antibody-drug conjugates

Abstract

The present disclosure provides antibody-drug conjugates (ADCs) comprising an antibody, an antigen-binding fragment thereof, or a cell-penetrating fragment thereof, a linker, and a payload moiety. Compositions comprising the ADCs and methods of using the ADCs are also provided.

Description

[Technical Field]

[0001] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with government support under R35 CA197574 awarded by the National Institutes of Health. The government has certain rights in this invention.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 477,364, filed December 27, 2022, U.S. Provisional Patent Application No. 63 / 513,065, filed July 11, 2023, U.S. Provisional Patent Application No. 63 / 580,921, filed September 6, 2023, and U.S. Provisional Patent Application No. 63 / 585,915, filed September 27, 2023, the contents of which are incorporated herein by reference in their entirety for all purposes. [Background technology]

[0003] Antibody-drug conjugates (ADCs) are a rapidly growing class of targeted therapeutics and represent a promising new approach to improving drug selectivity and cytotoxic activity. These therapeutic agents consist of an antibody (or antibody fragment) covalently attached to a payload drug to form an immunoconjugate. The antibody directs the ADC to bind to target cells. Once inside the cell, the ADC can localize in and around the cell nucleus and liberate its payload, thereby treating or killing the cell.

[0004] The monoclonal 3E10 antibody ("3E10") has been developed as a molecular delivery vehicle for transporting various biologically important molecules to target cells, and 3E10 has been shown to preferentially target cancer cells. 3E10 penetrates cells via an Fc-independent mechanism, which involves the presence of the nucleoside transporter ENT2, as evidenced by the ability of Fc-depleted 3E10 fragments to penetrate cells (Weisbart et al., Scientific Reports volume 5, Article number: 12022 (2015) (Non-Patent Document 1); Zack et al., J. Immunol. 157, 2082-2088 (1996) (Non-Patent Document 2); Hansen et al., J. Biol. Chem. 282, 20790-20793 (2007) (Non-Patent Document 3)). The 3E10 single-chain variable fragment, 3E10 scFv, has previously been shown to be able to penetrate live cells and nuclei, with uptake efficiency reduced in ENT2-deficient cells (Hansen, et al., J. Biol. Chem. 282, 20790-20793 (2007) (Non-Patent Document 3)).

[0005] 3E10 has not demonstrated cytotoxicity in vitro or in vivo in studies to date. In contrast, some antibodies that penetrate live cells often exhibit toxicity and damage, which can trigger the pathological symptoms of the autoimmune diseases in which they are found. The 3E10 antibody is an ideal molecular delivery vehicle due to its specific nuclear localization, lack of toxicity, and efficiency in penetrating live cells with successful delivery of therapeutic cargo proteins in vitro and in vivo.

[0006] Currently, most ADCs are designed to target specific cancer types with their chemotherapy payloads based on cell surface antigen expression. For example, ADCs designed to combat breast cancer often target HER2, a cell surface receptor frequently expressed on breast cancer cells. New ADCs that improve drug delivery to target cells are needed, including new ADCs with the ability to deliver drugs to multiple cancer cell types. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Weisbart et al.,Scientific Reports volume 5,Article number:12022(2015) [Non-patent document 2] Zack et al.,J.Immunol.157,2082-2088(1996) [Non-patent document 3] Hansen et al.,J.Biol.Chem.282,20790-20793(2007) Summary of the Invention

[0008] In embodiments, the present disclosure provides a compound of formula (I): A-(LP r ) q Formula (I) wherein the formula (I) is: A is an antibody, an antigen-binding fragment thereof, or a cell-penetrating fragment thereof, comprising a heavy chain variable region (VH) comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 58, CDR2 comprising the amino acid sequence of SEQ ID NO: 59, and CDR3 comprising SEQ ID NO: 60, and a light chain variable region (VL) comprising CDR1 comprising the amino acid sequence of SEQ ID NO: 61, CDR2 comprising the amino acid sequence of SEQ ID NO: 62, and CDR3 comprising the amino acid sequence of SEQ ID NO: 63; L is a linker; P is the payload portion; r is an integer from 1 to 4; q is an integer from 1 to 16.

[0009] In some embodiments, the linker L is an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted arylene, an optionally substituted cycloalkylene, an optionally substituted heteroalkylene, an optionally substituted heteroarylene, an optionally substituted heterocycloalkylene, -NR a- (Linker number 1), -N=CR a- (Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -S(O)-(Linker No. 5), -S(O) 2- (Linker number 6), -OP(O)OR a- (Linker number 7), -OP(O)OR a O-(Linker No. 8), -P(O)OR a O-(Linker No. 9), -O-(Linker No. 10), -CR b 2- -(Linker number 11), -[(CR b 2) 1-12 O] 1-50 -(Linker No. 12), -C(O)-(Linker No. 13), -C(S)-(Linker No. 14), -C(=N-OH)-(Linker No. 15), -C(NR a )-(Linker No. 16), -C(NH2Cl)-(Linker No. 17), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)S-(Linker No. 20), -SC(O)-(Linker No. 21), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -C(O)NR a SO2- (linker number 24), -SO2NR a C(O)-(Linker No. 25), -OC(O)O-(Linker No. 26), -OC(O)S-(Linker No. 27), -SC(O)O-(Linker No. 28), -OC(O)NR a- (Linker No. 29), -NR a C(O)O- (Linker No. 30), -SC(O)NR a -(Linker No. 31), -NR aC(O)S- (Linker No. 32), -S(O) t N(R a )-(wherein t is 1 or 2) (Linker No. 33), -N(R a )S(O) t- (wherein t is 1 or 2) (Linker No. 34), and -X AA - (Linker No. 35); Each R a is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R b are independently hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -C(O)NR a 2. -CO2R a , -NR a 2, selected from optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Two independent R b group or R aand R b can be joined together to form an optionally substituted ring; and -X AA - is an amino acid sequence containing 1 to 6 amino acid moieties.

[0010] In some embodiments, -X AA is independently selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Val), citrulline (Cit), and homocitrulline (HoCit). In some embodiments, the linker L is an optionally substituted C1-C 18 Alkylene (Linker No. 36), -C≡C- (Linker No. 339), -CR a =CR a -(Linker Number 38), optionally substituted 6- to 14-membered arylene (Linker Number 39), optionally substituted C3-C 20 Cycloalkylene (Linker No. 40), -[CHO] 1-18 -(Linker No. 41), -[CH2CH2O] 1-18 -(Linker No. 42), -[CH2CH2CH2O] 1-18 -(Linker Number 43), optionally substituted 5- to 18-membered heteroarylene (Linker Number 44), optionally substituted 3- to 20-membered heterocycloalkylene (Linker Number 45), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -S(O)-(Linker No. 5), -S(O)2-(Linker No. 6), -OP(O)OR a -(Linker number 7), -OP(O)ORa O-(Linker No. 8), -P(O)OR a O-(Linker No. 9), -O-(Linker No. 10), -CR b 2-(Linker number 11), -[(CR b 2) 1-12 O] 1-50 -(Linker No. 12), -C(O)-(Linker No. 13), -C(S)-(Linker No. 14), -C(=N-OH)-(Linker No. 15), -C(NR a )-(Linker No. 16), -C(NH2Cl)-(Linker No. 17), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)S-(Linker No. 20), -SC(O)-(Linker No. 21), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -C(O)NR a SO2- (linker number 24), -SO2NR a C(O)-(Linker No. 25), -OC(O)O-(Linker No. 26), -OC(O)S-(Linker No. 27), -SC(O)O-(Linker No. 28), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (Linker No. 30), -SC(O)NR a -(Linker No. 31), -NR a C(O)S- (Linker No. 32), -S(O) t N(R a )-(wherein t is 1 or 2) (Linker No. 33), -N(R a )S(O) t -(wherein t is 1 or 2) (Linker No. 34), and -X AA - (Linker No. 35).

[0011] In some embodiments, the linker L is an optionally substituted C-C 16 Alkylene (Linker No. 46), -C≡C- (Linker No. 339), -CR a =CR a-(Linker No. 38), optionally substituted phenylene (Linker No. 47), optionally substituted C3-C6 cycloalkylene (Linker No. 48), -[CH2CH2O] 1-16 -(Linker No. 49), -[CH2CH2CH2O] 1-16 -(Linker Number 50), optionally substituted 5- to 6-membered heteroarylene (Linker Number 51), optionally substituted 5- to 20-membered heterocycloalkylene (Linker Number 52), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR a O-(Linker No. 8), -O-(Linker No. 10), -CR b 2-(Linker No. 11), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (linker number 30), and -X AA - (Linker No. 35).

[0012] In some embodiments, the linker L is —[C(R b )2] 1-16 -(Linker No. 341), -C≡C-(Linker No. 339), -CR a =CR a -(Linker No. 38), -[CH2CH2O] 1-16 -(Linker No. 49), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR aO-(Linker No. 7), -O-(Linker No. 10), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (Linker No. 30), -X AA -(linker number 35), TIFF2026501556000002.tif129165; X 1 , X 2 and X 3 independently for each occurrence, NR a , N, C.R. b , S and O.

[0013] In some embodiments, the linker L is of formula (L-1): TIFF2026501556000003.tif18165 formula (L-1), L A is a linking moiety through which A is covalently attached to L'; L' is a bond, or optionally substituted alkylene, optionally substituted alkenylene, optionally substituted alkynylene, optionally substituted arylene, optionally substituted cycloalkylene, optionally substituted heteroalkylene, optionally substituted heteroarylene, optionally substituted heterocycloalkylene, -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -S(O)-(Linker No. 5), -S(O)2-(Linker No. 6), -OP(O)OR a -(Linker number 7), -OP(O)OR a O-(Linker No. 8), -P(O)OR aO-(Linker No. 9), -O-(Linker No. 10), -CR b 2-(Linker number 11), -[(CR b 2) 1-12 O] 1-50 -(Linker No. 12), -C(O)-(Linker No. 13), -C(S)-(Linker No. 14), -C(=N-OH)-(Linker No. 15), -C(NR a )-(Linker No. 16), -C(NH2Cl)-(Linker No. 17), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)S-(Linker No. 20), -SC(O)-(Linker No. 21), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -C(O)NR a SO2- (linker number 24), -SO2NR a C(O)-(Linker No. 25), -OC(O)O-(Linker No. 26), -OC(O)S-(Linker No. 27), -SC(O)O-(Linker No. 28), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (Linker No. 30), -SC(O)NR a -(Linker No. 31), -NR a C(O)S- (Linker No. 32), -S(O) t N(R a )-(wherein t is 1 or 2) (Linker No. 33), -N(R a )S(O) t -(wherein t is 1 or 2) (Linker No. 34), and -X AA -(Linker No. 35); and L P is a linking moiety through which P is covalently attached to L'.

[0014] In some embodiments, the linker L comprises at least one cleavable moiety. In some embodiments, the cleavable moiety comprises an acid-labile moiety, a reducibly-labile moiety, or an enzymatically-labile moiety. In some embodiments, the cleavable moiety comprises one or more groups selected from the following: TIFF2026501556000004.tif155165In formula, Each R a’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

[0015] In some embodiments, the cleavable moiety comprises the reductively labile moiety -SS- (Linker No. 340).

[0016] In some embodiments, the linker L is of formula (L-10): TIFF2026501556000005.tif18165 formula (L-10), L A is the bond, -NR a’ -S-; L 1 is a bond or an optionally substituted C1 to C 18 Alkylene (Linker No. 36), -C≡C- (Linker No. 339), -CR a =CR a (Linker Number 38)-, optionally substituted 6- to 14-membered arylene (Linker Number 39), optionally substituted C3-C 20 Cycloalkylene (linker number 40), -CHO] 1-18 -(Linker No. 41), -[CH2CH2O] 1-18 -(Linker No. 42), -[CH2CH2CH2O] 1-18 -(Linker Number 43), optionally substituted 5- to 18-membered heteroarylene (Linker Number 44), optionally substituted 3- to 20-membered heterocycloalkylene (Linker Number 45), -NR a-(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR a O-(Linker No. 8), -O-(Linker No. 10), -CR b 2-(Linker No. 11), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)S-(Linker No. 20), -SC(O)-(Linker No. 21), -C(O)NR a (Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -OC(O)S-(Linker No. 27), -SC(O)O-(Linker No. 28), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (Linker No. 30), -SC(O)NR a -(Linker No. 31), -NR a C(O)S- (linker number 32), and -X AA - (Linker No. 35); L C is selected from an acid labile moiety, a reductively labile moiety, and an enzymatically labile moiety; L 2 is a bond or an optionally substituted C1 to C 18 Alkylene (Linker No. 36), -C≡C- (Linker No. 339), -CR a =CR a (Linker Number 38)-, optionally substituted 6- to 14-membered arylene (Linker Number 39), optionally substituted C3-C 20 Cycloalkylene (linker number 40), -CHO] 1-18 -(Linker No. 41), -[CH2CH2O] 1-18 -(Linker No. 42), -[CH2CH2CH2O] 1-18-(Linker Number 43), optionally substituted 5- to 18-membered heteroarylene (Linker Number 44), optionally substituted 3- to 20-membered heterocycloalkylene (Linker Number 45), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR a O-(Linker No. 8), -O-(Linker No. 10), -CR b 2-(Linker No. 11), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)S-(Linker No. 20), -SC(O)-(Linker No. 21), -C(O)NR a (Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -OC(O)S-(Linker No. 27), -SC(O)O-(Linker No. 28), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (Linker No. 30), -SC(O)NR a -(Linker No. 31), -NR a C(O)S- (linker number 32), and -X AA - (Linker No. 35); L P is the bond, -NR a’ -(Linker No. 1), -S-(Linker No. 4), and -O-(Linker No. 10); Each R a is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R a’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; Each R b are independently hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -C(O)NR a 2. -CO2R a , -NR a 2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent R b groups taken together form an optionally substituted cycloalkyl, an optionally substituted aryl, an optionally substituted heterocycloalkyl, or an optionally substituted heteroaryl; and -X AA - is an amino acid sequence containing 1 to 4 amino acid moieties.

[0017] In some embodiments, L C teeth Selected from TIFF2026501556000006.tif122165, During the ceremony, Each R a’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl.

[0018] In some embodiments, L C is -SS- (linker number 340).

[0019] In some embodiments, the linker L is of formula (L-11): TIFF2026501556000007.tif27165 formula (L-11), L A is selected from a bond, —NH—, and —S—; L 1 is a bond or -[C(R b )2] 1-16 -(Linker No. 341), -C≡C-(Linker No. 339), -CR a =CR a -(Linker No. 38), -[CH2CH2O] 1-16 -(Linker No. 49), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR a O-(Linker No. 7), -O-(Linker No. 10), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -X AA -(Linker No. 35), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (linker number 30), TIFF2026501556000008.tif88165; L 2 is a bond or -[C(R b )2] 1-16 -(Linker No. 341), -C≡C-(Linker No. 339), -CR a =CR a -(Linker No. 38), -[CH2CH2O] 1-16 -(Linker No. 49), -NR a -(Linker number 1), -N=CR a -(Linker number 2), -CR a =N-(Linker No. 3), -S-(Linker No. 4), -OP(O)OR aO-(Linker No. 7), -O-(Linker No. 10), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -X AA -(Linker No. 35), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (linker number 30), TIFF2026501556000009.tif88165; L P is the bond, -NR a’ -O-; each R1 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R1 groups together form an optionally substituted cycloalkyl; each R2 is independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R2 groups together form an optionally substituted cycloalkyl; Each R ais independently selected at each occurrence from hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, and optionally substituted 5-6 membered heteroaryl; Each R a’ is independently selected at each occurrence from hydrogen and optionally substituted alkyl; Each R b are independently defined for each occurrence as hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2R a , -NR a 2, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl; or two independent R b groups taken together form an optionally substituted cycloalkyl; and -X AA - is an amino acid sequence containing 2 to 4 amino acid moieties.

[0020] In some embodiments, the linker L is of formula (L-12): TIFF2026501556000010.tif37165 formula (L-12), L A is selected from a bond and -NH-; L 1’ is -[C(R b )2] 1-10 -(Linker number 342), -[CH2CH2O] 1-10 -(Linker No. 343), -NR a -(Linker No. 1), -O-(Linker No. 10), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NRa -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -X AA -(Linker No. 35), -OC(O)NR a -(linker number 29), -NR a C(O)O- (linker number 30), TIFF2026501556000011.tif76165; L 2’ is -[C(R b )2] 1-10 -(Linker number 342), -[CH2CH2O] 1-10 -(Linker No. 343), -NR a -(Linker No. 1), -O-(Linker No. 10), -C(O)-(Linker No. 13), -C(O)O-(Linker No. 18), -OC(O)-(Linker No. 19), -C(O)NR a -(Linker No. 22), -NR a C(O)-(Linker No. 23), -OC(O)O-(Linker No. 26), -X AA -(Linker No. 35), -OC(O)NR a -(Linker No. 29), -NR a C(O)O- (linker number 30), TIFF2026501556000012.tif76165; L P is a bond, and -NR a’ (Linker No. 1); each R1 is independently selected from hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl; or both R1 groups together form an optionally substituted C3-C6 cycloalkyl; each R2 is independently selected from hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl; or both R2 groups together form an optionally substituted C3-C6 cycloalkyl; Each R a is independently selected at each occurrence from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 fluoroalkyl, and optionally substituted C3-C6 cycloalkyl; Each R a’ is independently selected at each occurrence from hydrogen and optionally substituted C1-C6 alkyl; Each R b are independently defined for each occurrence as hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2R a , -NR a 2, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 fluoroalkyl, and optionally substituted C3-C6 cycloalkyl; or two independent R b groups taken together form an optionally substituted C3-C6 cycloalkyl; and -X AA - is an amino acid sequence containing two or three amino acid moieties.

[0021] In some embodiments, at least one R1 or R2 is other than hydrogen. In some embodiments, at least one R1 is optionally substituted C1-C8 alkyl. In some embodiments, each R1 is independently optionally substituted C1-C8 alkyl. In some embodiments, at least one R2 is optionally substituted C1-C8 alkyl. In some embodiments, each R2 is independently optionally substituted C1-C8 alkyl.

[0022] In some embodiments, the linker L is Selected from TIFF2026501556000013.tif56165.

[0023] In some embodiments, the linker L is Selected from TIFF2026501556000014.tif177165TIFF2026501556000015.tif158165.

[0024] In some embodiments, the linker is a cleavable linker.

[0025] In some embodiments, the linker L is of formula (L-20): TIFF2026501556000016.tif18165 formula (L-20), L A is the bond, -NR a’ -(Linker No. 1), and -S-(Linker No. 4); L 3 is a bond or -[C(R b )2] 1-8 -, -NR a -, -C(O)-, -C(S)-, -C(NR a )-, -C(NH2Cl)-, -C≡C-, -CR a =CR a -, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C 20 containing one or more groups selected from cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene; L X is optionally substituted C1 to C 18 Alkylene, -C≡C-, -CR a =CR a -, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C 20 Cycloalkylene, -[CHO] 1-18-, -[CH2CH2O] 1-18 -, -[CH2CH2CH2O] 1-18 -, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NR a -, -S-, -O-, -CR b 2-, -C(O)-, -C(S)-, -C(NR a )-, -C(NH2Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR a - and -NR a C(O)—; L 4 is a bond or -[C(R b )2] 1-8 -, -NR a -, -C(O)-, -C(S)-, -C(NR a )-, -C(NH2Cl)-, -C≡C-, -CR a =CR a -, optionally substituted 6- to 14-membered arylene, optionally substituted C3-C 20 containing one or more groups selected from cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene; L P is the bond, -NR a’ -S-, and -O-; Each R a is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R a’is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; and Each R b are independently hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -C(O)NR a 2. -CO2R a , -NR a 2, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent R b The groups taken together form an optionally substituted cycloalkyl, an optionally substituted aryl, an optionally substituted heterocycloalkyl or an optionally substituted heteroaryl.

[0026] In some embodiments, the linker L is of formula (L-21): TIFF2026501556000017.tif24165 formula (L-21), L A is selected from a bond and -NH-; L X is an optionally substituted —[C(R b )2] 1-16 -, -C≡C-, -CR a =CR a -, -[CH2CH2CH2O] 1-16 -, -NR a- , -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR a -, -NR a C(O)-, TIFF2026501556000018.tif57165; L P is a bond and -NR a’ Selected from; Each R a is independently selected at each occurrence from hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, and optionally substituted 5-6 membered heteroaryl; Each R a’ is independently selected at each occurrence from hydrogen and optionally substituted C1-C6 alkyl; and Each R b are independently defined for each occurrence as hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2R a , -NR a 2, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl; or two independent R b The groups taken together form an optionally substituted cycloalkyl.

[0027] In some embodiments, the linker L is of formula (L-22a) or (L-22b): TIFF2026501556000019.tif31165TIFF2026501556000020.tif31165In equations (L-22a) and (L-22b), L A is selected from a bond, and -NH-; L X is an optionally substituted —[C(R b )2] 1-10 -, -C≡C-, -CR a =CR a-, -[CH2CH2CH2O] 1-10 -, -NR a -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR a -, -NR a C(O)-, TIFF2026501556000021.tif57165; L P is a bond, and -NR a’ Selected from; Each R a is independently selected at each occurrence from hydrogen, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, and optionally substituted 5-6 membered heteroaryl; Each R a’ is independently selected at each occurrence from hydrogen and optionally substituted C1-C6 alkyl; and Each R b are independently defined for each occurrence as hydrogen, halides, -OH, -SO3H, -OPO3H2, -PO3H2, -CO2R a , -NR a 2, optionally substituted C1-C8 alkyl, optionally substituted C1-C8 fluoroalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5-10 membered heterocycloalkyl, optionally substituted 5-6 membered heteroaryl; or two independent R b The groups taken together form an optionally substituted cycloalkyl.

[0028] In some embodiments, the linker L is TIFF2026501556000022.tif177165TIFF2026501556000023.tif211165TIFF202650155600 0024.tif210165TIFF2026501556000025.tif198165TIFF2026501556000026.tif186165TI FF2026501556000027.tif215165TIFF2026501556000028.tif203165TIFF20265015560000 29.tif206165TIFF2026501556000030.tif214165TIFF2026501556000031.tif217165TIFF 2026501556000032.tif191165TIFF2026501556000033.tif213165TIFF2026501556000034 .tif198165TIFF2026501556000035.tif187165TIFF2026501556000036.tif191165TIFF20 Selected from 26501556000037.tif184165TIFF2026501556000038.tif213165TIFF2026501556000039.tif218165TIFF2026501556000040.tif192165TIFF2026501556000041.tif152165

[0029] In some embodiments, the linker comprises a non-cleavable linker.

[0030] In some embodiments, -X AA Each amino acid moiety of -X is independently selected from alanine (Ala), arginine (Arg), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), phenylalanine (Phe), tryptophan (Trp), tyrosine (Tyr), valine (Val), citrulline (Cit), and homocitrulline (HoCit). AAwherein each amino acid moiety is independently selected from alanine (Ala), glycine (Gly), lysine (Lys), phenylalanine (Phe), valine (Val), and citrulline (Cit). In some embodiments, the amino acid sequence -X AA - is -Val-Cit- (linker number 319), -Cit-Val- (linker number 320), -Val-Ala- (linker number 321), -Ala-Val- (linker number 322), -Phe-Lys- (linker number 323), -Lys-Phe- (linker number 324), -Ala-Ala- (linker number 325), -Val-Val- (linker number 326), -Gly-Gly- (linker number 327), -Ala-Ala- In some embodiments, the amino acid sequence -X is selected from the group consisting of Ia-Ala- (Linker No. 328), -Gly-Gly-Gly- (Linker No. 329), -Gly-Gly-Phe-Gly- (Linker No. 330), -Gly-Phe-Gly-Gly- (Linker No. 331), -Gly-Gly-Gly-Phe- (Linker No. 332), -Phe-Gly-Gly-Gly- (Linker No. 333), and -Gly-Gly-Gly-Gly- (Linker No. 334). AA - is selected from -Val-Cit- (linker number 319), Cit Val (linker number 320), Val Ala (linker number 321), Ala Val (linker number 322), Phe Lys (linker number 323), Lys Phe (linker number 324), Ala Ala (linker number 325), Val Val (linker number 326), Gly Gly (linker number 327), Ala Ala (linker number 328), and Gly Gly (linker number 329).

[0031] In some embodiments, the payload moiety P is a drug moiety. In some embodiments, the drug moiety is selected from any of the following: TIFF2026501556000042.tif191165TIFF2026501556000043.tif221165TIFF202 6501556000044.tif206165TIFF2026501556000045.tif226165TIFF20265015560 00046.tif189165TIFF2026501556000047.tif226165TIFF2026501556000048.t if175165TIFF2026501556000049.tif184165TIFF2026501556000050.tif132165

[0032] In some embodiments, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 9, the CDR2 comprises the amino acid sequence of SEQ ID NO: 10, the CDR3 comprises the amino acid sequence of SEQ ID NO: 11, and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, the CDR2 comprises the amino acid sequence of SEQ ID NO: 4, and the CDR3 comprises the amino acid sequence of SEQ ID NO: 5.

[0033] In some embodiments, the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 29, the CDR2 comprises the amino acid sequence of SEQ ID NO: 10, and the CDR3 comprises the amino acid sequence of SEQ ID NO: 11, and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, the CDR2 comprises the amino acid sequence of SEQ ID NO: 26, and the CDR3 comprises the amino acid sequence of SEQ ID NO: 5.

[0034] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 21 and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 14. In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a full-length light chain (LC) comprising the amino acid sequence of SEQ ID NO: 20 and a full-length heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 13.

[0035] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a light chain variable domain (VL) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO: 85), 3E10-VL-H2 (SEQ ID NO: 86), 3E10-VL-H3 (SEQ ID NO: 87), 3E10-VL-H4 (SEQ ID NO: 88), 3E10-VL-H5 (SEQ ID NO: 89), and 3E10-VL-H6 (SEQ ID NO: 90). L ), and a heavy chain variable domain (VH) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO: 64), 3E10-VH-H2 (SEQ ID NO: 65), 3E10-VH-H3 (SEQ ID NO: 66), 3E10-VH-H4 (SEQ ID NO: 67), 3E10-VH-H5 (SEQ ID NO: 68), 3E10-VH-H6 (SEQ ID NO: 69), and 3E10-VH-H7 (SEQ ID NO: 70). H ) and

[0036] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO: 85), 3E10-VL-H2 (SEQ ID NO: 86), 3E10-VL-H3 (SEQ ID NO: 87), 3E10-VL-H4 (SEQ ID NO: 88), 3E10-VL-H5 (SEQ ID NO: 89), and 3E10-VL-H6 (SEQ ID NO: 90). L ), and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO: 64), 3E10-VH-H2 (SEQ ID NO: 65), 3E10-VH-H3 (SEQ ID NO: 66), 3E10-VH-H4 (SEQ ID NO: 67), 3E10-VH-H5 (SEQ ID NO: 68), 3E10-VH-H6 (SEQ ID NO: 69), and 3E10-VH-H7 (SEQ ID NO: 70). H ) and

[0037] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof is selected from the group consisting of: (a) VL1 (SEQ ID NO: 85) and VH1 (SEQ ID NO: 64), (b) VL1 (SEQ ID NO: 85) and VH2 (SEQ ID NO: 65), (c) VL1 (SEQ ID NO: 85) and VH3 (SEQ ID NO: 66), (d) VL1 (SEQ ID NO: 85) and VH4 (SEQ ID NO: 67), (e) VL2 (SEQ ID NO: 86) and VH1 (SEQ ID NO: 68), (f) VL2 (SEQ ID NO: 86) and VH1 (SEQ ID NO: 69), (g) VL3 (SEQ ID NO: 87) and VH4 (SEQ ID NO: 69), (h) VL4 (SEQ ID NO: 87) and VH5 (SEQ ID NO: 69), (i) VL5 (SEQ ID NO: 87) and VH6 (SEQ ID NO: 69), (j) VL6 (SEQ ID NO: 87) and VH7 (SEQ ID NO: 69), (j) VL7 (SEQ ID NO: 87) and VH8 (SEQ ID NO: 69), (k) VL8 (SEQ ID NO: 87) and VH9 (SEQ ID NO: 69), (j) VL9 (SEQ ID NO: 87) and VH1 (SEQ ID NO: 69), (j) VL9 (SEQ ID NO: 87) and VH1 (SEQ ID NO: 69), (k ... 64), (f) VL2 (SEQ ID NO: 86) and VH2 (SEQ ID NO: 65), (g) VL2 (SEQ ID NO: 86) and VH3 (SEQ ID NO: 66), (h) VL2 (SEQ ID NO: 86) and VH4 (SEQ ID NO: 67), (i) VL3 (SEQ ID NO: 87) and VH1 (SEQ ID NO: 64), (j) VL3 (SEQ ID NO: 87) and VH2 (SEQ ID NO: 65), (k) VL3 (SEQ ID NO: 87) and VH3 (SEQ ID NO: 66), (l) The VL / VH pair comprises a VL / VH pair selected from the group consisting of: (i) VL3 (SEQ ID NO: 87) and VH4 (SEQ ID NO: 67), (m) VL4 (SEQ ID NO: 88) and VH1 (SEQ ID NO: 64), (n) VL4 (SEQ ID NO: 88) and VH2 (SEQ ID NO: 65), (o) VL4 (SEQ ID NO: 88) and VH3 (SEQ ID NO: 66), (p) VL4 (SEQ ID NO: 88) and VH4 (SEQ ID NO: 67), (q) VL5 (SEQ ID NO: 89) and VH5 (SEQ ID NO: 68), (r) VL5 (SEQ ID NO: 89) and VH6 (SEQ ID NO: 69), (s) VL6 (SEQ ID NO: 90) and VH5 (SEQ ID NO: 68), and (t) VL6 (SEQ ID NO: 90) and VH6 (SEQ ID NO: 69).

[0038] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a light chain variable domain (VL) comprising 3E10-VL-H6 (SEQ ID NO: 90) and a heavy chain variable domain (VH) comprising 3E10-VH-H6 (SEQ ID NO: 69).

[0039] In some embodiments, the antibody, antigen-binding fragment thereof, or cell-permeable fragment thereof comprises a light chain variable domain (VL) comprising the amino acid sequence (DIQMTQSPSSLSASLGDRATITCRASKTVSTSSYSYMHWYQQKPGQPPKLLIKYASYLESGVPSRFSGSGSGTDFTLTISSLQPEDAATYYCQHSREFPWTFGGGTKVEIK) (SEQ ID NO: 126) and a heavy chain variable domain (VH) comprising the amino acid sequence (EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVSYISSGSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGLLLDYWGQGTTVTVSS) (SEQ ID NO: 127).

[0040] In embodiments, the present disclosure provides a method of treating a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a conjugate of the present disclosure. [Brief explanation of the drawings]

[0041] [Figure 1A] 1 shows a conjugation strategy for preparing an antibody-drug conjugate (ADC) according to one embodiment of the present disclosure. It is a schematic diagram showing labeling of a 3E10 antibody, such as the chimeric 3E10-cD31N antibody (cD31N), with a 4-(N-maleimidomethyl)cyclohexane-1-carboxylate succinimidyl (SMCC) linker (Step 1, SMCC labeling), followed by conjugation of a maytansinoid DM1 linker to a warhead (N2′-deacetyl-N2′-(3-mercapto-1-oxopropyl)-maytansine) (DM1) (Step 2, DM1 labeling). [Figure 1B] 1 shows a conjugation strategy for preparing an antibody-drug conjugate (ADC) according to one embodiment of the present disclosure, and provides further details regarding ADC synthesis. [Figure 2] 10 is a high performance liquid chromatography (HPLC) chromatogram measuring the amount and size of DM1, cD31N (3E10-cD31N antibody), and the cD31N-DM1 complex after 14 days of incubation at 4° C. [Figure 3] Shown is the viability of B16 melanoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 3 days. [Figure 4] 1 shows the viability of Capan-1 pancreatic cancer cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 6 days. [Figure 5] Shown is the viability of DAYO medulloblastoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 3 days. [Figure 6] Shown is the viability of GL261 glioma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 6 days. [Figure 7] (A) Viability of wild-type BRCA2 (WT or BRCA2wt / wt) DLD-1 colon adenocarcinoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 6 days. (B) Viability of BRCA2-deficient (BRCA2- / -) DLD-1 colon adenocarcinoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 6 days. [Figure 8] (A) Viability of PEO-4 (WT) ovarian adenocarcinoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 5 days. (B) Viability of PEO-1 (BRCA2- / -) ovarian adenocarcinoma cells treated with various concentrations of DM1 alone or the cD31N-DM1 complex after 5 days. [Figure 9] 1 shows a conjugation strategy for preparing an antibody-drug conjugate (ADC) according to one embodiment of the present disclosure, showing conjugation of the 3E10 antibody with an O-succinyl linker and the anti-tumor drug SN38. [Figure 10] Western blots showing the results of treating WT and BRCA2- / - DLD-1 cells with various concentrations of SN38 alone, wild-type 3E10 chimeric antibody (cWT), cWT-SN38 complex, cD31N antibody, or cD31N-SN38 complex. The protein gamma (γ)-H2AX is a marker of DNA damage. [Figure 11](A) Viability of WT DLD-1 cells treated with various concentrations of SN38 alone, cWT-SN38 complex, or cD31N-SN38 complex. (B) Viability of BRCA2- / - DLD-1 cells (Figure 11B) treated with various concentrations of SN38 alone, cWT-SN38 complex, or cD31N-SN38 complex. [Figure 12] Figure 1 shows the results of treatment of an in vivo mouse model of melanoma with the cD31N-SN38 conjugate. (A) Tumor growth over time is shown after treatment of mice with 12.5 mg / kg of the cD31N-SN38 conjugate (n=4) or phosphate-buffered saline (PBS) control (n=10). (B) Post-implant weight over time is shown for mice treated with the cD31N-SN38 conjugate compared to mice treated with PBS. [Figure 13] 1 shows the timeline of the treatment duration experiment described herein. [Figure 14] Cell viability of WT and BRCA2- / - DLD-1 cells treated with SN38, cWT-SN38, or cD31N-SN38 is shown on days 3, 5, and 7, respectively. (A) Viability of WT DLD-1 cells is shown. (B) Viability of BRCA2- / - DLD-1 cells is shown. Calculated EC50 values ​​are provided. [Figure 15] Cell viability of WT and BRCA2- / - DLD-1 cells treated with SN38, cWT-SN38, or cD31N-SN38 is shown on days 3, 5, and 7, respectively. (A) Viability of WT DLD-1 cells is shown. (B) Viability of BRCA2- / - DLD-1 cells is shown. Calculated EC50 values ​​are provided. [Figure 16] Cell viability of WT and BRCA2- / - DLD-1 cells treated with SN38, cWT-SN38, or cD31N-SN38 is shown on days 3, 5, and 7, respectively. (A) Viability of WT DLD-1 cells is shown. (B) Viability of BRCA2- / - DLD-1 cells is shown. Calculated EC50 values ​​are provided. [Figure 17]The effects of short-term and long-term treatment on cell viability in WT DLD-1 cells (Figures 17A and 17B) on day 7 after treatment with SN38 or cD31N-SN38 complexes are shown. Short-term treatment was defined as 24 hours of treatment followed by a change to fresh, untreated medium. Long-term treatment was defined as 7 days of treatment without a change of medium. Calculated EC50 values ​​are provided. [Figure 18] The effects of short-term and long-term treatment on cell viability in BRCA2- / - DLD-1 cells (Figures 18A and 18B) after 7 days of treatment with SN38 or cD31N-SN38 complex are shown. Short-term treatment was defined as 24 hours of treatment followed by a change to fresh, untreated medium. Long-term treatment was defined as 7 days of treatment without changing the medium. Calculated EC50 values ​​are provided. [Figure 19] Figure 19 shows the results of clonogenic survival assays in WT (Figure 19A) and BRCA2- / - (Figure 19B) DLD-1 cells after treatment with various concentrations of SN38, cWT antibody, or cWT-SN38 complex. Data are presented as % survival normalized to no treatment (NT). [Figure 20] (A) Shown are the results of a clonogenic survival assay in WT (FIG. 20A) DLD-1 cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38 complex. Data are shown as % survival normalized to no treatment (NT). (B) Shown are the results of a clonogenic survival assay in BRCA2- / - (FIG. 20B) DLD-1 cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38 complex. Data are shown as % survival normalized to no treatment (NT). [Figure 21A] Western blot of specific DNA damage response proteins in WT and BRCA2- / - DLD-1 cells after treatment with SN38, cWT antibody, cD31N antibody, cWT-SN38 complex, or cD31N-SN38 complex. Results are shown for DLD-1 cells harvested 24 hours after treatment. [Figure 21B]Western blot of specific DNA damage response proteins in WT and BRCA2- / - DLD-1 cells after treatment with SN38, cWT antibody, cD31N antibody, cWT-SN38 complex, or cD31N-SN38 complex. Results are shown for DLD-1 cells harvested 48 hours after treatment. [Figure 22] Western blot of specific DNA damage response proteins in WT and BRCA2- / - DLD-1 cells after treatment with SN38, cWT antibody, cD31N antibody, cWT-SN38 complex, or cD31N-SN38 complex. Results are shown for DLD-1 cells harvested 96 hours after treatment (Figure 22). [Figure 23A] Figure 1 shows the cell viability of wild-type and IDH1 mutant HCT116 colon cancer cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38. Figure 2 shows the viability of wild-type HCT116 cells after 5 days of treatment. [Figure 23B] Figure 1 shows the cell viability of WT and IDH1 mutant HCT116 colon cancer cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38. Figure 2 shows the viability of IDH1 mutant HCT116 cells on day 5 of treatment. [Figure 24A] Figure 1 shows cell viability of wild-type and IDH1 mutant HCT116 colon cancer cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38. Figure 2 shows viability of wild-type HCT116 cells after 7 days of treatment. Calculated EC50 values ​​are provided. [Figure 24B] Figure 1 shows the cell viability of WT and IDH1 mutant HCT116 colon cancer cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38. Figure 2 shows the viability of IDH1 mutant HCT116 cells on day 7 of treatment. Calculated EC50 values ​​are provided. [Figure 25A] Figure 1 shows the results of a clonogenic survival assay in WT HCT116 cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38 conjugate. Data are presented as % survival normalized to no treatment (NT). [Figure 25B]

[0023] Figure 1 shows the results of a clonogenic survival assay in IDH1 mutant HCT116 cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38 conjugate. Data are presented as % survival normalized to no treatment (NT). [Figure 26] Western blot of DNA damage response proteins in WT and IDH1 mutant HCT116 cells after treatment with 100 nM SN38, humanized 3E10 antibody V66, or V66-SN38 complex. [Figure 27] 1 shows the viability of LoVo colon cancer cells treated with SN38, cD31N antibody, cD31N Fab antibody fragment, cD31N-SN38 complex, cDN31-Fab-SN38 complex, V66 antibody, V66 Fab antibody fragment, V66-SN38 complex, and V66-Fab-SN38 complex. [Figure 28] 1 shows the viability of SKOV3 ovarian cancer cells after treatment with various concentrations of SN38, cD31N antibody, or cD31N-SN38 conjugate for 7 days. Calculated EC50 values ​​are provided. [Figure 29] 1 shows the results of treatment of SKOV3 ovarian cancer cells with SN38, cD31N antibody, cD31N Fab antibody fragment, cD31N-SN38 conjugate, cDN31-Fab-SN38 conjugate, V66 antibody, V66 Fab antibody fragment, V66-SN38 conjugate, and V66-Fab-SN38 conjugate. [Figure 30] Western blot of DNA damage response proteins in SKOV3 ovarian cancer cells after treatment with 100 nM SN38, cD31N-SN38 complex, and 3E10-cD31N antibody for 24, 48, and 72 hours. [Figure 31A] Figure 1 shows the cell viability of PEO-4 and PEO-1 ovarian cancer cells treated chronically with various concentrations of SN38, cD31N antibody, or cD31N conjugate at day 7. Figure 2 shows the cell viability of PEO-1 cells. Calculated EC50 values ​​are provided. [Figure 31B]Figure 1 shows the cell viability of PEO-4 and PEO-1 ovarian cancer cells treated chronically with various concentrations of SN38, cD31N antibody, or cD31N conjugate at day 7. Figure 2 shows the cell viability of PEO-4 cells. Calculated EC50 values ​​are provided. [Figure 32A] 1 shows Western blots of DNA damage response proteins in PEO-4 and PEO-1 cells after treatment with 100 nM SN38, cD31N antibody, or cD31N-SN38 complex. Protein levels are shown 24 hours after treatment. [Figure 32B] 1 shows Western blots of DNA damage response proteins in PEO-4 and PEO-1 cells after treatment with 100 nM SN38, cD31N antibody, or cD31N-SN38 complex, with protein levels shown 48 hours after treatment. [Figure 33A] Figure 1 shows the results of an in vivo colon cancer model mouse treated with systemic administration of cD31N-SN38 conjugate for four consecutive days followed by two days of no treatment, or two consecutive days of systemic administration followed by five days of no treatment. Tumor growth over time is shown after mice were treated with 12.5 mg / kg of cD31N-SN38 conjugate or phosphate-buffered saline (PBS) control. [Figure 33B] Figure 1 shows the results of an in vivo colon cancer model mouse treated with systemic administration of cD31N-SN38 conjugate for four consecutive days followed by two days of no treatment, or two consecutive days of systemic administration followed by five days of no treatment. The weight of mice treated with cD31N-SN38 conjugate over time after implantation is shown compared to mice treated with PBS. [Figure 34] 1 shows the in vitro potency of 3E10-SN38 ADC against various cancer cell lines at nanomolar concentrations. [Figure 35] Figure 1 shows the in vitro potency of 3E10-DM1 ADC against various cancer cell lines at picomolar concentrations. [Figure 36A] The efficacy of cD31N-drug conjugates (10 mg / kg) in vivo is demonstrated using a mouse cancer model. [Figure 36B]The efficacy of cD31N-drug conjugates (10 mg / kg) in vivo is demonstrated using a mouse cancer model. [Figure 36C] The efficacy of cD31N-drug conjugates (10 mg / kg) in vivo is demonstrated using a mouse cancer model. [Figure 37] 1 shows the viability of human Dukes-C colon adenocarcinoma (DLD1) cell lines treated with V66-PNU, V66-MMAE, and V66-SN38 conjugates. [Figure 38A] 1 shows the results of the in vitro efficacy of 3E10 ADC against the ENT2(+) tumor cell line A427. [Figure 38B] 1 shows the results of the in vitro efficacy of 3E10 ADC against the ENT2(+) tumor cell line LOVO. [Figure 38C] 1 shows the results of in vitro efficacy of 3E10 ADC against the ENT2(+) tumor cell line MCF7. [Figure 39A] 1 shows intermediates of Lys-azide conjugation of 3E10-D31N monoclonal antibody (V66) after proteolysis. [Figure 39B] 1 shows mass spectrometry results of mapped intermediates of Lys-azide conjugation of 3E10-D31N monoclonal antibody (V66) after proteolysis. [Figure 39C] The sequences of humanized 3E10-D31N monoclonal antibody (V66) and 3E10-D31N are shown. [Figure 40A] 1 summarizes the improved cellular internalization of 3E10-D31N monoclonal antibody (V66) oligonucleotide conjugates in A427 cells using transglutaminase-mediated enzymatic conjugation. [Figure 40B] 1 summarizes the improved cellular internalization of 3E10-D31N monoclonal antibody (V66) oligonucleotide conjugates in A427 cells using transglutaminase-mediated enzymatic conjugation. [Figure 41A] Tumor volumes in mice treated with V66-MMAE conjugate on a QWx4, 2QWx4, or 3QWx4 schedule are shown. [Figure 41B]Shown are the body weights of mice treated with V66-MMAE conjugates on a QWx4, 2QWx4, or 3QWx4 schedule. [Figure 42A] Tumor volumes in PBS-treated mice are shown. [Figure 42B] Tumor volumes in mice treated with V66-MMAE conjugate on a QWx4 schedule are shown. [Figure 42C] Tumor volumes in mice treated with V66-MMAE conjugate on a 2QWx4 schedule are shown. [Figure 42D] Tumor volumes in mice treated with V66-MMAE conjugate on a 3QWx4 schedule are shown. [Figure 43A] 1 shows the in vitro efficacy results of the 3E10 ADC against three ENT2(+) tumor cell lines (MCF7, LOVO, and A427). [Figure 43B] 1 shows the in vitro efficacy results of the 3E10 ADC against three ENT2(+) tumor cell lines (MCF7, LOVO, and A427). [Figure 44] The structure of V66-vc-Seco-DUBA ADC is shown. DETAILED DESCRIPTION OF THE INVENTION

[0042] Detailed Description I. Introduction Advantageously, methods and compositions have been developed to target therapeutic agents, e.g., drugs such as antitumor drugs, or oligonucleotides, to various cancer tissues in vivo and facilitate delivery of these therapeutic polynucleotides to diseased cells, e.g., cancer cells that exhibit high levels of ENT2 on their cell surface. Accordingly, the present disclosure provides compositions, conjugates, and methods for delivering therapeutic agents to cancer tissues. In some embodiments, the methods and compositions are specifically used for treating cancer. For example, compositions comprising a conjugate of (i) a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, and (ii) a therapeutic agent, as well as methods of using such compositions for treating cancer, are described.

[0043] The studies described herein demonstrate that ADCs containing the cell-penetrating and nucleic acid-binding 3E10 antibody conjugated to antitumor drugs or oligonucleotides via a linker can effectively penetrate and kill tumor cells with specificity. For example, Example 3 demonstrates the broad applicability and efficacy of the cD31N-DM1 conjugate against various cancer cell lines. The cD31N-DM1 ADC was effective in all cancer cell lines tested, regardless of genetic background or tissue origin, and the ADC-drug was more potent than the free drug during culture in each cell line. This suggests that the targeting of the 3E10 ADC is not limited to a specific cancer antigen or tumor epitope. Advantageously, the 3E10 ADC provided herein can broadly target any cancer based on the cell surface expression of ENT2, a nucleoside transporter overexpressed in many cancers. Example 5 demonstrates that the 3E10-SN38 conjugate can effectively kill colon cancer cells by inducing DNA damage through the delivery of SN38. Example 6 demonstrates that administration of the cD31N-SN38 conjugate suppresses tumor growth in mice without nonspecific toxicity. Example 7 demonstrates that a single administration of SN38 or the 3E10-SN38 conjugate can have long-term effects on cancer cell growth, and these effects are dose-dependent. Example 20 demonstrates that systemic administration of the cD31N-SN38 conjugate after transplantation suppresses tumor growth in mice without nonspecific toxicity. Examples 21 and 22 demonstrate the broad applicability and efficacy of the cD31N-SN38 conjugate and the cD31N-DM1 conjugate, respectively, against various cancer cell lines. Example 24 shows that conjugates of 3E10-D31N monoclonal antibody (V66)-PNU, V66-MMAE, and V66-SN38 can each effectively kill the DLD1 (BRCA2- / -) cancer cell line 7 days after treatment. Example 27 shows that cellular internalization of V66 oligonucleotide conjugates can be improved using transglutaminase-mediated enzymatic conjugation.

[0044] II. Definition The terminology used in this disclosure is for the purpose of describing particular aspects only and is not intended to be limiting.

[0045] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms unless the context clearly indicates otherwise. It will also be understood that the term "and / or," as used herein, refers to and encompasses any and all possible combinations of one or more of the associated listed items. It will be further understood that, unless the context requires otherwise, when used herein, the terms "includes," "comprising," or variations thereof specify the presence of the stated features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. Furthermore, to the extent that the terms "including," "includes," "having," "has," "with," or variations thereof are used in either the detailed description and / or claims, such terms are intended to be inclusive in a manner similar to "comprising." Furthermore, when the terms "comprising," "including," "includes," "having," "has," "with," or variations thereof are used in either the detailed description and / or claims, alternatives described as "consisting of" or "consisting essentially of" are intended to be encompassed in such disclosure.

[0046] The recitation of ranges of values ​​herein is intended to serve merely as a shorthand method of referring individually to each separate value falling within the range, unless otherwise stated herein, and each separate value is incorporated herein as if it were individually set forth herein.

[0047] Use of the term "about" is intended to indicate a value above or below the stated value within a range of about ±10%.

[0048] As used herein, the term "antibody" refers to an immunoglobulin molecule that recognizes and specifically binds to a target, e.g., a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or a combination of the foregoing, through at least one antigen recognition site within the variable region of the immunoglobulin molecule. As used herein, the term "antibody" is used in the broadest sense and encompasses monoclonal antibodies (including full-length monoclonal antibodies), polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), chimeric antibodies, humanized antibodies, human antibodies, fusion proteins containing an antigenic portion of an antibody, and antibody fragments (e.g., Fab, Fab', F(ab')2, Fv fragments, scFv molecules), and any other modified immunoglobulin molecules that contain an antigen recognition site, so long as they exhibit one or more desired biological activities. In embodiments, the "desired biological activity" of an antibody refers to the ability of the antibody to bind to its target antigen, e.g., a nucleic acid, e.g., DNA. In embodiments, the "desired biological activity" may further include the antibody binding to the target antigen and producing a measurable biological response that can be measured in vitro or in vivo. Such activity can be antagonistic or agonistic. In embodiments, the "desired biological activity" of an antibody refers to the ability of the antibody to bind to a target, e.g., a nucleic acid molecule. In embodiments, the "desired biological activity" of an antibody refers to the ability of the antibody to bind to a cellular receptor, e.g., ENT2. In embodiments, the "desired biological activity" of an antibody refers to the ability of the antibody to be internalized by a target cell. As used herein, a "target antigen" refers to a molecule that is specifically bound by the antigen-binding domain comprising the variable region of a given antibody. The term "specifically binds" refers to an antibody that binds to its cognate antigen (e.g., a nucleic acid, e.g., DNA) but does not significantly bind to other antigens.

[0049] Depending on the amino acid sequence of the constant domain of their heavy chains, antibodies (immunoglobulins) can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, some of which can be further divided into subclasses or isotypes, e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. As used herein, "isotype" refers to any of the immunoglobulin subclasses defined by the chemical and antigenic characteristics of the constant region. The heavy chain constant domains corresponding to the different classes of immunoglobulins are called α, γ, ε, δ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known and are generally described, for example, in Abbas et al. Cellular and Mol. Immunology, 4th ed. (WB Saunders, Co., 2000). It should be understood that the antibodies disclosed herein can also include hybrids of isotypes and / or subclasses.

[0050] The antibodies of the present disclosure are generally isolated or recombinant. When used to describe the various polypeptides disclosed herein, "isolated" refers to a polypeptide that has been identified and separated and / or recovered from the cell or cell culture in which it was expressed. Typically, an isolated polypeptide is prepared by at least one purification step. An "isolated antibody" refers to an antibody that is substantially free of other antibodies having different antigen specificities. As used herein, "recombinant antibody" refers to an antibody produced using recombinant nucleic acid techniques in an exogenous host cell; recombinant antibodies may also be isolated.

[0051] "Native antibodies" are typically heterotetrameric glycoproteins of approximately 150,000 daltons composed of two identical light (L) chains and two identical heavy (H) chains. Each light chain is linked to a heavy chain by one covalent disulfide bond, while the number of disulfide bonds varies among the heavy chains of different immunoglobulin isotypes. Each heavy and light chain also has regularly spaced intrachain disulfide bridges. Each heavy chain has a variable domain (VH) at one end followed by several constant domains. Each light chain has a variable domain (VL) at one end and a constant domain at its other end, with the light-chain constant domain aligned with the first constant domain of the heavy chain and the light-chain variable domain aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light-chain variable domain and the heavy-chain variable domain.

[0052] The term "constant domain" refers to the portion of an immunoglobulin molecule that has a more conserved amino acid sequence than the other portion of the immunoglobulin, the variable domain, which contains the antigen-binding site. The constant domain contains the CH1, CH2, and CH3 domains (collectively CH) of the heavy chain and the CHL (or CL) domain of the light chain.

[0053] The "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of the heavy or light chain of the antibody. The heavy chain variable domain may be referred to as "VH." The light chain variable domain may be referred to as "VL." These domains are generally the most variable parts of an antibody and contain the antigen-binding site. The term "variable" refers to the fact that certain portions of the variable domains differ significantly in sequence among antibodies and are used to determine the binding and specificity of each particular antibody for its particular antigen. However, variability is not evenly distributed throughout the variable domains of an antibody. Antigen specificity is concentrated in three segments, called hypervariable regions (HVRs) or complementarity-determining regions (CDRs), in both the light and heavy chain variable domains. The "variable heavy domain" pairs with the "variable light domain" to form the antigen-binding domain (ABD), which specifically binds to the target antigen. The more highly conserved portions of the variable domains are called framework regions (FRs). The variable domains of naturally occurring heavy and light chains each contain four FR regions, which largely adopt a β-sheet structure and are connected by three CDRs / HVRs, which form loops that connect the β-sheet structure and, in some cases, form part of the β-sheet structure. The CDRs / HVRs of each chain are held in close proximity by the FR regions and, together with the CDRs / HVRs of the other chain, contribute to the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, Fifth Edition, National Institutes of Health, Bethesda, Md. (1991)). The constant domains are not directly involved in binding the antibody to an antigen but exhibit various effector functions, such as the participation of the antibody in antibody-dependent cellular cytotoxicity.

[0054] The terms "hypervariable region," "HVR," "HV," "complementarity-determining region," and "CDR," used interchangeably herein, refer to the regions of an antibody variable domain that are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies contain six HVRs or CDRs: three in the VH (H1, H2, H3; or VH CDR1, VH CDR2, VH CDR3) and three in the VL (L1, L2, L3; or VL CDR1, VL CDR2, VL CDR3).

[0055] The "light chains" of antibodies (immunoglobulins) from any mammalian species can be assigned to one of two clearly distinct types, called kappa ("κ") and lambda ("λ"), based on the amino acid sequences of their constant domains.

[0056] The CDRs of the VH and VL domains combine to form the Fv region. In embodiments, the VH and VL domains comprise six CDRs, ABD. In the "Fab" format, the variable heavy chain domain (VH; comprising VH CDR1, VH CDR2, and VH CDR3) and the variable light chain domain (VL or VL; comprising VL CDR1, VL CDR2, and VL CDR3) comprise six sets of CDRs, with the C-terminus of the VH domain linked to the N-terminus of the CH1 domain of the heavy chain and the C-terminus of the VL domain linked to the N-terminus of the constant light chain domain (thus forming the light chain). In the "scFv" format, the VH and VL domains are covalently linked into a single polypeptide sequence, generally via the use of a linker (e.g., an scFv linker), and can have an N- to C-terminal configuration of VH-linker-VL or VL-linker-VH. Generally, the C-terminus of the scFv domain is linked to the N-terminus of the hinge of the second monomer.

[0057] "Fab" or "Fab region," as used herein, generally refers to a polypeptide comprising the VH, CH1, VL, and CL immunoglobulin domains on two different polypeptide chains (e.g., VH-CH1 on one chain and VL-CL on the other chain). Fab can refer to this region alone or can refer to this region in the context of an antibody of the present disclosure. In embodiments, Fab comprises an Fv region in addition to the CH1 CL domain.

[0058] Another part of the heavy chain is the hinge region. As used herein, "hinge," "hinge region," "antibody hinge region," or "hinge domain" refers to a flexible polypeptide comprising the amino acids between the first and second constant domains of an antibody. Structurally, the IgG CH1 domain ends at EU215, and the IgG CH2 domain begins at EU231. Thus, for IgG, the antibody hinge is defined herein as comprising positions 216 (E216 in IgG1) to 230 (p230 in IgG1), with numbering according to the EU index as in Kabat. In some cases, "hinge fragments" containing fewer amino acids at either or both the N- and C-termini of the hinge domain are used.

[0059] As used herein, "heavy chain constant region" refers to the CH1-hinge-CH2-CH3 portion of an antibody or fragment thereof, excluding the variable heavy domain. In embodiments, the heavy chain constant region comprises amino acids 118-447 of human IgG1 according to EU numbering. As used herein, "heavy chain constant region fragment" refers to a heavy chain constant region that contains a few amino acids from either or both the N-terminus and C-terminus, but retains the ability to form a dimer with another heavy chain constant region.

[0060] As used herein, "Fv," "Fv fragment," or "Fv region" refers to a polypeptide comprising the VL and VH domains of an antibody binding domain. The Fv region can take both Fab and scFv formats, in which the VL and VH domains are joined (e.g., by a linker as discussed herein) to form an scFv.

[0061] As used herein, "Fc," "Fc region," or "Fc domain" refers to a polypeptide comprising the CH2-CH3 domain of an IgG molecule, optionally including the hinge. In EU numbering for human IgG1, the CH2-CH3 domain comprises amino acids 231-447, and the hinge is 216-230. Thus, the definition of "Fc domain" includes both amino acids 231-447 (CH2-CH3) and 216-447 (hinge-CH2-CH3) of IgG1, or fragments thereof. An "Fc fragment" in this context can contain fewer amino acids from either or both the N-terminus and C-terminus, yet still retain the ability to dimerize with another Fc domain or Fc fragment, as can be detected using standard methods, generally based on size (e.g., non-denaturing chromatography, size-exclusion chromatography, etc.). In embodiments, the disclosed ADCs comprise a human Fc domain. In embodiments, the disclosed ADCs comprise an Fc domain derived from human IgG1, IgG2, or IgG4.

[0062] A "variant Fc domain" comprises amino acid modifications relative to the parent Fc domain. Thus, a "variant human IgG1 Fc domain" contains amino acid modifications (typically amino acid substitutions, but in the case of ablation mutants, including amino acid deletions) relative to the human IgG1 Fc domain. In embodiments, the variant Fc domain has at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 97%, at least about 98%, or at least about 99% identity to the corresponding parent human IgG1 Fc domain. In embodiments, the percent identity is calculated using the identity algorithm discussed below. In embodiments, the percent identity is calculated using the BLAST algorithm known in the art using default parameters. In embodiments, the variant Fc domain has 1 to about 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) amino acid modifications relative to the parent Fc domain. In embodiments, the variant Fc domain retains the ability to form dimers with the Ir Fc domain, as measured using known techniques described herein, such as non-denaturing gel electrophoresis.

[0063] For all positions discussed in this disclosure related to antibodies, unless otherwise noted, the numbering of amino acid positions follows the EU index. The EU index or EU numbering scheme as in EU index or Kabat refers to EU antibody numbering. Kabat et al. collected a large number of primary sequences of heavy and light chain variable regions. They classified each primary sequence into CDRs and frameworks based on the degree of sequence conservation and compiled a list of them. See SEQUENCES OF IMMUNOLOGICAL INTEREST, 5th edition, NIH Publication, No. 91-3242, EA Kabat et al., Edelman et al., 1969, Proc. Natl. Acad. Sci. USA, 63:78-85, the contents of which are incorporated herein by reference. In embodiments of the present disclosure, the numbering of amino acid positions follows the IMGT system.

[0064] The terms "full-length antibody," "intact antibody," and "whole antibody" are used interchangeably herein to refer to an antibody in its substantially intact form, rather than an antibody fragment, as defined below. These terms specifically refer to antibodies having heavy chains containing an Fc region.

[0065] "Antibody fragments" comprise a portion of an intact antibody, preferably the antigen-binding region thereof. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0066] A "naked antibody" for purposes herein is an antibody that is not conjugated to a cytotoxic moiety or radiolabel.

[0067] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible minor mutations, e.g., naturally occurring mutations. Thus, the modifier "monoclonal" indicates the character of the antibody as not being a mixture of distinct antibodies. In certain embodiments, such monoclonal antibodies typically comprise an antibody comprising a target-binding polypeptide sequence, wherein the target-binding polypeptide sequence was obtained by a process that includes selection of a single target-binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection process can be the selection of a unique clone from a plurality of clones, such as a pool of hybridoma clones, phage clones, or recombinant DNA clones. The selected target-binding sequence can be further modified to, for example, improve affinity for the target, humanize the target-binding sequence, improve its production in cell culture, reduce its in vivo immunogenicity, or create a multispecific antibody, and it should be understood that antibodies comprising modified target-binding sequences are also monoclonal antibodies of the present disclosure. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically uncontaminated by other immunoglobulins.

[0068] The monoclonal antibodies herein specifically include "chimeric" antibodies in which a portion of the heavy and / or light chain is identical to or homologous to corresponding sequences in antibodies from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is identical to or homologous to corresponding sequences in antibodies from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit one or more desired biological activities (see, e.g., U.S. Pat. No. 4,816,567 and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). Typically, the variable regions of both the light and heavy chains correspond to the variable regions of antibodies from one mammal (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and / or capacity, while the constant regions are homologous to sequences of antibodies from another mammal (e.g., human) to avoid eliciting an immune response from that species. Chimeric antibodies include the PRIMATTZED® antibody, the antigen-binding region of which is derived from an antibody produced, for example, by immunizing macaque monkeys with an antigen of interest.

[0069] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In embodiments, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from the recipient's CDR / HVR are replaced by residues from a CDR / HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and / or capacity. In some cases, FR residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies can comprise residues that are not found in the recipient antibody or the donor antibody. These modifications can be made to further refine antibody performance. Generally, humanized antibodies will comprise substantially all of at least one, and usually two, variable domains, with all or substantially all of the hypervariable loops corresponding to those of a non-human immunoglobulin and all or substantially all of the FRs being those of a human immunoglobulin sequence. The humanized antibody also optionally will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. See, e.g., Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions, 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409. Examples of methods used to generate humanized antibodies are described in US Pat. Nos. 5,225,539 or 5,639,641, which are incorporated herein by reference in their entirety.

[0070] As used herein, the term "human antibody" refers to an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and / or produced using any technique known in the art. This definition of a human antibody includes intact or full-length antibodies, fragments thereof, and / or antibodies comprising at least one human heavy and / or light chain polypeptide. This definition of a human antibody specifically excludes humanized antibodies comprising non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also available for preparing human monoclonal antibodies are methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5:368-74 (2001). Human antibodies can be prepared by administering antigen to transgenic animals, e.g., immunized xenomouse, that have been engineered to produce such antibodies in response to antigen challenge but in which the endogenous gene locus has been disabled. (See, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology.) See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies produced by human B cell hybridoma technology.

[0071] A "species-dependent antibody" is an antibody that has a stronger binding affinity for an antigen from a first mammalian species than for a homologue of that antigen from a second mammalian species. Typically, species-dependent antibodies "bind specifically" to a human antigen (e.g., about 1 x 10 -7M or less, preferably about 1 × 10 -8 M or less, more preferably about 1 × 10 -9 The species-dependent antibody has a binding affinity (Kd) value of M or less, or a binding affinity for a homologue of the antigen from a second non-human mammalian species that is at least about 50-fold, or at least about 500-fold, or at least about 1000-fold weaker than the binding affinity for the human antigen. The species-dependent antibody can be any of the various types of antibodies defined above, but is preferably a humanized or human antibody.

[0072] The term "linear antibody" refers to the antibodies described in Zapata et al. (1995), Protein Eng, 8(10):1057-1062. Briefly, these antibodies comprise a pair of tandem Fd segments (VH-CH1-VH-CH1), which, together with complementary light chain polypeptides, form a pair of antigen-binding regions. Linear antibodies can be bispecific or monospecific.

[0073] As used herein, "modification" refers to an amino acid substitution, insertion, deletion, and / or any other mutation in a polypeptide sequence.

[0074] As used herein, a "variant protein" or "protein variant" or "variant" refers to a protein that differs from that of a parent protein by virtue of at least one amino acid modification. A protein variant has at least one amino acid modification compared to the parent protein, but not so many that the variant protein does not align with the parent protein using an alignment program such as those described below. Generally, a variant protein (such as a variant Fc domain, as described herein) is generally at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% identical to the parent polypeptide using alignment programs known in the art, such as BLAST.

[0075] Sequence identity between two similar sequences (e.g., antibody variable domains) can be determined using the "BLAST" algorithm, as described by Smith, T.F. & Waterman, M.S. (1981), "Comparison Of Biosequences," Adv. Appl. Math. 2:482 (local homology algorithm); Needleman, S.B. & Wunsch, C.D. (1970), "A General Method Applicable To The Search For Similarities In The Amino Acid Sequence Of Two Proteins," J. Mol. Biol. 48:443 (homology alignment algorithm); Pearson, W.R. & Lipman, D.J. (1988), "Improved Tools For Biological Sequence Comparison," Proc. Natl. Acad. Sci. (USA) 85:2444 (search by similarity method); or Altschul, S.F. et al. (1990), "Basic Local Alignment Search Tool," J. Mol. Biol. 215:403-10 (BLAST). Algorithms such as the BLAST algorithm are described in detail in the webpage at URL blast.ncbi.nlm.nih.gov / Blast.cgi. When using any of the aforementioned algorithms, default parameters (for window length, gap penalties, etc.) are used. Unless otherwise specified, sequence identity is determined using the BLAST algorithm using default parameters.

[0076] In embodiments, the parent polypeptide, e.g., an Fc parent polypeptide, is a heavy chain constant domain or Fc region from a human wild-type sequence, e.g., IgG1, IgG2, IgG3, or IgG4, although human sequences with variants can also serve as a "parent polypeptide." In embodiments, the antibody sequences described herein have at least about 75%, at least about 76%, at least about 77%, at least about 78%, at least about 79%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or at least about 99.5% sequence identity to the parent polypeptide sequence. Thus, as used herein, "antibody variant" or "variant antibody" refers to an antibody that differs from a parent antibody by at least one amino acid modification; as used herein, "IgG variant" or "variant IgG" refers to an IgG that differs from a parent IgG (e.g., a human IgG sequence) by at least one amino acid modification; as used herein, "immunoglobulin variant" or "variant immunoglobulin" refers to an immunoglobulin sequence that differs from a parent immunoglobulin sequence by at least one amino acid modification; and as used herein, "Fc variant" or "variant Fc" refers to an Fc that differs from the Fc domain of a parent Fc, e.g., human IgG1, IgG2, IgG3, or IgG4, by at least one amino acid modification.

[0077] As used herein, "IgG subclass modification" or "isotype modification" refers to an amino acid modification that converts one amino acid of one IgG isotype to the corresponding amino acid of a different aligned IgG isotype. For example, because IgG1 contains tyrosine and IgG2 contains phenylalanine at EU296, a substitution of F296Y in IgG2 is considered an IgG subclass modification.

[0078] As used herein, "non-naturally occurring modification" refers to a non-isotypic amino acid modification. For example, the substitution 434S in IgG1, IgG2, IgG3, or IgG4 (or hybrids thereof) is considered a non-naturally occurring modification because none of the human IgGs contain serine at position 434.

[0079] As used herein, the terms "oligonucleotide" and "polynucleotide," used interchangeably, refer to a linear polymer of natural or modified nucleoside monomers linked by phosphodiester bonds or their analogs. The term "oligonucleotide" typically refers to shorter polymers, e.g., those containing about 3 to about 100 monomers, while the term "polynucleotide" typically refers to longer polymers, e.g., those containing about 100 monomers to several thousand monomers, e.g., 10,000 or more monomers. Oligonucleotides and polynucleotides can be natural or synthetic. Oligonucleotides and polynucleotides can include deoxyribonucleosides, ribonucleosides, and / or non-natural analogs thereof. In embodiments, oligonucleotides or polynucleotides can specifically bind to a target genome through regular patterns of monomer-monomer interactions, such as Watson-Crick base pairing, base stacking, Hoogsteen or reverse Hoogsteen base pairing, etc.

[0080] As used herein, "3E10 antibody" refers to an antibody identified according to the Kabat system, having a set of heavy chain CDRs (VH CDR1, VH CDR2, and VH CRD3) comprising amino acid sequences that differ by no more than two amino acids from SEQ ID NOs: 58, 59, and 60, respectively, and a set of light chain CDRs (VL CDR1, VL CDR2, and VL CRD3) comprising amino acid sequences that differ by no more than two amino acids from SEQ ID NOs: 61, 62, and 63, respectively, that binds to nucleic acid, and that is cell-permeable at least when bound to nucleic acid. As described herein, the 3E10 antigen is a polynucleotide.

[0081] As used herein, the term "cell-penetrating" refers to an antibody or antigen-binding fragment thereof that can penetrate cells, e.g., mammalian cells, without the aid of an exogenous delivery vehicle such as a liposome or a complexed cell-penetrating peptide. With respect to the 3E10 antibody and antigen-binding fragment thereof, the cell-penetrating antibody or antigen-binding fragment thereof can penetrate cells that express the ENT2 receptor on their cell surface, e.g., in the presence of a nucleic acid non-covalently bound and / or complexed to the 3E10 antibody or antigen-binding fragment thereof, resulting in internalization of the 3E10 antibody and antigen-binding fragment thereof. In some embodiments, the cell-penetrating 3E10 antibody or antigen-binding fragment thereof is complexed to a functional molecule, e.g., a chemical agent, a polynucleotide, or a polypeptide. Although cell-penetrating molecules are generally referred to herein as "cell-penetrating antibodies," antigen-binding fragments, variants, binding proteins, and fragments, including fusion proteins such as scFvs, di-scFvs, tri-scFvs, and other single-chain variable fragments, as well as other cell-penetrating molecules disclosed herein, are also expressly provided for use in the compositions, conjugates, and methods disclosed herein. Autoantibodies against double-stranded deoxyribonucleic acid (dsDNA) are frequently identified in the sera of systemic lupus erythematosus (SLE) patients and are often associated with the pathogenesis of the disease. Thus, in embodiments, cell-penetrating antibodies (e.g., cell-penetrating anti-DNA antibodies) can be derived from or isolated from SLE patients or animal models of SLE.

[0082] As used herein, "antibody-drug conjugate" or "ADC" refers to an antibody or antigen-binding fragment thereof or variant thereof covalently linked or conjugated to a bioactive molecule, e.g., a drug or anti-tumor agent.

[0083] As used herein, a "linker" refers to any chemical moiety capable of linking or connecting a compound, typically a drug or antitumor drug, to a cell-binding agent, such as an antibody, such as the 3E10 antibody or a fragment thereof, in a stable, covalent manner. In embodiments, a "linker" refers to any chemical moiety capable of linking or connecting a compound, such as a DNA damage-inducing agent, a DNA repair inhibitor, an immunomodulatory molecule, an alkylating agent, a microtubule inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, an adoptive cell therapy, or a topoisomerase inhibitor, to a cell-binding agent, such as the 3E10 antibody or a fragment thereof, in a stable, covalent manner. The linker can be sensitive to or substantially resistant to acid-induced cleavage, photo-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, and / or disulfide bond cleavage under conditions under which the compound and / or antibody remain active. Suitable linkers are well known in the art and include, for example, disulfide groups, thioether groups, acid-labile groups, photolabile groups, peptidase-labile groups, and esterase-labile groups. Linkers also include charged linkers, as described herein and known in the art, and hydrophilic forms thereof.

[0084] As used herein, the term "subject" refers to any individual who is the target of administration. A subject can be any animal (e.g., mammal), including humans and non-human animals (including, but not limited to, non-human primates, dogs, cats, rodents, horses, cows, pigs, mice, rats, hamsters, rabbits, etc.) (e.g., will be the recipient of a particular treatment). In an embodiment, the subject is a human.

[0085] In embodiments, the methods of the present disclosure are useful in treating a human subject. In embodiments, the human may be referred to as a patient. In embodiments, the human is female. In embodiments, the human is male. In embodiments, the human has an age ranging from about 1 to about 18 months old, about 18 to about 36 months old, about 1 to about 5 years old, about 5 to about 10 years old, about 10 to about 15 years old, about 15 to about 20 years old, about 20 to about 25 years old, about 25 to about 30 years old, about 30 to about 35 years old, about 35 to about 40 years old, about 40 to about 45 years old, about 45 to about 50 years old, about 50 to about 55 years old, about 55 to about 60 years old, about 60 to about 65 years old, about 65 to about 70 years old, about 70 to about 75 years old, about 75 to about 80 years old, about 80 to about 85 years old, about 85 to about 90 years old, about 90 to about 95 years old, or about 95 to about 100 years old.

[0086] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals in which a population of cells is characterized by unregulated cell growth. Examples of cancer include colon cancer, pancreatic cancer, lung cancer, ovarian cancer, liver cancer, breast cancer, brain cancer, kidney cancer, prostate cancer, gastrointestinal cancer, melanoma, cervical cancer, bladder cancer, glioblastoma, head and neck cancer, lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous B-cell lymphoma, cutaneous T-cell lymphoma, Waldenstrom's macroglobulinemia, chronic lymphocytic leukemia, leukemia, neuroblastoma, Werms' tumor, bone cancer, brain stem tumor, childhood diffuse intrinsic pontine glioma (DIPGL), and pediatric diffuse intrinsic pontine glioma (DGL). These include, but are not limited to, retinoblastoma, rhabdoid tumor, sarcoma, spinal cord tumor, endocrine carcinoma, esophageal carcinoma, gastric carcinoma, hepatobiliary carcinoma, myeloma, renal carcinoma, thyroid carcinoma, uterine carcinoma, carcinoma, blastoma, papilloma, adenoma, astrocytic tumor, oligodendroglial tumor, oligoastrocytic tumor, ependymal tumor, choroid plexus tumor, neuronal or mixed neuronal-glial tumor, pineal tumor, embryonal tumor, or neuroepithelial tumor not otherwise classified.

[0087] "Tumor" and "neoplasm" refer to a mass of tissue resulting from excessive cell growth or proliferation, whether benign (non-cancerous) or malignant (cancerous), and include precancerous lesions.

[0088] The terms "cancer cells," "tumor cells," and their grammatical equivalents refer to the total population of cells derived from a tumor or precancerous lesion, including both non-tumorigenic cells and tumorigenic stem cells (cancer stem cells), which make up the majority of the tumor cell population.

[0089] The term "chemotherapeutic warhead" refers to a cytotoxic or anti-tumor drug covalently attached to an antibody in an ADC.

[0090] As used herein, the term "pharmaceutically effective amount" means that the amount of the composition used is sufficient to ameliorate one or more causes or symptoms of a disease or disorder. Such an improvement need only reduce or alter, not necessarily eliminate, the exact dosage will vary depending on various factors, such as subject-dependent variables (e.g., age, immune system health, etc.), the disease or disorder being treated, and the route of administration and pharmacokinetics of the administered agent.

[0091] As used herein, the term "carrier" or "excipient" refers to an organic or inorganic ingredient, natural or synthetic, in a formulation with which one or more active ingredients are combined. In embodiments, the carrier or excipient is selected to minimize degradation of the active ingredient or to minimize adverse side effects in the subject, as would be known to one of skill in the art.

[0092] As used herein, the term "treating" refers to the medical management of a patient with the intent to cure, ameliorate, stabilize, or prevent a disease, pathological condition, or disorder. This term includes active treatment, i.e., treatment specifically directed at ameliorating a disease, pathological condition, or disorder, and also includes causal treatment, i.e., treatment directed at eliminating the cause of the associated disease, pathological condition, or disorder. In addition, this term includes palliative treatment, i.e., treatment designed to relieve symptoms rather than cure the disease, pathological condition, or disorder; preventative treatment, i.e., treatment aimed at minimizing or partially or completely inhibiting the onset of the associated disease, condition, or disorder; and supportive treatment, i.e., treatment used to complement another specific treatment aimed at ameliorating the associated disease, condition, or disorder.

[0093] A "pharmacologically effective amount," "pharmacologically effective dose," "therapeutically effective amount," or "effective amount" refers to an amount sufficient to produce a desired physiological effect or an amount capable of achieving a desired result, particularly for treating or preventing a disorder or disease. As used herein, an effective amount would include, for example, an amount sufficient to delay the onset of symptoms of a disorder or disease, alter the course of symptoms of a disorder or disease (e.g., slow the progression of symptoms of the disease), reduce or eliminate one or more symptoms or manifestations of a disorder or disease, and reverse symptoms of a disorder or disease. Therapeutic benefit also includes halting or slowing the progression of the underlying disease or disorder, regardless of whether improvement is achieved.

[0094] Effective doses, toxicity, and therapeutic effects can be determined by standard pharmaceutical procedures in cell cultures, tissue samples, tissue homogenates, or experimental animals to determine, for example, the LD50 (the dose lethal to approximately 50% of the population) and the ED50 (the dose therapeutically effective in approximately 50% of the population) or maximum tolerated dose. This dosage can vary depending on the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio LD50 / ED50. In embodiments, compositions, conjugates, and methods exhibiting large therapeutic indices are preferred. Therapeutically effective doses can be initially estimated from in vitro assays, including, for example, cell culture assays or measurements. A dose can also be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 as determined in cell culture or an appropriate animal model. Levels of the described compositions in plasma can be measured, for example, by high performance liquid chromatography. The effect of any particular dosage can be monitored by a suitable bioassay. Dosages can be determined by the physician and adjusted, if necessary, to suit the observed therapeutic effect.

[0095] In embodiments, the effect will result in a quantifiable change of at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 70%, or at least about 90%. In embodiments, the effect will result in a quantifiable change of about 10%, about 20%, about 30%, about 50%, about 70%, or even about 90%. Therapeutic benefit also includes halting or slowing the progression of the underlying disease or disorder, regardless of whether improvement is achieved.

[0096] III. Antibody-Drug Conjugates (ADCs) The present disclosure is directed, in part, to antibody drug conjugates (ADCs) comprising a cell-penetrating antibody, e.g., a 3E10 antibody or antigen-binding fragment thereof or variant thereof, conjugated via a linker to a biologically active molecule or therapeutic agent.

[0097] In embodiments, the ADC specifically binds to the epitope and causes cell death through the delivery of a drug or anti-tumor agent. In embodiments, the antibody-drug conjugate (ADC) has the formula A-(LP r ) q wherein A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is a linker, and P is a payload as described herein.

[0098] In embodiments, the present disclosure relates to the use of 3E10 antibodies and derivatives thereof for delivering therapeutic agents to a subject. While generally referred to herein as "3E10" or "3E10 antibodies," it will be understood that fragments, variants, and binding proteins, including antigen-binding fragments and fusion proteins such as scFv, di-scFv, tr-scFv, and other single-chain variable fragments, as well as other cell-permeable nucleic acid transport molecules disclosed herein, are encompassed by this term and are also expressly provided for use in the compositions, conjugates, and methods disclosed herein. Accordingly, antibodies and other binding proteins are also referred to herein as cell-permeable.

[0099] In embodiments, the antibody is conjugated to a therapeutic agent via a linker. In embodiments, the antibody is a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, as described herein. In embodiments, the antibody is a humanized 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, as described herein. Any type of agent, such as inorganic and organic molecules, pharmaceutical agents, drugs, peptides, proteins, genetic material, etc., can be delivered via conjugation with the 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, or a humanized 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, as described herein. In embodiments, the antibody-drug conjugate (ADC) comprises a drug or an anti-tumor drug.

[0100] A. Antigen-binding domain (ABD) As used herein, the term "antigen binding domain" or "ABD" refers to a domain comprising a three-dimensional structure capable of immunospecifically binding to an epitope of an antigen. Thus, in embodiments, the ABD comprises a hypervariable region of an antibody, optionally a VH and / or VL domain, optionally at least a VH domain. In embodiments, the ABD comprises at least one complementarity determining region (CDR) of an antibody. In embodiments, the ABD comprises at least two CDRs of an antibody. In embodiments, the ABD comprises at least three CDRs of an antibody. In embodiments, the ABD comprises at least four CDRs of an antibody. In embodiments, the ABD comprises at least five CDRs of an antibody. In embodiments, the ABD comprises at least six CDRs of an antibody.

[0101] 1.3E10 antibody In embodiments, the present disclosure relates to the use of 3E10 antibodies and derivatives thereof for delivering therapeutic agents to a subject. While generally referred to herein as "3E10" or "3E10 antibodies," it will be understood that fragments, variants, and binding proteins, including antigen-binding fragments and fusion proteins such as scFv, di-scFv, tr-scFv, and other single-chain variable fragments, as well as other cell-permeable nucleic acid transport molecules disclosed herein, are encompassed by this term and are also expressly provided for use in the compositions, conjugates, and methods disclosed herein. Accordingly, antibodies and other binding proteins are also referred to herein as cell-permeable.

[0102] In embodiments, the 3E10 antibody comprises the VL CDRs of SEQ ID NOs: 61, 62, and 63 and the VH CDRs of SEQ ID NOs: 58, 59, and 60. In embodiments, the 3E10 antibody comprises the VL CDRs of SEQ ID NOs: 9, 10, and 11 and the VH CDRs of SEQ ID NOs: 3, 4, and 5. In embodiments, the 3E10 antibody comprises the VL CDRs of SEQ ID NOs: 22, 23, and 24 and the VH CDRs of SEQ ID NOs: 15, 17, and 18. In embodiments, the 3E10 antibody comprises the VL CDRs of SEQ ID NOs: 9, 10, and 11 and the VH CDRs of SEQ ID NOs: 16, 4, and 5.

[0103] In embodiments, the 3E10 antibody has nucleic acid binding affinity.

[0104] In embodiments, the 3E10 antibody is competent for ENT2-mediated cellular internalization when bound to nucleic acids.

[0105] In embodiments, the targeting of the ADCs described herein, when used in methods for treating cancer or targeting cancer cells, tumors, or tumor cells, is not limited to tissue-specific antigens or cancer-specific antigens or tumor epitopes. Thus, the ADCs provided herein can broadly target cancers based on the cell surface expression of ENT2, a nucleoside transporter overexpressed in cancer and tumor cells. Advantageously, the ADCs described herein can simultaneously target ENT2 and extracellular DNA. Importantly, 3E10 has been shown to preferentially localize to the nuclei of tumor cells in vivo, likely due to increased DNA in the local environment released from ischemic and necrotic areas of the tumor. Targeting of the 3E10 antibody to extracellular DNA is described, for example, in Weisbart, Sci Reports, 2015, which is incorporated herein by reference. By targeting extracellular DNA as well as ENT2, the 3E10 ADCs described herein offer a platform for targeting various cancers and delivering chemotherapeutic agents to target and kill cancer cells.

[0106] In embodiments of the disclosure, the antibody or antigen-binding fragment or variant thereof is a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody.

[0107] In embodiments, the ADCs of the disclosure penetrate into cells and nuclei in an ENT2-dependent manner.

[0108] In embodiments, the ADC of the present disclosure further comprises a conjugated polynucleotide, and A is conjugated to the polynucleotide. In embodiments, the conjugated polynucleotide is pre-complexed with the ADC. In embodiments, the conjugated polynucleotide is an extracellular polynucleotide that is bound by the ADC at a site of interest, such as a site of ischemia and necrosis in a tumor. In embodiments, the polynucleotide is DNA. In embodiments, the polynucleotide is RNA.

[0109] In embodiments, the ABD comprises the VH domain and / or VL domain of the 3E10 antibody. In embodiments, the ABD comprises the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the 3E10 antibody.

[0110] In embodiments, the present disclosure provides a compound of formula A-(LP r ) qwherein A is a 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is a linker, and P is a payload as described herein, wherein the linker L connects A to P. In embodiments of the present disclosure, the amino acid residue corresponding to D31 in the heavy chain CDR1 of the 3E10 antibody or antigen-binding fragment thereof or variant thereof is substituted with N. It is known in the art that mutating the aspartic acid at residue 31 in VH CDR1 to asparagine increases the cationic charge of this residue, enhancing nucleic acid binding and delivery in vivo (3E10-D31N). In embodiments, additional 3E10 antibody variants include mutating the aspartic acid at residue 31 in VH CDR1 to arginine (3E10-D31R), which modeling shows an expanded cationic charge, or mutating it to lysine (3E10-D31K), which modeling shows an altered charge orientation. Thus, in embodiments, the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprises a D31R or D31K substitution. In embodiments, further 3E10 antibody variants comprise R96N and / or S30D substitutions, alone or in combination with D31N, D31R, or D31K. All sequences disclosed herein having residues corresponding to D31 or N31 of 3E10 are expressly disclosed with the D31R or D31K or N31R or N31K substitution.

[0111] In embodiments, the present disclosure provides a compound of formula A-(LP r ) qwherein A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is a linker, P is a payload described herein, r is an integer between 1 and 4, and q is an integer between 1 and 16, and the linker L connects A to (P); wherein the 3E10 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of X1YGMX2, where X1 is D, E, N, Q, R, or K, and X2 is K, R, or H (SEQ ID NO: 58). In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises: (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of X1ASX2X3VSTSSYSYX4X5, where X1 is K, R, or H, X2 is K, R, or H, X3 is T or S, X4 is M or L, and X5 is K, R, H, or A (SEQ ID NO: 61); (b) a VL CDR2 comprising the amino acid sequence of YASYLX1S, where X1 is D, E, N, or Q (SEQ ID NO: 62); (c) a VL CDR3 comprising the amino acid sequence of QX1SX2X3FPWT, where X1 is K, R, or H, X2 is K, R, or H, and X3 is D or E (SEQ ID NO: 63); and (d) a heavy chain variable region (VH) comprising the amino acid sequence of X1YGMX2. (e) a VH CDR2 comprising the amino acid sequence of YISSX1SSTIYYAX2X3VX4G (wherein X1 is G or S, X2 is D or E, X3 is T or S, and X4 is K, R, or H) (SEQ ID NO: 59); and (f) a VH CDR3 comprising the amino acid sequence of X1GLLLX2Y (wherein X1 is K, R, or H and X2 is D or E) (SEQ ID NO: 60).

[0112] In embodiments, the present disclosure provides antibody-drug conjugates (ADCs) having the formula A-(L-Pr)q, where A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is a linker, P is a payload described herein, r is an integer from 1 to 4, and q is an integer from 1 to 16, and the linker L connects A to (P), wherein the 3E10 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of NYGMH (SEQ ID NO: 15). In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises (a) a light chain variable region (VL) complementarity-determining region (CDR)1 comprising the amino acid sequence of RASKSVSTSSYSYMH (SEQ ID NO: 9), (b) a VL CDR2 comprising the amino acid sequence of YASYLES (SEQ ID NO: 10), (c) a VL CDR3 comprising the amino acid sequence of QHSREFPWT (SEQ ID NO: 11), (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of NYGMH (SEQ ID NO: 15), (e) a VH CDR2 comprising the amino acid sequence of YISSGSSTIYYADTVKG (SEQ ID NO: 4), and (f) a VH CDR3 comprising the amino acid sequence of RGLLLDY (SEQ ID NO: 5). In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises a light chain variable region (VL) comprising an amino acid sequence identical to SEQ ID NO: 21. In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises a heavy chain variable region (VH) comprising an amino acid sequence identical to SEQ ID NO: 14. In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises a full-length light chain (LC) comprising an amino acid sequence identical to SEQ ID NO: 20. In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof, comprises a full-length heavy chain (HC) comprising an amino acid sequence identical to SEQ ID NO: 13.

[0113] In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof comprises a light chain variable region (VL) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 21. In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof comprises a heavy chain variable region (VH) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 14. In embodiments, the antibody, or antigen-binding fragment thereof, or variant thereof comprises a full-length light chain (LC) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO: 20. In embodiments, the antibody or antigen-binding fragment or variant thereof comprises a full-length heavy chain (HC) comprising an amino acid sequence that is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical to SEQ ID NO:13.

[0114] In embodiments, the 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, can be transported into the cytoplasm and / or nucleus of a cell without the aid of a carrier or complex. For example, monoclonal antibody 3E10 and active fragments thereof that are transported in vivo to the nucleus of mammalian cells without cytotoxic effects are disclosed in U.S. Patent Nos. 4,812,397 and 7,189,396 to Richard Weisbart (each of which is incorporated by reference in its entirety).

[0115] The amino acid sequences of the 3E10 monoclonal antibody and its antigen-binding fragments are known in the art. Exemplary sequences of the heavy and light chains of 3E10 are provided herein below.

[0116] The murine form of the 3E10 antibody is described in Zack, et al., Immunology and Cell Biology, 72:513-520 (1994), which is incorporated herein by reference in its entirety.

[0117] Amino acid variants of the 3E10 antibody are also known in the art and are described, for example, in Zack, et al., J. Immunol., 157(5):2082-8 (1996). For example, amino acid 31 in CDR1 of the heavy chain variable region of 3E10 affects nucleic acid binding and the ability of the antibody to penetrate the nucleus. Substituting the "wild-type" aspartic acid with asparagine ("D31N" mutation) (e.g., compared to the original mouse antibody) improves the nucleic acid binding and nuclear penetration of the antibody compared to the "wild-type" mouse antibody. See, e.g., Zack, et al., Immunology and Cell Biology, 72:513-520 (1994); Weisbart, et al., J. Autoimmun., 11, 539-546 (1998); and Weisbart, Int. J. Oncol., 25, 1867-1873 (2004), which are incorporated by reference in their entireties.

[0118] The sequences of the 3E10 antibody and its antigen-binding fragments or variants thereof comprising the D31N substitution are disclosed herein. In embodiments, the 3E10 antibody and its antigen-binding fragments or variants thereof disclosed herein comprise the D31N substitution. In embodiments, other amino acids are substituted at position 31 in the 3E10 antibody and its antigen-binding fragments or variants thereof disclosed herein. For example, substitutions of D31R, D31K, or D31R are incorporated into embodiments of the present disclosure.

[0119] Other 3E10 light chain sequences are known in the art (see, e.g., Zack, et al., J. Immunol., 15;154(4):1987-94 (1995); GenBank: L16981-1 - Mouse Ig rearranged L-chain gene, partial cds; GenBank: AAA65681-1 - immunoglobulin light chain, partial [Mus musculus]).

[0120] The structural unit of a conventional antibody typically comprises a tetramer. Each tetramer typically consists of two pairs of identical polypeptide chains, each pair having one "light chain" (typically approximately 25 kDa in molecular weight) and one "heavy chain" (typically approximately 50-70 kDa in molecular weight). Human light chains are classified as kappa and lambda light chains. In embodiments, the antibodies disclosed herein are IgA, IgD, IgE, IgG, or IgM, including any subtype or isotype thereof. In embodiments, the antibodies disclosed herein are based on the IgG class. In embodiments, the antibodies disclosed herein are based on one of the IgG subclasses, including, but not limited to, IgG1, IgG2, IgG3, and IgG4. Generally, IgG1, IgG2, and IgG4 are more frequently used than IgG3. It should be noted that IgG1 has different allotypes with polymorphisms at 356 (D or E) and 358 (L or M), and in embodiments, the antibodies disclosed herein are based on IgG1 with D or E at position 356 and / or L or M at position 358.

[0121] Light chains generally contain two domains: a variable light chain domain (which contains the light chain CDRs and, together with the variable heavy chain domain, forms the Fv region) and a constant light chain region (often called CL or CK). Heavy chains contain a variable heavy chain domain and a constant domain, including CH1, which contains CH2-CH3, - optional hinge - Fc domain.

[0122] The hypervariable region of an antibody generally includes amino acid residues from about amino acid residues 24-34 (LCDR1; "L" indicates light chain), 50-56 (LCDR2), and 89-97 (LCDR3) in the light chain variable region, and amino acid residues from about amino acid residues 31-35B (HCDR1; "H" indicates heavy chain), 50-65 (HCDR2), and 95-102 (HCDR3) in the heavy chain variable region; Kabat et al., SEQUENCES OF PROTEINS OF IMMUNOLOGICAL INTEREST, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991) and / or these residues form hypervariable loops (e.g., residues 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3) in the light chain variable region, and residues 26-32 (HCDR1), 53-55 (HCDR2), and 96-101 (HCDR3) in the heavy chain variable region; Chothia and Lesk (1987), J. Mol. Biol. 196:901-917. Particular CDRs useful in the compositions, conjugates, and methods are described below.

[0123] As will be appreciated by those skilled in the art, the exact numbering and arrangement of the CDRs may vary among various numbering systems. However, it should be understood that disclosure of a variable heavy and / or variable light chain sequence includes disclosure of the associated (unique) CDRs. Thus, disclosure of each variable heavy chain region is a disclosure of the VH CDRs (e.g., VH CDR1, VH CDR2, and VH CDR3), and disclosure of each variable light chain region is a disclosure of the VL CDRs (e.g., VL CDR1, VL CDR2, and VL CDR3).

[0124] Throughout this disclosure, the Kabat numbering system is generally used when referring to residues in the variable domains (approximately residues 1-107 for light chain variable regions and residues 1-113 for heavy chain variable regions), with the EU numbering system (e.g., Kabat et al., supra (1991)) being used for the Fc region. In embodiments, the specification uses the IMGT system to define the complementarity determining regions (CDRs) provided herein.

[0125] The present disclosure provides a number of different CDR sets. In this case, a "complete CDR set" includes three variable light chain CDRs, e.g., VL CDR1, VL CDR2, and VL CDR3, and three variable heavy chain CDRs, e.g., VH CDR1, VH CDR2, and VH CDR3. These can be part of a larger variable light chain or variable heavy chain domain, respectively. Furthermore, as more fully outlined herein, the variable heavy and variable light domains can be on separate polypeptide chains, when heavy and light chains are used (e.g., when a Fab is used), or on a single polypeptide chain, in the case of an scFv sequence.

[0126] As described herein above, the present disclosure refers to various antibody domains of the 3E10 antibody or antigen-binding fragment thereof or variants thereof, including, but not limited to, the Fc domain, CH1 domain, CH2 domain, CH3 domain, hinge domain, heavy chain constant domain (CH1-hinge-Fc domain or CH1-hinge-CH2-CH3), variable heavy chain (VH) domain, variable light chain (VL) domain, light chain constant domain, Fab domain, and scFv domain.

[0127] 2. Humanized antibodies In embodiments, antibodies of the present disclosure comprise a heavy chain variable region derived from a particular germline heavy chain immunoglobulin gene and / or a light chain variable region derived from a particular germline light chain immunoglobulin gene. For example, such antibodies can be composed of or consist of a murine antibody, chimeric antibody, humanized antibody, or antigen-binding fragment or variant thereof, comprising a heavy or light chain variable region that is the "product" or "derived" of a particular germline sequence, e.g., the sequence of the 3E10 antibody. Human antibodies that are the "product" or "derived" of a human germline immunoglobulin sequence can be identified as such by comparing the amino acid sequence of the human antibody to the amino acid sequences of human germline immunoglobulins and selecting the human germline immunoglobulin sequence that most closely resembles the sequence of the human antibody (i.e., has the greatest percent identity) (using methods outlined herein). Human antibodies that are the "product" or "derived" of a particular human germline immunoglobulin sequence may contain amino acid differences from the germline sequence, e.g., due to naturally occurring somatic mutations or intentionally introduced site-specific mutations. However, a humanized antibody is typically at least about 90% identical to the amino acid sequence encoded by a human germline immunoglobulin gene and contains amino acid residues that identify the antibody as derived from a human sequence when compared to the germline immunoglobulin amino acid sequence of another species (e.g., a murine germline sequence). In certain cases, a humanized antibody can be at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identical in amino acid sequence to the amino acid sequence encoded by the germline immunoglobulin gene. Typically, a humanized antibody derived from a particular human germline sequence will not display more than 10-20 amino acid differences from the amino acid sequence encoded by the human germline immunoglobulin gene. In certain cases, a humanized antibody may display no more than 5, or no more than 4, or no more than 3, or no more than 2, or no more than 1 amino acid difference from the amino acid sequence encoded by the germline immunoglobulin gene.

[0128] In one embodiment, the parent antibody is affinity matured as known in the art. Humanization and affinity maturation can employ structure-based methods, such as those described in U.S. Ser. No. 11 / 004,590, incorporated herein by reference. Selection-based methods can be employed to humanize and affinity mature antibody variable regions, including, but not limited to, Wu et al., 1999, J. Mol. Biol. 294:151-162; Baca et al., 1997, J. Biol. Chem. 272(16):10678-10684; Rosok et al., 1996, J. Biol. Chem. 271(37):22611-22618; Rader et al., 1998, Proc. Natl. Acad. Sci. USA, 95:8910-8915; Krauss et al., 2003, Protein Engineering, 16(10):753-759, all of which are incorporated herein by reference. Other humanization methods may involve grafting only a portion of the CDRs, including, but not limited to, those described in USSN 09 / 810,510; Tan et al., 2002, J. Immunol. 169:1119-1125; De Pascalis et al., 2002, J. Immunol. 169:3076-3084, all of which are incorporated herein by reference.

[0129] 3. Fc variants In embodiments, one or more amino acid modifications can be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant can comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3, or IgG4 Fc region) that includes an amino acid modification (e.g., a substitution) at one or more amino acid positions.

[0130] In embodiments, Fc region variants possess some, but not all, effector functions, making them desirable candidates for applications where antibody half-life in vivo is important yet certain effector functions (e.g., complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be performed to confirm reduced / depleted CDC and / or ADCC activity. For example, Fc receptor (FcR) binding assays known in the art can be performed to confirm that the antibody lacks FcγR binding (and thus likely lacks ADCC activity) but retains FcRn binding ability. To assess complement activation, a CDC assay can be performed (see, e.g., Gazzano-Santoro et al., J. Immunol. Methods, 202:163 (1996); Cragg, MS et al., Blood, 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood, 103:2738-2743 (2004)). Determination of FcRn binding and in vivo clearance / half-life can also be performed using methods known in the art (see, e.g., Petkova, SB et al., Int'l. Immunol. 18(12):1759-1769 (2006)).

[0131] In embodiments, the antibodies provided herein can have reduced effector function and therefore can comprise one or more substitutions of Fc region residues 238, 265, 269, 270, 297, 327, and 329 (U.S. Patent No. 6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297, and 327, including the so-called "DANA" Fc mutant with substitutions of residues 265 and 297 to alanine (U.S. Patent No. 7,332,581).

[0132] In embodiments, the Fc region variants provided herein may have improved or diminished binding to FcRs (see, e.g., U.S. Patent No. 6,737,056, WO 2004 / 056312, and Shields et al., J. Biol. Chem. 9(2):6591-6604 (2001)).

[0133] In embodiments, the Fc region variants provided herein comprise an Fc region with one or more amino acid substitutions that improve ADCC, e.g., substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbering of residues).

[0134] In embodiments, the Fc region variants provided herein include changes that result in altered (i.e., either improved or decreased) C1q binding and / or complement dependent cytotoxicity (CDC), e.g., as described in U.S. Pat. No. 6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164:4178-4184 (2000).

[0135] In embodiments, the Fc region variants provided herein contain changes that result in increased half-life and improved binding to the neonatal Fc receptor (FcRn), the receptor responsible for the transfer of maternal IgG to the fetus (Guyer et al. J. Immunol. 117:587 (1976) and Kim et al. J. Immunol. 24:249 (1994)), e.g., as described in US 2005 / 0014934 A1 (Hinton et al.). These antibodies comprise an Fc region with one or more substitutions that improve binding of the Fc region to FcRn. Such Fc variants include those with a substitution at one or more of the following Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, ​​413, 424, or 434, e.g., a substitution at Fc region residue 434 (U.S. Patent No. 7,371,826).

[0136] In embodiments, the Fc region variants provided herein include "knobs-in-holes" or "skew" variants, which refer to amino acid manipulations that create steric effects that favor heterodimer formation and disfavor homodimer formation, as described in USSN 61 / 596,846, Ridgway et al, Protein Engineering 9(7):617 (1996); Atwell et al, J. Mol. Biol. 1997, 270:26; U.S. Pat. No. 8,216,805, all of which are incorporated herein by reference in their entireties.

[0137] In embodiments, the Fc region variants provided herein include changes described in Duncan & Winter, Nature, 322:738-40 (1988); U.S. Patent Nos. 5,648,260, 5,624,821, and WO 94 / 29351.

[0138] 4. Antibody fragment In embodiments, the antibody portion of the ADCs described herein comprises an antigen-binding fragment of the 3E10 antibody or variants thereof. In embodiments, the antigen-binding fragment retains the desired biological activity of the 3E10 antibody. In embodiments, the antigen-binding fragment retains at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, or at least about 95% of the desired biological activity of the 3E10 antibody. In embodiments, the antigen-binding fragment retains the ability of the antibody to bind to a target antigen, e.g., a nucleic acid, e.g., DNA. In embodiments, the antigen-binding fragment retains the ability of the antibody to bind to a cellular receptor, e.g., ENT2. In embodiments, the antigen-binding fragment retains the ability of the antibody to be internalized by a target cell.

[0139] In embodiments, the 3E10 antibody or antigen-binding fragment thereof comprises a single-chain fragment variable (scFv), tandem dual-scFv, (scFv)2, minibody, VHH, scFv-Fc, cross-Mab, dual variable domain immunoglobulin (DVD-Ig), single-chain tandem fragment variable (scTaFv), diabody, tandem diabody (TandAb), Fabsc, modular IgG-scFv, Fab, or F(ab')2.

[0140] In embodiments, the antigen-binding fragment of the 3E10 antibody or its variants includes a cross-Mab. In the cross-Mab format, complementary mutations are introduced into the heavy chain constant region of each arm to create a so-called "hole and knob," which results in preferential association between the different arms and the formation of a heterodimer rather than two homodimers of the same arm. The exact residues mutated in the heavy chain constant region of a cross-Mab bispecific antibody to form the "hole" and "knob" may vary depending on the specific design and optimization goals of the antibody. For more information regarding cross-Mab antibodies, see, for example, Huang, J., et al., Journal of Biological Chemistry, 294(50):19001-10 (2019), the disclosure of which is incorporated herein by reference in its entirety.

[0141] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a bivalent dual variable domain immunoglobulin (DVD-Ig) of the 3E10 antibody or antigen-binding fragment thereof. In the DVD-Ig format, each arm of the antibody comprises two VH / VL pairs. In some embodiments, one of the VH / VL pairs comprises the CDRs of the VH and VL of 3E10. For more information regarding DVD-Ig antibodies, see, e.g., Polson AG, et al., Journal of immunotherapy. 29(3):241-50 (2006) and U.S. Patent No. 7,612,181, the disclosures of which are incorporated herein by reference in their entireties.

[0142] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a single-chain variable fragment (scFv). As used herein, "single-chain Fv" or "scFv" refers to a VH domain covalently linked to a VL domain via a linker, e.g., an scFv linker as discussed herein, to form a contiguous protein chain. The scFv domains can be in either N-terminal to C-terminal configuration (i.e., VH-linker-VL or VL-linker-VH). In the sequences depicted in the sequence listing and figures herein, the order of the VH and VL domains is indicated in the name; for example, H.X_L.Y means that the sequence from N-terminal to C-terminal is VH-linker-VL, and L.Y_H.X means that the sequence from N-terminal to C-terminal is VL-linker-VH.

[0143] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a tandem dual scFv. A tandem dual scFv has two linearly linked scFv domains. In some embodiments, each scFv domain is derived from a different antibody, providing independent antigen-binding specificity. In some embodiments, one of the scFv domains comprises the VH and VL CDRs of 3E10. For more information on tandem dual scFvs, see, for example, Bossen C, et al., MAbs, 4(2):200-08 (2012), the disclosure of which is incorporated herein by reference in its entirety.

[0144] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a dimeric scFv antibody (scFv). Dimeric scFv antibodies have two scFv domains linked in a dimeric configuration. In some embodiments, each scFv domain is derived from a different antibody, providing independent antigen-binding specificity. In some embodiments, one of the scFv domains comprises the VH and VL CDRs of 3E10. For more information regarding dimeric scFv antibodies, see, for example, Llewellyn C, et al., Journal of Immunological Methods, 273(1-2):33-44 (2002), the disclosure of which is incorporated herein by reference in its entirety.

[0145] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises an scFv-Fc. As referred to herein, "scFv-Fc" refers to a polypeptide consisting of the heavy and light chain variable regions of an antibody joined by a linker, followed by an Fc polypeptide chain of the antibody, optionally the Fc region of a human IgG antibody, e.g., an IgG1, IgG2, IgG3, or IgG4 antibody.

[0146] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a single-chain tandem fragment variable (scTaFv) antibody. Single-chain tandem fragment variable (scTaFv) antibodies are a type of bispecific antibody consisting of two variable fragment (VH and VL) domains linked in tandem. In some embodiments, one of the variable fragment domains comprises the VH and VL CDRs of 3E10. For more information regarding scTaFv antibodies, see, e.g., Schramm C, et al., MAbs 5(3):442-49 (2013), the disclosure of which is incorporated herein by reference in its entirety.

[0147] In embodiments, the antigen-binding fragment of the 3E10 antibody or its variant comprises a VHH. As used herein, the term "VHH" refers to the variable domain of the heavy chain of a heavy-chain antibody. A VHH is a molecule capable of recognizing an antigen via a single domain and is the smallest unit of an antibody molecule discovered to date. In embodiments, a VHH can comprise one or more heavy-chain variable domains derived from a heavy-chain antibody, and the number of heavy-chain variable domains contained in a VHH is not limited.

[0148] In embodiments, the antigen-binding fragment of the 3E10 antibody or its variants comprises a diabody. As used herein, "diabody" refers to a bivalent antibody comprising two polypeptide chains, each of which is too short to allow pairing between the two domains on the same chain, and each domain pairs with a complementary domain on another polypeptide chain. (See, e.g., Holliger et al., 1993, Proc. Natl. Acad. Sci. USA, 90:6444-48 and Poljak et al., 1994, Structure, 2:1121-23.) When the two polypeptide chains of a diabody are identical, the resulting diabody has two identical antigen-binding sites. In embodiments, one of the antigen-binding domains of the diabody comprises the CDRs of the VH and VL of 3E10. For more information regarding diabodies, see, for example, Hoogenboom HR, et al., Trends Biotechnol., 21(12):553-57 (2003), the disclosure of which is incorporated herein by reference in its entirety. Diabodies with two different antigen-binding sites can be prepared using polypeptide chains of different sequences. Similarly, as used herein, "triabody" and "tetrabody" refer to antibodies that contain three and four polypeptide chains, respectively, forming three and four antigen-binding sites, which may be the same or different.

[0149] The term "minibody" is used to refer to an scFv-CH3 fusion protein that self-assembles into an 80 kDa bivalent dimer (scFv-CH3)2.

[0150] In embodiments, the antigen-binding fragment of the 3E10 antibody or its variants comprises a tandem diabody (TandAb). A tandem diabody has two antigen-binding domains (VH and VL) linked in tandem by a flexible peptide linker. In some embodiments, one of the antigen-binding domains comprises the CDRs of the VH and VL of 3E10. For more information regarding diabodies, see, for example, Sidelmann JG, et al., Mol. Immunol., 45(9):2597-607 (2008), the disclosure of which is incorporated herein by reference in its entirety.

[0151] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises a Fabsc. As used herein, an antibody molecule in the "Fabsc" format generally refers to a bispecific antibody molecule having a Fab fragment, generally comprising a hinge region at the C-terminus of the Fab fragment linked to the N-terminus of the CH2 domain, which in turn is linked to the N-terminus of the scFv fragment.

[0152] In embodiments, the antigen-binding fragment of the 3E10 antibody or its variants comprises an scFab. An scFab, also known as a single-chain fragment antigen-binding (Fab), is a type of antibody fragment that combines a variable heavy (VH) domain and a variable light (VL) domain into a single polypeptide chain connected by a peptide linker. The domain structure of a Fabsc includes variable domains from both the heavy and light chains (VH and VL) and a peptide linker connecting the two domains. In addition to the variable domains, a Fabsc also contains a constant domain (CL) from the light chain and a hinge region from the heavy chain. In some embodiments, one of the antigen-binding domains comprises the CDRs of the VH and VL of 3E10. For more information regarding Fabscs, see, for example, Kettner, C., et al., Frontiers in Immunology, 8(8):453 (2017), the disclosure of which is incorporated herein by reference in its entirety.

[0153] In embodiments, the antigen-binding fragment of the 3E10 antibody or variant thereof comprises an IgG-scFv. An IgG-scFv is an antibody in which an scFv is fused to the light chain or heavy chain of an IgG. In some embodiments, the scFv comprises the CDRs of the VH and VL of 3E10. In some embodiments, the IgG comprises the CDRs of the VH and VL of 3E10. In some embodiments, the antibody is a F(ab')2.

[0154] 5. Bispecific antibodies In embodiments, the 3E10 antibody and its antigen-binding fragments or variants thereof can be modified to improve their therapeutic potential. For example, in embodiments, a cell-permeable anti-DNA antibody is conjugated to another antibody specific for a second therapeutic target in the cytoplasm and / or nucleus of a target cell. For example, in embodiments, the cell-permeable 3E10 antibody is a bispecific antibody having a first heavy chain and a first light chain derived from 3E10 and a second heavy chain and a second light chain derived from a monoclonal antibody that specifically binds to the second therapeutic target.

[0155] Bispecific antibodies and other binding proteins having a first heavy chain and a first light chain derived from 3E10 and a second heavy chain and a second light chain derived from a monoclonal antibody that specifically binds to a second target are discussed in Weisbart, et al., Mol. Cancer Ther., 11(10):2169-73 (2012), and Weisbart, et al., Int. J. Oncology, 25:1113-8 (2004), and U.S. Patent Application Publication No. 2013 / 0266570, which are specifically incorporated by reference in their entireties. In embodiments, the second target is specific to a target cell type, tissue, organ, or the like. Thus, the second heavy chain and second light chain can serve as a targeting moiety that directs the complex to a target cell type, tissue, or organ. In embodiments, the second heavy chain and second light chain can target hematopoietic stem cells, CD34, or other target cells by targeting a receptor or ligand expressed on the cell type of interest. + In embodiments, the second heavy chain and the second light chain target thymocytes, spleen cells, or cancer cells.

[0156] Bispecific antibodies can be used to target cytotoxic agents or drugs to cells expressing a specific antigen. These antibodies have two binding sites directed to two different antigens or to two different epitopes on the same antigen. For example, in embodiments, bispecific antibodies can include one arm for binding to ENT2 and another arm for a second target. Bispecific antibody design can include designing various antibodies with multiple binding arms. Techniques for producing bispecific antibodies are common in the art (Millstein et al., 1983, Nature, 305:537-539; Brennan et al., 1985, Science, 229:81; Suresh et al., 1986, Methods in Enzymol. 121:120; Traunecker et al., 1991, EMBO J. 10:3655-3659; Shalaby et al., 1992, J. Exp. Med. 175:217-225; Kostelny et al., 1992, J. Immunol. 148:1547-1553; Gruber et al., 1994, J. Immunol. 152:5368; and U.S. Pat. No. 5,731,168). Antibodies with more than two valencies are also contemplated. For example, trispecific antibodies can be prepared (Tutt et al., J. Immunol. 147:60 (1991)). In embodiments, contemplated bispecific antibodies disclosed herein may be conjugated as bispecific ADCs.

[0157] Heteroconjugate antibodies are also within the scope of the present disclosure. Heteroconjugate antibodies are composed of two covalently linked antibodies. Such antibodies have been proposed, for example, to target immune system cells to unwanted cells (U.S. Pat. No. 4,676,980). It is contemplated that antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate. In embodiments, the contemplated heteroconjugate antibodies disclosed herein can be conjugated as heteroconjugate ADCs.

[0158] 6.3E10 array In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein comprises CDR sequences corresponding to the parent 3E10 antibody.

[0159] Thus, in embodiments, the 3E10 antibody, or variant thereof, or antigen-binding fragment thereof, comprises: (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of X1ASX2X3VSTSSYSYX4X5, where X1 is K, R, or H, X2 is K, R, or H, X3 is T or S, X4 is M or L, and X5 is K, R, H, or A (SEQ ID NO: 61); (b) a VL CDR2 comprising the amino acid sequence of YASYLX1S, where X1 is D, E, N, or Q (SEQ ID NO: 62); and (c) a VL CDR3 comprising the amino acid sequence of QX1SX2X3FPWT. (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of X1YGMX2 (wherein X1 is D, E, N, Q, R, or K, and X2 is K, R, or H) (SEQ ID NO: 58); (e) a VH CDR2 comprising the amino acid sequence of YISSX1SSTIYYAX2X3VX4G (wherein X1 is G or S, X2 is D or E, X3 is T or S, and X4 is K, R, or H) (SEQ ID NO: 59); and (f) a VH CDR3 comprising the amino acid sequence of X1GLLLX2Y (wherein X1 is K, R, or H, and X2 is D or E) (SEQ ID NO: 60).

[0160] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a light chain variable region (VL) complementarity-determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH), CDR1 comprising the amino acid sequence of 3E10-VH-CDR1 (SEQ ID NO: 3), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0161] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein comprises the CDR sequence of a mutant 3E10 antibody comprising a D31N amino acid substitution in VH CDR1. Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof comprises a light chain variable region (VL) complementarity-determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1_D31N (sequence number 22), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2_D31N (sequence number 23), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3_D31N (sequence number 24), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1_D31N (sequence number 15), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2_D31N (sequence number 17), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3_D31N (sequence number 18).

[0162] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein refers to CDR sequences corresponding to the parent 3E10 antibody, optionally comprising a D31N amino acid substitution in VH CDR1. Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof comprises a light chain variable region (VL) complementarity-determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1a (SEQ ID NO: 16), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0163] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein comprises CDR sequences corresponding to a parent 3E10 antibody with known amino acid substitutions in one or more CDRs. Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein comprises one or more amino acid substitutions compared to the CDR sequence of the parent 3E10 or 3E10-D31N variant selected from a G to S substitution at position 5 of VH CDR2, a T to S substitution at position 14 of VH CDR2, an S to T substitution at position 5 of VL CDR1, an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2.

[0164] Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.1 (SEQ ID NO: 26) or 3E10-VH-CDR2.2 (SEQ ID NO: 27). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDRs 1-3 and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDRs 1-3 and VH CDRs 1 and 3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDRs 1-3 and VH CDRs 1 and 3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody or compared to the 3E10-D31N variant.

[0165] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.1 (SEQ ID NO: 28) or 3E10-VL-CDR1.2 (SEQ ID NO: 29). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody or compared to the 3E10-D31N variant.

[0166] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.1 (SEQ ID NO: 30). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody or compared to the 3E10-D31N variant.

[0167] While some of the amino acid substitutions above are fairly conservative (e.g., S to T at position 5 of VL CDR1), others are for amino acids with very different properties, e.g., M to L at position 14 of VL CDR1, H to A at position 15 of VL CDR1, and E to Q at position 6 of VL CDR2. Without being bound by theory, this suggests that at least these positions within the 3E10 CDR framework are tolerant to other amino acid substitutions.

[0168] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.3 (SEQ ID NO: 31). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 with one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10-D31N variant, e.g., as described herein.

[0169] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.3 (SEQ ID NO: 32). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 with one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10-D31N variant, e.g., as described herein.

[0170] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.2 (SEQ ID NO: 33). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 with one or more amino acid substitutions relative to the CDRs of the parent 3E10 antibody or relative to the 3E10-D31N variant, e.g., as described herein.

[0171] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR1 comprising the amino acid sequence of 3E10-VH-CDR1.c1 (SEQ ID NO: 34), 3E10-VH-CDR1.c2 (SEQ ID NO: 35), 3E10-VH-CDR1.c3 (SEQ ID NO: 36), 3E10-VH-CDR1.c4 (SEQ ID NO: 37), or 3E10-VH-CDR1.c5 (SEQ ID NO: 38). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR2 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR2 and 3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0172] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.c1 (SEQ ID NO: 39), 3E10-VH-CDR2.c2 (SEQ ID NO: 40), or 3E10-VH-CDR2.c3 (SEQ ID NO: 41). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDRs 1-3 and VH CDRs 1 and 3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDRs 1-3 and VH CDRs 1 and 3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDRs 1-3 and VH CDRs 1 and 3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0173] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3.c1 (SEQ ID NO: 42), 3E10-VH-CDR3.c2 (SEQ ID NO: 43), or 3E10-VH-CDR3.c3 (SEQ ID NO: 44). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0174] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.c1 (SEQ ID NO: 45), 3E10-VL-CDR1.c2 (SEQ ID NO: 46), 3E10-VL-CDR1.c3 (SEQ ID NO: 47), 3E10-VL-CDR1.c4 (SEQ ID NO: 48), 3E10-VL-CDR1.c5 (SEQ ID NO: 49), or 3E10-VL-CDR1.c6 (SEQ ID NO: 50). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDRs 2 and 3 and VH CDRs 1-3 that have one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0175] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.c1 (SEQ ID NO: 51). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0176] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3.c1 (SEQ ID NO: 52), 3E10-VL-CDR3.c2 (SEQ ID NO: 53), 3E10-VL-CDR3.c3 (SEQ ID NO: 54), 3E10-VL-CDR3.c4 (SEQ ID NO: 55), 3E10-VL-CDR3.c5 (SEQ ID NO: 56), or 3E10-VL-CDR3.c6 (SEQ ID NO: 57). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 2 and VH CDR1-3 according to the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 2 and VH CDR1-3 according to the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR1 and 2 and VH CDR1-3 having one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0177] It is also contemplated that the 3E10 antibody or variant thereof, or antigen-binding fragment thereof as described herein, comprises any combination of the 3E10 CDR amino acid substitutions described above.

[0178] Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence of 3E10-VH-CDR1m (SEQ ID NO: 58). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR1-3 and VH CDR2 and 3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR1-3 and VH CDR2 and 3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0179] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO: 59). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0180] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3m (SEQ ID NO: 60). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1-3 and VH CDR1 and 2 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0181] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1m (SEQ ID NO: 61). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR2 and 3 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0182] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO: 62). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 3 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0183] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO: 63). In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 2 and VH CDR1-3 in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 2 and VH CDR1-3 in accordance with the 3E10-D31N variant. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, further comprises VL CDR1 and 2 and VH CDR1-3 with one or more amino acid substitutions compared to the CDRs of the parent 3E10 antibody, e.g., as described herein.

[0184] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1m (SEQ ID NO: 61), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO: 62), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO: 63), a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1m (SEQ ID NO: 58), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO: 59), and a VH CDR3 comprising the amino acid sequence of E10-VH-CDR3m (SEQ ID NO: 60).

[0185] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein refers to CDR sequences that have more than one amino acid substitution compared to the parent 3E10 antibody, which optionally contains a D31N amino acid substitution in VH CDR1. Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR1 (SEQ ID NO: 9); a VL CDR2 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR2 (SEQ ID NO: 10); a VL CDR3 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VL-CDR3 (SEQ ID NO: 11); a heavy chain variable region (VH) CDR1 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDR1a (SEQ ID NO: 16); a VH CDR2 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDR2 (SEQ ID NO: 4); and a VH CDR3 comprising an amino acid sequence having no more than one amino acid substitution relative to 3E10-VH-CDR3 (SEQ ID NO: 5).

[0186] In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof described herein refers to CDR sequences that have no more than one amino acid substitution compared to the parent 3E10 antibody, which optionally contains a D31N amino acid substitution in VH CDR1. Thus, in embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof, comprises a VL CDR1 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR1 (SEQ ID NO: 9); a VL CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR2 (SEQ ID NO: 10); a VL CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VL-CDR3 (SEQ ID NO: 11); a heavy chain variable region (VH) CDR1 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR1a (SEQ ID NO: 16); a VH CDR2 comprising an amino acid sequence having no more than two amino acid substitutions relative to 3E10-VH-CDR2 (SEQ ID NO: 4); and a VH CDR3 comprising an amino acid sequence having no more than two amino acid substitutions relative to E10-VH-CDR3 (SEQ ID NO: 5).

[0187] Other variants of the 3E10 antibody or its variants, or its antigen-binding fragments, are also known in the art, as disclosed, for example, in Zack, et al., J. Immunol., 157(5):2082-8 (1996). For example, amino acid 31 of the heavy chain variable region of 3E10 has been confirmed to affect the ability of the antibody and its fragments to penetrate the nucleus and bind to DNA. The D31N mutation in CDR1 penetrates the nucleus and binds to DNA with much higher efficiency than the original antibody (Zack, et al., Immunology and Cell Biology, 72:513-520 (1994); Weisbart, et al., J. Autoimmun., 11, 539-546 (1998); Weisbart, Int. J. Oncol., 25, 1867-1873 (2004)). In embodiments, the 3E10 antibody or antigen-binding fragment or variant thereof described herein has a D31N substitution.

[0188] The ADCs described herein can be prepared with any 3E10 antibody or antigenic fragment thereof, or any humanized 3E10 antibody or antigenic fragment thereof, disclosed in the prior art, see, for example, WO2015 / 106290, 2016 / 033324, WO2019 / 018426, and WO2019 / 018428, each of which is specifically incorporated herein by reference in its entirety.

[0189] In embodiments, the ADC comprises a humanized 3E10 antibody. Generally, a humanized antibody has one or more amino acid residues introduced into it from a source that is non-human. These non-human amino acid residues are often referred to as "import" residues, usually from an "import" variable domain. Antibody humanization techniques generally involve the use of recombinant DNA technology to manipulate the DNA sequence encoding one or more polypeptide chains of an antibody molecule.

[0190] In embodiments, the present disclosure provides humanized antibodies, or antigen-binding fragments thereof, that incorporate any combination of humanized VL and VH sequences disclosed herein, as well as VL and VH sequences that have sequence identity to the VH or VL sequences described herein, e.g., at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity thereto.

[0191] In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of any one of SEQ ID NOs: 1, 13, or 71-84. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 1. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 13. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 71. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 72. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 73. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 74. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 75. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 76. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 77. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of the heavy chain having the sequence of SEQ ID NO: 78.In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 79. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 80. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 81. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 82. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 83. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof, comprising one, two, or three CDRs of the heavy chain having the sequence of SEQ ID NO:84.

[0192] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VH having the sequence of any one of SEQ ID NOs: 2, 14, 64-70, and 103-112. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 2. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 14. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 64. In embodiments, the ADCs provided herein comprise a 3E10 VH or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 65. In embodiments, the ADCs provided herein comprise a 3E10 VH or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a heavy chain having the sequence of SEQ ID NO: 66. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 67. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 68. In embodiments, the ADCs provided herein comprise a 3E10 VH or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 69. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 70. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 103.In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 104. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 105. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 106. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 107. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 108. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 109. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 110. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 111. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, or three CDRs of a VH having the sequence of SEQ ID NO: 112.

[0193] In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a light chain having the sequence of any one of SEQ ID NOs: 7, 20, or 91-102. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 7. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 20. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 91. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 92. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 93. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 94. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 95. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 96. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 97. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of the light chain having the sequence of SEQ ID NO: 98.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 99. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 100. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 101. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a light chain having the sequence of SEQ ID NO: 102.

[0194] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of any one of SEQ ID NOs: 8, 21, 85-90, or 113-121. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 8. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 21. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 85. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 86. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 87. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 88. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 89. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 90. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or antigen-binding fragment thereof, or variant thereof, comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 113. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 114.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 115. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 116. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 117. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 118. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 119. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 120. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, or three CDRs of a VL having the sequence of SEQ ID NO: 121.

[0195] In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of any one of SEQ ID NOs: 122-137. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 122. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 123. In embodiments, the ADCs provided herein comprise a 3E10 antibody, or an antigen-binding fragment thereof, or a variant thereof, comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 124. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 125. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 126. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 127. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 128. In embodiments, the ADCs provided herein comprise the 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 129.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 130. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 131. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 132. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 133. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 134. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 135. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 136. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof comprising one, two, three, four, five, or six CDRs of an scFv having the sequence of SEQ ID NO: 137.

[0196] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99% identity to any one of SEQ ID NOs: 1, 13, or 71-84. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 1. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 13. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 71. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 72.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 73. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 74. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 75. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 76. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 77.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 78. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 79. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 80. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 81. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:82.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 83. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a heavy chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 84.

[0197] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: 7, 20, or 91-102. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 7. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 20. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 91. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:92.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 93. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 94. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 95. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 96. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:97.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 98. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:99. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 100. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 101. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a light chain sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 102.

[0198] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: 2, 14, 64-70, or 103-112. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 2. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 14. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 64. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:65.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 66. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 67. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 68. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 69. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:70.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 103. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 104. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 105. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 106. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 107.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 108. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 109. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 110. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 111. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VH sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 112.

[0199] In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to any one of SEQ ID NOs: 8, 21, 85-90, or 113-121. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:8. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 21. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 85. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:86.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 87. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 88. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 89. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO:90. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 113.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 114. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 115. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 116. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 117. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 118.In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 119. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 120. In embodiments, the ADCs provided herein comprise a 3E10 antibody or antigen-binding fragment thereof or variant thereof having a VL sequence having at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100% identity to SEQ ID NO: 121.

[0200] In embodiments, the ADCs provided herein comprise a light chain variable domain (3E10-VL) comprising an amino acid sequence at least about 97% identical to an amino acid sequence selected from the group consisting of 3E10-VL-h1 (SEQ ID NO: 85), 3E10-VL-h2 (SEQ ID NO: 86), 3E10-VL-h3 (SEQ ID NO: 87), 3E10-VL-h4 (SEQ ID NO: 88), 3E10-VL-h5 (SEQ ID NO: 89), and 3E10-VL-h6 (SEQ ID NO: 90); and a light chain variable domain (3E10-VH) comprising an amino acid sequence at least about 97% identical to an amino acid sequence selected from the group consisting of 3E10-VL-h1 (SEQ ID NO: 85), 3E10-VL-h2 (SEQ ID NO: 86), 3E10-VL-h3 (SEQ ID NO: 87), 3E10-VL-h4 (SEQ ID NO: 88), 3E10-VL-h5 (SEQ ID NO: 89), and 3E10-VL-h6 (SEQ ID NO: 90). and a heavy chain variable domain (3E10-VH) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-h1 (SEQ ID NO: 64), 3E10-VH-h2 (SEQ ID NO: 65), 3E10-VH-h3 (SEQ ID NO: 66), 3E10-VH-h4 (SEQ ID NO: 67), 3E10-VH-h5 (SEQ ID NO: 68), 3E10-VH-h6 (SEQ ID NO: 69), and 3E10-VH-h7 (SEQ ID NO: 70).

[0201] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h1 (SEQ ID NO: 85). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h1 (SEQ ID NO: 85). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h1 (SEQ ID NO: 85). In embodiments, the sequence of 3E10-VL is 3E10-VL-h1 (SEQ ID NO: 85).

[0202] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h2 (SEQ ID NO: 86). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h2 (SEQ ID NO: 86). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h2 (SEQ ID NO: 86). In embodiments, the sequence of 3E10-VL is 3E10-VL-h2 (SEQ ID NO: 86).

[0203] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h3 (SEQ ID NO: 87). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h3 (SEQ ID NO: 87). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h3 (SEQ ID NO: 87). In embodiments, the sequence of 3E10-VL is 3E10-VL-h3 (SEQ ID NO: 87).

[0204] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h4 (SEQ ID NO: 88). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h4 (SEQ ID NO: 88). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h4 (SEQ ID NO: 88). In embodiments, the sequence of 3E10-VL is 3E10-VL-h4 (SEQ ID NO: 88).

[0205] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h5 (SEQ ID NO: 89). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h5 (SEQ ID NO: 89). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h5 (SEQ ID NO: 89). In embodiments, the sequence of 3E10-VL is 3E10-VL-h5 (SEQ ID NO: 89).

[0206] In embodiments, the sequence of 3E10-VL is at least about 97% identical to 3E10-VL-h6 (SEQ ID NO: 90). In embodiments, the sequence of 3E10-VL is at least about 98% identical to 3E10-VL-h6 (SEQ ID NO: 90). In embodiments, the sequence of 3E10-VL is at least about 99% identical to 3E10-VL-h6 (SEQ ID NO: 90). In embodiments, the sequence of 3E10-VL is 3E10-VL-h6 (SEQ ID NO: 90).

[0207] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h1 (SEQ ID NO: 64). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h1 (SEQ ID NO: 64). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h1 (SEQ ID NO: 64). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h1 (SEQ ID NO: 64). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h1 (SEQ ID NO: 64). In embodiments, the sequence of 3E10-VH is 3E10-VH-h1 (SEQ ID NO: 64).

[0208] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h2 (SEQ ID NO: 65). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h2 (SEQ ID NO: 65). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h2 (SEQ ID NO: 65). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h2 (SEQ ID NO: 65). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h2 (SEQ ID NO: 65). In embodiments, the sequence of 3E10-VH is 3E10-VH-h2 (SEQ ID NO: 65).

[0209] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h3 (SEQ ID NO: 66). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h3 (SEQ ID NO: 66). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h3 (SEQ ID NO: 66). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h3 (SEQ ID NO: 66). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h3 (SEQ ID NO: 66). In embodiments, the sequence of 3E10-VH is 3E10-VH-h3 (SEQ ID NO: 66).

[0210] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h4 (SEQ ID NO: 67). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h4 (SEQ ID NO: 67). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h4 (SEQ ID NO: 67). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h4 (SEQ ID NO: 67). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h4 (SEQ ID NO: 67). In embodiments, the sequence of 3E10-VH is 3E10-VH-h4 (SEQ ID NO: 67).

[0211] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h5 (SEQ ID NO: 68). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h5 (SEQ ID NO: 68). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h5 (SEQ ID NO: 68). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h5 (SEQ ID NO: 68). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h5 (SEQ ID NO: 68). In embodiments, the sequence of 3E10-VH is 3E10-VH-h5 (SEQ ID NO: 68).

[0212] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h6 (SEQ ID NO: 69). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h6 (SEQ ID NO: 69). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h6 (SEQ ID NO: 69). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h6 (SEQ ID NO: 69). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h6 (SEQ ID NO: 69). In embodiments, the sequence of 3E10-VH is 3E10-VH-h6 (SEQ ID NO: 69).

[0213] In embodiments, the sequence of 3E10-VH is at least about 95% identical to 3E10-VH-h7 (SEQ ID NO: 70). In embodiments, the sequence of 3E10-VH is at least about 96% identical to 3E10-VH-h7 (SEQ ID NO: 70). In embodiments, the sequence of 3E10-VH is at least about 97% identical to 3E10-VH-h7 (SEQ ID NO: 70). In embodiments, the sequence of 3E10-VH is at least about 98% identical to 3E10-VH-h7 (SEQ ID NO: 70). In embodiments, the sequence of 3E10-VH is at least about 99% identical to 3E10-VH-h7 (SEQ ID NO: 70). In embodiments, the sequence of 3E10-VH is 3E10-VH-h7 (SEQ ID NO: 70).

[0214] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises an amino acid sequence that is at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-LC-h1m (SEQ ID NO: 91), 3E10-LC-h2m (SEQ ID NO: 92), 3E10-LC-h3m (SEQ ID NO: 93), 3E10-LC-h4m (SEQ ID NO: 94), 3E10-LC-h5m (SEQ ID NO: 95), and 3E10-LC-h6m (SEQ ID NO: 96). and a heavy chain (3E10-HC) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-HC-h1m (SEQ ID NO: 71), 3E10-HC-h2m (SEQ ID NO: 72), 3E10-HC-h3m (SEQ ID NO: 73), 3E10-HC-h4m (SEQ ID NO: 74), 3E10-HC-h5m (SEQ ID NO: 75), 3E10-HC-h6m (SEQ ID NO: 76), and 3E10-HC-h7m (SEQ ID NO: 77).

[0215] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h1m (SEQ ID NO: 91). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h1m (SEQ ID NO: 91). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h1m (SEQ ID NO: 91). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h1m (SEQ ID NO: 91). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h1m (SEQ ID NO: 91). In embodiments, the sequence of 3E10-LC is 3E10-LC-h1m (SEQ ID NO: 91).

[0216] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h2m (SEQ ID NO: 92). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h2m (SEQ ID NO: 92). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h2m (SEQ ID NO: 92). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h2m (SEQ ID NO: 92). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h2m (SEQ ID NO: 92). In embodiments, the sequence of 3E10-LC is 3E10-LC-h2m (SEQ ID NO: 92).

[0217] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h3m (SEQ ID NO: 93). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h3m (SEQ ID NO: 93). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h3m (SEQ ID NO: 93). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h3m (SEQ ID NO: 93). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h3m (SEQ ID NO: 93). In embodiments, the sequence of 3E10-LC is 3E10-LC-h3m (SEQ ID NO: 93).

[0218] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h4m (SEQ ID NO: 94). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h4m (SEQ ID NO: 94). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h4m (SEQ ID NO: 94). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h4m (SEQ ID NO: 94). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h4m (SEQ ID NO: 94). In embodiments, the sequence of 3E10-LC is 3E10-LC-h4m (SEQ ID NO: 94).

[0219] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h5m (SEQ ID NO: 95). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h5m (SEQ ID NO: 95). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h5m (SEQ ID NO: 95). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h5m (SEQ ID NO: 95). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h5m (SEQ ID NO: 95). In embodiments, the sequence of 3E10-LC is 3E10-LC-h5m (SEQ ID NO: 95).

[0220] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h6m (SEQ ID NO: 96). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h6m (SEQ ID NO: 96). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h6m (SEQ ID NO: 96). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h6m (SEQ ID NO: 96). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h6m (SEQ ID NO: 96). In embodiments, the sequence of 3E10-LC is 3E10-LC-h6m (SEQ ID NO: 96).

[0221] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h1m (SEQ ID NO:71). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h1m (SEQ ID NO:71). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h1m (SEQ ID NO:71). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h1m (SEQ ID NO:71). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h1m (SEQ ID NO:71). In embodiments, the sequence of 3E10-HC is 3E10-HC-h1m (SEQ ID NO:71).

[0222] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h2m (SEQ ID NO: 72). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h2m (SEQ ID NO: 72). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h2m (SEQ ID NO: 72). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h2m (SEQ ID NO: 72). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h2m (SEQ ID NO: 72). In embodiments, the sequence of 3E10-HC is 3E10-HC-h2m (SEQ ID NO: 72).

[0223] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h3m (SEQ ID NO: 73). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h3m (SEQ ID NO: 73). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h3m (SEQ ID NO: 73). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h3m (SEQ ID NO: 73). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h3m (SEQ ID NO: 73). In embodiments, the sequence of 3E10-HC is 3E10-HC-h3m (SEQ ID NO: 73).

[0224] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h4m (SEQ ID NO: 74). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h4m (SEQ ID NO: 74). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h4m (SEQ ID NO: 74). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h4m (SEQ ID NO: 74). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h4m (SEQ ID NO: 74). In embodiments, the sequence of 3E10-HC is 3E10-HC-h4m (SEQ ID NO: 74).

[0225] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h5m (SEQ ID NO: 75). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h5m (SEQ ID NO: 75). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h5m (SEQ ID NO: 75). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h5m (SEQ ID NO: 75). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h5m (SEQ ID NO: 75). In embodiments, the sequence of 3E10-HC is 3E10-HC-h5m (SEQ ID NO: 75).

[0226] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h6m (SEQ ID NO: 76). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h6m (SEQ ID NO: 76). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h6m (SEQ ID NO: 76). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h6m (SEQ ID NO: 76). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h6m (SEQ ID NO: 76). In embodiments, the sequence of 3E10-HC is 3E10-HC-h6m (SEQ ID NO: 76).

[0227] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h7m (SEQ ID NO: 77). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h7m (SEQ ID NO: 77). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h7m (SEQ ID NO: 77). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h7m (SEQ ID NO: 77). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h7m (SEQ ID NO: 77). In embodiments, the sequence of 3E10-HC is 3E10-HC-h7m (SEQ ID NO: 77).

[0228] In embodiments, the ADCs provided herein comprise a light chain (3E10-LC) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-LC-h1 (SEQ ID NO: 97), 3E10-LC-h2 (SEQ ID NO: 98), 3E10-LC-h3 (SEQ ID NO: 99), 3E10-LC-h4 (SEQ ID NO: 100), 3E10-LC-h5 (SEQ ID NO: 101), and 3E10-LC-h6 (SEQ ID NO: 102); and a light chain (3E10-HC-h1 (SEQ ID NO: 78), 3E10-HC-h2 (SEQ ID NO: 79), 3E10-HC-h3 (SEQ ID NO: 80), 3E10-HC-h4 (SEQ ID NO: 81), 3E10-HC-h5 (SEQ ID NO: 82), 3E10-HC-h6 (SEQ ID NO: 83), and 3E10-HC-h7 (SEQ ID NO: 84).

[0229] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h1 (SEQ ID NO: 97). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h1 (SEQ ID NO: 97). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h1 (SEQ ID NO: 97). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h1 (SEQ ID NO: 97). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h1 (SEQ ID NO: 97). In embodiments, the sequence of 3E10-LC is 3E10-LC-h1 (SEQ ID NO: 97).

[0230] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h2 (SEQ ID NO: 98). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h2 (SEQ ID NO: 98). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h2 (SEQ ID NO: 98). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h2 (SEQ ID NO: 98). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h2 (SEQ ID NO: 98). In embodiments, the sequence of 3E10-LC is 3E10-LC-h2 (SEQ ID NO: 98).

[0231] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h3 (SEQ ID NO: 99). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h3 (SEQ ID NO: 99). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h3 (SEQ ID NO: 99). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h3 (SEQ ID NO: 99). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h3 (SEQ ID NO: 99). In embodiments, the sequence of 3E10-LC is 3E10-LC-h3 (SEQ ID NO: 99).

[0232] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h4 (SEQ ID NO: 100). In embodiments, the sequence of 3E10-LC is 3E10-LC-h4 (SEQ ID NO: 100).

[0233] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h5 (SEQ ID NO: 101). In embodiments, the sequence of 3E10-LC is 3E10-LC-h5 (SEQ ID NO: 101).

[0234] In embodiments, the sequence of 3E10-LC is at least about 95% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of 3E10-LC is at least about 96% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of 3E10-LC is at least about 97% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of 3E10-LC is at least about 98% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of 3E10-LC is at least about 99% identical to 3E10-LC-h6 (SEQ ID NO: 102). In embodiments, the sequence of 3E10-LC is 3E10-LC-h6 (SEQ ID NO: 102).

[0235] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h1 (SEQ ID NO: 78). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h1 (SEQ ID NO: 78). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h1 (SEQ ID NO: 78). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h1 (SEQ ID NO: 78). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h1 (SEQ ID NO: 78). In embodiments, the sequence of 3E10-HC is 3E10-HC-h1 (SEQ ID NO: 78).

[0236] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h2 (SEQ ID NO: 79). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h2 (SEQ ID NO: 79). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h2 (SEQ ID NO: 79). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h2 (SEQ ID NO: 79). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h2 (SEQ ID NO: 79). In embodiments, the sequence of 3E10-HC is 3E10-HC-h2 (SEQ ID NO: 79).

[0237] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h3 (SEQ ID NO: 80). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h3 (SEQ ID NO: 80). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h3 (SEQ ID NO: 80). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h3 (SEQ ID NO: 80). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h3 (SEQ ID NO: 80). In embodiments, the sequence of 3E10-HC is 3E10-HC-h3 (SEQ ID NO: 80).

[0238] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h4 (SEQ ID NO: 81). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h4 (SEQ ID NO: 81). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h4 (SEQ ID NO: 81). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h4 (SEQ ID NO: 81). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h4 (SEQ ID NO: 81). In embodiments, the sequence of 3E10-HC is 3E10-HC-h4 (SEQ ID NO: 81).

[0239] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h5 (SEQ ID NO: 82). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h5 (SEQ ID NO: 82). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h5 (SEQ ID NO: 82). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h5 (SEQ ID NO: 82). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h5 (SEQ ID NO: 82). In embodiments, the sequence of 3E10-HC is 3E10-HC-h5 (SEQ ID NO: 82).

[0240] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h6 (SEQ ID NO: 83). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h6 (SEQ ID NO: 83). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h6 (SEQ ID NO: 83). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h6 (SEQ ID NO: 83). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h6 (SEQ ID NO: 83). In embodiments, the sequence of 3E10-HC is 3E10-HC-h6 (SEQ ID NO: 83).

[0241] In embodiments, the sequence of 3E10-HC is at least about 95% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of 3E10-HC is at least about 96% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of 3E10-HC is at least about 97% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of 3E10-HC is at least about 98% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of 3E10-HC is at least about 99% identical to 3E10-HC-h7 (SEQ ID NO: 84). In embodiments, the sequence of 3E10-HC is 3E10-HC-h7 (SEQ ID NO: 84).

[0242] In embodiments, ADCs comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein have CDR sequences corresponding to the CDRs in the parent 3E10 antibody, optionally containing a D31N amino acid substitution in VH CDR1. Thus, in embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) complementarity-determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9), a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10), a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11), a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1a (SEQ ID NO: 16), a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4), and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0243] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises CDR sequences derived from a variant humanized 3E10 antibody comprising a D31N amino acid substitution in VH CDR1 (SEQ ID NO: 15).

[0244] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than seven amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0245] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have 10 or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0246] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have nine or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0247] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have eight or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0248] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than seven amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0249] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have six or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0250] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have five or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0251] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have four or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0252] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than three amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0253] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than two amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0254] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than one amino acid substitution compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), 3E10-VL-CDR3 (SEQ ID NO: 11), 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0255] Thus, in embodiments, the ADCs described herein may comprise a humanized 3E10 antibody or antigen-binding fragment thereof comprising: a light chain variable domain (VL) complementarity-determining region (CDR) 1 comprising the amino acid sequence of 3E10-VL-CDR1 (SEQ ID NO: 9); a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2 (SEQ ID NO: 10); a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3 (SEQ ID NO: 11); a heavy chain variable domain (VH) CDR1 comprising the amino acid sequence of 3E10-VH-CDR1_D31N (SEQ ID NO: 15); a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2 (SEQ ID NO: 4); and a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3 (SEQ ID NO: 5).

[0256] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a set of complementarity determining regions (CDRs) that collectively have no more than 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid substitutions compared to the CDR sequence of the 3E10-D31 variant (SEQ ID NOs: 15-18 and 22-24), selected from, but not limited to, a G to S substitution at position 5 of VH CDR2, a T to S substitution at position 14 of VH CDR2, an S to T substitution at position 5 of VL CDR1, an M to L substitution at position 14 of VL CDR1, an H to A substitution at position 15 of VL CDR1, and an E to Q substitution at position 6 of VL CDR2.

[0257] Thus, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.1 (SEQ ID NO: 26) or 3E10-VH-CDR2.2 (SEQ ID NO: 27). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 9-11) and VH CDRs 1 and 3 (SEQ ID NOs: 3 and 5) in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 22-24) and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) in accordance with the 3E10-D31N mutant.

[0258] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.1 (SEQ ID NO: 28) or 3E10-VL-CDR1.2 (SEQ ID NO: 29). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 10 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 23 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0259] Similarly, in embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.1 (SEQ ID NO: 30). In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 9 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 22 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0260] While some of the amino acid substitutions described above are fairly conservative (e.g., S to T at position 5 of VL CDR1), others are substitutions for amino acids with significantly different properties, e.g., M to L at position 14 of VL CDR1, H to A at position 15 of VL CDR1, and E to Q at position 6 of VL CDR2. Without being bound by theory, this suggests that at least these positions within the 3E10 CDR framework are tolerant to other amino acid substitutions.

[0261] Thus, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.3 (SEQ ID NO: 31). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 9-11) and VH CDRs 1 and 3 (SEQ ID NOs: 3 and 5) in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 22-24) and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) in accordance with 3E10-D31N.

[0262] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.3 (SEQ ID NO: 32). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 10 and 11), and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18), in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 23 and 24), and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18), in accordance with the 3E10-D31N variant.

[0263] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.2 (SEQ ID NO: 33). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 9 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 22 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0264] Thus, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence of 3E10-VH-CDR1.c1 (SEQ ID NO: 34), 3E10-VH-CDR1.c2 (SEQ ID NO: 35), 3E10-VH-CDR1.c3 (SEQ ID NO: 36), 3E10-VH-CDR1.c4 (SEQ ID NO: 37), or 3E10-VH-CDR1.c5 (SEQ ID NO: 38). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1-3 (SEQ ID NOs: 9-11), and VH CDR2 and 3 (SEQ ID NOs: 4 and 5) in accordance with the parent 3E10 antibody.

[0265] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2.c1 (SEQ ID NO: 39), 3E10-VH-CDR2.c2 (SEQ ID NO: 40), or 3E10-VH-CDR2.c3 (SEQ ID NO: 41). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 9-11) and VH CDRs 1 and 3 (SEQ ID NOs: 3 and 5) in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 22-24) and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) in accordance with the 3E10-D31N mutant.

[0266] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3.c1 (SEQ ID NO: 42), 3E10-VH-CDR3.c2 (SEQ ID NO: 43), or 3E10-VH-CDR3.c3 (SEQ ID NO: 44). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1-3 (SEQ ID NOs: 9-11) and VH CDR1 and 2 (SEQ ID NOs: 3 and 4) in accordance with the parent 3E10 antibody. In embodiments, the 3E10 antibody or variant thereof, or antigen-binding fragment thereof further comprises VL CDR1-3 (SEQ ID NOs: 22-24) and VH CDR1 and 2 (SEQ ID NOs: 15 and 17) in accordance with the 3E10-D31N variant.

[0267] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1.c1 (SEQ ID NO: 45), 3E10-VL-CDR1.c2 (SEQ ID NO: 46), 3E10-VL-CDR1.c3 (SEQ ID NO: 47), 3E10-VL-CDR1.c4 (SEQ ID NO: 48), 3E10-VL-CDR1.c5 (SEQ ID NO: 49), or 3E10-VL-CDR1.c6 (SEQ ID NO: 50). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 10 and 11), and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 2 and 3 (SEQ ID NOs: 23 and 24), and VH CDRs 1-3 (SEQ ID NOs: 15, 17, and 18) according to the 3E10-D31N variant.

[0268] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2.c1 (SEQ ID NO: 51). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 9 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 22 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0269] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3.c1 (SEQ ID NO: 52), 3E10-VL-CDR3.c2 (SEQ ID NO: 53), 3E10-VL-CDR3.c3 (SEQ ID NO: 54), 3E10-VL-CDR3.c4 (SEQ ID NO: 55), 3E10-VL-CDR3.c5 (SEQ ID NO: 56), or 3E10-VL-CDR3.c6 (SEQ ID NO: 57). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 2 (SEQ ID NOs: 9 and 10), and VH CDR1-3 (SEQ ID NOs: 3-5) according to the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 2 (SEQ ID NOs: 22 and 23), and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) according to the 3E10-D31N variant.

[0270] It is also contemplated that ADCs comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein may contain no more than 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 of the above-described CDR amino acid substitutions. Further examples of 3E10 mutant CDR sequences are described herein (SEQ ID NOs: 58-63).

[0271] Thus, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR1 comprising the amino acid sequence of 3E10-VH-CDR1m (SEQ ID NO: 58). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1-3 (SEQ ID NOs: 9-11), and VH CDR2 and 3 (SEQ ID NOs: 4 and 5) in accordance with the parent 3E10 antibody.

[0272] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR2 comprising the amino acid sequence of 3E10-VH-CDR2m (SEQ ID NO: 59). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 9-11) and VH CDRs 1 and 3 (SEQ ID NOs: 3 and 5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 22-24) and VH CDRs 1 and 3 (SEQ ID NOs: 15 and 18) in accordance with the 3E10-D31N variant.

[0273] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VH CDR3 comprising the amino acid sequence of 3E10-VH-CDR3m (SEQ ID NO: 60). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 9-11) and VH CDRs 1 and 2 (SEQ ID NOs: 3 and 4) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDRs 1-3 (SEQ ID NOs: 22-24) and VH CDRs 1 and 2 (SEQ ID NOs: 15 and 17) in accordance with the 3E10-D31N variant.

[0274] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR1 comprising the amino acid sequence of 3E10-VL-CDR1m (SEQ ID NO: 61). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 10 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR2 and 3 (SEQ ID NOs: 23 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0275] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR2 comprising the amino acid sequence of 3E10-VL-CDR2m (SEQ ID NO: 62). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 9 and 11) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 3 (SEQ ID NOs: 22 and 24) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0276] Similarly, in embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a VL CDR3 comprising the amino acid sequence of 3E10-VL-CDR3m (SEQ ID NO: 63). In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 2 (SEQ ID NOs: 9 and 10) and VH CDR1-3 (SEQ ID NOs: 3-5) in accordance with the parent 3E10 antibody. In embodiments, the humanized 3E10 antibody or antigen-binding fragment thereof further comprises VL CDR1 and 2 (SEQ ID NOs: 9 and 10) and VH CDR1-3 (SEQ ID NOs: 15, 17, and 18) in accordance with the 3E10-D31N variant.

[0277] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises a light chain variable domain comprising an amino acid sequence that is at least about 90% identical to an amino acid sequence selected from the group consisting of 3E10-VL-h1 (SEQ ID NO: 85), 3E10-VL-h2 (SEQ ID NO: 86), 3E10-VL-h3 (SEQ ID NO: 87), 3E10-VL-h4 (SEQ ID NO: 88), 3E10-VL-h5 (SEQ ID NO: 89), and 3E10-VL-h6 (SEQ ID NO: 90). and a light chain variable domain (3E10-VL), wherein the light chain variable domain (3E10-VL) further comprises one or more amino acid residues selected from proline (Pro) at position 15, threonine (Thr) at position 22, tyrosine (Tyr) at position 49, Thr at position 74, asparagine (Asn) at position 76, alanine (Ala) at position 80, Asn at position 81, Thr at position 83, Asn at position 85, and valine (Val) at position 104 of 3E10-VL according to the Kabat numbering system. The set of CDRs has collectively no more than six amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11), and the antibody comprises a set of 3E10-VL CDRs that collectively have no more than six amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0278] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs having five or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0279] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs having four or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0280] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs that have no more than three amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0281] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs having no more than two amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0282] In embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs that have no more than one amino acid substitution compared to the set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0283] In embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VL CDRs comprising a set of CDRs having the amino acid sequences of 3E10-VL-CDR1 (SEQ ID NO: 9), 3E10-VL-CDR2 (SEQ ID NO: 10), and 3E10-VL-CDR3 (SEQ ID NO: 11).

[0284] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a lysine (Lys) residue at position 49 of 3E10-VL according to Kabat numbering.

[0285] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a glutamic acid (Glu) residue at position 81 of 3E10-VL according to Kabat numbering.

[0286] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a proline (Pro) residue at position 15 of 3E10-VL according to Kabat numbering.

[0287] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a valine (Val) residue at position 104 of 3E10-VL according to Kabat numbering.

[0288] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof described herein comprises an ADC comprising an amino acid sequence at least about 90% identical to an amino acid sequence selected from the group consisting of 3E10-VH-h1 (SEQ ID NO: 64), 3E10-VH-h2 (SEQ ID NO: 65), 3E10-VH-h3 (SEQ ID NO: 66), 3E10-VH-h4 (SEQ ID NO: 67), 3E10-VH-h5 (SEQ ID NO: 68), 3E10-VH-h6 (SEQ ID NO: 69), and 3E10-VH-h7 (SEQ ID NO: 70). and a heavy chain variable domain (3E10-H) comprising the following amino acids according to the Kabat numbering of 3E10-VH: glutamine (Gln) at position 13, leucine (Leu) at position 18, arginine (Arg) at position 19, glycine (Gly) at position 42, serine (Ser) at position 49, Ser at position 77, tyrosine (Tyr) at position 79, Asn at position 82, Ala at position 84, Val at position 89, leucine (Leu) at position 108, Val at position 109, and Ser at position 113. and the antibody comprises a set of 3E10-VH CDRs that collectively have six or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0289] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that have five or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0290] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that have four or fewer amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0291] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that have no more than three amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0292] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that have no more than two amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0293] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that have no more than one amino acid substitution compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0294] In embodiments, an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof comprises a set of 3E10-VH CDRs that contain no more than 5, 4, 3, 2, or 1 amino acid substitutions compared to the set of CDRs having the amino acid sequences of 3E10-VH-CDR1_D31N (SEQ ID NO: 15), 3E10-VH-CDR2 (SEQ ID NO: 4), and 3E10-VH-CDR3 (SEQ ID NO: 5).

[0295] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with an arginine (Arg) residue at position 18 of 3E10-VH according to Kabat numbering.

[0296] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a (Lys) residue at position 19 of 3E10-VH according to Kabat numbering.

[0297] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with an alanine (Ala) residue at position 49 of 3E10-VH according to Kabat numbering.

[0298] In one aspect, the present disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a glutamine (Gln) residue at position 13 of 3E10-VH according to Kabat numbering.

[0299] In one aspect, the present disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a leucine (Leu) residue at position 108 of 3E10-VH according to Kabat numbering.

[0300] In one aspect, the disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a valine (Val) residue at position 109 of 3E10-VH according to Kabat numbering.

[0301] In one aspect, the present disclosure provides an ADC comprising a humanized antibody or antigen-binding fragment thereof with a serine (Ser) residue at position 113 of 3E10-VH according to Kabat numbering.

[0302] In embodiments, the present disclosure provides ADCs comprising a humanized 3E10 antibody or antigen-binding fragment thereof with a fragment crystallizable (Fc) region.

[0303] In embodiments, the present disclosure provides an ADC comprising a humanized 3E10 antibody or antigen-binding fragment thereof having an Fc region selected from human IgG1 Fc, human IgG2a Fc, human IgG2b Fc, human IgG3 Fc, and human IgG4 Fc.

[0304] In embodiments, the present disclosure provides an ADC comprising a humanized 3E10 antibody or variant thereof, or an antigen-binding fragment thereof, comprising a heavy chain constant domain (CH).

[0305] In embodiments, an ADC comprising a humanized 3E10 antibody or variant thereof, or an antigen-binding fragment thereof comprises an Fc region selected from human γ1 CH1, human γ2 CH1, human γ3 CH1, and human γ4 CH1.

[0306] In embodiments, the present disclosure provides an ADC comprising a humanized 3E10 antibody or variant thereof, or an antigen-binding fragment thereof, comprising the light chain constant domain (CL).

[0307] In one aspect, the present disclosure provides an ADC comprising a humanized 3E10 antibody or variant thereof comprising an Fc region selected from the group consisting of human λCL and human κCL.

[0308] In embodiments, the ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof may be selected from the group consisting of 3E10-VL-h1 (SEQ ID NO: 85) and 3E10-VH-h1 (SEQ ID NO: 64), 3E10-VL-h1 (SEQ ID NO: 85) and 3E10-VH-h2 (SEQ ID NO: 65), 3E10-VL-h1 (SEQ ID NO: 85) and 3E10-VH-h3 (SEQ ID NO: 66), 3E10-VL-h1 (SEQ ID NO: 85) and 3E10-VH-h4 (SEQ ID NO: 67), 3E10-VL-h2 (SEQ ID NO: 86) and 3E10-VH-h1 (SEQ ID NO: 64), 3E10-VL-h2 (SEQ ID NO: 86) and 3E10-VH-h2 (SEQ ID NO: 86) and 3E10-VH-h1 (SEQ ID NO: 64), and 3E10-VL-h2 (SEQ ID NO: 86) and 3E10-VH-h3 (SEQ ID NO: 66). The combinations include light chain variable domains (VL) and heavy chain variable domains (VH) selected from 3E10-VH-h2 (SEQ ID NO: 65), 3E10-VL-h3 (SEQ ID NO: 87) and 3E10-VH-h1 (SEQ ID NO: 64), 3E10-VL-h5 (SEQ ID NO: 89) and 3E10-VH-h5 (SEQ ID NO: 68), 3E10-VL-h5 (SEQ ID NO: 89) and 3E10-VH-h6 (SEQ ID NO: 69), 3E10-VL-h6 (SEQ ID NO: 90) and 3E10-VH-h5 (SEQ ID NO: 68), and 3E10-VL-h6 (SEQ ID NO: 90) and 3E10-VH-h6 (SEQ ID NO: 69).

[0309] In embodiments, an ADC comprising the humanized 3E10 antibody or antigen-binding fragment thereof comprises a combination of the light chain variable domain (VL) of 3E10-VL-H6 (SEQ ID NO: 90) and the heavy chain variable domain (VH) of 3E10-VH-H6 (SEQ ID NO: 69).

[0310] Antibodies useful in the compositions, conjugates, and methods described herein include whole immunoglobulins of any class (i.e., intact antibodies), fragments thereof containing at least the antigen-binding variable domain of an antibody, and synthetic proteins. The variable domains vary in sequence among antibodies and are responsible for the binding and specificity of each particular antibody for a particular antigen. However, diversity is not usually uniformly distributed within the variable domains of antibodies. Diversity is typically concentrated in three segments called complementarity-determining regions (CDRs) or hypervariable regions in both the light and heavy chain variable domains. The more highly conserved portions of the variable domains are called framework regions (FRs). Naturally occurring heavy and light chain variable domains each contain four FR regions that largely adopt a beta-sheet configuration, linked by three CDRs that form loops that connect, and in some cases form part of, the beta-sheet structure. The CDRs within each chain are held together in close proximity by the FR regions and, together with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies. Thus, antibodies typically contain at least the CDRs necessary to maintain DNA binding and / or interfere with DNA repair.

[0311] The 3E10 antibody is typically monoclonal 3E10, or a variant, derivative, fragment, fusion, or humanized version thereof that binds to the same or different epitope(s) as 3E10.

[0312] A deposit under the terms of the Budapest Treaty of a hybridoma cell line producing monoclonal antibody 3E10 was received on September 6, 2000, accepted by the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, VA 20110-2209, and assigned U.S. Patent Deposit No. PTA-2439. Thus, the antibody can have the same or a different epitope specificity as monoclonal antibody 3E10 produced by ATCC No. PTA2439 hybridoma. The antibody can have the paratope of monoclonal antibody 3E10. The antibody can be a single-chain variable fragment of 3E10, or a variant thereof, e.g., a conservative variant thereof. The antibody can be a single-chain variable fragment of 3E10 (3E10Fv), or a variant thereof.

[0313] Additionally or alternatively, the heavy chain complementarity determining regions (CDRs) can be defined according to the IMGT system. The complementarity determining regions (CDRs) identified by the IMGT system include CDR H1.3 (original sequence): GFTFSDYG (SEQ ID NO: 117); CDR H1.4 (with D31N mutation): GFTFSNYG (SEQ ID NO: 118); CDR H2.2: ISSGSSTI (SEQ ID NO: 119) and mutant ISSSSTI (SEQ ID NO: 120); CDR H3.2: ARGLLLDY (SEQ ID NO: 121).

[0314] Additionally or alternatively, the light chain complementarity determining regions (CDRs) can be defined according to the IMGT system, which identifies CDRs as follows: CDRL1.2: KSVSTSSYSY (SEQ ID NO: 122) and variant KTVSTSSYSY (SEQ ID NO: 123); CDRL2.2: YAS (SEQ ID NO: 124); CDRL3.2: QHSREFPWT (SEQ ID NO: 125).

[0315] The disclosed compositions, conjugates, and methods typically utilize antibodies that maintain the ability to penetrate cells and, optionally, the nucleus. The mechanisms of cellular internalization by autoantibodies are diverse. Some are internalized by electrostatic interactions or FcR-mediated endocytosis, while others utilize mechanisms based on association with cell surface myosin or calreticulin, followed by endocytosis (Ying-Chyi et al., Eur. J. Immunol. 38, 3178-3190 (2008); Yanase et al., J. Clin. Invest. 100, 25-31 (1997)). The 3E10 antibody and its antigen-binding fragments or variants thereof can cross cell membranes via equilibrative nucleoside (ENT) transporters. In embodiments, 3E10 crosses the cell membrane via the ENT1, ENT2, ENT3, or ENT4 transporter (see, e.g., WO2015 / 106290A1 and WO2016 / 033324A1, each of which is incorporated by reference in its entirety). In embodiments, 3E10 penetrates cells via an Fc-independent mechanism (as evidenced by the ability of an Fc-lacking 3E10 fragment to penetrate cells), which involves the presence of the nucleoside transporter ENT2 (see Weisbart et al., Sci. Rep. 5:12022. doi:10.1038 / srep12022. (2015); Zack et al., J. Immunol. 157, 2082-2088 (1996); Hansen et al., J. Biol. Chem. 282, 20790-20793 (2007)). Thus, in embodiments, antibodies utilized in the compositions, conjugates, and methods penetrate cells via an Fc-independent mechanism, which involves the presence of the nucleoside transporter ENT2.

[0316] Mutations in 3E10 that interfere with its ability to bind to DNA can render the antibody unable to penetrate the nucleus. Thus, the disclosed variants and humanized forms of the antibody generally maintain the ability to bind to nucleic acids, particularly DNA. In addition, 3E10 scFv has previously been shown to be able to penetrate living cells and the nucleus, with the efficiency of uptake being reduced in ENT2-deficient cells (Hansen, et al., J. Biol. Chem. 282, 20790-20793 (2007)). Thus, in embodiments, the disclosed variants and humanized forms of the antibody maintain the ability to penetrate the cell nucleus in an ENT2-dependent manner.

[0317] B. Therapeutic Agents As used herein, the terms "therapeutic agent," "payload moiety," "payload," "drug moiety," "drug," "antineoplastic agent," "bioactive molecule," "active molecule," and "chemotherapeutic warhead" all refer to a molecule that has a biological, cytotoxic, or therapeutic effect on a cell. In embodiments, the drug or active molecule can be an inorganic molecule, an organic molecule, a small organic molecule, a drug compound, a peptide, or a polypeptide. In embodiments, the drug or active molecule can be a functional nucleic acid such as an oligonucleotide or a polynucleotide.

[0318] 1.Low molecules In embodiments, the therapeutic agent is an anti-tumor drug or active molecule. In embodiments, the therapeutic agent or active molecule is a cytotoxic agent. The term "cytotoxic agent" generally refers to a substance that destroys cells. In embodiments, the anti-tumor drug or active molecule can be a DNA damage inducer, a DNA repair inhibitor, an immunomodulatory molecule, an alkylating agent, a microtubule inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, an adoptive cell therapy, or a topoisomerase inhibitor, or any other cytotoxic agent or chemotherapeutic small molecule known in the art.

[0319] In the ADC embodiments provided herein, P is a DNA damage inducer, a DNA repair inhibitor, an immunomodulatory molecule, an alkylating agent, a microtubule inhibitor, an immune checkpoint inhibitor, an angiogenesis inhibitor, an adoptive cell therapy, or a topoisomerase inhibitor.

[0320] In embodiments, the anti-tumor drug or active molecule is maytansinoid, benzodiazepine, auristatin, tecan, taxoid, CC-1065, (4S)-4,11-diethyl-4,9-dihydroxy-1,4-dihydro-3H,14H-pyrano[3',4'.6,7]indolizino[1,2-b]quinoline-3,14-dione (SN38), exatecan, monomethylauristatin E (MMAE), monomethylauristatin F (MMAF), pyrrolobenzodiazepine (PBD), PROteolysis Targeting Agent (PTAG). The antitumor drug may be selected from Chimera (PROTAC), delextecan (Dxd), calicheamicin, duocarmycin, stimulator of interferon genes (STING) agonists, PNU-159682, NMS249, IMGN camp 1, duocarmycin hydroxybenzamide azaindole (DUBA), and their prodrugs. In embodiments, the antitumor drug may be Pseudomonas aeruginosa exotoxin PE38, calicheamicin, diphtheria toxin, irinotecan, duocarmycin, exatecan, Staphylococcus aureus enterotoxin A / E-120, doxorubicin, tubulidine, antibacterial antibiotic, ciguatoxin, lysine, or urease. In embodiments, the antitumor drug may be N(2')-deacetyl-N(2')-(3-mercapto-1-oxopropyl)-maytansine ("DM1"). In embodiments, the antineoplastic agent can be N2'-deacetyl-N2'-(4-mercapto-4-methyl-1-oxopropyl)-maytansine ("DM4"). In embodiments, the antineoplastic agent can be SN38. In embodiments, the antineoplastic agent can be PNU-159682, NMS249. In embodiments, the antineoplastic agent can be NMS249.

[0321] In embodiments, the anti-tumor drug or active molecule is doxorubicin (ADRIAMYCIN®), cisplatin, carboplatin, bleomycin sulfate, carmustine, chlorambucil (LEUKERAN®), cyclophosphamide (CYTOXAN®; NEOSAR®), lenalidomide (REVLIMID®), bortezomib (VELCADE®), dexamethasone, mitoxantrone, etoposide, cytarabine, bendamustine (TREANDA®), rituximab (RITUXAN®), ifo The agent can be sufamide, vincristine (ONCOVIN®), fludarabine (FLUDARA®), thalidomide (THALOMID®), alemtuzumab (CAMPATH®), ofatumumab (ARZERRA®), everolimus (AFINITOR®, ZORTRESS®), carfilzomib (KYPROLIST), a proteasome inhibitor, a thalidomide analog that is an immunomodulatory drug (IMiD), a Bet inhibitor, or any other cytotoxic or chemotherapeutic agent known in the art.

[0322] In some embodiments, the drug moiety is selected from any of the following: TIFF2026501556000051.tif192165TIFF2026501556000052.tif182165TIFF202 6501556000053.tif213165TIFF2026501556000054.tif226165TIFF20265015560 00055.tif197165TIFF2026501556000056.tif201165TIFF2026501556000057.t if164165TIFF2026501556000058.tif233165TIFF2026501556000059.tif155165

[0323] In embodiments of the ADCs provided herein, P is a maytansinoid. In embodiments, the P linker comprises -Phe-Gln- is N(2')-deacetyl-N(2')-(3-mercapto-1-oxopropyl)-maytansine ("DM1"). In embodiments, P is N2'-deacetyl-N2'-(4-mercapto-4-methyl-1-oxopropyl)-maytansine ("DM4").

[0324] In embodiments of the ADCs provided herein, P is (4S)-4,11-diethyl-4,9-dihydroxy-1,4-dihydro-3H,14H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14-dione (SN38).

[0325] In the ADC embodiments provided herein, A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is SMCC, and P is DM1.

[0326] In embodiments of the ADCs provided herein, A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is O-succinyl, and P is SN38. Thus, in embodiments, L is a cleavable linker.

[0327] In the ADC embodiments provided herein, A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is CL2A, and P is SN38.

[0328] In the ADC embodiments provided herein, A is the 3E10 antibody or an antigen-binding fragment thereof or a variant thereof, L is MC-vc-PAB, and P is PNU-159682.

[0329] In the ADC embodiments provided herein, A is the 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is MC-vc-PAB, and P is NMS249.

[0330] In embodiments, the disclosure provides antibody-drug conjugates (ADCs) having the formula A-(L-Pr)q, where A is selected from the group consisting of (a) a light chain variable region (VL) complementarity determining region (CDR) 1 comprising the amino acid sequence of RASKSVSTSSYSYMH (SEQ ID NO: 9), (b) a VL CDR2 comprising the amino acid sequence of YASYLES (SEQ ID NO: 10), (c) a VL CDR3 comprising the amino acid sequence of QHSREFPWT (SEQ ID NO: 11), (d) a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of NYGMH (SEQ ID NO: 15), (e) a VH CDR2 comprising the amino acid sequence of YISSGSSTIYYADTVKG (SEQ ID NO: 4), and (f) a VH CDR3 comprising the amino acid sequence of RGLLLDY (SEQ ID NO: 5). and CDR3, wherein L is O-succinyl; P is SN38; r is an integer of 1 to 4; and q is an integer of 1 to 16.

[0331] In embodiments, the ADCs disclosed herein have the formula A-(L-Pr)q, where A is a 3E10 antibody or antigen-binding fragment thereof or variant thereof, L is a linker, P is a payload described herein, r is an integer between 1 and 4, and q is an integer between 1 and 16; wherein the 3E10 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of XYGMH (SEQ ID NO: 16), where X=D, N, R, L, or K, corresponding to amino acid residue 31 of the 3E10 heavy chain, and D31 is wild-type 3E10 CDR1.

[0332] C. Linker In embodiments, the universal antibody-drug conjugates (ADCs) provided herein comprise a linker. The linkers (L) described herein can be used to link or conjugate the 3E10 antibody or antigen-binding fragment or variant thereof to biologically active molecules, such as oligonucleotides, drugs, antitumor agents, or toxic agents, chemotherapeutic agents, or warheads used to kill cancer cells. As used herein, the term "linker" includes, but is not limited to, any known linker for use in antibody-drug conjugates known in the art. In embodiments, the ADC conjugates comprise one, two, three, four, or more linkers.

[0333] 1. Site and method of conjugation In embodiments, the one or more amino acids suitable for conjugation of the 3E10 antibody or cell-penetrating fragment thereof provided herein are selected from lysine, cysteine, histidine, arginine, aspartic acid, glutamine, serine, threonine, and tyrosine. In embodiments, the one or more amino acids suitable for conjugation are introduced by substitution of one or more amino acids in the 3E10 antibody or cell-penetrating fragment thereof. In embodiments, the one or more conjugated amino acids are lysine or arginine, and conjugation is via amine conjugation. In embodiments, the one or more conjugated amino acids are glutamine (Gln), and conjugation is via transglutaminase (TGase)-mediated enzymatic conjugation. In some embodiments, the one or more conjugated amino acids are cysteine ​​(Cys), and conjugation is via thiol conjugation. For example, primary amines in the 3E10 antibody or cell-penetrating fragment thereof provided herein can be modified using an NHS-PEG reagent.

[0334] In embodiments, the method of site-specific conjugation is transglutaminase-mediated. Transglutaminases (TGases), which also include bacterial transglutaminase (BTG), are a family of enzymes that catalyze the formation of a covalent bond between the γ-carbonyl-amide group of glutamine and the primary amine group of lysine.

[0335] A peptide or antibody can be a substrate for transglutaminase according to the methods of the present disclosure. Thus, in some embodiments, the peptide or antibody contains a Gln or Lys residue, particularly a Gln residue. In some embodiments, the peptide or antibody is not a substrate for transglutaminase, but one or more Gln or Lys residues, particularly Gln residues, are inserted into the peptide or antibody sequence to make the peptide or antibody a substrate for transglutaminase. In embodiments, the Gln or Lys residue can be inserted at any position within the peptide or antibody sequence, but is preferably inserted at a position where the physiological properties of the peptide, such as therapeutic activity, are not affected to the extent that the peptide is no longer useful, for example, in therapeutic intervention. Insertion of amino acid residues into a peptide can be achieved by standard techniques known to those skilled in the art, such as post-translational chemical modification or transgenic technology, as described in U.S. Pat. Nos. 11,123,439 and US / 2016 / 0355859, the contents of which are incorporated herein by reference.

[0336] Such transglutaminases accept substrates other than lysine as amine donors and are therefore used to modify proteins, including antibodies, with suitable acceptor glutamines. (Josten et al., J. Immunol. Methods, 240, 47-54 (2000); Mindt et al., Bioconjugate Chem. 19, 271-278 (2008); Dennler et al., in Antibody Drug Conjugates (Ducry, L., Ed.), pp. 205-215, Humana Press. (2013), the contents of which are incorporated herein by reference.) Transglutaminases have been used to conjugate drugs to antibodies containing artificial glutamine tags, which are acceptor glutamine residues introduced into antibodies by genetic engineering (Strop et al., Chem. Biol. 20, 161-167 (2013)). Furthermore, transglutaminases can be ...Kabat et al., J. Immunol. Methods, 240, 47-54 (2000)). While N297 (Kabat EU numbering) is the only γ-carbonyl amide donor and therefore acceptor glutamine for bacterial transglutaminase in the backbone of an unglycosylated IgG1 molecule, if the antibody is glycosylated at position N297 (Kabat EU numbering) of the heavy chain, no acceptor glutamine is present in the backbone of the IgG1. In summary, bacterial transglutaminase can be used for conjugation of an amine donor substrate, e.g., a drug-linker construct, at the acceptor glutamine residue of an antibody. Such an acceptor glutamine can be introduced by mutational engineering of the antibody or by generating an unglycosylated antibody. Such an unglycosylated antibody can be introduced by deglycosylation using N-glycosidase F (PNGase F) or by mutating N297 of the glycosylation site (Kabat EU numbering) of the heavy chain to any amino acid other than N.Enzymatic conjugation of such non-glycosylated antibodies using bacterial transglutaminase has been described for non-glycosylated antibody variants containing the N297D, N297Q, or N297S mutations (see U.S. Pat. Nos. 9,764,038 and 9,764,038, the contents of which are incorporated by reference). Enzymatic conjugation of such non-glycosylated antibodies with transglutaminase generally results in an AOC with a DAR of 2, in which both heavy chains are specifically functionalized at position Q295 (Kabat EU numbering). Only the heavy chain mutation N297Q provides an additional conjugation site per heavy chain. Conjugation of such variants results in an AOC with a DAR of 4, in which both heavy chains are specifically functionalized at positions Q295 and Q297.

[0337] In embodiments, the chemical modification strategy utilized to create the ADCs described herein is lysine conjugation. Lysine residues in proteins, such as antibodies, possess primary amine groups (-NH) in their side chains, making them suitable targets for chemical modification. These primary amine groups can react with a variety of chemical reagents, including small molecules or polymers (e.g., cleavable and non-cleavable linkers).

[0338] In embodiments, lysine conjugation can be either site-specific or random. Site-specific conjugation allows for the targeting of specific lysine residues within a protein, such as an antibody, ensuring precise control of the modification. In contrast, random conjugation involves the modification of lysine residues without selectivity.

[0339] However, as described in Example 26 and Figures 39A-39C, tryptic peptide mapping of 3E10 conjugated at lysine residues revealed that both VL and VH V region lysines were observed to be modified, with VL K53 being the most commonly modified lysine, occurring in approximately two-thirds of cases. Furthermore, humanization studies described in WO2023 / 168352 demonstrated that any modification to VL K53 adversely affected the immunoreactivity of the humanized 3E10-D31N monoclonal antibody (V66) to nucleic acids. This is further demonstrated in Example 27, which demonstrates greater uptake of 3E10-D31N monoclonal antibody (V66) oligonucleotide conjugates in A427 cells using transglutaminase-mediated enzymatic conjugation, and in Example 28, which utilizes a hidden disulfide linker (SPDMV) conjugate (transglutaminase-SPDMV) and provides the highest exon skipping (approximately 15%), representing a more than 10-fold improvement over lysine-PEG8 AOC.

[0340] In embodiments, the conjugation strategy is selected from one of the following: TIFF2026501556000060.tif103165

[0341] Lysine-linked In some embodiments of the ADCs described herein, a linker moiety is conjugated to an antibody or antigen-binding fragment thereof via an amine bond at one or more surface-exposed lysine residues on the antibody or antigen-binding fragment. Generally, lysine conjugation is a random process with respect to which lysine is conjugated to the linker-payload. However, the primary, secondary, tertiary, and / or quaternary structural context surrounding a particular lysine residue may result in some preference for conjugation at a particular lysine. Many different chemistries for linking payloads to proteins at lysine groups are known in the art. For example, activated esters on drug-linker conjugates, often O-succinimide reagents such as N-hydroxysuccinimidyl (NHS) or sulfo-NHS esters, can react with lysine residues on antibodies to achieve conjugation via an amide bond, or stable amidine bonds can be generated on antibodies by reaction of imidoester compounds, such as Traut's reagent, with antibody lysine residues. Further description of lysine conjugation techniques can be found, for example, in Walker, JM, et al., "Antibody-drug conjugates," Humana Press (2013); Bhat, AS, et al., "The next step in homogenous bioconjugate development: optimizing payload placement and conjugate composition," BioProcess International (2014); and Jain, N., et al., "Current ADC linker chemistry," Pharm. Res., 32:3526-40 (2015), the disclosures of which are incorporated herein by reference in their entireties for all purposes.

[0342] Thus, in some embodiments, the present disclosure provides a compound of formula A-(LP r ) qwherein A is the 3E10 antibody or antigen-binding fragment thereof described herein; L is a linker described herein; P is a payload moiety described herein; r is an integer between 1 and 4; and q is an integer between 1 and 16; and wherein L is conjugated to A via a lysine moiety.

[0343] Advantageously, because the structural regions of antibodies contain many lysine residues, lysine conjugation can be used to generate ADC molecules with high drug-to-antibody ratios (DARs). Thus, in some embodiments, compositions comprising ADCs in which a drug is conjugated to an antibody or antigen-binding fragment thereof described herein using lysine linkages will have an average DAR of at least 4 (average of at least four drug moieties linked to each antibody). In some embodiments, such compositions have an average DAR of at least 6. In some embodiments, such compositions have an average DAR of at least 8. In some embodiments, such compositions have an average DAR of at least 10. In some embodiments, such compositions have an average DAR of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or more.

[0344] Thus, in some embodiments, the compositions include ADCs in which a drug is conjugated to an antibody or antigen-binding fragment thereof described herein using a lysine linkage, where r is 1 and q is at least 4. In some embodiments, r is 1 and q is at least 6. In some embodiments, r is 1 and q is at least 8. In some embodiments, r is 1 and q is at least 10. In some embodiments, r is 1 and q is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or more. In some embodiments, the compositions include ADCs in which a drug is conjugated to an antibody or antigen-binding fragment thereof described herein using a lysine linkage, where r is 2 (e.g., the linker is a branched linker), and q is at least 2. In some embodiments, r is 2 and q is at least 3. In some embodiments, r is 2 and q is at least 4. In some embodiments, r is 2 and q is at least 5. In some embodiments, r is 2 and q is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, or more. In still other embodiments, the compositions include ADCs in which a drug is conjugated to an antibody or antigen-binding fragment thereof described herein using a lysine linkage, and the linker is highly branched, e.g., r is at least 3. In some embodiments, r is at least 4. In some embodiments, r is 1, at least 5, 6, 7, 8, or more.

[0345] b. cysteine-linked In some embodiments of the ADCs described herein, a linker moiety is conjugated to an antibody or antigen-binding fragment thereof via a sulfide bond at one or more surface-exposed cysteine ​​residues on the antibody or antigen-binding fragment thereof. In some embodiments of the ADCs described herein, a linker moiety is conjugated to an antibody or antigen-binding fragment thereof via the thiol side chain of one or more cysteine ​​residues on the antibody or antigen-binding fragment thereof. Generally, antibodies have no free thiols, and all cysteine ​​residues form disulfide bonds. In human IgG1, which is commonly used in current ADCs, there are four interchain disulfide bonds and 12 intrachain disulfide bonds. The four interchain disulfides, which are generally not important for the structural stability of IgG1, can be selectively reduced under mild conditions to yield two, four, six, or eight free thiols, while leaving the 12 intrachain disulfides intact. Due to the limited number of conjugation sites and the distinct reactivities of the thiol groups, cysteine-based conjugation allows for controlled DAR and heterogeneity. Engineered Cys residues can also be used for site-specific conjugation without the need for partial reduction of endogenous disulfide bonds, for example, using EnCys-mAb technology. Many different chemistries for linking payloads to proteins at cysteine ​​residues are known in the art. For example, interchain disulfide bonds can be reduced using a reducing agent to liberate eight nucleophilic cysteine ​​residues that can then be conjugated with drug-linker conjugates. This approach generates ADCs with heterogeneous conjugation sites and different numbers of drugs, resulting in drug-to-antibody ratios (DARs) ranging from 0 to 8. Alternatively, partial reduction with either dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP) can be used to disrupt the inter-heavy chain disulfide and liberate a free Cys for drug conjugation, while treatment with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) results in a drug conjugate attached to the Cys residue normally involved in the inter-heavy chain disulfide. Further description of cysteine ​​conjugation techniques can be found, for example, in

[0346] Behrens, C.R. et al., "Methods for site-specific drug conjugation to antibodies," mAbs. 2014, 6(1):46-53; Dennler, P. et al., "Antibody conjugates: from heterogeneous populations to defined reagents," Antibodies. 2015, 4:197-224; and Agarwal, P. et al., "Site-specific antibody-drug conjugates: the nexus of bioorthogonal chemistry, protein engineering, and drug development," Bioconjugate Chem. 2015, 26:176-192, the disclosures of which are incorporated herein by reference in their entireties for all purposes.

[0347] Thus, in some embodiments, the present disclosure provides a compound of formula A-(LP r ) q wherein A is the 3E10 antibody or antigen-binding fragment thereof as described herein; L is a linker as described herein; P is a payload moiety as described herein; r is an integer between 1 and 4; and q is an integer between 1 and 16, wherein L is conjugated to A via a cysteine ​​moiety.

[0348] Advantageously, due to the limited number of conjugation sites and well-defined reactivity of thiol groups, cysteine-based conjugation can be used to generate ADC molecules with controlled DAR and heterogeneity.

[0349] Thus, in some embodiments, compositions comprising ADCs used to conjugate drugs to antibodies or antigen-binding fragments thereof described herein using cysteine ​​linkages have an average DAR of at least 2. In some embodiments, such compositions have an average DAR of at least 3. In some embodiments, such compositions have an average DAR of at least 4. In some embodiments, such compositions have an average DAR of at least 5. In some embodiments, such compositions have an average DAR of at least 6. In some embodiments, such compositions have an average DAR of at least 7. In some embodiments, such compositions have an average DAR of about 8. In some embodiments, such compositions have an average DAR of at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, or about 8.

[0350] Thus, in some embodiments, compositions comprising ADCs that conjugate a drug to an antibody, or antigen-binding fragment thereof, described herein using a cysteine ​​linkage, r is 1 and q is at least 2. In some embodiments, r is 1 and q is at least 3. In some embodiments, r is 1 and q is at least 4. In some embodiments, r is 1 and q is at least 5. In some embodiments, r is 1 and q is at least 6. In some embodiments, r is 1 and q is at least 7. In some embodiments, r is 1 and q is about 8. In some embodiments, r is 1 and q is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, or 8. In some embodiments, compositions comprising ADCs that conjugate a drug to an antibody, or antigen-binding fragment thereof, using a cysteine ​​linkage, r is 2 (e.g., the linker is a branched linker) and q is at least 2. In some embodiments, r is 2 and q is at least 3. In some embodiments, r is 2 and q is at least 4. In some embodiments, r is 2 and q is at least 5. In some embodiments, r is 2 and q is at least 6. In some embodiments, r is 2 and q is at least 7. In some embodiments, r is 2 and q is about 8. In some embodiments, r is 2 and q is at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, or about 8. In still other embodiments, compositions comprising ADCs that conjugate drugs to antibodies or antigen-binding fragments thereof described herein using cysteine ​​linkages, the linker is highly branched, e.g., r is at least 3. In some embodiments, r is at least 4. In some embodiments, r is at least 5, 6, 7, 8, or more.

[0351] C. Transglutaminase-based ligation In some embodiments of the ADCs described herein, the linker moiety is conjugated to the antibody or antigen-binding fragment thereof via the primary amide side chains of one or more glutamine residues on the antibody or antigen-binding fragment thereof. Generally, transglutaminase from Streptomyces mobaraensis catalyzes transpeptidation, in which a primary amine-containing linker is covalently attached to the primary amide side chain of a specific glutamine (Q295) within a deglycosylated antibody, resulting in an ADC containing a defined DAR of 2 (one conjugation site per heavy chain) (Jeger et al., "Site-specific and stoichiometric modification of antibodies by bacterial transglutaminase," Angew. Chem. Int. Ed. Engl. 2010 49:9995-9997; and Dennler et al., "Transglutaminase-based chemo-enzymatic conjugation approach yields homogeneous antibody-drug conjugates," Bioconjugate Chem, 2014, 25:569-578, the disclosures of which are incorporated herein by reference in their entirety for all purposes). This N297Q mutation prior to conjugation provides two additional reactive sites (DAR = 4). This method is highly advantageous for producing practical ADCs because glycosidases and transglutaminases directly modify and conjugate natural mAbs to payloads without the need for genetic engineering. Another version using peptide sequence-specific transglutaminases can also be used (Strop et al., “Location matters: site of conjugation modulates stability and pharmacokinetics of antibody drug conjugates,” Chem. Biol. 2013, 20:161-167). This enzyme recognizes and exploits the genetically engineered LLQG motif to generate site-specific antibody-drug conjugation.

[0352] Thus, in some embodiments, the present disclosure provides a compound of formula A-(LP r ) q wherein A is the 3E10 antibody or antigen-binding fragment thereof as described herein, L is a linker as described herein, P is a payload moiety as described herein, r is an integer between 1 and 4, and q is an integer between 1 and 16, and wherein L is conjugated to A via a glutamine moiety.

[0353] Thus, in some embodiments, compositions comprising ADCs that conjugate drugs to antibodies or antigen-binding fragments thereof described herein using glutamine linkages have an average drug-antibody ratio (DAR) of about 2. In some embodiments, such compositions have an average DAR of at least 2. In some embodiments, such compositions have an average DAR of at least 3. In some embodiments, such compositions have an average DAR of about 4. In some embodiments, such compositions have an average DAR of about 2, at least 2, at least 3, or at least 4.

[0354] Thus, in some embodiments, compositions comprising ADCs that conjugate a drug to an antibody or antigen-binding fragment thereof described herein using glutamine linkages, r is 1 and q is about 2. In some embodiments, r is 1 and q is at least 2. In some embodiments, r is 1 and q is at least 3. In some embodiments, r is 1 and q is at least 4. In some embodiments, r is 1 and q is about 2, at least 2, at least 3, or about 4. In some embodiments, compositions comprising ADCs that conjugate a drug to an antibody or antigen-binding fragment thereof described herein using glutamine linkages, r is 2 (e.g., the linker is a branched linker) and q is about 2. In some embodiments, r is 2 and q is at least 2. In some embodiments, r is 2 and q is at least 3. In some embodiments, r is 2 and q is about 4. In some embodiments, r is 2 and q is about 2, at least 2, at least 3, or about 4. In still other embodiments, compositions comprising ADCs that conjugate drugs to antibodies or antigen-binding fragments thereof described herein using glutamine linkages, the linker is highly branched, e.g., r is at least 3. In some embodiments, r is at least 4. In some embodiments, r is at least 5, 6, 7, 8, or more.

[0355] 2. Cleavable Linker In embodiments, the linker comprises a cleavable linker. As used herein, "cleavable linker" refers to a linker that connects two or more molecules and can then be cleaved when exposed to a drug. A cleavable linker can include a chemically or enzymatically unstable or degradable bond. A cleavable linker generally relies on an intracellular process to release the drug, such as reduction in the cytoplasm, exposure to acidic conditions in lysosomes, or cleavage by specific proteases or other enzymes within the cell. A cleavable linker generally incorporates one or more chemical bonds that are either chemically or enzymatically cleavable, while the remainder of the linker is not cleavable. In embodiments, the cleavable linker is an acid-labile linker (Linker No. 215), a phosphatase linker (Linker No. 216), a glucuronidase linker (Linker No. 217), a cathepsin-B cleavable linker (Linker No. 218), a cathepsin-L cleavable linker (Linker No. 339), a protease-sensitive linker (Linker No. 219), a photolabile linker (Linker No. 220), or a disulfide (SPDMV) transglutaminase-containing linker (Link...

Claims

1. Formula (I): A-(L-P ｒ ) ｑ (Formula I) A complex of formula (I): A is an antibody, or an antigen-binding fragment thereof, comprising a heavy chain variable region (VH) CDR1 comprising the amino acid sequence of SEQ ID NO: 58, a CDR2 comprising the amino acid sequence of SEQ ID NO: 59, a CDR3 comprising SEQ ID NO: 60, a light chain variable region (VL) CDR1 comprising the amino acid sequence of SEQ ID NO: 61, a CDR2 comprising the amino acid sequence of SEQ ID NO: 62, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 63; L is a linker; P is a payload moiety; r is an integer from 1 to 4; q is an integer from 1 to 16; The complex.

2. The linker L is an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted arylene, an optionally substituted cycloalkylene, an optionally substituted heteroalkylene, an optionally substituted heteroarylene, an optionally substituted heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -S(O)-, -S(O) ２ -, -OP(O)OR ａ -, -OP(O)OR ａ O-, -P(O)OR ａ O-, -O-, -CR ｂ ２ -, - [(CR ｂ ２ ) １－１２ O] １－５０ -, -C(O)-, -C(S)-, -C(=N-OH)-, -C(NR ａ ) -, -C(NH ２ Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -C(O)NR ａ SO ２ -, -SO ２ NR ａ C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -SC(O)NR ａ -, -NR ａ C(O)S-, -S(O) ｔ N (R ａ )-(wherein t is 1 or 2), -N(R ａ ) S (O) ｔ - (wherein t is 1 or 2), and -X ＡＡ -, - or -; Each R ａ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R ｂ independently at each occurrence, hydrogen, halides, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , —C(O)NR ａ ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted alkenyl, optionally substituted fluoroalkenyl, optionally substituted cycloalkenyl, optionally substituted cycloalkenylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Two independent R ｂ group or R ａ and R ｂ can be joined together to form an optionally substituted ring; and -X ＡＡ is an amino acid sequence containing 1 to 6 amino acid moieties, The composite of claim 1.

3. -X ＡＡ 3. The conjugate of claim 2, wherein each amino acid moiety is independently selected from alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), valine (Val), citrulline (Cit), and homocitrulline (HoCit).

4. The linker L is an optionally substituted C １ ~C １８ Alkylene, —C≡C—, —CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ Cycloalkylene, -[CH ２ O] １－１８ -, -[CH ２ CH ２ O] １－１８ -, -[CH ２ CH ２ CH ２ O] １－１８ -, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -CR ｂ ２ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -SC(O)NR ａ -, -NR ａ C(O)S- and -X ＡＡ The conjugate of any one of claims 1 to 3, comprising one or more groups selected from:

5. The linker L is an optionally substituted C １ ~C １６ Alkylene, —C≡C—, —CR ａ =CR ａ -, optionally substituted phenylene, optionally substituted C ３ ~C ６ Cycloalkylene, -[CH ２ CH ２ O] １－１６ -, -[CH ２ CH ２ CH ２ O] １－１６ -, optionally substituted 5- to 6-membered heteroarylene, optionally substituted 5- to 20-membered heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -CR ｂ ２ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O- and -X ＡＡ The conjugate of any one of claims 1 to 4, comprising one or more groups selected from:

6. The linker L is —[C(R ｂ ) ２ ] １－１６ -, -C≡C-, -CR ａ =CR ａ -, -[CH ２ CH ２ O] １－１６ -, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -X ＡＡ -, and X １ , X ２ and X ３ independently for each occurrence, NR ａ , N., C.R. ｂ , S and O; The composite according to any one of claims 1 to 5.

7. The linker L is represented by formula (L-1): In formula (L-1), L Ａ is a linking moiety through which A is covalently attached to L'; L' is a bond, or an optionally substituted alkylene, an optionally substituted alkenylene, an optionally substituted alkynylene, an optionally substituted arylene, an optionally substituted cycloalkylene, an optionally substituted heteroalkylene, an optionally substituted heteroarylene, an optionally substituted heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -S(O)-, -S(O) ２ -, -OP(O)OR ａ O-, -O-, -CR ｂ ２ -, - [(CR ｂ ２ ) １－１２ O] １－５０ -, -C(O)-, -C(S)-, -C(NR ａ ) -, -C(NH ２ Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -C(O)NR ａ SO ２ -, -SO ２ NR ａ C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -SC(O)NR ａ -, -NR ａ C(O)S-, -S(O) ｔ N (R ａ ) -, -N(R ａ ) S (O) ｔ - and -X ＡＡ -, ... L Ｐ is a linking moiety through which P is covalently attached to L'; The composite according to any one of claims 1 to 6.

8. The conjugate of any one of claims 1 to 7, wherein the linker L comprises at least one cleavable moiety.

9. 9. The conjugate of claim 8, wherein the cleavable moiety comprises an acid-labile moiety, a reducibly-labile moiety, or an enzymatically-labile moiety.

10. the cleavable portion is and During the ceremony, Each R ａ’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; The complex according to claim 8 or 9.

11. The conjugate of any one of claims 9 to 10, wherein the cleavable moiety comprises the reductively labile moiety -S-S-.

12. The linker L is represented by formula (L-10): In formula (L-10), L Ａ is a bond, -NR ａ’ -, and -S-; L １ is a bond or an optionally substituted C １ ~C １８ Alkylene, —C≡C—, —CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ Cycloalkylene, -[CH ２ O] １－１８ -, -[CH ２ CH ２ O] １－１８ -, -[CH ２ CH ２ CH ２ O] １－１８ -, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -CR ｂ ２ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -SC(O)NR ａ -, -NR ａ C(O)S- and -X ＡＡ -, - or -; L Ｃ is selected from an acid labile moiety, a reductively labile moiety, and an enzymatically labile moiety; L ２ is a bond or an optionally substituted C １ ~C １８ Alkylene, —C≡C—, —CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ Cycloalkylene, -[CH ２ O] １－１８ -, -[CH ２ CH ２ O] １－１８ -, -[CH ２ CH ２ CH ２ O] １－１８ -, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -CR ｂ ２ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -OC(O)S-, -SC(O)O-, -OC(O)NR ａ -, -NR ａ C(O)O-, -SC(O)NR ａ -, -NR ａ C(O)S- and -X ＡＡ -, - or -; L Ｐ is a bond, -NR ａ’ , —S—, and —O—; Each R ａ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R ａ’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , —C(O)NR ａ ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent R ｂ groups taken together form an optionally substituted cycloalkyl, an optionally substituted aryl, an optionally substituted heterocycloalkyl, or an optionally substituted heteroaryl; and -X ＡＡ - is an amino acid sequence containing 1 to 4 amino acid moieties, The composite according to any one of claims 1 to 8.

13. L Ｃ but, is selected from During the ceremony, Each R ａ’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; The composite of claim 11.

14. L Ｃ The complex of claim 12 or 13, wherein is -S-S-.

15. The linker L is represented by formula (L-11): In formula (L-11), L Ａ is selected from a bond, —NH—, and —S—; L １ is a bond or -[C(R ｂ ) ２ ] １－１６ -, -C≡C-, -CR ａ =CR ａ -, -[CH ２ CH ２ O] １－１６ -, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -X ＡＡ -, -OC(O)NR ａ -, -NR ａ C(O)O-, containing one or more groups selected from L ２ is a bond or -[C(R ｂ ) ２ ] １－１６ -, -C≡C-, -CR ａ =CR ａ -, -[CH ２ CH ２ O] １－１６ -, -NR ａ -, -N=CR ａ -, -CR ａ =N-, -S-, -OP(O)OR ａ O-, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, -OC(O)O-, -X ＡＡ -, -OC(O)NR ａ -, -NR ａ C(O)O-, containing one or more groups selected from L Ｐ is a bond, -NR ａ’ -O-; Each R １ are independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R １ the groups taken together form an optionally substituted cycloalkyl; Each R ２ are independently selected from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or both R ２ the groups taken together form an optionally substituted cycloalkyl; Each R ａ is independently at each occurrence hydrogen, optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ selected from cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, and optionally substituted 5- to 6-membered heteroaryl; Each R ａ’ is independently selected at each occurrence from hydrogen and optionally substituted alkyl; Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent R ｂ groups taken together form an optionally substituted cycloalkyl; and -X ＡＡ - is an amino acid sequence containing 2 to 4 amino acid moieties, The composite according to any one of claims 1 to 7.

16. The linker L is represented by formula (L-12): In formula (L-12), L Ａ is selected from a bond and —NH—; L １’ は、-[C(R ｂ ) ２ ] １－１０ -、-[CH ２ CH ２ O] １－１０ -、-NR ａ -、-O-、-C(O)-、-C(O)O-、-OC(O)-、-C(O)NR ａ -、-NR ａ C(O)-、-OC(O)O-、-X ＡＡ -、-OC(O)NR ａ -、-NR ａ C(O)O-、 containing one or more groups selected from L ２’ は、-[C(R ｂ ) ２ ] １－１０ -、-[CH ２ CH ２ O] １－１０ -、-NR ａ -、-O-、-C(O)-、-C(O)O-、-OC(O)-、-C(O)NR ａ -、-NR ａ C(O)-、-OC(O)O-、-X ＡＡ -、-OC(O)NR ａ -、-NR ａ C(O)O-、 containing one or more groups selected from L Ｐ represents a bond, and -NR ａ’ Selected from: Each R １ are independently hydrogen, optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both R １ The groups taken together form an optionally substituted C ３ ~C ６ Forming a cycloalkyl; Each R ２ are independently hydrogen, optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or both R ２ The groups taken together form an optionally substituted C ３ ~C ６ Forming a cycloalkyl; Each R ａ is independently at each occurrence hydrogen, optionally substituted C １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Fluoroalkyl and optionally substituted C ３ ~C ６ cycloalkyl; Each R ａ’ represents independently at each occurrence hydrogen and optionally substituted C １ ~C ６ alkyl; Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted C １ ~C ６ Alkyl, optionally substituted C １ ~C ６ Fluoroalkyl and optionally substituted C ３ ~C ６ cycloalkyl; or two independent R ｂ The groups taken together form an optionally substituted C ３ ~C ６ forming a cycloalkyl; and -X ＡＡ - is an amino acid sequence containing two or three amino acid moieties, The composite according to any one of claims 1 to 7.

17. At least one R １ or R ２ 17. The complex of claim 15 or 16, wherein is other than hydrogen.

18. At least one R １ is optionally substituted C １ ~C ８ The conjugate of any one of claims 15 to 17, wherein the alkyl is alkyl.

19. Each R １ are independently optionally substituted C １ ~C ８ The conjugate of any one of claims 15 to 17, wherein the alkyl is alkyl.

20. At least one R ２ is optionally substituted C １ ~C ８ The conjugate of any one of claims 15 to 19, wherein the alkyl is alkyl.

21. Each R ２ are independently optionally substituted C １ ~C ８ The conjugate of any one of claims 15 to 19, wherein the alkyl is alkyl.

22. The linker L is The complex according to any one of claims 1 to 10, wherein the complex is selected from

23. The linker L is The complex according to any one of claims 1 to 10,

24. The linker L is The complex according to any one of claims 1 to 10,

25. The linker L is The complex according to any one of claims 1 to 10,

26. The linker L is The complex according to any one of claims 1 to 16, selected from:

27. The conjugate of any one of claims 8 to 26, wherein the linker is a cleavable linker.

28. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a lysine of the 3E10 antibody or antigen-binding fragment thereof.

29. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a cysteine ​​of the antibody or antigen-binding fragment thereof.

30. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a histidine of the antibody or antigen-binding fragment thereof.

31. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to an arginine of the antibody or antigen-binding fragment thereof.

32. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to an aspartic acid of the antibody or antigen-binding fragment thereof.

33. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a glutamine of the antibody or antigen-binding fragment thereof.

34. 34. The conjugate of claim 33, having a drug-to-antibody ratio (DAR) of at least 4:

1.

35. 35. The conjugate of claim 33 or 34, wherein the linker is a branched linker linked to at least two copies of the oligonucleotide moiety.

36. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a serine of the antibody or antigen-binding fragment thereof.

37. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a threonine of the antibody or antigen-binding fragment thereof.

38. 28. The conjugate of claim 27, wherein the cleavable linker is conjugated to a tyrosine of the antibody or antigen-binding fragment thereof.

39. The linker L is represented by formula (L-20): In formula (L-20), L Ａ is a bond, -NR ａ’ -, and -S-; L ３ is a bond or -[C(R ｂ ) ２ ] １－８ -, -NR ａ -, -C(O)-, -C(S)-, -C(NR ａ ) -, -C(NH ２ Cl)-, -C≡C-, -CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ containing one or more groups selected from cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene; L Ｘ is an optionally substituted C １ ~C １８ Alkylene, —C≡C—, —CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ Cycloalkylene, -[CH ２ O] １－１８ -, -[CH ２ CH ２ O] １－１８ -, -[CH ２ CH ２ CH ２ O] １－１８ -, optionally substituted 5- to 18-membered heteroarylene, optionally substituted 3- to 20-membered heterocycloalkylene, -NR ａ -, -S-, -O-, -CR ｂ ２ -, -C(O)-, -C(S)-, -C(NR ａ ) -, -C(NH ２ Cl)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ - and -NR ａ C(O)—; L ４ is a bond or -[C(R ｂ ) ２ ] １－８ -, -NR ａ -, -C(O)-, -C(S)-, -C(NR ａ ) -, -C(NH ２ Cl)-, -C≡C-, -CR ａ =CR ａ -, optionally substituted 6- to 14-membered arylene, optionally substituted C ３ ~C ２０ containing one or more groups selected from cycloalkylene, optionally substituted 5- to 18-membered heteroarylene, and optionally substituted 3- to 20-membered heterocycloalkylene; L Ｐ is a bond, -NR ａ’ , —S—, and —O—; Each R ａ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; Each R ａ’ is independently selected at each occurrence from hydrogen, optionally substituted alkyl, and optionally substituted heteroalkyl; and Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , —C(O)NR ａ ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted alkyl, optionally substituted fluoroalkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkylalkyl, optionally substituted heteroaryl, and optionally substituted heteroarylalkyl; or two independent R ｂ the groups taken together form an optionally substituted cycloalkyl, an optionally substituted aryl, an optionally substituted heterocycloalkyl or an optionally substituted heteroaryl; The composite according to any one of claims 1 to 7.

40. The linker L is represented by formula (L-21): In formula (L-21), L Ａ is selected from a bond and —NH—; L Ｘ is an optionally substituted —[C(R ｂ ) ２ ] １－１６ -, -C≡C-, -CR ａ =CR ａ -, -[CH ２ CH ２ CH ２ O] １－１６ -, -NR ａ -, -O-, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, containing one or more groups selected from L Ｐ represents a bond, and -NR ａ’ Selected from: Each R ａ is independently at each occurrence hydrogen, optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ selected from cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, and optionally substituted 5- to 6-membered heteroaryl; Each R ａ’ represents independently at each occurrence hydrogen and optionally substituted C １ ~C ６ alkyl; and Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent R ｂ the groups taken together form an optionally substituted cycloalkyl; The composite according to any one of claims 1 to 7.

41. The linker L is of formula (L-22a) or (L-22b): In formulas (L-22a) and (L-22b), L Ａ is selected from a bond and —NH—; L Ｘ is an optionally substituted —[C(R ｂ ) ２ ] １－１０ -, -C≡C-, -CR ａ =CR ａ -, -[CH ２ CH ２ CH ２ O] １－１０ -, -NR ａ -, -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -C(O)NR ａ -, -NR ａ C(O)-, containing one or more groups selected from L Ｐ represents a bond, and -NR ａ’ Selected from: Each R ａ is independently at each occurrence hydrogen, optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ selected from cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, and optionally substituted 5- to 6-membered heteroaryl; Each R ａ’ represents independently at each occurrence hydrogen and optionally substituted C １ ~C ６ alkyl; and Each R ｂ represents independently at each occurrence hydrogen, a halide, —OH, —SO ３ H, -OPO ３ H ２ , -PO ３ H ２ , -CO ２ R ａ , -NR ａ ２ , optionally substituted C １ ~C ８ Alkyl, optionally substituted C １ ~C ８ Fluoroalkyl, optionally substituted C ３ ~C ６ cycloalkyl, optionally substituted phenyl, optionally substituted benzyl, optionally substituted 5- to 10-membered heterocycloalkyl, optionally substituted 5- to 6-membered heteroaryl; or two independent R ｂ the groups taken together form an optionally substituted cycloalkyl; The composite according to any one of claims 1 to 7.

42. The linker L is The complex of claim 1, wherein the complex is selected from:

43. The conjugate of any one of claims 39 to 41, wherein the linker is a non-cleavable linker.

44. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a lysine of the antibody or antigen-binding fragment thereof.

45. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a cysteine ​​of the antibody or antigen-binding fragment thereof.

46. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a histidine of the antibody or antigen-binding fragment thereof.

47. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to an arginine of the antibody or antigen-binding fragment thereof.

48. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to an aspartic acid of the antibody or antigen-binding fragment thereof.

49. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a glutamine of the antibody or antigen-binding fragment thereof.

50. 50. The conjugate of claim 49, having a drug-to-antibody ratio (DAR) of at least 4:

1.

51. 51. The conjugate of claim 49 or 50, wherein the linker is a branched linker linked to at least two copies of the oligonucleotide moiety.

52. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a serine of the antibody or antigen-binding fragment thereof.

53. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a threonine of the antibody or antigen-binding fragment thereof.

54. 44. The conjugate of claim 43, wherein the non-cleavable linker is conjugated to a tyrosine of the antibody or antigen-binding fragment thereof.

55. -X ＡＡ 17. The conjugate of any one of claims 2 to 16, wherein each amino acid moiety is independently selected from alanine (Ala), arginine (Arg), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), phenylalanine (Phe), tryptophan (Trp), tyrosine (Tyr), valine (Val), citrulline (Cit), and homocitrulline (HoCit).

56. -X ＡＡ 17. The conjugate of any one of claims 2 to 16, wherein each amino acid moiety is independently selected from alanine (Ala), glycine (Gly), lysine (Lys), phenylalanine (Phe), valine (Val), and citrulline (Cit).

57. The amino acid sequence -X ＡＡ The conjugate of any one of claims 2 to 15, wherein - is selected from -Val-Cit-, -Cit-Val-, -Val-Ala-, -Ala-Val-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, -Gly-Gly-Gly-, -Gly-Gly-Phe-Gly-, -Gly-Phe-Gly-Gly-, -Gly-Gly-Gly-Phe-, -Phe-Gly-Gly-Gly-, and -Gly-Gly-Gly-Gly-Gly-.

58. The amino acid sequence -X ＡＡ The conjugate of any one of claims 2 to 16, wherein - is selected from -Val-Cit-, -Cit-Val-, -Val-Ala-, -Ala-Val-, -Phe-Lys-, -Lys-Phe-, -Ala-Ala-, -Val-Val-, -Gly-Gly-, -Ala-Ala-Ala-, and -Gly-Gly-Gly-.

59. 59. The conjugate of any one of claims 1 to 58, wherein the payload moiety P is a drug moiety.

60. The drug moiety is:

60. The conjugate of claim 59, selected from any of:

61. 61. The conjugate of any one of claims 1 to 60, wherein the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 9, CDR2 comprises the amino acid sequence of SEQ ID NO: 10, and CDR3 comprises the amino acid sequence of SEQ ID NO: 11, and the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 15, CDR2 comprises the amino acid sequence of SEQ ID NO: 4, and CDR3 comprises the amino acid sequence of SEQ ID NO:

5.

62. 61. The conjugate of any one of claims 1 to 60, wherein the VL CDR1 comprises the amino acid sequence of SEQ ID NO:29, CDR2 comprises the amino acid sequence of SEQ ID NO:10, and CDR3 comprises SEQ ID NO:11, and the VH CDR1 comprises the amino acid sequence of SEQ ID NO:15, CDR2 comprises the amino acid sequence of SEQ ID NO:26, and CDR3 comprises the amino acid sequence of SEQ ID NO:

5.

63. The conjugate of any one of claims 1 to 62, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 21 and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:

14.

64. 63. The conjugate of any one of claims 1 to 62, wherein the antibody or antigen-binding fragment thereof comprises a full-length light chain (LC) comprising the amino acid sequence of SEQ ID NO: 20 and a full-length heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:

13.

65. The antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO:85), 3E10-VL-H2 (SEQ ID NO:86), 3E10-VL-H3 (SEQ ID NO:87), 3E10-VL-H4 (SEQ ID NO:88), 3E10-VL-H5 (SEQ ID NO:89), and 3E10-VL-H6 (SEQ ID NO:90). Ｌ ), and a heavy chain variable domain (VH) comprising an amino acid sequence at least about 95% identical to an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E10-VH-H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E10-VH-H5 (SEQ ID NO:68), 3E10-VH-H6 (SEQ ID NO:69), and 3E10-VH-H7 (SEQ ID NO:70). Ｈ 63. The complex of any one of claims 1 to 62, comprising:

66. The antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising an amino acid sequence selected from the group consisting of 3E10-VL-H1 (SEQ ID NO: 85), 3E10-VL-H2 (SEQ ID NO: 86), 3E10-VL-H3 (SEQ ID NO: 87), 3E10-VL-H4 (SEQ ID NO: 88), 3E10-VL-H5 (SEQ ID NO: 89), and 3E10-VL-H6 (SEQ ID NO: 90). Ｌ ), and a heavy chain variable domain (VH) comprising an amino acid sequence selected from the group consisting of 3E10-VH-H1 (SEQ ID NO:64), 3E10-VH-H2 (SEQ ID NO:65), 3E10-VH-H3 (SEQ ID NO:66), 3E10-VH-H4 (SEQ ID NO:67), 3E10-VH-H5 (SEQ ID NO:68), 3E10-VH-H6 (SEQ ID NO:69), and 3E10-VH-H7 (SEQ ID NO:70). Ｈ 63. The complex of any one of claims 1 to 62, comprising:

67. The antibody or antigen-binding fragment thereof is (a) VL1 (SEQ ID NO: 85) and VH1 (SEQ ID NO: 64), (b) VL1 (SEQ ID NO: 85) and VH2 (SEQ ID NO: 65), (c) VL1 (SEQ ID NO: 85) and VH3 (SEQ ID NO: 66), (d) VL1 (SEQ ID NO: 85) and VH4 (SEQ ID NO: 67), (e) VL2 (SEQ ID NO: 86) and VH1 (SEQ ID NO: 64), (f) VL2 (SEQ ID NO: 86) and VH2 (SEQ ID NO: 65), (g) VL2 (SEQ ID NO: 86) and VH3 (SEQ ID NO: 66), (h) VL2 (SEQ ID NO: 86) and VH4 (SEQ ID NO: 67), (i) VL3 (SEQ ID NO: 87) and VH1 (SEQ ID NO: 64), (j) VL3 (SEQ ID NO: 87) and VH2 (SEQ ID NO: 65), (k) VL3 (SEQ ID NO: 87) 63. The conjugate of any one of claims 1 to 62, comprising a VL / VH pair selected from the group consisting of: (i) VH3 (SEQ ID NO: 66), (l) VL3 (SEQ ID NO: 87) and VH4 (SEQ ID NO: 67), (m) VL4 (SEQ ID NO: 88) and VH1 (SEQ ID NO: 64), (n) VL4 (SEQ ID NO: 88) and VH2 (SEQ ID NO: 65), (o) VL4 (SEQ ID NO: 88) and VH3 (SEQ ID NO: 66), (p) VL4 (SEQ ID NO: 88) and VH4 (SEQ ID NO: 67), (q) VL5 (SEQ ID NO: 89) and VH5 (SEQ ID NO: 68), (r) VL5 (SEQ ID NO: 89) and VH6 (SEQ ID NO: 69), (s) VL6 (SEQ ID NO: 90) and VH5 (SEQ ID NO: 68), and (t) VL6 (SEQ ID NO: 90) and VH6 (SEQ ID NO: 69).

68. 61. The conjugate of any one of claims 1 to 60, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising 3E10-VL-H6 (SEQ ID NO: 90) and a heavy chain variable domain (VH) comprising 3E10-VH-H6 (SEQ ID NO: 69).

69. the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) comprising the amino acid sequence (DIQMTQSPSSLSASLGDRATITCRASKTVSTSSYSYMHWYQQKPGQPPKLLIKYASYLESGVPSRFSGSGSGTDFTLTISSLQPEDAATYYCQHSREFPWTFGGGTKVEIK) (SEQ ID NO: 126); and a heavy chain variable domain (VH) comprising (EVQLVESGGGLVQPGGSLRLSCAASGFTFFSNYGMHWVRQAPGKGLEWVSYISSGSSTIYYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARRGLLLDYWGQGTTVTVSS) (SEQ ID NO: 127).

70. 70. A method of treating a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a conjugate of any one of claims 1 to 69.

71. 70. A method of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a conjugate of any one of claims 1 to 69.

72. 72. The method of claim 71, wherein the cancer is carcinoma, sarcoma, blastoma, papilloma, or adenoma.

73. 72. The method of claim 71, wherein the cancer is a metastatic cancer.

74. 74. The method of any one of claims 71 to 73, wherein the cancer is selected from the group consisting of bladder cancer, blood cancer, brain cancer, breast cancer, bone cancer, cervical cancer, colorectal cancer, endocrine cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatobiliary cancer, leukemia, lung cancer, lymphoma, melanoma, myeloma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, thyroid cancer, and uterine cancer.

75. 75. The method of claim 74, wherein the cancer is a skin cancer selected from the group consisting of basal cell carcinoma, squamous cell carcinoma, and melanoma.

76. 75. The method of claim 74, wherein the cancer is melanoma.

77. 72. The method of claim 71, wherein the cancer is ovarian cancer.

78. 72. The method of claim 71, wherein the cancer is colon cancer.

79. 72. The method of claim 71, wherein the cancer is breast cancer.

80. 72. The method of claim 71, wherein the cancer is BRCA2- breast cancer.

81. 72. The method of claim 71, wherein the cancer is triple-negative breast cancer.

82. 72. The method of claim 71, wherein the cancer is lung cancer.

83. 72. The method of claim 71, wherein the cancer is non-small cell lung cancer (NSCLC).

84. 74. The method of any one of claims 71 to 73, wherein the cancer is a cancer of the central nervous system.

85. 72. The method of claim 71, wherein the cancer is a neuroepithelial brain or spinal cord tumor selected from the group consisting of medulloblastoma, astrocytic tumor, oligodendroglial tumor, oligoastrocytic tumor, ependymal tumor, choroid plexus tumor, neuronal or mixed neuronal-glial tumor, pineal region tumor, embryonal tumor, or neuroepithelial tumor not otherwise classified.

86. 85. The method of any one of claims 71 to 84, wherein the administration is by parenteral administration.

87. 86. The method of claim 85, wherein the parenteral administration is intramuscular, intravenous, or subcutaneous administration.

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