Methods and materials for combining biologics with multiple chelators - Patents.com

Abstract

Provided herein are conjugates that include two or more chelators (e.g., a radiotherapeutic isotope chelator and an imaging isotope chelator) covalently attached to one or more binding moieties. The conjugates can be used to treat cancerous or non-cancerous conditions, and can act as both imaging and radiotherapeutic molecules when an imaging isotope is complexed to an imaging isotope chelator and a radiotherapeutic isotope is complexed to a radiotherapeutic isotope chelator.

Description

[Technical field]

[0001] (CROSS REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Patent Application No. 63 / 211,919, filed June 17, 2021. The disclosure of this prior application is considered part of (and incorporated by reference into) the disclosure of this application.

[0002] background 1. Technical Field This document relates to conjugates of two or more chelators (e.g., conjugates of a chelator of an imaging isotope and a chelator of a radiotherapeutic isotope) with one or more binding moieties, and the use of such conjugates to treat diseases such as cancer. For example, this document provides methods and materials for combining a binding moiety with two or more chelators, where one of the chelators is a chelator of an isotope used for imaging and one of the chelators is a chelator of an isotope used for radiotherapy. The conjugates in which the imaging isotope and the radiotherapeutic isotope are complexed to the chelator can be administered to a mammal in need of treatment and can act as both an imaging molecule and a radiotherapeutic molecule. [Background technology]

[0003] 2. Background information In the field of targeted radionuclide therapy, the ability to accurately calculate dosimetry (how much therapeutic agent reaches tumors and tissues in the body) through patient imaging is an effective way to understand disease pathology, disease progression, and response to radionuclide therapy, and also helps enhance drug discovery and personalize care for patients (e.g., cancer patients) through better understanding of pharmacokinetics and pharmacodynamics, facilitating regulatory (e.g., FDA) approval. The field of targeted radionuclide therapy is moving toward more effective and often more expensive alpha emitters and away from beta emitters. However, alpha emitters are typically not suitable for imaging due to the unavailability or low doses of suitable positron or photon energy emissions (511 KeV for PET, 100-200 KeV for SPECT). Alpha emissions with high linear energy transfer (LET) and bremsstrahlung radiation with gamma photons, characteristic x-rays, or decay of the parent alpha-emitting nuclide are poorly suited for quantification of target uptake, dosimetry, and treatment response compared to beta-emitters. Furthermore, even when treatment is performed with imageable beta-emitters, the beta-emitters are often poorly imaged by SPECT techniques. If radionuclide therapy could be imaged by PET techniques, the image resolution, accuracy, and quality would be much better. As a result, most research and development, FDA submissions, and clinical programs must rely on estimated biodistribution / dosimetry based on poor quality images or by using surrogate imaging probes (imageable modified drugs). These surrogate imaging probes are significantly different from alpha-emitter therapeutics in a variety of ways, making them poor predictors of the biodistribution / dosimetry of alpha-emitting therapeutics. Thus, improved radiation therapy that can be directly and accurately imaged is needed. Summary of the Invention [Means for solving the problem]

[0004] overview This document is based at least in part on the discovery of a method to combine (e.g., covalently link) multiple chelators with a binding moiety or motif, e.g., a biologic or drug that binds to a target molecule in a mammal, such that the resulting conjugate or mixture of conjugates can simultaneously serve as both an imaging molecule and a radiotherapy molecule when a suitable isotope is complexed with the chelator. The resulting conjugate comprises two or more chelators and a binding moiety (e.g., two or more chelators covalently linked to the binding moiety via one or more linkers), where one of the chelators is a chelator of an isotope used for imaging (referred to herein as a "chelator of an imaging isotope") and one of the chelators is a chelator of an isotope used for radiotherapy (referred to herein as a "chelator of a radiotherapy isotope"). As described herein, the conjugates can be selectively used for imaging or radionuclide therapy, as desired, by selecting an imaging radionuclide or a therapeutic radionuclide and loading the other chelator with a non-radioactive imaging metal ion or a therapeutic metal ion to maintain the same chemical nature of the molecule. By using the same chemical entity, one maintains the same biodistribution and avoids the use of alternative imaging probes that are structurally different and may have different biodistributions. Furthermore, by complexing both chelators with appropriate imaging and therapeutic radionuclides, the same conjugate can be used for both imaging and radionuclide therapy, without being forced to select only a single isotope that is suboptimal for one or both tasks.

[0005] The conjugates and methods described herein can allow for biodistribution and dosimetry of alpha-emitting therapeutics to be assessed pre-treatment and with each cycle of radiation therapy, helping to accelerate research and development, expedite FDA approval, and guide clinical care. Additionally, the methods described herein can be used to streamline the ongoing evaluation of patients undergoing these costly radiation treatments with more accurate treatment monitoring (e.g., by imaging the treatment immediately after it is administered) and can also be streamlined with simplified clinical workflow. This can result in informed changes in the treatment care plan, saving money by stopping futile treatments earlier, improving outcomes by adjusting or boosting treatments as needed, or switching to more effective treatments more quickly.

[0006] The conjugates described herein can be designed to match the half-life of the imaging isotope (e.g., an isotope for positron emission tomography (PET) or an isotope for single photon emission computed tomography (SPECT)) with the physical half-life of the radiotherapeutic isotope (e.g., an alpha- or beta-emitting radionuclide) to ensure that the biodistribution of the therapy over the time that it is radioactive can be imaged and thus dosimetry can be accurately calculated. For example, the half-life of the imaging isotope (e.g., an isotope for PET or an isotope for SPECT), the physical half-life of the radiotherapeutic isotope (e.g., an alpha- or beta-emitting radionuclide), and the plasma half-life of the targeting vector (e.g., a peptide, antibody, or small molecule) can be matched to ensure that the biodistribution of the therapy over the time that it is radioactive can be imaged and thus dosimetry can be accurately calculated. In some embodiments, an optical imaging (near infrared) probe can be added to the conjugate. The conjugates and methods described herein provide a robust platform to stage disease, treat disease, monitor response or progression to treatment, and / or minimize side effects on healthy organs and tissues, all using the same (chemically and biologically identical) molecular type. This can be achieved simply by choosing whether the conjugates described herein are complexed with imaging isotopes and / or with radiotherapeutic isotopes or non-radioactive versions of these same isotopes (i.e., radionuclides can be exchanged with chemically identical non-radioactive isotopes having different nuclear structures) for the desired use of the conjugate. In some embodiments, more than one conjugate can be used. For example, in some embodiments, one conjugate described herein is complexed with an alpha-emitting isotope for therapy and one conjugate described herein is complexed with a positron-emitting isotope for imaging. Additionally, the conjugates described herein can include more than one binding moiety or motif to enhance uptake in targeted tissues / organs.

[0007] In one general aspect, the present disclosure provides a conjugate comprising two or more chelators and a binding moiety, wherein one of the chelators is a chelator for an imaging isotope and one of the chelators is a chelator for a radiotherapeutic isotope.

[0008] In some embodiments, the isotope used for the radiation therapy is an alpha emitter, hi some embodiments, the isotope used for the radiation therapy is both an alpha emitter and a beta emitter.

[0009] In some embodiments, the radiotherapeutic isotope is 225 Ac, 212 Pb, 211 At, 213 Bi, 212 Bi, 211 Bi, 227 Th, 223 Ra, 211 Po, 221 Fr, 217 At, 213 Po, 212 Po, 215 Po, or 177 In some embodiments, the radiotherapeutic isotope is 225 Ac, 212 Pb, 211 At, 213 Bi, 212 Bi, 211 Bi, 152 / 160 / 161 Tb, 227 Th, 223 Ra, 211 Po, 221 Fr, 217 At, 213 Po, 212 Po, 215 Po, or 177 This is Lu.

[0010] In some embodiments, the imaging isotope is 68 Ga, 44 Sc, 60 / 61 / 62 / 64 Cu, 84 / 86 / 87 / 89 Zr, 63 Zn,43 / 44 Sc, 192 / 193 / 194 / 196 Au, 52m Mn, 90 / 92m1 Nb, 51 / 52 Mn, 45 Ti, 65 / 66 Ga, 94m Tc, 55 Co, 80 / 81 / 83 Sr, 38 K, 70 / 71 / 72 / 74 As, 81 / 82m Rb, 52 Fe, or 86 In some embodiments, the imaging isotope is 68 Ga, 44 Sc, 60 / 61 / 62 / 64 Cu, 84 / 86 / 87 / 89 Zr, 63 Zn, 43 / 44 Sc, 192 / 193 / 194 / 196 Au, 52m Mn, 90 / 92m1 Nb, 51 / 52 Mn, 148 / 151 / 151m / 152 Tb, 45 Ti, 65 / 66 / 67 Ga, 94m Tc, 55 Co, 80 / 81 / 83 Sr, 38 K, 70 / 71 / 72 / 74 As, 81 / 82m Rb, 52 Fe, or 86 It's Y.

[0011] In some embodiments, the imaging isotope is 64 Cu, and said radiotherapeutic isotope is 212 It is Pb.

[0012] In some embodiments, the imaging isotope is complexed to the chelator of the imaging isotope.

[0013] In some embodiments, the radiotherapeutic isotope is complexed to the chelator of the radiotherapeutic isotope.

[0014] In some embodiments, each of the chelators is independently selected from 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), dodecanetetraacetic acid (DOTA), 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC), 1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), and diethylenetraminepentacetic acid. acid) (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA). In some embodiments, each of the chelators independently comprises a compound selected from the group consisting of NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, 2,2',2"-nitrilotriacetic acid; (NTA), 2,2-bis(hydroxymethyl)-2,2',2"-nitrilotriethanol (BisTris), ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA), 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), 1,4,7,10-tetraazacyclododecane-1,7-diacetic acid (DO2A), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid (DO3A), and MACROPA.

[0015] In some embodiments, the binding moiety is a polypeptide.

[0016] In some embodiments, the polypeptide binds to prostate specific membrane antigen, somatostatin receptor, fibroblast activation protein, or melanocortin-1 receptor.

[0017] In some embodiments, the polypeptide is an antibody.

[0018] In some embodiments, the binding moiety is a small molecule.

[0019] In some embodiments, the small molecule is a glutamate carboxypeptidase II inhibitor.

[0020] In some embodiments, the chelator is covalently attached to the binding moiety.

[0021] In some embodiments, the chelator and the binding moiety are covalently attached via a linker.

[0022] In some embodiments, the chelator and the binding moiety are represented by the moiety of formula (I):

[0023] [ka] (In the formula: Each X is N, P, P(=O), CR N and a moiety of formula (i): [ka] are independently selected from; each of x1, x2, x3, and x4 independently represents a point of attachment of the moiety of formula (I) to a chelator or binding moiety; L 1 , L 2 , L 3 , and L 4 are C(=O), C(=S), and N(R N ), O, S, S(=O), S(=O)2, -CR N =NR N -, (-C 1-3 Alkylene-O-) x , (-OC 1-3 Alkylene-) x , -C 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, where each x is independently an integer from 1 to 10; 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 Each of arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from OH, NO, CN, halo, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Haloalkoxy, Amino, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 optionally substituted with 1, 2, or 3 substituents independently selected from alkoxycarbonyl; Each of y1, y2, y3, and y4 is independently an integer from 1 to 10; Each R N , H, C 1-3 Alkyl, and C 1-3 independently selected from haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5. are connected via.

[0024] In some embodiments, the moiety of formula (I) has any one of the following formulas:

[0025] [ka] I have TIFF2024523344000005.tif145160.

[0026] In some embodiments, the chelator and the binding moiety are a moiety of formula (II):

[0027] [ka] (In the formula, x1 represents the point of attachment of formula (II) to the chelator; x2 represents the point of attachment of formula (II) to a chelator or binding moiety; Each L is C(=O), C(=S), N(R N ), O, S, S(=O), S(=O)2, -CR N =NR N -, (-C 1-3 Alkylene-O-) x , (-OC 1-3 Alkylene-) x , -C 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, where each x is independently an integer from 1 to 10; 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 Each of arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from OH, NO, CN, halo, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Haloalkoxy, Amino, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 optionally substituted with 1, 2, or 3 substituents independently selected from alkoxycarbonyl; y is an integer from 1 to 30; Each R N , H, C 1-3 Alkyl, and C 1-3 haloalkyl) are connected via

[0028] In some embodiments, the moiety of formula (II) has any one of the following formulae:

[0029] [ka] has.

[0030] In another general aspect, this document provides a method of treating cancer in a mammal in need of such treatment, the method comprising administering to the mammal a conjugate described herein, the conjugate comprising the imaging isotope complexed to the chelator of the imaging isotope, and the conjugate comprising the radiotherapeutic isotope complexed to the chelator of the radiotherapeutic isotope.

[0031] In another general aspect, this document provides a method of treating cancer in a mammal, the method comprising: a) administering to said mammal a first conjugate comprising two or more chelators and a binding moiety, wherein one of said chelators is a chelator for an imaging isotope and one of said chelators is a chelator for a radiotherapeutic isotope, said first conjugate comprising said imaging isotope complexed to said chelator for said imaging isotope; b) determining the biodistribution of the first conjugate in the mammal; and c) administering to said mammal an amount of said second conjugate, said second conjugate being identical to said first conjugate except that said second conjugate comprises said radiotherapeutic isotope complexed to said chelator of said radiotherapeutic isotope. Includes.

[0032] In some embodiments, the method further comprises determining in the mammal the biodistribution of the second conjugate comprising the imaging isotope complexed to the chelator of the imaging isotope and the radiotherapeutic isotope complexed to the chelator of the radiotherapeutic isotope.

[0033] In some embodiments, the cancer is selected from the group consisting of prostate cancer, neuroendocrine cancer, colon cancer, lung cancer, pancreatic cancer, melanoma, and lymphatic cancer.

[0034] In another general aspect, this document provides a method of treating cancer in a mammal in need of such treatment, the method comprising administering to the mammal two or more conjugates: each conjugate comprises two or more chelators and a binding moiety, one of the chelators being a chelator for an imaging isotope and one of the chelators being a chelator for a radiotherapeutic isotope; one of the conjugates administered to the mammal comprises an imaging isotope complexed to the chelator of the imaging isotope; One of the conjugates administered to the mammal comprises a radiotherapeutic isotope complexed to the chelator of the radiotherapy isotype.

[0035] In some embodiments, the conjugate comprises two or more binding moieties. In some embodiments, the binding moieties can be polypeptides. In some embodiments, each of the polypeptides can independently bind to prostate specific membrane antigen, somatostatin receptor, fibroblast activation protein, or melanocortin-1 receptor.

[0036] In some embodiments, the conjugate comprises three or more chelators, each of which may independently comprise a compound selected from the group consisting of NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, DFO, NTA, BisTris, EGTA, EDTA, BAPTA, DO2A, DTPA, DO3A, and MACROPA.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention pertains. To carry out the present invention, methods and materials similar or equivalent to those described herein can be used, but suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0038] The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. [Brief description of the drawings]

[0039] [Figure 1] Figure 1 shows the attenuation scheme for 212Pb. [Figure 2A] FIG. 2A is an example of a conjugate of two or more chelators attached to a binding moiety. [Figure 2B] FIG. 2B is an example of a conjugate of two or more chelators attached to a binding moiety. [Diagram 3] FIG. 3 is a scheme of the diamsar (Cu) and TCMC (Pb) platforms for peptide conjugation. [Figure 4]FIG. 4 is a scheme of the NOTA (Cu) and TCMC (Pb) platforms for peptide conjugation. [Diagram 5] FIG. 5 is a scheme of the diamsar (Cu) and TCMC (Pb) platforms for dual peptide conjugation. [Figure 6] FIG. 6 is a scheme of the NOTA (Cu) and TCMC (Pb) platform for dual peptide conjugation. [Figure 7] FIG. 7 is a representative example of the synthesis of NOTA (Cu), TCMC (Pb) and peptide (PSMA) conjugates with different linker molecules. [Figure 8] FIG. 8 is a representative example of the synthesis of diamsar (Cu), TCMC (Pb) and peptide (PSMA) conjugates with different linker molecules. [Figure 9] FIG. 9 is an example of a conjugate with a linear arrangement of chelator and binding moiety using the diamsar (Cu) and TCMC (Pb) platforms for peptide conjugation. [Figure 10] FIG. 10 is a high performance liquid chromatography (HPLC) trace of a conjugate (eg, conjugate 1) containing NOTA (Cu) and TCMC (Pb) with an aniline linker. [Figure 11] FIG. 11 is a graph of the HPLC calibration curve for Conjugate 1. [Figure 12] FIG. 12 is an HPLC trace of unlabeled 64Cu. [Figure 13] FIG. 13 is a thin layer chromatography (TLC) trace of unlabeled 64Cu. [Figure 14] FIG. 14 is an example of labeling conjugate 1 with 64Cu to form 64Cu-conjugate-1. [Figure 15] FIG. 15 is a TLC trace of 64Cu-conjugate 1. [Figure 16] FIG. 16 is an HPLC trace of 64Cu-conjugate 1. [Figure 17] FIG. 17 is an HPLC trace of a conjugate (eg, conjugate 2) containing NOTA (Cu) and TCMC (Pb) with an amino acid linker. [Figure 18] FIG. 18 is an example of labeling conjugate 2 with 64Cu to form 64Cu-conjugate 2. [Figure 19] FIG. 19 is a graph of the HPLC calibration curve for Conjugate 2. [Figure 20] FIG. 20 is a TLC trace of 64Cu-conjugate 2. [Figure 21] FIG. 21 is an HPLC trace of 64Cu-conjugate 2. [Figure 22] FIG. 22 is an HPLC trace of 64Cu-conjugate 2 after 40 minutes. [Diagram 23] FIG. 23 is an HPLC trace of 64Cu-conjugate 2 after 2 hours. [Figure 24] FIG. 24 is an HPLC trace of 64Cu-conjugate 2 after 4 hours. [Diagram 25] FIG. 25 is an HPLC trace of 64Cu-conjugate 2 after 8 hours. [Figure 26] FIG. 26 is a TLC trace of 64Cu-conjugate 2 after 40 minutes. [Figure 27] FIG. 27 is a TLC trace of 64Cu-conjugate 2 after 2 hours. [Figure 28] FIG. 28 is a TLC trace of 64Cu-conjugate 2 after 4 hours. [Figure 29] FIG. 29 is a TLC trace of 64Cu-conjugate 2 after 8 hours. [Diagram 30] FIG. 30 is a graph of the percentage of cellular uptake of 64Cu-conjugate 2 with and without inhibitor. [Diagram 31] FIG. 31 is a graph of the standard uptake value (SUV) of 64Cu-conjugate 2 in organs of nude mice. [Diagram 32]FIG. 32 is a graphical blow-up of the SUV of 64Cu-conjugate 2 in nude mouse organs. [Diagram 33] FIG. 33 includes microPET images of normal mice injected with 64Cu-conjugate 2 at different time intervals. [Diagram 34] FIG. 34 is an in vivo PET image of proximal tubules in the kidney of a nude mouse injected with 64Cu-conjugate 2. [Diagram 35] FIG. 35 is an HPLC trace of unlabeled 203Pb. [Diagram 36] FIG. 36 is a TLC trace of unlabeled 203Pb. [Figure 37] FIG. 37 is an example of labeling conjugate 1 with 203Pb to form 203Pb-conjugate-1. [Figure 38] FIG. 38 is an HPLC trace of 203Pb-conjugate 1. [Figure 39] FIG. 39 is an example of labeling conjugate 2 with 203Pb to form 203Pb-conjugate 2. [Diagram 40] FIG. 40 is a TLC trace of 203Pb-conjugate 2. [Diagram 41] FIG. 41 is an HPLC trace of 203Pb-conjugate 2. [Diagram 42] FIG. 42 is a TLC trace of 203Pb-conjugate 2 after 40 minutes. [Diagram 43] FIG. 43 is a TLC trace of 203Pb-conjugate 2 after 2 hours. [Diagram 44] FIG. 44 is a TLC trace of 203Pb-conjugate 2 after 4 hours. [Diagram 45] FIG. 45 is a TLC trace of 203Pb-conjugate 2 after 21 hours. [Figure 46] FIG. 46 is an example of mixed labeling of conjugate 2 with 64Cu and 203Pb to form 64Cu / 203Pb-conjugate 2. [Figure 47] FIG. 47 is a TLC trace of 64Cu / 203Pb-conjugate 2 using a 0.15 M NH4Ac mobile phase. [Figure 48] FIG. 48 is a second TLC trace of 64Cu / 203Pb-conjugate 2 using a 0.1 M sodium citrate mobile phase. [Figure 49] Figure 49 is a TLC trace of 64Cu / 203Pb-conjugate 2 after 1 hour using two separate solvent systems. The first solvent system is 0.1 M sodium citrate. The second solvent system is 0.15 M NH4Ac. [Figure 50] Figure 50 is a TLC trace of 64Cu / 203Pb-conjugate 2 after 4 hours using two separate solvent systems. The first solvent system is 0.1 M sodium citrate. The second solvent system is 0.15 M NH4Ac. [Figure 51] Figure 51 is a TLC trace of 64Cu / 203Pb-conjugate 2 after 21 hours using two separate solvent systems. The first solvent system is 0.1 M sodium citrate. The second solvent system is 0.15 M NH4Ac. [Figure 52] FIG. 52 is an example of co-labeling of conjugate 2 with 64Cu and non-radioactive Pb to form 64Cu / Pb-conjugate 2. [Figure 53] FIG. 53 is a TLC trace of 64Cu / Pb-conjugate 2. [Figure 54] FIG. 54 is an HPLC trace of 64Cu / Pb-conjugate 2. [Figure 55] FIG. 55 is a graph of in vitro cellular uptake of 64Cu / Pb-conjugate 2 with or without Pb. [Figure 56] FIG. 56 includes various PET images of in vivo cellular uptake of 64Cu / Pb-conjugate 2 in mice at various time points after injection. [Figure 57] FIG. 57 is a graph of the SUV of 64Cu / Pb-conjugate 2 in organs of both normal and tumor-bearing mice. [Figure 58] FIG. 58 is a graph of the SUV of 64Cu / Pb-conjugate 2 with a molar specific activity of 0.325 GBq / μmol in mouse organs. [Figure 59] FIG. 59 is a graph of the SUV of 64Cu / Pb-conjugate 2 with a molar specific activity of 52 GBq / μmol in mouse organs. [Figure 60] FIG. 60 includes various PET images of the in vivo uptake of 64Cu / Pb-conjugate 2 in mice at various time points after injection. [Figure 61] FIG. 61 includes various PET images of the in vivo uptake of 64Cu / Pb-conjugate 2 in mice at various time points after injection. [Figure 62] FIG. 62 includes various graphs of SUV of 64Cu / Pb-conjugate 2 in tumors and kidneys of mice. [Figure 63] FIG. 63 includes various PET images of the in vivo uptake of 64Cu / Pb-conjugate 2 in mice at various time points after injection. [Figure 64] FIG. 64 includes various PET images of the in vivo uptake of 64Cu / Pb-conjugate 2 in mice at various time points after injection. [Figure 65] FIG. 65 includes various graphs of SUV of 64Cu / Pb-conjugate 2 in tumors and kidneys in mice. [Figure 66] FIG. 66 is a graph of in vitro cellular uptake of 64Cu-conjugate 2 with and without inhibitor. [Figure 67] FIG. 67 is a graph of the SUV of 64Cu-conjugate 2 in the kidney, tumor, and salivary gland of tumor-bearing mice 120 minutes after injection. [Figure 68] FIG. 68 is a graph of the SUV ratios of 64Cu-conjugate 2 in kidney vs. muscle, blood vs. muscle, tumor vs. muscle, and salivary gland vs. muscle in normal and tumor-bearing mice. [Figure 69]FIG. 69 includes various microPET images of the in vivo uptake of 64Cu-conjugate 2 in mice at various time points after injection. [Figure 70] FIG. 70 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and a NOTA chelator and a TCMC chelator. [Figure 71] FIG. 71 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and a NOTA chelator and a TCMC chelator. [Figure 72] FIG. 72 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and NOTA and MACROPA chelators. [Figure 73] FIG. 73 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and NOTA and MACROPA chelators. [Figure 74] FIG. 74 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and DFO and MACROPA chelators. [Figure 75] FIG. 75 is a representative example of the synthesis of a dual PSMA-targeting conjugate with alternative linker molecules and DFO and MACROPA chelators. [Figure 76] FIG. 76 is a representative example of the synthesis of a single PSMA-targeted conjugate with a NOTA chelator and a MACROPA chelator. [Figure 77] Figure 77 is a representative example of the synthesis of a single PSMA-targeted conjugate with a DFO chelator and a MACROPA chelator. [Figure 78] FIG. 78 is a representative example of the synthesis of a single FAP-targeted conjugate with a NOTA chelator and a MACROPA chelator. [Figure 79] FIG. 79 is a representative example of the synthesis of a single FAP-targeted conjugate with a DFO chelator and a MACROPA chelator. [Figure 80]FIG. 80 is a representative example of the synthesis of a single octreotide targeting conjugate with NOTA and MACROPA chelators. [Figure 81] FIG. 81 is a representative example of the synthesis of a single octreotide targeting conjugate with DFO and MACROPA chelators. [Figure 82] FIG. 82 is an example of labeling NOTA (Cu), TCMC (Pb) and FAPI conjugate (conjugate 3) with 64Cu to form 64Cu-conjugate 3. [Figure 83] FIG. 83 is an example of dual labeling of NOTA (Cu), TCMC (Pb) and FAPI conjugate (conjugate 3) with 64Cu and non-radioactive Pb to form 64Cu / Pb-conjugate 3. [Figure 84] FIG. 84 is a UV HPLC trace of 64Cu-conjugate 3. [Figure 85] Figure 85 is a rad-TLC trace of free [64Cu]CuCl2. [Figure 86] FIG. 86 is a rad-TLC trace of 64Cu-conjugate 3. [Figure 87] FIG. 87 is a rad-TLC trace of 64Cu / Pb-conjugate 3. [Figure 88] FIG. 88 is a UV HPLC trace of 64Cu / Pb-conjugate 3. [Figure 89] FIG. 89 is an emission HPLC trace of 64Cu / Pb-conjugate 3. [Figure 90] FIG. 90 is an example of dual labeling of NOTA (Cu), TCMC (Pb) and octreotide conjugate (conjugate 4) with 64Cu and non-radioactive Pb to form 64Cu / Pb-conjugate 4. [Figure 91] FIG. 91 is a UV HPLC trace of 64Cu / Pb-conjugate 4. [Figure 92] FIG. 92 is an emission HPLC trace of 64Cu / Pb-conjugate 4. [Figure 93] Figure 93 is a rad-TLC trace of free [64Cu]CuCl2. [Figure 94] FIG. 94 is a rad-TLC trace of 64Cu / Pb-conjugate 4. [Figure 95] FIG. 95 is an example of labeling conjugate 2 with 212Pb to form 212Pb-conjugate 2. [Figure 96] Figure 96 is a rad-TLC trace of [212Pb]PbCl2. [Figure 97] FIG. 97 is a rad-TLC trace of 212Pb-conjugate 2. [Figure 98] FIG. 98 is a rad-TLC trace of 212Pb-conjugate 2 2 hours after synthesis. [Figure 99] FIG. 99 is a rad-TLC trace of 212Pb-conjugate 2 22 hours after synthesis. [Figure 100] FIG. 100 is a series of images of a nude mouse bearing an LNCaP tumor before injection with 212Pb-conjugate 2 and an image of the nude mouse after injection with 212Pb-conjugate 2. [Figure 101] FIG. 101 is a series of PET images of a nude mouse bearing an LNCaP tumor before and after treatment with 212Pb-conjugate 2. [Figure 102] FIG. 102 is a representative example of a conjugate described herein having a cleavable linker. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0040] Detailed Description This document provides a conjugate comprising two or more chelators and one or more binding moieties or motifs, where one of the chelators is a chelator for an imaging isotope and one of the chelators is a chelator for a radiotherapeutic isotope. A trifunctional compound that can act as a linker (e.g., N',N'-bis(2-aminoethyl)ethane-1,2-diamine, etc.) can selectively react with two different chelators, one for an imaging isotope and one for a radiotherapeutic isotope, to generate a dual chelator compound. The dual chelator compound can be modified to make it suitable for reacting with a binding moiety, for example, at room temperature under mild reaction conditions (e.g., aqueous medium) to protect the nature and functionality of the binding moiety and generate a conjugate in which two or more chelators are covalently attached to one or more binding moieties or motifs. Only one functional group on the targeting binding moiety (e.g., primary NH2) is required to generate the conjugate. As described below, the combination of chelator and isotope can be varied according to the needs of the method of treatment or imaging.

[0041] In some embodiments, the chelator comprises a moiety of formula (I):

[0042] [ka] (In the formula: Each X is N, P, P(=O), CR N and a moiety of formula (i): [ka] are independently selected from; each of x1, x2, x3, and x4 independently represents a point of attachment of the moiety of formula (I) to a chelator or binding moiety; L 1 , L 2 , L 3 , and L 4 are C(=O), C(=S), and N(R N ), O, S, S(=O), S(=O)2, -CRN =NR N -, (-C 1-3 Alkylene-O-) x , (-OC 1-3 Alkylene-) x , -C 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, where each x is independently an integer from 1 to 10; 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 Each of arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from OH, NO, CN, halo, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Haloalkoxy, Amino, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 optionally substituted with 1, 2, or 3 substituents independently selected from alkoxycarbonyl; Each of y1, y2, y3, and y4 is independently an integer from 1 to 10; Each R N , H, C 1-3 Alkyl, and C 1-3 independently selected from haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5. can be attached to the binding moiety by

[0043] In some embodiments, X is N.

[0044] In some embodiments, X is P.

[0045] In some embodiments, X is P(=O).

[0046] In some embodiments, X is CR N It is.

[0047] In some embodiments, X is a moiety of formula (i).

[0048] In some embodiments, X is N or CR N is selected from.

[0049] In some embodiments, X is N, CR N and the moiety of formula (i).

[0050] In some embodiments, each L 1 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 arylene. In some embodiments, the moiety (L 1 ) y1 is represented by the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0051] In some embodiments, each L 2 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 arylene. In some embodiments, the moiety (L 2 ) y2 is represented by the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0052] In some embodiments, each L 3 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 arylene. In some embodiments, the moiety (L3 ) y3 is represented by the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0053] In some embodiments, each L 4 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 arylene. In some embodiments, the moiety (L 4 ) y4 is represented by the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0054] In some embodiments, y1 is an integer selected from 1, 2, 3, 4, 5, and 6. In some embodiments, y2 is an integer selected from 1, 2, 3, 4, 5, and 6. In some embodiments, y3 is an integer selected from 1, 2, 3, 4, 5, and 6. In some embodiments, y4 is an integer selected from 1, 2, 3, 4, 5, and 6.

[0055] In some embodiments, R N is H. In some embodiments, R N is C 1-3 In some embodiments, R N H and C 1-3 is selected from alkyl.

[0056] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0057] In some embodiments, the compound of formula (I) has the following formula:

[0058] [ka] has.

[0059] In some embodiments, the compound of formula (I) has the following formula:

[0060] [ka] has.

[0061] In some embodiments, the compound of formula (I) has the following formula:

[0062] [ka] has.

[0063] In some embodiments, the moiety of formula (I) has any one of the following formulas:

[0064] [ka] TIFF2024523344000014.tif146160.

[0065] In some embodiments, the moiety of formula (II):

[0066] [ka] (In the formula, x1 represents the point of attachment of formula (II) to the chelator; x2 represents the point of attachment of formula (II) to a chelator or binding moiety; Each L is C(=O), C(=S), N(R N ), O, S, S(=O), S(=O)2, -CR N =NR N -, (-C 1-3 Alkylene-O-) x , (-OC 1-3 Alkylene-) x , -C 1-3 Alkylene-, C 2-6Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, where each x is independently an integer from 1 to 10; 1-3 Alkylene-, C 2-6 Alkenylene, C 2-6 Alkynylene, C 3-10 Cycloalkylene, C 6-10 Each of arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from OH, NO, CN, halo, C 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy, C 1-3 Haloalkoxy, Amino, C 1-3 Alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 optionally substituted with 1, 2, or 3 substituents independently selected from alkoxycarbonyl; y is an integer from 1 to 30; Each R N , H, C 1-3 Alkyl, and C 1-3 haloalkyl) and a chelator is bound to the binding moiety and / or a chelator and a binding moiety are bound to the binding moiety.

[0067] In some embodiments, x2 represents the point of attachment of Formula (II) to the chelator. In some embodiments, x2 represents the point of attachment of Formula (II) to the binding moiety or a binding moiety.

[0068] In some embodiments, each L is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 In some embodiments, the moiety (L) is independently selected from arylene. y is represented by the formula NHC(=S)NH or C 6-10 Arylene-C 1-3Contains at least one alkylene- moiety.

[0069] In some embodiments, y is an integer from 1 to 10. In some embodiments, y is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, R N is H. In some embodiments, R N is C 1-3 In some embodiments, R N H and C 1-3 is selected from alkyl.

[0070] In some embodiments, the moiety of formula (II) has any one of the following formulae:

[0071] [ka] has.

[0072] In some embodiments, the chelator may be linked to the binding moiety with a cleavable linker. The term "cleavable linker" as used herein refers to a linker that is easily degraded or metabolized under certain conditions. In some examples, the cleavable linker may remain intact under most conditions (e.g., during storage), but may be cleaved when exposed to a particular compound (e.g., a compound present in the body, such as a particular protease), such that the linker is cleaved when in the presence of that compound. In some examples, the cleavable linker may remain intact under most conditions (e.g., during storage), but may be cleaved under physiological conditions (e.g., at natural human blood pH), such that the linker is cleaved when administered to a mammal (e.g., a human). For example, in some embodiments, the cleavable linker may be acid cleavable, GSH cleavable, Fe(II) cleavable, cathepsin cleavable, glycosidase cleavable, phosphatase cleavable, sulfatase cleavable, photoresponsive cleavable, or bioorthogonal cleavable. See, e.g., Zheng et al., Acta Pharm Sin B. 2021 Dec;11(12):3889-3907 and Tsusuchikama et al., Protein Cell. 2018 Jan;9(1):33-46. In some examples, the cleavable moiety can be as described in U.S. Pat. Nos. 11,191,854 or 10,093,741. For example, in some embodiments, the cleavable moiety can include an ester bond, a phosphate bond, or a disulfide bond. The ester bond can be cleavable by an esterase native to the cellular environment or can be hydrolyzable by a neutral or acidic buffered environment. The phosphate bond can be cleavable by a phosphatase or can be hydrolyzable by a neutral or acidic buffered environment. The disulfide bond can be cleavable by the reducing environment of the microenvironment, soluble GSH, thioredoxin, or glutaredoxin. Upon cleavage, the binding moiety can maintain its extended retention in the body, while the chelator and associated radionuclides can be rapidly excreted. One such schematic diagram for a conjugate described herein having cleavable ester bonds linking the Cu to the chelators for both Cu and Pb is shown in Figure 102. In Figure 102, the ester bonds can be replaced with phosphate or disulfide bonds.

[0073] In some embodiments, a cleavable linker can link a binding moiety to one or more chelators. For example, a cleavable linker can link a binding moiety to two chelators. In some embodiments, cleavage of the linker can separate one or more chelators from the binding moiety.

[0074] At various places in the present specification, substituents of compounds of the invention are disclosed in groups or in ranges. It is specifically intended that the invention include each and every individual subcombination of the members of such groups and ranges. For example, "C 1-6 The term "alkyl" is specifically intended to individually disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl.

[0075] Various aryl, heteroaryl, cycloalkyl, and heterocycloalkyl rings are described in various places in this specification. Unless otherwise specified, these rings can be attached to the rest of the molecule at any ring member, if allowed by valence. For example, the term "pyridine ring" or "pyridinyl" can refer to pyridin-2-yl, pyridin-3-yl, or pyridin-4-yl rings.

[0076] The term "aromatic" refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings having aromatic character (i.e., having (4n+2) delocalized π (pi) electrons, where n is an integer).

[0077] The term "n-membered," where n is an integer, typically describes the number of ring-forming atoms in a moiety where n is the number of ring-forming atoms. For example, piperidinyl is an example of a 6-membered heterocycloalkyl ring, pyrazolyl is an example of a 5-membered heteroaryl ring, pyridyl is an example of a 6-membered heteroaryl ring, and 1,2,3,4-tetrahydro-naphthalene is an example of a 10-membered cycloalkyl group.

[0078] As used herein, the expression "optionally substituted" means unsubstituted or substituted. The substituents are independently selected, and the substitution may occur at any chemically available position. As used herein, the term "substituted" means that a hydrogen atom has been removed and replaced with a substituent. A single divalent substituent, such as oxo, can replace two hydrogen atoms. It is understood that substitution at a given atom is limited by valence.

[0079] Throughout the definition, "C n-m " denotes a range inclusive of the endpoints, where n and m are integers and indicate the number of carbons. Examples include C 1-4 , C 1-6 etc.

[0080] As used herein, "C n-m The term "alkyl," used alone or in combination with other terms, refers to a saturated hydrocarbon group, which may be straight or branched, having n to m carbons. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, 1,2,2-trimethylpropyl, and the like. In some embodiments, an alkyl group contains 1 to 6 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms.

[0081] As used herein, "C n-mThe term "haloalkyl", used alone or in combination with other terms, refers to an alkyl group having from one halogen atom to 2s+1 halogen atoms, which may be the same or different (where "s" is the number of carbon atoms in the alkyl group), where the alkyl group has n to m carbon atoms. In some embodiments, the haloalkyl group is only fluorinated. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0082] As used herein, "C n-m "Alkenyl" refers to an alkyl group having one or more double carbon-carbon bonds and having n to m carbons. Examples of alkenyl groups include, but are not limited to, ethenyl, n-propenyl, isopropenyl, n-butenyl, sec-butenyl, and the like. In some embodiments, the alkenyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms.

[0083] As used herein, "C n-m "Alkynyl" refers to an alkyl group having one or more triple carbon-carbon bonds and having n to m carbons. Examples of alkynyl groups include, but are not limited to, ethynyl, propyn-1-yl, propyn-2-yl, and the like. In some embodiments, the alkynyl moiety contains 2 to 6, 2 to 4, or 2 to 3 carbon atoms.

[0084] As used herein, "C n-mThe term "alkylene", used alone or in combination with other terms, refers to a divalent alkyl linking group having n to m carbons. Examples of alkylene groups include, but are not limited to, ethane-1,1-diyl, ethane-1,2-diyl, propane-1,1,-diyl, propane-1,3-diyl, propane-1,2-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, 2-methyl-propane-1,3-diyl, and the like. In some embodiments, the alkylene moiety contains 2 to 6, 2 to 4, 2 to 3, 1 to 6, 1 to 4, or 1 to 2 carbon atoms. Similarly, "C n-m The term "alkenylene," alone or in combination with other terms, refers to a divalent alkenyl linking group having n to m carbons, and includes "C n-m The term "alkynyl," used alone or in combination with other terms, refers to a divalent alkynyl linking group having n to m carbons.

[0085] As used herein, "C n-m The term "alkoxy," used alone or in combination with other terms, refers to a group of formula -O-alkyl, where the alkyl group has n to m carbons. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (e.g., n-propoxy and isopropoxy), butoxy (e.g., n-butoxy and tert-butoxy), and the like. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0086] As used herein, "C n-m "Haloalkoxy" refers to a group of the formula -O-haloalkyl having n to m carbon atoms. An example of a haloalkoxy group is OCF3. In some embodiments, the haloalkoxy group is only fluorinated. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0087] As used herein, the term "amino" refers to a group of formula -NH2.

[0088] As used herein, "C n-m The term "alkylamino" refers to a group of formula -NH(alkyl), where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms. Examples of alkylamino groups include, but are not limited to, N-methylamino, N-ethylamino, N-propylamino (e.g., N-(n-propyl)amino and N-isopropylamino), N-butylamino (e.g., N-(n-butyl)amino and N-(tert-butyl)amino), and the like.

[0089] As used herein, "di(C n-m The term "-N(alkyl)amino" refers to a group of formula -N(alkyl)2, where the two alkyl groups each independently have n to m carbon atoms. In some embodiments, each alkyl group independently has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0090] As used herein, "C n-m The term "alkoxycarbonyl" refers to a group of formula -C(O)O-alkyl, where the alkyl group has n to m carbon atoms. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms. Examples of alkoxycarbonyl groups include, but are not limited to, methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl (e.g., n-propoxycarbonyl and isopropoxycarbonyl), butoxycarbonyl (e.g., n-butoxycarbonyl and tert-butoxycarbonyl), and the like.

[0091] The term "carboxy" as used herein refers to a -C(O)OH group. As used herein, "halo" refers to F, Cl, Br, or I. In some embodiments, halo is F, Cl, or Br.

[0092] As used herein, the term "aryl", used alone or in combination with other terms, refers to an aromatic hydrocarbon group that may be monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings). n-m The term "aryl" refers to an aryl group having n to m ring carbon atoms. Examples of aryl groups include phenyl, naphthyl, anthracenyl, phenanthrenyl, indanyl, indenyl, and the like. In some embodiments, an aryl group has 6 to 10 carbon atoms. In some embodiments, an aryl group is phenyl or naphthyl.

[0093] As used herein, "cycloalkyl" refers to a non-aromatic cyclic hydrocarbon, including cyclized alkyl and / or alkenyl groups. Cycloalkyl groups can include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) groups and spiro rings. The ring-forming carbon atoms of a cycloalkyl group can be optionally substituted with one or two independently selected oxo or sulfido groups (e.g., C(O) or C(S)). Also included within the definition of cycloalkyl are moieties having one or more aromatic rings fused to (i.e., sharing a bond with) the cycloalkyl ring, such as benzo or thienyl derivatives of cyclopentane, cyclohexane, and the like. Cycloalkyl groups containing fused aromatic rings can be bonded through any ring-forming atom, including the ring-forming atoms of the fused aromatic ring. Cycloalkyl groups can have 3, 4, 5, 6, 7, 8, 9, or 10 ring-forming carbons (C 3-10 In some embodiments, the cycloalkyl can have the formula: 3-10 In some embodiments, the cycloalkyl is a monocyclic or bicyclic cycloalkyl. 3-7Monocyclic cycloalkyl. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinyl, norcarnyl, adamantyl, and the like. In some embodiments, the cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0094] As used herein, "heteroaryl" refers to a monocyclic or polycyclic aromatic heterocycle having at least one heteroatom ring member selected from sulfur, oxygen, and nitrogen. In some embodiments, the heteroaryl ring has 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, any ring-forming N in the heteroaryl moiety can be an N-oxide. In some embodiments, the heteroaryl is a 5-10 membered monocyclic or bicyclic heteroaryl having 1, 2, 3, or 4 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl is a 5-6 monocyclic heteroaryl having 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur, and oxygen. In some embodiments, the heteroaryl is a 5- or 6-membered heteroaryl ring. A 5-membered heteroaryl ring is a heteroaryl having a ring with 5 ring atoms, where one or more (e.g., 1, 2, or 3) ring atoms are independently selected from N, O, and S. Exemplary 5-membered ring heteroaryls are thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, pyrazolyl, isothiazolyl, isoxazolyl, 1,2,3-triazolyl, tetrazolyl, 1,2,3-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-triazolyl, 1,2,4-thiadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-triazolyl, 1,3,4-thiadiazolyl, and 1,3,4-oxadiazolyl. A 6-membered ring heteroaryl is a heteroaryl having a ring with 6 ring atoms, where one or more (e.g., 1, 2, or 3) ring atoms are independently selected from N, O, and S. Exemplary 6-membered ring heteroaryls are pyridyl, pyrazinyl, pyrimidinyl, triazinyl, and pyridazinyl.

[0095] As used herein, "heterocycloalkyl" refers to a non-aromatic monocyclic or polycyclic heterocycle having one or more ring-forming heteroatoms selected from O, N, or S. Heterocycloalkyl includes monocyclic 4-, 5-, 6-, 7-, 8-, 9-, or 10-membered heterocycloalkyl groups. Heterocycloalkyl groups can also include spirocycles. Examples of heterocycloalkyl groups include pyrrolidin-2-one, 1,3-isoxazolidin-2-one, pyranyl, tetrahydropuran, oxetanyl, azetidinyl, morpholino, thiomorpholino, piperazinyl, tetrahydrofuranyl, tetrahydrothienyl, piperidinyl, pyrrolidinyl, isoxazolidinyl, isothiazolidinyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, imidazolidinyl, azepanyl, benzazapene, and the like. The ring-forming carbon atoms and heteroatoms of a heterocycloalkyl group may be optionally substituted with one or two independently selected oxo or sulfido groups (e.g., C(O), S(O), C(S), or S(O)2, etc.). A heterocycloalkyl group may be bonded through a ring-forming carbon atom or a ring-forming heteroatom. In some embodiments, a heterocycloalkyl group contains 0-3 double bonds. In some embodiments, a heterocycloalkyl group contains 0-2 double bonds. Similarly, the definition of heterocycloalkyl also includes moieties having one or more aromatic rings fused to (i.e., sharing a bond with) a cycloalkyl ring, such as benzo or thienyl derivatives of piperidine, morpholine, azepine, etc. Heterocycloalkyl groups containing fused aromatic rings may be bonded through any ring-forming atom, including the ring-forming atoms of the fused aromatic ring. In some embodiments, a heterocycloalkyl is a monocyclic 4-6 membered heterocycloalkyl having one or two heteroatoms independently selected from nitrogen, oxygen, or sulfur, and having one or more oxidized ring members. In some embodiments, the heterocycloalkyl is a mono- or bicyclic 4-10 membered heterocycloalkyl having 1, 2, 3, or 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur and having one or more oxidized ring members.

[0096] In certain places, definitions or embodiments refer to certain rings (e.g., azetidine rings, pyridine rings, etc.). Unless otherwise indicated, these rings may be attached to any ring member, provided that the valence of that atom is not exceeded. For example, an azetidine ring may be attached at any position on the ring, while a pyridin-3-yl ring is attached at the 3-position.

[0097] As used herein, the term "compound" is meant to encompass all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. Compounds identified herein as one particular tautomeric form by name or structure are intended to encompass other tautomeric forms unless otherwise specified.

[0098] The compounds described herein may be asymmetric (e.g., have one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the present invention that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods on how to prepare optically active forms from optically inactive starting materials are known in the art, such as by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, N=N double bonds, and the like, may also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and may be isolated as a mixture of isomers or as separated isomeric forms. In some embodiments, the compounds have the (R)-configuration. In some embodiments, the compounds have the (S)-configuration.

[0099] The compounds provided herein also encompass tautomeric forms. Tautomeric forms arise from the exchange of a single bond with an adjacent double bond and the concomitant migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and the same total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy more than one position of a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H- 1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms may be in equilibrium or sterically fixed in one form by appropriate substitution.

[0100] In some embodiments, each of the chelators can be independently, for example, NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, NTA, BisTris, EGTA, EDTA, BAPTA, DO2A, DO3A, and MACROPA. In general, the combination of chelators for imaging isotopes and therapeutic isotopes can be selected for a particular application. For example, in some embodiments, one chelator can be DiAmSar and one chelator can be TCMC. In some embodiments, one chelator can be NOTA and one chelator can be TCMC. In some embodiments, each of the chelators can be independently, a supermagnetic iron oxide nanoparticle (SPION). In some embodiments, the SPION can be ferumoxytol. Certain aspects of these embodiments are described, for example, in Advanced Drug Delivery Reviews, Vol. 63, No. 1-2, January-February 2011, pp. 24-46; and Kidney Int. 2017 Jul; 92(1): 47-66, which are incorporated by reference herein in their entireties.

[0101] In some embodiments, the conjugate may include three or more chelators. For example, in some embodiments, the conjugate may include three chelators, or four chelators, or five chelators. For example, in some embodiments, one chelator may be DiAmSar, one chelator may be TCMC, and one chelator may be NOTA. In some embodiments, each of the three chelators may be NOTA, or each of the above chelators may be SPION. In some embodiments, one chelator may be MACROPA, one chelator may be DFO, and one chelator may be DOTA. For example, in some embodiments, the conjugate may include three or more of DOTA, NOTA, TCMC, MACROPA, DiAmSar, and HBED. In some examples, one chelator may be DOTA, one chelator may be NOTA, one chelator may be TCMC, one chelator may be MACROPA, one chelator may be DiAmSar, and one chelator may be HBED.

[0102] The imaging and radiotherapeutic isotopes of the conjugates described herein can be selected to have similar half-lives. For example, the radiotherapeutic isotope can be an alpha emitter, e.g. 225 Ac, 212 Pb, 211 At, 213 Bi, 212 Bi, 211 Bi, 152 / 160 / 161 Tb, 227 Th, 223 Ra, 211 Po, 221 Fr, 217 At, 213 Po, 212 Po, 215 Po, or 177 Lu, and the imaging isotope may be 68 Ga, 44 Sc, 60 / 61 / 62 / 64 Cu, 84 / 86 / 87 / 89 Zr, 63 Zn, 43 / 44 Sc, 192 / 193 / 194 / 196 Au,52m Mn, 90 / 92m1 Nb, 51 / 52 Mn, 148 / 151 / 151m / 152 Tb, 45 Ti, 65 / 66 / 67 Ga, 94m Tc, 55 Co, 80 / 81 / 83 Sr, 38 K, 70 / 71 / 72 / 74 As, 81 / 82m Rb, 52 Fe, or 86 In some embodiments, the imaging isotope can be Y. 64 Cu, and the radiotherapy isotope is 212 It is Pb. 64 Cu is a positron-emitting PET imaging radionuclide that decays to a stable non-radioactive daughter nuclide 64 Ni and 64 It becomes Zn. 212 Pb is an alpha-emitting therapeutic radionuclide 212 The parent isotope of Bi, which eventually decays to a stable, non-radioactive daughter isotope 208 Pb. See, for example, Figure 1. 64 Cu has a physical half-life of 12.7 hours, 212 Pb has a physical half-life of 10.6 hours (or an effective physical half-life for alpha emission of 11.65 hours, as described below); 64 Using Cu as the imaging readout, 212 This makes it an ideal pair for assessing the biodistribution and dosimetry of Pb-related radioactivity. 68 Ga or 18 F) allows for a central production site covering most of the United States, and long-distance distribution of the resulting compounds. Strictly speaking, with regard to radioactive decay, 212 Pb decays into alpha emitters. 212 Bi, a beta-emitter. 212 Pb is 212 Because the beta emissions resulting from the decay of Pb are of little physiological importance compared to the alpha emissions, they are commonly referred to by physicians as alpha emitters. 212 After the Pb radionuclide emits beta radiation,212 Pb is 212 Bi (the daughter product) remains in the chelator and becomes part of the therapeutic agent. 212 Bi decays further by one of two equivalent pathways (see Figure 1); (1) 212 Bi emits alpha radiation 208 Tl and then emits beta radiation, or (2) 212 Bi emits beta radiation 212 It becomes Po, stays in the chelator, and then immediately emits alpha radiation. 212 Pb, and 212 Pb-containing drugs (including those described herein) were administered 11.65 hours ( 212 10.64 hours for Pb 212 Bi can be considered to be an alpha emitter with a physical half-life of 60.6 minutes (60.6 minutes for Bi). 212 The decay scheme of Pb (Figure 1) shows that it decays and becomes stable. 208 When it becomes Pb it gives one alpha emission and incidentally also gives two beta emissions. This has no significant effect as the beta emission is about 10,000 times less massive than the alpha emission, and therefore the two beta emissions are insignificant in terms of effects inside the body compared to the alpha emission. When beta emitters are used for therapy, the total amount of radiopharmaceutical that needs to be injected to have an effect is an order of magnitude higher than the dose of a comparable alpha emitter.

[0103] As shown in Figures 2A and 2B, depending on the desired application of the conjugate, different combinations of imaging and radiotherapeutic isotopes can be selected, resulting in conjugates that differ only in their radiation emission but have identical chemical structures and therefore identical binding affinities and biodistribution. For example, for non-radioactive conjugates, an inert radioactive metal isotope (e.g., 63 Cu and 208For imaging-only conjugates, imaging isotopes (e.g., Pb) can be selected for chelation with two or more chelators. 64 Cu) and inert radiotherapeutic isotopes (e.g. 208 For therapeutic-only conjugates, a radiotherapeutic isotope (e.g., 212 Pb) and inert imaging isotopes (e.g., 63 Cu) can be selected. In some embodiments, imaging-only and therapeutic-only conjugates can be prepared such that a desired dose (radioactively speaking) of each radioisotope is administered upon injection. For conjugates that can be used for simultaneous imaging and therapy, the imaging isotope (e.g., 64 Cu) and radiotherapeutic isotopes (e.g. 212 Pb) can be selected.

[0104] In some embodiments, a fluorescent dye is used instead of an imaging isotope. Non-limiting examples of fluorescent dyes include, for example, coumarins, cyanines, carboxyfluoresceins, quantum dots, green fluorescent protein (GFP), yellow fluorescent protein, red fluorescent protein, phycobiliproteins (e.g., phycoerythrin, phycocyanin, or allophycocyanin), xanthene derivatives, such as fluorescein or fluorescein isothiocyanate (fluorescein isothiocyanate), and the like. isthiocyanate) (FITC), rhodamine, Oregon Green, eosin, and Texas Red, cyanine derivatives such as cyanine, indocarbocyanine, oxacarbocyanine, thiacarbocyanine, and merocyanine; squaraine derivatives and ring-substituted squaraines such as the seta dyes and the squaraine dyes; squaraine rotaxane derivatives (e.g., tau dyes), naphthalene derivatives (e.g., dansyl derivatives and prodan derivatives); coumarin derivatives, oxadiazole derivatives (e.g., pyridyloxazole, nitrobenzoxadiazole, and benzoxadiazole); anthracene derivatives (e.g., anthraquinones, such as DRAQ5, DRAQ7, and CyTRAK Orange); pyrene derivatives (e.g., , Cascade Blue); oxazine derivatives (e.g., Nile Red, Nile Blue, Cresyl Violet, Oxazine 170); and acridine derivatives (e.g., Proflavine, Acridine Orange, Acridine Yellow); arylmethine derivatives (e.g., Auramine, Crystal Violet, Malachite Green); tetrapyrrole derivatives (e.g., porphine, phthalocyanine, bilirubin); dipyrromethane derivatives (e.g., BODIPY, aza-BODIPY); amino groups (active esters, carboxylates, isothiocyanates, hydrazines), carboxyl groups (carbodiimides), thiols (maleimides, acetyl bromides), or azides (via click chemistry or non-specifically (glutaraldehyde)).

[0105] For any conjugate, the binding moiety may be one or more small molecules, nanoparticles, liposomes, exosomes, polypeptides (e.g., antibodies or peptides), or any other targeting biologic that binds to a target molecule on a cell (e.g., a cancer cell). In some cases, the binding moiety may target a molecule on the surface of a cell (e.g., a cell surface receptor). For example, small molecules such as Glu-ureido-based prostate-specific membrane antigen (PSMA) inhibitors (also called glutamate carboxypeptidase II inhibitors) can be used as binding moieties. See, for example, Kopka, et al., J. Nucl. Med., 58(Suppl. 2):17S-26S (2017). PSMA (also called folate hydrolase 1 (FOLH1), FGCP, FOLH, GCP2, PSM, mGCP, GCPII, NAALAD1, or NAALA-dase) is a cell membrane peptidase that belongs to the M28B subfamily of the M28 peptidase family. For example, nanoparticles containing glutamate carboxypeptidase II inhibitors can be used for the binding moiety. In some embodiments, the nanoparticles can be hydrophilic polyethylene glycol coronas with small molecule PSMA targeting ligands. See, e.g., Autio, et al., JAMA Oncology, 4(10):1344-1351 (2018). Exosomes, such as dendritic cell-derived exosomes (see, e.g., Xu, et al., Molecular Cancer, 19, 160 (2020)), can be used for the binding moiety.

[0106] For example, in some embodiments, the binding moiety may be a polypeptide that binds to PSMA, a somatostatin receptor, a fibroblast activation protein (FAP) polypeptide, a melanocortin-1 receptor, a B7-H3 protein, a CA19-9 expressing tumor, cluster of differentiation 37 (CD37), cluster of differentiation 3 (CD3), cluster of differentiation 20 (CD20), cxc-motif chemokine receptor 4 (CXCR4), a gastrin releasing peptide receptor (GRPR), human epidermal growth factor receptor 2 (HER2), melanocortin-1 receptor (MC1R), somatostatin receptor 2 (SSTR2), vascular endothelial growth factor (VEGF), a programmed death-ligand 1 (PD-L1) polypeptide, a tumor-associated calcium signal transducer 2 (TROP2) polypeptide, a protein tyrosine kinase 2 (PTK2) polypeptide, an integrin beta 6 (ITGB6) polypeptide, a neurotensin receptor ligand, CD8, or vitamin B-12. See, e.g., Langbein et al., J. Nucl. Med., 60(Suppl. 2):13S-19S (2019). For example, the polypeptide can be a somatostatin analog such as Phe1-Tyr3-octreotate (TATE) or Phe1-Tyr3-octreotide (TOC). See, e.g., Stueven et al., Int. J. Mol. Sci., 20(12):3049 (2019). In some embodiments, the conjugate comprises two different polypeptides. In some embodiments, the polypeptide can be an antibody or an antibody fragment having the ability to bind to an antigen. The term "antibody" as used herein encompasses monoclonal antibodies, polyclonal antibodies, recombinant antibodies, humanized antibodies, chimeric antibodies, nanobodies, or multispecific antibodies (e.g., bispecific antibodies) formed from at least two antibodies. The term "antibody fragment" includes any portion of the above-mentioned antibodies, such as their antigen-binding region or variable region (e.g., a single VH domain). The term "epitope" refers to an antigenic determinant on an antigen to which the paratope of an antibody binds.Epitopic determinants usually consist of chemically active surface groupings of molecules (eg, amino acid residues or sugar residues) and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.

[0107] Examples of antibody fragments include Fab fragments, Fab' fragments, F(ab')2 fragments, Fv fragments, diabodies, single-chain antibody molecules, single VH domains, and other fragments thereof, so long as the fragments exhibit the desired binding ability to the target molecule. An "Fv fragment" is the smallest antibody fragment that contains an intact antigen recognition and binding site. This region consists of a dimer of one heavy chain variable domain and one light chain variable domain in tight non-covalent association. In this configuration, the three complementarity determining regions (CDRs) of each variable domain interact to define an antigen binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen binding specificity to the antibody. However, even a single variable domain (or half of an Fv containing only three CDRs specific for an antigen) has the ability to recognize and bind to an antigen, although usually with a lower affinity than the intact binding site. A "Fab fragment" is a fragment of an antibody that contains the constant domain of the light chain and the first constant domain of the heavy chain (CDR1, CDR2, CDR3, CDR4, CDR5, CDR6, CDR7, CDR8, CDR9, CDR10, CDR11, CDR12, CDR13, CDR14, CDR15, CDR16, CDR17, CDR18, CDR19, CDR10, CDR11, CDR12, CDR15, CDR16, CDR17, CDR18, CDR19, CDR11, CDR12, CDR13, CDR14, CDR15, CDR16, CDR17, CDR18, CDR19 ...9, CDR12, CDR13, CDR14, CDR15, CDR16, H1 "Fab fragments" also include the heavy chain C fragments, including one or more cysteines from the antibody hinge region. H1 It differs from an "Fab' fragment" by the addition of a few residues at the carboxy terminus of the domain. An "F(ab')2 fragment" is initially produced as a pair of "Fab' fragments" which have hinge cysteines between them. Methods for producing such antibody fragments, for example papain or pepsin digestion, can be performed using any suitable method.

[0108] In some cases, the antibody may be a humanized monoclonal antibody. Humanized monoclonal antibodies can be produced by introducing mouse complementarity determining regions (CDRs) from the heavy and light variable chains of mouse immunoglobulins into a human variable domain, followed by substituting human residues in the framework regions of the mouse counterpart. The use of antibody components derived from humanized monoclonal antibodies eliminates potential problems associated with the immunogenicity of mouse constant regions when treating humans. General techniques for cloning mouse immunoglobulin variable domains are described, for example, by Orlandi et al., Proc. Nat'l. Acad. Sci. USA 86:3833 (1989). Techniques for producing humanized monoclonal antibodies have been described, for example, by Jones et al., Nature 321:522 (1986); Riechmann et al., Nature 332:323 (1988); Verhoeyen et al., Science 239:1534 (1988); Carter et al., Proc. Nat'l. Acad. Sci. USA 89:4285 (1992); and Sandhu, Crit. Rev. Biotech. 12:437 (1992); Singer et al., J. Immunol. 150:2844 (1993). In some cases, humanization, e.g., hyperhumanization, can be used as described in Hwang et al., Methods, 36:35-42 (2005).In some cases, CDR grafting (Kashmiri et al., Methods, 36:25-34 (2005)), human string content optimization (Lazar et al., Mol. Immunol., 44:1986-1998 (2007)), framework shuffling (Dall'Acqua et al., Methods, , 36:43-60 (2005); and Damschroder et al., Mol. Immunol., 44:3049-3060 (2007)), and phage display approaches (Rosok et al., J. Biol. Chem., 271:22611-22618 (1996); Radar et al., Proc. Natl Acad. Sci. USA, 95:8910-8915 (1998); and Huse et al., Science, 246:1275-1281 (1989)) can be used to obtain antibody preparations that bind to the target molecule. In some cases, fully human antibodies can be generated from recombinant human antibody library screening techniques, for example, as described by Griffiths et al., EMBO J., 13:3245-3260 (1994); and Knappik et al., J. Mol. Biol., 296:57-86 (2000).

[0109] Antibody fragments can be prepared by proteolytic hydrolysis of an intact antibody or by expression of a nucleic acid encoding the fragment. Antibody fragments can be obtained by pepsin or papain digestion of an intact antibody by conventional methods. For example, Fab fragments can be produced by enzymatic cleavage of an antibody with papain. In some examples, antibody fragments can be produced by enzymatic cleavage of an antibody with pepsin to provide a 5S fragment designated F(ab')2. This fragment can be further cleaved using a thiol reducing agent and, optionally, a blocking group for the sulfhydryl groups resulting from cleavage of disulfide bonds to produce a 3.5S Fab' monovalent fragment. In some examples, enzymatic cleavage with pepsin can be used to directly produce two monovalent Fab' fragments and an Fc fragment. These methods are described, for example, by Goldenberg (U.S. Patent Nos. 4,036,945 and 4,331,647). See also Nisonhoff et al., Arch. Biochem. Biophys. 89:230 (1960); Porter, Biochem. J. 73:119 (1959); Edelman et al., METHODS IN ENZYMOLOGY, Vol. 1, p. 422 (Academic Press 1967); and Coligan et al., Sections 2.8.1 2.8.10 and 2.10.1 2.10.4.

[0110] The antibody may be an IgA, IgD, IgE, IgG or IgM type antibody, such as an IgG or IgM type antibody, including, but not limited to, an IgG1, IgG2, IgG3, IgG4, IgM1 and IgM2 type antibody. For example, in some examples, the antibody is an IgG1, IgG2 or IgG4 type antibody.

[0111] In some embodiments, the antibody may be an antibody that binds to PSMA. For example, an antibody that binds to PSMA may comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequence set forth in SEQ ID NO: 1-6. In some examples, an antibody that binds to PSMA may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NO: 1-6, provided that the antigen-binding domain retains the ability to bind to PSMA. For example, one or more CDRs of an antibody that binds to PSMA may consist of the amino acid sequence set forth in any one of SEQ ID NO: 1-6, except that the variant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NO: 1-6), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NO: 1-6), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NO: 1-6), provided that the antibody retains the ability to bind to PSMA. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 1-6 and that can be used in antibodies that bind to PSMA include, but are not limited to, the amino acid sequences set forth in Table 1 (see also Example 17).

[0112] [Table 1] TIFF2024523344000018.tif234159TIFF2024523344000019.tif163158

[0113] In some embodiments, the antibody that binds to PSMA can be as described elsewhere, e.g., in U.S. Pat. No. 10,179,819, International Patent Application Publication No. WO 2018 / 129284, International Patent Application Publication No. WO 2002 / 098897, U.S. Pat. Appl. Pub. No. 2014 / 0273078, European Patent Application Publication No. 3192810 A1, CN 108699157, European Patent No. 2,363,404, U.S. Pat. Appl. Pub. No. 2014 / 0234215, International Patent Application Publication No. WO 2005 / 094882, U.S. Pat. No. 7,666,414, U.S. Pat. No. 8,114,965, U.S. Pat. No. 8,470,330, International Patent Application Publication No. WO See U.S. Patent No. 2014 / 4057113, U.S. Patent No. 9,242,012, U.S. Patent No. 10,179,819, and U.S. Patent No. 9,782,478.

[0114] In some embodiments, the antibody that binds to PSMA can be a J591 monoclonal antibody or a humanized J591 monoclonal antibody. See, e.g., Milowsky et al., J. Nucl. Med., 50:606-11 (2009). Fully human monoclonal antibodies that bind to PSMA can also be used. See, e.g., Ma et al., Clin. Cancer Res., 12(8):2591-6 (2006).

[0115] In some embodiments, the antibody may be an antibody that binds to a somatostatin receptor polypeptide. Examples of somatostatin receptor polypeptides include, but are not limited to, sstr1 receptor polypeptide, sstr2a receptor polypeptide, sstr2b receptor polypeptide, sstr3 receptor polypeptide, sstr4 receptor polypeptide, and sstr5 receptor polypeptide. For example, an antibody that binds to a somatostatin receptor may comprise a CDR that comprises, consists essentially of, or consists of a CDR amino acid sequence as set forth in SEQ ID NOs: 7-12. In some examples, an antibody that binds to a somatostatin receptor provided herein may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 7-12, provided that the antigen binding domain retains the ability to bind to a somatostatin receptor. For example, one or more CDRs of the antibodies that bind to somatostatin receptors provided herein may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 7-12, except that the variant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 7-12), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 7-12), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 7-12), provided that the antigen binding domain retains the ability to bind to somatostatin receptors. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 7-12 and that may be used in antibodies that bind to somatostatin receptors include, but are not limited to, the amino acid sequences set forth in Table 2 (see also Example 17).

[0116] [Table 2]

[0117] In some embodiments, the antibody that binds to a somatostatin receptor may be UMB1, UMB4, UMB5, or UMB7.

[0118] In some embodiments, the antibody that binds to the somatostatin receptor can be as described elsewhere.See, for example, International Patent Application Publication No. WO 2018 / 005706, US Patent Application Publication No. 2009 / 0016989, US Patent Application Publication No. 2021 / 0340264, US Patent No. 11,225,521, NZ 749841A, AU 2017290086A, CN 201780041351.9A, and Korner et al., Am J Surg Pathol. 2012 Feb;36(2):242-52.

[0119] In some embodiments, the antibody may be an antibody that binds to a FAP polypeptide. For example, an antibody that binds to a FAP polypeptide may comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequences set forth in SEQ ID NOs: 13-18. In some examples, an antibody that binds to a FAP polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 13-18, provided that the antigen binding domain retains the ability to bind to a FAP polypeptide. For example, one or more CDRs of an antibody that binds a FAP polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 13-18, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 13-18), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 13-18), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 13-18), provided that the antigen-binding domain retains the ability to bind to a FAP polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 13-18 and that may be used in antibodies that bind to a FAP polypeptide include, but are not limited to, the amino acid sequences set forth in Table 3 (see also Example 17).

[0120] [Table 3] TIFF2024523344000022.tif50159

[0121] In some embodiments, an antibody that binds to a FAP polypeptide can be sibrotuzumab or BMS168.

[0122] In some embodiments, an antibody that binds to a FAP polypeptide can be as described elsewhere, see, e.g., JP 7017599 B2, JP 2009522329 A, US Patent Application Publication No. 2021 / 0253736, EP 3269740 A1, US Patent Application Publication No. 8,999,342, US Patent Application Publication No. 2017 / 0369592, IL 281739 D0, US Patent No. 9,481,730, and ES 2348556 T3.

[0123] In some embodiments, the antibody may be an antibody that binds to a CD3 polypeptide. For example, an antibody that binds to a CD3 polypeptide may comprise a CDR that comprises, consists essentially of, or consists of a CDR amino acid sequence set forth in SEQ ID NOs: 19-24. In some examples, an antibody that binds to a CD3 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 19-24, provided that the antigen binding domain retains the ability to bind to a CD3 polypeptide. For example, one or more CDRs of an antibody that binds to a CD3 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 19-24, except that the variant polypeptide comprises 1, 2, 3, 4 or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 19-24), has 1, 2, 3, 4 or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 19-24), and / or has 1, 2, 3, 4 or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 19-24), provided that the antigen-binding domain retains the ability to bind to a CD3 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 19-24 and that may be used in an antibody that binds to a CD3 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 4 (see also Example 17).

[0124] [Table 4]

[0125] In some embodiments, the antibody that binds to a CD3 polypeptide can be muromonab or blinatumomab.

[0126] In some embodiments, an antibody that binds a CD3 polypeptide can be as described elsewhere, see, e.g., CN 1984931 A, EP 1753783 B1, AU 2009 / 299792 B2, CN 102796199 A, and JP 6817211 B2.

[0127] In some embodiments, the antibody may be an antibody that binds to a CD20 polypeptide. For example, an antibody that binds to a CD20 polypeptide may comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequence set forth in SEQ ID NOs: 25-30. In some examples, an antibody that binds to a CD20 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 25-30, provided that the antigen binding domain retains the ability to bind to a CD20 polypeptide. For example, one or more CDRs of an antibody that binds to a CD20 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 25-30, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 25-30), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 25-30), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 25-30), provided that the antigen-binding domain retains the ability to bind to a CD20 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 25-30 and that may be used in antibodies that bind to a CD20 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 5 (see also Example 17).

[0128] [Table 5] TIFF2024523344000025.tif20159

[0129] In some embodiments, an antibody that binds to a CD20 polypeptide can be tositumomab, tituximab, ofatumumab, obinutuzumab, ocrelizumab, or ublituximab.

[0130] In some embodiments, an antibody that binds a CD20 polypeptide can be as described elsewhere, see, e.g., EP 1740946 B1, U.S. Patent No. 8,147,832, EP 1692182 B1, EP 2295468 B1, U.S. Patent Application Publication No. 2004 / 0093621 A1, U.S. Patent No. 7,744,877, CN 1210307 C, and CN 104558191 A.

[0131] In some embodiments, the antibody may be an antibody that binds to a CXCR4 polypeptide. For example, an antibody that binds to a CXCR4 polypeptide may comprise a CDR that comprises, consists essentially of, or consists of a CDR amino acid sequence set forth in SEQ ID NOs: 31-36. In some examples, an antibody that binds to a CXCR4 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 31-36, provided that the antigen binding domain retains the ability to bind to a CXCR4 polypeptide. For example, one or more CDRs of an antibody that binds to a CXCR4 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 31-36, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 31-36), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 31-36), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 31-36), provided that the antigen-binding domain retains the ability to bind to a CXCR4 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 31-36 and that may be used in an antibody that binds to a CXCR4 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 6 (see also Example 17).

[0132] [Table 6] TIFF2024523344000027.tif228160TIFF2024523344000028.tif118160

[0133] In some embodiments, the antibody that binds to a CXCR4 polypeptide can be ibalizumab, MAB172-100, PA3-305, or hz515H7.

[0134] In some embodiments, an antibody that binds to a CXCR4 polypeptide can be as described elsewhere. See, for example, EP 2285833 B1, JP 5749330 B2, U.S. Pat. No. 7,138,496, U.S. Pat. Appl. Pub. No. 2005 / 0002939, EP 2246364 A1, CA 2724409 A1, International Pat. Appl. Pub. No. WO 2006 / 089141, Broussas et al., Mol. Cancer Ther., 2016 Aug; 15(8):1890-9, International Pat. Appl. Pub. No. WO 2000 / 042074, International Pat. Appl. Pub. No. WO 2004 / 059285, EP 1449850 A1, TW I469792 B, U.S. Pat. No. 8,329,178, U.S. Pat. No. 7,892,546, International Pat. Appl. Pub. No. WO See, for example, US Patent Publication No. 2009 / 138519, International Patent Application Publication No. WO 2009 / 140124, International Patent Application Publication No. WO 2008 / 142303, International Patent Application Publication No. WO 2008 / 060367, U.S. Patent No. 8,748,107, TW I469792 B, RU 2636032 C2, U.S. Patent No. 10,428,151, CN 106211774 B, EP 1871807 B1, U.S. Patent Application Publication No. 2019 / 0276544, EP 06748215 A, U.S. Patent No. 8,329,178, and CA 2597717 A.

[0135] In some embodiments, the antibody may be an antibody that binds to a GRPR polypeptide.

[0136] In some embodiments, an antibody that binds to a GRPR polypeptide can be ABR-002, sc-398549, A30653.

[0137] In some embodiments, an antibody that binds a GRPR polypeptide may be as described elsewhere, see, e.g., CA 2089212 C, DE 69637411 T2, EP 0981369 B1, CN 109422810 A, CN 106132993 A, and International Patent Application Publication No. WO 2015 / 143525.

[0138] In some embodiments, the antibody may be an antibody that binds to a HER2 polypeptide. For example, an antibody that binds to a HER2 polypeptide may comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequence set forth in SEQ ID NOs: 37-42. In some examples, an antibody that binds to a HER2 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 37-42, provided that the antigen binding domain retains the ability to bind to a HER2 polypeptide. For example, one or more CDRs of an antibody that binds to a HER2 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 37-42, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 37-42), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 37-42), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 37-42), provided that the antigen-binding domain retains the ability to bind to a HER2 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 37-42 and that may be used in an antibody that binds to a HER2 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 7 (see also Example 17).

[0139] [Table 7]

[0140] In some embodiments, the antibody that binds to a HER2 polypeptide can be trastuzumab, pertuzumab, margetuximab, ZW25, or zumuzumab.

[0141] In some embodiments, the antibody that binds to the HER2 polypeptide can be as described elsewhere.See, for example, Jones et al., Nature, 321, 522-525 (1986), CN 105829346 B, CN 107001479 B, KR 2014 / 0032004 A, AU 2005 / 32520, TW I472339 B, CN 102167742 B, ES 2640449 T3, KR 20170055521 A, CN 111741979 A, ​​International Patent Application Publication No. WO 2021 / 097220, and CN 107001479 B.

[0142] In some embodiments, the antibody can be an antibody that binds to an MCR1 polypeptide.

[0143] In some embodiments, an antibody that binds to an MCR1 polypeptide may be ARC0638 or EPR6530.

[0144] In some embodiments, the antibody may be an antibody that binds to a VEGF polypeptide. Examples of VEGF polypeptides include VEGF1, VEGFB, VEGFC, and VEGFD. For example, an antibody that binds to a VEGF polypeptide may comprise a CDR that comprises, consists essentially of, or consists of a CDR amino acid sequence set forth in SEQ ID NOs: 43-48. In some examples, an antibody that binds to a VEGF polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 43-48, provided that the antigen binding domain retains the ability to bind to a VEGF polypeptide. For example, one or more CDRs of an antibody that binds a VEGF polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 43-48, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 43-48), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 43-48), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 43-48), provided that the antigen-binding domain retains the ability to bind to an MCR1 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 43-48 and that may be used in an antibody that binds a VEGF polypeptide include, but are not limited to, the amino acid sequences set forth in Table 8 (see also Example 17).

[0145] [Table 8] TIFF2024523344000031.tif145161

[0146] In some embodiments, the antibody that binds to a VEGF polypeptide can be bevacizumab, ranibizumab, brolucizumab, or faricimab.

[0147] In some embodiments, the antibody may be an antibody that binds to a PD-L1 polypeptide. For example, an antibody that binds to a PD-L1 polypeptide may comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequence set forth in SEQ ID NOs: 345-350. In some examples, an antibody that binds to a PD-L1 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) to the CDRs set forth in any one of SEQ ID NOs: 345-350, provided that the antigen binding domain retains the ability to bind to a PD-L1 polypeptide. For example, one or more CDRs of an antibody that binds to a PD-L1 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 345-350, except that the variant polypeptide comprises one, two, three, four or five amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 345-350), has one, two, three, four or five amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 345-350), and / or has one, two, three, four or five amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 345-350), provided that the antigen-binding domain retains the ability to bind to a PD-L1 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 345-350 and that may be used in antibodies that bind to a PD-L1 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 9 (see also Example 17).

[0148] [Table 9] TIFF2024523344000033.tif24160

[0149] In some embodiments, the antibody that binds to a PD-L1 polypeptide may be atezolizumab, avelumab, durvalumab, BMS 936559, or cosibelimab.

[0150] In some embodiments, the antibody may be an antibody that binds to a TROP2 polypeptide. For example, an antibody that binds to a VEGF polypeptide may comprise a CDR that comprises, consists essentially of, or consists of a CDR amino acid sequence as set forth in SEQ ID NOs: 351-356. In some examples, an antibody that binds to a TROP2 polypeptide may have one or more CDRs that are variants (e.g., not 100% identical) of the CDRs set forth in any one of SEQ ID NOs: 351-356, provided that the antigen-binding domain retains the ability to bind to a TROP2 polypeptide. For example, one or more CDRs of an antibody that binds to a TROP2 polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 351-356, except that the mutant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions in the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 351-356), has 1, 2, 3, 4, or 5 amino acid residues before the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 351-356), and / or has 1, 2, 3, 4, or 5 amino acid residues after the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 351-356), provided that the antigen-binding domain retains the ability to bind to a TROP2 polypeptide. Examples of CDR amino acid sequences that comprise, consist essentially of, or consist of the CDR amino acid sequences set forth in SEQ ID NOs: 351-356 and that may be used in an antibody that binds to a TROP2 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 10 (see also Example 17).

[0151] [Table 10] TIFF2024523344000035.tif82161

[0152] In some embodiments, the antibody that binds to a TROP2 polypeptide may be sacituzumab or datopotamab.

[0153] In some embodiments, the conjugate can be prepared as shown in any one or more of Figures 3-9 and 70-81. For example, in some embodiments, one chelator can be DiAmSar and one chelator can be TCMC. In some embodiments, one chelator can be NOTA and one chelator can be TCMC. In some embodiments, the binding moiety is a PSMA peptide. In some embodiments, the PSMA peptide is piflufostat. In some embodiments, the binding moiety is fibroblast activation protein inhibitor (FAPI). In some embodiments, the FAPI is N-[2-[(2S)-2-cyano-4,4-difluoropyrrolidin-1-yl]-2-oxoethyl]-6-hydroxyquinoline-4-carboxamide. In some embodiments, the binding moiety is a ligand for the somatostatin receptor. In some embodiments, the ligand for the somatostatin receptor is octreotide, pasireotide, vapreotide, lanreotide, somatostatin, edotreotide, or oxodotreotide. In some embodiments, the binding moiety is a CD3 inhibitor. In some embodiments, the binding moiety is a CD20 inhibitor. In some embodiments, the binding moiety is a CXCR4 inhibitor. In some embodiments, the CXCR4 inhibitor is framycetin, plerixafor, baclofen, mavorixafor, or MSX-122. In some embodiments, the binding moiety is a GRPR inhibitor. In some embodiments, the GRPR inhibitor is bombesin, RC-3095, PD 168368, GRPR antagonist 1, GRPR antagonist 2, or PD 176252. Some examples of GRPR antagonists that can be used as described herein are shown in Yu et al., Med Chem Res 30, 2069-2089 (2021), which is incorporated herein by reference. In some embodiments, the binding moiety is a HER2 inhibitor.In some embodiments, the HER2 inhibitor is lapatinib, tesevatinib, varlitinib, tucatinib, afatinib, brigatinib, fostamatinib, zanubrutinib, tucatinib, or neratinib. In some embodiments, the binding moiety is an MC1R ligand. In some embodiments, the MC1R ligand is 4-phenylbutyryl-His-DPhe-Arg-Trp-Gly-Lys(hex-5-ynoyl)-NH2, H-Lys(hex-5-ynoyl)-Tyr-Val-Nle-Gly-His-DNal(2')-Arg-DTrp-Asp-Arg-Phe-Gly-NH2, H-Lys(hex-5-ynoyl)Tyr-Val-Nle-Gly-His-DNal(2')-Arg-DPhe-Asp-Arg-Phe-Gly-NH2, adrenocorticotropic hormone, alpha melanocyte stimulating hormone, beta melanocyte stimulating hormone, gamma melanocyte stimulating hormone, or MC1RL. Some further examples of MC1R ligands that may be used as described herein are as set forth in one or more of the following: Tafreshi et al., J. Nucl. Med. 60(8), 1124-1133 (2019); and U.S. Patent Nos. 8,492,517, 8,933,194, and 11,286,280, which are incorporated herein by reference. In some embodiments, the binding moiety is a VEGF inhibitor.In some embodiments, the VEGF inhibitor is sunitinib, vatalanib, linifanib, denibulin, pazopanib, axitinib, regorafenib, sorafenib, lenvatinib, nintedanib, polaprezinc, fostamatinib, selpercatinib, or tivozanib. In some embodiments, the binding moiety is a PD-L1 inhibitor. In some embodiments, the PD-L1 inhibitor is AUNP-12, CA-170, (3S,3aR,6S,6aR)-N6-[4-(3-fluorophenyl)-pyrimidin-2-yl]-N3-(2-pyridylmethyl)-2,3,3a,5,6,6a-hexahydrofu, or 1-isopropyl-3-[(3S,5S)-1-methyl-5-[3-(2-naphthyl)-1,2,4-oxadiazol-5-yl]pyrrolidin-3-yl]urea. In some embodiments, the binding moiety is a PTK2 inhibitor. In some embodiments, the PTK2 inhibitor is endostatin, fostamatinib, 7-pyridin-2-yl-N-(3,4,5-trimethoxyphenyl)-7h-pyrrolo[2,3-D]pyrimidin-2-amine, 2-({5-chloro-2-[(2-methoxy-4-morpholin-4-ylphenyl)amino]pyrimidin-4-yl}amino)-N-methylbenzamide, GSK2256098, defactinib, or VS-4718. In some embodiments, the binding moiety is an ITGB6 binding agent. In some embodiments, the ITGB6 binding agent is a cyclic peptide cyclo(FRGDLAFp(NMe)K) or tribehexin, as described in Quigley et al., Eur J. Nucl. Med. Mol. Imaging. 49(4), 1136-1147 (2022).The binding moiety may be 3-fluoro-2,2-dimethylpropionic acid or 2,2-dimethylpropionic acid.

[0154] As described herein, the conjugates provided herein may include one or more binding moieties (e.g., 1, 2, 3, 4, 5 or more binding moieties). In some examples, the binding moieties of the conjugates described herein may be capable of binding to one or more target molecules. For example, the binding moieties of the conjugates described herein may be capable of binding to 1, 2, 3, 4, 5 or more target molecules, for example, 1, 2, 3, 4, 5 or more target molecules present on a cell (e.g., a cancer cell).

[0155] In some embodiments, conjugates provided herein having two or more binding moieties can advantageously bind, for example, to antigens present on two different cells (e.g., two different cancer cells) or to two different antigens on the same cell (e.g., the same cancer cell). In some embodiments, having two or more binding moieties provides the conjugate with one or more advantages, for example, enhanced uptake and / or increased in vivo stability.

[0156] In some embodiments, one or more of the conjugates described herein can be used to treat cancer (e.g., prostate cancer, neuroendocrine cancer, colon cancer, lung cancer, pancreatic cancer, melanoma, or lymphatic cancer) in a mammal (e.g., a human patient). For example, to treat prostate cancer, a conjugate comprising a binding moiety that targets PSMA or its activity can be used. To treat neuroendocrine cancer, a conjugate comprising a binding moiety that targets a somatostatin receptor (e.g., a somatostatin analog) can be used. To treat lung cancer, a conjugate comprising a binding moiety that targets a B7-H3 protein can be used. To treat pancreatic cancer, a conjugate comprising a binding moiety that targets a C9-19 can be used. To treat melanoma, a conjugate comprising a binding moiety that targets a melanocortin-1 receptor can be used.

[0157] In some embodiments, one or more of the conjugates described herein can be used to treat a non-cancerous condition (e.g., a benign tumor, an inflammatory condition, a hematologic process, a histiocytic process, a cystic disease, or an infectious disease) in a mammal (e.g., a human patient).

[0158] In some embodiments, one or more conjugates described herein can be administered to a mammal (e.g., a human patient) one or more times over a period of days to months to treat a cancerous or non-cancerous condition in the mammal (e.g., a human patient). In some embodiments, one or more conjugates described herein (e.g., a conjugate comprising two or more chelators covalently attached via a linker to a binding moiety, one of the chelators being a chelator of an isotope used for imaging and one of the chelators being a chelator of an isotope used for radiotherapy, where the isotope used for imaging and the isotope used for radiotherapy are each complexed (chelated) to the chelator, and where the binding moiety binds to a tumor in the patient) can be formulated into a pharma- ceutically acceptable composition for administration to a patient (e.g., a patient identified as having cancer) to treat the cancer in the patient. In some embodiments, a mixture of two conjugates can be administered, for example, to provide a suitable dose (radioactively speaking) of each radioisotope upon injection. In such an embodiment, the appropriate isotopes can be complexed with the chelators of the two conjugates and mixed at the time of injection, accounting for their decay at different rates.

[0159] In some embodiments, a conjugate comprising two or more chelators covalently attached to a binding moiety via a linker, one of the chelators being a chelator for an imaging isotope and one of the chelators being a chelator for a radiotherapeutic isotope, wherein the imaging isotope is chelated to the chelator, and wherein the binding moiety is bound to a tumor in a patient, can be administered to determine the biodistribution of the conjugate in the patient (i.e., the location of the conjugate in the mammal) (e.g., by PET). After the biodistribution is determined, the patient can be administered a conjugate that is identical except that both the imaging isotope and the radiotherapeutic isotope are chelated to the chelator. Determining the biodistribution allows the dose of treatment to be personalized to the patient, reducing side effects. Imaging can be performed after each administration of the conjugate to monitor the treatment.

[0160] The therapeutically effective amount of the conjugates described herein can be formulated with one or more pharma- ceutically acceptable carriers (additives or excipients) and / or diluents. In some embodiments, the additives stabilize against radiolysis. The pharmaceutical compositions can be formulated for administration in solid or liquid form, including, but not limited to, sterile solutions, suspensions, sustained release formulations, tablets, capsules, pills, powders, and granules.

[0161] Pharmaceutically acceptable carriers, fillers, and vehicles that may be used in the pharmaceutical compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphoric acid, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0162] Pharmaceutical compositions containing one or more conjugates can be designed for oral or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, inhalation / aerosolized, intraarterial, intrathecal, intratumoral, intracapsular, peritumoral, intraperitomeal, intracavity, intrapleural) administration. When administered orally, the pharmaceutical composition can be in the form of a pill, tablet, or capsule. Compositions suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, or solutes that render the formulation isotonic with the blood of the intended recipient. The formulations can be provided in single-dose or multi-dose containers, for example in sealed ampoules and vials, and can be stored (e.g., in a lyophilized condition) requiring only the addition of a sterile liquid carrier (e.g., water for injection or saline) immediately prior to use. In some embodiments, the formulations can be provided in a form requiring only the addition of a sterile carrier (e.g., water or saline) and the desired radionuclide(s). Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets.

[0163] In some examples, a pharma- ceutical acceptable composition comprising one or more conjugates described herein can be administered locally or systemically. For example, the compositions provided herein can be administered systemically by intravenous injection or transfusion. For example, the compositions provided herein can be administered locally (e.g., intratumorally, intramuscularly, intradermally, or subcutaneously). For example, intra-arterial injection can be used to locally direct the composition (e.g., injection into the hepatic artery to target cancer in the liver). In some examples, the compositions provided herein can be administered systemically, orally, or by injection to a mammal (e.g., a human patient).

[0164] An effective amount of a composition comprising one or more conjugates can be any amount that provides an anti-tumor response (e.g., slowing, stopping, or reversing tumor growth by halting tumor cell proliferation and / or killing tumor cells) without causing significant toxicity to the patient. For example, an effective amount of a conjugate comprising a positron emitting (PET) isotope can be 1 mCi to 20 mCi (e.g., about 1 mCi to about 15 mCi, about 1 mCi to about 10 mCi, about 2 mCi to about 18 mCi, about 3 mCi to about 17 mCi, about 4 mCi to about 18 mCi, about 4 mCi to about 15 mCi, about 5 mCi to about 20 mCi, about 5 mCi to about 15 mCi, about 10 mCi to about 20 mCi, about 15 mCi to about 20 mCi). In some embodiments, an effective amount of a conjugate comprising a beta-emitting isotope may be, for example, about 10 mCi to 1.5 Ci (1,500 mCi) per cycle (e.g., about 15 mCi to about 1,400 mCi, about 25 mCi to about 1,500 mCi, about 50 mCi to about 1,250 mCi, about 75 mCi to about 1,500 mCi, about 100 mCi to about 1,000 mCi, about 100 mCi to about 1,400 mCi, about 150 mCi to about 1,250 mCi, about 200 mCi to about 1,200 mCi, about 300 mCi to about 1,100 mCi, about 400 mCi to about 1,000 mCi, about 500 mCi to about 1,500 mCi, about The amount of the active ingredient may be from about 600 mCi to about 1,400 mCi, from about 700 mCi to about 1,300 mCi, from about 800 mCi to about 1,200 mCi, or from about 1,000 mCi to about 1,500 mCi.In some embodiments, an effective amount of a conjugate comprising a gamma-emitting isotope (e.g., a SPECT agent) can be, for example, about 0.1 mCi to about 40 mCi (e.g., about 0.2 mCi to about 40 mCi, about 0.5 mCi to about 35 mCi, about 0.5 mCi to about 25 mCi, about 1 mCi to about 35 mCi, about 1 mCi to about 30 mCi, about 2 mCi to about 38 mCi, about 3 mCi to about 30 mCi, about 4 mCi to about 35 mCi, about 4 mCi to about 35 mCi, about 5 mCi to about 50 mCi, about 6 mCi to about 60 mCi, about 7 mCi to about 70 mCi, about 8 mCi to about 80 mCi, about 9 mCi to about 90 mCi, about 10 mCi to about 100 mCi, about 12 mCi to about 120 mCi, about 14 mCi to about 140 mCi, about 16 mCi to about 160 mCi, about 18 mCi to about 180 mCi, about 19 mCi to about 200 mCi, about 21 mCi to about 210 mCi, about 22 mCi to about 220 mCi, about 23 mCi to about 230 mCi, about 24 mCi to about 240 mCi, about 25 mCi to about 25 mCi, about 26 mCi to about 25 mCi, about 27 mCi to about 25 mCi, about 28 mCi to about 25 mCi, about

[0113] The amount of isocitrate may be from about 1 mCi to about 35 mCi, from about 5 mCi to about 40 mCi, from about 5 mCi to about 35 mCi, from about 5 mCi to about 30 mCi, from about 5 mCi to about 25 mCi, from about 5 mCi to about 20 mCi, from about 10 mCi to about 30 mCi, from about 15 mCi to about 40 mCi, from about 20 mCi to about 40 mCi, or from about 25 mCi to about 40 mCi. In some embodiments, an effective amount of a conjugate comprising an alpha-emitting isotope is, for example, about 0.05 mCi to 100 mCi per cycle (e.g., about 0.05 to about 90 mCi, about 0.1 mCi to about 100 mCi, about 0.2 mCi to about 90 mCi, about 0.5 mCi to about 95 mCi, about 0.5 mCi to about 85 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 85 mCi, about 2 mCi to about 95 mCi, about 3 mCi to about 100 mCi, about 1 ... The amount of the active ingredient may be from about 5 mCi to about 90 mCi, from about 4 mCi to about 85 mCi, from about 4 mCi to about 80 mCi, from about 5 mCi to about 100 mCi, from about 5 mCi to about 85 mCi, from about 5 mCi to about 70 mCi, from about 5 mCi to about 60 mCi, from about 5 mCi to about 50 mCi, from about 10 mCi to about 100 mCi, from about 15 mCi to about 60 mCi, from about 20 mCi to about 80 mCi, or from about 25 mCi to about 100 mCi).

[0165] In some embodiments in which more than one conjugate is administered, the effective amount of each conjugate may differ, for example, if it is desired to use an alpha-emitting isotope for therapy and a positron-emitting isotope for imaging, different amounts of the conjugates may be administered.

[0166] For example, an effective amount of one or more conjugates described herein can be administered per dose (e.g., daily, weekly, monthly, bimonthly, or quarterly doses) to an average-sized human (e.g., a human of about 75-85 kg). In some cases, a single dose of the conjugate can be followed by a 2-16 week (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 week) washout period to monitor the patient for adverse effects (e.g., by monitoring complete blood count, white blood cell count, platelet count, hemoglobin level, or bone marrow damage) before repeating the dose. Each administration and washout period is referred to as a treatment cycle.

[0167] If a particular mammal fails to respond to a particular amount of therapeutic agent conjugate, or the calculated amount of drug reaching the target tumor is too low, the amount of conjugate injected in the next cycle can be increased, for example by 2-fold. After receiving this relatively high amount, the mammal can be monitored for both responsiveness to treatment and toxic symptoms, and adjustments can be made accordingly. The effective amount can remain constant, or can be adjusted as a sliding scale or variable dose, depending on the mammal's response to treatment. Various factors can affect the actual effective amount used for a particular application. For example, frequency of administration, duration of treatment, use of multiple therapeutic agents, route of administration, and severity of condition may require an increase or decrease in the actual effective amount administered.

[0168] The frequency of administration of the conjugates described herein can be any frequency that provides an anti-tumor response (e.g., halting tumor growth or killing tumor cells) without causing significant toxicity to the mammal. For example, the frequency of administration of the conjugates can be about once a day, once a month, once every six weeks, once every two months, or about once every three months, or about once every 16 weeks. The frequency of administration of the conjugates described herein can remain constant during the treatment period or can be variable (e.g., more frequent administration with less toxicity). As described above, a course of treatment with a composition comprising a conjugate can include a drug holiday. For example, a single administration of a composition comprising one or more conjugates can be followed by a drug holiday of 2 to 6 weeks (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 weeks), and such a regimen can be repeated multiple times. As with the effective amount, various factors may influence the actual frequency of administration used for a particular application, for example, the effective amount, duration of treatment, use of multiple therapeutic agents, route of administration, and severity of the condition may require increased or decreased frequency of administration.

[0169] The effective period for administering a composition comprising one or more conjugates can be any period that provides an anti-tumor response (e.g., halting tumor growth or killing tumor cells) in a mammal identified as having cancer without causing significant toxicity to the mammal. In some cases, the effective period can vary from a few days to a few months. In general, the effective period for providing an anti-tumor response (e.g., halting tumor growth or killing tumor cells) in a mammal identified as having cancer can be within a period of about 6 weeks to about 10 months. Several factors can affect the actual effective period used for a particular treatment. For example, the effective period can vary depending on the frequency of administration, the effective amount, the use of multiple therapeutic agents, the route of administration, and the severity of the condition being treated.

[0170] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims. EXAMPLES

[0171] Example 1 – Diamsar and TCMC platforms for polypeptide conjugation As shown in Figure 3, the diamsar (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexazabicyclo[6.6.6]-eicosane-1,8-diamine (SarAr) with CN=6)-TCMC (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10 tetraazacyclododecane with coordination number (CN) 8, NO) platform can be easily conjugated to any peptide or antibody at room temperature (heating up to 37 °C may be required). TCMC can be conjugated with Pb and Diamsar can be conjugated with Cu, and both complexation reactions can be carried out at room temperature to 37 °C. If necessary, the chain length of the third arm containing the NCS group can be adjusted / extended to alleviate any potential steric hindrance. Competitive conjugation of Pb and Cu can be tested in the presence of both chelators at any given pH, buffer and temperature to confirm conjugation. Due to the lipophilic nature of Diamsar, the overall lipophilic and related properties of the peptide are expected to increase with the addition of Diamsar-TCMC conjugation.

[0172] Example 2 – NOTA and TCMC platforms for polypeptide conjugation As shown in Figure 4, the NOTA (p-SCN-Bn-NOTA, chemically 1,4,7-triazacyclononane-1,4,7-triacetic acid, NO3O3 with CN=6, CN=6)-TCMC platform can be easily conjugated to any peptide or antibody at room temperature (heating up to 37°C may be required). TCMC can be conjugated with Pb and NOTA can be conjugated with Cu, and both complexation reactions can be carried out at room temperature to 37°C. If necessary, the chain length of the third arm containing the NCS group can be adjusted / extended to alleviate any potential steric hindrance. Competitive conjugation of Pb and Cu can be tested at any given pH, buffer and temperature in the presence of both chelators to confirm the conjugation.

[0173] Example 3 – Diamsar and TCMC platforms for dual polypeptide conjugation As shown in Figure 5, the diamsar-TCMC platform can be easily conjugated to any peptide or antibody at room temperature (heating up to 37°C may be required). TCMC can be conjugated to Pb and Diamsar can be conjugated to Cu, with both complexation reactions being possible at room temperature to 37°C. If necessary, the chain length of the third arm containing the NCS group can be adjusted / extended to alleviate any potential steric hindrance. Competitive conjugation of Pb and Cu can be tested at any given pH, buffer and temperature in the presence of both chelators to confirm conjugation. Due to the lipophilic nature of Diamsar, the overall lipophilic and related properties of the peptide are expected to increase with the addition of Diamsar-TCMC conjugation. This approach allows for dual conjugation of peptides and antibodies, facilitating enhanced binding to target receptors.

[0174] Example 4 – NOTA and TCMC platforms for dual polypeptide conjugation As shown in Figure 6, the NOTA-TCMC platform can be easily conjugated to any peptide or antibody at room temperature (heating up to 37°C may be necessary). TCMC can be conjugated to Pb and NOTA can be conjugated to Cu, and both complexation reactions can be performed at room temperature to 37°C. If necessary, the chain length of the third arm containing the NCS group can be adjusted / extended to alleviate any potential steric hindrance. Competitive conjugation of Pb and Cu can be tested at any given pH, buffer and temperature in the presence of both chelators to confirm conjugation. This approach allows dual conjugation of peptides and antibodies, facilitating enhanced binding to target receptors.

[0175] Example 5 – Conjugate 1 64 Cu label As shown in FIG. 10, conjugate 1 and 64 A high performance liquid chromatography (HPLC) method was developed for Cu conjugate 1 (Figure 14). The method used a Schmadzu HPLC system equipped with dual UV and radioactivity detectors. The method was developed and optimized using a Phenomenex reversed phase HPLC column (C-18) (Luna 5 μm C18(2) 100Å LC column 250 x 4.6mm) (00G-4252-e0) with a UV wavelength of 254 nm. For the sample analysis, a 20 μL injection loop was attached and used for all the analyses at room temperature. For the mobile phase, a dual solvent system was used consisting of solvent A as 0.1% trifluoroacetic acid (TFA) in acetonitrile and solvent B as 0.1% TFA in water. For peak separation, a gradient method as described in Tables 12A and 12B was used with a mobile phase flow rate of 1.1 mL / min. Table 11 shows the results of the HPLC method. Figure 11 shows the HPLC calibration curves for the various concentrations shown in Table 12A. For analysis, the compounds were dissolved in water to generate a calibration curve and estimate the specific radioactivity of the synthesized compounds. Different retention times were observed depending on the chemical nature of the compounds.

[0176] [Table 11]

[0177] [Table 12A]

[0178] [Table 12B]

[0179] In addition, unlabeled 64 A HPLC method for Cu was developed. 64 The HPLC trace of Cu is shown. The method used a Schmadzu HPLC system equipped with dual UV and radioactivity detectors. The method was developed and optimized using a Phenomenex reversed phase HPLC column (C-18) (Luna 5 μm C18(2) 100Å LC column 250x4.6mm) (00G-4252-e0) with a UV wavelength of 254 nm. For analyte analysis, a 20 μL injection loop was attached and used for all analyses at room temperature. For the mobile phase, a dual solvent system was used consisting of solvent A as 0.1% trifluoroacetic acid (TFA) in acetonitrile and solvent B as 0.1% TFA in water. For peak separation, a gradient method was used as described in Table 14B below with a mobile phase flow rate of 1.0 mL / min. Table 13 shows the unlabeled 64 The results of the HPLC method for Cu are shown. Unlabeled Cu was measured using silica gel as the solid phase and 0.1 M sodium citrate as the mobile phase. 64 A thin layer chromatography (TLC) method for Cu was developed. 64 The TLC trace for Cu is shown and Table 14A shows the results.

[0180] [Table 13]

[0181] [Table 14A]

[0182] [Table 14B]

[0183] As shown in FIG. 14, conjugate 1 64 Label with Cu, 64 Cu-conjugate 1 was formed. It was generated from a cyclotron and formulated in 0.1 M hydrochloric acid. 64 Conjugate-1 was radiolabeled with Cu-64 using [Cu]CuCl2. Different amounts (50 μg, 100 μg) of conjugate-1 were used to 64 After the addition of Cu]CuCl2, the pH was adjusted to 5.0 with 0.1 M sodium acetate. The resulting reaction mixture was stirred at room temperature for different periods such as 10 min, 20 min, 30 min, and 40 min to optimize the radiolabeling yield with reaction time. The progress and yield of the reaction were monitored by iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) and radioactive thin layer chromatography (r-TLC) using 0.1 M sodium citrate as the mobile phase. Under this r-TLC condition, unconjugated (free) 64 Cu migrates to the solvent front of r-TLC and is radiolabeled 64 Cu-conjugate-1 remains at the origin of the r-TLC plate. Based on our tested radiolabeling conditions, the above reaction achieved >99% radiolabeling yield in 10 minutes and at all other time points by stirring at room temperature at pH 5.0 using sodium acetate as the reaction buffer. Our radiolabeling yield as a function of reaction time, temperature, and mass of starting conjugate-1 is summarized in Table 16. Radiolabeled 1 was determined by radio-HPLC. 64 The formation of Cu-conjugate-1 was also confirmed.

[0184] It is a product 64 The TLC trace of Cu-conjugate 1 is shown in Figure 15. TLC was performed using a silica gel solid phase and a 0.1 M sodium citrate mobile phase. Table 15 shows the TLC results.

[0185] [Table 15]

[0186] unlabeled 64 The same HPLC method used for Cu was used. 64 It was also used for Cu-conjugate 1. 64 FIG. 1 shows an HPLC trace of Cu-conjugate 1.

[0187] Various reaction conditions were used 64 The radiolabeling yields for Cu-conjugate 1 are shown in Table 16 below. 64 Molar radioactivity of Cu-conjugate 1 (A m ) was 0.325 GBq / μmol.

[0188] [Table 16]

[0189] Example 6 – Conjugate 2 64 Cu-labeled As shown in FIG. 18, conjugate 2 64 Label with Cu, 64 Cu-conjugate 2 was formed. It was generated from a cyclotron and formulated in 0.1 M hydrochloric acid. 64 Conjugate 2 was radiolabeled with Cu-64 using [Cu]CuCl2. Different amounts (50 μg, 100 μg) of conjugate 2 were used to 64The pH was adjusted to 5.0 using 0.1 M sodium acetate after the addition of Cu]CuCl2. The resulting reaction mixture was stirred at room temperature for different time points such as 10, 20 and 30 minutes to optimize the radiolabeling yield as a function of reaction time. The progress and yield of the reaction was monitored by iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) and radioactive thin layer chromatography (r-TLC) using 0.1 M sodium citrate as the mobile phase. Based on our tested radiolabeling conditions, the above reaction achieved >99% radiolabeling yield in 10 minutes and at all other time points by stirring at room temperature at pH 5.0 using sodium acetate as the reaction buffer. Our radiolabeling yield as a function of reaction time, temperature and mass of starting conjugate 2 is summarized in Table 22. Radiolabeled 100% 100% 15 ... 64 The formation of Cu-conjugate 2 was also confirmed.

[0190] As shown in Figure 17, an HPLC method for conjugate 2 was developed. The method used a Schmadzu HPLC system equipped with dual UV and radioactivity detectors. The method was developed and optimized using a Phenomenex reversed phase HPLC column (C-18) (Luna 5μm C18(2) 100Å LC column 250 x 4.6mm) (00G-4252-e0) with a UV wavelength of 254 nm. For sample analysis, a 20μL injection loop was attached and used for all analyses at room temperature. For the mobile phase, a dual solvent system was used consisting of solvent A as 0.1% trifluoroacetic acid (TFA) in acetonitrile and solvent B as 0.1% TFA in water. For peak separation, we used the gradient method described in Table 18B with a mobile phase flow rate of 1.0 mL / min. Table 17 below shows the results of the HPLC method. Conjugate 2 was tested at various concentrations (Table 18A below). Figure 19 shows the HPLC calibration curves of various concentrations of conjugate 2. For analysis, the compounds were dissolved in water to generate a calibration curve and estimate the specific radioactivity of the synthesized compounds. Different retention times were observed depending on the chemical nature of the compounds.

[0191] [Table 17]

[0192] [Table 18A]

[0193] [Table 18B]

[0194] Using a silica gel solid phase and a 0.1M sodium citrate mobile phase, 64 The TLC trace of Cu-conjugate 2 is shown in Figure 20. Table 19 shows the TLC results.

[0195] [Table 19]

[0196] The same HPLC method used for conjugate 2 was used 64 It was also used for Cu-conjugate 2. 64 The r-HPLC traces of the Cu-peptide conjugates are shown in Tables 20 and 21. 64 HPLC results for Cu-conjugate 2 are shown.

[0197] [Table 20]

[0198] [Table 21]

[0199] 64 Cu-conjugate 2 was tested for radiolabeling yield using a variety of reaction conditions (Table 22 below). 64 Molar radioactivity of Cu-conjugate 2 (A m ) was in the range of 0.8 to 1.35 GBq / μmol.

[0200] [Table 22]

[0201] 64 The stability of Cu-Conjugate 2 was tested at various time points using the same HPLC method as Conjugate 2. The time points included: 40 min (Tables 23-24 and Figure 22), 2 hours (Tables 25-26 and Figure 23), 4 hours (Tables 27-28 and Figure 24), and 8 hours (Tables 29-30 and Figure 25).

[0202] [Table 23]

[0203] [Table 24]

[0204] [Table 25]

[0205] [Table 26]

[0206] [Table 27]

[0207] [Table 28]

[0208] [Table 29]

[0209] [Table 30]

[0210] 64 By TLC using the same TLC method as for Cu-conjugate 1, 64 The stability of Cu-conjugate 2 was also analyzed at various time points, including: 40 min (Table 31 and Figure 26), 2 hours (Table 32 and Figure 27), 4 hours (Table 33 and Figure 28), and 8 hours (Table 34 and Figure 29).

[0211] [Table 31]

[0212] [Table 32]

[0213] [Table 33]

[0214] [Table 34]

[0215] 64 The stability of Cu-conjugate 2 was tested in mouse and human serum at 37° C. using the Rad-iTLC method. Approximately 1.0 mL of mouse serum was extracted from mouse blood and approximately 1.0 mL of human serum was extracted from blood obtained from the Mayo Clinic blood bank and radiolabeled. 64 The stability of Cu-conjugate 2 was measured. The obtained mouse and human serum was separately distributed into 100 μL aliquots in 1.5 mL microcentrifuge tubes (n=3). 64 Cu-conjugate 2 was added to each 100 μL serum aliquot and mixed thoroughly. From this mixture, a small amount of the reaction mixture was removed using a glass capillary tube and immediately spotted onto an iTLC plate as the T=0 time point for analysis (n=3). The remaining portion of the reaction mixture was incubated at 37° C. for up to 2 hours, and small aliquots were removed after 1 and 2 hours of incubation and analyzed using radioactive thin layer chromatography (r-TLC). 64 The stability of Cu-conjugate 2 was analyzed over time. To perform the r-TLC analysis, iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) was used as the solid phase and 0.1 M sodium citrate was used as the mobile phase. Under these r-TLC conditions, unconjugated (free) 64 Cu migrates to the solvent front of r-TLC and is radiolabeled 64Cu-conjugate 2 remains at the origin of the r-TLC plate. Based on the relative % of radioactivity at the origin and solvent front, 64 Cu-conjugate 2 and free 64 The % Cu was determined as shown in Tables 35 and 36. The results of the stability study are shown in Table 35 below. 64 Cu-conjugate 2 was found to be stable in mouse serum for up to 2 h.

[0216] [Table 35]

[0217] [Table 36]

[0218] 64 The cellular uptake of Cu-conjugate 2 was examined using LNCaP cells. 64 The cellular uptake of Cu-conjugate 2 was tested using LNCaP cells. LNCaP cells were obtained from American Type Culture Collection (Manassas, VA) and cultured in Corning® BioCoat™ poly-lysine 6-well plates (Corning, Glendale, AZ) in a CO2 incubator at 37°C in complete Roswell Park Memorial Institute (RPMI) 1640 medium with 10% fetal bovine serum (FBS) (Gibco-ThermoFisher Scientific, Waltham, MA) and 1x penicillin / streptomycin (Gibco-ThermoFisher Scientific, Waltham, MA). On the day of the uptake experiment, the cell culture medium in the wells culturing the cells was changed to pre-incubation medium (RPMI 1640 with 5% bovine serum albumin (BSA)) and the cells were pre-incubated for 60 min. After pre-incubation, the cells were incubated at 37°C for 1 h at 4°C for 1 h at 4°C for 1 h at 4°C. 64The cells were re-incubated with Cu-conjugate 2 (1.4±0.22 MBq / well at the beginning of incubation) in RPMI1640 medium with 5% BSA at 37° C. for 60 min. 64 After incubation with Cu-conjugate 2, cells were washed three times with chilled phosphate buffered saline (PBS) with or without 10 μM 2-(phosphonomethyl)pentane-1,5-dioic acid (PMPA), a potent PSMA inhibitor. Cells washed with 10 μM PMPA showed 64 Internalization of Cu-conjugate 2 provided information about the contributed uptake, whereas cells washed without PMPA 64 This gave an estimate of the uptake contributed by both internalization and cell membrane binding of Cu-conjugate 2. For negative controls, cells were exposed to 100 μM PMPA during the pre-incubation and incubation steps. After a final wash, cells were harvested from the wells and radioactivity was counted in a gamma counter. Uptake was calculated according to the following formula: % uptake = (decay-corrected radioactivity in cells after washing / decay-corrected radioactivity in incubation medium) x 100. Molar radioactivity 64 Cu-conjugate 2 was 1.35 GBq / μmol. The concentration per well was 1.52 nmol, with 6.5 x 10 per well in a 6-well plate. 5 The % cell uptake is shown in FIG.

[0219] Normal nude mice (strain: 002019, NU / J) 2 hours after injection 64 The in vivo evaluation of Cu-conjugate 2 is shown in Figures 31 and 32. 64 MicroPET imaging was performed on normal mice with Cu-conjugate 2 at different time intervals and is shown in FIG. 64 MicroPET imaging was performed at different time intervals on normal mice (strain: 002019, NU / J) with Cu-conjugate 2 and athymic nude mice bearing LNCaP tumors and is shown in FIG. 33 and FIG. 56. 64Cu-conjugate 2 (5.64±0.25 MBq, 52 GBq / μmol; n=3) was injected into normal mice and athymic nude mice bearing LNCaP tumors. PET images (10 min static) were acquired 30, 60, and 120 min after injection using a small animal PET system (Sofie BioSystems Genesys4, Culver City, CA, USA). Standard uptake values ​​(SUV), maximum standard uptake values ​​(SUV max ) and mean standard uptake value (SUV mean The acquired PET images were analyzed using the image analysis software AMIDE (Amide's Medical Imaging Data Examiner) for calculation of uptake as SUV. After the final image acquisition, the animals were euthanized and the tumor tissue and major organs of interest, such as kidneys, were removed for gamma counting for ex-vivo biodistribution. Uptake in the tissues of interest as SUV was calculated according to Loening AM and Gambhir SS. AMIDE: a free software tool for multimodality medical image analysis. Mol Imaging 2003; 2:131-7, as follows: SUV of tissue of interest = ((activity in tissue of interest / mL) / (injection dose)) x animal weight. 64 Cu-conjugate 2 was found to accumulate in the proximal tubules (Figure 34) in the kidney, an area known to have high prostate-specific membrane antigen (PSMA) expression. 64 It was shown that Cu-conjugate 2 was well tolerated by the animals and reached the expected organs of the body.

[0220] Example 7 – Conjugate 1 203 Pb-labeled The above 64 Using the same HPLC method as used for Cu, unlabeled 203 / 212 The Pb was analyzed and the HPLC trace is shown in Figure 35 and Table 37 below.

[0221] [Table 37]

[0222] unlabeled 203 / 212 A TLC method for Pb was developed that included a silica gel (iTLC) stationary phase and a 0.15 M NH4Ac, pH 4.0 mobile phase. The TLC results are shown in Figure 36 and Table 38.

[0223] [Table 38]

[0224] As shown in FIG. 37, conjugate-1 203 Label with Pb 203 Pb-conjugate-1 was formed. Conjugate 1 was 203 To radiolabel with Pb, 203 Radioactive Pb-203 as [Pb]PbCl2 was used as an alternative radioisotope to Pb-212 to test the feasibility of radiolabeling. Approximately 100 μg of conjugate-1 was used, and [ 203 After the addition of [Pb]PbCl2, the pH was adjusted to 6.0 using 0.15 M ammonium acetate (pH 6.5-7.0), and the resulting reaction mixture was stirred at 37 °C for 10 and 30 min to optimize the radiolabeling yield with reaction time. The progress and yield of the reaction were monitored by radioactive thin layer chromatography (r-TLC) using iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) as the solid phase and 0.15 M NH4Ac, pH 4.0 as the mobile phase. Under these r-TLC conditions, unconjugated (free) 203 Pb migrates to the solvent front of r-TLC and is radiolabeled 203Pb-conjugate 1 remains at the origin of the r-TLC plate. Based on the radiolabeling conditions tested, the above reaction achieved >99% radiolabeling yield in 30 min using 0.15 M ammonium acetate as the reaction buffer at pH 6.0 and stirring at 37° C. The radiolabeling yield as a function of reaction time, temperature, and mass of starting conjugate-1 is summarized in Table 39. Radiolabeled product was determined by radio-HPLC. 203 The formation of Pb-conjugate 1 was also confirmed. 64 The same HPLC method used for Cu was used. 203 It was also used for Pb-conjugate 1. Figure 38 shows 203 HPLC trace of Pb-conjugate 1 is shown. 203 Molar radioactivity of Pb-conjugate 1 (A m ) was 0.107 GBq / μmol.

[0225] [Table 39]

[0226] Example 8 – Conjugate 2 203 Pb-labeled As shown in Figure 39, conjugate 2 203 Label with Pb 203 Pb-conjugate 2 was formed. 203 To radiolabel with Pb, radioactive Pb-203 was added to [ 203 The feasibility of radiolabeling was tested using PbCl2, an alternative radioisotope to Pb-212. Approximately 200 μg of conjugate 2 was used, and [ 203After the addition of [Pb]PbCl2, the pH was adjusted to 6.0 using 0.15 M ammonium acetate (pH 6.5-7.0). The resulting reaction mixture was stirred at 37 °C for 30 min. The progress and yield of the reaction were monitored by radioactive thin layer chromatography (r-TLC) using iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) as the solid phase and 0.15 M NH4Ac, pH 4.0 as the mobile phase. Under these r-TLC conditions, unconjugated (free) 203 Pb migrates to the solvent front of r-TLC and is radiolabeled 203 Pb-conjugate 2 remains at the origin of the r-TLC plate. Based on the radiolabeling conditions we tested, the above reaction achieved >99% radiolabeling yield in 30 minutes by stirring at 37° C. at pH 6.0 using 0.15 M ammonium acetate as the reaction buffer. The radiolabeling yield as a function of reaction time, temperature, and mass of starting conjugate 2 is summarized in Table 43. Radiolabeled product was determined by radio-HPLC. 203 The formation of Pb-conjugate 2 was also confirmed.

[0227] 203 Using the same TLC method used to analyze Pb-conjugate-1, 203 Pb-conjugate 2 was analyzed and the TLC trace is shown in Figure 40 and Table 40 below.

[0228] [Table 40]

[0229] 203 Using the same HPLC method used to analyze Pb-conjugate 1, 203 Pb-conjugate 2 was analyzed and the HPLC trace is shown in Figure 41 and Tables 31 and 32 below.

[0230] [Table 41]

[0231] [Table 42]

[0232] unlabeled 203 / 212 Using the TLC method developed for Pb 203 The reaction yields of Pb-conjugate 2 were determined and are shown in Table 43 below. 203 Molar radioactivity of Pb-conjugate 2 (A m ) was 0.299 GBq / μmol.

[0233] [Table 43]

[0234] unlabeled 203 By TLC using the same TLC method as for Pb, 203 The stability of Pb-conjugate 2 was also analyzed at various time points. These time points included: 40 min (Figure 42), 2 hours (Figure 43), 4 hours (Figure 44), and 21 hours (Figure 45). The results showed that: 203 It was shown that Pb-conjugate 2 was stable for up to 21 hours.

[0235] Example 9 - 64 Cu and 203 Mixed labeling of conjugate 2 with Pb As shown in Figure 46, conjugate 2 203 Pb and 64 Cu-labeled and mixed-labeled conjugates, 64 Cu / 203 Pb-conjugate 2 was formed. Conjugate 2 is designed as a theranostic molecule that serves as both an imaging molecule and a radiotherapy molecule. 203 Pb radioisotope and 64 However, in this experiment, 212As a substitute isotope for Pb 203 First, conjugate 2 was used. 203 Radioactively labeled with Pb, so that radioactive Pb-203 is [ 203 Approximately 200 μg of conjugate 2 was used, and [ 203 After the addition of [Pb]PbCl2, the pH was adjusted to 6.0 using 0.15 M ammonium acetate (pH 6.5-7.0). The resulting reaction mixture was stirred at 37 °C for 20-30 min. The progress and yield of the reaction was monitored by radioactive thin layer chromatography (r-TLC) using iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) as the solid phase and 0.15 M NH4Ac, pH 4.0 as the mobile phase. Under these r-TLC conditions, unconjugated (free) 203 Pb migrates to the solvent front of r-TLC and is radiolabeled 203 Pb-conjugate 2 remains at the origin of the r-TLC plate. Based on the radiolabeling conditions tested, the above reaction achieved >99% radiolabeling yield in 20-30 minutes with 0.15 M ammonium acetate as the reaction buffer, pH 6.0, and stirring at 37° C. The TLC results are shown in FIG. 47. 203 After confirming the radioactive labeling with Pb, the reaction temperature was adjusted to room temperature and the [ 64 Cu-64 as Cu]CuCl2 was added. The pH was adjusted to 5.0 using 0.1 M sodium acetate, and the resulting reaction mixture was stirred for an additional 10 min at room temperature. The progress and yield of the reaction was monitored by iTLC (silica gel coated on paper, Agilent Technologies Inc., Santa Clara, CA) and radioactive thin layer chromatography (r-TLC) using 0.1 M sodium citrate as the mobile phase. Under these r-TLC conditions, unconjugated (free) 64 Cu migrates to the solvent front of r-TLC and is radiolabeled 64Cu-conjugate 1 remains at the origin of the r-TLC plate. Based on the radiolabeling conditions tested, the above reaction yielded >99% yield in 10 min. 64 Radiolabeling of Cu was achieved and the TLC results are shown in Figure 48.

[0236] 64 Cu / 203 Pb-conjugate 2 was stability tested using TLC to measure stability. TLC analysis was performed using two different solvent systems. The first solvent system was 0.1 M sodium citrate and the second solvent system was 0.15 M NH4Ac, pH 4.0. Stability was measured at various time points, e.g.: 1 hour (Figure 49), 4 hours (Figure 50), and 21 hours (Figure 51). 64 Cu / 203 Pb-conjugate 2 was found to be stable for up to 21 hours at room temperature.

[0237] Example 10 64 Mixed labeling of conjugate 2 with Cu and non-radioactive Pb As shown in Figure 46, conjugate 2 was synthesized by the reaction of 64 Cu-labeled and mixed-labeled conjugates, 64 Cu / Pb-conjugate 2 was formed. 64 Cu / Pb-conjugate 2 was formed in two steps. First, PbCl2 was added to 0.15 M ammonium acetate buffer (pH 6.5-7) and the mixture was stirred at 37 °C for about 20 min at pH 6 to form complexed Pb-conjugate 2. Second, Pb-conjugate 2 was added to 0.1 M sodium acetate buffer (pH 5.0), followed by 64 CuCl2 was added to the mixture. The mixture was stirred at room temperature for about 20 minutes at pH 5.

[0238] 64 The Cu / Pb-conjugate 2 was analyzed by TLC using a silica gel solid phase and a 0.1 M sodium citrate mobile phase. The TLC results are shown in Figure 53 and Table 44 below.

[0239] [Table 44]

[0240] 64 Cu / Pb-conjugate 2, unlabeled 64 The compounds were analyzed by HPLC using the same HPLC method as used in the Cu HPLC method. The HPLC trace is shown in Figure 54. Molar radioactivity (A m ) was 52GBq / μM.

[0241] 64 The in vitro uptake of Cu / Pb-conjugate 2 was tested. The cell line used was LNCaP in Matrigel, the incubation temperature was 37°C, the incubation time was 1 hour, and the incubation medium was RPMI1640 + 5% bovine serum albumin. 64 Compared to Cu-conjugate 2, 64 The in vitro uptake results of Cu / Pb-conjugate 2 were normalized to 64 The increased cellular uptake of Cu / Pb-conjugate 2 was demonstrated (Figure 55).

[0242] 64 The in vivo uptake of Cu / Pb-conjugate 2 was examined. PET images were taken in a LNCaP tumor model. 64 The results showing the in vivo uptake of Cu / Pb-conjugate 2 are shown in Figures 56 and 57 and Table 45.

[0243] [Table 45]

[0244] Molar radioactivity (A m The effect of 1,2-dichlorophenyl 1,3-dichlorophenyl 1,4 ...64 A comparison of the uptake of Cu / Pb-conjugate 2 (n=3 per group) was performed. The results showed that the uptake in tumor-bearing mice was significantly higher than that in normal mice (Figure 57). The molar specific radioactivity was measured as radioactivity / micromoles of ligand. 64 The tumor-specific SUVmax and SUVmean of the in vivo uptake of Cu / Pb-conjugate 2 were investigated. In the LNCaP tumor model, the A of 52 GBq / μmol was m have 64 In vivo uptake was performed using Cu / Pb-conjugate 2. The results are shown in Figures 60, 61 and 62 and Table 45 below, and Figures 63, 64 and 65 and Table 46 below. The results shown in Figures 60-62 and Table 46 were obtained from experiments using different animals with different tumor locations than the animals used in the experiment that produced the results shown in Figures 63-65 and Table 47. The results showed the intensity of the imaging probe, highlighting the heterogeneity of the tumor.

[0245] [Table 46]

[0246] [Table 47]

[0247] Example 11 64 In vitro uptake of Cu-conjugate 2 64 The in vitro uptake of Cu-conjugate 2 was tested. The cell line used was LNCaP in Matrigel, the incubation temperature was 37 °C, A m The concentration was 0.254 GBq / μmol, the concentration / well was 2.33 nmol, and the number of cells per well was 1.97 x 10 6 The incubation time was 1 hour, and the incubation medium was RPMI1640 + 5% bovine serum albumin. 64The results of in vitro uptake of the Cu / Conjugate 2 conjugate are shown in FIG.

[0248] In normal and tumor-bearing mice 64 A comparison of ex-vivo biodistribution uptake of Cu-conjugate 2 was performed, and the results are shown in Figure 57.

[0249] In the LNCaP tumor model 64 Organ specific uptake of Cu-conjugate 2 was assessed. The results are shown in Figure 67. SUV ratios of organ / tissue vs. muscle uptake were determined. The results are shown in Figure 68.

[0250] In addition, the mouse 64 MicroPET images of tumor-bearing mice were taken after injection of Cu-conjugate 2. The microPET images of the mice are shown in FIG.

[0251] Example 12 –Synthesis of alpha-PET conjugates for enhanced tumor uptake, retention and replication As shown in Figure 70, two PSMA targeting vectors (e.g., lysine and glutamic acid covalently linked together by a urea bond) or analogs thereof are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a phthalate-based aromatic moiety with three functional groups (two for tethering the PSMA vector and the third for binding a dual chelator for imaging and radiotherapy applications). The dual targeting conjugate contains a six-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0252] As shown in Figure 71, two PSMA targeting vectors are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a diethylenetriamine-based aliphatic moiety with three functional groups, two for tethering the PSMA vector and the third for binding a dual chelator for imaging and radiotherapy applications. The dual targeting conjugate contains a six-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0253] As shown in Figure 72, two PSMA targeting vectors are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a phthalate-based aromatic moiety with three functional groups, two for tethering the PSMA vector and a third for attaching a dual chelator for imaging and radiotherapy applications. The dual targeting conjugate is 223 Ra, 225 Ac, and 213 It contains a MACROPA chelator that allows for the chelation of additional alpha-emitting radioisotopes such as Bi. The dual targeting vector contains a 6-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0254] As shown in Figure 73, two PSMA targeting vectors are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a diethylenetriamine-based aliphatic moiety with three functional groups, two for tethering the PSMA vector and the third for attaching a dual chelator for imaging and radiotherapy applications. The dual targeting conjugate is 223 Ra, 225 Ac, and 213It contains a MACROPA chelator that allows for the chelation of an additional alpha-emitting radioisotope such as Bi. The dual targeting conjugate contains a 6-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0255] As shown in Figure 74, two PSMA targeting vectors are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a phthalate-based aromatic moiety with three functional groups, two for tethering the PSMA vector and a third for attaching a dual chelator for imaging and radiotherapy applications. The dual targeting vector is 223 Ra, 225 Ac, and 213 The dual targeting conjugates include a MACROPA chelator that allows for the chelation of an additional alpha-emitting radioisotope, such as Bi. 89 It further contains a DFO chelator for conjugation with longer-lived PET isotopes such as Zr. The dual targeting conjugate contains a 6-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0256] As shown in Figure 75, two PSMA targeting vectors are mixed together to form a dual targeting conjugate. The dual targeting conjugate is synthesized using a diethylenetriamine-based aliphatic moiety with three functional groups, two for tethering the PSMA vector and the third for attaching a dual chelator for imaging and radiotherapy applications. The dual targeting conjugate is 223 Ra, 225 Ac, and 213 The dual targeting conjugates include a MACROPA chelator that allows for the chelation of an additional alpha-emitting radioisotope, such as Bi. 89It further contains a DFO chelator for conjugation with longer-lived PET isotopes such as Zr. The dual targeting conjugate contains a 6-carbon alkyl chain as a spacer between the chelator and the PSMA-binding vector to avoid steric hindrance in target binding and synthesis.

[0257] As shown in Figure 76, a single PSMA targeting vector is mixed with a chelator to form a single targeted conjugate. A single targeted conjugate is 223 Ra, 225 Ac, and 213 The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeted conjugate with a MACROPA chelator is believed to enhance the uptake and retention of the designed compound in the tumor and / or may not require longer exposure for effective radiotherapy.

[0258] As shown in Figure 77, a single PSMA targeting vector is mixed with a chelator to form a single targeted conjugate. A single targeted conjugate is 223 Ra, 225 Ac, and 213 The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeting conjugate having a MACROPA chelator and an additional chelator is believed to enhance tumor uptake and retention of the designed compound and / or may not require longer exposure for effective radiotherapy.

[0259] As shown in Figure 78, a single FAPI targeting vector is mixed with a chelator to form a single targeted conjugate. A single targeted conjugate is 223 Ra, 225 Ac, and 213The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeted conjugate with a MACROPA chelator is believed to enhance the uptake and retention of the designed compound in the tumor and / or may not require longer exposure for effective radiotherapy.

[0260] As shown in Figure 79, a single FAPI targeting vector is mixed with a chelator to form a single targeted conjugate. A single targeted conjugate is 223 Ra, 225 Ac, and 213 The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeting conjugate with a MACROPA chelator and an additional DFO chelator is believed to enhance the uptake and retention of the designed compound in tumors and / or may not require longer exposure for effective radiotherapy.

[0261] As shown in Figure 80, a single octreotide targeting vector is mixed with a chelator to form a single targeted conjugate. The single targeted conjugate is 223 Ra, 225 Ac, and 213 The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeted conjugate with a MACROPA chelator is believed to enhance the uptake and retention of the designed compound in the tumor and / or may not require longer exposure for effective radiotherapy.

[0262] As shown in Figure 81, a single octreotide targeting vector is mixed with a chelator to form a single targeted conjugate. The single targeted conjugate is 223 Ra, 225 Ac, and 213The MACROPA chelator allows for the chelation of additional alpha-emitting radioisotopes such as Bi. A single targeting conjugate with a MACROPA chelator and an additional DFO chelator is believed to enhance the uptake and retention of the designed compound in tumors and / or may not require longer exposure for effective radiotherapy.

[0263] Example 13 64 Dual labeling of FAP-targeted multifunctional chelators with both Cu and non-radioactive Pb As shown in Figure 82, conjugate 3 64 Label with Cu, 64 Cu-FAPI conjugate ( 64 A stock solution of conjugate 3 (FAPI-NOTA-TCMC) with a concentration of 1.0 mg / mL was prepared by using 300 μg of conjugate 3 in 300 μL of 0.1 M NaOAc (pH 5.0) prior to radiolabeling. 64 Cu]CuCl2 was reconstituted in 2.0 mL of 0.1 M NaOAc (pH 5.0) (Figure 82). As shown in Figure 83, conjugate 3 was 64 Labeled with Cu and non-radioactive Pb 64 Cu / Pb-FAPI conjugate ( 64 For double labeling with Cu-64 and non-radioactive Pb, the radiolabeling reaction was carried out with 10 μg of conjugate 3 (FAPI-NOTA-TCMC) dissolved in 0.1 M NaOAc (pH 5.0), 10 μL of 0.15 M NH4Ac (pH 7.0), and 1.8 μL of PbCl2 (1.0 mg / mL in 0.15 M NH4Ac (pH 7.0)) were added, the reaction mixture was stirred at 37 °C for 20 min, and then 200 μL of [ 64Cu]CuCl2 was added. The resulting reaction mixture with a final reaction pH of about 5.0 (4.7-5.0) was stirred for another 10 min at room temperature (Figure 83). The progress of the reaction and the reaction yield were measured using Rad-TLC. For rad-TLC, i-TLC (silica gel coated paper TLC) was used, with 0.1 M sodium citrate (pH 4.5) as the mobile phase. Similarly, conjugate 3 (FAPI-NOTA-TCMC) was separately radiolabeled with Cu-64 using 10 μg FAPI and 200 μL of Cu-64 for 10 min at room temperature, with a final reaction pH of 4.4-4.7 and a radiolabeling yield of nearly 100%.

[0264] Radiolabeling reactions have also been successfully performed by reversing the order of labeling, meaning labeling with Pb followed by Cu-64 (or vice versa) using appropriate temperature and pH. 64 The compound was successfully characterized by rad-TLC, HPLC and rad-HPLC using the reference compound of Cu]CuCl2 and control TLC. As shown in Figure 84, after complex formation, 64 The UV HPLC trace of Cu-conjugate 3 was achieved using a solvent gradient of 0.0 (95% B) -12:00 (45% B) - 23 (95% B) -30 (stop) (0.1% TFA water, solvent B). Figure 85 shows the free [ 64 CuCl2] and Figure 86 shows the rad-TLC trace. 64 FIG. 87 shows the rad-TLC trace of Cu-conjugate 3. 64 The rad-TLC of Cu / Pb-conjugate 3 was shown. Double labeling after complexation as shown in Figures 88 and 89. 64 Both US and radioactive HPLC traces were taken to analyze the purity of Cu / Pb-conjugate 3.

[0265] A single peak was observed by HPLC, indicating the formation of a complete complex (Figure 84). 64 Cu]CuCl2 and 64Comparison of the rad-TLC traces of Cu-conjugate 3 revealed a significant shift in the radiation population (Figures 85 and 86). 64 A single peak was observed by rad-TLC for Cu-conjugate 3, indicating complete complexation.

[0266] Analysis by HPLC with both UV and radioactive detection identified one major peak that constituted approximately 94% of the product (Figures 88 and 89), indicating highly efficient complex formation. 64 Cu]CuCl2 and 64 Further confirmation was provided by comparison of the rad-TLC traces for Cu / Pb-conjugate 3 (Figure 87). Analysis of the complex revealed a single pure peak.

[0267] Example 14 64 Dual labeling of somatostatin-targeted multifunctional chelators with both Cu and non-radioactive Pb As shown in Figure 90, conjugate 4 64 Labeled with Cu and non-radioactive Pb 64 Cu / Pb-FAPI conjugate ( 64 Prior to radiolabeling, a stock solution of conjugate 4 (octreotide-NOTA-TCMC) with a concentration of 1.0 mg / mL was prepared by using 300 μg of conjugate 4 in 300 μL of 0.1 M NaOAc (pH 5.0) or water. 64 Cu]CuCl2 was reconstituted in 2.0 mL of 0.1 M NaOAc, pH 5.0. For double labeling with Cu-64 and non-radioactive Pb, radiolabeling reactions were performed with 10 μg, 20 μg, and 50 μg of conjugate 4 dissolved in 0.1 M NaOAc, pH 5.0 or in water, and 10 μL of 0.15 M NH4Ac, pH 7.0, and 1.8 μL PbCl2 (1.0 mg / mL in 0.15 M NH4Ac, pH 7.0) were added. The reaction mixture was stirred for 20 min at 37 °C, after which 25 μL or 50 μL of [64 Cu]CuCl2 (Figure 90) was added. The resulting reaction mixture with a final reaction pH of about 5.0 (4.7-5.0) was stirred for another 10 min at room temperature. The progress of the reaction and the reaction yield were measured using Rad-TLC. For rad-TLC, i-TLC (silica gel coated paper TLC) was used, with 0.1 M sodium citrate (pH 4.5) as the mobile phase. Similarly, conjugate 4 (octreotide-NOTA-TCMC) was separately radiolabeled with Cu-64 using 10 μg of conjugate 4 (octreotide-NOTA-TCMC) and 50 μL of Cu-64 at room temperature for 10 min, with a final reaction pH of 4.4-4.7, with a radiolabeling yield of nearly 100%. Radiolabeling reactions have also been successfully carried out by reversing the order of labeling, meaning labeling with Pb followed by Cu-64 (or vice versa) using appropriate reaction temperatures and pH. 64 It was successfully characterized by rad-TLC, HPLC and rad-HPLC using the reference compound of [Cu]CuCl2 and control TLC. As shown in Figures 91 and 92, 64 The UV and radioactive HPLC traces of Cu / Pb-conjugate 4 were analyzed. 64 Cu]CuCl2 (Figure 93) and 64 Rad-TLC traces were taken of both Cu / Pb-conjugates 4 (Figure 94).

[0268] After the reaction shown in FIG. 64 The Cu / Pb-conjugate 4 complex was verified by both HPLC and rad-TLC. By HPLC, analysis with both UV and radioactive detection identified one major peak that constituted about 94% of the product (Figures 91 and 92), indicating highly efficient complex formation. The reaction efficiency was [ 64 Cu]CuCl2 and 64 Further confirmation was obtained by comparison of the rad-TLC traces for Cu / Pb-conjugate 4 (Figures 93 and 94). Analysis of the complex reveals a single, pure peak.

[0269] Example 15 212 Pb-conjugate 2 synthesis and radionuclide therapy of prostate tumors method Tumor model creation: Prostate cancer cell line, LNCaP, was obtained from American Type Culture Collection (Manassas, VA). LNCaP tumor models were created using male athymic nude mice obtained from Charles Rivers Laboratories (Wilmington, MA) or The Jackson Laboratory (Bar Harbor, ME) according to well-established LNCaP subcutaneous tumor protocols (Horoszewicz et al. Prog Clin Biol Res. 1980; 37:115-32; Horoszewicz et al. Cancer Res. 1983 Apr;43(4):1809-18). On the day of cell implantation, LNCaP cells in culture medium were trypsinized and washed twice in serum-free RPMI-1640 medium. Cells were then cultured at 5×10 6 The cells were resuspended in serum-free RPMI-1640 medium at a concentration of 100 μL cells / 100 μL. 100 μL LNCaP cell suspension was injected subcutaneously between the shoulder blades of each animal. Physical examination of the animals and 64 The presence of subcutaneous tumors was confirmed by PET imaging using a Cu-conjugated 2 PSMA imaging probe. For confirmation based on PET imaging, approximately 100 μCi 64 Cu-conjugate 2 was injected intravenously via tail vein injection, and 15 min of static PET images were acquired 1 h postinjection using a small animal Micro-PET / X-ray system (Sofie BioSystems Genesys4, Culver City, CA, USA). PET images were visualized and analyzed using MIM 7 software (MIM Software Inc., Cleveland, OH, USA).

[0270] 212 Pb-conjugate 2 radionuclide therapy: physical examination, and 64The presence of PSMA+ LNCaP tumors in the animals was confirmed after confirmation by PET imaging with Cu-conjugate 2. On the day of radionuclide treatment, 4.2 mCi [ 212 Pb]PbCl2 was received from the vendor in 2.1 mL sodium acetate (1 M, pH 6.0) solution. 212 To prepare Pb-conjugate 2, add 1.0 mL of [ 212 The reaction mixture was prepared by aliquoting [Pb]PbCl2 (2.1 mCi) into a 5.0 mL V-vial followed by the addition of 25 μg of conjugate 2. The reaction mixture was then stirred for 20 min at 37 °C. 100% chelation efficiency was confirmed using rad-TLC with ammonium acetate (0.15 M, pH 4.0) as the mobile phase.

[0271] After 100% chelation, 4.0 mL of deionized water was added. 212 Pb-conjugate 2 was added to the reaction mixture to give a formulation of 2.1 mCi / 5.0 mL or 50 μCi / 100 μL at pH 6.0. After dilution, the rad-TLC was analyzed again for 100% chelation efficiency. 212 A single bolus dose of [Pb]Pb-NSN-24901 (0.096±0.002 mCi, n=4 mice) was intravenously injected via tail vein injection into each athymic nude mouse bearing a PSMA+LNCaP tumor. The animals were then observed and tumor size was assessed by: 212 The tumor size was measured 3, 5, 9, 14, and 18 days after injection of Pb-conjugate 2. The total reduction in tumor size or percentage of tumor regression was calculated as follows: 212 Compared to tumor size observed before Pb-conjugate 2 treatment, 212 Tumor size (cm) 3, 5, 9, 14, and 18 days after Pb-conjugate 2 injection 2 ) after 18 days of treatment, along with a physical exam showing no tumors. 64 The absence of tumor was also confirmed using Cu-conjugated 2 PSMA imaging.

[0272] result 212 Formation and stability of Pb-conjugate 2:212 Pb-conjugate 2 was prepared according to the scheme shown in Figure 95. The formation of the complex was confirmed by rad-TLC shown in Figure 97, and the comparative free [ 212 Pb]PbCl2 is shown in Figure 96. Free [ 212 Pb]PbCl2, [ 212 Comparison to rad-TLC for [Pb]Pb-NSN-24901 indicates 100% complex formation. The stability of the complex over time was also monitored by rad-TLC. The complex remained 95.0% intact after 2 hours of incubation (Figure 98) and was 89.7% intact after 22 hours of incubation (Figure 99). This suggested that the complex was stable enough for the time required to be used therapeutically.

[0273] Alpha Emission 212 Radionuclide treatment of prostate tumors with Pb-conjugate 2: After establishing LNCaP tumors in nude mice, these were 212 Pb-conjugate 2 (0.096±0.002 mCi, n=4 mice) and tumor size was monitored over time by both physical examinations (Figure 100). The tumor in one mouse was treated with PET imaging radionuclide-loaded, 100 μg / mL of the same molecule used therapeutically. 64 The tumors were also monitored by PET imaging using Cu-conjugate 2 (Figure 101). Both physical examination and PET imaging indicate that radionuclide therapy successfully reduced tumor size over time. The results for individual mice are summarized in Table 48.

[0274] [Table 48]

[0275] Example 16 – Exemplary Conjugates This example provides structures of exemplary conjugates described herein.

[0276] Exemplary Conjugates for Targeting Prostate Cancer

[0277] Conjugate Aa

[0278] [ka]

[0279] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0280] Conjugate A2

[0281] [ka]

[0282] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0283] Conjugate A3

[0284] [ka]

[0285] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0286] Conjugate A4

[0287] [ka]

[0288] In some examples, this structure may have various combinations of Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes) along with Zr-89 (radioactive and non-radioactive).

[0289] Conjugate A5

[0290] [ka]

[0291] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0292] Conjugate A6

[0293] [ka]

[0294] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0295] Conjugate A7

[0296] [ka]

[0297] In some examples, this structure may have various combinations of Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes) along with Zr-89 (radioactive and non-radioactive).

[0298] Conjugate A8

[0299] [ka]

[0300] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0301] Conjugate A9

[0302] [ka]

[0303] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0304] Conjugate A10

[0305] [ka]

[0306] In some examples, this structure may have various combinations of Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes) along with Zr-89 (radioactive and non-radioactive).

[0307] Exemplary Conjugates for Targeting Neuroendocrine Tumors

[0308] Conjugate B1

[0309] [ka]

[0310] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0311] Conjugate B2

[0312] [ka]

[0313] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0314] Conjugate B3

[0315] [ka]

[0316] In some examples, this structure may have various combinations of Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes) along with Zr-89 (radioactive and non-radioactive).

[0317] Exemplary Conjugates for Targeting Fibroblast Activation Protein (FAP)

[0318] Conjugate C1

[0319] [ka]

[0320] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0321] Conjugate C2

[0322] [ka]

[0323] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0324] Conjugate C3

[0325] [ka]

[0326] In some examples, this structure may have various combinations of Zr-89 (radioactive and non-radioactive) along with Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0327] Exemplary Conjugates for Targeting Folate

[0328] Conjugate D1

[0329] [ka]

[0330] In some examples, the structure may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive.

[0331] Conjugate D2

[0332] [ka]

[0333] In some examples, this structure may have various combinations with Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0334] Conjugate D3

[0335] [ka]

[0336] In some examples, this structure may have various combinations of Zr-89 (radioactive and non-radioactive) along with Ac-225 / Ac-226 / Ra-223 (both radioactive and non-radioactive isotopes).

[0337] Example 17 – Exemplary Binding Moieties This example provides amino acid sequences of exemplary binding moieties that may be used in the conjugates described herein.

[0338] [Table 49] TIFF2024523344000097.tif227160TIFF2024523344000098.tif102159

[0339] [Table 50] TIFF2024523344000100.tif232159TIFF2024523344000101.tif232160TIFF2024523344000102.tif26159

[0340] [Table 51] TIFF2024523344000104.tif228159TIFF2024523344000105.tif142159

[0341]

Table 52

[0342]

Table 53

[0343]

Table 54

[0344]

Table 55

[0345]

Table 56

[0346]

Table 57

[0347]

Table 58

[0348] Other embodiments Although the present invention has been described with reference to its detailed description, it is understood that the foregoing description is intended to be illustrative, and not limiting, of the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A conjugate comprising two or more chelators and a linking moiety, wherein one of said chelators is a chelator for an imaging isotope and one of said chelators is a chelator for a radiotherapy isotope, and said chelator and said linking moiety are of the moiety of formula (I): 【Chemical 1】 (wherein: Each X is N, P, P(=O), CR N , and the moiety of formula (i): [Chemical Formula 2] is independently selected from; x 1 , x 2 , x 3 , and x 4 each of which independently represents a point of attachment of the moiety of formula (I) to the chelator or the linking moiety; L 1 、L 2 、L 3 、and L 4 each of which is independently selected from C(=O), C(=S), N(R N ), O, S, S(=O), S(=O) 2 , -CR N =NR N (-C 1-3 alkylene - O -) x (-O - C 1-3 alkylene -) x , -C 1-3 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, C 3-10 cycloalkylene, C 6-10 arylene, 5 - to 14 - membered heteroarylene, and 4 - to 10 - membered heterocycloalkylene, where each x is independently an integer from 1 to 10, and each of said -C 1-3 alkylene -, C 2-6 alkenylene, C 2-6 alkynylene, C 3-10 cycloalkylene, C 6-10 arylene, 5 - to 14 - membered heteroarylene, and 4 - to 10 - membered heterocycloalkylene is optionally substituted with 1, 2, or 3 substituents independently selected from OH, NO 2 , CN, halo, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, amino, C 1-3 alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 alkoxycarbonyl; y 1 、y 2 、y 3 、and y 4 Each of them is, independently, an integer from 1 to 10; Each R N is independently selected from H, C 1-3 alkyl, and C 1-3 haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5) The conjugate, which is linked via.

2. The conjugate according to claim 1, wherein said isotope used for radiotherapy is an α emitter.

3. the radiation therapy isotope is 225 Ac, 212 Pb, 211 At, 213 Bi, 212 Bi, 211 Bi, 152 / 160 / 161 Tb, 227 Th, 223 Ra, 211 Po, 221 Fr, 217 At, 213 Po, 212 Po, 215 Po, or 177 Lu, the conjugate according to claim 1.

4. The imaging isotope is 68 Ga, 44 Sc, 60 / 61 / 62 / 64 Cu, 84 / 86 / 87 / 89 Zr, 63 Zn, 43 / 44 Sc, 192 / 193 / 194 / 196 Au, 52m Mn, 90 / 92m1 Nb, 51 / 52 Mn, 148 / 151 / 151m / 152 Tb, 45 Ti, 65 / 66 / 67 Ga, 94m Tc, 55 Co, 80 / 81 / 83 Sr, 38 K, 70 / 71 / 72 / 74 As, 81 / 82m Rb, 52 Fe, or 86 Y, and the conjugate according to claim 1.

5. The imaging isotope is 64 Cu, and the radiotherapy isotope is 212 Pb, the conjugate according to claim 1.

6. The conjugate according to claim 1, wherein said imaging isotope is complexed to said chelator of said imaging isotope.

7. The conjugate according to claim 1, wherein said radiotherapy isotope is complexed to said chelator of said radiotherapy isotope.

8. Each of said chelators independently comprises a compound selected from the group consisting of 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), dodecane tetraacetic acid (DOTA), 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC), 1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), and diethylenetraminepentacetic acid (DTPA). The conjugate according to claim 1.

9. The conjugate according to claim 1, wherein said linking moiety is a polypeptide.

10. The conjugate according to claim 9, wherein said polypeptide binds to prostate-specific membrane antigen, somatostatin receptor, or melanocortin-1 receptor.

11. The conjugate according to claim 9, wherein said polypeptide is an antibody.

12. The conjugate according to claim 1, wherein said linking moiety is a small molecule.

13. The conjugate according to claim 12, wherein said small molecule is a glutamate carboxypeptidase II inhibitor.

14. The conjugate according to claim 1, wherein the chelator is covalently bonded to the binding moiety.

15. The conjugate according to claim 1, wherein the chelator and the binding moiety are covalently bonded via a linker.

16. The conjugate according to claim 15, wherein the compound of formula (I) has the following formula: 【Chemical Formula 3】

17. The conjugate according to claim 16, wherein the moiety of formula (I) has any one of the following formulas: 【Chemical Formula 4】 【Chem.】 【Chem.】

18. The conjugate according to claim 1, wherein the chelator and the binding moiety are a moiety of formula (II): 【Chemical Formula 5】 (wherein x 1 represents the binding site of formula (II) to the chelator; x 2 represents the binding point of formula (II) to the chelator or the binding moiety; Each L is independently selected from C(=O), C(=S), N(R N ), O, S, S(=O), S(=O) 2 , -CR N =NR N -, (-C 1-3 alkylene - O -) x , (-O - C 1-3 alkylene -), x , -C 1-3 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, C 3-10 cycloalkylene, C 6-10 arylene, 5 - to 14 - membered heteroarylene, and 4 - to 10 - membered heterocycloalkylene, where each x is independently an integer from 1 to 10, and each of said -C 1-3 alkylene -, C 2-6 alkenylene, C 2-6 alkynylene, C 3-10 cycloalkylene, C 6-10 arylene, 5 - to 14 - membered heteroarylene, and 4 - to 10 - membered heterocycloalkylene is optionally substituted with 1, 2, or 3 substituents independently selected from OH, NO 2 , CN, halo, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, amino, C 1-3 alkylamino, di(C 1-3 alkyl)amino, carboxy, and C 1-3 alkoxycarbonyl; y is an integer from 1 to 30; Each R N is independently selected from H, C 1-3 alkyl, and C 1-3 haloalkyl).

19. The conjugate according to claim 18, wherein the moiety of formula (II) has any one of the following formulas: 【Chemical Formula 6】

20. A composition for use in a method of treating cancer in a mammal in need thereof, wherein the composition comprises the conjugate according to claim 1, wherein the conjugate comprises the imaging isotope complexed to the chelator of the imaging isotope, and wherein the conjugate comprises the radiotherapy isotope complexed to the chelator of the radiotherapy isotope.

21. A composition for use in a method of treating cancer in a mammal, wherein the composition comprises a first conjugate comprising two or more chelators and a binding moiety, wherein one of the chelators is a chelator of an imaging isotope and one of the chelators is a chelator of a radiotherapy isotope, the first conjugate comprises the imaging isotope complexed to the chelator of the imaging isotope, and the method comprises the following: a) administering the composition to the mammal; b) determining the biodistribution of the first conjugate in the mammal; and c) administering to the mammal a second conjugate that is identical to the first conjugate except that it comprises an amount of the radiotherapy isotope complexed to the chelator of the radiotherapy isotope

22. The composition according to claim 21, wherein the method further comprises determining, in the mammal, the in vivo distribution of the second conjugate comprising the imaging isotope complexed to the chelator of the imaging isotope and the radiotherapy isotope complexed to the chelator of the radiotherapy isotope.

23. The composition according to claim 20 or 21, wherein the cancer is selected from the group consisting of prostate cancer, neuroendocrine cancer, colon cancer, lung cancer, pancreatic cancer, melanoma, and lymphoma.

24. The composition according to claim 21 or 22, wherein the second conjugate is the conjugate according to any one of claims 1 to 19.

25. A composition for use in a method of treating cancer in a mammal in need of treatment for cancer, wherein the composition comprises two or more conjugates, where each conjugate comprises two or more chelators and a linking moiety, one of the chelators being a chelator for an imaging isotope and one of the chelators being a chelator for a radiotherapy isotope, one of the conjugates comprising an imaging isotope complexed to the chelator of the imaging isotope, one of the conjugates comprising a radiotherapy isotope complexed to the chelator of the radiotherapy isotype, said composition.

26. The conjugate according to any one of claims 1 to 7, wherein each of the chelators independently comprises a compound selected from the group consisting of NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown-6 (MACROPA).

27. The conjugate according to claim 9, wherein the polypeptide binds to prostate-specific membrane antigen, somatostatin receptor, fibroblast activation protein, or melanocortin-1 receptor.

28. The conjugate according to any one of claims 1 to 8, wherein the conjugate comprises two or more linking moieties.

29. The conjugate according to claim 28, wherein each of the linking moieties is a polypeptide.

30. The conjugate according to claim 29, wherein each of the polypeptides independently binds to prostate-specific membrane antigen, somatostatin receptor, fibroblast activation protein, or melanocortin-1 receptor.

31. The conjugate according to any one of claims 1 to 19, wherein the conjugate comprises three or more chelators.

32. The conjugate according to claim 31, wherein each of the chelators independently comprises a compound selected from the group consisting of NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, NTA, BisTris, EGTA, EDTA, BAPTA, DO2A, DTPA, DO3A, and MACROPA.

33. The conjugate according to claim 1, having the following structure: 【Chemical Formula 7】

34. The conjugate according to claim 1, having the following structure: 【Chemical 8】

35. The conjugate according to claim 1, having the following structure: 【Chemical Formula 9】

36. The conjugate according to claim 1, having the following structure: 【Chemical Formula 10】

37. The conjugate according to claim 1, having the following structure: 【Chemical 11】

38. The conjugate according to claim 1, having the following structure: 【Chemical 12】

39. The conjugate according to claim 1, having the following structure: 【Chemical 13】

40. The conjugate according to claim 1, having the following structure: 【Chemical Formula 14】

41. The conjugate according to claim 1, having the following structure: 【Chemical Formula 15】

42. The conjugate according to claim 1, having the following structure: 【Chemical 16】

43. The conjugate according to claim 1, having the following structure: 【Chemical 17】

44. The conjugate according to claim 1, having the following structure: 【Chemical 18】

45. The conjugate according to claim 1, having the following structure: 【Chemical Formula 19】

46. The conjugate according to claim 1, having the following structure: 【Chemical 20】

47. The conjugate according to claim 1, having the following structure: 【Chemical 21】

48. The conjugate according to claim 1, having the following structure: 【Chemical 22】 The conjugate according to claim 1, having

49. The conjugate has the following structure: 【Chemical 23】 The conjugate according to claim 1, having

50. The conjugate has the following structure: 【Chemical Formula 24】 The conjugate according to claim 1, having

51. The conjugate has the following structure: 【Chemical 25】 The conjugate according to claim 1, having

Images