Use of antibody-drug conjugates in the manufacture of drugs for the prevention and / or treatment of cancer

Abstract

The present invention relates to the use of antibody-drug conjugates, or pharmaceutically acceptable salts, stereoisomers, metabolites, or solvent compounds thereof, in the manufacture of agents for the prevention and / or treatment of cancer. Specifically, the present invention provides the use of anti-B7-H4 antibody-drug conjugates, or pharmaceutically acceptable salts, stereoisomers, metabolites, or solvent compounds thereof, in the manufacture of agents for the prevention and / or treatment of cancer, particularly cholangiocarcinoma with low B7-H4 expression. The present invention also provides drug formulations comprising B7-H4 antibody-drug conjugates, or pharmaceutically acceptable salts, stereoisomers, metabolites, or solvent compounds thereof.

Description

Technical Field

[0001] The present application belongs to the medical field and relates to the use of antibody-drug conjugates in the manufacture of drugs for the prevention and / or treatment of cancer diseases.

Background Art

[0002] Cholangiocarcinoma originates from cholangiocytes and accounts for about 3% of gastrointestinal tumors and 10 - 15% of malignant tumors of the hepatobiliary system. The incidence rate of cholangiocarcinoma is higher in Asian countries than in Western countries, with an incidence rate of 113 per 100,000 in men and 50 per 100,000 in women. The only treatment for cholangiocarcinoma is radical surgery. However, this disease does not exhibit characteristic clinical symptoms at the early stage, and the majority of patients are in the terminal state at the time of diagnosis, so the prognosis is extremely poor, and the 5-year survival rate is about 10%. In addition, up to 50% of cholangiocarcinoma recurs within 1 year after surgical resection. In unresectable locally advanced and metastatic bile duct tumors (including intrahepatic and extrahepatic cholangiocarcinoma, gallbladder cancer, and papillary cancer), in the standard first-choice chemotherapy regimen (gemcitabine + cisplatin), the median survival period is only 11.7 months, and the prognosis is extremely poor. Drugs targeting BRAF, IDH1, FGFR2, HER2, and PD-1 / PD-L1 have been approved as therapeutic drugs for advanced biliary malignancies, but their therapeutic effects are limited, and there is an urgent need for new treatment methods to improve the treatment limitations of patients with advanced cholangiocarcinoma.

[0003] Antibody-drug conjugates (ADCs) are a type of drug that chemically conjugates a bioactive cytotoxic agent to a monoclonal antibody that acts as a carrier for transporting the cytotoxic agent. Currently, ADCs are mainly used in the oncology field and generally require high expression of antigen targets in tumor cells. B7-H4 is a newly discovered member of the B7 family that plays a crucial role in various cellular biological processes such as differentiation, proliferation, and apoptosis, and may influence tumor cell invasion and metastasis. The B7 family is also an important co-stimulatory molecule that may influence T cell proliferation, B cell activation, and other processes. Studies have shown that B7-H4 expression is high in many tumors, including cholangiocarcinoma, breast cancer, endometrial cancer, non-small cell lung cancer, ovarian cancer, gastric cancer, and pancreatic cancer, while its expression is limited in normal tissues, making B7-H4 a promising target for ADCs.

[0004] B7-H4 is highly expressed in cholangiocarcinoma, and patients with high B7-H4 expression have a poor prognosis, suggesting that B7-H4 could be a potential target in the treatment of cholangiocarcinoma. While it has been reported that antibodies and ADCs targeting B7-H4 can be used in cancers with high B7-H4 expression, including cholangiocarcinoma, the therapeutic effect of these drugs in cholangiocarcinoma with low B7-H4 expression is unknown. [Overview of the Initiative]

[0005] According to a first aspect of the present invention, the use of an anti-B7-H4 antibody-drug conjugate, or a pharmaceutically acceptable salt thereof, stereoisomer, metabolite, or solvent compound thereof, in the manufacture of a drug for the prevention and / or treatment of cancer, wherein the anti-B7-H4 antibody-drug conjugate is of formula (I): [ka] [In the formula, y represents a number between 1 and 20, preferably between 2 and 8, more preferably between 4 and 6 or between 7 and 8; y is a decimal or an integer, even more preferably between 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3 or 6.4; Ab is an anti-B7-H4 antibody or its antigen-binding fragment, comprising a variable region of the antibody heavy chain and a variable region of the antibody light chain, wherein the variable region of the antibody heavy chain comprises HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NOs. 1, 2, and 3, respectively; and the variable region of the antibody light chain comprises LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NOs. 4, 5, and 6, respectively. The invention provides having a structure represented by; the cancer being a cancer with low B7-H4 expression.

[0006] In some embodiments, the anti-B7-H4 antibody or its antigen-binding fragment is a humanized antibody or its fragment.

[0007] In some embodiments, the humanized antibody comprises a heavy chain variable region shown in SEQ ID NO: 7 and a light chain variable region shown in SEQ ID NO: 8; preferably, the humanized antibody comprises a heavy chain shown in SEQ ID NO: 9 and a light chain shown in SEQ ID NO: 10.

[0008] In some embodiments, cancers with low B7-H4 expression are characterized by a B7-H4 expression level of IHC 1+ or 2+.

[0009] In some embodiments, cancers with low B7-H4 expression are cases in which, numerically, B7-H4 expression is IHC 1+ or 2+ in at least 1% of tumor cells in a sample from a clinical patient, or cases in which B7-H4 expression is IHC 1+ or 2+ in numerically, 0-1% of cells.

[0010] In some embodiments, cancers with low B7-H4 expression are cholangiocarcinomas with low B7-H4 expression, preferably cholangiocarcinomas with B7-H4 expression levels of IHC 1+ or IHC 2+.

[0011] In some embodiments, cancers with low B7-H4 expression are cholangiocarcinomas with low B7-H4 expression, preferably cholangiocarcinomas with B7-H4 expression levels of IHC 1+ or IHC 2+ in clinical trials.

[0012] In some embodiments, the prevention and / or treatment comprises administering to a patient a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, at a dose of 0.5 to 15 mg / kg body weight, preferably 3 to 10 mg / kg body weight.

[0013] In some embodiments, the prevention and / or treatment comprises administering to a patient, via an oral, parenteral, or transdermal route, a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, the parenteral route being selected from intravenous, subcutaneous, or intramuscular injection.

[0014] In some embodiments, the prevention and / or treatment comprises administering to a patient a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, wherein the drug is in an injectable form, e.g., subcutaneous or intravenous; preferred injectable forms are injectables or lyophilized powders for injection comprising the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, along with buffers, stabilizers, pH adjusters, and optionally present surfactants.

[0015] A second aspect of the present invention provides a method for preventing and / or treating cancer with low B7-H4 expression, comprising administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I) above, or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, to a subject in need, preferably at 0.5 to 15 mg / kg body weight, more preferably at 3 to 10 mg / kg body weight; or The treatment involves administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I) described above, or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, as a concomitant drug.

[0016] Cancers with low B7-H4 expression are those in which the B7-H4 expression level is IHC 1+ or 2+; preferably, cancers with low B7-H4 expression are cholangiocarcinomas with low B7-H4 expression; more preferably, cancers with low B7-H4 expression are cholangiocarcinomas in which the B7-H4 expression level is IHC 1+ or IHC 2+.

[0017] In some embodiments, cancers with low B7-H4 expression are cases in which, numerically, B7-H4 expression is IHC 1+ or 2+ in at least 1% of tumor cells in a sample from clinical patients, or cases in which B7-H4 expression is IHC 1+ or 2+ in numerically, 0-1% of cells; preferably, cancers with low B7-H4 expression are cholangiocarcinomas with low B7-H4 expression, and more preferably, cancers with low B7-H4 expression are cholangiocarcinomas with IHC 1+ or IHC 2+ B7-H4 expression levels in clinical trials.

[0018] The drugs used in the aforementioned methods of prevention and / or treatment are administered orally, parenterally, or transdermally, with parenterally being selected from intravenous, subcutaneous, or intramuscular injection.

[0019] In some embodiments, the drugs in the above-mentioned prevention and / or treatment include injectable forms, such as subcutaneous or intravenous injection; a more preferred injectable form is an injectable preparation or lyophilized powder for injection comprising an anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, and buffers, stabilizers, pH adjusters, and optionally present surfactants.

[0020] According to a third aspect of the present invention, a drug formulation comprising a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I) above, or a pharmaceutically acceptable salt, metabolite, or solvent compound thereof, further comprising a pharmaceutically acceptable carrier, wherein the anti-B7-H4 antibody-drug conjugate is administered at a dose of 0.5 to 15 mg / kg body weight, preferably 3 to 10 mg / kg body weight, and is used for the treatment of cancer with low B7-H4 expression, wherein the cancer with low B7-H4 expression is a cancer with a B7-H4 expression level of IHC 1+ or 2+, preferably a cholangiocarcinoma with low B7-H4 expression, and more preferably a cholangiocarcinoma with a B7-H4 expression level of IHC 1+ or 2+.

[0021] In some embodiments, cancers with low B7-H4 expression are those in which, in samples from clinical patients, B7-H4 expression is IHC 1+ or 2+ in at least 1% of tumor cells numerically related to IHC, or IHC 1+ or 2+ in 0-1% of cells numerically related to IHC; preferably, cancers with low B7-H4 expression are cholangiocarcinomas with low B7-H4 expression; more preferably, cancers with low B7-H4 expression are cholangiocarcinomas with IHC 1+ or IHC 2+ B7-H4 expression levels in clinical trials.

[0022] The anti-B7-H4 antibody-drug conjugate of the present invention significantly inhibits the proliferation of xenografts of human cholangiocarcinoma LD1-0060-202214 with low B7-H4 expression in nude mice, and does not cause significant weight loss or other abnormal symptoms in the mice, thus demonstrating excellent prospects for clinical use.

Brief Description of Drawings

[0023] [Figure 1] Figure 1 shows the efficacy of an anti-B7-H4 antibody-drug conjugate in subcutaneous transplantation of human cholangiocarcinoma LD1-0060-202214 in mice. [Figure 2] Figure 2 shows the effect of an anti-B7-H4 antibody-drug conjugate on the body weight of mice with human cholangiocarcinoma LD1-0060-202214. Detailed Description of the Invention

[0024] I. Terminology To better understand the present invention, some technical terms and scientific terms are defined in detail below. Unless explicitly defined otherwise herein, all technical terms and scientific terms used in this specification have meanings that can be generally understood by those skilled in the technical field of the present invention.

[0025] The three-letter codes and one-letter codes of amino acids used in this specification are those described in J. Biol. Chem, 243, p3558 (1968).

[0026] As used herein, the term "antibody" refers to an immunoglobulin having a tetrameric structure in which two identical heavy chains and two identical light chains are linked via interchain disulfide bonds. The amino acid composition and sequence of the heavy chain constant region vary between immunoglobulins, resulting in different antigenic properties. Thus, immunoglobulins can be classified into five major classes (also called isotypes), namely IgM, IgD, IgG, IgA, and IgE, corresponding to μ heavy chain, δ heavy chain, γ heavy chain, α heavy chain, and ε heavy chain, respectively. Each immunoglobulin class can be further classified into subclasses based on differences in the amino acid composition of the hinge region and the number and position of the disulfide bonds in the heavy chain. For example, IgG can be classified into IgG1, IgG2, IgG3, and IgG4. Light chains are classified into κ (kappa) or λ (lambda) based on the constant region. The five immunoglobulin classes each contain either κ or λ light chains.

[0027] In the present invention, the variable region of the light chain may further include a constant region. The constant region of the light chain includes a human or mouse κ-chain or λ-chain, or a variant thereof.

[0028] Similarly, the variable region of the heavy chain may further include a constant region. The constant region of the heavy chain may include human or mouse IgG1, IgG2, IgG3, or IgG4, or their variants.

[0029] The N-terminal regions (the first approximately 110 amino acids) of both the heavy and light chains exhibit considerable sequence variation and are referred to as the variable region (V region). The remaining C-terminal region has a relatively conserved amino acid sequence and is known as the constant region (C region). Each variable region consists of three hypervariable regions (HVRs) and four relatively conserved framework regions (FRs). The three hypervariable regions determine the antigen-binding specificity of the antibody and are also referred to as complementarity-determining regions (CDRs). The variable regions of the light chain (VL) and heavy chain (VH) each consist of three CDRs and four FRs, arranged sequentially from the N-terminus to the C-terminus as FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The three CDRs of the light chain are referred to as LCDR1, LCDR2, and LCDR3, respectively, and the three CDRs of the heavy chain are referred to as HCDR1, HCDR2, and HCDR3, respectively. The CDR amino acid residues in the VL and VH regions of the antibody or antigen-binding fragment of the present invention conform to the known Kabat numbering rules and the definition of Kabat or AbM in terms of their number and position.

[0030] The term "antigen-presenting cell" or "APC" refers to a cell that presents a foreign antigen on its surface in coexistence with major histocompatibility complex (MHC) molecules. T cells recognize these complexes via the T cell receptor (TCR). Examples of APCs include, but are not limited to, dendritic cells (DCs), peripheral blood mononuclear cells (PBMCs), monocytes, B lymphoblasts, and monocyte-derived dendritic cells (DCs). The term "antigen presentation" refers to the process by which APCs capture an antigen and present it to T cells for recognition, for example, as part of an MHC class I or class II complex.

[0031] The term "B7-H4" refers to a type I transmembrane protein containing four immunoglobulin-like extracellular domains, also known as CD276, which is a member of the human B7 protein family. B7-H4 is an immune checkpoint protein expressed on the surface of antigen-presenting cells or cancer cells and is known to inhibit T cell activation. The term "B7-H4" encompasses all variants or isotypes of B7-H4 that are spontaneously expressed in cells. The antibodies of the present invention may cross-react with B7-H4 from non-human species. Alternatively, the antibodies may be specific to human B7-H4 and not cross-react with B7-H4 from other species. B7-H4, or its variants or isotypes, may be isolated from cells or tissues that spontaneously express it, or may be recombinantly produced using techniques known in the art and techniques described herein. Preferably, the anti-B7-H4 antibody targets human B7-H4 having a normal glycan pattern.

[0032] The term "humanized antibody," also known as a CDR-transplanted antibody, refers to an antibody in which a mouse antibody-derived CDR is transplanted into the framework region of the human antibody variable domain. This overcomes the potent immune response induced by the large mouse protein elements contained in chimeric antibodies. To minimize the decrease in activity associated with reduced immunogenicity, minimal reversion mutations may be introduced into the human antibody variable region to maintain binding activity.

[0033] The term "antigen-binding fragment" refers to an antigen-binding fragment of an antibody or antibody analog, which typically includes at least some of the antigen-binding or variable regions (e.g., one or more CDRs) of the parent antibody. The antibody fragment retains at least some of the binding specificity of the parent antibody. Generally, when binding activity is measured in molar concentration, the antibody fragment retains at least 10% of the binding activity of the parent antibody. Preferably, the antibody fragment retains at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the binding affinity of the parent antibody to the target antigen. Examples of antigen-binding fragments include, but are not limited to, Fab, Fab', F(ab')2, Fv fragments, linear antibodies, single-chain antibodies, nanobodies, domain antibodies, and multispecific antibodies. Modified antibody variants are summarized in Holliger and Hudson (2005) Nat. Biotechnol. 23:1126-1136.

[0034] The term “combination drug” refers to any product containing one or more active ingredients (e.g., antibodies, antibody-drug conjugates, small molecule drugs) in any specified amount, and any product obtained by directly or indirectly combining one or more active ingredients in any specified amount. These drugs may optionally contain appropriate pharmaceutical adjuvants, such as known pharmaceutical carriers and pharmaceutical excipients, including buffers, in the art. Different active ingredients in a combination drug may be administered independently, simultaneously or at different time points, in separate dosage forms to achieve a synergistic effect. In this disclosure, “combination drug” and “formulation” are not mutually exclusive.

[0035] The term "pharmaceutically acceptable salt" refers to a salt of an antibody-drug conjugate that is safe and effective when administered to mammals and retains appropriate physiological activity. The antibody-drug conjugates of the present invention contain at least one amino acid and are therefore capable of forming salts with acids. Non-limiting examples of pharmaceutically acceptable salts include: hydrochloride, hydrobromide, hydroiodide, sulfate, disulfate, citrate, acetate, succinate, ascorbate, oxalate, nitrate, sorbate, hydrogen phosphate, dihydrogen phosphate, salicylate, disodium citrate, tartrate, maleate, fumarate, formate, benzoate, mesylate, esylate, benzenesulfonate, and tosylate. "pharmaceutically acceptable salt" and "pharmaceutically acceptable salt" can be used interchangeably.

[0036] "Solvent compound" refers to a compound of the pharmaceutical solvent compound of the antibody-drug conjugate in the present invention and one or more solvent molecules. Non-limiting examples of solvent molecules include: water, ethanol, acetonitrile, isopropanol, and ethyl acetate.

[0037] The "drug-to-antibody ratio" (DAR) is expressed as y, which is the average amount of cytotoxic drug per antibody in formula (I). The DAR ranges from 1 to 20 cytotoxic drug molecules (D) per antibody molecule. An antibody-drug conjugate represented by general formula (A) consists of a group of antibodies to which a certain number (1 to 20) cytotoxic drug molecules are bound. The DAR of an antibody-drug conjugate obtained by a binding reaction can be measured by conventional analytical techniques such as mass spectrometry, high-performance liquid chromatography (HPLC), and enzyme-linked immunosorbent assay (ELISA). These analytical methods allow for the quantitative distribution of the y value of the antibody-drug conjugate.

[0038] The term "low B7-H4 expression" generally refers to a level of B7-H4 expression in at least 1% of tumor cells in a clinical sample that is determined to be IHC 1+ or 2+ by immunohistochemistry (IHC) testing. Even if 0-1% of tumor cells show IHC 1+ or 2+ B7-H4 expression, this invention is still considered to fall within the range of "low B7-H4 expression." The terms "high B7-H4 expression" and "B7-H4 overexpression" are used interchangeably and generally refer to IHC 3+ expression as determined in clinical trials. [Examples]

[0039] The following embodiments are provided to further illustrate the present invention. However, these embodiments do not limit the scope of the present invention in any way. Unless otherwise specified, the experimental methods described in the embodiments of the present invention were carried out under standard conditions such as those described in the Cold Spring Harbor Laboratory Antibody Technology Experiment Manual and Molecular Cloning Manual, or according to instructions provided by the manufacturers of the raw materials or reagents. Reagents that are not of specific origin are commercially available standard reagents.

[0040] II. Biological Evaluation Methods Example 1. In vivo pharmacodynamics of anti-B7-H4 antibody-drug conjugates in a xenograft model of human cholangiocarcinoma LD1-0060-202214 in NU / NU mice. 1. Experimental Objective We evaluated the in vivo pharmacodynamics of anti-B7-H4 antibody-drug conjugates in a xenograft model of human cholangiocarcinoma LD1-0060-202214 in NU / NU mice.

[0041] 2. Experimental materials 2.1 Information on the Human Cholangiocarcinoma LD1-0060-202214PDX Model LD1-0060-202214, a tumor tissue from human cholangiocarcinoma, was passaged up to FP3+3 generations for this pharmacodynamic study and provided by Shanghai LIDE Biotech Co., Ltd. [Table 1]

[0042] 2.2 Test reagent 1) Anti-B7-H4 antibody (hu2F7 unbound antibody, Shanghai Hansoh BioMedical Co., Ltd., batch number: DS202207): The sequence of the anti-B7-H4 antibody (hu2F7 unbound antibody) is disclosed in WO2019154315A1 as follows.

[0043] The antibody contains the following CDR sequence: [Table 2]

[0044] hu2F7 HCVR EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVAYVSSGGGSTYYSDSVKGRFTISRDNAKNTLYLQMSSLRAEDTAVYYCTRESYSQGNYFDYWGQGTTVTVSS Sequence ID: 7

[0045] hu2F7 LCVR EIVMTQSPATLSLSPGERATLSCRASQSISDYLHWYQQKPGQSPRLLIKFASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQNGHSFSLTFGQGTKLEIK Sequence ID: 8

[0046] hu2F7 HC EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYYMSWVRQAPGKGLEWVAYVSSGGGSTYYSDSVKGRFTISRDNAKNTLYLQMSSLRAEDTAVYYCTRESYSQGNYFDYWGQ GTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence ID: 9

[0047] hu2F7 LC EIVMTQSPATLSLSPGERATLSCRASQSISDYLHWYQQKPGQSPRLLIKFASQSISGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQNGHSFSLTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID: 10

[0048] 2) Anti-B7-H4 antibody-drug conjugate (Compound 34, Shanghai Hansoh BioMedical Co., Ltd.) is disclosed in WO2020244657 and is manufactured according to the manufacturing method of Example 9 of WO2020244657.

[0049] Compound 34 has the following structure. [ka]

[0050] Antibody content C mab By measuring the absorbance at 280 nm, the low molecular weight content of C is determined. Drug All measurements were based on a standard calibration curve, using absorbance at 370 nm. The mean drug-antibody ratio y=C Drug / C mab Using the method described above, the average DAR of the exemplified product, hu2F7-exatecan (compound 34), was measured to be 6.1.

[0051] 2.3 Equipment [Table 3]

[0052] 2.4 Reagents [Table 4]

[0053] 3. Experimental Procedures and Data Processing 3.1 Animals We purchased NU / NU mice aged 42-62 days from Zhejiang Vitalriver Laboratory Animal Technology Co., Ltd.

[0054] 3.2 Animal Models Human cholangiocarcinoma tumor tissue LD1-0060-202214 was evenly cut into pieces approximately 3mm x 3mm x 3mm (20-30mg) in size and transplanted subcutaneously into the right flank of NU / NU mice.

[0055] 3.3 Tumor measurement and grouping of tumor-carrying mice Before starting treatment, the body weight of all animals was measured, and the tumor volume was measured using calipers as follows: Analytical value (mm²) 3) = length (mm) × width (mm) × width (mm) / 2. Mice with tumors were randomly grouped based on body weight and tumor volume.

[0056] 3.4 Treatment of tumor-carrying mice Each group was administered the test drug (method of administration: tail vein injection, dose: 10 mL / kg; frequency of administration: single dose; administration cycle: 21 days; solvent: PBS solution). After the start of test drug administration, tumor size and body weight were measured twice a week, and the animals were euthanized after the experiment was completed.

[0057] [Table 5]

[0058] 3.5 Data Processing The data were analyzed using GraphPad Prism and similar statistical software. The antitumor effect of the compounds was evaluated using TGI (%) and ΔT / ΔC (%). ΔT / ΔC (%) = (T-T0) / (C-C0) × 100, where T and C represent the tumor volume of the treatment and control groups at the end of the experiment, respectively, and T0 and C0 represent the tumor volume of the treatment and control groups at the start of the experiment, respectively. Tumor growth inhibition (TGI) (%) = 100 - ΔT / ΔC (%). Tumor regression was calculated as TGI (%) = 100 - (T-T0) / T0 × 100.

[0059] 4. Experimental Results Table 3 shows the tumor growth inhibitory effect of anti-B7-H4 antibody-drug conjugates on subcutaneous xenografts of human cholangiocarcinoma LD1-0060-202214. [Table 6]

[0060] All test drugs were administered intravenously as a single dose, and the animals were observed for three consecutive weeks. On the final day of the experiment (day 21), the mean tumor volume in tumor-carrying mice in the carrier control group, the hu2F7 unbound antibody 10 mg / kg administration group, and the compound 34 10 mg / kg administration group was 470.26 ± 73.86 mm², respectively. 3 525.97±71.92mm 3 and 216.02±38.40mm 3 Compared to the carrier control group, the hu2F7 unbound antibody 10 mg / kg administration group and the compound 34 10 mg / kg administration group showed tumor growth inhibition (TGI) (%) of -16.11% and 72.95%, respectively, and △T / △C (%) of 116.11% and 27.05%, respectively. Statistical data showed that the mean tumor volume was statistically significantly reduced in the compound 34 administration group compared to the carrier group (p<0.05).

[0061] 5. Experimental Conclusions In embodiments of the present invention, the anti-B7-H4 antibody-drug conjugate (i.e., compound 34) demonstrated a significant tumor-suppressing effect. After a single tail vein injection and 3 weeks of observation, compound 34 significantly inhibited tumor growth in a xenograft model of human cholangiocarcinoma LD1-0060-202214 in nude mice. No significant weight loss or other treatment-related adverse events were observed.

[0062] While the present invention has been described above with reference to various embodiments, it should be understood that the present invention is not limited to these embodiments. The present invention encompasses the general principles disclosed herein, and those skilled in the art can make changes or modifications without departing from the spirit and scope of the invention. Therefore, this specification is provided as an example and is not intended to be limiting.

Claims

1. The use of an anti-B7-H4 antibody-drug conjugate, or a pharmaceutically acceptable salt thereof, stereoisomer, metabolite, or solvent compound thereof, in the manufacture of a drug for the prevention and / or treatment of cancer, wherein the anti-B7-H4 antibody-drug conjugate is of formula (I): 【Chemistry 1】 [In the formula, y represents 1 to 20, preferably y represents 2 to 8, more preferably y represents 4 to 6 or 7 to 8; y is a decimal or an integer, even more preferably y represents 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3 or 6.4; Ab represents an anti-B7-H4 antibody or its antigen-binding fragment, which includes a variable region of the antibody heavy chain and a variable region of the antibody light chain. The variable region of the antibody heavy chain includes HCDR1, HCDR2, and HCDR3 as shown in SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively; and the variable region of the antibody light chain includes LCDR1, LCDR2, and LCDR3 as shown in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, respectively. Having a structure represented by; and The aforementioned use is for cancer in which the expression of B7-H4 is low.

2. The use according to claim 1, wherein the B7-H4 antibody or its antigen-binding fragment is a humanized antibody or a fragment thereof.

3. The use according to claim 2, wherein the humanized antibody comprises the heavy chain variable region shown in SEQ ID NO: 7 and the light chain variable region shown in SEQ ID NO: 8; preferably, the humanized antibody comprises the heavy chain shown in SEQ ID NO: 9 and the light chain shown in SEQ ID NO:

10.

4. The use according to any one of claims 1 to 3, wherein the cancer with low B7-H4 expression has a B7-H4 expression level of 1+ or 2+ by immunohistochemistry (IHC).

5. The use according to claim 4, wherein the cancer with low B7-H4 expression is cholangiocarcinoma with low B7-H4 expression, and preferably cholangiocarcinoma with a B7-H4 expression level of IHC 1+ or 2+.

6. The use according to any one of claims 1 to 5, wherein the prevention and / or treatment comprises administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, to a subject in need, at a dose of 0.5 to 15 mg / kg body weight, preferably 1 to 10 mg / kg body weight, and more preferably 3 to 10 mg / kg body weight.

7. The use according to any one of claims 1 to 5, wherein the prevention and / or treatment comprises administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, to a subject in need, via an oral, parenteral, or transdermal route, wherein the parenteral route is selected from intravenous, subcutaneous, or intramuscular injection.

8. The use according to any one of claims 1 to 5, wherein the prevention and / or treatment comprises administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, to a subject in need thereof, wherein the drug is in an injectable form, for example, a subcutaneous injection or an intravenous injection; the preferred injectable form is an injection or lyophilized powder for injection comprising the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, and a buffer, stabilizer, pH adjuster, and optionally present surfactant.

9. A method for preventing and / or treating cancer with low B7-H4 expression, The method comprises administering a therapeutically effective amount of the anti-B7-H4 antibody-drug conjugate of formula (I) described in any one of claims 1 to 3, or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, to a subject requiring it, preferably at a dose of 0.5 to 15 mg / kg body weight, more preferably 3 to 10 mg / kg body weight, or The procedure involves administering a therapeutically effective amount of a concomitant drug comprising the anti-B7-H4 antibody-drug conjugate of formula (I) as described in any one of claims 1 to 3, or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof. The aforementioned method.

10. A method for preventing and / or treating cancer with low B7-H4 expression according to claim 9, wherein the cancer with low B7-H4 expression is a cancer in which the B7-H4 expression level is IHC 1+ or 2+; preferably, the cancer with low B7-H4 expression is a cholangiocarcinoma with low B7-H4 expression; more preferably, the cancer with low B7-H4 expression is a cholangiocarcinoma in which the B7-H4 expression level is IHC 1+ or 2+.

11. A method for preventing and / or treating cancer with low B7-H4 expression according to claim 9, wherein an anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, is administered via an oral, parenteral, or transdermal route, wherein the parenteral route is selected from intravenous, subcutaneous, or intramuscular injection.

12. A method for preventing and / or treating cancer with low B7-H4 expression according to claim 9, wherein the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, is in an injectable form, for example, a subcutaneous injection or an intravenous injection; a more preferred injectable form is an injection or lyophilized powder for injection comprising the anti-B7-H4 antibody-drug conjugate of formula (I), or a pharmaceutically acceptable salt, stereoisomer, metabolite, or solvent compound thereof, and a buffer, stabilizer, pH adjuster, and optionally present surfactant.

13. A therapeutically effective amount of a drug formulation comprising an anti-B7-H4 antibody-drug conjugate of formula (I) according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, metabolite, or solvent compound thereof, further comprising a pharmaceutically acceptable carrier, wherein the anti-B7-H4 antibody-drug conjugate is administered at a dose of 0.5 to 15 mg / kg body weight, preferably 3 to 10 mg / kg body weight, and used for the treatment of cancer with low B7-H4 expression; preferably the cancer with low B7-H4 expression is cancer with a B7-H4 expression level of IHC 1+ or IHC 2+, more preferably the cancer with low B7-H4 expression is cholangiocarcinoma with low B7-H4 expression, and even more preferably the cancer with low B7-H4 expression is cholangiocarcinoma with a B7-H4 expression level of IHC 1+ or IHC 2+.

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