Radioimmunoconjugate and checkpoint inhibitor combination therapy

Abstract

Combination therapy comprising administering a radioimmunoconjugate and one or more checkpoint inhibitors. The present disclosure encompasses the insight that combining the inhibition of checkpoint proteins with a treatment that targets damage to cancer cells may provide a less toxic treatment with improved efficacy. Radioactive decay can cause direct physical damage (such as single- or double-stranded DNA breaks) or indirect damage (such as bystander or crossfire effects) to the biomolecules that make up cells. The present disclosure combines radioimmunoconjugates that target cancer cells with checkpoint inhibition to induce or improve immune responses against tumors. In some embodiments, the disclosed combination therapy reverses or treats cancer.

Description

[Technical field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 209,736, filed June 11, 2021, the entire contents of which are incorporated herein by reference for all purposes.

[0002] Sequence Listing This specification references a Sequence Listing (submitted electronically as a txt file entitled "FPI_021_Sequence_Listing.txt" on Jun. 10, 2022). The txt file was generated on Jun. 9, 2022 and is 19.2 kilobytes in size. The entire contents of the Sequence Listing are incorporated herein by reference. [Background technology]

[0003] background Cancer cells use multiple mechanisms to evade immune surveillance, including suppression of T cell activation.

[0004] The mammalian immune system relies on checkpoint molecules to distinguish between normal and foreign cells. Checkpoint molecules expressed on specific immune cells must be activated or inactivated to mount an immune response. Inhibiting checkpoint proteins promotes activation of the immune system.

[0005] Checkpoint inhibition is being explored as a method of cancer immunotherapy. By inhibiting checkpoint proteins, T cells can be activated and directed to attack cancer cells. However, checkpoint inhibition can cause the immune system to attack some normal cells in the body, which can lead to severe side effects. In addition, some checkpoint inhibitors have only moderate efficacy in clinical trials. There is still a need for improved cancer treatment, especially improved efficacy without increasing toxicity for patients. Summary of the Invention [Means for solving the problem]

[0006] overview The present disclosure encompasses the insight that combining the inhibition of checkpoint proteins with treatments that target damage to cancer cells may provide less toxic treatments with improved efficacy. Radioactive decay can cause direct physical damage (such as single- or double-stranded DNA breaks) or indirect damage (such as bystander or crossfire effects) to the biomolecules that make up cells. The present disclosure combines radioimmunoconjugates that target cancer cells with checkpoint inhibition to induce or improve immune responses against tumors. In some embodiments, the disclosed combination therapy ameliorates or treats cancer.

[0007] In one embodiment, a method of treating a patient having cancer is provided, the method comprising administering to a patient a compound of formula: AL 1 -XL 2 -ZB (wherein A is DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTMA (1R,4R,7R,10R)-α,α',α",α"'-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane), DO3AM-acetic acid (2-(4,7,10-tris(2-amino-2-oxoethyl)-1,4,7,10-triphenylphosphine)), a chelating moiety selected from 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonic acid), DOTP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonic acid)), DOTA-4AMP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(acetamido-methylenephosphonic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), and HP-DO3A (hydroxypropyltetraazacyclododecanetriacetic acid); L 1 is replaced by C 1~6 Alkyl or C1~6 is heteroalkyl, X is C=O(NR 1 ), N.R. 1 C=O(O), NR 1 C=O(NR 1 ), CH2PhC=O(NR 1 ), O, or NR 1 And each R 1 are independently H or C 1~6 is alkyl, L 2 is replaced by C 1~50 Alkyl or C 1~50 is heteroalkyl, Z is C=O, CH2, OC=O, NR 2 C=O, or NR 2 And each R 2 are independently H or C 1~6 is alkyl, B is a targeting moiety) 225 A therapeutically effective amount of [ 225 administering to the patient a .Ac]-radioimmunoconjugate; Patients have received or have received one or more checkpoint inhibitors and 225 The [Ac] radioimmunoconjugate is administered at a dose of about 10 kBq to about 400 kBq per kg of body weight of the patient, or as a unit dose of about 1 to 30 MBq to the patient.

[0008] In some embodiments, the compound has a chelating moiety that is DOTA.

[0009] In some embodiments, the compound has Formula I: [ka] has.

[0010] In some embodiments, the compound has Formula II: [ka] has.

[0011] In some embodiments, the targeting moiety comprises an antibody or an antigen-binding fragment thereof.

[0012] In some embodiments, B is an insulin-like growth factor 1 receptor (IGF-1R) antibody or an antigen-binding fragment thereof, an endosialin (TEM-1) antibody or an antigen-binding fragment thereof, or a fibroblast growth factor receptor 3 (FGFR3) antibody or an antigen-binding fragment thereof.

[0013] In some embodiments, B is an IGF-1R antibody or antigen-binding fragment thereof selected from the group consisting of figitumumab, cixutumumab, TAB-199, AVE1642, BIIB002, lobatumumab, and teprotumumab, and antigen-binding fragments thereof.

[0014] In some embodiments, B is AVE1642 or an antigen-binding fragment thereof.

[0015] In some embodiments, 225 Ac]-radioimmunoconjugate is administered at a dose of about 10 kBq to about 200 kBq per kg of body weight of the patient (e.g., about 10 kBq to about 150 kBq / kg, about 10 kBq to about 120 kBq / kg, about 10 kBq to about 100 kBq / kg, about 30 kBq to about 150 kBq / kg, about 30 kBq to about 120 kBq / kg, about 30 kBq to about 100 kBq / kg, about 40 kBq to about 120 kBq / kg, about 40 kBq to about 100 kBq / kg, or about 40 kBq to about 80 kBq / kg).

[0016] In some embodiments, [ 225 Ac]-radioimmunoconjugate is administered to the patient in a unit dose of about 1-30 MBq (eg, about 2-25 MBq, about 3-20 MBq, about 5-15 MBq, about 8-12 MBq, or about 10 MBq).

[0017] In some embodiments, the one or more checkpoint inhibitors comprise a PD-1 inhibitor, a CTLA-4 inhibitor, or a combination thereof.

[0018] In some embodiments, the one or more checkpoint inhibitors include both a PD-1 inhibitor and a CTLA-4 inhibitor.

[0019] In some embodiments, the PD-1 inhibitor or CTLA-4 inhibitor is an antibody.

[0020] In some embodiments, the one or more checkpoint inhibitors include a PD-1 inhibitor administered at a dose of about 5 mg / kg to about 15 mg / kg.

[0021] In some embodiments, the PD-1 inhibitor is pembrolizumab.

[0022] In some embodiments, the one or more checkpoint inhibitors include both a PD-1 inhibitor and a CTLA-4 inhibitor, each administered at a dose of about 5 mg / kg to about 15 mg / kg (e.g., about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, or about 14 mg / kg).

[0023] In some embodiments, B is AVE1642 or an antigen-binding fragment thereof, and the one or more checkpoint inhibitors include a PD-1 inhibitor that is pembrolizumab.

[0024] In some embodiments, [ 225 Ac] radioimmunoconjugate is administered at a dose of about 30 kBq / kg to about 120 kBq / kg of the patient's body weight, and the PD-1 inhibitor is administered at a dose of about 5 mg / kg to about 15 mg / kg.

[0025] In some embodiments, the patient has a cancer selected from the group consisting of breast cancer (e.g., triple-negative breast cancer or TNBC), non-small cell lung cancer, small cell lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, colorectal cancer, cervical cancer, endometrial cancer, sarcoma, adrenocortical carcinoma, neuroendocrine carcinoma, Ewing's sarcoma, multiple myeloma, and acute myeloid leukemia.

[0026] In some embodiments, the patient has a solid tumor that expresses IGF-1R.

[0027] In some embodiments, B is capable of binding to a tumor-associated antigen, and the administration results in an expansion of CD8+ T cells specific for the tumor-associated antigen.

[0028] In some embodiments, the administration results in at least 60% of the total CD8+ T cell population in a sample from the patient being specific for the tumor-associated antigen, hi some embodiments, the sample is a tumor sample. [Brief description of the drawings]

[0029] [Figure 1] Figure 1 shows relative tumor volumes in a CT26 syngeneic mouse tumor model after treatment with various checkpoint inhibitors. Relative tumor volumes at various time points after treatment initiation are shown for vehicle control, anti-PD-1 isotype control (15 mg / kg), anti-PD-1 (5 mg / kg or 15 mg / kg), anti-CTLA-4 isotype control (15 mg / kg), and anti-CTLA-4 (5 mg / kg or 15 mg / kg) treatment groups.

[0030] [Diagram 2] Figure 2 shows the biodistribution of [177Lu]-Compound B in the CT-26 syngeneic mouse tumor model. The percentage of injected dose per gram (%ID) in blood, bone, intestine, kidney and adrenal glands, liver and gallbladder, lung, spleen, tumor, and urine and bladder at 4, 24, 48, 96 and 168 hours is shown.

[0031] [Diagram 3] Figure 3 shows the enhanced efficacy of [225Ac]-Compound C in immunocompetent versus immunodeficient mice. Relative tumor volumes at various time points after treatment initiation are shown for control and treatment groups (50 nCi or 400 nCi [225Ac]-Compound C).

[0032] [Figure 4-1] Figure 4A shows synergy between [225Ac]-Compound C and α-CTLA-4 / PD-1 treatment in the CT26 syngeneic mouse model. Relative tumor volumes at various time points after treatment initiation are shown for control (buffer) and treatment groups (anti-CTLA-4 (5 mg / kg), anti-PD-1 (5 mg / kg), 200 nCi[225Ac]-Compound C, 200 nCi[225Ac]-Compound C with anti-CTLA-4, 200 nCi[225Ac]-Compound C with anti-PD-1, or 200 nCi[225Ac]-Compound C with anti-CTLA-4 and anti-PD-1).

[0033] [Figure 4-2] Figure 4B shows synergy between [225Ac]-Compound D and α-CTLA-4 / PD-1 treatment in the CT26 syngeneic mouse model. Relative tumor volumes at various time points after treatment initiation are shown for control (vehicle or cold human IGF-1R antibody) and treatment groups (anti-CTLA-4 (5 mg / kg), anti-PD-1 (5 mg / kg), 200 nCi[225Ac]-Compound C, 200 nCi[225Ac]-Compound C with anti-CTLA-4, 200 nCi[225Ac]-Compound C with anti-PD-1, or 200 nCi[225Ac]-Compound C with anti-CTLA-4 and anti-PD-1).

[0034] [Diagram 5]Figure 5 shows the development of protective immunity in [225Ac]-Compound C treated mice upon CT26 rechallenge. Relative tumor volumes at various time points after rechallenge are shown for control and treatment groups ([225Ac]-Compound C, [225Ac]-Compound C with anti-PD-1, [225Ac]-Compound C with anti-CTLA-4, or [225Ac]-Compound C with anti-CTLA-4 and anti-PD-1).

[0035] [Figure 6] FIG. 6 shows the process for assessing cytokine responses and T cell recruitment following [225Ac]-Compound C treatment.

[0036] [Figure 7] Figure 7 shows the development of a "humanized" IGF-1R model. Shown is a Western blot probed for expression of hIGF-1R in samples derived from CT26 cells stably transfected with the human IGF-1R plasmid.

[0037]

[0038] [Figure 8] Figure 8A is a schematic diagram showing the general structure of a bifunctional chelate comprising a chelate, a linker, and a bridging group. Figure 8B is a schematic diagram showing the general structure of a bifunctional conjugate comprising a chelate, a linker, and a targeting moiety.

[0039] [Figure 9-1] Figure 9A shows synergy between [225Ac]-Compound D1 and α-CTLA-4 / PD-1 treatment in the CT26 syngeneic mouse model. Relative tumor volumes at various time points after treatment initiation are shown for control (vehicle) and treatment groups (200 nCi [225Ac]-Compound D1, 200 nCi [225Ac]-Compound D1 with anti-PD-1, 200 nCi [225Ac]-Compound D1 with anti-CTLA-4, or 200 nCi [225Ac]-Compound C with anti-CTLA-4 and anti-PD-1).

[0040] [Figure 9-2] Figure 9B shows synergy between [225Ac]-Compound D2 and α-CTLA-4 / PD-1 treatment in the CT26 syngeneic mouse model. Relative tumor volumes at various time points after treatment initiation are shown for control (vehicle) and treatment groups (200 nCi [225Ac]-Compound D2, 200 nCi [225Ac]-Compound D2 with anti-PD-1, 200 nCi [225Ac]-Compound D2 with anti-CTLA-4, or 200 nCi [225Ac]-Compound D2 with anti-CTLA-4 and anti-PD-1). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0041] It should be understood that the drawings are not necessarily drawn to scale, and that objects in the drawings are not necessarily drawn to scale relative to each other. The drawings are representations intended to clarify and understand various embodiments of the apparatus, systems, and methods disclosed herein. Wherever possible, the same reference numbers are used throughout the drawings to refer to the same or similar parts. Furthermore, it should be understood that the drawings are not intended to limit the scope of the present teachings in any way.

[0042] Detailed Description This disclosure is 225 The present invention relates to a combination therapy for inducing or improving an immune response against cancer using a radioimmunoconjugate and a checkpoint inhibitor. In some embodiments, the use of the methods disclosed herein results in the treatment or improvement of cancer.

[0043] In some embodiments, a lower effective dose of [ 225 Ac] radioimmunoconjugates and / or checkpoint inhibitors are used.

[0044] Radiolabeled targeting moieties (also known as radioimmunoconjugates) are designed to target proteins or receptors that are upregulated in disease states and / or specific to diseased cells (e.g., tumor cells) to deliver radioactive payloads to damage and kill the cells of interest. As used herein, "radioimmunotherapy" refers to methods that use radioimmunoconjugates, such as those described below, to produce a therapeutic effect. Radioactive decay of the payload generates alpha, beta, or gamma particles or Auger electrons that can cause direct effects on DNA (such as single- or double-stranded DNA breaks) or indirect effects, such as bystander or crossfire effects.

[0045] Radioimmunoconjugates typically contain a targeting moiety (e.g., an antibody or antigen-binding fragment thereof, peptide or small molecule that specifically binds to a molecule expressed on or by a tumor, such as IGF-1R, FGFR3 or TEM-1 / endosialin), a chelating moiety or a metal complex of a chelating moiety (e.g., containing a radioisotope), and a linker. Conjugates can be formed by appending a bifunctional chelate to the targeting molecule such that structural changes are minimized while maintaining target affinity. Radioimmunoconjugates can be formed by radiolabeling such conjugates.

[0046] Bifunctional chelates structurally comprise chelate, linker and bridging group.Some examples of bifunctional chelates have been described using various cyclic and acyclic structures conjugated to targeting moiety [Bioconjugate Chem.2000,11,510-519; Bioconjugate Chem.2012,23,1029-1039; Mol Imaging Biol.2011,13,215-221; Bioconjugate Chem.2002,13,110-115]. definition chemical terms

[0047] definition Chemical terms: The term "acyl" as used herein refers to a hydrogen or alkyl group (e.g., haloalkyl group), as defined herein, attached to the parent molecular group through a carbonyl group, as defined herein, and is exemplified by formyl (i.e., carboxaldehyde group), acetyl, trifluoroacetyl, propionyl, butanoyl, and the like. Exemplary unsubstituted acyl groups contain 1 to 7, 1 to 11, or 1 to 21 carbons. In some embodiments, the alkyl group is further substituted with 1, 2, 3, or 4 substituents, as described herein.

[0048] The term "alkyl," as used herein, unless otherwise indicated, includes both straight and branched chain saturated groups of 1 to 20 carbons (e.g., 1 to 10 or 1 to 6). Alkyl groups are exemplified by methyl, ethyl, n- and iso-propyl, n-, sec-, iso- and tert-butyl, neopentyl, and the like, and may be optionally substituted with one, two, three, or, in the case of alkyl groups having two or more carbon atoms, four, substituents independently selected from the group consisting of: (1) C 1-6 Alkoxy; (2) C 1-6 (3) amino, as defined herein (e.g., unsubstituted amino (i.e., -NH) or substituted amino (i.e., -N(R N1 )2, where R N1 is as defined for amino; (4) C 6-10 Aryl-C 1-6 Alkoxy; (5) Azido; (6) Halo; (7) (C 2-9 (8) hydroxy optionally substituted with an O-protecting group; (9) nitro; (10) oxo (e.g., carboxaldehyde or acyl); (11) C 1-7 Spirocyclyl; (12) Thioalkoxy; (13) Thiol; (14) -COR optionally substituted with an O-protecting group A’ , where R A’ (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 alk-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (15) -C(O)NR B’ R C’ , where R B’ and R C’ each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 aryl; (16) -SO2R D’ , where R D’ (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 alk-C 6-10 (17) -SONR E’ R F’ , where R E’and R F’ each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 Aryl and (d) C 1-6 alk-C 6-10 aryl; (18) -C(O)R G’ , where R G’ (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 alk-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (19) -NR H’ C(O)R I’ , where R H’ is (a1) hydrogen and (b1) C 1-6 alkyl; R I’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 alk-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h2) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (20) -NR J’ C(O)OR K’ , where R J’ is (a1) hydrogen and (b1) C 1-6 alkyl; R K’ is (a2)C 1-20 Alkyl (e.g., C 1-6 (b2) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 alk-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3OR' polyethylene glycol, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h2) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 and (21) amidine. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of C1-alkaryl can be further substituted with an oxo group to provide the respective aryloyl substituent.

[0049] The terms "alkylene" and the prefix "alk-" as used herein refer to a saturated divalent hydrocarbon group derived from a straight or branched chain saturated hydrocarbon by the removal of two hydrogen atoms, and are exemplified by methylene, ethylene, isopropylene, and the like. x-y Alkylene" and the prefix "C x-y "alk-" represents an alkylene group having x to y carbons. Exemplary values ​​of x are 1, 2, 3, 4, 5, and 6, and exemplary values ​​of y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C 1-6 , C 1-10 , C 2-20 , C 2-6 , C 2-10 , or C 2-20 In some embodiments, the alkylene can be further substituted with 1, 2, 3, or 4 substituents as defined herein for an alkyl group.

[0050] The term "alkenyl", unless otherwise indicated, refers to a monovalent straight or branched chain group of 2 to 20 carbons (e.g., 2 to 6 or 2 to 10 carbons) containing one or more carbon-carbon double bonds, exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like. Alkenyl includes both cis and trans isomers. Alkenyl groups may be optionally substituted with 1, 2, 3, or 4 substituents independently selected from amino, aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituents described herein.

[0051] The term "alkynyl" as used herein refers to a monovalent straight or branched chain group of 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbons) containing a carbon-carbon triple bond, exemplified by ethynyl, 1-propynyl, etc. Alkynyl groups can be optionally substituted with 1, 2, 3, or 4 substituents independently selected from aryl, cycloalkyl, or heterocyclyl (e.g., heteroaryl), as defined herein, or any of the exemplary alkyl substituents described herein.

[0052] As used herein, the term “amino” refers to —N(R N1 )2, wherein each R N1 are independently H, OH, NO2, and N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2, an N-protecting group, an alkyl, an alkenyl, an alkynyl, an alkoxy, an aryl, an alkaryl, a cycloalkyl, an alkcycloalkyl, a carboxyalkyl (e.g., optionally substituted with an O-protecting group, e.g., optionally substituted with an arylalkoxycarbonyl group or any described herein), a sulfoalkyl, an acyl (e.g., acetyl, trifluoroacetyl, or others described herein), an alkoxycarbonylalkyl (e.g., optionally substituted with an O-protecting group, e.g., optionally substituted with an arylalkoxycarbonyl group or any described herein), a heterocyclyl (e.g., heteroaryl), or an alkheterocyclyl (e.g., alkheteroaryl), where these enumerated R N1 Each of the groups may be optionally substituted as defined herein for each group; or two R N1 can be linked to form a heterocyclyl or an N-protecting group, N2 is independently H, alkyl, or aryl. The amino group may be an unsubstituted amino (i.e., -NH) group or a substituted amino (i.e., -N(R N1 )2) group. In a preferred embodiment, amino is -NH2 or -NHR N1 where R N1 are independently OH, NO2, NH2, and NR N2 2. SO2OR N2 , SO2R N2 , SOR N2 , alkyl, carboxyalkyl, sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., t-butoxycarbonylalkyl), or aryl, and each R N2 , H, C 1-20 Alkyl (e.g., C 1-6 alkyl), or C 6-10 It may be aryl.

[0053] As described herein, an "amino acid" refers to a molecule having a side chain, an amino group, and an acid group (e.g., a carboxy group of -CO2H or a sulfo group of -SO3H), where the amino acid is attached to the parent molecular group by the side chain, the amino group, or the acid group (e.g., the side chain). In some embodiments, the amino acid is attached to the parent molecular group by a carbonyl group, where the side chain or the amino group is attached to the carbonyl group. Exemplary side chains include optionally substituted alkyl, aryl, heterocyclyl, alkaryl, alkheterocyclyl, aminoalkyl, carbamoylalkyl, and carboxyalkyl. Exemplary amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, hydroxynorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolysine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine. The amino acid group may be optionally substituted with one, two, three, or, in the case of amino acid groups having two or more carbons, four, substituents independently selected from the group consisting of: (1) C 1-6 Alkoxy; (2) C 1-6 (3) amino, as defined herein (e.g., unsubstituted amino (i.e., -NH) or substituted amino (i.e., -N(R N1 )2, where R N1 is as defined for amino; (4) C 6-10 Aryl-C 1-6 Alkoxy; (5) Azido; (6) Halo; (7) (C 2-9 (8) hydroxy; (9) nitro; (10) oxo (e.g., carboxaldehyde or acyl); (11) C 1-7 Spirocyclyl; (12) Thioalkoxy; (13) Thiol; (14) -CO2R A’ , where R A’ (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 alk-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (15) -C(O)NR B’ R C’ , where R B’ and R C’ each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 aryl; (16) -SO2R D’ , where R D’ (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 alk-C 6-10 (17) -SONR E’ R F’ , where R E’ and R F’each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 Aryl and (d) C 1-6 alk-C 6-10 aryl; (18) -C(O)R G’ , where R G’ (a)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c) C 6-10 Aryl, (d) hydrogen, (e) C 1-6 alk-C 6-10 Aryl, (f) Amino-C 1-20 Alkyl, (g)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (19) -NR H’ C(O)R I’ , where R H’ is (a1) hydrogen and (b1) C 1-6 alkyl; R I’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C 2-20Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 alk-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3 OR' polyethylene glycol, where s1 is an integer of 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer of 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h2) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 (20) -NR J’ C(O)OR K’ , where R J’ is (a1) hydrogen and (b1) C 1-6 alkyl; R K’ is (a2)C 1-20 Alkyl (e.g., C 1-6 alkyl), (b2) C 2-20 Alkenyl (e.g., C 2-6 alkenyl), (c2) C 6-10 Aryl, (d2) hydrogen, (e2) C 1-6 alk-C 6-10 Aryl, (f2) Amino-C 1-20 Alkyl, (g2)-(CH2) s2 (OCH2CH2) s1 (CH2) s3OR' polyethylene glycol, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), s2 and s3 are each independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and R' is H or C 1-20 alkyl), and (h2) -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1 wherein s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 and (21) amidine. In some embodiments, each of these groups can be further substituted as described herein.

[0054] The term "aryl," as used herein, refers to a monocyclic, bicyclic, or polycyclic carbocyclic ring system having one or two aromatic rings, exemplified by phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, anthracenyl, phenanthrenyl, fluorenyl, indanyl, indenyl, and the like, optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6 Alkyl, (Carboxaldehyde)-C 1-6 Alkyl, halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, for example perfluoroalkoxy; (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 alk-C 6-10 Aryl;(8)Azide;(9)C 3-8 Cycloalkyl; (10)C 1-6 alk-C 3-8 Cycloalkyl;(11)Halo;(12)C 1-12 Heterocyclyl (e.g., C 1-12 Heteroaryl; (13) (C 1-12 (14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6 Thioalkoxy;(17)-(CH2) q CO2R A’ (where q is an integer from 0 to 4, and R A’ (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 alk-C 6-10 aryl);(18)-(CH2) q CONR B’ R C’ where q is an integer from 0 to 4, and R B’ and R C’ are independently: (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 aryl);(19)-(CH2) q SO2R D’ (where q is an integer from 0 to 4, and R D’ is (a) alkyl, (b) C 6-10 aryl, and (c) alk-C 6-10aryl);(20)-(CH2) q SO2NR E’ R F’ (where q is an integer from 0 to 4, and R E’ and R F’ each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C 3-8 Cycloalkoxy; (24)C 6-10 Aryl-C 1-6 Alkoxy; (25)C 1-6 alk-C 1-12 Heterocyclyl (e.g., C 1-6 alk-C 1-12 Heteroaryl;(26)C 2-20 alkenyl; and (27) C 2-20 Alkynyl. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to obtain the respective aryloyl and (heterocyclyl)oyl substituents.

[0055] The term "arylalkyl" as used herein refers to an aryl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein. Exemplary unsubstituted arylalkyl groups include those having 7 to 30 carbons (e.g., 7 to 16 or 7 to 20 carbons, e.g., C 1-6 alk-C 6-10 Aryl, C 1-10 alk-C 6-10 Aryl, or C 1-20 alk-C 6-10In some embodiments, alkylene and aryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the respective group. Other groups preceded by the prefix "alk-" are similarly defined, where "alk" is any of the groups listed below, unless otherwise specified. 1-6 Refers to alkylene and the chemical structures to which it is attached are as defined herein.

[0056] The term "carbonyl" as used herein refers to a C(O) group, which may also be represented as C=O.

[0057] The term "carboxy" as used herein means -CO2H.

[0058] As used herein, the term "cyano" refers to a -CN group.

[0059] As used herein, unless otherwise indicated, "cycloalkyl" refers to a monovalent saturated or unsaturated non-aromatic cyclic hydrocarbon group of 3 to 8 carbons, exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, bicyclo heptyl, and the like. When a cycloalkyl group contains one carbon-carbon double bond or one carbon-carbon triple bond, the cycloalkyl group can be referred to as a "cycloalkenyl" or "cycloalkynyl" group, respectively. Exemplary cycloalkenyl and cycloalkynyl groups include cyclopentenyl, cyclohexenyl, cyclohexynyl, and the like. Cycloalkyl groups can be optionally substituted with: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6Alkyl, (Carboxaldehyde)-C 1-6 Alkyl, halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, for example perfluoroalkoxy; (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 alk-C 6-10 Aryl;(8)Azide;(9)C 3-8 Cycloalkyl; (10)C 1-6 alk-C 3-8 Cycloalkyl;(11)Halo;(12)C 1-12 Heterocyclyl (e.g., C 1-12 Heteroaryl; (13) (C 1-12 (14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6 Thioalkoxy;(17)-(CH2) q CO2R A’ (where q is an integer from 0 to 4, and R A’ (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 alk-C 6-10 aryl);(18)-(CH2) q CONR B’ R C’ where q is an integer from 0 to 4, and R B’ and R C’ are independently: (a) hydrogen, (b) C 6-10 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 aryl);(19)-(CH2) q SO2R D’(where q is an integer from 0 to 4, and R D’ (a)C 6-10 Alkyl, (b) C 6-10 aryl, and (c) C 1-6 alk-C 6-10 aryl);(20)-(CH2) q SO2NR E’ R F’ (where q is an integer from 0 to 4, and R E’ and R F’ each independently is (a) hydrogen, (b) C 6-10 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C 3-8 Cycloalkoxy; (24)C 6-10 Aryl-C 1-6 Alkoxy; (25)C 1-6 alk-C 1-12 Heterocyclyl (e.g., C 1-6 alk-C 1-12 Heteroaryl;(26)oxo;(27)C 2-20 alkenyl; and (28) C 2-20 Alkynyl. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to obtain the respective aryloyl and (heterocyclyl)oyl substituents.

[0060] The term "diastereomers" as used herein means stereoisomers which are not mirror images of one another and are not superimposable with respect to one another.

[0061] As used herein, the term "enantiomer" refers to each individual optically active form of a compound having an optical purity or enantiomeric excess (as measured by standard methods in the art) of at least 80% (i.e., at least 90% of one enantiomer and up to 10% of the other enantiomer), preferably at least 90%, and more preferably at least 98%.

[0062] The term "halogen" as used herein refers to a halogen selected from bromine, chlorine, iodine, or fluorine.

[0063] The term "heteroalkyl" as used herein refers to an alkyl group as defined herein, in which one or two of the constituent carbon atoms are replaced with nitrogen, oxygen, or sulfur, respectively. In some embodiments, the heteroalkyl group can be further substituted with one, two, three, or four substituents as described herein for alkyl groups. The terms "heteroalkenyl" and "heteroalkynyl" as used herein refer to an alkenyl group and an alkynyl group, as defined herein, in which one or two of the constituent carbon atoms are replaced with nitrogen, oxygen, or sulfur, respectively. In some embodiments, the heteroalkenyl group and the heteroalkynyl group can be further substituted with one, two, three, or four substituents as described herein for alkyl groups.

[0064] The term "heteroaryl" as used herein refers to a subset of heterocyclyl, as defined herein, that are aromatic, i.e., they contain 4n+2 pi-electrons in a monocyclic or polycyclic ring system. Exemplary unsubstituted heteroaryl groups are those of 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbons. In some embodiments, heteroaryl is substituted with 1, 2, 3, or 4 substituents as defined for heterocyclyl groups.

[0065] As used herein, "heteroarylalkyl" refers to a heteroaryl group, as defined herein, attached to the parent molecular group through an alkylene group, as defined herein. Exemplary unsubstituted heteroarylalkyl groups include those having 2 to 32 carbons (e.g., 2 to 22, 2 to 18, 2 to 17, 2 to 16, 3 to 15, 2 to 14, 2 to 13, or 2 to 12 carbons, such as C 1-6 alk-C 1-12 Heteroaryl, C 1-10 alk-C 1-12 Heteroaryl, or C 1-20 alk-C 1-12 In some embodiments, alkylene and heteroaryl can each be further substituted with 1, 2, 3, or 4 substituents as defined herein for the respective group. Heteroarylalkyl groups are a subset of heterocyclylalkyl groups.

[0066] The term "heterocyclyl" as used herein, unless otherwise indicated, refers to a 5-, 6-, or 7-membered ring containing 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Five-membered rings have 0-2 double bonds, and 6- and 7-membered rings have 0-3 double bonds. Exemplary unsubstituted heterocyclyl groups are those of 1-12 (e.g., 1-11, 1-10, 1-9, 2-12, 2-11, 2-10, or 2-9) carbons. The term "heterocyclyl" also refers to heterocyclic compounds having bridged polycyclic structures in which one or more carbons and / or heteroatoms bridge two non-adjacent members of a monocyclic ring, e.g., a quinuclidinyl group. The term "heterocyclyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocycles are fused to one, two, or three carbocyclic rings, such as an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocycle, such as indolyl, quinolyl, isoquinolyl, tetrahydroquinolyl, benzofuryl, benzothienyl, etc. Examples of fused heterocyclyls include tropane and 1,2,3,5,8,8a-hexahydroindolizine. Heterocycles include pyrrolyl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl, homopiperidinyl, pyrazinyl, piperazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl, isoxazolyl, isoxazolidinyl, morpholinyl, thiomorpholinyl, thiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, nyl, indolyl, indazolyl, quinolyl, isoquinolyl, quinoxalinyl, dihydroquinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzimidazolyl, benzothiazolyl, benzoxazolyl, benzothiadiazolyl, furyl, thienyl, thiazolidinyl, isothiazolyl, triazolyl, tetrazolyl, oxadiazolyl (e.g., 1,2,3-oxadiazolyl), purinyl, thiadiazolyl (e.g., 1,2,3-thiadiazolyl), tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, dihydroindolyl, dihydroquinolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, dihydroisoquinolyl, pyranyl, dihydropyranyl, dithiazolyl, benzofuranyl, isobenzofuranyl, benzothienyl, and the like, including dihydro and tetrahydro forms thereof in which one or more double bonds have been reduced and replaced with hydrogen. Still other exemplary heterocyclyls include: 2,3,4,5-tetrahydro-2-oxo-oxazolyl; 2,3-dihydro-2-oxo-1H-imidazolyl; 2,3,4,5-tetrahydro-5-oxo-1H-pyrazolyl (e.g., 2,3,4,5-tetrahydro-2-phenyl-5-oxo-1H-pyrazolyl); 2,3,4,5-tetrahydro-2,4-dioxo-1H-imidazolyl (e.g., , 2,3,4,5-tetrahydro-2,4-dioxo-5-methyl-5-phenyl-1H-imidazolyl; 2,3-dihydro-2-thioxo-1,3,4-oxadiazolyl (e.g., 2,3-dihydro-2-thioxo-5-phenyl-1,3,4-oxadiazolyl); 4,5-dihydro-5-oxo-1H-triazolyl (e.g., 4,5-dihydro-3-methyl-4-amino 5-oxo-1H-triazolyl); 1, 2,3,4-tetrahydro-2,4-dioxopyridinyl (e.g., 1,2,3,4-tetrahydro-2,4-dioxo-3,3-diethylpyridinyl); 2,6-dioxo-piperidinyl (e.g., 2,6-dioxo-3-ethyl-3-phenylpiperidinyl); 1,6-dihydro-6-oxopyrimidinyl; 1,6-dihydro-4-oxopyrimidinyl (e.g., 2-(methylthio)- 1,6-dihydro-4-oxo-5-methylpyrimidin-1-yl; 1,2,3,4-tetrahydro-2,4-dioxopyrimidinyl (e.g., 1,2,3,4-tetrahydro-2,4-dioxo-3-ethylpyrimidinyl); 1,6-dihydro-6-oxo-pyridazinyl (e.g., 1,6-dihydro-6-oxo-3-ethylpyridazinyl); 1,6-dihydro-6-oxo-1,2,4-triazinyl (e.g., 1,6-dihydro-5-isopropyl-6-oxo-1,2,4-triazinyl);2,3-dihydro-2-oxo-1H-indolyl (e.g., 3,3-dimethyl-2,3-dihydro-2-oxo-1H-indolyl and 2,3-dihydro-2-oxo-3,3'-spiropropane-1H-indol-1-yl);1,3-dihydro-1-oxo-2H-iso-indolyl;1,3-dihydro-1,3-dioxo-2H-iso-indolyl;1H-benzopyra 2,3-dihydro-2-oxo-1H-benzimidazolyl (e.g., 3-ethyl-2,3-dihydro-2-oxo-1H-benzimidazolyl); 2,3-dihydro-2-oxo-benzoxazolyl (e.g., 5-chloro-2,3-dihydro-2-oxo-benzoxazolyl); 2,3-dihydro-2-oxo-benzoxazolyl; 2-oxo-2H-benzopyranyl; 1,4-benzo Dioxanyl; 1,3-benzodioxanyl; 2,3-dihydro-3-oxo,4H-1,3-benzothiazinyl; 3,4-dihydro-4-oxo-3H-quinazolinyl (e.g., 2-methyl-3,4-dihydro-4-oxo-3H-quinazolinyl); 1,2,3,4-tetrahydro-2,4-dioxo-3H-quinazolyl (e.g., 1-ethyl-1,2,3,4-tetrahydro-2,4-dioxo-3H-quinazolyl); 1,2,3,6-tetrahydro-2,6- Dioxo-7H-purinyl (e.g., 1,2,3,6-tetrahydro-1,3-dimethyl-2,6-dioxo-7H-purinyl); 1,2,3,6-tetrahydro-2,6-dioxo-1H-purinyl (e.g., 1,2,3,6-tetrahydro-3,7-dimethyl-2,6-dioxo-1H-purinyl); 2-oxobenzo[c,d]indolyl; 1,1-dioxo-2H-naphtho[1,8-c,d]isothiazolyl; and 1,8-naphthylenedicarboxamide. Further heterocycles include 3,3a,4,5,6,6a-hexahydro-pyrrolo[3,4-b]pyrrol-(2H)-yl, and 2,Heterocyclic groups include 5-diazabicyclo[2.2.1]heptan-2-yl, homopiperazinyl (or diazepanyl), tetrahydropyranyl, dithiazolyl, benzofuranyl, benzothienyl, oxepanyl, thiepanyl, azocanyl, oxecanyl, and thiocanyl. Heterocyclic groups also include groups of the formula: [ka] Where: E' is selected from the group consisting of -N- and -CH-; F' is selected from the group consisting of -N=CH-, -NH-CH2-, -NH-C(O)-, -NH-, -CH=N-, -CH2-NH-, -C(O)-NH-, -CH=CH-, -CH2-, -CH2CH2-, -CHO-, -OCH2-, -O-, and -S-; G' is selected from the group consisting of -CH- and -N-. Any heterocyclyl group referred to herein may be optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of: (1) C 1-7 Acyl (e.g., carboxaldehyde); (2) C 1-20 Alkyl (e.g., C 1-6 Alkyl, C 1-6 Alkoxy-C 1-6 Alkyl, C 1-6 Alkylsulfinyl-C 1-6 Alkyl, Amino-C 1-6 Alkyl, azido-C 1-6 Alkyl, (Carboxaldehyde)-C 1-6 Alkyl, halo-C 1-6 Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, for example perfluoroalkoxy; (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6 alk-C6-10 Aryl;(8)Azide;(9)C 3-8 Cycloalkyl; (10)C 1-6 alk-C 3-8 Cycloalkyl;(11)Halo;(12)C 1-12 Heterocyclyl (e.g., C 2-12 Heteroaryl; (13) (C 1-12 (14) Hydroxy; (15) Nitro; (16) C 1-20 Thioalkoxy (e.g., C 1-6 Thioalkoxy;(17)-(CH2) q CO2R A’ (where q is an integer from 0 to 4, and R A’ (a)C 1-6 Alkyl, (b) C 6-10 aryl, (c) hydrogen, and (d) C 1-6 alk-C 6-10 aryl);(18)-(CH2) q CONR B’ R C’ (where q is an integer from 0 to 4, and R B’ and R C’ are independently: (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 aryl);(19)-(CH2) q SO2R D’ (where q is an integer from 0 to 4, and R D’ (a)C 1-6 Alkyl, (b) C 6-10 aryl, and (c) C 1-6 alk-C 6-10 aryl);(20)-(CH2) q SO2NR E’ R F’ (where q is an integer from 0 to 4, and R E’ and R F’ each independently is (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6alk-C 6-10 (21) thiol; (22) C 6-10 Aryloxy; (23)C 3-8 Cycloalkoxy; (24) Arylalkoxy; (25) C 1-6 alk-C 1-12 Heterocyclyl (e.g., C 1-6 alk-C 1-12 Heteroaryl; (26) oxo; (27) (C 1-12 Heterocyclyl)imino;(28)C 2-20 alkenyl; and (29) C 2-20 Alkynyl. In some embodiments, each of these groups can be further substituted as described herein. For example, the alkylene group of C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to obtain the respective aryloyl and (heterocyclyl)oyl substituents.

[0067] As used herein, the term "hydrocarbon" refers to a group consisting solely of carbon and hydrogen atoms.

[0068] The term "hydroxyl" as used herein refers to an -OH group. In some embodiments, the hydroxyl group can be substituted with 1, 2, 3, or 4 substituents (e.g., O-protecting groups) as defined herein for alkyl.

[0069] The term "isomer" as used herein refers to any tautomer, stereoisomer, enantiomer, or diastereomer of any compound. It is recognized that compounds can have one or more chiral centers and / or double bonds and therefore can exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). Unless otherwise indicated, chemical structures depicted herein encompass all corresponding stereoisomers, i.e., both stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers, e.g., racemates. Enantiomeric and stereoisomeric mixtures of a compound can generally be resolved into its component enantiomers or stereoisomers by well-known methods, such as chiral-phase gas chromatography, chiral-phase high performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent, etc. Enantiomers and stereoisomers can also be obtained by well-known asymmetric synthetic methods from stereomerically or enantiomerically pure intermediates, reagents, and catalysts.

[0070] The term "N-protected amino" as used herein refers to an amino group, as defined herein, having attached thereto one or two N-protecting groups, as defined herein.

[0071] The term "N-protecting group" as used herein refers to a group intended to protect an amino group from undesired reactions during synthetic procedures. Commonly used N-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis" 3rd Edition (John Wiley & Sons, New York, 1999), which is incorporated herein by reference.N-protecting groups include acyl, aryloyl or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like, and protecting or isopropyl groups such as chiral auxiliaries, such as alanine, leucine, phenylalanine, and the like. unprotected D,L or D,L-amino acids; sulfonyl-containing groups such as benzenesulfonyl, p-toluenesulfonyl; carbamate-forming groups, such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethylbenzyloxycarbonyl, ethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl These include aryl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, etc., alkaryl groups such as benzyl, triphenylmethyl, benzyloxymethyl, etc., and silyl groups such as trimethylsilyl, etc. Preferred N-protecting groups are formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0072] The term "O-protecting group" as used herein refers to a group intended to protect an oxygen-containing (e.g., phenol, hydroxyl, or carbonyl) group from undesired reactions during synthetic procedures. Commonly used O-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis" 3rd Edition (John Wiley & Sons, New York, 1999), which is incorporated herein by reference. Exemplary O-protecting groups include acyl, aryloyl or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, t-butyldimethylsilyl, tri-iso-propylsilyloxymethyl, 4,4′-dimethoxytrityl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidino, and 4-nitrobenzoyl; alkylcarbonyl groups, such as acyl, acetyl, propionyl, pivaloyl, and the like; and, where appropriate, substituted arylcarbonyl groups, such as benzoyl; silyl groups, such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-isopropylsilyloxymethyl (TOM), triisopropylsilyl (TIPS), and the like; hydroxyl-containing ether-forming groups, such as methyl, methoxymethyl, tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl, and the like; alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, n-isopropoxycarbonyl, n-butyloxycarbonyl, isobutyloxycarbonyl, sec-butyloxycarbonyl, t-butyloxycarbonyl, 2-ethylhexyloxycarbonyl, cyclohexyloxycarbonyl, methyloxycarbonyl, and the like;Alkoxyalkoxycarbonyl groups, such as methoxymethoxycarbonyl, ethoxymethoxycarbonyl, 2-methoxyethoxycarbonyl, 2-ethoxyethoxycarbonyl, 2-butoxyethoxycarbonyl, 2-methoxyethoxymethoxycarbonyl, allyloxycarbonyl, propargyloxycarbonyl, 2-butenoxycarbonyl, 3-methyl-2-butenoxycarbonyl, etc.; haloalkoxycarbonyl, such as 2-chloroethoxycarbonyl, 2-chloroethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, etc. optionally substituted arylalkoxycarbonyl groups, such as benzyloxycarbonyl, p-methylbenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,4-dinitrobenzyloxycarbonyl, 3,5-dimethylbenzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, fluorenylmethyloxycarbonyl, and the like; and optionally substituted aryloxycarbonyl groups, such as phenoxycarbonyl, p -nitrophenoxycarbonyl, o-nitrophenoxycarbonyl, 2,4-dinitrophenoxycarbonyl, p-methyl-phenoxycarbonyl, m-methylphenoxycarbonyl, o-bromophenoxycarbonyl, 3,5-dimethylphenoxycarbonyl, p-chlorophenoxycarbonyl, 2-chloro-4-nitrophenoxycarbonyl, etc.; substituted alkyl, aryl, and alkaryl ethers (e.g., trityl; methylthiomethyl; methoxymethyl; benzyloxymethyl; siloxymethyl; 2,2,2,-trichloroethoxy; dimethyl; tetrahydropyranyl; tetrahydrofuranyl; ethoxyethyl; 1-[2-(trimethylsilyl)ethoxy]ethyl; 2-trimethylsilylethyl; t-butyl ether; p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, benzyl, p-methoxybenzyl, and nitrobenzyl; silyl ethers (e.g., trimethylsilyl; triethylsilyl; triisopropylsilyl; dimethylisopropylsilyl; t-butyldimethylsilyl; t-butyldiphenylsilyl; tribenzylsilyl; triphenylsilyl;and diphenymethylsilyl); carbonates (e.g., methyl, methoxymethyl, 9-fluorenylmethyl; ethyl; 2,2,2-trichloroethyl; 2-(trimethylsilyl)ethyl; vinyl, allyl, nitrophenyl; benzyl; methoxybenzyl; 3,4-dimethoxybenzyl; and nitrobenzyl); carbonyl protecting groups (e.g., acetal and ketal groups, such as dimethylacetal, 1,3-dioxolane, and the like; acylal groups; and dithiane groups, such as 1,3-dithiane, 1,3-dithiolane, and the like); carboxylic acid protecting groups (e.g., ester groups, such as methyl ester, benzyl ester, t-butyl ester, ortho ester, and the like; and oxazoline groups.

[0073] The term "oxo" as used herein refers to =O.

[0074] The term "polyethylene glycol" as used herein refers to an alkoxy chain of one or more monomeric units, each of which consists of -OCH2CH2-. Polyethyelene glycol (PEG) is also sometimes called polyethylene oxide (PEO) or polyoxyethylene (POE), and these terms are considered interchangeable for purposes of this disclosure. For example, polyethylene glycol is -(CH2) s2 (OCH2CH2) s1 (CH2) s3 O-, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), and each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10). Polyethylene glycol may also have the structure -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1-amino-polyethylene glycol, where s1 is an integer from 1 to 10 (e.g., 1 to 6 or 1 to 4), each of s2 and s3 is independently an integer from 0 to 10 (e.g., 0 to 4, 0 to 6, 1 to 4, 1 to 6, or 1 to 10), and each R N1 are independently hydrogen or optionally substituted C 1-6 The term "alkyl" may also be considered to include the term "alkyl."

[0075] The term "stereoisomer" as used herein refers to all possible different isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may possess, in particular all stereochemically and conformationally possible isomers, all diastereomers, enantiomers and / or conformers of a basic molecular structure. Some compounds may exist in different tautomeric forms, and all tautomeric forms are included within the scope of this disclosure.

[0076] The term "sulfonyl" as used herein refers to the group -S(O)2-.

[0077] The term "thiol" as used herein refers to a --SH group.

[0078] Other Terms The term "about" or "approximately" as used herein refers to a ±10% variation from the recited quantitative value (including the recited quantitative value itself) unless otherwise indicated or inferred from the context. For example, unless otherwise stated or inferred from the context, a dose of about 100 kBq / kg indicates a dose range of 100±10% kBq / kg, i.e., 90 kBq / kg to 110 kBq / kg (both inclusive).

[0079] As used herein, the term "combined administration", "concurrent administration" or "co-administration" means that two or more agents are administered to a subject at the same time or at such intervals that the effects of each agent on the patient may overlap. Thus, two or more agents administered in combination do not need to be administered together. In some embodiments, they are administered within 90 days (e.g., within 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 days) of each other, within 28 days (e.g., within 14, 7, 6, 5, 4, 3, 2, or 1 days), within 24 hours (e.g., within 12, 6, 5, 4, 3, 2, or 1 hours), or within about 60, 30, 15, 10, 5, or 1 minute. In some embodiments, the administration of the agents is spaced close enough together to achieve the effect of the combination.

[0080] As used herein, "administering" an agent to a subject includes contacting cells of the subject with the agent.

[0081] As used herein, "antibody" refers to a polypeptide comprising an amino acid sequence comprising an immunoglobulin and fragments thereof that specifically binds to a designated antigen, or a fragment thereof. An antibody may be of any type (e.g., IgA, IgD, IgE, IgG, or IgM) or subtype (e.g., IgA1, IgA2, IgG1, IgG2, IgG3, or IgG4). One of skill in the art will understand that a characteristic sequence or portion of an antibody may include amino acid sequences found in one or more regions of an antibody (e.g., variable region, hypervariable region, constant region, heavy chain, light chain, and combinations thereof). Furthermore, one of skill in the art will understand that a characteristic sequence or portion of an antibody may include one or more polypeptide chains and may include sequence elements found in the same or different polypeptide chains.

[0082] As used herein, "antigen-binding fragment" refers to a portion of an antibody that retains the binding characteristics of the parent antibody.

[0083] The term "bifunctional chelate" as used herein refers to a compound that includes a chelate, a linker, and a bridging group.See, for example, Figure 8A.A "bridging group" is a reactive group that can covalently link two or more molecules, for example, link a bifunctional chelate with a targeting moiety.

[0084] As used herein, the term "bifunctional conjugate" refers to a compound that includes a chelate or a metal complex thereof, a linker, and a targeting moiety, such as an antibody or an antigen-binding fragment thereof. See, e.g., Figure 8B.

[0085] The term "cancer" refers to any cancer caused by the proliferation of malignant neoplastic cells, such as tumors, neoplasms, carcinomas, sarcomas, leukemias, and lymphomas. "Solid tumor cancers" are cancers that involve an abnormal mass of tissue, such as sarcomas, carcinomas, and lymphomas. "Blood cancers" or "liquid cancers," used interchangeably herein, are cancers that are present in bodily fluids, such as lymphomas and leukemias.

[0086] The term "checkpoint inhibitors," also known as "immune checkpoint inhibitors" or "ICIs," refers to agents that block the action of immune checkpoint proteins, e.g., that block the binding of such immune checkpoint proteins to their partner proteins.

[0087] The term "chelate" refers to an organic compound or moiety thereof that is capable of binding at two or more points to a central metal or radioactive metal atom.

[0088] As used herein, the term "conjugate" refers to a molecule that contains a chelating group or a metal complex thereof, a linker group, and optionally a therapeutic moiety, a targeting moiety, or a bridging group.

[0089] As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, and tautomers of the structures depicted.

[0090] The compounds described herein may be asymmetric (e.g., have one or more stereocenters). Unless otherwise specified, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of the present disclosure that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically active starting materials are known in the art, such as by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers of olefins, C=N double bonds, and the like, may also exist in the compounds described herein, and all such stable isomers are contemplated in the present disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described, and they may be isolated as a mixture of isomers or as separate isomeric forms.

[0091] The compounds of the present disclosure also include tautomeric forms. Tautomeric forms result from the exchange of a single bond with an adjacent double bond and the accompanying migration of a proton. Tautomeric forms include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, amide-imidic acid pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions of a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0092] At various places in the present specification, substituents of compounds of the present disclosure are disclosed in groups or in ranges. The present disclosure is specifically intended to include each and every individual subcombination of the members of such groups or ranges. For example, "C 1-6 The term "alkyl" is specifically intended to disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl individually. As used herein, phrases of the form "optionally substituted X" (e.g., optionally substituted alkyl) are intended to be equivalent to "X, where X is optionally substituted" (e.g., "alkyl, said alkyl is optionally substituted"). This does not imply that the feature "X" (e.g., alkyl) itself is optional.

[0093] As used herein, "bridging group" refers to any reactive group that can covalently link two or more molecules. In some embodiments, the bridging group is an amino-reactive or thiol-reactive bridging group. In some embodiments, the amino-reactive or thiol-reactive bridging group comprises an activated ester, such as hydroxysuccinimide ester, 2,3,5,6-tetrafluorophenol ester, 4-nitrophenol ester, or an imidate, anhydride, thiol, disulfide, maleimide, azide, alkyne, strained alkyne, strained alkene, halogen, sulfonate, haloacetyl, amine, hydrazide, diazirine, phosphine, tetrazine, isothiocyanate. In some embodiments, the bridging group can be glycine-glycine-glycine and / or leucine-proline-(any amino acid)-threonine-glycine. These are recognition sequences for coupling targeting agents and linkers using sortase-mediated coupling reactions. One of skill in the art will appreciate that the use of cross-linking groups is not limited to the specific constructs disclosed herein, but rather may include other known cross-linking groups.

[0094] As used herein (e.g., in relation to a treatment outcome or effect), the terms "reduce", "reduced", "increase", "increase", or "reduce" and "reduced" have a meaning relative to a reference level. In some embodiments, the reference level is a level determined by using the method with a control in an experimental animal model or clinical trial. In some embodiments, the reference level is a level before or at the start of treatment in the same subject. In some embodiments, the reference level is an average level in a population not treated by the treatment method.

[0095] As used herein, the term "effective amount" of an agent (e.g., any of the conjugates described above) is an amount sufficient to effect beneficial or desired results, such as clinical results, and thus, "effective amount" will vary depending on the context in which it is applied.

[0096] As used herein, the term "immunoconjugate" refers to a conjugate that includes a targeting moiety (e.g., an antibody, a nanobody, an affibody, a consensus sequence derived from a fibronectin type III domain, a peptide, or a small molecule, etc.). In some embodiments, the immunoconjugate includes an average of at least 0.10 conjugates per targeting moiety (e.g., an average of at least 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 4, 5, or 8 conjugates per targeting moiety).

[0097] As used herein, a "lower effective dose" refers to a dosage of an agent (e.g., a therapeutic agent) that is therapeutically effective in a combination therapy of the invention when used in conjunction with that agent, and that is lower than the dose that has been determined to be therapeutically effective when that agent is used as a monotherapy in a reference experiment or for other therapeutic guidance.

[0098] The term "pharmaceutical composition" refers to a composition containing a compound described herein that is formulated with a pharma-ceutical acceptable excipient. In some embodiments, the pharmaceutical composition is manufactured or sold with the approval of a government regulatory agency as part of a therapeutic regimen for the treatment of a disease in a mammal. The pharmaceutical composition can be formulated, for example, in a unit dosage form for oral administration (e.g., tablet, capsule, caplet, gelcap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution that does not contain particulate emboli and in a solvent system suitable for intravenous use); or in any other formulation described herein.

[0099] As used herein, "pharmaceutical acceptable excipient" refers to a component other than the compounds described herein (e.g., a vehicle that can suspend or dissolve active compounds) and has the characteristics of being non-toxic and non-inflammatory in patients.Excipients can include, for example, antiadherent, antioxidant, binder, coating, compression aid, disintegrant, dye (pigment), emollient, emulsifier, filler (diluent), film-forming or coating agent, flavoring agent, fragrance, glidant (flow enhancer), lubricant, preservative, printing ink, radiation protection agent, adsorbent, suspending or dispersing agent, sweetener, or water for hydration. Exemplary excipients include, but are not limited to, ascorbic acid, histidine, phosphate buffer, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0100] As used herein, "pharmaceutically acceptable salt" refers to a salt of the compound described herein that is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, or allergic reaction within the scope of sound medical judgment.Pharmaceutically acceptable salts are well known in the art.For example, pharmaceutically acceptable salts are described in Berge et al., J.Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds.P.H.Stahl and C.G.Wermuth), Wiley-VCH, 2008.Salts can be prepared in situ during the final isolation and purification of the compound described herein, or separately by reacting free base group with suitable organic acid.

[0101] Compounds may have ionizable groups so that they can be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts, including inorganic or organic acids, or in the case of the acidic form of the compound, salts may be prepared from inorganic or organic bases. In many cases, compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well known in the art, such as hydrochloric acid, sulfuric acid, hydrobromic acid, acetic acid, lactic acid, citric acid, or tartaric acid, which form acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, etc., which form base salts. Methods for preparing suitable salts are well established in the art.

[0102] Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethoxybenzoate ... Representative salts of the alkali or alkaline earth metals include sodium, lithium, potassium, calcium, and magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

[0103] The term "polypeptide" as used herein refers to a series of at least two amino acids linked together by a peptide bond. In some embodiments, a polypeptide can include at least 3-5 amino acids, each of which is linked to another amino acid by at least one peptide bond. One of skill in the art will understand that a polypeptide can include one or more "unnatural" amino acids, or other entities that can nevertheless be incorporated into a polypeptide chain. In some embodiments, a polypeptide can be glycosylated, e.g., a polypeptide can include one or more covalently attached sugar moieties. In some embodiments, a single "polypeptide" (e.g., an antibody polypeptide) can include two or more individual polypeptide chains, which can optionally be linked together, e.g., by one or more disulfide bonds or other means.

[0104] As used herein, the term "radioconjugate" refers to any complex that includes a radioisotope or radionuclide, such as any of the radioisotopes or radionuclides described herein.

[0105] As used herein, the term "radioimmunoconjugate" refers to any immunoconjugate that includes a radioisotope or radionuclide, such as any of the radioisotopes or radionuclides described herein.

[0106] The term "radioimmunotherapy" as used herein refers to a method of using a radioimmunoconjugate to produce a therapeutic effect. In some embodiments, radioimmunotherapy may include administering a radioimmunoconjugate to a subject in need thereof, where administration of the radioimmunoconjugate produces a therapeutic effect in the subject. In some embodiments, radioimmunotherapy may include administering a radioimmunoconjugate to a cell, where administration of the radioimmunoconjugate kills the cell. When radioimmunotherapy involves selective killing of a cell, in some embodiments, the cell is a cancer cell in a subject (e.g., a patient) with cancer.

[0107] As used herein, the term "radionuclide" refers to an atom capable of undergoing radioactive decay (e.g., 3 H, 14 C. 15 N, 18 F, 35 S, 47 Sc, 55 Co, 60 Cu, 61 Cu, 62 Cu, 64 Cu, 67 Cu, 75 Br, 76 Br, 77 Br, 89 Zr, 86 Y, 87 Y, 90 Y, 97 Ru, 99 Tc, 99m Tc 105 Rh, 109 Pd, 111 In, 123 I, 124 I, 125 I, 131 I, 149 Pm, 149 Tb, 153 Sm, 166 Ho, 177 Lu, 186 Re, 188 Re, 198 Au, 199 Au, 203 Pb, 211 At, 212 Pb, 212 Bi, 213 Bi, 223 Ra, 225 Ac, 227 Th, 229 Th, 66 Ga, 67 Ga, 68 Ga, 82 Rb, 117 mSn, 201Tl). The terms radionuclide, radioisotope, or radioisotope may also be used to describe radionuclides. As noted above, radionuclides can be used as detection agents. In some embodiments, the radionuclide is an alpha-emitting radionuclide.

[0108] By "subject" is meant a human (eg, a patient) or a non-human animal (eg, a mammal).

[0109] "Substantial identity" or "substantially identical" refers to a polypeptide sequence that has the same polypeptide sequence as a reference sequence, respectively, or has the same specified percentage of amino acid residues at corresponding positions in the reference sequence when the two sequences are optimally aligned. For example, an amino acid sequence that is "substantially identical" to a reference sequence has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the reference amino acid sequence. For polypeptides, the length of the comparison sequence will usually be at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 50, 75, 90, 100, 150, 200, 250, 300, or 350 consecutive amino acids (e.g., full-length sequence). Sequence identity may be measured using sequence analysis software, for example with default settings (e.g., the Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, WI 53705). Such software can match similar sequences by assigning degrees of homology to various substitutions, deletions, and other modifications.

[0110] The term "targeting moiety" as used herein refers to a molecule or any part of a molecule that binds to a given target.In some embodiments, targeting moiety is a protein or polypeptide, such as antibody or its antigen-binding fragment, nanobody, affibody, or the consensus sequence of fibronectin type III domain.In some embodiments, targeting moiety is a peptide or small molecule.

[0111] As used herein, the term "therapeutic moiety" refers to a molecule or any portion of a molecule that confers a therapeutic benefit. In some embodiments, the therapeutic moiety is a protein or polypeptide, such as an antibody, an antigen-binding fragment thereof. In some embodiments, the therapeutic moiety is a small molecule.

[0112] As used herein and as well understood in the art, "treating" a condition or "treatment" of a condition (e.g., a condition described herein, such as cancer) is an approach to obtain a beneficial or desired result, such as a clinical result. Beneficial or desired results may include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions; reduction in the severity of a disease, disorder, or condition; a stable (i.e., not worsening) state of a disease, disorder, or condition; prevention of the spread of a disease, disorder, or condition; delay or slowing of the progression of a disease, disorder, or condition; amelioration or alleviation of a disease, disorder, or condition; and remission (partial or complete), whether detectable or undetectable. In the context of cancer treatment, "ameliorating" may include, for example, a reduction in the incidence of metastasis, a reduction in tumor volume, a reduction in tumor vascularization, and / or a reduction in tumor growth rate. "Ameliorating" a disease, disorder, or condition means reducing the severity and / or undesirable clinical manifestations of the disease, disorder, or condition and / or slowing or prolonging the time course of progression compared to the severity or time course in the absence of treatment.

[0113] As used herein, the term "tumor-associated antigen" refers to an antigen that is present in significantly greater amounts on tumor cells than on normal cells.

[0114] As used herein, the term "tumor-specific antigen" refers to an antigen that is endogenously present only in tumor cells.

[0115] radioimmunoconjugate Radioimmunoconjugates suitable for use in accordance with the present disclosure generally have the following formula: AL 1 -XL 2 -ZB where each variable is as defined in the Summary section above. 225 Ac included 225 Ac] refers to radioimmunoconjugates.

[0116] In some embodiments, the radioimmunoconjugate has the following structure: [ka] where B is a targeting moiety (e.g., an antibody or antigen-binding fragment thereof, a peptide, or a small molecule).

[0117] In some embodiments, the radioimmunoconjugate has the following structure: [ka] where B is a targeting moiety (e.g., an antibody or antigen-binding fragment thereof, a peptide, or a small molecule). Antibodies and antigen-binding fragments thereof

[0118] Antibodies generally contain two identical light polypeptide chains and two identical heavy polypeptide chains linked together by disulfide bonds. The first domain located at the amino terminus of each chain is variable in amino acid sequence and provides the antibody binding specificity of the individual antibody. These are known as the variable heavy (VH) and variable light (VL) regions. The other domains of each chain are relatively invariant in amino acid sequence and are known as the constant heavy (CH) and constant light (CL) regions. Light chains generally contain one variable region (VL) and one constant region (CL). IgG heavy chains contain a variable region (VH), a first constant region (CH1), a hinge region, a second constant region (CH2), and a third constant region (CH3). In IgE and IgM antibodies, the heavy chain contains an additional constant region (CH4).

[0119] The antibodies described herein may include, for example, monoclonal antibodies, polyclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies, camelid antibodies, chimeric antibodies, single chain Fvs (scFvs), disulfide-linked Fvs (sdFvs), and anti-idiotypic (anti-Id) antibodies, and antigen-binding fragments of any of the above. In some embodiments, the antibodies or antigen-binding fragments thereof are humanized. In some embodiments, the antibodies or antigen-binding fragments thereof are chimeric. The antibodies may be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass.

[0120] The term "antigen-binding fragment" of an antibody, as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. Examples of binding fragments included in the term "antigen-binding fragment" of an antibody include Fab fragments, F(ab')2 fragments, Fd fragments, Fv fragments, scFv fragments, dAb fragments (Ward et al., (1989) Nature 341:544-546), and isolated complementarity determining regions (CDRs). In some embodiments, an "antigen-binding fragment" includes a heavy chain variable region and a light chain variable region. These antibody fragments can be obtained using conventional techniques known to those skilled in the art, and the fragments can be screened for utility in the same manner as intact antibodies.

[0121] The antibodies or fragments described herein can be produced by any method known in the art for the synthesis of antibodies (e.g., Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Brinkman et al., 1995, J. Immunol. Methods 182:41-50; WO 92 / 22324; WO 98 / 46645). Chimeric antibodies can be produced, for example, using the methods described in Morrison, 1985, Science 229:1202, and humanized antibodies can be produced, for example, by the methods described in U.S. Pat. No. 6,180,370.

[0122] Additional antibodies described herein are bispecific and multivalent antibodies, e.g., as described in Segal et al., J. Immunol. Methods 248:1-6 (2001); and Tutt et al., J. Immunol. 147:60 (1991).

[0123] In some embodiments, the antibody or antigen-binding fragment thereof is at least 100 kDa in size, eg, at least 150 kDa in size, at least 200 kDa in size, at least 250 kDa in size, or at least 300 kDa in size.

[0124] Insulin-like growth factor 1 (IGF-1R) antibodies Insulin-like growth factor 1 receptor is a transmembrane protein found on the surface of human cells that is activated by insulin-like growth factors 1 (IGF-1) and 2 (IGF-2). In some embodiments, the radioimmunoconjugate comprises an antibody against insulin-like growth factor-1 receptor (IGF-1R). Although not a typical oncogene, IGF-1R promotes cancer initiation and progression and plays a key role in mitogenic transformation and maintenance of the transformed phenotype. IGF-1R has been linked to the development of multiple common cancers, including breast cancer, lung cancer (e.g., non-small lung cancer), liver cancer, prostate cancer, pancreatic cancer, ovarian cancer, colon cancer, melanoma, adrenocortical carcinoma, and various types of sarcoma. IGF-1R signaling stimulates tumor cell proliferation and metabolism, supports angiogenesis, and confers protection from apoptosis. It influences metastatic factors (e.g., HIF-1-dependent hypoxia signaling), anchorage-independent growth, and growth and survival of tumor metastases after extravasation. IGF-1R is also involved in the generation, maintenance, and enrichment of therapy-resistant cancer stem cell populations.

[0125] Despite the wealth of data implicating the role of IGF-1R in cancer, therapeutics targeting IGF-1R have yet to demonstrate significant disease impact. There has been much speculation about this lack of efficacy, including failure to identify appropriate biomarkers for patient identification, the complexity and interdependence of the IGF-1 / IR signaling pathway, and the development of other growth hormone compensatory mechanisms [Beckwith and Yee, Mol Endocrinol, November 2015, 29(11):1549-1557]. However, radioimmunotherapy may provide a viable mechanism for treating cancers that overexpress the IGF-1 receptor by exploiting the ability of IGF-1R to undergo antibody-induced internalization and lysosomal degradation to deliver targeted radioisotopes within cancer cells. Internalization and lysosomal degradation of IGF-1R-targeted radioimmunoconjugates prolongs the residence time of the delivered radioisotope within cancer cells, thereby maximizing the chance of cell-death release. In the case of actinium-225, which produces four alpha particles per decay series, cell death can be achieved with as little as one atom of radionuclide delivered per cell [Sgouros et al., J Nucl Med. 2010, 51:311-2]. Cell death by direct DNA bombardment and destruction by alpha particles can occur in targeted cells or in a radius of two or three untargeted cells for a given alpha particle decay. In addition to having very high potential antitumor efficacy, IGF-1R-targeted radioimmunoconjugates may not develop mechanical resistance because they do not depend on blocking the ligand binding to the receptor to inhibit the oncological process, as is required for therapeutic antibodies.

[0126] Several IGF-1R antibodies have been developed and investigated for the treatment of various types of cancer, including figitumumab, cizutumumab, TAB-199, AVE1642 (also known as humanized EM164 and huEM164), BIIB002, lobatumumab, and teprotumumab. After binding to IGF-1R, these antibodies are internalized into the cell and degraded by lysosomal enzymes. The combination of overexpression and internalization in tumor cells offers the possibility of delivering detection agents directly to the tumor site while limiting the exposure of normal tissues to toxic agents.

[0127] In some embodiments, the light chain variable region of the IGF-1R antibody or antigen-binding fragment thereof comprises one, two or three complementarity determining regions (CDRs) CDR-L1, CDR-L2 and / or CDR-L3 having the amino acid sequence of AVE1642 as shown below, or a CDR region having an amino acid sequence that differs by one or two amino acids therefrom:

[0128] The CDRs of the light chain variable region of AVE1642 comprise the following sequence:

[0129] SEQ ID NO:1 (CDR-L1) RSSQSIVHSNVNTYLE Sequence number 2 (CDR-L2) KVSNRFS SEQ ID NO:3 (CDR-L3) FQGSHVPPT

[0130] In some embodiments, the light chain variable region of the IGF-1R antibody or antigen-binding fragment thereof comprises the light chain variable region of AVE1642 (SEQ ID NO:4), or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to the light chain variable region of AVE1642 (SEQ ID NO:4):

[0131] The light chain variable region of AVE1642 comprises the following sequence: SEQ ID NO:4 [ka]

[0132] In some embodiments, the heavy chain variable region of the IGF-1R antibody or antigen-binding fragment thereof comprises one, two or three complementarity determining regions (CDRs) CDR-H1, CDR-H2 and / or CDR-H3 having the amino acid sequence of AVE1642 as shown below, or a CDR region having an amino acid sequence that differs by one or two amino acids therefrom:

[0133] The CDRs of the heavy chain variable region of AVE1642 comprise the following sequences: SEQ ID NO:5 (CDR-H1) SYWMH SEQ ID NO:6 (CDR-H2) EINPSNGRTNYNQKFQG SEQ ID NO: 7 (CDR-H3) GRPDYYGSSKWYFDV

[0134] In some embodiments, the heavy chain variable region of the IGF-1R antibody or antigen-binding fragment thereof comprises the heavy chain variable region of AVE1642 (SEQ ID NO:8), or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to the heavy chain variable region of AVE1642 (SEQ ID NO:8):

[0135] The heavy chain variable region of AVE1642 comprises the following sequence: SEQ ID NO:8 [ka]

[0136] The light chain of AVE1642 contains the following sequence:

[0137] SEQ ID NO:22

[0138] [ka]

[0139]

[0140] The heavy chain of AVE1642 contains the following sequence:

[0141] SEQ ID NO:23

[0142] [ka] Endosialin (TEM-1) antibody

[0143] Endosialin (TEM-1) antibody Endosialin, also known as TEM-1 or CD-248, is an antigen expressed by tumor-associated endothelial cells, stromal cells, and pericytes.

[0144] Examples of endosialin antibodies include hMP-E-8.3 (disclosed in WO 2017 / 134234, the entire contents of which are incorporated herein by reference) and ontuxizumab (MORAb-004).

[0145] Fibroblast Growth Factor Receptor 3 (FGFR3) Antibodies Fibroblast growth factor receptor 3 (FGFR3) plays a key role during embryonic development, tissue homeostasis and metabolism by regulating a wide range of cellular processes, including proliferation, differentiation, migration and survival, in a context-dependent manner. It is overexpressed in many cancer types, often due to mutations that lead to constitutive activation.

[0146] In some embodiments, the methods provided include administering to a subject the therapeutic agent comprising an antibody or antigen-binding fragment thereof that targets FGFR3. 225 Ac] radioimmunoconjugate is used.

[0147] In certain embodiments, amino acid sequence variants of the antibody or antigen-binding fragment thereof are contemplated, such as variants capable of binding to human FGFR3 and / or mutant FGFR3 (e.g., mutant FGFR3 associated with cancer). For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody or antigen-binding fragment thereof. Amino acid sequence variants of the antibody or antigen-binding fragment thereof can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or antigen-binding fragment thereof, or by peptide synthesis. Such modifications include, for example, deletions and / or insertions and / or substitutions of residues within the amino acid sequence of the antibody or antigen-binding fragment thereof. Any combination of deletions, insertions and substitutions can be made to arrive at the final construct, as long as the final construct has the desired characteristics, e.g., antigen binding.

[0148] In some embodiments, the antibody or antigen-binding fragment thereof is an inhibitory antibody (also called an "antagonist antibody") or antigen-binding fragment thereof, e.g., the antibody or antigen-binding fragment thereof at least partially inhibits one or more functions of a target molecule (e.g., FGFR3), as further described herein.

[0149] Non-limiting examples of inhibitory antibodies include humanized monoclonal antibodies, such as MFGR1877S (CAS number 1312305-12-6; Genentech) (a human monoclonal antibody also known as bofatamab, the lyophilized form of which is also known as B-701 or R3Mab); PRO-001 (Prochon); PRO-007 (Fibron); IMC-D11 (Imclone); and AV-370 (Aveo Pharmaceuticals). (See, e.g., U.S. Pat. No. 8,410,250; U.S. Patent Application Publication No. 10,208,120; WO 2002102972 A2, WO 2002102973 A2, WO 2007144893 A2, WO 2010002862 A2, and WO 2010048026 A2.)

[0150] In some embodiments, the antibody or antigen-binding fragment thereof is an agonist antibody (also known as a stimulatory antibody).

[0151] In some embodiments, the antibody or antigen-binding fragment thereof is neither an agonist nor an antagonist, or is not characterized as an agonist or antagonist.

[0152] Further known FGFR3 antibodies include, for example, mouse monoclonal antibodies such as 1G6, 6G1 and 15B2 from Genentech (see, for example, U.S. Patent No. 8,410,250), B9 (Sc-13121) (Santa Cruz Biotechnology), MAB766 (clone 136334) (R&D systems), MAB7661 (clone 136318) (R&D systems) and OTI1B10 (OriGene); rabbit polyclonal antibodies such as ab10651 (Abcam); rabbit monoclonal antibodies such as C51F2 (catalog number 4574) (Cell Signaling Technology).

[0153] In certain embodiments of the present disclosure, the antibody or antigen-binding fragment thereof comprises the specific heavy chain complementarity determining regions CDR-H1, CDR-H2 and / or CDR-H3 described herein. In some embodiments, the complementarity determining regions (CDRs) of the antibody or antigen-binding fragment thereof are adjacent to framework regions. The heavy or light chain of the antibody or antigen-binding fragment thereof that contains three CDRs typically contains four framework regions.

[0154] In some embodiments, the heavy chain variable region of the FGFR3 antibody or antibody-binding fragment thereof comprises one, two or three complementarity determining regions (CDRs) CDR-H1, CDR-H2 and / or CDR-H3 having the amino acid sequences as set forth below, or CDR regions having amino acid sequences that differ by one or two amino acids therefrom: CDR-H1: GFTFTSTGIS (SEQ ID NO: 9) CDR-H2: GRIYPTSGSTNYADSV (SEQ ID NO: 10) CDR-H3: TYGIYDLYVDYTEYVMDY (SEQ ID NO: 11) or ARTYGIYDLYVDYTEYVMDY (SEQ ID NO: 12)

[0155] In some embodiments, the light chain variable region of the FGFR3 antibody or antigen-binding fragment thereof comprises one, two or three complementarity determining regions (CDRs) CDR-L1, CDR-L2 and / or CDR-L3 having the amino acid sequences as set forth below, or CDR regions having amino acid sequences that differ by one or two amino acids therefrom: CDR-L1: RASQDVDTSLA (SEQ ID NO: 13) CDR-L2: SASFLYS (SEQ ID NO: 14) CDR-L3: QQSTGHPQT (SEQ ID NO: 15)

[0156] In some embodiments, the antibody or antigen-binding fragment thereof has CDR sequences having the amino acid sequences of SEQ ID NOs: 9, 10, 11, 13, 14, and 15 without any mutations. For example, in some embodiments, the antibody or antigen-binding fragment thereof comprises heavy chain complementarity determining regions CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 9, 10, and 11, and heavy chain complementarity determining regions CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NOs: 13, 14, and 15.

[0157] In some embodiments, the antibody or antigen-binding fragment thereof has CDR sequences having the amino acid sequences of SEQ ID NOs: 9, 10, 12, 13, 14, and 15 without any mutations. For example, in some embodiments, the antibody or antigen-binding fragment thereof comprises heavy chain complementarity determining regions CDR-H1, CDR-H2, and CDR-H3 having the amino acid sequences of SEQ ID NOs: 9, 10, and 12, and heavy chain complementarity determining regions CDR-L1, CDR-L2, and CDR-L3 having the amino acid sequences of SEQ ID NOs: 13, 14, and 15.

[0158] In some embodiments, the heavy chain variable region of the FGFR3 antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:16, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:16. [ka]

[0159] In some embodiments, the heavy chain variable region of the FGFR3 antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:18, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:18. [ka]

[0160] In some embodiments, the heavy chain of the FGFR3 antibody comprises a constant region having an amino acid sequence of SEQ ID NO:20, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:20. [ka] [ka]

[0161] In some embodiments, the heavy chain of the FGFR3 antibody comprises an amino acid sequence of SEQ ID NO:24, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:24. [ka]

[0162] In some embodiments, the light chain variable region of the FGFR3 antibody or antigen-binding fragment thereof comprises an amino acid sequence of SEQ ID NO: 17, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO: 17. [ka]

[0163] In some embodiments, the light chain variable region of the FGFR3 antibody or antigen-binding fragment thereof comprises the amino acid sequence of SEQ ID NO:19, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:19. [ka]

[0164] In some embodiments, the light chain of the FGFR3 antibody comprises a constant region having an amino acid sequence of SEQ ID NO:21, or an amino acid sequence that differs therefrom by 1, 2, 3, or 4 amino acids, or an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% identical to SEQ ID NO:21. [ka]

[0165] In some embodiments, the FGFR3 antibody or antigen-binding fragment thereof comprises at least one, two, three, four, five, or six complementarity determining regions (CDRs) selected from the group consisting of: CDR-H1 comprising the amino acid sequence of SEQ ID NO:9 or an amino acid sequence which differs therefrom by 1 or 2 amino acids; CDR-H2 comprising the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence which differs therefrom by 1 or 2 amino acids; CDR-H3 comprising the amino acid sequence of SEQ ID NO: 11 or 12, or an amino acid sequence which differs by 1 or 2 amino acids from SEQ ID NO: 11 or 12; CDR-L1 comprising the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence which differs therefrom by 1 or 2 amino acids; CDR-L2 comprising the amino acid sequence of SEQ ID NO: 14, or an amino acid sequence which differs therefrom by 1 or 2 amino acids; and A CDR-L3 comprising the amino acid sequence of SEQ ID NO: 15, or an amino acid sequence which differs therefrom by 1 or 2 amino acids.

[0166] In some embodiments, the antibody or antigen-binding fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:18, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:17 or SEQ ID NO:19.

[0167] In some embodiments, the antibody or antigen-binding fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:16, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:17.

[0168] In some embodiments, the antibody or antigen-binding fragment thereof comprises (i) a heavy chain variable domain comprising the amino acid sequence of SEQ ID NO:18, and (ii) a light chain variable domain comprising the amino acid sequence of SEQ ID NO:19.

[0169] In some embodiments, the FGFR3 antibody is MFGR1877S (bofatamab).

[0170] Nanobody Nanobodies are antibody fragments consisting of a single monomeric variable antibody domain. Nanobodies are sometimes called single domain antibodies. Like antibodies, nanobodies selectively bind to a specific antigen. Nanobodies can be heavy chain variable domains or light chain domains. Nanobodies can be naturally occurring or bioengineered products. Nanobodies can be biologically engineered by site-directed mutagenesis or mutagenic screening (e.g., phage display, yeast display, bacterial display, mRNA display, ribosome display).

[0171] Affibody Affibodies are polypeptides or proteins engineered to bind to a specific antigen. Thus, affibodies can be thought of as mimicking certain functions of antibodies. Affibodies can be engineered variants of the B domain within the immunoglobulin binding region of Staphylococcus protein A. Affibodies can be engineered variants of the Z domain, the B domain with low affinity to the Fab region. Affibodies can be biologically engineered by site-directed mutagenesis or mutagenic screening (e.g., phage display, yeast display, bacterial display, mRNA display, ribosome display).

[0172] Affibody molecules have been generated that exhibit specific binding to a variety of different proteins (e.g., insulin, fibrinogen, transferrin, tumor necrosis factor-α, IL-8, gp120, CD28, human serum albumin, IgA, IgE, IgM, HER2, and EGFR) with affinities (K ) in the μM to pM range. d ) is shown.

[0173] Fibronectin type III domain Fibronectin type III domains are evolutionarily conserved protein domains found in diverse extracellular proteins. Fibronectin type III domains have been used as molecular scaffolds to generate molecules that can selectively bind to specific antigens. Fibronectin type III domain (FN3) variants engineered for selective binding are sometimes called monobodies. FN3 domains may be biologically engineered by site-directed mutagenesis or mutagenic screening (e.g., CIS-display, phage display, yeast display, bacterial display, mRNA display, ribosome display).

[0174] Modified Polypeptides The polypeptide used according to the present disclosure may have a modified amino acid sequence. The modified polypeptide may be substantially identical to the corresponding reference polypeptide (e.g., the amino acid sequence of the modified polypeptide may have at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence of the reference polypeptide). In certain embodiments, the modification does not significantly destroy the desired biological activity (e.g., binding to IGF-1R or endosialin). Modifications may reduce, have no effect on, or increase (e.g., at least 5%, 10%, 20%, 25%, 35%, 50%, 60%, 70%, 75%, 80%, 90%, or 95%) the biological activity of the original polypeptide. Modified polypeptides may possess or optimize polypeptide characteristics such as in vivo stability, bioavailability, toxicity, immunological activity, immunological identity, and conjugation properties.

[0175] Modifications include those that result from natural processes such as post-translational processing, or those that result from chemical modification techniques known in the art. Modifications can occur anywhere in a polypeptide, including the polypeptide backbone, the amino acid side chains, and the amino or carboxy termini. The same type of modification can be present in the same or different degrees at several sites in a given polypeptide, and a polypeptide can contain more than one type of modification. Polypeptides can be branched as a result of ubiquitination, and can be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides can result from post-translational natural processes or can be synthetically produced. Other modifications include pegylation, acetylation, acylation, addition of acetomidomethyl (Acm) groups, ADP-ribosylation, alkylation, amidation, biotinylation, carbamoylation, carboxyethylation, esterification, covalent attachment to a flavin, covalent attachment to a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a drug, covalent attachment of a marker (e.g., fluorescent or radioactive), covalent attachment of a lipid or lipid derivative, phosphatidylinositol, or the like. These include covalent attachment of toll, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cystine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins, such as arginylation and ubiquitination.

[0176] Modified polypeptides may also include insertions, deletions, or conservative or non-conservative substitutions (e.g., D-amino acids, desamino acids) of amino acids in the polypeptide sequence (e.g., if such changes do not substantially alter the biological activity of the polypeptide). In particular, the addition of one or more cysteine ​​residues to the amino or carboxy termini of the polypeptides can facilitate conjugation of these polypeptides, for example, by disulfide bonds. For example, the polypeptides can be modified to include a single cysteine ​​residue at the amino terminus or a single cysteine ​​residue at the carboxy terminus. Amino acid substitutions can be conservative (i.e., a residue is replaced by another of the same general type or group) or non-conservative (i.e., a residue is replaced by an amino acid of another type). Additionally, naturally occurring amino acids can be used in place of non-naturally occurring amino acids (i.e., non-naturally occurring conservative amino acid substitutions or non-naturally occurring non-conservative amino acid substitutions).

[0177] Synthetically produced polypeptides can include substitutions of amino acids that are not naturally encoded in DNA (e.g., non-naturally occurring or unnatural amino acids). Examples of non-naturally occurring amino acids include D-amino acids, N-protected amino acids, amino acids having an acetylaminomethyl group attached to the sulfur atom of cysteine, pegylated amino acids, amino acids with the formula NH2(CH2) n COOH omega amino acids (where n is 2-6), neutral non-polar amino acids such as sarcosine, t-butylalanine, t-butylglycine, n-methylisoleucine, and norleucine. Phenylglycine can replace Trp, Tyr, or Phe. Citrulline and methionine sulfoxide are neutral non-polar, cysteic acid is acidic, and ornithine is basic. Proline can be replaced with hydroxyproline and retain the properties that give it a conformation.

[0178] Analogs may be generated by substitution mutagenesis and may retain the biological activity of the original polypeptide. Examples of substitutions identified as "conservative substitutions" are shown in Table 1. If such substitutions result in undesirable changes, other types of substitutions referred to as "exemplary substitutions" in Table 1 or further described herein in relation to amino acid classes are introduced and the products screened. Table 1: Amino acid substitutions [Table 1]

[0179] Substantial modification of function or immunological identity is achieved by selecting substitutions that differ significantly in their effect on (a) maintaining the structure of the polypeptide backbone in the area of ​​substitution, e.g., as a sheet or helical conformation, (b) maintaining the charge or hydrophobicity of the molecule at the target site, or (c) maintaining side-chain bulk.

[0180] Chelate part Examples of suitable chelating moieties include DOTA (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetraacetic acid), DOTMA (1R,4R,7R,10R)-α,α',α”,α”'-tetramethyl-1,4,7,10 tetraazacyclododecane-1,4,7,10 tetraacetic acid, DOTAM (1,4,7,10 tetrakis(carbamoylmethyl)-1,4,7,10 tetraazacyclododecane), DOTPA (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetra propionic acid), DO3AM-acetic acid (2-(4,7,10 tris(2-amino-2-oxoethyl)-1,4,7,10 tetraazacyclododecan-1-yl)acetic acid), DOTP (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetra(methylenephosphonic acid)), DOTA-4AMP (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetrakis(acetamido-methylenephosphonic acid), CB-TE2A (1,4,8,11 tetraazabicyclo[6.6.2]hexadecane-4,11 diacetic acid), NOTA (1,4,7 triazacyclononane-1,4,7 triacetic acid), NOTP (1,4,7 triazacyclononane-1,4,7 tri(methylenephosphonic acid), TETPA (1,4,8,11 tetraazacyclotetradecane-1,4,8,11 tetrapropionic acid), TETA (1,4,8,11 tetraazacyclotetradecane-1,4,8,11 tetraacetic acid), HEHA (1,4,7,10,13,16 hexaazacyclohexadecane-1,4,7,10,13,16 hexaacetic acid), PEPA (1,4,7,10,13 pentaazacyclopentadecane-N,N',N",N"',N""-pentaacetic acid), H4octapa (N,N'-bis(6-carboxy-2-pyridylmethyl)-ethylenediamine-N,N'-diacetic acid), H2dedpa (1 ,2-[[6-(carboxy)-pyridin-2-yl]-methylamino]ethane), H6phospa (N,N'-(methylenephosphonate)-N,N'-[6-(methoxycarbonyl)pyridin-2-yl]-methyl-1,2-diaminoethane), TTHA (triethylenetetramine-N,N,N',N",N"',N"'-hexaacetic acid), DO2P (tetraazacyclododecane dimethanephosphonic acid), HP-DO3A (hydroxypropyltetraazacyclododecane triacetic acid), EDTA (ethylenediaminetetraacetic acid), deferoxamine, DTPA (diethylenetriaminepentaacetic acid), DTPA-BMA (diethylenetriaminepentaacetic acid-bismethylamide), HOPO (octadentate hydroxypyridinone), or porphyrin.

[0181] Preferably, the chelating moiety is DOTA (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetraacetic acid), DOTMA (1R,4R,7R,10R)-α,α',α",α"'-tetramethyl-1,4,7,10 tetraazacyclododecane-1,4,7,10 tetraacetic acid, DOTAM (1,4,7,10 tetrakis(carbamoylmethyl)-1,4,7,10 tetraazacyclododecane), DO3AM-acetic acid (2-(4,7,10 tris(2-amino-2-oxoethyl)-1,4,7,10 tetraacetate), The amide is selected from 1,4,7,10 tetrakis(acetamido-methylene phosphonic acid), DOTP (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetra(methylene phosphonic acid), DOTA-4AMP (1,4,7,10 tetraazacyclododecane-1,4,7,10 tetrakis(acetamido-methylene phosphonic acid), NOTA (1,4,7 triazacyclononane-1,4,7 triacetic acid), and HP-DO3A (10-(2-hydroxypropyl)-1,4,7 tetraazacyclododecane-1,4,7 triacetic acid).

[0182] In some embodiments, the chelating moiety is DOTA.

[0183] In some embodiments, the chelating moieties are useful as detection agents and thus radioimmunoconjugates containing such detectable chelating moieties can be used as diagnostic or theranostic agents.

[0184] Linker [ 225 With reference to the [Ac]-radioimmunoconjugate, the linker has the following formula: AL 1 -XL 2 -ZB and the linker is typically -L 1 -XL 2 -Z-, wherein L 1 is a bond or optionally substituted C 1~6 Alkyl or C 1~6 is heteroalkyl, X is -C(O)NR1 -*, -NR 1 C(O)-*, -OC(O)NR 1 -*, -NR 1 C(O)O-*, -NR 1 C(O)NR 1 -, -CH2-Ph-C(O)NR 1 -*, -NR 1 C(O)-Ph-CH2-*, -O-, or -NR 1 -, where "*" represents L 2 Each R 1 are independently hydrogen or C 1~6 is alkyl, L 2 is replaced by C 1~50 Alkyl or C 1~50 is heteroalkyl, Z is -C(O)-, -CH2-, -OC(O)-#, -C(O)O-#, -NR 2 C(O)-#, -C(O)NR 2 -#, or -NR 2 -, where "#" indicates the point of attachment to B, and each R 2 are independently hydrogen or C 1~6 It is an alkyl.

[0185] In some embodiments, L 1 is replaced by C 1~6 For example, L 1 is -CH2CH2-. For example, L 1 The structure: [ka] wherein R 2 is hydrogen or -COH.

[0186] In some embodiments, X is -C(O)NR 1 -*, where "*" is L 2 indicates the point of attachment to R 1 is H.

[0187] In some embodiments, L2 is replaced by C 1~50 Alkyl (e.g., C 1~40 Alkyl, C 1~30 Alkyl, C 1~20 Alkyl, C 2~18 Alkyl, C 3~16 Alkyl, C 4~14 Alkyl, C 5~12 Alkyl, C 6~10 Alkyl, C 8~10 Alkyl or C 10 For example, L 2 is C as shown below. 10 Alkyl: [ka] .

[0188] In some embodiments, L 2 is replaced by C 1~50 Heteroalkyl (e.g., C 1~40 Heteroalkyl, C 1~30 Heteroalkyl, C 1~20 Heteroalkyl, C 2~18 Heteroalkyl, C 3~16 Heteroalkyl, C 4~14 Heteroalkyl, C 5~12 Heteroalkyl, C 6~10 Heteroalkyl, C 8~10 Heteroalkyl, C4 heteroalkyl, C6 heteroalkyl, C8 heteroalkyl, C 10 Heteroalkyl, C 12 Heteroalkyl, C 16 Heteroalkyl, C 20 Heteroalkyl or C 24 In certain embodiments, L is 2is an optionally substituted C olefin comprising a polyethylene glycol (PEG) moiety containing 1 to 20 oxyethylene (-O-CH2-CH2-) units, e.g., 2 oxyethylene units (PEG2), 3 oxyethylene units (PEG3), 4 oxyethylene units (PEG4), 5 oxyethylene units (PEG5), 6 oxyethylene units (PEG6), 7 oxyethylene units (PEG7), 8 oxyethylene units (PEG8), 9 oxyethylene units (PEG9), 10 oxyethylene units (PEG10), 12 oxyethylene units (PEG12), 14 oxyethylene units (PEG14), 16 oxyethylene units (PEG16) or 18 oxyethylene units (PEG18). 1~50 It is heteroalkyl.

[0189] In certain embodiments, L 2 is an optionally substituted C 2 -containing polyethylene glycol (PEG) moiety containing 1 to 20 oxyethylene (-O-CH2-CH2-) units or portions thereof. 1~50 Heteroalkyl. For example, L 2 is a linker containing PEG3 as shown below: [ka] .

[0190] In some embodiments, Z is -C(O)- or -CH2-. In some embodiments, Z is -C(O)- and is the point of conjugation to B via a lysine residue from B.

[0191] In certain embodiments, the formula AL 1 -XL 2 -ZB has the following structure: [ka] (In the formula, Y 1 is -CH2OCH2L 2 -B, C(O)L 2 -B, or C(S)L 2-B and Y 2 is -CH2CO2H, or Y 1 is H and Y 2 L 1 -XL 2 -B).

[0192] Checkpoint inhibitors In some embodiments, the checkpoint inhibitor is co-administered with the radioimmunoconjugate. In general, suitable checkpoint inhibitors inhibit immunosuppressive checkpoint proteins. In some embodiments, the checkpoint inhibitor inhibits a protein selected from the group consisting of cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), programmed cell death 1 (PD-1), programmed cell death ligand-1 (PD-L1), LAG-3, T-cell immunoglobulin mucin 3 (TIM-3), and killer immunoglobulin-like receptor (KIR).

[0193] For example, in some embodiments, the checkpoint inhibitor can bind to CTLA-4, PD-1, or PD-L1. In some embodiments, the checkpoint inhibitor disrupts the interaction (e.g., disrupts the binding) of PD-1 and PD-L1.

[0194] In some embodiments, the checkpoint inhibitor is a small molecule.

[0195] In some embodiments, the checkpoint inhibitor is an antibody or an antigen-binding fragment thereof, such as a monoclonal antibody. In some embodiments, the checkpoint inhibitor is a human or humanized antibody or an antigen-binding fragment thereof. In some embodiments, the checkpoint inhibitor is a murine antibody or an antigen-binding fragment thereof.

[0196] In some embodiments, checkpoint inhibitor is CTLA-4 antibody.Non-limiting examples of CTLA-4 antibody include BMS-986218, BMS-986249, ipilimumab, tremelimumab (formerly ticilimumab, CP-675,206), MK-1308 and REGN-4659.Another example of CTLA-4 antibody is 4F10-11, which is a mouse monoclonal antibody.

[0197] In some embodiments, the checkpoint inhibitor is a PD-1 antibody. Non-limiting examples of PD-1 antibodies include camrelizumab, cemiplumab, nivolumab, pembrolizumab, sintilimab, tislelizumab and toripalimab. A further example of a PD-1 antibody is the mouse monoclonal antibody RMP1-14. In some embodiments, the checkpoint inhibitor is pembrolizumab.

[0198] In some embodiments, the checkpoint inhibitor is a PD-L1 antibody. Non-limiting examples of PD-L1 antibodies include atezolizumab, avelumab, and durvalumab.

[0199] In some embodiments, a combination of more than one checkpoint inhibitor is used, for example, in some embodiments, both a CTLA-4 inhibitor and a PD-1 or PD-L1 inhibitor are used.

[0200] Combination therapy As provided above, the present disclosure provides a method for the effective treatment of cancer at specific dose levels, 225 The present invention relates to a combination therapy comprising a β-radioimmunoconjugate and one or more checkpoint inhibitors.

[0201] In some embodiments, the combination therapy includes a PD-1 inhibitor or a CTLA-4 inhibitor chelated with one of the following compounds: 225 Ac, [ 225 Ac]-radioimmunoconjugates, including: [ka] , B is an IGF-1R antibody or antigen-binding domain thereof; [ka] , B is an FGFR3 antibody or an antigen-binding domain thereof; [ka] , B is the TEM-1 antibody or an antigen-binding domain thereof; [ka] , B is an IGF-1R antibody, an FGFR3 antibody or a TEM-1 antibody, or an antigen-binding domain thereof; [ka] , B is an IGF-1R antibody, an FGFR3 antibody or a TEM-1 antibody, or an antigen-binding domain thereof; [ka] , B is an IGF-1R antibody, an FGFR3 antibody or a TEM-1 antibody, or an antigen-binding domain thereof; [ka] , B is an IGF-1R antibody, an FGFR3 antibody or a TEM-1 antibody, or an antigen-binding domain thereof.

[0202] In some embodiments, the combination therapy comprises a PD-1 inhibitor and one of the following compounds: [ka] , (B is TAB-199 or AVE1642) 225 Ac included 225 Ac]-radioimmunoconjugate.

[0203] In some embodiments, the combination therapy comprises pembrolizumab and one of the following compounds: [ka] , (B is AVE1642) Chelated with 225 Ac included 225 Ac]-radioimmunoconjugate.

[0204] Subject In some disclosed methods, a therapy (e.g., including a therapeutic agent) is administered to a subject. In some embodiments, the subject is a mammal, e.g., a human.

[0205] In some embodiments, the subject has been administered or is currently being administered another treatment. For example, in some embodiments, the subject has been administered or is currently being administered a radioimmunoconjugate. In some embodiments, the subject has been administered or is currently being administered a checkpoint inhibitor.

[0206] In some embodiments, the subject has cancer or is at risk of developing cancer. For example, the subject has been diagnosed with cancer. The cancer may be primary or metastatic. The subject may have cancer at any stage, such as stage I, stage II, stage III, or stage IV, with or without lymph node involvement and with or without metastasis. The provided compositions can prevent or reduce further growth of the cancer and / or otherwise reverse the cancer (e.g., prevent or reduce metastasis). In some embodiments, the subject does not have cancer, but has been determined to be at risk of developing cancer due to the presence of one or more risk factors, such as, for example, environmental exposure, the presence of one or more genetic mutations or variants, family history, etc. In some embodiments, the subject has not been diagnosed with cancer.

[0207] In some embodiments, the cancer is a solid tumor.

[0208] In some embodiments, the solid tumor cancer is breast cancer (e.g., TNBC), non-small cell lung cancer, small cell lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, colorectal cancer, cervical cancer, endometrial cancer, sarcoma, adrenocortical carcinoma, neuroendocrine carcinoma, Ewing's sarcoma, multiple myeloma, or acute myeloid leukemia.

[0209] In some embodiments, the cancer is a non-solid (e.g., liquid (e.g., blood)) cancer.

[0210] Administration and Dosage Effective doses and lower effective doses The present disclosure provides combination therapy, in which the amount of each therapeutic agent may or may not be therapeutically effective by itself.For example, a method is provided that includes administering a first treatment and a second treatment together in an amount effective for treating or improving a disorder, for example, cancer.In some embodiments, at least one of the first and second treatments is administered to the subject at a lower effective dose.In some embodiments, both the first and second treatments are administered at a lower effective dose.

[0211] In some embodiments, the first treatment comprises a radioimmunoconjugate and the second treatment comprises a checkpoint inhibitor.

[0212] In some embodiments, the first treatment comprises a checkpoint inhibitor and the second treatment comprises a radioimmunoconjugate.

[0213] In some embodiments, the therapeutic combinations disclosed herein are administered to a subject in a manner (e.g., dosage and timing) sufficient to cure or at least partially arrest the symptoms of the disorder and its complications. In the context of a single treatment ("monotherapy"), an amount sufficient to achieve this purpose is defined as a "therapeutically effective amount", i.e., an amount of compound sufficient to substantially improve at least one symptom or medical condition associated with the disease. A "therapeutically effective amount" usually varies depending on the therapeutic agent. For known therapeutic agents, the relevant therapeutically effective amount is known or can be easily determined by those skilled in the art.

[0214] For example, in the treatment of cancer, a drug or compound that reduces, prevents, delays, inhibits, or suppresses any symptoms of a disease or condition is therapeutically effective.A therapeutically effective amount of a drug or compound is not necessary to cure a disease or condition, but provides a treatment for a disease or condition such that the onset of the disease or condition is delayed, hindered, or prevented, or the symptoms of the disease or condition are improved, or the duration of the disease or condition is changed, or for example, the disease or condition becomes less severe in an individual or the recovery is accelerated.For example, a treatment can be therapeutically effective if it causes cancer to regress or the growth of cancer to slow down.

[0215] Effective dosing regimens for these uses (e.g., amounts of each therapeutic agent, relative timing of treatment, etc.) may depend on the severity of the disease or condition, as well as the weight and general condition of the subject. For example, the therapeutically effective amount of a particular composition containing a therapeutic agent to be applied to a mammal (e.g., a human) may be determined by one of skill in the art, taking into account individual differences in the age, weight, and condition of the mammal. Because certain conjugates of the present disclosure have an enhanced ability to target and remain in cancer cells, the dosage of these compounds may be lower than the equivalent dose required for therapeutic effect of the unconjugated agent (e.g., less than or equal to about 90%, 75%, 50%, 40%, 30%, 20%, 15%, 12%, 10%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%). Therapeutically effective and / or optimal amounts may also be empirically determined by one of skill in the art. Thus, lower effective doses may also be determined by one of skill in the art.

[0216] In order to carry out the method of the present invention, 225 Ac]-radioimmunoconjugates are typically administered at a dose of about 10 kBq to about 400 kBq / kg. 225Ac]-radioimmunoconjugate is administered at a dose of 10 kBq to about 200 kBq per kg of the patient's body weight (e.g., about 10 kBq to about 150 kBq / kg, about 10 kBq to about 120 kBq / kg, about 10 kBq to about 100 kBq / kg; about 20 kBq to about 150 kBq / kg, about 20 kBq to about 120 kBq / kg, about 20 kBq to about 10 0 kBq / kg; about 30 kBq to about 150 kBq / kg, about 30 kBq to about 120 kBq / kg, about 30 kBq to about 100 kBq / kg; about 40 kBq to about 150 kBq / kg, about 40 kBq to about 120 kBq / kg, about 40 kBq to about 100 kBq / kg, or about 40 kBq to about 80 kBq / kg).

[0217] In some embodiments, [ 225 Ac]-radioimmunoconjugate is administered at a dose of about 30 kBq to about 120 kBq per kg of body weight of the patient (e.g., about 35 kBq / kg, about 40 kBq / kg, about 45 kBq / kg, about 50 kBq / kg, about 55 kBq / kg, about 60 kBq / kg, about 65 kBq / kg, about 70 kBq / kg, about 75 kBq / kg, about 80 kBq / kg, about 85 kBq / kg, about 90 kBq / kg, about 95 kBq / kg, about 100 kBq / kg, about 105 kBq / kg, about 110 kBq / kg, or about 115 kBq / kg).

[0218] In some embodiments, [ 225 Ac]-radioimmunoconjugate is administered to the patient as a unit dose of about 1-30 MBq (e.g., about 1-25 MBq, about 1-20 MBq, about 1-15 MBq, about 1-10 MBq; about 2-25 MBq, about 2-20 MBq, about 2-15 MBq, about 2-10 MBq; about 3-25 MBq, about 3-20 MBq, about 3-15 MBq, about 3-10 MBq; about 5-25 MBq, about 5-20 MBq, about 5-15 MBq, about 5-10 MBq).

[0219] In some embodiments, [ 225Ac] radioimmunoconjugate is administered to the patient as a unit dose of about 5 to 15 MBq (e.g., about 6 MBq, about 7 MBq, about 8 MBq, about 9 MBq, about 10 MBq, about 11 MBq, about 12 MBq, about 13 MBq, or about 14 MBq).

[0220] In some embodiments, [ 225 Ac] radioimmunoconjugate is administered to the patient as a unit dose of about 20 to 30 MBq (e.g., about 21 MBq, about 22 MBq, about 23 MBq, about 24 MBq, about 25 MBq, about 26 MBq, about 27 MBq, about 28 MBq, or about 29 MBq).

[0221] Single or multiple administrations of the composition (e.g., a pharmaceutical composition containing a therapeutic agent) can be carried out with the dose level and pattern selected by the treating physician. The dose and administration schedule can be determined and adjusted based on the severity of the subject's disease or condition, and these may be monitored throughout the course of treatment according to methods commonly practiced by clinicians or methods described herein.

[0222] In some embodiments, the unit dose can be administered to a subject (e.g., a patient) twice a day, three times a day, or four times a day. 225 When the [Ac]-radioimmunoconjugate is administered as a unit dose of about 10-30 MBq, it can be administered to a patient twice daily for a total dose of about 20 MBq to about 60 MBq.

[0223] In the disclosed combination therapy method, the first and second treatments can be administered to the subject sequentially or simultaneously.For example, the first composition comprising the first therapeutic agent and the second composition comprising the second therapeutic agent can be administered to the subject sequentially or simultaneously.Alternatively, or in addition, the composition comprising the combination of the first therapeutic agent and the second therapeutic agent can be administered to the subject.

[0224] In some embodiments, the radioimmunoconjugate is administered in a single dose. In some embodiments, the radioimmunoconjugate is administered multiple times. When the radioimmunoconjugate is administered multiple times, the dose of each administration may be the same or different.

[0225] In some embodiments, the checkpoint inhibitor is administered in a single dose. In some embodiments, the checkpoint inhibitor is administered multiple times (e.g., at least twice, at least three times, etc.). In some embodiments, the checkpoint inhibitor is administered multiple times according to a regular or semi-regular schedule, for example, about once every two weeks, once a week, twice a week, three times a week, or more than three times a week. When the checkpoint inhibitor is administered multiple times, the dose of each administration can be the same or different. For example, the checkpoint inhibitor can be administered in an initial dose, and the dosage of the checkpoint inhibitor thereafter can be higher or lower than the initial dose.

[0226] In some embodiments, the first dose of the checkpoint inhibitor is administered simultaneously with the first dose of the radioimmunoconjugate. In some embodiments, the first dose of the checkpoint inhibitor is administered before the first dose of the radioimmunoconjugate. In some embodiments, the first dose of the checkpoint inhibitor is administered after the first dose of the radioimmunoconjugate. In some embodiments, a subsequent dose of the checkpoint inhibitor is administered.

[0227] In some embodiments, the radioimmunoconjugate (or composition thereof) and the checkpoint inhibitor (or composition thereof) are administered within 28 days (e.g., within 14, 7, 6, 5, 4, 3, 2, or 1 day) of each other.

[0228] In some embodiments, the radioimmunoconjugate (or composition thereof) and the checkpoint inhibitor (or composition thereof) are administered within 90 days (e.g., within 80, 70, 60, 50, 40, 30, 20, 10, 5, 4, 3, 2, or 1 day) of each other. In various embodiments, the checkpoint inhibitor is administered simultaneously with the radioimmunoconjugate. In various embodiments, the checkpoint inhibitor is administered multiple times following a first administration of the radioimmunoconjugate.

[0229] In some embodiments, the composition (e.g., a composition comprising a radioimmunoconjugate) is administered for radiation treatment planning or diagnostic purposes. When administered for radiation treatment planning or diagnostic purposes, the composition may be administered to the subject in an amount effective to determine a diagnostically effective dose and / or a therapeutically effective dose. In some embodiments, a first dose of the disclosed conjugate or a composition thereof (e.g., a pharmaceutical composition) is administered in an amount effective for a radiation treatment planning, followed by administration of a combination therapy comprising a conjugate disclosed herein and another therapeutic agent.

[0230] Pharmaceutical compositions containing one or more agents (e.g., radioimmunoconjugates and / or checkpoint inhibitors) can be formulated for use according to the disclosed methods and systems in a variety of drug delivery systems. One or more physiologically acceptable excipients or carriers can also be included in the composition for suitable formulation. Examples of suitable formulations can be found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, 17th ed., 1985. For a brief overview of methods for drug delivery, see, for example, Langer (Science 249:1527-1533, 1990).

[0231] formulation The pharmaceutical composition may be formulated for local administration, such as by parenteral, intranasal, topical, oral, or transdermal means, for prophylactic and / or therapeutic treatment. The pharmaceutical composition may be administered parenterally (e.g., by intravenous, intramuscular, or subcutaneous injection), or by oral ingestion, or by local application or intraarticular injection to the area affected by vascular or cancerous conditions. Additional exemplary routes of administration include intravascular, intraarterial, intratumoral, intraperitoneal, intraventricular, intraepithelial, as well as nasal, ocular, intrascleral, intraorbital, rectal, topical, or aerosol inhalation administration. Also specifically contemplated are sustained release administration, such as by means of depot injection or erodible implants or components. Suitable compositions include compositions comprising an agent (e.g., a compound disclosed herein) dissolved or suspended in an acceptable carrier, preferably an aqueous carrier, such as water, buffered water, saline, or PBS, among others, for parenteral administration. The composition may contain pharma- ceutically acceptable auxiliary substances, such as pH adjusting and buffering agents, tonicity adjusting agents, wetting agents, or detergents, among others, to approximate physiological conditions.In some embodiments, the composition is formulated for oral delivery; for example, the composition may contain inactive ingredients such as binders or fillers for the formulation of unit dosage forms such as tablets or capsules.In some embodiments, the composition is formulated for topical administration; for example, the composition may contain inactive ingredients such as solvents or emulsifiers for the formulation of creams, ointments, gels, pastes, or eye drops.

[0232] The compositions may be sterilized, for example, by conventional sterilization techniques or sterile filtered. Aqueous solutions may be packaged ready to use or lyophilized, with the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the preparation will generally be 3-11, more preferably 5-9 or 6-8, most preferably 6-7, e.g., 6-6.5. In some embodiments, the solid form of the composition is packaged in a plurality of single-dose units, each containing a fixed amount of one or more of the above-mentioned agents, such as a sealed package of tablets or capsules. In some embodiments, the solid form of the composition is packaged in a flexible quantity container, such as a squeezable tube designed for a topically applicable cream or ointment.

[0233] In some embodiments, the amount of [ 225 Compositions comprising the α-Ac]-radioimmunoconjugate are provided.

[0234] kit In some embodiments, (1) the method of claim 1, 225 and (2) instructions for administering the composition in combination with a checkpoint inhibitor.

[0235] In some embodiments, the present invention relates to a composition comprising (1) a checkpoint inhibitor and (2) a method for treating a pulmonary artery disease as described herein. 225 and instructions for administering the composition in combination with the [Ac]-radioimmunoconjugate are provided.

[0236] effect In some embodiments, the method of the present disclosure provides a therapeutic effect. In some embodiments, the therapeutic effect comprises an immune response, for example, the immune response comprises an increase in T cells, for example, CD8+ (e.g., IFNγ-producing CD8+ cells) and / or CD4+ cells. In some embodiments, the T cells comprise T cells specific for tumor-associated or tumor-specific antigens expressed in the cancer being treated or improved. In some embodiments, an increase in T cells is observed in the tumor compared to the spleen.

[0237] In some embodiments, the administering step results in at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, or at least 70% of the total T cell population in the mammalian sample being specific for a tumor-associated or tumor-specific antigen, hi some embodiments, the sample is a tumor sample.

[0238] In some embodiments, the therapeutic effect comprises a reduction in tumor volume (e.g., at least partial tumor regression), a stable tumor volume, or a decrease in the rate of growth of tumor volume. In some embodiments, the therapeutic effect comprises a decrease in the occurrence of recurrence or metastasis.

[0239] In some embodiments, the therapeutic effect comprises tumor regression, i.e., a reduction in tumor volume. In some embodiments, tumor regression is characterized by a reduction in tumor volume of at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the tumor volume before the start of treatment. In some embodiments, the therapeutic effect comprises complete tumor regression.

[0240] In some embodiments, tumor regression (whether partial or complete) is persistent in that tumor volume does not substantially increase again after decreasing for a period of time.In some embodiments, tumor regression is persistent for at least 5 days, at least 10 days, at least 15 days, at least 20 days, at least 23 days, at least 25 days, at least 26 days, at least 27 days, at least 28 days, at least 29 days, or at least 30 days after treatment initiation. Other drugs

[0241] In some embodiments, the disclosed methods further comprise administration of an antiproliferative agent, a radiosensitizer, or an immunosuppressant or immunomodulatory agent.

[0242] "Antiproliferative" or "antiproliferative agent" are used interchangeably herein and refer to anti-cancer agents, including those listed in Table 2, any of which may be used in combination with radioimmunoconjugates to treat a condition or disorder. Antiproliferative agents also include organoplatinum derivatives, naphthoquinone and benzoquinone derivatives, chrysophanic acids and their anthraquinone derivatives.

[0243] "Immunoregulatory agent" or "immunomodulatory agent" are used interchangeably herein and refer to immunomodulatory factors, including those listed in Table 2, any of which may be used in combination with the radioimmunoconjugates.

[0244] As used herein, "radiosensitizer" includes any agent that enhances the sensitivity of cancer cells to radiation therapy.Radiosensitizers can include, but are not limited to, 5-fluorouracil, platinum analogs (e.g., cisplatin, carboplatin, oxaliplatin), gemcitabine, EGFR antagonists (e.g., cetuximab, gefitinib), farnesyltransferase inhibitors, COX-2 inhibitors, bFGF antagonists, and VEGF antagonists. Table 2 [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] EXAMPLES

[0245] Example 1. Single-agent efficacy of checkpoint inhibitors in the CT-26 syngeneic model was observed. Single-agent efficacy testing of two checkpoint inhibitors (PD-1 and CTLA-4) was performed in the CT-26 model, a mouse colon cancer model. These carcinomas are known to be partially sensitive to α-PD-1mAb and sensitive to α-CTLA-4mAb. Mice were injected intraperitoneally with either 5 or 15mg / kg of either α-PD-1mAb or α-CTLA-4mAb. The α-PD-1mAb group was administered twice weekly for 4 weeks. The α-CTLA-4mAb group was administered only three times a day with an interval of 3 days. As expected in this model, CTLA-4 treatment was more effective than PD-1 treatment. In both treatment groups, 5mg / kg appeared to be the dose that most effectively impaired tumor growth. See Figure 1. Recruitment of CD8+ / CD4+ T cells after different treatments is also measured using immunohistochemistry and flow cytometry techniques.

[0246] Example 2. Selection of hIGF-1R-expressing CT26 clones to generate mouse cancer models CT26 cells were stably transfected with human IGF-1R plasmid. For selection of hIGF-1R expressing clones, Western blot analysis was performed for the presence of hIGF-1R. See Figure 7. The best clones were selected based on both in vitro and in vivo characteristics. The resulting cell lines were xenografted into mice to express the human IGF-1R, as further described below. 225 Ac] Compound D (conjugated with Compound A1, [ 225A mouse model was generated to test further synergy with TAB-199 radiolabeled with [Ac]; see Example 5-B) and / or other radioimmunoconjugates (e.g., conjugates containing AVE1642) and immune checkpoint inhibitors.

[0247] Example 3. CT-26 Syngeneic Model 177 Lu]-Biodistribution of Compound B MAB391, a mouse monoclonal antibody against IGF-1R (see, e.g., FJ Calzone et al., PLoS One. 2013;8(2):e55135), was conjugated with Compound A1 (a bifunctional chelate represented by the structure shown below) and radiolabeled with Lu-177 using methods well known in the art to obtain [ 177 Lu]-Compound B was formed. [ka]

[0248] Targeting antigen-expressing mouse IGF-1R-overexpressing tumors in vivo 177 The efficacy of [Lu]-Compound B was demonstrated using a CT-26 syngeneic model. Tumor uptake was stable at 15-17% injected dose / g (ID / g) from 24-96 hours post-injection. See Figure 2.

[0249] Example 4. In immunocompetent mice compared to immunodeficient mice 225 Ac]-Enhanced efficacy of Compound C MAB391, a mouse monoclonal antibody against IGF-1R, was conjugated to compound A1 using standard techniques. 225 Ac] and radiolabeled with [ 225 Ac]-Compound C. 225 The efficacy of compound C was tested using 92.5 kBq / kg or 740 kBq / kg (50 nCi or 400 nCi) doses of [ 225 Ac]-Compound C was used.225 Ac]-Compound C was found to have enhanced efficacy in reducing tumor volume in mice with intact immune systems compared to mice without immune systems. See Figure 3.

[0250] Notably, a dose of 50 nCi in mice corresponds to 92.5 kBq / kg, which is equivalent to about 7 MBq in humans (human equivalent dose), and a dose of 400 nCi in mice corresponds to 740 kBq / kg, which is equivalent to about 55 MBq in humans (human equivalent dose).

[0251] Example 5 - A. CT26 Syngeneic Mouse Model 225 Ac]-Synergy between Compound C and α-CTLA-4 / PD-1 treatment. In vivo synergy studies were performed to assess the effect of [ 225 The effects of [Ac]-Compound C (described in Example 4) and checkpoint inhibitors, α-CTLA-4 and α-PD-1 antibodies, were tested. Mice treated with either a CTLA-4 inhibitor alone or a PD-1 inhibitor alone showed a modest reduction in relative tumor volume when compared to the vehicle control group. A dose of 370 kBq / kg (or 200 nCi) of [ 225 Mice treated with [Ac]-Compound C showed a greater reduction in tumor volume compared to the vehicle control group or groups receiving either the CTLA-4 inhibitor or the PD-1 inhibitor alone. 225 Ac]- Compound C was administered at a dose of 370 kBq / kg with either CTLA-4 or PD-1, or both, and showed synergistic effects. 225 Ac]-Compound C, or compared to treatment with a CTLA-4 inhibitor or a PD-1 inhibitor alone, resulted in significantly smaller tumor volumes. See Figure 4A.

[0252] Of note, a dose of 200 nCi in mice corresponds to 370 kBq / kg, which is equivalent to approximately 28 MBq in humans (human equivalent dose).

[0253] Example 5-B. CT26 Syngeneic Mouse Model 225 Ac]-Synergy between Compound D and α-CTLA-4 / PD-1 treatment. TAB-199, a human monoclonal antibody against IGF-1R (see, e.g., https: / / www.antibodypedia.com / gene / 4140 / IGF1R / antibody / 2726933 / TAB-199), was conjugated to Compound A1 using standard techniques known in the art, 225 Ac] and radiolabeled with [ 225 Ac]-Compound D was formed.

[0254] In vivo synergy studies were performed to assess the effect of [ 225 The effects of [Ac]-Compound D and checkpoint inhibitors, α-CTLA-4 and α-PD-1 antibodies, were examined. 225 Mice treated with [Ac]-Compound D showed only transient tumor regression followed by tumor regrowth. 225 When Compound D was co-administered at 370 kBq / kg with either CTLA-4 or PD-1, or both, synergistic effects were observed, demonstrating sustained tumor regression, and co-administration was associated with a significant improvement in [ 225 Ac]-Compound D resulted in significantly smaller tumor volumes. See Figure 4B.

[0255] Of note, a dose of 200 nCi in mice corresponds to 370 kBq / kg, which is equivalent to approximately 28 MBq in humans (human equivalent dose).

[0256] Two days before and 12 days after treatment initiation, 200 nCi[ 225Blood samples were taken from mice treated with [Ac]-compound D and analyzed for T cell receptor repertoire using ImmunoSEQ technology (see, e.g., Wolf K, DiPaolo D., Immunosequencing: accelerating discovery in immunology and medicine. Curr Trends Immunol 2016;17:85-93; Liu X, Wu J. History, applications, and challenges of immune repertoire research. Cell Biol Toxicol 2018;34(6):441-57). The results showed that [ 225 These results suggest that treatment with Ac]-Compound D induces a more clonal T cell response suggestive of immune activation induced by immune complexes.

[0257] Example 6. CT26 rechallenge 225 Development of protective immunity in mice re-treated with Ac]-compound C A rechallenge experiment was performed to determine the effect of the CT26 rechallenge 225 The development of protective immunity in mice treated with [Ac]-Compound C was examined. 225 Ac]-previously treated with Compound C alone or in combination with α-CTLA-4 or α-PD-1 antibodies. Naive mice were used as controls. 225 All mice previously treated with [Ac]-Compound C+ / -anti-CTLA-4 or anti-PD-1 antibodies were protected from tumor challenge, suggesting the development of protective T cell immunity. See Figure 5.

[0258] Example 7. 225 Cytokine responses and T cell recruitment following Ac]-Compound C treatment [ 225 Cytokine responses and T cell recruitment were measured after treatment with [Ac]-Compound C. Mice were administered 1 × 10 6 CT26 cells were inoculated into the mice. 225Mice were treated with either [Ac]-Compound C, unconjugated MAB391 antibody or vehicle. Samples from tumor, spleen and plasma were analyzed for the presence of cytokines at 24, 48 or 72 hours. Additional samples were taken from tumor and spleen at 72 hours, 5 days and 8 days for immunohistochemistry to evaluate the presence of different T cell types. Finally, on day 8, tumor-infiltrating lymphocytes were extracted, isolated and quantified using flow cytometry. See Figure 6.

[0259] As shown in Table 3 below, compared to the unconjugated MAB391 antibody, 225 Changes in cytokine expression were observed in tumors treated with Ac]-Compound C: Table 3. Changes in cytokine expression [Table 3]

[0260] Example 8. Combination therapy leads to an increase in tumor-associated antigen-specific CD8+ T cells in both the spleen and the tumor itself. [ 225 Ac]-Compound D (see Example 5-B) is a compound that reacts with actinium-225 ( 225 This is a radioimmunoconjugate containing the human monoclonal IGF-1R antibody TAB-199 labeled with Ac). 225 Combinations of [Ac]-Compound D with checkpoint inhibitors (α-PD-1, α-CTLA-4, or both α-PD-1 and α-CTLA-4) were tested in the CT26 syngeneic mouse model. Mice were rechallenged with CT26 cells 28 days after initial tumor inoculation.

[0261] CD8+ and CD4+ T cell populations were assessed in both the spleen and tumor after rechallenge. 225In mice treated with Ac]-Compound D and checkpoint inhibitors, both spleen and tumor showed the presence of CD8+ T cells. Importantly, an increase in the frequency of CD8+ T cells was observed in tumors compared to controls. These results suggest that these combined treatments result in improved levels of therapeutically effective CD8+ T cells.

[0262] Antigen-specific T cells were detected and enumerated using an MHC class I tetramer assay. In this assay, MHC I molecules presenting epitopes specific for CT26 cells are labeled with biotin. In the presence of streptavidin, these MHC I molecules tetramerize. CD8+ T cells specific for the CD26 epitope are thereby labeled when their T cell receptors bind to the MHC I / CT26 epitope complex in the tetramer. Based on the tetramer analysis, approximately 35%, 62%, and 75% of CD8+ T cells, respectively, were [ 225 Ac]-Compound D / α-CTLA-4, [ 225 Ac]-Compound D / α-PD-1 and [ 225 Ac]-Compound D / α-CTLA-4 / α-PD-1 treated mice were antigen-specific.

[0263] Example 9. CT26 syngeneic mouse model 225 Ac]-Synergy between Compound D1 and α-CTLA-4 / PD-1 treatment. TAB-199 was conjugated to Compound A2 (a bifunctional chelate represented by the structure shown below) using standard techniques known in the art: 225 Ac] and radiolabeled with [ 225 Ac]-to form compound D1. [ka]

[0264] In vivo synergy studies were performed to assess the effect of [ 225The effects of [Ac]-Compound D1 and checkpoint inhibitors, α-CTLA-4 and α-PD-1 antibodies, were examined. 225 Mice treated with [225Ac]-Compound D1 showed only transient tumor regression followed by tumor regrowth. However, when [225Ac]-Compound D1 was co-administered at 370 kBq / kg with either CTLA-4 or PD-1, or both, a synergistic effect was observed, showing sustained tumor regression, and co-administration was associated with a significant increase in [ 225 Treatment with 100-mL guinea pigs containing 100% guinea pigs resulted in significantly smaller tumor volumes when compared to treatment with 100-mL [Ac]-Compound D1. See Figure 9A.

[0265] Of note, a dose of 200 nCi in mice corresponds to 370 kBq / kg, which is equivalent to approximately 28 MBq in humans (human equivalent dose).

[0266] Example 10. CT26 syngeneic mouse model 225 Ac]-Synergy between Compound D2 and α-CTLA-4 / PD-1 treatment. TAB-199 was conjugated to Compound A3 (a bifunctional chelate represented by the structure shown below) using standard techniques known in the art: 225 Ac] and radiolabeled with [ 225 Ac]-to form compound D2. [ka]

[0267] In vivo synergy studies were performed to assess the effect of [ 225 The effects of [Ac]-Compound D2 and checkpoint inhibitors, α-CTLA-4 and α-PD-1 antibodies, were examined. 225Mice treated with [225Ac]-Compound D2 showed only transient tumor regression followed by tumor regrowth. However, when [225Ac]-Compound D was co-administered at 370 kBq / kg with either CTLA-4 or a combination of PD-1 and CTLA-4, a synergistic effect was observed, showing sustained tumor regression, and co-administration was associated with a significant increase in [ 225 Ac]-Compound D2 treatment resulted in significantly smaller tumor volumes. See Figure 9B.

[0268] Of note, a dose of 200 nCi in mice corresponds to 370 kBq / kg, which is equivalent to approximately 28 MBq in humans (human equivalent dose).

[0269] Example 11. AVE1642 included 225 Effect of combination therapy with Ac]-labeled conjugates. AVE1642 is conjugated to a bifunctional chelate, such as Compound A1, Compound A2, or Compound A3, and then 225 Ac], including AVE1642 (a humanized monoclonal IGF-1R antibody), which can be prepared by radioactive labeling with [ 225 [Ac]-labeled radioimmunoconjugates can be tested in combination with checkpoint inhibitors (e.g., CTLA-4 and / or PD-1 antibodies) using protocols similar to those described in Examples 4-9. For example, effects on tumor volume, animal survival, cytokine expression, T cell immunity (e.g., the presence, quantity and / or function of tumor-associated antigen-specific CD8+ T cells), and protection against tumor rechallenge can be compared between combination therapy and monotherapy treatment and / or control groups.

[0270] Example 12. A peptide comprising an FGFR3 targeting moiety 225 Effect of combination therapy with Ac]-labeled conjugates. The FGFR3 targeting moiety is conjugated to a bifunctional chelate, such as Compound A1, Compound A2, or Compound A3, and then 225

[0036] The FGFR3 targeting moiety (e.g., an FGFR3 antibody or fragment thereof, or a small molecule) can be prepared by radioactive labeling with [ 225 [Ac]-labeled conjugates can be tested in combination with checkpoint inhibitors (e.g., CTLA-4 and / or PD-1 antibodies) using mouse models of FGFR3-altered cancers using experiments similar to those described in Examples 4-9. For example, effects on tumor volume, animal survival, cytokine expression, T cell immunity (e.g., the presence, quantity and / or function of tumor-associated antigen-specific CD8+ T cells), and protection against tumor rechallenge can be compared between combination therapy and monotherapy treatment and / or control groups. Equivalents / Alternative Embodiments

[0271] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments described herein which equivalents are intended to be encompassed by the following claims.

Claims

1. 1. A method of treating a patient having cancer, comprising: (i) administering to the patient [ ２２５ Administering a radioimmunoconjugate comprising a medicament for treating a patient with rheumatoid arthritis, the ... (ii) administering to the patient one or more checkpoint inhibitors, wherein the patient ２２５ have received or are receiving a [Ac]-radioimmunoconjugate, or (iii) in combination with one or more checkpoint inhibitors, ２２５ administering to said patient a radioimmunoconjugate comprising: The above [ ２２５ Ac]-radioimmunoconjugate having the formula: A-L １ -XL ２ -Z-B ２２５ Ac, wherein A is DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), DOTMA (1R,4R,7R,10R)-α,α',α",α"'-tetramethyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid, DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane), DO3AM-acetic acid (2-(4,7,10-tris(2-amino-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid), a chelating moiety selected from the group consisting of 10-(2-hydroxypropyl)-1,4,7-tetraazacyclododecane-1,4,7-triacetic acid), DOTP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonic acid)), DOTA-4AMP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakis(acetamido-methylenephosphonic acid), NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid), and HP-DO3A (10-(2-hydroxypropyl)-1,4,7-tetraazacyclododecane-1,4,7-triacetic acid); L １ is a bond or an optionally substituted C １～６ Alkyl or C １～６ is heteroalkyl, X is -C(O)NR １ -*, -NR １ C(O)-*, -OC(O)NR １ -*, -NR １ C(O)O-*, -NR １ C(O)NR １ --, --CH ２ -Ph-C(O)NR １ -*, -NR １ C(O)-Ph-CH ２ -*, -O-, or -NR １ -, where "*" is L ２ Each R １ are independently hydrogen or C １～６ is alkyl, L ２ is optionally substituted C １～５０ Alkyl or C １～５０ is heteroalkyl, Z is -C(O)-, -CH ２ -, -OC(O)-#, -C(O)O-#, -NR ２ C(O)-#, -C(O)NR ２ -# or -NR ２ -, where "#" indicates the point of attachment to B, and each R ２ are independently hydrogen or C １～６ is alkyl, B is a targeting moiety, The above [ ２２５ Administering to said patient a dose of 10 kBq to 400 kBq per kg of body weight of said patient or as a unit dose of 1 to 30 MBq of said radioimmunoconjugate.

2. The patient is ２２５ 2. The method of claim 1, comprising administering a [C6-Ac]-radioimmunoconjugate to a patient having a pulmonary circulation that is refractory to pulmonary circulation, wherein the patient has received or is receiving one or more checkpoint inhibitors.

3. In combination with one or more checkpoint inhibitors ２２５ 2. The method of claim 1, comprising administering to the patient a [Cas9]HLA-CoA [Cas9] radioimmunoconjugate.

4. The method of any one of claims 1 to 3, wherein the chelating moiety is DOTA.

5. The compound has formula I: 【Chemical 34】 The method according to any one of claims 1 to 3, wherein

6. The compound has formula II: 【Chemistry 35】 The method according to any one of claims 1 to 3, wherein

7. The method of any one of claims 1 to 3, wherein the targeting moiety comprises an antibody or an antigen-binding fragment thereof.

8. 8. The method of claim 7, wherein B is an insulin-like growth factor 1 receptor (IGF-1R) antibody or an antigen-binding fragment thereof, an endosialin (TEM-1) antibody or an antigen-binding fragment thereof, or a fibroblast growth factor receptor 3 (FGFR3) antibody or an antigen-binding fragment thereof.

9. 9. The method of claim 8, wherein B is an IGF-1R antibody or an antigen-binding fragment thereof selected from the group consisting of figitumumab, cixutumumab, TAB-199, AVE1642, BIIB002, lobatumumab, and teprotumumab, and antigen-binding fragments thereof.

10. 10. The method of claim 9, wherein B is AVE1642 or an antigen-binding fragment thereof.

11. The above [ ２２５ The method of any one of claims 1 to 3, wherein the [Ac]-radioimmunoconjugate is administered at a dose of about 10 kBq to about 200 kBq per kg of body weight of the patient.

12. The above [ ２２５ The method of any one of claims 1 to 3, wherein the [Ac]-radioimmunoconjugate is administered at a dose of about 30 kBq to about 120 kBq per kg of body weight of the patient.

13. 4. The method of any one of claims 1-3, wherein the one or more checkpoint inhibitors comprise a PD-1 inhibitor, a CTLA-4 inhibitor, or a combination thereof.

14. 14. The method of claim 13, wherein the one or more checkpoint inhibitors comprise both a PD-1 inhibitor and a CTLA-4 inhibitor.

15. The method of claim 13, wherein the PD-1 inhibitor or the CTLA-4 inhibitor is an antibody.

16. The method of any one of claims 1 to 3, wherein the one or more checkpoint inhibitors are administered at a lower effective dose.

17. The above [ ２２５ The method of any one of claims 1 to 3, wherein the [Ac] radioimmunoconjugate is administered at a lower effective dose.

18. 4. The method of any one of claims 1-3, wherein the one or more checkpoint inhibitors comprises a PD-1 inhibitor administered at a dose of about 5 mg / kg to about 15 mg / kg.

19. 14. The method of claim 13, wherein the PD-1 inhibitor is pembrolizumab.

20. 4. The method of any one of claims 1-3, wherein the one or more checkpoint inhibitors comprise both a PD-1 inhibitor and a CTLA-4 inhibitor, each administered at a dose of about 5 mg / kg to about 15 mg / kg.

21. 10. The method of claim 9, wherein B is AVE1642 or an antigen-binding fragment thereof, and the one or more checkpoint inhibitors comprise a PD-1 inhibitor that is pembrolizumab.

22. The above [ ２２５ 22. The method of claim 21, wherein the [Ac] radioimmunoconjugate is administered at a dose of about 30 kBq / kg to about 120 kBq / kg of the patient's body weight and the PD-1 inhibitor is administered at a dose of about 5 mg / kg to about 15 mg / kg.

23. 4. The method of any one of claims 1 to 3, wherein the patient has a cancer selected from the group consisting of breast cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, colorectal cancer, cervical cancer, endometrial cancer, sarcoma, adrenocortical carcinoma, neuroendocrine cancer, Ewing's sarcoma, multiple myeloma, and acute myeloid leukemia.

24. The method of any one of claims 1 to 3, wherein the patient has a solid tumor that expresses IGF-1R.

25. The method according to any one of claims 1 to 3, wherein B is capable of binding to a tumor-associated antigen and said administration results in an expansion of CD8+ T cells specific for said tumor-associated antigen.

26. 26. The method of claim 25, wherein the administering step results in at least 60% of the total CD8+ T cell population in the patient-derived sample being specific for the tumor-associated antigen.

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

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