Combination therapy including anti-HER2 bispecific antibodies and its use

Abstract

This disclosure relates to the biopharmaceutical field, and more specifically to combination therapeutic agents containing anti-HER2 bispecific antibodies and their use, particularly to the use of anti-HER2 bispecific antibodies in combination with docetaxel albumin compositions in the preparation of drugs for the treatment of advanced malignant solid tumors.

Description

[Technical Field]

[0001] The present invention relates to the biopharmaceutical field, and more specifically to combination therapeutic agents containing an anti-HER2 bispecific antibody and their use, particularly to the use of an anti-HER2 bispecific antibody in combination with a docetaxel albumin composition in the preparation of drugs for treating advanced malignant solid tumors. [Background technology]

[0002] The latest data released by the International Agency for Research on Cancer shows that in 2020, there were 2.26 million new cases of breast cancer worldwide, surpassing lung cancer to become the leading cause of new cancer cases globally, with 684,000 deaths, making it the leading cause of cancer death in women. In particular, China saw 416,000 new breast cancer cases, accounting for 18.4% of the world's new cases, and 117,000 deaths, accounting for 17.1% of the world's breast cancer deaths. These cases are on an upward trend year by year, posing a serious threat to the lives and health of women in China, and making it one of the key priorities in tumor prevention and treatment in China.

[0003] Based on differences in estrogen receptor / progesterone receptor and human epidermal growth factor receptor 2 (HER2) expression, breast cancer is classified into different histopathological subtypes. Among these, HER2-positive breast cancer accounts for approximately 15-20% of all breast cancers, and the proportion of HER2-positive breast cancer is higher in China (20-30%) compared to Western countries. HER2-positive breast cancer is a more malignant subtype of breast cancer with a younger population. Compared to hormone receptor-positive / HER2-negative and triple-negative breast cancers, HER2-positive breast cancer has a higher proportion of patients at stage III-IV at initial diagnosis, a higher histological grade, and the majority of patients are under 65 years of age at initial diagnosis. Currently in China, trastuzumab, pertuzumab, lapatinib, neratinib, pirotinib, and trastuzumab emtansine (T-DM1) are approved for the treatment of HER2-positive breast cancer. These drugs cover the entire treatment process for HER2-positive breast cancer, including neoadjuvant therapy, adjuvant therapy after surgery, and systemic treatment for recurrent and metastatic cancer. Nevertheless, palliative salvage therapy remains the primary goal of treatment for recurrent and metastatic breast cancer, and further improvements are expected.

[0004] In China, approximately 20% of patients have advanced breast cancer (stage III / IV) at their initial diagnosis. Additionally, about 10% of HER2-positive early-stage patients experience disease recurrence three years after curative surgery. First-line treatment with trastuzumab, pertuzumab, and docetaxel significantly improves PFS and OS in patients with HER2-positive metastatic breast cancer. However, the majority of patients still experience disease progression after treatment, indicating a continued need for more effective new treatment options. Furthermore, the combination therapy of trastuzumab with pertuzumab and docetaxel requires periodic administration until disease progression or intolerance occurs. However, each cycle (3 weeks) requires pertuzumab to be infused intravenously for 30-60 minutes (the loading dose requires infusion for 60 minutes), and trastuzumab to be infused intravenously for 30-90 minutes (the loading dose requires infusion for 90 minutes). Additionally, a 30-60 minute observation period is recommended between the administrations of both drugs. This long-term, repeated intravenous infusion treatment is inconvenient, increases the length of hospital stays for patients, increases the risk of hospital-acquired infections, and occupies more medical resources, so further solutions are needed. In summary, HER2-positive breast cancer urgently needs a more effective, safe, and convenient first-line treatment option.

[0005] Docetaxel is obtained by the semi-synthesis of a non-cytotoxic precursor (10-deacetylbaccatin III) extracted from yew needles and is a paclitaxel analog with stronger antitumor activity compared to paclitaxel. Docetaxel has relatively low water solubility, and the currently available commercially available formulation is a standard injectable solution, which was first developed by Sanofi under the trade name TAXOTERE (registered trademark). The usual dosage is administered once every three weeks by intravenous injection at a dose of 75 mg / m² over 1 hour. 2However, docetaxel injections, such as Taxotere®, contain ethanol and Tween80, which can cause severe allergic reactions, require dexamethasone pretreatment, result in poor patient compliance, and ethanol affects the central nervous system, necessitating a reduction in the infusion rate to alleviate symptoms of toxicity. [Overview of the Initiative]

[0006] In response to the shortcomings of the prior art, the present invention provides a combination drug for treating cancer, comprising an anti-HER2 bispecific antibody and a docetaxel albumin composition for the treatment of HER2-positive recurrent metastatic breast cancer. This combination drug has superior antitumor therapeutic efficacy compared to conventional standard treatment plans, offers good overall safety, and is more convenient to administer, providing such patients with a first-line treatment option that is more effective, safer, and more convenient to administer.

[0007] In a first embodiment, the present invention provides the use of an anti-HER2 antibody and a second therapeutic agent in the preparation of a drug for treating cancer, wherein the second therapeutic agent is a docetaxel albumin composition.

[0008] In a second embodiment, the present invention provides the use of an anti-HER2 antibody in the preparation of a drug for enhancing the therapeutic effect of a second therapeutic agent for treating cancer, wherein the second therapeutic agent is a docetaxel albumin composition.

[0009] In a third embodiment, the present invention provides a method for treating cancer with an anti-HER2 antibody and a second therapeutic agent, wherein the second therapeutic agent is a docetaxel albumin composition.

[0010] In a fourth aspect, the present invention provides a pharmaceutical composition, reagent kit, or drug kit for treating cancer comprising an anti-HER2 antibody and a second therapeutic agent, wherein the second therapeutic agent is a docetaxel albumin composition.

[0011] Each of the above aspects: In some embodiments, the combination of the anti-HER2 antibody and the second therapeutic agent is synergistic in the treatment of cancer.

[0012] In some embodiments, the anti-HER2 antibody is an anti-HER2 bispecific antibody.

[0013] In some embodiments, the anti-HER2 antibody is a bispecific antibody that targets different epitopes of the human HER2 protein.

[0014] In some embodiments, the anti-HER2 antibody is a bispecific antibody that targets different epitopes of the human HER2 protein, namely domain II and domain IV.

[0015] In some embodiments, the anti-HER2 antibody is a recombinant humanized antibody. In some embodiments, the anti-HER2 antibody contains two common light chains having the same sequence. In some embodiments, the common light chain includes CDR-L1 shown in SEQ ID NO: 1, CDR-L2 shown in SEQ ID NO: 2, and CDR-L3 shown in SEQ ID NO: 3. In some embodiments, the common light chain includes a light chain variable region shown in SEQ ID NO: 4, or the light chain variable region consists of the amino acid sequence shown in SEQ ID NO: 4. In some embodiments, the common light chain includes or consists of the amino acid sequence shown in SEQ ID NO: 5.

[0016] In some embodiments, the common light chain of the anti-HER2 antibody is a Kappa subtype.

[0017] In some embodiments, the anti-HER2 antibody comprises two heavy chains with different sequences. In some embodiments, the heavy chain comprises a heavy chain I that targets the HER2 protein domain II. In some embodiments, the heavy chain I comprises CDR-H1-1 shown in SEQ ID NO: 6, CDR-H1-2 shown in SEQ ID NO: 7, and CDR-H1-3 shown in SEQ ID NO: 8. In some embodiments, the heavy chain I comprises the heavy chain I variable region shown in SEQ ID NO: 9, or the heavy chain I variable region consists of the amino acid sequence shown in SEQ ID NO: 9. In some embodiments, the heavy chain I comprises or consists of the amino acid sequence shown in SEQ ID NO: 10 or SEQ ID NO: 16.

[0018] In some embodiments, the anti-HER2 antibody comprises two heavy chains with different sequences. In some embodiments, the heavy chain comprises a heavy chain II that targets the HER2 protein domain IV. In some embodiments, the heavy chain II comprises CDR-H2-1 shown in SEQ ID NO: 11, CDR-H2-2 shown in SEQ ID NO: 12, and CDR-H2-3 shown in SEQ ID NO: 13. In some embodiments, the heavy chain II comprises the heavy chain II variable region shown in SEQ ID NO: 14, or the heavy chain II variable region consists of the amino acid sequence shown in SEQ ID NO: 14. In some embodiments, the heavy chain II comprises or consists of the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 17.

[0019] In some embodiments, the heavy chain comprises a heavy chain I that targets the HER2 protein domain II and a heavy chain II that targets the HER2 protein domain IV, where the heavy chain I and the heavy chain II are as described above.

[0020] In some embodiments, the anti-HER2 antibody is KN026 (hereinafter abbreviated as KN026) of Jiangsu Kangning Jerry Biopharmaceutical Co., Ltd.

[0021] In some embodiments, the cancer includes, but is not limited to, hematological tumors or solid tumors.

[0022] In some embodiments, the cancer is a hematological malignancy. In some embodiments, the cancer is lymphoma or leukemia. In some embodiments, the lymphoma or leukemia includes, but is not limited to, myeloma, B-cell lymphoma, mantle cell lymphoma, non-Hodgkin B-cell lymphoma, non-Hodgkin T-cell lymphoma, cutaneous lymphoma, anaplastic large cell lymphoma, multiple myeloma, indolent non-Hodgkin lymphoma, plasmacytoma, chronic lymphocytic leukemia, small lymphocytic lymphoma, follicular lymphoma. In some embodiments, the cancer is recurrent or refractory.

[0023] In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor includes, but is not limited to, bladder cancer, brain cancer, breast cancer, cervical cancer, thoracic tumor, endometrial cancer, esophageal squamous cell carcinoma, gastric cancer, head tumor, pancreatic cancer, cholangiocarcinoma, colorectal cancer, eye cancer, head and neck squamous cell carcinoma, urothelial cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, neck tumor, ovarian cancer, prostate cancer, testicular cancer, laryngeal cancer and uterine cancer, melanoma, skin cancer, salivary gland cancer, soft tissue sarcoma and osteosarcoma. In some embodiments, the cancer is recurrent or refractory.

[0024] In some embodiments, the cancer is selected from breast cancer.

[0025] In some embodiments, the breast cancer is HER2-positive breast cancer, preferably IHC3+ HER2-positive breast cancer or IHC2+ and ISH-positive HER2-positive breast cancer. In some embodiments, the breast cancer is selected from estrogen receptor-positive or -negative HER2-positive breast cancer, preferably HER2-positive breast cancer with both estrogen receptor and progesterone receptor negative, HER2-positive breast cancer with estrogen receptor and / or progesterone receptor positive.

[0026] In some embodiments, the breast cancer includes ductal carcinoma, lobular carcinoma, medullary carcinoma, colloid carcinoma, tubular carcinoma, inflammatory breast cancer, triple-negative breast cancer (TNBC).

[0027] In some embodiments, the breast cancer is recurrent or metastatic breast cancer. In some embodiments, the breast cancer is selected from breast cancer with organ metastasis or breast cancer without organ metastasis.

[0028] In some embodiments, the breast cancer is a recurrent and metastatic breast cancer.

[0029] In some embodiments, the breast cancer is recurrent or metastatic breast cancer that has not previously received systemic chemotherapy and / or HER2-targeted therapy for recurrent or metastatic breast cancer, or recurrent breast cancer that has recurred for 12 months or more after completion of neoadjuvant therapy, adjuvant therapy, or HER2-targeted therapy. The systemic chemotherapy is selected from taxane-based chemotherapy drugs such as docetaxel, albumin-paclitaxel, capecitabine, and vinorelbine, and the HER2-targeted therapy is trastuzumab, pertuzumab, pirotinib, or trastuzumab emtansine. The neoadjuvant therapy plan is selected from the conjugate (TDM-1), and the above-mentioned neoadjuvant therapy plan is selected from the combination or non-combination of trastuzumab, taxanes and pertuzumab with carboplatin, anthracyclines and cyclophosphamide and taxanes, trastuzumab, and pertuzumab. The above-mentioned adjuvant therapy plan is selected from the combination of anthracyclines and cyclophosphamide followed by a combination of taxanes and trastuzumab and pertuzumab, docetaxel and carboplatin and trastuzumab and pertuzumab, neratinib followed or not followed by trastuzumab / pertuzumab, and TDM-1.

[0030] In some embodiments, only the anti-HER2 antibody and the second therapeutic agent are considered therapeutic active ingredients.

[0031] In some embodiments, the uses, methods, pharmaceutical compositions, reagent kits, or drug kits described herein further comprise a third therapeutic agent, which is a chemotherapeutic agent. The chemotherapeutic agent is, for example, bendamustine, chlorambucil, cyclophosphamide, doxorubicin, vincristine, fludarabine, carboplatin, cisplatin, or any combination thereof.

[0032] In some embodiments, the uses, methods, pharmaceutical compositions, reagent kits, or drug kits described herein further comprise a therapeutic agent, such as granulocyte colony-stimulating factor (G-CSF).

[0033] In some embodiments, the anti-HER2 antibody and the second therapeutic agent are contained in the same formulation unit.

[0034] In some embodiments, the anti-HER2 antibody and the second therapeutic agent are each contained in separate formulation units.

[0035] In some embodiments, the drugs, pharmaceutical compositions, reagent kits, or drug kits described herein are administered to patients or subjects in need by a clinically acceptable method, preferably by intravenous infusion. Correspondingly, the anti-HER2 antibody and the second therapeutic agent are clinically acceptable formulations, preferably injectable dosage forms including liquid injections, injectable powders, and injectable tablets.

[0036] In several embodiments, the anti-HER2 antibody is prepared into a formulation, and in each formulation unit, the content of the anti-HER2 antibody or its pharmaceutically acceptable form (calculated by the amount of anhydrous anti-HER2 antibody) is in the range of approximately 0.01 mg to approximately 400 mg, preferably approximately 300 to 400 mg, more preferably approximately 300 mg, approximately 325 mg, approximately 350 mg, approximately 375 mg, and approximately 400 mg.

[0037] In some embodiments, the anti-HER2 antibody is administered in doses of approximately 0.01 mg / kg to 100 mg / kg, preferably approximately 20 to 40 mg / kg, approximately 20 to 30 mg / kg, approximately 25 to 35 mg / kg, and more preferably approximately 30 mg / kg.

[0038] In some embodiments, the anti-HER2 antibody is administered in one or more administration cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 or more administration cycles). Preferably, it is administered in three or more administration cycles. In some embodiments of the present invention, the administration cycles of the anti-HER2 antibody continue until the clinical benefit is lost (e.g., disease progression, resistance, death, or unacceptable toxicity is observed).

[0039] In some embodiments, the docetaxel albumin composition is prepared into a formulation in which the docetaxel content (calculated using anhydrous docetaxel) in each formulation unit is 50 to 100 mg, preferably 70 to 90 mg, and more preferably about 80 mg.

[0040] In some embodiments, the docetaxel albumin composition is preferably an injectable powder, and when calculated using anhydrous docetaxel, the active ingredient content is about 50 to 100 mg / injection, preferably about 70 to 90 mg / injection, and more preferably about 80 mg / injection.

[0041] In some embodiments, the docetaxel albumin composition is approximately 60-125 mg / m². 2 The concentration is preferably about 60-100 mg / m². 2 Comfortably approximately 60 mg / m² 2 , about 75 mg / m 2 Or approximately 100 mg / m² 2 It is administered in the following dosage.

[0042] In some embodiments, the docetaxel albumin composition is administered in one or more administration cycles (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 or more administration cycles). Preferably, it is administered in three or more administration cycles. In some embodiments of the present invention, the administration cycle of the docetaxel albumin composition continues until the clinical benefit is lost (e.g., disease progression, resistance, death, or unacceptable toxicity is observed).

[0043] In some embodiments of the present invention, the length of each cycle in one or more administration cycles is 14 days. In some embodiments of the present invention, the length of each cycle in one or more administration cycles is 21 days. In some embodiments of the present invention, the length of each cycle in one or more administration cycles is 28 days. Preferably, the length of each cycle in one or more administration cycles is 21 days.

[0044] In some embodiments of the present invention, the combination of the anti-HER2 antibody and / or docetaxel albumin is administered at a frequency of QW (once a week), Q2W (once every two weeks), Q3W (once every three weeks), or Q4W (once every four weeks).

[0045] In some embodiments of the present invention, the administration schedule for the anti-HER2 antibody and / or docetaxel albumin composition is once every three weeks (Q3W), with a treatment cycle of 21 days.

[0046] In some embodiments of the present invention, the administration schedule for the anti-HER2 antibody and / or docetaxel albumin composition is once every four weeks (Q4W), with a treatment cycle of 28 days.

[0047] In some embodiments of the present invention, the administration schedule for the anti-HER2 antibody and / or docetaxel albumin composition is once every two weeks (Q2W), with a treatment cycle of 14 or 28 days.

[0048] In some embodiments of the present invention, the administration schedule for the anti-HER2 antibody and / or docetaxel albumin composition is once a week (QW), administered for three consecutive weeks, followed by a one-week rest period, with a 28-day treatment cycle.

[0049] In some embodiments of the present invention, the administration schedule for the anti-HER2 antibody and / or docetaxel albumin composition is once a week (QW), administered for two consecutive weeks, followed by a one-week rest period, with a 21-day treatment cycle.

[0050] In some embodiments of the present invention, the anti-HER2 antibody and / or docetaxel albumin composition is administered on approximately day 1 of each administration cycle (for example, day 1, 2, 3, or 4).

[0051] In some embodiments of the present invention, the anti-HER2 antibody is administered before or after the docetaxel albumin composition.

[0052] The drugs, pharmaceutical compositions, reagent kits, or drug kits described herein are administered to patients or subjects in a therapeutically effective dose for the treatment of breast cancer. The breast cancer described above is as defined above.

[0053] In some embodiments, the drug, pharmaceutical composition, reagent kit, or drug kit of the present application comprises an anti-HER2 antibody, a docetaxel albumin composition, and is administered to a patient or subject in a therapeutically effective dose for the treatment of breast cancer, which is as described above.

[0054] In some embodiments, the anti-HER2 antibody is administered once every 2 to 4 weeks, preferably once every 3 weeks. On the first day of each cycle, it is administered at a dose of about 20 to 40 mg / kg, preferably about 25 to 35 mg / kg, more preferably about 30 mg / kg. For the first time, it is intravenously infused over 90 min ± 15 min, and for subsequent cycles, it is intravenously infused over 60 min ± 15 min. In some embodiments, the docetaxel albumin composition is administered once every 2 to 4 weeks, preferably once every 3 weeks. On the first day of each cycle, it is administered at a dose of about 60 to 125 mg / m 2 ², preferably about 60 to 100 mg / m 2 ², for example, about 60 mg / m 2 ², about 75 mg / m 2 ² or about 100 mg / m 2 ², and is intravenously infused over 60 min ± 10 min.

[0055] In some embodiments, the anti-HER2 antibody is administered once every 3 weeks. On the first day of each cycle, it is administered at a dose of about 30 mg / kg, and for the first time, it is intravenously infused over 90 min ± 15 min, and for subsequent cycles, it is intravenously infused over 60 min ± 15 min. In some embodiments, the docetaxel albumin composition is administered once every 3 weeks. On the first day of each cycle, it is administered at a dose of about 60 mg / m 2 ², about 75 mg / m 2 ² or about 100 mg / m 2 ², and is intravenously infused over 60 min ± 10 min.

[0056] In some embodiments, the docetaxel albumin composition described in the present application is prepared by referring to the method disclosed in WO2022184164A1. According to research, whether it is the suspension before lyophilization or the reconstituted suspension after lyophilization, it can be stable for at least 24 hours at room temperature conditions and at least 10 days under refrigerated conditions, indicating that turbidity of the suspension and sedimentation of nanoparticles do not occur. Compared with the case where a commercially available albumin-bound drug reconstituted suspension is stable for only 8 h, this product greatly reduces the clinical use limitations.

[0057] The docetaxel albumin composition described herein is a docetaxel albumin nanoparticle composition comprising docetaxel and acid-denatured albumin, and optionally comprising an osmotic pressure regulator and / or a pH regulator. The acid-denatured albumin is obtained by denaturing human serum albumin after adjusting the pH to an appropriate value by adding acid. Preferably, (1) The above acid is selected from acidic amino acids or acidic polypeptides, organic acids, and inorganic acids. The above acidic amino acids or acidic polypeptides include, but are not limited to, cysteine ​​hydrochloride and glutathione; the above organic acids include, but are not limited to, citric acid and tartaric acid; and the above inorganic acids include, but are not limited to, hydrochloric acid and sulfuric acid. The above acid is preferably cysteine ​​hydrochloride, glutathione, hydrochloric acid, more preferably cysteine ​​hydrochloride, and / or (2) The appropriate pH values ​​are preferably about 3.5 to 5.5, more preferably about 3.5 to 5.0, even more preferably about 3.8 to 4.7, and also preferably about 4.0 to 4.5, and / or (3) When calculated using anhydrous docetaxel, the mass ratio of docetaxel to human serum albumin is 1:(2.0~10.0), preferably 1:(3.0~7.0), more preferably 1:(4.0~6.0), most preferably about 1:4.5, and / or (4) The sodium octanoate content in the human serum albumin described above is 0.08 mmol / g protein or less, preferably 0.03 to 0.08 mmol / g protein, more preferably 0.04 to 0.08 mmol / g protein, and even more preferably 0.04 to 0.07 mmol / g protein.

[0058] In some embodiments, the particle size of the docetaxel albumin nanoparticles is about 60 to 200 nm, preferably about 90 to 150 nm, and more preferably about 90 to 135 nm.

[0059] The type of osmotic regulator is not particularly limited and may be selected from, for example, sodium chloride, glucose, phosphate, or citrate, with sodium chloride being preferred. In some embodiments, the osmotic regulator is sodium chloride, and the weight ratio of sodium chloride to docetaxel is (0.75-9):1, preferably (1-7):1, more preferably (1.5-4.5):1, and most preferably about 2.25:1.

[0060] The type of pH adjusting agent is not particularly limited and is used to adjust the pH value to an appropriate range. Preferably, the pH value range is 3.4 to 5.8, more preferably 3.6 to 5.6 or 3.8 to 5.0, more preferably 3.9 to 4.8, and most preferably about 4.1.

[0061] The docetaxel albumin composition described in this application does not require that the docetaxel be in hydrate form or not, and may be any one selected from anhydrous docetaxel, docetaxel hemihydrate, or docetaxel trihydrate, preferably anhydrous docetaxel. The docetaxel content described in this application is calculated using anhydrous docetaxel.

[0062] The drug described herein comprises a docetaxel albumin composition. The drug is in a clinically acceptable dosage form, preferably an injectable, more preferably a liquid injectable, or lyophilized powder injectable. For example, the docetaxel (albumin-bound) for injection used in a specific embodiment is also called DTX-HSA.

[0063] In some embodiments, the drug is a liquid injection, the pH of which is 3.4 to 5.8, preferably 3.6 to 5.6 or 3.8 to 5.0, more preferably 3.9 to 4.8. The particle size of the docetaxel albumin nanoparticles in the liquid injection is about 60 to 200 nm, preferably about 90 to 150 nm, more preferably about 90 to 135 nm, and the liquid injection contains 0 to 1.8% (w / v), preferably 0.45% to 1.8% (w / v), more preferably 0.9% to 1.8% (w / v), and even more preferably 0.9% (w / v) of sodium chloride. In some embodiments, the liquid injection contains 2 to 10 mg / mL, preferably 2 to 8 mg / mL of docetaxel.

[0064] In some embodiments, the drug is a lyophilized powder injection containing sodium chloride, where the weight ratio of sodium chloride to docetaxel is (0.75-9):1, preferably (1-7):1, more preferably (1.5-4.5):1, and most preferably about 2.25:1. The particle size of the docetaxel albumin nanoparticles is about 60-200 nm, preferably about 90-150 nm, and more preferably about 90-135 nm. The pH of the suspension used to prepare the lyophilized powder injection before lyophilization is 3.4-5.8, preferably 3.6-5.6 or 3.8-5.0, more preferably 3.9-4.8, and most preferably about 4.1.

[0065] The above lyophilized powder injection is reconstituted into a suspension using a redissolution medium before use. The redissolution medium is selected from water for injection, sodium chloride solution, or glucose solution, and is preferably water for injection. The pH of the resulting reconstituted suspension is 3.4 to 5.8, preferably 3.6 to 5.6 or 3.8 to 5.0, and more preferably 3.9 to 4.8. The particle size of the docetaxel albumin nanoparticles in the reconstituted suspension is about 60 to 200 nm, preferably about 90 to 150 nm, and more preferably about 90 to 135 nm. The reconstituted suspension contains 0 to 1.8% (w / v), preferably 0.45% to 1.8% (w / v), more preferably 0.9% to 1.8% (w / v), and even more preferably 0.9% sodium chloride. In some embodiments, the reconstituted suspension contains 2 to 10 mg / mL, preferably 2 to 8 mg / mL of docetaxel.

[0066] The method for preparing the docetaxel albumin composition described in this application is: (1) Dissolve docetaxel in an organic solvent, and after dissolution, obtain an organic phase solution, (2) Taking a human serum albumin solution, adding acid to adjust the pH, obtaining an acid-denatured albumin aqueous solution, and taking another salt solution, (3) A step of mixing the organic phase solution, aqueous phase solution and salt solution to load the drug and obtain a drug loading solution, (4) Dialysis to obtain a suspension after dialysis, and the process includes these steps.

[0067] Here, step (2) optionally includes diluting the human serum albumin solution with sterile water for injection to obtain an albumin diluent, before adding acid to adjust the pH value.

[0068] Here, step (2) optionally includes an incubation step after adjusting the pH value by adding acid.

[0069] Here, step (3) optionally includes a cooling step after mixing and loading the drugs.

[0070] Here, step (4) optionally includes, before dialysis, the step of concentrating the drug loading solution obtained in step (3) to obtain a concentrate.

[0071] Here, step (4) optionally includes a concentration or dilution step to adjust the docetaxel concentration in the post-dialysis suspension after dialysis.

[0072] Here, after step (4), optionally, step (5) of sterilization and filtration is included.

[0073] Here, after step (5), the procedure optionally includes a freeze-drying step (6).

[0074] Here, the docetaxel described in step (1) is in any form, preferably anhydrous docetaxel, docetaxel hemihydrate, or docetaxel trihydrate. When calculated using anhydrous docetaxel, the mass ratio of docetaxel to human serum albumin is 1:(2.0~10.0), preferably 1:(3.0~7.0), more preferably 1:(4.0~6.0), and most preferably about 1:4.5).

[0075] Here, the organic solvent described in step (1) is selected from water-miscible solvents such as ethanol, methanol, acetone, and DMSO, and is preferably ethanol. Based on calculations using anhydrous docetaxel, the above organic phase solution contains 45 to 90 mg / mL, preferably 45 to 70 mg / mL of docetaxel.

[0076] Here, the sodium octanoate content in the human serum albumin solution described in step (2) is 0.12 mmol / g protein or less, preferably 0.08 mmol / g protein or less, and preferably 0.045 to 0.08 mmol / g protein.

[0077] Here, in step (2), the albumin diluent contains 6 to 25 mg / mL, preferably 10 to 20 mg / mL, more preferably 12 to 18 mg / mL, and even more preferably about 15 mg / mL of albumin.

[0078] Here, in step (2), the acid is selected from acidic amino acids or acidic polypeptides, organic acids, and inorganic acids. The acidic amino acids or acidic polypeptides include, but are not limited to, cysteine ​​hydrochloride and glutathione; the organic acids include, but are not limited to, citric acid and tartaric acid; and the inorganic acids include, but are not limited to, hydrochloric acid and sulfuric acid. The acid is preferably cysteine ​​hydrochloride, glutathione, hydrochloric acid, and more preferably cysteine ​​hydrochloride. The pH value is preferably 3.5 to 5.5, more preferably 3.5 to 5.0, even more preferably 3.8 to 4.7, also preferably 4.0 to 4.5, and most preferably about 4.1.

[0079] Here, in step (2), the incubation refers to adding acid to adjust the pH value, then raising the temperature to 35°C to 42°C, preferably 38°C to 42°C, and incubating for 30 minutes or more, preferably 30 to 60 minutes.

[0080] Here, the salt solution described in step (2) is selected from aqueous solutions of sodium chloride, potassium chloride, sodium sulfate, or magnesium sulfate, preferably an aqueous solution of sodium chloride, and the concentration of the salt solution is 2% or more, preferably 2% to 35%, and more preferably 10% to 20%.

[0081] In step (3), the conditions for drug loading are to raise the temperature of the organic phase solution and the aqueous phase solution to 35°C to 42°C, preferably 38°C to 42°C, mix the three, and then load the drug.

[0082] Here, the temperature reduction described in step (3) refers to reducing the temperature to a temperature lower than room temperature, preferably 0 to 20°C, more preferably 7 to 15°C.

[0083] Here, the concentrate described in step (4) contains docetaxel in an amount of about 4 to 10 mg / mL, preferably 6 to 10 mg / mL, more preferably 7 to 9 mg / mL, and even more preferably 8 mg / mL.

[0084] Here, in step (4), excess small molecule compounds are removed by dialysis, and the type and amount of dialysate and the fractional molecular weight of the dialysate membrane are not particularly limited and can be selected based on the ordinary technical knowledge or experience of those skilled in the art. To facilitate further formulation and clinical use, the dialysate is preferably an aqueous solution of a clinically acceptable osmotic regulator, such as an aqueous solution of sodium chloride, glucose, phosphate, or citrate. In some embodiments, in step (4), dialysis is performed using a sodium chloride solution as the dialysate. The fractional molecular weight of the dialysate membrane is 10 to 50 kDa, preferably 10 to 30 kDa, more preferably 10 kDa or 30 kDa. The volume of the dialysate is three times or more the volume of the drug loading solution or concentrate, preferably 3 to 10 times, more preferably 3 to 6 times. The concentration of the sodium chloride solution is 1.8% (w / v) or less, preferably 0.45% to 1.8% (w / v), more preferably 0.9% to 1.8% (w / v).

[0085] In some embodiments, the human serum albumin solution used in step (2) needs to be pre-adjusted in terms of the sodium octanoate content it contains. Those skilled in the art can adjust the sodium octanoate content in the human serum albumin solution by selecting an appropriate method, including but not limited to dialysis, based on ordinary technical knowledge or experience.

[0086] In some embodiments, methods for adjusting the sodium octanoate content in a human serum albumin solution include taking a commercially available human serum albumin solution, diluting it with sterile water for injection or physiological saline to obtain a diluted albumin solution, or dialyzing the diluted albumin solution with sterile water for injection or physiological saline as the dialysate to partially remove sodium octanoate and obtain a human serum albumin solution with a low sodium octanoate content.

[0087] In some embodiments, the dilution factor of the human serum albumin solution is 4 times or more, preferably 4 to 7 times. In the above dialysis, the molecular weight cutoff of the dialysis membrane is 10 to 50 kDa, preferably 10 to 30 kDa, more preferably 10 kDa or 30 kDa. The volume of the dialysate can be determined by a standard test based on the required sodium octanoate content for those skilled in the art. Preferably, the volume of the dialysate is about 3 times or more the volume of the diluted albumin solution, preferably 3 to 10 times, more preferably 3 to 6 times.

[0088] The sodium octanoate content in the human serum albumin solution with a low sodium octanoate content described above is less than 0.16 mmol / g protein, preferably less than 0.12 mmol / g protein, more preferably less than 0.10 mmol / g protein, and even more preferably less than 0.08 mmol / g protein. The human serum albumin solution with a low sodium octanoate content described above may be used directly in the preparation of the docetaxel albumin nanoparticle composition described in this application, or it may be mixed with other human serum albumin solutions having a sodium octanoate content in a specific ratio to obtain the desired content before being used in the preparation of the docetaxel albumin nanoparticle composition.

[0089] In the above preparation method, the dialysis step removes excess small molecule compounds from the drug loading solution or concentrate. For example, in some embodiments, step (2) involves adjusting the pH with an acidic amino acid or acidic polypeptide to prepare acid-denatured albumin, and the dialysis step essentially removes excess acidic amino acids or acidic polypeptides, so that the composition contains almost no free acidic amino acids or acidic polypeptides. The phrase "containing almost no free acidic amino acids or acidic polypeptides" means that the content of free acidic amino acids or acidic polypeptides in the composition is less than 0.25% (w / w) of docetaxel. For example, in some embodiments, the pH involves adjusting the pH with cysteine ​​hydrochloride or glutathione to prepare acid-denatured albumin, and the content of free cysteine ​​or glutathione in the composition is less than 0.25% (w / w) of docetaxel.

[0090] The relevant sequences of the present invention are as follows:

[0091] TIFF2026517915000001.tif234170TIFF2026517915000002.tif195170

[0092] It should be understood that, within the scope of this application, each of the technical features described above and each of the technical features specifically described below (for example, in the examples) can be combined with each other to form a novel or preferred technical solution. Due to space limitations, the explanation is omitted here. [Brief explanation of the drawing]

[0093] [Figure 1] This is a schematic diagram of the KN026 configuration. [Modes for carrying out the invention]

[0094] The following embodiments are provided to better illustrate the content of the present invention, but are not limited to the embodiments described herein. Those skilled in the art will make non-essential improvements and adjustments to the embodiments based on the above description of the invention, but these will still fall within the scope of the claims of the present invention.

[0095] Unless otherwise specified, the singular forms “one,” “one kind,” and “the said” as used herein also include the plural forms. For example, the term “one cell” includes multiple cells and mixtures thereof.

[0096] As used herein, the terms “about” or “approximately” refer to the general range of error for each value that is readily apparent to those skilled in the art. The use of values ​​or parameters related to “about” or “approximately” herein includes (and describes) embodiments (or examples) relating to the value or parameter itself.

[0097] As used herein, the terms “contains” or “includes” mean including, but not excluding, any other elements, integers, or steps described above. In the present invention, when the terms “contains” or “includes” are used, unless otherwise specified, it also covers the elements, integers, or steps referred to. For example, when referring to an antibody variable region that “contains” a specific sequence, it is also intended to cover the antibody variable region consisting of that specific sequence.

[0098] As used herein, the term “cancer” refers to a proliferative disorder caused by or characterized by cell proliferation, in which the cells have lost their sensitivity to normal growth controls. The term “cancer” includes tumors and any other proliferative disorders. Cancers of the same tissue type originate from the same tissue and can be divided into different subtypes based on their biological characteristics.

[0099] As used herein, the term “treatment” refers to a clinical intervention to alter the natural course of an individual or cell being treated in a clinicopathological process. The ideal effects of treatment include slowing or reducing the rate of disease progression, improving or alleviating the disease state, and mitigating or improving the prognosis. For example, a patient is “treated” if one or more symptoms associated with cancer are alleviated or disappear, if cancer cell proliferation is reduced (or cancer cells disappear), if symptoms of the disease are reduced, if the quality of life of the patient is improved, if the dosage of other drugs required to treat the disease is reduced, if disease progression is delayed and / or if the patient’s survival is extended.

[0100] As used herein, the term “antibody” includes monoclonal antibodies (including full-length antibodies having an immunoglobulin Fc region), antibodies with multiepitope specificity, multispecific antibodies (e.g., bispecific antibodies), dimers and single-chain antibodies, and antibody fragments (antigen-binding fragments, e.g., Fab, F(ab')2 and Fv).

[0101] As used herein, the term "CDR (Complementarity-Determining Region)" refers to an amino acid sequence that limits both the binding affinity and specificity of the antibody's Fv region. This is because antibody specificity lies in the structural complementarity between the antibody-binding site and the antigenic determinant. The antibody-binding site is composed primarily of residues derived from the so-called hypervariable region or complementarity-determining region (CDR). The light chain (L) and heavy chain (H) of an antibody each have three CDRs, and the antigen-binding site of a typical antibody contains six CDRs, including groups of CDRs from both the heavy chain variable region and the light chain variable region.

[0102] As used herein, the term “heavy chain variable region (VH)” refers to the amino-terminal variable region domain of an immunoglobulin heavy chain.

[0103] As used herein, the term “light chain variable region (VL)” refers to the amino-terminal variable region domain of an immunoglobulin light chain.

[0104] As used herein, the term “production unit” refers to the smallest fractional packaging of each drug formulation containing an appropriate amount of the active compound. For example, in capsules, a unit formulation refers to one capsule; in tablets, a unit formulation refers to one tablet; in injections, a unit formulation refers to one vial of injection; and other types of formulations are similarly defined.

[0105] As used herein, the term "objective response rate (ORR)" is defined as the proportion of participants who achieve a complete response (CR) or a partial response (PR) during the study period, and adopts the RECIST 1.1 criteria.

[0106] As used herein, the term “partial response” or “PR” refers to a reduction of at least 30% in the sum of the diameters (SOD) of target lesions compared to baseline, with no progression of non-target lesions and no development of new lesions, adopting the RECIST 1.1 criteria.

[0107] As used herein, the term “Complete response” or “CR” refers to the disappearance of all non-lymph node target and non-target lesions and the absence of new lesions, adopting the RECIST 1.1 criteria.

[0108] As used herein, the term “therapeutic dose” means a quantity that produces a therapeutic effect on a subject, for example, in a subject to which the symptoms or condition of the disease are alleviated, reduced or resolved, or the progression of the symptoms or condition of the disease is delayed or suppressed, compared to a subject to which the dose is not administered.

[0109] As used herein, the term “unacceptable” means that a subject has previously received anticancer treatment, but adverse events (e.g., peripheral neuropathy, skin toxicity, hyperglycemia, gastrointestinal symptoms) occurred during the first two cycles, and treatment was discontinued because the symptoms did not improve despite following dose adjustment guidelines, and there was no evidence of disease progression.

[0110] As used herein, the term "CnDm" refers to day m of the nth administration cycle, for example, C1D1 refers to the time of the first administration and C4D1 refers to day 1 of the fourth administration cycle.

[0111] As used herein, the term “recurrence” refers to the recurrence of cancer (e.g., breast cancer) in which new lesions appear after cancer (e.g., breast cancer) has previously received curative treatment.

[0112] As used herein, the term "metastasis" refers to the spread of a tumor lesion beyond the ipsilateral supraclavicular lymph nodes, ipsilateral internal mammary lymph nodes, ipsilateral axillary lymph nodes, ipsilateral chest wall, skin, etc., and is known as distant metastasis.

[0113] In some embodiments, the anti-HER2 antibody comprises two different heavy chains (H) and two common light chains (L) with identical sequences. Each common light chain has three CDRs, each named CDR-L1, CDR-L2, and CDR-L3. In some embodiments, the common light chain includes CDR-L1 shown in SEQ ID NO: 1, CDR-L2 shown in SEQ ID NO: 2, and CDR-L3 shown in SEQ ID NO: 3. The heavy chain includes heavy chain I, which targets HER2 protein domain II, and heavy chain II, which targets HER2 protein domain IV. Heavy chain I has three CDRs, each named CDR-H1-1, CDR-H1-2, and CDR-H1-3. In some embodiments, heavy chain I includes CDR-H1-1 shown in SEQ ID NO: 6, CDR-H1-2 shown in SEQ ID NO: 7, and CDR-H1-3 shown in SEQ ID NO: 8. The heavy chain II described above has three CDRs named CDR-H2-1, CDR-H2-2, and CDR-H2-3, respectively. In some embodiments, the heavy chain II includes CDR-H2-1 shown in SEQ ID NO: 11, CDR-H2-2 shown in SEQ ID NO: 12, and CDR-H2-3 shown in SEQ ID NO: 13.

[0114] In this application, heavy chain I and heavy chain II are simply distinguished as two heavy chains targeting different epitopes, and do not indicate any order or positional significance.

[0115] The docetaxel albumin composition described herein is also called docetaxel (albumin-bound). In some embodiments, the docetaxel albumin composition is a docetaxel albumin nanoparticle composition, which is prepared using conventional methods for preparing albumin nanoparticle drugs in the art. Human serum albumin, being an endogenous human substance, possesses excellent biocompatibility and, as a natural carrier for hydrophobic drugs, increases the solubility of poorly soluble drugs and enhances therapeutic effects by utilizing albumin's unique "gp60-caveolin-SPARC" channel to concentrate drugs in tumor regions.

[0116] In some embodiments, the docetaxel albumin compositions described herein are prepared by reference to the methods disclosed in WO2022184164A1, all of which are incorporated herein by reference.

[0117] In some embodiments, the anti-Her2 bispecific antibody disclosed herein is KN026. The sequences of the two common light chains of KN026 are both the amino acid sequences shown in SEQ ID NO: 5, the sequence of heavy chain I is the amino acid sequence shown in SEQ ID NO: 10, and the sequence of heavy chain II is the amino acid sequence shown in SEQ ID NO: 15. The structure of KN026 is shown in Figure 1. The KN026 antibody can be prepared using conventional articulation methods.

[0118] Fab1 (heavy chain I) of the KN026 antibody protein specifically recognizes domain II of the human HER2 protein and can block signaling by heterodimerization by HER2 and other ERBB family members, while Fab2 (heavy chain II) inhibits HER2-related signaling pathways through domain IV, which targets the human HER2 protein. The synergistic action of these two inhibitory mechanisms allows the KN026 bispecific antibody to more effectively inhibit the proliferation of HER2-positive tumor cells. Furthermore, the CH3 domains of the two heavy chains of KN026 contain a series of asymmetric amino acid mutations to increase the yield of heterodimeric antibody proteins while still maintaining complete Fc function, giving KN026 clear antibody-dependent cell-mediated cytotoxicity. In vitro antitumor cell proliferation experiments demonstrate that the antibody inhibits tumor cell proliferation in a dose-dependent manner. In some embodiments, the preparation of KN026 and related formulations disclosed herein can be obtained by methods conventional in the art; for example, the formulation may be a liquid formulation. For example, the formulation may be prepared in an aqueous carrier. For example, a stabilizer may be added in an amount below an undesirable viscosity or viscosity unsuitable for intravenous administration. For example, the liquid formulation may further contain one or more of the following: buffers, surfactants, and preservatives.

[0119] Preparation example The docetaxel (albumin-bound) for injection used in the examples is produced and supplied by Shiyaku Group and prepared by referring to the method described in Example 1 Formulation 1-1 of Patent WO2022184164A1. Specifically, it is as follows:

[0120] (1) 8 g of anhydrous docetaxel was weighed and dissolved in 120 mL of ethanol to obtain an organic phase solution after dissolution.

[0121] (2) A human serum albumin solution containing 36 g of albumin (containing 0.08 mmol / g of sodium octanoate protein) was taken, diluted with sterile water for injection in a solution containing 15 mg / mL of albumin, an appropriate amount of cysteine ​​hydrochloride was added to adjust the pH to 4.1, and incubated at 42°C for 30 minutes to obtain an acid-denatured albumin aqueous phase solution. 18 g of sodium chloride was taken and prepared as a 20% salt solution with sterile water for injection.

[0122] (3) The organic phase solution and the aqueous phase solution were heated to 42°C, the organic phase, aqueous phase and salt solution were mixed, and the drug was added. The resulting material was then cooled to 15°C in an ice bath to obtain the drug-loaded solution.

[0123] (4) The drug loading solution was concentrated until the docetaxel concentration reached approximately 8 mg / mL to obtain the concentrate. An isotonic sodium chloride solution (0.9%, w / v) was used as the dialysate, and the concentrate was dialyzed five times. The molecular weight cutoff of the dialysate membrane was 30 kDa. A suspension was obtained after dialysate, and then an appropriate amount of dialysate was added to adjust the concentration of the suspension until the docetaxel concentration reached 4 mg / mL.

[0124] (5) The suspension was sterilized using a 0.45 μm + 0.2 μm film sheet and filtered to obtain a suspension before freeze-drying.

[0125] (6) The suspension obtained in step (5) before freeze-drying was taken and freeze-dried to obtain freeze-dried powder.

[0126] KN026 contains heavy chain I, whose amino acid sequence is shown in SEQ ID NO: 10; heavy chain II, whose amino acid sequence is shown in SEQ ID NO: 15; and a common light chain, whose amino acid sequence is shown in SEQ ID NO: 5.

[0127] A vector containing nucleic acid sequences capable of encoding the above-mentioned heavy chain I, heavy chain II, and common light chain was transferred to cells, expressed, and purified to obtain KN026. The specific preparation process can be found in patent application WO2016110267.

[0128] Example 1: Clinical Study This is a randomized, controlled, open-label, multicenter, priority Phase III clinical study to compare the efficacy and safety of KN026 in combination with injectable docetaxel (albumin-bound) and trastuzumab in combination with pertuzumab and docetaxel as first-line treatments for HER2-positive recurrent or metastatic breast cancer.

[0129] 1. Research method 1.1. Information on test drugs and dosage plan: (1) Information on test chemicals

[0130] All experimental drugs were provided by TIFF2026517915000003.tif162170.

[0131] (2) Dosage plan for the test group [1]KN026 The drug was administered once every three weeks, on the first day of each cycle, at a dose of 30 mg / kg. The initial infusion was administered intravenously for 90 min (±15 min), and if there was no infusion response, the infusion time for subsequent cycles could be adjusted to 60 min (±15 min). Each vial contained 325 mg (13 mL) of KN026 antibody. The required drug volume was calculated according to body weight, the corresponding volume of drug was extracted, diluted in 250 mL of physiological saline, and administered intravenously.

[0132] [2] Docetaxel for injection (albumin-bound) The drug is administered once every three weeks, on the first day of each cycle, at a dose of 100 mg / m². 2 Therefore, intravenous infusion was administered for 60 minutes (±10 minutes).

[0133] Each vial contained 80 mg of docetaxel (albumin-bound) powder for injection. This was dispersed and dissolved in 20 mL of sterile water for injection to form a 4 mg / mL suspension. The corresponding administration volume was calculated based on the subject's body surface area and injected into an infusion bag. Simultaneously, the same volume of physiological saline was taken and diluted to prepare a 2 mg / mL solution, which was then administered by intravenous infusion.

[0134] The administration order in the test group was KN026 followed by docetaxel (albumin-bound) for injection, and it was recommended to observe the patients for at least 30 minutes between the two drug administrations.

[0135] To prevent severe hematological toxicity, granulocyte colony-stimulating factor (G-CSF) could be prophylactically administered 24-72 hours after the completion of infusion of injectable docetaxel (albumin-bound).

[0136] (3) Dosage plan for the control group: [1] Pertuzumab injection The drug was administered once every three weeks, on day 1 of each cycle. The initial dose was 840 mg, administered intravenously for 60 min (±10 min). Subsequently, the dose for each cycle was 420 mg, with an infusion time of 30-60 min (±10 min). Note: If delayed administration resulted in a ≥6-week interval between two consecutive infusions, pertuzumab was re-administered as a loading dose of 840 mg, followed by continued administration of 420 mg every three weeks.

[0137] Each vial contained 14 mL of concentrated solution with a concentration of 30 mg / mL. The corresponding dose volume was calculated and administered intravenously in 250 mL of physiological saline. Note: Pertuzumab should not be diluted with 5% glucose solution due to its unstable chemical and physical properties in 5% glucose solution.

[0138] [2] Trastuzumab for injection The drug was administered once every three weeks, on day 1 of each cycle. The initial dose was 8 mg / kg, administered intravenously for 90 min (±10 min), followed by a dose of 6 mg / kg for 30-90 min (±10 min) in each subsequent cycle. If delayed administration resulted in an interval of ≥6 weeks between two consecutive infusions, trastuzumab was re-administered as a loading dose of 8 mg / kg, followed by continued administration of 6 mg / kg every three weeks.

[0139] The appropriate dosage volume was calculated based on the subject's weight, and intravenous infusion was administered in addition to 250 mL of physiological saline.

[0140] [3] Docetaxel injection It is administered once every three weeks, on the first day of each cycle, at a dose of 75 mg / m². 2 Therefore, the infusion was administered for more than 60 minutes each time.

[0141] Prior to administering docetaxel injection, pretreatment was required according to the instructions for use [oral administration of a glucocorticoid, such as dexamethasone, the day before docetaxel injection, at a dose of 16 mg per day (e.g., 8 mg twice daily for 3 days)] or standard clinical practice.

[0142] The control group received pertuzumab, trastuzumab, and docetaxel in that order, and it was recommended to observe the patient for at least 30 minutes between the administration of the two drugs.

[0143] (4) Dosage adjustment In this study, dose reduction was not permitted for KN026, trastuzumab, and pertuzumab. Dose reduction was permitted if adverse events related to treatment occurred with docetaxel injectable (albumin-bound) or docetaxel injection. For docetaxel injectable (albumin-bound), a maximum of two dose downregulations were permitted; if a third dose reduction was necessary, permanent discontinuation was required. For docetaxel injection, only one dose downregulation was permitted; if a second dose reduction was necessary, permanent discontinuation was required.

[0144] [Table 2]

[0145] 1.2. Selection and Exclusion Criteria Selection Criteria To participate in this study, participants must meet all of the following selection criteria.

[0146] 1) I voluntarily participated in this study and signed the Informed Consent Form (ICF).

[0147] 2) Age ≥ 18 years old.

[0148] 3) Histologically and / or cytologically confirmed diagnosis of recurrent or metastatic breast cancer.

[0149] 4) The latest tumor tissue sample was confirmed to be HER2-positive (IHC3+, or IHC2+ and ISH-positive) by detection in the central laboratory.

[0150] 5) Participation was considered for patients who had not previously received systemic chemotherapy and / or HER2-targeted therapy for recurrent or metastatic breast cancer (participation was permitted for patients who had received one endocrine therapy program) and who had relapsed at least 12 months after completion of (new) adjuvant chemotherapy or HER2-targeted therapy.

[0151] 6) The physical fitness score for the East Coast Cancer Clinical Group (ECOG) was between 0 and 1 point.

[0152] 7) At least one measurable or evaluable lesion (RECIST 1.1) was present.

[0153] 8) The patient had adequate organ and bone marrow function (no blood transfusions within 14 days prior to detection, and no correction with hematopoietic stimulating agents). a) Neutrophil count ≥ 1.5 × 10 9 / L. b) Platelet count ≧100×10 9 / L. c) Hemoglobin ≥ 90 g / L. d) Liver function: Total bilirubin (TBIL) ≤ 1.0 × upper limit of normal (ULN), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ≤ 2.5 × ULN (target for liver metastasis ≤ 5 × ULN). e) Renal function: Creatinine clearance ≥ 30 mL / min (calculated using the Cockcroft-Gault formula). f) Blood coagulation function: International normalized ratio (INR) ≤ 1.5 and activated partial thromboplastin time (APTT) ≤ 1.5 × ULN (For subjects receiving anticoagulation therapy, the researchers determined that both INR and APTT were within a safe and effective therapeutic range). g) Left ventricular ejection fraction (LVEF)>50%.

[0154] 9) Expected survival time ≥ 3 months.

[0155] 10) Women of childbearing age were required to undergo a blood pregnancy test within 7 days prior to randomization, with a negative result, and to consent to reliable and effective contraception within 3 months (docetaxel injection and docetaxel for injection (albumin-bound)) or 7 months (trastuzumab, pertuzumab, and KN026) after the study treatment period and the final study treatment (as per the maximum time limit). Men whose partners were women of childbearing age were required to consent to reliable and effective contraception within 3 months (docetaxel injection and docetaxel for injection (albumin-bound)) or 7 months (trastuzumab, pertuzumab, and KN026) after the study treatment period and the final study treatment (as per the maximum time limit).

[0156] Exclusion criteria Individuals who meet any one of the following criteria are not permitted to participate in this study.

[0157] 1) Individuals who have contraindications to trastuzumab, pertuzumab, and docetaxel, or who have been determined by researchers to be unsuitable for receiving KN026 or injectable docetaxel (albumin-bound).

[0158] 2) Individuals with ALT and / or AST > 1.5 × ULN, and simultaneously with alkaline phosphatase (ALP) > 2.5 × ULN.

[0159] 3) Participants who have peripheral neuropathy of grade ≥ 3 at the time of randomization.

[0160] 4) Patients whose toxicity from previous antitumor treatments has not recovered to a level of Grade 1 or lower according to the Common Terminology Criteria for Adverse Events (CTCAE 5.0) (excluding toxicities that researchers have determined to pose no safety risk, such as alopecia, peripheral neurotoxicity, and simple laboratory tests, which should be relaxed to Grade 2 or lower).

[0161] 5) Patients who, during the course of previous anti-HER2 drug treatment, experienced a decrease in LVEF to <50%, developed symptomatic congestive heart failure (CHF), or experienced related toxicity leading to permanent discontinuation of treatment.

[0162] 6) Anyone who is aware of an allergy to and / or a contraindication to glucocorticoids (including, but not limited to, active gastrointestinal ulcers, severe hypertension, severe hypokalemia, glaucoma, etc.).

[0163] 7) Patients who have previously received treatment with anthracycline drugs and whose cumulative dose has reached the following levels: doxorubicin or liposomal doxorubicin > 360 mg / m² 2 Epirubicin > 720 mg / m² 2 Mitoxantrone > 120 mg / m² 2 Other anthracycline drugs > 360 mg / m² 2 When using doxorubicin equivalent to this, or one or more anthracycline drugs, the cumulative dose is 360 mg / m². 2 The amount of doxorubicin equivalent to this must not exceed the appropriate level.

[0164] 8) Patients who have used a potent CYP3A4 inhibitor within 14 days prior to randomization.

[0165] 9) Any person with a history of contraindications or hypersensitivity reactions to any research drug component or any known excipient.

[0166] 10) In cases of untreated or unstable brain parenchymal metastases, spinal cord metastases or compression, or carcinomatous meningitis, participation was considered for patients who had received local brain treatment, whose symptoms were stable, who had been stable for at least 4 weeks prior to randomization on imaging studies, who had no evidence of cerebral edema, and who did not require glucocorticoid therapy.

[0167] 11) Individuals with active malignant tumors within three years prior to randomization, excluding breast cancer studied in this study and any locally curable tumors that have received curative treatment (e.g., excised basal cell or squamous cell skin cancer, superficial bladder cancer, cervical cancer or in-situ breast cancer, early thyroid cancer, etc.).

[0168] 12) Persons with uncontrolled or significant cardiovascular disease, including but not limited to the following: a) Within 6 months prior to randomization, the patient had experienced congestive heart failure of New York Heart Association (NYHA) grade II or higher, unstable angina, myocardial infarction, or an arrhythmia causing hemodynamic instability. b) Primary cardiomyopathy (e.g., dilated cardiomyopathy, hypertrophic cardiomyopathy, arrhythmic right ventricular cardiomyopathy, restrictive cardiomyopathy, atypical cardiomyopathy), c) A history of clinically significant QT interval prolongation, or a screening period QTcF (calculated using the Fridericia formula) > 450 ms. d) Within 6 months prior to randomization, an arterial / venous thromboembolic event other than cerebrovascular events (including transient ischemic stroke, cerebral hemorrhage, and cerebral infarction), deep vein thrombosis, and pulmonary embolism occurred. e) Uncontrolled hypertension (systolic blood pressure > 160 mmHg and / or diastolic blood pressure > 100 mmHg).

[0169] 13) Individuals who have a severe chronic or active infection (including Mycobacterium tuberculosis) requiring systemic antimicrobial, antifungal, or antiviral treatment within 14 days prior to randomization.

[0170] 14) Active hepatitis B (positive for hepatitis B surface antigen and HBV-DNA ≥ 2 × 10⁻¹⁰) 3 Individuals with hepatitis C (positive for hepatitis C antibody and HCV-RNA quantitative detection result > lower limit of detection). Note: Lower limit of detection <HBV DNA<2×10 3 For hepatitis B virus carriers with a dose of IU / mL, participation was considered for those willing to receive entecavir or other antiviral treatment during the study period based on clinical judgment.

[0171] 15) Individuals with a history of human immunodeficiency virus (HIV) infection.

[0172] 16) Individuals currently participating in another clinical study (excluding follow-up periods for non-interventional or interventional clinical studies), or whose time since the end of their previous clinical study (last dose) is less than four weeks at the time of randomization.

[0173] 17) Individuals who have received antitumor treatment such as radiotherapy, targeted therapy, immunotherapy, or other research drugs within 28 days prior to randomization, or who have used herbal medicines with antitumor indications within 14 days prior to randomization.

[0174] 18) Patients who, within 7 days prior to randomization, have uncontrolled serosal fluid accumulation (e.g., pleural effusion, ascites, pericardial effusion) requiring frequent drainage or medical intervention, or who require additional intervention (excluding cytological detection of exudate) within 2 weeks after intervention.

[0175] 19) Individuals who have undergone major organ surgery (excluding fine-needle aspiration biopsy) within 28 days prior to randomization.

[0176] 20) Pregnant or lactating women.

[0177] 21) Any person who has other circumstances that would prevent him from participating in the research program, or that would not be in his best interest to participate in the research, or that could affect the research results, such as a history of mental illness, drug abuse or substance abuse, or any other clinically significant illness or condition.

[0178] 1.3. Evaluation Metrics 1.3.1. Evaluation of treatment effectiveness RECIST 1.1 was used to evaluate the radiographic efficacy of the treatment. The evaluation of treatment efficacy included the following:

[0179] ORR: The proportion of CR and PR cases in the total study area (assessed by BIRC and researchers). (PR: Compared to the baseline total diameter of target lesions, the total diameter of target lesions decreased by ≥30%, non-target lesions did not progress, and no new lesions developed.)

[0180] CR: This refers to the complete disappearance of all target and non-target lesions in non-lymph nodes, and the absence of new lesion development. Throughout the entire research process, periodic safety assessments were conducted, including physical examinations, vital signs, weight, 12-lead electrocardiograms, ultrasound-magnetocardiograms, and laboratory examinations. The severity of adverse events was recorded using the National Cancer Institute Common Terminology Criteria for Adverse Events, version 5.0 (NCI-CTCAE 5.0).

[0181] 2. Research results (1) The results of the evaluation of the treatment effect are as follows: By February 2024, a total of 267 subjects had been enrolled, of which 170 underwent post-baseline treatment efficacy evaluation. These included 85 subjects in the trial group and 85 in the control group (trastuzumab + pertuzumab + docetaxel). In the trial group, 67 achieved a partial response (PR) with an overall response rate (ORR) of 78.8%, while in the control group, 56 achieved a PR with an ORR of 65.9%. Both the PR and ORR values ​​in the trial group were significantly higher than those in the control group, demonstrating a trend towards superior antitumor therapeutic efficacy in the trial group compared to the control group.

[0182] (2) The safety evaluation results are as follows: To date, no new adverse reaction signals have been detected in the test group of this invention. Treatment of HER2-positive recurrent metastatic breast cancer with a combination of anti-HER2 antibody (KN026) and docetaxel albumin composition exhibits superior antitumor efficacy compared to conventional standard treatment plans, has better overall safety, and is more convenient to administer. This provides such patients with a first-line treatment option that offers superior efficacy, better safety, and greater convenience.

Claims

1. The use of an anti-HER2 antibody and a second therapeutic agent in the preparation of a drug for treating cancer, wherein the second therapeutic agent is a docetaxel albumin composition. use.

2. The use of an anti-HER2 antibody in the preparation of a drug to enhance the therapeutic effect of a second therapeutic agent for treating cancer, wherein the second therapeutic agent is a docetaxel albumin composition. use.

3. A method for treating cancer with an anti-HER2 antibody and a second therapeutic agent, wherein the second therapeutic agent is a docetaxel albumin composition. method.

4. A pharmaceutical composition, reagent kit, or drug kit for treating cancer, comprising an anti-HER2 antibody and a second therapeutic agent, wherein the second therapeutic agent is a docetaxel albumin composition. Pharmaceutical compositions, reagent kits, or drug kits.

5. The anti-HER2 antibody is an anti-HER2 bispecific antibody, preferably a bispecific antibody that targets different epitopes of the human HER2 protein, and more preferably, the different epitopes of the HER2 protein are domain II and domain IV. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 4.

6. The anti-HER2 antibody comprises two common light chains having the same sequence, preferably comprising CDR-L1 shown in SEQ ID NO: 1, CDR-L2 shown in SEQ ID NO: 2, and CDR-L3 shown in SEQ ID NO: 3, more preferably comprising the light chain variable region shown in SEQ ID NO: 4, and even more preferably comprising the amino acid sequence shown in SEQ ID NO:

5. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 5.

7. The anti-HER2 antibody comprises two heavy chains with different sequences, preferably a heavy chain I targeting HER2 protein domain II and / or a heavy chain II targeting HER2 protein domain IV. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 6.

8. The heavy chain I comprises CDR-H1-1 shown in SEQ ID NO: 6, CDR-H1-2 shown in SEQ ID NO: 7, and CDR-H1-3 shown in SEQ ID NO: 8, preferably the heavy chain I comprises the heavy chain I variable region shown in SEQ ID NO: 9, more preferably the heavy chain I comprises the amino acid sequence shown in SEQ ID NO: 10 or SEQ ID NO: 16, and / or The heavy chain II comprises CDR-H2-1 shown in SEQ ID NO: 11, CDR-H2-2 shown in SEQ ID NO: 12, and CDR-H2-3 shown in SEQ ID NO: 13, preferably the heavy chain II comprises the heavy chain II variable region shown in SEQ ID NO: 14, and more preferably the heavy chain II comprises the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO:

17. The use, method, pharmaceutical composition, reagent kit, or drug kit according to claim 7.

9. The docetaxel albumin composition comprises docetaxel and acid-modified albumin, preferably the docetaxel is selected from anhydrous docetaxel, and more preferably the weight ratio of the docetaxel to the albumin is 1:(4.0 to 6.0). The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 8.

10. The acid-denatured albumin is obtained by denaturing human serum albumin after adding an acid to adjust the pH to an appropriate value, preferably the acid being cysteine ​​hydrochloride, and preferably the pH value being about 4.

1. The use, method, pharmaceutical composition, reagent kit, or drug kit according to claim 9.

11. The docetaxel albumin composition further comprises an osmotic pressure regulator and sodium octanoate, preferably the osmotic pressure regulator is sodium chloride, more preferably the weight ratio of sodium chloride to docetaxel is about 2.25:1, preferably the sodium octanoate content is less than 0.16 mmol / g protein, and more preferably the sodium octanoate content is less than 0.08 mmol / g protein. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 10.

12. The aforementioned cancers include, but are not limited to, hematological malignancies or solid tumors. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 11.

13. The aforementioned hematological malignancies are lymphomas or leukemias, and include, but are not limited to, myeloma, B-cell lymphoma, mantle cell lymphoma, non-Hodgkin B-cell lymphoma, non-Hodgkin T-cell lymphoma, cutaneous lymphoma, anaplastic large cell lymphoma, multiple myeloma, inactive non-Hodgkin lymphoma, plasmacytoma, chronic lymphocytic leukemia, small lymphocytic lymphoma, and follicular lymphoma. Squamous tumors include, but are not limited to, bladder cancer, brain cancer, breast cancer, cervical cancer, thoracic cancer, endometrial cancer, esophageal squamous cell carcinoma, stomach cancer, head cancer, pancreatic cancer, bile duct cancer, colorectal cancer, eye cancer, head and neck squamous cell carcinoma, urothelial carcinoma, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, laryngeal cancer, and uterine cancer, melanoma, skin cancer, salivary gland cancer, soft tissue sarcoma, and osteosarcoma. The use, method, pharmaceutical composition, reagent kit, or drug kit according to claim 12.

14. The solid tumor is breast cancer, preferably HER2-positive breast cancer, and more preferably HER2-positive recurrent and / or metastatic breast cancer. The use, method, pharmaceutical composition, reagent kit, or drug kit according to claim 12.

15. The anti-HER2 antibody and the second therapeutic agent are contained in the same formulation unit, or they are each contained in separate formulation units. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 14.

16. The docetaxel albumin composition is prepared into a formulation, and each formulation unit contains 50 to 100 mg, preferably 70 to 90 mg, more preferably about 80 mg of docetaxel (calculated using anhydrous docetaxel), and / or the anti-HER2 antibody is prepared into a formulation, and each formulation unit contains the anti-HER2 antibody or its pharmaceutically acceptable form (calculated using anhydrous anti-HER2 antibody) in the range of about 0.01 mg to about 400 mg, preferably 300 to 400 mg, more preferably about 300 mg, about 325 mg, about 350 mg, about 375 mg, or about 400 mg. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 15.

17. The docetaxel albumin composition contains 60 to 125 mg / m². 2 Preferably 60 to 100 mg / m² 2 Comfortably approximately 60 mg / m² 2 , about 75 mg / m 2 Alternatively, approximately 100 mg / m² 2 The anti-HER2 antibody is administered in doses of 0.01 mg / kg to 100 mg / kg, preferably 20 to 40 mg / kg, 20 mg / kg to 30 mg / kg, 25 mg / kg to 35 mg / kg, and more preferably about 30 mg / kg. The use, method, pharmaceutical composition, reagent kit, or drug kit according to any one of claims 1 to 16.

Images