Methods for providing safe administration of Anti- CD40 antibodies

Abstract

To provide a method for clinically proven and safe intravenous delivery of anti- CD40 antibodies.SOLUTION: Intravenously administering to the subject a pharmaceutical composition comprising anti- CD40 antibodies and a pharmaceutically acceptable carrier, wherein the dosage is between 50 μ g / kg and 2500 μ g / kg, preferably between 75 μ g / kg and 2000 μ g / kg body weight 1kg of the subject per dose.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Reference to an electronically submitted sequence listing This application is filed under the file name "Sequence Li" created on May 22, 2019. sting_688097.0808" as an ASCII sequence table on EFS-Web The sequence listing, which is 10 kb in size, is submitted electronically via EFS-Web. The sequence listing submitted via the application is part of the present specification and is incorporated herein by reference in its entirety. be absorbed.

[0002] Background of the Invention CD40, a 48 kilodalton transmembrane cell-surface glycoprotein, inhibits tumor necrosis factor Cosminexin receptors belonging to the tumor necrosis factor receptor (TNFR) superfamily It is a stimulatory receptor (Elgueta R, et al. Immunol Rev., 200 9,229(1):152(-172). Constitutive expression of CD40 is variable and is expressed on dendritic cells. Antigen-presenting cells (antiDCs), including dendritic cells (DCs), B lymphocytes, and macrophages This receptor can be detected on the surface of antigen-presenting cells (APCs). CD40 is expressed on granulocytes, endothelial cells, smooth muscle cells, fibroblasts, and epithelial cells ( Korniluk,et al.TumourBiol.,2014,35(10):9 447-9457;Peters et al.Semin Immunol.,200 9,21(5):293-300).

[0003] Consistent with its widespread expression on normal cells, CD40 is also expressed in non-Hodgkin's lymphoma and hodgkin's lymphoma. Lymphoma, myeloma, and several tumors, including the nasopharynx, bladder, cervix, kidney, and ovary Present on the membranes of a wide range of malignant cells, including carcinomas (Eliopoulos AG & Youn g LS.,Curr.Opin.Pharmacol.,2004,4(4):360 CD40 interacts with a single ligand, CD40L (or CD154). , which is activated T lymphocytes, B lymphocytes, platelets, mast cells, macrophages, halophils Basal cells, natural killer (NK) cells, and non-hematopoietic cells (smooth muscle cells, It is a transmembrane protein expressed by cells (endothelial cells, and epithelial cells). As part of its action, CD40 binding to its only ligand, CD40L, mediates TNF receptor cleavage. Intracellular signaling pathways involve a series of adaptor molecules known as receptor activating factors (TRAFs). To initiate this intracellular signaling, multiple CD40 The receptors must form clusters on the cell membrane (Peters et al. Min Immunol., 2009, 21(5):293-300). Clustering to assemble supramolecular signaling complexes composed of multiple TRAFs This then leads to the activation of downstream transcription factors, including nuclear factor kappa B (NF-κB). (Kornbluth et al.Int.Rev.Immunol.,2012, 31(4):279-288).

[0004] The molecular consequences of CD40 signaling are determined by the cell types that express CD40 and the CD40 system. The signal depends on the microenvironment provided (Vonderheide et al. nCancer Res., 2013, 19(5):1035-1043). Irrigation and cross-linking stimulate the upregulation of membrane costimulatory molecules and MHC molecules and the pro-inflammatory cycle. Licensing adaptive immunity by APCs and especially DCs through induction of cytokine production CD40 is therefore essential for the functional maturation of APCs and, in turn, for the immune response. Involved in the activation of antigen-specific T lymphocytes (Long et al. Cancer Dis cov.,2016,6(4):400-13;Moran et al.Curr.O pin.Immunol.,2013,25(2):230-237). CD40 is also by activating resting B lymphocytes and increasing their antigen-presenting function. and plays a role in humoral immunity (Vonderheide et al. lin Cancer Res.,2013,19(5):1035-1043;Wol chok et al.Clin.Cancer Res.,2009,15(23): Furthermore, CD40 is expressed on NK cells, macrophages, and granulocytes. It is involved in the induction of innate immunity by stimulating cytotoxic myeloid cells (Rakhmile vich et al.Int.Rev.Immunol.,2012,31(4):2 67-278) The opposing roles of ATP in promoting both tumor progression and tumor cell apoptosis A large proportion of the disease has been attributed to the CD40 / CD450L pathway in various neoplastic diseases (Kor niluk et al.Tumour Biol.,2014,35(10):944 7-9457).

[0005] These important CD40 / CD40L-mediated pathways are essential for tumor progression in various neoplastic diseases. It has a dual role in both promoting proliferation and in inducing apoptosis of tumor cells ( Beatty GL,et al.Science,2011,331(6024):1 However, systemic administration of CD40 antibodies has been associated with shock syndrome and sarcopenia. associated with adverse side effects such as cytokine release syndrome (van Mierlo et al. al.,2002,Proc.Natl.Acad.Sci.USA,99:5561 -5566;van Mierlo et al.,2004,J Immunol 1 73:6753-6759). Examples of other anti-CD40 antibodies and their production are found in, e.g., This is described in US Patent No. 9,676,862.

[0006] In view of the above, improved anti-tumor therapies, particularly anti-CD40 agonists suitable for clinical use, are desired. There remains a need for antibodies.

[0007] Brief Summary of the Invention The present invention provides a method for administering anti-CD4 antibodies to a subject, including a clinically proven and safe treatment for advanced solid tumors. Regarding the clinically proven safe administration of 0 antibodies.

[0008] In one general aspect, the present invention provides for the clinical administration of anti-CD40 antibodies to human subjects in need thereof. 10. A method for providing clinically proven safe administration of an antibody and a pharmaceutically acceptable carrier. The method comprises intravenously administering to a subject a pharmaceutical composition comprising a carrier containing the antibody, and preferably the antibody is The heavy chain variable region is represented by SEQ ID NOs: 1, 2, and 3, respectively. Heavy chain complementarity determining region (HCDR) of The amino acid sequences of HCDR1, HCDR2, and HCDR3, or SEQ ID NOs: 1 and 2, respectively , and 3 and at least 95% (or at least 96%, at least 97%, at least 9 8%, or at least 99%) sequence identity, and the light chain variable regions are The light chain complementarity determining regions of SEQ ID NOs: 4, 5, and 6 amino acid sequences of LCDR1, LCDR2, and LCDR3; At least 95% (or at least 96%, at least) identical to SEQ ID NOs: 4, 5, and 6, respectively at least 97%, at least 98%, or at least 99%) sequence identity, The total dose of antibody administered is 50 μg / kg of subject body weight per administration. g to 2500 μg / kg, preferably 75 μg to 2000 μg / kg, optionally 100 μg / kg~1800μg / kg, 200μg / kg~1500μg / kg, 300μg / kg~1400μg / kg, 400μg / kg~1300μg / kg, 500μg / k g~1200μg / kg, 600μg / kg~1100μg / kg, 700~1000μ g / kg, 800 to 900 μg / kg.

[0009] In one embodiment, the human subject has been diagnosed with an advanced solid tumor.

[0010] In one embodiment, the anti-CD40 antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO:7. (VH) and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO:8.

[0011] In one embodiment, the anti-CD40 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO:9. and a light chain (LC) having the amino acid sequence of SEQ ID NO: 10. nothing.

[0012] In some embodiments, the total dose of anti-CD40 antibody administered per administration is Per kg of subject body weight, 50 μg / kg, 75 μg / kg, 100 μg / kg, 200 μg / kg, 300μg / kg, 400μg / kg, 500μg / kg, 600μg / k g, 700μg / kg, 800μg / kg, 900μg / kg, 1000μg / kg, 1 100μg / kg, 1200μg / kg, 1300μg / kg, 1400μg / kg, 1 500μg / kg, 1800μg / kg, 2000μg / kg, or 2500μg / kg or any dose therebetween.

[0013] In one embodiment, the total administration time of the pharmaceutical composition is 0 minutes to 3 hours, preferably 5 minutes to 150 minutes. , 10 minutes to 2 hours, 15 minutes to 90 minutes, 20 minutes to 1 hour, 25 minutes to 55 minutes, 30 minutes to 50 minutes , or 35 to 45 minutes, or 40 to 45 minutes, for example, about 0 minutes, about 5 minutes, about 10 minutes , about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes , about 55 minutes, about 1 hour, about 90 minutes, about 2 hours, about 150 minutes, or about 3 hours The pharmaceutical composition is administered intravenously to the subject. Preferably, the pharmaceutical composition is administered daily, weekly, monthly, six months, or Repeatedly, i.e., more than once, over a period of one, two or more years, more preferably Once a day, once a week, once every two weeks, once a month, once every six months, etc. It is administered intravenously to the subject.

[0014] In one embodiment, the method comprises administering a therapeutic agent before, after, or simultaneously with the administration of an anti-CD40 antibody. to a human subject, preferably wherein the therapeutic agent is a corticosteroid, an anti-inflammatory drug, Selected against the group consisting of histaminics, antipyretics, H2-antagonists, and antiemetics do.

[0015] In one embodiment, the pharmaceutical composition comprises 1 mg / mL to 100 mg / mL of an anti-CD40 antibody, Preferably, 10 mg / mL to 90 mg / mL, 20 mg / mL to 80 mg / mL, 30 mg g / mL~70mg / mL, 40mg / mL~60mg / mL, 40mg / mL~50m g / mL, e.g., 10 mg / mL, 20 mg / mL, 30 mg / mL, 40 mg / mL , 50mg / mL, 60mg / mL, 70mg / mL, 80mg / mL, 90mg / mL or 100 mg / mL of anti-CD40 antibody and a pharmaceutically acceptable carrier.

[0016] In another general aspect, the invention provides a method for the clinical administration of an anti-CD40 antibody to a human subject in need thereof. 10. A method for providing clinically proven safe administration of an antibody and a pharmaceutically acceptable carrier. The method comprises intravenously administering to a subject a pharmaceutical composition comprising a carrier containing the antibody, and preferably the antibody is The heavy chain variable region is represented by SEQ ID NOs: 1, 2, and 3, respectively. Amino acids of heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3 of or at least 95% (or at least 96%, Sequences with at least 97%, at least 98%, or at least 99% sequence identity and the light chain variable region comprises the light chain complementarity determining regions (LCs) of SEQ ID NOs: 4, 5, and 6, respectively. DR) Amino acid sequences of LCDR1, LCDR2, and LCDR3 or SEQ ID NO: 4, respectively , 5, and 6 and at least 95% (or at least 96%, at least 97%, at least and the total number of antibodies administered contains a sequence with at least 98%, or at least 99%, sequence identity to the target gene. The dosage is approximately 600 μg / kg to 900 μg per kg of subject body weight per administration. / kg, and preferably the human subject has non-small cell lung cancer The present invention relates to a method for treating a patient diagnosed with NSCLC, pancreatic cancer, or cutaneous melanoma.

[0017] In one embodiment, the anti-CD40 antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO:7. (VH) and a light chain variable region (VL) having the amino acid sequence of SEQ ID NO:8.

[0018] In one embodiment, the anti-CD40 antibody has a heavy chain (HC) having the amino acid sequence of SEQ ID NO: 9. and a light chain (LC) having the amino acid sequence of SEQ ID NO: 10.

[0019] In one embodiment of the present invention, a compound as described in the previous embodiment for use in medicine is provided. A suitable anti-CD40 antibody is provided.

[0020] In a further embodiment of the present invention, the present invention is directed to the treatment of bladder cancer, breast cancer, cervical cancer, colon cancer, endometrial cancer, kidney cancer, , oral cancer, liver cancer, melanoma (including cutaneous melanoma), mesothelioma, non-small cell lung cancer, non-melanoma skin cancer, including but not limited to oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, small cell lung cancer, and thyroid cancer

[0032] The present invention also includes, but is not limited to, the use of the compounds described in the preceding embodiments for use in the treatment of advanced solid tumors. Such anti-CD40 antibodies are provided.

[0021] In a further embodiment of the present invention, the present invention is directed to the treatment of bladder cancer, breast cancer, cervical cancer, colon cancer, endometrial cancer, kidney cancer, , oral cancer, liver cancer, melanoma (including cutaneous melanoma), mesothelioma, non-small cell lung cancer, non-melanoma skin cancer, including but not limited to oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, small cell lung cancer, and thyroid cancer any of the foregoing for use in the manufacture of a medicament for the treatment of advanced solid tumors, including, but not limited to, The present invention provides an anti-CD40 antibody as described in the embodiments above.

[0022] Preferably, in these two embodiments, the treatment of advanced solid tumors comprises the use of antibodies and pharmaceuticals. and administering to a subject a pharmaceutical composition comprising a carrier acceptable for the method of the present invention intravenously to the subject. administering an anti-CD40 antibody as defined above to a human subject diagnosed with The total dose of antibody administered is 50 μg / kg of subject body weight per administration. to 2500 μg / kg, preferably 75 μg / kg to 2000 μg / kg.

[0023] The details of one or more embodiments of the invention are set forth in the description below. Other features and advantages These points will become apparent from the following detailed description and the appended claims. [Brief explanation of the drawings]

[0024] The above Summary of the Invention and the following Detailed Description of the Invention should be read in conjunction with the accompanying drawings. The invention will be better understood by reading the accompanying drawings, in which: It should be understood that the present invention is not limited to the above.

[0025] This patent or application file contains at least one color-printed drawing. Copies of this patent or patent application publication are available upon request and payment of the necessary fee. This will be provided by the Patent Office. [Figure 1]1 is a diagrammatic representation of the study design for the clinical trial in Example 1. IV = intravenous, NSCLC = non-small cell lung cancer, q14d = every 14 days, RP2D = recommended Phase II dose. [Figure 2] FIG. 1 shows the study design and cohorts with and without corticosteroid pre-infusion. [Figure 3] Graph of incidence of infusion-related reactions (IRRs) per assigned dose, censored by dose escalation. [Figure 4] Graph showing mean serum concentrations over time cycles 1 and 2. [Figure 5] 1 is a graph showing dose-normalized AUC0-24h. [Figure 6] A-C are graphs showing the percentage of B cells (A), T cells (B), and NK cells (C) in peripheral blood after infusion of antibody A, normalized to pre-infusion levels (no corticosteroid cohort). [Figure 7] A-I are graphs showing cytokine / chemokine levels in peripheral blood after infusion of antibody A (no corticosteroid cohort): MCP-1 (A), IP-10 (B), MIP-1β (C), IFN-γ (D), MIP-1α (E), IL-8 (F), TNF-α (G), IL-6 (H), and IL12p70 (I). [Figure 8] A-D are graphs showing the expression of activation / maturation markers in peripheral blood B lymphocytes: HLA-DR (A), CD54 (B), CD80 (C), and CD86 (D). Symbols and lines represent each individual patient from Cohort 6B expansion (see Figure 2). Note: The fold change in staining intensity (24 h post-infusion of antibody A vs. pre-infusion) was calculated for each marker and converted to the Log2 scale. Of the six patients in the final cohort (1200 μg / kg without corticosteroids), four patients had usable data and were graphed accordingly.

[0026] Detailed Description of the Invention Various publications, articles and patents are cited or referenced in the Background and throughout this specification. Each of these references is incorporated herein by reference in its entirety. Any discussion of documents, operations, materials, devices, articles or the like which is included in the specification is indicative of the context of this invention. Such discussion is intended to provide a basis for understanding whether any or all of these matters are disclosed or implied. is admitted to be part of the prior art to any claimed invention. isn't it.

[0027] Unless otherwise defined, all technical and scientific terms used herein refer to the It has the same meaning as commonly understood by one of ordinary skill in the art to which it pertains. Certain terms used herein have the meanings set forth herein. All patents, published patent applications and publications cited herein are hereby incorporated by reference. No. 6,239,793, filed on Oct. 1, 2004, which is incorporated in its entirety as if set forth herein.

[0028] As used in this specification and the appended claims, the singular forms "a," "an," and " It should be noted that "the" includes multiple referents unless the context makes clear otherwise. There is.

[0029] Unless otherwise indicated, the term "at least" preceding a series of elements refers to every element in the series. It should be understood that the present invention refers to elements of the present invention. and recognize many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by this invention. is intended.

[0030] Throughout this specification and the claims that follow, unless the context otherwise requires, the term "comprises" "comprises" and variations such as "comprises" and "comprising" are used to refer to Including the specified integer or step or group of integers or steps, but any other integer or step does not mean to exclude any integer or step or group of steps. It will be understood that, as used herein, the term "comprising" includes the term " "containing" or "including" can be substituted, or When used herein, the term "having" may be substituted.

[0031] As used herein, "consisting of" refers to a claim element. The term "compound" as used herein excludes any element, step, or ingredient not specified therein. When the term "consisting essentially of" is used, The present invention does not exclude materials or steps that do not materially affect the present invention's novel characteristics. When used herein in connection with an aspect or embodiment, it is intended to vary the scope of the disclosure. "comprising," "containing," "including," and Any of the above terms "comprising" and "having" may be replaced with the terms "consisting of" or "consisting essentially of." can be replaced with

[0032] As used herein, the conjunctive term "and / or" between multiple listed elements means It is understood to encompass both individual and combined options. For example, When elements are connected by "and / or," the first option may be selected without the second option. The first option indicates that the second option is applicable without the first option. The third option is that the first and second elements are applicable together. Any one of these options is included in the meaning and therefore As used in the specification, it is understood that the requirements of the term "and / or" are met. The simultaneous applicability of two or more of the following is also included in the meaning, and therefore the term "and / or" It is understood that the requirements of

[0033] As used herein, the term "subject" refers to a subject who is administered an anti-CD40 antibody according to the methods of the present invention. mammalian subjects, preferably mammalian subjects, who have been diagnosed with or are being administered a Diagnosis of cancer may be performed by a clinical diagnostic test, physical examination of the subject, or by the diagnosis of a specific disease. or by a clinician according to any other accepted method for diagnosing a subject with a disease. This can be done.

[0034] As used herein, CD40 refers to a member of the tumor necrosis factor receptor (TNFR) superfamily. It is a glycoprotein expressed on the cell surface that belongs to the genus Lysosome and plays a central role in the immune system. CD40 is expressed on various immune cells, such as B cells, dendritic cells, monocytes, and macrophages. and professional APCs signal through CD40. It is activated when (Reviewed by J. Sci., 2001, (58):4-43). CD40 expression It is expressed in many normal cells and tumor cells, including B lymphomas, solid tumors, melanomas, and carcinomas. Activation of CD40 is effective in inducing antitumor responses, and CD40 activation This suggests that CD40-positive tumors may be involved in at least one of the immune activation mechanisms, including a direct apoptotic effect on CD40-positive tumors. , as well as antibody-dependent cell-mediated cytotoxicity icity, ADCC) and complement dependent cytotoxicity, C Stimulation of humoral responses that trigger DCs may contribute to impaired tumor growth. It is established.

[0035] As used herein, "CD40" refers to the human CD40 protein as defined herein. have structural and / or functional identity to proteins and / or their natural variants Preferably, CD40 is a protein derived from a Unicellular or human host cell. Prot accession number P25942 and GenBank accession number AAH and human CD40 such as 12419.

[0036] As used herein, an "anti-CD40 antibody" refers to an antibody that binds to the natural ligand CD40L. and an agonistic human monoclonal antibody of the IgG1 subtype that enhances the effect of IgG1. The term "agonist" refers to an antibody or a fragment thereof, such as a human clonal antibody (mAb), or an antigen-binding fragment thereof. Anti-CD40 antibodies (1) directly bind to the CD40 receptor expressed on tumor cells; (2) induces a direct antitumor effect, and (3) promotes the "licensing" of DCs and the activation of cytotoxic T cells (cytotoxic T cells). (3) activation of cytotoxic T-cells (CTLs), and (4) activation of cytotoxic myeloid cells such as NK cells or may induce an indirect antitumor effect by activating tumor macrophages. In one embodiment, the anti-CD40 antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises: Heavy chain complementarity determining regions (HCDRs) HCDR1, HCD of SEQ ID NOs: 1, 2, and 3, respectively HCDR2, and HCDR3 or at least the amino acid sequences of SEQ ID NOs: 1, 2, and 3, respectively 95% (or at least 96%, at least 97%, at least 98%, or less The light chain variable regions contain sequences with 99% sequence identity to each other, and the light chain variable regions contain sequences with 99% sequence identity to each other, respectively. and 6 light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3. or SEQ ID NOs: 4, 5, and 6, respectively, with at least 95% (or at least 96%) %, at least 97%, at least 98%, or at least 99%) of sequence identity In another preferred embodiment, the anti-CD40 antibody comprises the amino acid sequence of SEQ ID NO:7. a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO: 8 and a light chain variable region (V In another preferred embodiment, the anti-CD40 antibody comprises the amino acid sequence of SEQ ID NO: 9. and a light chain (LC) having the amino acid sequence of SEQ ID NO: 10.

[0037] Additional anti-CD40 antibodies or antigen-binding fragments thereof that can be used in the present invention include , as described in U.S. Pat. No. 9,676,862, which is incorporated herein by reference. Includes things.

[0038] Anti-CD40 antibodies can be produced by monoclonal antibodies, including but not limited to hybridoma production. by any method known in the art in light of the present disclosure for preparing monoclonal antibodies. For example, anti-CD40 antibodies can be prepared using recombinant DNA technology. , mammalian cell lines (e.g., Chinese Hamster Ovary, CH O) cell lines). In particular, the generation of anti-CD40 antibodies useful in the present invention Methods are described, for example, in U.S. Pat. No. 9,676,862, which is incorporated herein by reference. No. 6,299,499.

[0039] The term "safe" refers to a dose, administration regimen, treatment, or method using an anti-CD40 antibody. In cases involving: Treatment-emergent adverse events (A) compared with another comparator according to the Cosmetic Act acceptable frequency and / or severity of adverse events (termed TEAEs or TEAEs) refers to the risk:benefit ratio (secs. 201-902, 52 Stat. 104 0 et seq., as amended;21U.SC§§321-392). In particular, the safety associated with the dose, administration regimen, or treatment with the anti-CD40 antibodies of the invention may be determined by: Adverse events were considered to be possibly, likely, or very likely due to the use of anti-CD40 antibodies. If possible, the acceptable frequency and / or tolerability of adverse events associated with antibody administration Safety often refers to the minimum amount of an active pharmaceutical ingredient needed to achieve the desired effect. This is measured by toxicity studies to determine the maximum tolerated dose or optimal dose. It also seeks to identify any potential adverse effects that exposure to the drug may have. be.

[0040] As used herein, unless otherwise specified, the term "clinically proven" (independently or used to modify the term "safe") is proven by clinical trials Clinical trials are being conducted under the supervision of the U.S. Food and Drug Administration and the European Medicines Evaluation Agency. This means that the product meets the approval criteria of the European Union (EU) or the corresponding national regulatory body. In the present invention, clinical trials are conducted in patients with advanced solid tumors. Safety and efficacy of Antibody A, an agonistic human monoclonal antibody targeting CD40 This is a phase I, open-label study of pharmacokinetics and pharmacodynamics.

[0041] As used herein, the terms "adverse event," "treatment-emergent adverse event," and "treatment-emergent adverse event" are used interchangeably. An "adverse reaction" is any adverse event associated with or resulting from the administration of a pharmaceutical composition or therapeutic agent. Means an adverse, unfavourable, unintended or undesirable symptom or outcome. However, abnormal values ​​or observations were not deemed clinically significant by the investigator. As used herein, an adverse event is not reported unless the event is In this case, "clinically evident" means the clinically evident or apparent to a physician or clinical investigator, using criteria acceptable to those skilled in the art. Clinically significant as determined by the investigator. When adverse or undesirable consequences reach such a level of severity, regulatory authorities may In some cases, a pharmaceutical composition or therapeutic agent may be deemed unacceptable for its proposed use. Examples of adverse events or reactions when CD40 antibodies are used in the context of intravenous administration include: Symptoms include, but are not limited to: rhinitis, shingles, and bullous tympanocystitis infections and infestations; respiratory, thoracic, and mediastinal disorders such as cough, pharyngeal irritation, and sore throat; diarrhea and Gastrointestinal disorders such as bloating and flatulence; nervous system disorders such as headaches and dizziness; anemia and lymphadenopathy Blood and lymphatic system disorders; back pain, premature labor, infusion reactions, local injection site reactivity, malignancy, and Anaphylaxis or serum sickness-type reaction.

[0042] As used herein, "treatment" or "treating" refers to therapeutic treatment. Individuals of interest include subjects diagnosed with a disorder or symptoms of a disorder. Subjects who can be treated include those prone to having the disorder or those susceptible to the disorder. Among the things that have the property, those that can prevent such disorders can also be mentioned. Desired clinical outcomes include symptomatic relief, disease progression, whether detectable or undetectable. reduction in severity of disease, stable (i.e., not worsening) disease state, delay or slowing of disease progression These include improvement or alleviation of the disease state, and remission (whether partial or total). Beneficial clinical outcomes include, for example, increased B cell or dendritic cell proliferation in treated subjects. Reduced proliferation of inflammatory cytokines, adhesion molecules, proteases, immunoglobulins, The combination of these factors reduces the production of anti-inflammatory proteins, reduces the number of autoreactive cells, Enhancement of immune tolerance, inhibition of survival of autoreactive cells, and / or CD40 / CD40L-mediated pathways Clinical responses include a reduction in one or more symptoms mediated by magnetic resonance imaging (MRI). magnetic resonance imaging (MRI) scan, X-ray fluoroscopy, and computed tomography ( Computed tomography (CT) scan, flow cytometry, or fluorescence-activated cell microscopy fluorescence-activated cell sorter (FACS) analysis, tissue diagnosis, macroscopic findings, and and changes detectable by ELISA, RIA, and chromatography, etc. The presence of steroids can be assessed using screening techniques, such as, but not limited to, blood chemistry. do.

[0043] The terms "efficacious" and "effective" in the context of a dosage, administration regimen, treatment, or method As used herein, the term "effectiveness" refers to the effectiveness of a particular dosage, administration, or treatment regimen. Efficacy can be measured based on changes in the course of a disease in response to an agent of the invention. For example, an anti-CD40 antibody of the invention (e.g., Antibody A) may be used to measure the severity of the disorder being treated. sufficient to cause an improvement, preferably a sustained improvement, in at least one indicator of The amount and duration of the treatment is determined to be sufficient. To assess the severity of a disease, disorder, or condition in a subject, various indicators that reflect the severity of the disease, disorder, or condition can be evaluated. Indicators of interest include, for example, clinically recognized disease severity, symptoms, or manifestations of a subject's disorder. The degree of improvement is assessed overall by the doctor, who then reports this assessment to the patient. can be based on signs, symptoms, biopsy, or other test results and can be administered to a subject Employ questionnaires, such as quality of life questionnaires developed for a given disease. For example, the anti-CD40 antibodies of the present invention can be used to treat advanced solid tumors. It may be administered to achieve improvement in the condition.

[0044] Disease assessment for advanced solid tumors requires computed tomography (CT) or These include magnetic resonance imaging (MRI) and bone scans for subjects with prostate cancer. Disease response was assessed according to the Response Evaluation Criteria in Solid Tumors (RESE). According to RECIST v1.1 and immune-related response criteria (Immune-Related Response Criteria (IRC) v1.1), The study will be evaluated according to the International Response Criteria (IRRC). Uses the Prostate Cancer Clinical Trials Working Group (PCWG3) criteria35 and assessing disease response in subjects with prostate cancer.

[0045] As used herein, an advanced solid tumor is one that has spread widely to other anatomical sites. As used herein, accumulation refers to a malignant solid neoplasm that has progressed or is no longer responsive to treatment. The product is measured by pharmacokinetic measurements that show accumulation and increase with repeated administration of anti-CD40 antibody. This refers to the amount of anti-CD40 antibodies in the blood.

[0046] As used herein, the dose of an anti-CD40 antibody in "μg / kg" refers to the amount of the antibody administered. The amount of anti-CD40 antibody administered in micrograms per kilogram of subject body weight Point.

[0047] In one general aspect, the present invention provides a method for treating a subject, preferably a human subject, in need thereof. The present invention relates to a method for providing clinically proven and safe intravenous administration of anti-CD40 antibodies. Preferably, the subject has any type of progressive or refractory solid malignancy that is metastatic or unresectable. Examples of the above diseases include bladder cancer, breast cancer, cervical cancer, colon cancer, Endometrial cancer, kidney cancer, oral cancer, liver cancer, melanoma (including cutaneous melanoma), mesothelioma, non-small cell Lung cancer, non-melanoma skin cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, small cell lung cancer, and thyroid cancer These include, but are not limited to, esophageal adenocarcinoma.

[0048] In one embodiment, the clinically proven safe administration and / or administration of an anti-CD40 antibody to a subject Alternatively, a method for providing safe treatment of an advanced solid tumor in a subject, preferably a human subject, comprising: A pharmaceutical composition comprising an anti-CD40 antibody and a pharmaceutically acceptable carrier is intravenously administered to a subject. The total dose of the anti-CD40 antibody administered is 1 kg of the subject's body weight per administration. 50 μg / kg to 2500 μg / kg, preferably 75 μg / kg to 2000 μ g / kg mg / kg, optionally 100 μg / kg to 1800 μg / kg, 200 μg / kg~1500μg / kg, 300μg / kg~1400μg / kg, 400μg / kg~1300μg / kg, 500μg / kg~1200μg / kg, 600μg / k 100μg / kg, 700μg / kg, 800μg / kg, 1100μg / kg, 700μg / kg, 1000μg / kg, 800μg / kg do.

[0049] Intravenous administration refers to administration directly into a vein. Intravenous administration is by injection (e.g. by injection (e.g., using a syringe at higher pressure) or by injection (e.g., using gravity pressure). Intravenous administration typically allows a drug or therapeutic agent to be delivered through the blood circulation. It is the fastest way to deliver drugs or therapeutic agents throughout the body. When administration of the antibody is via intravenous administration, it may be administered by intravenous infusion or injection; Preferably, the total amount of anti-CD40 antibody administered to a subject per administration is 100 mg / kg. The dose is administered over approximately 30 minutes, such as 30, 60, 90, 120, 150, or 180 minutes. for a period of 1 to 180 minutes, preferably 60 to 120 minutes, optionally 90 to 120 minutes. Optionally, the total dose can be administered within 0 to 3 hours. Preferably, 5 to 150 minutes, 10 to 2 hours, 15 to 90 minutes, 20 to 1 hour, 2 Duration of 5 to 55 minutes, 30 to 50 minutes, 35 to 45 minutes, or 40 to 45 minutes, e.g. For example, about 0 minutes, about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, Approximately 40 minutes, approximately 45 minutes, approximately 50 minutes, approximately 55 minutes, approximately 1 hour, approximately 90 minutes, approximately 2 hours, approximately 150 minutes or by intravenous infusion over a period of about 3 hours.

[0050] The total dose of anti-CD40 antibody per administration was determined intravenously in clinical trials. The administration is selected to provide safe administration and / or safe treatment. According to the embodiment, when the pharmaceutical composition is administered intravenously, the amount of anti-CD14 antibody administered per administration is The total dose of the antibody may be, for example, 50 μg / kg, 75 μg / kg, 100 μg / kg, 200μg / kg, 300μg / kg, 400μg / kg, 500μg / kg, 600μ g / kg, 700μg / kg, 800μg / kg, 900μg / kg, 1000μg / k g, 1100μg / kg, 1200μg / kg, 1300μg / kg, 1400μg / k g, 1500 μg / kg, 1800 μg / kg, or 2000 μg / kg, or Any dose between these.

[0051] The total dose of anti-CD40 antibody may be administered daily, weekly, monthly, six months, one year, two years, or longer. Over the above period, once a day, once a week, once every two weeks, once a month, once every six months For example, 75 μg / kg to 2000 μg / kg of antibody can be administered once a day. The total dose of CD40 antibody is administered by a single intravenous injection, with the dose being 200 mg / kg or less per administration (e.g., 100 mg / kg or less). at least once a day), i.e., 0 minutes to 3 hours, preferably 5 minutes to 150 minutes, 10 minutes to 2 hours, 15 minutes to 90 minutes, 20 minutes to 1 hour, 25 minutes to 55 minutes, 30 minutes to 50 minutes, Or 35 to 45 minutes, or 40 to 45 minutes, for example, about 0 minutes, about 5 minutes, about 10 minutes, Approximately 15 minutes, approximately 20 minutes, approximately 25 minutes, approximately 30 minutes, approximately 35 minutes, approximately 40 minutes, approximately 45 minutes, approximately 50 minutes, It may be administered for about 55 minutes, about 1 hour, about 90 minutes, about 2 hours, about 150 minutes, or about 3 hours. Multiple anti-CD40 antibodies at total doses of 75 μg / kg to 2000 μg / kg each were administered. A single dose can be administered to a subject in need thereof.

[0052] Pharmaceutical compositions suitable for use in the methods of the present invention are formulated for intravenous administration. Examples of formulations suitable for oral administration include solutions, suspensions, emulsions, and pharmaceutical preparations for injection or infusion. These include, but are not limited to, dry products that can be dissolved or suspended in an acceptable carrier. In a preferred embodiment, a pharmaceutical composition comprising an anti-CD40 antibody for use in the methods of the present invention is provided. The composition is formulated as a liquid.

[0053] The concentration of anti-CD40 antibody contained in the pharmaceutical compositions used in the present invention may vary. Typically, the concentration of the anti-CD40 antibody is 1 mg / mL to 100 mg / mL, preferably , 10mg / mL~90mg / mL, 20mg / mL~80mg / mL, 30mg / mL ~70mg / mL, 40mg / mL~60mg / mL, 40mg / mL~50mg / mL For example, 1 mg / mL, 10 mg / mL, 20 mg / mL, 30 mg / mL, 4 0mg / mL, 50mg / mL, 60mg / mL, 70mg / mL, 80mg / mL, 9 0 mg / mL, or 100 mg / mL, or any concentration therebetween. In this form, the concentration of the anti-CD40 antibody is 10 mg / mL to 30 mg / mL, for example, 20 mg / mL. In one embodiment, preferably the concentration of the anti-CD40 antibody is 20 mg / m The concentration is 1 to 60 mg / mL, for example, 40 mg / mL.

[0054] Pharmaceutical compositions for use in the present invention may comprise one or more pharmaceutically acceptable carriers, e.g., It also includes those widely used in the field of pharmaceutical production, particularly antibody pharmaceutical production. As used herein, the term "carrier" refers to any excipient, diluent, buffer, stabilizer, or pharmaceutical Other materials well known in the art for formulation. In particular, pharmaceutically acceptable carriers are non-toxic. and must not interfere with the efficacy of the active ingredient. Examples include excipients and / or esters suitable for use in pharmaceutical compositions known in the art. Examples of additives include those described in "Re" (Reaction Ingredients), the entire disclosure of which is incorporated herein by reference. minton:The Science&Practice of Pharmacy ”19th ed., Williams&Williams, (1995)” and “Ph. ysician's Desk Reference" 52nd ed., Medica 1 Economics, Montvale, NJ (1998) .

[0055] According to an embodiment of the invention, the pharmaceutical composition for use in the invention comprises an anti-CD40 antibody and and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier is , one or more amino acids such as L-histidine and / or glycine, lactose, maltose One or more carbohydrates such as sucrose, trehalose, polysorbate 20, polysorbate one or more surfactants such as sorbate 80, and one or more alcohols such as D-sorbitol Preferably, the pharmaceutical composition contains 5 to 6, preferably 5.1 to 5.9, 5. pH between 2 and 5.8, 5.3 and 5.7, 5.4 and 5.6, and 5.4 and 5.5, e.g., 5.0 , 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, Young has a pH of 6.0, or any value therebetween.

[0056] In some embodiments, the pharmaceutical compositions for use in the present invention comprise L-histidine. and one or more amino acids such as glycine. The amino acids may be present in an amount of 1 mM to 40 mM, 1 mM ~20mM, 5mM~35mM, 10mM~30mM, 15mM~25mM, or 2 0 mM to 40 mM, or 0.50% to 2.00% (weight / volume) (w / v), preferably 0.75%(w / v)~1.75%(w / v), 1.00%(w / v)~1.50%(w It can be present at a concentration of 1.00% (w / v) or 1.25% (w / v). For example, the pharmaceutical composition may be at 1 mM, 5 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM L-histidine or glutathione at 10 mM, 35 mM, or 40 mM, or any concentration therebetween. In another example, the pharmaceutical composition may contain 0.50% (w / v), 0.75% (w / v) lysine. / v), 1.00%(w / v), 1.25%(w / v), 1.50%(w / v), 1.7 L-His at 5% (w / v), or 2.00% (w / v), or any concentration therebetween. It may contain stigmine or glycine.

[0057] In some embodiments, the pharmaceutical compositions for use in the present invention contain 1% to 10% (by weight) / volume)(w / v), 5%-10%(w / v), 8%-9%(w / v), 1.5%(w / v)~9.5%(w / v), 2%(w / v)~9%(w / v), 2.5%(w / v)~8 .5%(w / v), 3%(w / v)~8%(w / v), 3.5%(w / v)~7.5%( w / v), 4%(w / v)~7%(w / v), 4.5%(w / v)~6.5%(w / v) , at a concentration of 5% (w / v) to 6% (w / v), or 5% (w / v) to 5.5% (w / v) at least one sugar, such as sucrose, glucose, cellobiose, or trehalose; For example, the pharmaceutical composition may contain 1% (w / v), 1.5% (w / v), 2% (w / v) , 2.5%(w / v), 3%(w / v), 3.5%(w / v), 4%(w / v), 4.5 %(w / v), 5%(w / v), 5.5%(w / v), 6%(w / v), 6.5%(w / v), 7%(w / v), 7.5%(w / v), 8%(w / v), 8.5%(w / v), 9 %(w / v), 9.5%(w / v), or 10%(w / v), or any value therebetween The composition may contain sucrose, cellobiose, and / or trehalose at a concentration of

[0058] In some embodiments, pharmaceutical compositions for use in the present invention contain 0.01% (w / v )~0.10%(w / v), 0.01%(w / v)~0.08%(w / v), 0.02% (w / v)~0.05%(w / v), 0.02%(w / v)~0.09%(w / v), 0 .03%(w / v)~0.08%(w / v), 0.04%(w / v)~0.07%(w / v), or polysorbate 80 ( at least one surfactant such as polysorbate 20 (PS80) or polysorbate 20 (PS20) For example, the concentration of polysorbate 20 and / or polysorbate 80 is 0.01% ( w / v), 0.02%(w / v), 0.03%(w / v), 0.04%(w / v), 0. 05%(w / v), 0.06%(w / v), 0.07%(w / v), 0.08%(w / v ), 0.09% (w / v), or 0.1% (w / v), or any concentration therebetween may be.

[0059] In some embodiments, pharmaceutical compositions for use in the present invention contain 0.01% (w / v )~0.10%(w / v), 0.01%(w / v)~0.08%(w / v), 0.02% (w / v)~0.05%(w / v), 0.02%(w / v)~0.09%(w / v), 0 .03%(w / v)~0.08%(w / v), 0.04%(w / v)~0.07%(w / v), or mannitol, xylitol, at a concentration of 0.05% (w / v) to 0.06% (w / v). It contains at least one polyol, such as maltodextrin, thiamin, or D-sorbitol. The concentrations of ethanol, xylitol, or D-sorbitol were 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09 %, or 0.1% (w / v), or any concentration therebetween.

[0060] Pharmaceutical compositions comprising anti-CD40 antibodies for use in the present invention may be prepared by methods known in the art in light of this disclosure. It can be prepared by any method known in the art. For example, an anti-CD40 antibody can be mixed with one or more pharmaceutically acceptable carriers to obtain a solution. Until administration to the subject, the temperature should be kept between -40°C ± 10°C and -70°C ± 20°C (minus 40°C ± 10°C at a controlled temperature in the range of -70°C ± 20°C and in suitable vials. It can be stored as a frozen liquid, protected from light.

[0061] According to embodiments of the present invention, supportive care, such as pre-infusion and post-infusion supportive care, is administered to patients receiving anti-CD40 In addition to administering the antibody, it may be used to treat advanced solid tumors. The human subject receives a pre-infusion medication prior to administration of the anti-CD40 antibody. The human subject receives a post-infusion medication following administration of the anti-CD40 antibody. In some embodiments, Human subjects receive supportive care medications concurrently with administration of the anti-CD40 antibody. Examples of these medications include: These include corticosteroids, antihistamines, antipyretics, H2-antagonists, and antiemetics. Medications include, but are not limited to, corticosteroids, Receive all antihistamines, antipyretics, H2-antagonists, and antiemetics. Typically, subjects receive antihistamines, antipyretics, H2-antagonists, and antiemetics. Optionally, the subject is also receiving corticosteroids, antihistamines, antipyretics, H2-antagonists, Patients will receive one or more of the following: antiemetics, anticoagulants, and antiemetics.

[0062] Examples of suitable corticosteroids include dexamethasone and methylprednisolone. Dexamethasone is administered at 20 mg and methylprednisolone at 80 mg. Examples of suitable antihistamines include diphenylhydramine and cetyl Cetirizine can be administered in 10 mg doses. Examples of suitable antipyretics include: Examples of antihistamines include acetaminophen (paracetamol). , which can be administered at a dose of 650 to 1000 mg. Examples of suitable H2-antagonists include Examples of suitable antiemetics include ranitidine, which may be administered in 50 mg doses. An example is ondansetron, which can be administered in 8 mg doses. .

[0063] According to embodiments of the present invention, various factors may be analyzed to determine clinical outcomes, such as those described herein. Studies will determine whether specific doses of anti-CD40 antibodies provide safe intravenous administration. For example, the safety of a particular dose of an intravenously administered anti-CD40 antibody can be determined. The efficacy of the antibody can be assessed by immunogenicity testing (e.g., measuring the production of antibodies against anti-CD40 antibodies). This will determine dose-limiting toxicity (DLT) in subjects. Thus, serum proteins (e.g., cytokines, chemokines, and inflammatory proteins) The effects on blood biomarkers such as steroids will be determined by protein profiling. This allows for pharmacokinetic studies (e.g., area under the concentration-time curve) to be performed. tion time curve (AUC) and the observed maximum concentration (C max ) can be evaluated by The safety of intravenously administered anti-CD40 antibodies is also assessed by physical examination of the subject; Evidence of reactions, systemic injection-related reactions, and other allergic reactions; electrocardiogram; laboratory tests; vital signs and other adverse events such as infusion-related reactions (IRRS). You can also filter.

[0064] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatment of advanced solid tumors is based on the use of anti-CD4 antibodies in samples obtained from subjects. The amount of antibody against anti-CD40 antibody is determined by measuring the amount of antibody against anti-CD40 antibody. The amount of the antibody can be determined by any method known in the art in light of this disclosure, for example, ELISA. It can be measured by

[0065] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatments for advanced solid tumors are based on terminal half-life, target saturation, and concentration-time profiles. Area under the curve (AUC) and maximum observed concentration (C max ) and other pharmacokinetic parameters were evaluated. The serum sample is analyzed to determine the level of erythrocyte proliferation, which is known in the art in light of this disclosure. The concentration of anti-CD40 antibody is then determined by any known method, e.g., non-compartmental analysis. Pharmacokinetic parameters were analyzed by non-compartment analysis (NCA) to determine the pharmacokinetics of State parameters, e.g., AUC, C max , terminal half-life (T 1 / 2 ), after i.v. Total systemic clearance (CL), terminal volume of distribution (V z ), bioavailability (CL / F) and terminal volume of distribution across bioavailability Product(V z Calculate the formula:

[0066] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatment for advanced solid tumors is targeted therapy with rapid decline in serum concentrations. For example, the half-life of an anti-CD40 antibody is 100 mg / dose. The total dose of anti-CD40 antibody administered per kg of subject is approximately 600 μg / kg. kg, the time is about 10 to 16 hours, preferably about 13 hours, or The total dose of anti-CD40 antibody administered per dose is 1200 mg / kg of subject's body weight. In the case of a dose of 1 μg / kg or more, the time is about 20 to 28 hours, preferably 24 hours.

[0067] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatment for advanced solid tumors requires a dose of 1000 mg / kg of subject's body weight. μg / kg~1400μg / kg, 1050μg / kg~1350μg / kg, 1100 μg / kg~1300μg / kg, 1150μg / kg~1250μg / kg, or 11 Achieve target saturation at concentrations between 50 μg / kg and 1200 μg / kg. For example, saturation concentrations is 1000μg / kg, 1050μg / kg, 1100μg / kg, 1150μg / k g, 1200μg / kg, 1250μg / kg, 1300μg / kg, 1350μg / k g, or 1400 μg / kg, or any dose therebetween.

[0068] According to an embodiment of the present invention, the average C max and AUC 0~24h The increase is 1200μ at doses less than 1200 μg / kg, it was more than dose-proportional, and at doses 1200 μg / kg and above, it was less than dose-proportional. In some embodiments, the clinically proven safe administration and / or administration of anti-CD40 antibodies or clinically proven and safe treatment of advanced solid tumors by assessing CD40 receptor occupancy. For example, the number of B cells and T cells in the peripheral blood after injection of an anti-CD40 antibody is measured. The percentage of cytotoxic and NK cells is measured normalized to pre-infusion levels. According to embodiments of the present invention, the doses of B cells, T cells, and natural killer (NK) cells Independent margination is achieved after injection of anti-CD40 antibodies, whereas dose-dependent B cell margination is achieved after injection of anti-CD40 antibodies. Restoration of N is achieved in agreement with observations made with competing anti-CD40 agonist antibodies. K cells and T cells were reduced in number in the peripheral blood after infusion, and the T cells and natural The normal killer (NK) cells are subsequently fully recovered.

[0069] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatments for advanced solid tumors include MCP-1, IP-10, MIP- 1β, MIP-1α, IFN-γ, TNF-α, IL12p70, IL-2, IL-6, Serum cytokines and chemokines, including but not limited to IL-8 and IL-12 These data are based on the results of anti-CD40 antibody treatment. B cells, T cells to demonstrate changes in PD in cell numbers and / or activation status after infusion This complements flow cytometry testing of the NK compartment, bone marrow, and NK compartment. For example, after injection of anti-CD40 antibody, peripheral chemokines (MCP-1, IP-10, and MIP- 1β) is prominent in the peripheral blood and peaks 1 to 4 hours after injection. Consistent with cell activation, cytokines (IFN-γ, TNF-α, and IL12p70) and Mokines (MIP-1α and IL-8) were also observed, although to a lesser extent, and other C Increased IL-6 levels associated with cytokine storm induction by D40 agonist antibody There are not many after injection.

[0070] In some embodiments, the clinically proven safe administration of anti-CD40 antibodies and / or Clinically proven and safe treatment for advanced solid tumors involves targeting HLA-DR, CD54, and CD80 and flow cytometry of cell surface activation markers such as CD86. Licensing of APCs / DCs in blood samples using any suitable methodology, including but not limited to: This is assessed by measuring the activation of B and T cell subsets in combination with the IL-1 receptor. According to an embodiment of the present invention, these activation markers are expressed by the anti-CD40 antibody. CD40 is increased in activated B cells and monocytes after antibody infusion.

[0071] Abbreviation: β-hCG β-human ciliary gonadotropin ADA anti-drug antibodies ADCC antibody-dependent cell-mediated cytotoxicity AE (treatment-emergent) adverse events ALT alanine transaminase Anti-HCV anti-hepatitis C antibody APC antigen presenting cells AST aspartate transaminase AUC Area under the serum concentration versus time curve BLRM Bayesian logistic regression model CI confidence interval C max Maximum observed serum concentration CR complete response CRF(s) Case Record Form(s) (paper or electronic format as appropriate for this study) CRS cytokine release syndrome CT Computed Tomography CTCAE Common Terminology Criteria for Adverse Events DC dendritic cells DLT dose-limiting toxicity DOR duration of response ECG electrocardiogram ECOG Eastern Cooperative Oncology Group eDC Electronic Data Capture EWOC Escalation with Overdose Control FcγR Fcγ receptor FSH follicle-stimulating hormone GCP: Standards for conducting clinical trials of pharmaceuticals Good Laboratory Practice (GLP) HBsAg Hepatitis B surface antigen hCD40tg human CD40-transgenic HIV human immunodeficiency virus ICF Informed Consent Form ICH International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use IEC Independent Ethics Committee IRB Institutional Review Board irCR immune-related complete response irRC immune-related response criteria IRRs Infusion-related reactions (IRRS) IT intratumoral IV (intravenous) LLOQ Lower limit of quantification MAD maximum dose mCRM revised continuous revaluation method MRI Magnetic Resonance Imaging MTD maximum capacity NCI National Cancer Institute NK Natural Killer NSCLC non-small cell lung cancer ORR Objective Response Rate PCWG Prostate Cancer Clinical Trials Working Group PD Pharmacodynamics PFS Progression-free survival PK Pharmacokinetics Verification of POM mechanism of action PQC Product Quality Complaint PR partial response RBC red blood cells RECIST Response Evaluation Criteria in Solid Tumors RP2D Phase II recommended dose SC subcutaneous SET Safety Assessment Team SUSAR Suspected unexpected serious adverse reaction TEAE Treatment-emergent adverse events T max Time to maximum observed serum concentration TRAF Tumor necrosis factor receptor activator Vd distribution volume WBC white blood cell [Example]

[0072] Example 1: Safety, Pharmacokinetics, and Drug Delivery of Antibody A in Patients with Advanced Solid Tumors A Phase I Open-Label Study of Mechanics Antibody A is an agonistic human monoclonal (IgG1) antibody that targets CD40. This Phase I, open-label study is investigating the treatment of advanced solid tumors. Safety, Pharmacokinetics, and Clinical Trials of Antibody A Administered as an IV Infusion in Patients with Solid Tumors and pharmacodynamics were evaluated, and the recommended Phase 2 dose (RP2D) and schedule were determined. The second part of the study is designed to generate additional safety data. The therapeutic efficacy of Antibody A will be demonstrated in patients who have failed or are unable to tolerate approved effective therapies. Expanded cohort of subjects with unclassified non-small cell lung cancer (NSCLC), pancreatic cancer, and cutaneous melanoma This will be investigated in the court.

[0073] Study design overview Part 1, Dose Escalation: Escalating doses of Antibody A starting at 75 μg / kg are recommended for Phase II. Amended serial relapse in subjects with advanced solid tumors to determine the recommended dose (RP2D) The following factors were investigated in the design of the modified continual reassessment method (mCRM): Doses are increased by no more than half-logarithmic (3.2-fold) dose increments. Maximum tolerated dose of antibody A Until the maximum-tolerated dose (MTD) and / or RP2D are defined, or until Dose escalation will continue until the maximum-administered dose (MAD) is reached. do.

[0074] MTD will use the EWOC principle during the DLT evaluation period to develop a statistical model (BLRM). The highest antibody A efficacy was revealed from the evaluation of the derived PK / PD and safety data. The dosage. · The MAD is defined as the maximum dose of antibody A administered. RP2D is a Bayesian Logistic Regression model. All available PK / PD, safety, and efficacy data were collected according to the BLRM. This will be determined after a review of the data.

[0075] Part 2, Dose Expansion: Antibody A was evaluated for safety and PK / pharmacodynamics, respectively. , PD) characteristics and have NSCLC, pancreatic cancer, and cutaneous melanoma To evaluate the efficacy of this drug in subjects, an expansion cohort of approximately 30 subjects will be conducted. It will be administered at RP2D.

[0076] Part 2 (dose expansion) will begin after the RP2D is determined and will include: (1) evaluation of the efficacy and safety of the drug in selected disease populations; (2) collect additional information on the safety and PK / PD characteristics of Antibody A, which is being investigated for NSCLC and pancreatic cancer; The clinical activity of the test drug will be evaluated in subjects with cancer and cutaneous melanoma. Cohorts consist of approximately 30 subjects each.

[0077] Biomarker Substudy: Evaluating additional biomarkers to assess innate immunity in tumors Define the effect of Antibody A on the immune response and adaptive immune response.

[0078] Figure 1 shows a diagram of the study design.

[0079] subject Subjects must be 18 years of age or older and must be enrolled in a clinical trial with the Eastern Cooperative Oncology Group (ECG) or the US National Cancer Institute (CIRC). The European Cooperative Oncology Group (ECOG) performance status score must be 0 or 1. .

[0080] Part 1: Advanced or refractory solid malignant tumors of any type that are metastatic or unresectable Subjects with are eligible for enrollment in Part 1. Subjects must have received all standard treatment options. or are no longer eligible for additional standard treatment options.

[0081] Part 2: Patients with histologically or cytologically confirmed NSCLC, pancreatic cancer, or cutaneous melanoma Subjects who: are eligible for enrollment in Part 2. Subject cohorts include, for example:

[0082] Cohort 2A: Histologically or cytologically confirmed NSCLC Stage IV disease Prior treatment with at least two approved systemic therapies, one of which was white blood cell Must be a gold-containing regimen At least one measurable tumor lesion by RECIST v1.1

[0083] Cohort 2B: Histologically or cytologically confirmed adenocarcinoma of the pancreas Unresectable, locally advanced (Stage III), or metastatic (Stage IV) disease Prior treatment with at least one approved systemic therapy At least one measurable tumor lesion by RECIST v1.1

[0084] Cohort 2C: Histologically or cytologically confirmed cutaneous melanoma Unresectable (stage III) or metastatic (stage IV) disease Prior treatment with at least one approved systemic therapy At least one measurable tumor lesion by RECIST v1.1

[0085] The second part of the trial is underway.

[0086] Test drug Antibody A is supplied as a lyophilized cake or as a frozen liquid. The formulation is 20 mg / Contains 40 mg / mL or 40 mg / mL antibody A.

[0087] Dosage and Administration Doses will be titrated from a starting dose of 75 μg / kg. Administration will be by IV infusion. The initial schedule of administration is every 14 days (Q14d) in a 28-day cycle. Administration will initially be by IV infusion over 2 hours. .

[0088] Recommended supportive care before and after infusion Prior to each infusion of Antibody A, subjects will receive pre-infusion medication as described in Table 1. do.

[0089] [Table 1] IV = intravenous a Explicitly mandated by the Safety Evaluation Team (SET) If not, any

[0090] Study design and cohorts with pre-infusion of corticosteroids and corticosteroids The study design and cohorts without pre-infusion are shown in Figure 2 .

[0091] After each infusion of Antibody A, subjects who experienced Antibody A-related toxicity during or after the previous infusion were included in Table 2 Post-infusion medications should be taken as described in.

[0092] [Table 2] IV = intravenous a In the absence of symptoms of JNJ-64457107-related toxicity, post-infusion medications will be discontinued until the end of the infusion. may be given up to 48 hours after administration. b The investigator will use clinical judgment if further corticosteroid support is needed. It is necessary to use c Oral or IV administration will begin 12 hours after the first dose of study drug. It is recommended that administration be limited to 48 hours after administration.

[0093] evaluation Safety evaluation Safety evaluation was based on medical review of adverse event reports, clinical laboratory test results, and cardiac evaluation at each time point. Electrocardiogram (ECG), vital sign measurements, physical examination, ECOG performance score, and Based on other safety assessments.

[0094] Pharmacokinetics and immunogenicity Blood or serum samples were used to analyze PK and antibody responses in all subjects participating in the study. The immunogenicity of antibody A was evaluated by measuring the serum concentration of antibody A (approximately 5 mL) and the anti-antibody concentration. A antibody (approximately 7.5 mL for the combined PK and immunogenic samples, otherwise Venous blood will be collected for evaluation of immunogenicity (5 mL for the sample alone). Samples were collected, and if both PK and immunogenicity samples were collected, for each time point, each serum Divide the sample into three equal aliquots (one each for PK, anti-antibody A, and backup). otherwise, if only PK samples were collected, two equal aliquots were used for each time point. It is divided into

[0095] Serum samples are analyzed by or under the supervision of the sponsor to identify validated specific and The concentration of Antibody A will be determined using a sensitive assay. MescoScale Discovery (MSD) Pl, verified under client supervision A formal assay will be used to detect and characterize antibodies against Antibody A. All samples collected to detect antibodies to antibody A were analyzed for serum antibody A concentrations. Evaluating these factors will allow interpretation of antibody data.

[0096] Biomarkers To investigate the molecular mode of action of Antibody A and to predict response to therapy Biomarkers are evaluated to identify potential biomarkers that may be useful in treating this condition. Biomarker objectives for the study were CD40 receptor occupancy, innate immune response, CD4 Examination of activation markers, Fc-dependent effector functions, and CD40 expression in tumor tissues Optional biomarker sub-tests include: Evaluate immune cell changes following CD40 engagement to identify combination opportunities To do this, pre-drug and post-drug biopsies will be used.

[0097] Efficacy evaluation Efficacy assessment was performed by computed tomography (CT) or magnetic resonance imaging for all subjects. This will include MRI and bone scans for subjects with prostate cancer. The answer is according to Response Evaluation in Solid Tumors (RECIST) v1.1 criteria and immune-related The Prostate Cancer Clinical Trials Working Group will evaluate patients according to the IRRC. The PCWG3 criteria were used to assess disease response in subjects with prostate cancer. The relationship between PK, including receptor occupancy, and PD will be investigated and reported separately. This will be reported in.

[0098] statistical methods Data will be summarized using descriptive statistics. Continuous variables will be summarized by number of observations, as appropriate. Data will be summarized using means, standard deviations, medians, and ranges. Categorical values ​​will be used where appropriate. The distribution of time-to-event endpoints is summarized using the number of observations and percentages. It is estimated using the Plan-Meier method.

[0099] result Demographics and Breakdown: 95 patients, ages 18-80 (median 59), received corticosteroids. Seven cohorts (n=50, 75μg / kg to 2000μg / kg) with and without steroids Five cohorts (n=45, 75μg / kg to 1200μg / kg) without steroids were enrolled. Patients were registered and received 1 to 26 (median 3) cycles of antibody A (Figure 2). Most patients progressed. Discontinuation due to sexual illness (n=62, 76%)

[0100] Safety Results As shown in Table 3, the majority of adverse events (AEs) were grade 1 (G1) or The incidence of AEs was 2 (G2), and the number of patients with grade 3 (G3) or higher AEs was limited.

[0101] [Table 3]

[0102] As shown in Figure 3, infusion-related reactions (IRRs) were reported in 51% of patients (G1-G2). G2: 50%, G3: 1%). The most common IRRs (>10%) were pruritus (31%) and rash. (15%), cold sensation (13%), and flushing (12%). Based on the high incidence of pruritus, A new pre-medication regimen was implemented. Cetirizine and montelukast were administered 3 days before each antibody A infusion. administered as a pre-medication / post-medication up to 3 days after the initial dose and significantly reduced the IRR. and no pruritus was reported.

[0103] Two dose-limiting toxicities (DLTs) were observed: 1200 μg / 1200 μg / kg for 5 days of continuous G3 headache and no corticosteroids / kg, G3 ALT / AST increase + G2 bilirubin increase was reported.

[0104] Pharmacokinetic results Preliminary PK of Antibody A demonstrated target-mediated drug disposition with rapid decline in serum concentrations. The half-life was approximately 13 hours at 600 μg / kg, and g / kg for approximately 24 hours (Figure 4). No accumulation was observed after multiple biweekly administrations. Based on the data collected, target saturation was observed at approximately 1200 μg / kg. max Reach and AUC 0~24hThe increase in α (Figure 5) was greater than dose-proportional at doses less than 1200 μg / kg. The immunogenicity data showed that antibody A was showed a low incidence of antibodies (approximately 10.5%).

[0105] Pharmacodynamic results Dose-dependent margination of B cells, T cells, and natural killer (NK) cells was observed with antibodies. A dose-dependent B cell recovery was observed after injection of a competitive anti-CD40 agonist antibody. NK cells and T cells were significantly increased at all doses tested except for the lowest dose (75 μg / kg). Both levels were reduced in peripheral blood after infusion at all doses, and both levels were fully recovered by day 8 of the study. See Figures 6A-6C.

[0106] Peripheral chemokines (MCP-1, IP-10, and MIP-1β) are expressed in peripheral blood. This is significant and peaks 1-4 hours after injection, consistent with myeloid cell activation. Tokine (IFN-γ, TNF-α and IL12p70) and chemokine (MIP-1α and IL-8) were also observed, albeit to a lesser extent. The levels of IL-6, which is associated with the induction of cytokine storm by antibody A, increased after injection of antibody A. , but not many. See Figures 7A to 7I.

[0107] Phenotypic staining of peripheral B cells indicates activation / maturation consistent with data from other agonist antibodies The increased fluorescence intensity of markers (HLA-DR, CD54, CD80, and CD86) (Fig. 8A to Fig. 8D).

[0108] Clinical Activity Results Early signs of clinical activity were seen in patients with renal cell carcinoma and those with long-term stable disease for 6 months or more. The results included partial responses in 10 patients.

[0109] Consideration CD40 agonist antibody A has a manageable safety profile with favorable PK and PD properties. Preliminary PK of Antibody A was observed to be above 1200 μg / kg with moderate variability. Antibody A inhibits selected chemokines, particularly MCP- Increased levels of IP-10 and IP-10, as well as peripheral blood pressure, B cells, T cells, and NK cells after infusion The remaining peripheral B cells showed increased activation / maturation markers. It showed large.

[0110] Those skilled in the art may make changes to the above-described embodiments without departing from the broad inventive concept. It will be understood, therefore, that the invention is not limited to the particular embodiments disclosed. It does not cover modifications within the spirit and scope of the invention as defined by the specific description. It is understood that it is intended to

Claims

1. Clinically proven safety of anti-CD40 antibodies in human subjects diagnosed with advanced solid tumors a pharmaceutical composition comprising the antibody and a pharmaceutically acceptable carrier, and administering to the subject intravenously an antibody comprising a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises the heavy chain complementarity determining regions of SEQ ID NOs: 1, 2, and 3, respectively ( HCDR) the amino acid sequences of HCDR1, HCDR2, and HCDR3, The variable regions are the light chain complementarity determining regions (LCDRs) of SEQ ID NOs: 4, 5, and 6, respectively. the total dose of the antibody administered, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3; is 50 μg / kg to 2500 μg / kg per kg of the subject's body weight per administration. g, preferably 75 μg to 2000 μg / kg.

2. The antibody comprises a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO: 7 and a nucleotide sequence of SEQ ID NO:

8. The method of claim 1, comprising a light chain variable region (VL) having the amino acid sequence:

3. The antibody comprises a heavy chain (HC) having the amino acid sequence of SEQ ID NO: 9 and an amino acid sequence of SEQ ID NO:

10. The method of claim 1 or 2, comprising a light chain (LC) having the sequence:

4. The total dose of the anti-CD40 antibody administered per administration is 75μg / kg, 200μg / kg, 400μg / kg, 600μg / kg, 70 0μg / kg, 800μg / kg, 900μg / kg, 1000μg / kg, 1100μ g / kg, 1200μg / kg, 1300μg / kg, 1400μg / kg, 1500μ 1800 μg / kg, 1800 μg / kg, or 2000 μg / kg, or any value therebetween. The method according to any one of claims 1 to 3, wherein the dosage is any desired amount.

5. The total dose of the pharmaceutical composition is administered intravenously to the human subject over a period of about two hours, preferably Preferably, the pharmaceutical composition is administered intravenously to the human subject repeatedly, more preferably once every two weeks. The method of any one of claims 1 to 4, wherein the administration is intravenous.

6. further administering a therapeutic agent to said human subject before or after said administration of said anti-CD40 antibody. Preferably, the therapeutic agent is a corticosteroid, an antihistamine, an antipyretic, a H ２ - antagonists, and antiemetics.

1. The method according to claim 1.

7. in combination with the anti-CD40 antibody, an effective amount of cetirizine and montelukast administering at least one of the compounds to the subject reduces infusion-related reactions (IRRS) or pruritus. and preferably cetirizine and montelukast further comprise reducing the reaction of The method according to any one of claims 1 to 5, wherein the anti-CD40 antibody is administered within 3 days before and within 3 days after the administration of the anti-CD40 antibody.

7. The method according to any one of claims 6 to 6.

8. The total dose of the anti-CD40 antibody administered per administration is When the anti-CD40 antibody is administered at a concentration of 600 μg / kg or less, the anti-CD40 antibody is administered for approximately 10 to 16 hours, preferably The anti-CD40 antibody has an in vivo half-life of about 13 hours, or When the total body dose is 1200 μg / kg or more per kg of the subject's body weight, The anti-CD40 antibody has an in vivo half-life of about 20 to 28 hours, preferably 24 hours. The method according to any one of claims 1 to 7.

9. When the pharmaceutical composition is administered intravenously to the human subject repeatedly, the human subject The method according to any one of claims 1 to 8, wherein the anti-CD40 antibody is not accumulated.

10. The total dose of the anti-CD40 antibody administered per administration is about 1000-1400 μg / kg, preferably 1200 μg / kg of body weight of said subject 1 to 3, wherein the administration of the anti-CD40 results in target saturation when the anti-CD40 is at a dose of 1 μg / kg.

10. The method according to any one of claims 9 to 10.

11. The administration of the anti-CD40 antibody results in an increase in B cells, T cells, and / or T cells in the peripheral blood of the human subject. and margination of one or more cells selected from the group consisting of natural killer (NK) cells. and subsequent recovery of said cells. 。

12. The administration of the anti-CD40 antibody results in an increase in MCP-1, IP one or more selected from the group consisting of MIP-10, MIP-1β, MIP-1α, and IL-8 The method according to any one of claims 1 to 11, wherein an increase in chemokines of

13. The administration of the anti-CD40 antibody results in increased levels of IFN-γ, TNFR, and TNFR in the peripheral blood of the human subject. and increasing one or more cytokines selected from the group consisting of F-α, and IL12p70. The method according to any one of claims 1 to 12,

14. The administration of the anti-CD40 antibody may result in the activation of one or more activation markers on peripheral blood B lymphocytes. The activation markers are HLA-DR, CD54, CD80, and CD 14. The method of any one of claims 1 to 13, wherein the compound is selected from the group consisting of:

15. The human subject has at least a partial response or has long-term disease for six months or more. The method of any one of claims 1 to 14, wherein the method is stable to

16. Clinically proven safety of anti-CD40 antibodies in human subjects diagnosed with advanced solid tumors a pharmaceutical composition comprising the antibody and a pharmaceutically acceptable carrier, and administering to the subject intravenously an antibody comprising a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises the heavy chain complementarity determining regions of SEQ ID NOs: 1, 2, and 3, respectively ( HCDR) the amino acid sequences of HCDR1, HCDR2, and HCDR3, The variable regions are the light chain complementarity determining regions (LCDRs) of SEQ ID NOs: 4, 5, and 6, respectively. the total dose of the antibody administered, comprising the amino acid sequences of LCDR1, LCDR2, and LCDR3; is 600 μg / kg, 700 μg / kg, per kg of the subject's body weight per administration. g, 800 μg / kg, 900 μg / kg, 1000 μg / kg, 1100 μg / kg, about 600 μg / kg to about 1200 μg / kg, such as 1200 μg / kg, preferably The human subject has been diagnosed with non-small cell lung cancer (NSCLC), pancreatic cancer, or cutaneous melanoma. There is a way.

17. The antibody comprises a heavy chain variable region (VH) having the amino acid sequence of SEQ ID NO: 7 and a nucleotide sequence of SEQ ID NO:

8. The method of claim 16, comprising a light chain variable region (VL) having the amino acid sequence:

18. The antibody comprises a heavy chain (HC) having the amino acid sequence of SEQ ID NO: 9 and an amino acid sequence of SEQ ID NO:

10. The method of claim 16 or 17, comprising a light chain (LC) having the sequence:

19. 1. A pharmaceutical composition comprising an anti-CD40 antibody and a pharmaceutically acceptable carrier, wherein the antibody: The heavy chain variable region comprises a heavy chain variable region and a light chain variable region, and the heavy chain variable region is represented by SEQ ID NOs: 1, 2, and 3, respectively. and three heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. the light chain variable region comprises the light chain complementarity determinations of SEQ ID NOs: 4, 5, and 6, respectively. The LCDR comprises the amino acid sequences of LCDR1, LCDR2, and LCDR3. The total dose of the antibody administered is 50 μg / kg of subject body weight per administration. 75 μg / kg to 2500 μg / kg, preferably 75 μg / kg to 2000 μg / kg, Pharmaceutical composition.

20. An anti-CD40 antibody for use in medicine, said antibody comprising a heavy chain variable region and a light chain variable region. The heavy chain variable region comprises a heavy chain complementary region of SEQ ID NOs: 1, 2, and 3, respectively. The amino acid sequences of the HCDR1, HCDR2, and HCDR3 are included in the The light chain variable regions are composed of light chain complementarity determining regions (LCDRs) of SEQ ID NOS: 4, 5, and 6, respectively. the total amino acid sequence of the antibody to be administered, comprising LCDR1, LCDR2 and LCDR3; The dosage is 50 μg / kg to 2500 μg / kg of the subject's body weight per administration. kg, preferably 75 μg / kg to 2000 μg / kg.

21. An anti-CD40 antibody for use in the treatment of advanced solid tumors, said antibody comprising The heavy chain variable region is a sequence represented by SEQ ID NO: 1, 2, and 3, and the light chain variable region is a sequence represented by SEQ ID NO: 3, 4, and 5, respectively. and amino acids of the heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, and HCDR3. the light chain variable region comprises the light chain complementarity determining sequences of SEQ ID NOs: 4, 5, and 6, respectively; The LCDR region (LCDR) comprises the amino acid sequences of LCDR1, LCDR2, and LCDR3, and is administered The total dose of the antibody administered per administration is 50 μg / kg of subject body weight. anti-C, which is 75 μg / kg to 2500 μg / kg, preferably 75 μg / kg to 2000 μg / kg; D40 antibody.

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