Methods and materials for combining biologics with multiple chelators

Abstract

Provided herein are conjugates comprising two or more chelators (e.g., a radiotherapeutic isotope chelator and an imaging isotope chelator) containing functional groups capable of stabilizing a radiometal, covalently attached to one or more binding moieties. The conjugates can be used to treat cancerous or non-cancerous conditions, and can function as both imaging and radiotherapeutic molecules when the imaging isotope is complexed to the imaging isotope chelator and the radiotherapeutic isotope is complexed to the radiotherapeutic isotope chelator.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Provisional Application No. 63 / 434,451, filed December 21, 2022. The disclosure of this prior application is considered part of the disclosure of this application and is incorporated herein in its entirety.

[0002] background 1. Technical Field This document relates to conjugates of two or more chelators (e.g., a conjugate of a chelator for an imaging isotope and a chelator for 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 for an isotope used for imaging and one of the chelators is a chelator for an isotope used for radiotherapy. Conjugates in which an imaging isotope and a radiotherapeutic isotope are complexed to a chelator can be administered to a mammal in need of treatment and can serve 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 method for understanding disease pathology, disease progression, and response to radionuclide therapy. It also helps enhance drug discovery and personalize care for patients (e.g., cancer patients) through a 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 unsuitable for imaging due to the unavailability or low doses of suitable positron or photon energy emissions (511 KeV for PET and 100-200 KeV for SPECT). High linear energy transfer (LET) alpha emissions and gamma photons, characteristic x-rays, or bremsstrahlung radiation associated with the decay of the parent alpha-emitting radionuclide are less suitable for quantification of target uptake, dosimetry, and treatment response compared to beta-emitters. Furthermore, even when treatments are performed with imageable beta-emitters, 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 superior. As a result, most research and development, FDA submissions, and clinical programs must rely on estimated biodistribution / dosimetry based on low-quality images or by using surrogate imaging probes (imageable modified drugs). These surrogate imaging probes differ significantly from alpha-emitter therapeutics in various ways, making them less suitable for predicting the biodistribution / dosimetry of alpha-emitting therapeutics. Therefore, 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 for combining (e.g., covalently linking) 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 when a suitable isotope is complexed with the chelator, the resulting conjugate or mixture of conjugates can simultaneously serve as both an imaging molecule and a radiotherapeutic molecule. 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 for an isotope used for imaging (referred to herein as a "chelator for an imaging isotope") and one of the chelators is a chelator for an isotope used for radiotherapy (referred to herein as a "chelator for a radiotherapeutic isotope"). As described herein, the conjugates can be selectively used for imaging or radionuclide therapy, as needed, by selecting either the imaging radionuclide or the therapeutic radionuclide and loading the other chelator with a non-radioactive version of the imaging or therapeutic metal ion, thereby maintaining the same chemical properties of the molecule. By using the same chemical entity, the same biodistribution is maintained, avoiding the use of alternative imaging probes that may have different structures and 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 may be suboptimal for one or both tasks. The present disclosure is based, at least in part, on the recognition that the stability of the radiometal within the chelator moiety can be enhanced by the functional group(s) located on the chelator moiety. As described more fully herein, suitable examples of stability-enhancing functional groups include, for example, amine, hydroxy, amide, carboxy, and other anionic and cationic functional groups capable of forming ligands for radiometals.

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

[0006] To ensure that the biodistribution of the treatment over time that it is radioactive can be imaged and thus dosimetry can be accurately calculated, the conjugates described herein can be designed to match the half-life of the imaging isotope (e.g., a positron emission tomography (PET) isotope or a single photon emission computed tomography (SPECT) isotope) with the physical half-life of the radiotherapeutic isotope (e.g., an alpha- or beta-emitting radionuclide). For example, to ensure that the biodistribution of the treatment over time that it is radioactive can be imaged and thus dosimetry can be accurately calculated, the half-life of the imaging isotope (e.g., a PET isotope or a SPECT isotope), 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. In some embodiments, an optical imaging (near-infrared) probe can be attached to the conjugate. The conjugates and methods described herein provide a robust platform for staging disease, treating disease, monitoring response or progression to treatment, and / or minimizing side effects on healthy organs and tissues, all using versions of the same molecule (chemically and biologically identical). This can be achieved simply by selecting, for the desired use of the conjugate, whether the conjugate described herein is complexed with an imaging isotope and / or a radiotherapeutic isotope, or with non-radioactive versions of these same isotopes (i.e., the radionuclide can be exchanged for a chemically identical non-radioactive isotope with a different nuclear structure). In some embodiments, two or more conjugates 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. Furthermore, the conjugates described herein can contain two or more binding moieties or motifs to enhance uptake in targeted tissues / organs.

[0007] In one general aspect, this document provides a conjugate comprising two or more chelators and a binding moiety (e.g., a targeting vector). In some embodiments, 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 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; At least one chelator in the conjugate is selected from the group consisting of C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH, and C(=O)OR. a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 selected from cycloalkyl, 5-10 membered heteroaryl, and 4-10 membered heterocycloalkyl; provided that the functional groups in said at least one chelator are not all C(=O)NH2 or C(=O)OH; The chelator and the binding moiety are of formula (I):

[0009] [ka] (In the formula: Each X is N, P, P(=O), CR N and the moiety of formula (i):

[0010] [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 the chelator or the 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 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, wherein each x is independently an integer of 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 the group consisting of 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 independently represents an integer from 1 to 10; Each R N is H, C 1-3 Alkyl, and C 1-3independently selected from haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5. wherein the conjugate is linked via

[0011] In some embodiments, the compound of formula (I) has the following formula:

[0012] [ka] It has.

[0013] In some embodiments, the moiety of formula (I) has any one of the following formulas:

[0014] [ka] I have TIFF2026502864000005.tif120147.

[0015] In some embodiments, the chelators of the imaging isotopes are independently selected from the group consisting of 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-hexaazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (diethylenetriaminepentacetic acid), and methylparaben-1,4-diamine (methylparaben-1,4-diamine). acid) (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA), each of which is selected from the group consisting of C(=O)NH2, C(=O)OH, -S(=O)2OH, -S(=O)2NH2, -OP(=O)(OH)2, -P(=O)(OH)2, -OP(=O)(OH)NH2, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH2, and C(=O)OR a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 It is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

[0016] In some embodiments, the chelator of the imaging isotope is substituted with three or four functional groups independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH, provided that at least one functional group is other than C(=O)NH or C(=O)OH.

[0017] In some embodiments, the chelator of the imaging isotope has the following formula:

[0018] [ka] (In the formula, each R 1 , R 2 , and R 3 are independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH;

[0019] [ka] indicates the point of attachment to formula (I) It has.

[0020] In some embodiments, R 1 , R 2 , and R 3 At least one of is other than C(=O)NH2 or C(=O)OH.

[0021] In some embodiments, the imaging isotope is: 133 / 135 La, 133 Ce, 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.

[0022] In some embodiments, the imaging isotope is 64 It is Cu.

[0023] In some embodiments, the chelators of the radiotherapeutic isotopes are independently selected from the group consisting of 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-hexaazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferrocene, and methylparaben. and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA), each of which is selected from the group consisting of C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH, and C(=O)OR. a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 It is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

[0024] In some embodiments, the chelator of the radiotherapeutic isotope is substituted with three or four functional groups independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH, provided that at least one functional group is other than C(=O)NH or C(=O)OH.

[0025] In some embodiments, the chelator of the radiotherapeutic isotope has the following formula:

[0026] [ka] (In the formula, each R 1 , R 2 , R 3 , and R 4 are independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH;

[0027] [ka] indicates the point of attachment to formula (I) It has.

[0028] In some embodiments, R 1 , R 2 , R 3 , and R 4 At least one of is other than C(=O)NH2 or C(=O)OH.

[0029] In some embodiments, the radiotherapeutic isotope is an alpha emitter.

[0030] In some embodiments, the radiotherapeutic isotope is: 223 / 225 / 227 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.

[0031] In some embodiments, the radiotherapeutic isotope is 212 It is Pb.

[0032] In some embodiments, the conjugate has any one of the following formulas:

[0033] [ka] (wherein each R in any of the above formulas 1 , R 2 , R 3 , and R 4 is independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH; R 1 , R 2 , R 3 , and R 4 At least one of is other than C(=O)NH2 or C(=O)OH) is selected from.

[0034] In some embodiments, the binding moiety is selected from a polypeptide, an antibody, a small molecule, an extracellular vesicle, a virus, and a nucleic acid.

[0035] In some embodiments, the polypeptide binds to a prostate-specific membrane antigen, a somatostatin receptor, or a melanocortin-1 receptor.

[0036] In some embodiments, the small molecule is a glutamate carboxypeptidase II inhibitor.

[0037] In some embodiments, the small molecule is glucose or a derivative thereof.

[0038] In some embodiments, the conjugate is any one of the following compounds:

[0039] [ka] TIFF2026502864000012.tif144151TIFF2026502864000013.tif127150TIFF2026502864000014.tif215151 (wherein each R 1 , R 2 , R 3 , and R 4 is independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH; R 1 , R 2 , R 3 , and R 4 At least one of is other than C(=O)NH2 or C(=O)OH) is selected from.

[0040] In some embodiments, 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; the binding moiety is selected from an extracellular vesicle, a virus, a nucleic acid, and glucose or a derivative thereof; The chelator and the binding moiety are of formula (I):

[0041] [ka] (In the formula: Each X is N, P, P(=O), CR N and the moiety of formula (i):

[0042] [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 the chelator or the 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 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, wherein each x is independently an integer of 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 the group consisting of OH, NO, CN, halo, C 1-3Alkyl, 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 independently represents an integer from 1 to 10; Each R N is 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. wherein the conjugate is linked via

[0043] In some embodiments, the binding moiety is an extracellular vesicle.

[0044] In some embodiments, the binding moiety is a virus.

[0045] In some embodiments, the binding moiety is a nucleic acid.

[0046] In some embodiments, the compound of formula (I) has the following formula:

[0047] [ka] It has.

[0048] In some embodiments, the moiety of formula (I) has any one of the following formulas:

[0049] [ka] I have TIFF2026502864000019.tif216144.

[0050] In some embodiments, the chelators of the imaging isotopes are independently selected from the group consisting of 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), 2,2′,2″,2′″-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (DOTA), 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC), 1-N-(4-aminobenzyl)-3,6,1 The compound includes a compound selected from the group consisting of 0,13,16,19-hexazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA).

[0051] In some embodiments, the chelator of the radiotherapeutic isotope independently comprises a compound selected from the group consisting of 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-hexaazabicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA).

[0052] In some embodiments, the chelator for the imaging isotope is 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) and the chelator for the radiotherapeutic isotope is 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC).

[0053] In some embodiments, the imaging isotope is: 133 / 135 La, 133 Ce, 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.

[0054] In some embodiments, the imaging isotope is 64 It is Cu.

[0055] In some embodiments, the radiotherapeutic isotope is an alpha emitter.

[0056] In some embodiments, the radiotherapeutic isotope is: 223 / 225 / 227 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.

[0057] In some embodiments, the radiotherapeutic isotope is 212 It is Pb.

[0058] In some embodiments, the conjugate is any one of the following compounds:

[0059] [ka] Selected from TIFF2026502864000021.tif180142.

[0060] In some aspects of the above embodiments, the binding moiety is selected from an extracellular vesicle, a virus, a nucleic acid, and glucose or a derivative thereof.

[0061] In some embodiments, the present disclosure provides a conjugate described herein, wherein the conjugate comprises an imaging isotope complexed to a chelator for the imaging isotope, and / or the conjugate comprises a radiotherapeutic isotope complexed to a chelator for the radiotherapeutic isotope.

[0062] In some embodiments, the present disclosure provides a method of treating cancer in a mammal in need thereof, the method comprising administering to the mammal a conjugate described herein, wherein the conjugate comprises the imaging isotope complexed to the chelator of the imaging isotope, and the conjugate comprises the radiotherapeutic isotope complexed to the chelator of the radiotherapeutic isotope.

[0063] In some embodiments, the present disclosure provides a method of treating cancer in a mammal, comprising: a) administering to said mammal a first conjugate described herein 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 a second conjugate described herein that is identical to said first conjugate except that it comprises said radiotherapeutic isotope complexed to said chelator of said radiotherapeutic isotope. The method includes:

[0064] 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.

[0065] In some embodiments, the cancer is selected from the group consisting of prostate cancer, neuroendocrine cancer, colon cancer, breast cancer, fibroblast-associated protein cancer, brain cancer, liver cancer, ovarian cancer, thyroid cancer, lung cancer, pancreatic cancer, melanoma, and lymphatic cancer.

[0066] In some embodiments, the cancer is a metastatic tumor.

[0067] In some embodiments, the present disclosure provides a method of treating cancer in a mammal in need thereof, the method comprising administering to the mammal two or more conjugates described herein; wherein each conjugate comprises two or more chelators and a binding moiety, one of said chelators being a chelator for an imaging isotope and one of said 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 radiotherapeutic isotope; The method is provided.

[0068] In some embodiments, the binding moiety is glucose or a derivative thereof.

[0069] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the present invention, 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. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0070] 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 explanation of the drawings]

[0071] [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. [Figure 3A] FIG. 3A includes chemical structures of imaging agents (radiopharmaceuticals containing imageable Cu isotopes) within the scope of the present invention. [Figure 3B]Figure 3B includes chemical structures of radionuclide therapeutic agents (radiopharmaceuticals). The compounds in Figures 3A and 3B represent concepts involving both single and / or dual injections of imaging and radionuclide therapeutic agents, as appropriate. The imaging agent can be separately synthesized with an imaging isotope (e.g., Cu-64) and loaded with a non-radioactive version of a therapeutic isotope (e.g., Pb), or vice versa. After independent synthesis, formulation, and characterization, these agents can be injected into humans as a single injection of either the imaging or therapeutic agent, or a dual / combined injection of both the therapeutic and imaging agent. [Figure 4A] Figure 4A includes the chemical structure of a Pb conjugate with functional groups designed to promote Pb stability. Examples of R1, R2, R3, and R4 include NH2, OH, and NHOH. COR1, COR2, COR3, and COR4 can be any of -SO3H, -SO2NH2, -PO4H2, -PO3HNH2, -CONH2, -COOH, -CONHOH, -CS2H, -COSH, -CSOH, and -CSNH2. Pb can be any isotope of Pb, including but not limited to 212Pb, 203Pb, and Pb, and Cu can be any isotope of Cu, including but not limited to 64Cu, 67Cu, and 61Cu. The targeting vector can be a small molecule, peptide, protein, or antibody, or other biologic, such as, but not limited to, extracellular vesicles, viruses, nucleotides and nucleosides, RNA, or DNA. The targeting vector can target any disease or condition. [Figure 4B]Figure 4B includes the chemical structure of a Cu conjugate with functional groups designed to promote its stability. Examples of R1, R2, and R3 include NH2, OH, and NHOH. COR1, COR2, and COR3 can be any of -SO3H, -SO2NH2, -PO4H2, -PO3HNH2, -CONH2, -COOH, -CONHOH, -CS2H, -COSH, -CSOH, and -CSNH2. Pb can be any isotope of Pb, including but not limited to 212Pb, 203Pb, and Pb, and Cu can be any isotope of Cu, including but not limited to 64Cu, 67Cu, and 61Cu. The targeting vector can be a small molecule, peptide, protein, or antibody, or other biologic, such as, but not limited to, extracellular vesicles, viruses, nucleotides and nucleosides, RNA, or DNA. The targeting vector can target any disease or condition. [Figure 5-1] FIG. 5 includes chemical structures of conjugates with representative examples of binding moieties (eg, targeting vectors such as glucose or its derivatives). [Figure 5-2] This is a continuation of Figure 5-1. [Figure 5-3] This is a continuation of Figure 5-2. [Figure 6A] Figure 6A includes a scheme showing the synthesis of a conjugate for treating small metastatic tumors using alpha-PET technology. The conjugate shown, complexed to a radioisotope, is specifically designed to target small metastatic tumors using a glucose analog. In this design, glucose and glucose analogs act as targeting vectors, and the alpha-PET platform molecule after conjugation facilitates their imaging and radionuclide therapy. [Figure 6B] FIG. 6B includes a scheme showing the conjugation of an alpha-PET platform moiety to an extracellular vesicle and radiolabeling. [Figure 7] FIG. 7 contains a scheme showing the conjugation and radiolabeling of alpha-PET platform moieties to various types of viruses. [Figure 8] FIG. 8 includes a scheme showing the conjugation and radiolabeling of various aptamers with the alpha-PET platform moiety. [Figure 9] FIG. 9 shows the structure of [64Cu]Cu-NOTA-TCMC-adenoassociated virus ([64Cu]Cu-NOTA-TCMC-AAV). [Figure 10] FIG. 10 shows radio-TLC chromatograms showing A. 64Cu and B. [64Cu]Cu-NOTA-TCMC-AAV9 using 100 mM sodium citrate (pH 5.0) as the mobile phase. [Figure 11] Figure 11 shows representative maximum intensity projection (MIP)-PET / X-ray atlas images showing the biodistribution of [Cu]Cu-NOTA-TCMC-AAV9 (approximately 2.0 μCi) in BALB / c mice at various time points after injection. Li = liver, SUV = standardized uptake value. [Figure 12] FIG. 12 shows the chemical structure of [64Cu]Cu-NOTA-TCMC-extracellular vesicles ([64Cu]Cu-NOTA-TCMC-EV). [Figure 13] FIG. 13 shows radio-TLC chromatograms showing A. 64Cu and B. [64Cu]Cu-NOTA-TCMC-EV using 100 mM sodium citrate (pH 5.0) as the mobile phase. [Figure 14] Figure 14 shows representative maximum intensity projection (MIP)-PET / X-ray atlas images showing the biodistribution of [Cu]Cu-NOTA-TCMC-EVs (approximately 23.25 μCi) in BALB / c mice at various time points after injection. Li = liver, Gu = intestine, SUV = standardized uptake value. DETAILED DESCRIPTION OF THE INVENTION

[0072] Detailed Description This document provides conjugates 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 (e.g., N',N'-bis(2-aminoethyl)ethane-1,2-diamine, etc.) that can act as a linker can selectively react with two different chelators, one for the imaging isotope and one for the radiotherapeutic isotope, to generate a dual chelator compound. The dual chelator compound can be modified to make it suitable for reaction 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 conjugates in which two or more chelators are covalently linked to one or more binding moieties or motifs. Only one functional group on the targeting binding moiety (e.g., primary NH) is required to generate the conjugate. As described below, the combination of chelator and isotope can be varied depending on the needs of the treatment or imaging method.

[0073] In some embodiments, the chelator is a moiety of formula (I):

[0074] [ka] (In the formula: Each X is N, P, P(=O), CR N and the moiety of formula (i):

[0075] [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(RN ), 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, wherein each x is independently an integer of 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 the group consisting of 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 independently represents an integer from 1 to 10; Each R N is 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

[0076] In some embodiments, X is N.

[0077] In some embodiments, X is P.

[0078] In some embodiments, X is P(=O).

[0079] In some embodiments, X is CR N is.

[0080] In some embodiments, X is a moiety of formula (i).

[0081] In some embodiments, X is N and CR N is selected from.

[0082] In some embodiments, X is N, CR N and the moiety of formula (i).

[0083] In some embodiments, each L 1 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 In some embodiments, the moiety (L 1 ) y1 is of the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0084] In some embodiments, each L 2 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 In some embodiments, the moiety (L 2 ) y2 is of the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0085] In some embodiments, each L 3 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10In some embodiments, the moiety (L 3 ) y3 is of the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0086] In some embodiments, each L 4 is C(=O), C(=S), NH, O, -C 1-3 Alkylene- and C 6-10 In some embodiments, the moiety (L 4 ) y4 is of the formula NHC(=S)NH or C 6-10 Arylene-C 1-3 Contains at least one alkylene- moiety.

[0087] 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.

[0088] In some embodiments, R N is H. In some embodiments, R N is C 1-3 In some embodiments, R N is H and C 1-3 alkyl.

[0089] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0090] In some embodiments, the compound of formula (I) has the following formula:

[0091] [ka] It has.

[0092] In some embodiments, the compound of formula (I) has the following formula:

[0093] [ka] It has.

[0094] In some embodiments, the compound of formula (I) has the following formula:

[0095] [ka] It has.

[0096] In some embodiments, the moiety of formula (I) has any one of the following formulas:

[0097] [ka] TIFF2026502864000028.tif92138.

[0098] In some embodiments, the moiety of formula (II):

[0099] [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 the 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 , -C1-3 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, wherein each x is independently an integer of 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 the group consisting of 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 is H, C 1-3 Alkyl, and C 1-3 haloalkyl) is used to attach the chelator and / or to attach the chelator to the binding moiety.

[0100] 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.

[0101] 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 of the formula NHC(=S)NH or C 6-10 Arylene-C1-3 Contains at least one alkylene- moiety.

[0102] 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 is H and C 1-3 alkyl.

[0103] In some embodiments, the moiety of formula (II) has any one of the following formulae:

[0104] [ka] It has.

[0105] In some embodiments, the chelator may be attached to the binding moiety via a cleavable linker. As used herein, the term "cleavable linker" 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 specific compound (e.g., a compound present in the body, such as a specific protease) such that the linker is cleaved 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, for example, Zheng et al., Acta Pharm Sin B. 2021 Dec;11(12):3889-3907 and Tsuchikama 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 esterases native to the cellular environment or can be hydrolyzable by a neutral or acidic buffered environment. The phosphate bond can be cleavable by 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 prolonged retention in the body, while the chelator and associated radionuclide can be rapidly excreted.

[0106] In some embodiments, a cleavable linker can connect a binding moiety to one or more chelators. For example, a cleavable linker can connect a binding moiety to two chelators. In some embodiments, cleavage of the linker can separate one or more chelators from the binding moiety.

[0107] 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.

[0108] Various aryl, heteroaryl, cycloalkyl, and heterocycloalkyl rings are described in various places herein. 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 a pyridin-2-yl, pyridin-3-yl, or pyridin-4-yl ring.

[0109] The term "aromatic" refers to a carbocyclic or heterocyclic ring having one or more polyunsaturated rings that have aromatic character (i.e., have (4n+2) delocalized π (pi) electrons, where n is an integer).

[0110] 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.

[0111] As used herein, the phrase "optionally substituted" means unsubstituted or substituted. The substituents are independently selected, and the substitution can 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.

[0112] 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 Examples include:

[0113] 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-chained or branched, having n to m carbon atoms. Examples of alkyl moieties include, but are not limited to, chemical groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, and sec-butyl; higher homologs such as 2-methyl-1-butyl, n-pentyl, 3-pentyl, n-hexyl, and 1,2,2-trimethylpropyl. 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.

[0114] As used herein, "C n-m The term "haloalkyl," used alone or in combination with other terms, refers to an alkyl group having from 1 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 exclusively fluorinated. In some embodiments, the alkyl group has 1 to 6, 1 to 4, or 1 to 3 carbon atoms.

[0115] 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.

[0116] 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.

[0117] As used herein, "C n-m The term "alkylene," used alone or in combination with other terms, refers to a divalent alkyl-linked 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 "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.

[0118] As used herein, "C n-mThe 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.

[0119] 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.

[0120] As used herein, the term "amino" refers to a group of formula -NH2.

[0121] 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.

[0122] 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.

[0123] 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.

[0124] 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.

[0125] 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.

[0126] 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 spirocycles. 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 and cyclohexane. 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 carbon atoms (C 3-10 In some embodiments, cycloalkyl can have C 3-10 In some embodiments, cycloalkyl is a monocyclic or bicyclic cycloalkyl. 3-7 Monocyclic 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.

[0127] 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-membered 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 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 heteroaryl ring is a heteroaryl having a ring with 6 ring atoms, wherein one or more (e.g., 1, 2, or 3) ring atoms are independently selected from N, O, and S. Exemplary 6-membered heteroaryls are pyridyl, pyrazinyl, pyrimidinyl, triazinyl, and pyridazinyl.

[0128] 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, benzazepine, and the like. The ring-forming carbon atoms and heteroatoms of a heterocycloalkyl group can be optionally substituted with one or two independently selected oxo or sulfido groups (e.g., C(O), S(O), C(S), or S(O)). A heterocycloalkyl group can be bonded through a ring-forming carbon atom or ring-forming heteroatom. In some embodiments, a heterocycloalkyl group contains zero to three double bonds. In some embodiments, a heterocycloalkyl group contains zero to two 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, and azepine. Heterocycloalkyl groups containing fused aromatic rings can be bonded through any ring-forming atom, including a ring-forming atom of the fused aromatic ring. In some embodiments, a heterocycloalkyl is a monocyclic 4- to 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 monocyclic 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.

[0129] In certain places, definitions or embodiments refer to specific 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.

[0130] As used herein, the term "compound" is meant to encompass all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. Compounds identified herein by name or structure as one particular tautomeric form are intended to encompass other tautomeric forms unless otherwise specified.

[0131] The compounds described herein can be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of the present invention containing 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, can 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 can 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.

[0132] The compounds provided herein also encompass tautomeric forms. Tautomeric forms result from the swapping 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 in 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 locked into one form by appropriate substitution.

[0133] In some embodiments, each of the chelators may independently be, for example, NOTA, DOTA, TCMC, DiAmSar, HBED, DFO, DTPA, H4pypa, NTA, BisTris, EGTA, EDTA, BAPTA, DO2A, DO3A, and MACROPA. Generally, the combination of chelators for the imaging isotope and the therapeutic isotope may be selected for a particular application. For example, in some embodiments, one chelator may be DiAmSar and one chelator may be TCMC. In some embodiments, one chelator may be NOTA and one chelator may be TCMC. In some embodiments, each of the chelators may independently be a supermagnetic iron oxide nanoparticle (SPION). In some embodiments, the SPION may 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 herein by reference in their entireties.

[0134] In some embodiments, the conjugate may contain three or more chelators. For example, in some embodiments, the conjugate may contain three chelators, 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 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 contain 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.

[0135] In some embodiments, the chelator in the conjugate comprises one or more (1, 2, 3, 4, 5, or 6; or 2, 3, 4, or 5; or 3, 4, or 5; or 3 or 4) functional groups different from a carboxylic acid or amide (or the corresponding hydroxyl or NH2 group). In some embodiments, the chelator is selected from the group consisting of S(=O)2OH, -S(=O)2NH2, -OP(=O)(OH)2, -P(=O)(OH)2, -OP(=O)(OH)NH2, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH2, and C(=O)OR a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10In some embodiments, the chelator comprises at least one functional group selected from -S(=O)2OH, -S(=O)2NH2, -OP(=O)(OH)2, -P(=O)(OH)2, -OP(=O)(OH)NH2, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH2.

[0136] In some embodiments, the chelator is selected from the group consisting of C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH, and C(=O)OR. a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 In some embodiments, the functional groups in the chelator are not all C(=O)NH2. In some embodiments, the functional groups in the chelator are not all C(=O)OH.

[0137] In some embodiments, the chelator in the conjugate comprises at least one S(=O)2OH. In some embodiments, the chelator in the conjugate comprises at least one -S(=O)2NH2. In some embodiments, the chelator in the conjugate comprises at least one -OP(=O)(OH)2. In some embodiments, the chelator in the conjugate comprises at least one -P(=O)(OH)2. In some embodiments, the chelator in the conjugate comprises at least one -OP(=O)(OH)NH2. In some embodiments, the chelator in the conjugate comprises at least one -C(=O)NHOH. In some embodiments, the chelator in the conjugate comprises at least one C(=S)SH. In some embodiments, the chelator in the conjugate comprises at least one C(=O)SH. In some embodiments, the chelator in the conjugate comprises at least one -C(=S)OH. In some embodiments, the chelator in the conjugate comprises at least one -C(=S)NH2. In some embodiments, the chelator in the conjugate is at least one C(=O)OR a where R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 It is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

[0138] In some embodiments, the chelator (e.g., of an imaging isotope or a radiotherapeutic isotope) is independently selected from the group consisting of 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-hexaaza The compound includes a compound selected from the group consisting of bicyclo[6.6.6]eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA).

[0139] In some embodiments, the chelator (e.g., of an imaging isotope or a radiotherapeutic isotope) is independently selected from the group consisting of 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 (HB), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA), each of which is selected from the group consisting of C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH, and C(=O)OR. a wherein R a is C 1-6Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 It is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

[0140] In some embodiments, a chelator (e.g., of an imaging isotope or a radiotherapeutic isotope) is substituted with three or four functional groups independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH, provided that at least one functional group is other than C(=O)NH or C(=O)OH.

[0141] In some embodiments, the chelator (e.g., of an imaging isotope or a radiotherapeutic isotope) has the following formula:

[0142] [ka] (In the formula, each R 1 , R 2 , and R 3 are independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH;

[0143] [ka] indicates the point of attachment to formula (I) It has.

[0144] In some embodiments, R 1 , R 2 , and R3 At least one of is other than C(=O)NH2 or C(=O)OH.

[0145] In some embodiments, the chelator (e.g., of an imaging isotope or a radiotherapeutic isotope) has the following formula:

[0146] [ka] (In the formula, each R 1 , R 2 , R 3 , and R 4 are independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH;

[0147] [ka] indicates the point of attachment to formula (I) It has.

[0148] In some embodiments, R 1 , R 2 , R 3 , and R 4 At least one of is other than C(=O)NH2 or C(=O)OH.

[0149] The imaging isotopes 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., 223 / 225 / 227 Ac, 212 Pb, 211 At, 213 Bi, 212 Bi, 211 Bi, 152 / 160 / 161 Tb, 227 Th, 223 Ra, 211 Po, 221Fr, 217 At, 213 Po, 212 Po, 215 Po, or 177 Lu, and the imaging isotope 133 / 135 La, 133 Ce, 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 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 Figure 1 for an example. 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 of 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 18F) allows for a central production site covering most of the United States, and long-distance distribution of the resulting compound. Strictly speaking, with regard to radioactive decay, 212 Pb decays to alpha emitters 212 Bi, a beta emitter. 212 Pb is 212 Because the beta emissions resulting from the decay of Pb are physiologically of little importance compared to the alpha emissions, they are commonly referred to by physicians as alpha emitters. 212 After the Pb radionuclide emits beta emissions, 212 Pb 212 Bi (daughter product) remains in the chelator and becomes part of the therapeutic agent. 212 Bi is further decayed by one of two equivalent pathways (see Figure 1); (1) 212 Bi emits alpha emission 208 Tl and then emits beta-emission, or (2) 212 Bi emits beta emission 212 It becomes Po, stays in the chelator, and then immediately emits alpha emission. 212 Pb, and 212 Pb-containing drugs (including those described herein) showed a significant increase in alpha release after 11.65 hours ( 212 10.64 hours for Pb 212 Bi can be considered an alpha emitter with a physical half-life of 60.6 minutes (60.6 minutes for Bi). 212 The decay scheme of Pb (Fig. 1) is 208 When it becomes Pb, it produces one alpha emission and, incidentally, two beta emissions. This is of no consequence because the beta emission is about 10,000 times less massive than the alpha emission, and therefore the two beta emissions are insignificant compared to the alpha emission in terms of effects inside the body. When beta emitters are used for therapy, the total amount of radiopharmaceutical that needs to be injected to see an effect is orders of magnitude higher than the dose of a comparable alpha emitter.

[0150] 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 208 For 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 Pb) can be selected for chelation with two or more chelators. For therapeutic-only conjugates, radiotherapeutic isotopes (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 the 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., Cu) can be selected for chelation with two or more chelators. 64 Cu) and radiotherapeutic isotopes (e.g., 212 Pb) can be selected.

[0151] In some embodiments, a fluorescent dye is used instead of an imaging isotope. Non-limiting examples of fluorescent dyes include, for example, coumarin, cyanine, carboxyfluorescein, 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 (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 squara 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 benzophenone). 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); 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); dipyrromethene 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 nonspecifically (glutaraldehyde)).

[0152] For any conjugate, the binding moiety can be one or more small molecules, nanoparticles, liposomes, exosomes, polypeptides (e.g., antibodies or peptides), nucleic acids (e.g., DNA or RNA), oligonucleotides, viruses, or any other targeting biologics that bind to target molecules on cells (e.g., cancer cells). In some examples, the binding moiety is an extracellular vesicle, a virus, or glucose or a derivative thereof. In some examples, the binding moiety can target a molecule on the surface of a cell (e.g., a cell surface receptor). For example, a small molecule such as a Glu-ureido-based prostate-specific membrane antigen (PSMA) inhibitor (also known as a glutamate carboxypeptidase II inhibitor) can be used as the binding moiety. See, e.g., Kopka, et al., J. Nucl. Med., 58(Suppl. 2):17S-26S (2017). PSMA (also known as folate hydrolase 1 (FOLH1), FGCP, FOLH, GCP2, PSM, mGCP, GCPII, NAALAD1, or NAALAidase) is a cell membrane peptidase belonging to the M28B subfamily of the M28 peptidase family. For example, nanoparticles containing glutamate carboxypeptidase II inhibitors can be used as binding moieties. In some embodiments, the nanoparticles can be hydrophilic polyethylene glycol coronas bearing 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 as binding moieties.

[0153] For example, in some embodiments, the binding moiety can be a polypeptide that binds to PSMA, somatostatin receptor, fibroblast activation protein (FAP) polypeptide, melanocortin-1 receptor, B7-H3 protein, 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), 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), programmed death-ligand 1 (PD-L1) polypeptide, tumor-associated calcium signal transducer 2 (TROP2) polypeptide, protein tyrosine kinase 2 (PTK2) polypeptide, integrin beta 6 (ITGB6) polypeptide, 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 antibody fragment capable of binding to an antigen. As used herein, the term "antibody" 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.

[0154] 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 fragment exhibits the desired binding ability to a target molecule. An "Fv fragment" is the minimum 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 antigen-specific CDRs) still has the ability to recognize and bind antigen, although usually with lower affinity than the intact binding site. An "Fab fragment" is a fragment consisting of the constant domain of the light chain and the first constant domain of the heavy chain (C H1 "Fab fragments" also include the heavy chain C fragments, including one or more cysteines from the antibody hinge region. H1 It differs from "Fab' fragments" by the addition of a few residues at the carboxy terminus of the domain. "F(ab')2 fragments" are initially produced as a pair of "Fab' fragments" which have hinge cysteines between them. Such antibody fragments can be prepared by any suitable method, for example, papain or pepsin digestion.

[0155] In some examples, the antibody may be a humanized monoclonal antibody. Humanized monoclonal antibodies can be generated by introducing mouse complementarity-determining regions (CDRs) from the heavy and light variable chains of a mouse immunoglobulin into a human variable domain, followed by substituting human residues in the framework regions for their mouse counterparts. 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 are 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 instances, humanization, e.g., hyperhumanization, can be used as described in Hwang et al., Methods, 36:35-42 (2005).In some examples, 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 instances, 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).

[0156] Antibody fragments can be prepared by proteolytic hydrolysis of intact antibodies or by expression of nucleic acids encoding the fragment. Antibody fragments can be obtained by pepsin or papain digestion of intact antibodies by conventional methods. For example, Fab fragments can be generated by enzymatic cleavage of antibodies with papain. In some instances, antibody fragments can be generated by enzymatic cleavage of antibodies 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 generate a 3.5S Fab' monovalent fragment. In some instances, enzymatic cleavage using pepsin can be used to directly generate two monovalent Fab' fragments and an Fc fragment. These methods are described, for example, by Goldenberg (U.S. Pat. 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.

[0157] 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.

[0158] 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 NOs: 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 NOs: 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 NOs: 1-6, except that the variant polypeptide contains 1, 2, 3, 4, or 5 amino acid substitutions within the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 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 NOs: 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 NOs: 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).

[0159] [Table 1] TIFF2026502864000036.tif226148TIFF2026502864000037.tif90148

[0160] 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. Publ. No. 2014 / 0273078, European Patent Application Publication No. 3192810 A1, CN 108699157, European Patent No. 2,363,404, U.S. Pat. Appl. Publ. 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.

[0161] In some embodiments, the antibody that binds to PSMA can be the 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).

[0162] 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 the CDR amino acid sequence 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 can consist of the amino acid sequence set forth in any one of SEQ ID NOs: 7-12, except that the variant polypeptide contains 1, 2, 3, 4, or 5 amino acid substitutions within 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 can 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).

[0163] [Table 2] TIFF2026502864000039.tif34142

[0164] In some embodiments, the antibody that binds to the somatostatin receptor may be UMB1, UMB4, UMB5, or UMB7.

[0165] In some embodiments, the antibody that binds to the somatostatin receptor can be as described elsewhere, see, e.g., International Patent Application Publication No. WO 2018 / 005706, U.S. Patent Application Publication No. 2009 / 0016989, U.S. Patent Application Publication No. 2021 / 0340264, U.S. 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.

[0166] In some embodiments, the antibody can be an antibody that binds to a FAP polypeptide. For example, an antibody that binds to a FAP polypeptide can comprise a CDR that comprises, consists essentially of, or consists of the CDR amino acid sequence set forth in SEQ ID NOs: 13-18. In some examples, an antibody that binds to a FAP polypeptide can 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 to a FAP polypeptide can consist of the amino acid sequence set forth in any one of SEQ ID NOs: 13-18, except that the variant polypeptide contains 1, 2, 3, 4, or 5 amino acid substitutions within 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 can 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).

[0167] [Table 3]

[0168] In some embodiments, the antibody that binds to a FAP polypeptide can be sibrotuzumab or BMS168.

[0169] 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, U.S. Patent Application Publication No. 8,999,342, U.S. Patent Application Publication No. 2017 / 0369592, IL 281739 D0, U.S. Patent No. 9,481,730, and ES 2348556 T3.

[0170] 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 the 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 within 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 can be used in antibodies that bind to a CD3 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 4 (see also Example 17).

[0171] [Table 4] TIFF2026502864000042.tif75137

[0172] In some embodiments, the antibody that binds to a CD3 polypeptide can be muromonab or blinatumomab.

[0173] In some embodiments, an antibody that binds to 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.

[0174] 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 variant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions within 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 can 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).

[0175] [Table 5]

[0176] In some embodiments, the antibody that binds to a CD20 polypeptide can be tositumomab, tituximab, ofatumumab, obinutuzumab, ocrelizumab, or ublituximab.

[0177] In some embodiments, an antibody that binds to 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.

[0178] 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 the 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 variant polypeptide contains 1, 2, 3, 4, or 5 amino acid substitutions within 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 can be used in antibodies that bind to a CXCR4 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 6 (see also Example 17).

[0179] [Table 6] TIFF2026502864000045.tif221144TIFF2026502864000046.tif222144TIFF2026502864000047.tif50141

[0180] In some embodiments, the antibody that binds to a CXCR4 polypeptide can be ibalizumab, MAB172-100, PA3-305, or hz515H7.

[0181] In some embodiments, an antibody that binds to a CXCR4 polypeptide can be as described elsewhere. For example, EP 2285833 B1, JP 5749330 B2, U.S. Pat. No. 7,138,496, U.S. Pat. App. Pub. No. 2005 / 0002939, EP 2246364 A1, CA 2724409 A1, International Pat. App. Pub. No. WO 2006 / 089141, Broussas et al., Mol. Cancer Ther., 2016 Aug; 15(8):1890-9, International Pat. App. Pub. No. WO 2000 / 042074, International Pat. App. 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. App. Pub. No. WO See, for example, International Patent Application 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.

[0182] In some embodiments, the antibody can be an antibody that binds to a GRPR polypeptide.

[0183] In some embodiments, the antibody that binds to a GRPR polypeptide can be ABR-002, sc-398549, A30653.

[0184] In some embodiments, antibodies that bind to GRPR polypeptides 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.

[0185] 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 CDR 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 variant polypeptide contains 1, 2, 3, 4, or 5 amino acid substitutions within 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 can be used in antibodies that bind to a HER2 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 7 (see also Example 17).

[0186] [Table 7] TIFF2026502864000049.tif96136

[0187] In some embodiments, the antibody that binds to a HER2 polypeptide can be trastuzumab, pertuzumab, margetuximab, ZW25, or zumuzumab.

[0188] In some embodiments, the antibody that binds to the HER2 polypeptide can be as described elsewhere. For example, see 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.

[0189] In some embodiments, the antibody may be an antibody that binds to an MCR1 polypeptide.

[0190] In some embodiments, the antibody that binds to the MCR1 polypeptide may be ARC0638 or EPR6530.

[0191] 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 the 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 to a VEGF polypeptide may consist of the amino acid sequence set forth in any one of SEQ ID NOs: 43-48, except that the variant polypeptide comprises 1, 2, 3, 4, or 5 amino acid substitutions within 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 can be used in antibodies that bind to a VEGF polypeptide include, but are not limited to, the amino acid sequences set forth in Table 8 (see also Example 17).

[0192] [Table 8] TIFF2026502864000051.tif72137

[0193] In some embodiments, the antibody that binds to a VEGF polypeptide can be bevacizumab, ranibizumab, brolucizumab, or faricimab.

[0194] 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) of 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 contains one, two, three, four, or five amino acid substitutions within the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 345-350), 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 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 can 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).

[0195] [Table 9]

[0196] In some embodiments, the antibody that binds to a PD-L1 polypeptide may be atezolizumab, avelumab, durvalumab, BMS 936559, or cosibelimab.

[0197] 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 the CDR amino acid sequence 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 variant polypeptide contains one, two, three, four, or five amino acid substitutions within the linking sequence of the sequence identifier (e.g., any one of SEQ ID NOs: 351-356), 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: 351-356), 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: 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 can be used in antibodies that bind to a TROP2 polypeptide include, but are not limited to, the amino acid sequences set forth in Table 10 (see also Example 17).

[0198] [Table 10] TIFF2026502864000054.tif9138

[0199] In some embodiments, the antibody that binds to a TROP2 polypeptide may be sacituzumab or datopotamab.

[0200] In some embodiments, the binding moiety can be a liposome. The liposome can be an exosome, a vesicle, a nanovesicle, or an extracellular vesicle (EV). In some embodiments, the liposome can be derived from a mammalian cell, body fluid, or tissue. In some embodiments, the liposome can be derived from a primate. In some embodiments, the liposome can be derived from a human. The liposome can be derived from a cell, body fluid, or tissue, including, but not limited to, mesenchymal stem cells, adipocytes, T cells, dendritic cells, macrophages, mast cells, intestinal epithelial cells, B cells, neurons, oligodendrocytes, astrocytes, bone marrow cells, natural killer cells, plasma, serum, platelet cells, urine, lymphocytes, cerebrospinal fluid, cultured cells, milk, saliva, sweat, or ascites. The liposome can be preferentially targeted to tissues based on the expression of surface polypeptides. In some embodiments, integrin a6b4 and / or integrin a6b1 can target liposomes to cells in the lung. In some embodiments, integrin avb5 can target liposomes to cells in the liver. In some embodiments, tetraspanin Tspan8 and / or integrin a4 can target liposomes to cells in the pancreas. In some embodiments, integrin b4 can target liposomes to cells in the brain. In some embodiments, CD63 can target liposomes to neurons and glial cells. Liposomes can be selected for the natural presence of these polypeptides, or they can be engineered to have these polypeptides on their surface.

[0201] In some embodiments, the binding moiety may be a nucleic acid, hi some embodiments, the nucleic acid may be selected from one of thymine, adenine, guanine, cytosine, and uracil.

[0202] In some embodiments, the binding moiety can be an oligonucleotide. In some embodiments, the oligonucleotide can comprise DNA or RNA. In some embodiments, the oligonucleotide can be an aptamer. Targets to which an aptamer can bind include, but are not limited to, prostate-specific membrane antigen (PSMA), Erb-B2 receptor tyrosine kinase 2 (HER2), Erb-B2 receptor tyrosine kinase 3 (HER3), programmed cell death protein 1 (PD-1), programmed cell death 1 ligand 1 (PD-L1), vascular endothelial growth factor (VEGF), nucleolin, mucin 1, avβ3 integrin, CD49c, protein tyrosine kinase 7 (PTK7), tumor-associated calcium signal transduction protein 2 (TROP2), CD20, tenascin-C, carcinoembryonic antigen / immature laminin receptor protein (OFA / iLRP), periostin, platelet-derived growth factor (PDGF), or E2F transcription factor 3 (E2F3). The sequence of the aptamer used as the binding moiety can be as set forth in the table below. In some embodiments, the aptamer may be further modified with an inert polymer, hi some embodiments, the inert polymer is polyethylene glycol (PEG).

[0203] [Table 11] TIFF2026502864000056.tif239163TIFF2026502864000057.tif246162TIFF2026502864000058.tif241162 TIFF2026502864000059.tif228161TIFF2026502864000060.tif223162TIFF2026502864000061.tif245161

[0204] In some embodiments, the binding moiety can be a virus. The virus can be an adeno-associated virus (AAV), adenovirus, Maraba virus, measles virus, herpes simplex type 1 virus, lentivirus, vesicular stomatitis virus (VSV), Newcastle disease virus, myxoma virus, foamy virus, parvovirus, reovirus, or poliovirus. In some embodiments, the AAV can be AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12. In some embodiments, the AAV can have a hybrid capsid. In some embodiments, the AAV with a hybrid capsid can be AAV2 / 10, AAV2 / 11, AAVDJ, AAFDJ / 8, or AAV2i8. In some embodiments, the virus can be modified to express an additional peptide on its surface. In some embodiments, the binding moiety may be an engineered AAV, including but not limited to AAV9-PhP.B, AAV5-PhP.B, AAV-PHP.eB, AAV-PHP.Scan, AAV serotype Retrograde, or AAV serotype BR1, to be used to target brain malignancies. Furthermore, the above viral particles can also be PEGylated. Peptides include, but are not limited to, those listed in the table below.

[0205] [Table 12] TIFF2026502864000063.tif247161TIFF2026502864000064.tif36161

[0206] Exemplary Methods for Preparing Conjugates In some embodiments, the conjugate can be prepared according to methods and procedures similar to those described in PCT / US2022 / 034086, which is incorporated herein by reference. 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, 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 set forth 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 can be used as described herein include 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-hexahydrofuran, 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 the 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 linking moiety may also be 3-fluoro-2,2-dimethylpropionic acid or 2,2-dimethylpropionic acid.

[0207] As described herein, the conjugates provided herein can 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 can be capable of binding to one or more target molecules. For example, the binding moieties of the conjugates described herein can 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).

[0208] 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.

[0209] In some embodiments, one or more 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. In some embodiments, the cancer is a metastatic tumor (e.g., in a tissue selected from bone, liver, lung, brain, adrenal gland, etc.). In some embodiments, the metastatic tumor is about 1 mm, about 2 mm, about 3 mm, about 5 mm, about 10 mm, or 20 mm in size.

[0210] In some embodiments, one or more 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).

[0211] 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 several days to several 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 for an isotope used for imaging and one of the chelators being a chelator for an isotope used for radiotherapy, wherein the isotope used for imaging and the isotope used for radiotherapy are each complexed (chelated) to the chelator, and wherein the binding moieties bind to a tumor in the patient) can be formulated into a pharmaceutically 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, e.g., 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, causing them to decay at different rates.

[0212] 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 binds to a tumor in a patient, can be administered to the patient to determine the biodistribution of the conjugate (i.e., the location of the conjugate within 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 a chelator. Determining the biodistribution allows for personalized treatment doses for the patient and reduced side effects. Imaging can be performed after each administration of the conjugate to monitor treatment.

[0213] A therapeutically effective amount of the conjugates described herein can be formulated with one or more pharmaceutically acceptable carriers (additives or excipients) and / or diluents. In some embodiments, the additives stabilize against radiolysis. 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.

[0214] 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 phosphates, 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.

[0215] Pharmaceutical compositions containing one or more conjugates can be designed for oral or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, inhalation / aerosolized, intraarterial, intrathecal, intratumoral, intravesical, peritumoral, intraperitoneal, intracavity, and 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 presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and can be stored (e.g., in a freeze-dried (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.

[0216] In some examples, pharmaceutically acceptable compositions 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).

[0217] 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, or about 15 mCi to about 20 mCi). In some embodiments, an effective amount of a conjugate comprising a beta-emitting isotope is, 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 1,000 mCi, or about 1,000 mCi per cycle). The dose may be 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 600 mCi to about 1,400 mCi, about 700 mCi to about 1,300 mCi, about 800 mCi to about 1,200 mCi, or 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) is, 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 40 mCi, or about 40 mCi to about 40 mCi). about 30 mCi, about 4 mCi to about 35 mCi, about 4 mCi to about 35 mCi, about 5 mCi to about 40 mCi, about 5 mCi to about 35 mCi, about 5 mCi to about 30 mCi, about 5 mCi to about 25 mCi, about 5 mCi to about 20 mCi, about 10 mCi to about 30 mCi, about 15 mCi to about 40 mCi, about 20 mCi to about 40 mCi, or 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 1 mCi to about 9 ...1 mCi to about 95 mCi, about 2 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about 1 mCi to about 95 mCi, about The active ingredient may be about 5mCi to about 95mCi, about 3mCi to about 90mCi, about 4mCi to about 85mCi, about 4mCi to about 80mCi, about 5mCi to about 100mCi, about 5mCi to about 85mCi, about 5mCi to about 70mCi, about 5mCi to about 60mCi, about 5mCi to about 50mCi, about 10mCi to about 100mCi, about 15mCi to about 60mCi, about 20mCi to about 80mCi, or about 25mCi to about 100mCi).

[0218] In some embodiments in which more than one conjugate is administered, the effective amount of each conjugate may differ, e.g., 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.

[0219] 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 weighing approximately 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 dose and washout period is referred to as a treatment cycle.

[0220] If a particular mammal fails to respond to a particular amount of therapeutic agent conjugate, or if 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 twofold. 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, the frequency of administration, the duration of treatment, the use of multiple therapeutic agents, the route of administration, and the severity of the condition may require an increase or decrease in the actual effective amount administered.

[0221] The administration frequency 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 administration frequency of the conjugates can be about once daily, once a month, once every six weeks, once every two months, about once every three months, or about once every 16 weeks. The administration frequency of the conjugates described herein can remain constant throughout 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 16 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 can influence the actual frequency of administration used for a particular application, e.g., the effective amount, the duration of treatment, the use of multiple therapeutic agents, the route of administration, and the severity of the condition may require increased or decreased frequency of administration.

[0222] 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 several days to several months. Generally, 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. Multiple 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.

[0223] The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

[0224] Exemplary Conjugates for Targeting Cancer Suitable examples of radiometal-containing conjugates are provided in Figures 3A-6. In some cases, these structures may have various combinations of Cu-64, Cu-61, Cu-67, non-radioactive Cu, and Pb-212 / Pb-203 / Pb-non-radioactive. In some cases, these structures 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).

[0225] The following exemplary structures provide the conjugates prior to complexation with the metal(s).

[0226] [ka] TIFF2026502864000066.tif167137TIFF2026502864000067.tif83139TIFF20265028640 00068.tif152158TIFF2026502864000069.tif135160TIFF2026502864000070.tif215160

[0227] In the above conjugate, R 1 , R 2 , and R 3 are each independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH; R 1 , R 2 , and R 3 At least one of is other than C(=O)NH2 or C(=O)OH.

[0228] In the above conjugate, R 1 , R 2 , and R 3 At least one of is selected from: -S(=O)2OH, -S(=O)2NH2, -OP(=O)(OH)2, -P(=O)(OH)2, -OP(=O)(OH)NH2, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH2.

[0229] In the above conjugate, R 1 , R 2 , R 3 , and R 4 are each independently selected from C(=O)NH, C(=O)OH, -S(=O)OH, -S(=O)NH, -OP(=O)(OH), -P(=O)(OH), -OP(=O)(OH)NH, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH; R 1 , R 2 , R 3, and R 4 At least one of is other than C(=O)NH2 or C(=O)OH.

[0230] In the above conjugate, R 1 , R 2 , R 3 , and R 4 At least one of is selected from -S(=O)2OH, -S(=O)2NH2, -OP(=O)(OH)2, -P(=O)(OH)2, -OP(=O)(OH)NH2, -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH2.

[0231] In one example, R 1 is S(=O)2OH. In one example, R 1 is -S(=O)2NH2. In one example, R 1 is -OP(=O)(OH). In one example, R 1 is -P(=O)(OH)2. In one example, R 1 is -OP(=O)(OH)NH2. In one example, R 1 is C(=O)NHOH. In one example, R 1 is -C(=S)SH. In one example, R 1 is -C(=O)SH. In one example, R 1 is C(=S)OH. In one example, R 1 is C(=S)NH2.

[0232] In one example, R 2 is S(=O)2OH. In one example, R 2 is -S(=O)2NH2. In one example, R 2 is -OP(=O)(OH). In one example, R 2 is -P(=O)(OH)2. In one example, R 2 is -OP(=O)(OH)NH2. In one example, R 2 is C(=O)NHOH. In one example, R 2 is -C(=S)SH. In one example, R 2 is -C(=O)SH. In one example, R 2is C(=S)OH. In one example, R 2 is C(=S)NH2.

[0233] In one example, R 3 is S(=O)2OH. In one example, R 3 is -S(=O)2NH2. In one example, R 3 is -OP(=O)(OH). In one example, R 3 is -P(=O)(OH)2. In one example, R 3 is -OP(=O)(OH)NH2. In one example, R 3 is C(=O)NHOH. In one example, R 3 is -C(=S)SH. In one example, R 3 is -C(=O)SH. In one example, R 3 is C(=S)OH. In one example, R 3 is C(=S)NH2.

[0234] In one example, R 4 is S(=O)2OH. In one example, R 4 is -S(=O)2NH2. In one example, R 4 is -OP(=O)(OH). In one example, R 4 is -P(=O)(OH)2. In one example, R 4 is -OP(=O)(OH)NH2. In one example, R 4 is C(=O)NHOH. In one example, R 4 is -C(=S)SH. In one example, R 4 is -C(=O)SH. In one example, R 4 is C(=S)OH. In one example, R 4 is C(=S)NH2.

[0235] Additional examples of conjugates include:

[0236] [ka] TIFF2026502864000072.tif143143TIFF2026502864000073.tif104143

[0237] In some embodiments, exemplary conjugates include an imaging isotope complexed to an imaging isotope chelator and / or a radiotherapeutic isotope complexed to a radiotherapeutic isotope chelator. In some embodiments, the conjugate has the structure:

[0238] [ka] It has.

[0239] The binding moiety (targeting vector) in the conjugate herein may be selected from any of the polypeptides, antibodies, small molecules (e.g., glucose or a derivative thereof), extracellular vesicles (e.g., exosomes or liposomes), viruses, and nucleic acids described herein.

[0240] In some embodiments, the binding moiety is an adeno-associated virus. In some embodiments, the adeno-associated virus is AAV9.

[0241] In some embodiments, the binding moiety is an extracellular vesicle. In some embodiments, the extracellular vesicle is a plasma-derived extracellular vesicle.

[0242] The following are examples of glucose derivatives that can be used to target small metastatic tumors:

[0243] [ka]

[0244] Figures 6B, 7, and 8 depict conjugation reactions between alpha-PET platform moieties and extracellular vesicles, various types of viruses, and aptamers. In one example, these reactions are performed by conjugating a pre-radiolabeled alpha-PET moiety (labeled with an imaging isotope, a therapeutic isotope, or both) to a biologic. In another example, these conjugation reactions are performed without pre-radiolabeling the alpha-PET platform moiety, and the resulting conjugate is then radiolabeled via imaging and / or radiotherapy (if necessary). In one example of a conjugation reaction, an isothiocyanate group present on the alpha-PET platform moiety reacts with a primary amine-reactive group present on an extracellular vesicle, virus, or aptamer. To facilitate the conjugation reaction, the biologic reactant molecule can be modified with a linker containing an amino group. The conjugation reaction links the PET platform moiety and the biologic via a stable thiourea covalent bond. The biologics in the conjugates shown in Figures 6B, 7, and 8 are targeted vectors that allow for imaging and treatment of various diseases, including, but not limited to, various types of cancer. Exemplary conjugates are shown in Figures 6B, 7, and 8 for illustrative purposes only. Other biologics described herein can be conjugated to alpha-PET platform moieties using methods and procedures similar to those described in these figures.

[0245] The following table provides the amino acid sequences of exemplary binding moieties (targeting vectors) that may be used in the conjugates described herein.

[0246] [Table 13] TIFF2026502864000077.tif223154TIFF2026502864000078.tif183152

[0247] [Table 14] TIFF2026502864000080.tif216153TIFF2026502864000081.tif225152TIFF2026502864000082.tif116153

[0248] Table 15 TIFF2026502864000084.tif222153TIFF2026502864000085.tif223153TIFF2026502864000086.tif13152

[0249] Table 16 TIFF2026502864000088.tif225156TIFF2026502864000089.tif85154

[0250] Table 17 TIFF2026502864000091.tif170156

[0251] Table 18 TIFF2026502864000093.tif226152TIFF2026502864000094.tif226152TIFF2026502864000095.tif223153TIFF2026502864000096.tif29153

[0252] Table 19 TIFF2026502864000098.tif179153

[0253] Table 20 TIFF2026502864000100.tif225154TIFF2026502864000101.tif182155

[0254] [Table 21] TIFF2026502864000103.tif218154TIFF2026502864000104.tif137155

[0255] [Table 22] TIFF2026502864000106.tif136155 [Example]

[0256] Example 1 [ 64 Radiosynthesis and preclinical evaluation of [Cu]Cu-NOTA-TCMC-AAV [ 64 Cu]Cu-NOTA-TCMC-adenoassociated virus ([ 64 Radiosynthesis of [Cu]Cu-NOTA-TCMC-AAV was performed using AAV9 (Figure 9). For radiosynthesis, approximately 5.32 x 10 11AAV9 was first conjugated to SCN-TCMC-NOTA by incubating approximately 100 μL of AAV9 viral particles with approximately 50 μg of SCN-TCMC-NOTA (5 μL of 10 mg / mL SCN-TCMC-NOTA) in pH 9.0 phosphate-buffered saline (pH adjusted with 0.25 M Na2CO3) for approximately 16 hours at 37°C in a thermomixer shaking at 600 rpm. After incubation, NOTA-conjugated AAV9 particles were separated from unreacted SCN-TCMC-NOTA using a Sephadex G50 size-exclusion column. Purified NOTA-TCMC-AAV9 fractions after size-exclusion purification were identified by assessing the presence of AAV9 using quantitative polymerase chain reaction (qPCR) technology. The fractions with the highest number of AAV particles were identified. 64 Used for Cu chelation. 64 Chelation of Cu (approximately 0.37 mCi) was carried out in phosphate-buffered saline, pH 5.0 (pH adjusted with 10% phosphoric acid), for approximately 16 hours at room temperature. A chelation efficiency of approximately 85.4% was achieved as assessed by radio-TLC using a mobile phase of 100 mM sodium citrate, pH 5.0. In the radio-TLC chromatogram, the origin (Rf = approximately 0) was the intact [ 64 Cu]Cu-NOTA-TCMC-AAV9, and the solvent front (Rf = ca. 1) is unchelated. 64 Cu was expressed (Figure 10). After chelation, [ 64 Cu]Cu-NOTA-TCMC-AAV9 was isolated using a Sephadex G50 size exclusion column. 64 Cu was separated. Purified [ 64 Cu]Cu-NOTA-TCMC-AAV9 (approximately 86.2% radiochemical purity) was administered intravenously via tail vein injection in BALB / c mice, and PET images were acquired at various time points after injection (Figure 11). 64 Cu]Cu-NOTA-TCMC-AAV9 was distributed primarily to the liver after injection.

[0257] Example 2 [ 64Radiosynthesis and preclinical evaluation of Cu]Cu-NOTA-TCMC-EVs Figure 12 shows the 64 Cu]Cu-NOTA-TCMC-extracellular vesicles ([ 64 The chemical structure includes [Cu]Cu-NOTA-TCMC-EV). 64 Cu]Cu-NOTA-TCMC-extracellular vesicles ([ 64 Radiosynthesis of [Cu]Cu-NOTA-TCMC-EVs was performed using plasma-derived EVs (Figure 12). For radiosynthesis, plasma-derived EVs were first conjugated with SCN-TCMC-NOTA by incubating approximately 100 μg of EVs (approximately 100 μL) with approximately 100 μg of SCN-TCMC-NOTA (10 μL of 1 mg SCN-TCMC-NOTA / 0.1 mL HO) in pH 9.0 phosphate-buffered saline (pH adjusted with 0.25 M NaCO) for 60 minutes at 37°C in a thermomixer shaking at 600 rpm. After incubation, NOTA-conjugated EVs were separated from unreacted SCN-TCMC-NOTA using a Sephadex G50 resin size-exclusion column. Purified NOTA-TCMC-EV fractions after size-exclusion purification were identified by assessing the protein concentration in various fractions by Bradford assay. The fraction with the highest protein concentration was selected as the EV conjugated with SCN-TCMC-NOTA. 64 Used for Cu chelation. 64 Chelation of Cu (approximately 1.45 mCi) was carried out in phosphate-buffered saline, pH 5.0 (pH adjusted with 10% phosphoric acid) for 10 min at room temperature. Approximately 100% chelation efficiency was achieved as assessed by radio-TLC using a mobile phase of 100 mM sodium citrate, pH 5.0. In the radio-TLC chromatogram, the origin (Rf = approximately 0) was the intact [ 64 Cu]Cu-NOTA-TCMC-EV, and the solvent front (Rf = ca. 1) is non-chelated. 64 Cu was expressed (Figure 13). After chelation, [ 64 Cu]Cu-NOTA-TCMC-EV was isolated using a Sephadex G50 resin size exclusion column. 64 Cu was separated. Purified [64 Cu]Cu-NOTA-TCMC-EV (approximately 100% radiochemical purity) was administered intravenously via tail vein injection in BALB / c mice, and PET images were acquired at various time points after injection (Figure 14). 64 Cu]Cu-NOTA-TCMC-EVs were distributed mainly to the liver, heart, intestine, and bladder after injection.

[0258] Other embodiments While the present invention has been described with reference to its detailed description, it is understood that the foregoing description is intended to be illustrative, but 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 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; At least one chelator in the conjugate is C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH 2 , and C(=O)OR a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl; provided that all functional groups in the at least one chelator are C(=O)NH 2 or C(=O)OH; The chelator and the binding moiety are a moiety of formula (I): 【Chemistry 1】 (In the formula: Each X is N, P, P(=O), CR N and the moiety of formula (i): 【Chemistry 2】 are independently selected from x 1 , x 2 , x 3 , and x 4 each independently represents a point of attachment of the moiety of formula (I) to the chelator or the 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 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, wherein 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 the arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from the group consisting of OH, NO 2 ,CN,Haro,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 1 , y 2 , y 3 , and y 4 each is independently an integer from 1 to 10; Each R N is H, C 1-3 Alkyl, and C 1-3 haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5. The conjugate is attached via

2. The compound of formula (I) has the following formula: 【Transformation 3】 2. The conjugate of claim 1, having the formula:

3. The moiety of formula (I) is any one of the following formulas: 【Chemistry 4】 【change】 3. The conjugate of claim 2, having the formula:

4. The imaging isotope chelators were 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-hexaazabicyclo[6.6.6]eicosane-1,8-diazomethane (DTA), and 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC). amine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA), each of which is selected from the group consisting of C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH 2 , and C(=O)OR a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 The conjugate of any one of claims 1 to 3, wherein the heteroaryl is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

5. The imaging isotope chelator has at least one functional group C(=O)NH 2 or C(=O)NH, provided that it is other than C(=O)OH. 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 5. The conjugate of claim 4, substituted with three or four functional groups independently selected from:

6. The imaging isotope chelator has the formula: 【Transformation 5】 (In the formula, each R 1 , R 2 , and R 3 is C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 are independently selected from 【Transformation 6】 indicates the point of attachment to formula (I) 6. The conjugate of claim 4 or claim 5, having the formula:

7. R 1 , R 2 , and R 3 At least one of the following is C(=O)NH 2 or C(=O)OH.

8. the imaging isotope is: 133 Ce, 133 / 135 La, 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 The conjugate of any one of claims 1 to 7, wherein Y

9. said imaging isotope 64 The conjugate of claim 6, wherein the conjugate is Cu.

10. The radiotherapeutic isotope chelators were 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-hexaazabicyclo[6.6.6]eicosane-1,8-diazomethane (DTA), and 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC). amine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA), each of which is selected from the group consisting of C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, -C(=S)NH 2 , and C(=O)OR a wherein R a is C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 6-10 Aryl, C 3-10 The conjugate of any one of claims 1 to 9, wherein the heterocycloalkyl is selected from cycloalkyl, 5- to 10-membered heteroaryl, and 4- to 10-membered heterocycloalkyl.

11. The chelator for the radiotherapeutic isotope has at least one functional group C(=O)NH 2 or C(=O)NH, provided that it is other than C(=O)OH. 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 11. The conjugate of claim 10, substituted with 3 or 4 functional groups independently selected from:

12. The chelator of the radiotherapeutic isotope has the formula: 【Transformation 7】 (In the formula, each R 1 , R 2 , R 3 , and R 4 is C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 are independently selected from 【Transformation 8】 indicates the point of attachment to formula (I) 12. The conjugate of claim 10 or claim 11, having the formula:

13. R 1 , R 2 , R 3 , and R 4 At least one of the following is C(=O)NH 2 or C(=O)OH.

14. The conjugate of any one of claims 1 to 14, wherein the radiotherapeutic isotope is an alpha emitter.

15. The radiotherapeutic isotope is 223 / 225 / 227 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 The conjugate of any one of claims 1 to 14, wherein Lu

16. said radiotherapeutic isotope 212 16. The conjugate of claim 15, wherein the conjugate is Pb.

17. One of the following formulas: 【Chemistry 9】 【change】 (In the formula, each R 1 , R 2 , R 3 , and R 4 is C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 are independently selected from R 1 , R 2 , R 3 , and R 4 At least one of the following is C(=O)NH 2 or other than C(=O)OH) The conjugate of claim 1, selected from:

18. The conjugate of any one of claims 1 to 17, wherein the binding moiety is selected from a polypeptide, an antibody, a small molecule, an extracellular vesicle, a virus, and a nucleic acid.

19. 19. The conjugate of claim 18, wherein the polypeptide binds to a prostate-specific membrane antigen, a somatostatin receptor, or a melanocortin-1 receptor.

20. 19. The conjugate of claim 18, wherein the small molecule is a glutamate carboxypeptidase II inhibitor.

21. 19. The conjugate of claim 18, wherein the small molecule is glucose or a derivative thereof.

22. The conjugate is any one of the following compounds: 【Chemistry 10】 【change】 【change】 【change】 (In the formula, each R 1 , R 2 , R 3 , and R 4 is C(=O)NH 2 , C(=O)OH, -S(=O) 2 OH, -S(=O) 2 NH 2 , -OP(=O)(OH) 2 , -P(=O)(OH) 2 , -OP(=O)(OH)NH 2 , -C(=O)NHOH, -C(=S)SH, -C(=O)SH, -C(=S)OH, and -C(=S)NH 2 are independently selected from R 1 , R 2 , R 3 , and R 4 At least one of the following is C(=O)NH 2 or other than C(=O)OH) The conjugate of claim 1, selected from:

23. 23. A method of treating cancer in a mammal in need thereof, said method comprising administering to said mammal a conjugate of any one of claims 1 to 22, said conjugate comprising said imaging isotope complexed to said chelator of said imaging isotope, and said conjugate comprising said radiotherapeutic isotope complexed to said chelator of said radiotherapeutic isotope.

24. 1. A method of treating cancer in a mammal, comprising: a) administering to said mammal a first conjugate according to any one of claims 1 to 22, 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 the mammal an amount of a second conjugate according to any one of claims 1 to 22, which is identical to the first conjugate except that it comprises the radiotherapeutic isotope complexed to the chelator of the radiotherapeutic isotope. The method comprising:

25. 25. The method of claim 24, wherein 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.

26. 26. The method of any one of claims 23 to 25, wherein the cancer is selected from the group consisting of prostate cancer, neuroendocrine cancer, colon cancer, lung cancer, pancreatic cancer, melanoma, and lymphatic cancer.

27. 27. The method of claim 26, wherein the cancer is a metastatic tumor.

28. 23. A method of treating cancer in a mammal in need thereof, said method comprising administering to said mammal two or more conjugates of any one of claims 1 to 22, wherein each conjugate comprises two or more chelators and a binding moiety, one of said chelators being a chelator for an imaging isotope and one of said 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 radiotherapeutic isotope; The method.

29. 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; the binding moiety is selected from an extracellular vesicle, a virus, a nucleic acid, and glucose or a derivative thereof; The chelator and the binding moiety are a moiety of formula (I): 【Chemistry 11】 (In the formula: Each X is N, P, P(=O), CR N and the moiety of formula (i): 【Chemistry 12】 are independently selected from x 1 , x 2 , x 3 , and x 4 each independently represents a point of attachment of the moiety of formula (I) to the chelator or the 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 3-10 Cycloalkylene, C 6-10 arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene, wherein 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 the arylene, 5- to 14-membered heteroarylene, and 4- to 10-membered heterocycloalkylene is selected from the group consisting of OH, NO 2 ,CN,Haro,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 1 , y 2 , y 3 , and y 4 each is independently an integer from 1 to 10; Each R N is H, C 1-3 Alkyl, and C 1-3 haloalkyl; n is an integer selected from 1, 2, 3, 4, and 5. The conjugate is attached via

30. 30. The conjugate of claim 29, wherein the binding moiety is an extracellular vesicle.

31. 30. The conjugate of claim 29, wherein the binding moiety is a virus.

32. 30. The conjugate of claim 29, wherein the binding moiety is a nucleic acid.

33. The compound of formula (I) has the following formula: 【Chemistry 13】 The conjugate of any one of claims 29 to 32, having the formula:

34. The moiety of formula (I) is any one of the following formulas: 【Chemistry 14】 【change】 34. The conjugate of claim 33, having the formula:

35. The imaging isotope chelators were independently selected from 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)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] The conjugate of any one of claims 29 to 34, comprising a compound selected from the group consisting of eicosane-1,8-diamine (DiAmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA).

36. The radiotherapeutic isotope chelators were 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-hexaazabicyclo[6.6.6]eicosane-1,8-diamine (Di 36. The conjugate of any one of claims 29 to 35, comprising a compound selected from the group consisting of AmSar), N,N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), deferoxamine (DFO), diethylenetriaminepentaacetic acid (DTPA), and N,N'-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18crown-6 (MACROPA).

37. 37. The conjugate of any one of claims 29 to 36, wherein the chelator for the imaging isotope is 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) and the chelator for the radiotherapeutic isotope is 1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetracyclododecane (TCMC).

38. the imaging isotope is: 133 / 135 La, 133 Ce, 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 The conjugate of any one of claims 29 to 37, wherein Y

39. said imaging isotope 64 39. The conjugate of claim 38, wherein Cu.

40. 40. The conjugate of any one of claims 29 to 39, wherein the radiotherapeutic isotope is an alpha emitter.

41. The radiotherapeutic isotope is 223 / 225 / 227 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 The conjugate of any one of claims 29 to 40, wherein Lu

42. said radiotherapeutic isotope 212 42. The conjugate of claim 41, wherein the conjugate is Pb.

43. Any one of the following compounds: 【Chemistry 15】 【change】 【change】 30. The conjugate of claim 29, selected from:

44. 10. A method of treating cancer in a mammal in need thereof, said method comprising administering to said mammal a conjugate of any one of claims 29 to 43, said conjugate comprising said imaging isotope complexed to said chelator of said imaging isotope, and said conjugate comprising said radiotherapeutic isotope complexed to said chelator of said radiotherapeutic isotope.

45. 1. A method of treating cancer in a mammal, comprising: a) administering to said mammal a first conjugate according to any one of claims 29 to 43, 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 the mammal an amount of a second conjugate according to any one of claims 26 to 40, which is identical to the first conjugate except that it comprises the radiotherapeutic isotope complexed to the chelator of the radiotherapeutic isotope. The method comprising:

46. 46. ​​The method of claim 45, wherein 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.

47. 47. The method of any one of claims 44 to 46, wherein the cancer is selected from the group consisting of prostate cancer, neuroendocrine cancer, colon cancer, lung cancer, pancreatic cancer, melanoma, and lymphatic cancer.

48. 48. The method of claim 47, wherein the cancer is a metastatic tumor.

49. 10. A method of treating cancer in a mammal in need thereof, said method comprising administering to said mammal two or more conjugates of any one of claims 26 to 40, wherein each conjugate comprises two or more chelators and a binding moiety, one of said chelators being a chelator for an imaging isotope and one of said 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 radiotherapeutic isotope; The method.

50. 30. The conjugate of claim 29, wherein the binding moiety is glucose or a derivative thereof.

51. The conjugate is 【Chemistry 16】 and the binding moiety is an adeno-associated virus.

52. 52. The conjugate of claim 51, wherein the adeno-associated virus is AAV9.

53. The conjugate is 【Chemistry 17】 and the binding moiety is an extracellular vesicle.

54. 54. The conjugate of claim 53, wherein the extracellular vesicles are plasma-derived extracellular vesicles.

55. 23. The conjugate of any one of claims 1 to 22, wherein the conjugate comprises an imaging isotope complexed to said chelator of said imaging isotope and / or wherein the conjugate comprises a radiotherapeutic isotope complexed to said chelator of said radiotherapeutic isotope.

56. The conjugate is [Chemistry 18] 54. The conjugate of claim 53, wherein:

57. 57. The conjugate of claim 56, wherein the binding moiety is an adeno-associated virus.

58. 58. The conjugate of claim 57, wherein the adeno-associated virus is AAV9.

59. 57. The conjugate of claim 56, wherein the binding moiety is an extracellular vesicle.

60. 60. The conjugate of claim 59, wherein the extracellular vesicles are plasma-derived extracellular vesicles.

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