Antibody drug conjugates, combinations and methods of use including STING agonists

Abstract

The present disclosure provides a combination comprising a HER2-targeted STING agonist antibody-drug conjugate and a HER2-targeted therapeutic or immunotherapeutic agent. The present disclosure also provides the use of the combination in treatment, for example, the treatment of cancer. TIFF2025509266000018.tif58170

Description

[Technical field]

[0001] Related Applications This application claims priority to and benefit of U.S. Provisional Application No. 63 / 317,472, filed March 7, 2022, and U.S. Provisional Application No. 63 / 329,680, filed April 11, 2022, the contents of each of which are incorporated herein by reference in their entirety.

[0002] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (MRSN_037_001WO_SeqList_ST26.xml; size: 18,657 bytes; and creation date: March 1, 2023) are incorporated herein by reference in their entirety. [Background technology]

[0003] background Stimulator of interferon genes (STING) is a receptor in the endoplasmic reticulum that conveys innate immune sensing of cytosolic pathogen-derived DNA and self-DNA. STING is a 378 amino acid protein that mainly contains three structural domains: (i) an N-terminal transmembrane domain (aa1-154); (ii) a central globular domain (aa155-341); and (iii) a C-terminal tail (aa342-379). STING can form symmetric dimers with its ligands that combine in a V-shaped conformation but do not completely cover the bound ligand. STING agonists can bind to the pocket region of STING. However, the STING activation process is easily inhibited in some severe disease conditions, resulting in the inactivation of the STING pathway. Therefore, screening and designing potent STING agonists is of great importance for cancer immunotherapy and the treatment of other infectious diseases, including but not limited to obesity, liver injury, glucose-lipid metabolism, and viral infections. Specific targeting of immune pathways presents an opportunity for cancer therapy, potentially conferring greater specificity than cell population-based therapeutic approaches.

[0004] Antibody-drug conjugates (ADCs) are composed of drug-like small molecules covalently linked to antibodies. Antibodies represent a targeting mechanism that is tailored to a specific site of action. ADCs are designed to release small drug molecules when they reach the site, allowing ADCs to perform their designed function in a targeted manner, as opposed to diffusing systemically throughout the subject's body. This targeted approach allows treatment with drugs that would otherwise require high doses that would be toxic when administered systemically, and minimizes the possibility of on-target, off-tumor toxicity.

[0005] A key hallmark of the innate immune system is the recognition and elimination of foreign substances. Identification of these pathogenic invaders occurs through host recognition of evolutionarily conserved microbial structures known as pathogen-associated molecular patterns (PAMPs). Host recognition may occur through multiple pathways, including activation of pattern recognition receptors (PRRs), which ultimately lead to downstream signaling events resulting in an immune response.

[0006] The antibody-drug conjugate of the present disclosure can modulate the activity of STING, and thus provide beneficial therapeutic impact in the treatment of diseases, disorders, and / or conditions in which STING (stimulator of interferon genes) modulation is beneficial, including but not limited to inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, and as a vaccine adjuvant.In addition, combination therapy in which two or more drugs are used in a certain dosage regimen or administration form can increase efficacy by taking advantage of the additive or synergistic effects in the biological activities of two or more drugs.There is still a need for new immunotherapies for the treatment of diseases, especially cancer. Summary of the Invention

[0007] overview In some aspects, the present disclosure provides, inter alia, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapy or at least one immunotherapy (e.g., an immune checkpoint inhibitor), where the conjugate comprises an antibody or antigen-binding fragment thereof that specifically binds to an epitope of the human HER2 receptor.

[0008] In some aspects, the HER2-targeting STING agonist antibody-drug conjugate has the formula (A): It is a conjugate of TIFF2025509266000002.tif74151, wherein the conjugate is Variable heavy chain complementarity determining region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); variable heavy chain complementarity determining region 2 (CDRH2) comprising TIFF2025509266000003.tif4128; variable heavy chain complementarity determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 7); and Amino acid sequence variable light chain complementarity determining region 1 (CDRL1) comprising TIFF2025509266000004.tif4128; variable light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 13); and variable light chain complementarity determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 14). and d 15 is about 8.

[0009] In some aspects, the HER2 antibody or antigen-binding fragment thereof that specifically binds to an epitope of the human HER2 receptor includes residues 452-531 of the extracellular domain of the human HER2 receptor, residues 474-553 of SEQ ID NO:1, or residues 452-531 of SEQ ID NO:16.

[0010] In some aspects, the present disclosure provides compositions comprising combination therapies comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immunotherapeutic agent (e.g., an immune checkpoint inhibitor).

[0011] In some aspects, the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of HER2-targeted therapeutic or immunotherapeutic agents (e.g., immune checkpoint inhibitors).

[0012] In some aspects, the present disclosure provides compositions comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent.

[0013] In some aspects, the present disclosure provides compositions comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one immunotherapeutic agent (e.g., an immune checkpoint inhibitor).

[0014] In some aspects, the HER2 targeted therapeutic agent is an antibody or antigen-binding fragment thereof that specifically binds to HER2, a HER2 targeted antibody-drug conjugate that specifically binds to HER2, or a small molecule inhibitor of HER2.

[0015] In some aspects, the present disclosure provides compositions comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one antibody or antigen-binding fragment thereof that specifically binds to HER2, at least one HER2-targeted STING agonist antibody-drug conjugate that specifically binds to HER2, or at least one small molecule inhibitor of HER2.

[0016] In some aspects, the HER2 targeted therapeutic agent is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody.

[0017] In some aspects, the HER2 targeted therapy is trastuzumab, pertuzumab, a combination thereof, or margetuximab or a biosimilar thereof.

[0018] In some aspects, the HER2 targeted therapy is trastuzumab, pertuzumab, or a combination thereof.

[0019] In some aspects, the HER2 targeted therapy is margetuximab or a biosimilar thereof.

[0020] In some aspects, the HER2 targeted therapeutic agent is a HER2 targeted antibody-drug conjugate that specifically binds to HER2, such as ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®).

[0021] In some aspects, the combination includes at least one HER2-targeting STING agonist antibody-drug conjugate in combination with ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®).

[0022] In some aspects, the HER2 targeted therapeutic agent is a small molecule inhibitor of HER2, such as, for example, tucatinib, neratinib, or lapatinib.

[0023] In some aspects, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with tucatinib, neratinib, or lapatinib.

[0024] In some aspects, a combination comprising a HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one antibody or antigen-binding fragment thereof that specifically binds to HER2, at least one HER2-targeted STING agonist antibody-drug conjugate that specifically binds to HER2, or at least one small molecule inhibitor of HER2.

[0025] In some aspects, immune checkpoint inhibitors suitable for the combinations and methods of the present disclosure are monoclonal antibodies, humanized antibodies, fully human antibodies, fusion proteins, or combinations thereof.

[0026] In some aspects, immune checkpoint inhibitors suitable for the combinations and methods of the disclosure are PD-1 inhibitors or PD-L1 inhibitors.

[0027] In some aspects, the combination comprises a HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with a PD-1 inhibitor or a PD-L1 inhibitor.

[0028] In some aspects, the combination comprises a HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with an immune checkpoint inhibitor, such as avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab. In some aspects, the combination comprises a HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with dostallimab. In some aspects, the combination comprises a HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with pembrolizumab.

[0029] In some aspects, the combination comprising the HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with an immune checkpoint inhibitor, such as avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab.In some aspects, the combination comprising the HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dostallimab.In some aspects, the combination comprising the HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with pembrolizumab.

[0030] The combination of the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapeutic agent or the immune checkpoint inhibitor is useful for treating a condition such as cancer in a subject. For example, the combination comprising the HER2-targeted STING agonist antibody-drug conjugate disclosed herein and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor is useful for treating, preventing, delaying the progression of, or otherwise improving the symptoms of cancer in a subject.

[0031] In some embodiments, the cancer is selected from the group consisting of, e.g., anal cancer, astrocytoma, leukemia, lymphoma, head and neck cancer, liver cancer, testicular cancer, cervical cancer, sarcoma, hemangioma, esophageal cancer, eye cancer, laryngeal cancer, mouth cancer, mesothelioma, skin cancer, myeloma, oral cancer, rectal cancer, colorectal cancer, throat cancer, bladder cancer, breast cancer, urothelial cancer, uterine cancer, ovarian cancer, prostate cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, pancreatic cancer, renal cancer, gastric cancer, and gastric esophagogastric junction cancer.

[0032] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating a breast cancer symptom, a gastric cancer symptom, a gastroesophageal junction cancer symptom, a non-small cell lung cancer (NSCLC) symptom, or a colorectal cancer symptom in a subject.

[0033] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer in a subject.

[0034] In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving symptoms of breast cancer in a subject.In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving symptoms of HER2+ breast cancer in a subject.In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving symptoms of HER2- breast cancer in a subject.In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving symptoms of metastatic HER2+ breast cancer in a subject. In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving the symptoms of metastatic HER2+ breast cancer in subjects who have previously undergone at least one line, at least two lines, at least three lines, or at least four lines of breast cancer therapy.In some aspects, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise improving the symptoms of metastatic HER2+ breast cancer in subjects who have previously undergone three or more lines of breast cancer therapy.

[0035] In some aspects, the combination comprising HER2 targeting STING agonist antibody-drug conjugate and at least one HER2 targeting therapeutic agent or at least one immune checkpoint inhibitor used in any of the aspects of the method and use provided herein can be administered at any stage of disease.For example, such combination therapy can be administered to patients suffering from any stage of cancer, from early stage to metastasis.

[0036] The combination therapy comprising the HER2-targeting STING agonist antibody-drug conjugate and at least one HER2-targeting therapeutic agent or at least one immune checkpoint inhibitor used in any of these methods and uses can be administered without another therapeutic agent or in combination with one or more chemotherapeutic agents or other agents.In some aspects, the additional agent is any of the toxins described herein.In some aspects, the additional agent is (1) an EGFR inhibitor (e.g., a tyrosine kinase inhibitor or a targeted anti-EGFR antibody), (2) a BRAF inhibitor, (3) an ALK inhibitor, (4) a hormone receptor inhibitor, (5) an mTOR inhibitor, (6) a VEGF inhibitor, or (7) a cancer vaccine. In some aspects, the additional agent is, for example, ado-trastuzumab emtansine (Kadcyla), lapatinib, anastrozole, letrozole, exemestane, everolimus, fulvestrant, tamoxifen, toremifene, megestrol acetate, fluoxymesterone, ethinyl estradiol, paclitaxel, capecitabine, gemcitabine, eribulin, vinorelbine, cyclophosphamide, carboplatin, docetaxel, albumin-bound paclitaxel, cisplatin, epirubicin, ixabepivir, ribavirin ... In some aspects, the additional agent is Kadcyla (ado-trastuzumab emtansine).

[0037] In some aspects, the combination comprising HER2 targeting STING agonist antibody-drug conjugate, HER2 targeting therapeutic agent or immune checkpoint inhibitor, and additional agent is formulated into a single therapeutic composition, and the components are administered simultaneously.Alternatively, HER2 targeting STING agonist antibody-drug conjugate, HER2 targeting therapeutic agent or immune checkpoint inhibitor, and if present, additional agent are separate from each other, for example, each can be formulated into separate therapeutic composition and administered simultaneously, or administered at different times during treatment regimen.For example, HER2 targeting STING agonist antibody-drug is administered before administration of HER2 targeting therapeutic agent or immune checkpoint inhibitor combination; HER2 targeting STING agonist antibody-drug is administered after administration of HER2 targeting therapeutic agent or immune checkpoint inhibitor combination. As described herein, the combination of HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor is administered in a single dose or in multiple doses.

[0038] Pharmaceutical compositions according to the present disclosure can include a suitable carrier. These pharmaceutical compositions can be included in a kit, such as, for example, a diagnostic kit.

[0039] 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. In this specification, the singular form includes the plural unless the context clearly dictates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference. References cited herein are not admitted to be prior art to the claimed invention. In case of conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are merely illustrative and are not intended to be limiting. In case of conflict between the chemical structure and the name of a compound disclosed herein, the chemical structure will prevail.

[0040] Other features and advantages of the present disclosure will become apparent from the following detailed description and claims. [Brief description of the drawings]

[0041] [Figure 1] 1 is a graph showing the anti-tumor efficacy of conjugate 1, trastuzumab, the combination of conjugate 2 with trastuzumab, and the combination of conjugate 1 with trastuzumab in SKOV3 tumor-bearing mice at various dose levels and dosing regimens. [Diagram 2] 1 is a graph showing the antitumor efficacy of conjugate 1; combination of conjugate 1 with trastuzumab; combination of conjugate 1 with pertuzumab; combination of conjugate 1 with trastuzumab and pertuzumab; combination of conjugate 2 with trastuzumab; combination of conjugate 2 with pertuzumab; and combination of conjugate 2 with trastuzumab and pertuzumab in JIMT-1 tumor-bearing mice at various dose levels and dosing regimens. [Diagram 3]1 is a graph showing the anti-tumor efficacy of conjugate 1; conjugate 2; trastuzumab; pertuzumab; a combination of trastuzumab and pertuzumab; a combination of conjugate 1 and trastuzumab; a combination of conjugate 1 and pertuzumab; and a combination of conjugate 1, trastuzumab, and pertuzumab in SNU-5 tumor-bearing mice at various dose levels and dosing regimens. [Figure 4] 1 is a graph showing the anti-tumor efficacy of Conjugate 1; Conjugate 2; Enhertu; and the combination of Conjugate 1 and Enhertu in JIMT-1 tumor-bearing mice at various dose levels and dosing regimens. [Diagram 5] 1 is a graph showing the anti-tumor efficacy of Conjugate 3; Conjugate 4; anti-PD-1 RMP1-14; the combination of Conjugate 3 and anti-PD-1 RMP1-14; and the combination of Conjugate 4 and anti-PD-1 RMP1-14 in EMT6-RHER2_MSA tumor-bearing mice at various dose levels and dosing regimens. [Figure 6] Figures 6A and 6B are graphs showing tumor volume in mice previously treated with conjugate 4, upon re-challenge with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. Figures 6C and 6D are graphs showing tumor volume in mice previously treated with conjugate 4 and anti-PD-1 RMP1-14, upon re-challenge with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. [Figure 7] 1 is a series of graphs depicting PD-L1 expression in mouse and human SKOV3 tumors treated with vehicle, control ADC, and conjugate 1 (HER2-targeted STING agonist ADC). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0042] Detailed Description The present disclosure provides novel HER2-targeted STING agonist antibody-drug conjugates, pharmaceutical compositions containing them, and various uses of the conjugates.

[0043] definition Chemical names given to intermediate compounds and / or compounds of the present disclosure described herein may refer to any one of the tautomeric representations of such compounds (in some cases, such aliases are given to the experimental ones). It should be understood that any reference to a named compound (an intermediate compound or compound of the present disclosure) or a structurally depicted compound (an intermediate compound or compound of the present disclosure) is intended to encompass all tautomeric forms of such compounds, including zwitterionic forms and any mixtures thereof.

[0044] It should be understood that the terms "in some aspects," "in some aspects of the present disclosure," and "in some aspects of the compounds of the present disclosure" may be used interchangeably where appropriate.

[0045] The terms "about," "approximately," or "approximately," when used in connection with a numerical value, are meant to include a collection or range of values. In some aspects, "about X" includes a range of values ​​that are ±25%, ±20%, ±15%, ±10%, ±5%, ±2%, ±1%, ±0.5%, ±0.2%, or ±0.1% of X, where X is a numerical value. In some aspects, the term "about" refers to a range of values ​​that are 5% more or less than the specified value. In some aspects, the term "about" refers to a range of values ​​that are 2% more or less than the specified value. In some aspects, the term "about" refers to a range of values ​​that are 1% more or less than the specified value.

[0046] The listing of a range of values ​​is intended to serve as a shorthand way of referring to each separate value in the range individually, unless otherwise specified herein, and each separate value is incorporated herein as if it were listed individually herein. Ranges used herein include the two ends of the range, unless otherwise specified. In some aspects, the phrases "x is an integer between 1 and 6" and "x is an integer between 1 and 6" both mean "x is 1, 2, 3, 4, 5, or 6", i.e., the terms "X to Y" and "range of X to Y" include X and Y and the integers therebetween.

[0047] As used herein, the term "HER2" (also known as ErbB-2, NEU, HER-2, and CD340) refers to human epidermal growth factor receptor 2 (SwissProt P04626) and includes any variants, isoforms, and species homologs of HER2 that are naturally expressed by cells, including tumor cells, or expressed on cells transfected with the HER2 gene. Species homologs include rhesus monkey HER2 (macaca mulatta; Genbank Accession No. GI:109114897). These terms are synonymous and may be used interchangeably.

[0048] As used herein, the term "HER2 antibody" or "anti-HER2 antibody" is an antibody that specifically binds to the antigen HER2.

[0049] The term "antibody" as used herein is used in the broadest sense and includes various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired antigen-binding activity. The numbering of antibody amino acids is according to the Kabat EU Index (see Kabat, EA, et al., Sequences of Protein of immunological interest, Fifth Edition, US Department of Health and Human Services, US Government Printing Office (1991)).

[0050] The term "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of the intact antibody and binds to the antigen to which the intact antibody binds. Examples of antibody fragments include, but are not limited to, Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments.

[0051] As used herein, the term "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks the binding of the reference antibody to its antigen by 50% or more in a competitive assay, and conversely, the reference antibody blocks the binding of the antibody to its antigen by 50% or more in a competitive assay. Exemplary competitive assays are provided herein.

[0052] The terms "compete with" or "cross-compete with" when used herein in the context of two or more antibodies, refer to two or more antibodies competing for binding to HER2, where an antibody "blocks" or "cross-blocks" the binding of one or more other antibodies to HER2 if the antibody competes with the other antibody by 25% or more, with 25%-74% representing "partial blocking," and 75%-400% representing "full blocking." Unless otherwise defined or contradicted by context, the terms "compete with," "cross-compete with," "block," or "cross-blocking," as used herein, are also intended to encompass such antibody pairs.

[0053] The term "epitope" refers to a specific site on an antigen molecule to which an antibody binds.

[0054] The term "independently," as used herein, means that when more than one substituent is selected from a number of possible substituents, the substituents can be the same or different.

[0055] The term "pharmacologically acceptable" as used herein refers to compounds, conjugates, materials, compositions, and / or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0056] The term "pharmaceutical composition" as used herein refers to a mixture, formulation, or solution containing at least one therapeutic agent that is administered to a subject, such as a mammal or human, to treat a particular disease or condition that the subject is affected by. The pharmaceutical combination of the present invention can be formulated into pharmaceutical compositions suitable for enteral or parenteral administration, such as sugar-coated tablets, tablets, capsules or suppositories, or ampoules. If not otherwise specified, they are prepared in a manner known per se, for example, by various conventional mixing, grinding, direct compression, granulation, sugar-coating, dissolving, lyophilization processes, or fabrication techniques readily apparent to those skilled in the art. It will be understood that the unit content of the combination partner contained in each dose of each dosage form does not necessarily constitute an effective amount per se, since the required effective amount can be reached by administering multiple dosage forms. Those skilled in the art can select one or more of the above-mentioned carriers according to the specific desired properties of the dosage form, by routine experimentation and without any undue burden. The amount of each carrier used can vary within the range conventional in the art. The pharmaceutical compositions provided herein may be in the form of a sterile injectable preparation, such as a sterile injectable aqueous suspension or oily suspension. This suspension may be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents as mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a 1,3-butane-diol solution, or may be prepared as a lyophilized powder. Acceptable vehicles and solvents that may be employed include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils may be conventionally employed as a solvent or suspending medium. For this purpose, any non-irritating fixed oil may be employed, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid may also be used in the preparation of injectables.

[0057] As used herein, the term "treating" or "treat" describes the management and care of a patient for the purpose of combating a disease, condition, or disorder, and includes administering a compound of the present disclosure, or a pharma- ceutically acceptable salt, polymorph, or solvate thereof, to alleviate the symptoms or complications of the disease, condition, or disorder, or to eliminate the disease, condition, or disorder. The term "treat" can also include the treatment of a cell in vitro or the treatment of an animal model.

[0058] As used herein, the terms "preventing," "prevent," or "protecting against" describe reducing or eliminating the onset of symptoms or complications of such a disease, condition, or disorder.

[0059] As used herein, the term "subject" includes humans and non-human animals, as well as cell lines, cell cultures, tissues, and organs. In some aspects, the subject is a mammal. The mammal can be, for example, a human or a suitable non-human mammal, such as a primate, mouse, rat, dog, cat, cow, horse, goat, camel, sheep, or pig. The subject can also be a bird or poultry. In some aspects, the subject is a human.

[0060] As used herein, the term "subject in need thereof" refers to a subject who has a disease or is at increased risk of developing a disease. A subject in need thereof may be a subject who has previously been diagnosed or identified as having a disease or disorder disclosed herein. A subject in need thereof may also be a subject suffering from a disease or disorder disclosed herein. Alternatively, a subject in need thereof may be a subject who is at increased risk of developing such a disease or disorder compared to the overall population (i.e., a subject who is predisposed to developing such a disorder compared to the overall population). A subject in need thereof may have a refractory or resistant disease or disorder disclosed herein (i.e., a disease or disorder disclosed herein that does not respond or has not yet responded to treatment). A subject may be resistant at the start of treatment or may become resistant during treatment. In some embodiments, a subject in need thereof has undergone and failed all known effective therapies for a disease or disorder disclosed herein. In some embodiments, a subject in need thereof has undergone at least one therapy in the past.

[0061] The term "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent, including a conjugate of the present disclosure, that elicits the biological or medical response in a tissue system, animal, or human that is sought by a researcher, veterinarian, physician, or other clinician, including alleviation or partial alleviation of the symptoms of the disease, syndrome, condition, or disorder being treated.

[0062] "Effective amount," as defined herein, is intended to mean an amount of a conjugate that is sufficient to effectively treat or prevent when administered to a patient in need of such treatment. The amount of a given conjugate that corresponds to such an amount may be determined by the potency (pICso), efficacy (EC) of a particular conjugate, etc. 50The duration of treatment and administration of the conjugate (the time between doses and the timing of doses, e.g., before / with / after a meal) will vary depending on factors such as the mammalian host's biological half-life, disease state and its severity, the nature of the patient requiring treatment (e.g., age, size, and weight), but can nevertheless be routinely determined by one of skill in the art. Similarly, the duration of treatment and administration of the conjugate (the time between doses and the timing of doses, e.g., before / with / after a meal) will vary according to the nature of the mammal requiring treatment (e.g., body weight), the particular conjugate and its characteristics (e.g., pharmacokinetic properties), the disease or disorder and its severity, and the specific composition and method being used, but can nevertheless be determined by one of skill in the art.

[0063] The term "composition" refers to a product containing therapeutically effective amounts of the specified ingredients, and any product that results directly or indirectly from the combination of the specified ingredients in the specified amounts.

[0064] As used herein, the term "pharmaceutically acceptable excipient" means an excipient that is generally safe, non-toxic, and not biologically or otherwise undesirable and is useful for preparing pharmaceutical compositions, and includes excipients that are acceptable for veterinary and human pharmaceutical use. As used in the specification and claims, "a pharmaceutically acceptable excipient" includes both one such excipient and more than one such excipient.

[0065] The term "STING agonist" as used herein refers to a compound or moiety capable of interacting with STING, for example by binding to STING and / or inducing downstream signaling (e.g., characterized by activation of molecules associated with STING function). This includes direct phosphorylation of STING, IRF3, and / or NF-kB, and may also include STAT6. In some aspects, activation of the STING pathway results in increased production of type 1 interferons (mainly IFN-a and IFN-b) and / or increased expression of interferon-stimulated genes.

[0066] The term "STING agonist drug moiety" as used herein refers to a moiety derived from a STING agonist and capable of interacting with STING. In some aspects, a STING agonist drug moiety is a moiety derived from a STING agonist to allow the moiety to be linked to the remainder of a conjugate of the present disclosure.

[0067] "Immunotherapeutic agent," as used herein, refers to an agent that activates or suppresses the immune system or a component of the immune system. Exemplary immunotherapeutic agents include, but are not limited to, monoclonal antibodies, immune checkpoint inhibitors, vaccines, cytokine therapies, adoptive cellular therapies, or immune system modulators.

[0068] As used herein, "immune checkpoint inhibitor" or "immune checkpoint inhibitor" or "immune checkpoint blocker" or "immune checkpoint modulator" refers to an agent that binds to and blocks the activity of an inhibitory immune checkpoint protein, thereby allowing the immune system to recognize tumor cells and allowing a sustained immunotherapy response. The inhibition can be steric or allosteric, competitive or non-competitive inhibition. When the immune checkpoint protein is an immune stimulatory protein, the immune checkpoint inhibitor acts to promote the activity of the immune stimulatory protein, for example, by binding to and activating the stimulatory immune checkpoint protein, or by inhibiting by interfering with it, such as by binding to or inactivating the inhibitor of the stimulatory immune checkpoint protein. An example of an immune checkpoint inhibitor is an anti-immune checkpoint protein antibody.

[0069] As used herein, "immune checkpoint" refers to an inhibitory pathway of the immune system that is responsible for maintaining self-tolerance and modulating the duration and magnitude of physiological immune responses in peripheral tissues to minimize secondary tissue damage. Immune checkpoints are regulated by immune checkpoint proteins.

[0070] As used herein, "immune checkpoint protein" refers to a protein, such as a receptor (e.g., CTLA4 or PD-1) or a ligand (e.g., PD-L1), that regulates or modulates the extent of an immune response. Immune checkpoint proteins can be inhibitory or stimulatory. In particular, immune checkpoint proteins are inhibitory to the activation of immune responses. Thus, inhibition of inhibitory immune checkpoint proteins acts to stimulate or activate immune responses, such as T cell activation and proliferation.

[0071] As used herein, "combination therapy" refers to a treatment in which two or more therapeutic agents are administered to a subject to treat a disease or disorder, such as at least two or at least three therapeutic agents. For purposes herein, combination therapy includes a HER2-targeted STING agonist antibody-drug conjugate and a HER2-targeted therapeutic agent or an immune checkpoint inhibitor therapy.

[0072] As used herein, "co-administration," "co-administer," or "co-administered" refers to administration of at least two different therapeutic agents close enough in time. Such administration can be in any order, including simultaneous administration and orders spaced in time from a few seconds up to several days apart. Such administration can also include more than one administration of one agent and / or administration of the other agent independently. Administration of the agents can be by the same or different routes.

[0073] The term "co-administration" as used herein in relation to administration of pharmaceuticals refers to administration of pharmaceuticals such that the individual pharmaceuticals are present in the subject at the same time. Co-administration can include administration of pharmaceuticals at different times (via the same route or alternative routes) in addition to administration of pharmaceuticals at the same time (via the same route or alternative routes).

[0074] As used herein, the terms "at least one" of an item or "one or more" of an item each include a single item selected from a list, and a mixture of two or more items selected from a list.

[0075] As used herein, the term "immune response" relates to any one or more of the following: specific immune response, non-specific immune response, both specific and non-specific responses, innate response, primary immune response, adaptive immunity, secondary immune response, memory immune response, immune cell activation, immune cell proliferation, immune cell differentiation, and cytokine expression.

[0076] The conjugates of the present disclosure are useful in methods for treating or ameliorating viral infections, diseases, syndromes, conditions, or disorders affected by agonism of STING, which methods comprise, consist of, and / or consist essentially of administering to a subject, including animals, mammals, and humans, in need of such treatment, amelioration, and / or prevention, a therapeutically effective amount of a conjugate of the present disclosure, or an enantiomer, diastereomer, solvate, or pharma- ceutically acceptable salt thereof.

[0077] The term "conjugate of the present disclosure" or "conjugate of the present disclosure", as used herein, refers to a conjugate as defined herein in any form, i.e., any tautomeric form, any isomeric form, any salt form or non-salt form (e.g., as a free acid or free base form, or as a salt, particularly a pharma- ceutically acceptable salt thereof), as well as any physical form thereof (e.g., non-solid forms (e.g., liquid or semi-solid forms), as well as solid forms (e.g., amorphous or crystalline forms, designated polymorphic forms, solvated forms including hydrate forms (e.g., monohydrate, dihydrate, and hemihydrate)), as well as mixtures of various forms.

[0078] Thus, included within the present disclosure are the conjugates disclosed herein in any salt or non-salt form, and in any physical form thereof, as well as mixtures of various forms. Although such are included within the present disclosure, it will be understood that the conjugates of the present disclosure in any salt or non-salt form, and in any physical form thereof, may have different levels of activity, different bioavailability, and different handling characteristics for formulation purposes.

[0079] It is understood that throughout the description where a composition is described as having, including, or comprising specified components, it is contemplated that the composition also consists essentially of or consists of the listed components. Similarly, where a method or process is described as having, including, or comprising specified process steps, the process also consists essentially of or consists of the listed processing steps. Furthermore, it should be understood that the order of steps or the order for performing certain actions is not important so long as the invention remains operable. Furthermore, two or more steps or actions may be carried out simultaneously.

[0080] All percentages and ratios used herein are by weight unless otherwise specified. Other features and advantages of the present disclosure are apparent from the various examples. The examples provided illustrate various components and methodologies useful in practicing the present disclosure. The examples do not limit the claimed disclosure. Based on the present disclosure, a person skilled in the art can identify and employ other components and methodologies useful in practicing the present disclosure.

[0081] HER2 antibody In some aspects, the HER2 antibody suitable for conjugation binds to human HER2 in soluble form or membrane-bound form (i.e., when expressed on the cell surface). In some aspects, the present disclosure provides a monoclonal antibody that binds to HER2 and is humanized or fully human. In some aspects, the present disclosure provides a monoclonal antibody that specifically binds to HER2. These antibodies are collectively referred to herein as "HER2" antibodies.

[0082] In some aspects, a HER2 antibody suitable for conjugation has an equilibrium dissociation constant (K d or K D In some aspects, the present disclosure provides monoclonal antibodies that bind to HER2 and are humanized or fully human. For example, the HER2 antibodies provided herein have a K in the range of about ≦1 nM to about 1 pM. d It presents.

[0083] In some aspects, the HER2 antibody disclosed herein functions to modulate, block, inhibit, reduce, antagonize, neutralize, or otherwise interfere with the functional activity of HER2.In some aspects, the functional activity of HER2 includes, for example, the modulation of PI3K-Akt pathway activity.In some aspects, the HER2 antibody completely or partially inhibits HER2 functional activity by partially or completely modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or otherwise interfering with PI3K-Akt pathway activity.PI3K-Akt pathway activity is evaluated using any art-recognized method for detecting PI3K-Akt pathway activity, including but not limited to detecting phosphorylated Akt level in the presence and absence of the antibody or antigen-binding fragment disclosed herein.

[0084] In some aspects, a HER2 antibody is considered to completely modulate, block, inhibit, reduce, antagonize, neutralize, or otherwise interfere with HER2 functional activity when the level of HER2 functional activity in the presence of the HER2 antibody is reduced by at least 80%, e.g., 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, compared to the level of HER2 functional activity in the absence of binding to a HER2 antibody described herein. In some aspects, a HER2 antibody is considered to partially modulate, block, inhibit, reduce, antagonize, neutralize, or otherwise interfere with HER2 functional activity when the level of HER2 activity in the presence of the HER2 antibody is reduced by less than 95%, e.g., 10%, 20%, 25%, 30%, 40%, 50%, 60%, 75%, 80%, 85%, or 90%, compared to the level of HER2 activity in the absence of binding to a HER2 antibody described herein.

[0085] In some aspects, exemplary HER2 antibodies disclosed herein include the XMT-1519 antibody, which exhibits specificity for human HER2 and has been shown to inhibit the functional activity of HER2 in vitro.

[0086] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein, including the HER2 monoclonal antibody XMT-1519, comprise a heavy chain (HC), a heavy chain variable region (VH), a light chain (LC), and a light chain variable region (VL) as shown in the amino acid sequences and corresponding nucleic acid sequences presented in Table I below. The variable heavy and variable light chain regions of each antibody are shaded in the amino acid sequences below. The complementarity determining regions (CDRs) of the heavy and light chains are underlined in the amino acid sequences presented below.

[0087] Table I: Sequence of HER2 human or humanized monoclonal antibody XMT-1519 TIFF2025509266000005.tif77169

[0088] The antibodies and antigen-binding fragments thereof disclosed herein specifically bind to an epitope on the full-length human HER2 receptor comprising the amino acid sequence of SEQ ID NO:1.

[0089] The antibodies and antigen-binding fragments thereof disclosed herein specifically bind to an epitope on the extracellular domain (ECD) of the human HER2 receptor comprising the amino acid sequence of SEQ ID NO:16.

[0090] In some embodiments, the antibody disclosed herein exhibits different HER2 binding properties from the antibody described in the art.In some embodiments, the antibody disclosed herein binds to different epitopes of HER2, in that they cross-block each other, but do not cross-block Trastuzumab, Pertuzumab, Fab37 or chA21 from binding to HER2.In addition, in contrast to known antibodies, the antibody disclosed herein can efficiently internalize into HER2-expressing cells without promoting cell proliferation.

[0091] In some embodiments, the antibodies disclosed herein are fully human monoclonal antibodies that bind to novel epitopes and / or have other properties that are favorable for therapeutic use. In some embodiments, exemplary properties include, but are not limited to, favorable binding characteristics for cancer cells expressing high or low levels of human HER2, specific binding to recombinant human and cynomolgus HER2, efficient internalization upon binding to HER2, high ability to kill cancer cells expressing high or low levels of HER2 when administered as an antibody drug conjugate (ADC), no substantial agonistic effect on the proliferation of HER2-expressing cancer cells, and / or providing effective antibody-dependent cellular cytotoxicity (ADCC)-mediated killing of HER2-expressing cells, and any combination of the aforementioned properties.

[0092] In some aspects, the antibodies disclosed herein comprise: an epitope of the human HER2 receptor comprising residues 452-531 of the extracellular domain of the human HER2 receptor, residues 474-553 of SEQ ID NO: 1, or residues 452-531 of SEQ ID NO: 16 Also included are antibodies or antigen-binding fragments thereof that specifically bind to.

[0093] In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that bind to at least a portion of the N-terminus of domain IV of the human HER2 receptor, but do not cross-compete with antibodies that bind to epitope 4D5 of the human HER2 receptor. In some embodiments, the antibodies or antigen-binding fragments thereof described herein do not cross-compete with trastuzumab for binding to the human HER2 receptor, since trastuzumab is known to bind to epitope 4D5 of the human HER2 receptor. As used herein, the term epitope 4D5 of the human HER2 receptor refers to amino acid residues 529-627 of the extracellular domain of the human HER2 receptor, residues 551-649 of SEQ ID NO: 1, or residues 529-627 of SEQ ID NO: 16. In some embodiments, the antibodies or antigen-binding fragments thereof also bind to at least one epitope on the cynomolgus monkey HER2 receptor.

[0094] In some aspects, the antibodies disclosed herein comprise: an epitope of the human HER2 receptor comprising residues 452-500 of the extracellular domain of the human HER2 receptor, residues 474-522 of SEQ ID NO: 1, or residues 452-500 of SEQ ID NO: 16 Also included are antibodies or antigen-binding fragments thereof that specifically bind to.

[0095] In some aspects, the antibodies disclosed herein comprise: An epitope of the human HER2 receptor, comprising at least one of amino acid residues selected from amino acid residues E521, L525, and R530 of the extracellular domain of the human HER2 receptor, e.g., residues 543, 547, and 552 of SEQ ID NO: 1, and residues 521, 525, and 530 of SEQ ID NO: 16. Also included are antibodies or antigen-binding fragments thereof that specifically bind to the extracellular domain of the human HER2 receptor, comprising at least two amino acid residues selected from amino acid residues E521, L525, and R530 of the extracellular domain of the human HER2 receptor. In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of the human HER2 receptor that comprises at least amino acid residues E521, L525, and R530 of the extracellular domain of the human HER2 receptor. In some embodiments, any or all of these antibodies or their antigen-binding fragments also bind to at least one epitope on the cynomolgus monkey HER2 receptor.

[0096] In some embodiments, the antibody disclosed herein also includes an antibody or antigen-binding fragment thereof that binds to at least a portion of domain III and at least a portion of the N-terminus of domain IV of human HER2 receptor, but does not cross-compete with Fab37 monoclonal antibody or an antibody that binds to epitope 4D5 of human HER2 receptor.In some embodiments, the antibody or antigen-binding fragment thereof described herein does not cross-compete with Fab37 monoclonal antibody and / or trastuzumab for binding to human HER2 receptor.In some embodiments, the antibody or antigen-binding fragment thereof also binds to at least one epitope on cynomolgus monkey HER2 receptor.

[0097] In some aspects, the antibodies disclosed herein comprise: an epitope of the human HER2 receptor comprising residues 520-531 of the extracellular domain of the human HER2 receptor, residues 542-553 of SEQ ID NO: 1, or residues 520-531 of SEQ ID NO: 16 Also included are antibodies or antigen-binding fragments thereof that specifically bind to.

[0098] In some aspects, the antibodies disclosed herein comprise: an epitope of the human HER2 receptor comprising at least one amino acid residue selected from residues C453, H456, H473, N476, R495, G496, H497, and W499 of the extracellular domain of the human HER2 receptor, e.g., residues 475, 478, 495, 498, 517, 518, 519, and 521 of SEQ ID NO: 1, or residues 453, 456, 473, 476, 495, 496, 497, and 499 of SEQ ID NO: 16; Also included are antibodies or antigen-binding fragments thereof that specifically bind to the extracellular domain of human HER2 receptor. In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of the extracellular domain of human HER2 receptor that comprises at least two amino acid residues, at least three amino acid residues, at least four amino acid residues, at least five amino acid residues, or at least six amino acid residues selected from amino acid residues C453, H456, H473, N476, R495, G496, H497, and W499 of the extracellular domain of human HER2 receptor. In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of the extracellular domain of human HER2 receptor that comprises at least amino acid residues C453, H456, H473, N476, R495, G496, H497, and W499 of the extracellular domain of human HER2 receptor. In some embodiments, any or all of these antibodies or antigen-binding fragments thereof also bind to at least one epitope on the cynomolgus monkey HER2 receptor.

[0099] In some aspects, the antibodies disclosed herein comprise: an epitope of the human HER2 receptor comprising at least one amino acid residue selected from residues C453, H473, N476, R495, H497, and W499 of the extracellular domain of the human HER2 receptor, e.g., residues 475, 495, 498, 517, 519, and 521 of SEQ ID NO: 1, or residues 453, 473, 476, 495, 497, and 499 of SEQ ID NO: 16; Also included are antibodies or antigen-binding fragments thereof that specifically bind to the extracellular domain of the human HER2 receptor. In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of the extracellular domain of the human HER2 receptor that comprises at least two amino acid residues, at least three amino acid residues, at least four amino acid residues, at least five amino acid residues, or at least six amino acid residues selected from amino acid residues C453, H473, N476, R495, H497, and W499 of the extracellular domain of the human HER2 receptor. In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of the extracellular domain of the human HER2 receptor that comprises at least amino acid residues C453, H473, N476, R495, H497, and W499 of the extracellular domain of the human HER2 receptor. In some embodiments, any or all of these antibodies or their antigen-binding fragments also bind to at least one epitope on the cynomolgus monkey HER2 receptor.

[0100] In some embodiments, these antibodies show specificity for human HER2, and they have been shown to modulate, e.g., block, inhibit, reduce, antagonize, neutralize, or otherwise interfere with, the PI3K-Akt pathway, which promotes cell survival by reducing the level of phosphorylated AKT. In some embodiments, these antibodies are internalized from the cell surface of HER2-expressing cells at the same or substantially similar rate as that of trastuzumab or its biosimilar. In some embodiments, these antibodies and antigen-binding fragments have an internalization rate of about 50% of the total surface bound at time 0 is internalized by 4 hours later.

[0101] In some embodiments, the antibodies disclosed herein comprise a heavy chain variable region having an amino acid sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to a sequence selected from SEQ ID NO:2, and a light chain variable region having an amino acid sequence at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to a sequence selected from SEQ ID NO:9.

[0102] In some embodiments, the antibodies disclosed herein comprise a heavy chain amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO:4, and a light chain amino acid sequence that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more identical to the amino acid sequence of SEQ ID NO:11.

[0103] In some aspects, the antibodies disclosed herein comprise a heavy chain variable region amino acid sequence of SEQ ID NO:2 and a light chain variable region amino acid sequence of SEQ ID NO:9.

[0104] In some aspects, the antibody disclosed herein comprises a heavy chain amino acid sequence of SEQ ID NO:4 and a light chain amino acid sequence of SEQ ID NO:11.

[0105] In some aspects, the antibodies disclosed herein comprise a CDRH1 amino acid sequence of SEQ ID NO:5, a CDRH2 amino acid sequence of SEQ ID NO:6, a CDRH3 amino acid sequence of SEQ ID NO:7, a CDRL1 amino acid sequence of SEQ ID NO:12, a CDRL2 amino acid sequence of SEQ ID NO:13, and a CDRL3 amino acid sequence of SEQ ID NO:14.

[0106] In some embodiments, the antibodies disclosed herein comprise one or more conservative amino acid substitutions in the variable domain sequence, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more conservative substitutions in the variable domain sequence. In some embodiments, these conservative amino acid substitutions are in the CDR regions, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more conservative substitutions are made cumulatively across all CDRs. In some embodiments, there may be up to 1, 2, 3, or 4 conservative amino acid substitutions in each CDR sequence, e.g., SEQ ID NOs: 5-7 and 12-14.

[0107] Those skilled in the art will recognize that it is possible to determine without undue experimentation whether a monoclonal antibody has the same specificity as monoclonal antibody XMT-1519 by determining whether the former prevents the latter from binding to natural binding partners or other molecules known to associate with HER2. In some embodiments, if the monoclonal antibody being tested competes with the monoclonal antibody disclosed herein as shown by reduced binding by the monoclonal antibody disclosed herein, the two monoclonal antibodies bind to the same or closely related epitopes.

[0108] In some embodiments, an alternative method for determining whether a monoclonal antibody has the specificity of the monoclonal antibodies disclosed herein is to preincubate the monoclonal antibodies disclosed herein with soluble HER2 (to which the monoclonal antibodies disclosed herein are normally reactive), and then add the monoclonal antibody being tested to determine whether the ability of the monoclonal antibody being tested to bind to HER2 is inhibited. If the monoclonal antibody being tested is inhibited, then in all likelihood it has the same or a functionally equivalent epitopic specificity as the monoclonal antibodies disclosed herein.

[0109] In some embodiments, screening of the monoclonal antibodies disclosed herein can also be accomplished, for example, by measuring HER2-mediated PI3K-Akt pathway activity and determining whether the test monoclonal antibody can modulate, block, inhibit, reduce, antagonize, neutralize, or otherwise interfere with PI3K-Akt pathway activity. In some embodiments, HER2 antibodies suitable for conjugation can be made and purified by well-known techniques, for example, WO 2015 / 195917 and PCT / US2018 / 019873, each of which is incorporated herein by reference in its entirety.

[0110] HER2-targeted STING agonist antibody conjugate The present invention relates to combination therapy involving an immunoconjugate comprising a HER2-targeting antibody conjugated to a STING agonist.

[0111] In some aspects, the present disclosure provides, inter alia, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immunotherapeutic agent (e.g., an immune checkpoint inhibitor), wherein the conjugate comprises an antibody or antigen-binding fragment thereof that specifically binds to an epitope of the human HER2 receptor.

[0112] In some embodiments, the HER2-targeting STING agonist antibody-drug conjugate has formula (A): It is a conjugate of TIFF2025509266000006.tif74150, wherein the conjugate is Variable heavy chain complementarity determining region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); variable heavy chain complementarity determining region 2 (CDRH2) comprising TIFF2025509266000007.tif4128; variable heavy chain complementarity determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 7); and Amino acid sequence variable light chain complementarity determining region 1 (CDRL1) comprising the amino acid sequence GASSRAT (SEQ ID NO: 13); variable light chain complementarity determining region 2 (CDRL2) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 14); and d 15 is about 8.

[0113] In some aspects, an epitope of the human HER2 receptor comprising residues 452-531 of the extracellular domain of the human HER2 receptor, residues 474-553 of SEQ ID NO: 1, or residues 452-531 of SEQ ID NO: 16 A HER2 antibody or antigen-binding fragment thereof that specifically binds to HER2.

[0114] In some embodiments, d 15 is 2, 4, 6, or 8. In some embodiments, d 15 is 6 or 8.

[0115] In some embodiments, d 15 is 8. In some embodiments, d 15 is 6.

[0116] HER2 targeted therapy Any suitable HER2 targeting therapeutic agent is contemplated herein for use in the combination and method of the present disclosure.In some embodiments, the HER2 targeting therapeutic agent is the antibody or its antigen-binding fragment that specifically binds to HER2, the second HER2 targeting STING agonist antibody-drug conjugate that specifically binds to HER2, or the small molecule inhibitor of HER2.

[0117] In some embodiments, the HER2 targeted therapeutic agent is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody.

[0118] In some embodiments, the HER2 targeting therapy is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody. In some embodiments, the HER2 antibody, the HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody is trastuzumab, pertuzumab, or margetuximab, or a biosimilar thereof. In some embodiments, the HER2 targeting therapy is trastuzumab or pertuzumab, or a combination thereof. In some embodiments, the HER2 targeting therapy is trastuzumab. In some embodiments, the HER2 targeting therapy is pertuzumab. In some embodiments, the HER2 targeting therapy is margetuximab or a biosimilar thereof. In some embodiments, the HER2 targeted therapy is a biosimilar of trastuzumab, a biosimilar of pertuzumab, or a combination of a biosimilar of trastuzumab and a biosimilar of pertuzumab. In some embodiments, the HER2 targeted therapy is a biosimilar of trastuzumab. In some embodiments, the HER2 targeted therapy is a biosimilar of pertuzumab. In some embodiments, the HER2 targeted therapy is a combination of a biosimilar of trastuzumab and a biosimilar of pertuzumab.

[0119] In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with at least one HER2 antibody, at least one HER2 dimerization inhibitor antibody, or a combination of at least one HER2 antibody and at least one HER2 dimerization inhibitor antibody.In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with trastuzumab or pertuzumab, or a combination thereof, or margetuximab.In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with trastuzumab or pertuzumab, or a combination thereof.In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with margetuximab. In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with at least one trastuzumab biosimilar or at least one pertuzumab biosimilar, or a combination thereof, or at least one margetuximab biosimilar. In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with at least one trastuzumab biosimilar or at least one pertuzumab biosimilar, or a combination thereof. In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate in combination with at least one margetuximab biosimilar.

[0120] In some embodiments, the combination comprising at least one of the present disclosure's HER2-targeting STING agonist antibody-drug conjugates is administered in combination with at least one HER2 antibody, at least one HER2 dimerization inhibitor antibody, or a combination thereof, or with margetuximab.In some embodiments, the combination comprising at least one of the present disclosure's HER2-targeting STING agonist antibody-drug conjugates is administered in combination with at least one trastuzumab or at least one pertuzumab, or a combination thereof, or with at least one margetuximab.In some embodiments, the combination comprising at least one of the present disclosure's HER2-targeting STING agonist antibody-drug conjugates is administered in combination with at least one trastuzumab biosimilar or at least one pertuzumab biosimilar, or a combination thereof, or with at least one margetuximab biosimilar.

[0121] In some embodiments, the HER2 targeted therapeutic agent is a HER2 targeted antibody-drug conjugate that specifically binds to HER2. In some embodiments, the HER2 targeted antibody-drug conjugate is, for example, ado-trastuzumab emtansine (T-DM1) (Kadcyla®) fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®), or a biosimilar thereof.

[0122] In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with at least one HER2-targeting antibody-drug conjugate. In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®). In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with a biosimilar of ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or a biosimilar of fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®). In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or a biosimilar thereof. In some embodiments, the combination comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) or a biosimilar thereof.

[0123] In some embodiments, a combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one HER2-targeting antibody-drug conjugate. In some embodiments, a combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®).

[0124] In some embodiments, the HER2 targeting therapeutic agent is a small molecule inhibitor of HER2.In some embodiments, the small molecule inhibitor of HER2 is, for example, tucatinib, neratinib, or lapatinib.In some embodiments, the combination comprises at least one of the HER2 targeting STING agonist antibody-drug conjugates of the present disclosure combined with at least one of the small molecule inhibitors of HER2.In some embodiments, the combination comprises at least one of the HER2 targeting STING agonist antibody-drug conjugates of the present disclosure combined with tucatinib, neratinib, or lapatinib.

[0125] In some embodiments, a combination comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one small molecule inhibitor of HER2, such as, for example, tucatinib, neratinib, or lapatinib.

[0126] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of a HER2-targeted therapeutic agent.

[0127] Immunotherapy Any suitable immunotherapeutic agent is contemplated herein for use in the combinations and methods of the present disclosure.In some aspects, the immunotherapeutic agent is an immune checkpoint inhibitor, including but not limited to immune checkpoint molecule binding proteins, small molecule inhibitors, antibodies, antibody derivatives (including Fab fragments and scFv), antibody-drug conjugates, antisense oligonucleotides, siRNA, aptamers, peptides, and peptidomimetics.Inhibitory nucleic acids that reduce the expression and / or activity of immune checkpoint molecules can also be used in the combinations and methods disclosed herein.

[0128] In some embodiments, immune checkpoint inhibitors reduce the expression or activity of one or more immune checkpoint proteins.In another embodiment, immune checkpoint inhibitors reduce the interaction between one or more immune checkpoint proteins and their ligands.See, for example, US20160101128.

[0129] In some aspects, immune checkpoint inhibitors suitable for the combinations and methods of the present disclosure are monoclonal antibodies, humanized antibodies, fully human antibodies, fusion proteins, or combinations thereof.

[0130] In some embodiments, the composition comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with at least one immune checkpoint inhibitor. In some embodiments, the composition comprises at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof.

[0131] In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

[0132] In some embodiments, the PD-1 or PD-L1 inhibitor is pembrolizumab (MK-3475), nivolumab (BMS-936558), pidilizumab (CT-011), AMP-224, MDX-1 105, durvalumab (MEDI4736), MPDL3280A, BMS-936559, IPH2101, TSR-042, TSR-022, cemiplimab, ipilimumab, lirilumab, atezolizumab, avelumab, dostallimab, tremelimumab, or a combination thereof.

[0133] In some aspects, the immune checkpoint inhibitor is avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab.

[0134] In some aspects, the immune checkpoint inhibitor is pembrolizumab, dostallimab, nivolumab, or atezolizumab. In some aspects, the immune checkpoint inhibitor is pembrolizumab. In some aspects, the immune checkpoint inhibitor is dostallimab.

[0135] In some embodiments, the combination comprises at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure and at least one immune checkpoint inhibitor. In some embodiments, the combination comprises at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure and at least one PD-1 inhibitor or at least one PD-L1 inhibitor.

[0136] In some embodiments, the combination comprises at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure and avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab. In some embodiments, the combination comprises at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure and dostallimab. In some embodiments, the combination comprises at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure and pembrolizumab.

[0137] In some embodiments, a combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one immune checkpoint inhibitor.

[0138] In some embodiments, a combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one PD-1 inhibitor or at least one PD-L1 inhibitor.

[0139] In some embodiments, the combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab.In some embodiments, the combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dostallimab.In some embodiments, the combination comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with pembrolizumab.

[0140] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of immunotherapeutic agents (e.g., immune checkpoint inhibitors).

[0141] How to use In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0142] In some aspects, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0143] In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0144] In some aspects, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0145] In some aspects, the present disclosure provides a method of activating or enhancing STING activity in a subject, the method comprising administering to the subject a combination therapy disclosed herein, i.e., at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0146] In some aspects, the present disclosure relates to a method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a combination therapy disclosed herein.

[0147] In some aspects, the present disclosure relates to a method of treating cancer in a subject in need thereof comprising administering to the subject a combination therapy disclosed herein.

[0148] In some aspects, the present disclosure provides a combination therapy as disclosed herein for use to treat or prevent a disease or disorder in a subject in need thereof.

[0149] In some aspects, the present disclosure provides a combination therapy as disclosed herein for use to treat a disease or disorder in a subject in need thereof.

[0150] In some aspects, the present disclosure provides a combination therapy as disclosed herein for treating a STING-mediated disease or disorder in a subject.

[0151] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein to treat or prevent a disease or disorder in a subject in need thereof.

[0152] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein to treat a disease or disorder in a subject in need thereof.

[0153] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein to treat cancer in a subject in need thereof.

[0154] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein for treating a STING-mediated disease or disorder in a subject in need thereof.

[0155] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein in the manufacture of a medicament for treating a disease or disorder in a subject in need thereof.

[0156] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein in the manufacture of a medicament for treating or preventing a disease or disorder in a subject in need thereof.

[0157] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein in the manufacture of a medicament for treating a STING-mediated disease or disorder in a subject.

[0158] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein in the manufacture of a medicament for treating cancer in a subject in need thereof.

[0159] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein for the treatment or prevention of a disease or disorder in a subject in need thereof.

[0160] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein for the treatment of a disease or disorder in a subject in need thereof.

[0161] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein to treat a STING-mediated disease or disorder in a subject.

[0162] In some aspects, the present disclosure provides for the use of a combination therapy disclosed herein for the treatment of cancer in a subject in need thereof.

[0163] In some embodiments, the combination comprises at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor.

[0164] In some aspects, a combination therapy disclosed herein is administered to the subject.

[0165] In some aspects, the present disclosure provides a method of treating or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2 targeted therapeutic agent or at least one immune checkpoint inhibitor; wherein the HER2-targeted STING agonist antibody-drug conjugate releases the one or more therapeutic agents upon biodegradation.

[0166] In some aspects, the present disclosure provides a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of at least one conjugate of the present disclosure; wherein the HER2-targeted STING agonist antibody-drug conjugate releases one or more therapeutic agents upon biodegradation.

[0167] In some aspects, the disease or disorder is cancer.

[0168] In some embodiments, the cancer is selected from the group consisting of anal cancer, astrocytoma, leukemia, lymphoma, head and neck cancer, liver cancer, testicular cancer, cervical cancer, sarcoma, hemangioma, esophageal cancer, eye cancer, laryngeal cancer, mouth cancer, mesothelioma, skin cancer, myeloma, oral cancer, rectal cancer, colorectal cancer (CRC), throat cancer, bladder cancer, breast cancer, urothelial cancer, uterine cancer, ovarian cancer, prostate cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, pancreatic cancer, renal cancer, gastric cancer, and gastroesophageal junction cancer.

[0169] In some embodiments, the cancer is selected from the group consisting of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0170] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0171] In some embodiments, the combination therapy disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of breast cancer. In some embodiments, the breast cancer is metastatic breast cancer. In some embodiments, the breast cancer is HER2 positive (HER2+) breast cancer. In some embodiments, the breast cancer is HER2 negative (HER2-) breast cancer. In some embodiments, the HER2+ breast cancer is metastatic HER2+ breast cancer. In some embodiments, the HER2- breast cancer is metastatic HER2- breast cancer.

[0172] In some embodiments, the subject with breast cancer has had at least one, at least two, at least three, or at least four lines of breast cancer therapy in the past. In some aspects, the subject has had three or more lines of breast cancer therapy in the past. In some aspects, the subject with HER2+ metastatic breast cancer has had three or more lines of breast cancer therapy.

[0173] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of gastric cancer.

[0174] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of gastroesophageal junction cancer.

[0175] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of colorectal cancer.

[0176] In some aspects, the combination therapies disclosed herein are useful for treating, preventing, delaying the progression of, or otherwise ameliorating the symptoms of non-small cell lung cancer (NSCLC).

[0177] In some embodiments, the subject has a recurrent or metastatic solid tumor with HER2+ expression.

[0178] In some aspects, the disease or disorder is a precancerous syndrome.

[0179] In some aspects, combination therapies comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2 targeted therapeutic agent or at least one immune checkpoint inhibitor are useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0180] In some aspects, combination therapies comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2 targeted therapeutic agent are useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0181] In some aspects, combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one immune checkpoint inhibitor is useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0182] In some aspects, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate is administered in combination with at least one HER2 antibody, at least one HER2 dimerization inhibitor antibody, or a combination of at least one HER2 antibody and at least one HER2 dimerization inhibitor antibody to treat, prevent, delay the progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0183] In some embodiments, combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate is administered in combination with trastuzumab or pertuzumab, or a combination thereof, or margetuximab, to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0184] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one biosimilar trastuzumab or at least one biosimilar pertuzumab, or a combination thereof, or at least one biosimilar margetuximab to treat, prevent, delay progression, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0185] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one HER2-targeted antibody-drug conjugate, such as ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®), to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0186] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with trastuzumab or a biosimilar of trastuzumab to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with ado-trastuzumab emtansine (T-DM1) (Kadcyla®) or a biosimilar of ado-trastuzumab emtansine (T-DM1) (Kadcyla®) to treat, prevent, delay progression, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) or a biosimilar of fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0187] In some embodiments, combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with trastuzumab or a biosimilar of trastuzumab to treat, prevent, delay progression, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer.In some embodiments, combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with trastuzumab or a biosimilar of trastuzumab to treat, prevent, delay progression, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer in subjects who have previously received at least one line, at least two lines, at least three lines, or at least four lines of breast cancer therapy.

[0188] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with a biosimilar of trastuzumab emtansine (T-DM1) (Kadcyla®) or ado-trastuzumab emtansine (T-DM1) (Kadcyla®) to treat, prevent, delay progression, or otherwise ameliorate symptoms of HER2-positive (HER2+) metastatic breast cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with a biosimilar of trastuzumab emtansine (T-DM1) (Kadcyla®) or ado-trastuzumab emtansine (T-DM1) (Kadcyla®) to treat, prevent, delay progression of, or otherwise ameliorate symptoms of HER2-positive (HER2+) metastatic breast cancer in a subject who has previously received at least one, at least two, at least three, or at least four lines of breast cancer therapy.

[0189] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) or a biosimilar of fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) to treat, prevent, delay the progression of, or otherwise ameliorate the symptoms of HER2-positive (HER2+) metastatic breast cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) or a biosimilar of fam-trastuzumab deruxtecan (trastuzumab deruxtecan) (Enhertu®) to treat, prevent, delay the progression of, or otherwise ameliorate the symptoms of HER2-positive (HER2+) metastatic breast cancer in a subject who has previously received at least one, at least two, at least three, or at least four lines of breast cancer therapy.

[0190] In some embodiments, combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one small molecule inhibitor of HER2, such as, for example, tucatinib, neratinib, or lapatinib, to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0191] In some aspects, a composition therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure in combination with at least one immune checkpoint inhibitor is useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0192] In some embodiments, a composition therapy comprises at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure in combination with a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof, and is useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0193] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one immune checkpoint inhibitor to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0194] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one PD-1 inhibitor or at least one PD-L1 inhibitor to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0195] In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with avelumab, durvalumab, dostallimab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab to treat, prevent, delay progression, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dostallimab to treat, prevent, delay progression, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer. In some embodiments, a combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate of the present disclosure is administered in combination with pembrolizumab to treat, prevent, delay progression of, or otherwise ameliorate symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0196] In some embodiments, a combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with dostallimab to treat, prevent, delay progression of, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer.In some embodiments, a combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with dostallimab to treat, prevent, delay progression of, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer in subjects who have previously received at least one line, at least two lines, at least three lines, or at least four lines of breast cancer therapy.

[0197] In some embodiments, a combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with pembrolizumab to treat, prevent, delay progression of, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer. In some embodiments, a combination therapy comprising at least one of the HER2-targeting STING agonist antibody-drug conjugates of the present disclosure is administered in combination with pembrolizumab to treat, prevent, delay progression of, or otherwise improve symptoms of HER2-positive (HER2+) metastatic breast cancer in subjects who have previously received at least one line, at least two lines, at least three lines, or at least four lines of breast cancer therapy.

[0198] In some embodiments, the method includes identifying or otherwise narrowing, e.g., stratifying, a patient population suitable for therapeutic administration of a combination therapy disclosed herein, by determining a subject's HER2 score by IHC testing or FISH (fluorescent in situ hybridization negative) measurements prior to treatment with the combination therapy disclosed herein. IHC testing measures the amount of HER2 receptor protein on the cell surface of a cancer tissue sample, e.g., a breast cancer tissue sample or a gastric cancer sample, and assigns a HER2 score of 0, 1+, 2+, or 3+ to the level of cell surface HER2 receptor detected. If a subject's HER2 score is in the range of 0-1+, the cancer is considered "HER2 negative." If the score is 2+, the cancer is referred to as "borderline," and a score of 3+ heralds the cancer as "HER2 positive."

[0199] In some embodiments, the subject is identified as having a score of 2+ or 3+ for HER2 expression detected by immunohistochemistry (IHC) analysis performed on the test cell population, and the subject is also ER positive. In some embodiments, the subject is identified as having a score of 2+ or 3+ for HER2 expression detected by immunohistochemistry (IHC) analysis performed on the test cell population, and the subject is also ER negative. In some embodiments, the subject is identified as having a score of 3+ for HER2 expression, or evidence of gene amplification by FISH. In some embodiments, the subject is identified as having a score of 2+ for HER2 expression, or evidence of gene amplification by FISH. In some embodiments, the subject is identified as having a score of 1+ for HER2 expression, or evidence of gene amplification by FISH. In some embodiments, the subject is identified as having high HER2 expression. In some embodiments, the subject is identified as having low HER2 expression. In some embodiments, the test cell population is derived from fresh, non-frozen tissue from a biopsy sample. In some aspects, the test cell population is derived from frozen tissue from a biopsy sample.

[0200] In some embodiments, the combination therapy comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immune checkpoint inhibitor disclosed herein is useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, colorectal cancer, or non-small cell lung cancer (NSCLC) in patients with HER2 IHC 2+ or HER2 IHC 3+. In some embodiments, the breast cancer patient is also ER positive. In some embodiments, the breast cancer patient is also ER negative.

[0201] In some aspects, combination therapies comprising at least one HER2-targeted STING agonist antibody-drug conjugate disclosed herein and at least one HER2 targeted therapeutic agent or at least one immune checkpoint inhibitor are useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, colorectal cancer, or non-small cell lung cancer (NSCLC) in patients with HER2 IHC 1+ or HER2 IHC 2+.

[0202] In some aspects, combinations comprising a HER2-targeted STING agonist antibody-drug conjugate disclosed herein and at least one HER2 targeted therapeutic agent or at least one immune checkpoint inhibitor are useful for treating, preventing, delaying the progression of, or otherwise ameliorating symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, colorectal cancer, or non-small cell lung cancer (NSCLC) in patients with HER2 IHC 1+ or HER2 IHC 2+.

[0203] In some embodiments, the combination comprising HER2 targeting STING agonist antibody-drug conjugate and at least one HER2 targeting therapeutic agent or at least one immune checkpoint inhibitor used in any of the methods and use embodiments provided herein can be administered at any stage of disease.For example, combination therapy can be administered to patients suffering from any stage of cancer, from early stage to metastasis.

[0204] The combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate and at least one HER2-targeting therapeutic agent or at least one immune checkpoint inhibitor used in any of these methods and use embodiments can be administered either without another therapeutic agent or in further combination with one or more chemotherapeutic agents or other agents.In some embodiments, the additional agent is any of the toxins described herein.In some embodiments, the additional agent is (1) an EGFR inhibitor (e.g., a tyrosine kinase inhibitor or a targeted anti-EGFR antibody), (2) a BRAF inhibitor, (3) an ALK inhibitor, (4) a hormone receptor inhibitor, (5) an mTOR inhibitor, (6) a VEGF inhibitor, or (7) a cancer vaccine. In some embodiments, the additional agent is, for example, ado-trastuzumab emtansine (Kadcyla), lapatinib, anastrozole, letrozole, exemestane, everolimus, fulvestrant, tamoxifen, toremifene, megestrol acetate, fluoxymesterone, ethinyl estradiol, paclitaxel, capecitabine, gemcitabine, eribulin, vinorelbine, cyclophosphamide, carboplatin, docetaxel, albumin-bound paclitaxel, cisplatin, epirubicin. In some embodiments, the additional agent is Kadcyla (ado-trastuzumab emtansine).

[0205] Combination Therapies and Formulations The combination therapies of the present disclosure can be administered together in a single pharmaceutical composition or separately, and when administered separately, can be administered simultaneously or sequentially in any order. The amounts of the components of the combinations of the present disclosure and the relative timings of administration are selected in order to achieve the desired combined therapeutic effect.

[0206] The compositions of the present disclosure may be administered once or according to a dosing regimen in which several doses are administered at various time intervals over a given period of time. For example, doses may be administered daily, weekly, biweekly, monthly, bimonthly, every six months, or annually. Doses may be administered until a desired therapeutic effect is achieved, or indefinitely to maintain a desired therapeutic effect. A suitable dosing regimen for a composition of the present disclosure depends on the pharmacokinetic properties of the composition, such as absorption, distribution, and half-life, which can be determined by those skilled in the art. In addition, a suitable dosing regimen for a composition of the present disclosure depends on the disease or disorder being treated, the severity of the disease or disorder being treated, the age and physical condition of the patient being treated, the medical history of the patient being treated, the nature of concurrent therapy, the desired therapeutic effect, and similar factors within the knowledge and expertise of those skilled in the art. It will be further understood by such skilled artisans that adjustments of a suitable dosing regimen may be necessary in view of an individual patient's response to the dosing regimen, or over time as the needs of an individual patient change.

[0207] It will be appreciated that administration of the conjugates and HER2 targeted therapeutic agents or immune checkpoint inhibitors in the combinations disclosed herein will be administered with suitable carriers, excipients, and other agents that are incorporated into the formulation to provide improved transport, delivery, tolerability, etc. Many suitable formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th ed., Mack Publishing Company, Easton, PA (1975)), particularly Chapter 87 by Blaug, Seymour.

[0208] For example, the combination therapy may include one or more conjugates disclosed herein co-formulated and / or co-administered with one or more HER2 targeted therapeutic agents or one or more immune checkpoint inhibitors disclosed herein.

[0209] In some embodiments, pharmaceutical composition is in bulk or unit dosage form. Unit dosage form can be in any of a variety of forms, including, for example, capsule, IV bag, tablet, single pump in aerosol inhaler, or vial. The amount of active ingredient (e.g., conjugate disclosed herein) in unit dosage of composition is an effective amount and varies according to the specific treatment involved. Those skilled in the art will know that sometimes it is necessary to vary dosage routinely depending on the age and condition of the patient. Dosage also depends on the route of administration.

[0210] A pharmaceutical composition is formulated to be compatible with its intended route of administration. Exemplary routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (i.e., topical), transmucosal, and rectal administration.

[0211] In some embodiments, pharmaceutical composition is in bulk or unit dosage form. Unit dosage form can be in any of a variety of forms, including, for example, capsule, IV bag, tablet, single pump in aerosol inhaler, or vial. The amount of active ingredient (e.g., conjugate disclosed herein) in unit dosage of composition is effective amount and varies according to the specific treatment involved. Those skilled in the art will know that sometimes it is necessary to vary dosage routinely depending on the age and condition of the patient. Dosage also depends on the route of administration.

[0212] For example, the combination therapy may include one or more conjugates disclosed herein co-formulated and / or co-administered with one or more HER2-targeted therapeutic agents or one or more immune checkpoint inhibitors disclosed herein.

[0213] In some embodiments, the combination comprising HER2-targeted STING agonist antibody-drug conjugate, HER2-targeted therapeutic agent or immune checkpoint inhibitor, and additional agent is formulated into a single therapeutic composition, and the components are administered simultaneously. Alternatively, HER2-targeted STING agonist antibody-drug conjugate, HER2-targeted therapeutic agent or immune checkpoint inhibitor, and additional agent, if present, are separate from each other, for example, each can be formulated into separate therapeutic composition and administered simultaneously, or can be administered at different times during treatment regimen. For example, HER2-targeted STING agonist antibody-drug is administered before administration of HER2-targeted therapeutic agent or immune checkpoint inhibitor; HER2-targeted STING agonist antibody-drug is administered after administration of HER2-targeted therapeutic agent or immune checkpoint inhibitor. As described herein, the combination of HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor is administered in a single dose or multiple doses.

[0214] Dosage and Administration The combination therapy provided herein, comprising the combination of HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor, administered in a single dose or multiple doses, is administered in an amount sufficient to exert therapeutically useful effects.In some embodiments, the active agent is administered in an amount that does not cause undesirable side effects to the treated patient, or in an amount that minimizes or reduces the side effects observed compared to the dosage and amount required for a single treatment with one of the above-mentioned agents.For example, the combination therapy comprising the combination of HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor is administered in a single dose or multiple doses.Therefore, the amount of HER2-targeted therapeutic agent or immune checkpoint inhibitor that can be administered in the combination therapy provided herein can be reduced while achieving substantially the same or improved therapeutic efficacy compared to the amount of HER2-targeted therapeutic agent or immune checkpoint inhibitor administered alone or using known methods. The reduced dosage that can be administered reduces, minimizes, or avoids side effects associated with administration of HER2 targeted therapeutic agents or immune checkpoint protein antibodies, such as the immune-related adverse events described elsewhere or herein.

[0215] It is within the level of a person skilled in the art to determine the exact amount of the HER2-targeting STING agonist antibody-drug conjugate and the active agent, including the HER2-targeting therapeutic agent or immune checkpoint inhibitor, to be administered to a subject.The dosage of such agents in combination therapy can be selected based on the standard dosage regimen of the agent under a given route of administration.

[0216] It is understood that the exact dosage and duration of treatment is a function of the tissue or tumor being treated, and can be determined empirically using known testing protocols, or by extrapolation from in vivo or in vitro testing data, and / or can be determined from known dosing regimens of a particular agent. It is noted that concentration and dosage values ​​can also vary according to the age of the individual being treated, the weight of the individual, the route of administration, and / or the extent or severity of disease, as well as other factors within the level considered by a skilled physician. In general, the dosage regimen is selected to limit toxicity. It should be noted that the treating physician knows how and when to terminate, interrupt, or adjust therapy to reduce dosage due to toxicity, or bone marrow, liver, or kidney, or other tissue dysfunction. Conversely, the treating physician also knows how and when to adjust treatment to a higher level (while avoiding toxic side effects) if clinical response is not adequate. It is further understood that for any particular subject, a specified dosage regimen will be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the formulations, and that the concentration ranges set forth herein are exemplary only and are not intended to be limiting in scope.

[0217] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugate combination is administered in a therapeutically effective amount to reduce tumor volume.

[0218] The amount of HER2 targeting STING agonist antibody-drug conjugate administered for the treatment of disease or condition, such as cancer or solid tumor, can be determined by standard clinical techniques.In addition, in vitro assays and animal models can be adopted to help identify optimal dosage range.The exact dosage that can be empirically determined can depend on the route of administration, the type of disease being treated, and the severity of the disease.

[0219] The HER2 targeting therapeutic agent or immune checkpoint inhibitor can be provided in a therapeutically effective amount for a specific dosage regimen.The therapeutically effective concentration can be empirically determined by testing the compound in known in vitro and in vivo systems, such as the assays provided herein.The concentration of the selected HER2 targeting therapeutic agent relative to the immune checkpoint inhibitor in the composition depends on the absorption, inactivation and excretion rate of the complex, the physicochemical characteristics of the complex, the dosage schedule and the amount administered, and other factors known to those skilled in the art.

[0220] The amount of the selected HER2 targeting therapeutic agent or immune checkpoint inhibitor administered for the treatment of cancer can be determined by standard clinical techniques or other methods described herein.In addition, in vitro assays and animal models can be adopted to help identify optimal dosage ranges.Accurate dosages that can be empirically determined can therefore depend on the route of administration, the type of cancer being treated, and disease progression.If necessary, specific dosages and durations and treatment protocols can be empirically determined or extrapolated.Dosage levels can be determined based on a variety of factors, such as individual weight, general health, age, the activity of the specified compound being employed, sex, diet, administration time, excretion rate, drug combination, disease severity and course, and patient's disposition to disease, as well as the judgment of the treating physician.

[0221] The combinations of the present disclosure may be administered by any suitable route of administration, including both systemic and local administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to a route of administration other than enteral, transdermal, or by inhalation, such as by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration to the patient's lungs, whether inhaled through the mouth or through the nasal passages. Local administration includes application to the skin.

[0222] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugate and the HER2 targeted therapy or immune checkpoint inhibitor are administered as an infusion every 1, 2, 3, 4, 5, 6, 7, or 8 weeks.

[0223] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugate and the HER2 targeted therapeutic agent or immune checkpoint inhibitor are administered as an infusion every three or four weeks.

[0224] In some embodiments, HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor are administered simultaneously by injection.In some embodiments, HER2-targeted STING agonist antibody-drug is administered by injection before administration of HER2-targeted therapeutic agent or immune checkpoint inhibitor.In some embodiments, HER2-targeted STING agonist antibody-drug is administered by injection after administration of HER2-targeted therapeutic agent or immune checkpoint inhibitor.As described herein, the combination of HER2-targeted STING agonist antibody-drug and HER2-targeted therapeutic agent or immune checkpoint inhibitor can be administered in a single dose or multiple doses.

[0225] The frequency and timing of administration, and the amount of dosage can be periodically controlled over a cycle of administration to maintain the continuous and / or long-term effect of active agent for a desired period of time.The combination provided can be administered hourly, daily, weekly, monthly, yearly, or once.The duration of a cycle of administration can be empirically determined and depends on the disease being treated, the severity of the disease, the specific patient, and other considerations within the skill level of the treating physician.The duration of treatment with the combination therapy provided herein can be one week, two weeks, one month, several months, one year, several years, or longer.

[0226] In some aspects, the exemplary doses of immune checkpoint inhibitors, such as anti-immune checkpoint protein antibodies, administered intravenously can be used as a starting point to determine appropriate dosages. Dosage levels can be determined based on a variety of factors, such as the individual's weight, general health, age, activity of the specified compound employed, sex, diet, time of administration, excretion rate, drug combination, severity and course of the disease, and patient disposition due to the disease, as well as the judgment of the treating physician.

[0227] It is understood that the amount administered will be a function of the type of cancer being treated, the route of administration, and the acceptability of possible side effects. If necessary, dosages can be empirically determined.

[0228] For intravenous administration, one or more or all of the agents used in the combination therapy may be administered by push or bolus, by infusion, or via a combination thereof. The infusion time may be from about 1 minute to about 3 hours, such as from about 1 minute to about 2 hours, from about 1 minute to about 60 minutes, from about 1 minute to about 90 minutes, or from about 1 minute to about 120 minutes. The agents may be administered by parallel infusion or by subsequent infusion. For example, the agents to be administered are administered separately and provided in separate bags for separate infusions. In some embodiments, the HER2-targeted antibody-drug conjugate composition and the HER-2 targeted therapeutic agent or immune checkpoint inhibitor composition are formulated and administered separately.

[0229] The HER2-targeting STING agonist antibody-drug conjugate can be administered prior to, simultaneously or nearly simultaneously with, sequentially, or intermittently with the HER2-targeting therapeutic agent or immune checkpoint inhibitor. For example, the HER2-targeting antibody-drug conjugate and the HER2-targeting therapeutic agent or immune checkpoint inhibitor, such as an anti-immune checkpoint protein antibody (e.g., anti-CTLA4 or anti-PD-1 antibody), can be co-administered or administered separately.

[0230] In some embodiments, the HER2 targeting STING agonist antibody-drug conjugate is administered before the HER2 targeting therapeutic agent or immune checkpoint inhibitor.For example, the HER2 targeting STING agonist antibody-drug conjugate is administered up to about 48 hours before the HER2 targeting therapeutic agent or immune checkpoint inhibitor is administered.For example, the HER2 targeting STING agonist antibody-drug conjugate is administered about 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 40 hours, or up to about 48 hours before the HER2 targeting therapeutic agent or immune checkpoint inhibitor is administered.

[0231] In some embodiments, the HER2 targeting STING agonist antibody-drug conjugate is administered after the HER2 targeting therapeutic agent or immune checkpoint inhibitor.For example, the HER2 targeting STING agonist antibody-drug conjugate is administered at most about 48 hours after the administration of the HER2 targeting therapeutic agent or immune checkpoint inhibitor.For example, the HER2 targeting STING agonist antibody-drug conjugate is administered at most about 5 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 12 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 30 hours, 36 hours, 40 hours, or at most about 48 hours after the administration of the HER2 targeting therapeutic agent or immune checkpoint inhibitor.

[0232] The frequency and timing of administration, as well as the amount of dosage, can be periodically managed over an administration cycle to maintain the continuous and / or long-term effect of the active agent for a desired period, and need not be the same for the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapeutic agent or immune checkpoint inhibitor. The compositions of each active agent provided or their combinations can be administered hourly, daily, weekly, monthly, yearly, or once. The duration of an administration cycle can be empirically determined and depends on the disease being treated, the severity of the disease, the patient's disposition, and other considerations within the skill level of the treating physician. The duration of treatment with the combination therapy provided herein can be one week, two weeks, one month, several months, one year, several years, or longer.

[0233] For example, the administration frequency of the HER2-targeting STING agonist antibody-drug conjugate is once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once every four weeks. The dosage can be divided into multiple administration cycles during the course of treatment. For example, the HER2-targeting STING agonist antibody-drug conjugate can be administered at a frequency that lasts for about one month, two months, three months, four months, five months, six months, one year, or more. The administration frequency can be the same or different throughout the cycle period. For example, an exemplary dosage frequency is twice a week for at least the first week of the administration cycle. After the first week, the frequency can continue twice a week, can be increased to more than twice a week, or can be reduced to once a week or less. It is within the level of one of ordinary skill in the art to determine the specific dosage frequency and administration cycle based on the specific dosage being administered, the disease or condition being treated, the severity of the disease or condition, the age of the subject, and other similar factors.

[0234] The HER2-targeted therapy or immune checkpoint inhibitor may be administered at the same frequency or at different frequencies. For example, administration of the HER2-targeted STING agonist antibody-drug conjugate precedes each administration of the HER2-targeted therapy or immune checkpoint inhibitor within 48 hours. For example, each dose of the HER2-targeted STING agonist antibody-drug conjugate is followed by a dose of the HER2-targeted therapy or immune checkpoint inhibitor 24-48 hours later. In certain embodiments, the HER2-targeted therapy or immune checkpoint inhibitor is administered less frequently than the HER2-targeted STING agonist antibody-drug conjugate, but each dose of the HER2-targeted therapy or immune checkpoint inhibitor is preceded by a dose of the HER2-targeted antibody-drug conjugate. For example, the HER2 targeting therapy or immune checkpoint inhibitor is administered twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every six weeks, once every two months, once every three months, once every four months, once every five months, or once every six months, and is administered in a manner that precedes the administration of the HER2 targeting STING agonist antibody-drug conjugate.In another example, a dose of the HER2 targeting therapy or immune checkpoint inhibitor precedes each dose of the HER2 targeting STING agonist antibody-drug conjugate.In certain embodiments, the HER2 targeting therapy or immune checkpoint inhibitor is administered more frequently than the HER2 targeting STING agonist antibody-drug conjugate. For example, the HER2 targeted therapy or immune checkpoint inhibitor is administered twice weekly, once weekly, once every two weeks, once every three weeks, once every four weeks, once every six weeks, once every two months, once every three months, once every four months, once every five months, or once every six months, and in a manner such that a portion, but not all, of the dosage of the HER2 targeted therapy or the dosage of the checkpoint inhibitor is followed by administration of the HER2 targeted STING agonist antibody-drug conjugate.

[0235] If disease symptoms persist in the absence of cessation of treatment, treatment may be continued for an additional period of time. Over the course of treatment, evidence of disease and / or treatment-related toxicity or side effects may be monitored.

[0236] The administration cycle of HER2 targeting STING agonist antibody-drug conjugate and / or HER2 targeting therapeutic agent and / or immune checkpoint inhibitor can be individually adjusted to add a period of treatment cessation to provide a rest period from exposure to the agent.The period of treatment cessation can be a predetermined time or can be empirically determined depending on how the patient responds or on observed side effects.For example, treatment can be halted for one week, two weeks, one month, or several months.Generally, the period of treatment cessation is incorporated into the cycle of the patient's dosing regimen.

[0237] Exemplary dosing regimen is a 21-day or 28-day treatment or administration cycle.The agent such as the HER2-targeting STING agonist antibody-drug conjugate disclosed herein can be administered on the first day, followed by the HER2-targeting therapeutic agent or immune checkpoint inhibitor of the present disclosure such as an immune checkpoint protein antibody on the second day, followed by no dosing for the next 19 or 26 days.It is understood that the above description is merely illustrative, and that the above variations can also be adopted.Furthermore, in the combination therapy provided herein, the same dosing cycle can be applied to all the administered agents, or each administered agent can adopt its own dosing regimen.

[0238] It is within the skill level of a practitioner to determine the correct administration cycle and dosing schedule.As mentioned above, the administration cycle can be for any desired period.Thus, the 21-day or 28-day administration cycle can be repeated for any period.Depending on the patient's specific individual considerations and the disease being treated, it is within the skill level of the treating physician to adopt the administration cycle and dosing regimen that meets the patient's needs.

[0239] Diagnostic and prophylactic products Conjugate and HER2 targeting therapeutic agent or immune checkpoint inhibitor disclosed herein are used in diagnostic and preventive preparations.In one embodiment, HER2 targeting STING agonist antibody-drug conjugate disclosed herein and HER2 targeting therapeutic agent or immune checkpoint inhibitor are administered to patients who are at risk of developing one or more of the above-mentioned diseases, such as but not limited to cancer.The predisposition of patients or organs to one or more of the above-mentioned indications can be determined using genotypic markers, serological markers, or biochemical markers.

[0240] In some embodiments, the HER2-targeted STING agonist antibody-drug conjugates and immune checkpoint inhibitors disclosed herein are administered to a human individual diagnosed with a clinical indication associated with one or more of the aforementioned diseases, such as, but not limited to, cancer. Once diagnosed, the HER2-targeted STING agonist antibody-drug conjugates disclosed herein and HER2-targeted therapeutic agents or immune checkpoint inhibitors are administered to alleviate or reverse the effects of the clinical indication associated with one or more of the aforementioned diseases.

[0241] In some embodiments, a method for identifying a breast cancer patient suitable for treatment with a combination of a conjugate disclosed herein and a HER2 targeted therapeutic agent or an immune checkpoint inhibitor comprises measuring the status of certain features in a tumor sample obtained from the patient, and identifying the patient for treatment based on the status of certain features in the tumor sample.

[0242] The antibody disclosed herein is also useful for detecting HER2 in patient samples, and is therefore useful as a diagnostic method.For example, the HER2 antibody disclosed herein is used in in vitro assays, such as ELISA, to detect HER2 levels in patient samples.

[0243] In some embodiments, the HER2 antibody disclosed herein is immobilized on a solid support (e.g., the well of a microtiter plate).The immobilized antibody serves as the capture antibody of any HER2 that may be present in the test sample.Before the immobilized antibody is contacted with the patient sample, the solid support is rinsed and treated with a blocking agent such as milk protein or albumin to prevent non-specific adsorption of analyte.

[0244] The well is then treated with a test sample suspected of containing the antigen or with a solution containing a standard amount of the antigen. Such a sample is, for example, a serum sample from a subject suspected of having a circulating antigen level considered diagnostic for a disease state. After rinsing off the test sample or standard, the solid support is treated with a detectably labeled second antibody. The labeled second antibody serves as a detection antibody. The level of detectable label is measured, and the concentration of HER2 antigen in the test sample is determined by comparison with a standard curve made from the standard sample.

[0245] It can be seen that, based on the results obtained by using the HER2 antibody disclosed herein in in vitro diagnostic assay, it is possible to determine the stage of disease in a subject based on the expression level of HER2 antigen.For a given disease, blood samples are taken from subjects diagnosed as being at different stages of disease progression and / or at different times during the therapeutic treatment of disease.Using the sample population that provides statistically significant results for each stage of progression or therapy, the range of concentrations of antigen that can be considered characteristic for each stage is defined.

[0246] All publications and patent documents cited in this specification are incorporated herein by reference as if each such publication or document was specifically and individually indicated to be incorporated herein by reference. Citation of publications and patent documents is not intended as an admission that any is relevant prior art, and does not constitute any admission as to its contents or date. Although the present invention has now been described by written description, those skilled in the art will recognize that the present invention can be practiced in various embodiments, and that the foregoing description and the following examples are for illustrative purposes and do not limit the scope of the following claims. EXAMPLES

[0247] The following examples are illustrative of the present disclosure.These examples are not intended to limit the scope of the present disclosure, but rather to provide guidance to those skilled in the art for preparing and using the compounds, compositions and methods of the present disclosure.Although specific embodiments of the present disclosure have been described, those skilled in the art will recognize that various changes and modifications can be made without departing from the spirit and scope of the present disclosure.

[0248] It will be appreciated that certain compounds of the present disclosure may be potent immune modulators and therefore care should be taken in their handling.

[0249] Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification.

[0250] Abbreviation The following abbreviations are used in the reaction schemes and synthetic examples which follow: This list is not meant to be a comprehensive list of abbreviations used in this application, as additional standard abbreviations readily understood by one of ordinary skill in the art of organic synthesis may also be used in the synthetic schemes and examples. TIFF2025509266000009.tif76128

[0251] General Information Unless otherwise stated, all reagents were purchased from relevant suppliers.

[0252] XMT-1519 (anti-Her2 antibody) is disclosed in US 9,555,112, issued January 31, 2017, and US 9,738,720, issued August 22, 2017, the entire contents of which are incorporated herein by reference.

[0253] The antibody-drug conjugate Enhertu is also known as trastuzumab deruxtecan or T-DXd.

[0254] The 7.16.4_msIgG2a mab is a murine IgG2a antibody that recognizes the p185 molecule (rat HER2(neu)) encoded by the rat neu oncogene. Conjugates 1 and 2 could not be used in the rat HER2 EMT6-RHER2-MSA breast adenocarcinoma tumor model because the humanized antibody XMT-1519 does not bind to rat HER2. Therefore, surrogate conjugates (7.16.4_msIgG2a conjugate and the non-binding control palivizumab_msIgG2a conjugate, conjugates 4 and 3, respectively) were used in the rat HER2 EMT6-RHER2-MSA breast adenocarcinoma tumor model because they specifically recognize rat HER2(neu).

[0255] HPLC purification was performed on a Phenomenex Gemini 5 μm C18 110 Å, 250×10 mm, semi-preparative column.

[0256] When applicable, the drug content of the conjugates was determined spectrophotometrically, otherwise RP-HPLC or LC / MS was performed for quantitative determination of drug content.

[0257] The protein content of the antibody-drug conjugates was determined spectrophotometrically or by ELISA.

[0258] The antibody-drug conjugate, drug-loaded scaffold, or antibody scaffold was purified (i.e., to remove residual unreacted drug, unconjugated antibody, enzyme, or starting material) by ultrafiltration, large-scale diafiltration, CHT chromatography, or HIC, as needed. If necessary, additional purification by SEC or HIC was performed to remove aggregated antibody-drug conjugate. In general, the purified antibody-drug conjugate contained <5% (w / w) (e.g., <2% (w / w)) aggregated antibody-drug conjugate as determined by SEC; <0.5% (w / w) (e.g., <0.1% (w / w)) free (unconjugated) drug as determined by RP-HPLC and / or LC-MS / MS; <1% (w / w) free drug conjugate as determined by SEC and / or RP-HPLC; and <10% (w / w) (e.g., <1% (w / w)) unconjugated antibody or antibody fragment as determined by HIC-HPLC and / or RP-HPLC. Reduced or partially reduced antibodies were prepared using procedures described in the literature. See, for example, Francisco et al., Blood 102 (4): 1458-1465 (2003). The concentration of total drug (conjugated and unconjugated) was determined by UV-Vis spectrophotometry or RP-HPLC.

[0259] The drug-to-antibody ratio (DAR) was determined by measuring the absorbance of the conjugates. DAR values ​​were calculated using the appropriate molar extinction coefficients of the antibody and STING agonist payloads.

[0260] Therapeutic agents, including HER2-targeted STING agonist ADCs, HER2-targeted therapeutic agents, and / or immunotherapeutic agents, can be administered or dosed at frequencies and intervals as disclosed herein. In some aspects, the therapeutic administration schedule can be as follows: Treatment schedule TIFF2025509266000010.tif46128

[0261] Antibody / payload dosage refers to mg of antibody per kilogram of body weight and mg of payload per kilogram of body weight. For example, a conjugate dosage of 0.30 / 0.011 mg / kg refers to administering 0.30 mg of antibody per kilogram of body weight and 0.011 mg of conjugate payload per kilogram of body weight.

[0262] Tumors were measured twice weekly using digital calipers and tumor volumes were calculated using the formula: tumor volume (mm 3 )=(width 2 Body weights were recorded daily for the first week and twice weekly thereafter. Animals were randomly assigned to receive a tumour with an individual tumour volume of ≥ 1000 mm 3 , ≧1500mm 3 Animals were kept on study until they reached the age of 18 or as indicated. Percentage change in body weight was calculated using the formula: Body Weight Change (%) = ((body weight on study day X - body weight 試験1日目 ) / body weight 試験1日目 )*100. Tumor volumes are reported as the mean ± standard error of the mean (SEM). Tumor growth inhibition (TGI%) was defined as the percentage difference in mean tumor volume (MTV) between treatment and control groups. To determine tumor growth inhibition (TGI), tumor size was measured throughout each efficacy study. Percent tumor regression was calculated using the formula: % Regression = (1-(Mean Tumor Volume 最終 ) / (mean tumor volume 1日目 A partial response (PR) was defined as a tumor volume that was 50% or less than the volume on day 1 on three consecutive measurements and ≥13.5 mm on at least one of these three measurements. 3 A complete response (CR) was defined as a tumor volume of 13.5 mm or greater on three consecutive measurements. 3 Tumor-free survivors (TFS) are classified as having a CR at the end of the study.

[0263] Example 1: Synthesis of XMT-1519 conjugate 1, DAR8.1 TIFF2025509266000011.tif79160 Conjugate 1 was prepared as described in co-pending application US17 / 221,341, filed April 4, 2021. Purified conjugate 1 had a STING agonist vs. XMT-1519 of 8.1.

[0264] Example 2: Synthesis of Palivizumab Conjugate 2, DAR6.8 TIFF2025509266000012.tif68142 Conjugate 2 was prepared as described in co-pending application US17 / 221,341, filed April 4, 2021. Purified conjugate 2 had a STING agonist to palivizumab ratio of 6.8.

[0265] Example 3: Synthesis of Palivizumab_msIgG2a conjugate 3, DAR7.7 TIFF2025509266000013.tif132144To a solution of Palivizumab_msIgG2a (90 mg, 0.617 μmol) in 50 mM HEPES, 1 mM EDTA, pH 7 buffer, TCEP-HCl (1.76 mg, 6.17 μmol) was added to a final antibody concentration of 5 mg / mL, and the mixture was shaken at 37 °C for 4 h. Scaffold A (prepared as described in co-pending application US17 / 221,341, filed April 4, 2021, 17.45 mg, 7.404 μmol in 1800 μL DMA) was added to the reduced antibody. The resulting mixture was shaken at 37 °C for 120 min. The reaction was quenched with cysteine ​​(15 equivalents, 1.121 mg, 9.255 μmol in 1.12 mL of 50 mM HEPES, 1 mM EDTA, pH 7) and rotated at room temperature for 1 h. The resulting conjugate 3 was purified by ultrafiltration or CHT chromatography (80 mg, 90% yield). The purified conjugate 3 had a STING agonist to palivizumab_msIgG2a ratio of 7.7.

[0266] Example 4: Synthesis of 7.16.4_msIgG2a conjugate 4, DAR6.9 TIFF2025509266000014.tif75160 Conjugate 4 was prepared and characterized as described in Example 3, except that 7.16.4_msIgG2a was used instead of Palivizumab_msIgG2a. Purified conjugate 4 had a STING agonist to 7.16.4_msIgG2a ratio of 6.9.

[0267] Example 5: Tumor growth response to administration of HER2 STING agonist antibody-drug conjugates in combination with trastuzumab in SKOV3 ovarian cancer Female CB.17 SCID mice were inoculated with SKOV3 human ovarian cancer cells (10 × 10 6 SKOV3 cells are a breast cancer cell line with high HER2 expression. 3 (Average=72~81mm 3 Animals were randomized into treatment groups when the dose was 100 mg / group. Vehicle, conjugate 1 (0.30 / 0.013 mg / kg), conjugate 1 (0.30 / 0.013 mg / kg) in combination with trastuzumab (3 mg / kg), conjugate 1 (0.30 / 0.013 mg / kg) in combination with trastuzumab (10 mg / kg), and conjugate 2 (0.30 / 0.011 mg / kg) in combination with trastuzumab (10 mg / kg) were dosed qd×1 on day 1 or qwk×3 on days 1, 8, and 15. Additionally, trastuzumab (10 mg / kg) was dosed qwk×3 starting on days 1, 8, and 15. For all doses, conjugates were dosed intravenously (IV); trastuzumab was dosed intraperitoneally (IP); all doses of conjugates are reported as antibody / payload; n=10 per group. FIG. 1 provides tumor volume results for SKOV3 tumor-bearing mice treated with vehicle, conjugate 1, trastuzumab, a combination of conjugate 2 and trastuzumab, and a combination of conjugate 1 and trastuzumab at various dose levels and dosing regimens. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd×1) resulted in one PR and three CRs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd×1) and trastuzumab (3 mg, qd×1) resulted in seven CRs and three TFSs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd×1) and trastuzumab (10 mg, qd×1) resulted in two CRs and one TFS. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qwk x 3) resulted in 4 CRs and 2 TFSs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qwk x 3) and trastuzumab (3 mg, qwk x 3) resulted in 1 PR, 9 CRs, and 9 TFSs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qwk x 3) and trastuzumab (10 mg, qwk x 3) resulted in 2 PRs, 7 CRs, and 7 TFSs. The results indicate that the addition of trastuzumab synergistically increases the efficacy of conjugate 1.

[0268] Example 6: Tumor growth response to administration of HER2 STING agonist antibody-drug conjugates in combination with trastuzumab or pertuzumab in JIMT-1 Female CB.17 SCID mice were inoculated with JIMT-1 tumor cells (10 × 10 6 JIMT-1 cells are a breast cancer cell line with moderate HER2 expression. 3 (Average 67.8~71.6mm 3Animals were randomized into treatment groups when the number of treatments per group was 100:1 (0.001 mg / kg / group) as follows: vehicle, conjugate 1 (3.0 / 0.13 mg / kg), conjugate 2 (3.0 / 0.11 mg / kg) in combination with trastuzumab (10 mg / kg), conjugate 2 (3.0 / 0.11 mg / kg) in combination with pertuzumab (10 mg / kg), conjugate 1 (3.0 / 0.13 mg / kg) in combination with trastuzumab (3 mg / kg), conjugate 1 (3.0 / 0.13 mg / kg) in combination with trastuzumab (10 ... Combinations of conjugate 1 (3.0 / 0.13 mg / kg) and pertuzumab (3 mg / kg), conjugate 2 (3.0 / 0.11 mg / kg), trastuzumab (3 mg / kg), and pertuzumab (3 mg / kg), and conjugate 1 (3.0 / 0.13 mg / kg), trastuzumab (3 mg / kg), and pertuzumab (3 mg / kg) were dosed qd×1 on day 1. Additionally, conjugate 1 (0.30 / 0.013 mg / kg), conjugate 2 (0.30 / 0.011 mg / kg) in combination with trastuzumab (3 mg / kg), conjugate 2 (0.30 / 0.011 mg / kg) in combination with pertuzumab (3 mg / kg), conjugate 1 (0.30 / 0.013 mg / kg) in combination with trastuzumab (3 mg / kg), and conjugate 1 (0.30 / 0.013 mg / kg) in combination with pertuzumab (3 mg / kg) were dosed qwk×3 on days 1, 8, and 15. For all doses, conjugates were dosed intravenously; trastuzumab and pertuzumab were dosed intraperitoneally; all doses of conjugates are reported as antibody / payload; n=10 for each group.

[0269] FIG. 2 provides the tumor volume results of JIMT-1 tumor-bearing mice treated with vehicle, conjugate 1, combination of conjugate 1 with trastuzumab, combination of conjugate 1 with pertuzumab, combination of conjugate 1 with trastuzumab and pertuzumab, combination of conjugate 2 with trastuzumab, combination of conjugate 2 with pertuzumab, and combination of conjugate 2 with trastuzumab and pertuzumab at various dose levels and dosing regimens.

[0270] All treatment groups showed significant tumor growth delay (TGD) compared to vehicle controls. However, only the group treated with conjugate 1 (3 mg / kg) alone or in combination with trastuzumab and / or pertuzumab, and the group treated with conjugate 1 (0.3 mg / kg) in combination with trastuzumab or pertuzumab achieved the maximum possible TGD (78%), although the groups produced different numbers of regression responses and tumor-free survivors (TFS). Treatment with conjugate 1 (3 mg / kg, qd x 1) and pertuzumab (3 or 10 mg / kg, qd x 1) resulted in 9 and 8 TFS, respectively, and 10 CRs each. Treatment with conjugate 1 (3 mg / kg, qd x 1) and trastuzumab (3 or 10 mg / kg, qd x 1) resulted in 3 TFS each, 10 CR, and 9 CR and 1 PR, respectively. All treatment groups except one, conjugate 2 with trastuzumab and pertuzumab, showed significant TGI compared to vehicle controls.

[0271] Example 7: Tumor growth response to administration of HER2 STING agonist antibody-drug conjugates in combination with trastuzumab or pertuzumab in SNU-5 Female CB.17 SCID mice were inoculated with SNU-5 tumor cells (10 × 10 6 SNU-5 is a gastric cancer cell line with low HER2 expression.3 (Average 79.7~81.3mm 3 Animals were randomized into treatment groups when the dose was 100 / group. Vehicle, conjugate 1 (0.20 / 0.007mg / kg), trastuzumab (2mg / kg), pertuzumab (2mg / kg), conjugate 2 (0.20 / 0.007mg / kg), a combination of conjugate 1 (0.20 / 0.007mg / kg) and trastuzumab (2mg / kg), a combination of conjugate 1 (0.20 / 0.007mg / kg) and pertuzumab (2mg / kg), a combination of trastuzumab (2mg / kg) and pertuzumab (2mg / kg), and a combination of conjugate 1 (0.20 / 0.007mg / kg), trastuzumab (2mg / kg), and pertuzumab (2mg / kg) were dosed qd×1 on day 1. Additionally, for conjugate 1 (0.10 / 0.004 mg / kg), conjugate 2 (0.10 / 0.004 mg / kg), the combination of conjugate 1 (0.10 / 0.004 mg / kg) with trastuzumab (2 mg / kg), and the combination of conjugate 1 (0.10 / 0.004 mg / kg) with pertuzumab (2 mg / kg), conjugates were dosed qwk×3 on days 1, 8, and 15, and trastuzumab or pertuzumab was dosed qd×1 on day 1. For all doses, conjugates were dosed intravenously; trastuzumab and pertuzumab were dosed intraperitoneally; all doses of conjugates are reported as antibody / payload; n=10 for each group).

[0272] FIG. 3 provides tumor volume results for SNU-5 tumor-bearing mice treated with vehicle, conjugate 1, conjugate 2, trastuzumab, pertuzumab, a combination of trastuzumab and pertuzumab, a combination of conjugate 1 and trastuzumab, a combination of conjugate 1 and pertuzumab, and a combination of conjugate 1, trastuzumab, and pertuzumab at various dose levels and dosing regimens.

[0273] Example 8: Tumor growth response to administration of HER2 STING agonist antibody-drug conjugates in combination with Enhertu in JIMT-1 Female CB.17 SCID mice were inoculated with JIMT-1 tumor cells (10 × 10 6 The tumor volume was 60–100 mm 3 (Average 76.8mm 3 Animals were randomized into treatment groups when the dose was 1.0 / 0.037mg / kg / group: vehicle, conjugate 2 (1.0 / 0.037mg / kg), conjugate 1 (1.0 / 0.043mg / kg), Enhertu (1.0 / 0.026mg / kg, 3.0 / 0.078mg / kg, or 10.0 / 0.261mg / kg), a combination of conjugate 1 (1.0 / 0.043mg / kg) and Enhertu (1.0 / 0.026mg / kg), conjugate Combinations of conjugate 1 (1.00 / 0.043 mg / kg) with Enhertu (3.0 / 0.078 mg / kg) and combinations of conjugate 1 (1.00 / 0.043 mg / kg) with Enhertu (10.0 / 0.26 mg / kg) were dosed, with conjugate dosed qd x 1 intravenously on day 1 and Enhertu dosed qwk x 2 intravenously on days 1 and 8. Additionally, conjugate 1 (0.30 / 0.013 mg / kg), combination of conjugate 1 (0.3 / 0.13 mg / kg) with Enhertu (1.0 / 0.026 mg / kg), combination of conjugate 1 (0.3 / 0.13 mg / kg) with Enhertu (3.0 / 0.078 mg / kg), and combination of conjugate 1 (0.3 / 0.13 mg / kg) with Enhertu (10.0 / 0.261 mg / kg) were dosed intravenously qwk×2 on days 1 and 8; all doses of conjugates are listed as antibody / payload; n=10 for each group.

[0274] FIG. 4 provides the results of tumor volume in JIMT-1 tumor-bearing mice treated with vehicle, conjugate 1, conjugate 2, Enhertu, and a combination of conjugate 1 and Enhertu at various dose levels and dosing regimens.

[0275] Example 9: Tumor growth response to administration of HER2 STING agonist antibody-drug conjugates in combination with anti-PD-1 in rat HER2 EMT6-RHER2-MSA mammary adenocarcinoma PD-L1 was upregulated in SKOV3 tumors following treatment with conjugate 1 ( FIG. 7 ), suggesting that combining conjugate 1 with immunotherapeutic agents such as PD-1 checkpoint inhibitors may be effective. PD-L1 was observed to be upregulated in mouse and human cells following treatment with conjugate 1.

[0276] Female BALB / c mice were cultured with EMT6-RHER2_MSA (a modified EMT6 cell line overexpressing rat epidermal growth factor receptor 2 (RHER2)) (5 ​​× 10 6 The tumor volume was 60–100 mm 3 (Average=95~109mm 3Animals were randomized into treatment groups when the dose reached 100 / group. Vehicle, conjugate 3 (0.30 / 0.013 mg / kg, 1.0 / 0.04 mg / kg, or 3.0 / 0.12 mg / kg), conjugate 4 (0.30 / 0.012 mg / kg, 1.0 / 0.035 mg / kg, or 3.0 / 0.104 mg / kg) were all dosed qd×1 on day 1. Anti-PD-1 RMP1-14 (1 mg / kg or 5 mg / kg) was dosed biw×2 on days 1, 4, 8, and 11. For the combinations of conjugate 3 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), conjugate 3 (1 / 0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), conjugate 4 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), and conjugate 4 (1.0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), the conjugates were dosed qd×1 on day 1 and anti-PD-1 RMP1-14 was dosed biw×2 on days 1, 4, 8, and 11. For all doses, conjugates were dosed intravenously; anti-PD-1 RMP1-14 was dosed intraperitoneally; all doses of conjugates are reported as antibody / payload; n=10 per group.

[0277] FIG. 5 provides tumor volume results for EMT6-RHER2_MSA tumor-bearing mice treated with vehicle, conjugate 3, conjugate 4, anti-PD-1 RMP1-14, a combination of conjugate 3 and anti-PD-1 RMP1-14, and a combination of conjugate 4 and anti-PD-1 RMP1-14 at various dose levels and dosing regimens. Treatment with conjugate 3 (1.0 / 0.04 mg / kg) resulted in 5 CRs and 5 TFSs. Treatment with conjugate 3 (3.0 / 0.12 mg / kg) resulted in 7 CRs and 7 TFSs. Treatment with conjugate 4 (0.3 / 0.012 mg / kg) resulted in 5 CRs and 5 TFSs. Treatment with conjugate 4 (1.0 / 0.035 mg / kg) resulted in 10 CRs and 10 TFSs. Treatment with conjugate 4 (3.0 / 0.10 mg / kg) resulted in 9 CRs and 9 TFS. Treatment with conjugate 3 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg) resulted in 1 PR. Treatment with conjugate 3 (1.0 / 0.040 mg / kg) and anti-PD-1 RMP1-14 (5 mg) resulted in 8 CRs and 8 TFS. Treatment with conjugate 4 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg) resulted in 1 PR, 8 CRs, and 8 TFS. Treatment with conjugate 4 (1.0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg) resulted in 10 CRs and 10 TFS.

[0278] Example 10: Rechallenge study of tumor growth response following administration of HER2 STING agonist antibody-drug conjugates in combination with anti-PD-1 Tumor-free mice (i.e., mice with complete tumor regression at the end of the study) from Example 9 that had previously been inoculated subcutaneously in the right flank with EMT6-RHER2_MSA and treated with a combination of conjugate 4 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg) or conjugate 4 (0.3 / 0.012 mg / kg) were inoculated with EMT-6-MSA breast adenocarcinoma cells (5×10 per mouse) in the left flank of age-matched untreated animals. 6 cells) subcutaneously and CT26 colon / colorectal cancer cells (3 × 10 per mouse) were inoculated into the right flank. 5 Tumor growth was monitored and measured twice weekly on both flanks, and animals were weighed twice weekly.

[0279] Figures 6A and 6B show tumor volumes in mice previously treated with conjugate 4 (0.3 / 0.012 mg / kg) when rechallenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. Figures 6C and 6D show tumor volumes in mice previously treated with conjugate 4 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg) when rechallenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. Mice previously treated with conjugate 4 or conjugate 4 and anti-PD-1 RMP1-14 (5 mg / kg) experienced rejection of 2 out of 5 and 4 out of 4 tumors, respectively, when rechallenged with EMT-6-MSA cells. Mice previously treated with conjugate 4 or conjugate 4 plus anti-PD-1 RMP1-14 did not reject any cells when rechallenged with CT26 cells. Mice treated with conjugate 4 as monotherapy or in combination with anti-PD-1 RMP1-14 demonstrated immune memory.

[0280] Equivalent The details of one or more aspects of the present disclosure are described in the accompanying description above. Although any method and material similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, currently preferred methods and materials are described. Other features, goals, and advantages of the present disclosure will be clear from the description and claims. In this specification and the appended claims, singular forms include plural referents unless otherwise clearly defined by the context. 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 and publications cited herein are incorporated by reference.

[0281] The foregoing description has been presented for purposes of illustration only, and is not intended to limit the disclosure to the precise form disclosed, but rather to be limited by the claims appended hereto.

Claims

1. A pharmaceutical product comprising at least one HER2-targeted STING agonist antibody-drug conjugate, wherein the at least one HER2-targeted STING agonist antibody-drug conjugate is used in combination with at least one HER2-targeted therapy agent or at least one immunotherapy agent. The HER2-targeted STING agonist antibody-drug conjugate has formula (A): It is a conjugate of, In the formula, the conjugate is Variable heavy chain complementarity determination region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); amino acid sequence Variable heavy chain complementarity determination region 2 (CDRH2) containing; Variable heavy chain complementarity determination region 3 (CDRH3) containing amino acid sequence GGHGYFDL (SEQ ID NO: 7); and amino acid sequence Variable light chain complementarity determination region 1 (CDRL1) containing the amino acid sequence GASSRAT (SEQ ID NO: 13); Variable light chain complementarity determination region 2 (CDRL2) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14); and Variable light chain complementarity determination region 3 (CDRL3) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14). It contains a HER2 antibody that includes d 15 It is approximately 8. The aforementioned pharmaceutical.

2. Use of at least one HER2-targeted STING agonist antibody-drug conjugate in the manufacture of a medicament for treating, preventing, delaying the progression of, or otherwise improving the symptoms of cancer, wherein the HER2-targeted STING agonist antibody-drug conjugate is administered in combination with at least one HER2-targeted therapeutic agent or at least one immunotherapeutic agent. The HER2-targeted STING agonist antibody-drug conjugate has formula (A): It is a conjugate of, In the formula, the conjugate is Variable heavy chain complementarity determination region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); amino acid sequence Variable heavy chain complementarity determination region 2 (CDRH2) containing; Variable heavy chain complementarity determination region 3 (CDRH3) containing amino acid sequence GGHGYFDL (SEQ ID NO: 7); and amino acid sequence Variable light chain complementarity determination region 1 (CDRL1) containing the amino acid sequence GASSRAT (SEQ ID NO: 13); Variable light chain complementarity determination region 2 (CDRL2) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14); and Variable light chain complementarity determination region 3 (CDRL3) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14). It contains a HER2 antibody that includes and has a d15 of approximately 8. The aforementioned use.

3. Use of at least one HER2-targeted therapeutic agent or at least one immunotherapeutic agent in the manufacture of a medicament for treating, preventing, delaying the progression of, or otherwise improving the symptoms of cancer, wherein the at least one HER2-targeted therapeutic agent or at least one immunotherapeutic agent is administered in combination with at least one HER2-targeted STING agonist antibody-drug conjugate. The HER2-targeted STING agonist antibody-drug conjugate has formula (A): It is a conjugate of, In the formula, the conjugate is Variable heavy chain complementarity determination region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); amino acid sequence Variable heavy chain complementarity determination region 2 (CDRH2) containing; Variable heavy chain complementarity determination region 3 (CDRH3) containing amino acid sequence GGHGYFDL (SEQ ID NO: 7); and amino acid sequence Variable light chain complementarity determination region 1 (CDRL1) containing the amino acid sequence GASSRAT (SEQ ID NO: 13); Variable light chain complementarity determination region 2 (CDRL2) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14); and Variable light chain complementarity determination region 3 (CDRL3) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14). It contains a HER2 antibody that includes and has a d15 of approximately 8. The aforementioned use.

4. The use of at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent or at least one immunotherapy agent in the manufacture of a medicament for treating, preventing, delaying the progression of, or otherwise improving the symptoms of cancer, The HER2-targeted STING agonist antibody-drug conjugate has formula (A): It is a conjugate of, In the formula, the conjugate is Variable heavy chain complementarity determination region 1 (CDRH1) containing the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); amino acid sequence Variable heavy chain complementarity determination region 2 (CDRH2) containing; Variable heavy chain complementarity determination region 3 (CDRH3) containing amino acid sequence GGHGYFDL (SEQ ID NO: 7); and amino acid sequence Variable light chain complementarity determination region 1 (CDRL1) containing the amino acid sequence GASSRAT (SEQ ID NO: 13); Variable light chain complementarity determination region 2 (CDRL2) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14); and Variable light chain complementarity determination region 3 (CDRL3) containing the amino acid sequence QQYHHSPLT (SEQ ID NO: 14). It contains a HER2 antibody that includes and has a d15 of approximately 8. The aforementioned use.

5. The aforementioned HER2 antibody, Human HER2 receptor epitopes including residues 452–531 of the extracellular domain of the human HER2 receptor, residues 474–553 of SEQ ID NO: 1, or residues 452–531 of SEQ ID NO:

16. The pharmaceutical product according to claim 1, or the use according to any one of claims 2 to 4, which specifically binds to the target.

6. The pharmaceutical product according to claim 1, comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one HER2-targeted therapeutic agent.

7. The use according to any one of claims 2 to 4, wherein at least one HER2-targeted STING agonist antibody-drug conjugate is administered in combination with at least one HER2-targeted therapeutic agent.

8. The pharmaceutical product according to claim 1, comprising at least one HER2-targeted STING agonist antibody-drug conjugate and at least one immunotherapy agent.

9. The use according to any one of claims 2 to 4, wherein at least one HER2-targeted STING agonist antibody-drug conjugate is administered in combination with at least one immunotherapy agent.

10. The pharmaceutically active ingredient according to claim 1 or the use according to any one of claims 2 to 4, wherein the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of the HER2-targeted therapeutic agent or the immunotherapy agent.

11. The pharmaceutical product according to claim 6, wherein the HER2-targeted therapeutic agent is an antibody or antigen-binding fragment thereof that specifically binds to HER2, a HER2-targeted antibody-drug conjugate that specifically binds to HER2, or a small molecule inhibitor of HER2.

12. The use according to claim 7, wherein the HER2-targeted therapeutic agent is an antibody or antigen-binding fragment thereof that specifically binds to HER2, a HER2-targeted antibody-drug conjugate that specifically binds to HER2, or a small molecule inhibitor of HER2.

13. The pharmaceutical product according to claim 11, wherein the antibody or antigen-binding fragment that specifically binds to HER2 is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination thereof.

14. The use according to claim 12, wherein the antibody or antigen-binding fragment that specifically binds to HER2 is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination thereof.

15. The pharmaceutical product according to claim 13, wherein the HER2 antibody or the HER2 dimerization inhibitor antibody is trastuzumab, pertuzumab, or margetuximab, or a biosimilar thereof.

16. The use according to claim 14, wherein the HER2 antibody or the HER2 dimerization inhibitor antibody is trastuzumab, pertuzumab, or margetuximab, or a biosimilar thereof.

17. The pharmacopoeia according to claim 11, wherein the HER2-targeted therapeutic agent is trastuzumab, pertuzumab, a combination of trastuzumab and pertuzumab, margetuximab, a biosimilar of trastuzumab, a biosimilar of pertuzumab, a combination of a biosimilar of trastuzumab and a biosimilar of pertuzumab, or a biosimilar of margetuximab.

18. The use according to claim 12, wherein the HER2-targeted therapeutic agent is trastuzumab, pertuzumab, a combination of trastuzumab and pertuzumab, margetuximab, a biosimilar of trastuzumab, a biosimilar of pertuzumab, a combination of a biosimilar of trastuzumab and a biosimilar of pertuzumab, or a biosimilar of margetuximab.

19. The pharmaceutical product according to claim 11, wherein the HER2-targeted antibody-drug conjugate that specifically binds to HER2 is ad-trastuzumab emtansine (T-DM1) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan).

20. The use according to claim 12, wherein the HER2-targeted antibody-drug conjugate that specifically binds to HER2 is ad-trastuzumab emtansine (T-DM1) or fam-trastuzumab deruxtecan (trastuzumab deruxtecan).

21. The pharmaceutical product according to claim 11, wherein the HER2 small molecule inhibitor is tucatinib, neratinib, or lapatinib.

22. The use according to claim 12, wherein the HER2 small molecule inhibitor is tucatinib, neratinib, or lapatinib.

23. The pharmaceutical product according to claim 8, wherein the immunotherapy agent is an immune checkpoint inhibitor.

24. The use according to claim 9, wherein the immunotherapy agent is an immune checkpoint inhibitor.

25. The pharmaceutical product according to claim 23, wherein the immune checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof.

26. The use according to claim 24, wherein the immune checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof.

27. The pharmaceutical product according to claim 23, wherein the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

28. The use according to claim 24, wherein the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

29. The pharmaceutical product according to claim 27, wherein the PD-1 inhibitor or the PD-L1 inhibitor is avelumab, durvalumab, dostallimab, pembrolizumab, semiprimab, nivolumab, or atezolizumab.

30. The use according to claim 28, wherein the PD-1 inhibitor or the PD-L1 inhibitor is avelumab, durvalumab, dostallimab, pembrolizumab, semiprimab, nivolumab, or atezolizumab.

31. The use according to any one of claims 2 to 4, wherein the cancer is anal cancer, astrocytoma, leukemia, lymphoma, head and neck cancer, liver cancer, testicular cancer, cervical cancer, sarcoma, hemangioma, esophageal cancer, eye cancer, laryngeal cancer, oral cancer, mesothelioma, skin cancer, myeloma, oral cancer, rectal cancer, colorectal cancer, pharyngeal cancer, bladder cancer, breast cancer, urothelial carcinoma, uterine cancer, ovarian cancer, prostate cancer, lung cancer, non-small cell lung cancer (NSCLC), colon cancer, pancreatic cancer, kidney cancer, gastric cancer, or gastric esophagogastric junction cancer.

32. The use according to claim 31, wherein the cancer is breast cancer, gastric cancer, gastroesophageal junction cancer, colorectal cancer, or non-small cell lung cancer.

33. i) The HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapeutic agent or the immunotherapy agent are administered simultaneously. ii) The HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapeutic agent or the immunotherapy agent are administered sequentially or alternately in either order. iii) The HER2-targeted STING agonist antibody-drug conjugate is administered before the HER2-targeted therapeutic agent or the immunotherapy agent. iv) The HER2-targeted STING agonist antibody-drug conjugate is administered after the HER2-targeted therapeutic agent or the immunotherapy agent. The use described in any one of claims 2 to 4.

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