Methods of Treating Cancer Using Anti-CD22 Antibody-Oligonucleotide Conjugates

Abstract

The present disclosure provides methods for treating cancer using anti-CD22 antibody-oligonucleotide conjugates.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 63 / 343,506, filed May 18, 2022, which is incorporated by reference in its entirety herein.

[0002] Reference to Electronic Sequence Listing The contents of the electronic format of the sequence listing (186492001040seqlist.xml, size: 319,072 bytes, and creation date: May 3, 2023) are incorporated herein by reference in their entirety.

[0003] The present disclosure relates generally to methods for treating cancer using anti-CD22 antibody-oligonucleotide conjugates. [Background technology]

[0004] Immune-mediated cancer elimination requires the coordination of the innate immune system (e.g., dendritic cells, macrophages, myeloid-derived suppressor cells) and the adaptive immune system (i.e., B cells, cytotoxic T cells, regulatory T cells). Current gold-standard cancer treatments include administration of immune checkpoint blockade therapies (anti-PD-1, anti-CTLA-4), but these therapies are effective in only up to 30% of patients and can be associated with severe, even fatal, side effects in 10–30% of cancer patients. Moreover, such therapies act specifically to increase the activity of primarily T lymphocytes, thereby ignoring the importance of coordinated immune responses. Thus, there remains a need for safe and effective therapies that exploit the broad range of possible immune involvement in cancer elimination. Summary of the Invention

[0005] In one aspect, provided herein is a method of treating cancer in an individual, the method comprising administering to the individual a conjugate comprising an anti-CD22 antibody (Ab) and an immune modulatory oligonucleotide (P) at a dose of 0.1 mg / kg to 60 mg / kg, wherein the Ab comprises two antibody light chains, two antibody heavy chains, and two Q tag peptides (Q), each of the two Q tag peptides comprising the amino acid sequence RPQGFGPP (SEQ ID NO: 49), wherein one Q tag peptide is linked to the C-terminus of each of the two antibody heavy chains, and one of the two Q tag peptides is linked to the immune modulatory oligonucleotide via an amide bond with a glutamine residue of the Q tag peptide and a linker (L) shown in formula (A): [ka] During the ceremony, [ka] indicates the attachment point of Q to an antibody (Ab), each heavy chain of Ab comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 113, CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and CDR3 comprising the amino acid sequence of SEQ ID NO: 116, each light chain of Ab comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 117, CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 120 to 122, and linker L is [ka] wherein m is 24; [ka] indicates the point of attachment to the oligonucleotide P, [ka] indicates the point of attachment to the glutamine residue of the Q tag); oligonucleotide P has the structure: [ka] or [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, [ka] indicates the attachment point within the oligonucleotide, [ka] indicates the point of attachment to the linker L.

[0006] In some embodiments, the individual has an inoperable, locally advanced, metastatic, and / or recurrent solid tumor. In some embodiments, the individual has a cancer selected from the group consisting of cutaneous melanoma, breast cancer, ovarian cancer, colorectal cancer, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), head and neck squamous cell carcinoma (HNSCC), Merkel cell carcinoma, hepatocellular carcinoma (HCC), cervical squamous cell carcinoma, urothelial carcinoma, endometrial carcinoma, gastroesophageal adenocarcinoma, and cholangiocarcinoma.

[0007] In one aspect, provided herein is a method of treating cancer in an individual, the method comprising administering to the individual a conjugate comprising an anti-CD22 antibody (Ab) and an immune modulatory oligonucleotide (P), wherein the individual has an inoperable, locally advanced, metastatic, and / or recurrent solid tumor, wherein the Ab comprises two antibody light chains, two antibody heavy chains, and two Q tag peptides (Q), each of the two Q tag peptides comprising the amino acid sequence RPQGFGPP (SEQ ID NO:49), wherein one Q tag peptide is linked to the C-terminus of each of the two antibody heavy chains, and one of the two Q tag peptides is linked to the immune modulatory oligonucleotide via an amide bond with a glutamine residue of the Q tag peptide and a linker (L) of the formula shown in formula (A): [ka] During the ceremony, [ka] indicates the attachment point of Q to an antibody (Ab), each heavy chain of Ab comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 113, CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and CDR3 comprising the amino acid sequence of SEQ ID NO: 116, each light chain of Ab comprises CDR1 comprising the amino acid sequence of SEQ ID NO: 117, CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 120 to 122, and linker L is [ka] wherein m is 24; [ka] indicates the point of attachment to the oligonucleotide P, [ka] indicates the point of attachment to the glutamine residue of the Q tag); oligonucleotide P has the structure: [ka] or [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, [ka] indicates the attachment point within the oligonucleotide, [ka] indicates the point of attachment to the linker L.

[0008] In some embodiments, the individual has a cancer selected from the group consisting of cutaneous melanoma, breast cancer, ovarian cancer, colorectal cancer, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), head and neck squamous cell carcinoma (HNSCC), Merkel cell carcinoma, hepatocellular carcinoma (HCC), cervical squamous cell carcinoma, urothelial carcinoma, endometrial carcinoma, gastroesophageal adenocarcinoma, and cholangiocarcinoma. In some embodiments, the conjugate is administered to the individual at a dose of 0.1 mg / kg to 60 mg / kg.

[0009] In some embodiments according to any of the embodiments described herein, the individual has a solid tumor that has progressed on and / or is intolerant to standard therapy. In some embodiments, the conjugate is administered to the individual at a dose of 0.1, 0.3, 1.0, 3.0, 10.0, 30.0, or 60.0 mg / kg. In some embodiments, the conjugate is administered to the individual once every two weeks. In some embodiments, each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 120. In some embodiments, each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 121. In some embodiments, each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 122. In some embodiments, each heavy chain of the Ab comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 65. In some embodiments, each light chain of the Ab comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 73. In some embodiments, each light chain of the Ab comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 87. In some embodiments, each heavy chain of the Ab that comprises a Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179 or 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 181 or 182. In some embodiments, each heavy chain of the Ab that comprises a Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 181.In some embodiments, each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 182. In some embodiments, each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 181. In some embodiments, each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 182. In some embodiments, the individual is a human. In some embodiments, the conjugate is administered in a formulation comprising a concentration of 30 mg / mL of the conjugate, 20 mM citrate, 150 mM L-arginine, 50 mM NaCl, and 0.02% polysorbate 80 (w / v), and the formulation is at pH 6.5. In some embodiments, the conjugate is administered intravenously to the individual.

[0010] The present application can be understood by reference to the following detailed description in conjunction with the accompanying drawings. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 shows a schematic diagram of an exemplary antibody:CpG conjugate with engineered Q-tags (RPQGFGPP, SEQ ID NO:49) fused to the C-terminus (bearing DAR1) of each heavy chain. [Figure 2A] 1 shows the levels of cell surface CD22 expression on peripheral B cells in patients following treatment with anti-CD22-CpG conjugates. [Figure 2B] Shown are levels of cell surface CD22 expression on peripheral B cells from patients prior to day 1 treatment with anti-CD22-CpG conjugate at the doses indicated (Pre-C1D1), 3 hours after day 1 (far left 3 hours), day 2 (C1D2), day 8 (C1D8), 15 hours prior to administration of the second dose (i.e., pre-2nd cycle, C1D15), 3 hours after administration of the second dose (far right 3 hours), day 16 during the 2nd cycle (C2D16), and day 22 during the 2nd cycle, (C2D22). [Figure 3A]1 shows CD86 expression on naive B cells following treatment with anti-CD22-CpG conjugates in patients at the doses indicated. [Figure 3B] 1 shows CD86 expression on memory B cells following treatment with anti-CD22-CpG conjugates in patients at the doses indicated. [Figure 3C] FIG. 1 shows CD86 expression on naive B cells following treatment with anti-CD22-CpG conjugates in patients at the doses and time points indicated. [Figure 4] 1 shows the concentration of anti-CD22-CpG conjugate (TRAAC) over time in patients after administration of 0.1 mg / kg, 0.3 mg / kg, or 1 mg / kg of the conjugate. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The following description sets forth example methods, parameters, etc. It should be understood, however, that such description is not intended to limit the scope of the present disclosure, but rather is provided as a description of representative embodiments.

[0013] Provided herein are methods of treating a solid tumor comprising administering a conjugate comprising an anti-CD22 antibody and an immune modulatory oligonucleotide. In some embodiments, the cancer is an advanced or metastatic solid tumor. In some embodiments, the conjugate is administered to an individual at a dose of 0.1 mg / kg to 60 mg / kg.

[0014] Toll-like receptor 9 (TLR9), also called CD289, is a transmembrane and intracytoplasmic pattern recognition receptor that recognizes pathogen-associated molecular patterns (PAMPs) and participates in immune responses in the absence of pathogens. TLR9 is an important receptor expressed by immune cells, including dendritic cells (DCs), B lymphocytes, macrophages, natural killer cells, and other antigen-presenting cells. Activation of TLR9 triggers intracellular signaling cascades that result in activation, maturation, proliferation, and cytokine production in these immune cells, thus bridging and coordinating innate and adaptive immunity. Martinez-Campos et al., Viral Immunol. 2016, 30, 98-105; Notley et al., Sci. Rep. 2017, 7, 42204. Thus, the identification of novel TLR9 agonist compounds may activate both innate and adaptive immunity and serve as potential anticancer therapeutics.

[0015] Natural TLR9 agonists include unmethylated cytosine-guanine dinucleotide (CpG)-containing oligodeoxynucleotides (CpG ODNs). CpG ODNs are often susceptible to degradation in serum, and therefore the pharmacokinetics of CpG ODNs may be one of the limiting factors in their development as therapeutics. In addition, CpG ODNs exhibit heterogeneous tissue distribution that may increase PAMP-related toxicity. These drawbacks(s) can be overcome by conjugating CpG ODNs to targeting moieties for the treatment of specific diseases, e.g., conjugating to anti-CD22 antibodies for anti-cancer therapy. Indeed, CD22 is a transmembrane sialoglycoprotein receptor with restricted expression in B cells, including tumor-infiltrating B cells (TIBs), which form complexes with anti-CD22-CpG conjugates and internalize them, thereby delivering TLR9 agonists to stimulate anti-cancer immune responses. Preclinical mouse tumor models have shown that anti-CD22-CpG conjugates activate the TLR9 pathway in B cells, increasing immune activation, cytokine production, antigen presentation, and antitumor effects. However, the safety and efficacy of this drug compound in human cancer patients needs to be evaluated.

[0016] I. Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. All patents, applications, published applications and other publications mentioned herein are incorporated by reference in their entirety. If the definitions set forth in this section are contrary to or otherwise inconsistent with the definitions set forth in the patents, applications, or other publications incorporated by reference herein, the definitions set forth in this section shall take precedence over the definitions incorporated by reference herein.

[0017] It is understood that certain features of the present disclosure that are described for clarity in the context of separate embodiments can also be provided in combination in a single embodiment. Conversely, various features of the present disclosure that are described for brevity in the context of a single embodiment can also be provided separately or in any suitable subcombination. All combinations of embodiments relating to specific method steps, reagents, or conditions are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed.

[0018] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as a priori basis for the use of exclusive terminology such as "solely," "only," or the use of a "negative" limitation in connection with the recitation of claim elements.

[0019] Throughout this application, unless the context indicates otherwise, references to compounds of formula (A) include ionic forms, polymorphs, pseudopolymorphs, amorphous forms, solvates, cocrystals, chelates, isomers, tautomers, oxides (e.g., N-oxides, S-oxides), esters, prodrugs, isotopes, and / or protected forms thereof. In some embodiments, references to compounds of formula (A) include polymorphs, solvates, cocrystals, isomers, tautomers, and / or oxides thereof. In some embodiments, references to compounds of formula (A) include polymorphs, solvates, and / or cocrystals thereof. In some embodiments, references to compounds of formula (A) include isomers, tautomers, and / or oxides thereof. In some embodiments, references to compounds of formula (A) include solvates thereof.

[0020] As used herein, the term "immunomodulatory polynucleotide" or "immunomodulatory oligonucleotide" refers to a polynucleotide construct containing a total of 6 to 50 consecutive nucleosides. The immunomodulatory polynucleotide can modulate an innate immune response as determined, for example, by a change in the activity of an intracellular signaling pathway(s), including, but not limited to, NFκB, a change in the expression of an activation marker, or a change in the secretion of at least one inflammatory cytokine or at least one type I interferon in an immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has been delivered (compared to another immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has not been delivered) or in an immune cell that interacts with the immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has been delivered (direct cell-cell interaction and, for example, direct stimulation from one or more cytokines secreted from the cell to which the immunomodulatory polynucleotide has been delivered).

[0021] As used herein, the term "immunostimulatory polynucleotide" or "immunostimulatory oligonucleotide" refers to an immunomodulatory polynucleotide that is capable of activating an immune response as determined, for example, by an increase in the activity of an intracellular signaling pathway(s) such as NFκB, an increase in the level of a cell surface marker(s) of activation or function, or an increase in the secretion of at least one proinflammatory cytokine or at least one type I interferon in an immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has been delivered (e.g., compared to another immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has not been delivered), or in an immune cell that interacts with the immune cell (e.g., an antigen presenting cell) to which the immunomodulatory polynucleotide has been delivered (direct cell-cell interaction and, for example, direct stimulation from one or more cytokines secreted from the cell to which the immunomodulatory polynucleotide has been delivered).

[0022] It is to be understood that the terms "polynucleotide" and "oligonucleotide" may be used interchangeably herein. It will be further understood that the terms "immunomodulatory polynucleotide", "immunostimulatory polynucleotide", "immunosuppressive polynucleotide", and "conjugate" encompass salts of immunomodulatory polynucleotides, immunostimulatory polynucleotides, immunosuppressive polynucleotides, and conjugates, respectively. For example, ... - moieties, where X is O or S. Thus, R E1 , R E2 , and R E3It should be understood that the phosphates and phosphodiesters described as having one or more of include salts in which the phosphate, thiophosphate, or dithiophosphate is present in deprotonated ionic form. Additionally, the terms "free," "naked," and "unconjugated" may be used interchangeably herein to refer to immunomodulatory, immunostimulatory, or immunosuppressive polynucleotides and / or oligonucleotides (e.g., CpG oligonucleotides).

[0023] As used herein, the term "phosphate triester" refers to a phosphate ester in which all three valencies are replaced with non-hydrogen substituents. The phosphate triester comprises a phosphate, thiophosphate, or dithiophosphate; one or two bonds to a nucleoside(s) or abasic spacer(s), and / or phosphoryl group(s); and one or two groups independently selected from the group consisting of a bioreversible group; a non-bioreversible group; an auxiliary moiety; a conjugation group; a targeting moiety, and optionally a linker attached to one or more (e.g., 1-6) auxiliary moieties. The terminal phosphate triester comprises one bond to a nucleoside-containing group and two groups independently selected from the group consisting of a bioreversible group, a non-bioreversible group, an auxiliary moiety, a conjugation group, a phosphoryl group, and a targeting moiety, and optionally a linker attached to one or more (e.g., 1-6) auxiliary moieties. In some embodiments, the terminal phosphate triester contains one or zero linkers attached to a targeting moiety and, optionally, one or more (e.g., 1-6) auxiliary moieties. The internucleoside phosphate triester contains two bonds to a nucleoside-containing group. The phosphate triester comprises a group of the following structure: [ka] wherein X E1 and X E2 each is independently O or S; R E1 and R E3each of which is independently a nucleoside; a sugar analog of an abasic spacer; a bioreversible group; a non-bioreversible group; an ancillary moiety; a conjugation group; a linker attached to a targeting moiety; a linker attached to a targeting moiety and one or more (e.g., 1-6) ancillary moieties; or a group of formula -P(=X E1 )(-X E2 -R E2 A) a bond to the phosphorus atom in the -O- group, (In the formula, R E2 A is hydrogen; a bioreversible group; a non-bioreversible group; an auxiliary moiety; a conjugation group; a linker attached to a targeting moiety; or a linker attached to a targeting moiety and one or more (e.g., 1-6) auxiliary moieties; R E2 is a bioreversible group; a non-bioreversible group; an auxiliary moiety; a conjugation group; a linker attached to a targeting moiety; or a linker attached to a targeting moiety and one or more (e.g., 1 to 6) auxiliary moieties, However, R E1 and R E3 At least one of the is a bond to a group containing at least one nucleoside.

[0024] R E1 and R E3 When both R and R are bonds to a group that contains at least one nucleoside, the phosphate triester is an internucleoside phosphate triester. E1 and R E3 is the bond to the nucleoside-containing group, the phosphate triester is a terminal phosphate triester.

[0025] As used herein, the term "amino acid" refers to any amino acid (both standard and non-standard), including, but not limited to, α-amino acids, β-amino acids, γ-amino acids, and δ-amino acids. Examples of suitable amino acids include, but are not limited to, alanine, asparagine, aspartate, cysteine, glutamic acid, glutamine, glycine, proline, serine, tyrosine, arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine. Additional examples of suitable amino acids include, but are not limited to, ornithine, hypusine, 2-aminoisobutyric acid, dehydroalanine, gamma-aminobutyric acid, citrulline, beta-alanine, alpha-ethyl-glycine, alpha-propyl-glycine, and norleucine.

[0026] The terms "antibody," "immunoglobulin," and "Ig" are used interchangeably herein and are used in the broadest sense to specifically refer to, for example, individual monoclonal antibodies (including agonist, antagonist, neutralizing, full-length or intact monoclonal antibodies), antibody compositions having polyepitopic or monoepitopic specificity, polyclonal or monovalent antibodies, multivalent antibodies, multispecific antibodies formed from at least two intact antibodies (e.g., bispecific antibodies so long as they exhibit the desired biological activity), single chain antibodies, and fragments of antibodies. Antibodies may be human, humanized, chimeric, and / or affinity matured antibodies, as well as antibodies derived from other species, e.g., mouse and rabbit.

[0027] The term "antibody" is intended to include polypeptide products of B cells within the immunoglobulin class of polypeptides, capable of binding to a specific antigen, and composed of a pair of two identical polypeptide chains, each pair having one heavy chain (about 50-70 kDa) and one light chain (about 25 kDa), with the respective amino-terminal portions of each chain containing a variable region of about 100 to about 130 or more amino acids, and the respective carboxy-terminal portions of each chain containing a constant region. See Borrebaeck (ed.) (1995) Antibody Engineering, Second Ed., Oxford University Press.; Kuby (1997) Immunology, Third Ed., W.H. Freeman and Company, New York. Antibodies also include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinant antibodies, polyspecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, camelized antibodies, chimeric antibodies, intrabodies, anti-idiotypic (anti-Id) antibodies, and functional fragments thereof. A functional fragment refers to a portion of an antibody heavy or light chain polypeptide that retains some or all of the binding activity of the antibody from which the fragment is derived. Non-limiting examples of antibody functional fragments include single chain Fvs (scFvs) (e.g., including monospecific or bispecific), Fab fragments, F(ab') fragments, F(ab) 2 Fragment, F(ab') 2Fc variants include Fc variants, disulfide-linked Fv (sdFv), Fd fragments, Fv fragments, scRv-Fc, nanobodies, diabodies, triabodies, tetrabodies, and minibodies. In some embodiments, the antibodies include Fc variants with reduced or eliminated effector functions. In particular, the antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, such as antigen binding domains or molecules that contain an antigen binding site that binds an antigen (e.g., one or more complementarity determining regions (CDRs) of an anti-CD56 antibody or an anti-SIRPα antibody). Such antibody fragments are described, for example, in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York (1989); Myers (ed.), Molec. Biology and Biotechnology: A Comprehensive Desk Reference, New York: VCH Publisher, Inc.; Huston et al., Cell Biophysics 1993, 22, 189-224; Pluckthun and Skerra, Meth. Enzymol. 1989, 178, 497-515; and Day, Advanced Immunochemistry, Second Ed., Wiley-Liss, Inc., New York, NY (1990). The antibodies provided herein can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG2a and IgG2b) of immunoglobulin molecule.

[0028] The term "antigen" refers to a predetermined target to which an antibody can selectively bind. A target antigen can be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or fragment thereof, or other naturally occurring or synthetic compound. In one embodiment, the target antigen is a polypeptide.

[0029] The terms "antigen-binding fragment," "antigen-binding domain," and "antigen-binding region" refer to the portion of an antibody that contains the amino acid residues that interact with an antigen (e.g., a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or fragment thereof, or other naturally occurring or synthetic compound) and confer upon the binder its specificity and affinity for the antigen (e.g., the complementarity determining region (CDR)).

[0030] The terms "specific binding," "specifically binds to," or "specific for" a particular polypeptide or epitope on a particular polypeptide target means, for example, at least about 10 -4 M, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10 -9 M, at least about 10 -10 M, at least about 10 -11 M, or at least about 10 -12 Dissociation constant (K d In one embodiment, the term "specific binding" refers to binding by a molecule to a particular polypeptide or an epitope on a particular polypeptide without substantially binding to other polypeptides or polypeptide epitopes.

[0031] The four-chain antibody unit is a heterotetrameric glycoprotein consisting of two identical light (L) chains and two identical heavy (H) chains. For IgG, the four-chain unit is generally about 150,000 daltons. Each L chain is linked to the H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the isotype of the H chain. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at the N-terminus, followed by three constant domains (CH) for each of the α and γ chains, and four CH domains for the μ and ε isotypes. Each L chain has a variable domain (VL) at the N-terminus, followed by a constant domain (CL) at its opposite end. The VL is aligned with the VH, and the CL is aligned with the first constant domain (CH1) of the heavy chain. Certain amino acid residues are believed to form an interface between the light and heavy chain variable domains. A VH and a VL pair together to form a single antigen-binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology, 8th edition, Stites et al. (eds.), Appleton & Lange, Norwalk, CT, 1994, page 71 and Chapter 6.

[0032] The term "variable region" or "variable domain" refers to a portion of an antibody's light or heavy chain, generally located at the amino terminus of the light or heavy chain, about 120-130 amino acids long for heavy chains and about 100-110 amino acids long for light chains, that is used in the binding and specificity of each particular antibody to a particular antigen. The variable region of the heavy chain may be referred to as "VH". The variable region of the light chain may be referred to as "VL". The term "variable" refers to the fact that certain segments of the variable region vary in sequence extensively between antibodies. The V region mediates antigen binding and determines the specificity of a particular antibody to its particular antigen. However, the variability is not evenly distributed across the 110 amino acid span of the variable region. Instead, the V region consists of less variable (e.g., relatively invariant) stretches of about 15-30 amino acids called framework regions (FR), separated by shorter regions of greater variability (e.g., extreme variability) called "hypervariable regions", each about 9-12 amino acids long. The variable regions of the heavy and light chains each contain four FRs, mainly in a β-sheet structure, connected by three hypervariable regions, which form loops connecting and sometimes forming part of the β-sheet structure. The hypervariable regions in each chain are held in close proximity to the hypervariable regions of the other chain by the FRs, and contribute to the formation of the antigen-binding site of the antibody (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991). The constant regions are not directly involved in the binding of the antibody to the antigen, but are involved in various effector functions, such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) of the antibody. The variable regions vary widely in sequence between different antibodies. The sequence variability is concentrated in the CDRs, while the less variable parts of the variable regions are called framework regions (FRs). The CDRs of the light and heavy chains are primarily responsible for the interaction of the antibody with the antigen. In certain embodiments, the variable region is a human variable region.

[0033] The term "Kabat variable region residue numbering" or "Kabat amino acid position numbering", and variations thereof, refers to the numbering system used for the heavy or light chain variable regions of an antibody sequence in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortening of or insertion into the FRs or CDRs of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 of H2 (residue 52a according to Kabat) and may also contain multiple inserted residues after heavy chain FR residue 82 (e.g., residues 82a, 82b and 82c according to Kabat, etc.). The Kabat numbering of residues may be determined for a particular antibody by alignment of the antibody's sequence with the "standard" Kabat numbered sequence at the regions of homology. The Kabat numbering system is commonly used when referring to residues in the variable domain (corresponding roughly to residues 1-107 in the light chain and residues 1-113 in the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The "EU numbering system" or "EU index" is commonly used when referring to residues in the immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al, supra). "EU index according to Kabat" refers to the numbering of residues in a human IgG1 EU antibody. Other numbering schemes are described, for example, by AbM, Chothia, Contact, IMGT, and AHon.

[0034] An "intact" antibody is one that comprises an antigen-binding site, as well as a CL and at least a heavy chain constant region, CH1, CH2, and CH3. The constant region may comprise a human constant region or an amino acid sequence variant thereof. Preferably, the intact antibody has one or more effector functions.

[0035] The term "antibody fragment" refers to a portion of an intact antibody, preferably the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies and di-diabodies (see, e.g., Holliger et al., Proc. Natl. Acad. Sci. USA 1993, 90, 6444-8; Lu et al., J. Biol. Chem. 2005, 280, 19665-72; Hudson et al., J. Immunol. 2006, 271, 1997; Hudson et al., J. Immunol. 2005, 271, 1997). al., Nat. Med. 2003, 9, 129-134; WO 93 / 11161; and U.S. Pat. Nos. 5,837,242 and 6,492,123); single chain antibody molecules (see, e.g., U.S. Pat. Nos. 4,946,778; 5,260,203; 5,482,858, and 5,476,786); dual variable domain antibodies (see, e.g., U.S. Pat. No. 7,612,181), single variable domain antibodies (sdAbs) (see, e.g., Woolven et al., Immunogenetics 1999, 50, 98-101; Streltsov et al., Proc. Natl. Acad. Sci. USA 2004, 101, 12444-12449); and multispecific antibodies formed from antibody fragments.

[0036] The terms "functional fragment," "binding fragment," or "antigen-binding fragment" of an antibody refer to a molecule that exhibits at least one of the biological functions attributed to an intact antibody, which function includes at least binding to a target antigen.

[0037] The term "heavy chain", when used in reference to an antibody, refers to a polypeptide chain of about 50-70 kDa, the amino terminal portion containing a variable region of about 120-130 or more amino acids, and the carboxyl terminal portion containing a constant region. The constant region can be one of five different types (e.g., isotypes), called alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the heavy chain constant region. The different heavy chains differ in size; i.e., α, δ, and γ contain about 450 amino acids, and μ and ε contain about 550 amino acids. When combined with light chains, these different types of heavy chains give rise to the five well-known classes (e.g., isotypes) of antibodies, namely, IgA, IgD, IgE, IgG, and IgM, respectively, with four subclasses of IgG, namely, IgG1, IgG2, IgG3, and IgG4. The heavy chain can be a human heavy chain.

[0038] The term "light chain" when used in reference to an antibody refers to a polypeptide chain of about 25 kDa, the amino terminal portion of which contains a variable region of about 100 to about 110 or more amino acids, and the carboxyl terminal portion of which contains a constant region. The approximate length of a light chain is 211 to 217 amino acids. Based on the amino acid sequence of the constant domain, there are two different types, called kappa (κ) or lambda (λ). Light chain amino acid sequences are well known in the art. The light chain may be a human light chain.

[0039] As used herein, the term "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies constituting the population are identical except for possible naturally occurring mutations that may be present in minor amounts, and each monoclonal antibody typically recognizes a single epitope of an antigen. In certain embodiments, a "monoclonal antibody" as used herein is an antibody produced by a single hybridoma or other cell, and the antibody binds only to a beta-cloto, as determined, for example, by ELISA or other antigen-binding or competitive binding assays well known in the art. The term "monoclonal" is not limited to any particular method for making the antibody. For example, monoclonal antibodies useful in the present disclosure may be prepared by the hybridoma method first described by Kohler et al., Nature 1975, 256, 495, or may be made using recombinant DNA methods in bacterial, eukaryotic animal or plant cells (see, e.g., U.S. Pat. No. 4,816,567). "Monoclonal antibodies" can also be isolated from phage antibody libraries using techniques described, for example, in Clackson et al., Nature 1991, 352, 624-628 and Marks et al., J. Mol. Biol. 1991, 222, 581-597. Other methods for preparing clonal cell lines and the monoclonal antibodies expressed thereby are well known in the art (see, for example, Chapter 11 in: Short Protocols in Molecular Biology, (2002) 5th Ed., Ausubel et al., eds., John Wiley and Sons, New York). Exemplary methods for producing monoclonal antibodies are provided in the Examples herein.

[0040] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies comprising a human immunoglobulin (e.g., recipient antibody) in which native CDR residues are replaced by residues from the corresponding CDRs of a non-human species, e.g., mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and capacity (e.g., donor antibody). In some cases, residues from one or more FR regions of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may contain residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further refine antibody performance. The heavy or light chains of the humanized antibody can comprise substantially all of at least one or more variable regions, in which all or substantially all of the CDRs correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of human immunoglobulin sequences. In certain embodiments, a humanized antibody comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 1986, 321, 522-525; Riechmann et al., Nature 1988, 332, 323-329; Presta, Curr. Opin. Biotechnol. 1992, 3, 394-398; Carter et al., Proc. Natl. Acad. Sci. USA 1992, 89, 4285-4289; and U.S. Patent Nos. 6,800,738, 6,719,971, 6,639,055, 6,407,213, and 6,054,297.

[0041] A "human antibody" is an antibody having an amino acid sequence that corresponds to that of an antibody produced by a human and / or an antibody produced using any of the techniques for producing human antibodies as disclosed herein. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques well known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol. 1991, 227, 381; Marks et al., J. Mol. Biol. 1991, 222, 581) and yeast display libraries (Chao et al., Nature Protocols 2006, 1, 755-768). For the production of human monoclonal antibodies, the methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boerner et al., J. Immunol. 1991, 147, 86-95 can also be used. Also see van Dijk and van de Winkel, Curr. Opin. Pharmacol. 2001, 5, 368-374. Human antibodies can be prepared by administering antigen to transgenic animals, e.g., mice, whose endogenous loci have been disabled but which have been engineered to produce such antibodies in response to antigenic challenge (see, e.g., Jakobovits, Curr. Opin. Biotechnol. 1995, 6, 561-566; Bruggemann and Taussing, Curr. Opin. Biotechnol. 1997, 8, 455-458; and U.S. Pat. Nos. 6,075,181 and 6,150,584 for XENOMOUSE™ technology). Also see, e.g., Li et al., Proc. Natl. Acad. Sci. USA 2006, 103, 3557-3562 for human antibodies produced by human B cell hybridoma technology.

[0042] "CDR" refers to one of the three hypervariable regions (H1, H2 or H3) in the non-framework region of the β-sheet framework of the VH of an immunoglobulin (Ig or antibody), or one of the three hypervariable regions (L1, L2 or L3) in the non-framework region of the β-sheet framework of the VL of said antibody. Thus, CDRs are variable region sequences interspersed within the sequences of the framework regions. CDR regions are well known to those skilled in the art and are defined, for example, by Kabat as the most hypervariable regions within the antibody variable (V) domain. Kabat et al., J. Biol. Chem. 1977, 252, 6609-6616; Kabat, Adv. Protein Chem. 1978, 32, 1-75. The sequences of CDR regions are also structurally defined by Chothia as residues that are not part of the conserved β-sheet framework and therefore can adopt different conformations. Chothia and Lesk, J. Mol. Biol. 1987, 196, 901-917. Both terms are well recognized in the art. The sequences of the CDR regions are also defined by AbM, Contact, and IMGT. The positions of the CDRs in standard antibody variable regions have been identified by comparison of multiple structures. Al-Lazikani et al., J. Mol. Biol. 1997, 273, 927-948; Morea et al., Methods. 2000, 20, 267-279. Because the number of residues in hypervariable regions varies from antibody to antibody, additional residues for the standard positions are conventionally numbered a, b, c, etc., next to the residue number in the standard variable region numbering scheme. Al-Lazikani et al., supra (1997). Such nomenclature is likewise well known to those skilled in the art.

[0043] As used herein, the term "hypervariable region", "HVR" or "HV" refers to the region of an antibody variable region that is hypervariable in sequence and / or forms structurally distinct loops. Generally, antibodies contain six hypervariable regions, three in the VH (H1, H2, H3) and three in the VL (L1, L2, L3). Multiple hypervariable region delineations are in use and are encompassed herein. The Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia instead refers to the location of structural loops. See, e.g., Chothia and Lesk, J. Mol. Biol. 1987, 196, 901-917. When numbered using the Kabat numbering convention, the end of the Chothia CDR-H1 loop varies between H32 and H34 depending on the length of the loop (this is because the Kabat numbering scheme places the insertion at H35A and H35B, so that if neither 35A nor 35B are present, the loop ends at 32, if only 35A is present, the loop ends at 33, and if both 35A and 35B are present, the loop ends at 34). The AbM hypervariable regions represent the intermediate Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software (see, e.g., Martin, in Antibody Engineering, Vol. 2, Chapter 3, Springer Verlag). The "contact" hypervariable regions are based on an analysis of the available complex crystal structures. Residues from each of these hypervariable regions or CDRs are shown below.

[0044] The term "Fc region" herein is used to define the C-terminal region of an immunoglobulin heavy chain, including, for example, native sequence Fc regions, recombinant Fc regions, and variant Fc regions. Although the boundaries of an immunoglobulin heavy chain Fc region may vary, the human IgG heavy chain Fc region is usually defined as extending from amino acid residue position Cys226 or Pro230 to its carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) may be removed, for example, during antibody production or purification, or by genetically engineering the nucleic acid encoding the antibody heavy chain. Thus, a composition of intact antibodies may include an antibody population with all K447 residues removed, an antibody population with no K447 residues removed, and an antibody population with a mixture of antibodies with and without the K447 residue.

[0045] The term "DAR" refers to the drug-antibody ratio of an oligonucleotide-antibody conjugate, more specifically, the immunomodulatory polynucleotide-antibody ratio.In some cases, for example, an oligonucleotide-antibody conjugate can be described herein as having a DAR of 1, or as a DAR1 conjugate, where the oligonucleotide-antibody ratio is 1:1.In other cases, for example, an oligonucleotide-antibody conjugate can be described herein as having a DAR of 2, or as a DAR2 conjugate, where the oligonucleotide-antibody ratio is 2:1.

[0046] As used herein, the terms "including," "containing," and "comprising" are used in their open, non-limiting sense, and it is also understood that aspects and embodiments of the invention described herein include aspects and embodiments "consisting of" and / or "consisting essentially of."

[0047] It is understood that all quantities provided herein, whether the term "about" is expressly used or not, are meant to refer to the actual given value, which also refers to approximations to such given value that would be reasonably inferred based on ordinary skill in the art, including equivalences and approximations by experimental and / or measurement conditions for such given value.

[0048] As used herein, "carrier" includes pharma- ceutically acceptable carriers, excipients, or stabilizers that are non-toxic to cells or mammals exposed thereto at the dosages and concentrations used. Often, physiologically acceptable carriers are pH-buffered aqueous solutions. Non-limiting examples of physiologically acceptable carriers include buffers such as phosphate, citric acid, and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents, such as EDTA; sugar alcohols, such as mannitol or sorbitol; salt-forming counterions, such as sodium; and / or non-ionic surfactants, such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0049] As used herein, the term "effective amount" or "therapeutically effective amount" of a substance is at least the minimum concentration required to bring about a measurable improvement (e.g., toward a therapeutic endpoint) or prevention of a particular disorder. The effective amount herein may vary depending on factors such as the patient's medical condition, age, sex, and weight, as well as the ability of the substance to elicit a desired response in an individual. An effective amount is also one in which the therapeutically beneficial effects outweigh any toxic or adverse effects of the treatment. In the context of cancer, an effective amount includes an amount sufficient to cause tumor shrinkage and / or reduce the rate of tumor growth (e.g., suppress tumor growth), or prevent or delay other undesirable cell proliferation in cancer, or reach a particular threshold of response criteria. In some embodiments, an effective amount is an amount sufficient to delay the onset of cancer. In some embodiments, an effective amount is an amount sufficient to prevent or delay recurrence. In some embodiments, an effective amount is an amount sufficient to reduce the recurrence rate in an individual. An effective amount can be administered in one or more administrations. An effective amount of a drug or composition may (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, suppress, slow to some extent, preferably stop, the invasion of cancer cells into peripheral organs; (iv) inhibit (i.e., slow to some extent, preferably stop) tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay tumor onset and / or recurrence; (vii) reduce the rate of tumor recurrence; and / or (viii) alleviate to some extent one or more of the symptoms associated with cancer. An effective amount may be administered in one or more administrations. For purposes of this disclosure, an effective amount of a drug, compound, or pharmaceutical composition is an amount sufficient to directly or indirectly achieve a prophylactic or therapeutic treatment. As will be understood in a clinical context, an effective amount of a drug, compound, or pharmaceutical composition may or may not be obtained in combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in relation to the administration of one or more therapeutic agents, and a single agent may be considered to be administered in an effective amount when a desired result may or is achieved in combination with one or more other agents.

[0050] "Package insert" refers to instructions customarily included in commercial packaging of a drug, containing information regarding indications, usage, dosage, administration, contraindications, information regarding other drugs that may be combined with the packaged product, and / or warnings regarding the use of such drugs.

[0051] The terms "protein," "polypeptide," and "peptide" are used herein to refer to polymers of amino acids of any length. The polymers may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. These terms also encompass amino acid polymers that are modified naturally or by intervention, e.g., disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, e.g., conjugation with a labeling moiety. Typically, a protein for use herein has a molecular weight of at least about 5-20 kDa, alternatively at least about 20-100 kDa, or at least about 100 kDa. For example, proteins containing one or more analogs of an amino acid (including, e.g., unnatural amino acids, etc.), as well as other modifications known in the art, are also included in this definition.

[0052] A "pharmacologically acceptable salt" is a salt form that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject. See generally, Berge et al. (1977) J.Pharm.Sci. 66, 1. A particular pharma-ceutically acceptable salt is one that is pharmacologically effective and suitable for contact with the tissues of a subject without undue toxicity, irritation, or allergic reaction. Pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids, such as acetic acid, oxalic acid, propionic acid, succinic acid, maleic acid, tartaric acid, and the like. These salts may be derived from inorganic or organic acids. Non-limiting examples of pharma- ceutically acceptable salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne, hexyne-1,4-dioate ... The salts include, but are not limited to, 1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, methylsulfonate, propylsulfonate, besylate, xylenesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, and mandelate. In some embodiments, pharma- ceutically acceptable salts are formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or when coordinated with an organic base.Salts derived from pharma-ceutically acceptable organic non-toxic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, naturally occurring substituted amines, cyclic amines, and substituted amines including basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, tromethamine, trimethanine, dicyclohexylamine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-ethylglucamine, N-methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, amino acids such as lysine, arginine, histidine, etc. Examples of pharma-ceutically acceptable base addition salts include those derived from inorganic bases, such as sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. In some embodiments, organic non-toxic bases are L-amino acids, such as L-lysine and L-arginine, tromethamine, N-ethylglucamine and N-methylglucamine. Acceptable inorganic bases include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like. Lists of other suitable pharma-ceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 17th Edition, Mack Publishing Company, Easton, Pa., 1985.

[0053] A "solvate" is formed by the interaction of a solvent with a compound. Suitable solvents include, for example, water and alcohols (e.g., ethanol). Solvates include hydrates of any ratio of water to the compound, such as monohydrates, dihydrates, and hemihydrates.

[0054] A "subject," "patient," or "individual" is a human. An individual can be male or female and of any appropriate age, including infant, juvenile, adolescent, adult, and geriatric subjects.

[0055] The term "cancer" or "tumor" refers to the presence of cells that possess typical characteristics of cancer-causing cells, such as uncontrolled proliferation, immortality, metastatic potential, rapid growth and proliferation rate, and certain characteristic morphological features. Cancer cells are often in the form of solid tumors, which are detectable by procedures such as CAT scan, MR imaging, X-ray, ultrasound, or tactile based on tumor mass, and / or by the expression of one or more cancer-specific antigens in samples obtained from patients. In some embodiments, solid tumors do not have to have a measurable size. Cancer cells can also be in the form of liquid tumors, which may be alone or disseminated within an animal. As used herein, the terms "disseminated tumors" and "liquid tumors" are used interchangeably and include, but are not limited to, leukemia and lymphoma, as well as other blood cell cancers.

[0056] The terms "cancer recurrence" and "cancer relapse" are used interchangeably and refer to the return of signs, symptoms, or disease after remission. Recurrent cancer cells may reappear at the same site of the primary tumor or at another location, such as a secondary cancer. Cancer cells may reappear with the same disease form as the primary cancer or with a different disease form. For example, in some embodiments, the primary cancer is a solid tumor and the recurrent cancer is a liquid tumor. In other embodiments, the primary cancer is a liquid tumor and the recurrent cancer is a solid tumor. In still other embodiments, the primary cancer and the recurrent cancer are both solid tumors or both liquid tumors. In some embodiments, the recurrent tumor expresses at least one tumor-associated antigen that is also expressed by the primary tumor.

[0057] The term "refractory cancer" as used herein refers to a cancer that does not respond to treatment, e.g., a cancer that is resistant at the start of treatment (e.g., treatment with immunotherapy) or that may become resistant during treatment. The terms "respond", "response", or "responsiveness" refer to an anti-cancer response, e.g., in the sense of a reduction in tumor size or inhibition of tumor growth. These terms can also refer to an improved prognosis, as reflected, for example, by an increase in time to recurrence (time to first recurrence sensing as a second primary cancer as a first event, or death without evidence of recurrence) or an increase in overall survival (time from treatment to death from any cause). Respond or have a response means that a beneficial endpoint is achieved upon exposure to a stimulus. Alternatively, negative or adverse symptoms are minimized, alleviated, or attenuated upon exposure to the stimulus. It will be understood that assessing the likelihood that a tumor or subject will respond favorably is equivalent to assessing the likelihood that a tumor or subject will not respond favorably (i.e., exhibit a lack of response or be non-responsive).

[0058] As used herein, cancer includes, but is not limited to, melanoma, breast cancer, lung cancer, bronchial cancer, colorectal cancer, prostate cancer, pancreatic cancer, gastric cancer, ovarian cancer, bladder cancer, brain or central nervous system cancer, peripheral nervous system cancer, esophageal cancer, cervical cancer, endometrial cancer, oral or pharyngeal cancer, liver cancer, kidney cancer, testicular cancer, biliary tract cancer, small intestine or appendix cancer, salivary gland cancer, thyroid cancer, adrenal cancer, osteosarcoma, chondrosarcoma, cancer of the blood tissue, B cell cancer, such as multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain diseases, such as alpha chain disease, gamma chain disease, and mu chain disease, benign monoclonal thrombosis, and immune cell amyloidosis. Other non-limiting examples of cancer types amenable to methods encompassed by the present invention include human sarcomas and carcinomas, such as fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, myeloma, angiosarcoma, endothelial tumor, lymphangiosarcoma, lymphangioendothelial tumor, synovoma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous adenocarcinoma, papillary carcinoma, papillary adenocarcinoma, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, liver cancer, choriocarcinoma, somatic cell tumor, embryonal tumor, Werm's tumor, cervical cancer, bone cancer, Brain tumors, testicular cancer, lung cancer, small cell lung carcinoma, bladder cancer, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma, retinoblastoma; leukemias, such as acute lymphocytic leukemia and acute myelocytic leukemia (myeloblastic, promyelocytic, myelomonocytic, monocytic, and erythroid); chronic leukemias (chronic myelocytic (granulocytic) leukemia and chronic lymphocytic leukemia), and polycythemia vera, lymphomas (Hodgkin's disease and non-Hodgkin's disease), multiple myeloma, Waldenstrom's macroglobulinemia, and heavy chain disease. In some embodiments, the cancer is epithelial in nature, including, but not limited to, bladder cancer, breast cancer, cervical cancer, colon cancer, gynecological cancer, renal cancer, laryngeal cancer, lung cancer, oral cancer, head and neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, or skin cancer. In other embodiments, the cancer is breast cancer, colorectal cancer, ovarian cancer, or lung cancer.In yet other embodiments, the epithelial cancer is non-small cell lung cancer, non-papillary renal cell carcinoma, cervical cancer, ovarian cancer (e.g., high-grade serous ovarian cancer), or breast cancer. Epithelial cancers can be characterized in a variety of other ways, including, but not limited to, serous, endometrial, mucinous, clear cell, Brenner, or undifferentiated.

[0059] The term "cancer therapy" or "cancer therapeutic agent" as used herein refers to a therapy or agent that can exert an antitumor effect or have antitumor activity. Such antitumor effect or activity can be shown as a decrease in the rate of tumor cell proliferation, viability, or metastatic activity. A possible way to show antitumor activity is to show a decrease in the rate of proliferation of abnormal cells that occurs during treatment, or a stable or reduced tumor size. Such activity can be evaluated using accepted in vitro or in vivo tumor models, including but not limited to xenograft models, allograft models, MMTV models, and other known models well known in the art for investigating antitumor activity.

[0060] The terms "treat", "treating" and "treatment" are meant to encompass alleviating or abrogating a condition, disorder, or disease, or one or more symptoms associated with the condition, disorder, or disease; or reducing or eradicating the cause(s) of the condition, disorder, or disease itself. In some embodiments, treating cancer results in a complete response (CR) or partial response (PR). In some embodiments, treating cancer results in a CR, PR, or stable disease (SD). Exemplary CR, PR, and SD criteria are provided herein.

[0061] As used herein, "prevent", "preventing" and "prevention" refer to a method of delaying and / or eliminating the onset of a condition, disorder or disease and / or its attendant symptoms; a method of preventing a subject from acquiring a condition, disorder or disease, or a method of reducing the risk that a subject will acquire a condition, disorder or disease.

[0062] "Optional" or "optionally" means that the event or circumstance described thereafter may or may not occur, and the description includes both cases where the event or circumstance occurs and cases where it does not occur. For example, "optionally substituted alkyl" encompasses both "alkyl" and "substituted alkyl" as defined herein. With respect to any group containing one or more substituents, it will be understood by those skilled in the art that such groups are not intended to introduce any substitution or substitution pattern that is sterically impractical, synthetically unfeasible, and / or inherently unstable. It will also be understood that when a group or moiety is optionally substituted, the present disclosure includes both embodiments in which the group or moiety is substituted and embodiments in which the group or moiety is unsubstituted.

[0063] The term "Q tag" as used herein means -NH 2It refers to a portion of a polypeptide that contains a glutamine residue that upon transglutaminase-mediated reaction with a compound containing an amine provides a conjugate containing the portion of the polypeptide, the glutamine residue containing a side chain modified to include an amide attached to the compound. Q-tags are well known in the art. Non-limiting examples of Q-tags are LLQGG (SEQ ID NO: 172), GGGLLQGG (SEQ ID NO: 173), RPQGF (SEQ ID NO: 47), and RPQGFGPP (SEQ ID NO: 49). In some embodiments, the Q-tag is attached to the C-terminus of the heavy chain of the antibody. In some embodiments, the Q-tag is attached to the light chain of the antibody. In some embodiments, the Q-tag is naturally occurring. For example, a mutation from N297 to N297A exposes Q295 of the antibody, where conjugation may occur (numbering according to the EU index as listed, for example, in Edelman, GM et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat, EA et al., Sequences of proteins of immunological interest. 5th Edition-US Department of Health and Human Services, NIH publication n° 91-3242, pp 662, 680, 689 (1991)). In some embodiments, the Q tag is within the Fc domain of the antibody.

[0064] II. Cancer Treatment Methods Provided herein is a method of treating cancer in an individual comprising administering a conjugate comprising an anti-CD22 antibody and an immune modulatory oligonucleotide (e.g., a CpG oligonucleotide). In some embodiments, the cancer is a tumor, e.g., an advanced or metastatic solid tumor. In some embodiments, the tumor is an inoperable, locally advanced, metastatic, or recurrent histologically or cytologically proven solid tumor. In some embodiments, the conjugate is administered to an individual with a solid tumor, and the cancer in the individual is advanced, or the individual has a cancer that is intolerant to standard treatments or for which standard treatments are unavailable. In some embodiments, the cancer being treated is cutaneous melanoma, breast cancer, ovarian cancer, colorectal cancer, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), head and neck squamous cell carcinoma (HNSCC), Merkel cell carcinoma, hepatocellular carcinoma (HCC), cervical squamous cell carcinoma, urothelial carcinoma, endometrial cancer, gastroesophageal adenocarcinoma, or cholangiocarcinoma. In some embodiments, the cancer being treated is triple negative breast cancer, melanoma, ovarian cancer, or colon cancer. In some embodiments, tumor tissue from the cancer being treated is tested for the presence or level of PD-L1. In some embodiments, the conjugate is administered to the individual at a dose of 0.1 mg / kg to 60 mg / kg (e.g., 0.1, 0.3, 1.0, 3.0, 10.0, 30.0, or 60.0 mg / kg). In some embodiments, the conjugate is administered to the individual at a dose of 0.1 to 1 mg / kg, 1 to 10 mg / kg, 10 to 30 mg / kg, or 30 to 60 mg / kg. In some embodiments, the dose administered is 0.1 mg / kg. In some embodiments, the dose administered is 0.3 mg / kg. In some embodiments, the dose administered is 1.0 mg / kg. In some embodiments, the dose administered is 3.0 mg / kg. In some embodiments, the dose administered is 10.0 mg / kg. In some embodiments, the dose administered is 30.0 mg / kg. In some embodiments, the dose administered is 60.0 mg / kg. In some embodiments, the dose is administered Q1W (weekly), or about once every 7 days.In some embodiments, the dose is administered Q2W (every other week) or about once every 14 days. In some embodiments, the dose is administered Q3W (every 3 weeks) or about once every 21 days. In some embodiments, the dose is administered Q4W (every 4 weeks), about once every 28 days, or about once a month. In some embodiments, the dose is administered to the individual once, twice, three times, four times, five times, or more than five times during the course of treatment. In some embodiments, the individual is a human. In some embodiments, the conjugate is administered intravenously (IV) to the individual. In some embodiments, the conjugate is administered in a formulation comprising a concentration of 30 mg / mL of the conjugate, 20 mM citrate, 150 mM L-arginine, 50 mM NaCl, and 0.02% polysorbate 80 (w / v), and the formulation is at pH 6.5.

[0065] In some embodiments, treatment with the methods of the present disclosure results in a complete response (CR) in an individual. In some embodiments, CR refers to the disappearance of all non-nodal target lesions and a decrease of less than 10 mm in the short axis of any pathological lymph node assigned as a target lesion.

[0066] In some embodiments, treatment with the disclosed methods results in a partial response (PR) in an individual. In some embodiments, PR refers to at least a 30% reduction in the sum of the diameters of all target lesions relative to the sum of the diameters at baseline.

[0067] In some embodiments, treatment with the disclosed method results in no change (SD) in an individual. In some embodiments, SD refers to insufficient shrinkage of target lesions to qualify for PR or CR (such as those described above) and insufficient increase in target lesion(s) to qualify for progressive disease (PD). In some embodiments, PD refers to at least a 20% increase in the sum of the diameters of all target lesions measured, and the sum shows an absolute increase of at least 5 mm, based on the minimum sum of the diameters of all target lesions recorded at or after baseline, and the sum shows an absolute value.

[0068] In some embodiments, the methods of the present disclosure are used to treat inoperable, locally advanced, metastatic, and / or recurrent cancer (e.g., solid tumors).

[0069] In some embodiments, locally advanced or progressive cancer refers to cancer that only spreads to nearby tissue or lymph nodes.For example, in the case of locally advanced breast cancer, cancer may invade the skin of the breast or the lower muscle of the chest, or may invade multiple local lymph nodes (located in the calf or the soft tissue above and below the clavicle).A second example is lung cancer, and locally advanced lung cancer is when cancer grows in the airways, chest wall, or the membrane that surrounds the lungs (pleura).

[0070] In some embodiments, metastatic cancer refers to cancer that has spread to a distant part or parts of the body. For example, the most common places for metastatic breast cancer to spread are the bones, lungs, brain, and liver. A second example is metastatic lung cancer, where the cancer has spread to the brain, bones, liver, adrenal glands, other parts of the lungs, or other lungs.

[0071] In some embodiments, the cancer (e.g., a solid tumor) treated by the methods of the present disclosure is histologically and / or cytologically proven.

[0072] In some embodiments, in the immune modulatory oligonucleotide-antibody conjugates herein, the oligonucleotide and the antibody are linked together via a linking moiety. In some embodiments, one antibody can be conjugated to one oligonucleotide (DAR1), as shown in FIG. 1. In some embodiments, the oligonucleotide-antibody conjugate is a conjugate comprising an antibody or antigen-binding fragment thereof and one or more immune modulatory oligonucleotides (P), wherein the antibody or antigen-binding fragment is linked to one or more Q tag peptides (Q) comprising at least one glutamine residue, each immune modulatory oligonucleotide being linked via an amide bond with the glutamine residue of the Q tag peptide and a nucleotide sequence represented by the formula (A): [ka] or a stereoisomer, a mixture of two or more diastereomers, a tautomer, or a mixture of two or more tautomers thereof, or a pharma- ceutically acceptable salt, solvate, or hydrate thereof; During the ceremony, [ka] indicates the attachment point of each Q to an antibody or antigen-binding fragment (Ab), each Q is independently a Q tag peptide sequence containing at least one glutamine residue; each L is independently a bond or a linker moiety connected to Q via an amide bond with a glutamine residue; Each P is independently an immune modulatory oligonucleotide or a pharma- ceutically acceptable salt thereof.

[0073] In another aspect, the oligonucleotide is conjugated to the polypeptide via a linking moiety, the length, rigidity, and chemical composition of which affect the kinetics of conjugation and the stability of the resulting conjugate.

[0074] Linker portion (L) In some embodiments, the linking moiety comprises polyethylene glycol (PEG). In some embodiments, the PEG contains about 10-50 ethylene glycol units. In some embodiments, the linking moiety is an aliphatic chain.

[0075] In formula (A), the linking moiety is represented by L. In some embodiments, the linker L comprises an oligoethylene glycol or polyethylene glycol moiety. In certain embodiments, the linker L has the structure [ka] is a group having the formula [ka] indicates the point of attachment to the oligonucleotide P, [ka] indicates the attachment point of the Q tag to a glutamine residue.

[0076] In other embodiments, the linker L has the structure [ka] is a group having the formula [ka] indicates the point of attachment to the oligonucleotide P, [ka] indicates the point of attachment of the Q tag to a glutamine residue. 1 is absent. In some embodiments, L 1 is unsubstituted alkyl. In some embodiments, L 1 are independently unsubstituted C 1~6 In some embodiments, each L 1 is methyl or ethyl. In some embodiments, L 1 is independently substituted alkyl. In some embodiments, L 1 are independently unsubstituted C 1~6 In some embodiments, L 1 is a C substituted with one or more substituents selected from the group consisting of alkoxy, acyl, acyloxy, alkoxycarbonyl, carbonylalkoxy, acylamino, amino, aminoacyl, aminocarbonylamino, aminocarbonyloxy, cycloalkyl, cycloalkenyl, cyano, azido, halo, hydroxyl, nitro, carboxyl, thiol, thioalkyl, alkyl, alkenyl, alkynyl, heterocyclyl, aminosulfonyl, sulfonylamino, sulfonyl, and oxo. 1~6 It is an alkyl.

[0077] In some embodiments, L 2 is absent. In some embodiments, L 2 is unsubstituted or substituted alkyl.

[0078] In some embodiments, L 3 is absent. In some embodiments, L 3 is a linker moiety. In some embodiments, the linker moiety is unsubstituted or substituted alkyl. In some embodiments, the linker moiety is independently unsubstituted C 1~6 In some embodiments, the linker moieties are independently substituted alkyl. In some embodiments, the linker moieties are independently substituted C 1~6 In some embodiments, the linker moiety is a C substituted with one or more substituents selected from the group consisting of alkoxy, acyl, acyloxy, alkoxycarbonyl, carbonylalkoxy, acylamino, amino, aminoacyl, aminocarbonylamino, aminocarbonyloxy, cycloalkyl, cycloalkenyl, cyano, azido, halo, hydroxyl, nitro, carboxyl, thiol, thioalkyl, alkyl, alkenyl, alkynyl, heterocyclyl, aminosulfonyl, sulfonylamino, sulfonyl, and oxo. 1~6 In some embodiments, the linker moiety is an amino acid residue. In some embodiments, the amino acid is selected from the group consisting of glycine, alanine, glutamic acid, and proline. In some embodiments, the linker is methyl. In some embodiments, the linker moiety is -R 5 C(O)R 6 NHR 7 -, wherein R 5 and R 7 is independently absent or unsubstituted or substituted alkyl; R 6is an amino acid residue. In some embodiments, the amino acid is selected from the group consisting of glycine, alanine, glutamic acid, and proline. In some embodiments, the linker moiety is -R 3 C(O)NHR 4 -, wherein R 3 and R 4 is independently absent or unsubstituted or substituted alkyl. In some embodiments, R 3 is methylene, R 4 is -(CH 2 ) 4 In some embodiments, R 3 is methylene, R 4 In the case of multiple oligonucleotides (i.e., p=2), two L 1 can be different or the same, and the two L 2 can be different or the same, and the two L 3 can be different or the same.

[0079] In some embodiments, m is about 3 to 10, about 10 to 15, about 15 to 20, about 20 to 25, about 25 to 30, about 5 to 16, about 15 to 30, about 15 to 25, or about 20 to 30. In some embodiments, m is 20, 21, 22, 23, 24, or 25.

[0080] Immunomodulatory Oligonucleotides In some embodiments, the immune modulatory oligonucleotide has the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, [ka] indicates the attachment point within the oligonucleotide, Each T 1 are independently O or S; Each T 2 O - or S- and T 3 The basis [ka] where: [ka] indicates the point of attachment to L, [ka] indicates the point of attachment to the remainder of the oligonucleotide, Z is O or S; U 5’ is -H or a halogen, R 5’ is -H or methoxy, R c1 is -H or methoxy, R g1 , R g2 , R g3 , and R g4 is H or oxo, R 3’ is methoxy, R 1 is C 1~4 -alkylene-hydroxy, R 2 is -H or methyl, n is an integer from 0 to 2.

[0081] In other embodiments, the immune modulatory oligonucleotide has the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, [ka] indicates the attachment point within the oligonucleotide, Each T 1 are independently O or S; Each T 2 O - or S - and T 3 The basis [ka] where: [ka] indicates the point of attachment to L, [ka] indicates the point of attachment to the remainder of the oligonucleotide, Z is O or S; R 5’ is -H or methoxy, R c1 is -H or methoxy, R g1 , R g2 , R g3 , and R g4 is H or oxo, R 3’ is methoxy, R 1 is C 1~4 -alkylene-hydroxy, R 2 is -H or methyl, n is an integer from 0 to 2.

[0082] In yet other embodiments, the immune modulatory oligonucleotide has the structure: [ka] or a pharma- ceutically acceptable salt thereof, During the ceremony, [ka] indicates the attachment point within the oligonucleotide, Each T 1are independently O or S; Each T 2 O - or S - and T 3 The basis [ka] where: [ka] indicates the point of attachment to L, [ka] indicates the point of attachment to the remainder of the oligonucleotide, Z is O or S; R 5’ is -H or methoxy, R c1 is -H or methoxy, R g1 , R g2 , R g3 , and R g4 is H or oxo, R 3’ is methoxy, R 1 is C 1~4 -alkylene-hydroxy, R 2 is -H or methyl, n is an integer from 0 to 2.

[0083] In some embodiments, the immune modulatory oligonucleotide comprises an oligonucleotide sequence shown in Table A, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide is an oligonucleotide shown in Table A, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide comprises an oligonucleotide shown in Table A, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) plus linker comprises an oligonucleotide selected from the group consisting of SEQ ID NOs: 34-35, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) plus linker comprises an oligonucleotide of SEQ ID NO: 34, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) plus linker comprises an oligonucleotide of SEQ ID NO: 35, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) is an oligonucleotide selected from the group consisting of SEQ ID NOs: 14-25, 141-152, and 162, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) comprises an oligonucleotide selected from the group consisting of SEQ ID NOs: 14-25, 141-152, and 162, or a pharma- ceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (P) comprises an oligonucleotide selected from the group consisting of SEQ ID NOs: 14-25, 141-152, and 162, or a pharma- ceutically acceptable salt thereof. Table A. Modified Oligonucleotides [Table 1-1] [Table 1-2] [Table 1-3] [Table 2]

[0084] In some embodiments, the immune modulatory oligonucleotide (e.g., CpG oligonucleotide) comprises an oligonucleotide sequence shown in Table A (compound number ending in "a"), or a pharmaceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (e.g., CpG oligonucleotide) is an oligonucleotide selected from the group consisting of the oligonucleotides in Table 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (e.g., CpG oligonucleotide) comprises an oligonucleotide selected from the group consisting of the oligonucleotides in Table 9, or a pharmaceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (e.g., CpG oligonucleotide) is compound 7.6a or compound 7.7a, or a pharmaceutically acceptable salt thereof. In some embodiments, the immune modulatory oligonucleotide (e.g., CpG oligonucleotide) is compound 7.7a, or a pharmaceutically acceptable salt thereof. Table 9. Modified oligonucleotide structures (-PEG 2 NH 2 (including [Table 3-1] [Table 3-2]

[0085] In some embodiments, the oligonucleotide is functionalized with a chemical tag for attachment to a linking moiety. In some embodiments, the chemical tag is attached to an internucleoside linkage of the oligonucleotide. In some embodiments, the chemical tag is attached to a 5' internucleoside linkage. In some embodiments, the chemical tag is attached to a 3' internucleoside linkage. In some embodiments, each internucleoside linkage is a phosphorothioate linkage. In some embodiments, each internucleoside linkage is a phosphorodithioate linkage. In some embodiments, the chemical tag is closer to the 5' end of the oligonucleotide than the 3' end. In some embodiments, the chemical tag is attached to a nucleobase.

[0086] In some embodiments of Formula (A), the linker L has the structure [ka] where m is 24; [ka] indicates the point of attachment to the oligonucleotide P, [ka] indicates the point of attachment of the Q tag to a glutamine residue, and oligonucleotide P has the structure: [ka] having During the ceremony, [ka] indicates the attachment point within the oligonucleotide, Each T 1 are independently O or S; Each T 2 O - or S - and T 3 The basis [ka] where: [ka] indicates the point of attachment to L, [ka] indicates the point of attachment to the remainder of the oligonucleotide, Z is O or S; U 5’ is -H or a halogen, R 5’ is -H or methoxy, R c1 is -H or methoxy, R g1 , R g2 , R g3 , and R g4 is H or oxo, R 3’ is methoxy, R 1 is C 1~4 -alkylene-hydroxy, R 2 is -H or methyl, n is an integer from 0 to 2.

[0087] In some embodiments, the linker oligonucleotide LP comprises an oligonucleotide sequence shown in Table A, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP is a compound shown in Table 10, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP is compound 7.6b or compound 7.7b, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP is compound 7.6b, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP is compound 7.7b, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP comprises an oligonucleotide of SEQ ID NO: 34, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP comprises an oligonucleotide of SEQ ID NO: 35, or a pharma- ceutically acceptable salt thereof. In some embodiments, the linker oligonucleotide LP comprises an oligonucleotide selected from the group consisting of SEQ ID NOs: 3-13, 26-38, 130-140, 153-161, and 163-166, or a pharma- ceutically acceptable salt thereof. Table 10. Modified oligonucleotide structures (-PEG 2 NHCOPEG 24 NH 2 (including [Table 4-1] [Table 4-2]

[0088] Anti-CD22 antibodies containing Q tags In some embodiments, the oligonucleotides of the disclosure are conjugated to an anti-CD22 antibody. In some embodiments, the oligonucleotides are conjugated to the antibody via one or more Q tags. In some embodiments, the Q tag comprises a glutamine residue linked to the remainder of the conjugate. In still further embodiments of this aspect that may be combined with any of the preceding embodiments, each Q tag independently comprises or is a peptide sequence selected from the group consisting of SEQ ID NOs: 39-55. In some embodiments, each Q tag independently comprises or is a peptide sequence selected from the group consisting of the peptide sequences in Table 3. In other embodiments of this aspect, each Q tag independently comprises or is a peptide sequence selected from the group consisting of SEQ ID NOs: 40-55. In still other embodiments, each Q tag independently comprises or is a peptide sequence selected from the group consisting of SEQ ID NOs: 47-49. In some embodiments, the Q tag comprises LLQGG (SEQ ID NO: 172), GGGLLQGG (SEQ ID NO: 173), RPQGF (SEQ ID NO: 47), or RPQGFGPP (SEQ ID NO: 49). In some embodiments, the Q tag comprises the peptide sequence RPQGF (SEQ ID NO: 47). In certain embodiments, the Q tag comprising the peptide sequence RPQGF (SEQ ID NO: 47) is selected from the group consisting of RPQGF (SEQ ID NO: 47), RPQGFPP (SEQ ID NO: 48), and RPQGFGPP (SEQ ID NO: 49). In some embodiments, the Q tag comprises the peptide sequence RPQGFGPP (SEQ ID NO: 49).

[0089] In some embodiments, the Qtags comprise one or more sequences shown in Table 3. [Table 5]

[0090] Certain aspects of the present disclosure relate to CD22 and anti-CD22 antibodies. In some embodiments, CD22 refers to human CD22 and the antibody binds to human CD22. CD22 is also known as Siglec-2, and CD22 gene and polypeptide sequences (e.g., human gene and polypeptide sequences) are well known in the art. See, for example, NCBI gene ID number 933 and NCBI reference sequence accession number NP_001172028. Any of the anti-CD22 antibodies provided herein find use in the conjugates and methods of the present disclosure.

[0091] In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain that comprises one, two, or three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain that comprises three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain that comprises one, two, or three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain that comprises three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain that comprises one, two, or three CDRs of a single antibody shown in Table 4, and a VL domain that comprises one, two, or three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain that comprises three CDRs of a single antibody shown in Table 4, and a VL domain that comprises three CDRs of a single antibody shown in Table 4. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain comprising a CDR-H1 comprising the sequence of SEQ ID NO: 113, a CDR-H2 comprising the sequence of SEQ ID NO: 115, and a CDR-H3 comprising the sequence of SEQ ID NO: 116. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain comprising a CDR-H1 comprising the sequence of SEQ ID NO: 114, a CDR-H2 comprising the sequence of SEQ ID NO: 189, and a CDR-H3 comprising the sequence of SEQ ID NO: 116. In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain comprising a CDR-L1 comprising the sequence of SEQ ID NO: 117, a CDR-L2 comprising the sequence of SEQ ID NO: 119, and a CDR-L3 comprising the sequence of SEQ ID NO: 120. In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain comprising a CDR-L1 comprising the sequence of SEQ ID NO: 118, a CDR-L2 comprising the sequence of SEQ ID NO: 177, and a CDR-L3 comprising the sequence of SEQ ID NO: 120. In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain comprising a CDR-L1 comprising the sequence of SEQ ID NO: 117, a CDR-L2 comprising the sequence of SEQ ID NO: 119, and a CDR-L3 comprising the sequence of SEQ ID NO: 121.In some embodiments, an antibody or conjugate of the disclosure comprises a VL domain comprising a CDR-L1 comprising the sequence of SEQ ID NO: 117, a CDR-L2 comprising the sequence of SEQ ID NO: 119, and a CDR-L3 comprising the sequence of SEQ ID NO: 122. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain comprising a CDR-H1 comprising the sequence of SEQ ID NO: 113, a CDR-H2 comprising the sequence of SEQ ID NO: 115, and a CDR-H3 comprising the sequence of SEQ ID NO: 116, and a VL domain comprising a CDR-L1 comprising the sequence of SEQ ID NO: 117, a CDR-L2 comprising the sequence of SEQ ID NO: 119, and a CDR-L3 comprising the sequence of SEQ ID NO: 120. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain comprising CDR-H1 comprising the sequence of SEQ ID NO: 113, CDR-H2 comprising the sequence of SEQ ID NO: 115, and CDR-H3 comprising the sequence of SEQ ID NO: 116, and a VL domain comprising CDR-L1 comprising the sequence of SEQ ID NO: 117, CDR-L2 comprising the sequence of SEQ ID NO: 119, and CDR-L3 comprising the sequence of SEQ ID NO: 121. In some embodiments, an antibody or conjugate of the disclosure comprises a VH domain comprising CDR-H1 comprising the sequence of SEQ ID NO: 113, CDR-H2 comprising the sequence of SEQ ID NO: 115, and CDR-H3 comprising the sequence of SEQ ID NO: 116, and a VL domain comprising CDR-L1 comprising the sequence of SEQ ID NO: 117, CDR-L2 comprising the sequence of SEQ ID NO: 119, and CDR-L3 comprising the sequence of SEQ ID NO: 122. [Table 6-1] [Table 6-2]

[0092] In certain embodiments, the anti-CD22 antibody is an antibody comprising a VH and VL as shown in Table 5 below. [Table 7-1] [Table 7-2] [Table 7-3]

[0093] In some embodiments, the anti-CD22 antibody or conjugate comprises a VH domain as set forth in Table 6 and a VL domain as set forth in Table 7. For example, in some embodiments, the anti-CD22 antibody or conjugate comprises a VH domain comprising a sequence selected from the group consisting of SEQ ID NOs: 64-67 and a VL domain comprising a sequence selected from the group consisting of SEQ ID NOs: 68-91. In some embodiments, the anti-CD22 antibody or conjugate comprises a VH domain comprising a sequence of SEQ ID NO: 65 and a VL domain comprising a sequence of SEQ ID NO: 73 or 87. In some embodiments, the anti-CD22 antibody or conjugate comprises a VH domain comprising a sequence of SEQ ID NO: 65 and a VL domain comprising a sequence of SEQ ID NO: 73. In some embodiments, the anti-CD22 antibody or conjugate comprises a VH domain comprising a sequence of SEQ ID NO: 65 and a VL domain comprising a sequence of SEQ ID NO: 87. [Table 8] [Table 9-1] [Table 9-2] [Table 9-3]

[0094] In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising a VH domain set forth in Table 6 and a heavy chain constant domain sequence set forth in Table 8A. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising a VL domain set forth in Table 7 and a light chain constant domain sequence set forth in Table 8A. [Table 10-1]

Table 10-2

Table 10-3

Table 10-4

Table 10-5

Table 10-6

Table 10-7

Table 10-8

Table 10-9

[0095] In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain (including a Q-tag) as set forth in Table 8B and a light chain as set forth in Table 8B. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising the sequence of SEQ ID NO: 179 or 180 and a light chain comprising the sequence of SEQ ID NO: 181 or 182. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising the sequence of SEQ ID NO: 179 and a light chain comprising the sequence of SEQ ID NO: 181. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising the sequence of SEQ ID NO: 179 and a light chain comprising the sequence of SEQ ID NO: 182. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising the sequence of SEQ ID NO: 180 and a light chain comprising the sequence of SEQ ID NO: 181. In some embodiments, the anti-CD22 antibody or conjugate comprises a heavy chain comprising the sequence of SEQ ID NO: 180 and a light chain comprising the sequence of SEQ ID NO: 182. In some embodiments, the anti-CD22 antibody or conjugate comprises two heavy chains, each comprising the sequence of SEQ ID NO: 179 or 180, and two light chains, each comprising the sequence of SEQ ID NO: 181 or 182. In some embodiments, the anti-CD22 antibody or conjugate comprises two heavy chains, each comprising the sequence of SEQ ID NO: 179, and two light chains, each comprising the sequence of SEQ ID NO: 181. In some embodiments, the anti-CD22 antibody or conjugate comprises two heavy chains, each comprising the sequence of SEQ ID NO: 179, and two light chains, each comprising the sequence of SEQ ID NO: 182. In some embodiments, the anti-CD22 antibody or conjugate comprises two heavy chains, each comprising the sequence of SEQ ID NO: 180, and two light chains, each comprising the sequence of SEQ ID NO: 181. In some embodiments, the anti-CD22 antibody or conjugate comprises two heavy chains, each comprising the sequence of SEQ ID NO: 180, and two light chains, each comprising the sequence of SEQ ID NO: 182. [Table 11-1] [Table 11-2]

[0096] In some embodiments, a conjugate according to Table 11 (eg, conjugates A-F) is used in the methods of the disclosure. [Table 12]

[0097] III. Kit Also provided herein are kits that contain the conjugates described above for use in any of the methods described herein.

[0098] In another aspect, the kit further comprises a package insert containing, but not limited to, suitable instructions for preparation and administration of the formulation, side effects of the formulation, and any other relevant information. The instructions may be in any suitable format, including, but not limited to, printed matter, videotape, computer readable disk, optical disk, or instructions to internet-based instructions.

[0099] In another aspect, a kit for treating an individual suffering from or susceptible to a condition described herein is provided, the kit comprising a first container containing a dosage amount of a composition or formulation disclosed herein and a package insert for use. The container can be any of those known in the art and suitable for storing and delivering intravenous formulations. In certain embodiments, the kit further comprises a second container comprising a pharma- ceutically acceptable carrier, diluent, adjuvant, etc. for preparation of the formulation to be administered to the individual.

[0100] In another aspect, kits may be provided that contain a sufficient dosage of a composition described herein (including a pharmaceutical composition thereof) to provide effective treatment to an individual for an extended period of time, such as, for example, 1-3 days, 1-5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 1 cycle, 2 cycles, 3 cycles, 4 cycles, 5 cycles, 6 cycles, 7 cycles, 8 cycles or more.

[0101] In some embodiments, the kits may also include multiple doses, and may be packaged with pharmaceutical compositions and instructions, in amounts sufficient for storage and use in a pharmacy (e.g., hospital pharmacy and compounding pharmacy). In certain embodiments, the kits may include dosage amounts of at least one composition disclosed herein. EXAMPLES

[0102] The subject matter of the present disclosure will be better understood by reference to the following examples, which are provided as illustrations of the invention and not as limitations.

[0103] Example 1: An open-label, dose-escalation study to evaluate the safety, tolerability, pharmacokinetics, preliminary efficacy, and pharmacodynamics of an anti-CD22-CpG conjugate in patients with advanced or metastatic solid tumors The conjugate comprising an anti-CD22 antibody (Ab) and an immune modulatory oligonucleotide (also referred to as an "anti-CD22-CpG conjugate") used in this study is a Toll-like receptor (TLR) agonist antibody conjugate (TRAAC) designed to deliver TLR9 activity to target immune cells via systemic administration. The anti-CD22-CpG conjugate contains a TLR9 agonist oligonucleotide (14-mer) moiety specifically conjugated to an anti-CD22 antibody at the C-terminus via a Q-tag peptide as described herein. Preclinical mouse tumor models have shown that anti-CD22-CpG conjugate treatment activates the TLR9 pathway in B cells to increase immune activation, cytokine production, antigen presentation, and anti-tumor effects. This study was designed to test the safety, tolerability, pharmacokinetics, preliminary efficacy, and pharmacodynamics of anti-CD22-CpG conjugate treatment in patients with advanced or metastatic solid tumors.

[0104] In some embodiments, one of conjugates A-F was administered, as shown in Table 11.

[0105] Anti-CD22-CpG conjugate C was administered to patients every 2 weeks (Q2W) starting at 0.1 mg / kg by intravenous infusion over approximately 90 minutes. Patients were monitored for daily temperature and CRS symptoms for the first week after receiving the first dose. Patients were monitored for 28 days to ensure no dose-limiting toxicities (DLTs) occurred, with a target rate of 25%. All patients were also monitored for delayed DLTs on days 29-60. In the absence of DLTs or grade 2 or higher cytokine release syndrome (CRS), the next group of patients was titrated to the next dose of 0.3 mg / kg Q2W with 28 days of monitoring. This process was continued with approximately 3-fold increases per dose (i.e., dose 1=0.1 mg / kg, dose 2=0.3 mg / kg, dose 3=1.0 mg / kg, dose 4=3.0 mg / kg). Further escalations are dose 5 = 10.0 mg / kg, dose 6 = 30.0 mg / kg, and optionally dose 7 = up to 60.0 mg / kg. If DLT or grade 2 or higher CRS is observed, subsequent dose level escalations will be up to 2-fold (e.g., if DLT is observed at dose 1 = 0.1 mg / kg, then up to dose 2 = 0.2 mg / kg). Dose level 7 is optional, will not exceed 60 mg / kg, and will depend on available safety, pharmacokinetic, and efficacy data collected from other dose cohorts. A Bayesian optimal interval with 3+3 design run-in (BOIN) design was applied to inform dose escalation / deescalation decisions for dose levels. Up to 30 patients may be enrolled in the dose escalation phase to determine the maximum tolerated dose (MTD). At least two dose optimization cohorts will be enrolled at dose levels equal to or below the MTD to confirm the most ideal dose for further evaluation or to identify a recommended phase 2 dose (RP2D). A total of 100 patients may be enrolled in the dose escalation and dose optimization phase, with a maximum of 5 patients initially enrolled at a particular dose, starting with a 48-hour interval between the first and subsequent patients to evaluate any early safety concerns.

[0106] The study included a screening period, a treatment period, and a follow-up period. All patients completed screening for up to 28 days. During the treatment period, patients received anti-CD22-CpG conjugates every other week (Q2W) until treatment discontinuation criteria were met. Patients then entered a follow-up period, which continued for up to 2 years or until study discontinuation criteria were met.

[0107] Recruitment criteria: (a) Age (18 years or older); (b) gender (all); (c) Inoperable, locally advanced, metastatic, or recurrent histologically or cytologically proven solid tumors of the following cancers: (i) cutaneous melanoma, (ii) breast cancer; (iii) ovarian cancer, (iv) colorectal cancer; (v) non-small cell lung cancer (NSCLC), (vi) renal cell carcinoma (RCC); (vii) head and neck squamous cell carcinoma (HNSCC); (viii) Merkel cell carcinoma, (ix) hepatocellular carcinoma (HCC), (x) Cervical squamous cell carcinoma, (xi) urothelial carcinoma, (xii) endometrial cancer; (xiii) esophagogastric adenocarcinoma; and (xiv) bile duct carcinoma, (d) Patients must have at least one measurable lesion as defined by RECIST v1.1 (e) Adult male or female patients aged 18 years or older on the date of signing the informed consent. (f) Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1; (g) Demonstration of adequate organ function.

[0108] Exclusion criteria: (a) History of or ongoing malignancy other than that being treated in this study; (b) known brain metastases or extracranial disease; (c) receiving long-term systemic steroid therapy (>10 mg / day prednisone or equivalent) or any immunosuppressive therapy within 7 days prior to the first dose of study drug; (d) active autoimmune disease requiring systemic treatment within the past 2 years; (e) History of immune-mediated adverse events of grade 3 or higher considered drug-related from previous immunotherapy; (f) Infection with HIV-1 or HIV-2 (unless HIV is well controlled and viral load is undetectable); Hepatitis B infection (unless disease is undetectable); Hepatitis C infection (unless disease is well controlled and viral load is negligible); (g) ongoing infection requiring systemic therapy; (h) significant cardiovascular disease within the past 6 months; (i) Administration of any type of systemic anticancer therapy within 4 weeks of the first dose of study drug; (j) current or previous participation in an interventional clinical trial with an investigational compound or device within 4 weeks of first dose of study drug; (k) Radiotherapy for bone metastases within 2 weeks of the first dose of study drug, or any other radiotherapy within 4 weeks; (l) major surgery within 2 weeks of first administration of study drug; (m) any live vaccine within 4 weeks of the first dose of the study drug, or any other vaccine within 2 days of the first dose of the study drug; (n) previous treatment with a TLR9 agonist or anti-CD22 antibody; (o) known hypersensitivity to any component of the test therapy or its analogues; (p) any previous or current evidence of any medical condition, laboratory abnormality, or other condition that would place the subject at risk by participating in the study, confound the results of the study, or prevent the patient's participation for the entire duration of the study; and (q) Known mental or substance abuse disorder that would prevent cooperation with the requirements of the examination.

[0109] Primary endpoints include an analysis of the safety and tolerability of increasing dose levels of the anti-CD22-CpG conjugate to identify the dose recommended for phase 2 expansion. Endpoints include assessment of 28-day dose-limiting toxicities (DLTs), adverse events (AEs), and laboratory abnormalities. AEs will be characterized by type, frequency, and severity according to the National Cancer Institute Common Terminology Criteria for Adverse Events (NCICTCAE) v5.0, and laboratory abnormalities will be assessed by CTCAE v5.0.

[0110] Secondary endpoints included: (a) Serum concentration (C max ), the shortest time to peak plasma concentration (T max ), area under the curve (AUC), drug clearance (CL), and half-life (t 1 / 2 Characterization of the pharmacokinetics of single and multiple doses of anti-CD22-CpG conjugates, including pharmacokinetic parameters such as (b) evaluation of the immunogenicity of the anti-CD22-CpG conjugate and the incidence of anti-drug antibodies against the anti-CD22-CpG conjugate; and (c) Assessment of preliminary antitumor activity assessed by best overall response rate (ORR) by RECISTv1.1 and iRECIST, as well as duration of response (DOR) and clinical benefit rate for all cohorts, and for ovarian cancer patients by a combination of RECISTv1.1 and Gynecologic Cancer Intergroup (GCIG) CA-125 ORR and DOR criteria. Response criteria are shown in Tables E1-E2C below.

[0111] Study endpoints included: (a) characterization of other efficacy outcomes, such as progression-free survival (PFS) and overall survival (OS); (b) Characterization of the extent of target binding by anti-CD22-CpG conjugates via measurement of CD22 expression on B cells in peripheral blood; (c) exploring the effect of anti-CD22-CpG conjugates on systemic soluble immune factors, including pre- and post-dose quantification of soluble immune factors in serum; (d) evaluation of the pharmacodynamic effects of anti-CD22-CpG conjugates on immune cell markers, including levels of phenotypic markers on B cells and lymphocyte counts in peripheral blood; and (e) exploring the effect of anti-CD22-CpG conjugates on pharmacodynamic markers in tumor tissue, including (i) tumor mutation burden (TMB), and (ii) pre- and post-dose levels of pharmacodynamic markers, including but not limited to PD-L1 and other immune regulatory markers. [Table 13] [Table 14] [Table 15-1] [Table 15-2] [Table 16] Best RECIST responses also require confirmation and maintenance for at least 28 days.

[0112] Example 2: CD22 target binding The ability of administered anti-CD22-CpG conjugates to demonstrate and maintain CD22 target (CD22) binding was evaluated. Using the protocol of Example 1, anti-CD22-CpG conjugate C was administered to patients as an intravenous infusion every other week (Q2W) at 0.1 mg / kg (1 patient), 0.3 mg / kg (1 patient), 1 mg / kg (7 patients), or 3 mg / kg (4 patients) over approximately 90 minutes. Peripheral whole blood (PB) was collected in cycle 1 before the first dose on day 1 (pre-C1D1), 3 hours after administration (C1D13 hours), then before administration of the second dose on day 2 (C1D2), day 8 (C1D8), day 15 (pre-C1D15), as well as at subsequent cycle time points.

[0113] PB-derived CD19 + B cells were analyzed for surface expression of CD22 by fluorescence-activated cell sorting (FACS) analysis using a Navios flow cytometer (Beckman Coulter) or a CytoFLEXLX cytometer (Beckman Coulter). Data analysis was performed using either WinList (Verity Software) or FlowLogic software. B cells were analyzed for surface expression of CD45 + CD19 + Lymphocytes were identified. Molecules of Equivalent Soluble Fluorochrome (MESF) values ​​of cell surface CD22 expression were determined using SpheroTech Rainbow Calibration Beads URCP-38-2K (SpheroTech) according to the manufacturer's recommendations. Data were compiled using GraphPad Prism software. Data were expressed as the mean percent change in CD22 cell surface expression from pre-dose values ​​+ / - standard deviation (SD).

[0114] As shown in Figure 2A, cell surface CD22 expression levels on peripheral B cells decreased after administration of anti-CD22-CpG conjugate. Higher doses tended to prolong target engagement as indicated by a slower rate of return of cell surface CD22 to pre-administration levels. Patients at 0.3 mg / kg and 1 mg / kg doses were re-evaluated after the second administration cycle. Figure 2B demonstrates that the anti-CD22-CpG conjugate at dose levels of 0.3 mg / kg and 1 mg / kg showed similar levels of target binding upon administration of the first and fourth doses. At the higher dose level of 3 mg / kg, the data show that the conjugate maintained target binding prior to and through the second treatment cycle (4 doses, 2 cycles administered every other week).

[0115] Example 3: B cell activation B cell activation in response to administration of anti-CD22-CpG conjugates was evaluated. B cells from PB collected from patients in Example 2 were analyzed for activation by measuring CD86 expression by FACS analysis using either a Navios flow cytometer (Beckman Coulter) or a CytoFLEXLX cytometer (Beckman Coulter). Data analysis was performed using either WinList (Verity Software) or FlowLogic software. Naive B cells were cloned using CD45 + CD19 + CD27 - and memory B cells were identified as CD45 + CD19 + CD27 + CD86 expression was derived by gating on the percentage of naive and memory B cells expressing CD86. Data were compiled using GraphPad Prism software and expressed as fold change in marker expression from pre-dosing.

[0116] As shown in Figures 3A and 3B, all doses of anti-CD22-CpG conjugate induced B cell activation as indicated by increased CD86 expression. Figure 3C provides evidence of B cell activation after the second dosing cycle (4 doses, 2 cycles of biweekly dosing).

[0117] Example 4: Pharmacokinetics The pharmacokinetics of anti-CD22-CpG conjugates was evaluated by measuring the concentration of the conjugate in patients receiving 0.1, 0.3, and 1 mg / kg of Example 2. A MesoScale Discoveries (MSD) assay using human CD22ECD (TNT9, Tallac Therapeutics) ligand binding format and electrochemiluminescence (ECL) technology was used to detect the conjugates. The conjugates were detected using biotin-conjugated antisense oligos (primary detection agent) and Sulfo-TAG-labeled streptavidin. The electrochemiluminescence signal after addition of Read Buffer T was read on an MSDSector Imager plate reader. The results are shown in Table E4 and Figure 4. [Table 17]

[0118] Conjugate C max The increase in AUC was approximately dose-proportional between the 0.1 and 1 mg / kg doses. The increase in AUC was greater than dose-proportional at the 0.1 and 1 mg / kg doses. A similar trend was observed when samples were assayed for total antibody.

[0119] At the time of filing, three of eight evaluable subjects had stable disease according to the criteria in Example 1. These subjects were at the 0.3, 1, and 3 mg / kg dose levels.

Claims

1. 1. A method of treating cancer in an individual, comprising administering to said individual a conjugate comprising an anti-CD22 antibody (Ab) and an immune modulatory oligonucleotide (P) at a dose of 0.1 mg / kg to 60 mg / kg; The Ab comprises two antibody light chains, two antibody heavy chains, and two Q tag peptides (Q), each of the two Q tag peptides comprising the amino acid sequence RPQGFGPP (SEQ ID NO:49), wherein one Q tag peptide is linked to the C-terminus of each of the two antibody heavy chains, and one of the two Q tag peptides is linked to the immune modulatory oligonucleotide via an amide bond with a glutamine residue of the Q tag peptide and a linker (L) shown in formula (A); 【Chemistry 1】 During the ceremony, 【Chemistry 2】 indicates the point of attachment of Q to the antibody (Ab), wherein each heavy chain of said Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116; and each light chain of said Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 120-122; wherein the linker L is 【Chemistry 3】 and wherein m is 24; 【Chemistry 4】 indicates the point of attachment to the oligonucleotide P, 【Chemistry 5】 indicates the point of attachment of the Q tag to the glutamine residue, wherein said oligonucleotide P has the structure: 【Chemistry 6】 or 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof, During the ceremony, 【Chemistry 8】 indicates a point of attachment within the oligonucleotide, 【Chemistry 9】 indicates the point of attachment to the linker L.

2. 10. The method of claim 1, wherein the individual has an inoperable, locally advanced, metastatic, and / or recurrent solid tumor.

3. 3. The method of claim 1 or 2, wherein the individual has a cancer selected from the group consisting of cutaneous melanoma, breast cancer, ovarian cancer, colorectal cancer, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), head and neck squamous cell carcinoma (HNSCC), Merkel cell carcinoma, hepatocellular carcinoma (HCC), cervical squamous cell carcinoma, urothelial carcinoma, endometrial carcinoma, gastroesophageal adenocarcinoma, and cholangiocarcinoma.

4. 1. A method of treating cancer in an individual, comprising administering to the individual a conjugate comprising an anti-CD22 antibody (Ab) and an immune modulatory oligonucleotide (P), wherein the individual has an inoperable, locally advanced, metastatic, and / or recurrent solid tumor; The Ab comprises two antibody light chains, two antibody heavy chains, and two Q tag peptides (Q), each of the two Q tag peptides comprising the amino acid sequence RPQGFGPP (SEQ ID NO:49), wherein one Q tag peptide is linked to the C-terminus of each of the two antibody heavy chains, and one of the two Q tag peptides is linked to the immune modulatory oligonucleotide via an amide bond with a glutamine residue of the Q tag peptide and a linker (L) shown in formula (A); 【Chemistry 10】 During the ceremony, 【Chemistry 11】 indicates the point of attachment of Q to the antibody (Ab), wherein each heavy chain of said Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116; and each light chain of said Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 120-122; wherein the linker L is 【Chemistry 12】 and wherein m is 24; 【Chemistry 13】 indicates the point of attachment to the oligonucleotide P, 【Chemistry 14】 indicates the point of attachment of the Q tag to the glutamine residue, wherein said oligonucleotide P has the structure: 【Chemistry 15】 or 【Chemistry 16】 or a pharma- ceutically acceptable salt thereof, During the ceremony, 【Chemistry 17】 indicates a point of attachment within the oligonucleotide, 【Chemistry 18】 indicates the point of attachment to the linker L.

5. 5. The method of claim 4, wherein the individual has a cancer selected from the group consisting of cutaneous melanoma, breast cancer, ovarian cancer, colorectal cancer, non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), head and neck squamous cell carcinoma (HNSCC), Merkel cell carcinoma, hepatocellular carcinoma (HCC), cervical squamous cell carcinoma, urothelial carcinoma, endometrial carcinoma, gastroesophageal adenocarcinoma, and cholangiocarcinoma.

6. 6. The method of claim 4 or claim 5, wherein the conjugate is administered to the individual at a dose of 0.1 mg / kg to 60 mg / kg.

7. The method of any one of claims 1 to 6, wherein the individual has a solid tumor that has progressed on and / or is intolerant to standard treatment.

8. 8. The method of any one of claims 1 to 7, wherein the conjugate is administered to the individual at a dose of 0.1, 0.3, 1.0, 3.0, 10.0, 30.0, or 60.0 mg / kg.

9. The method of any one of claims 1 to 8, wherein the conjugate is administered to the subject once every two weeks.

10. The method of any one of claims 1 to 9, wherein each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO:

120.

11. The method of any one of claims 1 to 9, wherein each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO:

121.

12. The method of any one of claims 1 to 9, wherein each heavy chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 113, a CDR2 comprising the amino acid sequence of SEQ ID NO: 115, and a CDR3 comprising the amino acid sequence of SEQ ID NO: 116, and each light chain of the Ab comprises a CDR1 comprising the amino acid sequence of SEQ ID NO: 117, a CDR2 comprising the amino acid sequence of SEQ ID NO: 119, and a CDR3 comprising the amino acid sequence of SEQ ID NO:

122.

13. The method of any one of claims 1 to 9, wherein each heavy chain of the Ab comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:

65.

14. The method of any one of claims 1 to 9 and 13, wherein each light chain of the Ab comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:

73.

15. The method of any one of claims 1 to 9 and 13, wherein each light chain of the Ab comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:

87.

16. The method of any one of claims 1 to 9, wherein each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179 or 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO: 181 or 182.

17. 17. The method of claim 16, wherein each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO:

181.

18. 17. The method of claim 16, wherein each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 179, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO:

182.

19. The method of claim 16, wherein each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO:

181.

20. The method of claim 16, wherein each heavy chain of the Ab comprising the Q tag peptide comprises the amino acid sequence of SEQ ID NO: 180, and each light chain of the Ab comprises the amino acid sequence of SEQ ID NO:

182.

21. The method of any one of claims 1 to 20, wherein the individual is a human.

22. The conjugate is (a) the conjugate at a concentration of 30 mg / mL; (b) 20 mM citrate; (c) 150 mM L-arginine; (d) 50 mM NaCl, and (e) 0.02% (w / v) polysorbate 80; The method of any one of claims 1 to 21, wherein the formulation has a pH of 6.

5.

23. The method of any one of claims 1 to 22, wherein the conjugate is administered to the individual intravenously.

Images