Combination therapy using radioimmunoconjugate and inhibitor of DNA damage and repair

Abstract

To provide combination therapy comprising administering a radioimmunoconjugate and an inhibitor of DNA damage response.SOLUTION: The present disclosure encompasses the insight that combining inhibition of DNA damage-repair mechanisms with a therapy that targets DNA breaks to cancer cells may provide a less toxic therapy with improved efficacy. Radioactive decay can cause direct physical damage (such as single or double-stranded DNA breaks) or indirect damage (such as by-stander or crossfire effects) to the biomolecules that constitute a cell. The disclosure combines a radioimmunoconjugate targeting cancer cells with an inhibition of DNA damage-repair to treat or ameliorate cancer.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 774,847, filed December 3, 2018, the entire contents of which are incorporated herein by reference for all purposes. [Background technology]

[0002] DNA single- and double-strand breaks occur for a variety of reasons, including mutations in pathways involving BRCA, PTEN, and ATR proteins. Such DNA breaks are repaired by multiple pathways. PARP inhibition (PARPi) leads to the accumulation of single- and / or double-strand breaks. Existing PARP inhibitors act both by inhibiting PARP enzyme activity and by trapping PARP proteins within chromatin ("DNA trapping"). Tumor cells with BRCA and / or PTEN mutations are sensitive to PARPi, whereas ATR inhibition (ATRi) leads to a failure of double-strand break repair, thus resulting in the accumulation of double-strand breaks. Increased single- or double-strand DNA breaks increase cell death.

[0003] DNA damage repair inhibitors (DDRi) have been investigated as cancer therapeutics. However, inhibiting DNA strand break repair has been observed to increase normal tissue toxicity, and many DDRi have only moderate efficacy in vivo. Improved cancer treatments are needed, particularly those that increase efficacy without increasing toxicity in patients. Summary of the Invention [Means for solving the problem]

[0004] The present disclosure encompasses the insight that combining inhibition of DNA damage repair mechanisms with treatments that target DNA breaks in cancer cells may provide improved efficacy and less toxic therapies. Radioactive decay can cause direct physical damage (e.g., single- or double-stranded DNA breaks) or indirect damage (e.g., bystander or crossfire effects) to the biomolecules that make up cells. The present disclosure combines radioimmunoconjugates that target cancer cells with DNA damage repair inhibition to treat or ameliorate cancer.

[0005] In one aspect, a method for treating or ameliorating cancer is provided, the method comprising the steps of: (i) administering a radioimmunoconjugate to a mammal, wherein the mammal has been administered or is being administered a DNA damage response inhibitor (DDRi); (ii) administering the DDRi to the mammal, wherein the mammal has been administered or is being administered the radioimmunoconjugate; or (iii) administering the DDRi to the mammal simultaneously with administering the radioimmunoconjugate to the mammal.

[0006] In some embodiments, the method comprises administering a DDRi to a mammal, wherein the mammal has been or is being administered a radioimmunoconjugate.

[0007] In some embodiments, the DDRi is administered at a lower effective dose. In some embodiments, the radioimmunoconjugate is administered at a lower effective dose. In some embodiments, both the DDRi and the radioimmunoconjugate are administered at lower effective doses.

[0008] In some embodiments, the radioimmunoconjugate comprises (i) a targeting moiety, (ii) a linker, and (iii) a chelating moiety or a metal complex of a chelating moiety.

[0009] In some embodiments, the targeting moiety is capable of binding to a tumor-associated antigen. In some embodiments, the tumor-associated antigen is a tumor-specific antigen.

[0010] In some embodiments, the targeting moiety is an antibody or an antigen-binding fragment thereof.

[0011] In some embodiments, the antibody or antigen-binding fragment thereof is an IGF1-R antibody or antigen-binding fragment thereof.

[0012] In some embodiments, the antibody or antigen-binding fragment thereof is an endosialin (TEM-1) antibody or antigen-binding fragment thereof.

[0013] In some embodiments, the radioimmunoconjugate comprises a metal complex of a chelating moiety. In some embodiments, the metal complex comprises a radionuclide. In some embodiments, the radionuclide is an alpha emitter, e.g., astatine-211 ( 211 At), Bismuth-212( 212 Bi), Bismuth-213( 213 Bi), Actinium-225( 225 Ac), radium-223( 223 Ra), lead-212( 212 Pb), thorium-227( 227 Th), and terbium-149 ( 149 In some embodiments, the radionuclide is an alpha emitter selected from the group consisting of: 225 It is Ac.

[0014] In some embodiments, the radioimmunoconjugate has the following structure: [ka] where B is a targeting moiety. Includes.

[0015] In some embodiments, the DDRi is a PARP inhibitor.In some embodiments, the PARP inhibitor is a small molecule PARP inhibitor, for example, a PARP inhibitor selected from the group consisting of niparib, niraparib, olaparib, talazoparib, pamiparib, rucaparib (camsylate) and veliparib, or analogs thereof.In some embodiments, the small molecule PARP inhibitor is olaparib or analogs thereof.

[0016] In some embodiments, the DDRi is an ATR inhibitor. In some embodiments, the ATR inhibitor is a small molecule ATR inhibitor, for example, the ATR inhibitor is selected from the group consisting of AZ20, AZD0156, AZD1390, AZD6738, BAY-1895344, EPT-46464, M3541, M4344, M6620 (formerly known as VE-922 or VX-970), NU6027, VE-821, or analogs thereof. In some embodiments, the ATR inhibitor is BAY-1895344 or an analog thereof.

[0017] In some embodiments, the DDRi is a WEE1 inhibitor, a Chk1 inhibitor, or a Chk2 inhibitor.

[0018] In some embodiments, the DDRi is a DNA protein kinase (DNA-PK) inhibitor.

[0019] In some embodiments, the mammal is a human.

[0020] In some embodiments, the mammal has been diagnosed with cancer.

[0021] In some embodiments, the cancer is 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, sarcoma, adrenocortical carcinoma, neuroendocrine carcinoma, Ewing's sarcoma, multiple myeloma, or acute myeloid leukemia.

[0022] In some embodiments, the mammal has at least one solid tumor.

[0023] In some embodiments, the administration results in a therapeutic effect. In some embodiments, the therapeutic effect comprises a decrease in tumor volume, a stable tumor volume, or a decrease in the rate of tumor volume growth. In some embodiments, the therapeutic effect comprises a decrease in the incidence of recurrence or metastasis.

[0024] definition Chemical terminology: The term "acyl," as used herein, refers to a hydrogen or alkyl group (e.g., a 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., a 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.

[0025] The term "alkyl," as used herein, unless otherwise specified, includes both straight-chain 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 of 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) -CO2R optionally substituted with an O-protecting group A’ , where R A’ is (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 (wherein 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 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 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 of which is independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C6-10 (16) -SO2R D’ , where R D’ is (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 alk-C 6-10 (17) -SO2NR E’ R F’ , where R E’ and R F’ each of which is independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 Aryl and (d) C 1-6 alk-C 6-10 (18) -C(O)R G’ , where R G’ is (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 (wherein 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 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 each R N1are 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 2-20 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 (wherein 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 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 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 1-6alkyl), (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) s3 OR' polyethylene glycol (wherein 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 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 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 a C-alkaryl can be further substituted with an oxo group to provide the respective aryloyl substituent.

[0026] 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.

[0027] The term "alkenyl," unless otherwise specified, 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, and is 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 can 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.

[0028] 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, and is 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.

[0029] As used herein, the term “amino” refers to —N(R N1 )2, wherein each RN1 are independently H, OH, NO2, N(R N2 )2, SO2OR N2 , SO2R N2 , SOR N2 , an N-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, alkaryl, cycloalkyl, alkcycloalkyl, carboxyalkyl (e.g., optionally substituted with an O-protecting group, e.g., optionally substituted with an arylalkoxycarbonyl group or any described herein), sulfoalkyl, acyl (e.g., acetyl, trifluoroacetyl, or others described herein), alkoxycarbonylalkyl (e.g., optionally substituted with an O-protecting group, e.g., optionally substituted with an arylalkoxycarbonyl group or any described herein), heterocyclyl (e.g., heteroaryl), or 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, and each R N2 are independently H, alkyl, or aryl. An amino group can 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 —NH 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 is H, C 1-20 Alkyl (e.g., C 1-6 alkyl), or C6-10 It may be aryl.

[0030] As used herein, "amino acid" refers to a molecule having a side chain, an amino group, and an acid group (e.g., a carboxy group of -COH or a sulfo group of -SOH), where the amino acid is attached to the parent molecular group by the side chain, amino group, or 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 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 optionally be substituted with one, two, three, or, in the case of amino acid 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 (heterocyclyl)oxy; (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’ is (a)C 1-20Alkyl (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 (wherein 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 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 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 of which is independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C 1-6 alk-C 6-10 (16) -SO2R D’ , where R D’ is (a)C 1-6 Alkyl, (b) C 6-10 Aryl, (c) C 1-6 alk-C 6-10 (17) -SO2NR E’R F’ , where R E’ and R F’ each of which is independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 Aryl and (d) C 1-6 alk-C 6-10 (18) -C(O)R G’ , where R G’ is (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 (wherein 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 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 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 1-20Alkyl (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) s3 OR' polyethylene glycol (wherein 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 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 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) s3 OR' polyethylene glycol (wherein 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 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 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.

[0031] 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 1-6Alkyl (e.g., perfluoroalkyl), hydroxy-C 1-6 Alkyl, nitro-C 1-6 Alkyl, or C 1-6 Thioalkoxy-C 1-6 alkyl); (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, such as 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 between 0 and 4, and R A’ is (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 between 0 and 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 6-10aryl, and (c) 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 of which is independently (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 a C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to give the respective aryloyl and (heterocyclyl)oyl substituents.

[0032] 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, and "alk" refers to any group selected from the group consisting of C, C- ... 1-6 refers to alkylene, the chemical structure to which it is attached is as defined herein.

[0033] The term "carbonyl" as used herein refers to a C(O) group, which may also be represented as C=O.

[0034] The term "carboxy" as used herein means -CO2H.

[0035] As used herein, the term "cyano" refers to a -CN group.

[0036] As used herein, unless otherwise specified, "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, bicycloheptyl, 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 alkyl); (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, such as 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 between 0 and 4, and R A’ is (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 between 0 and 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’ is (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 of which is independently (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 a C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to give the respective aryloyl and (heterocyclyl)oyl substituents.

[0037] As used herein, the term "diastereomers" means stereoisomers that are not mirror images of one another and are not superimposable with respect to one another.

[0038] 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%.

[0039] The term "halogen" as used herein refers to a halogen selected from bromine, chlorine, iodine, or fluorine.

[0040] As used herein, the term "heteroalkyl" 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. As used herein, the terms "heteroalkenyl" and "heteroalkynyl" refer to alkenyl and alkynyl groups, 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 and heteroalkynyl groups can be further substituted with one, two, three, or four substituents as described herein for alkyl groups.

[0041] The term "heteroaryl," as used herein, refers to a subset of heterocyclyl, as defined herein, that is aromatic, i.e., that contains 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.

[0042] "Heteroarylalkyl," as defined herein, 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.

[0043] As used herein, unless otherwise specified, the term "heterocyclyl" 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 to 2 double bonds, and 6- and 7-membered rings have 0 to 3 double bonds. Exemplary unsubstituted heterocyclyl 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. 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 carbon 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 are 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-amino5-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 Zolyl (e.g., 1-(ethoxycarbonyl)-1H-benzopyrazolyl); 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. [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 optionally be 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 alkyl); (3) C 1-20 Alkoxy (e.g., C 1-6 Alkoxy, such as perfluoroalkoxy; (4) C 1-6 Alkylsulfinyl; (5)C 6-10 Aryl;(6)Amino;(7)C 1-6alk-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 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 between 0 and 4, and R A’ is (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)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 of which is independently (a) hydrogen, (b) C 1-6 Alkyl, (c) C 6-10 aryl, and (d) C1-6 alk-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 a C1-alkaryl or C1-alkheterocyclyl can be further substituted with an oxo group to give the respective aryloyl and (heterocyclyl)oyl substituents.

[0044] As used herein, the term "hydrocarbon" refers to groups consisting solely of carbon and hydrogen atoms.

[0045] As used herein, the term "hydroxyl" 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.

[0046] As used herein, the term "isomer" 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 can therefore 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 diastereoisomerically 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.

[0047] The term "N-protected amino" as used herein refers to an amino group, as defined herein, having one or two N-protecting groups, as defined herein, attached thereto.

[0048] As used herein, the term "N-protecting group" 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 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).

[0049] As used herein, the term "O-protecting group" 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-isopropylsilyloxymethyl, 4,4′-dimethoxytrityl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidino, and 4-nitrobenzoyl; alkylcarbonyl groups, such as acyl, acetyl, propionyl, pivaloyl, and the like; and, optionally, substituted arylcarbonyl groups, such as benzoyl; silyl groups, such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-isopropylsilyloxymethyl (TOM), triisopropylsilyl (TIPS); hydroxyl-containing ether-forming groups such as methyl, methoxymethyl, tetrahydropyranyl, benzyl, p-methoxybenzyl, trityl, etc.; alkoxycarbonyl groups, such as methoxycarbonyl, ethoxycarbonyl, isopropoxycarbonyl, n-isopropoxycarbonyl, n-butyloxycarbonyl, isobutyloxycarbonyl, sec-butyloxycarbonyl, t-butyloxycarbonyl, 2-ethylhexyloxycarbonyl, cyclohexyloxycarbonyl, methyloxycarbonyl, etc.;Alkoxyalkoxycarbonyl groups, such as methoxymethoxycarbonyl, ethoxymethoxycarbonyl, 2-methoxyethoxycarbonyl, 2-ethoxyethoxycarbonyl, 2-butoxyethoxycarbonyl, 2-methoxyethoxymethoxycarbonyl, allyloxycarbonyl, propargyloxycarbonyl, 2-butenoxycarbonyl, 3-methyl-2-butenoxycarbonyl, etc.; haloalkoxycarbonyl groups, 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 diphenylmethylsilyl); 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 dimethyl acetal, 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, orthoester, and the like; and oxazoline groups);

[0050] The term "oxo" as used herein refers to =O.

[0051] As used herein, the term "polyethylene glycol" refers to an alkoxy chain consisting of one or more monomer units, each consisting of -OCH2CH2-. Polyethylene glycol (PEG) is sometimes also 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 of 1 to 10 (e.g., 1 to 6 or 1 to 4), and 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). Polyethylene glycol may also have the structure -NR N1 (CH2) s2 (CH2CH2O) s1 (CH2) s3 NR N1-amino-polyethylene glycol, wherein 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 each R N1 are independently hydrogen or optionally substituted C 1-6 It can also be considered to include alkyl.

[0052] As used herein, the term "stereoisomer" refers to all possible different isomeric and conformational forms that a compound (e.g., a compound of any formula described herein) may possess, particularly all stereochemically and conformationally possible isomers, all diastereomers, enantiomers, and / or conformers of the basic molecular structure. Some compounds may exist in different tautomeric forms, and all tautomeric forms are included within the scope of this disclosure.

[0053] The term "sulfonyl" as used herein refers to the group -S(O)2-.

[0054] The term "thiol" as used herein refers to a -SH group.

[0055] Other terms As used herein, the terms "combined administration," "coadministration," or "coadministration" mean that two or more agents are administered to a subject simultaneously or at intervals such that the effects of each agent on the patient can overlap. Thus, two or more agents administered in combination need not 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 day) of each other, within 28 days (e.g., within 14, 7, 6, 5, 4, 3, 2, or 1 day), within 24 hours (e.g., within 12, 6, 5, 4, 3, 2, or 1 hour), or within about 60, 30, 15, 10, 5, or 1 minute. In some embodiments, the administration of the agents is spaced closely enough apart so that a combined effect is achieved.

[0056] As used herein, "administering" an agent to a subject includes contacting cells of said subject with the agent.

[0057] 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. Antibodies can be of any type (e.g., IgA, IgD, IgE, IgG, or IgM) or subtype (e.g., IgA1, IgA2, IgG1, IgG2, IgG3, or IgG4). Those skilled in the art will understand that a characteristic sequence or portion of an antibody can 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, those skilled in the art will understand that a characteristic sequence or portion of an antibody can include one or more polypeptide chains and can include sequence elements found in the same polypeptide chain or different polypeptide chains.

[0058] As used herein, "antigen-binding fragment" refers to a portion of an antibody that retains the binding properties of the parent antibody.

[0059] The terms "bifunctional chelate" or "bifunctional complex," used interchangeably herein, refer to a compound comprising a chelating group or a metal complex thereof, a linker group, and a therapeutic moiety, targeting moiety, or bridging group.

[0060] The term "cancer" refers to any cancer caused by the growth 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. "Hematologic cancers" or "liquid cancers," used interchangeably herein, are cancers that are present in bodily fluids, such as lymphomas and leukemias.

[0061] The term "chelate" refers to an organic compound or moiety thereof that is capable of binding to a central metal or radioactive metal atom at two or more points.

[0062] As used herein, the term "conjugate" refers to a molecule that includes a chelating group or a metal complex thereof, a linker group, and optionally a therapeutic moiety, a targeting moiety, or a bridging group.

[0063] As used herein, the term "compound" is meant to include all stereoisomers, geometric isomers, and tautomers of the structures depicted.

[0064] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise specified, all stereoisomers, such as enantiomers and diastereomers, are intended. Compounds of the present disclosure containing 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 can also be present in the compounds described herein, and all such stable isomers are contemplated in this disclosure. Cis and trans geometric isomers of the compounds of the present disclosure are described and can be isolated as a mixture of isomers or as separate isomeric forms.

[0065] 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 in a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomeric forms can be in equilibrium or sterically locked into one form by appropriate substitution.

[0066] 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 member of such groups or ranges, and every individual subcombination. For example, "C 1-6The term "alkyl" is specifically intended to individually disclose methyl, ethyl, C alkyl, C alkyl, C alkyl, and C alkyl. As used herein, phrases of the form "optionally substituted X" (e.g., optionally substituted alkyl) are intended to be equivalent to "X, wherein 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.

[0067] As used herein, the term "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, an anhydride, a thiol, a disulfide, a maleimide, an azide, an alkyne, a strained alkyne, a strained alkene, a halogen, a sulfonate, a haloacetyl, an amine, a hydrazide, a diazirine, a phosphine, a tetrazine, or an isothiocyanate. In some embodiments, the bridging group can be glycine-glycine-glycine and / or leucine-proline-(any amino acid)-threonine-glycine. These are the recognition sequences for coupling a targeting agent and a linker using sortase-mediated coupling reaction. Those skilled in the art will appreciate that the use of cross-linking groups is not limited to the particular constructs disclosed herein, but rather may include other known cross-linking groups.

[0068] As used herein (e.g., in relation to a treatment outcome or effect), the terms "reduce," "decreased," "increase," "increased," or "decreased" refer to a level 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.

[0069] As used herein, the term "detection agent" refers to a molecule or atom that is useful in diagnosing disease by identifying the location of cells containing antigens.Various methods for labeling polypeptides with detection agents are known in the art.Examples of detection agents include, but are not limited to, radioisotopes and radionuclides, dyes (such as biotin-streptavidin complexes), contrast agents, luminescent agents (such as FITC, rhodamine, lanthanide fluorophores, cyanine, and near-infrared dyes), and magnetic agents such as gadolinium chelates.

[0070] The term "DNA damage and repair inhibitors" (DDRi) refers to drugs that prevent the repair of cellular DNA damage resulting from endogenous or exogenous chromosomal insults, acting through the inhibition of normally occurring DNA repair mechanisms and related processes necessary for maintaining cell viability.

[0071] 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.

[0072] As used herein, the term "immunoconjugate" refers to a complex comprising a targeting moiety, such as an antibody, nanobody, affibody, or fibronectin type III domain consensus sequence. In some embodiments, the immunoconjugate comprises 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).

[0073] 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 present invention when used in conjunction with that agent, and that is lower than the dose determined to be therapeutically effective when that agent is used as a monotherapy in a reference experiment or for other therapeutic guidance.

[0074] The term "pharmaceutical composition" refers to a composition containing a compound described herein formulated with a pharmaceutically 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 treating 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, gel cap, or syrup); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate matter and in a solvent system suitable for intravenous use); or any other formulation described herein.

[0075] As used herein, the term "pharmaceutically acceptable excipient" refers to a component other than the compounds described herein (e.g., a vehicle capable of suspending or dissolving an active compound), which has the properties of being non-toxic and non-inflammatory in patients. Examples of excipients include antiadherents, antioxidants, binders, coating agents, compression aids, disintegrants, dyes (pigments), emollients, emulsifiers, fillers (diluents), film-forming or coating agents, flavorings, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, radiation protectants, adsorbents, suspending or dispersing agents, sweeteners, 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, hydroxypropyl cellulose, 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.

[0076] As used herein, " pharmaceutically acceptable salt " refers to the salt of the compound described herein, which 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.Salt can be prepared in situ during the final isolation and purification of the compound described herein, or by reacting free base group with suitable organic acid separately.

[0077] Compounds may have ionizable groups so that they can be prepared as pharmaceutically acceptable salts.These salts may be acid addition salts containing inorganic or organic acids, or in the case of acidic forms of compounds, 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.

[0078] 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, and 2-hydroxy-ethoxybenzoate, among others.

[0013] Representative salts of 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.

[0079] As used herein, the term "polypeptide" 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. Those skilled 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.

[0080] As used herein, the term "radioconjugate" refers to any complex containing a radioisotope or radionuclide, such as any of the radioisotopes or radionuclides described herein.

[0081] 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.

[0082] As used herein, the term "radioimmunotherapy" refers to a method of using a radioimmunoconjugate to produce a therapeutic effect. In some embodiments, radioimmunotherapy can 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 can include administering a radioimmunoconjugate to a cell, where administration of the radioimmunoconjugate kills the cell. Where radioimmunotherapy involves selective killing of a cell, in some embodiments, the cell is a cancer cell in a subject with cancer.

[0083] As used herein, the term "radionuclide" refers to an atom that can undergo 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.

[0084] By "subject" is meant a human or non-human animal (eg, a mammal).

[0085] "Substantial identity" or "substantially identical" refers to a polypeptide sequence that has the same polypeptide sequence as a reference sequence, or a specified percentage of the same 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 typically 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, 1710 University Avenue, Madison, WI 53705). Such software can match similar sequences by assigning degrees of homology to various substitutions, deletions, and other modifications.

[0086] As used herein, the term "targeting moiety" refers to a molecule or any portion of a molecule that binds to a given target. In some embodiments, the targeting moiety is a protein or polypeptide, such as an antibody or antigen-binding fragment thereof, a nanobody, an affibody, or a consensus sequence of a fibronectin type III domain.

[0087] 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, e.g., an antibody, an antigen-binding fragment thereof. In some embodiments, the therapeutic moiety is a small molecule.

[0088] 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 for obtaining a beneficial or desired result, such as a clinical outcome. Beneficial or desired results can 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; improvement or palliation of a disease, disorder, or condition; and remission (partial or complete), whether detectable or undetectable. In the context of cancer treatment, "ameliorating" can include, for example, a reduced incidence of metastasis, a reduction in tumor volume, a reduction in tumor vascularization, and / or a reduced rate of tumor growth. "Alleviating" 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.

[0089] 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.

[0090] As used herein, the term "tumor-specific antigen" refers to an antigen that is endogenously present only in tumor cells. In an embodiment of the present invention, for example, the following items are provided: (Item 1) 1. A method for treating or ameliorating cancer, said method comprising: (i) administering a radioimmunoconjugate to a mammal, wherein the mammal has been or is being administered a DNA damage response inhibitor (DDRi); (ii) administering a DDRi to a mammal, wherein the mammal has been or is being administered a radioimmunoconjugate; or (iii) administering a DDRi to said mammal simultaneously with administering a radioimmunoconjugate to said mammal. A method comprising: (Item 2) 2. The method of claim 1, wherein the method comprises administering a DDRi to a mammal, the mammal having been administered or is being administered a radioimmunoconjugate. (Item 3) 3. The method of item 1 or 2, wherein the DDRi is administered at a lower effective dose. (Item 4) 4. The method of any one of items 1 to 3, wherein the radioimmunoconjugate is administered at a lower effective dose. (Item 5) 2. The method of claim 1, wherein the radioimmunoconjugate comprises (i) a targeting moiety, (ii) a linker, and (iii) a chelating moiety or a metal complex of a chelating moiety. (Item 6) 6. The method of claim 5, wherein the targeting moiety is capable of binding to a tumor-associated antigen. (Item 7) 7. The method of claim 6, wherein the tumor-associated antigen is a tumor-specific antigen. (Item 8) 8. The method of claim 6 or 7, wherein the targeting moiety is an antibody or an antigen-binding fragment thereof. (Item 9) 9. The method of claim 8, wherein the antibody or antigen-binding fragment thereof is an IGF1-R antibody or antigen-binding fragment thereof. (Item 10) 9. The method of claim 8, wherein the antibody or antigen-binding fragment thereof is an endosialin (TEM-1) antibody or antigen-binding fragment thereof. (Item 11) 11. The method of any one of items 5 to 10, wherein the radioimmunoconjugate comprises a metal complex of a chelating moiety. (Item 12) 12. The method of claim 11, wherein the metal complex comprises a radionuclide. (Item 13) 13. The method of claim 12, wherein the radionuclide is an alpha emitter. (Item 14) The radionuclide is astatine-211( 211 At), Bismuth-212( 212 Bi), Bismuth-213( 213 Bi), Actinium-225( 225 Ac), radium-223( 223 Ra), lead-212( 212 Pb), Thorium-227( 227 Th), and terbium-149 ( 149 14. The method of claim 13, wherein the α-emitter is selected from the group consisting of: (Item 15) The radionuclide 225 Item 15. The method according to item 14, wherein Ac. (Item 16) The radioimmunoconjugate has the following structure: [ka] wherein B is the targeting moiety; 16. The method according to any one of items 5 to 15. (Item 17) 17. The method of any one of items 1 to 16, wherein the DDRi is a PARP inhibitor. (Item 18) 18. The method of claim 17, wherein the PARP inhibitor is a small molecule PARP inhibitor. (Item 19) 19. The method of item 18, wherein the small molecule PARP inhibitor is selected from the group consisting of niparib, niraparib, olaparib, talazoparib, pamiparib, rucaparib (camsylate), and veliparib, or an analog thereof. (Item 20) 20. The method of item 19, wherein the small molecule PARP inhibitor is olaparib or an analog thereof. (Item 21) 17. The method of any one of items 1 to 16, wherein the DDRi is an ATR inhibitor. (Item 22) 22. The method of item 21, wherein the ATR inhibitor is a small molecule ATR inhibitor. (Item 23) 23. The method of item 22, wherein the small molecule ATR inhibitor is selected from the group consisting of AZ20, AZD0156, AZD1390, AZD6738, BAY-1895344, EPT-46464, M3541, M4344, M6620 (formerly known as VE-922 or VX-970), NU6027, VE-821, or an analog thereof. (Item 24) 24. The method of item 23, wherein the ATR inhibitor is BAY-1895344 or an analog thereof. (Item 25) 17. The method of any one of items 1 to 16, wherein the DDRi is a WEE1 inhibitor, a Chk1 inhibitor, or a Chk2 inhibitor. (Item 26) 17. The method of any one of items 1 to 16, wherein the DDRi is a DNA protein kinase (DNA-PK) inhibitor. (Item 27) 27. The method of any one of items 1 to 26, wherein the mammal is a human. (Item 28) 28. The method of any one of items 1 to 27, wherein the mammal has been diagnosed with cancer. (Item 29) 29. The method of any one of items 1 to 28, wherein the cancer is 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, sarcoma, adrenocortical carcinoma, neuroendocrine carcinoma, Ewing's sarcoma, multiple myeloma, or acute myeloid leukemia. (Item 30) 30. The method of any one of items 1 to 29, wherein the mammal has at least one solid tumor. (Item 31) 31. The method of any one of items 1 to 30, wherein said administering provides a therapeutic effect. (Item 32) 32. The method of claim 31, wherein the therapeutic effect comprises a decrease in tumor volume, a stable tumor volume, or a decrease in the rate of tumor volume growth. (Item 33) 33. The method according to item 31 or 32, wherein the therapeutic effect comprises a reduction in the incidence of recurrence or metastasis. [Brief explanation of the drawings]

[0091] [Figure 1] Figure 1 is a schematic diagram showing the dosing schedule of [225Ac]-FPI-1434 and BAY-1895344 (ATR inhibitor) in two xenograft models: Colo-205 and A549. See Example 1.

[0092] [Figure 2] Figures 2A-2B show the relative tumor volumes of Colo-205 (Figure 2A) and A549 (Figure 2B) after treatment with BAY-1895344 alone and [225Ac]-FPI-1434 alone or in combination. See Example 1.

[0093] [Figure 3] 3 is a schematic diagram showing the dosing schedule of [225Ac]-FPI-1434 and BAY-1895344 in a Colo-205 (colorectal cancer) xenograft model. See Example 2.

[0094] [Figure 4] Figure 4 shows the relative tumor volume of Colo-205 after treatment with BAY-1895344 alone and [225Ac]-FPI-1434 alone or in combination. See Example 2.

[0095] [Figure 5] Figure 5 is a schematic diagram showing the dosing schedule of olaparib for treatment in two xenograft models: Colo-205 and A549. See Example 3.

[0096] [Figure 6] 6A-6B show the relative tumor volumes of Colo-205 (FIG. 6A) and A549 (FIG. 6B) after treatment with olaparib. See Example 3.

[0097] [Figure 7] 7 is a schematic diagram showing the dosing schedule of [225Ac]-FPI-1434 and olaparib for treatment in two xenograft models: Colo-205 and A549. See Example 4.

[0098] [Figure 8] Figures 8A-8B show the relative tumor volumes of Colo-205 (Figure 8A) and A549 (Figure 8B) after treatment with olaparib alone and [225Ac]-FPI-1434 alone or in combination. See Example 4.

[0099] [Figure 9] 9 is a schematic diagram showing the dosing schedule of [225Ac]-FPI-1434 and olaparib for treatment in the Colo-205 xenograft model. See Example 5.

[0100] [Figure 10]Figure 10 shows the relative tumor volume of Colo-205 after treatment with olaparib alone and [225Ac]-FPI-1434 alone or in combination. See Example 5.

[0101] [Figure 11] FIG. 11 is a schematic diagram showing the dosing schedule of [225Ac]-FPI-1434 and olaparib used in the multiple-dose in vivo study described in Example 6.

[0102] [Figure 12] FIG. 12 shows the relative tumor volumes of animals receiving the lower effective doses of [225Ac]-FPI-1434 (20 nCi) and olaparib (25 mg / kg) in the experiment described in Example 6.

[0103] [Figure 13-1] Figures 13A-13C show the relative tumor volumes of animals administered 20 nCi (Figure 13A), 50 nCi (Figure 13B), or 100 nCi (Figure 13C) of [225Ac]-FPI-1434 with olaparib (25 mg / kg or 50 mg / kg). [Figure 13-2] Figures 13A-13C show the relative tumor volumes of animals administered 20 nCi (Figure 13A), 50 nCi (Figure 13B), or 100 nCi (Figure 13C) of [225Ac]-FPI-1434 with olaparib (25 mg / kg or 50 mg / kg). DETAILED DESCRIPTION OF THE INVENTION

[0104] It should be understood that the drawings are not necessarily drawn to scale, and that objects within 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 devices, systems, and methods disclosed herein. Wherever possible, the same reference numbers will be 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.

[0105] The present disclosure relates to combination therapies for treating or ameliorating cancer using radioimmunoconjugates and DNA damage and repair inhibitors (DDRi). In some embodiments, the radioimmunoconjugates and / or DDRi are used at lower effective doses.

[0106] Radiolabeled targeting moieties (also known as radioimmunoconjugates) are designed to target proteins or receptors upregulated in disease states and / or specific to diseased cells (e.g., tumor cells) and deliver a radioactive payload to damage and kill the target cells. "Radioimmunotherapy" refers to this therapy when the targeting moiety comprises an antibody, typically a monoclonal antibody. Radioactive decay of the payload generates α, β, or γ particles or Auger electrons, which can cause direct effects on DNA (e.g., single- or double-stranded DNA breaks) or indirect effects such as bystander or crossfire effects.

[0107] Radioimmunoconjugates generally include a biological targeting moiety (e.g., an antibody or antigen-binding fragment thereof that specifically binds to a molecule expressed on or by a tumor, e.g., IGF-1R or TEm-1 / endosialin), a chelating moiety or a metal complex of a chelating moiety (e.g., containing a radioisotope), and a linker. Conjugates may be formed by appending a bifunctional chelate to a biological targeting molecule with minimal structural change while maintaining target affinity. Radioimmunoconjugates may be formed by radiolabeling such conjugates.

[0108] Bifunctional chelates structurally comprise chelate, linker and bridging group.When developing new bifunctional chelates, most efforts are focused on the chelate part of the molecule.Several examples of bifunctional chelates have been described, with various cyclic and acyclic structures conjugated to targeting moieties [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].

[0109] radioimmunoconjugate Radioimmunoconjugates suitable for use in accordance with the present disclosure generally have the structure of Formula Ia: ALB Formula Ia wherein A is a chelating moiety or a metal complex thereof; where B is a targeting moiety; In the formula, L is a linker.

[0110] In some embodiments, the radioimmunoconjugate has the following structure: [ka] where B is a targeting moiety. Includes.

[0111] targeting part Targeting moieties include molecules or any portion of a molecule that can bind to a given target. In some embodiments, the targeting moiety comprises a protein or polypeptide. In some embodiments, the targeting moiety is selected from the group consisting of an antibody or antigen-binding fragment thereof, a nanobody, an affibody, and a consensus sequence for a fibronectin type III domain (e.g., centilin or adnectin). In some embodiments, the moiety is both a targeting moiety and a therapeutic moiety. That is, the moiety can bind to a given target and also provides a therapeutic benefit.

[0112] antibody 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 each 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).

[0113] Antibodies described herein can 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 can 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.

[0114] 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 encompassed by 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" comprises 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.

[0115] 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 method described in Morrison, 1985, Science 229:1202, and humanized antibodies can be produced, for example, by the method described in U.S. Pat. No. 6,180,370.

[0116] Additional antibodies described herein are bispecific and multivalent antibodies, for example, as described in Segal et al., J. Immunol. Methods 248:1-6 (2001); and Tutt et al., J. Immunol. 147:60 (1991).

[0117] Insulin-like growth factor 1 (IGF-1R) antibodies The insulin-like growth factor 1 receptor (IGF-1R) is a transmembrane protein found on the surface of human cells activated by insulin-like growth factors 1 (IGF-1) and 2 (IGF-2). In some embodiments, the radioimmunoconjugate contains an antibody against the insulin-like growth factor 1 receptor (IGF-1R). Although not a typical oncogene, IGF-1R promotes cancer initiation and progression and plays an important role in mitogenic transformation and maintenance of the transformed phenotype. IGF-1R has been implicated in the development of several 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 the 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.

[0118] Despite a wealth of data implicating the role of IGF-1R in cancer, therapeutics targeting IGF-1R have yet to demonstrate significant disease impact. Much speculation has been made about this lack of efficacy, including the inability 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 overexpressing the IGF-1R by exploiting the ability of IGF-1R to undergo antibody-induced internalization and lysosomal degradation to deliver targeted radioisotopes into 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 potential for cell-killing 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 due to direct DNA bombardment and destruction by alpha particles can occur in the targeted cell or within a radius of two or three untargeted cells for a given alpha particle decay. In addition to their potentially very high antitumor efficacy, IGF-1R-targeted radioimmunoconjugates may not develop mechanistic resistance because they do not rely on blocking the receptor-binding ligand to inhibit the oncological process, as is required for therapeutic antibodies.

[0119] Several IGF-1R antibodies, including figitumumab, cizutumumab, ganitumab, AVE1642 (also known as humanized EM164 and huEM164), BIIB002, lobatumumab, and teprotumumab, have been developed and investigated for the treatment of various types of cancer. After binding to IGF-1R, these antibodies are internalized within 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 exposure of normal tissues to toxic agents. The CDRs of the light chain variable region of AVE1642 have the following sequences: SEQ ID NO: 1 (CDR-L1) RSSQSIVHSNVNTYLE Sequence number 2 (CDR-L2) KVSNRFS SEQ ID NO: 3 (CDR-L3) FQGSHVPPT The light chain variable region of AVE1642 has the following sequence: SEQ ID NO:4 DVVMTQTPLSLPVSLGDPASISCRSSQSIVHSNVNTYLEWYLQKPGQSPRLLIYKVSNRF SGVPDRFSGSGAGTDFTLRISRVEAEDLGIYYCFQGSHVPPTFGGGTKLEIKRTVAAPSV FIFPPSDEQLKSGTASVVCLLNNFYPREAK The CDRs of the heavy chain variable region of AVE1642 have the following sequences: SEQ ID NO: 5 (CDR-H1)SYWMH SEQ ID NO: 6 (CDR-H2) GEINPSNGRTNY NQKFQG SEQ ID NO: 7 (CDR-H3) GRPDYYGSSKWY FDV The heavy chain variable region of AVE1642 has the following sequence: SEQ ID NO:8 QVQLVQSGAEVVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGEINPSNGRTNY NQKFQGKATLTVDKSSSTAYMQLSSLTSEDSAVYYFARGRPDYYGSSKWYFDVWGQGTTV TVSSASTKGPSVFPLAPSSKSTSGGTAALG

[0120] 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.

[0121] 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).

[0122] In some embodiments, the endosialin antibody or antigen-binding fragment thereof recognizes an epitope having the amino acid sequence of SRDHQIPVIAAN (SEQ ID NO: 9).

[0123] In some embodiments, the heavy chain variable region of an endosialin antibody or antibody-binding fragment thereof comprises complementarity-determining regions (CDRs) having the following sequences: CDR-H1: GYGVN (SEQ ID NO: 10) or GFSLTGYGVN (SEQ ID NO: 11) CDR-H2: MIWVDGSTDYNSALKS (SEQ ID NO: 12) CDR-H3: GGYGAMDY (SEQ ID NO: 13)

[0124] In some embodiments, the light chain variable region of an endosialin antibody or antibody-binding fragment thereof comprises complementarity-determining regions (CDRs) having the following sequences: CDR-L1: HASQNINVWLT (SEQ ID NO: 14) CDR-L2: KASNLHT (SEQ ID NO: 15) CDR-L3: QQGQSYPWT (SEQ ID NO: 16)

[0125] In some embodiments, the endosialin antibody or antigen-binding fragment thereof is a humanized antibody.

[0126] In some embodiments, the heavy chain variable region of the endosialin antibody or antigen-binding fragment thereof comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 17, 18, 19, or 20: Humanized VH1: QVQLQESGPGLVKPSETLSLTCTVSGFSLTGYGVNWIRQPPGKGLEWIGMIWVDGSTDYN SALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGGYGAMDYWGQGTLVTVSS (SEQ ID NO: 17) Humanized VH2: QVQLQESGPGLVKPSETLSLTCTVSGFSLTGYGVNWIRQPPEKGLEWIGMIWVDGSTDYN SALKSRVNISVDTSKNQFSLKLSSVTAADTAVYYCARGGYGAMDYWGQGTLVTVSS (SEQ ID NO: 18) Humanized VH3: QLQLQESGPGLVKPSETLSLTCTVSGFSLTGYGVNWIRQPPGKGLEWIGMIWVDGSTDYN SALKSRVTISVDKSKNQFSLKLSSVTAADTAVYYCARGGYGAMDYWGQGTLVTVSS (SEQ ID NO: 19) Humanized VH4: QLQLQESGPGLVKPSETLSLTCTVSGFSLTGYGVNWIRQPPEKGLEWIGMIWVDGSTDYN SALKSRVNISVDKSKNQFSLKLSSVTAADTAVYYCARGGYGAMDYWGQGTLVTVSS (SEQ ID NO: 20)

[0127] In some embodiments, the light chain variable region of the endosialin antibody or antigen-binding fragment thereof comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 21, 22, 23, or 24: Humanized VL1: DIQMTQSPSSVSASVGDRVTITCHASQNINVWLTWYQQKPGKAPKLLIYKASNLHTGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQGQSYPWTFGGGTKLEIK (SEQ ID NO: 21) Humanized VL2: DIQMTQSPSTLSASVGDRVTITCHASQNINVWLTWYQQKPGKAPKLLIYKASNLHTGVPS RFSGSGSGTEFTLTISSLQPDDFATYYCQQGQSYPWTFGGGTKLEIK (SEQ ID NO: 22) Humanized VL3: DIQMTQSPSSLSASVGDRVTITCHASQNINVWLTWYQQKPGKAPKLLIYKASNLHTGVPS RFSGSGSGTDFTFTISSLQPEDIATYYCQQGQSYPWTFGGGTKLEIK (SEQ ID NO: 23) Humanized VL4: DIQMTQSPSSLSASVGDRVTITCHASQNINVWLTWYQQKPEKAPKSLIYKASNLHTGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQGQSYPWTFGGGTKLEIK (SEQ ID NO: 24)

[0128] 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. Nanobodies can be biologically engineered by site-directed mutagenesis or mutagenic screening (e.g., phage display, yeast display, bacterial display, mRNA display, ribosome display).

[0129] Affibody Affibodies are polypeptides or proteins engineered to bind to specific antigens. 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 aureus protein A. Affibodies can be engineered variants of the Z domain, a B domain with low affinity for 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).

[0130] 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.

[0131] Fibronectin type III domain The fibronectin type III domain is an evolutionarily conserved protein domain found in diverse extracellular proteins. It has been used as a molecular scaffold to generate molecules capable of selectively binding to specific antigens. Fibronectin type III domain (FN3) variants engineered for selective binding are sometimes called monobodies. FN3 domains can be biologically engineered by site-directed mutagenesis or mutagenic screening (e.g., CIS-display, phage display, yeast display, bacterial display, mRNA display, and ribosome display).

[0132] Modified Polypeptides The polypeptides used according to the present disclosure may have modified amino acid sequences. The modified polypeptides may be substantially identical to the corresponding reference polypeptides (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 (e.g., by at least 5%, 10%, 20%, 25%, 35%, 50%, 60%, 70%, 75%, 80%, 90%, or 95%), have no effect, or increase (e.g., by at least 5%, 10%, 25%, 50%, 100%, 200%, 500%, or 1000%) 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.

[0133] Modifications include those that result from natural processes, such as post-translational processing, or from chemical modification techniques known in the art. Modifications can occur anywhere in a polypeptide, including the polypeptide backbone, amino acid side chains, and the amino or carboxy termini. The same type of modification can be present in the same or varying 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 an acetomidomethyl (Acm) group, 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.

[0134] Modified polypeptides can also include amino acid insertions, deletions, or conservative or non-conservative substitutions (e.g., D-amino acids, desamino 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 terminus of a polypeptide can facilitate conjugation of these polypeptides, for example, by disulfide bonds. For example, polypeptides can be modified to contain 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 with another of the same general type or group) or non-conservative (i.e., a residue is replaced with an amino acid of a different type). Furthermore, 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).

[0135] 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 with an acetylaminomethyl group attached to the sulfur atom of cysteine, pegylated amino acids, amino acids with the formula NH2(CH2), n These include COOH omega amino acids (where n is 2-6), neutral nonpolar 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 nonpolar, cysteic acid is acidic, and ornithine is basic. Proline can be substituted with hydroxyproline and retain its conformational properties.

[0136] 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 undesired changes, other types of substitutions designated "exemplary substitutions" in Table 1 or further described herein in relation to amino acid classes are introduced and the products screened. [Table 1]

[0137] 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 the bulk of the side chains.

[0138] Chelating moieties and their metal complexes Chelate part Examples of suitable chelating moieties include, but are not limited to, 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-tetrapropionic acid), DOAM-acetate (2-(4,7,10-tris(2-amino-2-oxoethyl)-1,4,7,10-tetraazacyclododecane)- ... cyclododecan-1-yl)acetic acid), DOTA-GA anhydride (2,2',2"-(10-(2,6-dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid, DOTP (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylenephosphonic acid)), DOTMP (1,4,6,10-tetraazacyclodecane-1,4,7,10-tetramethylenephosphonic 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) 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 (tetraazacyclododecanedimethanephosphonic acid), HP-DO3A (hydroxypropyltetraazacyclododecanetriacetic acid), EDTA (ethylenediaminetetraacetic acid), deferoxamine, DTPA (diethylenetriaminepentaacetic acid), DTPA-BMA (diethylenetriaminepentaacetic acid-bismethylamide), HOPO (octadentate hydroxypyridinone), or porphyrin.

[0139] In some embodiments, the radioimmunoconjugate comprises a metal complex of a chelating moiety. For example, chelating groups may be used in combination with metals, such as manganese, iron, and gadolinium, and isotopes (e.g., isotopes in the general energy range of 60-4000 keV), such as any of the radioisotopes and radionuclides discussed herein. 2 , 2.2 ,

[0140] In some embodiments, chelating moieties are useful as detection agents, and radioimmunoconjugates containing such detectable chelating moieties can therefore be used as diagnostic or theranostic agents.

[0141] Radioisotopes and Radionuclides In some embodiments, the metal complex comprises a radionuclide. Examples of suitable radioisotopes and radionuclides include, but are not limited to: 3 H, 14 C. 15 N, 18 F, 35 S, 47 Sc, 55 Co, 60 Cu, 61 Cu, 62 Cu, 64 Cu, 66 Ga, 67 Ga, 67 Cu, 68 Ga, 75 Br, 76 Br, 77 Br, 82 Rb, 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, 117m Sn, 186 Re, 188 Re, 198 Au, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Pb, 212 Bi, 213 Bi,223 Ra, 225 Ac, 227 Th, and 229 Th is an example.

[0142] In some embodiments, the radionuclide is an alpha emitter, e.g., astatine-211 ( 211 At), Bismuth-212( 212 Bi), Bismuth-213( 213 Bi), Actinium-225( 225 Ac), radium-223( 223 Ra), lead-212( 212 Pb), Thorium-227( 227 Th), or terbium-149 ( 149 Tb).

[0143] Linker In some embodiments, the linker is the moiety of Formula Ib that does not include A and B, as shown in the structure of Formula Ib: AL 1 -(L 2 ) n -B Formula Ib (A and B are as defined in formula Ia). Thus, in some embodiments, the linker is -L 1 -(L 2 ) n - and In the formula, L 1 is optionally substituted C1-C6 alkyl, substituted C1-C6 heteroalkyl, substituted aryl or heteroaryl; n is 1 to 5; Each L 2 independently have the following structure: (-X 1 -L 3 -Z 1 -) Formula II In the formula, X 1 is C=O(NR 1 ), C=S(NR 1 ), O-C=O(NR 1 ), NR1 C=O(O), NR 1 C=O(NR 1 ), -CH2PhC=O(NR 1 ), -CH2Ph(NH)C=S(NR 1 ), O, or NR 1 and each R 1 are independently H or optionally substituted C1-C6 alkyl or optionally substituted C1-C6 heteroalkyl, substituted aryl or heteroaryl, wherein C1-C6 alkyl can be substituted with oxo (=O), heteroaryl, or combinations thereof; L 3 is C1-C, optionally substituted 50 Alkyl or optionally substituted C1-C 50 Heteroalkyl or C5-C 20 Polyethylene glycol; Z 1 are CH2, C=O, C=S, OC=O, NR 1 C=O, NR 1 and R 1 is hydrogen or optionally substituted C1-C6 alkyl, pyrrolidine-2,5-dione.

[0144] crosslinking group In some embodiments, the radioimmunoconjugate comprises a crosslinking group instead of, or in addition to, a targeting moiety or a therapeutic moiety (eg, B of Formula I comprises a crosslinking group).

[0145] A "bridging group" is a reactive group capable of covalently linking two or more molecules. Bridging groups can be used to attach linkers and chelating moieties to therapeutic or targeting moieties. Bridging groups can also be used to attach linkers and chelating moieties to targets in vivo. In some embodiments, the bridging group is an amino-reactive, methionine-reactive, or thiol-reactive bridging group, or sortase-mediated coupling. In some embodiments, the amino-reactive or thiol-reactive bridging group comprises an activated ester, such as a hydroxysuccinimide ester, a 2,3,5,6-tetrafluorophenol ester, a 4-nitrophenol ester, or an imidate, an anhydride, a thiol, a disulfide, a maleimide, an azide, an alkyne, a strained alkyne, a strained alkene, a halogen, a sulfonate, a haloacetyl, an amine, a hydrazide, a diazirine, a phosphine, a tetrazine, an isothiocyanate, or an oxaziridine. In some embodiments, the sortase recognition sequence may comprise a terminal glycine-glycine-glycine (GGG) and / or an LPTXG amino acid sequence, where X is any amino acid. One of skill in the art will understand that the use of cross-linking groups is not limited to the particular constructs disclosed herein, but rather may include other known cross-linking groups.

[0146] DNA damage and repair inhibitors (DDRi) In various embodiments, a DNA damage and repair inhibitor (DDRi) is co-administered with the radioimmunoconjugate. DNA repair involves multiple molecular pathways that repair DNA single-strand breaks (e.g., the PARP pathway) and double-strand breaks (e.g., other genes such as BRCA and ATR / ATM). PARP inhibition (PARPi) results in failure of single-strand break repair, leading to further double-strand breaks. Available PARP inhibitors act by both PARP enzyme inhibition and DNA trapping. Tumor cells with BRCA and / or PTEN mutations are sensitive to PARPi. ATR inhibition (ATRi) results in failure of double-strand break repair, and the accumulation of double-strand breaks leads to cell death. These inhibitors act by preventing homologous recombination and non-homologous end-joining.

[0147] The present disclosure relates to combination therapy with a radioimmunoconjugate and a DNA damage and repair inhibitor. This type of combination therapy has been found to provide unexpected improvements in cancer treatment, particularly in cancers not expected to be responsive to DDRi. In some embodiments, the DDRi is a PARP inhibitor (PARPi). In some embodiments, the PARPi is selected from the group including niparib, niraparib, olaparib, pamiparib, rucaparib (camsylate), talazoparib, veliparib, or analogs thereof. In some embodiments, the DDRi is an ATM / ATR inhibitor. In some embodiments, the ATM / ATR inhibitor is selected from the group including AZ20, AZD0156, AZD1390, AZD6738, BAY-1895344, EPT-46464, M3541, M4344, M6620 (formerly known as VE-922 or VX-970), NU6027, VE-821, or an analog thereof. In some embodiments, the PARPi is adavosertib, AZD2811, or an analog thereof. In some embodiments, the DDRi is a WEE1 inhibitor, a Chk1 inhibitor, or a Chk2 inhibitor. In some embodiments, the DDRi is a DNA-dependent protein kinase (DNA-PK) inhibitor. Non-limiting examples of DNA-PK inhibitors include AZD7648, KU-0060648, NU7026, NU7441 (KU-57788), PI-103, PIK-75 HCI, PP121, SF2523, and analogs thereof.

[0148] Subject In some disclosed methods, a therapy (e.g., comprising a therapeutic agent) is administered to a subject. In some embodiments, the subject is a mammal, e.g., a human.

[0149] In some embodiments, the subject has been or is currently receiving another treatment. For example, in some embodiments, the subject has been or is currently receiving a radioimmunoconjugate. In some embodiments, the subject has been or is currently receiving a DNA damage and repair inhibitor (DDRi).

[0150] 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.

[0151] In some embodiments, the cancer is a solid tumor.

[0152] In some embodiments, the solid tumor cancer is breast cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, head and neck cancer, prostate cancer, colorectal cancer, sarcoma, adrenocortical carcinoma, neuroendocrine carcinoma, Ewing's sarcoma, multiple myeloma, or acute myeloid leukemia.

[0153] In some embodiments, the cancer is a non-solid (e.g., liquid (e.g., blood)) cancer.

[0154] Administration and Dosage Effective doses and lower effective doses The present disclosure provides a combination therapy, in which the amount of each therapeutic agent may or may not be therapeutically effective by itself.For example, provided is a method comprising administering a first treatment and a second treatment together in an amount that is effective for treating or improving a disorder, such as cancer.In some embodiments, at least one of the first and second treatments is administered to a subject at a lower effective dose.In some embodiments, both the first and second treatments are administered at a lower effective dose.

[0155] In some embodiments, the first treatment comprises a radioimmunoconjugate and the second treatment comprises a DNA damage response inhibitor (DDRi).

[0156] In some embodiments, the first treatment comprises a DDRi and the second treatment comprises a radioimmunoconjugate.

[0157] 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" typically varies depending on the therapeutic agent. For known therapeutic agents, the relevant therapeutically effective amount is known to or can be easily determined by those skilled in the art.

[0158] For example, in the treatment of cancer, a drug or compound that reduces, prevents, delays, inhibits, or suppresses any symptoms of the 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 treatment for the 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 recovery is accelerated. For example, a treatment can be therapeutically effective if it causes cancer to regress or slows the growth of cancer.

[0159] Effective dosing regimens for these uses (e.g., the amount of each therapeutic agent, the 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 administered to a mammal (e.g., a human) can be determined by one of ordinary skill in the art, taking into account individual differences in the mammal's age, weight, and condition. 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 the therapeutic effect of the unconjugated drug (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 can also be determined empirically by one of ordinary skill in the art. Accordingly, lower effective doses may also be determined by one of ordinary skill in the art.

[0160] Single or multiple administrations of a composition (e.g., a pharmaceutical composition comprising a therapeutic agent) can be carried out with dose levels and patterns 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 can be monitored throughout the course of treatment according to methods commonly practiced by clinicians or methods described herein.

[0161] In the disclosed combination therapy methods, the first and second therapies may be administered to a subject sequentially or simultaneously. For example, a first composition containing a first therapeutic agent and a second composition containing a second therapeutic agent may be administered to a subject sequentially or simultaneously. Alternatively, or in addition, a composition containing a combination of a first therapeutic agent and a second therapeutic agent may be administered to a subject.

[0162] 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.

[0163] In some embodiments, the DDRi is administered in a single dose. In some embodiments, the DDRi is administered multiple times (e.g., at least twice, at least three times, etc.). In some embodiments, the DDRi 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 DDRi is administered multiple times, the dose of each administration may be the same or different. For example, the DDRi may be administered in an initial dose, and the dosage of the subsequent DDRi may be higher or lower than the initial dose.

[0164] In some embodiments, the first dose of DDRi is administered simultaneously with the first dose of radioimmunoconjugate. In some embodiments, the first dose of DDRi is administered before the first dose of radioimmunoconjugate. In some embodiments, the first dose of DDRi is administered after the first dose of radioimmunoconjugate. In some embodiments, a subsequent dose of DDRi is administered.

[0165] In some embodiments, the radioimmunoconjugate (or composition thereof) and the DDRi (or composition thereof) are administered within 28 days (eg, within 14, 7, 6, 5, 4, 3, 2, or 1 day) of each other.

[0166] In some embodiments, the radioimmunoconjugate (or composition thereof) and the DDRi (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 DDRi is administered simultaneously with the radioimmunoconjugate. In various embodiments, the DDRi is administered multiple times following the first administration of the radioimmunoconjugate.

[0167] 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 can 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 a disclosed conjugate or 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.

[0168] Pharmaceutical compositions containing one or more drugs (e.g., radioimmunoconjugates and / or DDRi) 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 appropriate formulation. Examples of suitable formulations can be found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, PA, 17th ed., 1985. For a brief review of methods for drug delivery, see, for example, Langer (Science 249:1527-1533, 1990).

[0169] formulation Pharmaceutical compositions can be formulated for local administration, such as by parenteral, intranasal, topical, oral, or transdermal means, for prophylactic and / or therapeutic treatment. Pharmaceutical compositions can be administered parenterally (e.g., by intravenous, intramuscular, or subcutaneous injection), or by oral ingestion, or by local application to an area affected by a vascular or cancerous condition or by intraarticular injection. Additional exemplary routes of administration include intravascular, intraarterial, intratumoral, intraperitoneal, intracerebroventricular, intraepithelial, as well as nasal, ocular, intrascleral, intraorbital, rectal, topical, or aerosol inhalation administration. Sustained-release administration, such as by depot injection or erodible implants or components, is also specifically contemplated. 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, for parenteral administration. The composition may contain pharmaceutically 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.

[0170] The compositions may be sterilized, for example, by conventional sterilization techniques or sterile filtered. Aqueous solutions may be packaged ready for 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 to 11, more preferably 5 to 9 or 6 to 8, and most preferably 6 to 7, e.g., 6 to 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-described agents, such as a sealed package of tablets or capsules. In some embodiments, the solid form of the composition is packaged in a flexible volume container, such as a squeezable tube designed for topical creams or ointments. Other drugs

[0171] In some embodiments, the disclosed methods further comprise administration of an antiproliferative agent, a radiosensitizer, or an immunosuppressant or immunomodulatory agent.

[0172] "Antiproliferative" or "antiproliferative agent" are used interchangeably herein and refer to anticancer agents, including those listed in Table 2, any of which can 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.

[0173] "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 a radioimmunoconjugate.

[0174] As used herein, the term "radiosensitizer" refers to any agent that enhances the sensitivity of cancer cells to radiation therapy. Radiation sensitizers may 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 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6] [Table 2-7] [Table 2-8] [Table 2-9] [Table 2-10] [Example]

[0175] Example 1. In vivo synergy test: 225 Ac]-FPI-1434+BAY-1895344-Administration Schedule 1 In vivo synergy testing was performed using 225 Ac]-FPI-1434 and BAY-1895344. 225 Ac]-FPI-1434 was administered as a single agent at a suboptimal dose for tumor regression to Colo-205 (colorectal cancer) xenograft models (50 nCi) and A549 (NSCLC) xenograft models (200 nCi). The combination group (n=5) received BAY-1895344 at a dose of 20 mg / kg (intraperitoneally) for 28 days, with 2 days on and 5 days off. The first dose of BAY-1895344 was [ 225 Ac]-FPI-1434 was administered 24 hours after administration of FPI-1434. See Figure 1.

[0176] Relative tumor volume was assessed up to 28 days after administration. In both the Colo-205 and A549 xenograft models, BAY-1895344 alone did not show any reduction in tumor volume over time compared to vehicle controls. In both models, [ 225 Treatment with [Ac]-FPI-1434 alone showed a significant reduction in tumor volume compared to vehicle control and treatment with BAY-1895344 alone. 225 When [Ac]-FPI-1434 and BAY-1895344 were administered as a combination therapy using the dosing schedule in Figure 1, no additional benefit was observed compared to either treatment administered alone (see Figures 2a and 2b).

[0177] Example 2. In vivo synergy study: FPI-1434 + BAY-1895344 - Dosing schedule 2 In vivo synergy studies were performed using a second dosing schedule [ 225A study was conducted with [Ac]-FPI-1434 and BAY-1895344. FPI-1434 was administered as a single agent at a suboptimal dose (50 nCi) for tumor regression in a Colo-205 (colorectal cancer) xenograft model. In the combination group (n=5), BAY-1895344 was administered three times weekly, starting at a dose of 40 mg / kg. Transient somnolence in the animals was observed at higher doses, so the dose was gradually reduced to 20 mg / kg. The first dose of BAY-1895344 was administered 24 hours after FPI-1434 administration. See Figure 3.

[0178] Relative tumor volume was assessed up to 28 days after administration. 225 Both Ac]-FPI-1434 and BAY-1895344 alone showed some reduction in tumor volume over time, and [ 225 Ac]-FPI-1434 was more effective in reducing tumor volume than BAY-1895344 alone. 225 When [Ac]-FPI-1434 and BAY-1895344 were administered as a combination therapy using the dosing schedule in Figure 3, animals exhibited significantly lower tumor volumes compared to either treatment alone. See Figure 4.

[0179] Example 3. In vivo efficacy of olaparib as a single agent An in vivo efficacy study was conducted using olaparib alone as a single agent. Olaparib was administered at different doses as a single agent to Colo-205 (colorectal cancer) xenograft models and A549 (NSCLC) xenograft models. Animal groups (n=5) were administered olaparib (intraperitoneally) at doses ranging from 10 to 100 mg / kg, with a 5-day on-2-day off schedule. See Figure 5.

[0180] Olaparib demonstrated modest efficacy as a single agent in both models (Figures 6A and 6B). No dose response was observed in either model, and immunohistochemical analysis of y-H2AX foci did not demonstrate increased DSB formation after olaparib treatment.

[0181] Example 4. In Vivo Synergy Study: FPI-1434 + Olaparib - Dosing Schedule 1 In vivo synergy testing was performed using 225 Ac]-FPI-1434 and olaparib. 225 Ac]-FPI-1434 was administered as a single agent at suboptimal doses for tumor regression in Colo-205 (colorectal cancer) xenograft models (50 nCi) and A549 (NSCLC) xenograft models (200 nCi). In the combination group (n=5), olaparib was administered intraperitoneally at a dose of 50 mg / kg for 28 days, 5 days on, 2 days off. 225 Animals were pre-treated with olaparib three times prior to administration of [Ac]-FPI-1434. See Figure 7.

[0182] Relative tumor volume was assessed up to 28 days after administration. In the Colo-205 xenograft model, 225 Both Ac]-FPI-1434 and olaparib alone showed some reduction in tumor volume over time, and [ 225 Ac]-FPI-1434 was more effective in reducing tumor volume than olaparib alone. 225 When [Ac]-FPI-1434 and olaparib were administered as a combination therapy using the dosing schedule in Figure 7, no additional benefit was observed compared to either treatment administered alone. See Figure 8a.

[0183] In the A549 xenograft model, treatment with olaparib alone did not result in a clear difference in tumor volume over time compared to untreated animals. 225 Ac]-FPI-1434 significantly reduced tumor volume compared to vehicle- and olaparib-treated animals. 225 When [Ac]-FPI-1434 and olaparib were administered as a combination therapy using the dosing schedule in Figure 7, no additional benefit was observed compared to either treatment administered alone. See Figure 8b.

[0184] Example 5. In Vivo Synergy Study: FPI-1434 + Olaparib - Dosing Schedule 2 In vivo synergy studies were performed using a second dosing schedule [ 225 A study was conducted with [Ac]-FPI-1434 and olaparib. FPI-1434 was administered as a single agent at a suboptimal dose (50 nCi) for tumor regression to a Colo-205 (colorectal cancer) xenograft model. The combination group (n=5) received olaparib (intraperitoneally) at a dose of 50 mg / kg for 28 days, with 5 days on and 2 days off. The first dose of olaparib was administered 24 hours after FPI-1434 administration. See Figure 9.

[0185] Relative tumor volume was assessed up to 28 days after administration. 225 Both Ac]-FPI-1434 and olaparib alone showed significant reductions in tumor volume over time, [ 225 Ac]-FPI-1434 was more effective in reducing tumor volume than olaparib alone. 225 When [Ac]-FPI-1434 and olaparib were administered as a combination therapy using the dosing schedule in Figure 9, animals exhibited significantly lower tumor volumes compared to either treatment alone. See Figure 10.

[0186] Example 6. In vivo synergy study: FPI-1434 + olaparib combination therapy at multiple dose levels [ 225 In vivo studies were conducted in a Colo-205 (colorectal cancer) xenograft model using [Ac]-FPI-1434 and olaparib. 225 Ac]-FPI-1434 was administered at doses of 20 nCi, 50 nCi, or 100 nCi. In the combination group (n=5), animals received [ 225 Olaparib (intraperitoneal) administration was initiated 24 hours after Ac]-FPI-1434 administration, and [ 225 Ac]-FPI-1434 administration ("Day 0") through Day 30, doses of 25 or 50 mg / kg were administered according to a 5-day on, 2-day off schedule. See Figure 11.

[0187] Relative tumor volumes were assessed at various time points of administration. Figure 12 shows the relative tumor volumes at the lower effective doses. 225 Results are shown using [Ac]-FPI-1434 (20 nCi) and olaparib (25 mg / kg). 225 While no or limited therapeutic effect was observed with either [Ac]-FPI-1434 or olaparib alone, animals receiving the combination therapy exhibited significantly lower tumor volumes compared to animals receiving either treatment alone. Figures 13A-13C show the effect of 20 nCi (Figure 13A), 50 nCi (Figure 13B), or 100 nCi (Figure 13C) of [Ac]-FPI-1434 on tumor volume. 225 Figure 13 shows the results using [Ac]-FPI-1434 and olaparib (25 mg / kg or 50 mg / kg). The strongest combination effect was observed at the lowest single-agent dose. See Figure 13A.

[0188] Therefore, in this embodiment, 225 We demonstrate that the combination of Ac]-FPI-1434 and olaparib resulted in synergistic effects at lower effective doses of each.

[0189] Equivalents / Alternative Embodiments 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

[Claim 1] The invention described in the specification.

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