Nectin-4 antibody conjugates and uses thereof

Abstract

In one aspect, the present disclosure provides (a) an anti-Nectin-4 antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and the VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) a myeloid cell agonist; and (c) a myeloid cell agonist conjugate comprising the myeloid cell agonist and a linker covalently attached to the antibody.

Description

[Technical Field]

[0001] Sequence Listing Statement The Sequence Listing associated with this application is provided in text format in lieu of a paper copy and is hereby incorporated by reference. The name of the text file containing the Sequence Listing is 860234_402WO_SEQUENCE_LISTING.txt. The text file is 45.6 KB, was created on February 19, 2021, and has been submitted electronically via EFS-Web. [Background technology]

[0002] background Cancer is one of the leading causes of death in the United States. Traditional methods of cancer treatment, such as chemotherapy, surgery, or radiation therapy, tend to be highly toxic, nonspecific, or both, resulting in limited efficacy and harmful side effects. However, the immune system has the potential to be a powerful and specific tool in the fight against cancer. In many cases, tumors may specifically express genes whose products are necessary to induce or maintain a malignant state. These proteins may serve as antigen markers for developing more specific anticancer treatments, which can harness the power of both innate and adaptive immune responses. Activating these immune responses (e.g., myeloid cell activation) exclusively in the tumor microenvironment and lymphoid structures has the potential to be a powerful anticancer treatment that may be more effective than traditional methods of cancer treatment with fewer side effects. Summary of the Invention [Means for solving the problem]

[0003] A brief overview In one aspect, the present disclosure provides (a) an anti-Nectin-4 antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and the VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; (b) a myeloid cell agonist; and (c) a myeloid cell agonist conjugate comprising the myeloid cell agonist and a linker covalently attached to the antibody.

[0004] In certain embodiments, the conjugate has formula (I): [ka] (Wherein: A is an anti-Nectin-4 antibody or an antigen-binding fragment thereof, L is a linker; D x is a myeloid cell agonist, wherein the myeloid cell agonist is a TLR8 agonist; n is selected from 1 to 20; and z is selected from 1 to 20.

[0005] In a related aspect, the present disclosure provides a compound of the formula: [ka] (wherein the antibody is an anti-Nectin-4 antibody comprising light chain CDR1, CDR2, and CDR3 set forth in the amino acid sequence of the light chain variable region of SEQ ID NO: 14 or 13, and heavy chain CDR1, CDR2, and CDR3 set forth in the amino acid sequence of the heavy chain variable region of SEQ ID NO: 10, as determined by the Kabat index; L 3 -D is a linker-TLR8 agonist having the structure: [ka] (In the formula, RX* is a bond, succinimide moiety, or hydrolyzed succinimide moiety attached to a residue of the antibody construct, and R X * Above [ka] wherein R represents a point of attachment to a cysteine ​​residue of the antibody construct, or a salt thereof.

[0006] In another aspect, the present disclosure provides a pharmaceutical composition comprising a myeloid cell agonist conjugate disclosed herein and a pharmaceutically acceptable excipient.

[0007] In a further aspect, the present disclosure provides a method of treating cancer (e.g., a Nectin-4-expressing cancer), comprising administering to a subject in need thereof an effective amount of a myeloid cell agonist conjugate or pharmaceutical composition disclosed herein.

[0008] In certain embodiments, the myeloid cell agonist conjugate or pharmaceutical composition is administered subcutaneously.

[0009] In certain embodiments, the effective amount of the myeloid cell agonist conjugate is about 0.1 to about 100 mg / kg, preferably about 0.1 to about 25 mg / kg, and more preferably about 0.5 to about 20 mg / kg per treatment cycle or administration.

[0010] In certain embodiments, the myeloid cell agonist conjugate or pharmaceutical composition is administered subcutaneously, and the effective amount of the myeloid cell agonist conjugate is about 0.1 to about 100 mg / kg, preferably about 0.1 to about 25 mg / kg, and more preferably about 0.5 to about 20 mg / kg per treatment cycle or administration.

[0011] In another aspect, the present disclosure provides an isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to Nectin-4, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and the VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8.

[0012] In a further aspect, the present disclosure provides a conjugate comprising an anti-Nectin-4 antibody disclosed herein and a small molecule drug, such as a TLR8 agonist.

[0013] In another aspect, the present disclosure provides a pharmaceutical composition comprising an anti-Nectin-4 antibody disclosed herein or a conjugate comprising such an antibody and a small molecule drug, and a pharmaceutically acceptable carrier.

[0014] In another aspect, the present disclosure provides an isolated nucleic acid encoding an anti-Nectin-4 antibody, or the heavy chain, light chain, heavy chain variable region, or light chain variable region of an anti-Nectin-4 antibody. The present disclosure also provides a vector comprising the isolated nucleic acid, an isolated host cell comprising the isolated nucleic acid or vector, an isolated host cell that expresses the anti-Nectin-4 antibody disclosed herein, and a method for producing an anti-Nectin-4 antibody, comprising culturing the host cell disclosed herein under conditions suitable for expressing the antibody.

[0015] In a further aspect, the present disclosure provides a method for treating a Nectin-4-expressing cancer, the method comprising administering an effective amount of an anti-Nectin-4 antibody, a conjugate comprising an anti-Nectin-4 antibody and a small molecule drug, or a pharmaceutical composition comprising the antibody or conjugate, as disclosed herein, to a subject with a Nectin-4-expressing cancer.

[0016] In certain embodiments, the conjugate comprising an anti-Nectin-4 antibody and a small molecule drug, or the pharmaceutical composition thereof, is administered subcutaneously.

[0017] In certain embodiments, the effective amount of the conjugate is from about 0.1 to about 100 mg / kg, preferably from about 0.1 to about 25 mg / kg, and more preferably from about 0.5 to about 20 mg / kg per treatment cycle or per administration.

[0018] In certain embodiments, the conjugate or pharmaceutical composition is administered subcutaneously, and the effective amount of the conjugate is about 0.1 to about 100 mg / kg, preferably about 0.1 to about 25 mg / kg, and more preferably about 0.5 to about 20 mg / kg per treatment cycle or per administration. [Brief explanation of the drawings]

[0019] [Figure 1A] 1A-1C show that anti-nectin-4 antibodies and anti-nectin-4 antibody-TLR8 agonist immunoconjugates bind to (A) HEK-293 cells transfected with human nectin-4 (FIG. 1A); (B) HEK-293 cells transfected with cynomolgus monkey nectin-4 (FIG. 1B); and (C) MDA-MB-175-VII cells, a nectin-4-expressing tumor cell line (FIG. 1C). [Figure 1B] Same as above. [Figure 1C] Same as above.

[0020] [Figure 2A]Figures 2A-2B show TNF-α production in PBMC-nectin-4 tumor cell cocultures exposed to anti-nectin-4-TLR8 agonist conjugates but not unconjugated nectin-4-specific antibodies. PBMCs were cultured with (A) nectin-4-expressing MDA-MB-175-VII cells (Figure 2A) or (B) nectin-4-negative HEK-293 cells (Figure 2B) for 24 hours in the presence of equivalent titrated concentrations of anti-nectin-4-TLR8 agonists comparable to unconjugated anti-nectin-4 mAb or isotype control antibody conjugates. [Figure 2B] Same as above.

[0021] [Figure 3A] 3A-3B show that an anti-nectin-4-TLR7 agonist immunoconjugate (a surrogate for the human TLR8 agonist conjugate of the present disclosure, since only TLR7 (and not TLR8) is expressed in mouse myeloid cells) can induce TNF-α production in mouse bone marrow-derived macrophages (BMDMs) when co-cultured with HEK-293 human cells expressing nectin-4. [Figure 3B] Same as above.

[0022] [Figure 4] FIG. 4 is a Kaplan-Meier plot of mice bearing human Nectin-4-expressing EMT6 tumors treated with surrogates of Nectin-4 conjugates of the present disclosure.

[0023] [Figure 5A]Figures 5A and 5B show that in vivo treatment with a surrogate of the human Nectin4-TLR8 conjugate in tumor-bearing mice results in an increase in intratumoral chemokines (Figure 5A) and cytokines (Figure 5B). Mice bearing Nectin4-expressing EMT6 tumors were treated with a single dose of either D6C mIgG2a (unconjugated control) or D6C mIgG2a-Compound 4.1 conjugate, and tumors were harvested (A) on day 2 and the levels of the indicated chemokines in the tumors were assessed (Figure 5A) or (B) on day 5 and the levels of the indicated cytokines in the tumors were assessed (Figure 5B). Statistical significance was determined by the Mann-Whitney test. ***p<0.001, **p<0.01, *p<0.05. [Figure 5B] Same as above.

[0024] [Figure 6] FIG. 6 shows that the binding of TIGIT-Fc to Nectin-4-expressing tumor cells was blocked by the binding domain of hzD6.2C.

[0025] [Figure 7A] Figures 7A-7C represent tumor volume (in cubic millimeters) as a function of time (measured in days after treatment) after subcutaneous injection of mouse IgG2a isotype control (mIgG2a) (Figure 7A), anti-nectin-4 antibody alone (D6C mIgG2a) (Figure 7B), or anti-nectin-4-TLR7 agonist conjugate surrogate (D6C mIgG2a-Compound 4.1) (Figure 7C) into mice bearing human nectin-4-expressing EMT6 tumors. [Figure 7B] Same as above. [Figure 7C] Same as above.

[0026] [Figure 8] FIG. 8 depicts mean pre-dose and peak serum MCP-1 and IP-10 concentrations compared with dose levels in monkeys after subcutaneous administration of anti-Nectin-4-TLR8 agonist conjugate (D6.2C IgG1-Compound 2.14). DETAILED DESCRIPTION OF THE INVENTION

[0027] Detailed Description The present disclosure provides anti-Nectin-4 antibodies, myeloid cell agonist conjugates comprising anti-Nectin-4 antibodies, and pharmaceutical compositions comprising such antibodies and conjugates. The antibodies, conjugates, and pharmaceutical compositions disclosed herein are useful for treating cancer, either alone or in combination with other anti-cancer therapeutic agents.

[0028] The anti-nectin-4 antibodies provided herein and myeloid cell agonist conjugates comprising such antibodies can specifically bind to nectin-4-expressing cells. The myeloid cell agonist conjugates can also induce TNF-α production from human peripheral blood mononuclear cells (PBMCs) in the presence of nectin-4-expressing tumor cells, indicating that myeloid cells are activated by the conjugates of the present disclosure. Surprisingly, certain exemplary conjugates comprising a humanized anti-nectin-4 antibody were not only more potent in inducing TNF-α production from PBMCs compared to conjugates comprising a parent anti-nectin-4 antibody and compared to conjugates comprising a reference anti-nectin-4 antibody that cross-blocks the anti-nectin-4 antibody of the present disclosure, but also reached higher maximum TNF-α production levels.

[0029] The anti-Nectin-4 antibodies and myeloid cell agonist conjugates comprising such antibodies provided herein can also increase intratumoral levels of chemokines and cytokines, indicating that they can enhance the innate immune response driven by myeloid cell activation, which in turn can nucleate the adaptive immune response by indirectly activating T cells and NK cells via tumors.

[0030] In certain embodiments, the anti-Nectin-4 antibodies and myeloid cell agonist conjugates comprising such antibodies provided herein can block the binding of TIGIT to Nectin-4 expressed in tumor cells.

[0031] Before describing this disclosure in more detail, it may be helpful to its understanding to provide definitions of certain terms used herein. Additional definitions are set forth throughout this disclosure.

[0032] As used in this specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. The terms "a" and "an," as used herein, should be understood to refer to "one or more" of the listed members.

[0033] The use of the alternative (e.g., "or") should be understood to mean either one of the alternatives, both, or any combination thereof. As used herein, the terms "include" and "comprise" are used synonymously.

[0034] The phrase "at least one," when followed by a list of items or elements, refers to an open-ended set of one or more of the elements in the list, which may not necessarily include more than one element.

[0035] The term "about," when used herein in the context of a number, refers to a range that extends from less than 15% of that number to 15% greater than that number, with that number at the center. The term "about," when used in the context of a range, refers to an expanded range that extends from less than 15% of the lowest number listed in the range to 15% greater than the highest number listed in the range.

[0036] As used herein, any concentration range, percentage range, ratio range, or integer range should be understood to include any value (including integers or fractions) or subrange within the recited range, unless otherwise indicated.

[0037] As used herein, the term "antibody" refers to an immunoglobulin molecule that specifically binds to or is immunologically reactive with a specific antigen. Antibodies can include, for example, polyclonal antibodies, monoclonal antibodies, and genetically engineered antibodies, as well as antigen-binding fragments thereof. Antibodies can be, for example, murine, chimeric, humanized, heteroconjugate, bispecific, diabody, triabody, or tetrabody.

[0038] As used herein, "antigen-binding domain" or "antigen-binding fragment" refers to the region of a molecule that specifically binds to an antigen. The antigen-binding domain can be the antigen-binding portion of an antibody or antibody fragment. Antigen-binding fragments include, for example, Fab', F(ab')2, Fab, Fv, rIgG, scFv, hcAb (heavy chain antibody), single domain antibody, V HH , V NAR , sdAb, or nanobody.

[0039] As used herein, "Fc domain" refers to a domain derived from the Fc portion of an antibody, which is capable of specifically binding to an Fc receptor, such as an Fcγ receptor or an FcRn receptor.

[0040] As used herein, "identical" or "identity" refers to the similarity between a DNA, RNA, nucleotide, amino acid, or protein sequence and another DNA, RNA, nucleotide, amino acid, or protein sequence. Identity can be expressed in terms of the percentage of sequence identity of a first sequence to a second sequence. The percent (%) sequence identity with respect to a reference DNA sequence can be the percentage of DNA nucleotides in a candidate sequence that are identical to the DNA nucleotides in the reference DNA sequence after aligning the sequences. The percent (%) sequence identity with respect to a reference amino acid sequence can be the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the reference amino acid sequence after aligning the sequences and, if necessary, after introducing gaps to achieve the maximum percent sequence identity, not counting any conservative substitutions as part of the sequence identity. As used herein, percent sequence identity values ​​are generated using the NCBI BLAST 2.0 software as defined by Altschul et al., "Gapped BLAST and PSI-BLAST: a new generation of protein database search programs," Nucleic Acids Res. 2007, 25, 3389-3402, with parameters set to default values.

[0041] A "small molecule" is an organic compound having a molecular weight of less than 1500, or 100, or 900, or 750, or 600, or 500 daltons. A "small molecule drug" is a small molecule that has a therapeutic effect, such as treating a disease or disorder. In some embodiments, the small molecule drug is a small molecule agonist with an octanol-water partition coefficient (logP) in the range of 3-6, or 4-5, or 2-4. In some embodiments, the small molecule agonist has an octanol-water partition coefficient (logP) in the range of 200 Å or less. 2 Less than or equal to 150Å 2In some embodiments, the small molecule agonist has 5 or fewer, or 3 or fewer hydrogen bond donors and 10 or fewer, or 3 or fewer hydrogen bond acceptors. A small molecule is not a protein, polysaccharide, or nucleic acid. A "small molecule inhibitor" is a small molecule that inhibits the activity of another molecule, such as a protein (e.g., PD-L1). Small molecule inhibitors include small molecule antagonists (i.e., small molecules that reduce the effect of an agonist).

[0042] As used herein, "specifically binds" and the like refers to a specific association or binding between an antigen-binding domain and an antigen, as compared to the interaction of the antigen-binding domain with a different antigen (i.e., non-specific binding). In some embodiments, an antigen-binding domain that recognizes or specifically binds to an antigen has a specific binding affinity of <100 nM, <10 nM, <1 nM, <0.1 nM, <0.01 nM, or <0.001 nM (e.g., 10 -8 M or less, e.g., 10 -8 M~10 -13 M, e.g., 10 -9 M~10 -13 Specific binding does not require that the antigen-binding domain does not associate or bind to any other antigen, but rather that it preferentially associates or binds to the antigen compared to association or binding to unrelated antigens.

[0043] As used herein, "nectin-4," also known as poliovirus receptor-related protein 4 (PVRL4), LNIR, PRR4, and EDSS1, is a Ca 2+Nectin-4 is a member of the nectin subfamily of immunoglobulin-like adhesion molecules involved in cell-cell adhesion. Nectin binds to the actin cytoskeleton via the adaptor protein afadin (AFDN) and is an important component of adherens junctions. Nectin-4 contains two immunoglobulin-like (Ig-like) C2-type domains and one Ig-like V-type domain. It can be a single-pass type I membrane protein, or its soluble form is generated by proteolytic cleavage at the cell surface by the metalloproteinase ADAM17 / TACE. Nectin-4 is overexpressed in multiple human cancers, including, but not limited to, triple-negative breast cancer, bladder cancer, urothelial cancer, ovarian cancer, cervical cancer, endometrial cancer, lung cancer, skin cancer, and esophageal cancer, and its aberrant expression may be associated with cancer progression and poor prognosis. Examples of nectin-4 include mammalian nectin-4 proteins from mice, rats, rabbits, guinea pigs, pigs, sheep, dogs, non-human primates, and humans. In some embodiments, nectin-4 is human nectin-4 (encoded by GenBank accession number AF426163) or mouse nectin-4 (see Reymond et al., Journal of Biological Chemistry 276:43205-15, 2001).

[0044] As used herein, "immune cells" refers to T cells, B cells, NK cells, NKT cells, or antigen-presenting cells. In some embodiments, the immune cells are T cells, B cells, NK cells, or NKT cells. In some embodiments, the immune cells are antigen-presenting cells. In some embodiments, the immune cells are not antigen-presenting cells.

[0045] As used herein, an "immunostimulatory compound" is a compound that activates or stimulates immune cells, such as myeloid cells or APCs.

[0046] As used herein, "myeloid cells" refers to dendritic cells, macrophages, monocytes, and myeloid-derived suppressor cells (MDSCs).

[0047] As used herein, "myeloid cell agonist" refers to a compound that activates or stimulates an immune response by myeloid cells.

[0048] As used herein, "benzazepine compound" refers to a small molecule compound containing a benzazepine moiety, where the benzazepine moiety is a benzene ring fused to a seven-membered ring containing one or two nitrogen ring members. In addition to the bond where the ring is fused to the benzene ring, the seven-membered ring contains two double bonds (e.g., an azepine or diazepine ring), one double bond (e.g., a dihydroazepine or dihydro-diazepine ring), or no double bonds (e.g., a tetrahydroazepine, azepane, tetrahydrodiazepine, or diazepane ring). The benzazepine moiety is optionally substituted. In some embodiments, the benzazepine moiety is an optionally substituted 4,5-dihydro-3H-benzo[b]azepine. In some embodiments, the benzazepine moiety has the structure: [ka] (In the formula, [ka] is a double bond or a single bond; L 2 -X 2 -, -X 2 -C 1~6 Alkylene-X 2 -, -X 2 -C 2~6 Alkenylene-X 2 - and -X 2 -C 2~6 Alkynylene-X 2 -, each of which in alkylene, alkenylene, or alkynylene is selected from one or more R 12 with substitution as necessary; X 2 In each occurrence, -O-, -S-, -N(R 10)-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 ), -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O), -OS(O)2-, -S(O)2O, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, and -N(R 10 )S(O)N(R 10 )-independently selected from; R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 independently selected from carbocycle and 3- to 10-membered heterocycle, wherein R 12 Each C in 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings are not substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 4 -OR 10 , -N(R 10 )2, -C(O)N(R10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 selected from carbocycles and 3- to 12-membered heterocycles, where R 4 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 10represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, -C 1~10 Haloalkyl, -OC 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; The moiety is optionally substituted at any position. It has.

[0049] The term "salt" or "pharmaceutically acceptable salt" refers to salts derived from various organic and inorganic counterions well known in the art. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, trifluoroacetic acid, etc. Pharmaceutically acceptable base addition salts can be formed with inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Organic bases from which salts can be derived include, for example, naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like, specifically substituted amines of primary, secondary, and tertiary amines, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, the pharmaceutically acceptable base addition salt is selected from ammonium, potassium, sodium, calcium, and magnesium salts.

[0050] "C x~y The term "alkyl," when used in connection with a chemical moiety such as alkyl, alkenyl, or alkynyl, is meant to include groups containing x to y carbons in the chain. For example, "C 1~6 The term "alkyl" refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain and branched-chain alkyl groups, containing 1 to 6 carbons. x~y The term alkylene- refers to a substituted or unsubstituted alkylene chain having x to y carbon atoms in the alkylene chain. For example, -C 1~6 Alkylene- may be selected from methylene, ethylene, propylene, butylene, pentylene, and hexylene, any one of which is optionally substituted.

[0051] "C x~y alkenyl" and "C x~y The term "alkynyl" refers to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double or triple bond respectively. x~y The term alkenylene- refers to a substituted or unsubstituted alkenylene chain having x to y carbon atoms in the chain. For example, -C 2~6 Alkenylene- may be selected from ethenylene, propenylene, butenylene, pentenylene, and hexenylene, any one of which is optionally substituted. The alkenylene chain may have one double bond or two or more double bonds in the alkenylene chain. -C x~y The term alkynylene- refers to a substituted or unsubstituted alkynylene chain having x to y carbon atoms in the alkynylene chain. For example, -C 2~6 Alkynylene- may be selected from ethynylene, propynylene, butynylene, pentynylene, and hexynylene, any one of which is optionally substituted. The alkynylene chain may have one triple bond or two or more triple bonds within the alkynylene chain.

[0052] "Alkylene" refers to a divalent hydrocarbon chain, such as methylene, ethylene, propylene, butylene, etc., that connects the rest of the molecule to a radical group. The divalent hydrocarbon chain, consisting solely of carbon and hydrogen, contains no unsaturation, and preferably has 1 to 12 carbon atoms. The alkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group are through the terminal carbons, respectively. In other embodiments, alkylene contains 1 to 5 carbon atoms (i.e., C1-C5 alkylene). In other embodiments, alkylene contains 1 to 4 carbon atoms (i.e., C1-C4 alkylene). In other embodiments, alkylene contains 1 to 3 carbon atoms (i.e., C1-C3 alkylene). In other embodiments, alkylene contains 1 to 2 carbon atoms (i.e., C1-C2 alkylene). In other embodiments, alkylene contains 1 carbon atom (i.e., C1 alkylene). In other embodiments, an alkylene contains 5 to 8 carbon atoms (i.e., a C5-C8 alkylene). In other embodiments, an alkylene contains 2 to 5 carbon atoms (i.e., a C2-C5 alkylene). In other embodiments, an alkylene contains 3 to 5 carbon atoms (i.e., a C3-C5 alkylene). Unless stated otherwise specifically in the specification, an alkylene chain is optionally substituted with one or more substituents, such as those substituents described herein.

[0053] "Alkenylene" refers to a divalent hydrocarbon chain, consisting solely of carbon and hydrogen, containing at least one carbon-carbon double bond, preferably having 2 to 12 carbon atoms, connecting the rest of the molecule to a radical group. The alkenylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkenylene chain to the rest of the molecule and to the radical group are through the terminal carbons, respectively. In other embodiments, the alkenylene contains 2 to 5 carbon atoms (i.e., a C2-C5 alkenylene). In other embodiments, the alkenylene contains 2 to 4 carbon atoms (i.e., a C2-C4 alkenylene). In other embodiments, the alkenylene contains 2 to 3 carbon atoms (i.e., a C2-C3 alkenylene). In other embodiments, the alkenylene contains 2 carbon atoms (i.e., a C2 alkenylene). In other embodiments, an alkenylene contains 5 to 8 carbon atoms (i.e., a C5-C8 alkenylene). In other embodiments, an alkenylene contains 3 to 5 carbon atoms (i.e., a C3-C5 alkenylene). Unless stated otherwise specifically in the specification, an alkenylene chain is optionally substituted with one or more substituents, such as those substituents described herein.

[0054] "Alkynylene" refers to a divalent hydrocarbon chain connecting the rest of the molecule to a radical group, consisting solely of carbon and hydrogen, containing at least one carbon-carbon triple bond, and preferably having 2 to 12 carbon atoms. The alkynylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the alkynylene chain to the rest of the molecule and to the radical group are through the terminal carbons, respectively. In other embodiments, the alkynylene contains 2 to 5 carbon atoms (i.e., a C2-C5 alkynylene). In other embodiments, the alkynylene contains 2 to 4 carbon atoms (i.e., a C2-C4 alkynylene). In other embodiments, the alkynylene contains 2 to 3 carbon atoms (i.e., a C2-C3 alkynylene). In other embodiments, the alkynylene contains 2 carbon atoms (i.e., a C2 alkynylene). In other embodiments, an alkynylene contains 5 to 8 carbon atoms (i.e., a C5-C8 alkynylene). In other embodiments, an alkynylene contains 3 to 5 carbon atoms (i.e., a C3-C5 alkynylene). Unless stated otherwise specifically in the specification, an alkynylene chain is optionally substituted with one or more substituents, such as those substituents described herein.

[0055] "Heteroalkylene" refers to a divalent hydrocarbon chain containing at least one heteroatom in the chain, containing no unsaturation, and preferably having 1 to 12 carbon atoms and 1 to 6 heteroatoms, e.g., -O-, -NH-, -S-. The heteroalkylene chain is attached to the rest of the molecule through a single bond and to the radical group through a single bond. The points of attachment of the heteroalkylene chain to the rest of the molecule and to the radical group are through the terminal atoms of the chain. In other embodiments, heteroalkylene contains 1 to 5 carbon atoms and 1 to 3 heteroatoms. In other embodiments, heteroalkylene contains 1 to 4 carbon atoms and 1 to 3 heteroatoms. In other embodiments, heteroalkylene contains 1 to 3 carbon atoms and 1 to 2 heteroatoms. In other embodiments, heteroalkylene contains 1 to 2 carbon atoms and 1 to 2 heteroatoms. In other embodiments, heteroalkylene contains 1 carbon atom and 1 to 2 heteroatoms. In another embodiment, a heteroalkylene contains 5 to 8 carbon atoms and 1 to 4 heteroatoms. In another embodiment, a heteroalkylene contains 2 to 5 carbon atoms and 1 to 3 heteroatoms. In another embodiment, a heteroalkylene contains 3 to 5 carbon atoms and 1 to 3 heteroatoms. Unless stated otherwise specifically in the specification, a heteroalkylene chain is optionally substituted with one or more substituents, such as those substituents described herein.

[0056] The term "carbocycle" as used herein refers to a saturated, unsaturated, or aromatic ring in which each atom of the ring is carbon. Carbocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Each ring of a bicyclic carbocycle can be selected from saturated, unsaturated, and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, can be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Bicyclic carbocycles include any combination of saturated, unsaturated, and aromatic bicyclic rings, where valence allows. Bicyclic carbocycles include any combination of ring sizes, such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. Exemplary carbocyclic rings include cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, phenyl, indanyl, and naphthyl. The term "unsaturated carbocyclic ring" refers to a carbocyclic ring having at least one degree of unsaturation and excluding aromatic carbocyclic rings. Examples of unsaturated carbocyclic rings include cyclohexadiene, cyclohexene, and cyclopentene.

[0057] The term "heterocycle," as used herein, refers to a saturated, unsaturated, or aromatic ring containing one or more heteroatoms. Exemplary heteroatoms include N, O, Si, P, B, and S atoms. Heterocycles include 3- to 10-membered monocyclic rings, 6- to 12-membered bicyclic rings, and 6- to 12-membered bridged rings. Bicyclic heterocycles include any combination of saturated, unsaturated, and aromatic bicyclic rings, where valence allows. In an exemplary embodiment, an aromatic ring, such as pyridyl, may be fused to a saturated or unsaturated ring, such as cyclohexane, cyclopentane, morpholine, piperidine, or cyclohexene. Bicyclic heterocycles include any combination of ring sizes, such as 4-5 fused ring systems, 5-5 fused ring systems, 5-6 fused ring systems, 6-6 fused ring systems, 5-7 fused ring systems, 6-7 fused ring systems, 5-8 fused ring systems, and 6-8 fused ring systems. The term "unsaturated heterocycle" refers to a heterocycle having at least one degree of unsaturation and excluding aromatic heterocycles. Examples of unsaturated heterocycles include dihydropyrrole, dihydrofuran, oxazoline, pyrazoline, and dihydropyridine.

[0058] The term "heteroaryl" includes aromatic monocyclic ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, where the ring structure contains at least one heteroatom, preferably 1-4 heteroatoms, more preferably 1 or 2 heteroatoms. The term "heteroaryl" also includes polycyclic ring systems having two or more rings in which two or more carbons are common to two adjacent rings, where at least one of the rings is heteroaromatic; for example, the other rings can be aromatic or non-aromatic, carbocyclic, or heterocyclic. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine.

[0059] The term "substituted" refers to moieties having substituents replacing a hydrogen on one or more carbons or substitutable heteroatoms, e.g., -NH-, of a structure. It will be understood that "substituted" or "substituted with" includes the implicit proviso that such substitution is subject to the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, i.e., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, and the like. In certain embodiments, substituted refers to moieties having substituents replacing two hydrogen atoms on the same carbon atom, such as replacing two hydrogen atoms on a single carbon with an oxo, imino, or thioxo group. As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and nonaromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this disclosure, the heteroatoms such as nitrogen may have hydrogen substituents and / or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms.

[0060] In some embodiments, the substituents include any of the substituents described herein, for example, halogen, hydroxy, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO), imino (=NH), oximo (=N-OH), hydrazino (=N-NH), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b-C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2), and -R b -S(O) t N(R a )2 (wherein t is 1 or 2); and alkyl, alkenyl, alkynyl, aryl, aralkyl, aralkenyl, aralkynyl, cycloalkyl, cycloalkylalkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, and heteroarylalkyl (any of which may be alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO2), imino (=NH), oximo (=N-OH), hydrazine (=N-NH2), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(R a)2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2), and -R b -S(O) t N(R a ) 2 (wherein t is 1 or 2) (wherein each R a is independently selected from hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl; each R a is, if valence allows, alkyl, alkenyl, alkynyl, halogen, haloalkyl, haloalkenyl, haloalkynyl, oxo (=O), thioxo (=S), cyano (-CN), nitro (-NO2), imino (=NH), oximo (=N-OH), hydrazine (=N-NH2), -R b -OR a , -R b -OC(O)-R a , -R b -OC(O)-OR a , -R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a , -R b -C(O)OR a , -R b -C(O)N(Ra )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a , -R b -N(R a )C(O)R a , -R b -N(R a )S(O) t R a (wherein t is 1 or 2), -R b -S(O) t R a (wherein t is 1 or 2), -R b -S(O) t OR a (wherein t is 1 or 2), and -R b -S(O) t N(R a )2 (wherein t is 1 or 2); each R b are independently selected from a direct bond, or a linear or branched alkylene, alkenylene, or alkynylene chain, and each R c may include a group optionally substituted by a group which is a linear or branched alkylene, alkenylene or alkynylene chain.

[0061] It will be understood by those skilled in the art that, where appropriate, the substituents themselves can be substituted. Unless specifically designated "unsubstituted," reference to a chemical moiety herein is understood to include substituted variants. For example, reference to a "heteroaryl" group or moiety implicitly includes both substituted and unsubstituted variants.

[0062] In addition, it should be understood that individual compounds (e.g., proteins) or groups of compounds (e.g., domains, regions, or peptide components) derived from various combinations of the structures and substituents described herein are disclosed in this application to the same extent as if each compound or group of compounds were individually described. As such, selection of a particular structure or particular substituents is within the scope of this disclosure.

[0063] Chemical substances having carbon-carbon or carbon-nitrogen double bonds may exist in Z or E forms (or cis or trans forms). Furthermore, some chemical substances may exist in various tautomeric forms. Unless otherwise specified, chemical substances described herein are intended to include all Z, E, and tautomeric forms as well.

[0064] "Tautomer" refers to a molecule in which a proton can shift from one atom of the molecule to another atom of the same molecule. The compounds presented herein exist as tautomers in certain embodiments. In situations where tautomerization is possible, a chemical equilibrium of tautomers will exist. The exact ratio of tautomers depends on several factors, including physical conditions, temperature, solvent, and pH. Some examples of tautomeric equilibrium include: [ka] Examples include:

[0065] As used herein, "conjugate" refers to an antibody or antigen-binding fragment thereof attached, optionally via a linker, to at least one immunostimulatory compound.

[0066] The phrases "intravenous administration" and "intravenously administered," as used herein, refer to the injection or infusion of a conjugate into a subject's vein.

[0067] The phrases "slow intravenous infusion" and "slow IV infusion," as used herein, refer to an intravenous infusion that results in a Tmax of about 4 hours or longer.

[0068] The phrases "subcutaneous administration," "administering subcutaneously," and the like refer to administration of the conjugate into the subcutaneous tissue of a subject. For clarity, subcutaneous administration is distinguished from intratumoral injection into a tumor or cancerous lesion located subcutaneously.

[0069] The phrase "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. Anti-nectin-4 antibody

[0070] In one aspect, an antibody (e.g., an isolated monoclonal antibody), or an antigen-binding fragment thereof, that specifically binds to Nectin-4, also referred to as an anti-Nectin-4 antibody, is provided.

[0071] In various embodiments, an antibody or antigen-binding fragment thereof comprises two light chain polypeptides (light chains) and two heavy chain polypeptides (heavy chains) covalently held together by disulfide bonds.

[0072] A heavy chain typically comprises a heavy chain variable region (VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2, and CH3. Non-limiting exemplary heavy chain constant regions include the constant regions of human IgG1, human IgG2, human IgG3, and human IgG4. In some embodiments, the antibodies provided herein comprise an IgG1 constant region.

[0073] A light chain typically comprises a light chain variable region (VL) and a light chain constant region. Non-limiting exemplary light chain constant regions include kappa and lambda constant regions. A non-limiting exemplary human kappa constant region is set forth in SEQ ID NO: 20. Another exemplary light chain constant region is the mouse kappa constant region set forth in SEQ ID NO: 22.

[0074] The constant domains provide the general framework of an antibody and are not directly involved in binding the antibody to an antigen, but may be involved in various effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC), ADCP (antibody-dependent cellular phagocytosis), CDC (complement-dependent cytotoxicity), and complement fixation, binding to Fc receptors (e.g., CD16, CD32, FcRn), longer in vivo half-life compared to polypeptides lacking the Fc region, protein A binding, and possibly even placental passage (see Capon et al., Nature 337:525, 1989). As used herein, "Fc region constant domain portion" or "Fc region portion" refers to the heavy chain constant region segment of an Fc fragment ("fragment crystallizable" region or Fc region) derived from an antibody, which may include one or more constant domains, such as CH2, CH3, CH4, or any combination thereof. In certain embodiments, the Fc region portion includes the CH2 and CH3 domains of an IgG, IgA, or IgD antibody, and any combination thereof, or the CH3 and CH4 domains of an IgM or IgE antibody, and any combination thereof.

[0075] The Fc region or domain can interact with different types of FcR. Different types of FcR can include, for example, FcγRI, FcγRIIA, FcγRIIB, FcγRIIIA, FcγRIIIB, FcαRI, FcμR, FcεRI, FcεRII, and FcRn. FcR can be located on the membrane of certain immune cells, including, for example, B lymphocytes, natural killer cells, macrophages, neutrophils, follicular dendritic cells, eosinophils, basophils, platelets, and mast cells. When FcR binds to the Fc domain, it can initiate functions including, for example, clearance of antigen-antibody complexes via receptor-mediated endocytosis, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), trogocytosis, trogoptosis, and ligand-induced transduction of signals across the cell membrane, which can lead to secretion, exocytosis, and alteration of cellular metabolism. FcRs can deliver signals when they aggregate with antibodies and multivalent antigens on the cell surface. Aggregation of FcRs with immunoreceptor tyrosine-based activation motifs (ITAMs) can, in turn, activate the SRC and SYK family of tyrosine kinases. ITAMs contain a double-repeated YxxL sequence flanked by seven variable residues. SRC and SYK kinases can connect the transduced signals to a common activation pathway.

[0076] In some embodiments, an Fc region or domain may exhibit reduced binding affinity to one or more Fc receptors. In some embodiments, an Fc region or domain may exhibit reduced binding affinity to one or more Fcγ receptors. In some embodiments, an Fc region or domain may exhibit reduced binding affinity to the FcRn receptor. In some embodiments, an Fc region or domain may exhibit reduced binding affinity to the Fcγ and FcRn receptors. In some embodiments, an Fc domain is an Fc null domain or region. As used herein, "Fc null" refers to a domain that exhibits weak to no binding to any of the Fc gamma receptors. In some embodiments, an Fc null domain or region exhibits at least about a 1000-fold reduced binding affinity (e.g., increased Kd) to an Fcγ receptor.

[0077] The Fc region or domain may have one or more, two or more, three or more, four or more, or up to five amino acid substitutions that reduce binding of the Fc region or domain to an Fc receptor. In some embodiments, the Fc region or domain exhibits reduced binding to FcγRI (CD64), FcγRIIA (CD32), FcγRIIIA (CD16a), FcγRIIIB (CD16b), or any combination thereof. To reduce the binding affinity of the Fc region or domain to an Fc receptor, the Fc region or domain may contain one or more amino acid substitutions that have the effect of reducing the affinity of the Fc domain or region for an Fc receptor. In certain embodiments, the Fc region or domain is an IgG1, and the one or more substitutions in the Fc region or domain comprise any one or more of the IgG1 heavy chain mutations corresponding to E233P, L234V, L234A, L235A, L235E, ΔG236, G237A, E318A, K320A, K322A, A327G, A330S, or P331S according to the EU index of Kabat numbering.

[0078] In some embodiments, the Fc region or domain may comprise the sequence of an IgG1 isoform that has been modified from the wild-type IgG1 sequence. The modifications may include substitutions at two or more amino acid residues, such as five different amino acid residues comprising L235V / F243L / R292P / Y300L / P396L (IgG1VLPLL) according to the EU index of Kabat numbering. The modifications may include substitutions at two or more different amino acid residues, such as two different amino acid residues comprising S239D / I332E (IgG1DE) according to the EU index of Kabat numbering. The modifications may include substitutions at two or more different amino acid residues, such as three different amino acid residues comprising S298A / E333A / K334A (IgG1AAA) according to the EU index of Kabat numbering. Non-limiting exemplary IgG1 constant regions are set forth in SEQ ID NOs: 18 and 19. In certain other embodiments, the antibodies provided herein comprise a murine IgG2a heavy chain constant region set forth in SEQ ID NO: 21.

[0079] The antibody or Fc domain may be modified to acquire or improve at least one constant region-mediated biological effector function compared to the unmodified antibody or Fc domain, for example, to enhance FcγR interaction. In certain embodiments, the modification can increase CD32b binding (and support trans-delivery in PBMC assays) and includes substitutions at S267L and E329F (IgG1LF, also known as the SELF double mutation) according to the EU index of Kabat numbering. For example, antibodies having constant regions that bind to FcγRIIA, FcγRIIB, and / or FcγRIIIA with higher affinity than the corresponding wild-type constant regions can be produced according to the methods described herein. Fc domains that bind to FcγRIIA, FcγRIIB, and / or FcγRIIIA with higher affinity than the corresponding wild-type Fc domains can be produced according to the methods described herein.

[0080] In certain embodiments, the Fc region or domain found in the anti-Nectin-4 antibodies of the present disclosure will be capable of mediating one or more of these effector functions, or will lack one or more or all of these activities, or will have one or more of the effector activities increased, e.g., by means of one or more mutations, compared to an unmodified Fc region or domain.

[0081] The antigen recognition region of an antibody variable domain typically contains six complementarity determining regions (CDRs) or hypervariable regions, which are within the framework of the N-terminal heavy and light chain variable regions of two heavy and two light chains.

[0082] In some embodiments, the antigen-binding domain comprises light chain complementarity determining region 1 (LCDR1), light chain complementarity determining region 2 (LCDR2), light chain complementarity determining region 3 (LCDR3), heavy chain complementarity determining region 1 (HCDR1), heavy chain complementarity determining region 2 (HCDR2), and heavy chain complementarity determining region 3 (HCDR3). In some embodiments, the antibody may be a heavy chain-only antibody, in which case the antigen-binding domain comprises HCDR1, HCDR2, and HCDR3, and the antibody lacks a light chain.

[0083] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 9, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 11, provided that the amino acid sequences of the VH-CDRs (i.e., SEQ ID NOs: 1-3) and VL-CDRs (i.e., SEQ ID NOs: 4, 7, and 8) are unchanged. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from SEQ ID NO: 23, and a light chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 25, provided that the amino acid sequences of the VH-CDR and VL-CDR are unchanged.

[0084] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising an amino acid sequence selected from SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising an amino acid sequence comprising SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 10, and a light chain variable region (VL) comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from SEQ ID NOs: 12-17, provided that the amino acid sequences of the VH-CDR and VL-CDR are unchanged. In some embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 24, and a light chain comprising an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to an amino acid sequence selected from SEQ ID NOs: 26-31, provided that the amino acid sequences of the VH-CDR and VL-CDR are unchanged.

[0085] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 12, or both (a) and (b).

[0086] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 4, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In certain embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 13, or both (a) and (b).

[0087] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 5, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 14, or both (a) and (b).

[0088] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 15, or both (a) and (b).

[0089] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 16, or both (a) and (b).

[0090] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises a heavy chain CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3, a light chain CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 5, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8. In some such embodiments, the antibody or antigen-binding fragment thereof comprises (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10, (b) a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 17, or both (a) and (b).

[0091] In some embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof comprises: (a) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 9 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 11; or (b) a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 10 and a light chain variable region (VL) comprising an amino acid sequence selected from SEQ ID NOs: 12 to 17.

[0092] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:12.

[0093] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:13.

[0094] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:14.

[0095] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:15.

[0096] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:16.

[0097] In some such embodiments, the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:10 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:17.

[0098] In some embodiments, the anti-Nectin-4 antibody comprises (a) a heavy chain comprising the amino acid sequence of SEQ ID NO: 23 and a light chain comprising the amino acid sequence of SEQ ID NO: 25; or (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 24 and a light chain comprising an amino acid sequence selected from SEQ ID NOs: 26 to 31.

[0099] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:26.

[0100] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:27.

[0101] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:28.

[0102] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:29.

[0103] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:30.

[0104] In some such embodiments, the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:31.

[0105] In any of the foregoing embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof is conjugated with a small molecule drug to form an antibody-drug conjugate. In certain embodiments, the small molecule drug is a myeloid cell agonist (e.g., a TLR8 agonist) disclosed herein, thus forming a myeloid cell agonist conjugate.

[0106] Anti-Nectin-4 antibodies or antigen-binding fragments thereof can be chimeric or humanized. Chimeric and humanized forms of non-human (e.g., murine) antibodies can be intact (full-length) chimeric immunoglobulins, immunoglobulin chains, or antigen-binding fragments thereof (such as Fv, Fab, Fab', F(ab')2, or other target-binding subdomains of antibodies), which can contain sequences derived from non-human immunoglobulins. Generally, a humanized antibody or antigen-binding fragment thereof can comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework (FR) regions are those of a human immunoglobulin sequence. A humanized antibody can also comprise at least a portion of an immunoglobulin constant region (Fc), an Fc domain, typically of a human immunoglobulin sequence.

[0107] The anti-Nectin-4 antibodies or antigen-binding fragments thereof described herein may be human antibodies. As used herein, "human antibodies" can include, for example, antibodies having the amino acid sequence of a human immunoglobulin and can include antibodies isolated from a human immunoglobulin library or from animals transgenic for one or more human immunoglobulins that typically do not express endogenous immunoglobulins. Human antibodies can also be produced using transgenic mice that are unable to express functional endogenous immunoglobulins but can express human immunoglobulin genes. Fully human antibodies that recognize a selected epitope can be generated using guided selection. In this approach, a selected non-human monoclonal antibody, e.g., a mouse antibody, is used to guide the selection of fully human antibodies that recognize the same epitope.

[0108] The anti-Nectin-4 antibodies or antigen-binding fragments thereof described herein may be bispecific antibodies or dual variable domain antibodies (DVDs). Bispecific and DVD antibodies are monoclonal, often human or humanized, antibodies that have binding specificities for at least two different antigens, one of which is Nectin-4.

[0109] The anti-Nectin-4 antibodies or antigen-binding fragments thereof described herein can be derivatized or otherwise modified. For example, derivatized antibodies can be modified by glycosylation, acetylation, pegylation, phosphorylation, amidation, derivatization with known protecting / blocking groups, proteolytic cleavage, etc. Antibody CDR

[0110] The CDR sequences of Nectin-4 antibodies can be determined by one or more methods, including Kabat, Chothia, AbM, Contact, IMGT, and AHo (see Table A below). Unless otherwise specified herein, CDR sequences are determined according to the Kabat method. Reference to the numbering of variable regions or CDRs in Kabat refers to the numbering of amino acid positions in Kabat, or CDR sequences determined according to the Kabat method, and variations thereof, referring to the numbering system used for the heavy or light chain variable regions of the antibody compilation in Kabat et al. ((1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortening or insertion into the FRs or CDRs of the variable domain. For example, a heavy chain variable domain may include a single amino acid insertion after residue 52 (residue 52a according to Kabat) and three inserted residues after residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues may be determined for a given antibody by alignment of regions of sequence homology with an antibody having a "standard" Kabat numbered sequence. The Kabat numbering system is generally used when referring to residues in the variable domain (approximately residues 1-107 of the light chain and residues 1-113 of the heavy chain) (e.g., Kabat et al., supra).

[0111] The "EU numbering system" or "EU index" is commonly used when referring to residues in immunoglobulin heavy chain constant regions (e.g., the EU index reported in Kabat et al., supra). "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody.

[0112] Other numbering systems are described by, for example, AbM (Oxford Molecular's AbM antibody modeling software (see, e.g., Antibody Engineering Vol. 2 (Kontermann and Dithel eds., 2d ed. 2010)), Chothia (see, Chothia and Lesk, 1987, J. Mol. Biol. 196:901-17), Contact, IMGT (see, ImMunoGeneTics (IMGT) Information System® (Lafranc et al., 2003, Dev. Comp. Immunol. 27(1):55-77)), and AHon (see, Honegger and Pluckthun, 2001, J. Mol. Biol. 309: 657-70)) and are well understood by those of skill in the art.

[0113] In certain embodiments, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a heavy chain variable region (VH) comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and (b) a light chain variable region (VL) comprising a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8.

[0114] In a further embodiment, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a VH comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 33, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 35; and (b) a VL comprising a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 36, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 37, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 38.

[0115] In still further embodiments, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a VH comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 39, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 41; and (b) a VL comprising a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 42, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 44.

[0116] In still further embodiments, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a VH comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 45, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 46, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; and (b) a VL comprising a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 48, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 49, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 50.

[0117] In still further embodiments, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a VH comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and (b) a VL comprising a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 54, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 55, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 56.

[0118] In still further embodiments, the anti-Nectin-4 antibody of the present disclosure is composed of (a) a VH comprising a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 57, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 58, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 59; and (b) a VL comprising a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 60, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 62. [Table A] Nucleic acids, vectors, and host cells

[0119] The present disclosure provides isolated nucleic acids encoding the anti-Nectin-4 antibodies or antigen-binding fragments thereof described herein. In some embodiments, the nucleic acids encoding the anti-Nectin-4 antibodies or antigen-binding fragments thereof are codon-optimized to enhance or maximize expression in certain types of cells (e.g., Scholten et al., Clin. Immunol. 119: 135-145, 2006). As used herein, a "codon-optimized" polynucleotide is a heterologous polypeptide having codons modified with silent mutations that correspond to the abundance of tRNA levels in the host cell.

[0120] In some embodiments, the nucleic acid molecule comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein, wherein two or more chains or regions are separated by a cleavage site (e.g., antibody heavy and light chains, or V H and V LIn some embodiments, the cleavage site encodes a self-cleaving amino acid sequence comprising the 2A peptide from porcine teschovirus-1 (P2A), equine rhinitis A virus (E2A), tosea asiavirus (T2A), foot-and-mouth disease virus (F2A), or any combination thereof (see, e.g., Kim et al., PLOS One 6:e18556, 2011, the 2A nucleic acid and amino acid sequences of which are incorporated herein by reference in their entirety).

[0121] In another aspect, an expression construct is provided comprising a nucleic acid encoding an anti-Nectin-4 antibody or antigen-binding fragment thereof described herein. In some embodiments, the nucleic acid may be operably linked to an expression control sequence. As used herein, "expression construct" refers to a DNA construct containing a nucleic acid molecule operably linked to a suitable control sequence capable of effecting expression of the nucleic acid molecule in a suitable host. The expression construct may be present in a vector (e.g., a bacterial vector, a viral vector) or may be integrated into a genome. The term "operably linked" refers to the association of two or more nucleic acids on a single polynucleotide fragment such that one function is affected by the other. For example, a promoter is operably linked to a coding sequence if it is capable of affecting the expression of that coding sequence (i.e., the coding sequence is under the transcriptional control of the promoter). The term "expression control sequence" (also called a regulatory sequence) refers to a nucleic acid sequence that effects the expression and processing of coding sequences to which it is operably linked. For example, expression control sequences may include transcription initiation, termination, promoter, and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (i.e., Kozak consensus sequences); sequences that enhance protein stability; and sequences that may enhance protein secretion.

[0122] In some embodiments, the nucleic acid or expression construct encoding the anti-Nectin-4 antibody or antigen-binding fragment thereof is present in a vector. A "vector" is a nucleic acid molecule capable of transporting another nucleic acid. Vectors can be, for example, plasmids, cosmids, viruses, RNA vectors, or linear or circular DNA or RNA molecules, and they may contain chromosomal, non-chromosomal, semisynthetic, or synthetic nucleic acids. Exemplary vectors are capable of autonomous replication (episomal vectors) or expression (expression vectors) of the nucleic acid to which they are linked. Exemplary viral vectors include retrovirus, adenovirus, parvovirus (for example, adeno-associated virus), coronavirus, negative-strand RNA virus, such as orthomyxovirus (for example, influenza virus), rhabdovirus (for example, rabies and vesicular stomatitis virus), paramyxovirus (for example, measles and Sendai virus), positive-strand RNA virus, such as picornavirus and alphavirus, and double-stranded DNA virus, such as adenovirus, herpesvirus (for example, herpes simplex virus type 1 and type 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (for example, vaccinia, fowlpox and canarypox).Other viruses include, for example, Norwalk virus, togavirus, flavivirus, reovirus, papovavirus, hepadnavirus, and hepatitis virus. Examples of retroviruses include avian leukosis sarcoma viruses, mammalian type C, type B, type D viruses, HTLV-BLV complex, lentiviruses, and spumaviruses (Coffin, JM, Retroviridae: The viruses and their replication, In Fundamental Virology, Third Edition, BN Fields et al., Eds., Lippincott-Raven Publishers, Philadelphia, 1996). In some embodiments, the vector is a plasmid. In some other embodiments, the vector is a viral vector.In some such embodiments, the viral vector is a lentiviral vector or a gamma-retroviral vector.

[0123] In yet another aspect, the present disclosure provides an isolated host cell comprising a nucleic acid, expression construct, or vector encoding an anti-Nectin-4 antibody or antigen-binding fragment thereof described herein. As used herein, the term "host" refers to a cell or microorganism that is targeted for genetic modification with a heterologous or exogenous nucleic acid molecule to produce a polypeptide of interest (e.g., an anti-Nectin-4 antibody or antigen-binding fragment thereof). In certain embodiments, the host cell may, if desired, already possess or be modified to contain other genetic modifications (e.g., the inclusion of a detectable marker) that confer desired properties related or unrelated to the biosynthesis of the heterologous or exogenous protein. Two or more heterologous or exogenous nucleic acid molecules can be introduced into the host cell as separate nucleic acid molecules, as multiple individually regulated genes, as a polycistronic nucleic acid molecule, as a single nucleic acid molecule encoding a fusion protein, or any combination thereof. When two or more exogenous nucleic acid molecules are introduced into a host cell, it is understood that the two or more exogenous nucleic acid molecules can be introduced as a single nucleic acid molecule (e.g., in a single vector) on separate vectors and can be integrated into a single site or multiple sites on the host chromosome. The number of heterologous nucleic acid molecules or protein activities referred to refers to the number of encoding nucleic acid molecules or protein activities, not the number of separate nucleic acid molecules introduced into the host cell. Methods for producing anti-nectin-4 antibodies

[0124] The anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure can be produced by any method known in the art for antibody production. As an example, the anti-Nectin-4 antibody or antigen-binding fragment can be produced by a method using an isolated nucleic acid sequence encoding the anti-Nectin-4 antibody or antigen-binding fragment, a vector and host cell containing the nucleic acid sequence, and recombinant techniques for producing the antibody or antigen-binding fragment. The nucleic acid sequence encoding the anti-Nectin-4 antibody or antigen-binding fragment can be isolated into a replicable DNA vector for further cloning or expression. DNA encoding the anti-Nectin-4 antibody or antigen-binding fragment can be easily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes capable of specifically binding to genes encoding the antibody heavy and light chains). Many vectors known in the art can be used as vectors. Vector components generally may include, but are not limited to, one or more of the following: a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.

[0125] Suitable host cells for cloning or expressing the DNA vectors herein may be prokaryotes, yeast, or higher eukaryotic cells as described herein. Suitable host cells for expressing glycosylated antibodies or antigen-binding fragments may be derived from multicellular organisms. Examples of invertebrate cells include, but are not limited to, plant cells and insect cells. Host cells used to produce antibodies or antigen-binding fragments can be cultured in various commercially available media. When using recombinant techniques, antibodies or antigen-binding fragments can be produced, for example, intracellularly, in the periplasmic space, or directly secreted into the medium. When antibodies or antigen-binding fragments are produced intracellularly, particulate debris, either host cells or lysed fragments, can be removed, for example, by centrifugation or ultrafiltration. When antibodies or antigen-binding fragments are secreted into the medium, the supernatant from such expression systems can be concentrated using commercially available protein concentration filters. A protease inhibitor, e.g., phenylmethylsuphonyl fluoride, can be included in any of the above steps to inhibit proteolysis, and an antibiotic can be included to prevent the growth of adventitious contaminants.

[0126] Anti-Nectin-4 antibody or antigen-binding fragment compositions prepared from cells can be purified using, for example, hydroxylapatite chromatography, gel electrophoresis, dialysis, and affinity chromatography. The suitability of Protein A as an affinity ligand may depend on the species and isotype of any immunoglobulin Fc domains that may be present in the antibody or antigen-binding fragment. Other protein purification techniques, such as fractionation on an ion exchange column, ethanol precipitation, reverse-phase HPLC, chromatography on silica, chromatography on heparin SEPHAROSE™, chromatography on anion or cation exchange resins (e.g., polyaspartic acid columns), chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation, can also be used to recover the antibody or antigen-binding fragment. After any preliminary purification steps, the mixture containing the antibody or antigen-binding fragment and contaminants can be subjected to low-pH hydrophobic interaction chromatography.Methods for humanizing antibodies include, for example, humanization using CDR grafting (Jones et al., Nature 15 321:522 (1986)) and variations thereof, such as "reshaping" (Verhoeyen, et al., 1988 Science 239:1534-1536; Riechmann, et al., 1988 Nature 332:323-337; Tempest, et al., Bio / Technol 1991 9:266-271), "hyperchimerization" (Queen, et al., 1989 Proc Natl Acad Sci USA 86:10029-10033; Co, et al., 1991 Proc Natl Acad Sci USA 88:2869-2873; Co, et al., 1992 J Immunol 148:1149-1154), and "veneering" (Mark, et al., BW Metcalf, BJ Dalton (Eds.) Cellular adhesion: molecular definition to therapeutic potential. Plenum Press, New York; 1994:291-312). Hyperhumanization (Tan, et al., 2002 J Immunol 169: 1119-25) is another modified humanization method that can be used to graft non-human CDRs onto human germline antibody sequences with similar CDR canonical structures.

[0127] In certain embodiments, the anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, or a conjugate thereof with a myeloid cell agonist is humanized. immunostimulatory compounds

[0128] The anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure is conjugated to an immunostimulatory compound (e.g., a TLR8 agonist), generally via a linker, to form an immunostimulatory conjugate. The anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure can be conjugated to one or more immunostimulatory compounds, generally about 1 to about 10 compounds per antibody or antigen-binding fragment thereof, and preferably about 2 to about 4 compounds per antibody or antigen-binding fragment thereof.

[0129] In some embodiments, the immunostimulatory compound activates human immune cells such as dendritic cells, macrophages, monocytes, myeloid-derived suppressor cells, NK cells, B cells, T cells, or a combination thereof. In some embodiments, the immunostimulatory compound is a myeloid cell agonist. A myeloid cell agonist is a compound that activates or stimulates an immune response by myeloid cells. For example, a myeloid cell agonist can stimulate an immune response by causing myeloid cells to release cytokines, which results in immune cell activation. Stimulation of an immune response by a myeloid cell agonist can be measured in vitro by co-culturing immune cells (e.g., peripheral blood mononuclear cells (PBMCs)) with cells targeted by the conjugate and measuring cytokine release, chemokine release, immune cell proliferation, upregulation of immune cell activation markers, ADCP, and / or ADCC. Exemplary assays are described in the Examples. ADCC can be measured by determining the percentage of target cells remaining in the co-culture after administration of the conjugate with target cells and PBMCs.

[0130] For example, the immunostimulatory compound can act at a toll-like receptor (TLR), a nucleotide oligomerization domain-like receptor (NOD), a RIG-I-like receptor (RLR), a c-type lectin receptor (CLR), or a cytosolic DNA sensor (CDS), or a combination thereof.

[0131] In some embodiments, the immunostimulatory compound comprises a ligand for one or more TLRs selected from the group consisting of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, and TLR10.

[0132] In some embodiments, the immunostimulatory compound is a myeloid cell agonist. In some embodiments, the myeloid cell agonist is a ligand of TLR2 selected from the group consisting of (a) a heat-killed bacterial product, preferably HKAL, HKEB, HKHP, HKLM, HKLP, HKLR, HKMF, HKPA, HKPG, or HKSA, HKSP, and (b) a cell wall component product, preferably LAM, LM, LPS, LIA, LIA, PGN, FSL, Pam2CSK4, Pam3CSK4, or zymosan.

[0133] In some embodiments, the myeloid cell agonist is a ligand of TLR3 selected from the group consisting of lintatolimod, poly-ICLC, RIBOXXON®, Apoxxim, RIBOXXIM®, IPH-33, MCT-465, MCT-475, and ND-1.1.

[0134] In some embodiments, the myeloid cell agonist is a ligand for TLR4 selected from the group consisting of LPS, MPLA, or pyrimido[5,4-b]indole, such as those described in International Publication No. WO2014 / 052828 (Regents of the University of California).

[0135] In some embodiments, the myeloid cell agonist is a ligand for TLR5 selected from the group consisting of FLA and flagellin.

[0136] In some embodiments, the myeloid cell agonist is a ligand for TLR6.

[0137] In certain embodiments, the myeloid cell agonist is a TLR7 agonist and / or a TLR8 agonist. In certain embodiments, the myeloid cell agonist is a TLR7 agonist. In certain embodiments, the myeloid cell agonist is a TLR8 agonist. In some embodiments, the myeloid cell agonist selectively agonizes TLR7 but not TLR8. In other embodiments, the myeloid cell agonist selectively agonizes TLR8 but not TLR7.

[0138] In certain embodiments, the myeloid cell agonist is a TLR7 agonist. In certain embodiments, the TLR7 agonist is an imidazoquinoline, imidazoquinoline amine, thiazoquinoline, aminoquinoline, aminoquinazoline, pyrido[3,2-d]pyrimidine-2,4-diamine, pyrimidine-2,4-diamine, 2-aminoimidazole, 1-alkyl-1H-benzimidazol-2-amine, tetrahydropyridopyrimidine, heteroarothiadiazide-2,2-dioxide, benzonaphthyridine, thieno[3,2-d]pyrimidine ... imidine, 4-amino-imidazoquinoline, imidazo-pyridinone, imidazo-pyrimidinone, purine, fused pyrimidine-lactam, imidazo[4,5-c]quinolin-4-amine, imidazo[4,5-c]quinoline, pyrimidine, benzazepine, imidazo-pyridine, pyrrolo-pyrimidine, 2-amino-quinazoline, guanosine analog, adenosine analog, thymidine homopolymer, ssRNA, CpG-A, polyG10, and polyG3. In certain embodiments, the TLR7 agonist is selected from the group consisting of imidazoquinolines, imidazoquinoline amines, thiazoquinolines, aminoquinolines, aminoquinazolines, pyrido[3,2-d]pyrimidine-2,4-diamines, pyrimidine-2,4-diamines, 2-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, heteroarothiadiazide-2,2-dioxides, benzonaphthyridines, thieno[3,2-d]pyrimidines, 4-aminoimidazoles, 1-alkyl-1H-benzimidazol-2-amines, tetrahydropyridopyrimidines, heteroaromatic azides, benzonaphthyridines, thieno[3,2-d]pyrimidines, 4-aminoimidazoles, 1 ... In some embodiments, the TLR7 agonist is selected from 2-amino-imidazoquinoline, imidazo-pyridinone, imidazo-pyrimidinone, purine, fused pyrimidine-lactam, imidazo[4,5-c]quinolin-4-amine, imidazo[4,5-c]quinoline, pyrimidine, benzazepine, imidazo-pyridine, pyrrolo-pyrimidine, and 2-amino-quinazoline, but is other than guanosine analogs, adenosine analogs, thymidine homopolymers, ssRNA, CpG-A, polyG10, and polyG3. In some embodiments, the TLR7 agonist is a non-naturally occurring compound.Examples of TLR7 modulators include GS-9620, GSK-2245035, imiquimod, resiquimod, DSR-6434, DSP-3025, IMO-4200, MCT-465, MEDI-9197, 3M-051, SB-9922, 3M-052, Limtop, TMX-30X, TMX-202, RG-7863, and RG-7795. TLR7 modulator compounds are disclosed in U.S. Patent Application Publication No. US2016 / 0168164 (Janssen, thieno[3,2-d]pyrimidine derivatives), U.S. Patent Application Publication No. US2015 / 0299194 (Roche, 4-amino-imidazoquinoline derivatives), U.S. Patent Application Publication No. US2011 / 0098248 (Gilead, Sciences, Imidazo-pyridinones, Imidazo-pyrimidinones, and Purine Derivatives), U.S. Patent Application Publication No. US2010 / 0143301 (Gilead Sciences, Fused Pyrimidine-Lactam Derivatives), and U.S. Patent Application Publication No. US20090047249 (Gilead Sciences, Purine Derivatives), which publications are incorporated herein by reference. Further examples of TLR7 modulators include those described in International Publication No. WO2018 / 009916 (Stanford University / Bolt Biotherapeutics, imidazo[4,5-c]quinolin-4-amine derivatives), International Publication No. WO2018 / 112108 (Bolt Biotherapeutics, imidazo[4,5-c]quinoline, pyrimidine, benzazepine, imidazo-pyridine, pyrrolo-pyrimidine, and purine derivatives), U.S. Patent Application Publication No. US2019 / 0055247 (Bristol-Myers Squibb Company, Inc., and / or its affiliates). Squibb, purine derivatives), International Publication No. WO2018 / 198091 (Novartis, pyrrolo-pyrimidine derivatives), U.S. Patent Application Publication No. US2017 / 0121421 (Novartis, pyrrolo-pyrimidine derivatives), U.S. Pat. No. 10,253,003 (Janssen, 2-amino-quinazoline derivatives), and U.S. Pat. No. 10,233,184 (Roche, imidazo-pyrimidinone derivatives), which publications are incorporated herein by reference.In some embodiments, the TLR7 agonist has an EC50 value of 500nM or less by a PBMC assay measuring TNF-alpha or IFN-alpha production. In some embodiments, the TLR7 agonist has an EC50 value of 100nM or less by a PBMC assay measuring TNF-alpha or IFN-alpha production. In some embodiments, the TLR7 agonist has an EC50 value of 50nM or less by a PBMC assay measuring TNF-alpha or IFN-alpha production. In some embodiments, the TLR7 agonist has an EC50 value of 10nM or less by a PBMC assay measuring TNF-alpha or IFN-alpha production.

[0139] In certain embodiments, the myeloid cell agonist is a TLR8 agonist. In certain embodiments, the TLR8 agonist is a benzazepine, an imidazoquinoline, a thiazoloquinoline, an aminoquinoline, an aminoquinazoline, a pyrido[3,2-d]pyrimidine-2,4-diamine, a pyrimidine-2,4-diamine, a 2-aminoimidazole, a 1-alkyl-1H-benzimidazol-2-amine, a tetrahydropyridopyrimidine, a pyrido[3,2-d]pyrimidine, a dihydropyrimidinylbenzazepinecarboxamide, a benzo[b]azepine, a benzazepinedicarboxamide derivative having a tertiary amide, a benzazepinedicarboxamide derivative having a secondary amide, a quinazoline. , pyrido[3,2-d]pyrimidines, diamino-pyrimidines, amino-quinazolines, heterocyclic-substituted 2-amino-quinazolines, diamino-pyrimidines, piperidino-pyrimidines, alkylamino-pyrimidines, 8-substituted benzazepines, amino-diazepines, amino-benzo-diazepines, amido-indoles, amido-benzimidazoles, phenylsulfonamides, dihydropteridinones, fused amino-pyrimidines, quinazolines, pyrido-pyrimidines, amino-substituted benzazepines, pyrrolo-pyridines, imidazo-pyridine derivatives, and amino-benzazepines, and is other than ssRNA. In some embodiments, the TLR8 agonist is a non-naturally occurring compound. Examples of TLR8 agonists include sergantolimod, motolimod, resiquimod, 3M-051, 3M-052, MCT-465, IMO-4200, VTX-763, and VTX-1463. Examples of TLR8 modulator compounds are disclosed in U.S. Patent Application Publication No. US2018 / 0086755 (Gilead, pyrido[3,2-d]pyrimidine derivatives), WO2017216054 (Roche, dihydropyrimidinylbenzazepinecarboxamide derivatives), WO2017 / 190669 (Shanghai De NovoPharmatech, benzo[b]azepine derivatives), WO 2016 / 142250 (Roche, benzazepine dicarboxamide derivatives), WO 2017 / 202704 (Roche, benzazepine dicarboxamide derivatives with tertiary amides), WO 2017 / 202703 (Roche, benzazepine dicarboxamide derivatives with secondary amides), U.S. Patent Application Publication No. US 2017 / 0071944 (Gilead, quinazoline and pyrido[3,2-d]pyrimidine derivatives), U.S. Patent Application Publication No. US 2014 / 0045849 (Janssen, diamino-pyrimidine derivatives), U.S. Patent Application Publication No. US2014 / 0073642 (Janssen, Amino-quinazoline Derivatives), WO2014 / 056953 (Janssen, Pyrrolo[3,2-d]pyrimidine Derivatives), WO2014 / 076221 (Janssen, Heterocycle-Substituted 2-Amino-quinazoline Derivatives), WO2014 / 128189 (Janssen, Diamino-pyrimidine Derivatives), US2014 / 0350031 (Janssen, Piperidino-pyrimidine Derivatives), WO2014 / 023813 (Janssen, Alkyl-aminopyrimidine Derivatives), US2008 / 0234251 (Array No. US2008 / 0306050 (Array Biopharma, Amino-diazepine Derivatives), U.S. Patent Application Publication No. US2010 / 0029585 (VentiRx Pharma, Amino-benzazepine Derivatives), U.S. Patent Application Publication No. US2011 / 0092485 (VentiRx Pharma, Amino-benzazepine Derivatives), U.S. Patent Application Publication No. US2011 / 0118235 (VentiRx Pharma, Amino-benzazepine Derivatives), U.S. Patent Application Publication No. US2012 / 0082658 (VentiRx Pharma, Amino-benzazepine VTX-378), U.S. Patent Application Publication No. US2012 / 0219615 (VentiRx Pharma), U.S. Patent Application Publication No. US2014 / 0066432 (VentiRxNo. US2014 / 0088085 (VentiRx Pharma, amino-benzazepine and amino-benzo-diazepine derivatives), U.S. Patent Application Publication No. US2014 / 0275167 (Novira Therapeutics, amide-indole and amide-benzimidazole derivatives), and U.S. Patent Application Publication No. US2013 / 0251673 (Novira Therapeutics, phenylsulfonamide derivatives), which publications are incorporated herein by reference. Further examples of TLR8 modulators include those disclosed in U.S. Patent Application Publication No. US2016 / 0108045 (Gilead, dihydropteridinone derivatives), U.S. Patent Application Publication No. US2018 / 0065938 (Gilead, fused amino-pyrimidine derivatives), U.S. Patent Application Publication No. US2018 / 0263985 (Gilead, quinazoline and pyrido-pyrimidine derivatives), WO2017 / 046112 (Roche, amino-substituted benzazepine derivatives), WO2016 / 096778 (Roche, amino-substituted benzazepine derivatives), U.S. Patent Application Publication No. 2019 / 0016808 (Birdie In some embodiments, the TLR8 agonist has the structure: [ka] wherein the structure is optionally substituted at any position except the -NH2 position. In some embodiments, the TLR8 agonist has an EC50 value of 500 nM or less by a PBMC assay measuring TFN-alpha production. In some embodiments, the TLR8 agonist has an EC50 value of 100 nM or less by a PBMC assay measuring TFN-alpha production. In some embodiments, the TLR8 agonist has an EC50 value of 50 nM or less by a PBMC assay measuring TFN-alpha production. In some embodiments, the TLR8 agonist has an EC50 value of 10 nM or less by a PBMC assay measuring TFN-alpha production.

[0140] In some embodiments, the TLR8 agonist is a benzazepine selected from the compounds provided herein (eg, the Category A and Category C compounds).

[0141] In some embodiments, the myeloid cell agonist is a ligand of TLR9 selected from the group consisting of ODN1585, ODN1668, ODN1826, PF-3512676 (ODN2006), ODN2007, ODN2216, ODN2336, ODN2395, BB-001, BB-006, CYT-003, IMO-2055, IMO-2125, IMO-3100, IMO-8400, IR-103, IMO-9200, agatolimod, DIMS-9054, DV-1079, DV-1179, AZD-1419, leftolimod (MGN-1703), ritenimod, and CYT-003-QbGl0.

[0142] In other embodiments, the myeloid agonist selectively agonizes TLR9, TLR3, TLR4, TLR2, TLR5, RIG-I, STING, cGAS, NOD1, NOD2, NOD1 / NOD2, NRLP3, ALPK1, MDA5 AIM2, IRE1, and PERK.

[0143] In some embodiments, the myeloid cell agonist is a ligand for TLR10.

[0144] In some embodiments, the myeloid cell agonist is a nucleotide oligomerization domain (NOD)-like ligand selected from the group consisting of NOD1 agonists (C12-iE-DAP, iE-DAP, Tri-DAP), NOD2 agonists (L18-MDP, MDP, M-TriLYS, M-TriLYS-D-ASN, murabutide, N-Glycolyl-MDP), and NOD1 / NOD2 agonists (M-TriDAP, PGN).

[0145] In some embodiments, the myeloid cell agonist is a ligand of one or more RIG-I-like receptors (RLRs) selected from the group consisting of S'ppp-dsRNA, poly(dA:dT), poly(dG:dC), and poly(I:C).

[0146] In some embodiments, the myeloid cell agonist is a ligand for one or more C-type lectin receptors (CLRs) selected from the group consisting of Cnrdlan AL, HKCA, HKSC, WGP, zymosan, and trehalose-6,6-dibehenate.

[0147] In some embodiments, the myeloid cell agonist is selected from the group consisting of ADU-S100, c-GMP, cG-AMP, cG-GMP, cA-AMP, c-di-AMP, c-di-IMP, c-di-GMP, c-di-UMP, HSV-60, ISD, pCpG, poly(dA:dT), poly(dG:dC), poly(dA), VACV-70, and alpha-mangostin, and the compounds described in WO 2018 / 156625 (U of Texas), International Publication No. WO2018 / 152453 (Eisai), International Publication No. WO2018 / 138685 (Janssen), International Publication No. WO2018 / 100558 (Takeda), International Publication No. WO2018 / 098203 (Janssen), International Publication No. WO2018 / 065360 (Biolog Life Sciences), International Publication No. WO2018 / 060323 (Boehringer Ingelheim), International Publication No. WO2018 / 045204 (IFM Therapeutics), International Publication No. WO2018 / 009466 (Aduro), International Publication No. WO2017 / 161349 (Immune Sensor), WO2017 / 123669, WO2017 / 123657, WO2017 / 027646 (Merck), WO2017 / 027645 (Merck), WO2016 / 120305 (GSK), WO2016 / 096174 (InvivoGen), and U.S. Patent Application Publication No. US2014 / 0341976 (Aduro).

[0148] In some embodiments, the myeloid cell agonist is a ligand of an inflammasome inducer selected from the group consisting of (a) the NLRP3 inflammasome protein complex, preferably alum crystals, ATP, CPPD crystals, hemozoin, MSU crystals, nano-Si 02, nigericin, and (b) the AIM2 inflammasome protein complex, e.g., poly(dA:dT).

[0149] In certain embodiments, the TLR8 agonist is selected from Category A or Category C, or the TLR7 agonist is selected from Category B, as further described herein. The variables and formulas for Category A compounds (TLR8 agonists) are set forth in the section entitled Compounds of Category A, the variables and formulas for Category B compounds (TLR7 agonists) are set forth in the section entitled Compounds of Category B, and the variables and formulas for Category C compounds (TLR8 agonists) are set forth in the section entitled Compounds of Category C. The formulas and variables for Category A compounds, Category B compounds, and Category C compounds may overlap in nomenclature, e.g., Formula IA for each of Category A, Category B, and Category C compounds; however, the variable and formula descriptions are not intended to be interchangeable between categories. Category A compounds, TLR8 agonists

[0150] In some aspects, the myeloid cell agonist is a benzazepine-4-carboxamide compound. In certain embodiments, the benzazepine-4-carboxamide compound has the structure of formula X-1: [ka] (In the formula: R 1 is C 3~7 is alkyl; R 2 is C 3~7 Alkyl or C 3~7 Cycloalkyl-C 1~7 is alkyl; R 3 is hydrogen; R 4 teeth, C 1~7 Alkyl (C 1~7alkyl is unsubstituted or substituted with one or two groups selected from the group consisting of phenyl and heteroaryl, where the heteroaryl is an aromatic 5- or 6-membered ring containing 1, 2, or 3 atoms selected from nitrogen, oxygen, and / or sulfur; C 3~7 Cycloalkyl (C 3~7 Cycloalkyl is unsubstituted or is phenyl or phenylamino-C 1~4 substituted by alkyl), and Heterocyclyl (the heterocyclyl is a saturated 3- to 7-membered ring containing one heteroatom selected from N and O, and is unsubstituted or substituted by phenyl) selected from the group consisting of It has.

[0151] The structure of formula X-1 is described, for example, in International Publication No. WO2017 / 202703.

[0152] In some aspects, the myeloid cell agonist is a benzazepine-dicarboxamide compound. In certain embodiments, the benzazepine-dicarboxamide compound has the structure of formula X-2: [ka] (In the formula: R 1 is C 3~7 is alkyl; R 2 is C 3~7 Alkyl or C 3~7 Cycloalkyl-C 1~7 is alkyl; R 3 is (a) [ka] (In the formula, X1 is (CH2) m (wherein m is 1 or 2); X2 is (CH2) n (wherein n is 1 or 2); X3 is (CH2) o wherein o is 1 or 2; X4 is (CH2) p where p is 1 or 2; Z1 is phenyl, where phenyl is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, and di-C 1~7 Alkyl-amino-C 1~7 substituted with one or two groups selected from the group consisting of alkyl; or (b) [ka] (In the formula, X5 is (CH2) q (wherein q is 1 or 2); X6 is (CH2) r wherein r is 1 or 2; Y1 is a carbon or nitrogen atom; Z2 is hydrogen; Z3 is hydrogen, C 1~7 Alkoxy, C 2~7 Alkenyloxy, phenyl, phenyl-C 1~7 Alkyl, Phenyl-C 1~7 Alkyloxy, Phenyl-C 1~7 Alkylamino, phenylamino-C 1~7 alkyl, phenylamino, wherein phenyl is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, and di-C 1~7 Alkyl-amino-C 1~7 substituted with one or two groups selected from the group consisting of alkyl; or (c) [ka] (In the formula, X7 is (CH2) s (wherein s is 1 or 2); Z4 is phenyl, where phenyl is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, and di-C 1~7 Alkyl-amino-C 1~7 substituted with one or two groups selected from the group consisting of alkyl; or (d) [ka] (In the formula, X8 is (CH2) t (wherein t is 1 or 2); Z5 is phenyl, where phenyl is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, and di-C 1~7 Alkyl-amino-C 1~7substituted with one or two groups selected from the group consisting of alkyl is a heterocycle selected from It has.

[0153] Compounds of formula X-2 are described, for example, in International Publication No. WO2017 / 202704.

[0154] In some aspects, the myeloid cell agonist is a benzazepine sulfonamide compound. In certain embodiments, the benzazepine sulfonamide compound has the structure of formula X-3: [ka] (In the formula, R 1 and R 2 are the same or different, C 1~7 Alkyl, Hydroxy-C 2~7 Alkyl, Amino-C 2~7 Alkyl, C 2~7 Alkenyl, and C 3~7 alkynyl; R 3 is hydrogen or C 1~7 is alkyl; R 6 is hydrogen or C 1~7 is alkyl; R 4 and R 5 One of them is hydrogen, C 1~7 Alkyl, halogen-C 1~7 Alkyl, and C 1~7 alkoxy; R 4 and R 5 The other of [ka] (In the formula, R 7 and R 8 are the same or different, hydrogen, C 1~7 Alkyl, halogen-C 1~7 Alkyl, Hydroxy-C1~7 Alkyl, Hydroxy-C 1~7 Alkoxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, Amino-C 1~7 Alkoxy-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkoxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl-carbonyl, and C 1~7 Alkyl-amino(xamino)-C 1~7 alkyl-carbonyl; or R 7 and R 8 together with the nitrogen atom to which they are attached form a 4- to 6-membered heterocycle which is unsubstituted or 1~7 Alkyl-amino, hydroxy, and hydroxy-C 1~7 alkyl, and an additional NR 10 group (in the formula, R 10 is hydrogen, amino-C 1~7 Alkyl, and C 1~7 Alkyl-amino-C 1~7 alkyl); Y is N or CR 9 is; (In the formula, R 9 is hydrogen, C 1~7 Alkyl, and halogen-C 1~7 alkyl) It has.

[0155] Compounds of formula X-3 are described, for example, in International Publication No. WO2016 / 096778.

[0156] In some embodiments, the myeloid cell agonist is a dihydropyrimidinyl benzazepine carboxamide compound. In some embodiments, the dihydropyrimidinyl benzazepine carboxamide compound has the structure of formula X-4: [ka] (In the formula, R 1 is C 3~7 is alkyl; R 2 is C 3~7 Alkyl or C 3~7 Cycloalkyl-C 1~7 is alkyl; R 3 is hydrogen or C 1~7 is alkyl; R 4 is hydrogen or C 1~7 is alkyl; R 5 is hydrogen, halogen, C 1~7 Alkyl, and C 1~7 selected from the group consisting of alkoxy; R 6 is hydrogen, halogen, C 1~7 Alkyl, and C 1~7 selected from the group consisting of alkoxy; X is N or CR 7 (In the formula, R 7 is hydrogen, halogen, C 1~7 Alkyl, and C 1~7 alkoxy) It has.

[0157] Compounds of formula X-4 are described, for example, in International Publication No. WO2017 / 216054.

[0158] In some embodiments, the myeloid cell agonist is a sulfinylphenyl or sulfonimidoylphenyl benzazepine compound. In some embodiments, the sulfinylphenyl or sulfonimidoylphenyl benzazepine compound has the structure of Formula X-5: [ka] (In the formula, X is CR 7 or N; R 1 is C 3~7 Alkyl or C 3~7 is cycloalkyl; R 2 is C 3~7 Alkyl, Hydroxy-C 1~7 Alkyl, C 3~7 -Alkynyl, amino-C 1~7 Alkoxy-C 1~7 Alkoxy-C 1~7 Alkyl, halogen-C 1~7 Alkyl, and C 3~7 Cycloalkyl-C 1~7 selected from the group consisting of alkyl; R 3 and R 4 One of them is [ka] and R 3 and R 4 The other is hydrogen, C 1~7 selected from the group consisting of alkyl, and halogen; R 5 , R 6 , and R 7 are, independently of each other, hydrogen, C 1~7 alkyl, and halogen; R 8 is C 1~7 is alkyl; R 9 does not exist or =NR 10 (In the formula, R 10 is hydrogen, C 1~7 Alkyl, halogen-C 1~7 Alkyl, Hydroxy-C 1~7 Alkyl, and hydroxy-C 1~7 Alkoxy-C 1~7 alkyl) It has.

[0159] Compounds of formula X-5 are described, for example, in International Publication No. WO2017 / 046112.

[0160] In some embodiments, the myeloid cell agonist has the structure of Formula X-6: [ka] (In the formula, [ka] (1) is a double bond or a single bond; [ka] (2) is a single bond or a double bond, and R1 is absent; R2 and R3 are independently selected from H and lower alkyl, or R2 and R3 are joined to form a saturated carbocyclic ring having 3 to 7 ring members; One of R7 and R8 is -NR f R g , [ka] and the other is hydrogen; where R f and R g is lower alkyl, or R f and R g together with the nitrogen to which they are attached form a saturated heterocyclic ring having 4 to 6 ring members; R4 is -NR c R d -OR 10 and; R c and R d is lower alkyl, where alkyl is optionally substituted with one or more -OH; R 10is alkyl, where alkyl is optionally substituted with one or more -OH; Z is C, [ka] (1) is a double bond or Z is N; [ka] (1) is a single bond; R a and R b is H, alkyl, alkenyl, alkynyl, and R e wherein alkyl is independently selected from one or more -OR 10 or R e with substitution as necessary; R e is selected from -NH2, -NH(alkyl), and -N(alkyl)2; R 1 teeth, [ka] (2) is a double bond, it does not exist, or [ka] When (2) is a single bond, R 1 and R a or R b together with the atoms to which they are attached form a saturated, partially unsaturated, or unsaturated heterocyclic ring having 5 to 7 ring members, and R a or R b the other is hydrogen or is optionally absent to correspond to ring unsaturation. and a TLR modulator compound having the formula:

[0161] In some embodiments, the myeloid cell agonist has the structure of Formula X-7: [ka] (In the formula, Y is CF2CF3, CF2CF7R 6 or an aryl or heteroaryl ring, wherein the aryl and heteroaryl rings are alkenyl, alkynyl, Br, CN, OH, NR 6 R 7 , C(=O)R 8 , N.R. 6 SO2R 7 , (C1-C6 alkyl)amino, R 6 OC(=O)CH=CH2-, SR 6 , and SO2R 6 and wherein the aryl and heteroaryl rings are optionally further substituted with one or more groups independently selected from F, Cl, CF3, CF3O—, HCF2O—, alkyl, heteroalkyl, and ArO—; R 1 , R 3 and R 4 are independently selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl, where alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are independently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 or R 3 and R4 together with the atoms to which they are attached form a saturated or partially unsaturated carbocyclic ring, wherein the carbocyclic ring is selected from the group consisting of alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 and optionally substituted with one or more groups independently selected from: R 2 and R 8 H, OR 6 , N.R. 6 R 7 , alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl, wherein alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are independently selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 and optionally substituted with one or more groups independently selected from: R 5a , R5b , and R 5c are independently H, F, Cl, Br, I, OMe, CH3, CH2F, CHF2, or CF3; R 6 and R 7 are independently selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 or R 6 and R 7 together with the atoms to which they are attached form a saturated or partially unsaturated heterocyclic ring, wherein said heterocyclic ring is selected from the group consisting of alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R6 and optionally substituted with one or more groups independently selected from and a TLR modulator compound having the formula:

[0162] In some embodiments, the myeloid cell agonist has the structure of Formula X-8: [ka] (In the formula, W is -C(O)-; Z is H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, OR 6 , or NR 6 R 7 wherein alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl are alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OCC=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 and optionally substituted with one or more groups independently selected from: R 1 , R 2 , R 3 and R 4are independently selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(C=O)CH=CH2-, NR 6 SO2R 7 , SR6, and SO2R 6 or R 1 and R 2 together with the atoms to which they are attached form a saturated or partially unsaturated carbocyclic ring, wherein said carbocyclic ring is selected from the group consisting of alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 or R 3 and R 4 together are oxo; R 5is H, F, Cl, Br, I, OMe, CH3, CH2F, CHF2, CF3, or CF2CF3; R 6 and R 7 are independently selected from H, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl, wherein said alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, and heteroaryl are selected from alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6 optionally substituted with one or more groups independently selected from R 6 and R 7 together with the atoms to which they are attached form a saturated or partially unsaturated heterocyclic ring, wherein said heterocyclic ring is selected from the group consisting of alkyl, alkenyl, alkynyl, F, Cl, Br, I, CN, OR 6 , N.R. 6 R 7 , C(=O)R 6 , C(=O)OR 6 , OC(=O)R 6 , C(=O)NR 6 R 7 , (C1-C6 alkyl)amino, CH3OCH2O-, R 6 OC(=O)CH=CH2-, NR 6 SO2R 7 , S.R. 6 , and SO2R 6and optionally substituted with one or more groups independently selected from: n is 0, 1, 2, 3, or 4) and a TLR modulator compound having the formula:

[0163] Compounds of formula X-6, X-7, and X-8 are described, for example, in U.S. Publication Nos. US2019 / 0016808 and US2014 / 0088085.

[0164] In some embodiments, the myeloid cell agonist has the structure of formula X-9: [ka] (In the formula, R 1 is C 3~7 Alkyl or C 3~7 is cycloalkyl; R 2 is C 1~7 Alkyl, Hydroxy-C 1~7 Alkyl, C 2~7 Alkenyl, C 3~7 Alkynyl, Amino-C 1~7 Alkoxy-C 1~7 Alkyl, Amino-C 1~7 Alkoxy-C 1~7 Alkoxy-C 1~7 Alkyl, halogen-C 1~7 Alkyl, C 3~7 Cycloalkyl-C 1~7 Alkyl, and phenyl-C 1~7 alkyl, wherein phenyl is unsubstituted or selected from the group consisting of amino-C 1~7 substituted by alkyl; R 3 is hydrogen; R 4 teeth, Phenyl (the phenyl is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, Di-C 1~7 Alkyl-amino-C 1~7 Alkyl, Amino-C 2~7 Alkenyl, C 1~7 Alkyl-amino-C 2~7 Alkenyl, Di-C 1~7 Alkyl-amino-C 2~7 Alkenyl, Amino-C 2~7 Alkynyl, C 1~7 Alkyl-amino-C 2~7 Alkynyl, Di-C 1~7 Alkyl-amino-C 2~7 Alkynyl, benzyloxycarbonylamino-C 1~7 Alkyl, Amino-C 1~7 Alkoxy, Amino-C 1~7 Alkoxy-C 1~7 Alkoxy, Amino-C 1~7 Alkoxy-C 1~7 Alkyl, Amino-C 1~7 Alkoxy-C 1~7 Alkoxy-C 1~7 Alkyl, C 1~7 Alkyl sulfonyl, heterocyclyl carbonyl, and phenyl-C 1~7 alkyl, wherein the phenyl is unsubstituted or C 1~7 Alkoxy or Amino-C 1~7 substituted by alkyl), or Heteroaryl, wherein the heteroaryl is a 5- or 6-membered aromatic ring containing 1, 2, or 3 heteroatoms selected from N, O, or S, and is unsubstituted or C 1~7 Alkyl, halogen, halogen-C 1~7 Alkyl, C 1~7 Alkoxy, Hydroxy-C 1~7 Alkyl, Amino-C 1~7 Alkyl, C 1~7 Alkyl-amino-C 1~7 Alkyl, Di-C 1~7Alkyl-amino-C 1~7 Alkyl, Amino-C 2~7 Alkenyl, C 1~7 Alkyl-amino-C 2~7 Alkenyl, Di-C 1~7 Alkyl-amino-C 2~7 Alkenyl, Amino-C 2~7 Alkynyl, C 1~7 Alkyl-amino-C 2~7 Alkynyl, Di-C 1~7 Alkyl-amino-C 2~7 Alkynyl, benzyloxycarbonylamino-C 1~7 Alkyl, Amino-C 1~7 Alkoxy, Amino-C 1~7 Alkoxy-C 1~7 Alkoxy, Amino-C 1~7 Alkoxy-C 1~7 Alkyl, Amino-C 1~7 Alkoxy-C 1~7 Alkoxy-C 1~7 Alkyl, C 1~7 Alkyl sulfonyl, heterocyclyl carbonyl, and phenyl-C 1~7 alkyl, wherein the phenyl is unsubstituted or C 1~7 Alkoxy or Amino-C 1~7 (substituted by alkyl) selected from the group consisting of and a TLR modulator compound having the formula:

[0165] Compounds of formula X-9 are described, for example, in International Publication No. WO2016 / 142250.

[0166] In some aspects, the present disclosure provides a compound having the structure of formula (IIA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: [ka] represents an optional double bond; L 10 -X 10 - and; L 2 -X 2 -, -X 2 -C 1~6 Alkylene-X 2 -, -X 2 -C 2~6 Alkenylene-X 2 - and -X 2 -C 2~6 Alkynylene-X 2 -, each of which in alkylene, alkenylene, or alkynylene is selected from one or more R 12 with substitution as necessary; X 10 -C(O)- and -C(O)N(R 10 )- * (In the formula, * is X 10 R 5 (representing the point of attachment to the X 2 In each occurrence, -O-, -S-, -N(R 10 )-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 ), -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10)-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O), -OS(O)2-, -S(O)2O, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, and -N(R 10 )S(O)N(R 10 )-independently selected from; R 1 and R 2 is hydrogen; and C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and —CN); R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10)C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 selected from carbocycles and 3- to 12-membered heterocycles, where R 4 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 5 is unsaturated C 4~8 carbocycle; bicyclic carbocycle; and fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle, wherein R 5 is optionally substituted, and the substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10, -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 independently selected from carbocycle and 3- to 12-membered heterocycle, wherein R 5 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 10 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, -C 1~10 Haloalkyl, -OC 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10)2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 independently selected from carbocycle and 3- to 10-membered heterocycle, wherein R 12 Each C in 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings are not substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; Any substitutable carbon on the benzazepine core can be R 12 or two substituents on a single carbon atom may be combined to form a 3- to 7-membered carbocyclic ring. The present invention provides a TLR8 agonist represented by

[0167] In some embodiments, the compound of formula (IIA) has formula (IIB): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 20 , R 21 , R 22 , and R 23is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 independently selected from alkynyl; R 24 and R 25 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl; or R 24 and R 25 are taken together and optionally substituted saturated C 3~7 forming a carbocyclic ring) is expressed by

[0168] In some embodiments, R 20 , R 21 , R 22 , and R 23 is hydrogen, halogen, -OH, -OR 10 , -NO2, -CN, and C 1~10 R is independently selected from alkyl. 20 , R 21 , R 22 , and R 23 and R may each be hydrogen. 21is halogen. In certain embodiments, R 21 is hydrogen. In certain embodiments, R 21 -OR 10 For example, R 21 may be —OCH3.

[0169] In some embodiments, R 24 and R 25 are hydrogen, halogens, -OH, -NO2, -CN, and C 1~10 alkyl, or R 24 and R 25 are taken together and optionally substituted saturated C 3~7 In certain embodiments, R 24 and R 25 are each hydrogen. In another embodiment, R 24 and R 25 are taken together and optionally substituted saturated C 3~5 forming a carbocyclic ring, where the substituents are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 carbocycle, and 3- to 12-membered heterocycle, each of which is independently optionally substituted with one or more substituents independently selected from:

[0170] In some embodiments, R 1 is hydrogen. In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is -C(O)-.

[0171] In some embodiments, L 10 is -C(O)N(R 10 )- * In certain embodiments, —C(O)N(R 10 )- * R 10 is hydrogen and C 1~6 For example, L 10 is -C(O)NH- * may be.

[0172] In some embodiments, R 5 is an optionally substituted bicyclic carbocycle. In certain embodiments, R 5 R is an optionally substituted 8- to 12-membered bicyclic carbocycle. 5 is halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 and optionally substituted 8-12 membered bicyclic carbocyclic ring substituted with one or more substituents independently selected from alkynyl. 5-OR 10 , -N(R 10 )2, and ═O. In some embodiments, R 5 is an optionally substituted indane and an optionally substituted tetrahydronaphthalene. 5 teeth, [ka] any one of which may be optionally substituted. For example, R 5 teeth, [ka] is selected from.

[0173] In some embodiments, R 5 is an optionally substituted unsaturated C 4~8 In certain embodiments, R 5 is an optionally substituted unsaturated C 4~6 In certain embodiments, R 5 is replaced by C 3~12 an optionally substituted unsaturated C ring having one or more substituents independently selected from a carbocycle and an optionally substituted 3- to 12-membered heterocycle; 4~6 It is a carbocyclic ring. 5 is an optionally substituted phenyl, an optionally substituted 3- to 12-membered heterocycle, an optionally substituted C 1~10 Alkyl, optionally substituted C 2~10 an optionally substituted unsaturated C having one or more substituents independently selected from alkenyl, and halogen; 4~6 It may also be a carbocyclic ring.

[0174] In some embodiments, R 5 is selected from optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycles.5 is -C(O)OR 10 , -N(R 10 )2, -OR 10 , and optionally substituted C 1~10 and optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycles having one or more substituents independently selected from alkyl. 5 is -C(O)OR 10 In certain embodiments, R is an optionally substituted fused 5-5, fused 5-6, and fused 6-6 bicyclic heterocycle substituted with 5 is an optionally substituted fused 6-6 bicyclic heterocycle. For example, the fused 6-6 bicyclic heterocycle may be an optionally substituted pyridine-piperidine. In some embodiments, L 10 is attached to the carbon atom of the pyridine of the fused pyridine-piperidine. 5 is selected from tetrahydroquinoline, tetrahydroisoquinoline, tetrahydronaphthyridine, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted. 5 may be an optionally substituted tetrahydronaphthyridine. In some embodiments, R 5 teeth, [ka] [ka] [ka] is selected from.

[0175] In some embodiments, R 5 If is substituted, R 5 The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from alkynyl. 5The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from 5 The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, and -CN; and halogens, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R10 , -C(O)OR 10 C optionally substituted with one or more substituents independently selected from -NO, -O, and -CN. 1~10 In some embodiments, R 5 is not substituted.

[0176] In some embodiments, L 2 is -C(O)-, and -C(O)NR 10 In some embodiments, L 2 is —C(O)—. In some embodiments, L 2 is -C(O)NR 10 -C(O)NR 10 -R 10 is hydrogen and C 1~6 For example, L 2 may be —C(O)NH—.

[0177] In some embodiments, R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl). 4 -OR 10 , and -N(R 10 )2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocyclic rings, and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl). 4 is -N(R 10 )2. -N(R 10 )2R10 is replaced by C in each occurrence as needed. 1~6 In certain embodiments, —N(R 10 )2R 10 is, at each occurrence, independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted. For example, R 4 teeth, [ka] In certain embodiments, -L 2 -R 4 teeth, [ka] is.

[0178] In some embodiments, R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from alkynyl. 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10)2, -NO2, =O, =S, =N(R 10 ), and -CN; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 carbocycle, and 3- to 10-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0179] In some embodiments, the compound of formula (IIB) is a compound of formula (IIC): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 1 and R 2 is hydrogen; L 2 is -C(O)-; R 4 is -N(R 10 )2; R 10 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, -C 1~10 Haloalkyl, -OC 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; L 10 is -C(O)N(R 10 )- * (In the formula, * L 10 R 5 ) and R 5 is a fused 5-5, fused 5-6, or fused 6-6 bicyclic heterocycle, where R 5 is optionally substituted, and the substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10)2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic rings, and 3- to 12-membered heterocyclic rings (each of which may contain halogen, OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from is.

[0180] In certain embodiments, —N(R 10 )2R 10 is, at each occurrence, independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted; and / or —C(O)N(R 10 )- * R 10 is hydrogen.

[0181] In certain embodiments, R 4 teeth, [ka] and / or -C(O)N(R 10 )- * R10 is hydrogen.

[0182] In some embodiments, the compound is [ka] [ka] [ka] and any one of salts thereof.

[0183] In some aspects, the present disclosure provides a compound having the structure of formula (IIIA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: [ka] represents an optional double bond; L 11 -X 11 - and; L 2 -X 2 -, -X 2 -C 1~6 Alkylene-X 2 -, -X 2 -C 2~6 Alkenylene-X 2 - and -X 2 -C 2~6 Alkynylene-X 2 -, each of which in alkylene, alkenylene, or alkynylene is selected from one or more R 12 with substitution as necessary; X 11 is -C(O)- and -C(O)N(R 10 )- * (In the formula, * is X 11 R 6 (representing the point of attachment to the X 2 In each occurrence, -O-, -S-, -N(R 10 )-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, -S(O)2O-, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, and -N(R 10 )S(O)N(R 10 )-independently selected from; R 1 and R 2 is hydrogen; C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10, -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and —CN); R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 selected from carbocycles and 3- to 12-membered heterocycles, where R 4 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 6 is selected from phenyl and 5- or 6-membered heteroaryl, any one of which is R 7 and R 6 and R 12 and optionally further substituted by one or more additional substituents independently selected from: R 7 -C(O)NHNH2, -C(O)NH-C 1~3 Alkylene-NH(R 10 ), -C(O)CH3, -C 1~3 Alkylene-NHC(O)OR 11 , -C 1~3 Alkylene-NHC(O)R 10 , -C 1~3 Alkylene-NHC(O)NHR 10 , -C 1~3 Alkylene-NHC(O)-C 1~3 Alkylene-R 10 , and R 12 a 3- to 12-membered heterocycle optionally substituted with one or more substituents independently selected from: R 10 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, -C 1~10 Alkyl, -C 1~10 Haloalkyl, -OC 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C3~12 carbocycle, and 3- to 12-membered heterocycle; R 11 is C 3~12 carbocycle and 3- to 12-membered heterocycle, each of which is selected from R 12 and optionally substituted with one or more substituents independently selected from: R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C3~10 independently selected from carbocycle and 3- to 10-membered heterocycle, wherein R 12 Each C in 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings are not substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; Any substitutable carbon on the benzazepine core can be R 12 or two substituents on a single carbon atom may be combined to form a 3- to 7-membered carbocyclic ring. The present invention provides a compound represented by the formula:

[0184] In some embodiments, the compound of formula (IIIA) has formula (IIIB): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 20 , R 21 , R 22 , and R 23 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 independently selected from alkynyl; R 24 and R 25 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl; or R 24 and R 25 are taken together and optionally substituted saturated C 3~7 forming a carbocyclic ring) is expressed by

[0185] In some embodiments, R 20 , R 21 , R 22 , and R 23 are hydrogen, halogens, -OH, -NO2, -CN, and C 1~10 In certain embodiments, R 20 , R 21 , R 22 , and R 23 are each hydrogen. 24 and R 25 are hydrogen, halogens, -OH, -NO2, -CN, and C 1~10 alkyl, or R 24 and R 25 are taken together and optionally substituted saturated C3~7 In certain embodiments, R 24 and R 25 are each hydrogen. In certain embodiments, R 24 and R 25 are taken together and optionally substituted saturated C 3~5 Forms a carbocyclic ring.

[0186] In some embodiments, R 1 is hydrogen. In some embodiments, R 2 is hydrogen.

[0187] In some embodiments, L 11 is -C(O)N(R 10 )- * In some embodiments, —C(O)N(R 10 )- * R 10 is hydrogen and C 1~6 For example, L 11 is -C(O)NH- * may be.

[0188] In some embodiments, R 6 is R 7 and R 6 is phenyl substituted with R 12 In some embodiments, R 6 -C(O)NHNH2, -C(O)NH-C 1~3 Alkylene-NH(R 10 ), -C 1~3 Alkylene-NHC(O)R 10 and -C(O)CH; and 3- to 12-membered heterocycles (which include -OH, -N(R 10 )2, -NHC(O)(R 10 ), -NHC(O)O(R 10 ), -NHC(O)N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -C(O)2R 10, and -C 1~3 Alkylene-(R 10 phenyl optionally substituted with one or more substituents independently selected from 6 is R 12 and optionally further substituted with one or more additional substituents independently selected from, for example, R 6 teeth, [ka] [ka] may be selected from:

[0189] In some embodiments, R 6 is R 7 and R is selected from 5- and 6-membered heteroaryl substituted with one or more substituents independently selected from 6 is R 12 In certain embodiments, R 6 is -C(O)CH3, -C 1~3 Alkylene-NHC(O)OR 10 , -C 1~3 Alkylene-NHC(O)R 10 , -C 1~3 Alkylene-NHC(O)NHR 10 , and -C 1~3 Alkylene-NHC(O)-C 1~3 Alkylene-(R 10 ); and 3- to 12-membered heterocycles (which include -OH, -N(R 10 )2, -NHC(O)(R 10 ), -NHC(O)O(R 10 ), -NHC(O)N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -C(O)2R 10 , and -C 1~3 Alkylene-(R 10), optionally substituted with one or more substituents independently selected from: 6 is R 12 and optionally further substituted with one or more additional substituents independently selected from R 6 may be selected from substituted pyridines, pyrazines, pyrimidines, pyridazines, furans, pyrans, oxazoles, thiazoles, imidazoles, pyrazoles, oxadiazoles, oxathiazoles, and triazoles; R 6 is R 12 In some embodiments, R 6 is a substituted pyridine, and R 6 is R 12 and optionally further substituted with one or more additional substituents independently selected from R 6 is as follows: [ka] [ka] In some embodiments, R 6 is a substituted pyridine, where R 7 -C 1~3 Alkylene-NHC(O)-C 1~3 Alkylene-R 10 In certain embodiments, R 7 is -C1 alkylene-NHC(O)-C1 alkylene-R 10 In certain embodiments, R 7 is -C1 alkylene-NHC(O)-C1 alkylene-NH2. In some embodiments, R 6 teeth, [ka] [ka] In certain embodiments, R 6 teeth, [ka] is.

[0190] In some embodiments, L 2 is -C(O)-, and -C(O)NR 10 In some embodiments, L 2 is -C(O)NR 10 -C(O)NR 10 -R 10 is hydrogen and C 1~6 For example, L 2 may be —C(O)NH—. In some embodiments, L 2 is -C(O)-.

[0191] In some embodiments, R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl). 4 -OR 10 and -N(R 10 )2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which, in each occurrence, may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 In certain embodiments, R is selected from the group consisting of alkynyl, alkynyl, and alkynyl. 4 is -N(R 10 )2. -N(R10 )2R 10 is replaced by C in each occurrence as needed. 1~6 In some embodiments, —N(R 10 )2R 10 is, at each occurrence, independently selected from methyl, ethyl, propyl, and butyl, any of which is optionally substituted. For example, R 4 teeth, [ka] In some embodiments, -L 2 -R 4 teeth, [ka] is.

[0192] In some embodiments, R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which, at each occurrence, is selected from halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 and optionally substituted independently with one or more substituents selected from a carbocycle, and a 3- to 10-membered heterocycle; 3~10 Carbocycles and 3- to 10-membered heterocycles (each of which, in each occurrence, may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl). 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10)2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which, at each occurrence, is selected from halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 carbocycle, and 3- to 10-membered heterocycle).

[0193] In some embodiments, the compound is [ka] [ka] [ka] [ka] [ka] and any one of salts thereof.

[0194] In some aspects, the present disclosure provides a compound having the structure of formula (IA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: [ka] represents an optional double bond; L 1 -X 1 -, -X 2 -C 1~6 Alkylene-X 2 -C 1~6 Alkylene-, -X 2 -C 2~6 Alkenylene-X 2 - and -X 2 -C 2~6 Alkynylene-X 2 -, each of which in alkylene, alkenylene, or alkynylene is selected from one or more R 12 with substitution as necessary; L 2 -X 2 -, -X 2 -C 1~6 Alkylene-X 2 -, -X 2 -C 2~6 Alkenylene-X 2 - and -X 2 -C 2~6 Alkynylene-X 2 -, each of which in alkylene, alkenylene, or alkynylene is selected from one or more R 12 with substitution as necessary; X 1 -S- * , -N(R 10 )- * , -C(O)O- * , -OC(O)- * , -OC(O)O- * , -C(O)N(R 10 )C(O)- * , -C(O)N(R 10 )C(O)N(R 10 ) * , -N(R 10 )C(O)-* , -CR 10 2N(R 10 )C(O)- * , -N(R 10 )C(O)N(R 10 )- * , -N(R 10 )C(O)O- * , -OC(O)N(R 10 )- * , -C(NR 10 )- * , -N(R 10 )C(NR 10 )- * , -C(NR 10 )N(R 10 )- * , -N(R 10 )C(NR 10 )N(R 10 )- * , -S(O)2- * , -OS(O)- * , -S(O)O- * , -S(O), -OS(O)2- * , -S(O)2O * , -N(R 10 )S(O)2- * , -S(O)2N(R 10 )- * , -N(R 10 )S(O)- * , -S(O)N(R 10 )- * , -N(R 10 )S(O)2N(R 10 )- * , and -N(R 10 )S(O)N(R 10 )- * (In the formula, * is X 1 R 3 (representing the point of attachment to the X 2 In each occurrence, -O-, -S-, -N(R 10 )-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R10 )C(O)N(R 10 ), -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O), -OS(O)2-, -S(O)2O, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, and -N(R 10 )S(O)N(R 10 )-independently selected from; R 1 and R 2 is hydrogen; C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and —CN); R 3 is replaced by C 3~12carbocycle, and optionally substituted 3- to 12-membered heterocycle, wherein R 3 The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 independently selected from carbocycle and 3- to 12-membered heterocycle, wherein R 3 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10, -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 selected from carbocycles and 3- to 12-membered heterocycles, where R 4 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are not subject to halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 10 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 independently selected from carbocycle and 3- to 10-membered heterocycle, wherein R 12 Each C in 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings are not substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; Any substitutable carbon on the benzazepine core can be R 12 or two substituents on a single carbon atom may be combined to form a 3- to 7-membered carbocyclic ring. The present invention provides a compound represented by the formula:

[0195] In some embodiments, the compound of formula (IA) has the formula (IB): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 20 , R 21 , R 22 , and R 23 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 independently selected from alkynyl; R 24 and R 25 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl; or R 24 and R 25 are taken together and optionally substituted saturated C 3~7 forming a carbocyclic ring) is expressed by

[0196] In some embodiments, R 20 , R21 , R 22 , and R 23 are hydrogen, halogens, -OH, -NO2, -CN, and C 1~10 In certain embodiments, R 20 , R 21 , R 22 , and R 23 are hydrogen atoms.

[0197] In some embodiments, R 24 and R 25 are hydrogen, halogens, -OH, -NO2, -CN, and C 1~10 alkyl, or R 24 and R 25 are taken together and optionally substituted saturated C 3~7 In some embodiments, R 24 and R 25 are each hydrogen. 24 and R 25 are taken together and optionally substituted saturated C 3~5 Forms a carbocyclic ring.

[0198] In some embodiments, R 1 is hydrogen. In some embodiments, R 2 is hydrogen.

[0199] In some embodiments, L 1 is -N(R 10 )C(O)- * , -S(O)2N(R 10 )- * , -CR 10 2N(R 10 )C(O)- * , and -X 2 -C 1~6 Alkylene-X 2 -C 1~6 In some embodiments, L is selected from alkylene-. 1 is -N(R 10 )C(O)- *In certain embodiments, —N(R 10 )C(O)- * R 10 is hydrogen and C 1~6 For example, L 1 is -NHC(O)- * In some embodiments, L 1 is -S(O)2N(R 10 )- * In certain embodiments, —S(O)N(R 10 )- * R 10 is hydrogen and C 1~6 For example, L 1 is -S(O)2NH- * In some embodiments, L 1 -CR 10 2N(R 10 )C(O)- * In certain embodiments, L 1 is -CH2N(H)C(O)- * and -CH(CH3)N(H)C(O)- * is selected from.

[0200] In some embodiments, R 3 is replaced by C 3~12 carbocycle, and optionally substituted 3- to 12-membered heterocycle, wherein R 3 The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from alkynyl. 3 is replaced by C 3~12 carbocycle, and optionally substituted 3- to 12-membered heterocycle, wherein R 3 The above substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0201] In some embodiments, R 3 is selected from optionally substituted aryl and optionally substituted heteroaryl. In some embodiments, R 3 is optionally substituted heteroaryl. 3 is halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 and optionally substituted heteroaryl, substituted with one or more substituents independently selected from alkynyl. In certain embodiments, R3 is selected from optionally substituted 6-membered heteroaryl. For example, R 3 may be an optionally substituted pyridine. In some embodiments, R 3 is optionally substituted aryl. In certain embodiments, R 3 is halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 R is an optionally substituted aryl substituted with one or more substituents independently selected from alkynyl. 3 may be an optionally substituted phenyl. In certain embodiments, R 3 R is selected from pyridine, phenyl, tetrahydronaphthalene, tetrahydroquinoline, tetrahydroisoquinoline, indane, cyclopropylbenzene, cyclopentapyridine, and dihydrobenzoxaborole, any one of which is optionally substituted. 3 teeth, [ka] [ka] any one of which may be optionally substituted. For example, R 3 teeth, [ka] [ka] may be selected from:

[0202] In some embodiments, L 2is -C(O)-, and -C(O)NR 10 In certain embodiments, L 2 is —C(O)—. In certain embodiments, L 2 is -C(O)NR 10 -C(O)NR 10 -R 10 is hydrogen and C 1~6 For example, L 2 may be —C(O)NH—.

[0203] In some embodiments, R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from:

[0204] In some embodiments, R 4 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from. In some embodiments, R 4 -OR 10 , and -N(R 10 )2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C3~12 Carbocyclic rings, and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl). 4 is -N(R 10 )2. -N(R 10 )2R 10 is replaced by C in each occurrence as needed. 1~6 In certain embodiments, —N(R 10 )2R 10 is, at each occurrence, independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted. For example, R 4 teeth, [ka] In certain embodiments, L 2 -R 4 teeth, [ka] is.

[0205] In some embodiments, R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6Alkyl, C 2~6 Alkenyl, C 2~6 alkynyl). 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 carbocycle, and 3- to 10-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0206] In some embodiments, the compound is [ka] and any one of salts thereof.

[0207] In some aspects, the present disclosure provides a compound having the structure of formula (IVA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: [ka] represents an optional double bond; L 12 -X 3 -, -X 3 -C 1~6 Alkylene-X 3 -, -X 3 -C 2~6 Alkenylene-X 3 - and -X 3 -C 2~6 Alkynylene-X 3 -, each of which in alkylene, alkenylene, or alkynylene is selected from R 12 and optionally substituted with one or more substituents independently selected from: L 22 -X 4 -, -X 4 -C 1~6 Alkylene-X 4 -, -X 4 -C 2~6 Alkenylene-X 4 - and -X 4 -C 2~6 Alkynylene-X 4 -, each of which in alkylene, alkenylene, or alkynylene is independently selected from R 10 and optionally substituted with one or more substituents independently selected from: X 3 and X 4 In each occurrence, -O-, -S-, -N(R 10)-, -C(O)-, -C(O)O-, -OC(O)-, -OC(O)O-, -C(O)N(R 10 )-, -C(O)N(R 10 )C(O)-, -C(O)N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)-, -N(R 10 )C(O)N(R 10 )-, -N(R 10 )C(O)O-, -OC(O)N(R 10 )-, -C(NR 10 )-, -N(R 10 )C(NR 10 )-, -C(NR 10 )N(R 10 )-, -N(R 10 )C(NR 10 )N(R 10 )-, -S(O)2-, -OS(O)-, -S(O)O-, -S(O)-, -OS(O)2-, -S(O)2O-, -N(R 10 )S(O)2-, -S(O)2N(R 10 )-, -N(R 10 )S(O)-, -S(O)N(R 10 )-, -N(R 10 )S(O)2N(R 10 )-, and -N(R 10 )S(O)N(R 10 )-independently selected from; R 1 and R 2 L 3 and hydrogen; and C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 Alkynyl (each of these is L 3 and each of these is optionally bonded to a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R10 , -NO2, =O, =S, =N(R 10 ), and —CN); R 4 and R 8 -OR 10 , -N(R 10 )2, -C(O)N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -S(O)R 10 , and -S(O)R 10 ;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of these is L 3 and each of these is optionally bonded to a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 independently selected from carbocycle and 3- to 12-membered heterocycle, wherein R 4 and R 8 Each C in 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings are represented by L 3 and optionally bonded to each of these, a halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10)2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; R 10 In each occurrence, L 3 , hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; L 3 is a linker moiety, where L 3 There is at least one occurrence of; R 12 In each occurrence, represents a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R10 ), and -CN;C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 3~10 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 10-membered heterocycle); C 3~10 independently selected from carbocycle and 3- to 10-membered heterocycle, wherein R 12 Each C in 3~10 Carbocyclic and 3- to 10-membered heterocyclic rings are not substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -C(O)R 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -S(O)R 10 , -S(O)2R 10 , -P(O)(OR 10 )2, -OP(O)(OR 10 )2, -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 optionally substituted with one or more substituents independently selected from alkynyl; Any substitutable carbon on the benzazepine core can be R12 or two substituents on a single carbon atom may be combined to form a 3- to 7-membered carbocyclic ring. The present invention provides a compound represented by the formula:

[0208] In some embodiments, the compound of formula (IVA) has formula (IVB): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 20 , R 21 , R 22 , and R 23 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 independently selected from alkynyl; R 24 , and R 25 is hydrogen, halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl; or R 24 and R 25are taken together and optionally substituted saturated C 3~7 forming a carbocyclic ring) is expressed by

[0209] In some embodiments, R 1 L 3 In some embodiments, R 2 L 3 is.

[0210] In some embodiments, L 12 is -C(O)N(R 10 )-. In some embodiments, -C(O)N(R 10 )-R 10 is hydrogen, C 1~6 Alkyl, and L 3 For example, L 12 may be —C(O)NH—.

[0211] In some embodiments, R 8 is an optionally substituted 5- or 6-membered heteroaryl. 8 L 3 In some embodiments, R 8 L 3 is an optionally substituted pyridine bonded to

[0212] In some embodiments, L 22 is -C(O)-, and -C(O)NR 10 In certain embodiments, L 22 is —C(O)—. In certain embodiments, L 22 is -C(O)NR 10 -C(O)NR 10 -R 10 is hydrogen, C 1~6 Alkyl, and -L 3 For example, L 22 may be —C(O)NH—.

[0213] In some embodiments, R 4 -OR 10 , and -N(R 10 )2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycle, 3- to 12-membered heterocycle, aryl, and heteroaryl (each of which may be substituted with halogen, -OR 10 , -SR 10 , -N(R 10 )2, -S(O)R 10 , -S(O)2R 10 , -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl, each of which is optionally substituted with one or more substituents independently selected from L 3 In some embodiments, R 4 is -N(R 10 )2 and -N(R 10 )2R 10 L 3 and hydrogen, wherein —N(R 10 )2 at least one R 10 L 3 is.

[0214] In some embodiments, the compound of formula (IVB) is a compound of formula (IVC): [ka] or a pharmaceutically acceptable salt thereof, (In the formula: R 1 and R 2 is hydrogen; L 22 is -C(O)-; R 4 -N(R10 )2; R 10 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, 3- to 12-membered heterocycle, and haloalkyl; L 12 is --C(O)N(R 10 )- * , (in the formula, * L 12 R 8 ) and R 8 is the linker moiety L 3 and optionally substituted 5-5, 5-6, or 6-6 bicyclic heterocycles bonded to, wherein the optional substituents, at each occurrence, are halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), and -CN; C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 Alkynyl (each of which is a halogen, -OR 10 , -SR10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 10 , -SR 10 , -C(O)N(R 10 )2, -N(R 10 )C(O)R 10 , -N(R 10 )C(O)N(R 10 )2, -N(R 10 )2, -C(O)R 10 , -C(O)OR 10 , -OC(O)R 10 , -NO2, =O, =S, =N(R 10 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, optionally substituted with one or more substituents independently selected from is.

[0215] In certain embodiments, —N(R 10 )2R 10 is, at each occurrence, independently selected from methyl, ethyl, propyl, and butyl, any one of which is optionally substituted. In certain embodiments, —C(O)N(R 10 )- * R 10 is hydrogen.

[0216] In some embodiments, the compound comprises a linker L 3 In some embodiments, L 3 is a non-cleavable linker. In some embodiments, L 3 is a cleavable linker. 3 may be cleavable by a lysosomal enzyme. In some embodiments, the compound is covalently attached to the antibody or antigen-binding fragment thereof.

[0217] In some embodiments, L 3 is the expression: [ka] (In the formula: L 4 represents the C-terminus of the peptide, and L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 32 and wherein R X is a reactive moiety; R 32 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 alkynyl (each of which is optionally substituted with one or more substituents independently selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, —NO) is expressed by

[0218] In some embodiments, Rx comprises a leaving group. In some embodiments, Rx comprises a maleimide. In some embodiments, L 3 is further covalently attached to an antibody or antigen-binding fragment thereof.

[0219] In some embodiments, L 3 is the expression: [ka] (In the formula, L 4 represents the C-terminus of the peptide, L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 32 and optionally substituted with one or more groups independently selected from: RX * comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached to a residue of an antibody or antigen-binding fragment thereof, RX * Above [ka] represents the point of attachment to a residue of an antibody or antigen-binding fragment thereof; R 32 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10 alkynyl (each of which is optionally substituted with one or more substituents independently selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, —NO) In some embodiments, L 3 The peptide contains Val-Cit or Val-Ala.

[0220] In some aspects, the present disclosure provides: [ka] [ka] [ka] [ka] and a salt of any one thereof. In some aspects, the present disclosure provides: [ka] [ka] [ka] [ka] [ka] and any one of the salts thereof (In the formula, RX * is a bond, succinimide moiety, or hydrolyzed succinimide moiety attached to a residue of an antibody or antigen-binding fragment thereof, RX * Above [ka] represents the point of attachment to a residue of the antibody or antigen-binding fragment thereof) The present invention provides a compound or salt selected from:

[0221] In some embodiments, L 3 is the expression: [ka] (wherein Rx comprises a reactive moiety and n=0 to 9) In some embodiments, Rx comprises a leaving group. In some embodiments, Rx comprises a maleimide. In some embodiments, L 3 is as follows: [ka] (In the formula, RX *contains a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached to a residue of the antibody or antigen-binding fragment, and R * Above [ka] represents the point of attachment to a residue of the antibody or antigen-binding fragment thereof, and n=0 to 9) is represented.

[0222] In some aspects, the present disclosure provides: [ka] and a salt of any one thereof.

[0223] In some aspects, the present disclosure provides: [ka] [ka] and any one of the salts thereof (In the formula, RX * comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached to a residue of an antibody or antigen-binding fragment thereof, and R * Above [ka] represents the point of attachment to a residue of the antibody or antigen-binding fragment thereof) The present invention provides a compound or salt selected from:

[0224] In some embodiments, RX * contains a succinamide moiety and is attached to a cysteine ​​residue of the antibody or antigen-binding fragment thereof. * contains a hydrolyzed succinamide moiety and is attached to a cysteine ​​residue of the antibody or antigen-binding fragment thereof.

[0225] In some aspects, the present disclosure provides a compound of the formula: [ka] wherein the antibody is an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein, D is a compound or salt of Category A disclosed herein, and L 3 is the linker part) The conjugate is represented by:

[0226] In some aspects, the present disclosure provides a compound of the formula: [ka] (Wherein, the antibody is an anti-Nectin-4 antibody or an antigen-binding fragment thereof disclosed herein, and DL 3 is a Category A compound or salt disclosed herein) The conjugate is represented by:

[0227] In some aspects, the present disclosure provides a pharmaceutical composition comprising a conjugate disclosed herein and at least one pharmaceutically acceptable excipient.

[0228] In some embodiments, the average DAR of the conjugate is from about 2 to about 8, or from about 1 to about 3, or from about 3 to about 5.

[0229] Examples of TLR8 agonist compounds and their stereoisomers according to Category A are provided in Table 1a. It is understood that as compounds are provided in Table 1a, salts of the compounds are contemplated by Table 1a. [Table 1a-1] [Table 1a-2] [Table 1a-3] [Table 1a-4] [Table 1a-5] [Table 1a-6] [Table 1a-7] [Table 1a-8] [Table 1a-9]

[0230] Category B compounds, TLR7 agonists In some aspects, the present disclosure provides a compound having the structure of formula (IA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 1 , R 2 , R 3 , R 4 , and R 5 is hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN); or R 3 and R11 together, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN; R 6 is halogen, -OR 20 , -N(R 20 )2, -C(O)N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -S(O)R 20 , and -S(O)R 20 ; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN); R 7 , R 8 , R 9 , and R 10 represents, in each occurrence, hydrogen and halogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen; R 11 and R 12 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, and -CN; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from: carbocycle, and 3- to 12-membered heterocycle; or R 11 and R 12 together, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and C optionally substituted with one or more substituents independently selected from —CN3~6 forming a carbocyclic ring; R 13 and R 14 represents, in each occurrence, hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, and -CN;C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6alkynyl; R 15 In each occurrence, represents a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which may contain halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle; R 16 is hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which may contain halogen, -OH, -CN, -NO2, -NH2, =O, =S, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from: carbocycle, and 3- to 12-membered heterocycle; R 20represents, in each occurrence, hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle; X 1 is O, S, or NR 16 and; X 2 is C(O) or S(O)2; n is 1, 2, or 3; x is 1, 2, or 3; w is 0, 1, 2, 3, or 4; z is 0, 1, or 2) The present invention provides a compound represented by the formula:

[0231] In certain embodiments, for compounds of formula (IA), where X 1 is O. In certain embodiments, for compounds of formula (IA), n is 2. In certain embodiments, for compounds of formula (IA), x is 2. In certain embodiments, for compounds of formula (IA), z is 0. In certain embodiments, for compounds of formula (IA), z is 1.

[0232] In certain embodiments, the compound of formula (IA) has formula (IB): [ka] or a pharmaceutically acceptable salt thereof (In the formula, R7’ , R 7’’ , R 8’ , R 8’’ , R 9’ , R 9’’ , R 10’ , and R 10’’ represents, in each occurrence, hydrogen and halogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen is expressed by In certain embodiments, the compound of formula (IA) has the formula (IC): [ka] or a pharmaceutically acceptable salt thereof (In the formula, R 7’ , R 7’’ , R 8’ , R 8’’ , R 9’ , R 9’’ , R 10’ , and R 10’’ represents, in each occurrence, hydrogen and halogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen is expressed by

[0233] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 1 , R 2 , R 3 , R 4 , and R 5 represents hydrogen, as well as halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and C optionally substituted with one or more substituents independently selected from —CN 1~6 alkyl.

[0234] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 1 and R 2 is hydrogen and C 1~6 In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 1 and R 2 are hydrogen atoms.

[0235] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 3 is hydrogen and C optionally substituted with one or more halogens 1~6 alkyl.

[0236] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 3 is hydrogen.

[0237] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 4 is hydrogen and C optionally substituted with one or more halogens 1~6 alkyl.

[0238] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 4 is hydrogen.

[0239] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 5 represents hydrogen, as well as halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and C optionally substituted with one or more substituents independently selected from —CN 1~6 In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 5 is hydrogen.

[0240] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 6 is halogen, -OR 20 , and -N(R 20 )2; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN); R 20 represents, in each occurrence, hydrogen; and C1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0241] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC): R 6 is halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 C optionally substituted with one or more substituents independently selected from 1~6 is alkyl; R 20 represents, in each occurrence, hydrogen; C 1~6 Alkyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0242] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 6 -OR 20 C replaced with 1~6 alkyl, and R 20 is C optionally substituted with hydrogen and one or more substituents independently selected from halogen, —OH, and —NH 1~6 alkyl.

[0243] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 7’ , R 7’’ , R 8’ , R 8’’ , R 9’ , R 9’’ , R 10’ , and R 10’’ is, in each occurrence, C optionally substituted with one or more substituents independently selected from hydrogen and halogen; and halogen. 1~6 alkyl.

[0244] In certain embodiments, for a compound or salt of any one of formula (IB) or (IC), R 7’ and R 8’ and are each hydrogen. In certain embodiments, for a compound or salt of any one of formula (IB) or (IC), R 7’’ and R 8’’ are C 1~6 In certain embodiments, for a compound or salt of any one of formula (IB) or (IC), R 7’’ and R 8’’ are methyl, respectively.

[0245] In certain embodiments, for a compound or salt of any one of formula (IB) or (IC), R 9’ , R 9’’ , R 10’ , and R 10’’ In each occurrence, hydrogen and C 1~6 alkyl.

[0246] In certain embodiments, for a compound or salt of any one of formula (IB) or (IC), R 9’ , R 9’’ , R 10’ , and R 10’’ are hydrogen atoms.

[0247] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 11 and R 12 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 ; and halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , C 3~12 C optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle 1~6 alkyl.

[0248] In certain embodiments, for a compound or salt of any one of formula (IA) or (IC), R 13 and R 14 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20)2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 ; and halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , C 3~12 C optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle 1~6 alkyl.

[0249] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 3 and R 11 taken together form an optionally substituted 5- to 6-membered heterocycle.

[0250] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), R 11 and R 12 are taken together and replaced as necessary by C 3~6 Forms a carbocyclic ring.

[0251] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), X 2 is C(O). In certain embodiments, the compound is [ka] [ka] or a pharmaceutically acceptable salt of any one of them.

[0252] In certain aspects, the present disclosure provides a pharmaceutical composition of a compound or a pharmaceutically acceptable salt of any one of formulas (IA), (IB), or (IC), and a pharmaceutically acceptable excipient.

[0253] In certain embodiments, for a compound or salt of any one of formula (IA), (IB), or (IC), the compound or salt may further comprise a linker, L 3 is further covalently bonded to

[0254] In certain aspects, the present disclosure provides a compound of formula (IIA): [ka] or a pharmaceutically acceptable salt thereof (In the formula: R 2 and R 4 is hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN); R 21 , R 23 , and R 25 is hydrogen; C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN; and L 3 or independently selected from R 23 and R 11 together, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN; 21 , R 23 , and R 25 One of them is L 3 and; R 6 is halogen, -OR 20 , -N(R 20 )2, -C(O)N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -S(O)R 20 , and -S(O)R 20 ; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and —CN); R 7 , R 8 , R 9 , and R 10 represents, in each occurrence, hydrogen and halogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen; R 11 and R 12 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, and -CN; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from: carbocycle, and 3- to 12-membered heterocycle; or R 11 and R 12together, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and C optionally substituted with one or more substituents independently selected from —CN 3~6 forming a carbocyclic ring; R 13 and R 14 represents, in each occurrence, hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 3~12 optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle; C 3~12 Carbocyclic and 3- to 12-membered heterocyclic rings (each of which may contain halogen, -OR 20 , -SR 20 , -C(O)N(R 20)2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl; R 15 In each occurrence, represents a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), -CN, C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which may contain halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle; R 16 is hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12Carbocycles, and 3- to 12-membered heterocycles (each of which may contain halogen, -OH, -CN, -NO2, -NH2, =O, =S, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 optionally substituted with one or more substituents independently selected from: carbocycle, and 3- to 12-membered heterocycle; R 20 represents, in each occurrence, hydrogen; C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle; L 3 is a linker; X 1 is O, S, or NR 16 and; X 2 is C(O) or S(O)2; n is 1, 2, or 3; x is 1, 2, or 3; w is 0, 1, 2, 3, or 4; z is 0, 1, or 2) The present invention provides a compound represented by the formula:

[0255] In certain embodiments, for a compound or salt of formula (IIA), X 1is O. In certain embodiments, for a compound or salt of Formula (IIA), n is 2. In certain embodiments, for a compound or salt of Formula (IIA), x is 2. In certain embodiments, for a compound or salt of Formula (IIA), z is 0. In certain embodiments, for a compound or salt of Formula (IIA), z is 1.

[0256] In certain embodiments, the compound of formula (IIA) is (IIB) or (IIC): [ka] or a pharmaceutically acceptable salt thereof (In the formula, R 7’ , R 7’’ , R 8’ , R 8’’ , R 9’ , R 9’’ , R 10’ , and R 10’’ represents, in each occurrence, hydrogen and halogen; and C 1~6 Alkyl, C 2~6 Alkenyl, and C 2~6 alkynyl, each of which is optionally substituted with one or more substituents independently selected from halogen is expressed by

[0257] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 2 and R 4 represents hydrogen, as well as halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20), and C optionally substituted with one or more substituents independently selected from —CN 1~6 alkyl.

[0258] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 2 and R 4 is hydrogen and C 1~6 In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 2 and R 4 are hydrogen atoms.

[0259] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 23 is hydrogen and C optionally substituted with one or more halogens 1~6 In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC), R 23 is hydrogen.

[0260] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 21 is hydrogen and C optionally substituted with one or more halogens 1~6 In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC), R 21 is hydrogen.

[0261] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 21 L 3 is.

[0262] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 25 represents hydrogen, as well as halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R 20 ), and C optionally substituted with one or more substituents independently selected from —CN 1~6 In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC), R 25 is hydrogen.

[0263] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 25 L 3 is.

[0264] In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC): R 6 is halogen, -OR 20 , and -N(R 20 )2; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl (each of which is a halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , -NO2, =O, =S, =N(R20 ), and —CN); R 20 represents, in each occurrence, hydrogen; and C 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C 1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from:

[0265] In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC): R 6 is halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -S(O)R 20 , -S(O)2R 20 , -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 C optionally substituted with one or more substituents independently selected from 1~6 is alkyl; R 20 represents, in each occurrence, hydrogen, -NH2, -C(O)OCH2C6H5;C 1~6 Alkyl, C 3~12 Carbocycles, and 3- to 12-membered heterocycles (each of which is substituted with halogen, -OH, -CN, -NO2, -NH2, =O, =S, -C(O)OCH2C6H5, -NHC(O)OCH2C6H5, C1~6 Alkyl, -C 1~6 Haloalkyl, -OC 1~6 Alkyl, C 2~6 Alkenyl, C 2~6 Alkynyl, C 3~12 carbocycle, and 3- to 12-membered heterocycle, optionally substituted with one or more substituents independently selected from: In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC): R 6 -OR 20 C replaced with 1~6 is alkyl, R 20 is hydrogen and C 1~6 alkyl, which is optionally substituted with one or more substituents independently selected from halogen, -OH, and -NH2.

[0266] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 7’ , R 7’’ , R 8’ , R 8’’ , R 9’ , R 9’’ , R 10’ , and R 10’’ is, in each occurrence, C optionally substituted with one or more substituents independently selected from hydrogen and halogen; and halogen. 1~6 alkyl.

[0267] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 7’ and R 8’ is hydrogen.

[0268] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 7’’ and R 8’’ is C 1~6It is alkyl.

[0269] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 7’’ and R 8’’ is methyl.

[0270] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 9’ , R 9’’ , R 10’ , and R 10’’ In each occurrence, hydrogen and C 1~6 alkyl.

[0271] In certain embodiments, for a compound or salt of any one of Formula (IIB) or (IIC), R 9’ , R 9’’ , R 10’ , and R 10’’ are hydrogen atoms.

[0272] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 11 and R 12 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , and -OC(O)R 20 ; and halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , C 3~12 C optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle 1~6alkyl.

[0273] In certain embodiments, for a compound or salt of any one of formula (IIA) or (IIC), R 13 and R 14 is hydrogen, halogen, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , and -OC(O)R 20 ; and halogens, -OR 20 , -SR 20 , -C(O)N(R 20 )2, -N(R 20 )2, -C(O)R 20 , -C(O)OR 20 , -OC(O)R 20 , C 3~12 C optionally substituted with one or more substituents independently selected from carbocycle, and 3- to 12-membered heterocycle 1~6 alkyl.

[0274] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 23 and R 11 taken together form an optionally substituted 5- to 6-membered heterocycle.

[0275] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), R 11 and R 12 are taken together and replaced as necessary by C 3~6 Forms a carbocyclic ring.

[0276] In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC), X 2 is C(O).

[0277] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), L 3 is a cleavable linker. In certain embodiments, for a compound or salt of any one of formula (IIA), (IIB), or (IIC), L 3 is cleavable by lysosomal enzymes.

[0278] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), L 3 is the expression: [ka] (In the formula: L 4 represents the C-terminus of the peptide, and L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 30 and wherein R X is a reactive moiety; R 30 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2; and C1-C 10 Alkyl, C2-C 10 Alkenyl, and C2-C 10 alkynyl (each of which, at each occurrence, is independently optionally substituted with one or more substituents selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, and —NO) is expressed by

[0279] In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), R X comprises a leaving group. In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), R X is maleimide or alpha-halocarbonyl. In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), L 3 The peptide contains Val-Cit or Val-Ala.

[0280] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), L 3 is the expression: [ka] (In the formula: RX comprises a reactive moiety; n is 0 to 9) is expressed by

[0281] In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), R X comprises a leaving group. In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), R X is maleimide or alpha-halocarbonyl. In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), or (IIC), L 3 is further covalently attached to an antibody or antigen-binding fragment thereof to form a conjugate.

[0282] In certain embodiments, the present disclosure provides a compound of the formula: [ka] (In the formula: the antibody is an anti-Nectin-4 antibody or an antigen-binding fragment thereof disclosed herein; n is 1 to 20; D is a compound or salt of any one of the compounds of formula (IA), (IB), or (IC) in Category B; L 3 is a linker moiety; or DL 3 is a compound or salt of any one of the compounds of formula (IIA), (IIB), or (IIC) in Category B. The conjugate is represented by:

[0283] In certain embodiments, for conjugates of a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC), n is selected from 1 to 8. In certain embodiments, for conjugates of a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC), n is selected from 2 to 5. In certain embodiments, for conjugates of a compound or salt of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC), n is 2.

[0284] In certain embodiments, for a compound or salt of any one of Formulas (IIA), (IIB), and (IIC), -L 3 is the expression: [ka] (In the formula: L 4 represents the C-terminus of the peptide, and L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 30 and optionally substituted with one or more groups independently selected from: RX * is a bond, succinimide moiety, or hydrolyzed succinimide moiety attached to a residue of an antibody or antigen-binding fragment thereof, and RX * Above [ka] represents the point of attachment to a residue of an antibody or antigen-binding fragment thereof; R 30 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2; and C1-C 10 Alkyl, C2-C 10 Alkenyl, and C2-C 10 alkynyl (each of which, at each occurrence, is independently optionally substituted with one or more substituents selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, and —NO) is expressed by

[0285] In certain embodiments, for a compound or salt of any one of Formula (IIA), (IIB), or (IIC), RX * is a succinamide moiety, a hydrolyzed succinamide moiety, or a mixture thereof, which is attached to a cysteine ​​residue of the antibody.

[0286] In certain embodiments, for compounds of formula (IIA), (IIB), and (IIC), -L 3 is the expression: [ka] (In the formula: RX * is a bond, succinimide moiety, or hydrolyzed succinimide moiety attached to a residue of the antibody, and RX * Above [ka] represents the point of attachment to the antibody residue; n is 0 to 9) is expressed by

[0287] Examples of TLR7 agonist compounds and their stereoisomers according to Category B are provided in Table 3. It is understood that salts of the compounds provided in Table 3 are also contemplated by Table 3. Table 3: Compounds 3.1~3.14 [Table 3-1] [Table 3-2] [Table 3-3]

[0288] In some aspects, the present disclosure provides a compound having the following structure: [ka] or a pharmaceutically acceptable salt thereof wherein Ab comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein, and D is a compound of category B of formula (IID): [ka] (In the formula, R 4 is an alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl group containing 1 to 8 carbons, each J is hydrogen, each U is N, each t is 2, Q is absent, the dashed line represents the point of attachment of the adjuvant to G1, G1 is a bond; the subscript a is an integer from 1 to 40; and the subscript r is an integer from 1 to 10). (a compound or salt of The conjugate is represented by:

[0289] In certain embodiments, D has the following structure: [ka] It has.

[0290] In a further embodiment, the conjugate has the following structure: [ka] It has.

[0291] [ka] wherein D is a compound of category B of formula (IID): [ka] (In the formula, R 4 is an alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, arylalkyl, or heteroarylalkyl group containing 1 to 8 carbons, each J is hydrogen, each U is N, each t is 2, Q is absent, the dashed line represents the point of attachment of the adjuvant to G1, and G1 is a bond; the subscript a is an integer from 1 to 40; and the subscript r is an integer from 1 to 10); or [ka] (a compound or salt of In any of the foregoing embodiments having a conjugate structure of (1) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; or VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; (2) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 33, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 35; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 36, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 37, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; (3) VH comprises CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 39, VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 41; or VL comprises CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 42, VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 43, and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 44; (4) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 45, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 46, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 48, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 49, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 50; (5) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 54, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 55, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 56; (6) VH comprises CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 57, VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 58, and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 59; or VL comprises CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 60, VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 62; (7) VH comprises the amino acid sequence of SEQ ID NO: 10, and VL comprises an amino acid sequence selected from any one of SEQ ID NOs: 12 to 17; or (8) VH comprises the amino acid sequence of SEQ ID NO: 24, and VL comprises the amino acid sequence selected from any one of SEQ ID NOs: 26 to 31.

[0292] In another aspect, the present disclosure provides a compound having the following structure: [ka] or a pharmaceutically acceptable salt thereof where Ab comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein; A is an unmodified amino acid side chain in the antibody or a modified amino acid side chain in the antibody; Z is a linking moiety; and R 1 H and C 1~4 alkyl; or Z, R 1 and the nitrogen atom to which they are attached form a linking moiety comprising a 5- to 8-membered heterocycle; each Y is independently CHR 2 (In the formula, R 2 is selected from H, OH, and NH2; and R 3 is C 1~6 alkyl and 2-6 membered heteroalkyl, each of which is optionally substituted with one or more members selected from the group consisting of halo, hydroxy, amino, oxo (=O), alkylamino, amido, acyl, nitro, cyano, and alkoxy; X is selected from O and CH; the subscript n is an integer from 1 to 12; and the subscript r is an integer from 1 to 10. The conjugate is represented by:

[0293] In certain embodiments, the conjugate has the following structure: [ka] or a pharmaceutically acceptable salt thereof where Ab comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein; A is an unmodified amino acid side chain in the antibody or a modified amino acid side chain in the antibody; Z is a linking moiety; and R 1 H and C 1~4 alkyl; or Z, R 1 and the nitrogen atom to which they are attached form a linking moiety comprising a 5- to 8-membered heterocycle; each Y is independently CHR 2 (In the formula, R 2 is selected from H, OH, and NH; X is selected from O and CH; the subscript n is an integer from 1 to 12; and W is selected from the group consisting of O and CH is expressed by

[0294] In a further embodiment, the conjugate has the following structure: [ka] or a pharmaceutically acceptable salt thereof (wherein Ab comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein; the subscript r is an integer from 1 to 10; A is an unmodified amino acid side chain in the antibody or a modified amino acid side chain in the antibody; Z is a linking moiety; and R 1 H and C 1~4 alkyl; or Z, R 1 and the nitrogen atom to which they are attached form a linking moiety comprising a 5- to 8-membered heterocycle; R 2 is selected from H, OH and NH is expressed by

[0295] In still further embodiments, the conjugate has the following structure: [ka] or a pharmaceutically acceptable salt thereof (wherein Ab comprises an anti-Nectin-4 antibody or antigen-binding fragment thereof disclosed herein; A is an unmodified amino acid side chain in the antibody or a modified amino acid side chain in the antibody; R 2 is selected from H, OH, and NH; and the subscript r is an integer from 1 to 10. is expressed by

[0296] [ka] In any of the foregoing embodiments having a conjugate structure of (1) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; or VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; (2) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 33, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 35; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 36, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 37, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; (3) VH comprises CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 39, VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 41; or VL comprises CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 42, VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 43, and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 44; (4) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 45, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 46, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 48, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 49, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 50; (5) VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; or VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 54, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 55, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 56; (6) VH comprises CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 57, VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 58, and VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 59; or VL comprises CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 60, VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 62; (7) VH comprises the amino acid sequence of SEQ ID NO: 10, and VL comprises an amino acid sequence selected from any one of SEQ ID NOs: 12 to 17; or (8) VH comprises the amino acid sequence of SEQ ID NO: 24, and VL comprises the amino acid sequence selected from any one of SEQ ID NOs: 26 to 31. Category C compounds, TLR8 agonists

[0297] In some embodiments, the myeloid cell agonist is a benzazepine compound (Bza). In some embodiments, the present disclosure provides compounds of Formula I: [ka] (Wherein: A is an anti-Nectin-4 antibody or an antigen-binding fragment thereof, L is a linker; D xis an immunostimulatory compound; n is selected from 1 to 20; and z is selected from 1 to 20. The conjugate is represented by:

[0298] In certain embodiments of the conjugates of the present disclosure, D x is selected from compounds or salts of compounds of the present disclosure, including, but not limited to, Category C (e.g., Formulas (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), and (Ih)).

[0299] In certain embodiments, L is a group of the formula: [ka] (In the formula, L 4 represents the C-terminus of the peptide, L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 32 and optionally substituted with one or more groups independently selected from: RX * comprises a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached to a residue of an antibody or antigen-binding fragment thereof, RX * Above [ka] represents the point of attachment to a residue of an antibody or antigen-binding fragment thereof; R 32 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2; and C 1~10 Alkyl, C 2~10 Alkenyl, C 2~10alkynyl (each of which is optionally substituted with one or more substituents independently selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, —NO) In some embodiments, the peptide of L comprises Val-Cit or Val-Ala.

[0300] In certain embodiments of compounds of category C of the present disclosure, D x is the expression: [ka] (R 1 , R 2 , R 3 , and R 4 H, C1~C 12 Alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 12 Carbocyclyl, C6-C 20 Aryl, C2-C9 heterocyclyl, and C1-C 20 and heteroaryl, wherein alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heterocyclyl, and heteroaryl are independently selected from the group consisting of -(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 12 alkyldiyl)-N(R 5 )2;-(C3~C 12 Carbocyclyl;-(C3~C 12 Carbocyclyl)- * ;-(C3~C 12 Carbocyclyl)-(C1-C 12 Alkyldiyl)-NR 5 - * ;-(C3~C 12 Carbocyclyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C3~C 12 Carbocyclyl)-NR 5 -C(=NR 5 )NR5 - * ;-(C6~C 20 Aryl);-(C6~C 20 Aryl)- * ;-(C6~C 20 Aryldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C6~C 20 Aryldiyl)-(C1-C 12 Alkyldiyl)-NR 5 -C(=NR 5a )N(R 5 )- * ;-(C2~C 20 Heterocyclyl;-(C2-C 20 Heterocyclyl)- * ;-(C2-C9 heterocyclyl)-(C1-C 12 Alkyldiyl)-NR5- * ;-(C2-C9 heterocyclyl)-(C1-C 12 -(C2-C9 heterocyclyl)-NR 5 -C(=NR 5a )NR 5 - * ;-(C1~C 20 Heteroaryl);-(C1-C 20 Heteroaryl)- * ;-(C1~C 20 Heteroaryl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 20 Heteroaryl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C1~C 20 Heteroaryl)-NR 5 -C(=NR 5a )N(R5 )- * ;-C(=O)- * ;-C(=O)-(C2~C 20 Heterocyclyldiyl)- * ;-C(=O)N(R 5 )2;-C(=O)N(R 5 )- * ;-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)R 5 ;-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)N(R 5 )2;-C(=O)NR 5 -(C1~C 12 alkyldiyl)-N(R 5 )CO2R 5 ;-C(=O)NR 5 -(C1~C 12 alkyldiyl)-N(R 5 )C(=NR5a)N(R 5 )2;-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 C(=NR 5a )R 5 ;-C(=O)NR 5 -(C1-C5 alkyldiyl)-NR 5 (C2-C5 heteroaryl); -C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-N(R 5 )- * ;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)- * ;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C2-C 20Heterocyclyldiyl)-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5* ;-N(R 5 )2;-N(R 5 )- * ;-N(R 5 )C(=O)R 5 ;-N(R 5 )C(=O)- * ;-N(R 5 )C(=O)N(R 5 )2;-N(R 5 )C(=O)N(R 5 )- * ;-N(R 5 )CO2R 5 ;-NR 5 C(=NR 5a )N(R 5 )2;-NR 5 C(=NR 5a )N(R 5 )- * ;-NR 5 C(=NR 5a )R 5 ;-N(R 5 )-(C2-C5 heteroaryl); -O-(C1-C 12 Alkyl); -O-(C1-C 12 alkyldiyl)-N(R 5 )2;-O-(C1~C 12 alkyldiyl)-N(R 5 )- * ;-S(=O)2-(C2~C 20 Heterocyclyldiyl)- * ;-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 Alkyldiyl)-NR 5* ; and -S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12optionally substituted with one or more groups selected from R 2 and R 3 together form a 5- or 6-membered heterocyclyl ring; X 1 , X 2 , X 3 , and X 4 is a bond, C(=O), C(=O)N(R 5 ), O, N(R 5 ), S, S(O)2, and S(O)2N(R 5 ) independently selected from the group consisting of: R 5 H, C6~C 20 Aryl, C6-C 20 Aryldiyl, C1-C 12 Alkyl, and C1-C 12 alkyldiyl; or two R groups together form a 5- or 6-membered heterocyclyl ring; R 5a C6~C 20 Aryl and C1-C 20 selected from the group consisting of heteroaryl; asterisk * indicates the binding site of L, and R 1 , R 2 , R 3 and R 4 one of which is attached to L; L is -C(=O)-(PEG)-;-C(=O)-(PEG)-C(=O)-;-C(=O)-(PEG)-O-;-C(=O)-(PEG)-C(=O)-(PEP)-;-C(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-;-C(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C5 monoheterocyclyldiyl)-; -C(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12alkyldiyl)-(MCgluc)-;-C(=O)-(PEG)-C(=O)-(MCgluc)-;-C(=O)-(PEG)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-;-C(=O)-(PEG)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C5 monoheterocyclyldiyl)-; -C(=O)-(PEG)-N(R 5 )-;-C(=O)-(PEG)-N(R 5 )-(PEG)-C(=O)-(PEP)-;-C(=O)-(PEG)-N+(R 5 )2-(PEG)-C(=O)-(PEP)-;-C(=O)-(PEG)-C(=O)-N(R 5 )CH(AA1)C(=O)-(PEG)-C(=O)-(PEP)-;-C(=O)-(PEG)-C(=O)-N(R 5 )CH(AA1)C(=O)-N(R 5 )-(C1~C 12 Alkyldiyl)-;-C(=O)-(PEG)-SS-(C1-C 12 alkyldiyl)-OC(=O)-;-C(=O)-(PEG)-SS-(C1-C 12 Alkyldiyl)-C(=O)-;-C(=O)-(C1-C 12 alkyldiyl)-C(=O)-(PEP)-;-C(=O)-(C1-C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 Alkyldiyl)-;-C(=O)-(C1-C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )-C(=O);-C(=O)-(C1~C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5)C(=O)-(C2-C5 monoheterocyclyldiyl)-; -C(=O)-CH2CH2OCH2CH2-(C1-C 20 Heteroaryldiyl)-CHO-(PEG)-C(=O)(MCgluc)-;-C(=O)-CHCHOCHCH-(C1-C 20 heteroaryldiyl)-CHO-(PEG)-C(=O)(MCgluc)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C5 monoheterocyclyldiyl)-; and -(succinimidyl)-(CH2) m -C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(═O)—(C2-C5 monoheterocyclyldiyl)-; PEG has the formula: -(CH2CH2O) n -(CH2) m m is an integer from 1 to 5 and n is an integer from 2 to 50; PEP has the formula: [ka] wherein AA1 and AA2 are independently selected from amino acid side chains; or AA1 or AA2 and the adjacent nitrogen atom form a five-membered ring praline amino acid, with the wavy line indicating the point of attachment; R 6 C6~C 20 Aryldiyl and C1-C 20 selected from the group consisting of heteroaryldiyl; is substituted with -CH2O-C(=O)-, and [ka] substituted as needed with ; MCgluc is a compound of the group: [ka] wherein q is 1 to 8 and AA is an amino acid side chain; Alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl are independently selected from F, Cl, Br, I, -CN, -CH3, -CH2CH3, -CH=CH2, -C=CH, -C=CCH3, -CH2CH2CH3, -CH (CH3)2, -CH2CH(CH3)2, -CH2OH, -CH2OCH3, -CH2CH2OH, -C(CH3)2OH, -CH(OH)CH(CH3)2, -C(CH3)2CH2OH, -CH2CH 2SO2CH3, -CH2OP(O)(OH)2, -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CHF2, -CH(CH3)CN, -C(CH3)2CN, -CH2CN, -CH2NH2 , -CH2NHSO2CH3, -CH2NHCH3, -CH2N(CH3)2, -CO2H, -COCH3, -CO2CH3, -CO2C(CH3)3, -COCH(OH)CH3, -CONH2, -CO NHCH3, -CON(CH3)2, -C(CH3)2CONH2, -NH2, -NHCH3, -N(CH3)2, -NHCOCH3, -N(CH3)COCH3, -NHS(O)2CH3, -N(CH3) C(CH3)2CONH2, -N(CH3)CH2CH2S(O)2CH3, -NHC(=NH)H, -NHC(=NH)CH3, -NHC(=NH)NH2, -NHC(=O)NH2, -NO2, =O, - OH, -OCH3, -OCH2CH3, -OCH2CH2OCH3, -OCH2CH2OH, -OCH2CH2N(CH3)2, -O(CH2CH2O)n-(CH2)mCO2H, -O(CH2CH2O) n optionally substituted with one or more groups independently selected from -H, -OP(O)(OH), -S(O)N(CH), -SCH, -S(O)CH, and -S(O)H It contains an aminobenzazepine moiety having the formula:

[0301] In certain embodiments, PEP is selected from the group: [ka] [ka] where n is one or more and AA is an amino acid side chain. In certain embodiments of Formula I, each of AAl and AA2 is independently selected from the side chains of a naturally occurring amino acid. In certain embodiments of Formula I, each of AAl and AA2 is independently selected from H, -CH3, -CH(CH3)2, -CH2(CH6H5), -CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, -CH2CH(CH3)2, -CH2SO3H, and -CH2CH2CH2NHC(O)NH2. In certain embodiments of Formula I, AAl is -CH(CH3)2 and AA2 is -CH2CH2CH2NHC(O)NH2. In certain embodiments of Formula I, each of AAl and AA2 is independently selected from GlcNAc, aspartic acid, -CH2SO3H, and -CH2OPO3H.

[0302] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, LD x are represented by formulas Ia to Id: [ka] [ka] In certain embodiments of the disclosed formulas, including Formula I in Category C, L is -C(=O)-(PEG)- or -C(=O)-(PEG)-C(=O)-. In certain embodiments of the disclosed formulas, including Formula I in Category C, L x is the formula Ie and the formula If: [ka] (Wherein, R in formula If 5ais phenyl optionally substituted with one or more groups selected from F, Cl, Br, I, —CN, and —NO2 In certain embodiments of formulas of the present disclosure, including Formula I in Category C, LD x are represented by formulae Ig and Ih: [ka] is selected from.

[0303] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, L is -C(=O)-(PEG)-C(=O)-(PEP)-.

[0304] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 2 and R 3 are each C1 to C5 alkyl.

[0305] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 2 and R 3 are -CH2CH2CH3, respectively.

[0306] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, X 2 and X 3 are bonds, and R 2 or R 3 is -O-(C1~C 12 alkyl).

[0307] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, X 2 and X 3 are bonds, and R 2 or R 3 is -OCH2CH3.

[0308] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, one of R1 and R4 is -(C1-C 12 alkyldiyl)-N(R5 )- * ;-(C1~C 12 alkyldiyl)-N(R 5 )C(=NR 5 )N(R 5 )- * ;-(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)- * ;-(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-C(=O)- * ;-(C6~C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-C(=O)-(C2-C 20 Heterocyclyldiyl)- * ;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)- * and -C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C2-C 20 Heterocyclyldiyl)-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 - * is selected from.

[0309] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 2 and R 3 One of them is -(C1~C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 12 Alkyldiyl)-O-(C1-C 12alkyldiyl)-N(R 5 )- * ;-(C1~C 12 alkyldiyl)-N(R 5 )C(=NR 5 )-N(R 5 )- * ;-(C1~C 12 Alkyldiyl)-(C6-C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 12 Alkyldiyl)-(C6-C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )-C(=NR 5 )N(R 5 )- * ;-(C2-C6 alkynyldiyl)-N(R 5 )- * and -(C2-C6 alkynyldiyl)-N(R 5 )C(=NR 5 )N(R 5 )- * Selected from X 2 and X 3 is a bond, and the asterisk * indicates the binding site of L.

[0310] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 1 and R 4 One of them is -(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2, and -(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-OH.

[0311] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, C6-C 20 Aryldiyl is phenyldiyl, C2-C 20 Heterocyclyldiyl is azetidinediyl.

[0312] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 1 and R 4 One of them is the formula: [ka] is selected from.

[0313] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, R 1 and R 4 One of them is -C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C2-C 20 Heterocyclyldiyl)-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 -L.

[0314] In certain embodiments of formulas of the present disclosure, including Formula I in Category C, C1-C 20 Heteroaryldiyl is pyridinediyl, C2-C 20 Heterocyclyldiyl is piperidiyl.

[0315] In some aspects, the present disclosure provides a compound of formula (II): [ka] (In the formula, Z is H, -O(C1-C8 alkyl), and N(X 2 R 2 )(X 3 R 3 ) are selected from; R 1 , R 2 , R 3 , and R4 H, C1~C 12 Alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C 12 Carbocyclyl, C6-C 20 Aryl, C2-C9 heterocyclyl, and C1-C 20 and heteroaryl, wherein alkyl, alkenyl, alkynyl, carbocyclyl, aryl, heterocyclyl, and heteroaryl are independently selected from the group consisting of -(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 12 alkyldiyl)-N(R 5 )2;-(C3~C 12 Carbocyclyl;-(C3~C 12 Carbocyclyl)- * ;-(C3~C 12 Carbocyclyl)-(C1-C 12 Alkyldiyl)-NR 5 - * ;-(C3~C 12 Carbocyclyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C3~C 12 Carbocyclyl)-NR 5 -C(=NR 5 )NR 5 - * ;-(C6~C 20 Aryl);-(C6~C 20 Aryl)- * ;-(C6~C 20 Aryldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C6~C 20 Aryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C6~C 20 Aryldiyl)-(C1-C 12 Alkyldiyl)-NR 5-C(=NR 5a )N(R 5 )- * ;-(C2~C 20 Heterocyclyl;-(C2-C 20 Heterocyclyl)- * ;-(C2-C9 heterocyclyl)-(C1-C 12 Alkyldiyl)-NR 5 - * ;-(C2-C9 heterocyclyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C2-C9 heterocyclyl)-NR 5 -C(=NR 5a )NR 5 - * ;-(C1~C 20 Heteroaryl);-(C1-C 20 Heteroaryl)- * ;-(C1~C 20 Heteroaryl)-(C1-C 12 alkyldiyl)-N(R 5 )- * ;-(C1~C 20 Heteroaryl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-(C1~C 20 Heteroaryl)-NR 5 -C(=NR 5a )N(R 5 )- * ;-C(=O)- * ;-C(=O)-(C2~C 20 Heterocyclyldiyl)- * ;-C(=O)N(R 5 )2;-C(=O)N(R 5 )- * ;-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)R 5 ;-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)N(R 5 )2;-C(=O)NR 5-(C1~C 12 alkyldiyl)-N(R 5 )CO2R 5 ;-C(=O)NR 5 -(C1~C 12 alkyldiyl)-N(R 5 )C(=NR 5a )N(R 5 )2;-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 C(=NR 5a a)R 5 ;-C(=O)NR 5 -(C1-C8 alkyldiyl)-NR 5 (C2-C8 heteroaryl); -C(=O)NR 5 -(C1~C 20 heteroaryldiyl)-N(R 5 )- * ;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)- * ;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C2-C 20 Heterocyclyldiyl)-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 - * ;-N(R 5 )2;-N(R 5 )- * ;-N(R 5 )C(=O)R 5 ;-N(R 5 )C(=O)- * ;-N(R 5 )C(=O)N(R 5 )2;-N(R 5 )C(=O)N(R 5 )- * ;-N(R 5 )CO2R 5;-NR 5 C(=NR 5a )N(R 5 )2;-NR 5 C(=NR 5a )N(R 5 )- * ;-NR 5 C(=NR 5a )R 5 ;-N(R 5 )-(C2-C8 heteroaryl); -O-(C1-C 12 Alkyl); -O-(C1-C 12 alkyldiyl)-N(R 5 )2;-O-(C1~C 12 alkyldiyl)-N(R 5 )- * ;-S(=O)2-(C2~C 20 Heterocyclyldiyl)- * ;-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2;-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 Alkyldiyl)-NR 5 - * ; and -S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 or R 2 and R 3 together form a 5- or 6-membered heterocyclyl ring; X 1 , X 2 , X 3 , and X 4 is a bond, C(=O), C(=O)N(R 5 ), O, N(R 5 ), S, S(O)2, and S(O)2N(R 5 ) independently selected from the group consisting of: R 5 H, C6~C 20 Aryl, C6-C 20Aryldiyl, C1-C 12 Alkyl, and C1-C 12 alkyldiyl, or two R 5 the groups together form a 5- or 6-membered heterocyclyl ring; R 5a C6~C 20 Aryl and C1-C 20 selected from the group consisting of heteroaryl; asterisk * indicates the binding site of L, and R 1 , R 2 , R 3 and R 4 one of which is attached to L; L is QC(=O)-(PEG)-;QC(=O)-(PEG)-C(=O)-;QC(=O)-(PEG)-O-;QC(=O)-(PEG)-C(=O)-(PEP)-;QC(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12 Alkyldiyl)-;QC(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C8 monoheterocyclyldiyl)-;QC(=O)-(PEG)-C(=O)N(R 5 )-(C1~C 12 QC(=O)-(PEG)-C(=O)-(MCgluc)-; QC(=O)-(PEG)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 Alkyldiyl)-;QC(=O)-(PEG)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C8 monoheterocyclyldiyl)-;QC(=O)-(PEG)-N(R 5 )-;QC(=O)-(PEG)-N(R 5 )-(PEG)-C(=O)-(PEP)-;QC(=O)-(PEG)-N + (R5 )2-(PEG)-C(=O)-(PEP)-;QC(=O)-(PEG)-C(=O)-N(R 5 )CH(AA1)C(=O)-(PEG)-C(=O)-(PEP)-;QC(=O)-(PEG)-C(=O)-N(R 5 )CH(AA1)C(=O)-N(R 5 )-(C1~C 12 Alkyldiyl)-;QC(=O)-(PEG)-SS-(C1-C 12 Alkyldiyl)-OC(=O)-;QC(=O)-(PEG)-SS-(C1-C 12 Alkyldiyl)-C(=O)-;QC(=O)-(C1-C 12 Alkyldiyl)-C(=O)-(PEP)-;QC(=O)-(C1-C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 Alkyldiyl)-;QC(=O)-(C1-C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )-C(=O);QC(=O)-(C1~C 12 alkyldiyl)-C(=O)-(PEP)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)(C2-C5 monoheterocyclyldiyl)-;QC(=O)-CH2CH2OCH2CH2-(C1-C 20 Heteroaryldiyl)-CHO-(PEG)-C(=O)(MCgluc)-;QC(=O)-CHCHOCHCH-(C1-C 20 heteroaryldiyl)-CHO-(PEG)-C(=O)(MCgluc)-N(R 5 )-(C1~C 12 alkyldiyl)-N(R 5 )C(=O)-(C2-C5 monoheterocyclyldiyl)-; and Q-(CH2) m -C(=O)-(PEP)-N(R 5 )-(C1~C12 alkyldiyl)-N(R 5 )C(═O)—(C2-C5 monoheterocyclyldiyl)-; PEG has the formula: -(CH2CH2O) n -(CH2) m m is an integer from 1 to 5, n is an integer from 2 to 50; PEP has the formula: [ka] wherein AA1 and AA2 are independently selected from amino acid side chains; or AA1 or AA2 and the adjacent nitrogen atom form a five-membered ring praline amino acid, with the wavy line indicating the point of attachment; R 6 C6~C 20 Aryldiyl and C1-C 20 heteroaryldiyl, substituted with -CH2O-C(=O)-; and [ka] substituted as needed with ; MCgluc is a compound of the group: [ka] wherein q is 1 to 8 and AA is an amino acid side chain; Q is N-hydroxysuccinimidyl, N-hydroxysulfosuccinimidyl, maleimide, and F, Cl, NO2, and SO3 - phenoxy substituted with one or more groups independently selected from: Alkyl, alkyldiyl, alkenyl, alkenyldiyl, alkynyl, alkynyldiyl, aryl, aryldiyl, carbocyclyl, carbocyclyldiyl, heterocyclyl, heterocyclyldiyl, heteroaryl, and heteroaryldiyl are defined as F, Cl, Br, I, -CN, -CH3, -CH2CH3, -CH=CH2, -C=CH, -C=CCH3, -CH2CH2CH3, - CH(CH3)2, -CH2CH(CH3)2, -CH2OH, -CH2OCH3, -CH2CH2OH, -C(CH3)2OH, -CH(OH)CH(CH3)2, -C(CH3)2CH2OH, -CH2CH2SO2CH3, -CH2OP(O)(OH)2, -CH2F, -CHF2, -CF3, -CH2CF3, -CH2CHF2, -CH(CH3)CN, -C(CH3)2CN, -CH2 CN, -CH2NH2, -CH2NHSO2CH3, -CH2NHCH3, -CH2N(CH3)2, -CO2H, -COCH3, -CO2CH3, -CO2C(CH3)3, -COCH(OH)C H3, -CONH2, -CONHCH3, -CON(CH3)2, -C(CH3)2CONH2, -NH2, -NHCH3, -N(CH3)2, -NHCOCH3, -N(CH3)COCH3, -N HS(O)2CH3, -N(CH3)C(CH3)2CONH2, -N(CH3)CH2CH2S(O)2CH3, -NHC(=NH)H, -NHC(=NH)CH3, -NHC(=NH)NH2, -NHC(=O)NH2, -NO2, =O, -OH, -OCH3, -OCH2CH3, -OCH2CH2OCH3, -OCH2CH2OH, -OCH2CH2N(CH3)2, -O(CH2CH2O) n -(CH2) m CO2H, -O(CH2CH2O) n optionally substituted with one or more groups independently selected from -H, -OP(O)(OH), -S(O)N(CH), -SCH, -S(O)CH, and -S(O)H The present invention provides a benzazepine-containing conjugate according to the present invention.

[0316] In certain embodiments of formulas of the present disclosure, including formula (II) in category C, PEP is selected from the group: [ka] [ka] where n is one or more and AA is an amino acid side chain. is selected from.

[0317] In certain embodiments of formulas of the present disclosure, including formula (II) in category C, each AA1 and AA2 is independently selected from the side chains of naturally occurring amino acids.

[0318] In certain embodiments of the formulas of the present disclosure, including formula (II) in category C, AA1 and AA2 are independently selected from H, -CH3, -CH(CH3)2, -CH2(C6H5), -CH2CH2CH2CH2NH2, -CH2CH2CH2NHC(NH)NH2, -CH2CH(CH3)2, -CH2SO3H, and -CH2CH2CH2NHC(O)NH2.

[0319] In certain embodiments of formulas of the present disclosure, including formula (II) in category C, each AA1 is -CH(CH3)2 and AA2 is -CH2CH2CH2NHC(O)NH2.

[0320] In certain embodiments of formulas of the present disclosure, including formula (II) in category C, each AA1 and AA2 is independently selected from GlcNAc, aspartic acid, —CH2SO3H, and —CH2OPO3H.

[0321] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, the aminobenzazepine-linker compound of formula (II) is represented by Formulas IIa-IId: [ka] [ka] is selected from.

[0322] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, the aminobenzazepine-linker compounds of formula (II) are represented by Formulae IIe and IIf: [ka] (Wherein R of formula IIf 5a is phenyl optionally substituted with one or more groups selected from F, Cl, Br, I, —CN, and —NO2 is selected from.

[0323] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, L is QC(=O)-(PEG)- or QC(=O)-(PEG)-C(=O)-.

[0324] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, the aminobenzazepine-linker compounds of Formula II are represented by Formulas IIg and IIh: [ka] is selected from.

[0325] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, L is -C(=O)-(PEG)-C(=O)-(PEP)-.

[0326] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, R 2 and R 3 are each C1 to C8 alkyl.

[0327] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, R 2 and R 3 are -CH2CH2CH3, respectively.

[0328] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, X 2 and X 3are bonds, and R 2 and R 3 is -O-(C1~C 12 alkyl).

[0329] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, X 2 and X 3 are bonds, and R 2 and R 3 is -OCH2CH3.

[0330] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, R 1 and R 4 One of them is -(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-N(R 5 )2, and -(C6~C 20 Aryldiyl)-S(=O)2-(C2~C 20 Heterocyclyldiyl)-(C1-C 12 alkyldiyl)-OH.

[0331] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, C6-C 20 Aryldiyl is phenyldiyl, C2-C 20 Heterocyclyldiyl is azetidinediyl.

[0332] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, the aminobenzazepine-linker compound of Formula II has the formula: [ka] [ka] is selected from.

[0333] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, R 1 and R 4 One of them is -C(=O)NR 5 -(C1~C 20 Heteroaryldiyl)-(C2-C 20 Heterocyclyldiyl)-C(=O)NR 5 -(C1~C 12 Alkyldiyl)-NR 5 -L.

[0334] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, C1-C 20 Heteroaryldiyl is pyridinediyl, C2-C 20 Heterocyclyldiyl is piperidiyl.

[0335] In certain embodiments of formulas of the present disclosure, including Formula II in Category C, Q is [ka] is selected from.

[0336] In some aspects, the present disclosure provides a compound of formula III: [ka] a pharmaceutically acceptable salt thereof, or a quaternary ammonium salt thereof (In the formula, R 1 , R 2 , R 3 , and R 4 are independently Y or Z, where R 1 , R 2 , R 3 , and R 4 One of them is the formula: [ka] Y having Each Z is independently hydrogen or a group represented by the formula: [ka] Selected from U is optionally present and is CH, C(=O), CHC(=O), or C(=O)CH; A is optionally present, NR 10 , or the expression: [ka] is selected from R 10 and W are independently hydrogen, Ar 1 , or the expression: [ka] and V is optionally present and has the formula: [ka] It is of J 1 and J. 2 are independently CH or N, m 1 , m 2 , and m 3 are independently integers from 0 to 25, provided that m 1 , m 2 , and m 3 Unless at least one of the is a non-zero integer, n 1 , n 2 , n 3 , n 4 , n 5 , and n 6 are independently an integer from 0 to 10, t1 and t2 are independently an integer of 1 to 3; G1, G2, G3, and G4 are independently CH2, C(=O), CH2C(=O), C(=O)CH2, or a bond; X 1 , X 2 , X 3 , and X 4are optionally present and independently represent O, NR 7 , CHR 7 , SO, S, or one or two cycloalkyldiyl, heterocycloalkyldiyl, aryldiyl, or heteroaryldiyl groups, and when more than one cycloalkyldiyl, heterocycloalkyldiyl, aryldiyl, or heteroaryldiyl group is present, the two or more cycloalkyldiyl, heterocycloalkyldiyl, aryldiyl, or heteroaryldiyl groups are linked or fused together, wherein the linked cycloalkyldiyl, heterocycloalkyldiyl, aryldiyl, or heteroaryldiyl groups are linked through a bond or -CO-; R 9 is hydrogen, C1-C4 alkyl, or a group of the formula: [ka] is selected from R 8 are independently hydrogen or C1-C4 alkyl, Ar 1 and Ar 2 are independently an aryl or heteroaryl group optionally substituted with one or more halogen (e.g., fluorine, chlorine, bromine, or iodine), nitrile, hydroxyl, C1-C4 alkyl group, or combinations thereof; L M is a linking moiety comprising a functional group selected from amide, amine, ester, carbamate, urea, thioether, thiocarbamate, thiocarbonate, and thiourea; r is an integer from 1 to 50, "Ms" is a polymeric support; Each wavy line (~) represents a point of attachment. The present invention provides a benzazepine-containing conjugate according to the present invention.

[0337] Non-limiting examples of Category C TLR8 agonist compounds are provided in Table 1b. [Table 1b-1] [Table 1b-2]

[0338] Linker

[0339] The conjugate comprises a linker that connects the anti-Nectin-4 antibody or its antigen-binding fragment to at least one immunostimulatory compound, such as a myeloid cell agonist. The linker can be, for example, a cleavable linker or a non-cleavable linker. The conjugate can comprise multiple linkers. The linkers in the conjugate can be the same or different linkers.

[0340] As will be understood by those skilled in the art, a linker connects an immunostimulatory compound, such as a myeloid cell agonist, to an antibody or antigen-binding fragment thereof by forming a covalent linkage to the compound at one site and to the antibody or antigen-binding fragment thereof at another site. The covalent linkage can be formed by reaction between functional groups on the linker and functional groups on the immunostimulatory compound and the antibody or antigen-binding fragment thereof. As used herein, the term "linker" can include (i) an unlinked form of the linker, which can include a functional group capable of covalently linking the linker to the immunostimulatory compound and a functional group capable of covalently linking the linker to the antibody or antigen-binding fragment thereof; (ii) a partially linked form of the linker, which can include a functional group capable of covalently linking the linker to the antibody or antigen-binding fragment thereof and can be covalently linked to the immunostimulatory compound, or vice versa; and (iii) a fully linked form of the linker, which can be covalently linked to both the immunostimulatory compound and the antibody or antigen-binding fragment thereof. In some specific embodiments, the functional groups on the linker and the covalent linkage formed between the linker and the antibody or antigen-binding fragment thereof may be specifically designated as Rx and Rx', respectively.

[0341] Linkers can be short or long, cleavable or non-cleavable. Linkers can contain segments with different characteristics, such as flexible or rigid segments, hydrophilic segments, and / or hydrophobic segments. Linkers can be chemically stable to the extracellular environment, for example, chemically stable in the bloodstream, and / or can contain linkages that are not stable. Linkers can contain linkages designed to be specifically or nonspecifically cleaved and / or degraded or otherwise degraded inside cells. Cleavable linkers can be sensitive to enzymes at specific sites, such as lysosomes or the extracellular space adjacent to cancer cells.

[0342] Cleavable linkers may include valine-citrulline peptides, valine-alanine peptides, phenylalanine-lysine peptides, or other peptides, such as peptides that form protease recognition and cleavage sites. Such peptide-containing linkers may contain a pentafluorophenyl group. Peptide-containing linkers may contain a succinimide group or a maleimide group. Peptide-containing linkers may contain a para-aminobenzoic acid (PABA) group. Peptide-containing linkers may contain an aminobenzyloxycarbonyl (PABC) group. Peptide-containing linkers may contain a PABA or PABC group and a pentafluorophenyl group. Peptide-containing linkers may contain a PABA or PABC group and a succinimide group. Peptide-containing linkers may contain a PABA or PABC group and a maleimide group.

[0343] The non-cleavable linker is generally insensitive to proteases and intracellular processes. The non-cleavable linker may comprise a maleimide group. The non-cleavable linker may comprise a succinimide group. The non-cleavable linker may be a maleimide-alkyl-C(O)-linker. The non-cleavable linker may be a maleimidocaproyl linker. The maleimidocaproyl linker may be N-maleimidomethylcyclohexane-1-carboxylate. The maleimidocaproyl linker may comprise a succinimide group. The maleimidocaproyl linker may comprise a pentafluorophenyl group.

[0344] The linker may be a combination of a maleimide caproyl group and one or more polyethylene glycol molecules. The linker may be a maleimide-PEG4 linker. The linker may be a combination of a maleimide caproyl linker containing a succinimide group and one or more polyethylene glycol molecules. The linker may be a combination of a maleimide caproyl linker containing a pentafluorophenyl group and one or more polyethylene glycol molecules. The linker may contain a maleimide linked to a polyethylene glycol molecule, where the polyethylene glycol may allow for greater linker flexibility or allow for longer linkers to be used.

[0345] The linker may be a (maleimidocaproyl)-(valine-alanine)-(para-aminobenzyloxycarbonyl) linker. The linker may be a (maleimidocaproyl)-(valine-citrulline)-(para-aminobenzyloxycarbonyl) linker. The linker may be a (maleimidocaproyl)-(phenylalanine-lysine)-(para-aminobenzyloxycarbonyl) linker.

[0346] The linker may also contain alkylene, alkenylene, alkynylene, polyether, polyester, polyamide, polyamino acid, peptide, polypeptide, cleavable peptide, and / or aminobenzyl-carbamate segments. The linker may contain a maleimide at one end and an N-hydroxysuccinimidyl ester at the other end. The linker may contain a lysine with an acetylated N-terminal amine and a valine-citrulline, valine-alanine, or phenylalanine-lysine cleavage site. The linker may be a link created by microbial transglutaminase, where the link is created between an amine-containing moiety and a moiety engineered to contain glutamine as a result of the enzyme catalyzing bond formation between the acyl group of the glutamine side chain and the primary amine of the lysine chain. The linker may contain a reactive primary amine. The linker may be a Sortase A linker. A Sortase A linker can be generated by the Sortase A enzyme, fusing the LXPTG recognition motif (SEQ ID NO: 32) to an N-terminal GGG motif to regenerate a native amide bond. Thus, the generated linker can link a moiety bound to the LXPTG recognition motif (SEQ ID NO: 32) with a moiety bound to the N-terminal GGG motif. The linker can be a link generated between a non-natural amino acid on one moiety that reacts with an oxime bond formed by modifying a ketone group on another moiety with an alkoxyamine. The moiety can be part of a conjugate. The moiety can be part of an antibody, e.g., an antibody. The moiety can be part of an immunostimulatory compound, e.g., a myeloid cell agonist. The moiety can be part of a binding domain. The linker can be unsubstituted or substituted, e.g., with a substituent. Substituents may include, for example, hydroxyl groups, amino groups, nitro groups, cyano groups, azide groups, carboxyl groups, carboxaldehyde groups, imine groups, alkyl groups, alkenyl groups, alkynyl groups, alkoxy groups, acyl groups, acyloxy groups, amide groups, and ester groups.

[0347] A linker may be multivalent, such that it covalently links two or more immunostimulatory compounds to a single site on an antibody or antigen-binding fragment thereof, or it may be monovalent, such that it covalently links a single immunostimulatory compound to a single site on an antibody or antigen-binding fragment thereof.

[0348] Exemplary polyvalent linkers have been described that can be used to attach many immunostimulatory compounds to the antibody or antigen-binding fragment thereof of a conjugate. For example, Fleximer® linker technology holds promise for enabling high DAR conjugates with favorable physicochemical properties. As shown below, Fleximer® linker technology is based on the incorporation of molecules into a soluble polyacetal backbone via an array of ester bonds. This methodology affords highly loaded conjugates (up to 20 DAR) while maintaining favorable physicochemical properties. This methodology can be utilized with the immunostimulatory compounds shown in the following scheme (where 'drug' refers to the immunostimulatory compound): [ka]

[0349] To utilize the Fleximer® linker technology depicted in the scheme above, an aliphatic alcohol can be present in or introduced into the immunostimulatory compound. The alcohol moiety is then attached to an alanine moiety, which is then synthetically incorporated into the Fleximer® linker. Liposome treatment of the conjugate releases the parent alcohol-containing drug in vitro.

[0350] By way of example and not limitation, some cleavable and non-cleavable linkers that may be included in the conjugates described herein are listed below.

[0351] Cleavable linkers can be cleavable in vitro and in vivo.Cleavable linkers can include chemically or enzymatically unstable or degradable linkages.Cleavable linkers can depend on intracellular processes to release immunostimulatory compounds, such as reduction in the cytoplasm, exposure to acidic conditions in lysosomes, or cleavage by specific proteases or other enzymes within cells.Cleavable linkers can incorporate one or more chemical bonds that are chemically or enzymatically cleavable, while the remainder of the linker can be non-cleavable.

[0352] The linker can contain a chemically unstable group such as a hydrazone group and / or a disulfide group. Linkers containing chemically unstable groups can take advantage of the different properties between plasma and some cytoplasmic compartments. The intracellular conditions that can promote the release of immunostimulatory compounds for hydrazine-containing linkers can be the acidic environment of endosomes and lysosomes, while disulfide-containing linkers can be reduced in the cytosol, which can contain high thiol concentrations, such as glutathione. The plasma stability of linkers containing chemically unstable groups can be increased by introducing steric hindrance using substituents near the chemically unstable group.

[0353] Acid-labile groups such as hydrazones can remain intact during systemic circulation in the neutral pH environment of blood (pH 7.3-7.5) and can undergo hydrolysis upon internalization of the conjugate into the slightly acidic endosomal (pH 5.0-6.5) and lysosomal (pH 4.5-5.0) compartments of cells, releasing the immunostimulatory compound. This pH-dependent release mechanism may be associated with nonspecific release of the immunostimulatory compound. To increase the stability of the hydrazone group of the linker, the linker can be altered by chemical modification, e.g., substitution, allowing for tuning to minimize loss during circulation and achieve more efficient release in the lysosome.

[0354] The hydrazone-containing linker can contain additional cleavage sites, such as additional acid-labile and / or enzymatically labile cleavage sites. Exemplary conjugates comprising hydrazone-containing linkers have, for example, the following structures: [ka] (where D is an immunostimulatory compound, Ab is an antibody or antigen-binding fragment thereof, and n represents the number of compound-attached linkers (LP) attached to the antibody or antigen-binding fragment thereof, respectively. In certain linkers, such as linker (Ia), the linker can include two cleavable groups: a disulfide moiety and a hydrazone moiety. For such linkers, effective release of the unmodified, free immunostimulatory compound can require acidic pH, or disulfide reduction and acidic pH. Linkers such as (Ib) and (Ic) can be effective with a single hydrazone cleavage site.

[0355] Other acid-labile groups that can be included in the linker include cis-aconityl-containing linkers, whose chemistry can accelerate amide hydrolysis under acidic conditions using a carboxylic acid juxtaposed to the amide bond.

[0356] Cleavable linkers can also contain disulfide groups. Disulfides can be thermodynamically stable at physiological pH and can be designed to release immunostimulatory compounds upon internalization into cells, where the cytosol can provide a significantly more reducing environment than the extracellular environment. Cleavage of disulfide bonds can require the presence of a cytosolic thiol cofactor, such as (reduced) glutathione (GSH), such that disulfide-containing linkers can be reasonably stable in the circulation and selectively release immunostimulatory compounds in the cytosol. The intracellular enzyme protein disulfide isomerase, or similar enzymes capable of cleaving disulfide bonds, can also contribute to preferential cleavage of disulfide bonds inside cells. GSH can be present in cells at concentrations ranging from 0.5 to 10 mM, compared to the significantly lower concentrations of GSH, or cysteine, the most abundant low-molecular-weight thiol in the circulation, at approximately 5 μM. Tumor cells, where irregular blood flow can lead to hypoxic conditions, can result in increased activity of reductases and therefore higher glutathione concentrations. The in vivo stability of disulfide-containing linkers can be enhanced by chemical modifications of the linker, for example, the use of steric hindrance adjacent to the disulfide bond.

[0357] Immunostimulatory conjugates that include disulfide-containing linkers have the following structure: [ka] (wherein D is an immunostimulatory compound, Ab is an antibody or antigen-binding fragment thereof, n represents the number of compounds attached to the linker attached to the antibody or antigen-binding fragment thereof, and R is, for example, independently selected from hydrogen or alkyl at each occurrence. Increasing steric hindrance adjacent to the disulfide bond can increase the stability of the linker. Structures such as (IIa) and (IIc) can exhibit increased in vivo stability when one or more R groups are selected from lower alkyl, such as methyl.

[0358] Another type of linker that can be used is a linker that is specifically cleaved by enzymes.For example, the linker can be cleaved by lysosomal enzymes.Such linkers can be peptide-based or contain peptide regions that can act as substrates for enzymes.Peptide-based linkers can be more stable in plasma and extracellular environments than chemically unstable linkers.

[0359] Lysosomal proteolytic enzymes may have very low activity in blood due to endogenous inhibitors and the unfavorable pH value of blood compared to lysosomes, so peptide bonds may have good serum stability.The release of immunostimulatory compounds from antibodies or their antigen-binding fragments may occur due to the action of lysosomal proteases, such as cathepsin and plasmin.These proteases may be present at elevated levels in certain tumor tissues.The linker may be cleavable by lysosomal enzymes.The lysosomal enzyme may be, for example, cathepsin B, cathepsin S, β-glucuronidase, or β-galactosidase.

[0360] The cleavable peptide can be selected from tetrapeptides such as Gly-Phe-Leu-Gly, Ala-Leu-Ala-Leu, dipeptides such as Val-Cit, Val-Ala, and Phe-Lys, or other peptides. Depending on the composition of the peptide, dipeptides may have lower hydrophobicity compared to longer peptides.

[0361] A variety of dipeptide-based cleavable linkers can be used in the immunostimulatory conjugates described herein.

[0362] The enzymatically cleavable linker can include a self-destructing spacer that spatially separates the immunostimulatory compound from the site of enzymatic cleavage. Direct attachment of the immunostimulatory compound to the peptide linker can result in proteolytic release of the immunostimulatory compound or an amino acid adduct of the immunostimulatory compound, thereby impairing its activity. The use of a self-destructing spacer can allow for the release of a fully active, chemically unmodified immunostimulatory compound upon hydrolysis of the amide bond.

[0363] One self-immolating spacer can be a bifunctional para-aminobenzyl alcohol group (PABA), which can be linked to a peptide through its amino group to form an amide bond, while an amine-containing immunostimulatory compound can be attached to a benzylic hydroxyl group of a linker through a carbamate functionality (yielding p-amidobenzylcarbamate, PABC). The resulting pro-immunostimulatory compound can be activated upon protease-mediated cleavage, resulting in a 1,6-elimination reaction, releasing the unmodified immunostimulatory compound, carbon dioxide, and the remainder of the linker. The following scheme depicts the fragmentation of p-amidobenzylcarbamate and release of the immunostimulatory compound: [ka] (where XD represents the unmodified immunostimulatory compound and the carbonyl group adjacent to "peptide" is part of the peptide.) Heterocyclic variants of this self-immolating group have also been described.

[0364] The enzymatically cleavable linker can be a β-glucuronic acid-based linker. The ready release of the immunostimulatory compound can be achieved by cleavage of the β-glucuronide glycosidic bond by the lysosomal enzyme β-glucuronidase. This enzyme can be abundant in lysosomes and may be overexpressed in some tumor types, but extracellular enzymatic activity may be low. A β-glucuronic acid-based linker can be used to avoid the tendency of the immunostimulatory conjugate to undergo aggregation due to the hydrophilic nature of β-glucuronides. In certain embodiments, a β-glucuronic acid-based linker can link an antibody or antigen-binding fragment thereof to a hydrophobic immunostimulatory compound. The following scheme depicts the release of an immunostimulatory compound (D) from an immunostimulatory conjugate containing the β-glucuronic acid-based linker shown below, where Ab represents an antibody or antigen-binding fragment thereof. [ka]

[0365] Various cleavable β-glucuronic acid-based linkers have been described that are useful for linking drugs such as auristatins, camptothecins, and doxorubicin analogs, CBI minor groove binders, and psymberin to antibodies.These β-glucuronic acid-based linkers can be used in the conjugates described herein.In certain embodiments, the enzymatically cleavable linker is a β-galactoside-based linker.β-galactoside is abundant in lysosomes, but has low enzymatic activity outside cells.

[0366] Additionally, immunostimulatory compounds containing phenolic groups can be covalently attached to linkers through the phenolic oxygen. One such linker relies on the methodology of delivering phenols using a diamino-ethane "space link" in conjunction with conventional "PABO"-based self-immolating groups.

[0367] A cleavable linker can include a non-cleavable moiety or segment, and / or a cleavable segment or moiety can be included in an otherwise non-cleavable linker to make it cleavable. By way of example only, polyethylene glycol (PEG) and related polymers can include cleavable groups in the polymer backbone. For example, polyethylene glycol or a polymer linker can include one or more cleavable groups, such as a disulfide, hydrazone, or dipeptide.

[0368] Other degradable linkages that can be included in the linker include ester linkages formed by the reaction of PEG carboxylic acid or activated PEG carboxylic acid with alcohol groups on the immunostimulatory compound, where such ester groups can be hydrolyzed under physiological conditions to release the immunostimulatory compound.Hydrolytically degradable linkages include, but are not limited to, carbonate linkages; imine linkages obtained from the reaction of amines and aldehydes; phosphate ester linkages formed by the reaction of alcohols with phosphate groups; acetal linkages that are the reaction products of aldehydes and alcohols; orthoester linkages that are the reaction products of formates and alcohols; and oligonucleotide linkages formed by phosphoramidite groups, including, but not limited to, those at the ends of polymers and the 5' hydroxyl groups of oligonucleotides.

[0369] The linker can contain an enzymatically cleavable peptide moiety, for example, the linker can be represented by structural formula (IIIa), (IIIb), (IIIc), or (IIId): [ka] or a pharmaceutically acceptable salt thereof, wherein: "peptide" represents a peptide (designated in the N→C orientation, wherein the peptide comprises an amino and a carboxy "terminus") that is cleavable by a lysosomal enzyme; T represents a polymer comprising one or more ethylene glycol units or alkylene chains, or a combination thereof; R ais selected from hydrogen, alkyl, sulfonate, and methylsulfonate; R y is hydrogen, or C 1~4 Alkyl-(O) r -(C 1~4 alkylene) s -G 1 , or C 1~4 Alkyl-(N)-[(C 1~4 Alkylene)-G 1 ]2;R z is C 1~4 Alkyl-(O) r -(C 1~4 alkylene) s -G 2 and;G 1 is SO3H, CO2H, PEG 4-32, or a sugar moiety; G 2 is SO3H, CO2H, or a PEG 4-32 moiety; r is 0 or 1; s is 0 or 1; p is an integer ranging from 0 to 5; q is 0 or 1; x is 0 or 1; y is 0 or 1; [ka] represents the point of attachment of the linker to the immunostimulatory compound; * represents the point of attachment to the rest of the linker) Includes.

[0370] In certain embodiments, the peptide may be selected from natural amino acids, unnatural amino acids, or combinations thereof, hi certain embodiments, the peptide may be selected from a tripeptide or a dipeptide. In certain embodiments, the dipeptide can comprise an L-amino acid and can be selected from Val-Cit; Cit-Val; Ala-Ala; Ala-Cit; Cit-Ala; Asn-Cit; Cit-Asn; Cit-Cit; Val-Glu; Glu-Val; Ser-Cit; Cit-Ser; Lys-Cit; Cit-Lys; Asp-Cit; Cit-Asp; Ala-Val; Val-Ala; Phe-Lys; Lys-Phe; Val-Lys; Lys-Val; Ala-Lys; Lys-Ala; Phe-Cit; Cit-Phe; Leu-Cit; Cit-Leu; Ile-Cit; Cit-Ile; Phe-Arg; Arg-Phe; Cit-Trp; and Trp-Cit, or a salt thereof.

[0371] Exemplary embodiments of linkers according to structural formula (IIIa) are shown below (as shown, the linker comprises a reactive group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] (In the formula, [ka] indicates the site of attachment of the linker to the immunostimulatory compound).

[0372] Exemplary embodiments of linkers according to structural formula (IIIb), (IIIc), or (IIId) that may be included in the conjugates described herein may include the linkers shown below (as shown, the linker may include a reactive group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] [ka] [ka] [ka] [ka] (In the formula, [ka] indicates the binding site for the immunostimulatory compound).

[0373] The linker can contain an enzymatically cleavable sugar moiety, for example, the linker can be represented by structural formula (IVa), (IVb), (IVc), (IVd), or (IVe): [ka] [ka] or a pharmaceutically acceptable salt thereof (wherein: q is 0 or 1; r is 0 or 1; X 1 is CH2, O, or NH; [ka] represents the point of attachment of the linker to the immunostimulatory compound; * represents the point of attachment to the rest of the linker) Includes.

[0374] Exemplary embodiments of linkers according to structural formula (IVa) that may be included in the immunostimulatory conjugates described herein may include the linker shown below (as shown, the linker includes a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0375] Exemplary embodiments of linkers according to structural formula (IVb) that may be included in the conjugates described herein include the linkers shown below (as shown, the linkers include a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0376] Exemplary embodiments of linkers according to structural formula (IVc) that may be included in the conjugates described herein include the linkers shown below (as shown, the linker includes a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0377] Exemplary embodiments of linkers according to structural formula (IVd) that may be included in the conjugates described herein include the linker shown below (as shown, the linker includes a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0378] Exemplary embodiments of linkers according to structural formula (IVe) that may be included in the conjugates described herein include the linkers shown below (as shown, the linkers include a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0379] Although cleavable linkers can provide certain advantages, the linkers constituting the conjugates described herein do not need to be cleavable. With non-cleavable linkers, the release of the immunostimulatory compound may not depend on the different properties between plasma and some cytoplasmic compartments. The release of the immunostimulatory compound may occur after the internalization of the immunostimulatory conjugate via antigen-mediated endocytosis and delivery to the lysosomal compartment, where the antibody or its antigen-binding fragment may be degraded to the amino acid level through intracellular proteolysis. This process can release an immunostimulatory compound derivative formed by the immunostimulatory compound, the linker, and one or more amino acid residues to which the linker was covalently attached. An immunostimulatory compound derivative derived from an immunostimulatory conjugate having a non-cleavable linker may be more hydrophilic and less membrane permeable, which may result in a lower bystander effect and lower nonspecific toxicity compared to an immunostimulatory conjugate having a cleavable linker. Immunostimulatory conjugates with non-cleavable linkers can have greater stability in circulation than immunostimulatory conjugates with cleavable linkers. The non-cleavable linker can include an alkylene chain or can be polymeric, such as based on a polyalkylene glycol polymer, an amide polymer, or can include an alkylene chain, a polyalkylene glycol, and / or an amide polymer segment. The linker can contain a polyethylene glycol segment having 1 to 6 ethylene glycol units.

[0380] The linker may, for example, have the formula: [ka] or a salt thereof (wherein: R a is selected from hydrogen, alkyl, sulfonate, and methylsulfonate; R x is a reactive moiety containing a functional group capable of covalently linking the linker to an antibody or antigen-binding fragment thereof; [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0381] Exemplary embodiments of linkers according to structural formulas (Va)-(Vf) that may be included in the conjugates described herein include the linkers shown below (as shown, the linker comprises a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof: [ka] represents the point of attachment of the linker to the immunostimulatory compound). [ka] [ka]

[0382] In some embodiments, the linker has formula (V): [ka] (In the formula: L 4 represents the C-terminus of the peptide; L 5 is selected from a bond, alkylene, and heteroalkylene; L 5 is R 32 optionally substituted with one or more groups independently selected from: RX * contains a bond, a succinimide moiety, or a hydrolyzed succinimide moiety attached to a residue of a polypeptide such as an antibody, and RX* Above [ka] represents the point of attachment to a residue of a polypeptide such as an antibody, and [ka] represents a point of attachment to a myeloid cell agonist, such as a TLR8 or TLR7 agonist; R 32 represents, in each occurrence, halogen, -OH, -CN, -O-alkyl, -SH, =O, =S, -NH2, -NO2;C 1~10 Alkyl, C 2~10 Alkenyl, and C 2~10 alkynyl (each of which is optionally substituted with one or more substituents independently selected from halogen, —OH, —CN, —O-alkyl, —SH, ═O, ═S, —NH, and —NO) is expressed by

[0383] The linking groups used to attach the linker to the antibody or antigen-binding fragment thereof can be electrophilic in nature, including, for example, maleimide groups, alkynes, alkynoates, allenes and allenoates, activated disulfides, active esters such as NHS esters and HOBt esters, haloformates, acid halides, alkyl, and benzyl halides such as haloacetamides. Technology is also emerging involving "self-stabilizing" maleimides and "crosslinking disulfides" that can be used in accordance with the present disclosure.

[0384] Maleimide groups are frequently used in the preparation of conjugates due to their specificity for reacting with thiol groups of, for example, cysteine ​​groups of antibodies or antigen-binding fragments thereof in the conjugate. The reaction between a thiol group of an antibody or antigen-binding fragment thereof and a drug having a linker containing a maleimide group can be represented by the following scheme: [ka] Proceed as follows.

[0385] A reverse reaction, resulting in maleimide elimination from the thio-substituted succinimide, can also occur. This reverse reaction is undesirable because the maleimide group can subsequently react with other available thiol groups, such as other proteins in the body that have available cysteines. Therefore, the reverse reaction can undermine the specificity of the conjugate. One way to prevent this is to incorporate a basic group into the linking group shown in the above scheme. While not wishing to be bound by theory, the presence of a basic group may increase the nucleophilicity of nearby water molecules, promoting ring-opening hydrolysis of the succinimide group. The hydrolyzed form of the linking group is resistant to deconjugation in the presence of plasma proteins. So-called "self-stabilizing" linkers provide conjugates with improved stability. A representative schematic is shown below. [ka]

[0386] The hydrolysis reaction outlined above can occur at either carbonyl group of the succinimide group, and therefore two possible isomers can occur, as shown below. [ka]

[0387] The identity of the base and the distance between the base and the maleimide group can be modified to tune the rate of hydrolysis of the thio-substituted succinimide group and optimize targeted delivery of the conjugate, for example, by improving the specificity and stability of the conjugate.

[0388] Bases suitable for inclusion in the linkers described herein, e.g., any linker described herein that has a maleimide group prior to conjugation to an antibody or antigen-binding fragment thereof, can promote hydrolysis of the nearby succinimide group that is formed after conjugation of the antibody or antigen-binding fragment thereof to the linker. Bases include, for example, amines (e.g., -N(R 26 )(R 27 )(wherein, R 26 and R 27 H and C 1~6 alkyl), nitrogen-containing heterocycles (e.g., 3- to 12-membered heterocycles containing one or more nitrogen atoms and optionally one or more double bonds), carbocycles or heterocycles (e.g., optionally containing a heteroatom such as a nitrogen atom and substituted with one or more amine groups) substituted with amidine, guanidine, and one or more amine groups, 26 )(R 27 )(wherein, R 26 and R 27 is H or C 1~6 The basic unit may include, for example, a 3-12 membered aromatic or non-aromatic ring substituted with one or more amines of the type -(CH2) m The maleimide group may be separated from the maleimide group by an alkylene chain of the form -, where m is an integer from 0 to 10. The alkylene chain may be optionally substituted with other functional groups as described herein.

[0389] Linkers described herein having a maleimide group include, but are not limited to, -C(O)R, ═O, -CN, -NO, -CX, -X, -COOR, -CONR, -COR, -COX, -SOR, -SOR, -SONHR, -SONNR, PO, R, -P(O)(CH)NHR, -NO, -NR + , -CR=CR2, and -C≡CR (where each R is H and C 1~6alkyl, and each X is independently selected from F, Br, Cl, and I. The self-stabilizing linker may also include an aryl group, e.g., a phenyl group, or a heteroaryl group, e.g., a pyridine group, optionally substituted with an electron-withdrawing group such as those described herein.

[0390] Examples of self-stabilizing linkers are provided, for example, in U.S. Patent Publication No. US2013 / 0309256, which linkers are incorporated herein by reference. It will be understood that self-stabilizing linkers useful in conjugation with immunostimulatory compounds can equivalently be described as unsubstituted maleimide-containing linkers, thio-substituted succinimide-containing linkers, or hydrolyzed and ring-opened thio-substituted succinimide-containing linkers.

[0391] In certain embodiments, the linker is [ka] The compound comprises a stability linker moiety selected from:

[0392] In the scheme provided above, the structure below can be referred to as (maleimido)-DPR-Val-Cit-PAB (where DPR refers to diaminopropionic acid, Val refers to valine, Cit refers to citrulline, and PAB refers to para-aminobenzylcarbonyl). [ka] represents the point of attachment to the immunostimulatory compound.

[0393] A method for bridging pairs of sulfhydryl groups derived from the reduction of native hinge disulfide bonds has been disclosed and is depicted in the schematic diagram below. The advantage of this methodology is the ability to synthesize homogeneous DAR4 conjugates by complete reduction of IgG (generating four pairs of sulfhydryls from intrachain disulfides) followed by reaction with four equivalents of an alkylating agent. Conjugates containing "bridging disulfides" are also claimed to have improved stability. [ka]

[0394] Similarly, maleimide derivatives have been developed that are capable of cross-linking pairs of sulfhydryl groups, as shown below. [ka]

[0395] The linker may have the following structural formula (VIa), (VIb), or (VIc): [ka] or a salt thereof (wherein: R q is H or -O-(CH2CH2O) 11 -CH3; x is 0 or 1; y is 0 or 1; G 2 is -CH2CH2CH2SO3H or -CH2CH2O-(CH2CH2O) 11 -CH3; R w is -O-CH2CH2SO3H or -NH(CO)-CH2CH2O-(CH2CH2O) 12 -CH3; * represents the point of attachment to the rest of the linker).

[0396] Exemplary embodiments of linkers according to structural formulas (VIa) and (VIb) that may be included in the conjugates described herein may include the linkers shown below (as shown, the linker may include a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0397] Exemplary embodiments of linkers according to structural formula (Vic) that may be included in the immunostimulatory conjugates described herein may include the linkers shown below (as shown, the linker may include a group suitable for covalently linking the linker to an antibody or antigen-binding fragment thereof): [ka] [ka] (In the formula, [ka] represents the point of attachment of the linker to the immunostimulatory compound).

[0398] The linker can be attached to the antibody or its antigen-binding fragment at any suitable position.The factors to be considered when selecting the attachment site include whether the linker is cleavable or non-cleavable, the reactive group of the linker for binding to the antibody or its antigen-binding fragment, the chemical nature of the immunostimulatory compound, and the compatibility between the linker and the reactive site on the antibody or its antigen-binding fragment, and the effect of the attachment site on the functional activity of the Fc domain.The linker can be attached to the end of the amino acid sequence of the antibody or its antigen-binding fragment, or to the side chain of the amino acid of the antibody or its antigen-binding fragment, for example, lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, non-natural amino acid residue, or glutamic acid residue. The linker can be attached to the terminus of the amino acid sequence of the Fc domain or Fc region of the antibody or antigen-binding fragment thereof, or can be attached to the side chain of an amino acid in the Fc domain of the antibody or antigen-binding fragment thereof, for example, the side chain of a lysine, serine, threonine, cysteine, tyrosine, aspartic acid, glutamine, unnatural amino acid residue, or glutamic acid residue.

[0399] In some embodiments, the linker is attached to a hinge cysteine ​​in the Fc domain of the antibody. The linker can be attached to the antibody or antigen-binding fragment thereof at a lysine in the light chain constant domain. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered cysteine ​​in the light chain. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered light chain glutamine. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered non-natural amino acid in the light chain. The linker can be attached to the antibody or antigen-binding fragment thereof at a lysine in the heavy chain constant domain. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered cysteine ​​in the heavy chain. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered heavy chain glutamine. The linker can be attached to the antibody or antigen-binding fragment thereof at an engineered non-natural amino acid in the heavy chain. Amino acids can be engineered into the amino acid sequence of an antibody or antigen-binding fragment thereof that can be connected to a linker in a conjugate, as described herein or known to one of skill in the art. The engineered amino acids can be added to an existing sequence of amino acids. The engineered amino acids can replace one or more existing amino acids in the sequence of amino acids.

[0400] The linker can be attached to the antibody or its antigen-binding fragment via a sulfhydryl group. The linker can be attached to the antibody or its antigen-binding fragment via a primary amine. The linker can be a link created between an unnatural amino acid on the antibody by reacting with an oxime bond formed by modifying a ketone group on the immunostimulatory compound with an alkoxyamine.

[0401] As known to those skilled in the art, the linker selected for a particular conjugate can be influenced by a variety of factors, including, but not limited to, the site of attachment to the antibody or antigen-binding fragment thereof (e.g., lys, cys, or other amino acid residue), the structural constraints of the drug pharmacophore, and the lipophilicity of the drug. The particular linker selected for a conjugate should seek to balance these different factors for a particular antibody / drug combination.

[0402] For example, conjugates have been observed to cause the death of bystander antigen-negative cells present in the vicinity of antigen-positive tumor cells. The mechanism of bystander cell killing by conjugates indicates that metabolic products formed during intracellular processing of the conjugate may play a role. Neutral cytotoxic metabolic products produced by the metabolism of the conjugate in antigen-positive cells appear to play a role in bystander cell killing, while charged metabolic products may be prevented from diffusing across the membrane into or from the medium, and therefore cannot affect bystander killing. In certain embodiments, the linker is selected to weaken the bystander effect caused by the cellular metabolic products of the conjugate. In certain embodiments, the linker is selected to increase the bystander effect.

[0403] The properties of the linker or linker-compound can also affect conjugate aggregation under conditions of use and / or storage. Typically, conjugates reported in the literature contain no more than three to four drug molecules per antibody molecule. Attempts to achieve higher drug-to-antibody ratios ("DAR") often fail due to conjugate aggregation, especially when both the drug and linker are hydrophobic. In many instances, a DAR higher than three to four can be beneficial as a means of increasing efficacy. In cases where the immunostimulatory compound is more hydrophobic in nature, it may be desirable to select a relatively hydrophilic linker as a means of reducing conjugate aggregation, especially when a DAR higher than three to four is desired. Therefore, in certain embodiments, the linker incorporates a chemical moiety that reduces conjugate aggregation during storage and / or use. The linker may incorporate polar or hydrophilic groups, such as charged groups or groups that become charged at physiological pH, to reduce conjugate aggregation. For example, the linker may incorporate a charged group, such as a salt or a group that deprotonates, eg, a carboxylate, or protonates, eg, an amine, at physiological pH.

[0404] In certain embodiments, aggregation of the conjugate during storage or use is less than about 40% as determined by size exclusion chromatography (SEC). In certain embodiments, aggregation of the conjugate during storage or use is less than 35%, such as less than about 30%, for example less than about 25%, for example less than about 20%, for example less than about 15%, for example less than about 10%, for example less than about 5%, for example less than about 4%, or even lower, as determined by size exclusion chromatography (SEC). Conjugates

[0405] The conjugates described herein comprise at least one linker attached to an anti-Nectin-4 antibody or antigen-binding fragment thereof and at least one immunostimulatory compound, such as a myeloid cell agonist or other agonist (e.g., a TLR8 agonist, a TLR7 agonist, another TLR agonist, a STING agonist, a RIG-I-like receptor agonist, a c-type lectin receptor agonist, or a cytosolic DNA sensor agonist). In some embodiments, the present disclosure provides a conjugate comprising a conjugate of Formula I: [ka] (Wherein: A is an anti-Nectin-4 antibody or an antigen-binding fragment thereof, L is a linker; D x is an immunostimulatory compound; n is selected from 1 to 20; and z is selected from 1 to 20. The conjugate is represented by:

[0406] In some embodiments, the immunostimulatory compound is a myeloid cell agonist. In some embodiments, the immunostimulatory compound is a TLR8 agonist. In some embodiments, the immunostimulatory compound is a TLR7 agonist. In some embodiments, the immunostimulatory compound is a TLR3 agonist. In some embodiments, the immunostimulatory compound is a TLR4 agonist. In some embodiments, the immunostimulatory compound is a TLR5 agonist. In some embodiments, the immunostimulatory compound is a TLR9 agonist. In some embodiments, the immunostimulatory compound is a STING agonist. Exemplary STING agonist compounds include RG7854, ADU-S100, MK-1454, MK-2118, BMS-986301, GSK3745417, SB-11285, and IMSA-101. In some embodiments, the immunostimulatory compound is a RIG-I-like receptor agonist. In some embodiments, the immunostimulatory compound is a c-type lectin receptor agonist. In some embodiments, the immunostimulatory compound is a cytosolic DNA sensor agonist.

[0407] In some aspects, the present disclosure provides a conjugate comprising at least one immunostimulatory compound (e.g., a compound or a salt thereof), an anti-Nectin-4 antibody or antigen-binding fragment thereof, and at least one linker, wherein each immunostimulatory compound is linked, i.e., covalently bonded, to the anti-Nectin-4 antibody or antigen-binding fragment thereof through the linker. The linker can be selected from a cleavable linker or a non-cleavable linker. In some embodiments, the linker is cleavable. In other embodiments, the linker is non-cleavable. Linkers are further described in the present application in the preceding section, any one of which can be used to connect an antibody or antigen-binding fragment thereof to an immunostimulatory compound.

[0408] In a conjugate, drug loading is represented by z, where the number of immunostimulatory compound-linker molecules per antibody or the number of immunostimulatory compounds per antibody depends on the particular conjugate. Depending on the context, z can represent the average number of immunostimulatory compound (-linker) molecules per antibody, which also refers to the average drug loading. z can range from 1 to 20, 1 to 50, or 1 to 100. In some conjugates, z is preferably 1 to 8. In some preferred embodiments, when z represents the average drug loading, z ranges from about 2 to about 5. In some embodiments, z is about 2, about 3, about 4, or about 5. The average number of immunostimulatory compounds per antibody in a preparation of a conjugate may be characterized by conventional means, such as mass spectrometry, liquid chromatography / mass spectrometry (LC / MS), HIC, ELISA assay, and HPLC.

[0409] Some conjugates are consistent with the disclosure herein. The conjugates generally comprise an immunostimulatory compound covalently linked to an anti-Nectin-4 antibody or antigen-binding fragment thereof, which localizes the conjugate to a target tissue, cell population, or cell. The anti-Nectin-4 antibody or antigen-binding fragment thereof is covalently linked to the respective immunostimulatory compound, either directly or through a linker that connects the immunostimulatory compound to the anti-Nectin-4 antibody or antigen-binding fragment thereof. The anti-Nectin-4 antibodies or antigen-binding fragments thereof listed herein are similarly consistent with the conjugates disclosed herein.

[0410] Some exemplary conjugates are as follows: The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one immunostimulatory compound, and, optionally, at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR7 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR8 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one Category A compound (TLR8 agonist), and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one Category B compound (TLR7 agonist), and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one Category C compound (TLR8 agonist), and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR3 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR4 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR5 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one TLR9 agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one STING agonist, and at least one linker.The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one RIG-I agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one c-type lectin receptor agonist, and at least one linker. The conjugate can comprise an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, at least one cytosolic DNA sensor agonist, and at least one linker. Exemplary Conjugates

[0411] In certain embodiments, the present disclosure provides an immunostimulatory conjugate (or conjugates) of an anti-Nectin-4 antibody or antigen-binding fragment thereof and at least one compound of any one of formulas (IA), (IB), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IVA), (IVB), and (IVC) in Category A, each compound optionally coupled to the antibody or antigen-binding fragment via a linker. In certain embodiments, the present disclosure provides an immunostimulatory conjugate of an anti-Nectin-4 antibody or antigen-binding fragment thereof and at least one compound of any one of formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC) in Category B, each compound optionally coupled to the antibody or antigen-binding fragment via a linker. In certain embodiments, the present disclosure provides immunostimulatory conjugates of an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure and at least one compound of any one of Formulas Ia-Ih in Category C, wherein each compound is optionally coupled to the antibody or antigen-binding fragment via a linker. In certain embodiments, the average drug-to-antibody ratio (DAR) of a pharmaceutical composition comprising a conjugate of an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure is selected from 1 to about 8, 2 to about 6, about 3 to about 5, or about 4.

[0412] In certain embodiments, the present disclosure provides a pharmaceutical composition suitable for intravenous or subcutaneous administration, comprising an immunostimulatory compound of any one of Formulas (IA), (IB), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IVA), (IVB), and (IVC) of Category A conjugated to an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, and a pharmaceutically acceptable excipient. In certain embodiments, the present disclosure provides a pharmaceutical composition suitable for intravenous or subcutaneous administration, comprising an immunostimulatory compound of any one of Formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC) of Category B conjugated to an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, and a pharmaceutically acceptable excipient. In certain embodiments, the present disclosure provides a pharmaceutical composition suitable for intravenous or subcutaneous administration, comprising an immunostimulatory compound of any one of Formulas Ia-Ih of Category C conjugated to an anti-Nectin-4 antibody or antigen-binding fragment thereof of the present disclosure, and a pharmaceutically acceptable excipient. In certain embodiments, the average drug-to-antibody ratio (DAR) of the pharmaceutical composition of the conjugate is selected from 1 to about 8, 2 to about 6, about 3 to about 5, or about 4.

[0413] In certain embodiments, the present disclosure provides a method for treating a disease treatable by a TLR agonist (e.g., cancer), comprising subcutaneously administering an effective amount of a conjugate of a compound of any one of formulas (IA), (IB), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IVA), (IVB), and (IVC) of Category A, or a pharmaceutical composition thereof suitable for intravenous or subcutaneous administration to a subject in need thereof. In certain embodiments, the present disclosure provides a method for treating cancer (e.g., bladder, breast, lung, head and neck, cervix), comprising intravenously or subcutaneously administering an effective amount of a conjugate of a compound of any one of formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC) of Category B, or a pharmaceutical composition thereof suitable for subcutaneous administration to a subject in need thereof. In certain embodiments, the present disclosure provides a method for the treatment of cancer (e.g., bladder, breast, lung, head and neck, cervix) comprising intravenously or subcutaneously administering an effective amount of a conjugate of a compound of any one of Formulae Ia-Ih in Category C, or a pharmaceutical composition thereof suitable for subcutaneous administration, to a subject in need thereof. In any of the embodiments herein, the conjugate may be administered by slow infusion.

[0414] The present disclosure provides a compound of the formula: [ka] (In the formula: n is selected from 1 to 20; L 3 is a linker; D is selected from a compound or salt of any one of formulas (IA), (IB), (IIA), (IIB), (IIC), (IIIA), (IIIB), (IVA), (IVB), and (IVC) in Category A; formulas (IA), (IB), (IC), (IIA), (IIB), and (IIC) in Category B; and formulas (Ia), (Ib), (Ic), (Id), (Ie), (I...

Claims

1. An isolated monoclonal antibody or antigen-binding fragment thereof that specifically binds to Nectin-4, wherein the antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL); (1) The VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 1, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 2, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 3; and the VL comprises a CDR1 (VL-CDR1) comprising an amino acid sequence selected from any one of SEQ ID NOs: 4 to 6, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 7, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 8; (2) the VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 33, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 34, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 35; or the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 36, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 37, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 38; (3) the VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 39, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 40, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 41; or the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 42, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 43, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 44; (4) The VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 45, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 46, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 47; or the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 48, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 49, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 50; (5) The VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 51, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 52, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 53; and the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 54, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 55, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO:

56. or a VL-CDR3 comprising the amino acid sequence of (6) The VH comprises a CDR1 (VH-CDR1) comprising the amino acid sequence of SEQ ID NO: 57, a VH-CDR2 comprising the amino acid sequence of SEQ ID NO: 58, and a VH-CDR3 comprising the amino acid sequence of SEQ ID NO: 59; and the VL comprises a CDR1 (VL-CDR1) comprising the amino acid sequence of SEQ ID NO: 60, a VL-CDR2 comprising the amino acid sequence of SEQ ID NO: 61, and a VL-CDR3 comprising the amino acid sequence of SEQ ID NO: 62; An isolated monoclonal antibody or antigen-binding fragment thereof.

2. The antibody of claim 1, which is a humanized antibody.

3. (a) a VH comprising an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 10, and a VL comprising an amino acid sequence having at least 90% identity to an amino acid sequence selected from any one of SEQ ID NOs: 12-17; or (b) a VH comprising the amino acid sequence of SEQ ID NO: 10, and a VL comprising an amino acid sequence selected from any one of SEQ ID NOs: 12 to 17; The antibody of claim 1 or claim 2, comprising:

4. the VL-CDR1 comprises the amino acid sequence of SEQ ID NO:5; or the VL-CDR1 comprises the amino acid sequence of SEQ ID NO: 4; The antibody according to any one of claims 1 to 3.

5. The VL comprises the amino acid sequence of SEQ ID NO: 14; or the VL comprises the amino acid sequence of SEQ ID NO: 13; The antibody described in claim 4.

6. The antibody of any one of claims 1 to 5, comprising a VH comprising the amino acid sequence of SEQ ID NO:

10.

7. The antibody of any one of claims 1 to 6, comprising a constant region of human IgG1, human IgG2, human IgG3, or human IgG4.

8. (a) a heavy chain comprising an amino acid sequence that is at least 90% identical to the amino acid sequence of SEQ ID NO: 24, and a light chain comprising an amino acid sequence that has at least 90% identity to an amino acid sequence selected from any one of SEQ ID NOs: 26-31; or (b) a heavy chain comprising the amino acid sequence of SEQ ID NO: 24, and a light chain comprising an amino acid sequence selected from any one of SEQ ID NOs: 26-31. The antibody of any one of claims 1 to 7, comprising:

9. a heavy chain comprising the amino acid sequence of SEQ ID NO:24 and a light chain comprising the amino acid sequence of SEQ ID NO:28; or a heavy chain comprising the amino acid sequence of SEQ ID NO: 24, and a light chain comprising the amino acid sequence of SEQ ID NO: 27; The antibody according to any one of claims 1 to 8.

10. A conjugate comprising the antibody of any one of claims 1 to 9 and a small molecule drug.

11. A pharmaceutical composition comprising the antibody of any one of claims 1 to 9 or the conjugate of claim 10, and a pharmaceutically acceptable carrier.

12. An isolated nucleic acid encoding the amino acid sequence of the antibody of any one of claims 1 to 9.

13. A vector comprising the nucleic acid of claim 12.

14. 1. An isolated host cell comprising: (a) comprising a nucleic acid according to claim 12 or a vector according to claim 13; or (b) an isolated host cell expressing the antibody of any one of claims 1 to 9.

15. A method for producing an antibody that binds to Nectin-4, comprising: A method comprising culturing the host cell of claim 14 under conditions suitable for expressing the antibody.

16. 16. The method of claim 15, further comprising isolating the antibody.

17. A composition for use in a method for treating Nectin-4-expressing cancer, comprising the antibody of any one of claims 1 to 9, wherein an effective amount of the antibody of any one of claims 1 to 9 is administered to a subject with Nectin-4-expressing cancer.

18. A composition for use in a method for treating a Nectin-4-expressing cancer, comprising the conjugate of claim 10, wherein an effective amount of the conjugate of claim 9 is administered to a subject having a Nectin-4-expressing cancer.

19. 12. The pharmaceutical composition of claim 10 for use in a method for treating a Nectin-4-expressing cancer, wherein an effective amount of the pharmaceutical composition of claim 11 is administered to a subject having a Nectin-4-expressing cancer.

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