DOTA-hapten compositions for pre-targeted radioimmunotherapy using anti-DOTA / antitumor antigen bispecific antibodies

Abstract

The present disclosure provides compositions and methods for the detection and treatment of cancer. In particular, the compositions of the present technology include novel compounds that can be complexed with radioisotopes. Methods of using the DOTA-haptens of the present technology in diagnostic imaging and pre-targeted radioimmunotherapy are also disclosed herein.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims the benefits and priority of U.S. Provisional Patent Application No. 63 / 045,632, filed on June 29, 2020, the content of which is hereby incorporated by reference in its entirety. Technical Field The present technology generally relates to compositions containing novel DOTA - haptens, and methods of using them in diagnostic imaging and pre - targeted radioimmunotherapy. Description of Government Support This invention was made with government support under CA008748 and CA184746 awarded by the National Institutes of Health. The government has certain rights in this invention.

Background Art

[0002] The following description of the background of the present technology is provided merely to assist in the understanding of the present technology and is not recognized as describing or constituting the prior art of the present technology. Radiolabeled drugs have been used for over 50 years as delivery vehicles for ionizing radiation to specific disease sites (Larson SM. Cancer 67:1253-1260 (1991); Britton KE. Nucl Med Commun. 18:992-1007 (1997)). Numerous molecules, including radiolabeled antibodies, antibody fragments, alternative scaffolds, and small molecules, have been investigated for targeted delivery of radioisotopes (Tolmachev V, et al. Cancer Res. 67:2773-2782 (2007); Birchler MT, et al., Otolaryngol Head Neck Surg. 136:543-548 (2007); Reubi JC, Maecke HR. J Nucl Med. 49:1735-1738 (2008)). The use of antibodies to target toxins to tumors, such as radioimmunotherapy (RIT) with direct conjugate antibodies, has been problematic, for one thing, due to suboptimal tumor dose and therapeutic index (TI). Furthermore, dose escalation is not feasible due to normal tissue bystander toxicity, and therefore such treatments produce limited antitumor effects. On top of that, antibodies exhibit a long blood half-life and result in a low tumor-to-background ratio. Antibody fragments and other smaller binding scaffolds exhibit more rapid blood clearance but result in high renal and / or hepatic uptake. Radiolabeled small molecule ligands generally exhibit more rapid blood clearance and lower background compared to antibodies and antibody fragments, but usually result in insufficient specificity due to their relatively low affinity for the desired target. In pre-targeted radioimmunotherapy (PRIT), a non-radioactive bifunctional antibody that is specific for both a tumor antigen and a small hapten is administered and localized to the tumor. After sufficient blood clearance of the antibody, a radiolabeled small molecule is administered and captured by the pre-targeted antibody. However, many small peptides and metal chelate haptens used in PRIT systems exhibit significant systemic retention, which results in unwanted background radioactivity that limits the signal-to-background ratio of imaging and contributes to non-specific radiation that limits the maximum tolerated dose for therapeutic applications (Orcutt et al., Mol Imaging Biol 13:215-221 (2011)). SUMMARY OF THE INVENTION

[0003] Therefore, there is a need for novel molecules that enable (a) efficient pre-targeted radioimmunotherapy of tumors in vivo and (b) rapid clearance of radiolabeled small molecules from non-tumor tissues.

[0004] SUMMARY OF THE TECHNOLOGY In one aspect, the present disclosure provides a compound of formula I

Chemical formula

[0005] In another aspect, the present disclosure provides a bischelate comprising any of the above compounds of formula I and a radionuclide cation. In some embodiments, the bischelate is of formula II

Chemical formula

[0006] In all and all embodiments, the compound of formula II is R 2 It contains radioactive nuclide cations chelated by a group. The radioactive nuclide cations may be alpha-emitting isotopes, beta-emitting isotopes, Auger emitters, or a combination of two or more of these. An example of an alpha-emitting isotope is: 213 Bi, 211 At, 225 Ac, 152 Dy, 212 Bi, 223 Ra, 219 Rn, 215 Po, 211 Bi, 221 Fr, 217 At, and 255 This includes, but is not limited to, Fm. Examples of beta particle-emitting isotopes are: 86 Y, 90 Y, 89 Sr, 165Dy, 186 Re, 188 Re, 177 Lu, and 67 Examples of Auger emitters include, but are not limited to, Cu. 111 In, 67 Ga, 51 Cr, 58 Co, 99m Tc, 103m Rh, 195m Pt, 119 Sb, 161 Ho, 189m Os, 192 Ir, 201 Tl, and 203 Contains Pb. In some embodiments of the compound of formula II, the radioactive nuclide cation is 89 Zr, 68 Ga, 203 Pb, 212 Pb, 227 Th, or 64 It is Cu.

[0007] In some embodiments, radioactive nuclide cations have decay energies in the range of 20 to 6,000 keV. The decay energies may be in the range of 60 to 200 keV for Auger emitters, 100 to 2,500 keV for beta emitters, and 4,000 to 6,000 keV for alpha emitters. The maximum decay energy of useful beta particle-emitting nuclides may be in the range of 20 to 5,000 keV, 100 to 4,000 keV, or 500 to 2,500 keV. The decay energy of useful Auger emitters may be <1,000 keV, <100 keV, or <70 keV. The decay energy of useful alpha particle-emitting radionuclides may be in the range of 2,000 to 10,000 keV, 3,000 to 8,000 keV, or 4,000 to 7,000 keV.

[0008] In another embodiment, the Disclosure provides a complex comprising a compound of formula I and a bispecific antibody that recognizes and binds to the compound and a tumor antigen target. The Disclosure also provides a complex comprising a bischelate of formula II and a bispecific antibody that binds to the bischelate and a tumor antigen target. In any of the above embodiments of the complexes disclosed herein, the bispecific antibody may be an infinite binder. In some embodiments, the bispecific antibody comprises an antigen-binding fragment of C825 (see Cheal et al., Mol Cancer Ther. 13(7):1803-12 (2014)) or 2D12.5 (Corneillie et al., J. Inorganic Biochemistry 100:882-890 (2006)). Additionally or alternatively, in any of the above embodiments of the complexes disclosed herein, the bispecific antibody comprises an antigen-binding fragment of C825 having a G54C substitution. Additionally or alternatively, in any of the above embodiments of the complex disclosed herein, the bispecific antibody comprises an antigen-binding fragment of 2D12.5 having a G54C substitution.

[0009] In any of the embodiments of the complex disclosed herein, the tumor antigen target is GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase (acetylglucosaminyltrans ferase)V intron V sequence), prostate cancer psm, PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, lung resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le yThe antigen is selected from the group consisting of E-cadherin, V-cadherin, and EpCAM. In addition or alternatively, in some embodiments of the complex, the bispecific antibody is added to the compound or bischelate in the following concentrations: 100nM-95nM, 95-90nM, 90-85nM, 85-80nM, 80-75nM, 75-70nM, 70-65nM, 65-60nM, 60-55nM, 55-50nM, 50-45nM, 45-40nM, 40-35nM, 35-30nM, 30-25nM, 25-20nM, 20-15nM, 15-10nM, 10-5nM, 5-1nM, 1nM-950pM, 950pM-900pM, 900pM-850pM, 850pM-800pM, 800pM-75 0pM, 750pM~700pM, 700pM~650pM, 650pM~600pM, 600pM~550pM, 550pM~500pM, 50 0pM~450pM, 450pM~400pM, 400pM~350pM, 350pM~300pM, 300pM~250pM, 250pM~20 K levels of 0 pM, 200 pM to 150 pM, 150 pM to 100 pM, 100 pM to 50 pM, 50 pM to 40 pM, 40 pM to 30 pM, 30 pM to 20 pM, 20 pM to 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2.5 pM, 2 pM, 1.5 pM, or less than 1 pM. d They are joined together.

[0010] In one embodiment, the present disclosure provides a method for detecting a tumor in a subject requiring such detection, comprising: (a) administering an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody bound to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the complex; and (b) detecting the presence of a tumor in the subject by detecting a radioactivity level released by the complex that is higher than a reference value. The tumor may be a solid tumor or a humoral tumor. In some embodiments, the subject is a human.

[0011] In another embodiment, the Disclosure provides a method for selecting a subject for pre-targeted radioimmunotherapy, comprising: (a) administering an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody bound to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the complex; (b) detecting the level of radioactivity released by the complex; and (c) selecting the subject for pre-targeted radioimmunotherapy if the level of radioactivity released by the complex is higher than a reference value. In some embodiments, the subject is a human.

[0012] In some embodiments of the methods disclosed herein, the radioactivity level emitted by the complex is detected using positron emission tomography or single-photon emission computed tomography. Additionally or alternatively, in some embodiments of the methods disclosed herein, the subject is diagnosed with or suspected of having cancer. Cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. In some embodiments, brain cancer is pituitary adenoma, meningioma, neuroblastoma, or craniopharyngioma. In addition or alternatively, in some embodiments of the methods disclosed herein, the complex is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. In certain embodiments, the complex is administered into the cerebrospinal fluid or blood of the subject.

[0013] In some embodiments of the methods disclosed herein, the radioactivity level released by the complex is detected between 4 and 24 hours after administration of the complex. In certain embodiments of the methods disclosed herein, the radioactivity level released by the complex is expressed as a percentage of the injected dose per gram of tissue (%ID / g). In some embodiments, the ratio of radioactivity levels between tumor tissue and normal tissue is approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:1. In another embodiment, the Disclosure provides a method for enhancing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, comprising the steps of (a) administering to the subject an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target, and (b) administering to the subject an effective amount of a bischelate of formula II configured to bind to the anti-DOTA bispecific antibody. In some embodiments, the method further comprises administering to the subject an effective amount of a clearing agent prior to the administration of the bischelate. The clearing agent may be a 500kD aminodextran-DOTA conjugate (e.g., 500kD dextran-DOTA-Bn(Y), 500kD dextran-DOTA-Bn(Lu), or 500kD dextran-DOTA-Bn(In)). In some embodiments, the subject is human.

[0014] Additionally or alternatively, in some embodiments of the method, the tumor antigen targets are GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence) ), prostate cancer PSM, PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, lung resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le y The antigen is selected from the group consisting of E-cadherin, V-cadherin, and EpCAM. In addition or alternatively, in some embodiments of the method, the anti-DOTA bispecific antibody and / or bischelate is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally.

[0015] In one embodiment, the present disclosure provides a method for enhancing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, comprising the steps of administering an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody that recognizes and binds to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing the tumor antigen target recognized by the bispecific antibody of the complex. The complex may be administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. In some embodiments, the subject is human.

[0016] In another aspect, the Disclosure provides a method for treating cancer in a subject where it is needed, comprising the steps of (a) administering to the subject an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target, and (b) administering to the subject an effective amount of a bischelate of formula II configured to bind to the anti-DOTA bispecific antibody. In a particular embodiment, the method further comprises administering to the subject an effective amount of a clearing agent prior to the administration of the bischelate. Also provided herein is a method for treating cancer in a subject where it is needed, comprising the steps of administering to the subject an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody that recognizes and binds to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the complex.

[0017] A method for treating cancer may further include the step of sequentially, separately, or simultaneously administering at least one chemotherapeutic agent selected from the group consisting of nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosourea, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxin, platinum coordination complexes, vinca alkaloids, substituted ureas, methylhydrazine derivatives, corticosteroids, hormone antagonists, endostatins, taxol, camptothecin, SN-38, doxorubicin, doxorubicin analogs, antimetabolites, alkylating agents, antimitotic agents, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, apoptosis promoters, methotrexate, and CPT-11. In some embodiments, cancer is selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. In some embodiments, the subject is human.

[0018] Kits containing components suitable for treating or diagnosing cancer in patients are also disclosed herein. In one embodiment, the kit comprises a compound or bischelate of the Technology, at least one anti-DOTA bispecific antibody, and instructions for use. The kit may further comprise a clearing agent (e.g., 500 kDa aminodextran conjugated to DOTA) and / or one or more radionuclides. [Brief explanation of the drawing]

[0019] [Figure 1A]This figure shows a plot of percentage injection dose per gram (%ID / g) versus time for mice injected with the compound of this technology, which contains the chelated radionuclide ([203Pb]TCMC-PEG4-LuDOTA). These results demonstrate that the majority (>97%) of [203Pb]TCMC-PEG4-LuDOTA is eliminated from the plasma after 1 hour. [Figure 1B] This figure shows the calibration curve for Pb-203 in the gamma counter window (150-500 keV). [Figure 2] This figure shows the ex vivo in vivo distribution studies of 89Zr activity for pre-targeted [89Zr]DFO-PEG4-LuDOTA and untargeted [89Zr]DFO-PEG4-LuDOTA in various tissues of SW1222- tumor-bearing and tumor-free mice, 4 hours (h) after injection (pi). Data are presented as mean ± standard deviation. [Figure 3] This figure shows whole blood sampling via retroorbital hemorrhage in tumor-free mice administered [89Zr]DFO-PEG4-LuDOTA. Data are shown as mean ± standard deviation. IA / g% refers to the percentage area under the ideal dose-volume histogram curve (IA) per gram. [Figure 4] This figure shows the systemic 89Zr activity in tumor-free mice administered with [89Zr]DFO-PEG4-LuDOTA. Data are shown as mean ± standard deviation. [Figure 5] This figure shows representative PET maximum intensity projection images from two different mice that underwent PRIT using [89Zr]DFO-PEG4-LuDOTA (200 pmol / 1.48 MBq). The images were obtained 4 hours after injection of [89Zr]DFO-PEG4-LuDOTA. The signal was detected in the scGPA33-expressing SW1222 xenograft (circled area). [Figure 6]This figure shows the ex vivo in vivo distribution study of 68Ga activity for pre-targeted [68Ga]NODAGA-PEG4-LuDOTA and [68Ga]DO3A-PEG4-LuDOTA (described in WO2019 / 010299) in various tissues of SW1222-tumor-bearing mice 1 hour (h) after injection (pi). Data are shown as mean ± standard deviation. For mol calculations, the doses prepared were 225 μCi and 145 μCi for [68Ga]NODAGA-PEG4-LuDOTA and [68Ga]DO3A-PEG4-LuDOTA, respectively. *Outliers at 5.02% ID / g are not excluded (2.44 ± 2.30). [Figure 7] This figure shows a representative PET image (coronal section) of a mouse that underwent PRIT using [68Ga]NODAGA-PEG4-LuDOTA (130 pmol / 6.0 MBq). The image was obtained 1 hour after injection of [68Ga]NODAGA-PEG4-LuDOTA. The tumor is clearly visible in the shoulder region ("T"). [Figure 8] This figure shows a series of ex vivo in vivo studies of 68Ga activity related to pre-targeted [68Ga]NODAGA-PEG4-LuDOTA in various tissues of SW1222-tumor-bearing mice. Data are presented as mean ± standard deviation. *Outliers 0.428 ± 0.299 g excluded, except for 0.0631 g; **Outliers 6.68 ± 3.49 IA / g% excluded, except for 0.0631 g. [Figure 9] Figure 8 shows the 68Ga activity time curves for tumors, blood, and kidneys, based on a series of ex vivo in vivo distribution data collected at 5, 15, 30, and 60 minutes after injection of pre-targeted [68Ga]NODAGA-PEG4-LuDOTA. The data in the graph are shown as mean ± standard deviation. [Figure 10]This figure shows the ex vivo in vivo distribution study of 64Cu activity related to pre-targeted [64Cu]NODAGA-PEG4-LuDOTA in various tissues of SW1222-tumor-bearing mice 24 hours after injection. Data are shown as mean ± standard deviation. *Outliers at 1.92% ID / g are not excluded (0.63 ± 0.86); **Outliers at 0.18% ID / g are not excluded (0.06 ± 0.08). [Figure 11] This figure shows a representative PET image (coronal section) of a mouse that underwent PRIT using [64Cu]NODAGA-PEG4-LuDOTA. The image was obtained approximately 24 hours after injection of 300 μCurie [64Cu]NODAGA-PEG4-LuDOTA. The tumor is clearly visible in the shoulder region ("T"). [Figure 12A-B] This figure shows the ex vivo in vivo distribution study of 177Lu activity for pre-targeted [177Lu]DOTABn-PEG4-LuDOTA (also known as "[177Lu]Lu-GeminiDOTA") in various tissues of SW1222-tumor-bearing mice 24 hours after injection. Data are shown as % injectable activity per gram of tissue (IA / g%), (mean ± SEM). [Figure 13] This figure shows the ex vivo in vivo distribution study of 203Pb activity for pre-targeted [203Pb]TCMC-PEG4-LuDOTA (also referred to herein as "[203Pb]TCMC-Proteus-DOTA") or [203Pb]DO3A-PEG4-LuDOTA (also referred to herein as "[203Pb]Proteus-DOTA") in various tissues of SW1222-tumor-bearing mice 24 hours after injection. Data are shown as % injectable activity per gram of tissue (IA / g%), (mean ± SD). [Figure 14]This figure shows the ex vivo in vivo distribution studies of 111In activity for pre-targeted [111In]Proteus-DOTA(Lu) or [111In]Proteus-DOTA(Gd) in various tissues of SW1222-tumor-bearing mice 24 hours after injection. Data are shown as % injectable activity per gram of tissue (IA / g%), (mean ± SD). [Modes for carrying out the invention]

[0020] It should be noted that certain aspects, methods, embodiments, variations, and features of this method are described below with varying degrees of detail in order to provide a substantial understanding of the Art.

[0021] In carrying out this method, many conventional techniques in molecular biology, protein biochemistry, cell biology, microbiology, and recombinant DNA are used. For example, Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; the series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., NY); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach;Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual;Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition;Gait ed. (1984) Oligonucleotide Synthesis;US Patent No. 4,683,195;Hames and Higgins eds. (1984) Nucleic Acid Hybridization;Anderson (1999) Nucleic Acid Hybridization;Hames and Higgins eds. (1984) Transcription and Translation;Immobilized Cells and Enzymes (IRL Press (1986));Perbal (1984) A Practical Guide to Molecular Cloning;Miller and Calos eds.See (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); and Herzenberg et al. eds. (1996) Weir's Handbook of Experimental Immunology.

[0022] The composition of this technology comprises a novel DOTA-hapten useful in diagnostic imaging / dosimetry and PRIT (e.g., alpha particle radioimmunotherapy). The DOTA-PRIT platform is accompanied by a three-step pre-targeting strategy comprising: (1) an IgG-single-chain variable fragment (scFv) bispecific antibody construct (IgG-scFv) containing an antibody sequence of an anti-DOTA-hapten single-chain variable fragment scFv "C825" with an antitumor antigen antibody (IgG portion) and pM affinity; (2) a 500kD dextran-DOTA-hapten clearing agent; and (3) administration of the radiolabeled DOTA-hapten composition of this technology.

[0023] Previous studies have used anti-GPA33-DOTA-PRIT, respectively, for theranostic beta-particle radioimmunotherapy (RIT) or in vivo positron emission tomography (PET) of athymic nude mice carrying GPA33-expressing colon cancer xenografts. 177 Lu- or 86 We have demonstrated that YS-2-(4-aminobenzyl)-1,4,7,10-tetraazacyclododecanetetraacetate chelate (DOTA-Bn) hapten can be pre-targeted. However, in vivo using the model PRIT system... 225 Pre-targeting with Ac-DOTA-Bn takes place 24 hours after injection. 225Ac-DOTA-Bn showed little tumor uptake (<1% ID / g). See WO2019 / 010299. Therefore, conventional DOTA-haptens, 225 It is not suitable for DOTA-PRIT radiotherapy applications involving high-energy-transferring (LET) alpha-emitting isotopes such as Ac.

[0024] In contrast, the compositions disclosed herein (a) enable efficient in vivo pre-targeted radiotherapy of tumors, (b) exhibit complete renal clearance without unwanted renal / systemic retention, and (c) can bind with high affinity to anti-DOTA bispecific antibodies (e.g., anti-huA33-C825) (i.e., the DOTA hapten compositions of this technology do not sterically block the interaction between the lutetium-DOTA moiety of the DOTA hapten composition and the anti-DOTA bispecific antibody).

[0025] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as generally understood by those skilled in the art to which this art belongs. Where used herein and in the appended claims, the singular forms “a,” “an,” and “the” include multiple references unless explicitly indicated otherwise. For example, a reference to “a cell” includes a combination of two or more cells. Generally, the nomenclature used herein, as well as the laboratory procedures in cell culture, molecular genetics, organic chemistry, analytical chemistry, nucleic acid chemistry, and hybridization described below, are well known and commonly used in the art. As used herein, the term “about” with respect to numbers is generally interpreted, unless otherwise stated or the context makes it clear, to include numbers that fall within 1%, 5%, or 10% of the number in either direction (greater than or less than) (except where such numbers fall less than 0% or greater than 100% of the possible value).

[0026] The phrase "and / or" as used in this disclosure will be understood to mean any one of the individually listed members or any combination of two or more of them—for example, "A, B, and / or C" will mean "A, B, C, A and B, A and C, or B and C." pharmaceutically acceptable salts of the compounds described herein include acid or base addition salts that are within the scope of this technology, retain the desired pharmacological activity, and are not biologically undesirable (e.g., the salts are not excessively toxic, allergenic, or irritating, and are biologically usable). If the compounds of this technology have a basic group, such as an amino group, pharmaceutically acceptable salts can be formed with inorganic acids (e.g., hydrochloric acid, hydroboric acid, nitric acid, sulfuric acid, and phosphoric acid), organic acids (e.g., alginates, formic acid, acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid, tartaric acid, lactic acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and p-toluenesulfonic acid) or acidic amino acids (e.g., aspartic acid and glutamic acid). If the compounds of this technology have an acidic group, such as a carboxylic acid group, this can be formed with metals such as alkalis and alkaline earth metals (e.g., Na + Li + , K + Ca 2+ Mg 2+ Zn 2+ ), ammonia, or organic amines (e.g., dicyclohexylamine, trimethylamine, triethylamine, pyridine, picoline, ethanolamine, diethanolamine, triethanolamine) or basic amino acids (e.g., arginine, lysine, and ornithine) can form salts. Such salts can be prepared in situ during the isolation and purification of the compound, or by reacting the purified compound separately with a suitable acid or base in its free base or free acid form, respectively, and then isolating the salts thus formed.

[0027] As used herein, “administration” of a drug or substance to a subject includes any route through which the compound is introduced or delivered to the subject to perform its intended function. Administration may be carried out by any preferred route, for example, orally, intranasally, parenterally (intravenously, intramuscularly, intraperitoneally, or subcutaneously), rectally, or topically. Administration may include self-administration and administration by another person.

[0028] As used herein, the term “antibody” collectively refers to immunoglobulins or immunoglobulin-like molecules, including, but not limited to, IgA, IgD, IgE, IgG, and IgM, combinations thereof, and similar molecules produced during an immune response in any vertebrate, such as mammals (e.g., humans, goats, rabbits, and mice) and non-mammalian species (e.g., shark immunoglobulins). As used herein, “antibody” (including “intact immunoglobulin”) and “antigen-binding fragment” specifically bind to the molecule of interest (or a group of highly similar molecules of interest) with substantially no binding to other molecules (e.g., a binding constant for the molecule of interest that is about 10 times higher than the binding constant for other molecules in a biological sample). 3 M -1 Twice as large, about 10 4 M -1 Twice as large or about 10 5 M -1 Antibodies and antibody fragments having a binding constant twice as large. The term "antibody" also includes genetically modified forms such as chimeric antibodies (e.g., humanized mouse antibodies) and heteroconjugate antibodies (e.g., bispecific antibodies). Pierce Catalog and Handbook, 1994-1995 (Pierce Chemical Co., Rockford, Ill.); Kuby, J., Immunology, 3 rd See also Ed., WH Freeman & Co., New York, 1997.

[0029] More specifically, an antibody refers to a polypeptide ligand that specifically recognizes and binds to an antigen epitope and contains at least a light-chain immunoglobulin variable region or a heavy-chain immunoglobulin variable region. Antibodies are composed of a heavy chain and a light chain, which each contains a variable heavy chain (V H ) region and variable light chain (V L It has a variable region called the ) region. In addition, V H Region and V L The region is responsible for the binding of antigens recognized by antibodies. Typically, immunoglobulins have heavy (H) and light (L) chains interconnected by disulfide bonds. There are two types of light chains: lambda (λ) and kappa (κ). There are five main heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgM, IgD, IgG, IgA, and IgE. Each heavy and light chain contains a constant region and a variable region (the region is also known as a "domain"). Together, the heavy and light chain variable regions bind specifically to the antigen. The light and heavy chain variable regions contain a "framework" region interrupted by three hypervariable regions, also called "complementarity-determining regions" or "CDRs." The extent of the framework regions and CDRs is defined (see Kabat et al., Sequences of Proteins of Immunological Interest, US Department of Health and Human Services, 1991, incorporated herein by reference). The Kabat database is currently maintained online. The sequences of different light chain or heavy chain framework regions are relatively conserved within a species. The framework region of an antibody, i.e., the combined framework region of its constituent light and heavy chains, mostly adopts a β-sheet conformation, and the CDRs connect the β-sheet structures, forming loops that in some cases form part of the β-sheet structure. Thus, the framework region acts to form a scaffold that allows the CDRs to be positioned in the correct orientation through non-covalent interactions between the chains.

[0030] CDRs are primarily responsible for binding to the antigen's epitope. The CDRs on each chain are typically numbered sequentially from the N-terminus as CDR1, CDR2, and CDR3, and are also typically identified by the chain on which a particular CDR is located. Therefore, V H CDR3 is located in the variable domain of the heavy chain of the antibody in which it is present, whereas V L CDR1 is a CDR1 derived from the variable domain of the light chain of the antibody in which it is present. Antibodies that bind to a target protein or molecule (e.g., DOTA) have specific V H Region and V L Antibodies have a regional sequence and therefore a specific CDR sequence. Antibodies with different specificities (i.e., different combination sites for different antigens) have different CDRs. It is the CDR that changes depending on the antibody, but only a limited number of amino acid positions within the CDR are directly involved in antigen binding. These positions within the CDR are called specificity-determining residues (SDRs). Examples of antibodies include monoclonal antibodies, polyclonal antibodies, humanized antibodies, chimeric antibodies, recombinant antibodies, multispecific antibodies, bispecific antibodies, and antibody fragments. Antibodies bind specifically to antigens.

[0031] A "bispecific antibody" is an antibody that can simultaneously bind to two different antigens. A bispecific antibody (BsAb) and a bispecific antibody fragment (BsFab) may have, for example, at least one arm that specifically binds to a tumor-associated antigen, and at least one other arm that specifically binds to a targetable conjugate carrying a therapeutic or diagnostic agent (e.g., the bischelate of this technology). A variety of different bispecific antibody structures are known in the art. In some embodiments, each binding site in a bispecific antibody is derived from a different monoclonal antibody. H and / or V L Includes region. In some embodiments, the bispecific antibody contains a CDR derived from the first monoclonal antibody. H and / or V L V contains an immunoglobulin molecule having a region, and a CDR derived from a second monoclonal antibody. H and / or V LIt includes antibody fragments having a region (e.g., Fab, F(ab'), F(ab')2, Fd, Fv, dAB, scFv, etc.).

[0032] As used herein, the term “diabody” means a small antibody fragment having two antigen-binding sites, wherein the light chain variable domain (V) in the same polypeptide chain is the same. L ) connected to the heavy chain variable domain (V H )(V H V L This refers to a fragment containing ). By using a linker that is too short to allow pairing between two domains on the same chain, those domains are forced to pair with complementary domains on another chain, creating two antigen-binding sites. Diabodies are described in more detail, for example, EP404,097;WO93 / 11161; and 30 Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993).

[0033] As used herein, the terms "single-chain antibody" or "single-chain Fv (scFv)" refer to the two domains of the Fv fragment, V L and V H This refers to an antibody fusion molecule. A single-chain antibody molecule may contain a polymer containing several individual molecules, such as a dimer, trimer, or other polymer. Furthermore, F v Two domains of the fragment, V L and V H It is encoded by a separate gene, but using a recombination method, V L and V H The regions pair up to form a monovalent molecule (single-stranded F). v (scF vThey can be linked by synthetic linkers, which allow them to be constructed as a single protein chain forming what is known as a single-chain antibody (BAT). Bird et al. (1988) Science 242:423-426 and Huston et al. (1988) Proc. Natl. Acad Sci. USA 85:5879-5883. Such single-chain antibodies can be prepared by recombinant techniques or by enzymatic or chemical cleavage of intact antibodies.

[0034] As used herein, the terms “intact antibody” or “intact immunoglobulin” mean an antibody or immunoglobulin having at least two heavy (H) chain polypeptides and two light (L) chain polypeptides interconnected by disulfide bonds. Each heavy chain has a heavy chain variable region (HCVR or V) H It consists of a heavy chain constant region (abbreviated as CH1, CH2, and CH3). Each light chain is composed of a light chain variable region (LCVR or V in this specification). L It consists of a (abbreviated as) and a light chain steady region. The light chain steady region is one domain, C L It consists of V. H and V L The region can be further subdivided into a highly variable region called the Complementarity Determination Region (CDR), which incorporates a more conserved region called the Framework Region (FR). H and V L It consists of three CDRs and four FRs, arranged from the amino terminus to the carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of the antibody can mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

[0035] As used herein, “antigen” refers to a molecule to which an antibody can selectively bind. A target antigen may be a protein (e.g., an antigenic peptide), a carbohydrate, a nucleic acid, a lipid, a hapten, or another naturally occurring or synthetic compound. Antigens may also be administered to animal subjects to induce an immune response in them. As used herein, the term “antigen-binding fragment” refers to a fragment of a total immunoglobulin structure that contains a portion of the polypeptide responsible for binding to an antigen. Examples of antigen-binding fragments useful in this technology include scFv, (scFv)2, scFvFc, Fab, Fab' and F(ab')2, diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0036] "Binding affinity" refers to the strength of the combined non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). The affinity of molecule X for its partner Y is generally expressed by the dissociation constant (K). d Affinity can be expressed by ( ). Affinity can be measured by standard methods known in the art, including those described herein. Low-affinity conjugates generally contain antibodies that tend to dissociate easily from the antigen, while high-affinity conjugates generally contain antibodies that tend to remain bound to the antigen for a longer duration.

[0037] As used herein, “clearing agent” is a drug that binds to excess bifunctional antibodies present in the target blood compartment, facilitating rapid clearance by the kidney. The use of clearing agents before hapten administration facilitates a better tumor-to-background ratio in the PRIT system. Examples of clearing agents include 500kD dextran-DOTA-Bn(Y) (Orcutt et al., Mol Cancer Ther. 11(6): 1365-1372 (2012)), 500kD aminodextran-DOTA conjugates, and antibodies against pre-targeted antibodies. As used herein, “control” refers to an alternative sample used for comparative purposes in an experiment. A control may be “positive” or “negative.” For example, if the purpose of an experiment is to determine the correlation of the efficacy of a therapeutic agent for treating a particular type of disease or condition, a positive control (a compound or composition known to exhibit the desired therapeutic effect) and a negative control (a subject or sample that receives no treatment or a placebo) are typically used.

[0038] As used herein, the term “effective amount” of a composition means an amount sufficient to achieve the desired preventive or therapeutic effect, for example, an amount that causes a reduction in symptoms associated with the disease being treated, e.g., a disease or medical condition related to the target polypeptide (e.g., breast cancer, colorectal cancer, brain cancer, etc.). The amount of the composition of this technology administered to a subject depends on the degree, type and severity of the disease, as well as individual characteristics, e.g., overall health, age, sex, weight and drug tolerance. Those skilled in the art can determine an appropriate dosage according to these and other factors. The composition of this technology may also be administered in combination with one or more additional therapeutic compounds. As used herein, the term “epitope” means an antigenic determinant capable of specific binding to an antibody. Epitopes typically consist of chemically active surface groups of a molecule and usually possess specific three-dimensional structural features and specific charge characteristics. As used herein, “infinite binder” refers to an anti-metal chelate bispecific antibody characterized by the formation of a highly specific permanent bond between the bispecific antibody and the metal chelate upon binding. See Corneillie et al., J. Inorganic Biochemistry 100:882-890 (2006).

[0039] As used herein, the term “sample” refers to a clinical sample obtained from a subject or isolated microorganism. In certain embodiments, a sample is obtained from a biological source such as tissue, body fluid, or microorganism taken from a subject (i.e., a “biological sample”). Sample sources include, but are not limited to, mucus, sputum, bronchoalveolar lavage fluid (BAL), bronchial lavage fluid (BW), whole blood, body fluid, cerebrospinal fluid (CSF), urine, plasma, serum, or tissue. As used herein, the term “separate” therapeutic use means administering at least two active ingredients simultaneously or substantially simultaneously via different routes. As used herein, the term “sequential” therapeutic use means administering at least two active ingredients at different times, either via the same or different routes of administration. More specifically, sequential use means administering one active ingredient entirely before administration of the other active ingredient begins. Thus, one active ingredient may be administered over several minutes, hours, or days before administration of the other active ingredient. In this case, there is no simultaneous treatment. As used herein, the term “concurrent” therapeutic use means administering at least two active ingredients via the same route, and simultaneously or substantially simultaneously.

[0040] As used herein, “specifically bind” means that a molecule (e.g., an antibody) recognizes and binds to another molecule (e.g., an antigen), but substantially does not recognize and bind to other molecules. As used herein, “specifically binds,” “specifically binds to” a particular molecule (e.g., an antigen, or an epitope on an antigen), or “specific to” means, for example, about 10 times the amount of the molecule it binds to. -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, or 10 -12 M's K dIt can be represented by molecules that possess it. As used herein, the terms “subject,” “individual,” or “patient” are used interchangeably and refer to individual organisms, vertebrates, mammals, or humans. In certain embodiments, the individual, patient, or subject is a human. As used herein, the term “therapeutic agent” is intended to mean a compound that, when present in an effective amount, produces a desired therapeutic effect on a subject in need.

[0041] As used herein, “to treat” or “treatment” encompasses treatment of a disease or disorder described herein in a subject, e.g., a human, and includes (i) inhibiting the disease or disorder, i.e., preventing its onset; (ii) mitigating the disease or disorder, i.e., causing regression of the disorder; (iii) slowing the progression of the disorder; and / or (iv) inhibiting, mitigating, or slowing the progression of one or more symptoms of the disease or disorder. “To treat cancer” means that symptoms associated with cancer are, for example, alleviated, reduced, cured, or placed in remission. It should also be recognized that the various treatment methods for diseases described herein are intended to mean “substantial” treatments, including both complete and incomplete treatments, in which some biological or medically relevant outcome is achieved. Treatment may be a continuous, long-term treatment for a chronic disease, or a single or multiple administration for the treatment of an acute condition.

[0042] A "tautomer" refers to an isomer of a compound that is in equilibrium with itself. The presence and concentration of these isomers depend on the environment in which the compound is found; for example, they may differ depending on whether the compound is in a solid state, an organic solution, or an aqueous solution. For example, in an aqueous solution, quinazolinone may exhibit the following isomers, which are referred to as tautomers of each other.

[0043] [ka] As another example, guanidine may exhibit the following isomeric forms in protic organic solutions (e.g., water), which are also referred to as tautomers of each other. [ka] Because there are limitations to representing compounds using structural formulas, please understand that all chemical formulas of the compounds described herein represent all tautomers of the compound and are within the scope of this technology. The stereoisomers (also known as optical isomers) of a compound include all chiral, diastereomer, and racemic forms of the structure unless a specific stereochemistry is explicitly indicated. Therefore, the compounds used in this technique, as is evident from the expression, contain concentrated or decomposed optical isomers at any or all chiral atoms. Both racemic and diastereomer mixtures, as well as individual optical isomers, can be isolated or synthesized such that they substantially do not contain their enantiomer or diastereomer pairs, and all of these stereoisomers are within the scope of this technique.

[0044] The compounds of this technology may exist as solvates, particularly hydrates. Hydrates may form during the manufacture of the compound or a composition containing the compound, or they may form over time due to the hygroscopic properties of the compound. Similarly, the compounds of this technology may exist as organic solvent hydrates, including, in particular, DMF, ethers, and alcohol solvates. The identification and preparation of any specific solvate is within the scope of the art of those skilled in synthetic organic chemistry or medicinal chemistry.

[0045] Pre-targeted radioimmunotherapy (PRIT) Pre-targeting is a multi-step process that addresses the slow blood clearance of tumor-targeting antibodies, which contributes to undesirable toxicity to normal tissues such as bone marrow. In pre-targeting, a radionuclide or other diagnostic or therapeutic agent is attached to a low-molecular-weight hapten. A pre-targeted bispecific antibody with binding sites for the hapten and the target antigen is administered first. Then, the unbound antibody is cleared from the circulation, and subsequently, the hapten is administered. DOTA-PRIT is used to effectively target beta-emitting radioactive isotopes (e.g., lutetium-177) to GD2 or GPA33-expressing human cancer xenografts, thereby reducing toxicity to normal tissues such as bone marrow and kidney. Beta particle emission (e.g., 177 Lu-DOTA-Bn hapten is considered to be a low-energy conferral with energies in the range of 1-10 nm and 0.1-1 MeV. DOTA-PRIT is a beta-particle emitting radioactive isotope of lutetium and yttrium (respectively, 177 Lu and 90 It is optimally suited for targeting Y) because anti-DOTA C825 (anti-DOTA scFv) binds to DOTA complexes containing such radioactive lanthanides with an affinity of pM.

[0046] However, solid tumors are generally radioresistant. On the other hand, alpha particle radiotherapy (for example, 225 Using Ac-DOTA-hapten, alpha-particle radiotherapy delivers highly linearly energized alpha particles with an energy range of 50-80 μm and 5-8 MeV, resulting in highly potent cytotoxic activity with minimal secondary damage. Unlike beta particles, which can deliver their energy over longer distances, alpha-particle radiotherapy has high therapeutic potential for small-volume tumors, including minimal residual disease, which can be a major cause of cancer recurrence. Therefore, it is necessary to enhance the effectiveness of DOTA-PRIT radiotherapy using alpha-particle emitters, which have greater therapeutic potential compared to beta particles.

[0047] A unique limitation of C825 is the variation in the binding affinity that scFv has for various anti-DOTA-haptens, which is highly dependent on the ionic radius of the trivalent rare earth element. Previous modeling studies have demonstrated that a hapten binding affinity of 100 pM is required for efficient delivery of ionizing radiation in PRIT (assuming conditions of high antigen density and saturated BsAb dose), specifically to achieve near-maximal hapten retention in angiotumors and micrometastases. C825 is used with DOTA-Bn[S-2-(4-aminobenzyl)-1,4,7,10-tetraazacyclododecanetetraacetic acid chelate] complexes of Y, Lu, or Gd, with 15.4±2.0 pM, 10.8±2.5 pM, or 34.0±5.3 pM of K, respectively. d It was shown that binding occurs at (equilibrium dissociation constant, mean ± SD). In contrast, K for DOTA-Bn complexes containing In or Ga d The values ​​were 1.01±0.04 nM or 52±12 nM. Therefore, DOTA-PRIT is well-suited for targeting beta particle emitters such as yttrium-90 and lutetium-177, but less likely to be suitable for alpha particle emitters (e.g., actinium isotopes).

[0048] In preliminary experiments, a model DOTA-PRIT system (anti-GD2-DOTA-PRIT) was used in vivo. 225 When pre-targeted using Ac-DOTA-Bn, the equimolar dose was 177 Compared to Lu-DOTA-Bn, 24 hours after injection 225 Statistically significant (p ≤ 0.005; unpaired two-sided Student's t-test) and insignificant intratumoral uptake (as %ID / g; mean ± standard deviation (SD)) of Ac-DOTA-Bn; 225 For Ac-DOTA-Bn (n=5): 0.82±0.17; 177 Regarding Lu-DOTA-Bn (n=5): It has been shown that a result of 10.29±2.87 was obtained. See WO2019 / 010299. No significant differences were observed in normal tissues such as blood or kidneys (regarding blood: 225 Ac- or 177For Lu-DOTA-Bn, the values ​​were 0.33±0.03 or 0.49±0.09, respectively; for the kidney: 225 Ac- or 177 For Lu-DOTA-Bn, the results were 0.65±0.15 or 0.83±0.10, respectively (both p>0.05). This suggests that the in vivo results for the two tracers were similar, and that in vivo stability was likely not a limiting factor for tumor localization.

[0049] Composition of this technology DOTA is a macrocyclic chelating agent that forms stable metal complexes that are irreversible under physiological conditions. DOTA has a molecular weight of 405 daltons and exhibits rapid diffusion and renal clearance. DOTA and its variants chelate a wide range of metals, including paramagnetic metals and radionuclides. Exemplary metals include yttrium, indium, gallium, gadolinium, europium, terbium, lutetium, copper, bismuth, actinium, and all lanthanide metals.

[0050] In one embodiment, the present disclosure relates to a compound of formula I. [ka] (I) or a pharmaceutically acceptable salt thereof (wherein M 1 Chelate 175 Lu 3+ , 45 Sc 3+ , 69 Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ ,151 EU 3+ , 153 EU 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ , or 160 Gd 3+ and; R 1 teeth, [ka] [ka] [ka] [ka] and X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X31 , X 32 , X 33 , X 34 , X 35 , and X 36 Each is independently either a lone pair of electrons (i.e., giving the oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 Each is independently S or O; Q 1 n is S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22. In a particular embodiment, n is 3.

[0051] In another embodiment, the present disclosure provides a bischelate comprising any of the above compounds of formula I and a radionuclide cation. In some embodiments, the compound of formula I contains about 1 pM to 1 nM (e.g., about 1 to 10 pM; 1 to 100 pM; 5 to 50 pM; 100 to 500 pM; or 500 pM to 1 nM) of K in the radionuclide cation. d It can be joined by K d The bischelate is in the range of approximately 1 nM to approximately 1 pM, for example, approximately 1 nM, 950 pM, 900 pM, 850 pM, 800 pM, 750 pM, 700 pM, 650 pM, 600 pM, 550 pM, 500 pM, 450 pM, 400 pM, 350 pM, 300 pM, 250 pM, 200 pM, 150 pM, 100 pM, 90 pM, 80 pM, 70 pM, 60 pM, 50 pM, 40 pM, 30 pM, 20 pM, 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2.5 pM, 2 pM, or 1 pM or less. In some embodiments, the bischelate is of formula II.

[0052] [ka] (II) or a pharmaceutically acceptable salt thereof (wherein M 1 Chelate 175 Lu 3+ , 45 Sc 3+ , 69 Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ , 151 EU 3+ , 153 EU 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ , or 160 Gd 3+ and; R 2 teeth, [ka] [ka] [ka] [ka] and; M 2 Each instance is independent of R 2 A radioactive nuclide cation chelated by a group; X1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , and X 36 Each is independently either a lone pair of electrons (i.e., giving the oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 Each is independently S or O; Q 1 n is S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22. In certain embodiments, n is 3. In some embodiments, as an addition or alternative, the radioactive nuclide cation is a divalent or trivalent cation.

[0053] In all and all embodiments, the compound of formula II is R 2It contains radioactive nuclide cations chelated by a group. The radioactive nuclide cations may be alpha-emitting isotopes, beta-emitting isotopes, Auger emitters, or a combination of two or more of these. An example of an alpha-emitting isotope is: 213 Bi, 211 At, 225 Ac, 152 Dy, 212 Bi, 223 Ra, 219 Rn, 215 Po, 211 Bi, 221 Fr, 217 At, and 255 This includes, but is not limited to, Fm. Examples of beta particle-emitting isotopes are: 86 Y, 90 Y, 89 Sr, 165 Dy, 186 Re, 188 Re, 177 Lu, and 67 Examples of Auger emitters include, but are not limited to, Cu. 111 In, 67 Ga, 51 Cr, 58 Co, 99m Tc, 103m Rh, 195m Pt, 119 Sb, 161 Ho, 189m Os, 192 Ir, 201 Tl, and 203 Contains Pb. In some embodiments of the compound of formula II, the radioactive nuclide cation is 89 Zr, 68 Ga, 203 Pb, 212 Pb, 227 Th, or 64 It is Cu.

[0054] In some embodiments, radioactive nuclide cations have decay energies in the range of 20 to 6,000 keV. Decay energies may be in the range of 60 to 200 keV for Auger emitters, 100 to 2,500 keV for beta emitters, and 4,000 to 6,000 keV for alpha emitters. The maximum decay energy of useful beta particle-emitting nuclides may be in the range of 20 to 5,000 keV, 100 to 4,000 keV, or 500 to 2,500 keV. The decay energy of useful Auger emitters may be <1,000 keV, <100 keV, or <70 keV. The decay energy of useful alpha particle-emitting radionuclides may be in the range of 2,000 to 10,000 keV, 3,000 to 8,000 keV, or 4,000 to 7,000 keV.

[0055] In another embodiment, the Disclosure provides a complex comprising a compound of formula I and a bispecific antibody that recognizes and binds to the compound and a tumor antigen target. The Disclosure also provides a complex comprising a bischelate of formula II and a bispecific antibody that binds to the bischelate and a tumor antigen target. In any of the above embodiments of the complexes disclosed herein, the bispecific antibody may be an infinite binder. In some embodiments, the bispecific antibody comprises an antigen-binding fragment of C825 (see Cheal et al., Mol Cancer Ther. 13(7):1803-12 (2014)) or 2D12.5 (Corneillie et al., J. Inorganic Biochemistry 100:882-890 (2006)). Additionally or alternatively, in any of the above embodiments of the complexes disclosed herein, the bispecific antibody comprises an antigen-binding fragment of C825 having a G54C substitution. Additionally or alternatively, in any of the above embodiments of the complex disclosed herein, the bispecific antibody comprises an antigen-binding fragment of 2D12.5 having a G54C substitution.

[0056] In any of the embodiments of the complex disclosed herein, the tumor antigen target is GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase (acetylglucosaminyltrans ferase)V intron V sequence), prostate cancer psm, PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, lung resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le yThe antigen is selected from the group consisting of E-cadherin, V-cadherin, and EpCAM. In addition or alternatively, in some embodiments of the complex, the bispecific antibody is added to the compound or bischelate in the following concentrations: 100nM-95nM, 95-90nM, 90-85nM, 85-80nM, 80-75nM, 75-70nM, 70-65nM, 65-60nM, 60-55nM, 55-50nM, 50-45nM, 45-40nM, 40-35nM, 35-30nM, 30-25nM, 25-20nM, 20-15nM, 15-10nM, 10-5nM, 5-1nM, 1nM-950pM, 950pM-900pM, 900pM-850pM, 850pM-800pM, 800pM-75 0pM, 750pM~700pM, 700pM~650pM, 650pM~600pM, 600pM~550pM, 550pM~500pM, 50 0pM~450pM, 450pM~400pM, 400pM~350pM, 350pM~300pM, 300pM~250pM, 250pM~20 K levels of 0 pM, 200 pM to 150 pM, 150 pM to 100 pM, 100 pM to 50 pM, 50 pM to 40 pM, 40 pM to 30 pM, 30 pM to 20 pM, 20 pM to 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2.5 pM, 2 pM, 1.5 pM, or less than 1 pM. d They are joined together.

[0057] Diagnostic and treatment methods of this technology In one embodiment, the present disclosure provides a method for detecting a tumor in a subject where it is needed, comprising the steps of: (a) administering an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody conjugated to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the complex; and (b) detecting the presence of a tumor in the subject by detecting a radioactivity level released by the complex that is higher than a reference value. Also disclosed herein is a method for detecting a tumor in a subject where it is needed, comprising the steps of: (a) administering an effective amount of an anti-DOTA bispecific antibody to a subject, wherein the anti-DOTA bispecific antibody is configured to localize to a tumor expressing a tumor antigen target; (b) administering an effective amount of a bischelate of formula II to a subject, wherein the bischelate is configured to conjugate to the anti-DOTA bispecific antibody; and (c) detecting the presence of a tumor in the subject by detecting a radioactivity level released by the bischelate that is higher than a reference value. The anti-DOTA bispecific antibody is administered under conditions and for a duration sufficient to saturate tumor cells (e.g., according to a drug regimen). In some embodiments, the unbound anti-DOTA bispecific antibody is removed from the bloodstream after administration of the anti-DOTA bispecific antibody. In some embodiments, the bischelate of formula II is administered after a period that may be sufficient to allow clearance of the unbound anti-DOTA bispecific antibody. In addition or alternative, in some embodiments of the methods disclosed herein, the tumor is a solid tumor or a humoral tumor. In all and all embodiments of the methods disclosed herein, the step of detecting the tumor in a subject includes the step of imaging the tumor in vivo and / or measuring the amount or dose of radiation absorbed by the subject. In some embodiments, the subject is human.

[0058] In another aspect, the Disclosure provides a method for selecting a subject for pre-targeted radioimmunotherapy, comprising: (a) administering an effective amount of a conjugate comprising a bischelate of formula II and a bispecific antibody bound to the bischelate and a tumor antigen target, wherein the conjugate is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the conjugate; (b) detecting the level of radioactivity released by the conjugate; and (c) selecting the subject for pre-targeted radioimmunotherapy if the level of radioactivity released by the conjugate is higher than a reference value. A method for selecting a target for pre-targeted radioimmunotherapy is also provided herein, comprising: (a) administering an effective amount of an anti-DOTA bispecific antibody to the target, wherein the anti-DOTA bispecific antibody is configured to localize to a tumor expressing a tumor antigen target; (b) administering an effective amount of a bischelate of formula II to the target, wherein the bischelate is configured to bind to the anti-DOTA bispecific antibody; (c) detecting the level of radioactivity released by the bischelate; and (d) selecting a target for pre-targeted radioimmunotherapy if the level of radioactivity released by the bischelate is higher than a reference value. In addition or alternatively, in some embodiments of the methods disclosed herein, the tumor is a solid tumor or a humoral tumor. In some embodiments, the subject is a human.

[0059] In some embodiments of the methods disclosed herein, the radioactivity level emitted by the complex is detected using positron emission tomography or single-photon emission computed tomography. Additionally or alternatively, in some embodiments of the methods disclosed herein, the subject is diagnosed with or suspected of having cancer. Cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. In some embodiments, brain cancer is pituitary adenoma, meningioma, neuroblastoma, or craniopharyngioma. In some embodiments, brain cancer is a pituitary adenoma, meningioma, neuroblastoma, or craniopharyngioma. In addition or alternatively, in some embodiments of the methods disclosed herein, the complex is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. In certain embodiments, the complex is administered into the cerebrospinal fluid or blood of the subject.

[0060] In some embodiments of the methods disclosed herein, the radioactivity level released by the complex is detected between 4 and 24 hours after administration of the complex. In certain embodiments of the methods disclosed herein, the radioactivity level released by the complex is expressed as a percentage of the injected dose per gram of tissue (%ID / g). A baseline value may be calculated by measuring the radioactivity level present in non-tumor (normal) tissue and calculating the mean radioactivity level ± standard deviation present in the non-tumor (normal) tissue. In some embodiments, the baseline value is the standard uptake value (SUV). See Thie JA, J Nucl Med. 45(9):1431-4 (2004). In some embodiments, the ratio of radioactivity levels between tumor tissue and normal tissue is approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:1.

[0061] In another embodiment, the present disclosure provides a method for enhancing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, comprising the steps of (a) administering an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target to the subject, and (b) administering an effective amount of a bischelate of formula II configured to bind to the anti-DOTA bispecific antibody to the subject. In some embodiments, the subject is human. The anti-DOTA bispecific antibody is administered under conditions and for a sufficient period of time (e.g., according to a drug regimen) sufficient to saturate the tumor cells. In some embodiments, after administration of the anti-DOTA bispecific antibody, the unbound anti-DOTA bispecific antibody is removed from the bloodstream. In some embodiments, the bischelate of formula II is administered after a period of time that may be sufficient to allow clearance of the unbound anti-DOTA bispecific antibody.

[0062] Bischelate can be administered at any point between 1 minute and 4 days or more after administration of the anti-DOTA bispecific antibody. For example, in some embodiments, the bischelate is administered at 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, or any range within that range. Alternatively, bischelate may be administered at any point at least 4 days after administration of the anti-DOTA bispecific antibody.

[0063] In addition or alternatively, in some embodiments, the method further includes the step of administering an effective amount of a clearing agent to the target before administering the bischelate. The clearing agent may be any molecule (dextran, dendrimer, or polymer) that can be conjugated with C825-hapten. In some embodiments, the clearing agent is 2000kD, 1500kD, 1000kD, 900kD, 800kD, 700kD, 600kD, 500kD, 400kD, 300kD, 200kD, 100kD, 90kD, 80kD, 70kD, 60kD, 50kD, 40kD, 30kD, 20kD, 10kD, or 5kD or less. In some embodiments, the clearing agent is a 500kD aminodextran-DOTA conjugate (e.g., 500kD dextran-DOTA-Bn(Y), 500kD dextran-DOTA-Bn(Lu), or 500kD dextran-DOTA-Bn(In)).

[0064] In some embodiments, the clearing agent and the bischelate of formula II are administered without further administration of the anti-DOTA bispecific antibody. For example, in some embodiments, the anti-DOTA bispecific antibody is administered according to a regimen comprising at least one cycle of (i) administering the anti-DOTA bispecific antibody (so that the relevant tumor cells are optionally saturated), (ii) administering the bischelate of formula II and optionally the clearing agent, and (iii) optionally administering additional bischelate of formula II and / or the clearing agent without further administration of the anti-DOTA bispecific antibody. In some embodiments, the method may comprise multiple such cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cycles).

[0065] Additionally or alternatively, in some embodiments of the method, the tumor antigen targets are GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence) ), prostate cancer PSM, PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, lung resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le yThe antigen is selected from the group consisting of E-cadherin, V-cadherin, and EpCAM. In addition or alternatively, in some embodiments of the method, the anti-DOTA bispecific antibody and / or bischelate is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally.

[0066] In one embodiment, the present disclosure provides a method for enhancing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, comprising the steps of administering an effective amount of a complex comprising a bischelate of formula II and a bispecific antibody that recognizes and binds to the bischelate and a tumor antigen target, wherein the complex is configured to localize to a tumor expressing the tumor antigen target recognized by the bispecific antibody of the complex. The complex may be administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. In some embodiments, the subject is human.

[0067] In another embodiment, the present disclosure provides a method for treating cancer in a subject requiring it, comprising the steps of (a) administering to the subject an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target, and (b) administering to the subject an effective amount of a bischelate of formula II configured to bind to the anti-DOTA bispecific antibody. The anti-DOTA bispecific antibody is administered under conditions and for a sufficient period of time (e.g., according to a drug regimen) sufficient to saturate tumor cells. In some embodiments, unbound anti-DOTA bispecific antibody is removed from the bloodstream after administration of the anti-DOTA bispecific antibody. In some embodiments, the bischelate of formula II is administered after a period of time that may be sufficient to allow clearance of the unbound anti-DOTA bispecific antibody. In some embodiments, the subject is human.

[0068] Therefore, in some embodiments, the method further includes the step of administering an effective amount of a clearing agent to the target before administering the bischelate. The bischelate may be administered at any point between 1 minute and 4 days or more after administration of the anti-DOTA bispecific antibody. For example, in some embodiments, the bischelate is administered at 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 1.25 hours, 1.5 hours, 1.75 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours, 5.5 hours, 6 hours, 6.5 hours, 7 hours, 7.5 hours, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, or any range within that range. Alternatively, bischelate may be administered at any point at least 4 days after administration of the anti-DOTA bispecific antibody.

[0069] The clearing agent may be a 500kD aminodextran-DOTA conjugate (e.g., 500kD dextran-DOTA-Bn(Y), 500kD dextran-DOTA-Bn(Lu), or 500kD dextran-DOTA-Bn(In)). In some embodiments, the clearing agent and the bischelate of formula II are administered without further administration of an anti-DOTA bispecific antibody. For example, in some embodiments, the anti-DOTA bispecific antibody is administered according to a regimen comprising at least one cycle of (i) administering the anti-DOTA bispecific antibody (so that the relevant tumor cells are optionally saturated), (ii) administering the bischelate of formula II and optionally the clearing agent, and (iii) optionally administering additional bischelate of formula II and / or the clearing agent without further administration of the anti-DOTA bispecific antibody. In some embodiments, the method may comprise multiple such cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more cycles).

[0070] A method for treating cancer in a subject requiring such treatment is also provided herein, comprising the step of administering an effective amount of a conjugate comprising a bischelate of formula II and a bispecific antibody that recognizes and binds to the bischelate and a tumor antigen target, wherein the conjugate is configured to localize to a tumor expressing the tumor antigen target recognized by the bispecific antibody of the conjugate. The therapeutic efficacy of such a conjugate may be determined by calculating the area under the curve (AUC) tumor:AUC normal tissue ratio. In some embodiments, the complex has an AUC tumor:AUC normal tissue ratio of approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:1.

[0071] A method for treating cancer may further include the step of sequentially, separately, or simultaneously administering at least one chemotherapeutic agent selected from the group consisting of nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosourea, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxin, platinum coordination complexes, vinca alkaloids, substituted ureas, methylhydrazine derivatives, corticosteroids, hormone antagonists, endostatins, taxol, camptothecin, SN-38, doxorubicin, doxorubicin analogs, antimetabolites, alkylating agents, antimitotic agents, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, apoptosis promoters, methotrexate, and CPT-11. In some embodiments, cancer is selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. In some embodiments, the subject is human. A method for treating cancer disclosed herein may further include the step of administering (a) a bischelate of formula II, or (b) a conjugate comprising the bischelate of formula II and a bispecific antibody that recognizes and binds to the bischelate and a tumor antigen target, followed by monitoring tumor progression over time.

[0072] kit This technology provides a kit comprising components suitable for treating or diagnosing cancer in patients. In one embodiment, the kit comprises the compounds of this technology, at least one anti-DOTA BsAb, and instructions for use. The kit may further comprise a clearing agent (e.g., 500kDa aminodextran or 500kD dextran-DOTA-Bn(Y) conjugated to DOTA) and / or one or more radionuclides.

[0073] In some embodiments, at least one anti-DOTA BsAb is used to identify GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, and MUC-16. It binds to tumor antigen targets selected from the group consisting of MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence), prostate cancer psm, PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, lung resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), and Ki-67. In addition or alternatively, in some embodiments, at least one anti-DOTA BsAb is CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le yThe antibodies bind to tumor antigen targets selected from the group consisting of antigens, E-cadherin, V-cadherin, and EpCAM. At least one anti-DOTA BsAb may be provided in the form of a pre-filled syringe or auto-injector pen containing a sterile liquid formulation or sterile lyophilized preparation of the antibody (e.g., Kivitz et al., Clin. Ther. 28:1619-29 (2006)).

[0074] Additionally or alternatively, in some embodiments of the kit of this technology, one or more radionuclides are: 213 Bi, 211 At, 225 Ac, 152 Dy, 212 Bi, 223 Ra, 219 Rn, 215 Po, 211 Bi, 221 Fr, 217 At, and 255 Selected from Fm. In addition or alternatively, in certain embodiments, one or more radionuclides are: 86 Y, 90 Y, 89 Sr, 165 Dy, 186 Re, 188 Re, 177 Lu, 67 Cu, 111 In, 67 Ga, 51 Cr, 58 Co, 99m Tc, 103m Rh, 195m Pt, 119 Sb, 161 Ho, 189m Os, 192 Ir, 201 Tl, 203 Pb, 68 Ga, 227 Th, and 64 Selected from the group consisting of Cu. If the kit components are not formulated for oral administration, the kit may include a device capable of delivering the kit components by some other route. Examples of such devices include syringes or inhalation devices (for parenteral administration).

[0075] The kit components may be packaged together or separated into two or more containers. In some embodiments, the containers may be vials containing sterile lyophilized preparations of DOTA hapten and / or BsAb compositions suitable for reconstitution. The kit may also contain one or more buffers suitable for reconstitution and / or dilution of other reagents. Other containers that may be used include, but are not limited to, pouches, trays, boxes, and tubes. The kit components may be packaged within the containers and maintained in a sterile state. [Examples]

[0076] (Example 1) Materials and methods for preparing the composition of this technology General information: DOTA-Bn-isothiocyanate (p-SCN-Bn-DOTA) was purchased from Macrocyclics, Inc. (Plano, TX), and amine-PEG4-DOTA was purchased from CheMatech (Dijon, France). Optima® grade hydrochloric acid was purchased from Thermo Fisher Scientific (Waltham, MA). Chelex-100 resin, 200-400 mesh, was purchased from Bio-Rad Laboratories (Hercules, CA). PD-10 gel filtration size exclusion column (containing 8.3 mL of Sephadex® G-25 resin / column) was purchased from GE Healthcare Life Sciences (Pittsburgh, PA). All other reagents and synthetic grade chemicals were purchased from Sigma-Aldrich (St. Louis, MO) and used without further purification. All solvents used for HPLC analysis (HPLC grade) and compound purification were also purchased from Thermo Fisher Scientific (Waltham, MA). All buffers and solutions were prepared using ultrapure water (resistivity of 18 MΩ-cm).

[0077] All liquid chromatography-mass spectrometry (LC / MS) data were acquired using a Waters Autopure system (Milford, MA) including the following instruments: 2767 Sample Manager, 2545 Binary Gradient Module, System Fluidics Organizer, 2424 Evaporative Light Scattering Detector, 2998 Photodiode Array Detector, and 3100 Mass Detector. HPLC solvents (solvent A, 0.05% TFA in water; solvent B, 0.05% TFA in acetonitrile) were filtered before use. The analytical method involved 5–25% solvent B over 10 minutes at a flow rate of 1.2 mL / min. Analytical columns: Waters XBridge BEH300 (Milford, MA), C4, 3.5 μm, 4.6 × 50 mm and C18, 4 μm, 4.6 × 50 mm. Preparative method: 5–25% solvent B over 30 minutes at a flow rate of 20 mL / min. Preparative column: Waters XBridge Prep (Milford, MA) C18, 4 μm, Optimum Bed Density, 19 × 150 mm.

[0078] All NMR data were acquired at ambient temperature using either a Bruker AV500 or AV600 instrument (Bruker, Billerica, MA). The following abbreviations were used: singleline (s), broad singleline (bs), doubleline (d), tripleline (t), quadrupleline (q), pentetline (p), double-double-doubleline (dd), and multiline (m). All PET imaging experiments were performed using a Focus 120 MicroPET camera (Siemens, Knoxville, TN) with a dedicated small animal scanner.

[0079] p-SCN-Bn-DOTA·Lu 3+ Complex: [ka] To a 0.4M solution of LuCl3·6H2O (142 mg, 365 μmol) in 0.6 mL of NaOAc, p-SCN-Bn-DOTA·2.5HCl·2.5H2O (50 mg, 73 μmol) was added. The mixture was stirred overnight at room temperature (approximately 21°C). Purification was performed using a C-18 column with a gradient of 0-40% ACN / water to obtain 31.2 mg (60.5%) of the major isomer and 8 mg (15.2%) of the minor isomer.

[0080] Isolated major isomer (p-SCN-Bn-DOTA·Lu 3+ (Complex) 1 H NMR (D2O): 7.24 (d, 2 H), 7.19 (d, 2 H), 3.62-3.48 (m, 3 H), 3.42-3.18 (m, 8 H), 2.95-3.1 (m, 2 H), 2.37-2.82 (m, 11 H), 2.12 (d, 1 H).C 24 H 30 LuN5O8S[M+1] + Calculated MS = 724.13, measured: 724.18, negative mode: 722.11. HPLC, C-18, 5-50% gradient of acetonitrile in water containing 0.01% TFA. Peak R in an 8-minute run. f =4.35 minutes.

[0081] (Example 2) Synthesis of DOTA.Lu3+-PEG4-DFO Scheme 1 is the DOTA.Lu of this technology 3+ A synthetic route for providing PEG4-DFO is provided. Experimental details of the synthesis will be provided thereafter. Scheme 1 [ka]

[0082] DOTA-Lu 3+ -PEG4-NHBoc p-SCN-Bn-DOTA·Lu 3+The complex (major isomer of Example 1) (30 mg, 41.5 μmol) and Boc-NH-PEG4-NH2 (17 mg, 50.5 μmol) were added to DMF (0.8 mL), followed by the addition of Et3N (35 μL). The resulting mixture was stirred overnight at room temperature (approximately 21°C). The solvent was removed by vacuum evaporation, and then the mixture was dried under high vacuum. The resulting product was used directly in the following reaction.

[0083] Bn-DOTA.Lu 3+ -PEG4-NH2.TFA [ka] DOTA-Lu 3+ -PEG4-NHBoc was dissolved in a 4:1 (v / v) solution of DCM / TFA (0.8 mL), and the resulting colorless mixture was stirred at room temperature (approximately 21°C) for 40 minutes. The solvent was then removed by vacuum evaporation, and the residue was purified by HPLC using a 5-40% acetonitrile (containing 0.05% TFA) gradient in water (containing 0.05% TFA) on a C-18 reversed-phase column. Subsequently, the desired DOTA-Lu was obtained by lyophilization. 3+ -PEG4-NH2TFA salt (21 mg, 53%) was obtained as a white foamy substance.

[0084] Dota-Lu 3+ -PEG4-DFO [ka] At room temperature (approximately 21°C), Dota-Lu in DMF (0.8 mL) 3+ Solutions of -PEG(4)-NH2.TFA salt (21 mg, 21.9 μmol) and DFO-SCN (18 mg, 23.9 μmol) were treated with Et3N (15 μL), and stirred overnight at room temperature. Volatile substances were then removed under vacuum, and the residue was purified by reverse-phase HPLC using a 5-50% acetonitrile (containing 0.05% TFA) gradient in water (containing 0.05% TFA). After lyophilization of suitable fractions, DOTA-Lu was used. 3+ -PEG4-DFO (37.2 mg, 91%) was isolated as a white foamy substance.1 ¹H NMR, D2O: 7.23-7.17 (m, 8 H), 3.70-2.90 (m, 45 H), 2.81-2.30 (m, 19 H), 2.16 (d, 1 H), 2.02-2.05 (m, 3 H), 1.60-1.51 (m, 8 H), 1.44-1.39 (m, 4 H), 1.22-1.19 (m, 6 H). LCMS: Rf within an 8-minute run: 3.63 min. 67 H 106 LuN 15 O 20 MS[M+1] calculated for S3 + =1712.64, [M+1] 2+ =856.32. Actual measurement: 856.81. Negative mode: Calculation, [M-1] 2- = 855.31. Actual measurement: 855.37.

[0085] In particular, DOTA-Lu 3+ By using different isothiocyanates in similar reactions using PEG4-NH2TFA, other compounds and compositions of this technology can be obtained. For example, by using PCTA-isothiocyanate (shown in Scheme 3 below) or a salt thereof (e.g., Tris-HCl salt of PCTA-isothiocyanate) instead of DFO-SCN, DOTA.Lu of this technology shown in Scheme 3 can be obtained. 3+ -PEG4-PCTA is obtained. Scheme 3 [ka]

[0086] (Example 3) Synthesis of DOTA.Lu3+-PEG4-DOTA Scheme 2 is the DOTA.Lu of this technology 3+ This document provides a synthetic pathway for providing PEG4-DOTA. Experimental details of the synthesis will be provided later. Scheme 2 [ka]

[0087] DOTA-PEG4-NHBoc At room temperature (approximately 21°C), P-SCN-Bn-DOTA (30 mg, 54.4 μmol) and Boc-NH-PEG4-NH2 (18 mg, 53.5 μmol) were dissolved in anhydrous DMF (0.7 mL), and the resulting solution was treated with Et3N (36 μL). The mixture was stirred overnight at room temperature. The solvent was then removed by vacuum evaporation, and the residue was dried under high vacuum. This was submitted directly to the next step.

[0088] DOTA-PEG4-NH2.TFA DOTA-PEG4-NHBoc was dissolved in 4:1 (v / v) DCM / TFA (0.8 mL), and the resulting colorless mixture was stirred at room temperature for 40 minutes. The volatile substances were then evaporated to dryness, and the residue was purified by reverse-phase C-18 HPLC using a 5-40% acetonitrile (containing 0.05% TFA) gradient in water (containing 0.05% TFA). After lyophilization of the appropriate fractions, DOTA-PEG4-NH2.TFA (20 mg, 47%) was obtained.

[0089] DOTA.Lu 3+ -PEG4-DOTA [ka] At room temperature (approximately 21°C), DOTA-PEG4-NH2.TFA salt (20 mg, 25.4 μmol) and DOTA.Lu 3+ -SCN major isomer complex (15.3 mg, 21.1 μmol) was mixed in anhydrous DMF (0.8 mL) and then treated with Et3N (15 μL). The reaction mixture was stirred overnight at room temperature under an argon atmosphere. The solvent was then removed by vacuum evaporation, and the residue was purified by reverse-phase C-18 HPLC using a 5-50% acetonitrile (containing 0.05% TFA) gradient in water (containing 0.05% TFA). The product-containing fraction was lyophilized to obtain the desired DOTA-PEG4-DOTA.Lu 3+ The (19.6 mg, 61%) monocomplex was isolated as a white foamy substance. 1¹H NMR, D2O: 7.30–7.15 (m, 8 H), 3.75–2.90 (m, 58 H), 2.82–2.36 (m, 11 H), 2.18–2.14 (m, 1 H). The subsequent multiplex also includes some water peaks. LCMS: R for an 8-minute HPLC run f =4.51 minutes. C 58 H 87 LuN 12 O 20 MS calculated with respect to S2, [M+1] + =1511.51, [M+1] 2+ =755.75, actual measurement: 756.25. Negative mode, [M-1] 2- = 754.82, actual measurement: 754.75.

[0090] (Example 4) DOTA.Lu 3+ - Synthesis of PEG4-NODAGA DOTA.Lu 3+ -PEG4-NODAGA [ka] At room temperature (approximately 21°C), p-SCN-Bn-DOTA·Lu 3+ The major isomer complex (20 mg, 27.6 μmol) and NH2-PEG4-NODAGA (17 mg, 28.6 μmol) were dissolved in anhydrous DMF (0.8 mL) and treated with Et3N (20 μL). The resulting mixture was stirred overnight at room temperature. The solvent was then removed by vacuum evaporation, and the colorless residue was purified by reverse-phase C-18 HPLC using a 5-40% acetonitrile (containing 0.05% TFA) gradient in water (containing 0.05% TFA). After lyophilization of the appropriate fractions, DOTA.Lu 3+ -PEG4-NODAGA (15.1 mg, 41%) was obtained as a white foamy substance. 1H NMR, D2O: 7.15-7.25 (m, 4 H), 3.94-3.91 (m, 1 H), 3.89-3.51 (m, 26 H), 3.45-2.81 (m, 24 H), 2.5-2.35 (m, 12 H), 2.20-2.18 (m, 1 H), 2.07-2.03 (m, 1 H), 1.97-1.94 (m, 1 H). Two closely spaced isomers were observed in LC-MS at ratios of 18% and 82%. Within an 8-minute run, the minor was 3.02 minutes R. f It is a major, and the major is 3.08 minutes R f It belonged to them. C 49 H 77 LuN 10 O 19 MS = 1316.45 for S. [M+1] + =1317.46, [M+1] 2+ = 658.73. Actual measurement: 659.35.

[0091] Alternatively, DOTA-Lu 3+ -PEG4-NH2TFA is reacted with the NHS ester of NODAGA ("NODAGA-NHS", CAS number 1407166-70-4, shown in Scheme 4) and excess base in DMF. After the reaction is complete (e.g., as shown by HPLC), DOTA.Lu is purified by reverse-phase C-18 HPLC and lyophilized. 3+ -PEG4-NODAGA may be obtained. Scheme 4 [ka]

[0092] In particular, other compounds of this technology can be obtained by using protocols similar to any of the procedures described above. For example, HOPO-NHS (shown in Scheme 5) can be obtained using DOTA-Lu 3+-PEG4-NH2TFA is reacted with excess base in DMF, and after the reaction is complete (for example, as shown by HPLC), DOTA-Lu is purified by reverse-phase C-18 HPLC and lyophilized. 3+ -PEG4-HOPO may also be obtained (as shown in scheme 5). Scheme 5. HOPO-NHS and DOTA-Lu 3+ -PEG4-HOPO

[0093] [ka]

[0094] (Example 5) TCMC-PEG4- nat LuDOTABin synthesis Scheme 6 [ka] DOTA-Lu 3+ -PEG4-NHBoc: DOTA-Lu in DMF (0.8 mL) 3+ Et3N (29 μL) was added to SCN (25.0 mg, 34.6 μmol) and BocNH-PEG4-NH2 (13.9 mg, 41.3 μmol). The mixture was stirred at room temperature for 5 hours. The solvent was removed under reduced pressure. The residue was purified by preparative reverse-phase C-18 HPLC using a gradient of 20:80 MeCN:H2O to 40:60 MeCN:H2O (both containing 0.05% TFA) over 10 minutes, and the product was obtained after lyophilization (14.0 mg, 38%).

[0095] DOTA-Lu 3+ -PEG4-NH2: DOTA-Lu3+-PEG4-NHBoc (14.0 mg, 13.2 μmol) in TFA:DCM (4:1, V:V) was stirred at room temperature for 40 minutes, and then the solvent was removed under reduced pressure. The residue was dried under high vacuum (2 hours) and submitted directly to the next step without further purification.

[0096] DOTA-Lu 3+ -PEG4-TCMC The above residue was dissolved in DMF (0.8 mL), and then TCMC-DOTA (10 mg, 18.3 μmol) and Et3N (40 μL) were added to the mixture. The reaction mixture was stirred overnight at room temperature. Volatile substances were removed under reduced pressure, and the residue was purified by preparative C-18 reversed-phase HPLC using a gradient of 5:95 MeCN:H2O to 40:60 MeCN:H2O (both containing 0.05% TFA) over 10 minutes. The product was obtained after lyophilization (16.19 mg, 81%). 1 HNMR (500 MHz, D2O): δ = 7.25-7.18 (m, 8 H), 3.82-3.2 (m, 40 H), 3.10-2.95 (m, 2 H), 2.83-2.38 (m, 28 H).MS: Calculation: 1507.6[M+H] + Actual measurement: 1507.5.

[0097] (Example 6) Radiosynthesis of compounds using this technology Radiochemistry was performed in a properly shielded chemical fume hood equipped with an electron flow monitor and a sliding lead glass window. Radioactivity was measured using an ACRC-55tR dose calibrator with the manufacturer's recommended calibration settings (Capintec Inc., Forham Park, New Jersey). The buffers and water used for radiochemical synthesis were treated with 5% w / v Chelex ion exchange resin (BT Chelex 100 resin, Bio-Rad Inc., Hercules, California) to remove accidental heavy metals. Plastic products (pipette tips and microcentrifuge tubes) were trace metal grade / RNA grade. RadioHPLC was performed on a Shimadzu Prominence HPLC system consisting of a Flow-Count B-FC-1000 with an LC-20AB dual pump module, DGU-20A3R degasser, SIL-20ACHT autosampler, SPD-20AUV-Vis detector, and in-line PMT / NaI radioactivity detector. Separation was performed using analytical 4.6 × 250 mm Gemini-NXC18 or Fusion RP C18 HPLC columns (Phenomenex, Inc., Trans, California). Unless otherwise noted, HPLC conditions were: solvent A - 10 mM pH 5 NH4OAc, B - CH3CN, flow rate 1.0 mL / min, λ = 254 nm, injection volume 10–50 μL, gradient: 0%B–40%B, 10 minutes. Radioactive metal-free samples, reaction mixtures, and purified products were diluted 1:5 in 5 mM DTPA before analysis.

[0098] [ 203 Radiosynthesis of Pb]TCMC-PEG4-LuDOTA [ka] 15 μL of 0.5 M HCl (Lantheus Medical Imaging, Billerica, Massachusetts) 203PbCl2 (39.2 MBq / 1.06 mCi) was transferred to a 1.5 mL metal-free microcentrifuge tube and diluted with 200 μL of 0.5 M chelated aqueous solution of NH4OAc (pH 5.3), and gradually mixed. 10 μL of 1 mM TCMC-PEG4-LuDOTA (10 nmol) was added and gradually mixed, and the mixture was placed in a heat block set to 40°C. After 30 minutes, the reaction mixture was temporarily cooled, and then the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The column is slowly washed dropwise with 200 μL of water, the column is gradually purged with nitrogen gas, and then the product is slowly eluted dropwise with 200 μL of ethanol into a washed 2 mL microcentrifuge tube, diluted to 2.0 mL with normal physiological saline (Hospira, Lake Forest, Illinois), and filtered aseptically. [ 203 Pb]TCMC-PEG4-LuDOTABn(36.1MBq(975μCi), yield 92%, A M =3.9 MBq / nmol (10⁶ μCi / nmol) was obtained. Radio-HPLC confirmed that no free radioactive metals remained (98.1% radiochemical purity; major isomer t R =10.8 minutes).

[0099] [ 89 Radiosynthesis of Zr]DFO-PEG4-LuDOTA [ka] [ 50 μL of 1.0 M oxalic acid (Cyclotron Core Facility MSKCC) 89[Zr]Zr oxilate 2 (67.7 MBq / 1.83 mCi) was transferred to a 1.5 mL metal-free microcentrifuge tube, neutralized with approximately 45 μL of equimolar metal-free 1.0 M Na2CO3, and then diluted with 400 μL of metal-free 0.5 M HEPES buffer (pH 7.5) and mixed. DFO-PEG4-LuDOTA (9.2 nmol, 9.2 μL of 1.0 mM solution in water) was added and mixed, and the mixture was placed in a heat block at 40°C. After 60 minutes, the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The SPE cartridge was washed with 200 μL of water and slowly air-dried with nitrogen gas. The product was then slowly eluted with 200 μL of ethanol and dropped into a washed 2 mL microcentrifuge tube. The eluate was diluted to 2 mL with normal physiological saline (Hospira, Lake Forest, Illinois), and aseptically filtered. 89 44 MBq of [Zr]DFO-PEG4-LuDOTA was obtained (1.2 mCi; yield 66%, AM = 7.4 MBq (0.2 mCi) / nmol). Using this stock solution, doses for PET imaging and in vivo distribution were prepared (3.7 MBq / 100 μCi; 0.5 nmol). Radio-HPLC (solvent A: 0.1% TFA, B: CH3CN) of the crude and purified materials confirmed that no detectable free radioactive metals remained (major isomer t R (=10.7 minutes, 99%+ conversion).

[0100] [ 177 Radiosynthesis of Lu]DOTABn-PEG4-LuDOTA [ka] 0.05M HCl (NIDC / MURR; Missouri University Research Reactor, Columbia, Missouri) 19 μL in [ 177Lu]LuCl3 (38 MBq / 1.03 mCi) was transferred to a 1.5 mL metal-free microcentrifuge tube and diluted with 100 μL of metal-free 0.5 M NH4OAc (pH 5.3), and gradually mixed. DOTABn-PEG4-LuDOTABn (5 nmol, 5 μL of 1 mM solution in water) was added and gradually mixed, and the mixture was placed on an 80°C heat block for 60 minutes. After cooling for 5 minutes, the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The column was slowly washed dropwise with 200 μL of water and gradually air-dried with nitrogen gas. The product is slowly eluted with 200 μL of ethanol, added dropwise to a washed 2 mL microcentrifuge tube, diluted to 2.0 mL with normal physiological saline (Hospira, Lake Forest, Illinois), and filtered aseptically. [ 177 Lu]DOTABn-PEG4- nat LuDOTA was obtained (33.7 MBq (0.91 mCi), yield 88%, A M =7.4 MBq / nmol (0.2 mCi / nmol). Radio-HPLC analysis of the crude and purified materials confirmed that no free radioactive metals remained (99+% radiochemical purity; major isomer t). R =9.3 minutes).

[0101] [ 86 Radiosynthesis of Y]DOTABn-PEG4-LuDOTA [ka] 0.04M HCl (MDACC CRF; Cyclotron Radiochemistry Facility MD Anderson Cancer Center, Houston, Texas) in 5 μL [ 86YCl3 (4.7 MBq / 126 μCi) was transferred to a metal-free 0.5 mL microcentrifuge tube and diluted with 50 μL of metal-free 0.5 M NH4OAc (pH 5.3), and gradually mixed. DOTABn-PEG4-LuDOTABn (2 nmol, 2 μL of 1 mM solution in water) was added and gradually mixed, and the mixture was placed on an 80°C heat block for 60 minutes. After cooling for 5 minutes, the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The column was slowly washed dropwise with 200 μL of water and gradually air-dried with nitrogen gas. The product is slowly eluted with 200 μL of ethanol, added dropwise to a washed 2 mL microcentrifuge tube, diluted to 2.0 mL with normal physiological saline (Hospira, Lake Forest, Illinois), and filtered aseptically. [ 86 Y]DOTABn-PEG4- nat LuDOTA was obtained (1.38 MBq (37.2 μCi), yield 29%, A M =2.3 MBq / nmol (63 μCi / nmol). Radio-HPLC confirmed that no free radioactive metals remained (99+% radiochemical purity; major isomer t). R =9.15 minutes).

[0102] [ 68 Radiosynthesis of Ga]NODAGA-PEG4-LuDOTA [ka] GalliaPharm 68 Ge / 68 Ga generator (Eckert & Ziegler Radiopharma GmbH, Berlin, Germany) in 1 mL of 0.1 M HCl 68Ga]GaCl3 (175 MBq / 4.7 mCi) was eluted and transferred to a 2 mL metal-free microcentrifuge tube. The solution was diluted with 500 μL of 0.5 M NH4OAc chelate aqueous solution (pH 5.3) and gradually mixed. NODAGA-PEG4-LuDOTA (2 nmol in 20 μL of water) was added and gradually mixed. The tube was placed in an 80°C heat block for 15 minutes. After cooling for 5 minutes, the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The column was washed with 200 μL of water, air-dried with nitrogen gas, and the product was slowly eluted with 200 μL of ethanol and added dropwise to a washed 1.5 mL microcentrifuge tube. The volume of the eluate was reduced to approximately 50 μL under a flow of dry nitrogen gas, diluted with 2 mL of normal physiological saline (Hospira, Lake Forest, Illinois), and then sterile filtered. 68 Ga]NODAGA-PEG4-LuDOTA 141 MBq was obtained (3.8 mCi; yield 81%, A M =65 MBq / nmol (1.8 mCi / nmol). Using this stock solution, a dose for PET imaging was prepared (9.6 MBq / 260 μCi; 0.15 nmol), and further diluted with sterile saline for the in vivo distribution dose (6.5 MBq / 175 μCi; 0.1 nmol). RadioHPLC of the crude and purified materials confirmed that no free radioactive metals remained (major isomer t). R (=8.1 minutes, 99+% conversion).

[0103] [ 64 Radiosynthesis of Cu]NODAGA-PEG4-LuDOTA [ka] 4 μL (Washington University, St. Louis) of [ 64Cu]CuCl2 (38.1 MBq / 1.03 mCi) was transferred to a 1.5 mL metal-free microcentrifuge tube and diluted with 30 μL of 0.5 M NH4OAc chelate aqueous solution (pH 5.3), and gradually mixed. NODAGA-PEG4-LuDOTA (3 nmol) in 30 μL of buffer was then added and gradually mixed. After 5 minutes, the entire volume was gravity-loaded into a 30 mg Strata-X SPE cartridge (Phenomenex, Trans, California) equilibrated with 1 mL of ethanol and 1 mL of water. The reaction tube was rinsed with water (100 μL) and passed through to the cartridge. The column was slowly washed dropwise with 200 μL of water, slowly air-dried with nitrogen gas, and then the product was slowly eluted with 200 μL of ethanol and added dropwise to a washed 1.5 mL microcentrifuge tube. The volume of the eluate was reduced to approximately 50 μL by flowing dry nitrogen gas, diluted with normal physiological saline (Hospira, Lake Forest, Illinois), and aseptically filtered, [ 64 26.1 MBq (0.71 mCi; 68% yield) of Cu]NODAGA-PEG4-LuDOTA was obtained. Using this stock solution, a dose for PET imaging was prepared (11 MBq / 300 μCi; 1 nmol), and further diluted with sterile saline for in vivo distribution dose (1.9 MBq / 51 μCi; 0.15 nmol). RadioHPLC of the crude and purified materials confirmed that no free radioactive metals remained (99+% radiochemical purity; A). M (=12.7 MBq / nmol).

[0104] (Example 7) Stability of radionuclide-containing compounds in this technology In human serum [ 203 Pb] TCMC-PEG4-LuDOTA Stability: [ 203Pb]TCMC-PEG4-LuDOTA (88 μCi in 25 μL of PBS) was gradually mixed with 1 mL of human serum (Equitech-Bio) and incubated at 37°C. At three time points (1.5, 3, and 24 hours), 100 μL of the sample was taken and placed in a microcentrifuge tube. Each sample was treated with 200 μL of acetonitrile:methanol in a 3:1 ratio to precipitate the protein, and then centrifuged at 10,000 × g at 4°C for 10 minutes. The supernatant 200 μL was then removed, and the volume was reduced under a nitrogen gas stream for 20 minutes. The concentrate was diluted with 100 μL of 1 mL of MEDTA to release the protein. 203 Pb was chelated, and then 50 μL of each sample was analyzed by radio-HPLC. Independently prepared [ 203 Radio HPLC calibration using Pb]EDTA allows for [ 203 Regarding Pb]EDTA, a retention time of 2.2 minutes was observed. However, all three samples showed detectable levels of [ 203 Pb]EDTA could not be obtained, and therefore, when incubated in human serum at 37°C for 24 hours, [ 203 Decomposition of Pb]TCMC-PEG4-LuDOTA could not be verified.

[0105] [ 203 Plasma clearance of Pb]TCMC-PEG4-LuDOTA: In the tail veins of 5 female nude athymia mice (20-25g), [Pb]TCMC-PEG4-LuDOTA in 200 μL of sterile saline was administered. 203 Pb]PbTCMC-PEG4-LuDOTA95±2.4μCi was administered intravenously. At 5, 15, 30, 60, and 90 minutes after injection, the animals were euthanized by CO2 asphyxiation, and 0.5–1.0 mL of blood was immediately collected by intracardiac puncture and transferred to tubes containing EDTA anticoagulant on ice. The samples were centrifuged (10,000 × g, 4°C for 10 minutes). The radioactivity in 100 μL of plasma sample was counted using a PerkinElmer Wizard3 gamma counter with an energy window of 150–500 keV.

[0106] Raw data from a mouse plasma clearance study is provided below: [Table 1]

[0107] Using a calibration count for Pb-203 within a gamma counter window (150-500 keV), the radioactivity in each sample was calculated, corrected for decay over injection time, and then normalized by the injection dose in each animal. The percentage injection dose per gram (%ID / g) at each time point was calculated according to the following formula:

number

[0108] (Example 8) In vivo distribution properties of compounds of this technology Positron emission (PET) isotope gallium-68 68 DOTA-PRIT using Ga) was able to accelerate the development of companion PET diagnostics, 68 Antibody affinity for Ga-benzyl-DOTA was low (Orcutt KD, et al. (2012) Mol Cancer Ther, 11(6): 1365-72).

[0109] [ 89[Zr]DFO-PEG4-LuDOTA. Female thymic nude mice with scGPA33-expressing SW1222 xenografts were given HuA33-C825 (from Cheal, et al. Eur J Nucl Med Mol Imaging. 2016 May;43(5):925-937) 0.25 mg (1.19 nmol) at t=-28 hours, followed by 16-N-acetylgalactosamine-DOTA(Y) clearing agent; 25 μg (2.76 nmol) at t=-4 hours, and [ 89 Zr]DFO-PEG4-LuDOTA was administered to non-tumor controls at t=0. 89 Zr]DFO-PEG4-LuDOTA was administered. Mice that have undergone PRIT [ 89 The animals were killed 4 hours after injection of Zr]DFO-PEG4-LuDOTA, and at the same time [ 89 Intravitational distribution was evaluated by sacrificing animals that were given only Zr]DFO-PEG4-LuDOTA 4 hours after injection. Figure 5 shows [ 89 Representative PET maximum intensity projection images of two different mice that underwent PRIT using Zr]DFO-PEG4-LuDOTA are shown. The images are [ 89 Zr]DFO-PEG4-LuDOTA was obtained 4 hours after injection.

[0110] As shown in Figure 2, BsAb huA33-C825 and [ 89 In animals treated with PRIT using [Zr]DFO-PEG4-LuDOTA, blood, tumor, and renal uptake at 4 hours post-injection were 0.92±0.12%ID / g, 9.30±2.88%ID / g, and 6.45±0.85%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 10.1±2.0 and 1.4±0.3 for blood and kidney, respectively. 89 The blood uptake of Zr]DFO-PEG4-LuDOTA alone was 0.09±0.02 IA / g% 4 hours after injection, indicating very little uptake from normal tissue. See Figure 3. 89 The blood half-life of Zr]DFO-PEG4-LuDOTA was determined to be 11.88 minutes (R2 =0.9701). [ 89 The whole-body half-life of Zr]DFO-PEG4-LuDOTA was determined to be 59.76 minutes (R 2 (=0.8914). See Figure 4.

[0111] [ 68 Ga]NODAGA-PEG4-LuDOTA. DOTA.Lu 3+ -PEG4-NODAGA 68 We radiated labeling with Ga and characterized the radiostability, 68 Ga]NODAGA-PEG4-LuDOTA hapten (as specified herein) 68 In vitro and in vivo studies were conducted to determine whether pre-targeting of tumors with GPA33 (also known as "Ga-NODAGA-Proteus-DOTA") was feasible. Thymic nude mice carrying GPA33-expressing human colorectal cancer SW1222 xenografts were used as a model for anti-GPA33 benzyl-DOTA-PRIT.

[0112] DOTA.Lu 3+ -PEG4-NODAGA was synthesized from a non-radioactive lutetium-175 complex of amine-PEG4-NODAGA and 2-(4-isothiocyanatobenzyl)-DOTA. DOTA.Lu 3+ Radiolabeling of PEG4-NODAGA is typically performed using 2 nmol DOTA.Lu in 0.5 M sodium acetate pH 5.3. 3+ -PEG4-NODAGA contains approximately 185 MBq of eluted from the generator. 68 This was carried out by mixing Ga]GaCl3 and incubating at 80°C for 15 minutes (molar activity at the end of synthesis: 70 MBq / nmol; radiochemical yield: <98%; radiochemical purity: 98%).

[0113] Female thymic nude mice carrying scGPA33-expressing SW1222 xenografts were administered HuA33-C825 0.25 mg (1.19 nmol) at t=-28 hours (from Cheal, et al. Eur J Nucl Med Mol Imaging. 2016 May;43(5):925-937), followed by 16-N-acetylgalactosamine-DOTA(Y); 25 μg (2.76 nmol) at t=-4 hours, and at t=0 hours [ 68 Ga]DO3A-PEG4-LuDOTA or [[ 68 Ga]NODAGA-PEG4-LuDOTA was administered. Figure 7 shows [ 68 This image shows a typical PET image (coronal section) of a mouse that underwent PRIT using Ga]NODAGA-PEG4-LuDOTA. 68 The tumor was obtained 1 hour after injection of Ga]NODAGA-PEG4-LuDOTA. The tumor was clearly visible in the shoulder region.

[0114] As shown in Figure 6, BsAb huA33-C825 and [ 68 In animals treated with PRIT using Ga]NODAGA-PEG4-LuDOTA, blood, tumor, and renal uptake at 1 hour post-injection were 1.29±0.57%ID / g, 16.44±4.75%ID / g, and 1.23±0.25%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 12.7±3.9 and 13.4±2.7 for blood and kidney, respectively. In contrast, BsAb huA33-C825 and [ 68 Animals treated with PRIT using Ga]DO3A-PEG4-LuDOTA showed tumor-to-organ activity ratios of approximately 4.6±2.1 and 7.8±3.5 for blood and kidney, respectively. Therefore, the tumor-to-organ activity ratios for blood and kidney were [ 68 Compared to Ga]DO3A-PEG4-LuDOTA [ 68 In Ga]NODAGA-PEG4-LuDOTA, the levels were at least 1.7 to 2.7 times higher.

[0115] In an in vitro plasma stability study at 37°C using mouse serum, no significant demetallation occurred over 1 hour, and minimal serum-protein binding of radioactivity was observed. Female thymic nude mice with scGPA33-expressing SW1222 xenografts were administered HuA33-C825 0.25 mg (1.19 nmol) at t=-28 hours (from Cheal, et al. Eur J Nucl Med Mol Imaging. 2016 May;43(5):925-937), followed by 16-N-acetylgalactosamine-DOTA(Y); 25 μg (2.76 nmol) at t=-4 hours, and [ 68 Ga]NODAGA-PEG4-LuDOTA was administered. Regarding the calculation of moles, the prepared dose was [ 68 Regarding Ga]NODAGA-PEG4-LuDOTA, the amount was 132 μCi. 71 μCi [2.62 MBq] (75 pmol) was administered to mice. As shown in Figure 8, a series of in vivo distribution experiments were performed using pre-targeted [ 68 Rapid tumor targeting, combined with renal clearance, was demonstrated by administering [Ga]NODAGA-PEG4-LuDOTA (4 MBq, 67 pmol) via injection (pi) at 5, 15, 30, and 60 minutes. At 60 minutes via pi, tumor uptake was observed per gram per injection. 68 Ga-dose (%ID / g) reached approximately 10 percent of the dose, with minimal accumulation in normal tissues, including blood and kidneys (both approximately 1%ID / g). Maximum tumor uptake (8-10 IA / g%) was achieved within 15 minutes post-injection, and maximum tumor-to-blood and tumor-to-kidney ratios (both approximately 10:1) were achieved within 30 minutes post-injection. See Figure 9.

[0116] [ 64 Cu]NODAGA-PEG4-LuDOTA. Female thymic nude mice with scGPA33-expressing SW1222 xenografts were administered HuA33-C825BsAb 0.25 mg (1.19 nmol) at t=-28 hours, followed by 16-N-acetylgalactosamine-DOTA(Y); 25 μg (2.76 nmol) at t=-4 hours, and [ 64Cu]NODAGA-PEG4-LuDOTA was administered. Figure 11 shows [ 64 This image shows a typical PET image (coronal section) of a mouse that underwent PRIT using Cu]NODAGA-PEG4-LuDOTA. 64 The tumor was obtained approximately 24 hours after injection of Cu]NODAGA-PEG4-LuDOTA 300μ Curie. The tumor is clearly visible in the shoulder region ("T").

[0117] As shown in Figure 10, BsAb huA33-C825 and [ 64 In animals treated with PRIT using Cu]NODAGA-PEG4-LuDOTA, blood, tumor, and renal uptake at 24 hours post-injection were 0.22±0.03%ID / g, 3.53±0.55%ID / g, and 0.41±0.03%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 15.8±1.7 and 8.6±0.7 for blood and kidney, respectively.

[0118] [ 177 Lu]DOTABn-PEG4-LuDOTA. SW1222 tumor-bearing mice (n=4~6) were given huA33-C825 250 μg, followed 24 hours later by dentrimer-clearing agent (10% (w / w), 25 μg), and then 4 hours later [ 177 Lu]Lu-aminobenzyl DOTA (as shown in Scheme 7 below) or [ 177 Lu]DOTABn-PEG4-LuDOTA (in this specification, "[ 177 [Lu]Lu-GeminiDOTA (also known as Lu]Lu-GeminiDOTA) was administered (see Figure 12A for administered moles / activity). As shown in Figures 12A-12B, BsAb huA33-C825 and [ 177 In animals treated with PRIT using Lu]DOTABn-PEG4-LuDOTA, blood, tumor, and kidney uptake at 24 hours post-injection were 0.14±0.02%ID / g, 5.07±0.38%ID / g, and 0.48±0.05%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 36.2±5.8 and 10.6±1.3 for blood and kidney, respectively. Furthermore, in tumors, [ 177Long-term retention of Lu]DOTABn-PEG4-LuDOTA was observed (both from this data and other data), particularly [ 177 This is a significant advantage in that it delivers a much higher dose to solid tumors than Lu]DOTABn-PEG4-LuDOTA. Scheme 7.[ 177 Lu]Lu-aminobenzyl DOTA [ka]

[0119] [ 203 Pb]TCMC-PEG4-LuDOTA and [ 203 Pb]DO3A-PEG4-LuDOTA. SW1222 tumor-bearing mice (n=4) were given huA33-C825 250μg, followed 24 hours later by dentrimer-clearing agent (10% (w / w), 25μg), and then 4 hours later [ 203 Pb]TCMC-PEG4-LuDOTA (in this specification, "[ 203 Pb]TCMC-Proteus-DOTA (also known as [ 203 Pb]DO3A-PEG4-LuDOTA (as specified herein) 203 [Pb]Proteus-DOTA (also known as "Proteus-DOTA") was administered (see Figure 13 for the moles administered / activity). 203 The structure of Pb]DO3A-PEG4-LuDOTA is shown in Scheme 8 below. Further [ 203 For information regarding Pb]DO3A-PEG4-LuDOTA, please refer to International Application PCT / US2018 / 040911, filed on July 5, 2018, and WO2019 / 010299, published on January 10, 2019. Scheme 8.[ 203 Pb]DO3A-PEG4-LuDOTA(「[ 203 (Also known as Pb[Proteus-DOTA])

[0120] [ka]

[0121] As shown in Figure 13, BsAb huA33-C825 and [ 203 In animals treated with PRIT using [Pb]TCMC-Proteus-DOTA, blood, tumor, and renal uptake at 24 hours post-injection were 0.31±0.12%ID / g, 27.79±7.38%ID / g, and 1.49±0.07%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 89.6±20.6 and 18.6±2.5 for blood and kidney, respectively.

[0122] As in another comparison, a group of SW1222 tumor-bearing mice (n=4) were given huA33-C825 250 μg, followed 24 hours later by dentrimer-clearing agent (10% (w / w), 25 μg), and then 4 hours later by "[ 111 In] Proteus-DOTA(Lu)" or "[ 111 One of the following [Proteus-DOTA(Gd)] (shown in Scheme 9 below) was administered. Further [ 111 In] Proteus-DOTA(Lu) and [ 111 For information regarding Proteus-DOTA(Gd), please refer to International Application PCT / US2018 / 040911, filed on July 5, 2018, and WO2019 / 010299, published on January 10, 2019. Scheme 9.[ 111 In] Proteus-DOTA(Lu) or [ 111 In] Proteus-DOTA(Gd)

[0123] [ka]

[0124] As shown in Figure 14, BsAb huA33-C825 and [ 111In animals treated with PRIT using [In]Proteus-DOTA(Lu), blood, tumor, and renal uptake at 24 hours post-injection were 0.63±0.31%ID / g, 9.25±2.72%ID / g, and 0.67±0.13%ID / g, respectively, corresponding to tumor-to-organ activity ratios of approximately 14.6±4.2 and 13.9±2.5 for blood and kidney, respectively. Figure 14 shows BsAb huA33-C825 and [ 111 In animals treated with PRIT using [In]Proteus-DOTA(Gd), blood, tumor, and renal uptake at 24 hours post-injection were 0.46±0.21%ID / g, 7.66±4.74%ID / g, and 0.58±0.11%ID / g, respectively, further demonstrating that these corresponded to tumor-to-organ activity ratios of approximately 16.6±6.3 and 13.3±4.3 for blood and kidney, respectively. These results demonstrate that the composition of this technology is useful for in vivo diagnostic imaging and pre-targeted radioimmunotherapy.

[0125] Equal portions The Art should not be limited by the specific embodiments described herein, which are intended merely as examples of individual aspects of the Art. Many modifications and variations of the Art can be made without departing from the spirit and scope of the Art, as will be obvious to those skilled in the art. In addition to the methods and apparatus described herein, functionally equivalent methods and apparatus within the scope of the Art will be obvious to those skilled in the art from the foregoing description. Such modifications and variations are intended to fall within the scope of the Art. It should be understood that the Art is not limited to specific methods, reagents, compounds, compositions, or biological systems, which are, of course, subject to change. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments and is not intended to limit them. In addition, if any feature or aspect of the present disclosure is described by a Markush group, it will be recognized by those skilled in the art that the present disclosure is also described by any individual component or subgroup of components of any of the Markush groups.

[0126] As will be understood by those skilled in the art, for all purposes, and in particular with respect to providing descriptive explanations, all scopes disclosed herein also encompass all possible sub-scopes and combinations thereof. Any enumerated scope can be readily recognized as sufficiently descriptive and enabling that the scope may be divided into at least equal 1 / 2, 1 / 3, 1 / 4, 1 / 5, 1 / 10, etc. As a non-limiting example, each scope discussed herein can be readily divided into a lower third, a middle third, and an upper third, etc. Also as will be understood by those skilled in the art, all phrases such as “at most,” “at least,” “greater than,” and “less than” include the number described and refer to a scope that can subsequently be divided into the sub-scopes discussed above. Finally, as will be understood by those skilled in the art, a scope includes each individual component. Thus, for example, a group having 1 to 3 cells refers to a group having 1, 2, or 3 cells. Similarly, a group having 1 to 5 cells refers to a group having 1, 2, 3, 4, or 5 cells, and so on. All patents, patent applications, provisional applications, and publications mentioned or cited herein, including all figures and tables, are incorporated by reference in their entirety, to the extent that they do not conflict with the express teachings herein.

[0127] This technology may include, but is not limited to, the features and combinations of features listed in the following paragraphs, and it should be understood that the following sections should not be construed as limiting the scope of the claims appended to this specification, nor should they obligate all such features to be included in such claims: A. Compounds of formula I [ka] (I) or a pharmaceutically acceptable salt thereof. (In the formula, M 1 Chelate 175 Lu 3+ , 45 Sc 3+ , 69Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ , 151 EU 3+ , 153 EU 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ , or 160 Gd 3+ and; R 1 teeth,

[0128] [ka] [ka] [ka] [ka] and; X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , and X 36 Each of these is independently either a lone pair of electrons (i.e., giving the oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 Each is independently either S or O; Q 1 is either S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22. B. Bischelates containing the compounds and radionuclide cations described in Section A. C. Bischelate is the one of formula II, as described in section B.

[0129] [ka] (II) or a pharmaceutically acceptable salt thereof. (In the formula, M 1 Chelate 175 Lu 3+ ,45 Sc 3+ , 69 Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ , 151 EU 3+ , 153 EU 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ , or 160 Gd 3+ and; R 2 teeth, [ka] [ka] [ka] [ka] and; M 2 Each instance is independent of R 2 It is a radioactive nuclide cation chelated by a group; X 1 , X 2 , X 3 , X 4, X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , and X 36 are each independently a lone pair (i.e., giving an oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 are each independently S or O; Q 1 is S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22) D.M 2 is the bischelating agent according to item C, which is an alpha particle emitting isotope, a beta particle emitting isotope, an Auger emitter, or a combination of two or more of them. E.M 2 is 213 Bi, 211 At, 225 Ac, 152 Dy,212 Bi, 223 Ra, 219 Rn, 215 Po, 211 Bi, 221 Fr, 217 At, or 255 A bischelate described in item C or D, which is Fm. FM 2 but, 86 Y, 90 Y, 89 Sr, 165 Dy, 186 Re, 188 Re, 177 Lu, or 67 A bischelate described in item C or D, which is Cu. GM 2 but, 111 In, 67 Ga, 51 Cr, 58 Co, 99m Tc, 103m Rh, 195m Pt, 119 Sb, 161 Ho, 189m Os, 192 Ir, 201 Tl, or 203 Bischelate, which is Pb, as described in item C or D. HM 2 but, 89 Zr, 68 Ga, 212 Pb, 227 Th, or 64 A bischelate described in item C or D, which is Cu. I. A complex comprising the compound described in Section A and a bispecific antibody that recognizes and binds to the compound and a tumor antigen target. J. A complex comprising a bischelate described in any one of sections B to H, and a bispecific antibody that binds to the bischelate and a tumor antigen target. K. Tumor antigen targets include GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence), prostate cancer psm, P RAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, pulmonary resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le y A complex as described in item I or J, selected from the group consisting of antigen, E-cadherin, V-cadherin, and EpCAM. L. Bispecific antibodies are converted to bischelate in the following concentrations: 100nM~95nM, 95~90nM, 90~85nM, 85~80nM, 80~75nM, 75~70nM, 70~65nM, 65~60nM, 60~55nM, 55~50nM, 50~45nM, 45~40nM, 40~35nM, 35~30nM, 30~25nM, 25~20nM, 20~15nM, 15~10nM, 10~5nM, 5~1nM, 1nM~950pM, 950pM~900pM, 900pM~850pM, 850pM~800pM, 800pM~750pM, and 750pM~700pM. , 700pM~650pM, 650pM~600pM, 600pM~550pM, 550pM~500pM, 500pM~450pM, 4 50pM~400pM, 400pM~350pM, 350pM~300pM, 300pM~250pM, 250pM~200pM, 200 K levels of 1 pM to 150 pM, 150 pM to 100 pM, 100 pM to 50 pM, 50 pM to 40 pM, 40 pM to 30 pM, 30 pM to 20 pM, 20 pM to 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2.5 pM, 2 pM, 1.5 pM, or less than 1 pM. d The complex described in item J or K, which is joined by [a specific link / method]. M. A method for detecting tumors in subjects requiring it, (a) A step of administering an effective amount of the complex described in any one of sections J to K, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody; (b) A step of detecting the presence of a tumor in the subject by detecting a level of radioactivity released by the complex that is higher than the standard value. A method that includes this. N. A method for selecting subjects for pre-targeted radioimmunotherapy, (a) A step of administering an effective amount of the complex described in any one of sections J to K, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody; (b) A step of detecting the level of radioactivity released by the complex, (c) If the level of radioactivity released by the complex is higher than the standard value, the process of selecting a target for pre-targeted radioimmunotherapy. A method that includes this. The method according to item M or N, wherein the level of radioactivity emitted by the complex is detected using positron emission tomography or single-photon emission computed tomography. P. The method described in any one of sections M to O, for subjects who have been diagnosed with cancer or are suspected of having cancer. Q. The method according to item P, wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. R. The method according to paragraph Q, wherein the brain cancer is a pituitary adenoma, meningioma, neuroblastoma, or craniopharyngioma. The method according to any one of items M to R, wherein the complex is administered into the cerebrospinal fluid or blood of the subject. The method according to any one of items M to S, wherein the T complex is administered intravenously, intramuscularly, intraarterially, into the subarachnoid space, intrajoint capsule, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. The method according to any one of sections M to T, wherein the level of radioactivity released by the complex is detected 4 to 24 hours after the complex is administered. V. The method according to any one of items M to U, wherein the radioactivity level released by the complex is expressed as a percentage of the injection dose per gram of tissue (%ID / g). W. The method according to any one of items M to V, wherein the ratio of radioactivity levels between tumor tissue and normal tissue is approximately 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:1. X. A method for increasing the sensitivity of tumors to radiation therapy in subjects diagnosed with cancer, (a) A step of administering an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target, (b) A step of administering an effective amount of the bischelate described in any one of sections B to H, configured to bind to an anti-DOTA bispecific antibody. A method that includes this. The method according to item X, further comprising the step of administering an effective amount of a clearing agent to the target before administering Y. bischelate. The method according to item Y, wherein the clearing agent is 500kD aminodextran-DOTA conjugate. AA. Tumor antigen targets include GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence), prostate cancer psm. PRAME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, pulmonary resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le y The method described in any one of items X to Z, selected from the group consisting of antigen, E-cadherin, V-cadherin, and EpCAM. AB. The method according to any one of items X to AA, wherein an anti-DOTA bispecific antibody is administered intravenously, intramuscularly, intraarterially, into the subarachnoid space, into the joint capsule, into the orbit, intradermally, intraperitoneally, transtracheally, subcutaneously, into the ventricle, orally, or intranasally. The method according to any one of paragraphs X to AB, wherein AC. bischelate is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. AD. A method for increasing the sensitivity of tumors to radiation therapy in subjects diagnosed with cancer, A step of administering an effective amount of the complex described in any one of sections J to L, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody. A method that includes this. The method according to item AD, wherein the AE complex is administered intravenously, intramuscularly, intraarterially, into the subarachnoid space, intrajoint capsule, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or intranasally. AF. A method for treating cancer in a subject that requires it, (a) A step of administering an effective amount of an anti-DOTA bispecific antibody configured to localize to a tumor expressing a tumor antigen target, (b) A step of administering an effective amount of the bischelate described in any one of sections B to H, configured to bind to an anti-DOTA bispecific antibody. A method that includes this. The method according to item AF, further comprising the step of administering an effective amount of a clearing agent to the target before administering AG. bischelate. AH. A method for treating cancer in a subject that requires it, A step of administering an effective amount of the complex described in any one of sections J to L, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody. A method that includes this. AI. Nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosourea, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxin, platinum coordination complexes, vinca alkaloids, urea substitutions, methylhydrazine derivatives, adrenal cortical depressants, hormone antagonists, endostatins, taxol, camptothecin, SN-38, doxo The method according to any one of sections AF to AH, further comprising the step of sequentially, individually, or simultaneously administering at least one chemotherapeutic agent selected from the group consisting of rubicin, doxorubicin analogs, anti-antimetabolites, alkylating agents, antimitotic agents, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, apoptosis promoters, methotrexate, and CPT-11. AJ. The method according to any one of items X to AI, wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer. A kit comprising the compound described in Section A of AK, at least one anti-DOTA BsAb, and instructions for use. A kit comprising a bischelate described in any one of sections B to H of AL, at least one anti-DOTA BsAb, and instructions for use. The kit according to section AK or AL, further comprising an AM clearing agent and / or one or more radionuclides. The kit described in Section AM, wherein the clearing agent is 500kD aminodextran-DOTA conjugate.

[0130] Other embodiments are shown in the following claims.

Claims

1. Compound of formula I 【Chemistry 1】 (I) or a pharmaceutically acceptable salt thereof. (In the formula, M 1 is chelation 175 Lu 3+ , 45 Sc 3+ , 69 Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ , 151 Eu 3+ , 153 Eu 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ or 160 Gd 3+ ; R 1 teeth, 【Chemistry 2-1】 【Chemistry 2-2】 [Chemistry 2-3] 【Chemistry 2-4】 And; X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , and X 36 Each of these is independently either a lone pair of electrons (i.e., giving an oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 Each is independently either S or O; Q 1 is S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22.

2. A bischelate comprising the compound described in claim 1 and a radionuclide cation, Bischelate is Equation II 【Transformation 3】 (II) It may be the same as or a pharmaceutically acceptable salt thereof, in the formula, M 1 is chelation 175 Lu 3+ , 45 Sc 3+ , 69 Ga 3+ , 71 Ga 3+ , 89 Y 3+ , 113 In 3+ , 115 In 3+ , 139 La 3+ , 136 Ce 3+ , 138 Ce 3+ , 140 Ce 3+ , 142 Ce 3+ , 151 Eu 3+ , 153 Eu 3+ , 159 Tb 3+ , 154 Gd 3+ , 155 Gd 3+ , 156 Gd 3+ , 157 Gd 3+ , 158 Gd 3+ or 160 Gd 3+ ; R 2 teeth, 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】 【Chemistry 4-4】 And; M 2 Each of these appears independently, R 2 It is a radioactive nuclide cation chelated by a group; X 1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , X 8 , X 9 , X 10 , X 11 , X 12 , X 13 , X 14 , X 15 , X 16 , X 17 , X 18 , X 19 , X 20 , X 21 , X 22 , X 23 , X 24 , X 25 , X 26 , X 27 , X 28 , X 29 , X 30 , X 31 , X 32 , X 33 , X 34 , X 35 , and X 36 Each of these is independently either a lone pair of electrons (i.e., giving an oxygen anion) or H; Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , and Y 9 Each is independently either S or O; Q 1 is either S or O; n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22, where, M 2 However, this may be an alpha particle-emitting isotope, a beta particle-emitting isotope, an Auger emitter, or a combination of two or more of these; or M 2 but, 213 Bi, 211 At, 225 Ac, 152 Dy, 212 Bi, 223 Ra, 219 Rn, 215 Po, 211 Bi, 221 Fr, 217 At, or 255 It may also be Fm; or M 2 but, 86 Y, 90 Y, 89 Sr, 165 Dy, 186 Re, 188 Re, 177 Lu, or 67 It may also be Cu; or M 2 but, 111 In, 67 Ga, 51 Cr, 58 Co, 99m Tc, 103m Rh, 195m Pt, 119 Sb, 161 Ho, 189m OS, 192 Ir, 201 Tl, or 203 It may be Pb; or M 2 but, 89 Zr, 68 Ga, 212 Pb, 227 Th, or 64 A bischelate that may be made of Cu.

3. A complex comprising the compound described in claim 1 and a bispecific antibody that recognizes and binds to the compound and a tumor antigen target.

4. A complex comprising a bischelate according to claim 2 and a bispecific antibody that binds to the bischelate and a tumor antigen target, The tumor antigen targets are GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence), prostate cancer psm, PRA ME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, pulmonary resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen-p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le y ) may be selected from the group consisting of antigen, E-cadherin, V-cadherin, and EpCAM; or Bispecific antibodies are available in the following concentrations: 100 nM–95 nM, 95–90 nM, 90–85 nM, 85–80 nM, 80–75 nM, 75–70 nM, 70–65 nM, 65–60 nM, 60–55 nM, 55–50 nM, 50–45 nM, 45–40 nM, 40–35 nM, 35–30 nM, 30–25 nM, 25–20 nM, 20–15 nM, 15–10 nM, 10–5 nM, 5–1 nM, 1 nM–950 pM, 950 pM–900 pM, 900 pM–850 pM, 850 pM–800 pM, 800 pM–750 pM, 750 pM–700 pM, and 700 pM. M to 650 pM, 650 pM to 600 pM, 600 pM to 550 pM, 550 pM to 500 pM, 500 pM to 450 pM, 450 pM to 400 pM, 400 pM to 350 pM, 350 pM to 300 pM, 300 pM to 250 pM, 250 pM to 200 pM, 200 pM to 150 pM, 150 pM to 100 pM, 100 pM to 50 pM, 50 pM to 40 pM, 40 pM to 30 pM, 30 pM to 20 pM, 20 pM to 10 pM, 9 pM, 8 pM, 7 pM, 6 pM, 5 pM, 4 pM, 3 pM, 2.5 pM, 2 pM, 1.5 pM, or K less than or equal to 1 pM. d A complex that may be bound to a bischelate.

5. A composition for use in a method for detecting tumors in an object requiring the use of the complex described in Claim 4, wherein the method is: (a) A step of administering an effective amount of the complex to a target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody; (b) A step of detecting the presence of a tumor in the subject by detecting the level of radioactivity released by the complex that is higher than the standard value. A composition containing the following:

6. A composition for use in a method for selecting a target for pre-targeted radioimmunotherapy, comprising the complex described in Claim 4, wherein the method is: (a) A step of administering an effective amount of the complex to a target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody; (b) A step of detecting the level of radioactivity released by the complex; (c) If the level of radioactivity released by the complex is higher than the standard value, the step of selecting a target for pre-targeted radioimmunotherapy. A composition containing the following:

7. The level of radioactivity emitted by the complex is detected using positron emission tomography or single-photon emission computed tomography; or The subject is diagnosed with cancer or suspected of having cancer, and the cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer; or The complex may be administered into the cerebrospinal fluid or blood of the subject; or The complex may be administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally or nasally; or The level of radioactivity released by the complex may be detected between 4 and 24 hours after the complex is administered; or The level of radioactivity released by the complex may be expressed as a percentage of the injection dose per gram of tissue (%ID / g); or The composition according to claim 5 or claim 6, wherein the ratio of radioactivity levels between tumor tissue and normal tissue may be 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, or 100:

1.

8. The composition according to claim 7, wherein the brain cancer is a pituitary adenoma, meningioma, neuroblastoma, or craniopharyngioma.

9. A composition comprising the bischelate described in Claim 2, for use in a method for increasing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, wherein the method is: (a) A step of administering an effective amount of an anti-DOTA bispecific antibody to a target, wherein the anti-DOTA bispecific antibody is configured to localize to a tumor expressing a tumor antigen target; (b) A step of administering an effective amount of the bischelate to a target, wherein the bischelate is configured to bind to an anti-DOTA bispecific antibody. Includes, A composition in which cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer.

10. The method further comprises the step of administering an effective amount of a clearing agent to the target before administering the bischelate; or The tumor antigen targets are GPA33, HER2 / neu, GD2, MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, MUM-1, CDK4, N-acetylglucosaminyltransferase, p15, gp75, beta-catenin, ErbB2, cancer antigen 125 (CA-125), carcinoembryonic antigen (CEA), RAGE, MART (melanoma antigen), MUC-1, MUC-2, MUC-3, MUC-4, MUC-5ac, MUC-16, MUC-17, tyrosinase, Pmel17 (gp100), GnT-V intron V sequence (N-acetylglucosaminyltransferase V intron V sequence), prostate cancer psm, PRA ME (melanoma antigen), β-catenin, EBNA (Epstein-Barr virus nuclear antigen) 1-6, p53, pulmonary resistance protein (LRP) Bcl-2, prostate-specific antigen (PSA), Ki-67, CEACAM6, colon-specific antigen-p (CSAp), HLA-DR, CD40, CD74, CD138, EGFR, EGP-1, EGP-2, VEGF, PlGF, insulin-like growth factor (ILGF), tenascin, platelet-derived growth factor, IL-6, CD20, CD19, PSMA, CD33, CD123, MET, DLL4, Ang-2, HER3, IGF-1R, CD30, TAG-72, SPEAP, CD45, L1-CAM, Lewis Y (Le y ) Selected from the group consisting of antigen, E-cadherin, V-cadherin, and EpCAM; or The composition according to claim 9, wherein the anti-DOTA bispecific antibody or bischelate is administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally, or nasally.

11. The composition according to claim 10, wherein the clearing agent is 500 kD aminodextran-DOTA conjugate.

12. A composition for use in a method for increasing the sensitivity of a tumor to radiotherapy in a subject diagnosed with cancer, comprising the complex described in Claim 4, wherein the method is: A step of administering an effective amount of the complex, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the complex's bispecific antibody; The complex may be administered intravenously, intramuscularly, intraarterially, subarachnoidally, intracapsularly, intraorbitally, intradermally, intraperitoneally, transtracheally, subcutaneously, intraventricularly, orally or nasally; or A composition in which cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer.

13. A composition for use in a method for treating cancer in a subject requiring the use thereof, comprising the bischelate described in Claim 2, wherein the method is: (a) A step of administering an effective amount of an anti-DOTA bispecific antibody to a target, wherein the anti-DOTA bispecific antibody is configured to localize to a tumor expressing a tumor antigen target; (b) A step of administering an effective amount of the bischelate to a target, wherein the bischelate is configured to bind to an anti-DOTA bispecific antibody. Includes, The method may further include a step of administering an effective amount of the clearing agent to the target before administering the bischelate; or A composition in which cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, stomach cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer.

14. A composition for use in a method for treating cancer in a subject requiring the use thereof, comprising the complex described in Claim 4, wherein the method is: A composition comprising the step of administering an effective amount of the complex to a target, wherein the complex is configured to localize to a tumor expressing a tumor antigen target recognized by the bispecific antibody of the complex, wherein the cancer may be selected from the group consisting of breast cancer, colorectal cancer, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, hepatocellular carcinoma, brain cancer, lung cancer, gastric cancer, pancreatic cancer, thyroid cancer, kidney or renal cancer, prostate cancer, melanoma, sarcoma, carcinoma, Wilms' tumor, endometrial cancer, glioblastoma, squamous cell carcinoma, astrocytoma, salivary gland cancer, vulvar cancer, penile carcinoma, leukemia, lymphoma, and head and neck cancer.

15. The aforementioned method involves nitrogen mustard, ethyleneimine derivatives, alkyl sulfonates, nitrosourea, gemcitabine, triazenes, folic acid analogs, anthracyclines, taxanes, COX-2 inhibitors, pyrimidine analogs, purine analogs, antibiotics, enzyme inhibitors, epipodophyllotoxin, platinum coordination complexes, vinca alkaloids, substituted ureas, methylhydrazine derivatives, adrenal cortical inhibitors, hormone antagonists, endostatins, taxol, camptothecin, and SN-38. The composition according to claim 13 or claim 14, further comprising the step of sequentially, separately, or simultaneously administering at least one chemotherapeutic agent selected from the group consisting of doxorubicin, doxorubicin analogs, antimetabolites, alkylating agents, antimitotic agents, anti-angiogenic agents, tyrosine kinase inhibitors, mTOR inhibitors, heat shock protein (HSP90) inhibitors, proteosome inhibitors, HDAC inhibitors, apoptosis promoters, methotrexate, and CPT-11.

16. A kit comprising the compound described in claim 1, at least one anti-DOTA BsAb, and instructions for use, which may further comprise a clearing agent and / or one or more radionuclides.

17. A kit comprising the bischelate described in claim 2, at least one anti-DOTA BsAb, and instructions for use, which may further comprise a clearing agent and / or one or more radionuclides.

18. The kit according to claim 16 or claim 17, wherein the clearing agent is 500 kD aminodextran-DOTA conjugate.

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