Antibody-drug conjugates

Abstract

The present disclosure provides, among other things, conjugates comprising a target binding moiety, a payload moiety, and a linker. In some embodiments, the present disclosure provides pharmaceutical compositions comprising the provided conjugates. In some embodiments, the present disclosure provides methods of using the provided conjugates and compositions thereof to prevent or treat various conditions, disorders, or diseases. In some embodiments, the present disclosure provides techniques for manufacturing the provided conjugates and compositions thereof.

Description

[0001] Cross-reference to related applications This application claims priority to U.S. Provisional Application No. 63 / 581,306, filed September 8, 2023; U.S. Provisional Application No. 63 / 591,078, filed June 17, 2023; and U.S. Provisional Application No. 63 / 550,023, filed February 5, 2024, the entire contents of each of which are incorporated herein by reference. Background Technology

[0002] Antibody-drug conjugates (ADCs) can provide an efficient means of delivering drugs to target sites in tissues or organisms. Antibody recognition of targets (such as tumors) minimizes exposure of non-target tissues to toxic chemotherapeutic agents and limits side effects associated with the toxicity of “free” drugs (i.e., those not bound to a carrier such as an antibody). Therefore, there is a need to develop new therapeutic modalities, such as ADCs, optimized to deliver drugs to target sites in tissues or organisms while minimizing side effects. Summary of the Invention

[0003] This disclosure particularly covers the understanding that conjugate design (e.g., ADC design) can affect its structure, properties, activity, etc. In some embodiments, this disclosure provides conjugates that can provide various benefits and advantages (e.g., improved properties, improved activity, etc.). For example, in some embodiments, the provided conjugates particularly have improved solubility, reduced aggregation, improved activity, etc. In some embodiments, the provided conjugates contain a moiety that can improve solubility and / or reduce aggregation (e.g., a carbohydrate moiety, a PEG moiety, etc.). In some embodiments, the provided conjugates contain -(CH2CH2O)n- as described herein. In some embodiments, n is 1-24. In some embodiments, -(CH2CH2O)n- is located in the bond chain from the target-binding moiety to the load moiety (compared to side chains outside the bond chain from the target-binding moiety to the load moiety).

[0004] In some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 It is or contains divisible units; and L CO It is a covalent bond or spacer part.

[0005] In some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 Is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; L CO It is a covalent bond or optionally substituted. In this case, the carbonyl group is bonded to the effective load portion; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0006] In some implementation schemes, R LS It is or contains a sugar portion, making L C It is cleavable, for example, by an enzyme. In some embodiments, R LS It may contain a glucuronide moiety. In some embodiments, L C It can be cleaved by β-glucuronidase. In some implementations, R LS It may contain a galactoside moiety. In some embodiments, L C It can be cleaved by β-galactosidase. In some implementations, R LS Yes or include Or its derivatives. In some embodiments, R LS yes In some implementations, R LS Yes or include Or its derivatives. In some embodiments, R LS yes In some implementations, L C1 It is as described in this article, L C1S In some implementations, L CO It is a covalent bond. In some implementations, L C It is as described in this article, L C1 In some implementations, L C It is as described in this article, L C1S In some implementations, L C1S The benzene ring -CH2- in it is unsubstituted.

[0007] In some implementations, n is 1-24.

[0008] In some embodiments, this disclosure provides a conjugate, wherein the conjugate is a compound of formula A-1 or a salt thereof: , A-1 in: Each R A It is a target-binding component independently; s is 1-20; Each L T Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L TThe 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX Independently covalent bonds, consisting of groups present on the unattached target-binding portion and containing L X’ The reaction between groups present on the agent produces partially or optionally substituted divalent straight-chain or branched C atoms. 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each of a and b is independently 1-10; Each L X’ Independently covalent or optionally substituted divalent or multivalent straight or branched C 1-30 Aliphatic, of which L X’ The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 Independently, it is a divisible unit, or each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S-or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; Each L CO Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L CO The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each R P It is a separate payload component; t is 1-20; Each -Cy- is independently a divalent 3- to 20-membered ring having 0-10 independently substituted heteroatoms selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0009] In some implementations, a is 1 and b is 1.

[0010] In some embodiments, this disclosure provides a conjugate (e.g., a conjugate of formula A-1), wherein the conjugate is a compound of formula A-2 or a salt thereof: , A-2 in: R A It is the target-binding part; Each L TIndependently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX Independently covalent bonds, consisting of groups present on the unattached target-binding moiety and containing -L X -(CH2CH2O)nL ZP -L C -R P The reaction between groups present on the agent produces partially or optionally substituted divalent straight-chain or branched C atoms. 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Or each L X Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 Independently, it is a divisible unit, or each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S-Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; Each L CO Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L CO The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each R P It is a separate payload component; t is 1-20; Each -Cy- is independently a divalent 3- to 20-membered ring having 0-10 independently substituted heteroatoms selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0011] In some implementations, each L C1 It is an independent, divisible unit. In some implementations, each L is... C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S-、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S - where each variable is independent as described herein. In some implementations, each L C1 Independently, it is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or L C1S In some implementations, each L C1 Independently -Val-Ala-, -Val-Cit-, or L C1S In some implementations, each L C1 Independently -Val-Ala-, -Val-Cit-, or L C1S , where R LS It may contain a glucuronide moiety. In some embodiments, each L C1 Independently -Val-Ala-, -Val-Cit-, or L C1S , where R LS yes In some implementations, each L C1 Independently, it is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or And each L CO Independently covalent bonds or optionally substituted The carbonyl group is bonded to the effective loading portion. In some embodiments, the occurrence of n at least once is 1-24. In some embodiments, each occurrence of n is independently 1-24. In some embodiments, each L C1 Independently, it is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or Each L CO Independently covalent bonds or optionally substituted The carbonyl group is bonded to the effective loading portion, and the occurrence of n at least once is independently 1-24. In some embodiments, each L C1 Independently, it is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or Each L CO Independently covalent bonds or optionally substituted The carbonyl group is bonded to the effective loading portion, and each occurrence of n is independently 1-24. In some embodiments, each L C1 Independently, it is -Val-Ala-. In some implementations, each L C1 Independently, it is -Val-Ala-, and each LCO Independently or arbitrarily replaced In some implementations, each L C1 Independently, it is -Val-Ala-, and each L CO yes In some implementations, each L C1 Independently, it is -Val-Cit-. In some implementations, each L C1 Independently, it is -Val-Cit-, and each L CO Independently or arbitrarily replaced In some implementations, each L C1 Independently, it is -Val-Cit-, and each L CO yes In some implementations, each L C1 Independently In some implementations, each L C1 Independently And each L CO It is a covalent bond. In some implementations, each L... C1 Independently In some implementations, each L C1 Independently And each L CO It is a covalent bond.

[0012] In some embodiments, the target-binding moiety binds to the peptide. For example, in some embodiments, the target-binding moiety binds to HHLA2 or a fragment thereof.

[0013] In some embodiments, the target binding portion is or comprises an antibody or an antigen-binding fragment thereof. For example, in some embodiments, the target binding portion is or comprises an anti-HHLA2 antibody or an antigen-binding fragment thereof.

[0014] Those skilled in the art will understand that various payload portions can be utilized according to this disclosure, such as those reported for antibody-drug conjugates. For example, in some embodiments, the payload portion is or comprises camptothecin or a derivative or analogue thereof. In some embodiments, the payload portion is or comprises... In some implementations, the payload is In some implementations, each payload is .

[0015] In some embodiments, this disclosure provides various adapters. For example, in some embodiments, the provided adapter comprises a PEG moiety. In some embodiments, the provided adapter comprises -(CH2CH2O)n-, where n is 1-24. In some embodiments, the provided adapter comprises a cleavable unit, such as GGFG, directly bonded to the payload moiety. In some embodiments, the cleavable unit may be cleaved by an enzyme (e.g., an enzyme present at the target site). In some embodiments, the cleavable unit is or comprises a polypeptide, such as GGFG, Val-Ala, Val-Cit, etc. In some embodiments, the polypeptide is bonded to the provided method via -C(O)- at its C-terminus. This disclosure particularly provides antibody-drug conjugates in which there is no portion (e.g., no spacer portion) between the cleavable unit and the payload moiety. In some embodiments, there is a portion (e.g., a spacer portion) between the cleavable unit and the payload moiety. In some embodiments, the adapter is or comprises -L ZP -L C - where each variable is independent as described herein. In some implementations, the connector is or contains -(CH2CH2O)nL ZP -L C - where each variable is independent as described herein. In some implementations, the connector is or contains -L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described in this paper.

[0016] In some embodiments, this disclosure provides pharmaceutical compositions comprising the provided conjugates and pharmaceutically acceptable carriers. The provided techniques (e.g., compounds, conjugates, agents, compositions, etc.) are particularly useful for the prevention or treatment of various conditions, symptoms, or diseases. In some embodiments, the provided compounds, conjugates, agents, etc., are provided, for example, as pharmaceutical compositions when used as therapeutic agents. The compounds, conjugates, agents, etc., can be in various forms, such as salt forms. For example, in some embodiments, they can be present in various pharmaceutically acceptable salt forms. In some embodiments, the compounds, conjugates, agents, etc., in the composition can be present in one or more forms, such as one or more pharmaceutically acceptable salt forms.

[0017] In some embodiments, this disclosure provides methods for preventing or treating various conditions, symptoms, or diseases. In some embodiments, this disclosure provides a method for preventing a condition, symptom, or disease, comprising administering or delivering an effective amount of the provided compound, conjugate, agent, or composition to a subject susceptible to it. In some embodiments, this disclosure provides a method for treating a condition, symptom, or disease, comprising administering or delivering an effective amount of the provided compound, conjugate, agent, or composition to a subject suffering from it. In some embodiments, the condition, symptom, or disease is cancer. In some embodiments, the subject has cancer or is at risk of developing cancer. In some embodiments, the subject has a solid tumor or a blood cancer. In some embodiments, a solid tumor is or includes one or more of the following: kidney cancer, bone cancer, skin cancer, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, lung cancer, ovarian cancer, liver cancer, bile duct cancer, or thyroid cancer. In some embodiments, a blood cancer includes leukemia or lymphoma. In some embodiments, leukemia includes acute lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic leukemia, or acute leukemia. In some embodiments, lymphoma includes Hodgkin lymphoma (HL), non-Hodgkin's lymphoma, lymphocytic lymphoma, or diffuse large B-cell lymphoma (DLBCL). In some embodiments, the condition, symptom, or disease is associated with abnormal expression of an agent that can bind to the target binding site (e.g., HHLA2). In some embodiments, the conjugate is administered parenterally. In some embodiments, parenteral administration is or includes subcutaneous, intravenous, intramuscular, or intrasternal injection or infusion. In some embodiments, the conjugate is administered in combination with another therapeutic agent.

[0018] In some embodiments, this disclosure provides techniques, such as methods, reagents, intermediates, etc., for preparing various compounds (e.g., the conjugates described herein).

[0019] Various features, objectives, and advantages of the provided technology will become apparent in the following detailed description. However, it should be understood that while the detailed description indicates embodiments of the provided technology, it is given by way of illustration only and not limitation. Various changes and modifications to the provided technology will become apparent to those skilled in the art based on this disclosure, including the detailed description. Attached Figure Description

[0020] Figure 1A , Figure 1B and Figure 1C The provided conjugate can bind to the target. Figure 1A, Figure 1B and Figure 1C Binding curves were plotted for some anti-HHLA2 antibody drug conjugates and their binding to HHLA2+A204 cells. Naked antibodies against each of Hu2C4-H3L3 (1A), 65872 (1B), and 887-1DE (1C), an exatecan conjugate with a DAR of approximately 4 (exatecan_DAR4), and an exatecan conjugate with a DAR of approximately 8 (exatecan_DAR8) were tested. Controls included human IgG1 isotype (IgG1 control) and an exatecan conjugate of human IgG1 isotype with a DAR of approximately 8 (IgG1-exatecan_DAR8).

[0021] Figure 2 The provided conjugates can be internalized. Figure 2 Examples of the internalization of anti-HHLA2 antibodies Hu2C4-H3L3, 65872, and 887-1DE induced in A204 cells are depicted.

[0022] Figure 3A and Figure 3B The provided conjugates can kill target cells. Figure 3A and Figure 3B Dose-response curves are plotted for example evaluation data from anti-HHLA2 antibody drug conjugates Hu2C4-H3L3, 65872, and 887-1DE. 3A: DAR approximately 4. 3B: DAR approximately 8. Data are also included for human IgG1 isotype ethatecan conjugates (IgG1-ethatecan_DAR8) and ethatecan itself, with a DAR of approximately 8.

[0023] Figure 4 The provided conjugate can bind to the target. Figure 4 Binding curves were plotted for some anti-HHLA2 antibody drug conjugates and HEK cells overexpressing B7H7. The Hu2C4-H3L3 conjugates for DAR 8 LP-A, LP-B, and LP-C, and the 65872 conjugate for DAR 8 LP-A, LP-B, and LP-C were plotted. Controls included the Hu2C4-H3L3-PEG8-GGFG-ethanotecan DAR8 and the GGFG-ethanotecan conjugate for human IgG1 isotype DAR 8. As demonstrated herein, the presented conjugates can bind to the target.

[0024] Figure 5 The provided conjugates can kill target cells. Figure 5Some dose-response curves from example evaluations of anti-HHLA2 antibody drug conjugates are depicted. The Hu2C4-H3L3 conjugates of DAR 8 (LP-A, LP-B, and LP-C) and the 65872 conjugate of DAR 8 (LP-A, LP-B, and LP-C) are depicted. Controls include the Hu2C4-H3L3-PEG8-GGFG-ethanotecan DAR8 and the GGFG-ethanotecan conjugate of human IgG1 isotype DAR 8. As demonstrated herein, the presented conjugates can effectively kill target cells.

[0025] definition To facilitate understanding of the provided technology, certain terms are defined below. Further definitions of the following and other terms are set forth throughout this specification. Publications cited herein (e.g., patents, patent application publications, non-patent publications, etc.), websites, and other references are each incorporated herein by reference in their entirety; in any event of conflict, this disclosure shall prevail.

[0026] Chemical definition The compounds disclosed herein include those generally described above, and are further illustrated by the classes, subclasses, and species disclosed herein. Unless otherwise stated, the following definitions shall apply as used herein. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. Furthermore, the general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999 and “March's Advanced Organic Chemistry”, 5th edition, edited by Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0027] Aliphatic: As used herein, "aliphatic" refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units (but not aromatics), or a substituted or unsubstituted monocyclic, bicyclic, or polycyclic hydrocarbon ring that is fully saturated or contains one or more unsaturated units (but not aromatics), or a combination thereof. In some embodiments, the aliphatic group contains 1-50 aliphatic carbon atoms. In some embodiments, the aliphatic group contains 1-20 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-10 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-9 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-8 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-7 aliphatic carbon atoms. In other embodiments, the aliphatic group contains 1-6 aliphatic carbon atoms. In yet another embodiment, the aliphatic group contains 1 to 5 aliphatic carbon atoms, and in still another embodiment, the aliphatic group contains 1, 2, 3, or 4 aliphatic carbon atoms. Some aliphatic groups include straight-chain or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0028] Alkyl: As used herein, the term "alkyl" has its common meaning in the art and refers to a saturated aliphatic group, such as a straight-chain alkyl, branched alkyl, cycloalkyl (alicyclic) group, alkyl-substituted cycloalkyl, cycloalkyl-substituted alkyl, etc. In some embodiments, the alkyl group has 1-100 carbon atoms. In some embodiments, the alkyl group has about 1-30 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 1-25, 1-20, 1-15, 1-10, 3-30, etc.). In some embodiments, the alkyl group has 1-20 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 1-15, 1-10, etc.). In some embodiments, the straight-chain or branched alkyl group has about 1-20 carbon atoms (e.g., for a straight chain of C1-C1) in its main chain. 20 For C2-C branches 20The alkyl group may have about 1-10 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, etc.). In some embodiments, the cycloalkyl ring has about 3-10 carbon atoms (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 3-8, 3-7, 3-6, 4-6, 5-6, 5-8, etc.) in its ring structure, wherein such ring is monocyclic, bicyclic, or polycyclic, and may alternatively have about 5, 6, or 7 carbon atoms in the ring structure. In some embodiments, the alkyl group may be a lower alkyl group, wherein the lower alkyl group has 1-4 carbon atoms (e.g., C1-C4 for straight-chain lower alkyl groups).

[0029] Aryl: As used herein, the term "aryl" used alone or as a part of a larger portion of terms such as "aralkyl", "aralkyloxy" or "aryloxyalkyl", means, unless otherwise specified, a monocyclic, bicyclic, or polycyclic system having a total of 5 to 30 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 5-25, 5-20, 5-15, 5-14, 5-10, 5-9, etc.) ring members, wherein at least one ring in the system is aromatic. In some embodiments, the aryl group is a monocyclic, bicyclic, or polycyclic system having a total of five to fourteen ring members (e.g., 5-10, 6-14, 6-10, 5, 6, 9, 10, 14, etc.), wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3 to 7 ring members. In some embodiments, each monocyclic unit is aromatic. In some embodiments, each aromatic ring is independently 6-membered. In some embodiments, each aromatic ring atom is carbon. In some embodiments, each ring atom is carbon. In some embodiments, the aryl group is biaryl. The term "aryl" may be used interchangeably with the term "aryl ring." In some embodiments, "aryl" refers to an aromatic ring system including phenyl, biphenyl, naphthyl, binatyl, anthracene, etc., which may have one or more substituents. As used herein, the term "aryl" also includes groups in which an aromatic ring is fused with one or more non-aromatic rings, such as indanyl, phthalimide, naphthylmethyl, phenanthridinyl, or tetrahydronaphthyl.

[0030] Halogen: The term "halogen" refers to F, Cl, Br or I.

[0031] Heteroatom: As used herein, the term "heteroatom" means an atom that is not carbon or hydrogen. In some embodiments, the heteroatom is boron, oxygen, sulfur, nitrogen, phosphorus, or silicon (including oxidized forms of nitrogen, sulfur, phosphorus, or silicon; charged forms of nitrogen, phosphorus, sulfur, or oxygen (e.g., quaternized forms, such as those in imine groups, etc.); etc.). In some embodiments, the heteroatom is silicon, phosphorus, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is silicon, oxygen, sulfur, or nitrogen. In some embodiments, the heteroatom is oxygen, sulfur, or nitrogen.

[0032] Partially unsaturated: As used herein, the term “partially unsaturated” means a ring moiety comprising at least one double or triple bond. The term “partially unsaturated” as used herein is intended to cover rings having multiple unsaturated sites, but is not intended to include aryl or heteroaryl moiety as defined herein.

[0033] Unsaturated: As used in this article, the term “unsaturated” means that a portion of a part has one or more unsaturated units.

[0034] Heteroaryl: As used herein, the terms “heteroaryl” and “heteroaryl-” used alone or as a larger part such as “heteroarylalkyl” or “heteroarylalkoxy” refer to, unless otherwise specified, a monocyclic, bicyclic, or polycyclic system having a total of 5 to 30 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 5-25, 5-20, 5-15, 5-14, 5-10, 5-9, etc.) ring members, wherein at least one ring in the system is aromatic and at least one aromatic ring atom is a heteroatom. In some embodiments, the number of cyclic heteroatoms in the heteroaryl group is about 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.). In some embodiments, the heteroaryl group is a group having 5 to 10 ring atoms, and in some embodiments 5, 6, 9, or 10 ring atoms. In some embodiments, each monocyclic unit is aromatic. In some embodiments, each aromatic monocyclic unit is independently 5- or 6-membered. In some embodiments, the heteroaryl ring is monocyclic and 5-membered. In some embodiments, the heteroaryl ring is monocyclic and 6-membered. In some embodiments, the heteroaryl ring is bicyclic and 9-membered. In some embodiments, the heteroaryl ring is bicyclic and 10-membered. In some embodiments, the heteroaryl group has 6, 10, or 14 π electrons shared in the cyclic array; and has one to five heteroatoms in addition to carbon atoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. Heteroaryl groups include, but are not limited to, thiophene, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, indoleazinyl, purine, naphridinyl, and pteridinyl. In some embodiments, the heteroaryl group is a heterobiaryl group, such as bipyridinyl. The terms "heteroaryl" and "heteroaryl-" as used herein also include groups in which a heteroaryl ring is fused with one or more aryl, alicyclic, or heterocyclic groups, wherein the linking group or linking point is on the heteroaryl ring. Non-limiting examples include indolyl, isoindolyl, benzothiophenyl, benzofuranyl, dibenzofuranyl, indazoleyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, cyclolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinazinyl, carbazoleyl, acridineyl, phenazinyl, phenothiazinyl, phenotoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups can be monocyclic, bicyclic, or polycyclic. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl," or "heteroarylene," any of which includes optionally substituted rings.The term "heteroaryl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are optionally substituted independently.

[0035] Furthermore, it should be understood that when two groups cyclize to form an optionally substituted heteroaryl ring having at least one nitrogen atom, the nitrogen atom in the ring can be (provided the valence state allows) N or NR. † As defined in this article.

[0036] Heterocycle: As used herein, the terms “heterocycle,” “heterocyclic group,” “heterocyclic ring,” and “heterocyclic ring” are used interchangeably as used herein and refer to a monocyclic, bicyclic, or polycyclic portion (unless otherwise specified, 3-30 members (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 3-25, 3-20, 3-15, 3-14, 3-10, 5-10, 3-7, 3-6, etc.)) that is saturated or partially unsaturated and has one or more (e.g., 1-10, 1-5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) heterocyclic ring atoms. In some embodiments, the number of cyclic heteroatoms in the heterocyclic group is about 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.). In some embodiments, the heterocyclic group is a stable 3-7 membered monocyclic or 7-10 membered bicyclic heterocyclic portion that is saturated or partially unsaturated and has one or more, preferably one to four, heteroatoms as defined above, in addition to a carbon atom. In some embodiments, each monocyclic group is independently 3-10 membered (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 3-8, 3-7, 3-6, 4-6, 5-6, 5-8, etc.). In some embodiments, each monocyclic ring independently has 0-5 heteroatoms (e.g., 0, 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.), wherein at least one monocyclic ring independently has 1-5 heteroatoms (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.). In some embodiments, each monocyclic ring is independently 3-7-membered. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. When referring to the ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, and nitrogen, nitrogen can be N (e.g., in 3,4-dihydro-2H-pyrroleyl), NH (e.g., in pyrroleyl), or... +NR (as in N-substituted pyrrolidinyl groups). Heterocycles may be attached to their side groups at any heteroatom or carbon atom that produces a stable structure, and any ring atom may optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolyl, piperazine, dioxalyl, dioxopentyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms “heterocyclic,” “heterocyclyl,” “heterocyclic ring,” “heterocyclic group,” “heterocyclic moiety,” and “heterocyclic group” are used interchangeably herein and also include groups in which the heterocyclic ring is fused with one or more aryl, heteroaryl, or alicyclic rings, such as dihydroindolyl, 3H-indolyl, chromanyl, phenanthridineyl, or tetrahydroquinolinyl. Heterocyclic groups can be monocyclic, bicyclic, or polycyclic. The term “heterocyclic alkyl” refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl and heterocyclic moiety are optionally substituted independently.

[0037] Furthermore, it should be understood that when two groups cyclize to form an optionally substituted heterocyclic ring having at least one nitrogen atom, the nitrogen atom in the ring can be (provided the valence state allows) N or NR. † As defined below.

[0038] Optionally substituted: As described herein, compounds of this disclosure may contain optionally substituted, substituted, and / or unsubstituted moieties. Generally, the term "substituted" means that one or more hydrogens of the specified moiety are independently substituted by a substituent. Unless otherwise stated, an "optionally substituted" group may have a substituent independently at each substituted position of the group, and when more than one position in any given structure can be substituted by two or more substituents, the substituents at each position may be the same or different. In some embodiments, the optionally substituted group is unsubstituted. In some embodiments, the optionally substituted group is substituted. Various substituents are described below.

[0039] The monovalent substituent is independently a halogen; -(CH2) 0-4 R°;-(CH2) 0-4 OR°;-O(CH2) 0-4 R°、-O-(CH2) 0-4 C(O)OR;-(CH2) 0-4 CH(OR°)2;-(CH2) 0-4Ph, which can be replaced by R; -(CH2) 0-4 O(CH2) 0-1 Ph, which can be substituted by R; -CH=CHPh, which can be substituted by R; -(CH2) 0-4 O(CH2) 0-1 -pyridyl group, which can be substituted with R; -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R°)2;-(CH2) 0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0-4 N(R°)C(O)N(R°)2;-N(R°)C(S)N(R°)2;-(CH2) 0-4 N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R°)N(R°)C(O)N(R°)2; -N(R°)N(R°)C(O)OR°; -(CH2) 0-4 C(O)R°;-C(S)R°;-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 C(O)SR°;-(CH2) 0-4 C(O)OSi(R°)3;-(CH2) 0-4 OC(O)R°;-OC(O)(CH2) 0-4 SR, -SC(S)SR; -(CH2) 0-4 SC(O)R°;-(CH2) 0-4 C(O)N(R°)2; -C(S)N(R°)2; -C(S)SR; -SC(S)SR, -(CH2) 0-4 OC(O)N(R°)2;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH2C(O)R°;-C(NOR°)R°;-(CH2) 0-4 SSR°;-(CH2) 0-4 S(O)2R°;-(CH2) 0-4 S(O)₂OR°;-(CH₂) 0-4 OS(O)2R°;-S(O)2N(R°)2;-(CH2) 0-4S(O)R°; -N(R°)S(O)2N(R°)2; -N(R°)S(O)2R°; -N(OR°)R°; -C(NH)N(R°)2; -Si(R°)3; -OSi(R°)3; -P(R°)2; -P(OR °)2; -OP(R°)2; -OP(OR°)2; -N(R°)P(R°)2; -B(R°)2; -OB(R°)2; -P(O)(R°)2; -OP(O)(R°)2; -N(R°)P(O)(R°)2; -(C 1-4 (linear or branched alkylene)ON(R°)2; or -(C 1-4 (straight-chain or branched alkylene)C(O)ON(R°)2; wherein each R° can be independently substituted as defined below, and is independently hydrogen, C 1-10 (For example, C) 1-6 C 1-5 C 1-4 Aliphatic carbon atoms having 1-5 heteroatoms (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus. 1-10 (For example, C) 1-6 C 1-5 C 1-4 (etc.) mixed lipids, C 6-10 (For example, C6, C) 10 aryl groups, 5-10 heteroaryl groups having 1-4 heteroatoms (e.g., 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) independently selected from nitrogen, oxygen, and sulfur, -CH2-(C 6-10 (For example, C6, C) 10 Aryl group, -O(CH2) 0-1 (C 6-10 (For example, C6, C) 10 -(aryl), -CH2- (5-10 membered (e.g., 5-9, 5-6, 5, 6, 9, 10 membered, etc.) heteroaryl groups having 1-4 (e.g., 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen and sulfur), -O(CH2). 0-1(Having 1-4 heteroaryl groups (e.g., 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) independently selected from nitrogen, oxygen, and sulfur, consisting of 5-10 membered heteroaryl groups (e.g., 5-9, 5-6, 5, 6, 9, 10, etc.), having 0-5 heteroaryl groups (e.g., 0, 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, consisting of 3-10 membered heteroaryl groups (e.g., 3-6, 5-6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) monocyclic, bicyclic, or polycyclic, saturated or partially unsaturated rings, or, despite the above. By definition, two independent occurrences of R° together with their intermediate atoms form 3-10 cyclic (e.g., 3-6, 5-6, 3, 4, 5, 6, 7, 8, 9, 10, etc.) monocyclic, bicyclic, or polycyclic, saturated, partially unsaturated, or aromatic rings (for aromatic rings, 5-10 cyclic (e.g., 5-9, 5-6, 5, 6, 9, 10, etc.)). In addition to the intermediate atoms, the ring also has 0-5 heteroatoms (e.g., 0, 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) independently selected from nitrogen, oxygen, sulfur, silicon, and phosphorus, which can be substituted as defined below.

[0040] The monovalent substituents on R° (or the ring formed by the two independent occurrences of R° along with the added atoms) are independently halogens, -(CH2). 0-2 R ● -(halogenated R) ● -(CH2) 0-2 OH, -(CH2) 0-2 OR ● -(CH2) 0-2 CH(OR ● )2;-O(halogenated R ● -CN, -N3, -(CH2) 0-2 C(O)R ● -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● -(CH2) 0-2 SR ● -(CH2) 0- 2SH、-(CH2) 0-2 NH2、-(CH2) 0-2 NHR ● -(CH2) 0-2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● -(C 1-4 (straight-chain or branched alkylene)C(O)OR● or -SSR ● , where each R ● It is unsubstituted or, if preceded by a "halogen group," substituted with only one or more halogens, and independently selected from C. 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or has 0-4 (e.g., 0, 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, and 3-6 membered (e.g., 3, 4, 5, 6, 3-5, 5-6 membered, etc.) saturated, partially unsaturated, or aromatic rings (for aromatic rings, 5 or 6 membered). The divalent substituents on the saturated carbon atom of R° are independently =O or =S.

[0041] The divalent substituents are independently the following: =O, =S, =NNR * 2、=NNHC(O)R * =NNHC(O)OR * =NNHS(O)2R * =NR * =NOR * -O(C(R) * 2)) 2-3 O- or -S(C(R) * 2)) 2-3 S-, where R * Each independent occurrence of is selected from hydrogen, and can be substituted by C as defined below. 1-6 Aliphatic or unsubstituted 3-6 membered (e.g., 3, 4, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, of a saturated, partially unsaturated, or aromatic ring (for aromatic rings, 5 or 6 membered). The divalent substituent bonded to the ortho-substituted carbon of the "optionally substituted" group is independently -O(CR). * 2) 2-3 O-, where R * Each independent occurrence of is selected from hydrogen, and can be substituted by C as defined below. 1-6 Aliphatic or having 0-4 (e.g., 0, 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) unsubstituted 3-6 membered (e.g., 3, 4, 5, 6, 3-5, 5-6 membered, etc.) heteroatoms independently selected from nitrogen, oxygen and sulfur, saturated, partially unsaturated or aromatic rings (for aromatic rings, 5 or 6 membered).

[0042] R * The substituents on the aliphatic group are independently halogens, -R ● -(halogenated R) ● -OH, -OR● -O(halogenated R) ● -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● -NR ● 2 or -NO2, where each R ● It is either unsubstituted or, when preceded by a "halogen group," substituted with only one or more halogens, and is independently C. 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 3-6 member (e.g., 3, 4, 5, 6, 3-5, 5-6, etc.) saturated, partially unsaturated or aromatic ring (for aromatic rings, 5 or 6 members) having 0-4 (e.g., 0, 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0043] The substituents on the substituted nitrogen are independently -R † -NR † 2. -C(O)R † -C(O)OR † -C(O)C(O)R † -C(O)CH2C(O)R † -S(O)2R † -S(O)2NR † 2. -C(S)NR † 2. -C(NH)NR † 2 or -N(R) † )S(O)2R † ; where each R † Independently hydrogen, C can be substituted as defined below. 1-6 Aliphatic, unsubstituted -OPh or unsubstituted 3-6 membered (e.g., 3, 4, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, saturated, partially unsaturated, or aromatic rings (5 or 6 members for aromatic rings), or, despite the above definition, R † The two independent occurrences of these atoms together with their intermediate atoms form unsubstituted 3-12-membered (e.g., 3-10, 3-6, 5-10, 5-6, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen and sulfur, saturated, partially unsaturated or aryl monocyclic or bicyclic rings.

[0044] R † The substituents on the aliphatic group are independently halogens, -R● -(halogenated R) ● -OH, -OR ● -O(halogenated R) ● -CN, -C(O)OH, -C(O)OR ● -NH2, -NHR ● -NR ● 2 or -NO2, where each R ● It is either unsubstituted or, when preceded by a "halogen group," substituted with only one or more halogens, and is independently C. 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph or a 3-6 member (e.g., 3, 4, 5, 6, 3-5, 5-6, etc.) saturated, partially unsaturated or aromatic ring (for aromatic rings, 5 or 6 members) having 0-4 (e.g., 0, 1, 2, 3, 4, 1-4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0045] Pharmaceutically acceptable salts: As used herein, the term "pharmaceutically acceptable salt" means those salts which, to a reasonable extent of medical judgment, are suitable for contact with tissues of humans and lower animals without excessive toxicity, irritation, anaphylactic response, etc., and which are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19 (incorporated by reference). Pharmaceutically acceptable salts include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by amino groups with inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or salts formed by other methods used in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, disglucuronate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptahydrate, glyceryl phosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc.

[0046] Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and [N(C)] salts. 1-4 [alkyl)4] + Salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts include, where appropriate, non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates.

[0047] Unless otherwise stated, the structures described herein are intended to include all isomers (e.g., enantiomers, diastereomers, and geometric (or conformations)) of the structures; for example, R and S configurations for each asymmetric center, Z and E double bond isomers, etc. Therefore, unless otherwise stated, single stereochemical isomers of the compounds of the present invention, as well as mixtures of enantiomers, diastereomers, and geometric (or conformations), are within the scope of this disclosure. Unless otherwise stated, all tautomers are within the scope of this disclosure. As will be understood by those skilled in the art, compounds and elements can exist in various isotopic forms. Unless otherwise stated, this disclosure includes compounds that differ only in the presence of one or more isotopes at one or more sites, and compositions of compounds with isotopic enrichment at one or more sites. For example, unless otherwise stated, in a compound, H can be... 1 H, 2 H or 3 H, and C can be 12 C 13 C or 14 C. In some embodiments, compounds having certain isotopes or compositions enriched with certain isotopes may be used, for example, as analytical tools, as probes in bioassays, or as therapeutic agents according to this disclosure. In some embodiments, the compounds of this disclosure contain one or more deuterium atoms. In some embodiments, the provided agents, compounds, conjugates, etc., are in salt form, such as pharmaceutically acceptable salt forms.

[0048] In some embodiments, the combinations of substituents and variables contemplated in this disclosure are only those that result in the formation of stable compounds, where “stable” means that the compound has sufficient stability to allow manufacture and maintains the integrity of the compound for a sufficiently long time to be used for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0049] Other definitions Approximately: As used herein, the term “approximately” applied to one or more values ​​of interest means a value similar to the stated reference value. In some embodiments, unless otherwise stated or clearly apparent from the context (except where such a number would exceed 100% of the possible value), the term “approximately” means a range of values ​​falling within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of the stated reference value.

[0050] Affinity maturation: As used herein, the term "affinity maturation" refers to an antibody having one or more alterations in one or more of its CDRs, which result in improved antibody affinity for the antigen compared to a parent antibody without the alterations. In some embodiments, the affinity-matured antibody has nanomolar or even picomolar affinity for the target antigen. Affinity-matured antibodies can be produced by any of a variety of procedures known in the art. Affinity maturation achieved through VH and VL domain shuffling is described in Marks et al., BioTechnology 10:779-783 (1992). Random mutagenesis of CDRs and / or framework residues is described in the following literature: Barbas et al., Proc. Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al., Gene 169: 147-155 (1995); Yelton et al., J. Immunol. 155: 1994-2004 (1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins et al., J. Mol. Biol. 226:889-896 (1992).

[0051] Agent: As used herein, the term "agent" refers to a biological entity and / or compound, including, for example, antibodies or antigen-binding fragments thereof, organic molecules (e.g., small molecules), peptides (e.g., fusion proteins), aptamers, nucleic acids, chimeric antigen receptors, glycoproteins, sugars, lipids, growth factors, enzymes, synthetic molecules, carbohydrates, lipids, hormones, polymers or derivatives thereof, variants, complexes, or any combination thereof. Where appropriate, as will be clear from the context to a person skilled in the art, the term may be used to refer to an entity that is or includes a cell or organism or a fraction, extract, or component thereof. Alternatively or additionally, as will be clear from the context, the term may be used to refer to a natural product. In some cases, also as will be clear from the context, the term may be used to refer to one or more artificial entities because it is designed, engineered, and / or produced by human action and / or is not found in nature. In some embodiments, the agent may be used in isolated or pure form. In some embodiments, the agent may be used in crude form. In some embodiments, the agent is provided as a collection or library that may be screened to identify or characterize the active agents therein. The agent can bind to any cellular part, such as receptors, antigenic determinants, or other binding sites present on the target or target cell. Various agents can be used in the compositions and methods described herein.

[0052] Antibody: As used herein, the term "antibody" refers to a polypeptide comprising typical immunoglobulin sequence elements sufficient to confer specific binding to a particular target antigen. As is known in the art, a complete antibody, such as those produced in nature, is a tetramer of approximately 150 kD, comprising two identical heavy-chain polypeptides (each approximately 50 kD) and two identical light-chain polypeptides (each approximately 25 kD), which associate with each other to form a structure commonly referred to as a "Y-shape." Each heavy chain contains at least four domains (each approximately 110 amino acids in length)—an amino-terminal variable (VH) domain (located at the apex of the Y structure), followed by three constant domains: CH1, CH2, and a carboxyl-terminal CH3 (located at the base of the stem of the Y). A short region called a "switch" connects the heavy-chain variable and constant regions. A "hinge" connects the CH2 and CH3 domains to the rest of the antibody. Two disulfide bonds in this hinge region link the two heavy-chain polypeptides in the complete antibody to each other. Each light chain contains two domains—an amino-terminal variable (VL) domain and a carboxyl-terminal constant (CL) domain, separated from each other by another “switch.” A complete antibody tetramer consists of two heavy-chain-light-chain dimers, where the heavy and light chains are linked together by a single disulfide bond; two additional disulfide bonds link the heavy-chain hinge regions together, causing the dimers to link together and form a tetramer. Naturally occurring antibodies are typically also glycosylated at the CH2 domain. Each domain in a natural antibody has a structure characterized by an “immunoglobulin fold,” formed by two β-sheets (e.g., 3-, 4-, or 5-folds) stacked on top of each other in compressed antiparallel β-barrels. Each variable domain contains three hypervariable loops called “complementarity-determining regions” (CDR1, CDR2, and CDR3) and four slightly invariant “framework” regions (FR1, FR2, FR3, and FR4). When a natural antibody folds, the FR region forms a β-sheet that provides a structural framework for the domain, and the CDR loop regions from both the heavy and light chains aggregate in three-dimensional space, creating a single hypervariable antigen-binding site at the apex of the Y-structure. The Fc region of a naturally occurring antibody binds to elements of the complement system and also to receptors on effector cells, including, for example, effector cells mediating cytotoxicity. The affinity and / or other binding properties of the Fc region to Fc receptors can be modulated by glycosylation or other modifications. In some embodiments, antibodies generated and / or utilized according to this disclosure include glycosylated Fc domains, such as those having modified or engineered glycosylated Fc domains. In some embodiments, any polypeptide or polypeptide complex containing sufficient immunoglobulin domain sequences as found in natural antibodies can be referred to as and / or used as an "antibody," regardless of whether such polypeptide is naturally generated (e.g., produced by an organism's response to an antigen) or generated by recombinant engineering, chemical synthesis, or other artificial systems or methods.In some embodiments, the antibody is polyclonal. In some embodiments, the antibody is monoclonal. In some embodiments, the antibody has a constant region sequence specific to mouse, rabbit, primate, or human antibodies. In some embodiments, the antibody sequence element is humanized, primate-derived, or chimeric, as is known in the art. Furthermore, the term "antibody" as used herein may, in appropriate embodiments (unless otherwise stated or clearly apparent from the context), refer to any construct or form known or developed in the art that utilizes the structural and functional characteristics of an antibody in an alternative presentation. For example, in some embodiments, the antibody utilized according to the provided technology is selected from, but not limited to, intact IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., Zybodies®, etc.); and / or antibody fragments (preferably antibody fragments exhibiting the desired antigen-binding activity). The antibodies described herein can be immunoglobulins, heavy chain antibodies, light chain antibodies, LRR-based antibodies, or other protein scaffolds with antibody-like properties, as well as any other immune-binding motif known in the art, such as Fab, Fab', Fab'2, Fab2, Fab3, F(ab')2, Fd, Fv, Feb, scFv, SMIP, antibodies, biantibodies, triantibodies, tetraantibodies, microantibodies, large antibodies, tandab, DVD, BiTe, TandAb, or any combination thereof. The subunit structures and three-dimensional configurations of different types of antibodies are known in the art. In some embodiments, the antibody may lack covalent modifications (e.g., glycan linkages) that it would have naturally. In some embodiments, the antibody may contain covalent modifications (e.g., glycan linkages), payloads (e.g., detectable portions, therapeutic portions, catalytic portions, etc.) or other side groups (e.g., polyethylene glycol, etc.).

[0053] Antibody Agent: As used herein, the term "antibody agent" refers to an agent that specifically binds to a particular antigen. In some embodiments, the term covers any polypeptide or polypeptide complex that includes sufficient immunoglobulin structural elements to confer specific binding. Exemplary antibody agents include, but are not limited to, monoclonal or polyclonal antibodies. In some embodiments, an antibody agent may include one or more constant region sequences having characteristics of mouse, rabbit, primate, or human antibodies. In some embodiments, an antibody agent may include one or more humanized, primate-based, or chimeric sequence elements, as known in the art. In many embodiments, the term "antibody agent" is used to refer to one or more constructs or forms known or developed in the art for utilizing the structural and functional characteristics of an antibody in an alternative presentation. For example, the antibody agents utilized according to the provided technology are, but are not limited to, those resembling: intact IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., Zybodies®); antibody fragments, such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated CDRs or collections thereof; single-chain Fv; peptide-Fc fusions; single-domain antibodies (e.g., shark single-domain antibodies, such as IgNAR or fragments thereof); camelid antibodies; masking antibodies (e.g., Probodies®); Small Modular ImmunoPharmaceuticals (“SMIP™”). ”Single-chain or tandem biantibodies (TandAb®); VHH; Anticalins®; Nanobodies® microantibodies; BiTE®; Ankylosing spicules or DARPINs®; Avimers®; DART; TCR-like antibodies; Adnectins®; Affilins®; Trans-bodies®; Affibodies®; TrimerX®; Microbial proteins; Fynomers®; Centyrins®; or KALBITOR®. In some embodiments, the antibody lacks the covalent modifications (e.g., glycan linkages) that it would have naturally. In some embodiments, the antibody contains covalent modifications (e.g., glycan linkages, payloads such as detectable, therapeutic, or catalytic moieties) or other side groups (e.g., polyethylene glycol). In many embodiments, the antibody agent is or includes a polypeptide whose amino acid sequence includes one or more structural elements recognized by those skilled in the art as complementarity-determining regions (CDRs); in some embodiments, the antibody agent is or includes a polypeptide whose amino acid sequence includes at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) substantially identical to the CDR found in a reference antibody. In some embodiments, the included CDR is substantially identical to the reference CDR because it is sequence-identical or contains 1-5 amino acid substitutions compared to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR, having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR, having at least 96%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because at least one amino acid in the included CDR is deleted, added, or substituted compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because 1-5 amino acids in the included CDR are deleted, added, or substituted compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because at least one amino acid in the included CDR is substituted compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the reference CDR.In some embodiments, the included CDR is substantially identical to the reference CDR because 1-5 amino acids are deleted, added, or substituted compared to the reference CDR, but the included CDR has the same amino acid sequence as the reference CDR in all other respects. In some embodiments, the antibody agent is or includes a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as immunoglobulin variable domains. In some embodiments, the antibody agent is a polypeptide protein having a binding domain homologous or substantially homologous to an immunoglobulin binding domain.

[0054] Antibody heavy chain: As used herein, the term “antibody heavy chain” refers to the larger of the two types of polypeptide chains that are present in all antibodies in their naturally occurring conformation.

[0055] Antibody light chain: As used herein, the term “antibody light chain” refers to the smaller of the two types of polypeptide chains that exist in all antibodies in their naturally occurring conformation.

[0056] Antigen: As used herein, the term "antigen" or "Ag" refers to a molecule capable of eliciting an immune response. This immune response may involve antibody production, activation of specific immune cells, or both. Those skilled in the art will understand that any macromolecule, including virtually all proteins or peptides, can be used as an antigen. Furthermore, antigens can be derived from recombinant or genomic DNA. Those skilled in the art will understand that any DNA containing a nucleotide sequence or a portion of a protein encoding an immune response encodes the term "antigen" as used herein. Furthermore, those skilled in the art will understand that an antigen need not be encoded solely by the full-length nucleotide sequence of a gene. It is apparent that the techniques provided include, but are not limited to, the use of partial nucleotide sequences of more than one gene, and that these nucleotide sequences are arranged in various combinations to elicit the desired immune response. Furthermore, those skilled in the art will understand that an antigen need not be encoded by a "gene" at all. It is apparent that antigens can be synthesized or can be derived from biological samples. Such biological samples may include, but are not limited to, tissue samples, tumor samples, cells, or biological fluids.

[0057] Antigen-binding fragment: As used herein, the term "antigen-binding fragment" refers to a portion of a complete antibody that binds to the antigen bound by the complete antibody. Antigen-binding fragments of antibodies include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Exemplary antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; biantibodies; linear antibodies; single-chain antibody molecules (e.g., scFv, VHH, camel antibody, or VH or VL domain only); or multispecific antibodies formed from antibody fragments. In some embodiments, the antigen-binding fragment of the antibody described herein is scFv. In some embodiments, the antigen-binding fragment of the antibody described herein is only the VHH domain. Like the whole antibody molecule, the antigen-binding fragment can be monospecific or multispecific (e.g., bispecific). A multispecific antigen-binding fragment of an antibody may contain at least two distinct variable domains, each of which is capable of specifically binding to a single antigen or different epitopes of the same antigen. Antigen-binding fragments can be generated in any manner. For example, in some embodiments, the antigen-binding fragment is produced enzymatically or chemically by fragmentation of a complete antibody or antibody agent. Alternatively, in some embodiments, the antigen-binding fragment is recombinantly generated. In some embodiments, the antigen-binding fragment is synthesized wholly or partially. In some embodiments, the antigen-binding fragment has a length of at least about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 amino acids or greater.

[0058] Antibody-dependent cytotoxicity: As used herein, the term “antibody-dependent cytotoxicity” or “ADCC” refers to the phenomenon in which target cells bound to antibodies are killed by immune effector cells. Without being bound by theory, ADCC is generally understood to involve effector cells carrying Fc receptors (FcRs) recognizing and subsequently killing antibody-coated target cells (e.g., cells expressing specific antigens on their surface that bind to antibodies). Effector cells mediating ADCC include immune cells, including but not limited to natural killer (NK) cells, macrophages, neutrophils, and eosinophils.

[0059] Aptamers: As used herein, the term "aptamer" refers to a macromolecule composed of nucleic acids (e.g., RNA, DNA) that binds tightly to a specific molecular target (e.g., umbrella-shaped topoglycans). Specific aptamers can be described by linear nucleotide sequences and are typically about 15–60 nucleotides in length. Without wishing to be bound by any theory, it is envisioned that the nucleotide chains in the aptamer form intramolecular interactions that fold the molecule into a complex three-dimensional shape, and that this three-dimensional shape allows the aptamer to bind tightly to the surface of its target molecule. Given the extraordinary diversity of molecular shapes present across all possible nucleotide sequence categories, aptamers are available for a wide array of molecular targets, including proteins and small molecules. In addition to high specificity, aptamers typically exhibit very high affinity for their targets (e.g., affinity for proteins in the picomolar to low nanomolar range). In many embodiments, aptamers are chemically stable and can be boiled or frozen without losing their activity. As they are synthetic molecules, aptamers can be modified in various ways, which can optimize their function for a specific application. For example, aptamers can be modified to significantly reduce their sensitivity to enzymatic degradation in blood used in in vivo applications. Furthermore, aptamers can be modified to alter their biodistribution or plasma residence time.

[0060] Related: Two events or entities are "related" to each other if the presence, level, and / or form of one is associated with the presence, level, extent, type, and / or form of the other. For example, if the presence, level, and / or form of a particular entity (e.g., a polypeptide, genetic marker, metabolite, or microorganism) is associated with the incidence and / or susceptibility to a disease, symptom, or condition (e.g., in a relevant population), that particular entity is considered related to that particular disease, symptom, or condition. In some embodiments, if two or more entities interact directly or indirectly, they are physically "related" to each other such that they are physically close to each other and / or remain physically close to each other. In some embodiments, two or more physically related entities are covalently connected to each other. In some embodiments, two or more physically related entities are not covalently connected to each other but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0061] Binding: As used herein, the term “binding” refers to a non-covalent association between or among two or more entities. “Direct” binding involves physical contact between entities or parts. Indirect binding involves physical interaction achieved through physical contact with one or more intermediate entities. Binding between two or more entities can generally be evaluated in any of a variety of cases, including studying the interacting entities or parts individually or in the case of more complex systems (e.g., when covalently associated with a carrier entity or otherwise associated and / or in a biological system or cell).

[0062] Cancer: As used herein, the terms “cancer,” “malignant tumor,” “vesicle,” “tumor,” and “carcinoma” refer to cells that exhibit relatively abnormal, uncontrolled, and / or autonomous growth, thus displaying an abnormal growth phenotype characterized by a significant loss of control over cell proliferation. Cancer cells can spread locally or to other parts of the body via the bloodstream and lymphatic system. In some embodiments, a tumor is or includes precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and / or non-metastatic cells. In some embodiments, cancer is or includes a solid tumor. In some embodiments, cancer is or includes a hematologic malignancy. Examples of various cancers described herein, including but not limited to, cancers of the hematopoietic system, including leukemia, lymphoma (Hodgkin's lymphoma and non-Hodgkin's lymphoma), myeloma, and myeloproliferative disorders; sarcoma, melanoma, adenoma, solid tissue cancer, squamous cell carcinoma of the oral cavity, pharynx, larynx, and lungs, liver cancer, cancers of the genitourinary system (such as prostate cancer, cervical cancer, bladder cancer, uterine cancer, endometrial cancer, and renal cell carcinoma), bone cancer, pancreatic cancer, skin cancer, melanoma of the skin or eye, cancers of the endocrine system, thyroid cancer, parathyroid cancer, head and neck cancer, breast cancer, gastrointestinal cancer, cancers or benign lesions of the nervous system such as papilloma, and several other types, including those described elsewhere herein.

[0063] Carrier: As used herein, "carrier" refers to a diluent, adjuvant, excipient, and / or medium applied together with the composition. In some exemplary embodiments, the carrier includes sterile liquids, such as water and oils, including petroleum, animal, plant, or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. In some embodiments, the carrier is or comprises one or more solid components.

[0064] CDR: As used herein, “CDR” refers to the complementarity-determining region within the antibody variable region. Three CDRs exist in each variable region of the heavy and light chains, designated as CDR1, CDR2, and CDR3 for each variable region. A “group of CDRs” or “CDR set” refers to a group of three or six CDRs appearing in a single variable region capable of binding the antigen, or in CDRs of homologous heavy and light chain variable regions capable of binding the antigen. The precisely defined boundaries and lengths of CDRs depend on different classification and numbering systems. Certain systems have been established in the art for defining CDR boundaries (e.g., Kabat, IMGT, Chothia, or combinations thereof). Therefore, a CDR can be referred to by Kabat, Chothia, IMGT, or any other boundary definition known in the art. Despite the different boundaries, each of these systems has a degree of overlap in terms of the “hypervariate regions” that constitute the variable sequence. Therefore, the CDR definitions of these systems may differ in length and boundary regions relative to adjacent frame regions (see, for example, Kabat et al., “Sequences of Proteins of Immunological Interest,” 5th ed., US Department of Health and Human Services, 1992; Chothia et al. (1987) J. Mol. Biol. 196, 901; and MacCallum et al., J. Mol. Biol. (1996) 262, 732, each of which is incorporated herein by reference in its entirety). Those skilled in the art will understand the differences between these systems and will be able to grasp the CDR boundaries to the extent necessary to understand and practice the claims and disclosure herein.

[0065] Chemotherapy agents: As used herein, the term "chemotherapy agent" has its meaning as understood in the art, referring to one or more pro-apoptotic agents, cell inhibitors, and / or cytotoxic agents, specifically including agents used and / or recommended for the treatment of one or more diseases, conditions, or illnesses associated with unwanted cell proliferation. In many embodiments, chemotherapy agents can be used to treat cancer. In some embodiments, chemotherapy agents may be or comprise one or more alkylating agents, one or more anthracycline drugs, one or more cytoskeleton disruptors (e.g., microtubule-targeting agents such as taxanes, maytansine, and analogues thereof), one or more epothilosomatic agents, one or more histone deacetylase inhibitors (HDAC), one or more topoisomerase inhibitors (e.g., inhibitors of topoisomerase I and / or topoisomerase II), one or more kinase inhibitors, one or more nucleotide analogs or nucleotide precursor analogs, one or more peptide antibiotics, one or more platinum-based agents, one or more vitamin A-like agents, one or more vinca alkaloids, and / or one or more analogues of one or more of the following (i.e., sharing associated antiproliferative activity).In some implementations, the chemotherapeutic agent may be one or more of the following: actinomycin, all-trans retinoic acid, auristatin, azacitidine, azathioprine, bleomycin, bortezomib, carboplatin, capecitabine, cisplatin, chlorambucil. Cyclophosphamide, curcumin, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, etoposide, fluorouracil, gemcitabine Gemcitabine, Hydroxyurea, Idarubicin, Imatinib, Irinotecan, Maytansine, and / or their analogues (e.g., DM1), Mechlorethamine, Mercaptopurine, Methotrexate, Mitoxantrone, Maytansinoid, Oxaliplatin, Paclitaxel, Pemetrexed, Teniposide, Tioguanine, Topotecan, Valrubicin, Vinblastine, Vincristine, Vindesine, Vinorelbine, or combinations thereof. In some implementations, chemotherapeutic agents can be used in the context of antibody-drug conjugates.In some implementations, the chemotherapeutic agent is an antibody-drug conjugate comprising: hLL1-doxorubicin, hRS7-SN-38, hMN-14-SN-38, hLL2-SN-38, hA20-SN-38, hPAM4-SN-38, hLL1-SN-38, hRS7-Pro-2-P-Dox, hMN-14-Pro-2-P-Dox, hLL2-Pro-2-P-Dox, hA20-Pro-2-P-Dox, hPAM4-Pro-2-P-Dox, hLL1-Pro-2-P-Dox, P4 / D10-doxorubicin, gemtuzumab ozogamicin, brentuximab vedotin, and trastuzumab vedotin. emtansine), intuzumab octozomicin, glembatumomab vedotin, SAR3419, SAR566658, BIIB015, BT062, SGN-75, SGN-CD19A, AMG-172, AMG-595, BAY-94-9343, ASG-5ME, ASG-22ME, ASG-16M8F, MDX-1203, MLN-0264, anti-PSMA ADC, RG-7450, RG-7458, RG-7593, RG-7596, RG-7598, RG-7599, RG-7600, RG-7636, ABT-414, IMGN-853, IMGN-529, vorsetuzumab mafodotin, and lorvotuzumab mertansine.

[0066] Chimeric antibody: As used herein, it refers to an antibody whose amino acid sequence comprises the VH and VL regions found in a first species and the constant region sequence found in a second species different from the first species. In many embodiments, chimeric antibodies have mouse VH and VL regions linked to human constant regions. In some embodiments, antibodies having human VH and VL regions linked to non-human constant regions (e.g., mouse constant regions) are referred to as “reverse chimeric antibodies.”

[0067] Composition: Those skilled in the art will understand that the term "composition" can be used to refer to a discrete physical entity comprising one or more specified components. Generally, unless otherwise specified, a composition can be in any form—for example, a gas, gel, liquid, or solid.

[0068] **Inclusion:** The description herein of a composition or method “comprising” one or more specified elements or steps is open-ended, meaning that the specified elements or steps are essential, but other elements or steps may be added within the scope of the composition or method. To avoid redundancy, it should also be understood that any composition or method described as “comprising” (or “including”) one or more specified elements or steps also describes a corresponding, more limited composition or method that “consists substantially of the same specified elements or steps” (or “forms substantially the same specified elements or steps”), meaning that the composition or method includes the specified essential elements or steps and may also include additional elements or steps that do not materially affect the essential and novel characteristics of the composition or method. It should also be understood that any composition or method described herein as “comprising” one or more specified elements or steps or “consists substantially of one or more specified elements or steps” also describes a corresponding, more limited and closed composition or method that “consists of the specified elements or steps” (or “forms substantially the specified elements or steps”) to exclude any other unspecified elements or steps. In any composition or method disclosed herein, any known or disclosed equivalent of a specified essential element or step may replace that element or step.

[0069] Conserved sequence modification: As used herein, the term "conserved sequence modification" refers to an amino acid modification that does not significantly affect or alter the binding properties of an antibody or its antigen-binding fragment containing that amino acid sequence. Such conserved modifications include amino acid substitutions, additions, and deletions. Modifications can be introduced into antibodies compatible with various embodiments using standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. A conserved amino acid substitution is an amino acid substitution in which an amino acid residue is replaced by an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with the following characteristics: basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), β-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Therefore, one or more amino acid residues within the CDR region of an antibody can be replaced by other amino acid residues from the same side chain family, and the altered antibody's ability to bind antigens can be tested using the functional assays described herein.

[0070] Combination therapy: As used herein, the term "combination therapy" refers to those situations in which two or more different therapeutic agents are administered in an overlapping regimen, thereby exposing a subject to both agents simultaneously. When used in combination therapy, the two or more different therapeutic agents may be administered simultaneously or separately. Such combination administration may include simultaneous administration of two or more therapeutic agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, two or more therapeutic agents may be formulated together in the same dosage form and administered simultaneously. Alternatively, two or more therapeutic agents may be administered simultaneously, wherein these therapeutic agents are present in separate formulations. In another alternative, a first therapeutic agent may be administered, followed by one or more additional therapeutic agents. In separate administration regimens, the two or more therapeutic agents may be administered at intervals of minutes, hours, days, or weeks. In some embodiments, the two or more therapeutic agents may be administered at intervals of hours (e.g., less than about 1 hour, about 2 hours, about 3 hours, about 4 hours, or about 5 hours).

[0071] Effective Amount: As used herein, an "effective amount" means a dose sufficient to prevent or treat at least one sign and / or symptom of a disease, condition, or illness (e.g., cancer) in an individual. The effective amount for therapeutic or preventative use will depend, for example, on the stage and severity of the disease, condition, or illness being treated, the patient's age, weight, and general health condition, and the judgment of the prescribing physician. The dosage will also be determined by the selected active substance, the method of administration, the time and frequency of administration, the presence, nature, and extent of any adverse side effects that may accompany the administration of the specific active substance, and the desired physiological effect. Those skilled in the art will understand that various diseases or conditions may require long-term treatment involving multiple administrations. For the purposes of this disclosure, the amount or dose of the therapeutic agent (e.g., at least one HHLA2 binder described herein) administered should be sufficient to achieve a therapeutic or preventative response (e.g., a reduction or other amelioration in the severity or duration of at least one sign or symptom) in a subject within a reasonable timeframe. For example, the dose should be sufficient to detect, treat, or prevent cancer for a period of approximately 2 hours or more, such as approximately 12 hours, to approximately 24 hours or more from the time of administration. In some implementations, this time period may be even longer. The dosage will be determined by the efficacy of one or more specific therapeutic agents and the condition of the subject (e.g., a person) and the weight of the subject to be treated (e.g., a person).

[0072] Encoding: As used herein, the term "encoding" refers to the inherent property of a specific nucleotide sequence in a polynucleotide (such as a gene, cDNA, or mRNA) that serves as a template for the synthesis of other polymers and macromolecules in biological processes, having a defined nucleotide sequence (e.g., rRNA, tRNA, or mRNA) or a defined amino acid sequence, and the resulting biological properties. Thus, if the transcription and translation of mRNA corresponding to a gene produces a protein in a cell or other biological system, then the gene encodes that protein. Both the coding strand, whose nucleotide sequence is identical to the mRNA sequence and is typically provided in the sequence listing, and the non-coding strand, which serves as a transcription template for a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0073] Engineered: As used herein, the term "engineered" refers to an aspect that has been artificially manipulated. For example, a polynucleotide is considered "engineered" when two or more sequences not linked together in a naturally occurring order are artificially manipulated to be directly linked to each other. For example, in some embodiments, an engineered polynucleotide contains, or is found in nature, a regulatory sequence that is operatively associated with a first coding sequence but not with a second coding sequence, which is artificially linked to make it operatively associated with the second coding sequence. In contrast, a cell or organism is considered "engineered" if it has been manipulated to alter its genetic information (e.g., new genetic material not previously present has been introduced, for example by transformation, mating, somatic cell hybridization, transfection, transduction, or other mechanisms, or previously existing genetic material has been altered or removed, for example by substitution or deletion mutations, or by mating protocols). In some embodiments, the engineered antibody or its antigen-binding fragment (e.g., the engineered monoclonal antibody or its antigen-binding fragment) includes VH and / or VL region sequences from a reference antibody produced in a non-human species (e.g., mouse) and modifications in these sequences relative to the reference antibody, which are intended to make them more “human-like” or more similar to human phylogenetic variable region sequences.

[0074] As is customary and understood by those skilled in the art, the offspring of engineered polynucleotides or cells are typically still referred to as “engineered,” even though the actual manipulation was performed on the prior entity.

[0075] Epitope: As used herein, the term "epitaph" refers to any portion specifically recognized by an immunoglobulin (e.g., antibody or receptor) binding component. In some embodiments, an epitope consists of multiple chemical atoms or groups on an antigen. In some embodiments, such chemical atoms or groups are surface-exposed when the antigen adopts an associated three-dimensional conformation. In some embodiments, such chemical atoms or groups are physically close to each other in space when the antigen adopts such a conformation. In some embodiments, at least some of such chemical atoms or groups are physically separated from each other when the antigen adopts an alternative conformation (e.g., linearization).

[0076] Expression: As used herein, the term “expression” of a nucleic acid sequence means the generation of any gene product from a nucleic acid sequence (e.g., a nucleic acid sequence encoding an anti-HHLA2 antibody or an antigen-binding fragment thereof as described herein). In some embodiments, the gene product may be a transcript. In some embodiments, the gene product may be a polypeptide. In some embodiments, the expression of a nucleic acid sequence involves one or more of the following: (1) generating an RNA template from a DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, 5' cap formation, and / or 3' end formation); (3) translating RNA into a polypeptide or protein; and / or (4) post-translational modification of the polypeptide or protein.

[0077] Segment: As used herein, the term "segment" refers to a structure that comprises a discrete part of the whole but lacks one or more parts found in the whole structure. In some embodiments, a segment consists of such discrete parts. In some embodiments, a segment consists of or is contained within characteristic structural elements or parts found in the whole. In some embodiments, the antigen-binding fragment comprises or is composed of at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more monomeric units (e.g., amino acids) found throughout the antibody. In some embodiments, the antigen-binding fragment comprises or is composed of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of monomeric units (e.g., residues) found in the whole antibody. In some embodiments, the nucleotide fragment comprises or is composed of at least about 5%, 10%, 15%, 20%, 25%, 30%, 25%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of monomeric units (e.g., residues) found in the whole nucleotide.

[0078] Frame region: As used herein, the term "frame region" refers to the sequence of the variable region minus the CDR. Because the CDR sequence can be determined using different systems, the frame sequence is subject to correspondingly different interpretations. Six CDRs divide the frame region on both the heavy and light chains into four subregions (FR1, FR2, FR3, and FR4) on each chain, where CDR1 is located between FR1 and FR2, CDR2 between FR2 and FR3, and CDR3 between FR3 and FR4. Unless a specific subregion is designated as FR1, FR2, FR3, or FR4, as otherwise mentioned, a frame region refers to a combination of FRs within the variable region of a single naturally occurring immunoglobulin chain. As used herein, FR represents one of the four subregions; for example, FR1 represents the first frame region closest to the amino terminus of the variable region and located at 5' relative to CDR1, and FR represents two or more subregions constituting the frame region.

[0079] Gene: As used herein, the term "gene" refers to a DNA sequence in a chromosome that encodes a product (e.g., an RNA product and / or a polypeptide product). In some embodiments, a gene contains a coding sequence (i.e., a sequence that encodes a specific product); in some embodiments, a gene contains a non-coding sequence. In some embodiments, a gene may contain both coding sequences (e.g., exons) and non-coding sequences (e.g., introns). In some embodiments, a gene may contain one or more regulatory elements that, for example, can control or influence one or more aspects of gene expression (e.g., cell type-specific expression and / or inducible expression).

[0080] Homology: As used herein, the term "homology" refers to the overall relevance between polymer molecules, such as between nucleic acids (e.g., DNA and / or RNA) and / or between peptides. In some embodiments, polymer molecules are considered "homological" if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, polymer molecules are considered "homological" if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., containing residues with relevant chemical properties at corresponding positions). As those skilled in the art will understand, various algorithms are available that allow comparison of sequences to determine their degree of homology, including allowing vacancies of a specified length in one sequence relative to another when considering which residues "correspond" to each other in different sequences. For example, the percentage of homology between two nucleic acid sequences can be calculated by aligning the two sequences for optimal comparison purposes (e.g., vacancies can be introduced into one or both of the first and second nucleic acid sequences to achieve optimal alignment, and non-corresponding sequences can be ignored for comparison purposes). In some embodiments, the length of the sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of the reference sequence. Nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, the molecules are identical at that position; when a position in the first sequence is occupied by a nucleotide similar to the corresponding position in the second sequence, the molecules are similar at that position. Considering the number of vacancies that need to be introduced for optimal alignment of the two sequences and the length of each vacancy, the percentage of homology between the two sequences is a function of the number of identical and similar positions shared by these sequences.

[0081] Host cell: As used herein, the term "host cell" refers to a cell in which foreign DNA (recombinant or otherwise) has been introduced. Those skilled in the art will understand upon reading this disclosure that such a term refers not only to a specific subject cell but also to its offspring. Because certain modifications may occur in offspring due to mutations or environmental influences, these offspring may actually differ from the parent cell but are still included within the scope of the term "host cell" as used herein. In some embodiments, the host cell includes prokaryotic and eukaryotic cells selected from any living organism suitable for expressing foreign DNA (e.g., recombinant nucleic acid sequences). Exemplary cells include prokaryotic and eukaryotic cells (single-celled or multi-celled), bacterial cells (e.g., strains of *Escherichia coli*, *Bacillus* spp., or *Streptomyces* pp.), mycobacterial cells, fungal cells, yeast cells (e.g., *Saccharomyces cerevisiae*, *Saccharomyces pombe*, *Pichia pastoris*, or *Pichia methanolica*), plant cells, insect cells (e.g., SF-9, SF-21, baculovirus-infected insect cells, or *Trichoplusia ni*), non-human animal cells, human cells, or cell fusions (e.g., hybridomas or quadromas). In some embodiments, the cells include human, monkey, ape, hamster, rat, or mouse cells. In some embodiments, the cells are selected from the following eukaryotic cells: CHO (e.g., CHO Kl, DXB-1 CHO, Veggie-CHO), COS (e.g., COS-7), retinal cells, Vero, CV1, kidney cells (e.g., HEK293, 293 EBNA, MSR 293, MDCK, HaK, BHK), HeLa, HepG2, WI38, MRC 5, Colo205, HB 8065, HL-60 (e.g., BHK21), Jurkat, Daudi, A431 (epidermal), CV-1, U937, 3T3, L cells, C127 cells, SP2 / 0, NS-0, MMT 060562, Sertoli cells, BRL 3A cells, HT1080 cells, myeloma cells, tumor cells, or cell lines derived from the above cells. In some embodiments, the cells contain one or more viral genes.

[0082] Human Antibody: As used herein, the term "human antibody" refers to an antibody having variable and constant regions generated (or assembled) from human immunoglobulin sequences. Antibodies or their antigen-binding fragments may be considered "human" even if their amino acid sequences include residues or elements not encoded by human germline immunoglobulin sequences (e.g., sequence variations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo), such as in one or more CDRs, particularly CDR3.

[0083] Humanization: As used herein, the term "humanization" refers to an antibody or antigen-binding fragment thereof whose amino acid sequence comprises the VH and / or VL region sequences of a reference antibody produced in a non-human species (e.g., mouse), but also comprises modifications to these sequences relative to the reference antibody, modifications intended to make them more "human-like" or more similar to human germline variable region sequences. In some embodiments, the humanized antibody or antigen-binding fragment thereof is an antibody that specifically binds to the antigen of interest and has an FR region having a substantially human antibody amino acid sequence and a CDR having a substantially non-human antibody amino acid sequence. The humanized antibody comprises substantially all at least one and typically two variable domains (Fab, Fab', F(ab')2, FabC, Fv), wherein all or substantially all of the CDR regions correspond to non-human immunoglobulins (e.g., donor immunoglobulins), and all or substantially all of the frame regions correspond to human immunoglobulin common sequences. In some embodiments, the humanized antibody also comprises at least a portion of the immunoglobulin constant region (Fc), typically at least a portion of the human immunoglobulin constant region. In some embodiments, the humanized antibody comprises a light chain and at least a variable domain of the heavy chain. The antibody may also comprise the CH1, hinge, CH2, CH3, and optionally CH4 regions of the heavy chain constant region. In some embodiments, the humanized antibody comprises only the humanized VL region. In some embodiments, the humanized antibody comprises only the humanized VH region. In some embodiments, the humanized antibody comprises both the humanized VH and VL regions.

[0084] Identity: As used herein, the term "identity" refers to the subunit sequence identity between two polymer molecules, particularly between two amino acid molecules such as two polypeptide molecules. Two amino acid sequences are identical when they have the same residues at the same positions; for example, if each of two polypeptide molecules has a position occupied by arginine, then they are identical at that position. The identity or degree of identical residues at the same positions in an alignment of two amino acid sequences is usually expressed as a percentage. The identity between two amino acid sequences is a direct function of the number of matching or identical positions; for example, if half the positions in two sequences (e.g., five positions in a polymer of 10 amino acids in length) are identical, then the two sequences have 50% identity; if 90% of the positions (e.g., nine positions in a polymer of 10 amino acids in length) are identical, then the two amino acid sequences have 90% sequence identity.

[0085] Immune cells: As used herein, the term “immune cell” refers to cells that participate in an immune response (e.g., promote an immune response). Examples of immune cells include, but are not limited to, T cells, natural killer (NK) cells, macrophages, monocytes, dendritic cells, neutrophils, eosinophils, mast cells, platelets, large granular lymphocytes, Langerhans cells, or B lymphocytes.

[0086] Immune checkpoints: As used in this article, the term “immune checkpoint” refers to a group of molecules on the cell surface of CD4+ and / or CD8+ T cells and NK cells that fine-tune the immune response by downregulating or inhibiting the anti-tumor immune response. Immune checkpoint proteins are well known in the art and include, but are not limited to, HHLA2, KIR family receptors, CTLA-4, PD-1, VISTA, B7-H2, B7-H3, PD-L1, B7-H4, B7-H6, ICOS, HVEM, PD-L2, CD160, gp49B, PIR-B, TIM-1, TIM-3, TIM-4, LAG-3, GITR, 4-IBB, OX-40, BTLA, SIRPα, CD47, CD48, 2B4 (CD244), B7.1, B7.2, ILT-2, ILT-4, TIGIT, CD226, CD155, CD112, prolactin, and A2aR. In some embodiments, NK cells contain TIGIT, CD226, and / or CD96. The term also encompasses bioactive protein fragments, as well as nucleic acids encoding full-length immune checkpoint proteins and their bioactive protein fragments. In some implementations, the term also covers any fragments of the homology description provided herein.

[0087] Immune response: As used herein, the term "immune response" refers to a cellular and / or systemic response to an antigen that occurs when lymphocytes identify an antigen molecule as a foreign substance and induce antibody formation and / or activate lymphocytes to remove the antigen. In some embodiments, an immune cellular response may include the proliferation of immune effector cells (e.g., T cells), cytokine production by immune effector cells (e.g., T cells), and / or the release of cytotoxic particles containing perforin and / or granzymes from immune effector cells (e.g., T cells).

[0088] Immunoglobulins: As used herein, the term "immunoglobulin" or "Ig" refers to a class of proteins that act as antibodies. Antibodies expressed by B cells are sometimes referred to as BCRs (B cell receptors) or antigen receptors. The five members of this class of proteins are IgA, IgG, IgM, IgD, and IgE. IgA is a primary antibody found in bodily secretions such as saliva, tears, breast milk, gastrointestinal secretions, and mucus secretions from the respiratory and genitourinary tracts. IgG is the most common circulating antibody. IgM is the major immunoglobulin produced in the primary immune response of most subjects. It is the most effective immunoglobulin in agglutination, complement fixation, and other antibody responses, and is important in the defense against bacteria and viruses. IgD is an immunoglobulin that does not have known antibody functions but can act as an antigen receptor. IgE is an immunoglobulin that mediates immediate-type hypersensitivity reactions by inducing the release of mediators from mast cells and basophils upon exposure to allergens.

[0089] “Improvement,” “Increase,” “Inhibition,” or “Reduction”: As used herein, the terms “improvement,” “increase,” “inhibition,” “reduction,” or their syntactic equivalents, refer to values ​​relative to a baseline or other reference measurement. In some embodiments, an appropriate reference measurement is or includes a measurement in a particular system (e.g., in a single individual) under conditions where the specific agent or treatment is not present (e.g., before and / or after the specific agent or treatment) or in the presence of an appropriate comparable reference agent. In some embodiments, an appropriate reference measurement is or includes a measurement in a comparable system that is known or expected to respond in a particular manner in the presence of a relevant agent or treatment.

[0090] Separation: As used herein, the term "separation" refers to a substance that has been altered or removed from its natural state. For example, nucleic acids or peptides that are naturally present in living organisms are not "separated," but the same nucleic acids or peptides that are partially or completely separated from their natural coexisting substances are "separated." Separated nucleic acids or proteins may exist in substantially purified forms or may exist in non-natural environments such as host cells.

[0091] K D As used in this article, the term "K" D"K" refers to the dissociation constant of a complex of a binder (e.g., an antibody or its antigen-binding fragment) from its partner (e.g., an epitope to which the antibody or its antigen-binding fragment binds). As used herein, the term "K" is... D, "equals K" off Divide by K on .

[0092] K off As used in this article, the term "K" off "" refers to the dissociation rate constant of a conjugate (e.g., an antibody or its antigen-binding fragment) from its complex with its partner (e.g., an epitope to which the antibody or its antigen-binding fragment is bound).

[0093] K on： As used in this article, the term "K" on "" refers to the association rate constant of the association between a binder (e.g., an antibody or its antigen-binding fragment) and its partner (e.g., an epitope bound to an antibody or its antigen-binding fragment).

[0094] Modulation: As used herein, the term “modulation” refers to a detectable increase or decrease in the level and / or nature of a response in a subject compared to the level and / or nature of a response in an untreated or untreated subject. This term encompasses perturbing and / or influencing natural signals or responses, thereby mediating a beneficial therapeutic response in the subject, preferably in a human subject.

[0095] Monoclonal antibodies: "Monoclonal antibody" or "mAb" refers to an antibody derived from a substantially homologous group of antibodies, such that the individual antibodies in that group are identical and / or bind to the same epitope, except for possible variant antibodies (e.g., those containing naturally occurring mutations or generated during the production of the monoclonal antibody), which are typically present in small amounts. Unlike polyclonal antibody formulations, which usually contain different antibodies targeting different determinants (epitaxes), each monoclonal antibody in a monoclonal antibody formulation targets a single determinant on the antigen.

[0096] Operable ligation: As used herein, the term "operable ligation" refers to, for example, a functional ligation between a regulatory sequence and a heterologous nucleic acid sequence that enables the latter to be expressed. For example, the first and second nucleic acid sequences are operably ligated when they are in a functional relationship. For example, if a promoter affects the transcription or expression of a coding sequence, the promoter is operably ligated to the coding sequence. Typically, operably ligated DNA sequences are contiguous and, when ligation of two protein-coding regions is required, are located within the same reading frame.

[0097] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” means compounds, materials, compositions and / or dosage forms that are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, within the bounds of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio.

[0098] Pharmaceutically acceptable carriers: As used herein, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that involves carrying or transporting a subject compound from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of compatibility with other components of the formulation and harmlessness to the subject. Some examples of materials that can be used as pharmaceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; astragalus gum powder; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic and compatible substances used in pharmaceutical formulations.

[0099] Polynucleotide: As used herein, the term "polynucleotide" refers to a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Therefore, nucleic acids and polynucleotides are used interchangeably as used herein. Those skilled in the art will generally know that nucleic acids are polynucleotides that can be hydrolyzed into monomeric "nucleotides." Monomeric nucleotides can be hydrolyzed into nucleosides. As used herein, polynucleotides include, but are not limited to, all nucleic acid sequences obtained by any means available in the art, including but not limited to recombinant methods (i.e., cloning nucleic acid sequences from recombinant libraries or cell genomes using common cloning techniques and PCR™, etc.) and synthetic methods.

[0100] Polypeptide: As used herein, and as may be used interchangeably herein, the term "polypeptide" or "protein" refers to any polymeric chain of residues (e.g., amino acids) typically linked by peptide bonds. In some embodiments, the polypeptide has an amino acid sequence that is naturally occurring. In some embodiments, the polypeptide has an amino acid sequence that is not naturally occurring. In some embodiments, the polypeptide has an engineered amino acid sequence because it is designed and / or generated through artificial action. Polypeptides may comprise natural amino acids, non-natural amino acids, or both, or consist of natural amino acids, non-natural amino acids, or both. Polypeptides may comprise only natural amino acids, only non-natural amino acids, or consist of only natural amino acids, or only non-natural amino acids. Polypeptides may comprise D-amino acids, L-amino acids, or both. Polypeptides may include one or more side groups or other modifications, such as modifications or attachments to one or more amino acid side chains at the N-terminus, C-terminus, or both. In some embodiments, such side groups or modifications are selected from acetylation, amidation, esterification, methylation, or polyethylene glycolation, including combinations thereof. Polypeptides may be cyclic and / or may contain cyclic moieties. In some embodiments, the polypeptide is not cyclic and / or does not contain any cyclic moieties. In some embodiments, the polypeptide is linear. A polypeptide can be or includes pinned polypeptides. The term "polypeptide" may be appended to the name of a reference polypeptide, activity, or structure; in this context, it is used herein to refer to polypeptides that share a relevant activity or structure and are therefore considered members of the same class or family of polypeptides. For each such class, exemplary polypeptides within that class are provided in this specification and / or will be known to those skilled in the art, whose amino acid sequences and / or functions are known; in some embodiments, such exemplary polypeptides are reference polypeptides for a polypeptide class or family. In some embodiments, members of a polypeptide class or family exhibit significant sequence homology or identity with a reference polypeptide of that class, share common sequence motifs (e.g., characteristic sequence elements) with a reference polypeptide of that class, and / or share common activities with a reference polypeptide of that class (at a comparable level or within a specified range in some embodiments). For example, a member polypeptide may have an overall degree of sequence homology or identity with a reference polypeptide, i.e., at least about 30%-40%, and typically about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, and / or include at least one region that exhibits very high sequence identity, typically greater than 90% or even 95%, 96%, 97%, 98% or 99% (e.g., may be or contain a conserved region of characteristic sequence elements).Such conserved regions typically encompass at least 3-4 and usually at most 20 or more amino acids; in some embodiments, the conserved region encompasses at least a segment of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more consecutive amino acids. A useful polypeptide may contain fragments of a parent polypeptide or consist of fragments of a parent polypeptide. A useful polypeptide may contain multiple fragments or consist of multiple fragments, each found in the same parent polypeptide in a spatial arrangement relative to each other that differs from its finding in the polypeptide of interest (e.g., fragments directly linked in the parent may be spatially separated in the polypeptide of interest, and vice versa, and / or fragments may be present in the polypeptide of interest in a different order than in the parent), such that the polypeptide of interest is a derivative of its parent polypeptide.

[0101] Single-chain antibodies: As used herein, the term "single-chain antibody" refers to an antibody formed using recombinant DNA technology, in which segments of the immunoglobulin heavy and light chains are linked to the Fv region via engineered amino acid spans. Various methods for generating single-chain antibodies are known, including those described in the following literature: US Patent No. 4,694,778; Bird (1988) Science 242:423-442; Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883; Ward et al. (1989) Nature 334:54454; Skerra et al. (1988) Science 242:1038-1041.

[0102] Recombinant: As used herein, this term refers to polypeptides designed, engineered, prepared, expressed, produced, manufactured, and / or isolated by recombinant means, such as polypeptides expressed using recombinant expression vectors transfected into host cells; polypeptides isolated from recombinant human polypeptide libraries (see, for example, Hoogenboom, TIB Tech 15:62, 1997; Azzazy Clin. Biochem. 35:425, 2002; Gavilondo BioTechniques 29:128, 2002; Hoogenboom Immunology Today 21:371, 2000); antibodies isolated from animals (e.g., mice) that are transgenic for human immunoglobulin genes (see, for example, Taylor Nuc. Acids Res. 20:6287, 1992; Little Immunology Today 12:364, 2000; Kellermann Curr. Opin. Biotechnol). 13:593,2002; Murphy Proc. Natl Acad Sci USA 111:5153, 2104); or polypeptides prepared, expressed, generated, or isolated by any other means involving the splicing of selected sequence elements with each other. In some embodiments, one or more of such selected sequence elements are found in nature. In some embodiments, one or more of such selected sequence elements are designed by computer simulation. In some embodiments, one or more of such selected sequence elements are generated by mutagenesis (e.g., in vivo or in vitro) of known sequence elements, for example, from natural or synthetic sources. For example, in some embodiments, the recombinant antibody polypeptide consists of sequences present in a germline of the source organism of interest (e.g., human, mouse, etc.). In some embodiments, the recombinant antibody has an amino acid sequence generated by mutagenesis (e.g., in vitro or in vivo, e.g., in transgenic animals) such that the amino acid sequences of the VH and VL regions of the recombinant antibody are sequences that, although derived from and associated with germline VH and VL sequences, are not naturally present in a germline antibody library in vivo.

[0103] Subject: As used herein, the term "subject" refers to an organism, such as a mammal (e.g., a human, a non-human mammal, a non-human primate, a primate, a laboratory animal, a mouse, a rat, a hamster, a gerbil, a cat, or a dog). In some embodiments, the human subject is an adult, adolescent, or pediatric subject. In some embodiments, the subject has a disease, condition, or illness, such as a disease, condition, or illness that can be treated as provided herein, such as cancer or tumors listed herein. In some embodiments, the subject is susceptible to a disease, condition, or illness. In some embodiments, susceptible subjects tend to and / or exhibit an increased risk of developing a disease, condition, or illness (compared to the average risk observed in a reference subject or population). In some embodiments, the subject exhibits one or more symptoms of a disease, condition, or illness. In some embodiments, the subject does not exhibit specific symptoms (e.g., clinical manifestations of a disease) or characteristics of a disease, condition, or illness. In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, condition, or illness. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual who has received and / or has received a diagnostic and / or therapeutic treatment.

[0104] Substantial Identity: As used herein, the term “substantial identity” refers to a comparison between amino acid or nucleic acid sequences. As will be understood by those skilled in the art, two sequences are generally considered “substantially identical” if they contain the same residues at corresponding positions. As is well known in the art, various algorithms, including those available in commercial computer programs such as BLASTN for nucleotide sequences and BLASTP, gap BLAST, and PSI-BLAST for amino acid sequences, can be used to compare amino acid or nucleic acid sequences. In some embodiments, two sequences are considered substantially identical if at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more of the corresponding residues are identical on the relevant residue segment. In some embodiments, the relevant segment is a complete sequence. In some implementations, the relevant segment is at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500 or more residues. In the context of a CDR, reference to "substantially identical" generally refers to a CDR having an amino acid sequence that is at least 80%, preferably at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% identical to the reference CDR.

[0105] Essentially: As used herein, the term “essentially” refers to a qualitative condition that exhibits a general or near-general extent or degree of the characteristic or property of interest. Those skilled in the art of biology will understand that biological and chemical phenomena rarely (if at all) reach completion, and / or proceed to completion or achieve or avoid absolute results. Therefore, the term “essentially” is used herein to capture the inherent lack of completeness in many biological and chemical phenomena.

[0106] "Having": refers to an individual who has been diagnosed with a disease, condition, and / or illness and / or exhibits one or more symptoms of the disease, condition, and / or illness.

[0107] Target: As used herein, the term “target” refers to an agent, cell, tissue, organ or site in the body of a subject who is a recipient of the provided methods, systems, compounds and / or compositions, such as cells, tissues, organs or sites in the body that require treatment or are preferentially bound by, for example, HHLA2 binders described herein.

[0108] Therapeutic: As used herein, the term “therapeutic” refers to treatment and / or prevention. Therapeutic effects are achieved, for example, by suppressing, alleviating, or eradicating a disease state.

[0109] Therapeutic agent: As used herein, the phrase "therapeutic agent" means any agent that, when administered to a subject, has a therapeutic effect and / or induces the desired biological and / or pharmacological action. In some embodiments, a therapeutic agent may be an agent that, when administered to a subject, prevents undesirable side effects. In some embodiments, a therapeutic agent is any substance that can be used to reduce, improve, alleviate, inhibit, prevent, or delay the onset of one or more symptoms or characteristics of a disease, symptom, and / or condition, reduce its severity, and / or decrease its incidence. Therapeutic agents include, but are not limited to, at least one HHLA2 binder as described herein.

[0110] Therapeutic Effective Amount: As used herein, the term "therapeutic effective amount" means the amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that, when administered as part of a treatment regimen, elicits a desired biological response. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, prevent, and / or delay the onset of a disease, condition, and / or symptom when administered to a subject who has or is susceptible to such a disease, condition, and / or symptom. As will be understood by those skilled in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance to be delivered, and / or the target cells or tissues. For example, an effective amount of a compound in a formulation for treating a disease, condition, and / or symptom is an amount that reduces, improves, alleviates, inhibits, prevents, delays, reduces, and / or reduces the incidence of one or more symptoms or features of a disease, condition, and / or symptom. In some embodiments, the therapeutically effective amount is administered in a single dose. In some embodiments, multiple unit doses are required to deliver the therapeutically effective amount.

[0111] Treatment: As used herein, the term "treatment" refers to partial or complete relief, improvement, delayed onset, suppression, prevention, remission, and / or reduction in the incidence and / or severity of one or more symptoms or features of a disease, symptom, and / or condition. In some embodiments, treatment is administered to subjects who do not exhibit signs or features of a disease, symptom, and / or condition (e.g., it may be preventative). In some embodiments, treatment is administered to subjects who exhibit only early or mild signs or features of a disease, symptom, and / or condition, for example, for the purpose of reducing the risk of developing pathology associated with the disease, symptom, and / or condition. In some embodiments, treatment is administered to subjects who exhibit definitive, severe, and / or late signs of a disease, symptom, or condition.

[0112] Tumor: As used herein, the term "tumor" refers to the abnormal growth of cells or tissue. Tumors can contain precancerous (e.g., benign), malignant, pre-metastatic, metastatic, and / or non-metastatic cells. In some embodiments, a tumor is associated with cancer or is a manifestation of cancer. In some embodiments, a tumor is a diffuse or liquid-filled tumor. In some embodiments, a tumor is a solid tumor.

[0113] Throughout this disclosure, various aspects of the provided technology may be presented in range form. It should be understood that the range form is merely for convenience and brevity and should not be construed as a rigid limitation on the scope of the provided technology. Therefore, a range description should be considered to have all possible sub-ranges of the specific disclosure, as well as individual numerical values ​​within that range. For example, a range such as 1 to 6 should be considered to have specific sub-ranges (such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc.) and individual numerical values ​​within that range (e.g., 1, 2, 2.7, 3, 4, 5, 5.3, and 6). This applies regardless of the width of the range.

[0114] As used in this disclosure, unless the context clearly specifies otherwise, (i) the term “a / an” may be understood to mean “at least one / at least one”; (ii) the term “or” may be understood to mean “and / or”; (iii) the terms “comprising”, “comprise”, “including” (whether or not used with “not limited to”) and “include” (whether or not used with “not limited to”) may be understood to cover the listed components or steps, whether presented alone or together with one or more other components or steps; (iv) the term “another” may be understood to mean at least another / second one or more; (v) the terms “about” and “approximately” may be understood to allow standard deviations as understood by one of ordinary skill in the art; and (vi) where a scope is provided, endpoints are included. Detailed Implementation

[0115] In some embodiments, this disclosure provides conjugates. In some embodiments, the conjugate includes a target-binding portion, a payload portion, and a connector connecting the target-binding portion and the payload portion. In some embodiments, this disclosure provides techniques for manufacturing the conjugate. In some embodiments, this disclosure provides techniques for evaluating the conjugate. In some embodiments, this disclosure provides conjugate compositions, such as pharmaceutical compositions. In some embodiments, this disclosure provides methods for using the provided conjugates and compositions thereof. In some embodiments, this disclosure provides techniques for preventing or treating various conditions, symptoms, or diseases using the provided conjugates and compositions thereof.

[0116] The following describes some implementation schemes (including those for target bonding portions, payload portions, connectors, conjugates, etc.) as examples.

[0117] Target binding part The target-binding portion binds to a target, which may be or include peptides, small molecules, carbohydrates, lipids, or nucleic acids. In some embodiments, the target is on the surface of cells (e.g., cancer cells).

[0118] In some embodiments, the target binding moiety is or comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the target binding moiety is an antibody or an antigen-binding fragment thereof. In some embodiments, the target binding moiety is an antibody. In some embodiments, the target binding moiety is a monoclonal antibody. In some embodiments, the target binding moiety is not trastuzumab. In some embodiments, the target binding moiety shares less than about 90%, 95%, or 99% sequence identity with trastuzumab. In some embodiments described herein, the target binding moiety shares less than about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with trastuzumab. In some embodiments, it is less than about 95%. In some embodiments, it is less than about 90%. In some embodiments, it is less than about 85%. In some embodiments, the target binding moiety is not an anti-HER2 antibody. In some embodiments, the target binding moiety is not an anti-HER2 antibody or an antigen-binding fragment thereof.

[0119] In some embodiments, the target binding portion is an antibody portion containing four Cys residues forming interchain disulfide bonds between the two heavy chains and the light chain without conjugation. In some embodiments, each of the four Cys residues is independently conjugated to the payload portion. In some embodiments, the target binding portion is an antibody portion containing eight residues forming interchain disulfide bonds without conjugation. In some embodiments, each of the eight Cys residues is independently conjugated to the payload portion.

[0120] Those skilled in the art will understand that, according to this disclosure, various antibodies (e.g., those approved by the FDA, those that can bind to the same antigen or entity (e.g., peptides, cells, etc.)) and their antigen-binding fragments can be used as target binding moieties. For example, in some embodiments, the target binding moieties are anti-HHLA2 antibodies or their antigen-binding fragments.

[0121] In some embodiments, the target binding portion is or comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the target binding portion comprises an antibody. In some embodiments, the target binding portion is an antibody. In some embodiments, the target binding portion comprises an antigen-binding fragment of an antibody. In some embodiments, the target binding portion is an antigen-binding fragment of an antibody. In some embodiments, the antibody is a chimeric antibody.

[0122] In some embodiments, the antibody is a human antibody. In some embodiments, the antibody is a humanized antibody. In some embodiments, the antibody is a monospecific antibody. In some embodiments, the antibody is a bispecific antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the antibody or its antigen-binding fragment is or contains a heavy chain constant region or a fragment thereof. In some embodiments, the heavy chain constant region is IgG1. In some embodiments, the heavy chain constant region is IgG2. In some embodiments, the heavy chain constant region is IgG3. In some embodiments, the heavy chain constant region is IgG4. Alternatively or additionally, the antibody or its antigen-binding fragment is or contains a light chain constant region. In some embodiments, the light chain constant region is the κ light chain constant region. In some embodiments, the light chain constant region is the λ light chain constant region.

[0123] In some embodiments, the target binding moiety is or comprises a chimeric antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is independently or comprises a human antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is independently or comprises a humanized antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is independently or comprises a monospecific antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is independently or comprises a bispecific antibody or its antigen-binding fragment (e.g., an HHLA2 binding arm and at least one other non-HHLA2 binding arm). In some embodiments, the target binding moiety is independently or comprises an anti-EGFR×HER3 bispecific antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is independently or comprises a double complementary site antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is an anti-HHLA2 antibody or its antigen-binding fragment. In some embodiments, the antibody or its antigen-binding fragment is an anti-HER2×HHLA2 bispecific antibody or its antigen-binding fragment. In some embodiments, such an antibody or its antigen-binding fragment comprises the CDR sequence of trastuzumab. In some implementations, such antibodies or their antigen-binding fragments contain the antigen-binding sequence of trastuzumab.

[0124] For example, in some embodiments, the target binding portion is or contains an anti-HHLA2 antibody or its antigen-binding fragment.

[0125] In some embodiments, the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) Heavy chain variable regions (VHs) comprising one, two, or three VH CDR sequences each having at least approximately 90% identity with the VH CDRs in Table 1; and / or (b) Light chain variable region (VL) which contains one, two or three VL CDR sequences that each have at least about 90% identity with the VL CDRs in Table 1.

[0126] In some embodiments, the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) VH, comprising one, two, or three VH CDR sequences each having at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH CDRs in Table 1; and / or (b) VL, which comprises one, two or three VL CDR sequences each having at least about 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 99.5% identity with the VL CDRs of Table 1.

[0127] In some embodiments, the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) VH, which contains one, two or three sequences of VH CDRs, each containing or composed of VH CDRs listed in Table 1; and / or (b) VL, which contains one, two or three VL CDRs, each containing or consisting of a VL CDR from Table 1.

[0128] In some embodiments, the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) VH that has at least about 90% or more identity with the VH in Table 1; and / or (b) VL, which has at least about 90% or more identity with the VL in Table 1.

[0129] In some embodiments, the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) VH that has at least about 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH in Table 1; and / or (b) VL, which has at least about 95%, 96%, 97%, 98%, 99% or 99.5% identity with the VL in Table 1.

[0130] In some implementations, anti-HHLA2 or its antigen-binding fragment is or comprises: (a) VH, which includes or consists of the VH in Table 1; and / or (b) VL, which includes or consists of the VL in Table 1.

[0131] In some implementations, anti-HHLA2 or its antigen-binding fragment is or comprises: (a) Heavy chains that have at least about 90% or more identity with the heavy chains in Table 1; and / or (b) Light chains that have at least about 90% or more of the same identity as the light chains in Table 1.

[0132] In some implementations, anti-HHLA2 or its antigen-binding fragment is or comprises: (a) Heavy chains that have at least about 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the heavy chains in Table 1; and / or (b) Light chains that have at least about 95%, 96%, 97%, 98%, 99% or 99.5% identity with the light chains in Table 1.

[0133] In some implementations, anti-HHLA2 or its antigen-binding fragment is or comprises: (a) Heavy chains that contain or consist of the heavy chains in Table 1; and / or (b) Light chains, which contain or consist of the light chains in Table 1.

[0134] In some embodiments, the target binding portion comprises each of the CDRs of 887-1DE in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH of 887-1DE in Table 1. In some embodiments, the target binding portion is or comprises a VH of 887-1DE in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VL of 887-1DE in Table 1. In some embodiments, the target binding portion is or comprises a VL of 887-1DE in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the heavy chain of 887-1DE in Table 1. In some embodiments, the target binding moiety is or comprises the heavy chain of 887-1DE in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the light chain of 887-1DE in Table 1. In some embodiments, the target binding moiety comprises 887-1DE or its antigen-binding fragment in Table 1. In some embodiments, the target binding moiety is 887-1DE in Table 1.

[0135] In some embodiments, the target binding portion comprises each of the CDRs of 65889 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH of 65889 in Table 1. In some embodiments, the target binding portion is or comprises a VH of 65889 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VL of 65889 in Table 1. In some embodiments, the target binding portion is or comprises the VL of 65889 in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the heavy chain of 65889 in Table 1. In some embodiments, the target binding moiety is or comprises the heavy chain of 65889 in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the light chain of 65889 in Table 1. In some embodiments, the target binding moiety comprises 65889 in Table 1 or an antigen-binding fragment thereof. In some embodiments, the target binding moiety is 65889 in Table 1.

[0136] In some embodiments, the target binding portion comprises each of the CDRs of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a VH of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VL of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a VL of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the heavy chain of 65872 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the light chain of 65872 in Table 1. In some embodiments, the target binding portion is or comprises the light chain of 65872 in Table 1. In some embodiments, the target binding portion comprises 65872 in Table 1 or an antigen-binding fragment thereof. In some embodiments, the target binding portion is 65872 in Table 1.

[0137] In some embodiments, the target binding portion comprises each of the CDRs of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VH of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding portion is or comprises a VH of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding portion is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the VL of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding portion is or comprises a VL of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the heavy chain of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding moiety is or comprises the heavy chain of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding moiety is or comprises a sequence having about or at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% identity with the light chain of Hu2C4-H3L3 in Table 1. In some embodiments, the target binding moiety comprises Hu2C4-H3L3 in Table 1 or an antigen-binding fragment thereof. In some embodiments, the target binding moiety is Hu2C4-H3L3 in Table 1.

[0138] In some embodiments, the conjugate or target-binding portion is at a concentration of about or no more than about 1 pM, 5 pM, 10 pM, 15 pM, 20 pM, 30 pM, 50 pM, 100 pM, or 500 pM, or about or no more than about 1 nM, 5 nM, 10 nM, 15 nM, 20 nM, 30 nM, 50 nM, 100 nM, or 500 nM. D It binds to its target, such as HHLA2. In some embodiments, the conjugate or target-binding portion binds to its target, such as HHLA2, with an affinity of about or at least about 10, 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 times greater than the background or reference agent (e.g., no conjugate, immunoglobulin targeting a different target, conjugate targeting a different target, etc.).

[0139] In some embodiments, the conjugate comprises two or more target-binding moieties. In some embodiments, each target-binding moiety of the provided conjugate targets the same agent. In some embodiments, each target-binding moiety targets the same agent, wherein the agent is or comprises a peptide, nucleic acid, or lipid. In some embodiments, each target-binding moiety targets the same agent, wherein the agent is a peptide. In some embodiments, each target-binding moiety targets the same antigen. In some embodiments, each target-binding moiety is independently an antibody or an antigen-binding fragment thereof, and each target-binding moiety shares about or at least about 90%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, each target-binding moiety is independently an antibody or an antigen-binding fragment thereof, and each target-binding moiety shares the same amino acid sequence. In some embodiments, each target-binding moiety is identical. In some embodiments, each target-binding moiety is independently the target-binding moieties described herein.

[0140] Payload portion Various payload portions can be utilized according to this disclosure, including those reported for antibody-drug conjugates. In some embodiments, the payload portion is a portion that is or contains a cytotoxic agent.

[0141] In some embodiments, the active ingredient is or contains calicheamicin or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof. In some embodiments, the active ingredient is or contains N-acetylγ-calicheamicin.

[0142] In some embodiments, the active ingredient is or comprises a topoisomerase inhibitor. In some embodiments, the active ingredient is or comprises camptothecin or its derivatives or analogs, or a pharmaceutically acceptable salt thereof. In some embodiments, the active ingredient is or comprises essanotecan or its derivatives or analogs, or a pharmaceutically acceptable salt thereof.

[0143] Exatecan ((1S,9S)-1-amino-9-ethyl-5-fluoro-9-hydroxy-4-methyl-1,2,3,9,12,15-hexahydro-10H,13H-benzo[de]pyrano[3′,4′:6,7]indolazino[1,2- b Quinoline-10,13-dione is a structural analogue of camptothecin. Exanotecan contains the following chemical structure: .

[0144] This disclosure provides, in particular, various ethatecan conjugates, such as the various conjugates described in the examples. As shown herein, in various embodiments, ethatecan is conjugated to a target-binding moiety (e.g., an antibody) via its -NH2 group through a linker.

[0145] Derivatives of ixanotecan are used in trastuzumab deruxtecan (sold under the brand name Enhertu), which is an antibody-drug conjugate composed of the humanized monoclonal antibody trastuzumab (Herceptin) covalently linked to the topoisomerase I inhibitor deruxtecan (a derivative of ixanotecan). Trastuzumab binds to epidermal growth factor receptor 2 (HER2 / neu), and once bound to the HER2 receptor, the antibody is internalized into the cell, thereby carrying the bound deruxtecan. Trastuzumab deruxtecan has the following chemical structure: .

[0146] Not wanting to be bound by a specific theory, the conjugate binds to its target (e.g., HHLA2), and once bound, the conjugate can be internalized into the cell, thereby carrying the binding payload (e.g., ethanotecan).

[0147] In some implementations, the payload portion is Or its pharmaceutically acceptable salt.

[0148] In some implementations, the payload portion is In some implementations, the payload portion is... .

[0149] In some implementations, the topoisomerase inhibitor is or contains SN-38 or a pharmaceutically acceptable salt thereof.

[0150] In some embodiments, the payload is or comprises a microtubule disruptor. In some embodiments, the payload is or comprises an antimitotic agent. In some embodiments, the payload is or comprises a microtubule inhibitor. For example, in some embodiments, the payload is or comprises MMAE or its derivatives or analogs, or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or comprises MMAE or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or comprises MMAF or its derivatives or analogs, or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or comprises MMAF or a pharmaceutically acceptable salt thereof.

[0151] In some embodiments, the active loading is or contains an alkylating agent. In some embodiments, the active loading is or contains a DNA alkylating agent. In some embodiments, the active loading is or contains pyrrolobenzodiazepine. In some embodiments, the active loading is or contains a pyrrolobenzodiazepine dimer. In some embodiments, the active loading is or contains SG3199 or its derivatives or analogs, or a pharmaceutically acceptable salt thereof. In some embodiments, the active loading is or contains SG3199 or a pharmaceutically acceptable salt thereof.

[0152] In some embodiments, the payload is or comprises a microtubule inhibitor (e.g., maytansine) or a derivative or analogue thereof (e.g., DM1, DM4, etc.), or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or comprises DM1 or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or comprises DM4 or a pharmaceutically acceptable salt thereof.

[0153] In some embodiments, the active ingredient is or comprises auristatin, maytansinoid, duocarmycin, hemiasterlin, eribulin, N-myristyltransferase inhibitor, amanitin, or a derivative or analogue thereof. In some embodiments, the active ingredient is or comprises auristatin or a derivative or analogue thereof. In some embodiments, the active ingredient is or comprises maytansinoid, or a derivative or analogue thereof. In some embodiments, the active ingredient is or comprises hemiasterlin, or a derivative or analogue thereof. In some embodiments, the active ingredient is or comprises eribulin, or a derivative or analogue thereof. In some embodiments, the active ingredient is or comprises N-myristyltransferase inhibitor, or a derivative or analogue thereof (see, for example, WO 2022058745). In some embodiments, the payload is or contains an anthracycline drug or its derivatives or analogs (see, for example, US11833120). In some embodiments, the payload is or contains nemorubicin or its metabolite (e.g., PNU-159682), its derivatives or analogs, or a pharmaceutically acceptable salt thereof. In some embodiments, the payload is or contains PNU-159682, its derivatives or analogs, or a pharmaceutically acceptable salt thereof. See, for example, Holte D et al., Evaluation of PNU-159682 antibody drugconjugates (ADC). Bioorg Med Chem Lett. 15 Dec 2020;30(24):127640. doi:10.1016 / j.bmcl.2020.127640. E-published 28 Oct 2020. PMID: 33127540. In some embodiments, the active ingredient is or comprises PNU-159682 or a pharmaceutically acceptable salt thereof. In some embodiments, the active ingredient is or comprises muscarine or its derivatives or analogs. For example, in some embodiments, the active ingredient portion is or comprises... In some implementations, the payload portion is or includes In some implementations, the payload portion is or includes In some implementations, the payload portion is or includes In some implementations, the payload portion is or includes In some implementations, the payload portion is or includes In some implementations, the payload portion is or includes .

[0154] In some implementations, the payload is or contains a chemotherapeutic agent.

[0155] Conjugates are particularly capable of delivering a payload (e.g., one or more payloads). In some embodiments, the conjugate delivers at least one cytotoxic agent (e.g., two, three, four, or more). In some embodiments, the conjugate delivers at least one chemotherapeutic agent (e.g., two, three, four, or more). In some embodiments, the conjugate delivers camptothecin, aureatin, maytansine alkaloids, pyruvic acid, hammetrine, eribulin, or an N-myristyltransferase inhibitor, or a derivative or analog thereof, or a pharmaceutically acceptable salt thereof.

[0156] The payload can be conjugated at various positions and is typically conjugated at functional groups such as amino or hydroxyl groups. In some embodiments, the functional group is bonded to a secondary or tertiary carbon. In some embodiments, the functional group is bonded to a ring atom. In some embodiments, the functional group is bonded to a cyclic carbon atom. In some embodiments, the ring is a polycyclic system, such as those found in ethathecan or its derivatives or analogues. In some embodiments, the provided technology provides conjugates with functional groups, such as those bonded to secondary or tertiary carbon atoms or cyclic atoms (e.g., cyclic carbon atoms) without spacer moieties. In some embodiments, L C1 It is bonded to the functional group (e.g., amino group) of the load portion at its -C(O)- position. In some embodiments, the load is bonded to the linker at the amino group. In some embodiments, the H-load portion is the corresponding load for the load portion. In some embodiments, the load portion is the R- as described herein. P .

[0157] In some embodiments, the payload is a small molecule. For example, in some embodiments, the molecular weight is about or no more than about 5000 Da, 4000 Da, 3000 Da, 2500 Da, 2000 Da, 1500 Da, 1000 Da, 900 Da, 800 Da, 700 Da, 600 Da, or 500 Da.

[0158] In some embodiments, in the conjugate, one or more payload moieties (e.g., ethatecan or its derivatives or analogs, or pharmaceutically acceptable salts thereof) are covalently bound to a single target-binding moiety (e.g., an anti-HERV-H LTR-association 2 (HHLA2) antibody or its antigen-binding fragment). In some embodiments, the ratio of payload moieties to target-binding moieties (which is equal to the number of payload moieties (e.g., ethatecan or its derivatives, or pharmaceutically acceptable salts thereof) covalently bound to a single target-binding moiety in various conjugates) is referred to as the drug-to-antibody ratio (DAR). In some embodiments, the provided conjugates have a DAR between about 1 and about 20. In some embodiments, the provided conjugates have a DAR between about 1 and about 16. In some embodiments, the provided conjugates have a DAR between about 1 and about 12. In some embodiments, the provided conjugates have a DAR between about 2 and about 15. In some embodiments, the provided conjugates have a DAR between about 2 and about 12. In some embodiments, the provided conjugates have a DAR between about 2 and about 10. In some embodiments, the provided conjugate has a DAR between about 2 and about 8. In some embodiments, the provided conjugate has a DAR between about 4 and about 8. In some embodiments, the provided conjugate has a DAR between about 7 and about 9. In some embodiments, the provided conjugate has a DAR between about 6 and about 10. In some embodiments, the provided conjugate has a DAR between about 4 and about 12. In some embodiments, the provided conjugate has a DAR of about 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, the provided conjugate has a DAR of about 2, 4, 6, or 8. In some embodiments, the provided conjugate has a DAR of about 2 or greater. In some embodiments, the provided conjugate has a DAR of about 2. In some embodiments, the provided conjugate has a DAR of about 4. In some embodiments, the provided conjugate has a DAR of about 8. In some embodiments, the payload portion is identical.

[0159] connector In some embodiments, the conjugate comprises one or more linkers, each linker connecting an effective load portion (which may be in the form of a pharmaceutically acceptable salt) to a target binding portion (which may also be in the form of a pharmaceutically acceptable salt). In some embodiments, the linker is divalent and connects a single effective load portion to a single target binding portion. In some embodiments, the linker is multivalent and connects one or more effective load portions to one or more target binding portions. In some embodiments, the linker is multivalent and connects one or more effective load portions to a single target binding portion. In some embodiments, the linker is multivalent and connects a single target binding portion to a single effective load portion.

[0160] In some embodiments, the connector comprises -(CH2CH2O)n- as described herein. In some embodiments, the connector comprises L as described herein. C In some implementations, the connector comprises -(CH2CH2O)n- and L as described herein. C In some embodiments, the connector comprises a PEG portion. In some embodiments, the connector comprises -(CH2CH2O)n-, where n is at least 1. In some embodiments, the connector comprises -(CH2CH2O)n-, where n is 1-24. In some embodiments, n is 4. In some embodiments, n is 8.

[0161] In some implementations, the connector contains -(CH2CH2O)nL ZP -L C - where each variable is independent as described in this paper.

[0162] In some implementations, the connector includes -L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described in this paper.

[0163] In some implementations, the connector includes -L RX -L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described in this paper.

[0164] In some implementations, the connector includes -L T -L RX -L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described in this paper.

[0165] In some implementations, the connector is or includes Each variable is independent as described in this paper.

[0166] L ZP As described herein, in some implementations, L ZP Connect -(CH2CH2O)n- to L C Where n is not 0. In some implementations, L ZP Connect L X With L C In some implementations, L ZP There is no (covalent bond). Various moieties, including those used in antibody-drug conjugates, can be used as L according to this disclosure. ZP .

[0167] In some implementations, L ZP It is a covalent bond. In some implementations, L ZP It is a bivalent straight chain or branched chain that can be arbitrarily replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L ZP The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L ZP It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 Aliphatic, etc., of which L ZPOne to five (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L ZP It is a divalent C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) aliphatic. In some implementations, L ZP Includes L C Bonded -C(O)-. In some implementations, L ZP It is -(CH2) Z -C(O)-, where z is 1-19, and each of -CH2- is optionally substituted independently. In some embodiments, L ZP It is -(CH2) Z -C(O)-, where z is 1-19, each of -CH2- is independently and optionally substituted, and L C1 It is or contains a peptide moiety (e.g., GGFG, Val-Ala, Val-Cit, etc.). In some embodiments, L ZP It is -(CH2) Z -C(O)-. In some implementations, L ZP It is -(CH2) Z -N(R')C(O)-, where z is 1-19, each of -CH2- is optionally substituted independently, and R' is as described herein. In some embodiments, L ZP It is -(CH2) Z -N(R')C(O)-, where z is 1-19, each of -CH2- is independently and optionally substituted, R' is as described herein, and L C1 It is L C1S (e.g., oGlu, mGlu, etc.). In some implementations, L ZP It is -(CH2) Z -N(R')C(O)-, where z is 1-19, and R' is as described herein. In some implementations, L ZP It is -(CH2) Z-NHC(O)-, where z is 1-19. In some embodiments, z is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 1-15, 1-10, etc. In some embodiments, z is 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.). In some embodiments, z is 1-5 (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.). In some embodiments, z is 1. In some embodiments, z is 2. In some embodiments, z is 3. In some embodiments, z is 4. In some embodiments, z is 5. In some embodiments, L ZP It is -(CH2)2-C(O)-, where -C(O)- and L C The amino bond of the first amino acid in L. In some implementations, L ZP It is -(CH2)2-C(O)-, where -C(O)- and L C The amino bond of the first amino acid in, where L C Yes -L C1 -L CO - In some implementations, L ZP It is -(CH2)2-N(R')-C(O)-, where -C(O)- and L C1 In the bonding process, each of the -CH2- groups is independently and optionally substituted, R' as described herein, and L C1 It is L C1S In some implementations, L ZP It is -(CH2)2-N(R')-C(O)-, where -C(O)- and L C In the bonding process, each of the -CH2- groups is independently and optionally substituted, R' as described herein, and L C1 It is L C1S (e.g., oGlu, mGlu, etc.). In some implementations, L ZP It is -(CH2)2-NH-C(O)-, where -C(O)- reacts with L C1 Bonding, and L C1 It is L C1S In some implementations, L ZP It is -(CH2)2-NH-C(O)-, where -C(O)- reacts with L C Bonding, and L C1 It is L C1S (e.g., oGlu, mGlu, etc.). For example, as shown in the embodiments, in some implementations, L ZP It is -(CH2)2-NH-C(O)-, and LC It is mGlu; in some implementations, L ZP It is -(CH2)2-NH-C(O)-, and L C It is oGlu. In some implementations, L ZP It is -(CH2)2-C(O)-N(R')-, where -N(R')- and L C1 In the bonding process, each of the -CH2- groups is independently and optionally substituted, R' as described herein, and L C1 It is L C1S In some implementations, L ZP It is -(CH2)2-C(O)-N(R')-, where -N(R')- and L C In the bonding process, each of the -CH2- groups is independently and optionally substituted, R' as described herein, and L C1 It is L C1S (e.g., oGlu, mGlu, etc.). In some implementations, L ZP It is -(CH2)2-C(O)-NH-, where -NH- and L C1 Bonding, and L C1 It is L C1S In some implementations, L ZP It is -(CH2)2-C(O)-NH, where -NH- and L C Bonding, and L C1 It is L C1S (e.g., oGlu, mGlu, etc.). In some implementations, L ZP It is -(CH2)2-C(O)-NH-, and L C It is mGlu; in some implementations, L ZP It is -(CH2)2-C(O)-NH-, and L C It is oGlu.

[0168] L C1 In some implementations, each L C1 Independently, it is a pyrolytic unit as described herein. Variable pyrolytic units may be utilized according to this disclosure. In some embodiments, certain pyrolytic units, when utilized as described herein, may provide various advantages, such as improved properties (e.g., solubility, stability, etc.), activity, etc. In some embodiments, L C1 It may contain a polypeptide. In some embodiments, each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S-Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S - where each variable is independent as described herein. In some implementations, L C1 Is it -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or In some implementations, L C1 It is -Gly-Gly-Phe-Gly-. In some implementations, L C1 It is -Val-Cit-. In some implementations, L C1 It is -Val-Ala. In some implementations, L C1 It is as described in this article, L C1S In some implementations, L C1 It is -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S - In some implementations, L C1 It is -Gly-Gly-Phe-Gly-NH-L C1S -、-Val-Ala-NH-L C1S -or-Val-Cit-NH-L C1S - In some implementations, L C1 yes In some implementations, L C1 yes In some implementations, L C1 yes In some implementations, L C1 yes .

[0169] L C1S and R LS As used in this article, L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component. In some implementations, L C1S yes The carbonyl group and L CO Bonding, and RLS It is or contains a sugar moiety (the benzene ring and -CH2- are unsubstituted). In some embodiments, R LS It is or contains a sugar portion, making L C It is cleavable, for example, by an enzyme. In some embodiments, R LS It may contain a glucuronide moiety. In some embodiments, L C It can be cleaved by β-glucuronidase. In some implementations, R LS It may contain a galactoside moiety. In some embodiments, L C It can be cleaved by β-galactosidase. In some implementations, R LS Yes or include Or its derivatives. In some embodiments, R LS yes In some implementations, R LS Yes or include Or its derivatives. In some embodiments, R LS yes In some embodiments, the sugar moiety is protected, for example, in compounds that can be used to prepare the conjugate. In some embodiments, one or more or all of the -OH groups are protected as, for example, -OC(O)R, where R is as described herein. In some embodiments, R is -CH3. In some embodiments, -COOH is protected as -C(O)OR, where R is as described herein and is not -H. In some embodiments, R is an optionally substituted C 1-6 Alkyl group. In some embodiments, each -OH group is protected as -OAc, and the -COOH group is protected as -C(O)OMe. In some embodiments, R LS yes .

[0170] For example, in some implementations, L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding. In some implementations, L C1S yes The carbonyl group and L CO Or the payload portion (“oGlu”) is bonded. In some implementations, L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding. In some implementations, L C1S yes The carbonyl group and L CO Or the payload portion (“mGlu”) is bonded. In some implementations, LC Is this type of L C1S As will be readily understood by those skilled in the art, various acidic and basic groups may be present in one or more forms. In some embodiments, the group (e.g., -COOH) is in its acidic form. In some embodiments, the group (e.g., -COOH) is in its salt form. In some embodiments, the group (e.g., -COOH) is in a pharmaceutically acceptable salt form. In some embodiments, in a formulation or composition, the group (e.g., -COOH) may be present in multiple forms, such as its acidic form and / or one or more pharmaceutically acceptable salt forms.

[0171] In some implementations, L C1 It is as described in this article, L C1S In some implementations, L CO It is a covalent bond. In some implementations, L C It is as described in this article, L C1 In some implementations, L C It is as described in this article, L C1S .

[0172] L CO In some implementations, L CO It is a covalent bond. In some implementations, L CO It is the spacer sub-part as described herein. In some implementations, L CO It is a spacer. In some implementations, L CO It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L COThe 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L CO It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 Aliphatic, etc., of which L CO One to five (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. For example, in some embodiments, L CO It was arbitrarily replaced. In this configuration, the carbonyl group is bonded to the effective loading portion, and the amino group is bonded to the L group. C1 -C(O)-bonding. In some implementations, L CO yes (PABA). In some implementations, L CO It is a covalent bond, and the payload portion is related to L. C1 -C(O)-bonding. In some implementations, L ZP It is a divalent C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) aliphatic.

[0173] L C As described in this article, L C Partially bonded to the payload and is -L C1 -L CO -, where L C1 and L CO Each of these is independent as described herein. In some implementations, L C It is pyrolytic. In some implementations, L C It is L C1 In some implementations, L C It is LC1S For example, in some implementations, L C It is oGlu. In some implementations, L C It is mGlu. In some implementations, L C Yes -L C1 -L CO -, where L C1 It is Val-Ala, and L CO It was arbitrarily replaced. In some implementations, L C Yes -L C1 -L CO -, where L C1 It is Val-Cit, and L CO It was arbitrarily replaced. In some implementations, L C It is -Val-Ala-NH-L C1S L C1S As described herein. In some implementations, L C It is -Val-Ala-NH-L C1S L C1S As described herein (e.g., mGlu, oGlu, etc.). In some implementations, L C It is -Val-Ala-NH-L C1S -mGlu-. In some implementations, L C It is -Val-Ala-NH-L C1S -oGlu-. In some implementations, L C It is -Val-Cit-NH-L C1S L C1S As described herein (e.g., mGlu, oGlu, etc.). In some implementations, L C It is -Val-Cit-NH-L C1S -mGlu-. In some implementations, L C It is -Val-Cit-NH-L C1S -oGlu-. In some implementations, L C It is -GGFG-NH-L C1S L C1S As described herein (e.g., mGlu, oGlu, etc.). In some implementations, L C It is -GGFG-NH-L C1S -mGlu-. In some implementations, L C It is -GGFG-NH-L C1S -oGlu-.

[0174] In some implementations, L C yes The carbonyl group is bonded to the loaded moiety. In some embodiments, L C yes The carbonyl group is bonded to the loaded moiety. In some embodiments, L C yes The carbonyl group is bonded to the loaded moiety. In some embodiments, L C yes (Val-Cit-PABA), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C yes (oGlc), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C yes (oGlc-a), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C yes (mGlc), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C yes (mGlc-a), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C yes (Val-Ala-PABA), wherein the carbonyl group is bonded to the supported moiety. In some embodiments, L C It is Val-Cit-PABA, wherein the carbonyl group of PABA is bonded to the supported moiety. In some embodiments, L C It is GGFG-PABA, wherein the carbonyl group of PABA is bonded to the supported moiety. In some embodiments, L C It is Val-Ala-NH-oGlu, wherein the carbonyl group of oGlu is bonded to the loaded moiety. In some embodiments, L C It is Val-Cit-NH-oGlu, wherein the carbonyl group of oGlu is bonded to the loaded moiety. In some embodiments, L C It is Val-Ala-NH-mGlu, wherein the carbonyl group of mGlu is bonded to the loaded moiety. In some embodiments, L C It is Val-Cit-NH-mGlu, in which the carbonyl group of mGlu is bonded to the effective loading portion.

[0175] Unless otherwise stated, the polypeptide sequence is from N to C.

[0176] In some implementations, -L ZP -L C- is -(CH2)2C(O)-GGFG-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-Val-Ala-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-Val-Cit-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-GGFG-PABA-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L ZP -L C - is -(CH2)2NHC(O)oGlc-. In some implementations, -L ZP -L C - is -(CH2)2NHC(O)-mGlc-. In some implementations, -L ZP -L C - is -(CH2)2C(O)-(oGlc-a)-. In some implementations, -L ZP -L C -is-(CH2)2C(O)-(mGlc-a)-. -L ZP -L C - is -(CH2)2NHC(O)-mGlu-. In some implementations, -L ZP -L C - is -(CH2)2NHC(O)-oGlu-. In some implementations, -L ZP -L C - It is bonded to the payload portion at its -C(O)-.

[0177] n In some embodiments, the connector comprises -(CH2CH2O)n-, where n is 0-24. In some embodiments, n is 1-24. In some embodiments, n is 6-24. In some embodiments, n is 7-20. In some embodiments, n is 8-15. In some embodiments, n is 5-10. In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. In some embodiments, n is 2. In some embodiments, n is 4. In some embodiments, n is 5, 6, 7, 8, 9, or 10. In some embodiments, n is 8. In some embodiments, n is 10. In some embodiments, n is 0.

[0178] In some implementations, n is 0, and L C It contains a carbohydrate portion. In some implementations, n is 0, and L C It contains a glucuronic acid moiety. In some embodiments, n is 0, and L C It contains the galactose moiety. In some implementations, n is 0, and L C It is oGlc, oGlc-a, mGlc, or mGlc-a. In some implementations, n is 0, and L C It is mGlu. In some implementations, n is 0, and L C It is oGlu.

[0179] In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some embodiments, -(CH2CH2O)nL ZP -L C- is -(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some embodiments, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -(CH2CH2O)nL ZP -L C -is-(CH2CH2O)8-(CH2)2NHC(O)-L C1S - In some implementations, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2NHC(O)-mGlu-. In some embodiments, -(CH2CH2O)nL ZP -L C - is -(CH2CH2O)8-(CH2)2NHC(O)-oGlu-. In some embodiments, -(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-.

[0180] L X In some implementations, L X It is a covalent bond. In some implementations, L X It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L X The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 Aliphatic, etc., of which L X One to five (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X It is -(CH2) X -C(O)-N(R')-. In some implementations, L X It is -(CH2) X -C(O)-N(R')-, where x is 1-19, and -N(R')- is bonded to -(CH2CH2O)n-. In some implementations, L X It is -(CH2) X -C(O)-NH-, where x is 1-19, and -NH- is bonded to -(CH2CH2O)n-. In some embodiments, L X It is -(CH2) X -C(O)-N(R')- or -(CH2) X -C(O)-NH-, and x is 1-6. In some implementations, L XIt is -(CH2)5-C(O)-N(R')-, where -N(R')- is bonded to -(CH2CH2O)n-. In some embodiments, L X It is -(CH2)5-C(O)-NH-, where -NH- is bonded to -(CH2CH2O)n-. In some embodiments, L X It is -(CH2)3-C(O)-N(R')-, where -N(R')- is bonded to -(CH2CH2O)n-. In some embodiments, L X It is -(CH2)3-C(O)-NH-, wherein -NH- is bonded to -(CH2CH2O)n-. In some embodiments, x is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 1-15, 1-10, etc. In some embodiments, x is 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.). In some embodiments, x is 1-5 (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.). In some embodiments, x is 1. In some embodiments, x is 2. In some embodiments, x is 3. In some embodiments, x is 4. In some embodiments, x is 5.

[0181] In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP-L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlu-. In some embodiments, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlu-. In some embodiments, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some embodiments, -L X -(CH2CH2O)nLZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -LX -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - is -(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L X -(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-.

[0182] L RX In some implementations, L X It is a covalent bond. In some implementations, L X It is a divalent or multivalent straight or branched chain C that can be arbitrarily replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L X The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X It is a covalent bond or an optionally substituted divalent or multivalent straight or branched chain C. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10Aliphatic, etc., of which L X One to five methylene units (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, at least one methylene unit is replaced by -Cy-. In some embodiments, -Cy- is... In some implementations, -Cy- is In some implementations, -Cy- is In some implementations, -Cy- is In some embodiments, at least one methylene unit is replaced by -S-. In some embodiments, at least one methylene unit is replaced by -C(O)-.

[0183] In some implementations, each L RX Independently is -L RX1 -L RX2 -, where L RX1 and L RX2 Each of them is independent as described in this article.

[0184] In some implementations, -L RX1 - is -S-. In some implementations, -L RX1 - is -S-, and -S- is a Cys residue of the target-binding moiety. In some embodiments, -L RX1 - is -S-, and -S- is a Cys residue in the antibody that forms an interchain disulfide bond in the unconjugated antibody. In some embodiments, -L RX1 - is -S-, and -S- is a Cys residue of the target-binding moiety, wherein the Cys residue forms an inter-heavy chain disulfide bond in the unconjugated target-binding moiety. In some embodiments, -L RX1 - is -S-, and -S- is of the Cys residue of the target-binding moiety, wherein the Cys residue is derived by partial or complete reduction of interchain disulfide bonds.

[0185] In some implementations, L RX1 It is -NH-. In some implementations, L RX1 It is -NH-, which is a Lys residue of the target-binding moiety. In some embodiments, L RX1 It is -NH-, which is a Lys residue of the antibody or its antigen-binding fragment.

[0186] In some implementations, -L RX2- is or contains optional replacements Nitrogen and L X Bonded, and carbon with L RX1 Bonding. In some implementations, -L RX2 - Yes or contain Nitrogen and L X Bonded, and carbon with L RX1 Bonding. In some implementations, -L RX2 - Yes or contain Where "#" indicates a match with L X The connection point, and "*" indicates the connection with L. RX1 The connection point. In some implementations, -L RX2 -is-S-.

[0187] In some implementations, L RX2 It is or contains -C(O)-. In some implementations, -C(O)- is associated with L. RX1 -NH- bonds.

[0188] In some implementations, L RX Yes or contains -SS-. In some implementations, L RX It is or contains -S-CH2-C(O)-. In some implementations, L RX It is or contains optional replacements Nitrogen and L X Bonding. In some implementations, L RX yes Nitrogen and L X ("SM") bond.

[0189] In some implementations, -L RX - is or contains optional replacements Where -C(O)- and L X Bonding. In some implementations, -L RX - Yes or contain Where -C(O)- and L X Bonding.

[0190] In some implementations, L RX It is or contains -NH-C(O)-.

[0191] In some implementations, L RX It may contain a click chemical product portion. In some embodiments, L RX It may contain, or optionally contain, a substituted triazole ring. In some embodiments, L RX Yes or include In some implementations, LRX Yes or include In some implementations, L RX Yes or include In some implementations, the nitrogen in the 5-membered ring reacts with L... X Bonding. In some implementations, the carbon or nitrogen in the 8-membered ring bonds with L. X Bonding.

[0192] In some implementations, L RX It is a covalent bond. In some implementations, L RX It is composed of unattached target-binding portions (e.g., L). RXT L RX1 The groups present on (etc.) and containing L X -(CH2CH2O)nL ZP -L C - and payload portion (e.g., L) RXP L RX2 The portion produced by the reaction between groups present on the agent (etc.). In some embodiments, L RX It is composed of unattached target-binding portions (e.g., L). RXT L RX1 The groups present on (etc.) and containing L X’ and optional payload portion (e.g., L) RXP L RX2 The part produced by the reaction between groups present on the agent (etc.).

[0193] In some implementations, each L RX It is a portion that is independently produced by the reaction between unbonded joints and groups present on unbonded target-bonded parts.

[0194] In some embodiments, the adapter comprises a portion generated by a reaction between an unbound adapter and a group present on an unconjugated target-binding portion. In some embodiments, the adapter comprises a portion generated by a reaction between an unbound adapter and a group present on an antibody or its antigen-binding fragment.

[0195] In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker and a group present on arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, lysine, serine, threonine, tryptophan, or tyrosine within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker and a group present on cysteine, lysine, serine, threonine, or tyrosine within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker and a group present on cysteine ​​within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker and a group present on lysine within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker and a thiol, amine, acid, or hydroxyl group within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by the reaction between an unbound linker and a thiol within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by the reaction between an unbound linker and an amine within the target-binding portion (e.g., an antibody or its antigen-binding fragment). In some embodiments, the linker comprises a portion generated by the reaction between an unbound linker and a hydroxyl group within the target-binding portion (e.g., an antibody or its antigen-binding fragment).

[0196] In some embodiments, the linker comprises a portion resulting from a reaction between an unbound linker and a group present on a target-binding moiety (e.g., an antibody or its antigen-binding fragment), wherein the reaction is a conjugation with an activated ester, sulfonyl chloride, or isothiocyanate; reductive alkylation; aza-Michael addition; azacyclobutanone chemistry; benzoyl fluoride-based chemicals; or click chemistry. Examples are provided, for example, in Sletten and Bertozzi. Agew. Chem. Int. Ed. Engl. 2009, 48(38), 6974-6998.

[0197] In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker containing a portion capable of reacting with arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, lysine, serine, threonine, tryptophan, or tyrosine and a group present on the target binding portion (e.g., an antibody or an antigen-binding fragment thereof). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker containing a portion capable of reacting with cysteine, lysine, serine, threonine, or tyrosine and a group present on the target binding portion (e.g., an antibody or an antigen-binding fragment thereof). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker containing a portion capable of reacting with cysteine ​​and a group present on the target binding portion (e.g., an antibody or an antigen-binding fragment thereof). In some embodiments, the linker comprises a portion generated by a reaction between an unbound linker containing a portion capable of reacting with lysine and a group present on the target binding portion (e.g., an antibody or an antigen-binding fragment thereof).

[0198] In some embodiments, the portion capable of reacting with cysteine ​​is or contains maleimide, a haloacetamide (e.g., fluoroacetamide, chloroacetamide, bromoacetamide, or iodoacetamide), or a Michael acceptor (e.g., an α,β-unsaturated carbonyl group, such as acrylamide or an α,β-unsaturated ketone). In some embodiments, the portion capable of reacting with cysteine ​​is maleimide. In some embodiments, the portion capable of reacting with cysteine ​​is a haloacetamide (e.g., fluoroacetamide, chloroacetamide, bromoacetamide, or iodoacetamide). In some embodiments, the portion capable of reacting with cysteine ​​is a Michael acceptor (e.g., an α,β-unsaturated carbonyl group, such as acrylamide or an α,β-unsaturated ketone). In some embodiments, the portion capable of reacting with cysteine ​​is maleimide, chloroacetamide, bromoacetamide, acrylamide, or an α,β-unsaturated ketone. In some embodiments, the portion capable of reacting with cysteine ​​is chloroacetamide. In some embodiments, the portion capable of reacting with cysteine ​​is bromoacetamide. In some embodiments, the part that reacts with cysteine ​​is iodoacetamide. In some embodiments, the part that reacts with cysteine ​​is acrylamide. In some embodiments, the part that reacts with cysteine ​​is an α,β-unsaturated ketone.

[0199] In some implementations, the connector includes Where: # indicates the connection point with the target; and * indicates the connection point with the rest of the connector. In some embodiments, the connector includes Where R is as described herein, # indicates the connection point with the target engagement portion; and * indicates the connection point with the remainder of the connector. In some embodiments, the connector includes Where R is as described herein, # indicates the connection point with the target engagement portion; and * indicates the connection point with the remainder of the connector. In some embodiments, the connector includes , where # indicates the connection point with the target engagement portion; and * indicates the connection point with the rest of the connector. In some embodiments, the connector includes , where # indicates the connection point with the target engagement portion; and * indicates the connection point with the rest of the connector. In some embodiments, the connector includes , where * indicates the connection point with the target; and # indicates the connection point with the rest of the connector. In some embodiments, the connector includes , where # indicates the junction point with the anti-HHLA2 antibody or its antigen-binding fragment; and * indicates the junction point with the remainder of the adapter. In some embodiments, the adapter comprises Where R is as described herein, # indicates a junction point with an antibody or its antigen-binding fragment; and * indicates a junction point with the remainder of the adapter. In some embodiments, the adapter comprises Where R is as described herein, # indicates a junction point with an antibody or its antigen-binding fragment; and * indicates a junction point with the remainder of the adapter. In some embodiments, the adapter comprises , where # indicates a junction point with an antibody or its antigen-binding fragment; and * indicates a junction point with the remainder of the adapter. In some embodiments, the adapter comprises , where # indicates a junction point with an antibody or its antigen-binding fragment; and * indicates a junction point with the remainder of the adapter. In some embodiments, the adapter comprises , where * indicates the junction point with the antibody or its antigen-binding fragment; and # indicates the junction point with the rest of the adapter.

[0200] In some embodiments, the target-binding portion is engineered to incorporate reactive groups. For example, in some embodiments, an antibody or its antigen-binding fragment is engineered to incorporate Cys, Lys, etc., at certain positions for conjugation. In some embodiments, non-natural amino acid residues are incorporated, such as those containing -N3, alkynes, etc. In some embodiments, glycosylation is modified to introduce reactive groups for conjugation.

[0201] Reports include useful groups and reactions (e.g., L...). RX1 L RX2 L RXVarious techniques (etc.) can be utilized according to this disclosure, such as Fu, Z., Li, S., Han, S. et al., Antibody drug conjugate: the “biological missile” for targeted cancer therapy. Sig Transduct Target Ther7, 93 (2022). https: / / doi.org / 10.1038 / s41392-022-00947-7; Su Z, Xiao D, Xie F, Liu L, Wang Y, Fan S, Zhou X, Li S. Antibody-drug conjugates: Recent advances in linker chemistry. Acta Pharm Sin B. Dec 2021;11(12):3889-3907. doi:10.1016 / j.apsb.2021.03.042. Electronic publication on April 6, 2021. PMID: 35024314; PMCID: PMC8727783; Sheyi R, de la Torre BG, Albericio F. Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate. Pharmaceutics. 11 Feb 2022; 14(2):396. doi: 10.3390 / pharmaceutics14020396. PMID: 35214128; PMCID: PMC8874516 etc.

[0202] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L RX -L X-(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C- is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L RX -L X-(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C- It is bonded to the payload portion at its -C(O)-. In some embodiments, the -S- in -SM is a Cys residue as described herein.

[0203] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C- is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C- is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C- is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some embodiments, -L RX -L X -(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-. In some embodiments, -S- is a Cys residue as described herein.

[0204] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C- is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L RX -L X -(CH2CH2O)nLZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - is -S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-. In some embodiments, -S- is a Cys residue as described herein.

[0205] L T In some implementations, L T It is a covalent bond. In some implementations, L T It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L T The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, and -S(O)2(NR')-. In some embodiments, L T It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 Aliphatic, etc., of which L T One to five (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, and -S(O)2(NR')-. In some embodiments, L T It is a bivalent straight or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) aliphatic groups. In some embodiments, L T Is it a bivalent straight chain or branched chain C? 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 (etc.) aliphatic groups. In some embodiments, L T It is a bivalent straight chain or branched chain C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) alkylene groups. In some embodiments, L T It is a divalent linear C that can be arbitrarily replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) alkylene groups. In some embodiments, L T -(CH2) is optionally substituted. 1-5 - In some implementations, L T -(CH2) is optionally substituted.1-5 - In some implementations, L T It is -(CH2) 1-5 - In some implementations, L T It is -CH2-. In some implementations, L T It is -(CH2)4-. In some embodiments, the target-binding moiety comprises an amino acid residue, L T It bonds to the backbone atoms of amino acid residues. In some embodiments, the target-binding portion comprises amino acid residues, L T It bonds to the α-carbon of an amino acid residue. In some embodiments, L T Before conjugation, it is part of the amino acid residues (e.g., Cys, Lys, etc.) of the target binding site.

[0206] In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L T -L RX -LX -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-SM-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L T -L RX -L X-(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-. In some embodiments, the -S- in -SM is a Cys residue as described herein. In some embodiments, -CH2-S- is a Cys residue. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - It bonds to the carbon atoms (e.g., α-carbon) of the polypeptide chain and the payload portion.

[0207] In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)5C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C- is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2)3C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-GGFG-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Ala-PABA-. In some implementations, -L T -L RX -L X-(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-Val-Cit-PABA-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-oGlc-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2NHC(O)-mGlc-. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(oGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-(mGlc-a)-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-mGlu-. In some implementations, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - is -CH2-S-CH2C(O)NH-(CH2CH2O)8-(CH2)2C(O)-oGlu-. In some implementations, -L T -L RX -LX -(CH2CH2O)nL ZP -L C - It is bonded to the payload portion at its -C(O)-. In some embodiments, -S- is a Cys residue as described herein. In some embodiments, -CH2-S- is a Cys residue. In some embodiments, -L T -L RX -L X -(CH2CH2O)nL ZP -L C - It bonds to the carbon atoms (e.g., α-carbon) of the polypeptide chain and the payload portion.

[0208] L X' In some implementations, the connector is or includes L X In some implementations, L X 'It is a covalent bond.' In some implementations, L X ' is a divalent or multivalent straight or branched chain C that can be optionally replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C 2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L X The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-; -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X ' is a divalent or multivalent straight or branched chain C that can be optionally replaced. 1-20 (For example, C) 1-6 C 1-10 C 1-15 C2-8 C 5-10 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 C 20 Aliphatic, etc., of which L X The 1-10 methylene units (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, 1-3, 1-2, etc.) are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-; -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X ' is a divalent or multivalent straight or branched chain C that can be optionally replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 Aliphatic, etc., of which L X’ One to five (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-; -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X It is bivalent. In some implementations, L X It is multi-valent. For example, in some implementations, L X’ It was arbitrarily replaced. The benzene ring is bonded to -(CH2CH2O)n-. In some embodiments, have The structure. In some implementations, Having structure In some embodiments, each -S- is independently a Cys residue in the same target-binding moiety. In some embodiments, each -CH2-S- is independently a Cys residue in the same target-binding moiety. In some embodiments, two Cys residues form an interchain disulfide bond in the unconjugated target-binding moiety.

[0209] In some implementations, a is 1. In some implementations, a is 2-10. In some implementations, a is 2. In some implementations, a is 3, 4, 5, 6, 7, 8, 9, or 10.

[0210] In some implementations, b is 1. In some implementations, b is 2-10. In some implementations, b is 2. In some implementations, b is 3, 4, 5, 6, 7, 8, 9, or 10.

[0211] Cy As used herein, Cy is a divalent 3-20-membered ring having 0-10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-10, 0-5, 1-5, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, with optional substitutions. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, Cy is a divalent 3-20-membered ring having 0-10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-10, 0-5, 1-5, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur, with optional substitutions. In some embodiments, Cy is monocyclic. In some embodiments, Cy is bicyclic. In some embodiments, Cy is polycyclic. In some embodiments, each monocyclic unit is independently a 3-10 membered (e.g., 3, 4, 5, 1-5, 1-4, 1-2, etc.) ring having 0-5 (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, and 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, etc.) carbon atoms optionally substituted. In some embodiments, the monocyclic unit is an optionally substituted saturated ring. In some embodiments, the monocyclic unit is an optionally substituted partially unsaturated ring. In some embodiments, the monocyclic unit is an optionally substituted aromatic ring. In some embodiments, the monocyclic unit is an optionally substituted benzene ring. In some embodiments, the monocyclic unit is a 5-membered heteroaromatic compound having 1-4 (e.g., 1, 2, 3, 4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, optionally substituted. In some embodiments, the monocyclic unit is a 6-membered heteroaromatic compound having 1-4 (e.g., 1, 2, 3, 4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, optionally substituted. Each monocyclic unit is independently and optionally substituted. In some embodiments, the monocyclic unit is unsaturated. In some embodiments, the monocyclic unit is saturated. In some embodiments, Cy is an optionally substituted saturated ring. In some embodiments, Cy is an optionally substituted partially unsaturated ring. In some embodiments, Cy is an optionally substituted aromatic ring.

[0212] In some embodiments, Cy is an optionally substituted divalent 3-10 quinone (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 quinone) monocyclic alicyclic ring. In some embodiments, Cy is an optionally substituted 3-10 quinone (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 quinone) monocyclic cycloalkyl ring. In some embodiments, Cy is an optionally substituted 3-10 quinone (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 quinone) monocyclic heteroalicyclic ring having 1-5 heteroatoms. In some embodiments, Cy is an optionally substituted 3-10 quinone (e.g., 3, 4, 5, 6, 7, 8, 9, or 10 quinone) monocyclic heteroalkyl ring having 1-5 heteroatoms. In some embodiments, Cy is an optionally substituted divalent 5-15 quintone (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 quintone) bicyclic or polycyclic alicyclic group. In some embodiments, Cy is an optionally substituted divalent 5-15 quintone (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 quintone) bicyclic or polycyclic cycloalkyl group. In some embodiments, Cy is an optionally substituted 5-15 quintone (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 quintone) bicyclic or polycyclic heteroalicyclic ring having 1-5 heteroatoms. In some embodiments, Cy is a optionally substituted 5-15 membered (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 membered) bicyclic or polycyclic heterocyclic ring having 1-5 heteroatoms. In some embodiments, the alicyclic, cycloalkyl, heteroalicyclic, or heteroalkyl ring is 3 membered. In some embodiments, it is 4 membered. In some embodiments, it is 5 membered. In some embodiments, it is 6 membered. In some embodiments, it is 7 membered. In some embodiments, it is 8 membered. In some embodiments, it is 9 membered. In some embodiments, it is 10 membered. In some embodiments, it is 11 membered. In some embodiments, it is 12 membered. In some embodiments, Cy is an optionally substituted phenylene. In some embodiments, Cy is an optionally substituted divalent 10 membered bicyclic aryl ring. In some embodiments, Cy is a 5-membered heteroaryl ring with 1-4 heteroatoms that is optionally substituted. In some embodiments, Cy is a 6-membered heteroaryl ring with 1-4 heteroatoms that is optionally substituted. In some embodiments, Cy is a 9-membered bicyclic heteroaryl ring with 1-5 heteroatoms that is optionally substituted. In some embodiments, Cy is a 10-membered bicyclic heteroaryl ring with 1-5 heteroatoms that is optionally substituted. In some embodiments, the heteroaliphatic, heterocyclic, or heteroaryl ring contains no more than one heteroatom. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur.

[0213] In some embodiments, Cy is an optionally substituted 4-7 membered ring having 0-3 heteroatoms. In some embodiments, Cy is an optionally substituted 6-membered aryl ring. In some embodiments, the aryl ring is substituted. In some embodiments, it is substituted with one or more halogens. In some embodiments, it is substituted with one or more -F atoms. In some embodiments, it is unsubstituted. In some embodiments, it is optionally substituted. In some implementations, it is In some implementations, it is optionally replaced. In some implementations, it is In some implementations, it is optionally replaced. In some implementations, it is In some embodiments, Cy is an optionally substituted 5-membered heteroaryl ring having 1-3 heteroatoms. In some embodiments, the heteroatom is nitrogen. In some embodiments, the heteroatom is oxygen. In some embodiments, the heteroatom is sulfur. In some embodiments, Cy is optionally substituted. In some implementations, Cy is In some implementations, -Cy- is optionally replaced. In some implementations, -Cy- is In some implementations, -Cy- is In some implementations, -Cy- is optionally replaced. In some implementations, -Cy- is optionally replaced. In some implementations, -Cy- is .

[0214] R’ The following describes some implementations of R'. Various variables can be R', and implementations of R' can also be applied to such variables that can be R'.

[0215] In some embodiments, R' is the R described herein. In some embodiments, R' is -H. In some embodiments, R' is not -H. In some embodiments, R' is C, which is optionally replaced. 1-6 Lipids.

[0216] In some embodiments, R' is -C(O)R, where R is as described herein. In some embodiments, R' is -CO2R, where R is as described herein. In some embodiments, R' is -S(O)2R, where R is as described herein. In some embodiments, as described herein, R is not -H.

[0217] In some embodiments, two or more R's together with their intermediate atoms form optionally substituted 3-20 denominator rings (e.g., 3-15, 3-10, 3-6, 5-10, 5-6, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 denominator rings, which, in addition to the intermediate atom, also have 0-10 heteroatoms (e.g., 0, 1-10, 1-5, 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, two or more R's on the same atom, together with the atoms to which they are attached, form optionally substituted 3-20 denominator rings (e.g., 3-15, 3-10, 3-6, 5-10, 5-6, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 denominator rings), which, in addition to the intermediate atom, have 0-10 heteroatoms (e.g., 0, 1-10, 1-5, 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, two R's on the same atom, together with the atoms they are attached to, form optionally substituted 3-20 denominator rings (e.g., 3-15, 3-10, 3-6, 5-10, 5-6, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 denominator rings, which, in addition to the intermediate atom, have 0-10 heteroatoms (e.g., 0, 1-10, 1-5, 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur.

[0218] In some embodiments, each R' is independently -R, -C(O)R, -CO2R or -S(O)2R, or two or more R's on the same atom together with the atoms to which they are attached form an optionally substituted 3-20 membered ring, which, in addition to the intermediate atom, has 0-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0219] ring The compounds disclosed herein (e.g., in compositions and / or conjugates as described herein) may contain various rings. In some embodiments, the variable (e.g., R or a variable that may be R) may have a ring as described herein. In some embodiments, two variables (e.g., two R' groups) may form a ring as described herein together with their intermediate atoms. In some embodiments, the ring is monovalent. In some embodiments, the ring is divalent. In some embodiments, the ring structure may be polyvalent. The ring may optionally be substituted. In some embodiments, the ring is unsubstituted. In some embodiments, the ring is substituted.

[0220] The following describes some implementation schemes and features of the ring as examples.

[0221] In some embodiments, the ring is a 3-20 member (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1-10, 0-5, 1-5, etc.) ring having 0-10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3-15, 3-14, 3-10, 5-10, 3-7, 3-6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the ring is a 3-20 member (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 1-10, 0-5, 1-5, etc.) ring having 0-10 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 3-15, 3-14, 3-10, 5-10, 3-7, 3-6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur, with optional substitutions. In some embodiments, the ring is monocyclic. In some embodiments, the ring is bicyclic. In some embodiments, the ring is polycyclic.

[0222] In some embodiments, each monocyclic unit is independently a 3-10 membered (e.g., 3, 4, 5, 1-5, 1-4, 1-2, etc.) ring having 0-5 (e.g., 1, 2, 3, 4, 5, 1-5, 1-4, 1-2, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, and 1-10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1-5, 1-4, etc.) carbon atoms optionally substituted. In some embodiments, the monocyclic unit is an optionally substituted saturated ring. In some embodiments, the monocyclic unit is an optionally substituted partially unsaturated ring. In some embodiments, the monocyclic unit is an optionally substituted aromatic ring. In some embodiments, the monocyclic unit is an optionally substituted benzene ring. In some embodiments, the monocyclic unit is a 5-membered heteroaromatic compound having 1-4 (e.g., 1, 2, 3, 4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, optionally substituted. In some embodiments, the monocyclic unit is a 6-membered heteroaromatic compound having 1-4 (e.g., 1, 2, 3, 4, 1-3, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, optionally substituted. Each monocyclic unit is independently and optionally substituted. In some embodiments, the monocyclic unit is unsaturated. In some embodiments, the monocyclic unit is saturated.

[0223] In some embodiments, the ring is a saturated ring that is optionally substituted. In some embodiments, the ring is a partially unsaturated ring that is optionally substituted. In some embodiments, the ring is an aromatic ring that is optionally substituted.

[0224] In some embodiments, the ring is an optionally substituted 3-10 member monocyclic, bicyclic, or polycyclic ring, having 0-4 heteroatoms independently selected from nitrogen, oxygen, and sulfur in addition to one or more intermediate atoms. In some embodiments, the ring is 3-9, 3-8, 3-7, 3-6, 4-10, 4-9, 4-8, 4-7, 4-6, 5-10, 5-9, 5-8, 5-7, 5-6 membered or 3, 4, 5, 6, 7, 8, 9, or 10 membered. In some embodiments, the ring is 3-9 membered. In some embodiments, the ring is 3-7 membered. In some embodiments, the ring is 4-10 membered. In some embodiments, the ring is 4-7 membered. In some embodiments, the ring is 5-10 membered. In some embodiments, the ring is 5-7 membered. In some embodiments, the ring is 3 membered. In some embodiments, the ring is 4 membered. In some embodiments, the ring is 5 membered. In some embodiments, the ring is hexacyclic. In some embodiments, the ring is septium-cyclic. In some embodiments, the ring is octium-cyclic. In some embodiments, the ring is quintium-cyclic. In some embodiments, the ring is decacyclic. In some embodiments, the ring is monocyclic. In some embodiments, the ring is bicyclic. In some embodiments, the ring is polycyclic. In some embodiments, the ring does not have heteroatoms. In some embodiments, the ring has 1-6, for example 1-5, 1-3, or 1, 2, 3, 4, 5, or 6 heteroatoms. In some embodiments, the ring formed by two or more groups with their intermediate atoms has no heteroatoms other than the intermediate atoms. In some embodiments, the ring formed by two or more groups with one or more of their intermediate atoms has 1-6, for example 1-5, 1-3, or 1, 2, 3, 4, 5, or 6 heteroatoms other than one or more intermediate atoms. In some embodiments, the ring is saturated. In some embodiments, the ring is partially unsaturated. In some embodiments, the ring comprises one or more aromatic rings. In some embodiments, the ring is bicyclic or polycyclic, and each monocyclic unit is independently 3-10 quinary (e.g., 3-9, 3-8, 3-7, 3-6, 4-10, 4-9, 4-8, 4-7, 4-6, 5-10, 5-9, 5-8, 5-7, 5-6 or 3, 4, 5, 6, 7, 8, 9 or 10 quinary), saturated, partially unsaturated, or aromatic, and has 0-5 (e.g., 0, 1-5, 1-3, 1, 2, 3, 4 or 5) heteroatoms. In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, the monocyclic unit is saturated. In some embodiments, the monocyclic unit is partially unsaturated. In some embodiments, the monocyclic unit is aromatic. In some embodiments, the monocyclic unit is heteroaromatic. Those skilled in the art will understand that if multiple paths exist, the intermediate atoms of the groups that form the ring together are typically atoms on the shortest path connecting these groups.

[0225] In some embodiments, the ring has one or more heteroatoms. In some embodiments, the ring contains a nitrogen atom. In some embodiments, the ring contains an oxygen atom. In some embodiments, the ring contains a sulfur atom.

[0226] R The various variables in this disclosure can be R independently, and embodiments of R can also be applied to such variables that can be R. Some embodiments of R are described below as examples. Those skilled in the art will understand that embodiments described for variables that can be R and fall within the definition of R can also be embodiments of R. Those skilled in the art will further understand that embodiments of R (e.g., those described for R, those described for variables that can be R and fall within the definition of R, etc.) can be embodiments of variables that can be R.

[0227] In some implementations, R is -H. In some implementations, R is not -H. In some implementations, R is C, which is optionally replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) aliphatic. In some embodiments, R is optionally replaced by C. 1-8 Aliphatic. In some embodiments, R is C, which is optionally replaced. 1-6 Aliphatic. In some embodiments, R is C, which is optionally replaced. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) alkyl. In some embodiments, R is optionally substituted C. 1-8 Alkyl group. In some embodiments, R is an optional substituted C. 1-6 Aliphatic. In some embodiments, R is C, which is optionally replaced. 1-6 Alkyl group. In some embodiments, R is optionally substituted with -CH2-C. 3-6 Alicyclic compound. In some embodiments, R is optionally substituted with -CH2-C. 3-6 Alicyclic compound. In some embodiments, R is optionally substituted with -CH2-C. 3-6Cycloalkyl. In some embodiments, R is methyl. In some embodiments, R is ethyl. In some embodiments, R is isopropyl. In some embodiments, R is -CF3. In some embodiments, R is -CH2CF3. In some embodiments, R is butyl. In some embodiments, R is tert-butyl.

[0228] In some embodiments, R is an optional 3-10 quinone (e.g., 3-9, 3-8, 3-7, 3-6, 5-7, 4, 5, 6, 7, 8, 9, 10 quinones, etc.) alicyclic compound. In some embodiments, R is an optional substituted C. 3-10 Cycloalkyl. In some embodiments, R is optionally substituted cyclopropyl. In some embodiments, R is optionally substituted cyclobutyl. In some embodiments, R is optionally substituted cyclopentyl. In some embodiments, R is optionally substituted cyclohexyl.

[0229] In some embodiments, R is a C that is optionally substituted with 1 to 5 heteroatoms (e.g., 1 to 4, 1 to 3, 1 to 2, 1, 2, 3, 4, 5, etc.) independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) heteroaliphatic. In some embodiments, R is a C that is optionally substituted with 1-5 (e.g., 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-6 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) heteroaliphatic. In some embodiments, R is a C that is optionally substituted with 1-5 (e.g., 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. 1-10 (For example, 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C 10 (etc.) heteroaliphatic. In some embodiments, R is a C that is optionally substituted with 1-5 (e.g., 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur. 1-6 (For example, C) 1-6 C 2-8 ,C1,C2,C3,C4,C5,C6,C7,C8,C9,C10 (etc.) heteroaliphatic. In some embodiments, R is a C1-6 heteroaliphatic group having 1-3 independently selected heteroatoms, optionally substituted; or a C1-6 heteroaliphatic group having 1-3 independently selected heteroatoms, independently selected from nitrogen, oxygen, and sulfur. In some embodiments, R is a C1-6 heteroaliphatic group having 1-3 independently selected heteroatoms, optionally substituted; or a C1-6 heteroaliphatic group having 1 independently selected heteroatom. In some embodiments, R is a heteroalkyl group.

[0230] In some embodiments, R is a 3-15 membered (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 3-10, 3-7, 4-10, 4-6 membered, etc.) heterocycle having 1-5 (e.g., 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, with optional substitution. In some embodiments, R is a 3-15 membered (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 3-10, 3-7, 4-10, 4-6 membered, etc.) heterocycle having 1-5 (e.g., 1-4, 1-3, 1-2, 1, 2, 3, 4, 5, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur, with optional substitution. In some embodiments, R is a 3- to 10-membered heterocyclic group (e.g., 3-9, 3-8, 3-7, 3-6, 5-7, 4, 5, 6, 7, 8, 9, 10, etc.) having 1-4 (e.g., 1, 2, 3, 4, 1-2, etc.) heteroatoms independently selected from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 3-membered heterocyclic group having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 3-membered heterocyclic group having one heteroatom independently selected from nitrogen, oxygen, and sulfur, which is optionally substituted. In some embodiments, R is a 4-membered heterocyclic group having 1-2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 5-membered heterocyclic group having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 6-membered heterocyclic group having 1-3 independently selected heteroatoms, optionally substituted, selected from nitrogen, oxygen, and sulfur. In some embodiments, R is a 7-membered heterocyclic group having 1-3 independently selected heteroatoms, optionally substituted, selected from nitrogen, oxygen, and sulfur. In some embodiments, R is an 8-membered heterocyclic group having 1-3 independently selected heteroatoms, optionally substituted, selected from nitrogen, oxygen, and sulfur. In some embodiments, R is a 9-membered heterocyclic group having 1-3 independently selected heteroatoms, optionally substituted, selected from nitrogen, oxygen, and sulfur. In some embodiments, R is a 10-membered heterocyclic group having 1-3 independently selected heteroatoms, optionally substituted, selected from nitrogen, oxygen, and sulfur. In some embodiments, a carbon atom is present in the heterocyclic ring. In some embodiments, two or more carbon atoms (e.g., 2-14, 2-9, 2-6, 2-4, 3-10, 3-5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, etc.) are present in the heterocyclic base ring.

[0231] In some embodiments, R is an optionally substituted phenyl group. In some embodiments, R is a phenyl group. In some embodiments, R is an optionally substituted naphthyl group. In some embodiments, R is an optionally substituted 1-naphthyl group. In some embodiments, R is an optionally substituted 2-naphthyl group. In some embodiments, R is a naphthyl group.

[0232] In some embodiments, R is a 5-14 membered (e.g., 5, 6, 9, 10, 14, etc.) heteroaryl group having 1-6 (e.g., 1-5, 1-4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur, which are optionally substituted. In some embodiments, R is a 5-14 membered (e.g., 5, 6, 9, 10, 14, etc.) heteroaryl group having 1-6 (e.g., 1-5, 1-4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur, which are optionally substituted. In some embodiments, R is a 5-10 membered (e.g., 5, 6, 9, 10, etc.) heteroaryl group having 1-6 (e.g., 1-5, 1-4, 1, 2, 3, 4, 5, 6, etc.) heteroatoms independently selected from oxygen, nitrogen, and sulfur, which are optionally substituted. In some embodiments, R is a 5-10-membered heteroaryl group having 1-4 independently selected heteroatoms chosen from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, the heteroaryl ring is monocyclic and 5-membered. In some embodiments, the heteroaryl ring is monocyclic and 6-membered. In some embodiments, the heteroaryl ring is bicyclic and 9-membered. In some embodiments, the heteroaryl ring is bicyclic and 10-membered. In some embodiments, the heteroaryl ring is tricyclic and 14-membered. In some embodiments, R is a 5-membered heteroaryl group having 1-4 independently selected heteroatoms chosen from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 5-membered heteroaryl group having 1 independently selected heteroatomium, oxygen, and sulfur, which is optionally substituted. In some embodiments, R is a 6-membered heteroaryl group having 1-4 independently selected heteroatoms chosen from nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a 6-membered heteroaryl group having one heteroatom selected from nitrogen, oxygen, and sulfur, which is optionally substituted. In some embodiments, R is a bicyclic 8-10-membered aromatic ring having 1-6 heteroatoms of nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a bicyclic 9-membered aromatic ring having 1-6 heteroatoms of nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a bicyclic 10-membered aromatic ring having 1-6 heteroatoms of nitrogen, oxygen, and sulfur, which are optionally substituted. In some embodiments, R is a bicyclic 9-membered aromatic ring having one heteroatom selected from nitrogen, oxygen, and sulfur, which is optionally substituted. In some embodiments, R is a bicyclic 10-membered aromatic ring having one heteroatom of nitrogen, oxygen, and sulfur, which is optionally substituted. In some embodiments, at least one heteroatom is nitrogen. In some embodiments, at least one heteroatom is oxygen. In some embodiments, at least one heteroatom is sulfur. In some embodiments, each heteroatom is the same. In some embodiments, at least one heteroatom is different from the other heteroatom.

[0233] In some implementations, multiple divalent portions exist between the two connected portions. As those skilled in the art will understand, when each of these divalent portions is a covalent bond, a single covalent bond exists between the connected portions unless otherwise stated.

[0234] As described herein, various groups can be optionally substituted. Substituents are conventionally used in chemistry, including for the development of various therapeutic agents. Many substituents can be utilized according to this disclosure. In some embodiments, the optionally substituted group is unsubstituted. In some embodiments, the optionally substituted group is substituted. Substituents are preferably those that result in the formation of compounds having the desired properties, activities, uses, etc., as described herein. In some embodiments, the compound is stable for therapeutic uses as described herein. As used herein, the term "stable" means a compound that is substantially unchanged when subjected to conditions permissible for its production, testing, and, in some embodiments, its recovery, purification, and use for one or more purposes disclosed herein. In some embodiments, the substituent is a hydrocarbon group. In some embodiments, the substituent contains heteroatoms. In some embodiments, the substituent contains multiple heteroatoms. In some embodiments, each atom in the substituent is independently selected from hydrogen, carbon, halogen, nitrogen, oxygen, sulfur, phosphorus, and silicon. In some embodiments, each atom in the substituent is independently selected from hydrogen, carbon, halogen, nitrogen, oxygen, and sulfur. In some embodiments, each atom in the substituent is independently selected from hydrogen, carbon, fluorine, chlorine, bromine, iodine, nitrogen, oxygen, and sulfur. In some embodiments, the total number of carbon atoms and non-halogenated heteroatoms in the substituent is about or no more than about 1; in some embodiments, it is no more than about 2; in some embodiments, it is no more than about 3; in some embodiments, it is no more than about 4; in some embodiments, it is no more than about 5; in some embodiments, it is no more than about 6; in some embodiments, it is no more than about 7; in some embodiments, it is no more than about 8; in some embodiments, it is no more than about 9; in some embodiments, it is no more than about 10; in some embodiments, it is no more than about 11; in some embodiments, it is no more than about 12; in some embodiments, it is no more than about 13; in some embodiments, it is no more than about 14; in some embodiments, it is no more than about 15; in some embodiments, it is no more than about 20. In some embodiments, the total number of carbon atoms and non-halogenated heteroatoms in each substituent is independently no more than about 20. In some embodiments, the total number of carbon atoms and non-halogenated heteroatoms in each substituent is independently no more than about 15. In some embodiments, the total number of carbon atoms and nonhalogenated heteroatoms in each substituent independently does not exceed about 10. In some embodiments, the total number of carbon atoms and nonhalogenated heteroatoms in each substituent independently does not exceed about 6.

[0235] Decomposable unit In some embodiments, the connector includes a pyrolytic unit. In some embodiments, L C1 It is a pyrolytic unit.

[0236] In some embodiments, the cleavable unit is a chemically cleavable unit. Such cleavable units are cleavable under certain chemical conditions without enzymatic catalysis. In some embodiments, the cleavable unit is an acid-cleavable unit. In some embodiments, the cleavable unit is or contains an hydrazone group. In some embodiments, the cleavable unit is or contains a carbonate group. In some embodiments, the cleavable unit is or contains a silyl group. In some embodiments, the cleavable unit is or contains -SS-. In some embodiments, the cleavable unit is cleavable in the presence of GSH. In some embodiments, the cleavable unit is cleavable in the presence of Fe(II). In some embodiments, the cleavable unit is or contains a 1,2,4-trioxacyclopentane moiety.

[0237] In some embodiments, the cleavable unit is an enzyme-cleavable unit (i.e., a unit or portion thereof that is cleaved by an enzyme). In some embodiments, the enzyme-cleavable unit comprises an amino acid sequence that is cleaved by a protease (e.g., an intracellular protease). In some embodiments, the protease breaks the amide bonds between the amino acid sequences within the cleavable unit. In some embodiments, the enzyme-cleavable unit comprises an amino acid sequence that is cleaved by a protease (e.g., an intracellular human protease). In some embodiments, the enzyme-cleavable unit comprises an amino acid sequence that is cleaved by a cathepsin (e.g., human cathepsin L, B, F, or S).

[0238] In some embodiments, the enzyme-cleavable unit contains between 1 and 12 amino acids. In some embodiments, the enzyme-cleavable unit contains between 2 and 6 amino acids. In some embodiments, the enzyme-cleavable unit contains between 2 and 5 amino acids. In some embodiments, the enzyme-cleavable unit contains between 2 and 4 amino acids. In some embodiments, the enzyme-cleavable unit contains 2 or 3 amino acids. In some embodiments, the enzyme-cleavable unit contains 3 or 4 amino acids. In some embodiments, the enzyme-cleavable unit contains 2 amino acids. In some embodiments, the enzyme-cleavable unit contains 3 amino acids. In some embodiments, the enzyme-cleavable unit contains 4 amino acids. In some embodiments, the enzyme-cleavable unit contains 5 amino acids. In some embodiments, the enzyme-cleavable unit contains 6 amino acids.

[0239] In some embodiments, the enzymatically cleavable unit is -DGGFG-, -KGGFG-, -GGFG-, -GGFGG-, -GGFGGG-, -KGGF-, -DGGF-, -V-Cit-, -VA-, -FK-, -AA-, -VQ-, -LQ-, -FQ-, -TT-, -AAA-, -EA-, -YR-, -FR-, -YM-, -VR-, -MT-, -VT-, -VM-, -LM-, -AN-, -GGR-, -D-Val-D-Gln-, -D-Ala-D-Ala-, -DVA-, or -FM-. In some embodiments, the enzymatically cleavable unit is selected from the following amino acid sequences: -V-Cit-, -VA-, or -AA-. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -GGFG-, -V-Cit-, -AAA-, or -DVA-. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -DGGFG-, -KGGFG-, -GGFG-, -GGFGG-, -GGFGGG-, -KGGF-, or -DGGF-. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -V-Cit-, -VA-, VQ-, -VR-, -VT-, -VM-, or -D-Val-D-Gln-. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -GGFG- or -VA-. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -VQ- or -LQ. In some embodiments, the enzyme-cleavable unit is selected from the following amino acid sequences: -LQ-, -FQ-, -TT-, -AAA-, or -EA-.

[0240] In some embodiments, the enzyme-cleavable unit is -DGGFG-. In some embodiments, the enzyme-cleavable unit is -KGGFG-. In some embodiments, the enzyme-cleavable unit is -GGFG-. In some embodiments, the enzyme-cleavable unit is -GGFGG-. In some embodiments, the enzyme-cleavable unit is -GGFGGG-. In some embodiments, the enzyme-cleavable unit is -KGGF-. In some embodiments, the enzyme-cleavable unit is -DGGF-. In some embodiments, the enzyme-cleavable unit is -V-Cit-. In some embodiments, the enzyme-cleavable unit is -VA-. In some embodiments, the enzyme-cleavable unit is -FK-. In some embodiments, the enzyme-cleavable unit is -AA-. In some embodiments, the enzyme-cleavable unit is -VQ-. In some embodiments, the enzyme-cleavable unit is -YR-. In some embodiments, the enzyme-cleavable unit is -FR-. In some embodiments, the enzyme-cleavable unit is -YM-. In some embodiments, the enzyme-cleavable unit is -VR-. In some embodiments, the enzyme-cleavable unit is -MT-. In some embodiments, the enzyme-cleavable unit is -VT-. In some embodiments, the enzyme-cleavable unit is -VM-. In some embodiments, the enzyme-cleavable unit is -LM-. In some embodiments, the enzyme-cleavable unit is -AN-. In some embodiments, the enzyme-cleavable unit is -GGR-. In some embodiments, the enzyme-cleavable unit is -D-Val-D-Gln-. In some embodiments, the enzyme-cleavable unit is -D-Ala-D-Ala-. In some embodiments, the enzyme-cleavable unit is -FM-. In some embodiments, the enzyme-cleavable unit is -GGFG- or -VA-.

[0241] In some embodiments, the cleavable unit is cleavable in the presence of a glycosidase. In some embodiments, the cleavable unit is cleavable in the presence of β-glucuronidase (e.g., in lysosomes). In some embodiments, the cleavable unit is or comprises a β-glucuronide moiety. In some embodiments, the cleavable unit is cleavable in the presence of β-galactosidase (e.g., in lysosomes). In some embodiments, the cleavable unit is or comprises a galactoside moiety.

[0242] In some embodiments, the cleavable unit is a phosphatase-cleavable unit. In some embodiments, the cleavable unit is a pyrophosphate-cleavable unit. In some embodiments, the cleavable unit is or contains pyrophosphate. In some embodiments, the cleavable unit is cleavable in the presence of phosphatase or pyrophosphate (e.g., in lysosomes).

[0243] In some embodiments, the cleavable unit is a sulfatase-cleavable unit. In some embodiments, the cleavable unit is or comprises an aryl sulfate moiety. In some embodiments, the cleavable unit is cleavable in the presence of a sulfatase (e.g., in a lysosome).

[0244] In some embodiments, the cleavable unit is a photoresponsive cleavable unit. In some embodiments, the cleavable unit is or comprises a heptamethrin fluorophore moiety. In some embodiments, it can be cleaved by irradiation with NIR light (λ = 650 nm–900 nm). In some embodiments, the cleavable unit is or comprises an o-nitrobenzyl moiety. In some embodiments, it can be cleaved by irradiation with UV light (λ = 365 nm). In some embodiments, the cleavable unit is or comprises a PC4AP moiety. In some embodiments, it can be cleaved by irradiation with near-infrared (NIR) light (λ = 365 nm) and by intramolecular addition reaction with a nearby amine.

[0245] In some embodiments, the cleavable unit is a bioorthogonal cleavable unit. In some embodiments, it is or includes a dsProc portion. In some embodiments, it can be bioorthogonally cleaved against Cu(I)-BTTAA / dsProc.

[0246] Various other cleavable units are reported and can be utilized according to this disclosure, such as Fu, Z., Li, S., Han, S. et al. Antibody drug conjugate: the “biological missile” for targeted cancer therapy. Sig Transduct Target Ther 7, 93 (2022). https: / / doi.org / 10.1038 / s41392-022-00947-7; Su Z, Xiao D, Xie F, Liu L, Wang Y, Fan S, Zhou X, Li S. Antibody-drug conjugates: Recent advances in linker chemistry. ActaPharm Sin B. Dec 2021;11(12):3889-3907. doi: 10.1016 / j.apsb.2021.03.042.epublished on April 6, 2021. PMID: 35024314; PMCID: PMC8727783; Sheyi R, de la TorreBG, Albericio F. Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate. Pharmaceutics. Feb 11, 2022; 14(2):396. doi: 10.3390 / pharmaceutics14020396. PMID: 35214128; PMCID: PMC8874516 etc.

[0247] Spacer part In some embodiments, the connector includes a spacer portion (e.g., at least one spacer portion). In some embodiments, the spacer portion comprises optionally substituted divalent C1-12 aliphatic, aryl, heteroaryl, polyether, polyester, poly(N-(2-hydroxypropyl)methacrylamide) (pHPMA), polylactic-co-glycolic acid (PLGA), alkylene diamine polymers, polysarcosine, sugars, or combinations thereof. In some embodiments, the spacer portion comprises optionally substituted divalent polyether, polysarcosine, sugars, or combinations thereof.

[0248] In some embodiments, the spacer portion contains optionally substituted divalent C. 1-10 Aliphatic. In some embodiments, the spacer portion comprises optionally substituted divalent C. 1-8 Aliphatic. In some embodiments, the spacer portion is optionally replaced with divalent C. 1-6 Aliphatic. In some embodiments, the spacer portion comprises optionally substituted divalent C. 2-6 Aliphatic. In some embodiments, the spacer portion comprises optionally substituted divalent C. 3-6 Aliphatic. In some embodiments, the spacer portion comprises optionally substituted divalent C. 4-6 Aliphatic. In some embodiments, the spacer portion comprises a divalent C5 or C6 aliphatic compound that has been optionally substituted. In some embodiments, the spacer portion comprises a divalent C3 aliphatic compound that has been optionally substituted.

[0249] In some embodiments, the spacer portion comprises a structure -(CH2CH2O). n- of polyethylene glycol, where n is an integer between 0 and 24. In some embodiments, n is an integer between 1 and 24. In some embodiments, n is an integer between 1 and 12. In some embodiments, n is an integer between 1 and 11. In some embodiments, n is an integer between 1 and 10. In some embodiments, n is an integer between 1 and 9. In some embodiments, n is an integer between 1 and 8. In some embodiments, n is an integer between 2 and 8. In some embodiments, n is an integer between 3 and 8. In some embodiments, n is an integer between 4 and 8. In some embodiments, n is an integer between 6 and 8. In some embodiments, n is 7 or 8. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10. In some implementations, n is 11. In some implementations, n is 12.

[0250] In some embodiments, the spacer portion comprises polysarcosine having the following structure: Where n is as described in this article.

[0251] In some embodiments, the spacer portion comprises a monosaccharide, disaccharide, or oligosaccharide. In some embodiments, the spacer portion comprises a monosaccharide. In some embodiments, the spacer portion comprises a glycoside. In some embodiments, the spacer portion comprises a uronic acid or a derivative thereof. In some embodiments, the spacer portion comprises glucuronic acid or a derivative thereof. In some embodiments, the spacer portion comprises glucuronic acid.

[0252] In some implementations, the spacer portion is optionally replaced. In some implementations, the spacer sub-part is .

[0253] In some implementations, the connector is or includes a structure of type A, B, or C: Or the structure of a pharmaceutically acceptable salt thereof, wherein: # indicates a junction with the target binding region (e.g., an anti-HHLA2 antibody or an antigen-binding fragment); % indicates the point of connection with the payload portion (e.g., ethacratecan or its derivatives or analogs, or a pharmaceutically acceptable salt thereof); X is a portion generated by the reaction of an unbound linker with a group present on an anti-HHLA2 antibody or its antigen-binding fragment (e.g., as described herein); Y is a cleavable unit as described herein (e.g., an enzyme-cleavable unit); and Z is the spacer sub-part as described in this paper.

[0254] Various other spacer components are reported and can be utilized according to this disclosure, such as Fu, Z., Li, S., Han, S. et al. Antibody drug conjugate: the “biological missile” for targeted cancer therapy. Sig Transduct Target Ther 7, 93 (2022). https: / / doi.org / 10.1038 / s41392-022-00947-7; Su Z, Xiao D, Xie F, Liu L, Wang Y, Fan S, Zhou X, Li S. Antibody-drug conjugates: Recent advances in linker chemistry. ActaPharm Sin B. Dec 2021;11(12):3889-3907. doi: 10.1016 / j.apsb.2021.03.042.epublished on April 6, 2021. PMID: 35024314; PMCID: PMC8727783; Sheyi R, de la TorreBG, Albericio F. Linkers: An Assurance for Controlled Delivery of Antibody-Drug Conjugate. Pharmaceutics. Feb 11, 2022; 14(2):396. doi: 10.3390 / pharmaceutics14020396. PMID: 35214128; PMCID: PMC8874516 etc.

[0255] In some implementations, the connector is or includes -L RX -L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described herein. In some implementations, the connector is or contains -L T -L RX-L X -(CH2CH2O)nL ZP -L C - where each variable is independent as described herein. In some implementations, the connector is or includes -SM1-L X -(CH2CH2O)nL ZP -L C - where SM1 is The carbon atom is bonded to the target-binding portion (e.g., an antibody), and each variable is independent as described herein. In some embodiments, the linker is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-L C - where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-L C - where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-GGFG-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-GGFG-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-Val-Ala-PABA-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-Val-Ala-PABA-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z-C(O)-Val-Cit-PABA-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-Val-Cit-PABA-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -N(R')-C(O)-oGlu-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -NH-C(O)-oGlu-, where each variable is independent as described herein. In some embodiments, the connector is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -N(R')-C(O)-mGlu-, where each variable is independent as described herein. In some implementations, the connector is or contains -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -NH-C(O)-mGlu-, where each variable is independent as described herein. In some embodiments, x is 2-10. In some embodiments, n is 2-10. In some embodiments, z is 2-10. In some embodiments, x is 2, n is 8, and z is 2. In some embodiments, x is 3, n is 8, and z is 2. In some embodiments, x is 5, n is 8, and z is 2. In some embodiments, SM1 is linked to the -S- of a cysteine ​​residue. Various conjugates containing such linkers are described herein.

[0256] In some implementations, the connector-load portion is or includes -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -L T -L RX -L X -(CH2CH2O)nL ZP -LC -R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-L X -(CH2CH2O)nL ZP -L C -R P SM1 is The carbon atom is bonded to the target-binding portion (e.g., an antibody), and each variable is independent as described herein. In some embodiments, the linker-payload portion is or contains -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-L C -R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-L C -R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-GGFG-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-GGFG-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-Val-Ala-PABA-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-Val-Ala-PABA-R PEach variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -C(O)-Val-Cit-PABA-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -C(O)-Val-Cit-PABA-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -N(R')-C(O)-oGlu-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -NH-C(O)-oGlu-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-N(R')-(CH2CH2O)n-(CH2) z -N(R')-C(O)-mGlu-R P Each variable is independent as described herein. In some implementations, the connector-payload portion is or includes -SM1-(CH2). X -C(O)-NH-(CH2CH2O)n-(CH2) z -NH-C(O)-mGlu-R P Each variable is independent as described herein. In some embodiments, x is 2-10. In some embodiments, n is 2-10. In some embodiments, z is 2-10. In some embodiments, x is 2, n is 8, and z is 2. In some embodiments, x is 3, n is 8, and z is 2. In some embodiments, x is 5, n is 8, and z is 2. In some embodiments, R P It is the esxanotecan payload portion. In some implementations, R P yes This document describes various conjugates comprising such linker-load portions. For example, in some embodiments, the target binding portion is or comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the target binding portion is or comprises an anti-HHLA2 antibody or an antigen-binding fragment thereof. In some embodiments, the target binding portion is or comprises Hu2C4-H3L3, 65872, 65889, or 887-1DE or an antigen-binding fragment thereof. In some embodiments, the target binding portion comprises 1, 2, 3, 4, 5, or 6 CDRs of an antibody (e.g., Hu2C4-H3L3, 65872, 65889, or 887-1DE). In some embodiments, the target binding portion is Hu2C4-H3L3, 65872, 65889, or 887-1DE.

[0257] Conjugate In some embodiments, this disclosure provides a conjugate comprising a target-binding portion, a payload portion, and a linker connecting the target-binding portion and the payload portion. In some embodiments, the linker portion comprises a PEG portion. In some embodiments, the linker is or comprises -(CH2CH2O)n-, where n is 1-24. In some embodiments, n is 4. In some embodiments, n is 8. In some embodiments, the -(CH2CH2O)n- in the linker of the conjugate is -(CH2CH2O)4-. In some embodiments, the -(CH2CH2O)n- in the linker of the conjugate is -(CH2CH2O)8-. In some embodiments, the PEG portion in the linker or conjugate is PEG4. In some embodiments, the PEG portion in the linker or conjugate is PEG8. In some embodiments, the payload portion is or comprises a portion of ethatheca or a derivative or analogue thereof, or a pharmaceutically acceptable salt thereof.

[0258] In some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 It is a divisible unit; and L COIt is a covalent bond or a spacer sub-part (e.g., optionally substituted). (where the carbonyl group is bonded to the effective loading portion).

[0259] For example, in some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 Is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; L CO It is a covalent bond or optionally substituted. In this case, the carbonyl group is bonded to the effective load portion; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0260] In some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 Is it -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or ;and L CO It is a covalent bond or optionally substituted. The carbonyl group is bonded to the effective load portion.

[0261] In some embodiments, the linker comprises a PEG moiety. In some embodiments, the linker comprises -(CH2CH2O)n-, where n is 1-24. In some embodiments, the PEG moiety is PEG4. In some embodiments, the PEG moiety is PEG8. This disclosure demonstrates, in particular, that antibody-drug conjugates comprising linkers containing PEG moietyes can provide a variety of benefits and advantages. For example, in some embodiments, using such linkers, the payload portion can be directly linked to a cleavable unit, such as GGFG, without the spacer utilized in the reported products, to provide an antibody-drug conjugate with comparable or improved properties and / or activity. In some embodiments, conjugates having such linkers exhibit reduced aggregation compared to reference conjugates without such linkers. For example, in some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 1-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 It is a divisible unit; and L CO It is a covalent bond or a spacer sub-part (e.g., optionally substituted). (where the carbonyl group is bonded to the effective loading portion).

[0262] For example, in some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 1-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 Is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; L CO It is a covalent bond or optionally substituted. In this case, the carbonyl group is bonded to the effective load portion; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

[0263] In some embodiments, this disclosure provides a conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) The connector that connects the target assembly and the payload assembly; in: The connector contains -(CH2CH2O)n- and L C ; n is 1-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 Is it -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, or ;and L CO It is a covalent bond or optionally substituted. The carbonyl group is bonded to the effective load portion.

[0264] In some embodiments, the conjugate is a compound having the structure of formula A-1 or a salt thereof: , A-1 Each variable is described independently as in this paper.

[0265] In some embodiments, the conjugate is a compound having the structure of formula A-1 or a salt thereof: , A-2 Each variable is described independently as in this paper.

[0266] For example, various implementation schemes of certain variables are described as examples in the section on connectors. Further implementation schemes of the variables are described below as examples.

[0267] R A In some implementations, each R A Independently, it is the target-binding portion as described herein. In some implementations, each R AIndependently, it is a target-binding moiety comprising an antibody or an antigen-binding fragment thereof. In some embodiments, each occurrence of the target-binding moiety is an antibody or an antigen-binding fragment thereof. In some embodiments, each occurrence of the target-binding moiety is an antibody. In some embodiments, each occurrence of the target-binding moiety is a monoclonal antibody. In some embodiments, each occurrence of the target-binding moiety is not trastuzumab. In some embodiments, each occurrence of the target-binding moiety shares less than about 90%, 95%, or 99% sequence identity with trastuzumab. In some embodiments, each occurrence of the target-binding moiety shares less than about 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with trastuzumab. In some embodiments, each occurrence of the target-binding moiety is not an anti-HER2 antibody. In some embodiments, each occurrence of the target-binding moiety is not an anti-HER2 antibody or an antigen-binding fragment thereof.

[0268] In some embodiments, each target-binding moiety of the provided conjugate targets the same agent. In some embodiments, each target-binding moiety targets the same agent, wherein the agent is or comprises a peptide, nucleic acid, or lipid. In some embodiments, each target-binding moiety targets the same agent, wherein the agent is a peptide. In some embodiments, each target-binding moiety targets the same antigen. In some embodiments, each target-binding moiety is independently an antibody or its antigen-binding fragment, and each target-binding moiety shares about 90%, 95%, 96%, 97%, 98%, or 99% sequence identity. In some embodiments, each target-binding moiety targets the same antigen. In some embodiments, each target-binding moiety is independently an antibody or its antigen-binding fragment, and each target-binding moiety shares the same amino acid sequence. In some embodiments, each target-binding moiety is identical.

[0269] In some embodiments, the target binding moiety is or comprises an antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is or comprises an anti-HHLA2 antibody or its antigen-binding fragment. In some embodiments, the target binding moiety is or comprises Hu2C4-H3L3, 65872, 65889, or 887-1DE or its antigen-binding fragment. In some embodiments, the target binding moiety comprises 1, 2, 3, 4, 5, or 6 CDRs of an antibody (e.g., Hu2C4-H3L3, 65872, 65889, or 887-1DE). In some embodiments, the target binding moiety is Hu2C4-H3L3, 65872, 65889, or 887-1DE.

[0270] s In some embodiments, the conjugate contains a single target-binding moiety. In some embodiments, the conjugate contains two or more target-binding moieties.

[0271] In some implementations, s is 1. In some implementations, s is 2-20, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20.

[0272] a In some embodiments, the connector is bonded to a single site on the target binding portion. In some embodiments, the occurrence of 'a' is 1. In some embodiments, each occurrence of 'a' is 1.

[0273] In some embodiments, the linker is bonded to two or more target-binding moieties. In some embodiments, the linker bond is bonded to two or more sites on the target-binding moieties (e.g., two or more Cys residues of an antibody or its antigen-binding fragment). In some embodiments, a is greater than 1. In some embodiments, the occurrence of a is 2-10, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10. In some embodiments, the occurrence of a is 2. In some embodiments, the occurrence of a is 3-10.

[0274] In some implementations, each 'a' is the same.

[0275] For each In some implementations, each L T They are the same; in some implementations, each L RX They are the same; in some implementations, each -L T -L RX - They are the same.

[0276] In some implementations, each L T They are the same. In some implementations, each L RX They are the same. In some implementations, each -L T -L RX - They are the same.

[0277] b In some implementations, the connector is bonded to a single site on the payload portion. In some implementations, the occurrence of b is 1. In some implementations, each occurrence of b is 1.

[0278] In some embodiments, the connector is bonded to two or more payload portions. In some embodiments, the connector bond is bonded to two or more sites on the payload portion. In some embodiments, b is greater than 1. In some embodiments, b occurs in the range of 2-10, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10. In some embodiments, b occurs in the range of 2. In some embodiments, b occurs in the range of 3-10.

[0279] In some implementations, b is the same.

[0280] In some implementations, each L X 'They are the same.'

[0281] In some implementations, the at least one occurrence of n is independently 1-24. In some implementations, each occurrence of n is independently 1-24. In some implementations, each n is 8. For each In some implementations, each n is the same; in some implementations, each L ZP They are the same; in some implementations, each L C They are the same; in some implementations, each R P They are the same; in some implementations, each -(CH2CH2O)nL ZP - are the same; in some implementations, each -(CH2CH2O)nL ZP -L C - are the same; in some implementations, each -(CH2CH2O)nL ZP -L C -R P They are the same; in some implementations, each -L C -R P They are the same; in some implementations, -L ZP -L C -R P They are the same.

[0282] In some implementations, each n is the same. In some implementations, each L... ZP They are the same. In some implementations, each L C They are the same. In some implementations, each R P They are the same. In some implementations, each -(CH2CH2O)nL ZP - They are the same. In some implementations, each -(CH2CH2O)nL ZP -L C - They are the same. In some implementations, each -(CH2CH2O)nLZP -L C -R P They are the same. In some implementations, each -L C -R P They are the same. In some implementations, -L ZP -L C -R P They are the same.

[0283] t In some embodiments, the conjugate contains a single joint portion. In some embodiments, the conjugate contains two or more joint portions.

[0284] In some implementations, t is 1. In some implementations, t is 2-20 (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 2-15, 2-10, 2-8, etc.). In some implementations, t is about 1-8. In some implementations, t is about 2-8. In some implementations, t is between about 1 and 16. In some implementations, t is between about 1 and 12. In some implementations, t is between about 2 and 10. In some implementations, t is between about 4 and 8. In some implementations, t is about 1. In some implementations, t is about 2. In some implementations, t is about 3. In some implementations, t is about 4. In some implementations, t is about 5. In some implementations, t is about 6. In some implementations, t is about 7. In some implementations, t is about 8. In some implementations, t is an integer between about 7 and about 9. In some implementations, t is an integer between approximately 6 and approximately 10.

[0285] In some embodiments, s is 1, a is 1, and b is 1, and t is a DAR as described herein. In some embodiments, for a conjugate having formula A-2 or a salt thereof, t is a DAR as described herein.

[0286] For example, in some implementations, L X Each occurrence of C is independently a covalent bond or optionally substituted divalent linear or branched C. 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L X Each occurrence of is -(CH2) X-C(O)-N(R')-, where x is 1-19, and -N(R')- is bonded to -(CH2CH2O)n-. In some implementations, L X Each occurrence of is -(CH2) X -C(O)-NH-, where x is 1-19, and -NH- is bonded to -(CH2CH2O)n-. In some embodiments, L X Each occurrence of is -(CH2) X -C(O)-N(R')- or -(CH2) X -C(O)-NH-, and x is 1-6. In some implementations, L X Each occurrence is -(CH2)5-C(O)-N(R')-, where -N(R')- is bonded to -(CH2CH2O)n-. In some implementations, L X Each occurrence of -(CH2)5-C(O)-NH-, where -NH- is bonded to -(CH2CH2O)n-. In some embodiments, L X Each occurrence is -(CH2)3-C(O)-N(R')-, where -N(R')- is bonded to -(CH2CH2O)n-. In some implementations, L X Each occurrence of L is -(CH2)3-C(O)-NH-, where -NH- is bonded to -(CH2CH2O)n-. In some embodiments, L X The same thing happens every time it appears.

[0287] In some implementations, L T The presence of L is a covalent bond. In some implementations, L T The emergence of C is the optional replacement of divalent straight-chain or branched C. 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, and -S(O)2(NR')-. In some embodiments, L T The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic group. In some embodiments, L T The appearance of C is a binary straight chain or branch chain. 1-20 Aliphatic group. In some embodiments, L T The appearance of -(CH2) is optional and is replaced. 1-5 - In some implementations, L T The appearance of -(CH2) 1-5- In some implementations, L T The presence of -CH2-. In some implementations, L T The presence of L is associated with the backbone atoms of amino acid residues in the target-binding region. In some embodiments, L T The presence of L is independently bonded to the backbone atoms of amino acid residues in the same target-binding moiety. In some embodiments, L T The presence of L is due to the bonding of different backbone atoms of amino acid residues in the same target-binding region. In some embodiments, L T The presence of L is associated with the α-atom bonding of amino acid residues in the target-binding moiety. In some embodiments, L T The presence of L is associated with the α-atom bonding of amino acid residues in the same target-binding moiety. In some embodiments, L T The presence of L is associated with different α-atoms bonded to amino acid residues in the same target-binding moiety. In some embodiments, L T The same thing happens every time it appears.

[0288] In some implementations, L RX The presence of L is a covalent bond. In some implementations, L RX The appearance is due to the presence of groups on the unattached target-binding site and the presence of -L X -(CH2CH2O)nL ZP -L C - The portion produced by the reaction between the active loading portion and the groups present on the agent. In some embodiments, L RX The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-, wherein each -Cy- is independently a 3-12 membered ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

[0289] In some implementations, L RX The appearance of -L RX1 -L RX2 -, where L RX1 and L RX2 Each of them is independent as described in this article.

[0290] In some implementations, -L RX1 The presence of - is -S-. In some implementations, -L RX1The presence of - is -S-, and -S- is a Cys residue of the target-binding moiety. In some embodiments, -L RX1 The presence of - is -S-, and -S- is a Cys residue in the antibody, wherein the Cys residue forms an interchain disulfide bond in the unconjugated antibody. In some embodiments, -L RX1 The presence of - is -S-, and -S- is a Cys residue in the antibody, where the Cys residue forms a disulfide bond between the heavy and light chains in the unconjugated antibody. In some embodiments, -L RX1 The presence of - is -S-, and -S- is of the target-binding Cys residue, wherein the Cys residue is derived by partial or complete reduction of interchain disulfide bonds.

[0291] In some implementations, -L RX2 - The appearance of - is Nitrogen and L X Bonded, and carbon with L RX1 Bonding. In some implementations, -L RX2 The appearance of - is -S-. In some implementations, L RX The presence of L is or includes a click chemical product portion. In some embodiments, L RX It may contain a click chemical product portion. In some embodiments, L RX The appearance of is or contains In some implementations, where L RX The appearance of is or contains In some implementations, L RX The appearance of is or contains In some implementations, the nitrogen in the 5-membered ring reacts with L... X Bonding. In some implementations, the carbon or nitrogen in the 8-membered ring bonds with L. X Bonding.

[0292] In some implementations, n occurs in the range of 1-24. In some implementations, n occurs in the range of 4-24. In some implementations, n occurs in the range of 7-20. In some implementations, each occurrence of n is independently 4-24. In some implementations, each occurrence of n is independently 7-20. In some implementations, n occurs in the range of 8. In some implementations, each n is the same. In some implementations, each n is 8.

[0293] In some implementations, L ZP The presence of L is a covalent bond. In some implementations, L ZP The emergence of C is the optional replacement of divalent straight-chain or branched C. 1-20 Aliphatic, of which L ZThe 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')-, and -Cy-. In some embodiments, L ZP The form is -(CH2)zC(O)-, where z is 1-19. In some implementations, L ZP The appearance is -(CH2)2-C(O)-, where -C(O)- and -L C - The amino bond of the first amino acid in -. In some embodiments, each L ZP They are the same.

[0294] R P As used in this article, each R P Independently, it is a payload portion. Various payload portions are available and can be utilized according to this disclosure. In some embodiments, each R... P They are the same. In some implementations, R P The presence of [a substance] is due to its cytotoxicity. In some implementations, each R [substance]... P It is an independent cytotoxic agent. In some implementations, R P The presence of [the substance] is due to esanotecan or its derivatives. In some embodiments, each R [contains / contains / etc.]. P It is independently esanotecan or a derivative thereof. In some embodiments, R P The appearance of is ("R" P -Ex" or "R" EX In some implementations, each R P It is R P -Ex. In some implementations, R P The appearance of is In some implementations, R P Each time it appears is .

[0295] In some implementations, the conjugate contains L C -R P In some implementations, -L C -R P The appearance of -GGFG-R P In some implementations, it is -Val-Ala-PABA-R P In some implementations, it is -Val-Cit-PABA-R P In some implementations, it is -Val-Ala-NH-oGlu-RP In some implementations, it is -Val-Ala-NH-mGlu-R P In some implementations, it is -Val-Cit-NH-oGLu-R P In some implementations, it is -Val-Cit-NH-mGLu-R P In some implementations, it is -oGlu-R P In some implementations, it is -mGlu-R P In some implementations, R P It contains a nitrogen atom and is bonded at the nitrogen atom, for example, to the rest of the conjugate or other compound. In some embodiments, R P It is R EX In some implementations, -L C -R P The same thing happens every time it appears.

[0296] In some implementations, -L C -R P The appearance of is , , , , , or In some implementations, each -L C -R P They are the same.

[0297] In some implementations, -L C -R P The appearance of is or In some implementations, each -L C -R P They are the same.

[0298] In some implementations, -L C -R P The appearance of is .

[0299] In some implementations, -L C -R P Each appearance is independent .

[0300] In some implementations, -L C -R P The appearance of is .

[0301] In some implementations, -L C -R P Each appearance is .

[0302] In some implementations, -L C -R P The appearance of is .

[0303] In some implementations, -L C -R P Each appearance is .

[0304] In some implementations, -L C -R P The appearance of is In some implementations, -L C -R P Each appearance is In some implementations, -L C -R P The appearance of is In some implementations, -L C -R P Each time it appears is In some implementations, -L C -R P The appearance of is In some implementations, -L C -R P Each appearance is In some implementations, -L C -R P The appearance of is In some implementations, -L C -R P Each appearance is .

[0305] In some implementations, the conjugate contains -L ZP -L C -R P In some implementations, -L ZP -L C -R P The appearance of -(CH2) z C(O)-GGFG-R P In some implementations, it is -(CH2). z C(O)-Val-Ala-PABA-R PIn some implementations, it is -(CH2). z C(O)-Val-Cit-PABA-R P In some implementations, it is -(CH2). z C(O)-Val-Ala-NH-oGlu-R P In some implementations, it is -(CH2). z C(O)-Val-Ala-NH-mGlu-R P In some implementations, it is -(CH2). z C(O)-Val-Cit-NH-oGLu-R P In some implementations, it is -(CH2). z C(O)-Val-Cit-NH-mGLu-R P In some implementations, it is -(CH2). z NHC(O)-oGlu-R P In some implementations, it is -(CH2). z NHC(O)-mGlu-R P In some implementations, z is 2. In some implementations, R P It contains a nitrogen atom and is bonded at the nitrogen atom, for example, to the rest of the conjugate or other compound. In some embodiments, R P It is R EX In some implementations, z is 2, and R P It is R EX In some implementations, -L ZP -L C -R P The same thing happens every time it appears.

[0306] In some implementations, -L ZP -L C -R P The appearance of is , , , , , , , or In some implementations, the occurrence is the same each time.

[0307] In some implementations, -L ZP -L C -R P The appearance of is or In some implementations, the occurrence is the same each time.

[0308] In some implementations, -L ZP -L C -R P The appearance of is or contains .

[0309] In some implementations, -L ZP -L C -R P Each occurrence of is or contains .

[0310] In some implementations, -L ZP -L C -R P The appearance of is or contains .

[0311] In some implementations, -L ZP -L C -R P Each occurrence of is or contains .

[0312] In some implementations, -L ZP -L C -R P The appearance of is or contains .

[0313] In some implementations, -L ZP -L C -R P Each occurrence of is or contains .

[0314] In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains .

[0315] In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -LZP -L C -R P Each occurrence of is or contains In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains In some implementations, -L ZP -L C -R P The appearance of is or contains In some implementations, -L ZP -L C -R P Each occurrence of is or contains .

[0316] In some embodiments, the conjugate contains -(CH2CH2O)nL ZP -L C -R P In some implementations, -(CH2CH2O)nL ZP -L C -R PThe appearance is -(CH2CH2O)n-(CH2) z C(O)-GGFG-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)-Val-Ala-PABA-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)-Val-Cit-PABA-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)Val-Ala-NH-oGlu-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)-Val-Ala-NH-mGlu-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)-Val-Cit-NH-oGLu-R P In some implementations, it is -(CH2CH2O)n-(CH2). z C(O)-Val-Cit-NH-mGLu-R P In some implementations, it is -(CH2CH2O)n-(CH2). z NHC(O)-oGlu-R P In some implementations, it is -(CH2CH2O)n-(CH2). z NHC(O)-mGlu-R P In some implementations, n is 8. In some implementations, z is 2. In some implementations, R P It contains a nitrogen atom and is bonded at the nitrogen atom, for example, to the rest of the conjugate or other compound. In some embodiments, R P It is R EX In some implementations, z is 2, and R P It is R EX In some implementations, n is 8, z is 2, and R P It is R EX In some implementations, -(CH2CH2O)nL ZP -L C -R P The same thing happens every time it appears.

[0317] In some implementations, -(CH2CH2O)nL ZP-L C -R P The appearance of is or contains , , , , , , , or In some implementations, the occurrence is the same each time.

[0318] In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains or . In some implementations, the occurrence is the same each time.

[0319] In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0320] In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0321] In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0322] In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0323] In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0324] In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0325] In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R PEach occurrence of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0326] In some implementations, the conjugate contains -L X -(CH2CH2O)nL ZP -L C -R P In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of -(CH2) x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-GGFG-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Ala-PABA-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-PABA-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)Val-Ala-NH-oGlu-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Ala-NH-mGlu-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-NH-oGLu-R P In some implementations, it is -(CH2).x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-NH-mGLu-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z NHC(O)-oGlu-R P In some implementations, it is -(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z NHC(O)-mGlu-R P In some implementations, x is 2. In some implementations, x is 3. In some implementations, x is 5. In some implementations, n is 8. In some implementations, z is 2. In some implementations, R P It contains a nitrogen atom and is bonded at the nitrogen atom, for example, to the rest of the conjugate or other compound. In some embodiments, R P It is R EX In some implementations, z is 2, and R P It is R EX In some implementations, n is 8, z is 2, and R P It is R EX In some implementations, x is 2, n is 8, z is 2, and R... P It is R EX In some implementations, x is 3, n is 8, z is 2, and R... P It is R EX In some implementations, x is 5, n is 8, z is 2, and R... P It is R EX In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The same thing happens every time it appears.

[0327] , , , , , , , or In some implementations, the occurrence is the same each time.

[0328] In some implementations, -L X-(CH2CH2O)nL ZP -L C -R P The appearance of is or contains or In some implementations, the occurrence is the same each time.

[0329] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0330] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0331] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0332] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0333] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0334] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0335] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R PEach occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0336] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains , , , , , , , or In some implementations, the occurrence is the same each time.

[0337] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains or In some implementations, the occurrence is the same each time.

[0338] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0339] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0340] In some implementations, -L X-(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0341] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0342] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0343] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0344] In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -LC -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0345] In some implementations, the conjugate contains -L RX -L X -(CH2CH2O)nL ZP -L C -R P In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of -S-SM-(CH2)x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-GGFG-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Ala-PABA-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-PABA-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)Val-Ala-NH-oGlu-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Ala-NH-mGlu-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-NH-oGLu-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z C(O)-Val-Cit-NH-mGLu-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z NHC(O)-oGlu-R P In some implementations, it is -S-SM-(CH2). x C(O)NH-(CH2CH2O)n-(CH2) z NHC(O)-mGlu-R P In some implementations, x is 2. In some implementations, x is 3. In some implementations, x is 5. In some implementations, n is 8. In some implementations, z is 2. In some implementations, R PIt contains a nitrogen atom and is bonded at the nitrogen atom, for example, to the rest of the conjugate or other compound. In some embodiments, R P It is R EX In some implementations, z is 2, and R P It is R EX In some implementations, n is 8, z is 2, and R P It is R EX In some implementations, x is 2, n is 8, z is 2, and R... P It is R EX In some implementations, x is 3, n is 8, z is 2, and R... P It is R EX In some implementations, x is 5, n is 8, z is 2, and R... P It is R EX In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The same thing happens every time it appears.

[0346] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains , , , , , , , or In some implementations, the occurrence is the same each time.

[0347] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains or In some implementations, the occurrence is the same each time.

[0348] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -LC -R P The appearance of is or contains .

[0349] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0350] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0351] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0352] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains .

[0353] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0354] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -LRX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C-R P Each occurrence of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains .

[0355] In some implementations, -L RX -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains , , , , , , , or In some implementations, the occurrence is the same each time. ...

Claims

1. A conjugate comprising: i) Targeting the target-binding region of HHLA2; ii) The payload portion having the following structure: ;as well as iii) A connector that connects the target assembly portion and the payload portion; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C yes , , or The carbonyl group is bonded to the effective load portion.

2. The conjugate of claim 1, wherein the targeting binding portion is an antibody selected from Hu2C4-H3L3, 65872, 65889 or 887-1DE.

3. The conjugate as claimed in claim 2, wherein n is 1-24.

4. The conjugate as claimed in claim 3, wherein L C yes , or .

5. The conjugate as claimed in claim 2, wherein n is 0, and L C yes .

6. A conjugate comprising: i) Target-binding portion; ii) The payload portion; and iii) A connector that connects the target assembly portion and the payload portion; in: The connector contains -(CH2CH2O)n- and L C ; n is 0-24; L C Partially bonded to the payload and is -L C1 -L CO -; L C1 It is a divisible unit; or L C1 Is -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; L CO It is a covalent bond or a spacer sub-part (e.g., optionally substituted). (where the carbonyl group is partially bonded to the effective load); Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

7. The conjugate of claim 6, wherein the target-binding portion comprises an antibody or an antigen-binding fragment thereof.

8. The conjugate as claimed in claim 6 or 7, wherein the effective loading portion is 。 9. The conjugate as claimed in any of the preceding claims, wherein the ratio (DAR) of the effective load portion to the target binding portion of the conjugate is about 2-8.

10. The conjugate as claimed in any of the preceding claims, wherein the connector comprises -(CH2CH2O)nL ZP -L C -,in: L ZP It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each -Cy- is independently a divalent 3- to 20-membered ring having 0-10 independently substituted heteroatoms selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and R is C, which can be arbitrarily replaced. 1-6 Lipids.

11. The conjugate as claimed in any of the preceding claims, wherein the connector comprises -L X -(CH2CH2O)nL ZP -L C -,in: L X It is a covalent bond or an optionally substituted divalent or multivalent straight or branched chain C. 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L ZP It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each -Cy- is independently a 3-12 membered ring having 0-4 independently substituted heteroatoms selected from nitrogen, oxygen, and sulfur; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

12. The conjugate as claimed in any of the preceding claims, wherein the connector is or comprises -L RX -L X -(CH2CH2O)nL ZP -L C -,in: L RX It is a covalent bond or a group present on an unattached target-binding site and containing -L X -(CH2CH2O)nL ZP -L C - The part produced by the reaction between - and the groups present on the agent of the effective loading portion, wherein -L C - Bonded to the payload portion; or L RX It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L X It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L ZP It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each -Cy- is independently a 3-12 membered ring having 0-4 independently substituted heteroatoms selected from nitrogen, oxygen, and sulfur; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

13. The conjugate as claimed in any of the preceding claims, wherein the connector comprises -L T -L RX -L X -(CH2CH2O)nL ZP -L C -,in: L T It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L RX It is a covalent bond or a group present on an unattached target-binding site and containing -L X -(CH2CH2O)nL ZP -L C - The part produced by the reaction between - and the groups present on the agent of the effective loading portion, wherein -L C - Bonded to the payload portion; or L RX It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L X It is a covalent bond or an optionally substituted divalent or multivalent straight or branched chain C. 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; L ZP It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each -Cy- is independently a 3-12 membered ring having 0-4 independently substituted heteroatoms selected from nitrogen, oxygen, and sulfur; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

14. The conjugate according to any one of claims 10 to 13, wherein L ZP It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L Z The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')- and -S(O)2(NR')-.

15. The conjugate of claim 14, wherein L ZP It is -(CH2)zN(R')C(O)-, -(CH2)zC(O)- or -(CH2)zC(O)N(R')-, where z is 1-19.

16. The conjugate according to any one of claims 6 to 15, wherein L C is -Gly-Gly-Phe-Gly-, -Val-Ala-PABA-, -Val-Cit-PABA-, mGlu or oGlu.

17. The conjugate as claimed in any of the preceding claims, wherein n is 8.

18. The conjugate according to any one of claims 11 to 17, wherein L X It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-.

19. The conjugate as claimed in any one of claims 12 to 18, wherein each L RX Independently is -L RX1 -L RX2 -,in: Each L RX1 Independently covalent or optionally substituted divalent or multivalent straight or branched C 1-10 Aliphatic, of which L RX1 The 1-5 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX2 Independently covalent or optionally substituted divalent straight or branched C 1-10 Aliphatic, of which L RX2 The 1-5 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each -Cy- is independently a 3- to 12-membered ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen and sulfur, which are optionally substituted. Each L RX2 With L X Bonding; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

20. The conjugate according to any one of claims 13 to 19, wherein L T It is a covalent bond or an optionally substituted divalent straight chain or branch C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')- and -S(O)2(NR')-.

21. The conjugate according to any one of claims 13 to 20, wherein L T It is -CH2-.

22. The conjugate of any one of claims 13 to 21, wherein the target-binding moiety comprises an amino acid residue, and L T It is bonded to the main chain atoms of the amino acid residues.

23. The conjugate according to any one of claims 6 to 10, wherein the connector is or comprises ,in: Each L RX Independently covalently bonded or formed by groups present on the unattached target-binding moiety and containing L X’ The portion produced by the reaction between the agent and the optional effective loading portion and the groups present on the agent, wherein -L C - Bonded to the payload portion; or each L RX Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each of a and b is independently 1-10; L X’ It is a covalent bond or an optionally substituted divalent or multivalent straight or branched chain C. 1-30 Aliphatic, of which L X’ The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; Each L C1S Independently In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; Each L CO Not existing independently or being arbitrarily replaced In this case, the carbonyl group is bonded to the effective load portion; Each L C Independently bonded to the payload portion; Each -Cy- is independently a 3- to 12-membered ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen and sulfur, which are optionally substituted. Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

24. The conjugate according to any one of claims 6 to 10, wherein the connector is or comprises ,in: Each L T Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX Independently covalently bonded or formed by groups present on the unattached target-binding moiety and containing L X’ The portion produced by the reaction between the agent and the optional effective loading portion and the groups present on the agent, wherein -L C - Bonded to the payload portion; or each L RX Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each of a and b is independently 1-10; L X’ It is a covalent bond or an optionally substituted divalent or multivalent straight or branched chain C. 1-30 Aliphatic, of which L X’ The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; Each L C1S Independently In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; Each L CO Not existing independently or being arbitrarily replaced In this case, the carbonyl group is bonded to the effective load portion; Each L C Independently bonded to the payload portion; Each -Cy- is independently a 3- to 12-membered ring having 0-4 independently selected heteroatoms chosen from nitrogen, oxygen and sulfur, which are optionally substituted. Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or optionally substituted C. 1-6 Lipids.

25. The conjugate of claim 24, wherein each L T Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')- and -S(O)2(NR')-.

26. The conjugate according to any one of claims 24 to 25, wherein L T Each occurrence of it is -CH2-.

27. The conjugate as claimed in any of the preceding claims, wherein the connector is or comprises a structure of formula B: ， B in: # indicates the connection point with the target; % indicates the connection point with the payload section; X is a portion generated by the reaction of unbonded connectors and groups present on the target-bonded portion; Y is L C ;and Z is or contains -(CH2CH2O)n-.

28. A conjugate, wherein the conjugate is a compound of formula A-1 or a salt thereof: ， A-1 in: Each R A It is a target-binding component independently; s is 1-20; Each L T Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX Independently covalent bonds, consisting of groups present on the unattached target-binding portion and containing L X’ The reaction between groups present on the agent produces partially or optionally substituted divalent straight-chain or branched C atoms. 1-20 Aliphatic, of which L RX 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; or each L RX Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each of a and b is independently 1-10; Each L X’ Independently covalent or optionally substituted divalent or multivalent straight or branched C 1-30 Aliphatic, of which L X’ The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 Independently, it is a divisible unit, or each L C1 Independently, they are -Gly-Gly-Phe-Gly-, -Val-Ala-, -Val-Cit-, L C1S -Gly-Gly-Phe-Gly-N(R')-L C1S -、-Val-Ala-N(R')-L C1S -or-Val-Cit-N(R')-L C1S -; L C1S yes In which the benzene ring and -CH2- are optionally substituted, and the carbonyl group is associated with L CO Bonding, and R LS It may contain a sugar component; Each L CO Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L CO The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each R P It is a separate payload component; t is 1-20; Each -Cy- is independently a divalent 3- to 20-membered ring having 0-10 independently substituted heteroatoms selected from silicon, nitrogen, oxygen, phosphorus and sulfur; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

29. The conjugate of claim 28, wherein s is 1.

30. The conjugate according to any one of claims 28 to 29, wherein the appearance of a is 2-10.

31. The conjugate according to any one of claims 28 to 30, wherein the appearance of b is 2-10.

32. A conjugate, wherein the conjugate is a compound of formula A-2 or a salt thereof: ， A-2 in: R A It is the target-binding part; Each L T Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L RX Independently covalent bonds, consisting of groups present on the unattached target-binding moiety and containing -L X -(CH2CH2O)nL ZP -L C -R P The reaction between groups present on the agent produces partially or optionally substituted divalent straight-chain or branched C atoms. 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Or each L X Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each n is independently 0-24; Each L ZP Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L ZP The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each L C Independently is -L C1 -L CO -; Each L C1 It is an independent, divisible unit; Each L CO Independently covalent or optionally substituted divalent straight or branched C 1-20 Aliphatic, of which L CO The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-; Each R P It is a separate payload component; t is 1-20; Each -Cy- is independently a divalent 3- to 20-membered ring having 0-10 independently substituted heteroatoms selected from silicon, nitrogen, oxygen, phosphorus and sulfur; Each R' is independently -R, -C(O)R, -CO2R, or -S(O)2R, or two or more R's together with their intermediate atoms form optionally substituted 3-20-membered rings, which, in addition to the intermediate atoms, also have 0-10 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur; and Each R is independently hydrogen or a group selected from the following optionally substituted groups: C 1-10 Aliphatic; having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus, and sulfur. 1-10 Mixed lipids; C 6-14 Aryl; a 5-14 membered heteroaryl ring having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur; and a 3-15 membered heterocycle having 1-5 heteroatoms independently selected from silicon, nitrogen, oxygen, phosphorus and sulfur.

33. The conjugate of claim 32, wherein R A It may contain antibodies or their antigen-binding fragments.

34. The conjugate according to any one of claims 32 to 33, wherein L X The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic, of which L X The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-.

35. The conjugate according to any one of claims 28 to 34, wherein L T The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic, of which L T The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-.

36. The conjugate according to any one of claims 28 to 35, wherein L T The appearance of -CH2-.

37. The conjugate according to any one of claims 28 to 36, wherein L RX The appearance of is either a covalent bond or a group present on the unattached target-binding site and a group containing -L X -(CH2CH2O)nL ZP -L C - The part produced by the reaction between the effective loading portion and the groups present on the agent, or where L RX The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic, of which L RX The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-, wherein each -Cy- is independently a 3-12 membered ring having 0-4 heteroatoms independently selected from nitrogen, oxygen and sulfur.

38. The conjugate according to any one of claims 28 to 37, wherein the occurrence of n is 8.

39. The conjugate according to any one of claims 28 to 38, wherein L ZP The appearance of C is a covalent bond or an optionally replaced divalent straight chain or branch C. 1-20 Aliphatic, of which L Z The 1-10 methylene units are optionally and independently replaced by groups selected from -O-, -N(R')-, -C(O)-, -S-, -S(O)-, -S(O)2-, -C(O)N(R')-, -S(O)2(NR')- and -Cy-.

40. The conjugate according to any one of claims 28 to 39, wherein each R P yes 。 41. The conjugate according to any one of claims 28 to 33, wherein each -L C -R P Independently , , or .

42. The conjugate according to any one of claims 28 to 33, wherein -L ZP -L C -R P Each occurrence of is or contains , , or .

43. The conjugate according to any one of claims 28 to 33, wherein -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains , , or .

44. The conjugate according to any one of claims 28 to 33, wherein -L X -(CH2CH2O)nL ZP -L C -R P The appearance of is or contains , , or .

45. The conjugate according to any one of claims 28 to 33, wherein -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains , , or .

46. ​​The conjugate according to any one of claims 28 to 33, wherein -L T -L RX -L X -(CH2CH2O)nL ZP -L C -R P Each occurrence of is or contains , , or .

47. The conjugate of any one of claims 28 to 46, wherein the target-binding moiety comprises an amino acid residue, and -L T -L RX -L X -(CH2CH2O)nL ZP -L C -R P It is bonded to the main chain atoms of the amino acid residues.

48. The conjugate of claim 6, having a structure of formula XC: XC Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

49. The conjugate as claimed in claim 6, having the structure of formula X-C': X-C' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

50. The conjugate as claimed in claim 6, having the structure of formula X-G': X-G' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

51. The conjugate as claimed in claim 6, having the structure of formula X-H': X-H' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

52. The conjugate as claimed in claim 6, having the structure of formula X-I': X-I' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

53. The conjugate as claimed in claim 6, having the structure of formula X-J': X-J' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

54. The conjugate as claimed in claim 6, having the structure of formula X-K': X-K' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

55. The conjugate as claimed in claim 6, having the structure of formula X-L': X-L' Or the structure of a pharmaceutically acceptable salt thereof, wherein: R A It is the target-binding part; and t is an integer between 1 and 16.

56. The conjugate according to any one of claims 28 to 55, wherein t is about 2-8.

57. The conjugate as claimed in any of the preceding claims, wherein each target-binding moiety is independently an antibody or an antigen-binding fragment thereof.

58. The conjugate according to any one of claims 6 to 56, wherein each target-binding moiety is an anti-HHLA2 antibody or an antigen-binding fragment thereof.

59. The conjugate of claim 58, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: (a) Heavy chain variable regions (VHs) comprising one, two, or three VH CDR sequences each having at least approximately 90% identity with the VH CDRs in Table 1; and / or (b) Light chain variable region (VL) which contains one, two or three VL CDR sequences that each have at least about 90% identity with the VL CDRs in Table 1.

60. The conjugate of claim 58, wherein the anti-HHLA2 or its antigen-binding fragment is or comprises: (a) VH, which includes or consists of the VH in Table 1; and / or (b) VL, which includes or consists of the VL in Table 1.

61. The conjugate of any one of claims 58 to 60, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: Heavy chain variable region (VH) containing a VHCDR sequence that has about or at least about 90% identity with HCDR1 of 887-1DE in Table 1; Heavy chain variable region (VH) containing a VHCDR sequence that has about or at least about 90% identity with HCDR2 of 887-1DE in Table 1; Heavy chain variable region (VH) containing a VHCDR sequence that has about or at least about 90% identity with HCDR3 of 887-1DE in Table 1; The light chain variable region (VL) contains a VLCDR sequence that has about or at least about 90% identity with the LCDR1 of 887-1DE in Table 1; Light chain variable region (VL) containing a VLCDR sequence having approximately or at least approximately 90% identity with LCDR2 of 887-1DE in Table 1; and / or The light chain variable region (VL) contains a VLCDR sequence that has about or at least about 90% identity with LCDR3 of 887-1DE in Table 1.

62. The conjugate of any one of claims 58 to 60, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR1 of 65889 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR2 of 65889 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR3 of 65889 in Table 1; Light chain variable region (VL) containing a VL CDR sequence that has about or at least about 90% identity with LCDR1 of 65889 in Table 1; Light chain variable region (VL) containing a VL CDR sequence having approximately or at least approximately 90% identity with LCDR2 of 65889 in Table 1; and / or Light chain variable region (VL) containing a VL CDR sequence that has about or at least about 90% identity with LCDR3 of 65889 in Table 1.

63. The conjugate of any one of claims 58 to 60, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR1 of 65872 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR2 of 65872 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR3 of 65872 in Table 1; Light chain variable region (VL) containing a VL CDR sequence that has about or at least about 90% identity with LCDR1 of 65872 in Table 1; Light chain variable region (VL) containing a VL CDR sequence having approximately or at least approximately 90% identity with LCDR2 of 65872 in Table 1; and / or The light chain variable region (VL) contains a VL CDR sequence that has about or at least about 90% identity with LCDR3 of 65872 in Table 1.

64. The conjugate of any one of claims 58 to 60, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises: Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR1 of Hu2C4-H3L3 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with HCDR2 of Hu2C4-H3L3 in Table 1; Heavy chain variable region (VH) containing a VH CDR sequence that has about or at least about 90% identity with the HCDR3 of Hu2C4-H3L3 in Table 1; Light chain variable region (VL) containing a VL CDR sequence that has about or at least about 90% identity with the LCDR1 of Hu2C4-H3L3 in Table 1; Light chain variable region (VL) containing a VL CDR sequence having approximately or at least approximately 90% identity with the LCDR2 of Hu2C4-H3L3 in Table 1; and / or The light chain variable region (VL) contains a VL CDR sequence that has about or at least about 90% identity with the LCDR3 of Hu2C4-H3L3 in Table 1.

65. The conjugate according to any one of claims 58 to 60, wherein the anti-HHLA2 antibody or its antigen-binding fragment is or comprises 887-1DE, 65889, 65872 or Hu2C4H3L3 in Table 1.

66. The conjugate according to any one of claims 58 to 60, wherein the K+ is about 15 nM or less. D Combined with HHLA2.

67. A conjugate, wherein the conjugate is a compound having the following structure: 、 or Or salts of these compounds, where R A It is 887-1DE, 65889, 65872 or Hu2C4H3L3 in Table 1, each -S- is independently a Cys residue, and t is about 1-8.

68. The conjugate of claim 67, wherein t is about 2-8.

69. The conjugate as claimed in any of the preceding claims, wherein the conjugate is in a pharmaceutically acceptable salt form.

70. A composition comprising one or more conjugates, each of the conjugates being independently a conjugate as described in any of the preceding claims.

71. The composition of claim 70, wherein the ratio of the effective load portion to the target binding portion of the composition is about 4-8.

72. The composition of any one of claims 70 to 71, wherein the composition is a pharmaceutical composition.

73. The composition of any one of claims 70 to 72, wherein the composition comprises a pharmaceutically acceptable carrier.

74. A kit comprising a conjugate as described in any one of claims 1 to 69 or a composition as described in any one of claims 70 to 73, and optionally instructions for use and / or application.

75. A method for modulating the immune response of a subject, comprising administering or delivering to a subject in need a therapeutically effective amount of the conjugate or composition as described in any one of claims 1 to 73.

76. A method of treating a subject suffering from a disease, symptom, or condition, comprising administering or delivering to the subject in need a therapeutically effective amount of the conjugate or composition as described in any one of claims 1 to 73.

77. The method of any one of claims 75 to 76, wherein the subject has cancer.

78. The method of any one of claims 75 to 77, wherein the subject has a solid tumor, or wherein the subject has kidney cancer, bone cancer, skin cancer, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, lung cancer, ovarian cancer, liver cancer, prostate cancer, pancreatic cancer, bile duct cancer, or thyroid cancer, or wherein the subject has a hematologic malignancy, or wherein the subject has leukemia, or wherein the subject has acute lymphoblastic leukemia, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, chronic leukemia, or acute leukemia, or wherein the subject has lymphoma, or wherein the subject has Hodgkin lymphoma (HL), non-Hodgkin lymphoma, lymphocytic lymphoma, or diffuse large B-cell lymphoma (DLBCL), or wherein the subject is at risk of developing cancer.

79. The method of any one of claims 76 to 78, wherein the disease, symptom, or condition is associated with abnormal HHLA2 expression.

80. The method of any one of claims 75 to 79, wherein the conjugate is applied in combination with the second agent.

81. A method for inhibiting the proliferation of diseased cells in a system, comprising administering or delivering to the system a conjugate or composition as described in any one of claims 1 to 73, wherein the diseased cells express or contain an agent to which a target-binding portion of the conjugate can bind.

82. A method for reducing the number of diseased cells in a system, comprising administering or delivering to the system a conjugate or composition as described in any one of claims 1 to 73, wherein the diseased cells express or contain an agent to which a target-binding portion of the conjugate can bind.

83. A method for delivering a payload to a target in a system, comprising applying or delivering to said system a conjugate or composition as described in any one of claims 1 to 73, wherein: The effective load portion of the conjugate is a portion of the effective load; and The target expresses or contains an agent that the target-binding portion of the conjugate can bind to.

84. The conjugate, composition, or kit as claimed in any one of claims 1 to 74, for use in treating a condition, disease, or ailment, or for use in a method as claimed in any one of the preceding claims.

85. The use of the conjugate, composition or kit as described in any one of claims 1 to 74 for the manufacture of a medicament for treating a condition, disease or ailment, or for the manufacture of a medicament for use in the method as described in any one of the preceding claims.

86. A method for preparing a conjugate or composition as described in any one of claims 1 to 73, comprising reacting a target binder and an effective loading agent, wherein: The target binder comprises a target binding portion and a target binder reactive group; The effective loading agent comprises an effective loading portion and an effective loading agent reactive group; and The target-binding reactive group reacts with the effective loading reactive group to optionally attach the effective loading portion to the target-binding portion via a connector.

87. The method of claim 84, wherein the effective loading agent is compound 1, compound 2, compound 3, compound 4, compound 5, compound 6, LP-A, LP-B, LP-C or LP-D.

88. A conjugate, composition, kit, method, or compound as described in any one of embodiments 1 to 813.

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