Method for treating gynecological malignant tumors by anti-HER2 antibody drug conjugate and application of anti-HER2 antibody drug conjugate

Abstract

The present disclosure relates to methods and uses of anti-HER2 antibody drug conjugates for treatment of gynecological malignant tumors. Specifically, the invention relates to application of the anti-HER2 antibody drug conjugate in preparation of drugs for treating gynecological malignant tumors expressed by HER2, and the structure of the antibody drug conjugate is shown as a formula (I). # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and relates to a method and use of an anti-HER2 antibody-drug conjugate (ADC) in treating gynecological malignancies. Background Art

[0002] HER2 is a member of the type I transmembrane tyrosine kinase receptor family. It is basically inactive in the monomeric state, but it can polymerize with the other three transmembrane tyrosine kinase members of the family, HER1, HER3, and HER4, leading to the phosphorylation of receptor tyrosine residues and the initiation of multiple signaling pathways (such as MAPK, PI3K / Akt, etc.), thereby promoting cell proliferation and tumor development. There are certain differences in HER2 expression or mutation in different tumor types. For example, breast cancer is highly dependent on HER2 signals, while gastric cancer is not as dependent on HER2 signals as breast cancer, and the intratumoral heterogeneity is high. Although both colorectal cancer and lung cancer have HER2 overexpression and mutation, the former has some overlap, while the latter basically does not overlap. Clinically, HER2 expression is closely related to patient prognosis. Breast cancer with high HER2 expression is highly invasive, has a short disease-free survival period, and has a poor prognosis. Due to the significant difference in the expression level of HER2 in normal cells and tumor cells, and its characteristics of having an extracellular domain, it has become an ideal target for tumor immunobiological therapy and is also one of the hot spots in the current research field of tumor treatment.

[0003] ADCs targeting HER2 work in the following ways: first, the antibody in the ADC specifically recognizes and binds to the HER2 receptor on the surface of the target cell, then enters the target cell through endocytosis, and releases the cytotoxic drug after decomposition inside the cell. Finally, the cytotoxic drug induces cell apoptosis by damaging DNA or acting on tubulin, thereby exerting an anti-tumor effect. If the cytotoxic drug has high membrane permeability, it may penetrate into the extracellular space after being released in the target cell, killing the surrounding HER2-negative cells. This effect is called the bypass killing effect. This effect may also occur if the cytotoxic drug is released before ADC endocytosis occurs. DS-8201a is a new generation of ADC drugs. In December 2019, DS-8201a was used to treat adult patients with unresectable or metastatic HER2-positive breast cancer who have previously received more than two HER2-targeted therapies. In May 2020, DS-8201a received breakthrough therapy designation from the FDA for the treatment of HER2-positive gastric cancer or gastroesophageal junction adenocarcinoma and HER2-mutated metastatic non-small cell lung cancer (NSCLC).

[0004] Although HER2 targeted drugs have made great progress in tumor treatment, they still face some challenges. In addition to breast cancer and gastric cancer, the HER2 overexpression rate of many other tumors is greater than 10%, such as gynecological malignancies including ovarian cancer, endometrial cancer, and cervical cancer, but there is a lack of standard treatment targeting HER2. Therefore, further research is needed to explore the efficacy of HER2 ADC drugs in gynecological malignancies with HER2 expression to meet clinical needs. Summary of the invention

[0005] The present disclosure provides uses and methods of an anti-HER2 antibody-drug conjugate in treating gynecological malignancies.

[0006] The present disclosure provides use of an anti-HER2 antibody-drug conjugate in preparing a drug for treating HER2-expressing gynecological malignancies.

[0007] The present disclosure provides anti-HER2 antibody drug conjugates for use in treating HER2-expressing gynecological malignancies.

[0008] The present disclosure provides a method for treating HER2-expressing gynecological malignancies, comprising administering a therapeutically effective amount of the anti-HER2 antibody drug conjugate to a subject.

[0009] In some embodiments, the HER2-expressing gynecological malignancy is selected from HER2-expressing advanced, recurrent and / or metastatic gynecological malignancies.

[0010] In some embodiments, the HER2-expressing gynecological malignancy is selected from any one of the following HER2-expressing tumors or any combination thereof: ovarian cancer, endometrial cancer, or cervical cancer.

[0011] In some embodiments, the HER2-expressing gynecological malignancy is selected from HER2-expressing advanced cervical cancer, recurrent ovarian cancer, or endometrial cancer.

[0012] In some embodiments, "recurrent ovarian cancer" refers to histopathologically confirmed recurrent epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer, which has been previously treated with a platinum-based regimen and has progressed or recurred < 6 months during or after the last platinum-containing treatment (completion of 4 or more courses of treatment); for platinum-resistant / platinum-refractory recurrent ovarian cancer, the patient has received no more than one line of systemic treatment.

[0013] In some embodiments, relapse or progression during the last platinum-containing treatment regimen or at the end of treatment must be clearly documented radiographic progression, and CA-125 progression can be used as a basis for determining the time of progression.

[0014] In some embodiments, "endometrial cancer" refers to patients with histopathologically confirmed endometrial cancer (excluding carcinosarcoma) who have experienced recurrence / metastasis after initial treatment; have received platinum-based systemic treatment (excluding radiosensitization chemotherapy) for the recurrent / metastatic stage, and have experienced disease recurrence or progression during treatment or after the end of treatment (receiving treatment for ≥4 cycles); and have had ≤2 lines of previous systemic treatment.

[0015] In some embodiments, "cervical cancer" refers to patients with histologically or cytologically confirmed cervical cancer who have recurred / metastasized after initial treatment; patients who have received platinum-based systemic treatment (excluding radiosensitization chemotherapy) for the recurrent / metastatic stage, and have experienced disease recurrence or progression during treatment or after the end of treatment (receiving treatment for ≥4 cycles); patients with ≤2 previous lines of systemic treatment; Note: Patients who received platinum-based systemic treatment (excluding radiosensitization chemotherapy) as neoadjuvant or adjuvant treatment, if the disease relapsed or progressed within 6 months during treatment or after the end of treatment (receiving treatment for ≥4 cycles), it was counted as first-line systemic treatment.

[0016] In some specific implementation schemes, patients who receive platinum-based systemic therapy (excluding radiosensitizing chemotherapy) as neoadjuvant or adjuvant therapy will be counted as first-line systemic therapy if the disease relapses or progresses during treatment or within 6 months after the end of treatment (≥4 cycles of treatment).

[0017] In some embodiments, the HER2-expressing gynecological malignancy is selected from any of the following HER2-expressing tumors: recurrent epithelial ovarian cancer, fallopian tube cancer or primary peritoneal cancer, recurrent or metastatic endometrial cancer, recurrent or metastatic cervical cancer.

[0018] In some embodiments, the HER2 detection standard refers to: ASCO / CAP gastric cancer, gastroesophageal junction adenocarcinoma HER2 interpretation standard 2016. In some embodiments, the HER2 expression is IHC 1+, 2+ or 3+. The central laboratory confirms HER2 expression: IHC 1+, 2+ or 3+; the subject can provide a specimen of the primary or metastatic tumor site for HER2 detection (paraffin block, paraffin embedded section or fresh tissue section); in some embodiments, subjects with IHC 1+ and IHC2+ need to test FISH.

[0019] In some embodiments, the anti-HER2 antibody drug conjugate is an anti-HER2 antibody drug conjugate selected from any one disclosed in WO2020063676 and WO2021190581, and the foregoing patents are incorporated into the present disclosure by reference.

[0020] In some embodiments, the anti-HER2 antibody drug conjugate is trastuzumab deruxtecan or trastuzumab emtansine

[0021] In some embodiments, the structure of the antibody drug conjugate is as shown in formula (I):

[0022]

[0023] in:

[0024] n is 3 to 8, and n is a decimal or an integer;

[0025] Pc is an anti-HER2 antibody or an antigen-binding fragment thereof.

[0026] In some embodiments, the anti-HER2 antibody drug conjugate has a structure as shown in the following formula:

[0027]

[0028] Wherein: n is 3 to 8, n is a decimal or an integer; Ab is an anti-HER2 antibody or an antigen-binding fragment thereof.

[0029] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof is selected from Trastuzumab, or an antigen-binding fragment thereof, Pertuzumab or an antigen-binding fragment thereof.

[0030] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof is Trastuzumab or an antigen-binding fragment thereof.

[0031] In some embodiments, the antibody drug conjugate has a structure as shown in the following formula:

[0032]

[0033] Here, n is 3 to 8 and is a decimal or an integer.

[0034] In an optional embodiment, n is 6 in the antibody-drug conjugate.

[0035] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively; and / or the light chain variable region comprises LCDR1 as shown in SEQ ID NO:4, LCDR2 sequence is SAS, and LCDR3 as shown in SEQ ID NO:5.

[0036] The aforementioned CDR sequences are shown in the following table:

[0037] Table 1. CDR sequences obtained by IMGT numbering rules

[0038] Antibody Anti-HER2 antibodies Serial Number Heavy chain CDR1 GFNIKDTY SEQ ID NO:1 Heavy chain CDR2 IYPTNGYT SEQ ID NO:2 Heavy chain CDR3 SRWGGDGFYAMDY SEQ ID NO:3 Light chain CDR1 QDV SEQ ID NO:4 Light chain CDR2 SAS Light chain CDR3 QQHYTTPPT SEQ ID NO:5

[0039] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises any one, any two, any three, any four, any five or six CDRs selected from HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 shown in SEQ ID NO: 1 to SEQ ID NO: 6.

[0040] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region comprises HCDR1, HCDR2, HCDR3 as shown in SEQ ID NO: 6, respectively, and the light chain variable region comprises LCDR1, LCDR2, LCDR3 as shown in SEQ ID NO: 7, respectively. The CDRs are defined according to the Kabat, IMGT, Chothia, AbM or Contact numbering systems. In some specific embodiments, the CDRs are defined according to the IMGT numbering system.

[0041] In some embodiments, the anti-HER2 antibody or its antigen-binding fragment is a murine antibody, a chimeric antibody, a humanized antibody, a human antibody, or an antigen-binding fragment of any of the foregoing. In some specific embodiments, or its antigen-binding fragment is selected from a humanized antibody or its antigen-binding fragment.

[0042] In some embodiments, the VH of the anti-HER2 antibody or antigen-binding fragment thereof comprises the amino acid sequence shown in SEQ ID NO: 6, or an amino acid sequence having at least 80%, 90% sequence identity thereto; and the VL comprises the amino acid sequence shown in SEQ ID NO: 7, or an amino acid sequence having at least 80%, 90% sequence identity thereto. Heavy chain variable region:

[0043] EVQLVESGGGLVQPGGSLRLSCAAS GFNIKDTY IHWVRQAPGKGLEWVAR IYPTNGYT

[0044] RYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYC SRWGGDGFYAMDY QUR

[0045] LVTVSS

[0046] SEQ ID NO:6 Light chain variable region:

[0047] DIQMTQSPSSLSASVGDRVTITCRAS QDV VAWYQQKPGKAPKLLIY SAS FLYSGVP

[0048] SRFSGSRSGTDFTLTISSLQPEDFATYYC QQHYTTPPT FQGKV

[0049] SEQ ID NO:7

[0050] In some embodiments, the anti-HER2 antibody or antigen-binding fragment thereof comprises any one or two of the above-mentioned VH and VL.

[0051] The above light chain variable region and light chain constant region sequences are combined to form a light chain sequence, and each heavy chain variable region and heavy chain constant region are combined to form a heavy chain sequence. An exemplary humanized antibody sequence is shown below:

[0052] Light chain:

[0053]

[0054] Heavy Chain:

[0055]

[0056] In some embodiments, the anti-HER2 antibody comprises a heavy chain and a light chain, wherein the heavy chain variable region comprises an amino acid sequence as shown in SEQ ID NO:9, or at least 80%, 90% identical thereto, and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO:10, or at least 80%, 90% identical thereto.

[0057] In some embodiments, the anti-HER2 antibody comprises any one or two of the aforementioned heavy chains and light chains.

[0058] In the context of the present disclosure, "at least 80%" and "at least 90%" encompass 80% and above, for example at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, and any numerical range therebetween.

[0059] In an optional embodiment, the single administration dose of the anti-HER2 antibody drug conjugate is about 1.0 mg / kg to about 10.0 mg / kg, for example, about 1.0 mg / kg to about 8.0 mg / kg, about 2.0 mg / kg to about 8.0 mg / kg, about 3.2 mg / kg to about 8.0 mg / kg, about 3.2 mg / kg to about 6.4 mg / kg, about 2.0 mg / kg to about 6.4 mg / kg.

[0060] In an optional embodiment, the anti-HER2 antibody drug conjugate is administered once a week, once every two weeks, once every three weeks, or once every four weeks.

[0061] In an optional embodiment, the single dose of the anti-HER2 antibody drug conjugate can be about 1.0 mg / kg, about 1.2 mg / kg, about 1.4 mg / kg, about 1.6 mg / kg, about 1.8 mg / kg, about 2.0 mg / kg, about 2.2 mg / kg, about 2.4 mg / kg, about 2.6 mg / kg, about 2.8 mg / kg, about 3.0 mg / kg, about 3.2 mg / kg, about 3.4 mg / kg, about 3.6 mg / kg, about 3.8 mg / kg, about 4.0 mg / kg, about 4.2 mg / kg, about 4.4 mg / kg, about 4.6 mg / kg, about 4.8 mg / kg, about 5.0 mg / kg, about 5.2 mg / kg , about 5.4 mg / kg, about 5.6 mg / kg, about 5.8 mg / kg, about 6.0 mg / kg, about 6.2 mg / kg, about 6.4 mg / kg, about 6.6 mg / kg, about 6.8 mg / kg, about 7.0 mg / kg, about 7.2 mg / kg, about 7.4 mg / kg, about 7.6 mg / kg, about 7.8 mg / kg, about 8.0 mg / kg, about 8.2 mg / kg, about 8.4 mg / kg, about 8.6 mg / kg, about 8.8 mg / kg, about 9.0 mg / kg, about 9.2 mg / kg, about 9.4 mg / kg, about 9.6 mg / kg, about 9.8 mg / kg or 10.0 mg / kg, and the dosage frequency is once every three weeks.

[0062] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 1.0 mg / kg, and the administration frequency is once every three weeks.

[0063] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 2.0 mg / kg, and the administration frequency is once every three weeks.

[0064] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 3.2 mg / kg, and the administration frequency is once every three weeks.

[0065] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 4.8 mg / kg, and the administration frequency is once every three weeks.

[0066] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 5.6 mg / kg, and the administration frequency is once every three weeks.

[0067] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 6.4 mg / kg, and the administration frequency is once every three weeks.

[0068] In an optional embodiment, the single dosage of the anti-HER2 antibody drug conjugate is about 8.0 mg / kg, and the administration frequency is once every three weeks.

[0069] In some embodiments, the administration route of the anti-HER2 antibody drug conjugate can be oral administration, parenteral administration, or transdermal administration, and the parenteral administration includes but is not limited to intravenous injection, subcutaneous injection, and intramuscular injection. In some specific embodiments, the administration route of the anti-HER2 antibody drug conjugate is intravenous injection.

[0070] In some embodiments, the anti-HER2 antibody drug conjugate is configured in an injectable form. Exemplarily, the injectable form of the antibody drug conjugate is an injection or a lyophilized powder injection, which comprises the antibody drug conjugate and one or more pharmaceutically acceptable excipients.

[0071] the term

[0072] In order to make the present disclosure more easily understood, certain technical and scientific terms are specifically defined below. Unless otherwise expressly defined herein, all other technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present disclosure belongs. Unless the context clearly requires otherwise, throughout the specification and claims, the words "comprising", "having", "including", etc. should be understood to have an inclusive meaning, rather than an exclusive or exhaustive meaning; that is, the meaning of "including but not limited to".

[0073] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and where it does not occur.

[0074] "About" and "approximately" refer to values ​​within an acceptable error range for a particular value as determined by a person of ordinary skill in the art, which value depends in part on how it is measured or determined (i.e., the limits of the measurement system). For example, "about" may mean within 1 or more than 1 standard deviation. Alternatively, "about" or "substantially including" may mean a range of up to 20%, such as between 1% and 15%, between 1% and 10%, between 1% and 5%, between 0.5% and 5%, between 0.5% and 1%, and in this disclosure, each case in which a number or numerical range is preceded by the term "about" also includes embodiments of the given number. Unless otherwise stated, when a specific value appears in the application and claims, the meaning of "about" or "substantially including" should be assumed to be within an acceptable error range for that specific value.

[0075] The term "and / or", such as "X and / or Y" should be understood to mean "X and Y" or "X or Y" and should be used to provide clear support for both meanings or either meaning.

[0076] "Antibody drug conjugate (ADC)" is to connect antibodies or antibody fragments with biologically active cytotoxins or small molecule drugs with cell-killing activity through stable chemical linker compounds, making full use of the antibody's specificity for tumor cell-specific or highly expressed antigens and the high efficiency of cytotoxins to avoid toxic side effects on normal cells. Compared with traditional chemotherapy drugs, antibody drug conjugates can accurately bind to tumor cells and reduce the impact on normal cells.

[0077] An antibody drug conjugate "retains its chemical stability" in a pharmaceutical formulation if the antibody drug conjugate does not show significant chemical changes. Chemical stability can be assessed by detecting and quantifying chemically altered forms of the protein. Degradation processes that often change the chemical structure of a protein include hydrolysis or truncation (assessed by methods such as size exclusion chromatography and CE-SDS), oxidation (assessed by methods such as peptide mapping in combination with mass spectrometry or MALDI / TOF / MS), deamidation (assessed by methods such as ion exchange chromatography, capillary isoelectric focusing, peptide mapping, isoaspartate measurement), and isomerization (assessed by measuring isoaspartate content, peptide mapping, etc.).

[0078] An antibody drug conjugate "retains its biological activity" in a pharmaceutical formulation if the biological activity of the antibody drug conjugate at a given time is within a predetermined range of the biological activity exhibited when the pharmaceutical formulation is prepared.

[0079] As used herein, the term "antibody" is used in the broadest sense, covering various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies; monospecific antibodies, multispecific antibodies (such as bispecific antibodies), full-length antibodies and antibody fragments (or antigen-binding fragments, or antigen-binding portions), as long as they exhibit the desired antigen-binding activity. Antibodies may refer to immunoglobulins, which are tetrapeptide chain structures formed by two identical heavy chains and two identical light chains connected by interchain disulfide bonds. The amino acid composition and arrangement order of the constant region of the heavy chain of immunoglobulins are different, so their antigenicity is also different. Based on this, immunoglobulins can be divided into five categories, or isotypes of immunoglobulins, namely IgM, IgD, IgG, IgA and IgE, and their corresponding heavy chains are μ chain, δ chain, γ chain, α chain and ε chain, respectively. The same class of Ig can be divided into different subclasses according to the difference in the amino acid composition of its hinge region and the number and position of the heavy chain disulfide bonds, such as IgG can be divided into IgG1, IgG2, IgG3, IgG4. Light chains are divided into κ chains or λ chains by different constant regions. Each of the five types of Ig can have a kappa chain or a lambda chain. The sequences of about 110 amino acids near the N-terminus of the antibody heavy chain and light chain vary greatly, which is the variable region (V region); the remaining amino acid sequences near the C-terminus are relatively stable, which is the constant region (C region). The variable region includes three hypervariable regions (CDRs) and four relatively conservative framework regions (FRs). The three hypervariable regions determine the specificity of the antibody, also known as the complementarity determining regions (CDRs). Each light chain variable region (VL) and heavy chain variable region (VH) consists of three CDR regions and four FR regions, arranged in the order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The three CDR regions of the light chain refer to LCDR1, LCDR2, and LCDR3; the three CDR regions of the heavy chain refer to HCDR1, HCDR2, and HCDR3.

[0080] For the determination or definition of CDRs, the deterministic depiction of CDRs and the identification of residues comprising the binding site of the antibody can be accomplished by resolving the structure of the antibody and / or resolving the structure of the antibody-ligand complex. This can be accomplished by any of the various techniques known to those skilled in the art, such as X-ray crystallography. A variety of analytical methods can be used to identify CDRs, including but not limited to the Kabat numbering system, the Chothia numbering system, the AbM numbering system, the IMGT numbering system, contact definitions, and conformational definitions.

[0081] The amino acid sequence boundaries of CDRs can be determined by various well-known schemes, for example: "Kabat" numbering convention (see Kabat et al. (1991), "Sequences of Proteins of Immunological Interest", 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD), "Chothia" numbering convention, "ABM" numbering convention, "contact" numbering convention (see Martin, ACR. Protein Sequence and Structure Analysis of Antibody Variable Domains [J]. 2001) and ImMunoGenTics (IMGT) numbering convention (Lefranc, MP et al., Dev. Comp. Immunol., 27, 55-77 (2003); Front Immunol. 2018 Oct 16; 9: 2278), etc.

[0082] The term "antigen-binding fragment" or "functional fragment" or "antigen-binding portion" refers to one or more fragments of an intact antibody that retains the ability to specifically bind to an antigen. It has been shown that fragments of full-length antibodies can be used to perform the antigen-binding function of an antibody. Exemplary examples of binding fragments encompassed by the term "antigen-binding fragment" include: (i) Fab fragment, a monovalent fragment consisting of VL, VH, CL and CH1 domains; (ii) F(ab')2 fragment, a bivalent fragment comprising two Fab fragments connected by a disulfide bridge on the hinge region, (iii) Fd fragment consisting of VH and CH1 domains; (iv) Fv fragment consisting of the VH and VL domains of a single arm of an antibody; (v) dsFv, a stable antigen-binding fragment formed by interchain disulfide bonds of VH and VL; (vi) scFv; (vii) diabodies comprising fragments such as scFv, dsFv, Fab, etc. "Homology" refers to the sequence similarity between two polynucleotide sequences or between two polypeptides. When the positions in the two compared sequences are occupied by the same base or amino acid monomer subunit, for example, if every position of the two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The percentage of homology between the two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared × 100. For example, when the sequences are optimally aligned, if 6 out of 10 positions in the two sequences are matched or homologous, then the two sequences are 60% homologous; if 95 out of 100 positions in the two sequences are matched or homologous, then the two sequences are 95% homologous. Typically, when aligning two sequences, the comparison is made to give the maximum percentage homology. For example, the comparison can be performed by the BLAST algorithm, in which the parameters of the algorithm are selected to give the maximum match between each sequence over the entire length of each reference sequence. The following references relate to the BLAST algorithm frequently used for sequence analysis: BLAST ALGORITHMS: Altschul, SF et al., (1990) J. Mol. Biol. 215: 403-410; Gish, W. et al., (1993) Nature Genet. 3: 266-272; Madden, TL et al., (1996) Meth. Enzymol. 266: 131-141; Altschul, SF et al., (1997) Nucleic Acids Res. 25: 3389-3402; Zhang, J. et al., (1997) Genome Res. 7: 649-656. Other conventional BLAST algorithms such as those provided by NCBI BLAST are also well known to those skilled in the art.

[0083] The term "linker unit" or "linking fragment" or "linking unit" refers to a chemical structure fragment or bond that is connected to an antibody or its antigen-binding fragment at one end and to a drug at the other end, and can also be connected to other linkers before being connected to the drug. The preferred embodiment of the present disclosure is represented by L and L 1 To L 4 , where L 1 The end is connected to the antibody, L 4 The end is connected to the structural unit Y and then connected to the compound or toxin.

[0084] The linker, including the extension, the spacer and the amino acid unit, can be synthesized by methods known in the art, such as those described in US2005-0238649A1. The linker can be a "cleavable linker" that facilitates the release of the drug in the cell. For example, an acid-labile linker (e.g., a hydrazone), a protease-sensitive (e.g., a peptidase-sensitive) linker, a photolabile linker, a dimethyl linker, or a disulfide-containing linker can be used (Chari et al., Cancer Research 52: 127-131 (1992); U.S. Patent No. 5,208,020).

[0085] The term "drug loading" refers to the average number of cytotoxic drugs loaded on each antibody or antigen-binding fragment thereof in the molecule of formula (I), and can also be expressed as the ratio of the amount of drug to the amount of antibody. The range of drug loading can be 0-12, preferably 1-10, more preferably 3-8, and most preferably 5.3-6.1 cytotoxic drugs (D) connected to each antibody or antigen-binding fragment thereof (Pc). In the embodiments of the present disclosure, the drug loading is expressed as n, which can be exemplified as the average of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. The average number of drugs per ADC molecule after the coupling reaction can be identified by conventional methods such as UV / visible light spectroscopy, mass spectrometry, ELISA experiments, and HPLC characteristics.

[0086] In one embodiment of the present disclosure, the cytotoxic drug is coupled to the N-terminal amino group, the ε-amino group of the lysine residue and / or the thiol group of the antibody or its antigen-binding fragment through a linker. Generally, the number of drug molecules that can be coupled to the antibody in the coupling reaction will be less than the theoretical maximum value.

[0087] The loading of cytotoxic drugs can be controlled by the following non-limiting methods, including:

[0088] (1) Control the molar ratio of the linking reagent and the monoclonal antibody,

[0089] (2) Control the reaction time and temperature,

[0090] (3) Select different reaction reagents.

[0091] The preparation of conventional pharmaceutical compositions is described in the Chinese Pharmacopoeia.

[0092] "Administering" and "treating" as applied to an animal, a human, a laboratory subject, a cell, a tissue, an organ, or a biological fluid, refers to the contact of an exogenous drug, therapeutic agent, diagnostic agent, or composition with an animal, a human, a subject, a cell, a tissue, an organ, or a biological fluid. "Administering" and "treating" can refer to, for example, treatment, pharmacokinetics, diagnosis, research, and experimental procedures. Treatment of cells includes contact of an agent with a cell, and contact of an agent with a fluid, wherein the fluid is in contact with the cell. "Administering" and "treating" also mean in vitro and ex vivo treatment of, for example, a cell by an agent, a diagnostic, a combination composition, or by another cell. "Treatment" as applied to humans, veterinary medicine, or research subjects refers to therapeutic treatment, prophylactic or preventative measures, research and diagnostic applications.

[0093] "Treatment" means administering an internal or external therapeutic agent, such as a composition comprising any of the binding compounds of the present disclosure, to a patient who has one or more symptoms of a disease for which the therapeutic agent is known to have a therapeutic effect. Typically, the therapeutic agent is administered in an amount effective to alleviate one or more symptoms of the disease in the treated patient or population to induce regression of such symptoms or inhibit the development of such symptoms to any clinically measurable degree. The amount of therapeutic agent effective to alleviate any specific disease symptom (also referred to as a "therapeutically effective amount") may vary according to a variety of factors, such as the patient's disease state, age, and weight, and the ability of the drug to produce the desired therapeutic effect in the patient. Whether the disease symptom has been alleviated can be evaluated by any clinical test method commonly used by physicians or other professional health care personnel to evaluate the severity or progression of the symptom. Although embodiments of the present disclosure (e.g., treatment methods or articles of manufacture) may not be effective in alleviating every symptom of the target disease, they should alleviate the target disease symptoms in a statistically significant number of patients as determined by any statistical test known in the art, such as Student's t-test, chi-square test, U test according to Mann and Whitney, Kruskal-Wallis test (H test), Jonckheere-Terpstra test, and Wilcoxon test.

[0094] An "effective amount" includes an amount sufficient to ameliorate or prevent a symptom or condition of a medical disease. An effective amount also means an amount sufficient to allow or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's general health, the method, route and dosage of administration, and the severity of side effects. An effective amount may be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

[0095] In the anti-HER2 antibody drug conjugate of the present disclosure, "n" refers to the average number of cytotoxic drugs loaded on each antibody or antigen-binding fragment thereof in the antibody drug conjugate molecule, and can also be expressed as the ratio of the amount of drug to the amount of antibody, which is the average number of drugs per ADC molecule after the coupling reaction is identified by hydrophobic chromatography (HIC) mass spectrometry. DETAILED DESCRIPTION

[0096] The present disclosure is further described below in conjunction with examples, but these examples are not intended to limit the scope of the present disclosure. Experimental methods in the examples of the present disclosure that do not specify specific conditions are usually carried out under conventional conditions, such as the Antibody Technology Laboratory Manual and the Molecular Cloning Manual published by the Cold Spring Harbor Laboratory; or under the conditions recommended by the raw material or product manufacturer. Reagents that do not specify specific sources are conventional reagents purchased from the market.

[0097] Example 1. Preparation of anti-HER2 antibody drug conjugates

[0098] According to the production method described in WO2021190581A, Trastuzumab (anti-HER2 antibody) and exotecan analogs were used to prepare the anti-HER2 antibody-drug conjugate shown in the following structure, and the average value was calculated by the HIC method: n=6.0, i.e., ADC-32.

[0099]

[0100] The following is the sequence of Trastuzumab:

[0101] Light chain:

[0102]

[0103] Heavy Chain:

[0104]

[0105] Note: The underlined part is the CDR region, and the italic part is the constant region.

[0106] Example 2. An open, international, multicenter phase I clinical study of the safety, tolerability, pharmacokinetics, and efficacy of an anti-HER2 antibody drug conjugate for injection in patients with advanced solid tumors expressing or mutated HER2

[0107] 1. Experimental drugs

[0108] The anti-HER2 antibody drug conjugate prepared in Example 1, lyophilized powder injection, specification: 100 mg / bottle.

[0109] 2. Research methods and results

[0110] The study enrolled 6 subjects with gynecological malignancies. The preliminary efficacy results are shown below, showing certain anti-tumor activity, which supports further exploration.

[0111]

[0112]

[0113] Example 3. Open, multicenter phase II clinical study of anti-HER2 antibody-drug conjugates for injection in the treatment of HER2-expressing gynecological malignancies

[0114] 1. Experimental drugs

[0115] The anti-HER2 antibody drug conjugate prepared in Example 1, lyophilized powder injection, specification: 100 mg / bottle.

[0116] 2. Enrolled subjects

[0117] 1). Patients voluntarily participated in this study and signed the informed consent form;

[0118] 2). 18 to 75 years old;

[0119] 3) The study enrolled patients with advanced cervical cancer, recurrent ovarian cancer and endometrial cancer:

[0120] Cohort 1 recurrent ovarian cancer: histopathologically confirmed recurrent epithelial ovarian cancer, fallopian tube cancer, or primary peritoneal cancer, previously treated with a platinum-based regimen, with progression or recurrence < 6 months during or after the last platinum-containing treatment (completion of 4 or more courses of treatment); for platinum-resistant / refractory recurrent ovarian cancer, received no more than first-line systemic treatment.

[0121] Note: Relapse or progression during the last platinum-containing treatment regimen or at the end of treatment must be clearly documented radiographic progression, and CA125 progression can be used as a basis for determining the time of progression.

[0122] Cohort 2 Endometrial cancer: patients with histopathologically confirmed endometrial cancer (excluding carcinosarcoma) who have recurred / metastasized after initial treatment; patients who have received platinum-based systemic treatment (excluding radiosensitization chemotherapy) for the recurrent / metastatic stage, and have experienced disease recurrence or progression during or after treatment (≥4 cycles); patients with ≤2 previous lines of systemic treatment;

[0123] Cohort 3 cervical cancer: patients with histologically or cytologically confirmed cervical cancer who have recurred or metastasized after initial treatment; patients who have received platinum-based systemic treatment (excluding radiosensitization chemotherapy) for the recurrent and metastatic stage, and have experienced disease recurrence or progression during or after treatment (≥4 cycles); patients with ≤2 previous lines of systemic treatment;

[0124] Note: Neoadjuvant or adjuvant therapy (excluding chemotherapy for radiosensitization) is counted as first-line systemic treatment if the disease recurs or progresses within 1 year after initial standard surgery or within 6 months after radiotherapy.

[0125] 4). The central laboratory confirms HER2 expression: IHC1+, 2+ or 3+; the subject can provide specimens of primary or metastatic tumors for HER2 testing (paraffin blocks, paraffin-embedded sections or fresh tissue sections are acceptable). Note: The scoring criteria for HER2 testing are determined by the central laboratory.

[0126] 5) At least one measurable lesion that meets the RECISTv1.1 standard (RECISTv1.1 requires that the long diameter of the measurable lesion on spiral CT scan be ≥10mm or the short diameter of the enlarged lymph node be ≥15mm);

[0127] 6).ECOG score: 0-1 points;

[0128] 7). Expected survival period ≥ 12 weeks;

[0129] 8) The functions of important organs meet the following requirements (no blood components or cell growth factor correction treatment was used within 14 days before the first medication):

[0130] Absolute neutrophil count ≥ 1.5 × 10 9 / L;

[0131] Platelets≥100×10 9 / L;

[0132] Hemoglobin ≥90 g / L;

[0133] Serum albumin ≥30 g / L;

[0134] Total bilirubin ≤1.5×ULN;

[0135] Prothrombin time and partial thromboplastin time ≤1.5×ULN;

[0136] ALT and / or AST ≤ 2.5 × ULN (for patients with liver metastasis, ALT and / or AST ≤ 5 × ULN);

[0137] Serum creatinine ≤1.5×ULN or creatinine clearance ≥50 mL / min

[0138] Female QTcF≤470msec;

[0139] The left ventricular ejection fraction (LVEF) of the heart should be ≥50%. The results of cardiac function tests should be completed within 28 days before the first use of the drug.

[0140] 3. Method of administration

[0141] Anti-HER2 antibody-drug conjugate: administered by intravenous drip, once every 3 weeks (Q3W), dosage (6.4 mg / kg and 4.8 mg / kg, the lowest allowed adjustment to 3.2 mg / kg).

[0142] 4. Result evaluation

[0143] Effectiveness Analysis:

[0144] The data are from the 101 study gynecological tumor cohort (cutoff date 2024 / 05 / 13) and the first phase of the 209 study (cutoff date 2024 / 05 / 15). A total of 99 subjects with advanced gynecological tumors were enrolled in the 4.8 and 6.4 mg / kg dose groups, of which 81 subjects with platinum-resistant recurrent ovarian cancer or endometrial cancer or cervical cancer that progressed after 1-2 lines of systemic treatment were enrolled in the target dose (4.8 mg / kg dose group); a total of 63 subjects completed at least one efficacy evaluation. Table 1 shows the preliminary efficacy results of the anti-HER2 antibody drug conjugate 4.8 mg / kg monotherapy for HER2-expressing gynecological malignancies.

[0145] Table 1

[0146]

[0147] Data cutoff date: 101 studies 2024 / 05 / 13, 209 studies 2024 / 05 / 15. Percentages are calculated with N as the denominator and only when the numerator is not zero.

[0148] *Only subjects who used the trial drug at least once and had a baseline tumor assessment and at least one efficacy assessment after baseline were counted.

[0149] #The response rate includes subjects with CR, confirmed PR and PR to be confirmed.

[0150] At the target dose of 4.8 mg / kg of anti-HER2 antibody-drug conjugate, the response rates (including responses to be confirmed) of monotherapy for platinum-resistant ovarian cancer, endometrial cancer and cervical cancer that progressed after 1-2 lines of treatment were 50.0%, 50.0% and 47.1%, respectively, and the disease control rates (DCR) were 88.2%, 83.3% and 94.1%, respectively; in the HER2 IHC 2+ / 3+ subgroup, the response rates (including responses to be confirmed) of platinum-resistant ovarian cancer, endometrial cancer and cervical cancer that progressed after 1-2 lines of treatment were 53.6%, 50.0% and 45.5%, respectively.

[0151] Security Analysis:

[0152] As of May 15, 2024, the common treatment-related adverse events (TRAEs) of 71 subjects at the RP2D 4.8mg / kg dose level of anti-HER2 antibody drug conjugates were (incidence ≥ 20%): decreased neutrophil count, anemia, decreased white blood cell count, decreased platelet count, decreased lymphocyte count, nausea, vomiting, decreased appetite, and alopecia. The incidence of TEAEs of CTCAE>=Grade 3 was 64.8%.

Claims

1. Use of an anti-HER2 antibody-drug conjugate in the preparation of a drug for treating HER2-expressing gynecological malignancies, wherein the structure of the antibody-drug conjugate is shown in formula (I): in: n is 3 to 8, and n is a decimal or an integer; Pc is an anti-HER2 antibody or an antigen-binding fragment thereof.

2. The use according to claim 1, wherein the anti-HER2 antibody or its antigen-binding fragment is selected from Trastuzumab, Pertuzumab, or an antigen-binding fragment thereof; preferably Trastuzumab or an antigen-binding fragment thereof.

3. The use according to claim 1, wherein The anti-HER2 antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively, and the VL comprises LCDR1 as shown in SEQ ID NO:4, LCDR3 as shown in SEQ ID NO:5, and the LCDR2 sequence is SAS; Preferably, the VH comprises an amino acid sequence as shown in SEQ ID NO:6, or having at least 90% identity thereto, and the VL comprises an amino acid sequence as shown in SEQ ID NO:7, or having at least 90% identity thereto; Preferably, the anti-HER2 antibody comprises a heavy chain comprising an amino acid sequence as shown in SEQ ID NO:8 or having at least 90% identity thereto, and a light chain comprising an amino acid sequence as shown in SEQ ID NO:9 or having at least 90% identity thereto.

4. The use according to any one of claims 1 to 3, wherein the anti-HER2 antibody drug conjugate has a structure as shown in the following formula: in, n is 3 to 8, and n is a decimal or an integer.

5. The use according to any one of claims 1 to 4, wherein the single dose of the anti-HER2 antibody drug conjugate is about 1.0 mg / kg to about 10.0 mg / kg, preferably about 1.0 mg / kg, about 2.0 mg / kg, about 3.2 mg / kg, about 4.8 mg / kg, about 5.6 mg / kg, about 6.4 mg / kg or about 8.0 mg / kg, and the frequency of administration is once a week, once every two weeks, once every three weeks or once every four weeks.

6. The use according to any one of claims 1 to 5, wherein The HER2-expressing gynecological malignancies are selected from HER2-expressing advanced, recurrent and / or metastatic gynecological malignancies.

7. The use according to any one of claims 1 to 6, wherein The HER2-expressing gynecological malignant tumor is selected from any one or any combination of the following tumors expressing HER2: ovarian cancer, endometrial cancer, and cervical cancer.

8. The use according to any one of claims 1 to 7, wherein The HER2-expressing gynecological malignancy is selected from HER2-expressing advanced cervical cancer, recurrent ovarian cancer or endometrial cancer.

9. The use according to any one of claims 1 to 8, wherein The HER2 expression is IHC 1+, 2+ or 3+.

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