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

Abstract

The present disclosure provides a combination comprising a STING agonist antibody-drug conjugate that targets HER2 and a HER2 targeting therapy or immunotherapy. The present disclosure also provides the use of the combination in therapy, such as the treatment of cancer.

Description

[0001] Related Applications

[0002] This application claims priority to and the benefit of U.S. Provisional Application No. 63 / 317,472, filed on March 7, 2022, and U.S. Provisional Application No. 63 / 329,680, filed on April 11, 2022. The contents of each of these applications are hereby incorporated by reference in their entirety.

[0003] Electronic Sequence Listing Reference

[0004] The contents of the electronic sequence listing (MRSN_037_001WO_SeqList_ST26.xml; size: 18,657 bytes; and creation date: March 1, 2023) are hereby incorporated by reference in their entirety. Background Art

[0005] Stimulator of interferon genes (STING) is a receptor in the endoplasmic reticulum that transmits innate immune perception of cytosolic pathogen-derived DNA and self-DNA. STING is a 378-amino acid protein that mainly contains three domains: (i) N-terminal transmembrane domain (aa 1-154); (ii) central globular domain (aa 155-341); and (iii) C-terminal tail (aa 342-379). STING can combine with its ligand in a V-shaped conformation to form a symmetrical dimer, but does not completely cover the bound ligand. STING agonists can bind to the pocket region of STING. However, under some severe disease conditions, the STING activation process is easily inhibited, resulting in inactivation of the STING pathway. Therefore, screening and designing effective STING agonists are of great significance for cancer immunotherapy and other infectious disease treatments (including but not limited to obesity, liver injury, lipid metabolism and viral infection). Specific targeting of immune pathways presents opportunities for cancer treatment and may provide higher specificity than cell population-based treatments.

[0006] Antibody-drug conjugates (ADCs) consist of drugs (e.g., small molecules) covalently linked to antibodies. Antibodies represent a targeting mechanism that is tuned to a specific site of action. Upon reaching the site, ADCs are designed to release small molecules (i.e., drugs) that can function in a targeted manner, rather than diffuse throughout the subject's body. This targeted approach allows treatment with drugs that would otherwise require doses that are too high to be toxic when administered systemically, and minimizes the likelihood of hitting non-tumor toxicity.

[0007] A key feature of the innate immune system is the recognition and elimination of foreign agents. Recognition of these pathogenic invaders is achieved through host recognition of evolutionarily conserved microbial structures called pathogen-associated molecular patterns (PAMPs). Host recognition can be achieved through a variety of pathways, such as activation of pattern recognition receptors (PRRs), which ultimately lead to downstream signaling events and culminate in the development of an immune response.

[0008] The antibody-drug conjugates disclosed herein modulate the activity of STING and thus can provide a beneficial therapeutic impact in treating diseases, pathologies and / or conditions in which modulation of STING (stimulator of interferon genes) is beneficial, including but not limited to inflammation, allergic and autoimmune diseases, infectious diseases, cancer, precancerous syndromes, and as vaccine adjuvants. In addition, in combination therapies using two or more drugs in a certain dosing regimen or administration form, the additive or synergistic effects of the biological activities of the two or more drugs can be exploited to enhance efficacy. There is still a need for new immunotherapies to treat diseases, particularly cancer. Summary of the invention

[0009] In some aspects, the present disclosure provides, inter alia, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immunotherapy (e.g., an immune checkpoint inhibitor), wherein the conjugate comprises an antibody or an antigen-binding fragment thereof that specifically binds to a human HER2 receptor epitope.

[0010] In some aspects, the HER2-targeting STING agonist antibody-drug conjugate is a conjugate of structural formula (A):

[0011]

[0012] wherein the conjugate comprises a HER2 antibody comprising a variable heavy chain complementary determining region 1 (CDRH1) comprising the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); a variable heavy chain complementary determining region 2 (CDRH2) comprising the amino acid sequence YISSSSSTIYYADSVKG (SEQ ID NO: 6); a variable heavy chain complementary determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 7); and a variable light chain complementary determining region 1 (CDRL1) comprising the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 12); a variable light chain complementary determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 13); and a variable light chain complementary determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 14), and d 15 About 8.

[0013] In some aspects, a HER2 antibody or antigen-binding fragment thereof that specifically binds to an epitope of a human HER2 receptor comprises residues 452 to 531 of the extracellular domain of a human HER2 receptor, residues 474 to 553 of SEQ ID NO: 1, or residues 452 to 531 of SEQ ID NO:16.

[0014] In some aspects, the present disclosure provides compositions comprising combination therapies including at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immunotherapy (e.g., an immune checkpoint inhibitor).

[0015] In some aspects, the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of HER2-targeted therapy or immunotherapy (eg, immune checkpoint inhibitors).

[0016] In some aspects, the present disclosure provides a composition comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2-targeting therapy.

[0017] In some aspects, the present disclosure provides a composition comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one immunotherapy (e.g., an immune checkpoint inhibitor).

[0018] In some aspects, the HER2 targeted therapy 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 HER2 small molecule inhibitor.

[0019] In some aspects, the present disclosure provides a composition comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one antibody or antigen-binding fragment thereof that specifically binds to HER2, at least one STING agonist antibody-drug conjugate targeting HER2 that specifically binds to HER2, or at least one HER2 small molecule inhibitor.

[0020] In some aspects, the HER2 targeted therapy is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody.

[0021] In some aspects, the HER2-targeted therapy is trastuzumab, pertuzumab, a combination thereof, or magetuximab or a biosimilar thereof.

[0022] In some aspects, the HER2 targeted therapy is trastuzumab, pertuzumab, or a combination thereof.

[0023] In some aspects, the HER2-targeted therapy is magetuximab or a biosimilar thereof.

[0024] In some aspects, the HER2-targeted therapy is a HER2-targeted antibody-drug conjugate that specifically binds to HER2, such as, for example, trastuzumab emtansine (T-DM1). or trastuzumab delutec (trastuzumab-delutec)

[0025] In some aspects, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 and trastuzumab emtansine (T-DM1). or trastuzumab delutec (trastuzumab-delutec) combination.

[0026] In some aspects, the HER2 targeted therapy is a HER2 small molecule inhibitor, such as, for example, tucatinib, neratinib, or lapatinib.

[0027] In some aspects, the combination includes at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with tucatinib, neratinib, or lapatinib.

[0028] In some aspects, a combination comprising a HER2-targeting 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-targeting STING agonist antibody-drug conjugate that specifically binds to HER2, or at least one HER2 small molecule inhibitor.

[0029] 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.

[0030] In some aspects, immune checkpoint inhibitors suitable for the combinations and methods of the present disclosure are PD-1 inhibitors or PD-L1 inhibitors.

[0031] In some aspects, the combination includes a HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with a PD-1 inhibitor or a PD-L1 inhibitor.

[0032] In some aspects, the combination includes a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein and an immune checkpoint inhibitor, such as, for example, avelumab, durvalumab, dotalimumab, pembrolizumab, cemiplimab, nivolumab or atezolizumab. In some aspects, the combination includes a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein and dotalimumab. In some aspects, the combination includes a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein and pembrolizumab.

[0033] In some aspects, a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein is administered in combination with an immune checkpoint inhibitor (e.g., such as avelumab, durvalumab, dotalimumab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab). In some aspects, a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein is administered in combination with dotalimumab. In some aspects, a combination of a STING agonist antibody-drug conjugate targeting HER2 disclosed herein is administered in combination with pembrolizumab.

[0034] The combination of a STING agonist antibody-drug conjugate targeting HER2 and a HER2 targeted therapy or an immune checkpoint inhibitor can be used to treat a condition in a subject, such as, for example, cancer. For example, the combination disclosed herein comprising a STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor can be used to treat, prevent, delay cancer progression in a subject, or otherwise improve cancer symptoms.

[0035] In some embodiments, the cancer is, for example, selected from 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, 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, stomach cancer and gastroesophageal junction cancer.

[0036] In some aspects, the combination therapies disclosed herein can be used to treat, prevent, slow the progression of, or otherwise improve breast cancer symptoms, gastric cancer symptoms, gastroesophageal junction cancer symptoms, non-small cell lung cancer (NSCLC) symptoms, or colorectal cancer symptoms in a subject.

[0037] In some aspects, the combination therapies disclosed herein can be used 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 a subject.

[0038] In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying breast cancer progression or otherwise improving breast cancer symptoms in a subject. In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying HER2+ breast cancer progression or otherwise improving HER2+ breast cancer symptoms in a subject. In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying HER2- breast cancer progression or otherwise improving HER2- breast cancer symptoms in a subject. In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying metastatic HER2+ breast cancer progression or otherwise improving metastatic HER2+ breast cancer symptoms in a subject. In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying metastatic HER2+ breast cancer progression or otherwise improving metastatic HER2+ breast cancer symptoms in a subject who has received at least one, at least two, at least three or at least four prior breast cancer treatments, treating, preventing, delaying metastatic HER2+ breast cancer progression or otherwise improving metastatic HER2+ breast cancer symptoms. In some aspects, the combination therapy disclosed herein can be used for treating, preventing, delaying metastatic HER2+ breast cancer progression or otherwise improving metastatic HER2+ breast cancer symptoms in a subject who has received three or more prior breast cancer treatments.

[0039] In some aspects, the combination of a STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor used in any aspect of the methods and uses provided herein can be administered at any stage of the disease. For example, such combination therapy can be administered to cancer patients at any stage from early stage to metastatic.

[0040] The combination therapy including a STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor used in any aspect 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 toxin 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 a standard first-line chemotherapy agent, such as, for example, trastuzumab emtansine (Hercel), 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, ixabepilone, doxorubicin, fluorouracil, oxaliplatin, fluoropyrimidine, irinotecan, ramucirumab, mitomycin, folinic acid, cetuximab, bevacizumab, erlotinib, afatinib, crizotinib, pemetrexed, ceritinib, etoposide, vinblastine, vincristine, ifosfamide, liposomal doxorubicin, topotecan, hexamethylmelamine, melphalan or leuprolide acetate. In some aspects, the additional agent is Herceptin (enmetuzumab).

[0041] In some aspects, a combination of a STING agonist antibody-drug conjugate targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor and an additional agent is formulated as a single therapeutic composition, and the components are administered simultaneously. Alternatively, the STING agonist antibody-drug conjugate targeting HER2, the HER2 targeted therapy or immune checkpoint inhibitor, and (if any) additional agents are separated from each other, for example, each is formulated into a separate therapeutic composition, and can be administered simultaneously, or at different times during the treatment regimen. For example, a STING agonist antibody-drug targeting HER2 is administered before administering a HER2 targeted therapy or immune checkpoint inhibitor combination; a STING agonist antibody-drug targeting HER2 is administered after administering a HER2 targeted therapy or immune checkpoint inhibitor combination. As described herein, a STING agonist antibody drug targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor combination can be administered in a single dose or multiple doses.

[0042] The pharmaceutical compositions according to the present disclosure may include a suitable carrier. These pharmaceutical compositions may be included in a kit, such as, for example, a diagnostic kit.

[0043] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those of ordinary skill in the art to which the present disclosure belongs. In the specification, unless the context clearly provides otherwise, the singular also includes the plural. Although methods and materials similar or equivalent to the methods and materials 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. The references cited herein are not considered to be prior art of the claimed invention. In the event of a conflict, the present specification including definitions will prevail. In addition, materials, methods and embodiments are illustrative only and are not intended to be limiting. In the event of a conflict between the chemical structure and the name of the compound disclosed herein, the chemical structure will prevail.

[0044] Other features and advantages of the disclosure will be apparent from the following detailed description, and from the claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The graph shows the anti-tumor efficacy of Conjugate 1, trastuzumab, the combination of Conjugate 2 and trastuzumab, and the combination of Conjugate 1 and trastuzumab at different dose levels and dosing schedules in mice bearing SKOV3 tumors.

[0047] Figure 2 The graph shows the anti-tumor efficacy of Conjugate 1; the combination of Conjugate 1 and Trastuzumab; the combination of Conjugate 1 and Pertuzumab; the combination of Conjugate 1, Trastuzumab and Pertuzumab; the combination of Conjugate 2 and Trastuzumab; the combination of Conjugate 2 and Pertuzumab, and the combination of Conjugate 2, Trastuzumab and Pertuzumab in mice bearing JIMT-1 tumors at different dose levels and dosing schedules.

[0048] Figure 3 The graph shows the anti-tumor efficacy of Conjugate 1; Conjugate 2; Trastuzumab; Pertuzumab; the combination of Trastuzumab and Pertuzumab; the combination of Conjugate 1 and Trastuzumab; the combination of Conjugate 1 and Pertuzumab; and the combination of Conjugate 1, Trastuzumab, and Pertuzumab in mice bearing SNU-5 tumors at different dose levels and dosing schedules.

[0049] Figure 4 The graph shows the anti-tumor efficacy of Conjugate 1, Conjugate 2, Enhertu, and the combination of Conjugate 1 and Enhertu at different dose levels and dosing schedules in mice bearing JIMT-1 tumors.

[0050] Figure 5The figure shows 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 mice bearing EMT6-RHER2_MSA tumors at different dose levels and dosing schedules.

[0051] Fig. 6A and 6B The graphs show tumor volumes of mice previously treated with conjugate 4 when re-challenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. Figure 6C and 6D The graph shows tumor volume of mice previously treated with conjugate 4 and anti-PD-1 RMP1-14 when re-challenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively.

[0052] Figure 7 ' is a series of graphs depicting PD-L1 expression in murine and human SKOV3 tumors treated with vehicle, control ADC, and Conjugate 1 (a STING agonist ADC targeting HER2). DETAILED DESCRIPTION OF THE INVENTION

[0054] The present disclosure provides novel HER2-targeting STING agonist antibody-drug conjugates, pharmaceutical compositions containing the conjugates, and various uses of the conjugates.

[0055] definition

[0056] The chemical names provided for the intermediate compounds and / or compounds described in the present disclosure may refer to any one of the tautomeric representatives of such compounds (in some cases, such alternative names are provided along with the experimental names). It should be understood that any reference to a named compound (an intermediate compound or compound of the present disclosure) or a structurally described 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.

[0057] It should be understood that the terms "in some aspects," "in some aspects of the disclosure," and "some aspects of the compounds of the disclosure" can be used interchangeably where appropriate.

[0058] The terms "about", "approximately" or "approximately" when used in conjunction with a numerical value are intended to include a set or range of values. In some aspects, "about X" includes a range of values ​​of ±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% greater or less than a specified value. In some aspects, the term "about" refers to a range of values ​​that are 2% greater or less than a specified value. In some aspects, the term "about" refers to a range of values ​​that are 1% greater or less than a specified value.

[0059] Unless otherwise specified herein, the enumeration of a range of values ​​is intended only as a shorthand method of individually referring to each individual value falling within the range, and each individual value is incorporated into the specification as if it were individually listed herein. Unless otherwise specified, ranges used herein include both limits of the range. In some aspects, the expressions "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 "between X and Y" and "the range from X to Y" include X and Y and integers therebetween.

[0060] As used herein, the term "HER2" (also referred to as ErbB-2, NEU, HER-2, and CD340) refers to human epidermal growth factor receptor-2 (SwissProt P04626) when used herein, and includes any variants, isomers, and species homologs of HER2, which are naturally expressed by cells (including tumor cells) or expressed on cells transfected with the HER2 gene. Species homologs include rhesus HER2 (Macaca mulatta; Genbank accession number GI: 109114897). These terms are synonymous and can be used interchangeably.

[0061] As used herein, the term "HER2 antibody" or "anti-HER2 antibody" is an antibody that specifically binds to the antigen HER2.

[0062] The term "antibody" as used herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) and antibody fragments, as 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)).

[0063] The term "antibody fragment" refers to a molecule other than an intact antibody that contains a portion of an 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.

[0064] As used herein, the term "an 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 competition assay, and conversely, the reference antibody blocks the binding of the antibody to its antigen by 50% or more in a competition assay. Exemplary competition assays are provided herein.

[0065] When used in the context of two or more antibodies herein, the term "competes with" or "cross-competes with" indicates that two or more antibodies compete for binding to HER2; if one antibody competes with one or more other antibodies by 25% or more, the antibody "blocks" or "cross-blocks" one or more other antibodies from binding to HER2, wherein 25%-74% represents "partial blocking" and 75%-400% represents "complete blocking". Unless otherwise defined or negated by the context, the terms "compete with", "cross-compete with", "blocking" or "cross-blocking" are also intended to cover such antibody pairs when used herein.

[0066] The term "epitope" refers to a specific site on an antigen molecule to which an antibody binds.

[0067] As used herein, the term "independently" means that when more than one substituent is selected from a plurality of possible substituents, the substituents may be the same or different.

[0068] As used herein, the term "pharmaceutically acceptable" refers to compounds, conjugates, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problems or complications, and with a reasonable benefit / risk ratio commensurate.

[0069] As used herein, the term "pharmaceutical composition" refers to a mixture, preparation or solution, which contains at least one therapeutic agent for administration to a subject (e.g., a mammal or a human) to treat a specific disease or condition affecting the subject. The drug combination of the present invention can be formulated into a pharmaceutical composition suitable for enteral or parenteral administration, such as a sugar-coated tablet, tablet, capsule or suppository or ampoule. If not otherwise specified, these are prepared in a manner known per se, such as by various conventional mixing, crushing, direct compression, granulation, sugar coating, dissolution, freeze-drying processes or manufacturing techniques that are obvious to those skilled in the art. It should be understood that the unit content of the combination partner contained in a single dose of each dosage form does not necessarily constitute an effective amount itself, because the necessary effective amount can be achieved by administering multiple dosage units. A person of ordinary skill in the art can select one or more of the above-mentioned carriers for the specific desired characteristics of the dosage form by routine experiments, without any excessive burden. The amount of each carrier used can vary within the conventional range in the art. The pharmaceutical composition provided herein can be in the form of a sterile injection preparation, such as a sterile injection aqueous or oily suspension. The suspension can be prepared according to known techniques using those suitable dispersants or wetting agents and suspending agents mentioned above. Aseptic injection preparation can also be a sterile injection solution or suspension in a nontoxic parenteral acceptable diluent or solvent, such as a solution in 1,3-butanediol or prepared as a lyophilized powder. Available acceptable carriers and solvents include water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils can usually be used as solvents or suspension media. For this purpose, any gentle fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids (such as oleic acid) can be used to prepare injections equally.

[0070] As used herein, the term "treating" or "treatment" describes the management and care of a patient to combat a disease, disorder, or condition, and includes the administration of a compound of the present disclosure, or a pharmaceutically acceptable salt, polymorph, or solvate thereof, to alleviate the symptoms or complications of the disease, disorder, or condition, or to eliminate the disease, disorder, or condition. The term "treatment" may also include treatment of in vitro cells or animal models.

[0071] As used herein, the terms "preventing," "prevent," or "protecting against" describe the reduction or elimination of the onset of symptoms or complications of such disease, disorder or condition.

[0072] As used herein, the term "subject" includes human and non-human animals, as well as cell lines, cell cultures, tissues and organs. In some embodiments, 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 embodiments, the subject is a human.

[0073] As used herein, the term "subject in need" refers to a subject suffering from a disease or having an increased risk of illness. A subject in need may be a subject previously diagnosed or confirmed to have a disease or lesion disclosed herein. A subject in need may also be a subject suffering from a disease or lesion disclosed herein. Alternatively, a subject in need may be a subject with an increased risk of suffering from such a disease or lesion relative to the general population (i.e., a subject susceptible to such lesions relative to the general population). A subject in need may suffer from a refractory or resistant disease or lesion disclosed herein (i.e., a disease or lesion disclosed herein that is not responsive or has not yet responded to treatment). The subject may be resistant at the beginning of treatment or may become resistant during treatment. In some embodiments, a subject in need has received all known effective therapies for a disease or lesion disclosed herein and has failed. In some embodiments, a subject in need has received at least one previous therapy.

[0074] The term "therapeutically effective amount" refers to that amount of an active compound or agent, including a conjugate of the present disclosure, which elicits the biological or medicinal response in a tissue system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician, including alleviation or partial alleviation of the symptoms of the disease, syndrome, condition or disorder being treated.

[0075] As defined herein, "effective amount" is intended to mean an amount sufficient to effectively treat or prevent when the conjugate is administered to a patient in need of such treatment. The amount of a given conjugate corresponding to such an amount will depend on factors such as the specific conjugate (e.g., the potency (pICso), efficacy (EC 50 ) and biological half-life), the disease condition and its severity, the identity of the patient in need of treatment (e.g., age, size and weight), but can still be routinely determined by those skilled in the art. Similarly, the duration of conjugate treatment and the time period of administration (the time period between doses and the timing of doses, such as before / during / after meals) will vary depending on the identity of the mammal in need of treatment (e.g., weight), the specific conjugate and its properties (e.g., pharmacokinetic properties), the disease or pathology and its severity, and the specific composition and method used, but can still be determined by those skilled in the art.

[0076] The term "composition" refers to a product comprising the specified ingredients in therapeutically effective amounts, as well as any product which results, directly or indirectly, from combination of the specified ingredients in the specified amounts.

[0077] As used herein, the term "pharmaceutically acceptable excipient" means an excipient that can be used to prepare a pharmaceutical composition that is substantially safe, non-toxic, and has neither biological nor other adverse effects, and includes excipients that are acceptable for veterinary use as well as human pharmaceutical use. As used in the specification and claims, "pharmaceutically acceptable excipient" includes one or more than one such excipient.

[0078] As used herein, the term "STING agonist" refers to a compound or moiety that is capable of interacting with STING, for example, by binding to STING and / or inducing downstream signal transduction (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, STING pathway activation results in increased production of type I interferons (primarily IFN-a and IFN-b) and / or expression of interferon-stimulated genes.

[0079] As used herein, the term "STING agonist drug moiety" refers to a moiety derived from a STING agonist and capable of interacting with STING. In some aspects, the STING agonist drug moiety is a moiety derived from a STING agonist to allow the moiety to be linked to the remainder of the conjugate of the present disclosure.

[0080] As used herein, "immunotherapy" refers to an agent that activates or suppresses the immune system or a component of the immune system. Exemplary immunotherapies include, but are not limited to, monoclonal antibodies, immune checkpoint inhibitors, vaccines, cytokine therapy, adoptive cell therapy, or immune system modulators.

[0081] As used herein, "immune checkpoint inhibitors" or "immune checkpoint inhibitory agents" or "immune checkpoint blockers" or "immune checkpoint modulators" refer to agents that bind to inhibitory immune checkpoint proteins and block their activity, thereby enabling the immune system to recognize tumor cells and allow for a sustained immunotherapy response. Inhibition can be competitive or non-competitive inhibition, and can be steric or allosteric. In the case where the immune checkpoint protein is an immunostimulatory protein, the immune checkpoint inhibitor acts to promote the activity of the immunostimulatory protein, such as by binding to and activating the stimulatory immune checkpoint protein or by inhibiting, by interfering with (e.g., by binding to or inactivating) an inhibitor of the stimulatory immune checkpoint protein. An example of an immune checkpoint inhibitor is an anti-immune checkpoint protein antibody.

[0082] As used herein, "immune checkpoints" refer to inhibitory pathways of the immune system that are responsible for maintaining self-tolerance and regulating the duration and amplitude of physiological immune responses in peripheral tissues to minimize collateral tissue damage. Immune checkpoints are regulated by immune checkpoint proteins.

[0083] As used herein, "immune checkpoint protein" refers to a protein that regulates or modulates the extent of an immune response, such as a receptor (e.g., CTLA4 or PD-1) or a ligand (e.g., PD-L1). Immune checkpoint proteins can be inhibitory or stimulatory. Specifically, immune checkpoint proteins are inhibitory to the activation of an immune response. Therefore, inhibition of inhibitory immune checkpoint proteins can stimulate or activate an immune response, such as T cell activation and proliferation.

[0084] As used herein, "combination therapy" refers to a treatment in which two or more therapeutic agents (e.g., at least two or at least three therapeutic agents) are administered to a subject for the treatment of a disease or lesion. For purposes herein, combination therapy includes therapy using a STING agonist antibody-drug conjugate targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor.

[0085] As used herein, "co-administration", "co-administering" or "co-administered" refers to the administration of at least two different therapeutic agents in close enough time. Such administration can be performed in any order, including simultaneous administration, and time intervals ranging from seconds to days. Such administration can also include more than a single administration of one agent and / or independently of another agent. Administration of the agents can be performed by the same or different routes.

[0086] As used herein, the term "simultaneous administration" related to drug administration refers to a drug administration method that allows each drug to be present in the subject's body at the same time. In addition to concomitant administration (via the same or alternative routes) of drugs, simultaneous administration can also include administration (via the same or alternative routes) of drugs at different times.

[0087] As used herein, the terms "at least one" item or "one or more" items each include a single item selected from the list as well as a mixture of two or more items selected from the list.

[0088] As used herein, the term "immune response" relates to any one or more of the following: a specific immune response, a nonspecific immune response, both specific and nonspecific responses, an innate response, a primary immune response, an adaptive immunity, a secondary immune response, a memory immune response, immune cell activation, immune cell proliferation, immune cell differentiation, and cytokine expression.

[0089] The conjugates of the present disclosure can be used in methods for treating or ameliorating viral infections, diseases, syndromes, conditions or lesions affected by STING agonism. Such methods include, consist of and / or consist essentially of: administering a therapeutically effective amount of a conjugate of the present disclosure, or an enantiomer, diastereomer, solvate or pharmaceutically acceptable salt thereof to a subject (including animals, mammals and humans) in need of such treatment, improvement and / or prevention.

[0090] As used herein, the term "conjugate(s) of the disclosure" or "conjugate(s) of the present disclosure" means any form of the conjugate as defined herein, i.e. any tautomeric form, any isomeric form, any salt or non-salt form (e.g. as a free acid or base form, or as a salt, in particular a pharmaceutically acceptable salt thereof) and any physical form thereof (e.g. including non-solid forms (e.g. liquid or semi-solid forms) and solid forms (e.g. amorphous or crystalline forms, specific polymorphic forms, solvate forms, including hydrate forms (e.g. monohydrates, dihydrates and hemihydrates)), as well as mixtures of various forms.

[0091] Thus, the present disclosure includes the conjugates as disclosed herein, in any salt or non-salt form thereof and in any physical form thereof, as well as mixtures of the various forms. While these are included in the present disclosure, it is understood that the conjugates of the present disclosure, in any salt or non-salt form thereof and in any physical form thereof, may have different activity levels, different bioavailabilities and different handling properties for formulation purposes.

[0092] It should be understood that throughout this specification, when a composition is described as having, including or comprising a particular component, it is also envisioned that the composition consists essentially of or consists of said component. Similarly, when a method or process is described as having, including or comprising a particular process step, the process also consists essentially of or consists of said process step. Further, it should be understood that the order of steps or the order in which certain actions are performed is not important as long as the present invention remains feasible. In addition, two or more steps or actions may be performed simultaneously.

[0093] Unless otherwise indicated, all percentages and ratios used herein are calculated by weight. Other features and advantages of the present disclosure are apparent from the various embodiments. The provided embodiments illustrate the different components and methods that can be used to practice the present disclosure. The embodiments do not limit the disclosure claimed. Based on the present disclosure, those skilled in the art can identify and adopt other components and methods that can be used to practice the present disclosure.

[0094] HER2 Antibodies

[0095] In some aspects, the HER2 antibody suitable for conjugation is combined with human HER2 in soluble form or membrane-bound form (i.e., when expressed on the cell surface). In some aspects, the disclosure provides monoclonal antibodies that are combined with HER2 and are humanized or fully human. In some aspects, the disclosure provides monoclonal antibodies that specifically bind HER2. These antibodies are collectively referred to as "HER2" antibodies in this article.

[0096] In some aspects, the HER2 antibody suitable for conjugation has an equilibrium dissociation constant (K) of ≤ 1 uM (eg, ≤ 100 nM; ≤ 10 nM; ≤ 1 nM). d or K D ) binds to the HER2 epitope. 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 exhibit a K d Approximately in the range of ≤1 nM to about 1 pM.

[0097] In some aspects, the HER2 antibodies disclosed herein are used to regulate, 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, regulating PI3K-Akt pathway activity. In some aspects, HER2 antibodies completely or partially inhibit HER2 functional activity by partially or completely regulating, blocking, inhibiting, reducing, antagonizing, neutralizing or otherwise interfering with PI3K-Akt pathway activity. PI3K-Akt pathway activity is assessed using any art-recognized method for detecting PI3K-Akt pathway activity, including but not limited to detecting the level of phosphorylated Akt in the presence and absence of antibodies or antigen-binding fragments disclosed herein.

[0098] In some aspects, a HER2 antibody is considered to fully 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.

[0099] In some aspects, exemplary HER2 antibodies disclosed herein include XMT-1519 antibodies. This antibody is specific for human HER2 and has been shown to inhibit the functional activity of HER2 in vitro.

[0100] In some embodiments, the antibodies or antigen-binding fragments thereof disclosed herein, including HER2 monoclonal antibody XMT-1519, include heavy chains (HC), heavy chain variable regions (VH), light chains (LC), and light chain variable regions (VL), as shown in the amino acids and corresponding nucleic acid sequences shown in Table 1 below. The variable heavy chain region and variable light chain region of each antibody are shaded in the following amino acid sequences. The complementarity determining regions (CDRs) of the heavy and light chains are underlined in the amino acid sequences shown below.

[0101] Table 1: Sequences of HER2 human or humanized monoclonal antibody XMT-1519

[0102]

[0103] 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.

[0104] 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.

[0105] In some embodiments, the antibodies disclosed herein exhibit HER2 binding properties different from those described in the art. In some embodiments, the antibodies disclosed herein bind to different epitopes of HER2 because they cross-block each other's binding to HER2 but do not block the binding of trastuzumab, pertuzumab, Fab37 or chA21 to HER2. Further, in contrast to known antibodies, the antibodies disclosed herein can be effectively internalized into cells expressing HER2 without promoting cell proliferation.

[0106] In some embodiments, the antibodies disclosed herein are fully human monoclonal antibodies that bind to new epitopes and / or have other favorable properties for therapeutic use. In some embodiments, exemplary properties include, but are not limited to, favorable binding properties to cancer cells expressing human HER2 at high or low levels, specific binding to human and rhesus monkey recombinant HER2, effective internalization after binding to HER2, high ability to kill cancer cells expressing HER2 at high or low levels when administered as an antibody drug conjugate (ADC), no significant agonistic effect on the proliferation of cancer cells expressing HER2, and / or providing effective antibody-dependent cell-mediated cytotoxicity (ADCC) to kill cells expressing HER2, and any combination of the above properties.

[0107] 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, wherein the epitope includes residues 452 to 531 of the extracellular domain of the human HER2 receptor, residues 474 to 553 of SEQ ID NO: 1, or residues 452 to 531 of SEQ ID NO: 16.

[0108] 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 human HER2 receptor domain IV, but do not cross-compete with antibodies that bind to human HER2 receptor epitope 4D5. In some embodiments, the antibodies or antigen-binding fragments thereof described herein do not cross-compete with trastuzumab for binding to human HER2 receptors, because trastuzumab is known to bind to human HER2 receptor epitope 4D5. As used herein, the term human HER2 receptor epitope 4D5 refers to amino acid residues 529 to 627 of the extracellular domain of the human HER2 receptor, residues 551 to 649 of SEQ ID NO: 1, or residues 529 to 627 of SEQ ID NO: 16. In some embodiments, the antibody or antigen-binding fragment thereof also binds to at least one epitope on the rhesus monkey HER2 receptor.

[0109] 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 comprising residues 452 to 500 of the extracellular domain of the human HER2 receptor, residues 474 to 522 of SEQ ID NO: 1, or residues 452 to 500 of SEQ ID NO: 16.

[0110] In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that specifically bind to a human HER2 receptor epitope, the epitope comprising at least one amino acid residue selected from the group consisting of amino acid residues E521, L525, and R530 of the extracellular domain of a human HER2 receptor, such as residues 543, 547, and 552 of SEQ ID NO: 1, and residues 521, 525, and 530 of SEQ ID NO: 16. 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 a human HER2 receptor, the epitope comprising at least two amino acid residues selected from the group consisting of amino acid residues E521, L525, and R530 of the extracellular domain of a human HER2 receptor. In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that specifically bind to a human HER2 receptor epitope, the epitope comprising at least amino acid residues E521, L525, and R530 of the extracellular domain of a 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 rhesus HER2 receptor.

[0111] In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that bind to at least a portion of human HER2 receptor domain III and at least a portion of the N-terminus of domain IV, but do not cross-compete with Fab37 monoclonal antibodies or antibodies that bind to human HER2 receptor epitope 4D5. In some embodiments, the antibodies or antigen-binding fragments described herein do not cross-compete with Fab37 monoclonal antibodies and / or trastuzumab for binding to human HER2 receptors. In some embodiments, the antibodies or antigen-binding fragments thereof also bind to at least one epitope on the rhesus monkey HER2 receptor.

[0112] 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 comprising residues 520 to 531 of the extracellular domain of the human HER2 receptor, residues 542 to 553 of SEQ ID NO: 1, or residues 520 to 531 of SEQ ID NO: 16.

[0113] In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that specifically bind to a human HER2 receptor epitope, wherein the epitope comprises 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, such as 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. In some embodiments, antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to human HER2 receptor extracellular domain epitopes, and the epitopes include 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, antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to human HER2 receptor extracellular domain epitopes, and the epitopes include 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 one or all of these antibodies or their antigen-binding fragments also bind to at least one epitope on rhesus monkey HER2 receptor.

[0114] In some embodiments, the antibodies disclosed herein also include antibodies or antigen-binding fragments thereof that specifically bind to an epitope of a human HER2 receptor, the epitope comprising at least one amino acid residue selected from residues C453, H473, N476, R495, H497 and W499 of the extracellular domain of a human HER2 receptor, such as 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. 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 a human HER2 receptor, the epitope comprising 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 a human HER2 receptor. In some embodiments, the antibodies disclosed herein include antibodies or antigen-binding fragments thereof that specifically bind to human HER2 receptor extracellular domain epitopes, the epitopes comprising at least human HER2 receptor extracellular domain amino acid residues C453, H473, N476, R495, H497 and W499. In some embodiments, any one or all of these antibodies or their antigen-binding fragments also bind to at least one epitope on the rhesus monkey HER2 receptor.

[0115] In some embodiments, these antibodies are specific to human HER2, and they have been shown to regulate (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, the rate of internalization of these antibodies from the cell surface of HER2-expressing cells is the same or substantially similar to the rate of internalization of trastuzumab or its biosimilars. In some embodiments, the internalization rate of these antibodies and antigen-binding fragments is about 50% of the total surface binding at time 0 within 4 hours.

[0116] In some embodiments, the antibodies disclosed herein comprise a heavy chain variable region whose amino acid sequence 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 a sequence selected from SEQ ID NO:2, and a light chain variable region whose amino acid sequence 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 a sequence selected from SEQ ID NO:9.

[0117] 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.

[0118] In some embodiments, the antibodies disclosed herein comprise the heavy chain variable region amino acid sequence of SEQ ID NO:2 and the light chain variable region amino acid sequence of SEQ ID NO:9.

[0119] In some embodiments, an 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.

[0120] In some embodiments, 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.

[0121] 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 located in the CDR region, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more conservative substitutions are cumulatively generated across all CDRs. In some embodiments, up to 1, 2, 3 or 4 conservative amino acid substitutions may be present in each CDR sequence, such as in SEQ ID NOs: 5-7 and 12-14.

[0122] Those skilled in the art will recognize that it does not take undue experimentation to determine whether a monoclonal antibody has the same specificity as monoclonal antibody XMT-1519 by determining whether the former prevents the latter from binding to a natural binding partner or other molecule known to be associated with HER2. In some embodiments, if the tested monoclonal antibody competes with a monoclonal antibody disclosed herein, as shown by reduced binding of the monoclonal antibody disclosed herein, then the two monoclonal antibodies bind to the same or closely related epitope.

[0123] In some embodiments, an alternative method to determine whether a monoclonal antibody has the specificity of a monoclonal antibody disclosed herein is to pre-incubate a monoclonal antibody disclosed herein with soluble HER2 (with which it normally reacts), and then add the tested monoclonal antibody to determine whether the ability of the tested monoclonal antibody to bind HER2 is inhibited. If the tested monoclonal antibody is inhibited, it is likely to have an epitope specificity identical to or functionally equivalent to that of a monoclonal antibody disclosed herein.

[0124] In some embodiments, screening of the monoclonal antibodies disclosed herein can also be performed, for example, by measuring HER2-mediated PI3K-Akt pathway activity, and determining whether the test monoclonal antibody is able to regulate, block, inhibit, reduce, antagonize, neutralize or otherwise interfere with PI3K-Akt pathway activity. In some embodiments, HER2 antibodies suitable for conjugation can be generated and purified by well-known techniques, such as WO 2015 / 195917 and PCT / US2018 / 019873, each of which is incorporated herein by reference in its entirety.

[0125] STING agonist antibody conjugate targeting HER2

[0126] The present invention relates to combination therapies comprising immunoconjugates of HER2-targeting antibodies conjugated to STING agonists.

[0127] In some embodiments, the present disclosure specifically provides combination therapies comprising at least one STING agonist antibody-drug conjugate targeting HER2, and at least one HER2 targeted therapy or at least one immunotherapy (e.g., an immune checkpoint inhibitor), wherein the conjugate comprises an antibody or antigen-binding fragment thereof that specifically binds to a human HER2 receptor epitope.

[0128] In some embodiments, the HER2-targeting STING agonist antibody-drug conjugate is a conjugate of formula (A):

[0129]

[0130] wherein the conjugate comprises a HER2 antibody comprising a variable heavy chain complementary determining region 1 (CDRH1) comprising the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); a variable heavy chain complementary determining region 2 (CDRH2) comprising the amino acid sequence YISSSSSTIYYADSVKG (SEQ ID NO: 6); a variable heavy chain complementary determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 7); and a variable light chain complementary determining region 1 (CDRL1) comprising the amino acid sequence RASQSVSSSYLA (SEQ ID NO: 12); a variable light chain complementary determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 13); and a variable light chain complementary determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 14), and d 15 About 8.

[0131] In some embodiments, the HER2 antibody or antigen-binding fragment thereof specifically binds to an epitope of the human HER2 receptor comprising residues 452 to 531 of the extracellular domain of the human HER2 receptor, residues 474 to 553 of SEQ ID NO: 1, or residues 452 to 531 of SEQ ID NO: 16.

[0132] In some embodiments, d 15 is 2, 4, 6 or 8. In some embodiments, d 15 6 or 8.

[0133] In some embodiments, d 15 is 8. In some embodiments, d 15 is 6.

[0134] HER2-targeted therapy

[0135] Any suitable HER2 targeted therapy is contemplated herein for use in the combinations and methods of the present disclosure. In some embodiments, the HER2 targeted therapy is an antibody or antigen-binding fragment thereof that specifically binds to HER2, a second HER2-targeting STING agonist antibody-drug conjugate that specifically binds to HER2, or a HER2 small molecule inhibitor.

[0136] In some embodiments, the HER2 targeted therapy is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination of a HER2 antibody and a HER2 dimerization inhibitor antibody.

[0137] In some embodiments, HER2 targeted therapy is a combination of HER2 antibody, HER2 dimerization inhibitor antibody or HER2 antibody and HER2 dimerization inhibitor antibody. In some embodiments, HER2 antibody, HER2 dimerization inhibitor antibody or HER2 antibody and HER2 dimerization inhibitor antibody combination is trastuzumab, pertuzumab or magetuximab or its biosimilar. In some embodiments, HER2 targeted therapy is trastuzumab or pertuzumab or its combination. In some embodiments, HER2 targeted therapy is trastuzumab. In some embodiments, HER2 targeted therapy is pertuzumab. In some embodiments, HER2 targeted therapy is magetuximab or its biosimilar. In some embodiments, HER2 targeted therapy is trastuzumab biosimilar or pertuzumab biosimilar or trastuzumab biosimilar and pertuzumab biosimilar combination. In some embodiments, HER2 targeted therapy is trastuzumab biosimilar. In some embodiments, HER2 targeted therapy is pertuzumab biosimilar. In some embodiments, the HER2-targeted therapy is a combination of a trastuzumab biosimilar and a pertuzumab biosimilar.

[0138] In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure in combination with the following: 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 includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with the following: trastuzumab or pertuzumab or a combination thereof or magetuximab. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with the following: trastuzumab or pertuzumab or a combination thereof. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with magetuximab. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with the following: at least one trastuzumab biosimilar or at least one pertuzumab biosimilar or a combination thereof or at least one magetuximab biosimilar. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with at least one trastuzumab biosimilar or at least one pertuzumab biosimilar or a combination thereof. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 in combination with at least one magetuximab biosimilar.

[0139] In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with: at least one HER2 antibody, at least one HER2 dimerization inhibitor antibody, or a combination thereof, or magetuximab. In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with: at least one trastuzumab or at least one pertuzumab or a combination thereof, or at least one magetuximab. In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with: at least one trastuzumab biosimilar or at least one pertuzumab biosimilar or a combination thereof, or at least one magetuximab biosimilar.

[0140] In some embodiments, the HER2 targeted therapy is a HER2 targeted antibody-drug conjugate that specifically binds to HER2. In some embodiments, the HER2 targeted antibody-drug conjugate is, for example, trastuzumab emtansine (T-DM1). or trastuzumab delutec (trastuzumab-delutec) or its biosimilars.

[0141] In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure in combination with at least one HER2 targeting antibody-drug conjugate. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure in combination with trastuzumab emtansine (T-DM1) or trastuzumab delutec (trastuzumab-delutec) In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein and trastuzumab emtansine (T-DM1). Biosimilars or trastuzumab deludec (trastuzumab-deludec) Combinations of biosimilars. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein and trastuzumab emtansine (T-DM1). Or a combination of biosimilars thereof. In some embodiments, the combination comprises at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein and trastuzumab (trastuzumab-delutec) or a combination of biosimilars thereof.

[0142] 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 trastuzumab emtansine (T-DM1). or trastuzumab delutec (trastuzumab-delutec) Combined application.

[0143] In some embodiments, the HER2 targeted therapy is a HER2 small molecule inhibitor. In some embodiments, the HER2 small molecule inhibitor is, for example, tucatinib, neratinib, or lapatinib. In some embodiments, the combination includes a combination of at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein and at least one HER2 small molecule inhibitor. In some embodiments, the combination includes a combination of at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein and tucatinib, neratinib, or lapatinib.

[0144] 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 small molecule inhibitor (e.g., such as tucatinib, neratinib, or lapatinib).

[0145] In some embodiments, the STING agonist antibody-drug conjugate targeting HER2 enhances the efficacy of HER2 targeted therapy.

[0146] Immunotherapy

[0147] Any suitable immunotherapy is contemplated herein for the combination and method disclosed herein. In some embodiments, immunotherapy is an immune checkpoint inhibitor, including but not limited to immune checkpoint molecule binding protein, small molecule inhibitor, antibody, antibody derivative (including Fab fragment and scFv), antibody-drug conjugate, antisense oligonucleotide, siRNA, nucleic acid aptamer, peptide and peptide mimetic. Inhibitory nucleic acids that reduce the expression and / or activity of immune checkpoint molecules can also be used in the combination and method disclosed herein.

[0148] In some embodiments, immune checkpoint inhibitors reduce the expression or activity of one or more immune checkpoint proteins. In other embodiments, immune checkpoint inhibitors reduce the interaction between one or more immune checkpoint proteins and their ligands. See, for example, US20160101 128.

[0149] In some embodiments, immune checkpoint inhibitors suitable for use in the combinations and methods of the present disclosure are monoclonal antibodies, humanized antibodies, fully human antibodies, fusion proteins, or combinations thereof.

[0150] In some embodiments, the composition includes at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein in combination with at least one immune checkpoint inhibitor. In some embodiments, the composition includes at least one STING agonist antibody-drug conjugate targeting HER2 disclosed herein in combination with the following: a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof.

[0151] In some embodiments, the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

[0152] In some embodiments, the PD-1 inhibitor or PD-L1 inhibitor is pembrolizumab (MK-3475), nivolumab (BMS-936558), pidilizumab (CT-011), AMP-224, MDX-1105, durvalumab (MEDU4736), MPDL3280A, BMS-936559, IPH2101, TSR-042, TSR-022, cemiplizumab, ipilimumab, lirilumab, atezolizumab, avelumab, dotalimumab, tesimumab, or a combination thereof.

[0153] In some embodiments, the immune checkpoint inhibitor is avelumab, durvalumab, dotarizumab, pembrolizumab, cemiplizumab, nivolumab, or atezolizumab.

[0154] In some embodiments, the immune checkpoint inhibitor is pembrolizumab, dotalizumab, nivolumab, or atezolizumab. In some embodiments, the immune checkpoint inhibitor is pembrolizumab. In some embodiments, the immune checkpoint inhibitor is dotalizumab.

[0155] In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure and at least one immune checkpoint inhibitor. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure and at least one PD-1 inhibitor or at least one PD-L1 inhibitor.

[0156] In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure and avelumab, durvalumab, dotalimumab, pembrolizumab, cemiplimab, nivolumab or atezolizumab. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure and dotalimumab. In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure and pembrolizumab.

[0157] In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with at least one immune checkpoint inhibitor. In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with at least one PD-1 inhibitor or at least one PD-L1 inhibitor.

[0158] In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with avelumab, durvalumab, dotalimumab, pembrolizumab, cemiplimab, nivolumab, or atezolizumab. In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with dotalimumab. In some embodiments, a combination comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with pembrolizumab.

[0159] In some embodiments, STING agonist antibody-drug conjugates targeting HER2 enhance the efficacy of immunotherapy (e.g., immune checkpoint inhibitors).

[0160] How to use

[0161] In some embodiments, the present disclosure provides a method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0162] In some embodiments, the present disclosure provides a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0163] In some embodiments, the present disclosure provides a method of treating or preventing a disease or condition in a subject in need thereof, comprising administering to the subject at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0164] In some embodiments, the present disclosure provides a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0165] In some embodiments, the present disclosure provides a method of activating or enhancing STING activity in a subject, comprising administering to the subject a combination therapy disclosed herein, i.e., at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0166] In some embodiments, 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.

[0167] In some embodiments, 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.

[0168] In some embodiments, the present disclosure provides a combination therapy disclosed herein for use in treating or preventing a disease or disorder in a subject in need thereof.

[0169] In some embodiments, the present disclosure provides a combination therapy disclosed herein for use in treating a disease or condition in a subject in need thereof.

[0170] In some embodiments, the present disclosure provides a combination therapy disclosed herein for use in treating a STING-mediated disease or disorder in a subject.

[0171] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for treating or preventing a disease or disorder in a subject in need thereof.

[0172] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for treating a disease or condition in a subject in need thereof.

[0173] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for treating cancer in a subject in need thereof.

[0174] In some embodiments, 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.

[0175] In some embodiments, the present disclosure provides for the use of a combination therapy disclosed herein in the manufacture of a medicament for treating a disease or condition in a subject in need thereof.

[0176] In some embodiments, 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.

[0177] In some embodiments, the present disclosure provides for use of a combination therapy disclosed herein in the manufacture of a medicament for treating a STING-mediated disease or disorder in a subject.

[0178] In some embodiments, the present disclosure provides for use of a combination therapy disclosed herein in the manufacture of a medicament for treating cancer in a subject in need thereof.

[0179] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for the treatment or prevention of a disease or disorder in a subject in need thereof.

[0180] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for the treatment of a disease or disorder in a subject in need thereof.

[0181] In some embodiments, the present disclosure provides for the use of a combination therapy disclosed herein for treating a STING-mediated disease or disorder in a subject.

[0182] In some embodiments, the present disclosure provides for the use of the combination therapies disclosed herein for the treatment of cancer in a subject in need thereof.

[0183] In some embodiments, the combination includes at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor.

[0184] In some embodiments, a combination therapy disclosed herein is administered to a subject.

[0185] In some embodiments, the present disclosure provides a method for treating or preventing a disease or pathology in a subject in need thereof, comprising administering to the subject an effective amount of at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor; wherein the STING agonist antibody-drug conjugate targeting HER2 releases one or more therapeutic agents upon biodegradation.

[0186] In some embodiments, the present disclosure provides a method of treating a disease or condition 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-targeting STING agonist antibody-drug conjugate releases one or more therapeutic agents upon biodegradation.

[0187] In some embodiments, the disease or condition is cancer.

[0188] In some embodiments, the cancer is selected from 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 (CRC), 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, stomach cancer, and gastroesophageal junction cancer.

[0189] In some embodiments, the cancer is selected from breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0190] In some embodiments, the combination therapies disclosed herein can be used 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 embodiments, the combination therapy disclosed herein can be used to treat, prevent, delay the progression of breast cancer or otherwise improve its symptoms. In some embodiments, breast cancer is metastatic breast cancer. In some embodiments, breast cancer is HER2 positive (HER2+) breast cancer. In some embodiments, breast cancer is HER2 negative (HER2-) breast cancer. In some embodiments, HER2+ breast cancer is metastatic HER2+ breast cancer. In some embodiments, HER2- breast cancer is metastatic HER2- breast cancer.

[0192] In some embodiments, the subject with breast cancer has received at least one, at least two, at least three or at least four prior breast cancer treatments. In some aspects, the subject has received three or more breast cancer treatments. In some aspects, the subject with HER2+ metastatic breast cancer has received three or more breast cancer treatments.

[0193] In some embodiments, the combination therapies disclosed herein can be used to treat, prevent, delay progression of, or otherwise ameliorate symptoms of gastric cancer.

[0194] In some embodiments, the combination therapies disclosed herein may be used to treat, prevent, slow progression of, or otherwise ameliorate symptoms of gastroesophageal junction cancer.

[0195] In some embodiments, the combination therapies disclosed herein may be used to treat, prevent, delay progression of, or otherwise ameliorate symptoms of colorectal cancer.

[0196] In some embodiments, the combination therapies disclosed herein can be used to treat, prevent, delay progression of, or otherwise ameliorate symptoms of non-small cell lung cancer (NSCLC).

[0197] In some embodiments, the subject has a recurrent or metastatic solid tumor and has HER 2+ expression.

[0198] In some embodiments, the disease or condition is a precancerous syndrome.

[0199] In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor can be used 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.

[0200] In some embodiments, combination therapies including at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy can be used 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.

[0201] In some embodiments, combination therapies comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one immune checkpoint inhibitor can be used 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.

[0202] In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 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 progression of, or otherwise improve symptoms of: breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0203] In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 is administered in combination with trastuzumab or pertuzumab or a combination thereof or magetuximab 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.

[0204] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one trastuzumab biosimilar or at least one pertuzumab biosimilar or a combination thereof or magetuximab for the treatment, prevention, delay of progression, or otherwise amelioration of symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0205] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure and at least one HER2-targeting antibody-drug conjugate (trastuzumab emtansine (T-DM1) or trastuzumab delutec (trastuzumab-delutec) ) are administered in combination to treat, prevent, delay the progression of, or otherwise ameliorate the symptoms of: breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0206] In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with trastuzumab or a trastuzumab biosimilar for the treatment, prevention, delay of progression, or otherwise improvement of 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 STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with trastuzumab or a trastuzumab biosimilar for the treatment, prevention, delay of progression, or otherwise improvement of symptoms of: breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer. or trastuzumab emtansine (T-DM1) In some embodiments, the combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with trastuzumab (trastuzumab-delutocan) 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. or trastuzumab delutec (trastuzumab-delutec) The invention relates to a method 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.

[0207] In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with trastuzumab or a trastuzumab biosimilar for the treatment, prevention, delay of progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise amelioration of its symptoms. In some embodiments, a combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 of the present disclosure is administered in combination with trastuzumab or a trastuzumab biosimilar for the treatment, prevention, delay of progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise amelioration of its symptoms in subjects who have received at least one, at least two, at least three, or at least four prior breast cancer treatments.

[0208] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure and trastuzumab emtansine (T-DM1) is used. or trastuzumab emtansine (T-DM1) Biosimilars are administered in combination to treat, prevent, delay progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise improve its symptoms. In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with trastuzumab emtansine (T-DM 1). or trastuzumab emtansine (T-DM 1) The biosimilars are administered in combination to treat, prevent, delay progression of, or otherwise ameliorate symptoms of HER2-positive (HER2+) metastatic breast cancer in subjects who have received at least one, at least two, at least three, or at least four prior breast cancer treatments.

[0209] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is combined with trastuzumab (trastuzumab-delutec). or trastuzumab delutec (trastuzumab-delutec) Biosimilars are administered in combination 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 HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with trastuzumab (trastuzumab-delutec). or trastuzumab delutec (trastuzumab-delutec) The biosimilars are administered in combination to treat, prevent, delay progression of, or otherwise ameliorate symptoms of HER2-positive (HER2+) metastatic breast cancer in subjects who have received at least one, at least two, at least three, or at least four prior breast cancer treatments.

[0210] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with at least one HER2 small molecule inhibitor (e.g., tucatinib, neratinib, or lapatinib) for the treatment, prevention, delay of progression, or otherwise amelioration of symptoms of breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0211] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure in combination with at least one immune checkpoint inhibitor can be used to treat, prevent, delay progression of, or otherwise improve symptoms of: breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0212] In some embodiments, the composition therapy includes 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, which can be used to treat, prevent, delay the progression of, or otherwise improve the symptoms of: breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer.

[0213] In some embodiments, a combination therapy 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 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.

[0214] In some embodiments, a combination therapy 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 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.

[0215] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with avelumab, durvalumab, dotalimumab, pembrolizumab, cemiplizumab, nivolumab, or atezolizumab for the treatment, prevention, delay of progression, or otherwise amelioration of 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-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dotalimumab for the treatment, prevention, delay of progression, or otherwise amelioration of 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-targeting 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.

[0216] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dotalitamab for the treatment, prevention, delay of progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise amelioration of its symptoms. In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with dotalitamab for the treatment, prevention, delay of progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise amelioration of its symptoms in subjects who have received at least one, at least two, at least three, or at least four prior breast cancer treatments.

[0217] In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with pembrolizumab for treating, preventing, delaying the progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise improving its symptoms. In some embodiments, a combination therapy comprising at least one HER2-targeting STING agonist antibody-drug conjugate of the present disclosure is administered in combination with pembrolizumab for treating, preventing, delaying the progression of HER2-positive (HER2+) metastatic breast cancer, or otherwise improving its symptoms in subjects who have received at least one, at least two, at least three, or at least four prior breast cancer treatments.

[0218] In some embodiments, the method includes confirming the HER2 score of the subject by IHC test or FISH (fluorescence in situ hybridization negative) measurement before using the combination therapy disclosed herein for treatment, to identify or otherwise refine (e.g., stratify) the patient population suitable for therapeutic administration of the combination therapy disclosed herein. The IHC test measures the amount of HER2 receptor protein on the cell surface in a cancer tissue sample (e.g., a breast cancer tissue sample or a gastric cancer sample), and the detected cell surface HER2 receptor level is designated as a HER2 score of 0, 1+, 2+, or 3+. If the subject's HER2 score is in the range of 0 to 1+, the cancer is considered to be "HER2 negative". If the score is 2+, the cancer is referred to as "marginal", and a score of 3+ indicates that the cancer is "HER2 positive".

[0219] In some embodiments, by immunohistochemistry (IHC) analysis detection performed on the test cell colony, the subject is confirmed as having 2+ or 3+ scores of HER2 expression, and the subject is also ER positive. In some embodiments, by immunohistochemistry (IHC) analysis detection performed on the test cell colony, the subject is confirmed as having 2+ or 3+ scores of HER2 expression, and the subject is also ER negative. In some embodiments, the subject is confirmed as having 3+ scores of HER2 expression or having evidence of gene amplification detected by FISH. In some embodiments, the subject is confirmed as having 2+ scores of HER2 expression or having evidence of gene amplification detected by FISH. In some embodiments, the subject is confirmed as having 1+ scores of HER2 expression or having evidence of gene amplification detected by FISH. In some embodiments, the subject is confirmed as having high HER2 expression. In some embodiments, the subject is confirmed as having low HER2 expression. In some embodiments, the test cell colony is derived from fresh, unfrozen tissue of a biopsy sample. In some embodiments, the test cell colony is from the frozen tissue of a biopsy sample.

[0220] In some embodiments, the combination therapy disclosed herein including at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor can be used to treat, prevent, delay the progression of, or otherwise improve symptoms of, breast cancer, gastric cancer, gastroesophageal junction cancer, non-small cell lung cancer (NSCLC), or colorectal cancer 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.

[0221] In some embodiments, the combination therapy disclosed herein comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor can be used 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 patients with HER2 IHC 1+ or HER2 IHC 2+.

[0222] In some embodiments, a combination comprising a STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor disclosed herein can be used 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 patients with HER2 IHC 1+ or HER2 IHC 2+.

[0223] In some embodiments, the combination of a STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor used in any embodiment of the methods and uses provided herein can be administered at any stage of the disease. For example, the combination therapy can be administered to a patient with cancer at any stage from early stage to metastatic.

[0224] The combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immune checkpoint inhibitor used in any embodiment of these methods and uses can be administered without another therapeutic agent or further combined with one or more chemotherapeutic agents or other agents. In some embodiments, the additional agent is any toxin 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 a standard first-line chemotherapy agent, such as, for example, trastuzumab emtansine (Hercel), lapatinib, anastrozole, letrozole, exemestane, everolimus, fulvestrant, tamoxifen, toremifene, megestrol acetate, fluoxymesterone, ethinyl estradiol, paclitaxel, capecitabine, gemcitabine, eribulin, vinorelbine, cyclophosphamide, carboplatin, docetaxel, albumin-bound In some embodiments, the additional agent is Herceptin (enmetuzumab).

[0225] Combination therapies and formulations

[0226] The combination therapies of the present disclosure may be administered together or separately in the form of a single pharmaceutical composition, and when administered separately, they may occur simultaneously or sequentially in any order. The amounts of the components of the combination of the present disclosure and the relative times of administration are selected to achieve the desired combined therapeutic effect.

[0227] The compositions of the present disclosure can be administered once or according to a dosage regimen, wherein multiple dosages are administered at different time intervals within a given time period. For example, the dosage can be administered daily, weekly, biweekly, monthly, every two months, every 6 months or annually. The dosage can be administered until the desired therapeutic effect is reached or administered indefinitely to maintain the desired therapeutic effect. The suitable dosage regimen of the compositions of the present disclosure depends on the pharmacokinetic properties of the compositions, such as absorption, distribution and half-life, which can be determined by those skilled in the art. In addition, the suitable dosage regimen of the compositions of the present disclosure (including the duration of administering such a regimen) depends on the severity of the disease or lesion treated, the disease or lesion treated, the age and physical condition of the patient treated, the medical history of the patient treated, the nature of the simultaneous 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 those skilled in the art that the suitable dosage regimen may need to be adjusted according to the response of individual patients to the dosage regimen or the needs of individual patients over time.

[0228] It will be understood that administration of the conjugates and HER2 targeted therapies or immune checkpoint inhibitors in the disclosed combinations will be administered with suitable carriers, excipients, and other agents incorporated into formulations to provide improved transfer, delivery, tolerability, etc. A variety of suitable formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences (15th edition, Mack Publishing Company, Easton, PA (1975)), particularly Chapter 87 by Blaug and Seymour.

[0229] 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 therapies or one or more immune checkpoint inhibitors disclosed herein.

[0230] In some embodiments, the pharmaceutical composition is in bulk or unit dosage form. The unit dosage form is any of a variety of forms, including, for example, a capsule, an IV bag, a tablet, a single pump on an aerosol inhaler, or a vial. The amount of active ingredient (e.g., a conjugate disclosed herein) in the unit dosage composition is an effective amount and varies according to the specific treatment involved. It will be appreciated by those skilled in the art that it is sometimes necessary to make conventional changes to the dosage depending on the age and condition of the patient. The dosage also depends on the route of administration.

[0231] A pharmaceutical composition is formulated to be compatible with its intended route of administration.Examples of routes of administration include parenteral, such as intravenous, intradermal, subcutaneous, oral (eg, inhalation), transdermal (ie, topical), transmucosal, and rectal administration.

[0232] In some embodiments, the pharmaceutical composition is in bulk or unit dosage form. The unit dosage form is any of a variety of forms, including, for example, a capsule, an IV bag, a tablet, a single pump on an aerosol inhaler, or a vial. The amount of active ingredient (e.g., a conjugate disclosed herein) in the unit dosage composition is an effective amount and varies according to the specific treatment involved. It will be appreciated by those skilled in the art that it is sometimes necessary to make conventional changes to the dosage depending on the age and condition of the patient. The dosage also depends on the route of administration.

[0233] 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 therapies or one or more immune checkpoint inhibitors disclosed herein.

[0234] In some embodiments, a combination of a STING agonist antibody-drug conjugate targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor and an additional agent is formulated into a single therapeutic composition, and the components are administered simultaneously. Alternatively, the STING agonist antibody-drug conjugate targeting HER2, the HER2 targeted therapy or immune checkpoint inhibitor and (if any) additional agents are separated from each other, for example, each formulated into a separate therapeutic composition, and can be administered simultaneously, or at different times during the treatment regimen. For example, a STING agonist antibody-drug targeting HER2 is administered before the administration of HER2 targeted therapy or immune checkpoint inhibitors; a STING agonist antibody-drug targeting HER2 is administered after the administration of HER2 targeted therapy or immune checkpoint inhibitors. As described herein, a combination of a STING agonist antibody-drug targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor is administered in a single dose or multiple doses.

[0235] Dosage and Administration

[0236] The combination therapy provided herein includes a combination of a STING agonist antibody-drug targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor administered in a single dose or multiple doses, which is administered in an amount sufficient to exert a therapeutically useful effect. In some embodiments, the active agent is administered in an amount that does not cause adverse side effects in the patient being treated, or in an amount that minimizes or reduces the observed side effects compared to the dose and amount required for single treatment with one of the above-mentioned agents. For example, a combination therapy is administered in a single dose or multiple doses, and the combination therapy includes a combination of a STING agonist antibody-drug targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor. Therefore, compared with the amount of HER2 targeted therapy or immune checkpoint inhibitor administered alone or using a known method, the amount of HER2 targeted therapy or immune checkpoint inhibitor that can be applied is reduced in the combination therapy provided herein, while achieving substantially the same or improved therapeutic efficacy. Because the dose that can be applied is reduced, side effects associated with the administration of HER2 targeted therapy or immune checkpoint protein antibodies, such as immune-related adverse events described herein or elsewhere, can be reduced, minimized or avoided.

[0237] The precise amount of active agent administered to a subject can be determined within the level of those skilled in the art, including a STING agonist antibody-drug conjugate targeting HER2 and a HER2 targeted therapy or immune checkpoint inhibitor. The dosage of such an agent in a combination therapy can be selected based on the standard dosing regimen of the agent under a given administration route.

[0238] It should be understood that the exact dosage and duration of treatment depend on the tissue or tumor being treated, and can be determined empirically using known test protocols, or inferred by in vivo or in vitro test data, and / or can be determined by known dosing regimens for specific agents. It should be noted that concentrations and dosage values ​​may also vary with the age of the individual being treated, the individual's weight, the route of administration, and / or the extent or severity of the disease, as well as other factors that can be considered within the level of skilled medical practitioners. Generally, the dosage regimen is selected to limit toxicity. It should be noted that the treating physician will know how and when to terminate, interrupt or adjust the therapy to a lower dose to cope with toxicity or bone marrow, liver or kidney or other tissue dysfunction. Conversely, if the clinical response is not sufficient (excluding toxic side effects), the treating physician will also know how and when to adjust the treatment to a higher level. It should also be further understood that for any particular subject, the specific dosage regimen should be adjusted over time according to personal needs and the professional judgment of the person administering or supervising the administration of the formulation, and the concentration ranges described herein are exemplary only and are not intended to limit their scope.

[0239] In some embodiments, the HER2-targeting STING agonist antibody-drug conjugate combination is administered in a therapeutically effective amount to reduce tumor volume.

[0240] The amount of the STING agonist antibody-drug conjugate targeting HER2 administered to treat a disease or condition (e.g., cancer or solid tumor) can be determined by standard clinical techniques. In addition, in vitro assays and animal models can be used to help confirm the optimal dosage range. The exact dosage can be determined empirically and may depend on the route of administration, the type of disease being treated, and the severity of the disease.

[0241] HER2 targeted therapy or immune checkpoint inhibitors can be provided in a therapeutically effective amount of a specific dosing regimen. The therapeutically effective concentration can be determined empirically by testing the compound in known in vitro and in vivo systems (e.g., assays provided herein). The concentration of the selected HER2 targeted therapy and immune checkpoint inhibitor in the composition depends on the absorption, inactivation and excretion rate of the complex, the physicochemical properties of the complex, the dosing regimen and the amount of administration, and other factors known to those skilled in the art.

[0242] The amount of the selected HER2 targeted therapy or immune checkpoint inhibitor applied to treat cancer can be determined by standard clinical techniques or other methods described herein. In addition, in vitro assays and animal models can be used to help confirm the optimal dosage range. Therefore, the exact dosage can be determined empirically and may depend on the route of administration, the type of cancer treated, and the progression of the disease. If necessary, the specific dosage and duration and treatment regimen can be determined or inferred empirically. The dosage level can be determined based on a variety of factors, such as individual weight, general health, age, activity of the specific compound used, sex, diet, administration time, excretion rate, drug combination, severity and course of the disease, and the patient's mood and judgment of the treating physician.

[0243] The combination of the present disclosure can also be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and inhalation administration. Parenteral administration refers to routes of administration other than enteral, transdermal, or inhalation, and is, for example, by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to inhalation through the mouth or through the nasal cavity to be applied to the patient's lungs. Topical administration includes application to the skin.

[0244] In some embodiments, the HER2-targeting STING agonist antibody-drug conjugate and the HER2-targeted therapy or immune checkpoint inhibitor are administered by infusion every week, every two weeks, every three weeks, every four weeks, every five weeks, every six weeks, every seven weeks, or every eight weeks.

[0245] In some embodiments, the HER2-targeting STING agonist antibody-drug conjugate and the HER2-targeted therapy or immune checkpoint inhibitor are administered by infusion every three weeks or every four weeks.

[0246] In some embodiments, the STING agonist antibody-drug targeting HER2 and the HER2 targeted therapy or immune checkpoint inhibitor are administered by infusion at the same time. In some embodiments, the STING agonist antibody-drug targeting HER2 is administered by infusion before the administration of the HER2 targeted therapy or immune checkpoint inhibitor. In some embodiments, the STING agonist antibody-drug targeting HER2 is administered by infusion after the administration of the HER2 targeted therapy or immune checkpoint inhibitor. As described herein, the combination of the STING agonist antibody-drug targeting HER2 and the HER2 targeted therapy or immune checkpoint inhibitor can be administered in a single dose or multiple doses.

[0247] The frequency and time of administration and the amount of dosage can be regularly applied within the administration cycle to maintain the continuous and / or long-term effect of the active agent within the required time length. The combination provided can be applied hourly, daily, weekly, monthly, annually or once. The time length of the administration cycle can be determined empirically and depends on the disease being treated, the severity of the disease, the specific patient and other considerations within the treating physician's skill level. The time length for the combination therapy treatment provided herein can be one week, two weeks, one month, several months, one year, several years or longer.

[0248] In some embodiments, exemplary dosages of intravenously administered immune checkpoint inhibitors (e.g., anti-immune checkpoint protein antibodies) can be used as a starting point for determining an appropriate dosage. The dosage level can be determined based on a variety of factors, such as individual weight, general health, age, activity of the specific compound used, sex, diet, administration time, excretion rate, drug combination, severity and course of the disease, the patient's mood caused by the disease, and the judgment of the treating physician.

[0249] It will be appreciated that the amount administered will depend on the type of cancer being treated, the route of administration, and the tolerance of possible side effects. If necessary, the dosage can be determined empirically.

[0250] For intravenous administration, one or more or all agents used in the combination therapy can be administered by push or injection, by infusion or via a combination thereof. The infusion time can be about 1 minute to 3 hours, for example, about 1 minute to about 2 hours, about 1 minute to about 60 minutes, about 1 minute to about 90 minutes, or about 1 minute to about 120 minutes. The agent can be administered by simultaneous infusion or by subsequent infusion. For example, the administered agents are administered separately and provided in separate bags for separate infusion. In some embodiments, HER2 targeting antibody-drug conjugate compositions and HER-2 targeted therapies or immune checkpoint inhibitor compositions are separately formulated and administered.

[0251] The STING agonist antibody-drug conjugate targeting HER2 can be administered before, simultaneously or nearly simultaneously, sequentially or intermittently with HER2 targeted therapy or immune checkpoint inhibitors. For example, HER2 targeted antibody-drug conjugates and HER2 targeted therapy or immune checkpoint inhibitors (e.g., anti-immune checkpoint protein antibodies (e.g., anti-CTLA4 or anti-PD-1 antibodies)) can be administered together or separately.

[0252] In some embodiments, the STING agonist antibody-drug conjugate targeting HER2 is administered before HER2 targeted therapy or immune checkpoint inhibitors. For example, the STING agonist antibody-drug conjugate targeting HER2 is administered up to about 48 hours before the administration of HER2 targeted therapy or immune checkpoint inhibitors. For example, the STING agonist antibody-drug conjugate targeting HER2 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 administration of HER2 targeted therapy or immune checkpoint inhibitors.

[0253] In some embodiments, the STING agonist antibody-drug conjugate targeting HER2 is administered after HER2 targeted therapy or immune checkpoint inhibitors. For example, the STING agonist antibody-drug conjugate targeting HER2 is administered up to about 48 hours after the administration of HER2 targeted therapy or immune checkpoint inhibitors. For example, the STING agonist antibody-drug conjugate targeting HER2 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 after the administration of HER2 targeted therapy or immune checkpoint inhibitors.

[0254] The frequency and time of administration and the amount of the dosage can be regularly administered within the administration cycle to maintain the sustained and / or long-term effect of the active agent for the desired length of time, and it is not necessary to be the same for the STING agonist antibody-drug conjugate targeting HER2 and the HER2 targeted therapy or immune checkpoint inhibitor. The composition of each active agent or its combination provided can be administered hourly, daily, weekly, monthly, annually or once. The length of the administration cycle can be determined empirically and depends on the disease being treated, the severity of the disease, the patient's mood, and other considerations within the skill level of the treating physician. The length of time for treatment by the combination therapy provided herein can be one week, two weeks, one month, several months, one year, several years or longer.

[0255] For example, the STING agonist antibody-drug conjugate targeting HER2 is administered 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. During the course of treatment, the dosage can be divided into multiple administration cycles. For example, the STING agonist antibody-drug conjugate targeting HER2 can be administered at the frequency for a period of about one month, two months, three months, four months, five months, six months, one year, or longer. The frequency of administration can be the same throughout the cycle or can be different. For example, an exemplary dosage frequency is twice a week at least in the first week of the administration cycle. After the first week, the frequency can continue to be twice a week, can be increased to more than twice a week, or can be reduced to no more than once a week. The specific dosage frequency and administration cycle can be determined by those skilled in the art based on the specific dose administered, the disease or condition being treated, the severity of the disease or condition, the age of the subject, and other similar factors.

[0256] HER2 targeted therapy or immune checkpoint inhibitors can be administered at the same frequency or at different frequencies. For example, before each administration of HER2 targeted therapy or immune checkpoint inhibitor, a STING agonist antibody-drug conjugate targeting HER2 is administered within no more than 48 hours. For example, a dose of HER2 targeted therapy or immune checkpoint inhibitor is administered 24 to 48 hours after each dose of STING agonist antibody-drug conjugate targeting HER2 is administered. In certain embodiments, the frequency of administration of HER2 targeted therapy or immune checkpoint inhibitors is lower than that of STING agonist antibody-drug conjugate targeting HER2, but a dose of HER2 targeted antibody-drug conjugate is administered before each dose of HER2 targeted therapy or immune checkpoint inhibitor. For example, HER2 targeted therapy or immune checkpoint inhibitors are administered twice a week, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months, and are administered in a manner that a STING agonist antibody-drug conjugate targeting HER2 is administered before this. In another embodiment, a dose of HER2 targeted therapy or immune checkpoint inhibitor is administered before each dose of the STING agonist antibody-drug conjugate targeting HER2. In certain embodiments, the frequency of administration of HER2 targeted therapy or immune checkpoint inhibitor is higher than that of the STING agonist antibody-drug conjugate targeting HER2. For example, HER2 targeted therapy or immune checkpoint inhibitor is administered twice a week, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, once every 2 months, once every 3 months, once every 4 months, once every 5 months, or once every 6 months, and is administered in a manner that the STING agonist antibody-drug conjugate targeting HER2 is administered after some but not all HER2 targeted therapy doses or checkpoint inhibitor doses.

[0257] If disease symptoms persist despite cessation of treatment, treatment may be continued for an additional period of time.During treatment, one may monitor for evidence of disease and / or treatment-related toxicity or side effects.

[0258] The administration cycle of the STING agonist antibody-drug conjugate targeting HER2 and / or HER2 targeted therapy and / or immune checkpoint inhibitor can be customized to add a period of time for stopping treatment to provide a rest period away from drug exposure. The length of time for stopping treatment can be a predetermined time, or can be determined empirically depending on how the patient responds or on observed side effects. For example, treatment can be stopped for a week, two weeks, a month, or several months. Typically, the period of time for stopping treatment is included in the patient's dosing regimen cycle.

[0259] An exemplary dosing regimen is a 21 or 28 day treatment cycle or administration cycle. An agent such as a STING agonist antibody-drug conjugate targeting HER2 disclosed herein can be administered on day 1, followed by administration of a HER2 targeted therapy or immune checkpoint inhibitor disclosed herein, such as a HER2 targeted therapy or immune checkpoint protein antibody, on day 2, followed by no administration for 19 or 26 days. It should be understood that the above description is for exemplary purposes only, and that variants described above may also be employed. Further, a similar administration cycle may be applied to all administered agents, or each administered agent may be employed in its own dosing regimen in the combination therapy provided herein.

[0260] Determining the precise administration cycle and dosing schedule is within the technical level of those skilled in the art. As mentioned above, the administration cycle can be any desired length of time. Therefore, an administration cycle of 21 days or 28 days can be repeated for any length of time. The administration cycle and dosing regimen that meet the patient's needs can be adopted depending on the individual considerations of the specific patient and the disease being treated within the technical level of the treating physician.

[0261] Diagnostic and preventive preparations

[0262] The conjugates disclosed herein and HER2 targeted therapies or immune checkpoint inhibitors are used for diagnostic and preventive preparations. In one embodiment, the STING agonist antibody-drug conjugates and HER2 targeted therapies or immune checkpoint inhibitors targeting HER2 disclosed herein are administered to patients with a risk of developing one or more of the above diseases (such as but not limited to cancer). Genotype, serology or biochemical markers can be used to determine the susceptibility of a patient or organ to one or more of the above indications.

[0263] In some embodiments, the STING agonist antibody-drug conjugates and immune checkpoint inhibitors targeting HER2 disclosed herein are administered to human individuals diagnosed with clinical indications associated with one or more of the above diseases (e.g., but not limited to cancer). After diagnosis, the STING agonist antibody-drug conjugates and HER2 targeted therapies or immune checkpoint inhibitors targeting HER2 disclosed herein are administered to reduce or reverse the effects of clinical indications associated with one or more of the above diseases.

[0264] 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 therapy or an immune checkpoint inhibitor comprises measuring the state of certain features in a tumor sample obtained from the patient, and identifying a patient suitable for treatment based on the state of the certain features in the tumor sample.

[0265] Antibodies disclosed herein can also be used to detect HER2 in patient samples, and therefore can be used as diagnostic agents. For example, HER2 antibodies disclosed herein are used for in vitro assays, such as ELISA, to detect HER2 levels in patient samples.

[0266] In some embodiments, HER2 antibody disclosed herein is fixed on a solid support (e.g., a hole in a microtiter plate). Immobilized antibody is used as a capture antibody for any HER2 that may be present in a test sample. Before the immobilized antibody is contacted with a patient sample, the solid support is rinsed and treated with a blocking agent (e.g., milk protein or albumin) to prevent nonspecific adsorption of the analyte.

[0267] The wells are then treated with a test sample suspected of containing antigen or with a solution containing a standard amount of antigen. Such samples are, for example, serum samples from subjects who are suspected of having a pathologically diagnosable circulating antigen level. After rinsing off the test sample or standard sample, the solid support is treated with a second antibody that is detectably labeled. The second antibody of the label is used as a detection antibody. The level of the detectable label is measured and compared with the standard curve developed from the standard sample to determine the concentration of the HER2 antigen in the test sample.

[0268] It should be understood that, based on the result obtained in the in vitro diagnostic assay using HER2 antibody disclosed herein, it is possible to stage the disease of the experimenter based on the expression level of HER2 antigen.For a given disease, blood samples are collected from the experimenter diagnosed as being in each stage of disease progression and / or in each node of disease treatment.Use provides a sample population with statistically significant results for each stage of progress or treatment, to specify the antigen concentration range that may be considered as each stage feature.

[0269] All publications and patent documents cited herein are incorporated herein by reference, just as if each such publication or document was specifically and individually indicated to be incorporated herein by reference. The citation of publications and patent documents is not intended to admit any relevant prior art, nor does it constitute any admission of their contents or dates. The present invention has now been described by way of written description, and those skilled in the art will recognize that the present invention can be practiced in a variety of embodiments, and the foregoing description and the following examples are for illustrative purposes only and are not intended to limit the subsequent claims. Example

[0270] The following examples illustrate the present disclosure. These examples are not intended to limit the scope of the present disclosure, but provide guidance to the skilled person to prepare and use the compounds, compositions and methods of the present disclosure. Although specific embodiments of the present disclosure have been described, it will be appreciated by the skilled person that various changes and modifications may be made without departing from the spirit and scope of the present disclosure.

[0271] It will be appreciated that certain compounds of the present disclosure may be potent immunomodulators and therefore caution should be exercised in their handling.

[0272] Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification.

[0273] abbreviation

[0274] The following abbreviations are used in the following reaction schemes and synthetic examples. This list is not meant to be an all-inclusive list of abbreviations used in this application, as additional standard abbreviations that are readily understood by those skilled in the art of organic synthesis may also be used in the synthetic schemes and examples.

[0275] CHT Ceramic Hydroxyapatite

[0276] CR Fully Response

[0277] HIC Hydrophobic Interaction Chromatography

[0278] HPLC High Performance Liquid Chromatography

[0279] PR Partial Response

[0280] RP-HPLC Reversed Phase High Performance Liquid Chromatography

[0281] SEC Size Exclusion Chromatography

[0282] TCEP (Tris(2-carboxyethyl)phosphine)

[0283] TGD Tumor Growth Delay

[0284] TGI Tumor Growth Inhibition

[0285] TFS Tumor-free survivor

[0286] General information

[0287] Unless otherwise stated, all reagents were purchased from relevant suppliers.

[0288] XMT-1519 (anti-Her2 antibody) is disclosed in US 9,555,112 published on January 31, 2017 and US 9,738,720 published on August 22, 2017, the entire contents of which are incorporated herein by reference.

[0289] The antibody-drug conjugate Enhertu is also known as trastuzumab-delutecan or T-DXd.

[0290] 7.16.4_msIgG2a mab is a mouse IgG2a antibody that recognizes the p185 molecule encoded by the rat neu oncogene (rat HER2 (neu)). Since the humanized antibody XMT-1519 does not bind to rat HER2, conjugates 1 and 2 cannot be used in the rat HER2 EMT6-RHER2-MSA breast cancer tumor model. Therefore, alternative conjugates (7.16.4_msIgG2a conjugate and non-binding control palivizumab_msIgG2a conjugate, respectively conjugate 4 and conjugate 3) are used in the rat HER2 EMT6-RHER2-MSA breast cancer tumor model because they specifically recognize rat HER2 (neu).

[0291] In Phenomenex Gemini 5μm C18 HPLC purification was performed on a 250x10 mm semi-preparative column.

[0292] When applicable, the drug content of the conjugates was determined spectrophotometrically, otherwise RP-HPLC or LC / MS was performed to quantify the drug content.

[0293] The protein content of the antibody-drug conjugates was determined spectrophotometrically or by ELISA.

[0294] The antibody-drug conjugate, drug-loaded scaffold or antibody scaffold is purified (i.e., to remove residual unreacted drug, unconjugated antibody, enzyme or starting material) by ultrafiltration, extensive diafiltration, CHT chromatography or HIC as needed. If necessary, additional purification is performed by SEC or HIC to remove aggregated antibody-drug conjugates. Generally, the purified antibody-drug conjugate contains <5% (w / w) (e.g., <2% (w / w)) of aggregated antibody-drug conjugates, as determined by SEC; <0.5% (w / w) (e.g., <0.1% (w / w)) of free (unconjugated) drug, as determined by RP-HPLC and / or LC-MS / MS; <1% (w / w) of free drug conjugate, as determined by SEC and / or RP-HPLC; and <10% (w / w) (e.g., <1% (w / w)) of unconjugated antibody or antibody fragment, as determined by HIC-HPLC and / or RP-HPLC. Reduced or partially reduced antibodies were prepared using methods 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.

[0295] The drug to antibody ratio (DAR) was determined by measuring the absorbance of the conjugate. DAR values ​​were calculated using the appropriate molar extinction coefficients for the antibody and STING agonist payload.

[0296] Therapeutic agents (including STING agonist ADCs targeting HER2), HER2 targeted therapies, and / or immunotherapies may be administered or dosed at frequencies and intervals disclosed herein. In some aspects, the therapeutic administration regimen may be as follows.

[0297] Administration regimen of therapeutic agents

[0298] QD Single Dose

[0299] QD X 1 Single administration on day 1

[0300] QWK is administered once a week

[0301] QWK X 2 Administer once a week for 2 weeks

[0302] QWK X 3 Administer once a week for 3 weeks

[0303] BIW is applied twice a week

[0304] BIW X 2 Apply twice a week for 2 weeks

[0305] The dose of antibody / payload represents milligrams of antibody / kg body weight and milligrams of payload / kg body weight. For example, a conjugate dose of 0.30 / 0.011 mg / kg represents 0.30 mg of antibody administered per kg body weight and 0.011 mg of conjugate payload administered per kg body weight.

[0306] Tumors were measured twice weekly using digital calipers and tumor volumes were calculated using the following formula: Tumor volume (mm 3 )=(width 2 x length) / 2. Body weights were recorded daily for the first week and twice weekly thereafter. Animals were studied until a single tumor reached a volume of ≥1000 mm 3 , ≥1500mm 3 Or as described. The following formula was used to calculate the percentage of body weight change: Body weight change (%) = ((body weight 研究第x天 -weight 研究第1天 ) / weight 研究第1天)*100. Tumor volumes are reported as mean ± standard error of the mean (SEM). Tumor growth inhibition (%TGI) is defined as the percentage difference in mean tumor volume (MTV) between the treatment group and the control group. Tumor size was measured throughout each efficacy study period to determine tumor growth inhibition (TGI). The percentage of tumor regression was calculated using the following formula: % regression = (1-(mean tumor volume 最终 ) / (mean tumor volume 第1天 ))*100. Partial response (PR) was defined as three consecutive measurements of tumor volume of 50% or less of the volume on day 1, and at least one of the three measurements was equal to or greater than 13.5 mm 3 Complete response (CR) was defined as three consecutive measurements of tumor volume less than 13.5 mm 3 Tumor-free survivors (TFS) were classified as having a CR at the end of the study.

[0307] Example 1: Synthesis of XMT-1519 Conjugate 1 (DAR 8.1)

[0308]

[0309] Conjugate 1 was prepared as described in co-pending application US 17 / 221,341 filed on April 4, 2021. Purified Conjugate 1 had a STING agonist to XMT-1519 ratio of 8.1.

[0310] Example 2: Synthesis of Palivizumab Conjugate 2 (DAR6.8)

[0311]

[0312] Conjugate 2 was prepared as described in co-pending application US 17 / 221,341 filed on April 4, 2021. Purified conjugate 2 had a STING agonist to palivizumab ratio of 6.8.

[0313] Example 3: Synthesis of Palivizumab-msIgG2a Conjugate 3 (DAR7.7)

[0314]

[0315] TCEP-HCl (1.76 mg, 6.17 μmol) was added to a solution of palivizumab-msIgG2a (90 mg, 0.617 μmol) in a buffer of 50 mM HEPES, 1 mM EDTA, pH 7, to make a final antibody concentration of 5 mg / mL, and the mixture was shaken at 37 ° C for 4 hours. Scaffold A (prepared as described in co-pending application US 17 / 221,341 filed on April 4, 2021, 17.45 mg, 7.404 μmol, 1800 μL DMA) was added to the reduced antibody. The resulting mixture was shaken at 37 ° C for 120 minutes. The reaction was quenched with cysteine ​​(15 equivalents, 1.121 mg, 9.255 μmol, 1.12 mL of 50 mM HEPES, 1 mM EDTA, pH 7) and rotated at room temperature for 1 hour. The resulting conjugate 3 (80 mg, 90% yield) was purified by ultrafiltration or CHT chromatography. The purified conjugate 3 had a STING agonist to palivizumab_msIgG2a ratio of 7.7.

[0316] Example 4: Synthesis of 7.16.4_msIgG2a Conjugate 4 (DAR 6.9)

[0317]

[0318] Conjugate 4 was prepared and characterized as described in Example 3, but using 7.16.4_msIgG2a instead of palivizumab_msIgG2a. Purified conjugate 4 had a ratio of STING agonist to 7.16.4_msIgG2a of 6.9.

[0319] Example 5: Tumor Growth Response of Combination Administration of HER2 STING Agonist Antibody-Drug Conjugate and Trastuzumab in SKOV3 Ovarian Cancer

[0320] Female CB.17SCID mice were subcutaneously inoculated with SKOV3 human ovarian cancer cells (10x10 6 cells / mouse). SKOV3 cells are a breast cancer cell line with high HER2 expression. 3 Between (average value = 72-81mm 3 / group), the animals were randomized into treatment groups. Dosing was qd x 1 on day 1 or qwk x 3 on days 1, 8 and 15 as follows: vehicle, conjugate 1 (0.30 / 0.013 mg / kg), a combination of conjugate 1 (0.30 / 0.013 mg / kg) and trastuzumab (3 mg / kg), a combination of conjugate 1 (0.30 / 0.013 mg / kg) and trastuzumab (10 mg / kg), and a combination of conjugate 2 (0.30 / 0.011 mg / kg) and trastuzumab (10 mg / kg). In addition, trastuzumab (10 mg / kg) was administered qwk x 3 starting from days 1, 8 and 15. For all doses, the conjugate was administered intravenously (IV); trastuzumab was administered intraperitoneally (IP); all doses of the conjugate are given as antibody / payload; n=10 per group.

[0321] Figure 1 Results of tumor volume in 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 different dose levels and dosing schedules are provided. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd x 1) resulted in 1 PR and 3 CRs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd x 1) and trastuzumab (3 mg, qd x 1) resulted in 7 CRs and 3 TFSs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qd x 1) and trastuzumab (10 mg, qd x 1) resulted in 2 CRs and 1 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 and 9 CRs and 9 TFSs. Treatment with conjugate 1 (0.30 / 0.013 mg / kg, qwk x 3) and trastuzumab (10 mg, qwk x3) resulted in 2 PRs and 7 CRs and 7 TFSs. The results showed that the addition of trastuzumab synergistically enhanced the efficacy of conjugate 1.

[0322] Embodiment 6: Tumor growth response of HER2STING agonist antibody-drug conjugates administered in combination with trastuzumab or pertuzumab in JIMT-1

[0323] Female CB.17SCID mice were subcutaneously inoculated with JIMT-1 tumor cells (10 x 10 6cells / mouse). JIMT-1 cells are a breast cancer cell line with moderate HER2 expression. 3 Between (average 67.8-71.6mm 3 / group), the animals were randomized into treatment groups. On day 1, the following were dosed qd x 1: vehicle, conjugate 1 (3.0 / 0.13 mg / kg), conjugate 2 (3.0 / 0.11 mg / kg) and trastuzumab (10 mg / kg), conjugate 2 (3.0 / 0.11 mg / kg) and pertuzumab (10 mg / kg), conjugate 1 (3.0 / 0.13 mg / kg) and trastuzumab (3 mg / kg), conjugate 1 (3.0 / 0.13 mg / kg) and trastuzumab (10 mg / kg) , the combination of Conjugate 1 (3.0 / 0.13 mg / kg) and Pertuzumab (3 mg / kg), the combination of Conjugate 1 (3.0 / 0.13 mg / kg) and Pertuzumab (10 mg / kg), the combination of Conjugate 2 (3.0 / 0.11 mg / kg), Trastuzumab (3 mg / kg) and Pertuzumab (3 mg / kg), and the combination of Conjugate 1 (3.0 / 0.13 mg / kg), Trastuzumab (3 mg / kg) and Pertuzumab (3 mg / kg). In addition, the combination of Conjugate 1 (0.30 / 0.013 mg / kg), Conjugate 2 (0.30 / 0.011 mg / kg) and Trastuzumab (3 mg / kg), the combination of Conjugate 2 (0.30 / 0.011 mg / kg) and Pertuzumab (3 mg / kg), the combination of Conjugate 1 (0.30 / 0.013 mg / kg) and Trastuzumab (3 mg / kg), and the combination of Conjugate 1 (0.30 / 0.013 mg / kg) and Pertuzumab (3 mg / kg) were dosed qwk x 3 on days 1, 8, and 15. For all doses, the conjugates were administered intravenously; Trastuzumab and Pertuzumab were administered intraperitoneally; all doses of the conjugates are given as antibody / payload; n=10 per group.

[0324] Figure 2 Results of tumor volume in JIMT-1 tumor-bearing mice treated with vehicle, conjugate 1, a combination of conjugate 1 and trastuzumab, a combination of conjugate 1 and pertuzumab, a combination of conjugate 1 and trastuzumab and pertuzumab, a combination of conjugate 2 and trastuzumab, a combination of conjugate 2 and pertuzumab, and a combination of conjugate 2 and trastuzumab and pertuzumab at different dose levels and dosing schedules are provided.

[0325] All treatment groups showed significant tumor growth delay (TGD) compared to vehicle control. However, only the groups treated with the following achieved the maximum possible TGD (78%): conjugate 1 (3 mg / kg) alone or in combination with trastuzumab and / or pertuzumab, and conjugate 1 (0.3 mg / kg) in combination with trastuzumab or pertuzumab, although these groups differed in the number of regression responses and tumor-free survivors (TFS) they produced. Treatment with conjugate 1 (3 mg / kg, qd x 1) and pertuzumab (3 or 10 mg / kg, qd x 1) resulted in 9 TFS and 8 TFS, respectively, with 10 CRs each. Treatment with conjugate 1 (3 mg / kg, qd x 1) and trastuzumab (3 or 10 mg / kg, qd x 1) each resulted in 3 TFS, with 10 CRs and 9 CRs and 1 PR, respectively. All treatment groups except for Conjugate 2, Trastuzumab and Pertuzumab showed significant TGI compared to vehicle control.

[0326] Example 7: Tumor Growth Response in SNU-5 Patients Administering a Combination of a HER2 STING Agonist Antibody-Drug Conjugate with Trastuzumab or Pertuzumab

[0327] Female CB.17SCID mice were subcutaneously inoculated with SNU-5 tumor cells (10 x 10 6 cells / mouse). SNU-5 is a gastric cancer cell line with low HER2 expression. 3 Between (average 79.7-81.3mm 3 / group), the animals were randomly divided into treatment groups. On day 1, the following were dosed qd x 1: vehicle, conjugate 1 (0.20 / 0.007 mg / kg), trastuzumab (2 mg / kg), pertuzumab (2 mg / kg), conjugate 2 (0.20 / 0.007 mg / kg), the combination of conjugate 1 (0.20 / 0.007 mg / kg) and trastuzumab (2 mg / kg), the combination of conjugate 1 (0.20 / 0.007 mg / kg) and pertuzumab (2 mg / kg), the combination of trastuzumab (2 mg / kg) and pertuzumab (2 mg / kg), and the combination of conjugate 1 (0.20 / 0.007 mg / kg), trastuzumab (2 mg / kg) and pertuzumab (2 mg / kg). In addition, 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) and trastuzumab (2 mg / kg), and the combination of conjugate 1 (0.10 / 0.004 mg / kg) and pertuzumab (2 mg / kg), wherein the conjugate was administered qwk x 3 on days 1, 8, and 15, and trastuzumab or pertuzumab was administered qd x 1 on day 1. For all doses, the conjugate was administered intravenously; trastuzumab and pertuzumab were administered intraperitoneally; all doses of the conjugate are given as antibody / payload; n=10 per group).

[0328] Figure 3 Results of tumor volume in 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 different dose levels and dosing schedules are provided.

[0329] Example 8: Tumor growth response of a combination of HER2 STING agonist antibody-drug conjugate and Enhertu in JIMT-1

[0330] Female CB.17SCID mice were subcutaneously inoculated with JIMT-1 tumor cells (10 x 10 6 cells / mouse). When the tumor volume is 60-100mm 3 Between (average 76.8mm 3 / group), and the animals were randomly divided into treatment groups. Dosing was as follows: vehicle, Conjugate 2 (1.0 / 0.037 mg / kg), Conjugate 1 (1.0 / 0.043 mg / kg), Enhertu (1.0 / 0.026 mg / kg, 3.0 / 0.078 mg / kg or 10.0 / 0.261 mg / kg), the combination of Conjugate 1 (1.0 / 0.043 mg / kg) and Enhertu (1.0 / 0.026 mg / kg), the combination of Conjugate 1 (1.00 / 0.043 mg / kg) and Enhertu (3.0 / 0.078 mg / kg), and the combination of Conjugate 1 (1.00 / 0.043 mg / kg) and Enhertu (10.0 / 0.26 mg / kg), wherein the conjugate was administered intravenously qd x 1 on day 1 and Enhertu was administered intravenously qwk x 2 on days 1 and 8. In addition, on days 1 and 8, the following were administered intravenously: Conjugate 1 (0.30 / 0.013 mg / kg), the combination of Conjugate 1 (0.3 / 0.13 mg / kg) and Enhertu (1.0 / 0.026 mg / kg), the combination of Conjugate 1 (0.3 / 0.13 mg / kg) and Enhertu (3.0 / 0.078 mg / kg), and the combination of Conjugate 1 (0.3 / 0.13 mg / kg) and Enhertu (10.0 / 0.261 mg / kg); all doses of conjugates are given as antibody / payload; n=10 per group.

[0331] Figure 4 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 different dose levels and dosing schedules are provided.

[0332] Example 9: Tumor growth response of HER2 STING agonist antibody-drug conjugates administered in combination with anti-PD-1 in rat HER2EMT6-RHER2-MSA breast cancer

[0333] PD-L1 was upregulated in SKOV3 tumors after treatment with Conjugate 1 ( Figure 7 ), suggesting that combining conjugate 1 with immunotherapy (e.g., PD-1 checkpoint inhibitors) would be effective. After treatment with conjugate 1, upregulation of PD-L1 was observed in both mouse and human cells.

[0334] Female BALB / c mice were subcutaneously inoculated with EMT6-RHER2_MSA (an engineered EMT6 cell line overexpressing rat epidermal growth factor receptor 2 (RHER2)) (5 ​​x 10 6cells / mouse). When the tumor volume is 60-100mm 3 Between (average = 95-109mm 3 / group), the animals were randomly divided into treatment groups. Vehicle, conjugate 3 (0.30 / 0.013mg / kg, 1.0 / 0.04mg / kg or 3.0 / 0.12mg / kg), conjugate 4 (0.30 / 0.012mg / kg, 1.0 / 0.035mg / kg or 3.0 / 0.104mg / kg) were all administered qd x 1 on day 1. Anti-PD-1 RMP1-14 (1mg / kg or 5mg / kg) was administered biw x 2 on days 1, 4, 8 and 11. The combination of conjugate 3 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), the combination of conjugate 3 (1 / 0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), the combination of conjugate 4 (0.30 / 0.012 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), the combination of conjugate 4 (1.0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg / kg), wherein the conjugate is administered qd x 1 on day 1, and anti-PD-1 RMP1-14 is administered biw x 2 on days 1, 4, 8, and 11. For all doses, the conjugate was administered intravenously; anti-PD-1 RMP1-14 was administered intraperitoneally; all doses of the conjugate are given as antibody / payload; n=10 per group.

[0335] Figure 5The results of treating the tumor volume of EMT6-RHER2_MSA tumor-bearing mice with different dose levels and dosing regimens using vehicle, conjugate 3, conjugate 4, anti-PD-1RMP1-14, a combination of conjugate 3 and anti-PD-1RMP1-14, and a combination of conjugate 4 and anti-PD-1RMP1-14 are provided. Treatment with conjugate 3 (1.0 / 0.04mg / kg) resulted in 5 CRs and 5 TFSs. Treatment with conjugate 3 (3.0 / 0.12mg / kg) resulted in 7 CRs and 7 TFSs. Treatment with conjugate 4 (0.3 / 0.012mg / kg) resulted in 5 CRs and 5 TFSs. Treatment with conjugate 4 (1.0 / 0.035mg / kg) resulted in 10 CRs and 10 TFSs. Treatment with conjugate 4 (3.0 / 0.10mg / kg) resulted in 9 CRs and 9 TFSs. 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 TFSs. 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 TFSs. Treatment with conjugate 4 (1.0 / 0.035 mg / kg) and anti-PD-1 RMP1-14 (5 mg) resulted in 10 CRs and 10 TFSs.

[0336] Example 10: Re-challenge study of tumor growth response after combined administration of HER2 STING agonist antibody-drug conjugate and anti-PD-1

[0337] Tumor-free mice from Example 9 (i.e., mice with complete tumor regression at the end of the study) were previously 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), and age-matched untreated animals were inoculated subcutaneously in the left flank with EMT-6-MSA breast cancer cells (5x10 per mouse). 6 CT26 colon / colorectal cancer cells (3x10 cells per mouse) were inoculated subcutaneously on the right abdomen. 5 cells). Tumor growth was monitored and measured on both flanks twice a week, and animals were weighed twice a week.

[0338] Fig. 6A and 6BShown are the tumor volumes of mice previously treated with Conjugate 4 (0.3 / 0.012 mg / kg) when re-challenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells, respectively. Figure 6C and 6D The tumor volume of mice previously treated with conjugate 4 (0.30 / 0.012mg / kg) and anti-PD-1 RMP1-14 (5mg / kg) when challenged with EMT-6-MSA cells or CT26 colon / colorectal cancer cells is shown respectively. Mice previously treated with conjugate 4 or conjugate 4 and anti-PD-1 RMP1-14 (5mg / kg) were challenged with EMT-6-MSA cells again, resulting in 2 of 5 tumors being rejected and 4 of 4 tumors being rejected, respectively. Mice previously treated with conjugate 4 or conjugate 4 and anti-PD-1 RMP1-14 did not reject any cells when challenged with CT26 cells again. Mice treated with conjugate 4 as a monotherapy or conjugate 4 in combination with anti-PD-1 RMP1-14 showed immune memory.

[0339] Equivalent solution

[0340] The details of one or more embodiments of the present disclosure are set forth in the description attached above. Although any methods and materials similar or equivalent to those described herein can be used for the practice or testing of the present disclosure, preferred methods and materials are now described. Other features, objects and advantages of the present disclosure will be embodied in the specification and claims. In the specification and the appended claims, singular forms include plural references unless the context clearly provides otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present disclosure belongs. All patents and publications cited in this specification are incorporated by reference.

[0341] The foregoing description is presented for purposes of illustration only and is not intended to limit the present disclosure to the precise form disclosed except as limited by the appended claims.

Claims

1. A combination therapy comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy or at least one immunotherapy, wherein the STING agonist antibody-drug conjugate targeting HER2 is a conjugate of formula (A): wherein the conjugate comprises a HER2 antibody comprising a variable heavy chain complementary determining region 1 (CDRH1) comprising the amino acid sequence FTFSSYSMN (SEQ ID NO: 5); a variable heavy chain complementary determining region 2 (CDRH2) comprising the amino acid sequence YISSSSSTIYYADSVKG (SEQ ID NO: 6); a variable heavy chain complementary determining region 3 (CDRH3) comprising the amino acid sequence GGHGYFDL (SEQ ID NO: 7); and a variable light chain complementary determining region 1 (CDRL1) comprising the amino acid sequence RASQSVSSSY LA (SEQ ID NO: 12); a variable light chain complementary determining region 2 (CDRL2) comprising the amino acid sequence GASSRAT (SEQ ID NO: 13); and a variable light chain complementary determining region 3 (CDRL3) comprising the amino acid sequence QQYHHSPLT (SEQ ID NO: 14), and d 15 About 8.

2. The conjugate of claim 1, wherein the XMT-1519 antibody specifically binds to an epitope of the human HER2 receptor comprising residues 452 to 531 of the extracellular domain of the human HER2 receptor, residues 474 to 553 of SEQ ID NO: 1, or residues 452 to 531 of SEQ ID NO:

16.

3. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one HER2 targeted therapy.

4. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and at least one immunotherapy.

5. The combination of claim 1, wherein the HER2-targeted STING agonist antibody-drug conjugate enhances the efficacy of the HER2-targeted therapy or the immunotherapy.

6. The combination of claim 3, wherein the HER2 targeted therapy 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 HER2 small molecule inhibitor.

7. The combination of claim 6, wherein the antibody or antigen-binding fragment thereof that specifically binds to HER2 is a HER2 antibody, a HER2 dimerization inhibitor antibody, or a combination thereof.

8. The combination of claim 7, wherein the HER2 antibody or HER2 dimerization inhibitor antibody is trastuzumab, pertuzumab or magetuximab or a biosimilar thereof.

9. The combination of claim 6, wherein the HER2 targeted therapy is trastuzumab, pertuzumab, a combination of trastuzumab and pertuzumab, magetuximab, a trastuzumab biosimilar, a pertuzumab biosimilar, a combination of a trastuzumab biosimilar and a pertuzumab biosimilar, or a magetuximab biosimilar.

10. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and trastuzumab, pertuzumab, a combination of trastuzumab and pertuzumab, magetuximab, a trastuzumab biosimilar, a pertuzumab biosimilar, a combination of a trastuzumab biosimilar and a pertuzumab biosimilar, or a magetuximab biosimilar.

11. The combination of claim 6, wherein the HER2 targeting antibody-drug conjugate that specifically binds to HER2 is trastuzumab enalapril (T-DM1) or trastuzumab delutecan (trastuzumab-delutecan).

12. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and trastuzumab emtansine (T-DM1) or trastuzumab delutec (trastuzumab-delutec).

13. The combination of claim 6, wherein the HER2 small molecule inhibitor is tucatinib, neratinib or lapatinib.

14. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and tucatinib, neratinib or lapatinib.

15. The combination of claim 4, wherein the immunotherapy is an immune checkpoint inhibitor.

16. The combination of claim 15, wherein the immune checkpoint inhibitor is a monoclonal antibody, a humanized antibody, a fully human antibody, a fusion protein, or a combination thereof.

17. The combination of claim 15, wherein the immune checkpoint inhibitor is a PD-1 inhibitor or a PD-L1 inhibitor.

18. The combination of claim 17, wherein the PD-1 inhibitor or PD-L1 inhibitor is avelumab, durvalumab, dotarizumab, pembrolizumab, cemiplizumab, nivolumab, or atezolizumab.

19. The combination of claim 1, comprising at least one STING agonist antibody-drug conjugate targeting HER2 and avelumab, durvalumab, dotarizumab, pembrolizumab, cemiplizumab, nivolumab, or atezolizumab.

20. A method for treating, preventing, delaying progression of cancer or otherwise ameliorating symptoms of cancer in a subject, comprising administering the combination of claim 1.

21. The method of claim 20, 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, stomach cancer, or gastroesophageal junction cancer.

22. The method of claim 21, wherein the cancer is breast cancer, gastric cancer, gastroesophageal junction cancer, colorectal cancer, or non-small cell lung cancer.

23. The method of claim 20, wherein the immune checkpoint inhibitor and the conjugate are administered simultaneously.

24. The method of claim 20, wherein the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapy or the immunotherapy are administered simultaneously.

25. The method of claim 20, wherein the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapy or the immunotherapy are administered sequentially or alternately.

26. The method of claim 20, wherein the HER2-targeted STING agonist antibody-drug conjugate is administered prior to the HER2-targeted therapy or the immunotherapy.

27. The method of claim 20, wherein the HER2-targeted STING agonist antibody-drug conjugate is administered after the HER2-targeted therapy or the immunotherapy.

28. The combination of claim 1, wherein the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapy or the immunotherapy are in the same formulation.

29. The combination of claim 1, wherein the HER2-targeted STING agonist antibody-drug conjugate and the HER2-targeted therapy or the immunotherapy are formulated in separate formulations.

30. A kit comprising the combination of claim 1 and instructions for administration.

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