5 eq.) in DMSO (50 μL) was added DIPEA (4.7 μL, 0.027 mmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude was purified by preparative RP-HPLC chromatography using 0.1% TFA in HO and 0.1% TFA in acetonitrile as eluents followed by lyophilization to give compound SB-FAP-01 (3.3 mg, 69%) as a white solid. RP-HPLC purification was achieved using an Agilent System, λ254 nm, 250 mmX10 mm Phenomenex Luna C18 column, solvent gradient: 90% HO (0.1% TFA) and 10% ACN (0.1% TFA), reaching 90% ACN in 25 minutes at 5 mL / min flow rate, product eluted at 13.8 minutes]. HRMS (ESI) m / z: [M+H] + is C 80 H 114 BrN 18 O 24 Calculated for, 1789.7456; found for, 1789.7442.

[0364] [ka] Scheme 6. Synthesis of SB-FAP-02.

[0365] Compound SB-FAP-02: Compound 17 (8 mg, 0.0053 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, it gave the free amine in quantitative yield. To a stirred solution of the amine (6.7 mg, 0.000.0047 mmol, 1.0 eq.) and NOTA-NHS-ester (3.46 mg, 0.0052 mmol, 1.1 eq.) in DMSO (50 μL) was added triethylamine (6.6 μL, 0.047 mmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude product was purified using H 2 Purification was achieved by preparative RP-HPLC chromatography using 0.1% TFA in 0 and acetonitrile in 0.1% TFA as eluents followed by lyophilization to give compound SB-FAP-02 (5.8 mg, 72%) as a white solid. [RP-HPLC purification was achieved using an Agilent System, λ254 nm, 250 mm×10 mm Phenomenex Luna C18 column, solvent gradient: 98% HO (0.1% TFA) and 2% ACN (0.1% TFA), flow rate of 5 mL / min reaching 50% ACN from 0 to 20 min, 90% ACN from 20 to 25 min, product eluted at 16.8 min]. 1H-NMR (500MHz, DMSO-d6): δ12.11 (Broad singlet, 5H), 9.04(t, J=5.5Hz, 1H), 8.84-8.80(m, 1H), 8.19-8.10(m, 1H), 8.03-7.90(m, 3H), 7.59-7.43(m, 4H) , 7.19-7.10(m, 2H), 6.37-6.25(m, 2H), 4.82(s, 2H), 4.51(m, 2H), 4.43(s, 2H), 4.26-4.02(m, 14H), 3.79-3.66(m , 6H), 3.64-3.56(m, 6H), 3.44(d, J=16.5, 8H), 3.29-2.94(m, 11H), 2.86-2.79(m, 2H), 2.42(t, J=7.0, 2H), 2.39- 2.30(m, 3H), 2.28-2.16(m, 4H), 2.12-2.01(m, 2H), 1.96-1.86(m, 2H), 1.73-1.17(m, 20H);HRMS(ESI)m / z:[M+H] + is C 76 H 109 BrN 17 O 22 Calculated for, 1690.7129; Found for, 1690.7110.

[0366] [ka] Scheme 7. Synthesis of SB-FAP-03.

[0367] Compound SB-FAP-03: Compound 17 (8 mg, 0.0053 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2 (1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, the free amine (6.7 mg) was obtained in quantitative yield. The mixture of the amine (6.0 mg, 0.0042 mmol, 1.0 eq.) and DOTAGA ( t Bu)4 To a stirred solution of the -NHS-ester (4.08 mg, 0.0064 mmol, 1.2 eq.) was added diisopropylethylamine (5.9 μL, 0.042 mmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude was purified by preparative RP-HPLC chromatography to give the product (5.4 mg, 61%) as a white solid. The stirred solution of the above product was added to TFA / H 2 A cocktail of O / TIPS (950 μL / 25 μL / 25 μL) was added at room temperature. The reaction mixture was stirred for 24 h, and the crude product was purified by H 2 Purified by preparative RP-HPLC chromatography using 0.1% TFA in O and acetonitrile in 0.1% TFA as eluents followed by lyophilization to give compound SB-FAP-03 (2.7 mg, 56%) as a white solid. HRMS (ESI) m / z: [M+H] + is C 83 H 120 BrN 18 O 26 Calculated for, 1863.7812; findings, 1863.7799.

[0368] [ka] Scheme 8. Synthesis of SB-FAP-04.

[0369] Compound SB-FAP-04: Compound 17 (8 mg, 0.0053 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, the free amine was obtained in quantitative yield. To a stirred solution of the amine (1.9 mg, 0.00137 mmol, 1.5 eq.) and IR800CW-NHS ester (1.07 mg, 0.000917 mmol, 1.0 eq.) in DMSO (20 μL) was added DIPEA (2.0 μL, 0.0137 mmol, 15.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the crude was purified by preparative RP-HPLC chromatography using 20 mM triethylammoniumacetate buffer and acetonitrile as eluent followed by lyophilization to give compound SB-FAP-04 (1.6 mg, 77%) as a green solid. HRMS (ESI) m / z: [M / 2] + is C 110 H 143 BrN 16 O 31 S 4 Calculated for: 1195.409171; findings: 1195.4080.

[0370] [ka] Scheme 9. Synthesis of SB-FAP-05.

[0371] Compound SB-FAP-05: Compound 17 (8 mg, 0.0053 mmol) was dissolved in 1 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, the free amine was obtained in quantitative yield. To a stirred solution of the amine (1.43 mg, 1.01 μmol, 2.0 eq.) and IR800CW-NHS ester (1.0 mg, 0.51 μmol, 1.0 eq.) in DMSO (20 μL) was added DIPEA (2.7 μL, 15.3 mol, 30.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the crude was purified by preparative RP-HPLC chromatography using 20 mM triethylammonium acetate buffer and acetonitrile as eluent followed by lyophilization to give compound SB-FAP-05 (0.95 mg, 59%) as a green solid.

[0372] [ka] Scheme 10. Synthesis of SB-FAP-06, SB-FAP-07, and SB-FAP-08.

[0373] 2,5-dioxopyrrolidin-1-yl(S)-2-((tert-butoxycarbonyl)amino)-6-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)hexanoate (18):DMSO:H 2 A stirred solution of FAP-acetylene 1 (10 mg, 0.0216 mmol, 1.0 eq.) and Boc-Lys(N)-OH in 2HO (1:1, 400 µL) was added to a freshly prepared solution of CuSO. 4 .5H 2O (0.81 mg, 3.24 μmol, 0.15 eq.) and sodium ascorbate (1.93 mg, 9.74 μmol, 0.45 eq.) were added. The reaction mixture was stirred overnight at room temperature and concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, the triazole product (11.3 mg, 71%) was given as a white solid. To a stirred solution of the triazole product (8.5 mg, 0.0115 mmol, 1.0 eq.) in dimethylformamide (500 μL) was added TSTU (5.23 mg, 0.0173 mmol, 1.5 eq.) and diisopropylethylamine (6.0 μL, 0.034 mmol, 3.0 eq.) at room temperature under nitrogen atmosphere. The reaction mixture was stirred for 2 h and concentrated to give the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 18 (6.0 mg, 63%) was obtained as a white solid.

[0374] (((S)-5-(6-amino-N-(4-bromobenzyl)hexanamido)-1-carboxypentyl)carbamoyl)-L-glutamic acid (19): To a stirred solution of 13 (365 mg, 0.556 mmol, 1.0 eq.) and 2,5-dioxopyrrolidin-1-yl 6-((tert-butoxycarbonyl)amino)hexanoate (174.7 mg, 0.556 mmol, 1.0 eq.) in DMF (2 mL) was added DIPEA (233 μL, 1.669 mmol, 3.0 eq.) at room temperature. The reaction mixture was stirred for 24 h and concentrated to give the crude product. The crude was diluted with 2 mL of TFA / CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 19 (360 mg, 98%) was given as a white solid.

[0375] Compound 20: To a stirred solution of 18 (6.9 mg, 8.3 μmol, 1.0 eq.) and 19 (7.47 mg, 12.4 μmol, 1.5 eq.) in DMF was added DIPEA (13.7 μL, 99.6 μmol, 12.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product. The crude was diluted with 1 mL of TFA:CH 2 Cl 2 (1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 15 (7.0 mg, 98%) was given as a white solid.

[0376] Compound SB-FAP-06: To a stirred solution of amine 20 (3.81 mg, 3.13 μmol, 1.0 eq.) and DOTA-NHS-ester (2.62 mg, 3.44 μmol, 1.1 eq.) in DMSO (50 μL) was added DIPEA (5.2 μL, 37.6 μmol, 12.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude product was purified by H 2 The mixture was purified by preparative RP-HPLC chromatography using 0.1% TFA in O and acetonitrile as eluents in 0.1% TFA followed by lyophilization to give compound SB-FAP-06 (4.0 mg, 81%) as a white solid.

[0377] Compound SB-FAP-07: To a stirred solution of amine 20 (3.4 mg, 2.79 μmol, 1.0 eq.) and NOTA-NHS-ester (2.2 mg, 3.35 μmol, 1.2 eq.) in DMSO (65 μL) was added DIPEA (4.7 μL, 33.5 μmol, 12.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude product was purified by H 2 The mixture was purified by preparative RP-HPLC chromatography using 0.1% TFA in O and acetonitrile as eluents in 0.1% TFA followed by lyophilization to give compound SB-FAP-06 (3.1 mg, 74%) as a white solid.

[0378] Compound SB-FAP-08: To a stirred solution of amine 20 (1.35 mg, 1.11 μmol, 1.3 eq.) and IR800CW-NHS-ester (1.0 mg, 0.85 μmol, 1.0 eq.) in DMSO (25 μL) was added DIPEA (1.8 μL, 12.86 μmol, 15.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude was purified by preparative RP-HPLC chromatography using 20 mM triethylammonium acetate buffer as eluent in acetonitrile followed by lyophilization to give compound SB-FAP-08 (0.85 mg, 78%) as a green solid.

[0379] [ka]

[0380] Compound 21: To a stirred solution of 18 (6.0 mg, 7.22 μmol, 1.0 eq.) and 4 (2.75 mg, 7.22 μmol, 1.0 eq.) in DMF (300 μL) was added DIPEA (7.5 μL, 943.3 μmol, 6.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product. The crude was diluted with 1 mL of TFA:CH 2 Cl 2(1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 21 (4.3 mg, 60%) was given as a white solid.

[0381] Compound SB-FAP-09: To a stirred solution of amine 21 (3.12 mg, 3.12 μmol, 1.0 eq.) and NOTA-NHS-ester (2.3 mg, 3.44 μmol, 1.1 eq.) in DMSO (75 μL) was added DIPEA (4.35 μL, 31.2 μmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and the crude product was purified by H 2 The mixture was purified by preparative RP-HPLC chromatography using 0.1% TFA in O and acetonitrile as eluents in 0.1% TFA followed by lyophilization to give compound SB-FAP-09 (2.8 mg, 70%) as a white solid.

[0382] [ka]

[0383] 2,2',2''-(10-(4-((2-((S)-2-amino-6-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)hexanamido)ethyl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (22): 13 (10.8 mg, 12.99 μmol, 1.0 eq.) and DOTAGA-NH 2To a stirred solution of (6.74 mg, 12.9 μmol, 1.0 eq.) was added triethylamine (21.7 μL, 156.0 μmol, 12.0 eq.) at room temperature. The reaction mixture was stirred for 2 h and concentrated to give the crude product. The crude was added to 1 mL of TFA:CH 2 Cl 2 (1:1) was added at room temperature and the mixture was stirred for 2 h. The reaction mixture was concentrated to obtain the crude product, which was loaded onto a 12 g C18 cartridge (Silicycle, Canada). The product was purified using a MeCN / water / TFA gradient (0 / 100 / 0.1 to 90 / 10 / 0.1). After lyophilization, compound 22 (4.3 mg, 43%) was given as a white solid.

[0384] 1-(6-(((2S)-1-((2-(4-carboxy-4-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl)butanamido)ethyl)amino)-6-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)-1-oxohexan(hexan)-2-yl)amino)-6-oxohexyl)-2-((E)-2-((E)-3-(2-((E)-3,3-dimethyl-5 -sulfo-1-(4-sulfobutyl)indolin-2-ylidene)ethylidene)-2-(4-sulfophenoxy)cyclohex-1-en-1-yl)vinyl)-3,3-dimethyl-3H-indol-1-ium-5-sulfonate (SB-FAP-10): To a stirred solution of amine 22 (1.63 mg, 1.44 μmol, 1.2 eq.) and IR800CW-NHS-ester (1.4 mg, 1.20 μmol, 1.0 eq.) in DMSO (25 μL) was added triethylamine (3.4 μL, 24.0 μmol, 20.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the crude was purified by preparative RP-HPLC chromatography using 20 mM triethylammonium acetate buffer as eluent in acetonitrile followed by lyophilization to give compound SB-FAP-08 (0.92 mg, 78%) as a green solid.

[0385] 2,2',2''-(10-(1-carboxy-4-((2-((S)-6-(4-(3-((3-((4-((2-((S)-2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)amino)-3-oxopropyl)-1H-1,2,3-triazol-1-yl)-2-(6-fluoronicotinamidohexanamido)ethyl)amino)-4-oxobutyl)-1 ,4,7,10-Tetraazacyclododecane-1,4,7-triyl)triacetic acid (SB-FAP-11): To a stirred solution of amine 22 (5.9 mg, 5.20 μmol, 1.0 eq.) and 2,5-dioxopyrrolidin-1-yl 6-fluoronicotinate (1.5 mg, 6.20 μmol, 1.2 eq.) in DMSO (50 μL) was added triethylamine (7.2 μL, 52.0 μmol, 10.0 eq.) at room temperature. The reaction mixture was stirred for 2 h, and the crude product was purified by H 2 The mixture was purified by preparative RP-HPLC chromatography using 0.1% TFA in O and acetonitrile as eluents in 0.1% TFA followed by lyophilization to give compound SB-FAP-11 (2.8 mg, 70%) as a white solid.

[0386] [ka]

[0387] Example 6 Representative FAP-α inhibitors

[0388] Representative FAP-α inhibitors and representative biological data are provided in Table 3.

[0389] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8]

[0390] Example 7 Representative FAP-α / PSMA dual inhibitor

[0391] Representative FAP-α / PSMA dual inhibitors and representative biological data are provided in Table 4.

[0392] [Table 4-1] [Table 4-2]

[0393] Example 8 Distribution in the body 111 In-SRI-08-15 and related compounds

[0394] Tissue biodistribution 111 In-SRI-8-15 and related compounds are provided in Table 5 (n=4, time point 1 h). Table 5, 111 As provided in In-SRI-08-15, based on the placement of the metal chelator on the DOTA ring, extremely high uptake is unexpectedly possible in U87 cells. 18 Unlike F]FAPI-74, Al[ 18 F]F chemistry is 18It is not used to introduce F. In contrast, the compounds of the present disclosure are prepared in one step from trimethylammonium precursors with extremely high specific activity. [ka]

[0395] [Table 5]

[0396] Example 9 Representative compounds for biodistribution [ka]

[0397] Head-to-head biodistribution study (%ID / g) (n=5 / group). Dose 20μCi. 1h time point 64 Cu-Sri-07-86 and 64 Cu-FAPI-74. 64 Cu-FAPI-04 contains a DOTA group and still exhibits low liver uptake; it is noted that the hepatic absorption rate is low, most likely related to the piperazine group next to the DOTA, acting as a co-ligand. The DOTA chelate showed a liver uptake of >10% ID / g.

[0398] [Table 6]

[0399] [Table 7]

[0400] [ka]

[0401] Compound name: 64Cu-Sri-06-98. Dose: 200 μCi, n=3 per group. Location of the triazole group and liver uptake are noted.

[0402] [Table 8]

[0403] [Table 9]

[0404] [ka]

[0405] Compound name: 64 Cu-SRI-06-57, 64 Cu-FAPI-04, 64 Cu-07-56 and 64 Cu-07-05 Dose: 20 μCi, n=3 per group, tumor model: U87 in NSG mice. Note the location of the triazole group and DOTA.

[0406] [Table 10]

[0407] [Table 11]

[0408] [ka]

[0409] Dose: 20 μCi, n=3 at 1 h and n=4 at 2 and 4 h per group, tumor model: SK-MEL-24 in NSG mice

[0410] [Table 12]

[0411] [Table 13] Tumor Weight:

[0412] [ka]

[0413] In male NSG mice bearing U87 xenografts in the upper flank at 2 h 64 Cu-Sri-06-57 (SB-FAP-2) and 64 Tissue biodistribution data for Cu-FAPI-04 (n=3-4) (data are %ID / g and expressed as mean ± SD).

[0414] [Table 14]

[0415] [Table 15]

[0416] [Table 16]

[0417] [ka]

[0418] 55 Biodistribution study (n=4) using Co-SB-FAP-01(DOTA)AT2h in NCA.MB-MDA-231NSG mice (U87 tumors were not available), dose: ~10 μCi.

[0419] [Table 17]

[0420] [ka]

[0421] [Table 18]

[0422] Example 10 Synthesis of FAP-triamine compounds [ka]

[0423] Example 11 Introduction of DOTA chelator of lysine [ka]

[0424] Example 12 Sc- 18 F labeled FAPI The one below is Sc- 18 1 is a scheme for F-labeled FAPI and representative compounds. [ka]

[0425] Example 13 Biodistribution of representative compounds

[0426] [Table 19]

[0427] [Table 20]

[0428]

Table 21

[0429]

Table 22

[0430]

Table 23

[0431]

Table 24

[0432]

Table 25

[0433]

Table 26

[0434]

Table 27

[0435]

Table 28

[0436]

Table 29

[0437]

Table 30

[0438]

Table 31

[0439]

Table 32

[0440]

Table 33

[0441]

Table 34

[0442]

Table 35

[0443]

Table 36

[0444]

Table 37

[0445]

Table 38

[0446]

Table 39

[0447]

Table 40

[0448]

Table 41

[0449] [Table 42]

[0450] [Table 43]

[0451] [Table 44]

[0452] [Table 45]

[0453] References

[0454] All publications, patent applications, patents, and other references mentioned in this specification are indicative of the level of skill of those in the art to which the subject matter of this disclosure pertains. All publications, patent applications, patents, and other references are incorporated herein by reference to the same extent as if each individual publication, patent application, patent, and other reference was specifically and individually indicated to be incorporated by reference. Although several patent applications, patents, and other references are referenced herein, it will be understood that such reference is not an admission that any of these documents form part of the common general knowledge in the art. In the event of a conflict between this specification and an incorporated reference, this specification (including any amendments thereof that may be based on the incorporated reference) shall control. Standard art-recognized meanings of terms are used herein unless otherwise indicated. Standard abbreviations for various terms are used herein.

[0455] Allinen M, Beroukhim R, Cai L, Brennan C, Lahti-Domenici J, Huang H, Porter D, Hu M, Chin L, Richardson A, Schnitt S, Sellers WR, Polyak K. Molecular characterization of the tumor microenvironment in breast cancer. Cancer Cell. 2004 Jul;6(1):17-32. Bae S, Park CW, Son HK, Ju HK, Paik D, Jeon CJ, Koh GY, Kim J, Kim H. Fibroblast activation protein alpha identifies mesenchymal stromal cells from human bone marrow. Br J Haematol.(British Journal of Haematology)2008 Sep;142(5):827-30. Bandari RP, Carmack TL, Malhotra A, Watkinson L, Fergason Cantrell EA, Lewis MR, et al., Development of heterobivalent theranostic probes having high affinity / selectivity for the GRPR / PSMA. Journal of Medicinal Chemistry. 2021;64:2151-66. Banerjee et al., Preclinical Evaluation of 203 / 212Pb-Labeled Low-Molecular-Weight Compounds for Targeted Radiopharmaceutical Therapy of Prostate Cancer, J. Nucl. Med, 61 (1), 80-88 (2020). Banerjee SR, Kumar V, Lisok A, Chen J, Minn I, Brummet M et al. 177Lu-labeled low-molecular-weight agents for PSMA-targeted radiopharmaceutical therapy. Eur J Nucl Med Mol Imaging. 2019;46:2545-57. Banerjee SR, Pullambhatla M, Byun Y, Nimmagadda S, Foss CA, Green G et al., Sequential SPECT and Optical Imaging of Experimental Models of Prostate Cancer with a Dual Modality Inhibitor of the Prostate-Specific Membrane Antigen. Angewandte Chemie International Edition. 2011;50:9167-70. Bejarano L, Jordão MJC, Joyce JA. Therapeutic targeting of the tumor microenvironment. Cancer Discovery. 2021;11:933-59. Brennen WN, Isaacs JT, Denmeade SR. Rationale behind targeting fibroblast activation protein-expressing carcinoma-associated fibroblasts as a novel chemotherapeutic strategy. Molecular Cancer Therapeutics. 2012;11:257-66. Busek P, Balaziova E, Matrasova I, Hilser M, Tomas R, Syrucek M et al. Fibroblast activation protein alpha is expressed by transformed and stromal cells and is associated with mesenchymal features in glioblastoma. Tumor Biology. 2016;37:13961-71. Chang SS, Reuter VE, Heston WDW, Bander NH, Grauer LS. Five different anti-prostate-specific membrane antigen (PSMA) antibodies confirm PSMA expression in tumor-associated neovasculature. Cancer Res. 1999;59:3192. Chen ZY, Wang YX, Lin Y, Zhang JS, Yang F, Zhou QL, Liao YY. Advance of molecular imaging technology and targeted imaging agent in imaging and therapy. Biomed Res Int. 2014;2014 :819324. PMCID:PMC3943245. Cheng Y, Prusoff WH. Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol. 1973;22:3099-108. Cho et al., Biodistribution, tumor detection, and radiation dosimetry of 18F-DCFBC, a low-molecular-weight inhibitor of prostate-specific membrane antigen, in patients with metastatic prostate cancer. Journal of nuclear medicine : official publication, Society of Nuclear Medicine. 2012;53:1883-1891. Cho, SY et al. Biodistribution, tumor detection, and radiation dosimetry of 18F-DCFBC, a low-molecular-weight inhibitor of prostate-specific membrane antigen, in patients with metastatic official prostate cancer. Journal of nuclear medicine : publication, Society of Nuclear Medicine. 2012;53:1883-1891. Coenen HH, Elsinga PH, Iwata R, Kilbourn MR, Pillai MR, Rajan MG, Wagner HN Jr. Zaknun JJ. Fluorine-18 radiopharmaceuticals beyond [18F]FDG for use in oncology and neurosciences. Nuclear medicine and biology. 2010;37:727-740. Cohen A, Larson SM, Old LJ, et al. Antibody targeting in metastatic colon cancer: a phase I study of monoclonal antibody F19 against a cell-surface protein of reactive tumor stromal fibroblasts. J Clin Oncol. 1994 Jun;12(6):1193-203. Cohen SJ, Alpaugh RK, Palazzo I, Meropol NJ, Rogatko A, Xu Z, et al. Fibroblast activation protein and its relationship to clinical outcome in pancreatic adenocarcinoma. Pancreas. 2008;37:154-8. Conway RE, Joiner K, Patterson A, Bourgeois D, Rampp R, Hannah BC, et al. Prostate specific membrane antigen produces pro-angiogenic laminin peptides downstream of matrix metalloprotease-2. Angiogenesis. 2013;16:847-60. Delgado-Bellido D, Serrano-Saenz S, Fernandez-Cortes M, Oliver FJ. Vasculogenic mimicry signaling revisited: focus on non-vascular VE-cadherin. Molecular Cancer. 2017;16:65. Eder M, Schafer M, Bauder-Wust U, Haberkorn U, Eisenhut M, Kopka K. Preclinical evaluation of a bispecific low-molecular heterodimer targeting both PSMA and GRPR for improved PET imaging and therapy of prostate cancer. The Prostate. 2014;74:659-68. Edosada CY, Quan C, Tran T, Pham V, Wiesmann C, Fairbrother W, Wolf BB. Peptide substrate profiling defines fibroblast activation protein as an endopeptidase of strict Gly(2)-Pro(1)-cleaving specificity. FEBS Lett. 2006 Mar 6;580(6):1581-6. Errarte P, Guarch R, Pulido R, Blanco L, Nunes-Xavier CE, Beitia M, et al. The Expression of Fibroblast Activation Protein in Clear Cell Renal Cell Carcinomas Is Associated with Synchronous Lymph Node Metastases. PloS one. 2016;11:e0169105. Fischer E, Chaitanya K, Wuest T et al. Radioimmunotherapy of fibroblast activation protein positive tumors by rapidly internalizing antibodies. Clin Cancer Res. 2012;18:6208-6218. Fitzgerald AA, Weiner LM. The role of fibroblast activation protein in health and malignancy. Cancer and Metastasis Reviews. 2020;39:783-803. Franco OE, Shaw AK, Strand DW, Hayward SW. Cancer associated fibroblasts in cancer pathogenesis. Semin Cell Dev Biol. 2010 Feb;21(1):33-9. Garin-Chesa P, Old LJ, Rettig WJ. Cell surface glycoprotein of reactive stromal fibroblasts as a potential antibody target in human epithelial cancers. Proc Natl Acad Sci USA. 1990 Sep;87(18):7235-9. PMCID:PMC54718. He M, Henderson M, Muth S, Murphy A, Zheng L. Preclinical mouse models for immunotherapeutic and non-immunotherapeutic drug development for pancreatic ductal adenocarcinoma. Annals of Pancreatic Cancer. 2020;3. Herrmann K, Schwaiger M, Lewis JS, Solomon SB, McNeil BJ, Baumann M et al. Radiotheranostics: a roadmap for future development. The Lancet Oncology. 2020;21:e146-e56. Hintz HM, Gallant JP, Vander Griend DJ, Coleman IM, Nelson PS, LeBeau AM. Imaging fibroblast activation protein alpha improves diagnosis of metastatic prostate cancer with positron emission tomography. Clinical Cancer Research. 2020;26:4882-91. Hofman MS, Emmett L, Sandhu S, Iravani A, Joshua AM, Goh JC, et al., [177Lu]Lu-PSMA-617 versus cabazitaxel in patients with metastatic castration-resistant prostate cancer (TheraP): a randomised, open-label, phase 2 trial. The Lancet. 2021;397:797-804. Hofman MS, Lawrentschuk N, Francis RJ, Tang C, Vela I, Thomas P, et al. Prostate-specific membrane antigen PET-CT in patients with high-risk prostate cancer before curativeintent surgery or radiotherapy (proPSMA): a prospective, randomised, multicentre study. The Lancet. 2020;395:1208-16. Huang, X.;Bennett, M.;Thorpe, P. E. Prostate 2004, 61, 1. Imlimthan S, Moon ES, Rathke H, Afshar-Oromieh A, Rosch F, Rominger A et al. New frontiers in cancer imaging and therapy based on radiolabeled fibroblast activation protein inhibitors: A rational review and current progress. Pharmaceuticals. 2021;14:1023. International PCT Patent Application No. WO 2010 / 108125 A2, PSMA-Targeting Compounds and Uses Thereof, to Pomper et al., published Sept. 23, 2010. International PCT Patent Application No. WO2018232280, PSMA Targeted Fluorescent Agents for Image Guided Surgery, to Pomper et al., published Dec. 20, 2018. International PCT Patent Application No. WO2019 / 154886, FAP Inhibitor, Haberkorn et al., published August 15, 2019. International PCT Patent Application No. WO 2013 / 107820, for Novel FAP Inhibitors, to Jansen et al., issued July 25, 2013. Isik EG, Has-Simsek D, Sanli O, Sanli Y, Kuyumcu S. Fibroblast activation protein-targeted pet imaging of metastatic castration-resistant prostate cancer compared with 68Ga-PSMA and 18F-FDG PET / CT. Clinical Nuclear Medicine. 2021. Jambunathan K, Watson DS, Endsley AN, Kodukula K, Galande AK, Comparative analysis of the substrate preferences of two Post-proline cleaving endopeptidases, prolyl oligopeptidase and fibroblast activation protein α, FEBS Lett. 2012;586(16) :2507-2512, Jansen K, Heirbaut L, Cheng JD, Joossens J, Ryabtsova O, Cos P, Maes L, Lambeir AM, De Meester I, Augustyns K, Van der Veken P. Selective Inhibitors of Fibroblast Activation Protein (FAP) with a (4-Quinolinoyl)-glycyl-2-cyanopyrrolidine Scaffold. ACS Med Chem Lett. 2013 Mar 18;4(5):491-6. Jansen K, Heirbaut L, Verkerk R, Cheng JD, Joossens J, Cos P, et al. Extended structure-activity relationship and pharmacokinetic investigation of (4-Quinolinoyl)glycyl-2-cyanopyrrolidine Inhibitors of Fibroblast Activation Protein (FAP). Journal of Medicinal Chemistry. 2014;57:3053-74. Juillerat-Jeanneret L, Tafelmeyer P, Golshayan D. Fibroblast activation protein-alpha in fibrogenic disorders and cancer: more than a prolyl-specific peptidase? Expert Opin. Ther. Targets 2017;21:977-991. Kalluri R. The biology and function of fibroblasts in cancer. Nature Reviews Cancer. 2016;16:582-98. Keane FM, Yao TW, Seelk S, Gall MG, Chowdhury S, Poplawski SE et al., Quantitation of fibroblast activation protein (FAP)-specific protease activity in mouse, baboon and human fluids and organs. FEBS Open Bio. 2014;4:43-54. Kelly T. Fibroblast activation protein-alpha and dipeptidyl peptidase IV (CD26): cell-surface proteases that activate cell signaling and are potential targets for cancer therapy. Drug Resist Update. 2005 Feb-Apr;8(1-2):51-8. Kesch C, Yirga L, Dendl K, Handke A, Darr C, Krafft U, et al. High fibroblast-activation-protein expression in castration-resistant prostate cancer supports the use of FAPI-molecular theranostics. European Journal of Nuclear Medicine and Molecular Imaging. 2021;49:385-9. Kessel K, Seifert R, Weckesser M, Boegemann M, Huss S, Kratochwil C et al. Prostate-specific membrane antigen and fibroblast activation protein distribution in prostate cancer: preliminary data on immunohistochemistry and PET imaging. Annals of Nuclear Medicine. 2021. Kraman M, Bambrough PJ, Arnold JN, Roberts EW, Magiera L, Jones JO, Gopinathan A, Tuveson DA, Fearon DT. Suppression of antitumor immunity by stromal cells expressing fibroblast activation protein-alpha. Science. 2010 Nov 5;330(6005):827-30. Kratochwil C, Flechsig P, Lindner T, Abderrahim L, Altmann A, Mier W, et al. 68Ga-FAPI PET / CT: Tracer uptake in 28 different kinds of cancer. Journal of Nuclear Medicine. 2019;60:801-5. Krishnaraju VS, Kumar R, Mittal BR, Sharma V, Singh H, Nada R et al. Differentiating benign and malignant pancreatic masses: Ga-68 PSMA PET / CT as a new diagnostic avenue. European Radiology. 2021;31:2199-208. Laverman P, van der Geest T, Terry SY, Gerrits D, Walgreen B, Helsen MM, Nayak TK, Freimoser-Grundschober A, Waldhauer I, Hosse RJ, Moessner E, Umana P, Klein C, Oyen WJ, Koenders MI, Boerman OC. Immuno-PET and Immuno-SPECT of Rheumatoid Arthritis with Radiolabeled Anti-Fibroblast Activation Protein Antibody Correlates with Severity of ImmunoPET and immunoSPECT in rheumatoid arthritis using radiolabeled anti-fibroblast activation protein antibody correlates with arthritis severity. J Nucl Med. 2015 May;56(5):778-83. Ling X, Cutler CS, Anderson CJ. The radiopharmaceutical chemistry of the radioisotopes of copper. In:Lewis JS, Windhorst AD, Zeglis BM, editors. Radiopharmaceutical Chemistry. Cham: Springer International Publishing; 2019. p. 335-58. Liu, H.;Moy, P.;Kim, S.;Xia, Y.;Rajasekaran, A.;Navarro, V.;Knudsen, B.;Bander, NH Cancer Res. 1997, 57, 3629. Lo PC, Chen J, Stefflova K, Warren MS, Navab R, Bandarchi B, Mullins S, Tsao M, Cheng JD, Zheng G. Photodynamic molecular beacon triggered by fibroblast activation protein on cancer-associated fibroblasts for diagnosis and treatment of epithelial cancers. J Med Chem. 2009 Jan 22;52(2):358-68. Loktev A, Lindner T, Burger EM, Altmann A, Giesel F, Kratochwil C, et al. Development of fibroblast activation protein-targeted radiotracers with improved tumor retention. Journal of Nuclear Medicine. 2019;60:1421-9. Lopez JI, Errarte P, Erramuzpe A, Guarch R, Cortes JM, Angulo JC, et al. Fibroblast activation protein predicts prognosis in clear cell renal cell carcinoma. Human Pathology. 2016;54:100-5. Luo H, Hernandez R, Hong H, Graves SA, Yang Y, England CG et al., Noninvasive brain cancer imaging with a bispecific antibody fragment, generated via click chemistry. Proceedings of the National Academy of Sciences. 2015;112:12806-11. Meester I, Augustyns K, Van der Veken P. Acylated Gly-(2-cyano)pyrrolidines as inhibitors of fibroblast activation protein (FAP) and the issue of FAP / prolyl oligopeptidase (PREP)-selectivity. Bioorg Med Chem Lett. 2012 May 15;22(10):3412-7. Meyer AR, Carducci MA, Denmeade SR, Markowski MC, Pomper MG, Pierorazio PM, et al. Improved identification of patients with oligometastatic clear cell renal cell carcinoma with PSMA-targeted 18F-DCFPyL PET / CT. Annals of Nuclear Medicine. 2019;33:617-23. Mona CE, Benz MR, Hikmat F, Grogan TR, Luckerath K, Razmaria A, et al. Correlation of 68Ga-FAPi-46 PET biodistribution with FAP expression by immunohistochemistry in patients with solid cancers: a prospective translational exploratory study. Journal of Nuclear Medicine. 2021:jnumed.121.262426. Nimmagadda S, Pullambhatla M, Chen Y, Parsana P, Lisok A, Chatterjee S, et al. Low-level endogenous PSMA expression in nonprostatic tumor xenografts is sufficient for in vivo tumor targeting and imaging. Journal of Nuclear Medicine. 2018;59:486-93. Olszewski RT, Bukhari N, Zhou J, Kozikowski AP, Wroblewski JT, Shamimi-Noori S, et al. NAAG peptidase inhibition reduces locomotor activity and some stereotypes in the PCP model of schizophrenia via group II mGluR. Journal of Neurochemistry. 2004;89:876-85. Paschalis A, Sheehan B, Riisnaes R, Rodrigues DN, Gurel B, Bertan C, et al. Prostate-specific membrane antigen heterogeneity and DNA repair defects in prostate cancer. European Urology. 2019;76:469-78. Pereira BA, Vennin C, Papanicolaou M, Chambers CR, Herrmann D, Morton JP, et al., CAF Subpopulations: A New Reservoir of Stromal Targets in Pancreatic Cancer. Trends in Cancer. 2019;5:724-41. Poels TT, Vuijk FA, de Geus-Oei LF, Vahrmeijer AL, Oprea-Lager DE, Swijnenburg RJ. Molecular Targeted Positron Emission Tomography Imaging and Radionuclide Therapy of Pancreatic Ductal Adenocarcinoma. Cancers. 2021;13:6164. Poplawski SE, Lai JH, Li Y, Jin Z, Liu Y, Wu W, Wu Y, Zhou Y, Sudmeier JL, Sanford DG, Bachovchin WW. Identification of selective and potent inhibitors of fibroblast activation protein and prolyl oligopeptidase. J Med Chem. 2013 May 9;56(9):3467-77. Pure E, Blomberg R. Pro-tumorigenic roles of fibroblast activation protein in cancer: back to the basics. Oncogene. 2018;37:4343-57. Reilly RM, Lam K, Chan C, Levine M. Advancing Novel Molecular Imaging Agents from Preclinical Studies to First-in-Humans Phase I Clinical Trials in Academia-A Roadmap for Overcoming Perceived Barriers. Bioconjugate chemistry. 2015;26:625-632. Rettig WJ, Garin-Chesa P, Healey JH, Su SL, Ozer HL, Schwab M, Albino AP, Old LJ. Regulation and heteromeric structure of the fibroblast activation protein in normal and transformed cells of mesenchymal and neuroectodermal origin.Cancer Res. 1993 Jul 15;53(14):3327-35. Rini BI, Plimack ER, Stus V, Gafanov R, Hawkins R, Nosov D, et al. Pembrolizumab plus Axitinib versus Sunitinib for Advanced Renal-Cell Carcinoma. New England Journal of Medicine. 2019;380:1116-27. Rohrich M, Loktev A, Wefers AK, Altmann A, Paech D, Adeberg S, et al. IDH-wildtype glioblastomas and grade III / IV IDH-mutant gliomas show elevated tracer uptake in fibroblast activation protein-specific PET / CT. European Journal of Nuclear Medicine and Molecular Imaging. 2019;46:2569-80. Roy et al., Design and validation of fibroblast activation protein alpha targeted imaging and therapeutic agents, Theranostics 2020, 10(13), 5778-5789. Ryabtsova O, Jansen K, Van Goethem S, Joossens J, Cheng JD, Lambeir AM, De Meester I, Augustyns K, Van der Veken P. Acylated Gly-(2-cyano)pyrrolidines as inhibitors of fibroblast activation protein (FAP) and the issue of FAP / prolyl oligopeptidase (PREP)-selectivity. Bioorg Med Chem Lett. 2012 May 15;22(10):3412-7. Sanford DG, Bachovchin WW. Identification of selective and potent inhibitors of fibroblast activation protein and prolyl oligopeptidase. J Med Chem. 2013 May 9;56(9):3467-77. Scanlan MJ, Raj BK, Calvo B, Garin-Chesa P, Sanz-Moncasi MP, Healey JH, Old LJ, Rettig WJ. Molecular cloning of fibroblast activation protein alpha, a member of the serine protease family selectively expressed in stromal fibroblasts of epithelial cancers. Proc Natl Acad Sci USA. 1994 Jun 7;91(12):5657-61. Schuelke, N.;Varlamova, OA;Donovan, GP;Ma, D.;Gardner, JP;Morrissey, DM;Arrigale, R. R;Zhan, C;Chodera, AJ;Surowitz, K. G;Maddon, PJ;Heston, WDW;Olson, WC Proc. Natl. Acad. Sci. 2003, 100, 12590. Scott AM, Wiseman G, Welt S, Adjei A, Lee FT, Hopkins W, Divgi CR, Hanson LH, Mitchell P, Gansen DN, Larson SM, Ingle JN, Hoffman EW, Tanswell P, Ritter G, Cohen LS, Bette P, Arvay L, Amelsberg A, Vlock D, Rettig WJ, Old LJ. A Phase I dose-escalation study of sibrotuzumab in patients with advanced or metastatic fibroblast activation protein-positive Phase I dose-escalation study of sibrotuzumab in patients with advanced or metastatic fibroblast activation protein-positive cancer. Clin Cancer Res. 2003 May;9(5):1639-47. Shallal HM, Minn I, Banerjee SR, Lisok A, Mease RC, Pomper MG. Heterobivalent agents targeting PSMA and integrin-αvβ3. Bioconjugate Chemistry. 2014;25:393-405. Sheridan C. Amgen's bispecific antibody puffs across finish line. Nature Biotechnology. 2015;33:219-21. Silver DA, Pellicer I, Fair WR, Heston WD, Cordon-Cardo C. Prostate-specific membrane antigen expression in normal and malignant human tissues. Clinical Cancer Research. 1997;3:81-5. Siva S, Udovicich C, Tran B, Zargar H, Murphy DG, Hofman MS. Expanding the role of small-molecule PSMA ligands beyond PET staging of prostate cancer. Nature Reviews Urology. 2020;17:107-18. Slania SL, Das D, Lisok A, Du Y, Jiang Z, Mease RC et al., Imaging of Fibroblast Activation Protein in Cancer Xenografts Using Novel (4-Quinolinoyl)-glycyl-2-cyanopyrrolidine- Based Small Molecules. Journal of Medicinal Chemistry. 2021;64:4059-70. Solano-Iturri JD, Beitia M, Errarte P, Calvete-Candenas J, Etxezarraga MC, Loizate A, et al. Altered expression of fibroblast activation protein-alpha; (FAP) in colorectal adenomacarcinoma sequence and in lymph node and liver metastases. Aging. 2020a;12:10337-58. Solano-Iturri JD, Errarte P, Etxezarraga MC, Echevarria E, Angulo J, Lopez JI, et al. Altered tissue and plasma levels of fibroblast activation protein-α (FAP) in renal tumours. Cancers. 2020b;12:3393. Spatz S, Tolkach Y, Jung K, Stephan C, Busch J, Ralla B, et al. Comprehensive evaluation of prostate specific membrane antigen expression in the vasculature of renal tumors: Implications for imaging studies and prognostic role. Journal of Urology. 2018;199:370-7. Stock K, Steinestel K, Wiesch R, Mikesch JH, Hansmeier A, Trautmann M, et al. Neovascular Prostate-Specific Membrane Antigen Expression Is Associated with Improved Overall Survival under Palliative Chemotherapy in Patients with Pancreatic Ductal Adenocarcinoma. BioMed Research International. 2017;2017:2847303. Tsai TY, Yeh TK, Chen X, Hsu T, Jao YC, Huang CH, Song JS, Huang YC, Chien CH, Chiu JH, Yen SC, Tang HK, Chao YS, Jiaang WT. Substituted 4-carboxymethylpyroglutamic acid diamides as potent and selective inhibitors of fibroblast activation protein. J Med Chem. 2010 Sep 23;53(18):6572-83. Tuxhorn JA, Ayala GE, Smith MJ, Smith VC, Dang TD, Rowley DR. Reactive stroma in human prostate cancer: induction of myofibroblast phenotype and extracellular matrix remodeling. Clin Cancer Res. 2002 Sep;8(9):2912-23. U.S. Patent Application Publication No. US2014 / 0357650, Novel FAP Inhibitors, to Jansen et al., published December 4, 2014. U.S. Patent No. 9,346,814, for Novel FAP Inhibitors, to Jansen et al., issued May 24, 2016. Uijen MJM, Derks YHW, Merkx RIJ, Schilham MGM, Roosen J, Prive BM, et al., PSMA radioligand therapy for solid tumors other than prostate cancer: background, opportunities, challenges, and first clinical reports. European Journal of Nuclear Medicine and Molecular Imaging. 2021;48:4350-68. Watabe T, Liu Y, Kaneda-Nakashima K, Shirakami Y, Lindner T, Ooe K et al. Theranostics targeting fibroblast activation protein in the tumor stroma: 64Cu- and 225Ac-labeled FAPI-04 in pancreatic cancer xenograft mouse models. Journal of Nuclear Medicine. 2020;61:563-9. Welt S, Divgi CR, Scott AM, Garin-Chesa P, Finn RD, Graham M, Carswell EA, Cohen A, Larson SM, Old LJ, et al. Antibody targeting in metastatic colon cancer: a phase I study of monoclonal antibody F19 against a cell-surface protein of reactive tumor stromal fibroblasts. J Clin Oncol. 1994 Jun;12(6):1193-203. Wernicke AG, Kim S, Liu H, Bander NH, Pirog EC. Prostate-specific membrane antigen (PSMA) expression in the neovasculature of gynecologic malignancies: implications for PSMA-targeted therapy. Applied Immunohistochemistry & Molecular Morphology. 2017;25:271-6. Youn H, Hong K. In vivo noninvasive small animal molecular imaging. Osong Public Health Res Perspect. 2012;3 :48-59. PMCID:PMC3738683 Yu DM, Yao TW, Chowdhury S, Nadvi NA, Osborne B, Church WB, McCaughan GW, Gorrell MD. The dipeptidyl peptidase IV family in cancer and cell biology. FEBS J. 2010 Mar;277(5):1126-44. Zhang et al., Prostate-specific membrane antigen-targeted photoacoustic imaging of prostate cancer in vivo, J. of Biophotonics, 2018;11:e201800021. Zhou L, Chang Y, Xu L, Liu Z, Fu Q, Yang Y, et al. The Presence of Vascular Mimicry Predicts High Risk of Clear Cell Renal Cell Carcinoma after Radical Nephrectomy. Journal of Urology. 2016;196:335-42.

[0456] Although the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications may be practiced within the scope of the appended claims.

Claims

1. Formula (III): 【Chemistry 1】 and stereoisomers and pharma- ceutically acceptable salts thereof; During the ceremony: A is a targeting moiety for fibroblast activation protein alpha (FAP-α); B is absent or is a targeting moiety for FAP-α, and A and B can be the same or different; C 1 may be absent or present, and when present is a chelating group; C 2 is a prosthetic group; the prosthetic group C2 comprises a radioisotope selected from the group consisting of 18F, 124I, 125I, 131I, and 211At, and / or C1 is present and comprises a radiometal; L a and L b are bifunctionalized linkers each capable of forming chemical bonds with each other and with A, B, Lc1, and Lc2; L c1 and L c2 are independent of each other and C 1, C 2, L a , and L b a bifunctional linker capable of forming a chemical bond with; C 1 If absent, L c1 is also absent, and if B is absent, then L b is also absent, as well as stereoisomers and pharma- ceutically acceptable salts thereof.

2. A has the structure: 【Chemistry 2】 or when B is present, A and B each have the formula: During the ceremony, 【Chemistry 3】 is the linker L a and / or L b indicates the point of attachment of the FAP-α binding ligand to, which may be at any carbon atom 5, 6, 7, or up to 8 of the quinolinyl ring; each y is independently an integer selected from the group consisting of 0, 1, and 2; R 1x , R 2x , and R 3x′ are each independently selected from the group consisting of H, OH, halogen, C 1-6 alkyl, —OC 1-6 alkyl, and —SC 1-6 alkyl; 2. The compound of claim 1 and its stereoisomers and pharma- ceutically acceptable salts.

3. C 2 teeth 18 F, 124 I, 125 I, 131 I, and 211 2. The compound of claim 1, comprising a radioisotope selected from the group consisting of At.

4. The prosthetic groups are: 【Chemistry 4】 Selected from the group consisting of: where each X is a straight chain or branched C 1 -C 8 Alkyl, -SO 2 , -C(=O)-, -C(=O)OR 20 are independently selected from the formula R 20 is H or C 1 -C 4 Alkyl, and 18 F, 124 I, 125 I, 131 I, and 211 each R and R' is defined above; and each n is independently an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, wherein 【Chemistry 5】 Each carbon in the alkylene chain is C 1 -C 4 The compound of claim 1, which can be substituted with alkyl.

5. The prosthetic groups are: 【Chemistry 6】 5. The compound of claim 4 selected from the group consisting of:

6. (a)C 1 These include DOTAGA (1,4,7,10-tetraazacyclododecane-1-(glutaric acid)-4,7,10-triacetic acid), DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTASA (1,4,7,10-tetraazacyclododecane-1-(2-succinic acid)-4,7,10-triacetic acid), CB-DO2A (10-bis(carboxymethyl)-1,4,7,10-tetraazabicyclo[5.5.2]tetradecane), DEPA (7-[2-(bis-carboxymethylamino)-ethyl]-4,10-bis-carboxymethyl-1,4,7,10-tetraaza- cyclododec-1-yl-acetic acid), 3p-C-DEPA (2-[(carboxymethyl)][5-(4-nitrophenyl-1-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl]pentan-2-yl)amino]acetic acid), TCMC (2-(4-isothiocyanotobenzyl)-1,4,7,10-tetraaza-1,4,7,10-tetra-(2-carbamonylmethyl)-cyclododecane), oxo-DO3A (1-oxa-4,7,10-triazacyclododecane-5-S-(4-isothiocyanatobenzyl)-4,7,10-triacetic acid), p-NH 2 -Bn-oxo-DO3A (1-oxa-4,7,10-tetraazacyclododecane-5-S-(4-aminobenzyl)-4,7,10-triacetic acid), TE2A (1,8-N,N′-bis-(carboxymethyl)-1,4,8,11-tetraazacyclotetradecane), MM-TE2A, DM-TE2A, CB-TE2A (4,11-bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane), CB-TE1A1P (4,8,11-tetraazacyclotetradecane) -1-(methanephosphonic acid)-8-(methanecarboxylic acid), CB-TE2P (1,4,8,11-tetraazacyclotetradecane-1,8-bis(methanephosphonic acid)), TETA (1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid), NOTA (1,4,7-triazacyclononane-N,N′,N″-triacetic acid), NODA (1,4,7-triazacyclononane-1,4-diacetate); NODAGA (1,4,7-triazacyclononane-1-glutaric acid-4,7- acetic acid), NOTAGA ((1,4,7-triazonane-1,4-diyl)diacetic acid), DFO (deferoxamine), NETA ({4-[2-(bis-carboxymethylamino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid), TACN-TM (N,N',N''-tris(2-mercaptoethyl)-1,4,7-triazacyclononane), Diamsar (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo(6,6,6)eicosane- 3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), Sarar (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine), AmBaSar (4-((8-amino-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosan-1-ylamino)methyl)benzoic acid), and BaBaSar; or (b) C1 is: 【Chemistry 7-1】 【Chemistry 7-2】 【Chemistry 7-3】 【Chemistry 7-4】 2. The compound of claim 1, which is a chelating agent selected from the group consisting of:

7. C 1 is 【Chemistry 8】 2. The compound of claim 1 comprising:

8. The compound of claim 1, wherein C1 is present and, optionally, the radiometal is selected from the group consisting of 60Cu, 62Cu, 64Cu, 67Cu, 203Pb, 212Pb, 225Ac, 177Lu, 99mTc, 68Ga, 149Tb, 86Y, 90Y, 111In, 115In, 186Re, 188Re, 153Sm, 89Zr, 213Bi, 212Bi, 212Pb, 67Ga, 47Sc, 166Ho, 43Sc, 223Ra, 226 / 227Th, Al-18F, and Sc-18F.

9. L a , L b , L c1 , and L c2 are each independently selected from the group consisting of (a), (b), (c), or (d): （a） 【Chemistry 9】 and During the ceremony: p 1 , p 2 , p 3 and p 4 may be in any order; t 1 and t 2 are each an integer selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; p 1、 p 3 , and p 4 are each independently 0 or 1; p 2 is an integer selected from the group consisting of 0, 1, 2, and 3, and p 2 When is 2 or 3, each R 1 are the same or different; m 1 , m 2 , m 3 , and m 4 are each an integer independently selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, and 8; W 1 is selected from the group consisting of a bond, -S-, -C(=O)-, -C(=O)-NR-, and -NR-C(=O)-; W 2 is a bond, -S-, -CH 2 -C(=O)-NR-, -C(=O)-, -NRC(=O)-, -NR'C(=O)NR-, -NRC(=S)NR' 2 -, -NRC(=O)O-, -OC(=O)NR-, -OC(=O)-, -C(=O)NR-, -NR-C(=O)-, -C(=O)O-, -(O-CH 2 -CH 2 ) q - and -(CH 2 -CH 2 -O) q wherein q is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; Each R or R' is independently H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, and -OR 4 where R 4 is selected from the group consisting of H, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, and substituted heterocycloalkyl; H can be present or absent, and when present, is a nitrogen-containing heteroalkylene chain, a cyclic or bicyclic heteroalkyl radical, and the structure 【Chemistry 10】 A triazole radical having the formula: Each R 1 Regardless of H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, -(CH 2 ) q -C 3 -C 12 Aryl, -C 4 -C 16 Alkylaryl, or -(CH 2 ) q -C 4 -C 16 alkylaryl; R 2 and R 3 are independently H, -(CH 2 ) q -C 3 -C 12 Aryl, and -CO 2 R 5 where R 5 is H, C 1 -C 6 Alkyl, C 3 -C 12 Aryl, and C 4 -C 16 alkylaryl; R 2 and R 3 can be the same or different, and q is selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; V is selected from the group consisting of -C(O)-, -C(S)-, -NRC(O)-, -NRC(S)-, and -OC(O)-; （b） 【Chemistry 11】 and In the formula, t 2 , p 1 , p 2 , p 3 , m 1 , m 2 , W 2 , R, R 1 , R 2 , R 3 and V is defined above; Tz is a triazole group which may be present or absent, and when present, 【Chemistry 12】 Selected from the group consisting of: (c)-L 1 -, -L 2 -L 3 - or -L 1 -L 2 -L 3 -, wherein: L 1 -NR-(CH 2 ) q -[O-CH 2 -CH 2 -O] q -(CH 2 ) q -C(=O)-; L 2 -NR-(CH 2 ) q -C(COOR 5 )-NR-; and L 3 is -(O=)C-(CH 2 ) q -C(=O)-; wherein each q is independently an integer selected from the group consisting of 1, 2, 3, 4, 5, 6, 7, and 8; and R and R 5 is as defined above; and (d)-(CR 6 H) q -(CH 2 ) q -C(=O)-NR-(CH 2 ) q -O- or -NR-(CH 2 ) q -O-; wherein each q, R5 and R are defined above; and R 6 H or -COOR 5 and Optionally, (a) H is: 【Chemistry 13】 wherein X 1 and X 2 are each independently -CH- or N; each R 16 is independently H or -C(═O)-OR 17 , where R 17 is C 1 -C 4 alkyl; -N(R18)-(CH2)z5-N(R19)-; where R18 and R19 are each independently H or C1-C4 alkyl, and Z5 is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; 【Chemistry 14】 or (b) one or more of L a , L b , L c1 , and L c2 are: 【Chemistry 15-1】 【Chemistry 15-2】 It includes one or more units selected from; 2. The compound of claim 1, wherein u is an integer selected from 1, 2, 3, 4, 5, 6, 7, and 8; and R and R5 are as defined above.

10. L a is 【Chemistry 16】 2. The compound of claim 1,

11. L c1 is 【Chemistry 17】 and u is 1, Lc2 is 【Chemistry 18】 2. The compound of claim 1,

12. The compound of formula (III) is as follows: 【Chemistry 19-1】 【Chemistry 19-2】 【Chemistry 19-3】 The compound of claim 1 selected from:

13. The compound of formula (III) 【Chemistry 20】 and optionally, F includes the radioisotope 18 F, or the radiometal of the chelating group is selected from the group consisting of 60Cu, 62Cu, 64Cu, 67Cu, 203Pb, 212Pb, 225Ac, 177Lu, 99mTc, 68Ga, 149Tb, 86Y, 90Y, 111In, 115In, 186Re, 188Re, 153Sm, 89Zr, 213Bi, 212Bi, 212Pb, 67Ga, 47Sc, 166Ho, 43Sc, 223Ra, 226 / 227Th, Al-18F, and Sc-18F; or F comprises the radioisotope 18F and the radiometal of the chelating group is selected from the group consisting of 60Cu, 62Cu, 64Cu, 67Cu, 203Pb, 212Pb, 225Ac, 177Lu, 99mTc, 68Ga, 149Tb, 86Y, 90Y, 111In, 115In, 186Re, 188Re, 153Sm, 89Zr, 213Bi, 212Bi, 212Pb, 67Ga, 47Sc, 166Ho, 43Sc, 223Ra, 226 / 227Th, Al- 18F, and Sc- 18F; The compound of claim 1.

14. The compound of formula (III) 【Chemistry 21】 2. The compound of claim 1, wherein, optionally, F comprises the radioactive isotope 18F.

15. prosthetic group C 2 is a chelating group C 1 covalently binds to Optionally, the following: 【Chemical 22】 The compound of claim 1 selected from:

16. A compound according to any one of claims 1 to 15, Optionally, the pharmaceutical composition further comprises one or more pharma- ceutically acceptable carriers, diluents, excipients, or adjuvants.

17. A compound described in any one of claims 1 to 15, or a pharmaceutical composition described in claim 16, for use in a method for imaging a disease or disorder associated with fibroblast activation protein-alpha (FAP-α) and / or prostate-specific membrane antigen (PSMA), wherein the compound includes any optically or radiolabeled functional group suitable for optical imaging, photoacoustic imaging, PET imaging, or SPECT imaging, and the method comprises administering the compound or pharmaceutical composition and obtaining an image.

18. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use as a medicine.

19. A compound according to any one of claims 1 to 15, or a pharmaceutical composition according to claim 16, for use in treating a fibroblast activation protein-alpha (FAP-α)- and / or prostate-specific membrane antigen (PSMA)-related disease or disorder, wherein the compound comprises a radiolabel functional group suitable for radiotherapy, and the compound or pharmaceutical composition is administered in an effective amount to a subject in need of treatment, and optionally (i) the (FAP-α)-related disease or disorder is selected from the group consisting of proliferative diseases; diseases characterized by tissue remodeling and / or chronic inflammation; disorders associated with endocrine dysfunction; and blood clotting disorders, and optionally, the proliferative disease is selected from the group consisting of breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, melanoma, fibrosarcoma, bone and connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, and adenocarcinoma, or (ii) the compound or pharmaceutical composition wherein the prostate-specific membrane antigen (PSMA)-associated disease or disorder is selected from the group consisting of prostate cancer, kidney cancer, head cancer, neck cancer, lung cancer, breast cancer, colorectal cancer, esophageal cancer, gastric cancer, leukemia / lymphoma, uterine cancer, skin cancer, endocrine cancer, urinary cancer, pancreatic cancer, digestive cancer, ovarian cancer, cervical cancer, adenoma, and tumor neovasculature, and optionally, the prostate-specific membrane antigen (PSMA)-associated disease or disorder includes prostate cancer.

Images