TW202432185A

TW202432185A - Ligand-cytotoxicity drug conjugates and pharmaceutical uses thereof

Info

Publication number: TW202432185A
Application number: TW112141379A
Authority: TW
Inventors: 林宗逸; 佩芝黃; 志鴻羅
Original assignee: 美商翰森生物有限責任公司; 大陸商上海翰森生物醫藥科技有限公司; 大陸商常州恆邦藥業有限公司
Priority date: 2022-10-28
Filing date: 2023-10-27
Publication date: 2024-08-16
Also published as: WO2024088388A1

Abstract

Disclosed are the antibody and the conjugate of the antibody and small molecule medicine. The antibody have capacity of binding to human antigen with high affinity, and have better internalization properties. Also disclosed are uses of the ligand-cytotoxicity drug conjugate and pharmaceutical compositions comprising the same in the preparation of drugs for treating cancers.

Description

Ligand-cytotoxic drug conjugate and its drug use

本發明涉及一種新型HER3抗體或其功能片段，其包含工程改造的重鏈和輕鏈。本發明進一步涉及改進的HER3抗體與小分子藥物的綴合物。本發明進一步涉及該抗體和該綴合物在製造用於治療癌症的藥物中的用途。 The present invention relates to a novel HER3 antibody or a functional fragment thereof, which comprises an engineered heavy chain and a light chain. The present invention further relates to a conjugate of an improved HER3 antibody and a small molecule drug. The present invention further relates to the use of the antibody and the conjugate in the manufacture of a drug for treating cancer.

人表皮生長因子受體3(ErbB3，也稱為HER3)是一種酪胺酸激酶受體蛋白，並且屬於酪胺酸激酶受體蛋白的表皮生長因子受體(EGFR)亞家族，該亞家族還包括EGFR(HER1，ErbB1)、HER2(ErbB2，Neu)和HER4(ErbB4)。此外，HER3是一個獨特的HER家族成員，其沒有或幾乎沒有細胞內酪胺酸激酶活性，但仍在腫瘤進展和耐藥性中起作用。 Human epidermal growth factor receptor 3 (ErbB3, also known as HER3) is a tyrosine kinase receptor protein and belongs to the epidermal growth factor receptor (EGFR) subfamily of tyrosine kinase receptor proteins, which also includes EGFR (HER1, ErbB1), HER2 (ErbB2, Neu), and HER4 (ErbB4). In addition, HER3 is a unique HER family member that has no or almost no intracellular tyrosine kinase activity, but still plays a role in tumor progression and drug resistance.

藉由使用單株抗體，針對HER家族成員(如EGFR和HER2)的靶向療法廣泛而通常地用於癌症療法。然而，由於癌症治療的適應性特徵，一些癌症患者在長期治療後產生耐藥性。例如，約70%的患者對曲妥珠單抗(一種抗HER2抗體)耐藥，並且一些患者甚至表現出原發耐藥性。數項研究報告稱，在腫瘤對曲妥珠單抗治療產生耐藥性後，觀察到HER3 的強烈表達，因此HER3成為克服現有障礙的有前景的靶點。先前研究顯示，HER3表達不僅與轉移事件有關，而且還涉及對用其他EGFR靶向療法(例如西妥昔單抗，或激酶抑制劑，如拉帕替尼)、IGFR靶向療法或化學治療劑治療的耐藥性的發生。 Targeted therapies against HER family members, such as EGFR and HER2, are widely and commonly used in cancer therapy by using monoclonal antibodies. However, due to the adaptive nature of cancer treatment, some cancer patients develop resistance after long-term treatment. For example, about 70% of patients are resistant to trastuzumab, an anti-HER2 antibody, and some patients even show primary resistance. Several studies have reported that strong expression of HER3 was observed after tumors developed resistance to trastuzumab treatment, making HER3 a promising target to overcome existing obstacles. Previous studies have shown that HER3 expression is not only associated with metastatic events, but also involved in the development of resistance to treatment with other EGFR-targeted therapies (e.g., cetuximab, or kinase inhibitors such as lapatinib), IGFR-targeted therapies, or chemotherapy.

因為HER3僅具有極小的激酶活性，所以針對HER3的抗體是迄今為止靶向HER3的最受追求的策略。許多針對HER3的單株抗體正在進行臨床前和臨床開發。目前，活性抗體是來自Hummingbird Bioscience的HMBD-001(NCT05057013)；來自ISU Abxis Co.,Ltd.的ISU104(NCT03552406)；和來自上海生物製品研究所的SIBP-03(NCT05203601)。然而，全人IgG2 mAb瑟瑞妥單抗(Seribantumab)(來自Merrimack，MM-121)與紫杉醇或依西美坦(芳香酶抑制劑)組合在HER3+卵巢癌和乳腺癌患者中未達到無進展生存期(PFS)的2期臨床終點(NCT03241810)。此外，在評估其對NSCLC的療效的3期臨床研究中，對來自Daiichi-Sankyo的帕曲妥單抗(Patritumab)(U3-1287/AMG888)與厄洛替尼一起進行了測試，帕曲妥單抗也未能達到療效標準(NCT02134015)。很多HER3靶向治療性抗體已經嘗試在臨床試驗中治療患者，但它們的療效只能被視為適度的(J Exp Clin Cancer Res.2022年10月21日；41(1)：310)。因此，迫切需要併入新策略以改善HER3靶向抗體療法。 Because HER3 has only minimal kinase activity, antibodies against HER3 are by far the most sought-after strategy for targeting HER3. Many monoclonal antibodies against HER3 are in preclinical and clinical development. Currently, the active antibodies are HMBD-001 from Hummingbird Bioscience (NCT05057013); ISU104 from ISU Abxis Co., Ltd. (NCT03552406); and SIBP-03 from Shanghai Institute of Biological Products (NCT05203601). However, the fully human IgG2 mAb seribantumab (from Merrimack, MM-121) did not meet the phase 2 endpoint of progression-free survival (PFS) in patients with HER3+ ovarian and breast cancer in combination with paclitaxel or exemestane (aromatase inhibitors) (NCT03241810). In addition, in a phase 3 clinical study evaluating its efficacy in NSCLC, patritumab (U3-1287/AMG888) from Daiichi-Sankyo was tested together with erlotinib and also failed to meet the efficacy criteria (NCT02134015). Many HER3-targeted therapeutic antibodies have been tried to treat patients in clinical trials, but their efficacy can only be considered modest (J Exp Clin Cancer Res. 2022 Oct 21;41(1):310). Therefore, there is an urgent need to incorporate new strategies to improve HER3-targeted antibody therapy.

儘管首創ADC德帕瑞妥單抗(patritumab deruxtecan)取得了有前景的結果，但調節HER3在癌症患者中功能的複雜機制使得有必要進一步研究針對HER3的新型且優化的治療策略。因此，需要開發調節HER3 活性的新型、有效且安全的產品，從而治療HER3相關疾病；需要併入更多樣化的組分和組合，以開發更先進的抗HER3 ADC。本發明提供了滿足這一未滿足需要的技術方案。 Despite promising results from the first-in-class ADC patritumab deruxtecan, the complex mechanisms regulating HER3 function in cancer patients warrant further investigation of novel and optimized therapeutic strategies targeting HER3. Therefore, there is a need to develop novel, effective, and safe products that modulate HER3 activity to treat HER3-related diseases; more diverse components and combinations need to be incorporated to develop more advanced anti-HER3 ADCs. The present invention provides a technical solution to meet this unmet need.

抗體-藥物綴合物(ADC)的抗癌功效被認為依賴於表達表面抗原的癌細胞攝入它們，因此靶向HER3的單株抗體的內化不足是待解決的緊急問題。所有先前的抗HER3抗體都是基於結合而不是內化而產生的，內化對於ADC至關重要。此外，所有抗HER3抗體均由噬菌體展示技術或傳統小鼠融合瘤技術分離。我們利用全人源化小鼠篩選抗HER3的全人抗體，這與所有先前的抗HER3抗體完全不同。 The anticancer efficacy of antibody-drug conjugates (ADCs) is believed to be dependent on their uptake by cancer cells expressing surface antigens, so the lack of internalization of monoclonal antibodies targeting HER3 is an urgent problem to be solved. All previous anti-HER3 antibodies were generated based on binding rather than internalization, which is crucial for ADCs. In addition, all anti-HER3 antibodies were isolated by phage display technology or traditional mouse hybridoma technology. We screened fully human antibodies against HER3 using fully humanized mice, which is completely different from all previous anti-HER3 antibodies.

本發明要解決的技術問題是開發在癌症中表現出非常強的抗腫瘤效果的先進的抗HER3 ADC。可以藉由以下來增進抗腫瘤效果：抗體的改善的結合親和力、用於強勁內吞作用的內化增加、更高的DAR以及不受NRG1抑制的ADC的一些潛在效果。 The technical problem to be solved by the present invention is to develop an advanced anti-HER3 ADC that exhibits very strong anti-tumor effects in cancer. The anti-tumor effect can be enhanced by the following: improved binding affinity of the antibody, increased internalization for strong endocytosis, higher DAR, and some potential effects of ADCs that are not inhibited by NRG1.

具體地，本發明涵蓋以下方面： Specifically, the present invention covers the following aspects:

本揭露提供了通式(A)的抗體-藥物綴合物或其藥學上可接受的鹽或溶劑化物， The present disclosure provides an antibody-drug conjugate of general formula (A) or a pharmaceutically acceptable salt or solvate thereof,

Ab-(LAb-(L ₂₂ -L-L ₁₁ -D)-D) _yy (A)(A)

其中， in,

D是細胞毒性藥物； D is a cytotoxic drug;

L₁和L₂是連接單元； _L1 and _L2 are connection units;

y是1至20的數，較佳2至10，更佳2至8，更佳2至6或4至8，最佳2、4、6、7、8； y is a number from 1 to 20, preferably from 2 to 10, more preferably from 2 to 8, more preferably from 2 to 6 or 4 to 8, and most preferably from 2, 4, 6, 7, 8;

Ab是抗HER3抗體或其抗原結合片段，其包含：包含HCDR1、HCDR2和HCDR3區的抗體重鏈可變區，和包含LCDR1、LCDR2和LCDR3區的抗體輕鏈可變區，其中：a)HCDR1如SEQ ID NO：01、SEQ ID NO：02、SEQ ID NO：03、SEQ ID NO：04、SEQ ID NO：05、SEQ ID NO：06、SEQ ID NO：07、SEQ ID NO：08、SEQ ID NO：09、SEQ ID NO：10、SEQ ID NO：11、SEQ ID NO：12、SEQ ID NO：13、SEQ ID NO：14、SEQ ID NO：15、SEQ ID NO：16或SEQ ID NO：17所示；b)HCDR2如SEQ ID NO：18、SEQ ID NO：19、SEQ ID NO：20、SEQ ID NO：21、SEQ ID NO：22、SEQ ID NO：23、SEQ ID NO：24、SEQ ID NO：25、SEQ ID NO：26、SEQ ID NO：27、SEQ ID NO：28、SEQ ID NO：29、SEQ ID NO：30、SEQ ID NO：31、SEQ ID NO：32、SEQ ID NO：33、SEQ ID NO：34、SEQ ID NO：35或SEQ ID NO：36所示；c)HCDR3如SEQ ID NO：37、SEQ ID NO：38、SEQ ID NO：39、SEQ ID NO：40、SEQ ID NO：41、SEQ ID NO：42、SEQ ID NO：43、SEQ ID NO：44、SEQ ID NO：45、SEQ ID NO：46、SEQ ID NO：47、SEQ ID NO：48、SEQ ID NO：49、SEQ ID NO：50、SEQ ID NO：51、SEQ ID NO：52、SEQ ID NO：53、SEQ ID NO：54、SEQ ID NO：55、SEQ ID NO：56、SEQ ID NO：57或SEQ ID NO：58所示；d)LCDR1如SEQ ID NO：59、SEQ ID NO：60、SEQ ID NO：61、SEQ ID NO：62、SEQ ID NO： 63、SEQ ID NO：64、SEQ ID NO：65、SEQ ID NO：66、SEQ ID NO：67、SEQ ID NO：68、SEQ ID NO：69、SEQ ID NO：70、SEQ ID NO：71、SEQ ID NO：72、SEQ ID NO：73、SEQ ID NO：74、SEQ ID NO：75或SEQ ID NO：76所示；e)LCDR2如SEQ ID NO：77、SEQ ID NO：78、SEQ ID NO：79、SEQ ID NO：80、SEQ ID NO：81、SEQ ID NO：82、SEQ ID NO：83、SEQ ID NO：84、SEQ ID NO：85、SEQ ID NO：86或SEQ ID NO：87所示；f)LCDR3如SEQ ID NO：88、SEQ ID NO：89、SEQ ID NO：90、SEQ ID NO：91、SEQ ID NO：92、SEQ ID NO：93、SEQ ID NO：94、SEQ ID NO：95、SEQ ID NO：96、SEQ ID NO：97、SEQ ID NO：98、SEQ ID NO：99、SEQ ID NO：100、SEQ ID NO：101、SEQ ID NO：102、SEQ ID NO：103或SEQ ID NO：104所示。 Ab is an anti-HER3 antibody or an antigen-binding fragment thereof, comprising: an antibody heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 regions, and an antibody light chain variable region comprising LCDR1, LCDR2 and LCDR3 regions, wherein: a) HCDR1 is as shown in SEQ ID NO:01, SEQ ID NO:02, SEQ ID NO:03, SEQ ID NO:04, SEQ ID NO:05, SEQ ID NO:06, SEQ ID NO:07, SEQ ID NO:08, SEQ ID NO:09, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16 or SEQ ID NO:17; b) HCDR2 is as shown in SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25 NO: 24, SEQ ID NO: 25, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35 or SEQ ID NO: 36; c) HCDR3 is as shown in SEQ ID NO: 37, SEQ ID NO: 38 , SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57 or SEQ ID NO: 58; d) LCDR1 as SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 7 2. SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75 or SEQ ID NO: 76; e) LCDR2 is as SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86 or SEQ ID NO: 87; f) LCDR3 as SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO：90、SEQ ID NO：91、SEQ ID NO：92、SEQ ID NO：93、SEQ ID NO：94、SEQ ID NO：95、SEQ ID NO：96、SEQ ID NO：97、SEQ ID NO：98、SEQ ID NO：99、SEQ ID NO：100、SEQ ID NO：101、SEQ ID NO：102、SEQ ID NO：103或SEQ ID NO：104所示。

在一些實施方案中，通式(A)中的Ab的重鏈可變區包含：分別如SEQ ID NO：01所示的HCDR1、如SEQ ID NO：18所示的HCDR2和如SEQ ID NO：37所示的HCDR3；或分別如SEQ ID NO：02所示的HCDR1、如SEQ ID NO：19所示的HCDR2和如SEQ ID NO：38所示的HCDR3；或分別如SEQ ID NO：03所示的HCDR1、如SEQ ID NO：20所示的HCDR2和如SEQ ID NO：39所示的HCDR3；或分別如SEQ ID NO：04所示的HCDR1、如SEQ ID NO：21所示的HCDR2和如SEQ ID NO：40所示的HCDR3；或分別如SEQ ID NO：05所示的HCDR1、如SEQ ID NO：22所示的HCDR2和如SEQ ID NO：41所示的HCDR3；或分別如SEQ ID NO：06所示的HCDR1、如SEQ ID NO：23所示的HCDR2和如SEQ ID NO：42所示的HCDR3；或分別如SEQ ID NO：07 所示的HCDR1、如SEQ ID NO：24所示的HCDR2和如SEQ ID NO：43所示的HCDR3；或分別如SEQ ID NO：06所示的HCDR1、如SEQ ID NO：25所示的HCDR2和如SEQ ID NO：44所示的HCDR3；或分別如SEQ ID NO：06所示的HCDR1、如SEQ ID NO：26所示的HCDR2和如SEQ ID NO：45所示的HCDR3；或分別如SEQ ID NO：08所示的HCDR1、如SEQ ID NO：27所示的HCDR2和如SEQ ID NO：46所示的HCDR3；或分別如SEQ ID NO：09所示的HCDR1、如SEQ ID NO：28所示的HCDR2和如SEQ ID NO：47所示的HCDR3；或分別如SEQ ID NO：10所示的HCDR1、如SEQ ID NO：29所示的HCDR2和如SEQ ID NO：48所示的HCDR3；或分別如SEQ ID NO：11所示的HCDR1、如SEQ ID NO：30所示的HCDR2和如SEQ ID NO：49所示的HCDR3；或分別如SEQ ID NO：12所示的HCDR1、如SEQ ID NO：31所示的HCDR2和如SEQ ID NO：50所示的HCDR3；或分別如SEQ ID NO：01所示的HCDR1、如SEQ ID NO：21所示的HCDR2和如SEQ ID NO：51所示的HCDR3；或分別如SEQ ID NO：06所示的HCDR1、如SEQ ID NO：32所示的HCDR2和如SEQ ID NO：52所示的HCDR3；或分別如SEQ ID NO：13所示的HCDR1、如SEQ ID NO：33所示的HCDR2和如SEQ ID NO：53所示的HCDR3；或分別如SEQ ID NO：14所示的HCDR1、如SEQ ID NO：33所示的HCDR2和如SEQ ID NO：54所示的HCDR3；或分別如SEQ ID NO：15所示的HCDR1、如SEQ ID NO：34所示的HCDR2和如SEQ ID NO：55所示的HCDR3；或分別如SEQ ID NO：16所示的HCDR1、如SEQ ID NO：35所示的HCDR2和如SEQ ID NO：56所示的HCDR3；或分別如SEQ ID NO：17所示的HCDR1、如SEQ ID NO：36所示的HCDR2和如SEQ ID NO：57所示的HCDR3；或分別如SEQ ID NO：06所示的HCDR1、如SEQ ID NO：32所示的HCDR2和如SEQ ID NO：58所示的HCDR3。 In some embodiments, the heavy chain variable region of Ab in general formula (A) comprises: HCDR1 as shown in SEQ ID NO: 01, HCDR2 as shown in SEQ ID NO: 18, and HCDR3 as shown in SEQ ID NO: 37, respectively; or HCDR1 as shown in SEQ ID NO: 02, HCDR2 as shown in SEQ ID NO: 19, and HCDR3 as shown in SEQ ID NO: 38, respectively; or HCDR1 as shown in SEQ ID NO: 03, HCDR2 as shown in SEQ ID NO: 20, and HCDR3 as shown in SEQ ID NO: 39, respectively; or HCDR1 as shown in SEQ ID NO: 04, HCDR2 as shown in SEQ ID NO: 21, and HCDR3 as shown in SEQ ID NO: 40, respectively; or HCDR1 as shown in SEQ ID NO: 05, HCDR2 as shown in SEQ ID NO: 22, and HCDR3 as shown in SEQ ID NO: 41, respectively; or HCDR1 as shown in SEQ ID NO: 06, HCDR2 as shown in SEQ ID NO: 27, and HCDR3 as shown in SEQ ID NO: 38, respectively. NO: 23 and HCDR2 and SEQ ID NO: 42; or SEQ ID NO: 07 HCDR1, SEQ ID NO: 24 and SEQ ID NO: 43; or SEQ ID NO: 06 HCDR1, SEQ ID NO: 25 and SEQ ID NO: 44; or SEQ ID NO: 06 HCDR1, SEQ ID NO: 26 and SEQ ID NO: 45; or SEQ ID NO: 08 HCDR1, SEQ ID NO: 27 and SEQ ID NO: 46; or SEQ ID NO: 09 HCDR1, SEQ ID NO: 28 and SEQ ID NO: 47; or SEQ ID NO: 10 HCDR1, SEQ ID NO: 29 and SEQ ID NO: 30 or HCDR1 as shown in SEQ ID NO:11, HCDR2 as shown in SEQ ID NO:30 and HCDR3 as shown in SEQ ID NO:49, respectively; or HCDR1 as shown in SEQ ID NO:12, HCDR2 as shown in SEQ ID NO:31 and HCDR3 as shown in SEQ ID NO:50, respectively; or HCDR1 as shown in SEQ ID NO:01, HCDR2 as shown in SEQ ID NO:21 and HCDR3 as shown in SEQ ID NO:51, respectively; or HCDR1 as shown in SEQ ID NO:06, HCDR2 as shown in SEQ ID NO:32 and HCDR3 as shown in SEQ ID NO:52, respectively; or HCDR1 as shown in SEQ ID NO:13, HCDR2 as shown in SEQ ID NO:33 and HCDR3 as shown in SEQ ID NO:53, respectively; or HCDR1 as shown in SEQ ID NO:14, HCDR2 as shown in SEQ ID NO:33 and HCDR3 as shown in SEQ ID NO:54, respectively. NO: 54; or HCDR1 as shown in SEQ ID NO: 15, HCDR2 as shown in SEQ ID NO: 34 and HCDR3 as shown in SEQ ID NO: 55; or HCDR1 as shown in SEQ ID NO: 16, HCDR2 as shown in SEQ ID NO: 35 and HCDR3 as shown in SEQ ID NO: 56; or HCDR1 as shown in SEQ ID NO: 17, HCDR2 as shown in SEQ ID NO: 36 and HCDR3 as shown in SEQ ID NO: 57; or HCDR1 as shown in SEQ ID NO: 06, HCDR2 as shown in SEQ ID NO: 32 and HCDR3 as shown in SEQ ID NO: 58.

在一些實施方案中，通式(A)中的Ab的輕鏈可變區包含：分別如SEQ ID NO：59所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：58所示的LCDR3；或分別如SEQ ID NO：60所示的LCDR1、如SEQ ID NO：78所示的LCDR2和如SEQ ID NO：89所示的LCDR3；或分別如SEQ ID NO：61所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：90所示的LCDR3；或分別如SEQ ID NO：59所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：90所示的LCDR3；或分別如SEQ ID NO：62所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：91所示的LCDR3；或分別如SEQ ID NO：63所示的LCDR1、如SEQ ID NO：79所示的LCDR2和如SEQ ID NO：92所示的LCDR3；或分別如SEQ ID NO：60所示的LCDR1、如SEQ ID NO：80所示的LCDR2和如SEQ ID NO：93所示的LCDR3；或分別如SEQ ID NO：64所示的LCDR1、如SEQ ID NO：81所示的LCDR2和如SEQ ID NO：94所示的LCDR3；或分別如SEQ ID NO：65所示的LCDR1、如SEQ ID NO：82所示的LCDR2和如SEQ ID NO：95所示的LCDR3；或分別如SEQ ID NO：60所示的LCDR1、如SEQ ID NO：80所示的LCDR2和如SEQ ID NO：89所示的LCDR3；或分別如SEQ ID NO：66所示的LCDR1、如SEQ ID NO：83所示的LCDR2 和如SEQ ID NO：96所示的LCDR3；或分別如SEQ ID NO：67所示的LCDR1、如SEQ ID NO：79所示的LCDR2和如SEQ ID NO：97所示的LCDR3；或分別如SEQ ID NO：68所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：98所示的LCDR3；或分別如SEQ ID NO：69所示的LCDR1、如SEQ ID NO：84所示的LCDR2和如SEQ ID NO：99所示的LCDR3；或分別如SEQ ID NO：70所示的LCDR1、如SEQ ID NO：85所示的LCDR2和如SEQ ID NO：100所示的LCDR3；或分別如SEQ ID NO：71所示的LCDR1、如SEQ ID NO：85所示的LCDR2和如SEQ ID NO：100所示的LCDR3；或分別如SEQ ID NO：61所示的LCDR1、如SEQ ID NO：77所示的LCDR2和如SEQ ID NO：90所示的LCDR3；或分別如SEQ ID NO：72所示的LCDR1、如SEQ ID NO：86所示的LCDR2和如SEQ ID NO：101所示的LCDR3；或分別如SEQ ID NO：73所示的LCDR1、如SEQ ID NO：82所示的LCDR2和如SEQ ID NO：103所示的LCDR3；或分別如SEQ ID NO：64所示的LCDR1、如SEQ ID NO：82所示的LCDR2和如SEQ ID NO：103所示的LCDR3；或分別如SEQ ID NO：74所示的LCDR1、如SEQ ID NO：79所示的LCDR2和如SEQ ID NO：92所示的LCDR3；或分別如SEQ ID NO：75所示的LCDR1、如SEQ ID NO：82所示的LCDR2和如SEQ ID NO：104所示的LCDR3；或分別如SEQ ID NO：65所示的LCDR1、如SEQ ID NO：82所示的LCDR2和如SEQ ID NO：95所示的LCDR3；或分別如SEQ ID NO：76所示的LCDR1、如SEQ ID NO：85所示的LCDR2和如SEQ ID NO：100所示的LCDR3；或分別如SEQ ID NO：66所示的 LCDR1、如SEQ ID NO：83所示的LCDR2和如SEQ ID NO：96所示的LCDR3。 In some embodiments, the light chain variable region of Ab in Formula (A) comprises: LCDR1 as shown in SEQ ID NO: 59, LCDR2 as shown in SEQ ID NO: 77, and LCDR3 as shown in SEQ ID NO: 58, respectively; or LCDR1 as shown in SEQ ID NO: 60, LCDR2 as shown in SEQ ID NO: 78, and LCDR3 as shown in SEQ ID NO: 89, respectively; or LCDR1 as shown in SEQ ID NO: 61, LCDR2 as shown in SEQ ID NO: 77, and LCDR3 as shown in SEQ ID NO: 90, respectively; or LCDR1 as shown in SEQ ID NO: 59, LCDR2 as shown in SEQ ID NO: 77, and LCDR3 as shown in SEQ ID NO: 90, respectively; or LCDR1 as shown in SEQ ID NO: 62, LCDR2 as shown in SEQ ID NO: 77, and LCDR3 as shown in SEQ ID NO: 91, respectively; or LCDR1 as shown in SEQ ID NO: 63, LCDR2 as shown in SEQ ID NO: 64, LCDR2 as shown in SEQ ID NO: 65, LCDR3 as shown in SEQ ID NO: 66, LCDR3 as shown in SEQ ID NO: 67, LCDR3 as shown in SEQ ID NO: 68, LCDR3 as shown in SEQ ID NO: 69, LCDR1 as shown in SEQ ID NO: 70, LCDR1 as shown in SEQ ID NO: 71, LCDR1 as shown in SEQ ID NO: 72, LCDR1 as shown in SEQ ID NO: 73, LCDR1 as shown in SEQ ID NO: 74, LCDR1 as shown in SEQ ID NO: 75, LCDR1 as shown in SEQ ID NO: 76, LCDR1 as shown in SEQ ID NO: 77, LCDR1 as shown in SEQ ID NO: 78, LCDR1 as shown in SEQ NO:79 LCDR2 and SEQ ID NO:92 LCDR3; or SEQ ID NO:60 LCDR1, SEQ ID NO:80 LCDR2 and SEQ ID NO:93 LCDR3; or SEQ ID NO:64 LCDR1, SEQ ID NO:81 LCDR2 and SEQ ID NO:94 LCDR3; or SEQ ID NO:65 LCDR1, SEQ ID NO:82 LCDR2 and SEQ ID NO:95 LCDR3; or SEQ ID NO:60 LCDR1, SEQ ID NO:80 LCDR2 and SEQ ID NO:89 LCDR3; or SEQ ID NO:66 LCDR1, SEQ ID NO:83 LCDR2 and SEQ ID NO:96 LCDR3; or SEQ ID NO:67 LCDR1, SEQ ID NO:79 LCDR2 and SEQ ID NO:97 LCDR3; or SEQ ID NO:68 LCDR1, SEQ ID NO:79 LCDR2 and SEQ ID NO:98 LCDR3; or SEQ ID NO:69 LCDR2, SEQ ID NO:80 LCDR2 and SEQ ID NO:99 LCDR3; or SEQ ID NO:60 LCDR1, SEQ ID NO:80 LCDR2 and SEQ ID NO:99 LCDR3; or SEQ ID NO:61 LCDR1, SEQ ID NO:79 LCDR2 and SEQ ID NO:99 LCDR3; or SEQ ID NO:67 LCDR1, SEQ ID NO:79 LCDR2 and SEQ ID NO:99 LCDR3; or SEQ ID NO:68 LCDR1, SEQ ID NO:79 LCDR2 and SEQ ID NO:99 LCDR3; or SEQ ID NO:69 ... or LCDR1 as shown in SEQ ID NO:61, LCDR2 as shown in SEQ ID NO:77, and LCDR3 as shown in SEQ ID NO:90; or LCDR1 as shown in SEQ ID NO:69, LCDR2 as shown in SEQ ID NO:84, and LCDR3 as shown in SEQ ID NO:99; or LCDR1 as shown in SEQ ID NO:70, LCDR2 as shown in SEQ ID NO:85, and LCDR3 as shown in SEQ ID NO:100; or LCDR1 as shown in SEQ ID NO:71, LCDR2 as shown in SEQ ID NO:85, and LCDR3 as shown in SEQ ID NO:100; or LCDR1 as shown in SEQ ID NO:61, LCDR2 as shown in SEQ ID NO:77, and LCDR3 as shown in SEQ ID NO:90; or LCDR1 as shown in SEQ ID NO:72, LCDR2 as shown in SEQ ID NO:86, and LCDR3 as shown in SEQ ID NO:87. or LCDR1 as shown in SEQ ID NO:74, LCDR2 as shown in SEQ ID NO:79, and LCDR3 as shown in SEQ ID NO:92; or LCDR1 as shown in SEQ ID NO:75, LCDR2 as shown in SEQ ID NO:82, and LCDR3 as shown in SEQ ID NO:104; or LCDR1 as shown in SEQ ID NO:65, LCDR2 as shown in SEQ ID NO:82, and LCDR3 as shown in SEQ ID NO:95; or LCDR1 as shown in SEQ ID NO:76, LCDR2 as shown in SEQ ID NO:85, and LCDR3 as shown in SEQ ID NO:96. LCDR3 as shown in SEQ ID NO: 100; or LCDR1 as shown in SEQ ID NO: 66, LCDR2 as shown in SEQ ID NO: 83, and LCDR3 as shown in SEQ ID NO: 96.

在較佳的實施方案中，通式(A)中的Ab包含：a)包含分別如SEQ ID NO：08、SEQ ID NO：27和SEQ ID NO：46所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：66、SEQ ID NO：83和SEQ ID NO：96所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或b)包含分別如SEQ ID NO：09、SEQ ID NO：28和SEQ ID NO：47所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：67、SEQ ID NO：79和SEQ ID NO：97所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或c)包含分別如SEQ ID NO：10、SEQ ID NO：29和SEQ ID NO：48所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：68、SEQ ID NO：77和SEQ ID NO：98所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或d)包含分別如SEQ ID NO：11、SEQ ID NO：30和SEQ ID NO：49所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：69、SEQ ID NO：84和SEQ ID NO：99所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或e)包含分別如SEQ ID NO：12、SEQ ID NO：31和SEQ ID NO：50所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：70、SEQ ID NO：85和SEQ ID NO：100所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或f)包含分別如SEQ ID NO：12、SEQ ID NO：31和SEQ ID NO：50所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：71、SEQ ID NO：85和SEQ ID NO：100所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或g)包含分別如SEQ ID NO：01、SEQ ID NO：21和SEQ ID NO：51所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：61、SEQ ID NO：77和SEQ ID NO：90所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或h)包含分別如SEQ ID NO：06、SEQ ID NO：32和SEQ ID NO：52所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：72、SEQ ID NO：86和SEQ ID NO：101所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或i)包含分別如SEQ ID NO：06、SEQ ID NO：32和SEQ ID NO：52所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：73、SEQ ID NO：87和SEQ ID NO：102所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或j)包含分別如SEQ ID NO：13、SEQ ID NO：33和SEQ ID NO：53所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：73、SEQ ID NO：82和SEQ ID NO：103所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或k)包含分別如SEQ ID NO：14、SEQ ID NO：33和SEQ ID NO：54所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：64、SEQ ID NO：82和SEQ ID NO：103所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或l)包含分別如SEQ ID NO：15、SEQ ID NO：34和SEQ ID NO：55所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：74、SEQ ID NO：79和SEQ ID NO：92所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或m)包含分別如SEQ ID NO：16、SEQ ID NO：35和SEQ ID NO：56所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：75、SEQ ID NO：82和SEQ ID NO：104所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或n)包含分別如SEQ ID NO：16、SEQ ID NO：35和SEQ ID NO：56所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：65、SEQ ID NO：82和SEQ ID NO：95所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或o)包含分別如SEQ ID NO：17、SEQ ID NO：36和SEQ ID NO：57所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：76、SEQ ID NO：85和SEQ ID NO：100所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或p)包含分別如SEQ ID NO：06、SEQ ID NO：32和SEQ ID NO：58所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：66、SEQ ID NO：83和SEQ ID NO：96所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或q)包含分別如SEQ ID NO：01、SEQ ID NO：18和SEQ ID NO：37所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：59、SEQ ID NO：77和SEQ ID NO：88所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或r)包含分別如SEQ ID NO：02、SEQ ID NO：19和SEQ ID NO：38所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：60、SEQ ID NO：78和SEQ ID NO：89所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或s)包含分別如SEQ ID NO：03、SEQ ID NO：20和SEQ ID NO：39所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：61、SEQ ID NO：77和SEQ ID NO：90所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或t)包含分別如SEQ ID NO：04、SEQ ID NO：21和SEQ ID NO：40所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：59、SEQ ID NO：77和SEQ ID NO：90所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或u)包含分別如SEQ ID NO：05、SEQ ID NO：22和SEQ ID NO：41所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：62、SEQ ID NO：77和SEQ ID NO：91所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或v)包含分別如SEQ ID NO：06、SEQ ID NO：23和SEQ ID NO：42所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：63、SEQ ID NO：79和SEQ ID NO：92所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或w)包含分別如SEQ ID NO：07、SEQ ID NO：24和SEQ ID NO：43所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：60、SEQ ID NO：80和SEQ ID NO：93所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或x)包含分別如SEQ ID NO：06、SEQ ID NO：25和SEQ ID NO：44所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：64、SEQ ID NO：81和SEQ ID NO：94所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或y)包含分別如SEQ ID NO：06、SEQ ID NO：26和SEQ ID NO：45所示的HCDR1、HCDR2 和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：65、SEQ ID NO：82和SEQ ID NO：95所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列；或z)包含分別如SEQ ID NO：08、SEQ ID NO：27和SEQ ID NO：46所示的HCDR1、HCDR2和HCDR3的重鏈可變區序列；以及包含分別如SEQ ID NO：60、SEQ ID NO：80和SEQ ID NO：89所示的LCDR1、LCDR2和LCDR3的輕鏈可變區序列。 In a preferred embodiment, Ab in general formula (A) comprises: a) a heavy chain variable region sequence comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 08, SEQ ID NO: 27 and SEQ ID NO: 46, respectively; and a light chain variable region sequence comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 66, SEQ ID NO: 83 and SEQ ID NO: 96, respectively; or b) a heavy chain variable region sequence comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 09, SEQ ID NO: 28 and SEQ ID NO: 47, respectively; and a light chain variable region sequence comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 67, SEQ ID NO: 79 and SEQ ID NO: 97, respectively; or c) a heavy chain variable region sequence comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 10, SEQ ID NO: 29 and SEQ ID NO: 11, respectively. NO:48; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:68, SEQ ID NO:77 and SEQ ID NO:98, respectively; or d) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:11, SEQ ID NO:30 and SEQ ID NO:49, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:69, SEQ ID NO:84 and SEQ ID NO:99, respectively; or e) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:12, SEQ ID NO:31 and SEQ ID NO:50, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:70, SEQ ID NO:85 and SEQ ID NO:86, respectively. NO: 100; or f) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 12, SEQ ID NO: 31 and SEQ ID NO: 50, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 71, SEQ ID NO: 85 and SEQ ID NO: 100, respectively; or g) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 01, SEQ ID NO: 21 and SEQ ID NO: 51, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 61, SEQ ID NO: 77 and SEQ ID NO: 90, respectively; or h) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 90, respectively. NO: 52; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 72, SEQ ID NO: 86 and SEQ ID NO: 101, respectively; or i) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 52, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 73, SEQ ID NO: 87 and SEQ ID NO: 102, respectively; or j) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 13, SEQ ID NO: 33 and SEQ ID NO: 53, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 73, SEQ ID NO: 82 and SEQ ID NO: 102, respectively. NO: 103; or k) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 14, SEQ ID NO: 33 and SEQ ID NO: 54, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 64, SEQ ID NO: 82 and SEQ ID NO: 103, respectively; or l) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 15, SEQ ID NO: 34 and SEQ ID NO: 55, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 74, SEQ ID NO: 79 and SEQ ID NO: 92, respectively; or m) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 16, SEQ ID NO: 35 and SEQ ID NO: 93, respectively. NO:56; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:75, SEQ ID NO:82 and SEQ ID NO:104, respectively; or n) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:16, SEQ ID NO:35 and SEQ ID NO:56, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:65, SEQ ID NO:82 and SEQ ID NO:95, respectively; or o) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:17, SEQ ID NO:36 and SEQ ID NO:57, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:76, SEQ ID NO:85 and SEQ ID NO:95, respectively. NO: 100; or p) comprising heavy chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 58, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 66, SEQ ID NO: 83 and SEQ ID NO: 96, respectively; or q) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 01, SEQ ID NO: 18 and SEQ ID NO: 37, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 59, SEQ ID NO: 77 and SEQ ID NO: 88, respectively; or r) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 02, SEQ ID NO: 19 and SEQ ID NO: 100; or p) comprising heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 58, respectively; and comprising light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 66, SEQ ID NO: 83 and SEQ ID NO: 96, respectively; NO:38; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:60, SEQ ID NO:78 and SEQ ID NO:89, respectively; or s) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:03, SEQ ID NO:20 and SEQ ID NO:39, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:61, SEQ ID NO:77 and SEQ ID NO:90, respectively; or t) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:04, SEQ ID NO:21 and SEQ ID NO:40, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:59, SEQ ID NO:77 and SEQ ID NO:90, respectively. NO:90; or u) comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:05, SEQ ID NO:22 and SEQ ID NO:41, respectively; and comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:62, SEQ ID NO:77 and SEQ ID NO:91, respectively; or v) comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO:06, SEQ ID NO:23 and SEQ ID NO:42, respectively; and comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO:63, SEQ ID NO:79 and SEQ ID NO:92, respectively; or w) comprising SEQ ID NO:07, SEQ ID NO:24 and SEQ ID NO: NO: 43; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 60, SEQ ID NO: 80 and SEQ ID NO: 93, respectively; or x) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 25 and SEQ ID NO: 44, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 64, SEQ ID NO: 81 and SEQ ID NO: 94, respectively; or y) heavy chain variable region sequences comprising HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 26 and SEQ ID NO: 45, respectively; and light chain variable region sequences comprising LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 65, SEQ ID NO: 82 and SEQ ID NO: 94, respectively. NO: 95; or z) comprising the heavy chain variable region sequences of HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 08, SEQ ID NO: 27 and SEQ ID NO: 46, respectively; and comprising the light chain variable region sequences of LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 60, SEQ ID NO: 80 and SEQ ID NO: 89, respectively.

在較佳的實施方案中，通式(A)中的Ab選自鼠抗體、嵌合抗體、人源化抗體、人抗體或其抗原結合片段。 In a preferred embodiment, Ab in the general formula (A) is selected from mouse antibodies, chimeric antibodies, humanized antibodies, human antibodies or antigen-binding fragments thereof.

在一些實施方案中，通式(A)中的Ab包含：以下序列的重鏈可變區：SEQ ID NO：127、129、131、133、135、137、139、141、144、145、147、149、151、153、155、105、107、109、111、113、115、117、119、121、123和125，或與其具有至少80%、85%、90%、95%或99%的序列同一性。 In some embodiments, Ab in general formula (A) comprises: a heavy chain variable region of the following sequence: SEQ ID NO: 127, 129, 131, 133, 135, 137, 139, 141, 144, 145, 147, 149, 151, 153, 155, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123 and 125, or having at least 80%, 85%, 90%, 95% or 99% sequence identity therewith.

在一些實施方案中，通式(A)中的Ab包含：以下序列的輕鏈可變區：SEQ ID NO：128、130、132、134、136、138、140、142、143、146、148、150、152、154、106、108、110、112、114、116、118、120、122、124和126，或與其具有至少80%、85%、90%、95%或99%的序列同一性。 In some embodiments, Ab in general formula (A) comprises: a light chain variable region of the following sequence: SEQ ID NO: 128, 130, 132, 134, 136, 138, 140, 142, 143, 146, 148, 150, 152, 154, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124 and 126, or having at least 80%, 85%, 90%, 95% or 99% sequence identity therewith.

在一些實施方案中，通式(A)中的Ab包含：a-1)如SEQ ID NO：127所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：128所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或a-2)如SEQ ID NO：127所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：126所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或b)如SEQ ID NO：129所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：130所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或c)如SEQ ID NO：131所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：132所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或d)如SEQ ID NO：133所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：134所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或e-1)如SEQ ID NO：135所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：136所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或e-2)如SEQ ID NO：137所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：138所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或f)如SEQ ID NO：139所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：140所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或g-1)如SEQ ID NO：141所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如 SEQ ID NO：142所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或g-2)如SEQ ID NO：141所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：143所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或g-3)如SEQ ID NO：144所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：142所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或h)如SEQ ID NO：145所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：146所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或i)如SEQ ID NO：147所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：148所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或j)如SEQ ID NO：149所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：150所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或k-1)如SEQ ID NO：151所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：152所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或k-2)如SEQ ID NO：151所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：122所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或l)如SEQ ID NO：153所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：154所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或m)如SEQ ID NO：155所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：126所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或n)如SEQ ID NO：105所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：106所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或o)如SEQ ID NO：107所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：108所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或p)如SEQ ID NO：109所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：110所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或q)如SEQ ID NO：111所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：112所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或r)如SEQ ID NO：113所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：114所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或s)如SEQ ID NO：115所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：116 所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或t)如SEQ ID NO：117所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：118所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或u)如SEQ ID NO：119所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：120所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或v)如SEQ ID NO：121所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：122所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或w-1)如SEQ ID NO：123所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：124所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或w-2)如SEQ ID NO：125所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：126所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區；或x)如SEQ ID NO：123所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的重鏈可變區；和/或如SEQ ID NO：126所示或與其具有至少80%、85%、90%、95%或99%的序列同一性的輕鏈可變區。 In some embodiments, Ab in general formula (A) comprises: a-1) a heavy chain variable region as shown in SEQ ID NO: 127 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 128 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or a-2) a heavy chain variable region as shown in SEQ ID NO: 127 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 126 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or b) a light chain variable region as shown in SEQ ID NO:129, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:130, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or c) a heavy chain variable region as shown in SEQ ID NO:131, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:132, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or d) a heavy chain variable region as shown in SEQ ID NO:133, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:134, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto. NO:134, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or e-1) a heavy chain variable region as set forth in SEQ ID NO:135, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:136, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or e-2) a heavy chain variable region as set forth in SEQ ID NO:137, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:138, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or f) a heavy chain variable region as set forth in SEQ ID NO:139, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto. NO: 139 or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 140; or g-1) a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 141; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 142; or g-2) a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 141; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: NO:143, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or g-3) a heavy chain variable region as set forth in SEQ ID NO:144, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:142, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or h) a heavy chain variable region as set forth in SEQ ID NO:145, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:146, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or i) a heavy chain variable region as set forth in SEQ ID NO:147, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto. NO:147, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:148, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or j) a heavy chain variable region as shown in SEQ ID NO:149, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:150, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or k-1) a heavy chain variable region as shown in SEQ ID NO:151, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:152, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; NO:152 or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or k-2) a heavy chain variable region as shown in SEQ ID NO:151 or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:122 or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or l) a heavy chain variable region as shown in SEQ ID NO:153 or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:154 or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or m) a heavy chain variable region as shown in SEQ ID NO:155 or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; NO:155, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:126, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or n) a heavy chain variable region as set forth in SEQ ID NO:105, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:106, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or o) a heavy chain variable region as set forth in SEQ ID NO:107, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:110 NO:108, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or p) a heavy chain variable region as set forth in SEQ ID NO:109, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:110, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or q) a heavy chain variable region as set forth in SEQ ID NO:111, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:112, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or r) a heavy chain variable region as set forth in SEQ ID NO:113, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto. NO: 113 or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 114; or s) a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 115; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 116; or t) a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 117; and/or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto as shown in SEQ ID NO: 118. NO:118, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or u) a heavy chain variable region as set forth in SEQ ID NO:119, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:120, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or v) a heavy chain variable region as set forth in SEQ ID NO:121, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as set forth in SEQ ID NO:122, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or w-1) a heavy chain variable region as set forth in SEQ ID NO:1 NO:123, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:124, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or w-2) a heavy chain variable region as shown in SEQ ID NO:125, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:126, or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; or x) a heavy chain variable region as shown in SEQ ID NO:123, or a heavy chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO:1 NO: 126 or a light chain variable region having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto.

在較佳的實施方案中，通式(A)中的Ab包含：a-1)如SEQ ID NO：127所示的重鏈可變區和如SEQ ID NO：128所示的輕鏈可變區；a-2)如SEQ ID NO：127所示的重鏈可變區和如SEQ ID NO：126所示的輕鏈可變區；b)如SEQ ID NO：129所示的重鏈可變區和如SEQ ID NO：130所示的輕鏈可變區；c)如SEQ ID NO：131所示的重鏈可變區和如SEQ ID NO：132所示的輕鏈可變區；d)如SEQ ID NO：133所示的重鏈可變區和如SEQ ID NO：134所示的輕鏈可變區；e-1)如SEQ ID NO：135所示的重鏈可變區和如SEQ ID NO：136所示的輕鏈可變區；e-2)如SEQ ID NO：137所示的重鏈可變區和如SEQ ID NO：138所示的輕鏈可變區；f)如SEQ ID NO：139所示的重鏈可變區和如SEQ ID NO：140所示的輕鏈可變區；g-1)如SEQ ID NO：141所示的重鏈可變區和如SEQ ID NO：142所示的輕鏈可變區；g-2)如SEQ ID NO：141所示的重鏈可變區和如SEQ ID NO：143所示的輕鏈可變區；g-3)如SEQ ID NO：144所示的重鏈可變區和如SEQ ID NO：142所示的輕鏈可變區；h)如SEQ ID NO：145所示的重鏈可變區和如SEQ ID NO：146所示的輕鏈可變區；i)如SEQ ID NO：147所示的重鏈可變區和如SEQ ID NO：148所示的輕鏈可變區；j)如SEQ ID NO：149所示的重鏈可變區和如SEQ ID NO：150所示的輕鏈可變區；k-1)如SEQ ID NO：151所示的重鏈可變區和如SEQ ID NO：152所示的輕鏈可變區；k-2)如SEQ ID NO：151所示的重鏈可變區和如SEQ ID NO：122所示的輕鏈可變區；l)如SEQ ID NO：153所示的重鏈可變區和如SEQ ID NO：154所示的輕鏈可變區；m)如SEQ ID NO：155所示的重鏈可變區和如SEQ ID NO：126所示的輕鏈可變區；n)如SEQ ID NO：105所示的重鏈可變區和如SEQ ID NO：106所示的輕鏈可變區；o)如SEQ ID NO：107所示的重鏈可變區和如SEQ ID NO：108所示的輕鏈可變區；p)如SEQ ID NO：109所示的重鏈可變區和如SEQ ID NO：110 所示的輕鏈可變區；q)如SEQ ID NO：111所示的重鏈可變區和如SEQ ID NO：112所示的輕鏈可變區；r)如SEQ ID NO：113所示的重鏈可變區和如SEQ ID NO：114所示的輕鏈可變區；s)如SEQ ID NO：115所示的重鏈可變區和如SEQ ID NO：116所示的輕鏈可變區；t)如SEQ ID NO：117所示的重鏈可變區和如SEQ ID NO：118所示的輕鏈可變區；u)如SEQ ID NO：119所示的重鏈可變區和如SEQ ID NO：120所示的輕鏈可變區；v)如SEQ ID NO：121所示的重鏈可變區和如SEQ ID NO：122所示的輕鏈可變區；w-1)如SEQ ID NO：123所示的重鏈可變區和如SEQ ID NO：124所示的輕鏈可變區；w-2)如SEQ ID NO：125所示的重鏈可變區和如SEQ ID NO：126所示的輕鏈可變區；x)如SEQ ID NO：123所示的重鏈可變區和如SEQ ID NO：126所示的輕鏈可變區。 In a preferred embodiment, Ab in general formula (A) comprises: a-1) a heavy chain variable region as shown in SEQ ID NO: 127 and a light chain variable region as shown in SEQ ID NO: 128; a-2) a heavy chain variable region as shown in SEQ ID NO: 127 and a light chain variable region as shown in SEQ ID NO: 126; b) a heavy chain variable region as shown in SEQ ID NO: 129 and a light chain variable region as shown in SEQ ID NO: 130; c) a heavy chain variable region as shown in SEQ ID NO: 131 and a light chain variable region as shown in SEQ ID NO: 132; d) a heavy chain variable region as shown in SEQ ID NO: 133 and a light chain variable region as shown in SEQ ID NO: 134; e-1) a heavy chain variable region as shown in SEQ ID NO: 135 and a light chain variable region as shown in SEQ ID NO: 136; NO:136; e-2) a heavy chain variable region as shown in SEQ ID NO:137 and a light chain variable region as shown in SEQ ID NO:138; f) a heavy chain variable region as shown in SEQ ID NO:139 and a light chain variable region as shown in SEQ ID NO:140; g-1) a heavy chain variable region as shown in SEQ ID NO:141 and a light chain variable region as shown in SEQ ID NO:142; g-2) a heavy chain variable region as shown in SEQ ID NO:141 and a light chain variable region as shown in SEQ ID NO:143; g-3) a heavy chain variable region as shown in SEQ ID NO:144 and a light chain variable region as shown in SEQ ID NO:142; h) a heavy chain variable region as shown in SEQ ID NO:145 and a light chain variable region as shown in SEQ ID NO:146; i) a heavy chain variable region as shown in SEQ ID NO:146; NO: 147 and a light chain variable region as shown in SEQ ID NO: 148; j) a heavy chain variable region as shown in SEQ ID NO: 149 and a light chain variable region as shown in SEQ ID NO: 150; k-1) a heavy chain variable region as shown in SEQ ID NO: 151 and a light chain variable region as shown in SEQ ID NO: 152; k-2) a heavy chain variable region as shown in SEQ ID NO: 151 and a light chain variable region as shown in SEQ ID NO: 122; l) a heavy chain variable region as shown in SEQ ID NO: 153 and a light chain variable region as shown in SEQ ID NO: 154; m) a heavy chain variable region as shown in SEQ ID NO: 155 and a light chain variable region as shown in SEQ ID NO: 126; n) a heavy chain variable region as shown in SEQ ID NO: 105 and a light chain variable region as shown in SEQ ID NO: 106; NO: 106; o) the heavy chain variable region shown in SEQ ID NO: 107 and the light chain variable region shown in SEQ ID NO: 108; p) the heavy chain variable region shown in SEQ ID NO: 109 and the light chain variable region shown in SEQ ID NO: 110; q) the heavy chain variable region shown in SEQ ID NO: 111 and the light chain variable region shown in SEQ ID NO: 112; r) the heavy chain variable region shown in SEQ ID NO: 113 and the light chain variable region shown in SEQ ID NO: 114; s) the heavy chain variable region shown in SEQ ID NO: 115 and the light chain variable region shown in SEQ ID NO: 116; t) the heavy chain variable region shown in SEQ ID NO: 117 and the light chain variable region shown in SEQ ID NO: 118; u) the heavy chain variable region shown in SEQ ID NO: 119 and the light chain variable region shown in SEQ ID NO: 120; NO: 119 heavy chain variable region and SEQ ID NO: 120 light chain variable region; v) SEQ ID NO: 121 heavy chain variable region and SEQ ID NO: 122 light chain variable region; w-1) SEQ ID NO: 123 heavy chain variable region and SEQ ID NO: 124 light chain variable region; w-2) SEQ ID NO: 125 heavy chain variable region and SEQ ID NO: 126 light chain variable region; x) SEQ ID NO: 123 heavy chain variable region and SEQ ID NO: 126 light chain variable region.

在一些實施方案中，通式(A)中的Ab是全長抗體，其進一步包含人抗體恆定區；較佳地，該人抗體恆定區的重鏈恆定區選自人IgG1、IgG2、IgG3和IgG4的恆定區及其保守變體，並且該人抗體恆定區的輕鏈恆定區選自人抗體的κ鏈恆定區和λ鏈恆定區及其保守變體；更佳地，該全長抗體包含SEQ ID NO：156的人抗體重鏈恆定區和SEQ ID NO：157的人輕鏈恆定區。 In some embodiments, Ab in general formula (A) is a full-length antibody, which further comprises a human antibody constant region; preferably, the heavy chain constant region of the human antibody constant region is selected from the constant regions of human IgG1, IgG2, IgG3 and IgG4 and conservative variants thereof, and the light chain constant region of the human antibody constant region is selected from the kappa chain constant region and the lambda chain constant region of human antibodies and conservative variants thereof; more preferably, the full-length antibody comprises a human antibody heavy chain constant region of SEQ ID NO: 156 and a human light chain constant region of SEQ ID NO: 157.

在較佳的實施方案中，通式(A)中的Ab包含：a-1)如SEQ ID NO：180所示的重鏈和如SEQ ID NO：181所示的輕鏈；a-2)如SEQ ID NO：180所示的重鏈和如SEQ ID NO：179所示的輕鏈；b)如SEQ ID NO：182所示的重鏈和如SEQ ID NO：183所示的輕鏈；c)如SEQ ID NO：184所示的重鏈和如SEQ ID NO：185所示的輕鏈；g)如SEQ ID NO： 186所示的重鏈和如SEQ ID NO：187所示的輕鏈；e-1)如SEQ ID NO：188所示的重鏈和如SEQ ID NO：189所示的輕鏈；e-2)如SEQ ID NO：190所示的重鏈和如SEQ ID NO：191所示的輕鏈；f)如SEQ ID NO：192所示的重鏈和如SEQ ID NO：193所示的輕鏈；g-1)如SEQ ID NO：194所示的重鏈和如SEQ ID NO：195所示的輕鏈；g-2)如SEQ ID NO：194所示的重鏈和如SEQ ID NO：196所示的輕鏈；g-3)如SEQ ID NO：197所示的重鏈和如SEQ ID NO：195所示的輕鏈；h)如SEQ ID NO：198所示的重鏈和如SEQ ID NO：199所示的輕鏈；i)如SEQ ID NO：200所示的重鏈和如SEQ ID NO：201所示的輕鏈；j)如SEQ ID NO：202所示的重鏈和如SEQ ID NO：203所示的輕鏈；k-1)如SEQ ID NO：204所示的重鏈和如SEQ ID NO：205所示的輕鏈；k-2)如SEQ ID NO：204所示的重鏈和如SEQ ID NO：175所示的輕鏈；l)如SEQ ID NO：206所示的重鏈和如SEQ ID NO：207所示的輕鏈；m)如SEQ ID NO：208所示的重鏈和如SEQ ID NO：179所示的輕鏈；n)如SEQ ID NO：158所示的重鏈和如SEQ ID NO：159所示的輕鏈；o)如SEQ ID NO：160所示的重鏈和如SEQ ID NO：161所示的輕鏈；p)如SEQ ID NO：162所示的重鏈和如SEQ ID NO：163所示的輕鏈；q)如SEQ ID NO：164所示的重鏈和如SEQ ID NO：165所示的輕鏈；r)如SEQ ID NO：166所示的重鏈和如SEQ ID NO：167所示的輕鏈；s)如SEQ ID NO：168所示的重鏈和如SEQ ID NO：169所示的輕鏈；t)如SEQ ID NO：170所示的重鏈和如SEQ ID NO：171所示的輕鏈；u)如SEQ ID NO：172所示的重鏈和如SEQ ID NO：173所示的輕鏈；v)如SEQ ID NO：174所示的重鏈和如SEQ ID NO：175所示的輕鏈；w-1)如SEQ ID NO：176所示的重鏈和如SEQ ID NO：177所示的輕鏈；w-2)如SEQ ID NO：178所示的重鏈和如SEQ ID NO：179所示的輕鏈；x)如SEQ ID NO：176所示的重鏈和如SEQ ID NO：179所示的輕鏈。 In a preferred embodiment, Ab in the general formula (A) comprises: a-1) a heavy chain as shown in SEQ ID NO: 180 and a light chain as shown in SEQ ID NO: 181; a-2) a heavy chain as shown in SEQ ID NO: 180 and a light chain as shown in SEQ ID NO: 179; b) a heavy chain as shown in SEQ ID NO: 182 and a light chain as shown in SEQ ID NO: 183; c) a heavy chain as shown in SEQ ID NO: 184 and a light chain as shown in SEQ ID NO: 185; g) a heavy chain as shown in SEQ ID NO: 186 and a light chain as shown in SEQ ID NO: 187; e-1) a heavy chain as shown in SEQ ID NO: 188 and a light chain as shown in SEQ ID NO: 189; e-2) a heavy chain as shown in SEQ ID NO: 190 and a light chain as shown in SEQ ID NO: 191; f) a heavy chain as shown in SEQ ID NO: 184 and a light chain as shown in SEQ ID NO: 185; g) a heavy chain as shown in SEQ ID NO: 186 and a light chain as shown in SEQ ID NO: 187; e-1) a heavy chain as shown in SEQ ID NO: 188 and a light chain as shown in SEQ ID NO: 189; e-2) a heavy chain as shown in SEQ ID NO: 190 and a light chain as shown in SEQ ID NO: 191; NO: 192 and the light chain shown in SEQ ID NO: 193; g-1) the heavy chain shown in SEQ ID NO: 194 and the light chain shown in SEQ ID NO: 195; g-2) the heavy chain shown in SEQ ID NO: 194 and the light chain shown in SEQ ID NO: 196; g-3) the heavy chain shown in SEQ ID NO: 197 and the light chain shown in SEQ ID NO: 195; h) the heavy chain shown in SEQ ID NO: 198 and the light chain shown in SEQ ID NO: 199; i) the heavy chain shown in SEQ ID NO: 200 and the light chain shown in SEQ ID NO: 201; j) the heavy chain shown in SEQ ID NO: 202 and the light chain shown in SEQ ID NO: 203; k-1) the heavy chain shown in SEQ ID NO: 204 and the light chain shown in SEQ ID NO: NO: 205; k-2) the heavy chain shown in SEQ ID NO: 204 and the light chain shown in SEQ ID NO: 175; l) the heavy chain shown in SEQ ID NO: 206 and the light chain shown in SEQ ID NO: 207; m) the heavy chain shown in SEQ ID NO: 208 and the light chain shown in SEQ ID NO: 179; n) the heavy chain shown in SEQ ID NO: 158 and the light chain shown in SEQ ID NO: 159; o) the heavy chain shown in SEQ ID NO: 160 and the light chain shown in SEQ ID NO: 161; p) the heavy chain shown in SEQ ID NO: 162 and the light chain shown in SEQ ID NO: 163; q) the heavy chain shown in SEQ ID NO: 164 and the light chain shown in SEQ ID NO: 165; r) the heavy chain shown in SEQ ID NO: 166 and the light chain shown in SEQ ID NO: 167; NO: 167; s) the heavy chain shown in SEQ ID NO: 168 and the light chain shown in SEQ ID NO: 169; t) the heavy chain shown in SEQ ID NO: 170 and the light chain shown in SEQ ID NO: 171; u) the heavy chain shown in SEQ ID NO: 172 and the light chain shown in SEQ ID NO: 173; v) the heavy chain shown in SEQ ID NO: 174 and the light chain shown in SEQ ID NO: 175; w-1) the heavy chain shown in SEQ ID NO: 176 and the light chain shown in SEQ ID NO: 177; w-2) the heavy chain shown in SEQ ID NO: 178 and the light chain shown in SEQ ID NO: 179; x) the heavy chain shown in SEQ ID NO: 176 and the light chain shown in SEQ ID NO: 179.

在一些實施方案中，通式(A)中的Ab選自：Fab、Fab'、F(ab')2、可變片段(Fv)、單鏈可變片段(scFv)、二聚結構域V(雙體抗體(diabody))、二硫穩定化Fv(dsFv)和含CDR的肽。 In some embodiments, Ab in the general formula (A) is selected from: Fab, Fab', F(ab')2, variable fragment (Fv), single-chain variable fragment (scFv), dimerization domain V (diabody), disulfide-stabilized Fv (dsFv) and CDR-containing peptide.

在一些實施方案中，細胞毒性藥物選自毒素、化學治療劑、抗生素、放射性同位素和溶核酸酶。 In some embodiments, the cytotoxic drug is selected from toxins, chemotherapeutic agents, antibiotics, radioisotopes, and nucleolytic enzymes.

在另外的實施方案中，細胞毒性藥物選自微管蛋白抑制劑或拓撲異構酶抑制劑；較佳澳瑞他汀(auristatin)類似物或喜樹鹼衍生物；更佳SN-38、MMAE、MMAF或依喜替康(Exatecan)。 In another embodiment, the cytotoxic drug is selected from tubulin inhibitors or topoisomerase inhibitors; preferably auristatin analogs or camptothecin derivatives; more preferably SN-38, MMAE, MMAF or Exatecan.

在一些實施方案中，抗體-藥物綴合物如通式(B)所示： In some embodiments, the antibody-drug conjugate is as shown in the general formula (B):

其中， in,

L₁和L₂是連接單元； _L1 and _L2 are connection units;

y是選自1至10的數，較佳選自2至8的數，更佳2至6或4至8的數，進一步佳6至8的數，並且最佳4、6、7、8； y is a number selected from 1 to 10, preferably a number selected from 2 to 8, more preferably a number from 2 to 6 or 4 to 8, further preferably a number from 6 to 8, and most preferably 4, 6, 7, 8;

Ab是上面提及的抗HER3抗體或抗原結合片段。 Ab is the anti-HER3 antibody or antigen-binding fragment mentioned above.

在較佳的實施方案中，抗體-藥物綴合物如通式(C)所示： In a preferred embodiment, the antibody-drug conjugate is as shown in the general formula (C):

其中， in,

L₂是連接單元； _L2 is the connection unit;

R₁選自氫、C_1-6鹵烷基或C_3-8環烷基； _R1 is selected from hydrogen, _C1-6 halogen alkyl or _C3-8 cycloalkyl;

R₂選自氫、C_1-6鹵烷基或C_3-8環烷基； _R2 is selected from hydrogen, _C1-6 halogen alkyl or _C3-8 cycloalkyl;

或，R₁和R₂與其連接的碳原子一起形成C_3-8環烷基。 Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-8 cycloalkyl group.

其中， in,

L₂是連接單元； _L2 is the connection unit;

R₁選自氫、C_1-3鹵烷基或C_3-6環烷基； _R1 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

R₂選自氫、C_1-3鹵烷基或C_3-6環烷基； _R2 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

或，R₁和R₂與其連接的碳原子一起形成C_3-6環烷基。 Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-6 cycloalkyl group.

在一些實施方案中，連接單元L₂如通式(I)所示： In some embodiments, the linking unit _L2 is represented by the general formula (I):

其中， in,

s¹和s²各自獨立地為選自0-8的整數，較佳地，s¹和s²獨立地選自1、2、3、4、5或6； ^s1 and ^s2 are each independently an integer selected from 0-8, preferably, s1 and ^s2 are independently selected from 1, ² , 3, 4, 5 or 6;

或，s¹是1-8的整數，s²是0，較佳地，s¹選自4、5、6、7或8，並且s²是0； or, ^s1 is an integer from 1 to 8, ^s2 is 0, preferably, ^s1 is selected from 4, 5, 6, 7 or 8, and ^s2 is 0;

或，s²選自2至8的整數，s¹是2，較佳地，s²選自2、3、4、5或6，並且s¹是2。 Or, ^s2 is selected from an integer from 2 to 8, ^s1 is 2, preferably, ^s2 is selected from 2, 3, 4, 5 or 6, and ^s1 is 2.

在較佳的實施方案中，抗體-藥物綴合物如以下結構： In a preferred embodiment, the antibody-drug conjugate has the following structure:

其中， in,

在一方面，抗體-藥物綴合物選自以下化合物： In one aspect, the antibody-drug conjugate is selected from the following compounds:

其中， in,

y是選自1至10的數，較佳選自2至8的數，更佳2至6或4至8的數，進一步佳6至8的數，並且最佳4、6、7、8。 y is a number selected from 1 to 10, preferably a number selected from 2 to 8, more preferably a number from 2 to 6 or 4 to 8, further preferably a number from 6 to 8, and most preferably 4, 6, 7, 8.

本揭露進一步提供了一種用於製備抗體-藥物綴合物或其藥學上可接受的鹽或溶劑化物的方法，該方法包括以下步驟： The present disclosure further provides a method for preparing an antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof, the method comprising the following steps:

在還原後將Ab與化合物偶聯以獲得抗體-藥物綴合物；其中：Ab是上面提及的抗HER3抗體或抗原結合片段； After reduction, Ab is coupled to a compound to obtain an antibody-drug conjugate; wherein: Ab is the anti-HER3 antibody or antigen-binding fragment mentioned above;

或，R₁和R₂與其連接的碳原子一起形成C_3-6環烷基； Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-6 cycloalkyl group;

L₂是上面提及的接頭； _L2 is the connector mentioned above;

在一些實施方案中，本揭露提供了一種醫藥組成物，其包含根據本揭露的配體-藥物綴合物或其藥學上可接受的鹽或溶劑化物，以及一種或多種藥學上可接受的賦形劑、稀釋劑或載體。 In some embodiments, the present disclosure provides a pharmaceutical composition comprising a ligand-drug conjugate according to the present disclosure or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable excipients, diluents or carriers.

在一些實施方案中，本揭露還提供了一種治療或預防與HER3相關的疾病的方法，其包括將治療有效量的根據本揭露的抗體-藥物綴合物或藥學上可接受的鹽或溶劑化物、或醫藥組成物施用於需要治療或預防該疾病的受試者的步驟。 In some embodiments, the present disclosure also provides a method for treating or preventing a disease associated with HER3, comprising the step of administering a therapeutically effective amount of an antibody-drug conjugate or a pharmaceutically acceptable salt or solvate, or a pharmaceutical composition according to the present disclosure to a subject who needs to treat or prevent the disease.

在一些實施方案中，該疾病是具有HER3表達的癌症，較佳地，該癌症是乳腺癌、結直腸癌、肺癌、多發性骨髓瘤、卵巢癌、肝癌、胃癌、胰腺癌、前列腺癌、急性髓系白血病、慢性髓系白血病、骨肉瘤、鱗狀細胞癌、周圍神經鞘瘤、神經鞘瘤、頭頸癌、膀胱癌、食道癌、膠質母細胞瘤、軟組織透明細胞肉瘤、惡性間皮瘤、神經纖維瘤病、腎癌和黑色素瘤。 In some embodiments, the disease is a cancer with HER3 expression, preferably, the cancer is breast cancer, colorectal cancer, lung cancer, multiple myeloma, ovarian cancer, liver cancer, gastric cancer, pancreatic cancer, prostate cancer, acute myeloid leukemia, chronic myeloid leukemia, osteosarcoma, squamous cell carcinoma, peripheral neurilemmoma, neurothecoma, head and neck cancer, bladder cancer, esophageal cancer, glioblastoma, soft tissue clear cell sarcoma, malignant mesothelioma, neurofibromatosis, kidney cancer and melanoma.

在一些實施方案中，本揭露還提供了該抗體-藥物綴合物或藥學上可接受的鹽或溶劑化物和醫藥組成物在製造用於治療或預防與HER3相關的疾病的藥物中的用途。 In some embodiments, the present disclosure also provides the use of the antibody-drug conjugate or pharmaceutically acceptable salt or solvent and pharmaceutical composition in the manufacture of drugs for treating or preventing HER3-related diseases.

在一些實施方案中，本揭露還提供了該抗體-藥物綴合物或藥學上可接受的鹽或溶劑化物和醫藥組成物在製造用於治療或預防具有HER3表達的癌症中的用途；較佳地，該癌症是乳腺癌、結直腸癌、肺癌、多發性骨髓瘤、卵巢癌、肝癌、胃癌、胰腺癌、前列腺癌、急性髓系白血病、慢性髓系白血病、骨肉瘤、鱗狀細胞癌、周圍神經鞘瘤、神經鞘瘤、頭頸癌、膀胱癌、食道癌、膠質母細胞瘤、軟組織透明細胞肉瘤、惡性間皮瘤、神經纖維瘤病、腎癌和黑色素瘤。 In some embodiments, the present disclosure also provides the use of the antibody-drug conjugate or pharmaceutically acceptable salt or solvent and pharmaceutical composition in the manufacture of a method for treating or preventing cancers expressing HER3; preferably, the cancer is breast cancer, colorectal cancer, lung cancer, multiple myeloma, ovarian cancer, liver cancer, gastric cancer, pancreatic cancer, prostate cancer, acute myeloid leukemia, chronic myeloid leukemia, osteosarcoma, squamous cell carcinoma, peripheral neurilemmoma, neurothecoma, head and neck cancer, bladder cancer, esophageal cancer, glioblastoma, soft tissue clear cell sarcoma, malignant mesothelioma, neurofibromatosis, kidney cancer and melanoma.

獲得的抗體或ADC具有一系列優異特徵： The obtained antibodies or ADCs have a series of excellent characteristics:

i)可變區序列與現有抗體不同；我們所有的抗體均是全人抗體，其在人體內引起免疫原性的傾向較小。 i) The variable region sequences are different from existing antibodies; all our antibodies are fully human antibodies, which have a lower tendency to cause immunogenicity in humans.

ii)獲得的抗體具有以高親和力與人結合的能力，這已藉由流式細胞術和ELISA確認。 ii) The obtained antibodies have the ability to bind to humans with high affinity, which has been confirmed by flow cytometry and ELISA.

iii)獲得的抗體具有更好的內化特性用於構建ADC。 iii) The obtained antibodies have better internalization properties and are used to construct ADC.

iv)與帕曲妥單抗-DXd相比，該ADC的有效載荷具有更好的IC₅₀。 iv) The payload of this ADC has a better _IC50 compared to Pertrastuzumab-DXd.

v)當HER3在存在NRG1時與HER2異二聚化時，該ADC的細胞毒性可以不受阻礙。 v) When HER3 heterodimerizes with HER2 in the presence of NRG1, the cytotoxicity of this ADC may not be hindered.

圖1為使用流式細胞術分析對HER3融合瘤株與HER3+(T47D)和HER3-(Jurkat E6.1)細胞系進行體外結合表徵。 FIG1 shows the in vitro binding characterization of HER3 fusion tumor lines to HER3+ (T47D) and HER3- (Jurkat E6.1) cell lines using flow cytometry analysis.

圖2為使用間接殺傷測定對所選擇的融合瘤株的細胞內化活性進行表徵。 FIG2 shows the characterization of the cellular internalization activity of selected fusion tumor lines using an indirect killing assay.

圖3為HER3+ CHO-K1-huHER3細胞中抗HER3重組抗體的內化測定。 Figure 3 shows the internalization assay of anti-HER3 recombinant antibodies in HER3+ CHO-K1-huHER3 cells.

圖4為如藉由CellTiter-Glo發光活力測定所確定的，ADC針對不同腫瘤細胞系(A和B，HCC1569細胞；C，MX-1細胞)的體外細胞毒性活性。 FIG. 4 shows the in vitro cytotoxic activity of ADCs against different tumor cell lines (A and B, HCC1569 cells; C, MX-1 cells) as determined by CellTiter-Glo luminescent viability assay.

圖5為NRG競爭中HER3重組抗體的細胞結合和HER3 ADC殺傷測定。 Figure 5 shows the cell binding and HER3 ADC killing assays of NRG Competition's HER3 recombinant antibodies.

圖6為ADC(3mg/kg，QW)在HCC1569/MX-1腫瘤異種移植小鼠模型中的體內功效。 FIG6 shows the in vivo efficacy of ADC (3 mg/kg, QW) in the HCC1569/MX-1 tumor xenograft mouse model.

圖7為ADC(2mg/kg或6mg/kg，QW)在MX-1腫瘤異種移植小鼠模型中的體內功效。 FIG. 7 shows the in vivo efficacy of ADC (2 mg/kg or 6 mg/kg, QW) in the MX-1 tumor xenograft mouse model.

圖8為ADC(3mg/kg，QW)在MX-1腫瘤異種移植小鼠模型中的體內功效。 FIG8 shows the in vivo efficacy of ADC (3 mg/kg, QW) in the MX-1 tumor xenograft mouse model.

本發明基於開發一種可以與HER3特異性結合的抗體。本發明的抗體可以視需要地與生長抑制劑或細胞毒性劑(如毒素，包括例如依喜替康或依喜替康衍生物)綴合。 The present invention is based on the development of an antibody that can specifically bind to HER3. The antibody of the present invention can be conjugated with a growth inhibitor or a cytotoxic agent (such as a toxin, including, for example, ixotecan or an ixotecan derivative) as desired.

本節中使用的標題僅為方便說明，並不限制本發明。除非本文另有定義，否則本文所用的科學術語和技術術語具有與所屬技術領域中具有通常知識者通常理解的相同含義。此外，除非上下文特別要求，否則單數形式包括複數形式，且複數形式包括單數形式。胺基酸殘基的縮寫是本領域使用的標準三字母和/或一字母代碼，其表示20種常見L-胺基酸之一。 The headings used in this section are for convenience only and do not limit the present invention. Unless otherwise defined herein, the scientific and technical terms used herein have the same meanings as those commonly understood by those of ordinary skill in the art. In addition, unless the context specifically requires otherwise, the singular includes the plural and the plural includes the singular. The abbreviations for amino acid residues are standard three-letter and/or one-letter codes used in the art that represent one of the 20 common L-amino acids.

定義Definition

術語“抗體”是指包含藉由二硫鍵相互連接的至少兩條重(H)鏈和兩條輕(L)鏈的糖蛋白，或其抗原結合部分。每條重鏈由重鏈可變區(本文縮寫為VH)和重鏈恆定區組成。每條輕鏈由輕鏈可變區(本文縮寫為VL)和輕鏈恆定區組成。VH區和VL區可以進一步細分為稱為互補決定區(CDR)的超變區，穿插著稱為框架區(FR)的更保守的區域。每個VH和VL由三個CDR和四個FR組成，從胺基末端至羧基末端按以下順序排列：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。重鏈和輕鏈的可變區含有與抗原相互作用的結合結構域。抗體的恆定區可以介導免疫球蛋白與宿主組織或因子的結合，包括免疫系統的各種細胞(例如效應子細胞)和經典補體系統的第一成分(C1q)。 The term "antibody" refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds, or an antigen-binding portion thereof. Each heavy chain consists of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. Each light chain consists of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The VH and VL regions can be further subdivided into hypervariable regions called complementation determining regions (CDRs), interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs, arranged from amino terminus to carboxyl terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with antigens. The constant regions of antibodies mediate the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component of the classical complement system (C1q).

如本文所用，術語抗體的“抗原結合片段”是指抗體的一個或多個片段，其保留與抗原(例如HER3)特異性結合的能力。已表明可以藉由全長抗體的片段來實現抗體的抗原結合功能。術語抗體的“抗原結合片段”內涵蓋的結合片段的示例包括：(i)Fab片段，由VL、VH、CL和CHI結構域組成的單價片段；(ii)F(ab')2片段，包含在鉸鏈區由二硫鍵連接的兩個Fab片段的二價片段；(iii)Fd片段，其由VH和CHI結構域組成；(iv)Fv片段，其由抗體單個臂的VL和VH結構域組成；(v)dAb片段(Ward等人，(1989)Nature 341：544-546)，其由VH結構域組成；(vi)分離的互補決定區(CDR)；和(vii)兩個或更多個分離的CDR的組合，該CDR可以視需要地由合成接頭連接。此外，儘管Fv片段的兩個結構域VL和VH由分開的基因編碼，但可以使用重組方法藉由合成接頭連接它們，使得能夠將其製成單個蛋白質鏈，其中VL和VH區配對以形成單價分子(稱為單鏈Fv(scFv)；參見例如Bird等人(1988)Science 242：423-426；和Huston 等人(1988)Proc.Natl.Acad.Sci.USÅ 85：5879-5883)。此類單鏈抗體也旨在涵蓋於抗體的術語“抗原結合部分”中。 As used herein, the term "antigen-binding fragment" of an antibody refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., HER3). It has been shown that the antigen-binding function of an antibody can be achieved by a fragment of a full-length antibody. Examples of binding fragments encompassed by the term "antigen-binding fragment" of an antibody include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CHI domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bond at the hinge region; (iii) a Fd fragment, which consists of the VH and CHI domains; (iv) a Fv fragment, which consists of the VL and VH domains of a single arm of the antibody; (v) a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of the VH domain; (vi) isolated complementary determining regions (CDRs); and (vii) a combination of two or more isolated CDRs, which may be optionally linked by a synthetic linker. Furthermore, although the two domains of the Fv fragment, VL and VH, are encoded by separate genes, they can be linked by synthetic linkers using recombinant methods, enabling them to be made into a single protein chain in which the VL and VH regions pair to form a monovalent molecule (referred to as a single-chain Fv (scFv); see, e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. US Å 85:5879-5883). Such single-chain antibodies are also intended to be encompassed by the term "antigen-binding portion" of an antibody.

如本文所用，術語“人抗體”旨在包括具有源自人種系免疫球蛋白序列的可變區和恆定區的抗體。本發明的人抗體可以包含未由人種系免疫球蛋白序列編碼的胺基酸殘基(例如藉由體外隨機或位點特異性誘變或藉由體內體細胞突變而引入的突變)。然而，如本文所用，術語“人抗體”不旨在包括其中源自另一種哺乳動物物種(如小鼠)種系的CDR序列已被移植至人框架序列上的抗體。 As used herein, the term "human antibody" is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present invention may contain amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic cell mutagenesis in vivo). However, as used herein, the term "human antibody" is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (such as a mouse) have been grafted onto human framework sequences.

如本文所用，術語“重組人抗體”包括藉由重組手段製備、表達、產生或分離的所有人抗體，如(a)從人免疫球蛋白基因的轉基因或轉染色體的動物(例如小鼠)或由其製備的融合瘤分離的抗體(進一步描述於以下第I節中)，(b)從經轉化以表達抗體的宿主細胞分離(例如從轉染瘤分離)的抗體，(c)從重組組合人抗體文庫分離的抗體，和(d)藉由涉及將人免疫球蛋白基因序列剪接至其他DNA序列的任何其他方式製備、表達、產生或分離的抗體。此類重組人抗體具有源自人種系免疫球蛋白序列的可變區和恆定區。然而，在某些實施方案中，可以對此類重組人抗體進行體外誘變(或者，當使用人Ig序列轉基因的動物時，進行體內體細胞誘變)，因此重組抗體的VH和VL區的胺基酸序列是這樣的序列，其雖然源自人種系VH和VL序列並與之相關，但可以不是天然存在於體內的人抗體種系貯庫(repertoire)中。 As used herein, the term "recombinant human antibody" includes all human antibodies prepared, expressed, generated or isolated by recombinant means, such as (a) antibodies isolated from animals (e.g., mice) transgenic or transchromosomal for human immunoglobulin genes or fusion tumors prepared therefrom (further described in Section I below), (b) antibodies isolated from host cells transformed to express antibodies (e.g., isolated from transfectomas), (c) antibodies isolated from recombinant combinatorial human antibody libraries, and (d) antibodies prepared, expressed, generated or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies may be subjected to in vitro mutagenesis (or, when using animals transgenic for human Ig sequences, in vivo somatic cell mutagenesis) so that the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist in the human antibody germline repertoire in vivo.

術語“CDR”是指抗體可變結構域內的六個高變區之一，其主要有助於抗原結合。六個CDR最常用的定義之一由Kabat E.A.等人(1991) Sequences of proteins of immunological interest.NIH Publication 91-3242提供。如本文所用，CDR的Kabat定義僅適用於輕鏈可變結構域的CDR1、CDR2和CDR3(LCDR1、LCDR2、LCDR3或L1、L2、L3)，以及重鏈可變結構域的CDR1、CDR2和CDR3(HCDR1、HCDR2、HCDR3或H1、H2、H3)。 The term "CDR" refers to one of the six hypervariable regions within an antibody variable domain that primarily contribute to antigen binding. One of the most commonly used definitions of the six CDRs is provided by Kabat E.A. et al. (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242. As used herein, the Kabat definition of CDR applies only to CDR1, CDR2, and CDR3 (LCDR1, LCDR2, LCDR3 or L1, L2, L3) of the light chain variable domain, and CDR1, CDR2, and CDR3 (HCDR1, HCDR2, HCDR3 or H1, H2, H3) of the heavy chain variable domain.

用於鑑定HCVR和LCVR胺基酸序列內的CDR的方法和技術是本領域公知的，並且可以用於鑑定本文揭露的指定HCVR和/或LCVR胺基酸序列內的CDR。可以用於鑑定CDR邊界的示例性慣例包括，例如，基於抗體的三維結構和CDR環的拓撲結構的Chothia(Chothia等人(1989)Nature 342：877-883)、基於抗體序列變異性的Kabat(Kabat等人，Sequences of Proteins of Immunological Interest，第4版，US Department of Health and Human Services,National Institutes of Health(1987))、AbM(University of Bath)、Contact(University College London)、國際ImMunoGeneTics數據庫(IMGT)(萬維網上imgt.cines.fr/)和基於使用大量晶體結構的親和傳播聚類的North CDR定義。所屬技術領域中具有通常知識者可以容易地鑑定由每種編號系統定義的CDR。 Methods and techniques for identifying CDRs within HCVR and LCVR amino acid sequences are known in the art and can be used to identify CDRs within a given HCVR and/or LCVR amino acid sequence disclosed herein. Exemplary conventions that can be used to identify CDR boundaries include, for example, Chothia (Chothia et al. (1989) Nature 342: 877-883) based on the three-dimensional structure of antibodies and the topological structure of CDR loops, Kabat (Kabat et al., Sequences of Proteins of Immunological Interest, 4th edition, US Department of Health and Human Services, National Institutes of Health (1987)) based on antibody sequence variability, AbM (University of Bath), Contact (University College London), International ImMunoGeneTics Database (IMGT) (on the World Wide Web at imgt.cines.fr/), and North CDR definitions based on affinity propagation clustering using a large number of crystal structures. CDRs defined by each numbering system can be easily identified by one of ordinary skill in the art.

CDR編號的有用比較如下： A useful comparison of CDR numbers is as follows:

如本文所用，術語“核酸分子”是指DNA分子和RNA分子。核酸分子可以是單鏈或雙鏈，但較佳為雙鏈DNA。當核酸被置於與另一個核酸序列的功能關係中時，它是“有效連接”的。例如，如果啟動子或增強子影響編碼序列的轉錄，則該啟動子或增強子與編碼序列有效連接。 As used herein, the term "nucleic acid molecule" refers to DNA molecules and RNA molecules. Nucleic acid molecules can be single-stranded or double-stranded, but are preferably double-stranded DNA. A nucleic acid is "operably linked" when it is placed in a functional relationship with another nucleic acid sequence. For example, if a promoter or enhancer affects the transcription of a coding sequence, the promoter or enhancer is operably linked to a coding sequence.

核酸的製備方法是本領域的常規製備方法。較佳地，其包括以下步驟：藉由基因選殖技術獲得編碼上述蛋白質的核酸分子，或藉由人工全長序列合成的方法獲得編碼上述蛋白質的核酸分子。 The method for preparing nucleic acid is a conventional method in the art. Preferably, it includes the following steps: obtaining a nucleic acid molecule encoding the above protein by gene selection technology, or obtaining a nucleic acid molecule encoding the above protein by artificial full-length sequence synthesis.

所屬技術領域中具有通常知識者知曉，編碼蛋白質胺基酸序列的鹼基序列可以被適當地取代、缺失、改變、插入或添加，以提供多核苷酸同系物。可以藉由在維持抗體活性的範圍內取代、缺失或添加編碼蛋白質序列的基因的一個或多個鹼基來製備本發明的多核苷酸的同系物。 It is known to those skilled in the art that the base sequence encoding the amino acid sequence of a protein can be appropriately substituted, deleted, altered, inserted or added to provide a polynucleotide homologue. The homologue of the polynucleotide of the present invention can be prepared by substituting, deleting or adding one or more bases of the gene encoding the protein sequence within the range of maintaining the antibody activity.

術語“配體”是能夠識別並結合靶細胞相關抗原或受體的大分子化合物。配體的作用是將藥物遞送至與配體結合的靶細胞群體。配體包括但不限於蛋白質激素、凝集素、生長因子、抗體和其他能夠與細胞結合的分子。在本發明的實施方案中，配體表示為Ab。配體中的雜原子和連接單元之間可以形成連接鍵。 The term "ligand" is a macromolecular compound that can recognize and bind to target cell-related antigens or receptors. The role of the ligand is to deliver the drug to the target cell population bound to the ligand. Ligands include but are not limited to protein hormones, lectins, growth factors, antibodies and other molecules that can bind to cells. In the embodiment of the present invention, the ligand is represented by Ab. A connecting bond can be formed between the heteroatom and the linking unit in the ligand.

如本文所用，術語“連接單元”是指在抗體-藥物綴合物(即ADC)中將抗體與藥物連接的部分，其可以是可切割的或不可切割的。可切割接頭(即可斷裂接頭或可生物降解接頭)可以在靶細胞中或靶細胞上被破壞，並由此釋放藥物。在一些實施方案中，本發明的連接單元或接頭具有非常好的穩定性，並且大大降低在遞送至靶標的過程中(例如在血液中)的藥物釋放，從而減少副作用和毒性。在一些具體的實施方案中，本發明的連接單元或接頭選自可切割接頭，如基於二硫鍵的接頭(其在較高的硫醇濃度下在腫瘤細胞中被選擇性破壞)、肽接頭(其在腫瘤細胞中被酶切割)和腙接頭。 As used herein, the term "linker unit" refers to the part that connects the antibody to the drug in the antibody-drug conjugate (i.e., ADC), which can be cleavable or non-cleavable. A cleavable linker (i.e., a cleavable linker or a biodegradable linker) can be destroyed in or on the target cell, thereby releasing the drug. In some embodiments, the linker unit or linker of the present invention has very good stability and greatly reduces the release of the drug during delivery to the target (e.g., in the blood), thereby reducing side effects and toxicity. In some specific embodiments, the linking unit or linker of the present invention is selected from cleavable linkers, such as disulfide bond-based linkers (which are selectively destroyed in tumor cells at higher thiol concentrations), peptide linkers (which are enzymatically cleaved in tumor cells) and hydrazone linkers.

術語“細胞毒性藥物”是指能夠強烈破壞腫瘤細胞正常生長的化學分子。原則上，細胞毒性藥物可以在足夠高的濃度下殺傷腫瘤細胞，但由於缺乏特異性，在殺傷腫瘤細胞時，它也導致正常細胞的細胞凋亡，導致嚴重的副作用。在本發明的實施方案中，細胞毒性藥物表示為D，並且非限制性示例包括微管蛋白抑制劑、DNA烷化劑、酪胺酸激酶抑制劑、拓撲異構酶抑制劑和DNA合成抑制劑，較佳拓撲異構酶抑制劑。 The term "cytotoxic drug" refers to a chemical molecule that can strongly destroy the normal growth of tumor cells. In principle, cytotoxic drugs can kill tumor cells at sufficiently high concentrations, but due to the lack of specificity, when killing tumor cells, it also causes apoptosis of normal cells, resulting in serious side effects. In the embodiment of the present invention, the cytotoxic drug is represented by D, and non-limiting examples include tubulin inhibitors, DNA alkylating agents, tyrosine kinase inhibitors, topoisomerase inhibitors and DNA synthesis inhibitors, preferably topoisomerase inhibitors.

術語“配體-細胞毒性藥物綴合物”是指配體藉由連接單元與生物活性藥物連接。在一些具體的實施方案中，“配體-細胞毒性藥物綴合物”較佳為抗體-藥物綴合物(ADC)，其是指單株抗體或抗體片段藉由連接單元與具有生物活性的細胞毒性藥物連接。 The term "ligand-cytotoxic drug conjugate" refers to a ligand connected to a biologically active drug via a linker unit. In some specific embodiments, the "ligand-cytotoxic drug conjugate" is preferably an antibody-drug conjugate (ADC), which refers to a monoclonal antibody or antibody fragment connected to a biologically active cytotoxic drug via a linker unit.

術語“藥物抗體比(DAR)”是指負載在每個配體上的細胞毒性藥物的平均數量，並且也可以表示為藥物量和抗體量的比值。它可以是整數或非整數。每個配體(Ab)的載藥量範圍可以是1-20個細胞毒性藥物(D)。在本發明的實施方案中，藥物抗體比表示為y，y是選自1至10的數，較佳選自2至8的數，更佳2至4、2至6、4至6、4至8、6至8的數，進一步佳4至8或6至8的數，並且最佳4、5、6、7或8。可以藉由常規方法鑑定偶聯反應後每個ADC分子中藥物的平均數量，如紫外/可見光譜、質譜、ELISA測試和HPLC特徵鑑定。 The term "drug-to-antibody ratio (DAR)" refers to the average amount of cytotoxic drug loaded on each ligand, and can also be expressed as the ratio of the amount of drug to the amount of antibody. It can be an integer or non-integer. The drug loading range of each ligand (Ab) can be 1-20 cytotoxic drugs (D). In an embodiment of the present invention, the drug-to-antibody ratio is expressed as y, y is a number selected from 1 to 10, preferably a number selected from 2 to 8, more preferably 2 to 4, 2 to 6, 4 to 6, 4 to 8, 6 to 8, further preferably 4 to 8 or 6 to 8, and most preferably 4, 5, 6, 7 or 8. The average amount of drug in each ADC molecule after the coupling reaction can be identified by conventional methods, such as UV/visible spectroscopy, mass spectrometry, ELISA testing and HPLC characterization.

術語“烷基”是指飽和脂肪烴基團，其是包含1至20個碳原子的直鏈或支鏈基團，較佳具有1至12個碳原子的烷基，更佳具有1至10個碳原子的烷基，並且最佳具有1至6個碳原子(具有1、2、3、4、5或6個碳原子)的烷基。非限制性示例包括甲基、乙基、正丙基、異丙基、正丁基、異丁基、三級丁基、二級丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基、正庚基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基戊基、2,4-二甲基戊基、2,2-二甲基戊基、 3,3-二甲基戊基、2-乙基戊基、3-乙基戊基、正辛基、2,3-二甲基己基、2,4-二甲基己基、2,5-二甲基己基、2,2-二甲基己基、3,3-二甲基己基、4,4-二甲基己基、2-乙基己基、3-乙基己基、4-乙基己基、2-甲基-2-乙基戊基、2-甲基-3-乙基戊基、N-壬基、2-甲基-2-乙基己基、2-甲基-3-乙基己基、2,2-二乙基戊基、正癸基、3,3-二乙基己基、2,2-二乙基己基，及其各種支鏈異構體。更佳地，烷基是具有1至6個碳原子的低級烷基，並且非限制性示例包括甲基、乙基、正丙基、異丙基、正丁基、異丁基、三級丁基、二級丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。烷基可以是被取代的或未被取代的。被取代時，取代基可以在任何可用的連接點上取代。取代基較佳獨立選自以下的一個或多個基團：烷基、烯基、炔基、烷氧基、烷硫基、烷胺基、鹵素、硫醇、羥基、硝基、氰基、環烷基、雜環基、芳基、雜芳基、環烷氧基、雜環烷氧基、環烷硫基、雜環硫基和氧代。 The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group having 1 to 12 carbon atoms, more preferably an alkyl group having 1 to 10 carbon atoms, and most preferably an alkyl group having 1 to 6 carbon atoms (having 1, 2, 3, 4, 5 or 6 carbon atoms). Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tertiary butyl, di-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, N-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof. More preferably, the alkyl group is a lower alkyl group having 1 to 6 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tertiary butyl, secondary butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituents are preferably independently selected from one or more of the following groups: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocyclicthio and oxo.

術語“鹵烷基”是指被一個或多個鹵素取代的烷基，其中烷基如上所定義。 The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

術語“環烷基”是指飽和或部分不飽和的單環或多環烴取代基，其具有3至20個碳原子，較佳3至12個碳原子，更佳3至10個碳原子，並且最佳3至8個碳原子(具有3、4、5、6、7或8個碳原子)。單環環烷基的非限制性示例包括環丙基、環丁基、環戊基、環戊烯基、環己基、環己烯基、環己二烯基、環庚基、環庚三烯基、環辛基等。多環環烷基包括具有螺環、稠環或橋環的環烷基。 The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent having 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 10 carbon atoms, and most preferably 3 to 8 carbon atoms (having 3, 4, 5, 6, 7 or 8 carbon atoms). Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc. Polycyclic cycloalkyls include cycloalkyls having spiro, fused or bridged rings.

如本文所用，術語“轉染瘤”包括表達抗體的重組真核宿主細胞，如CHO細胞、NS/0細胞、HEK293細胞、植物細胞或真菌(包括酵母細胞)。 As used herein, the term "transfectoma" includes recombinant eukaryotic host cells expressing antibodies, such as CHO cells, NS/0 cells, HEK293 cells, plant cells, or fungi (including yeast cells).

可以藉由常規技術(例如，如PCR擴增或基因組文庫篩選的方法)獲得根據本發明的抗體或其片段的DNA分子的序列。此外，編碼輕鏈和重鏈的序列可以融合在一起，以形成單鏈抗體。 The sequence of the DNA molecule of the antibody or its fragment according to the present invention can be obtained by conventional techniques (e.g., methods such as PCR amplification or genomic library screening). In addition, the sequences encoding the light chain and the heavy chain can be fused together to form a single-chain antibody.

一旦獲得相關序列，就可以使用重組方法批量獲得該相關序列。這通常藉由以下進行：藉由常規方法，將序列選殖至載體中，用載體轉化細胞，然後將相關序列與增殖的宿主細胞分離。 Once the relevant sequence is obtained, it can be obtained in bulk using recombinant methods. This is usually done by cloning the sequence into a vector by conventional methods, transforming cells with the vector, and then isolating the relevant sequence from the proliferating host cells.

此外，可以人工合成相關序列，尤其是當片段長度短時。通常，首先合成數個小片段，然後將其連接在一起以獲得長序列的片段。 In addition, related sequences can be synthesized artificially, especially when the fragment length is short. Usually, several small fragments are synthesized first and then connected together to obtain a long sequence fragment.

目前，可以完全藉由化學合成獲得編碼本發明的抗體(或其片段或其衍生物)的DNA序列。然後可以將DNA序列引入本領域已知的各種現存DNA分子(或例如載體)和細胞中。此外，還可以藉由化學合成將突變引入本發明的蛋白質序列中。 At present, the DNA sequence encoding the antibody (or its fragment or its derivative) of the present invention can be obtained completely by chemical synthesis. The DNA sequence can then be introduced into various existing DNA molecules (or vectors, for example) and cells known in the art. In addition, mutations can also be introduced into the protein sequence of the present invention by chemical synthesis.

一般地，在適合表達根據本發明的抗體的條件下培養獲得的宿主細胞。然後，藉由使用常規的免疫球蛋白純化步驟純化本發明的抗體，例如所屬技術領域中具有通常知識者公知的常規分離純化手段，如蛋白A-瓊脂糖、羥基磷灰石色譜、凝膠電泳、透析、離子交換色譜、疏水色譜、分子篩色譜或親和色譜。 Generally, the obtained host cells are cultured under conditions suitable for expressing the antibodies according to the present invention. The antibodies of the present invention are then purified by using conventional immunoglobulin purification steps, such as conventional separation and purification means known to those skilled in the art, such as protein A-aggregate, hydroxyapatite chromatography, gel electrophoresis, dialysis, ion exchange chromatography, hydrophobic chromatography, molecular screening chromatography or affinity chromatography.

可以藉由常規手段鑑定獲得的單株抗體。例如，可以藉由免疫沉澱或體外結合測定(如放射免疫測定(RIA)或酶聯免疫吸附測定(ELISA))來確定單株抗體的結合特異性。可以藉由例如Scatchard分析(Munson等人，Anal.Biochem.,107：220(1980))確定單株抗體的結合親和力。 The obtained monoclonal antibodies can be identified by conventional means. For example, the binding specificity of the monoclonal antibodies can be determined by immunoprecipitation or in vitro binding assays such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). The binding affinity of the monoclonal antibodies can be determined, for example, by Scatchard analysis (Munson et al., Anal. Biochem., 107: 220 (1980)).

根據本發明的抗體可以在細胞中或細胞膜上表達，或者胞外分泌。如有必要，可以根據重組蛋白的物理、化學和其他特性，藉由各種分離方法來分離並純化重組蛋白。這些方法是所屬技術領域中具有通常知識者眾所周知的。這些方法的示例包括但不限於：常規複性處理、藉由蛋白質沉澱劑處理(如鹽沉澱)、離心、藉由滲透進行細胞裂解、超聲處理、超離心、分子篩色譜(凝膠色譜)、吸附色譜、離子交換色譜、高效液相色譜(HPLC)和任何其他液相色譜，及其組合。 The antibodies according to the present invention can be expressed in cells or on cell membranes, or secreted extracellularly. If necessary, the recombinant protein can be separated and purified by various separation methods according to the physical, chemical and other properties of the recombinant protein. These methods are well known to those with ordinary knowledge in the art. Examples of these methods include, but are not limited to: conventional refolding treatment, treatment with a protein precipitant (such as salt precipitation), centrifugation, cell lysis by osmosis, ultrasonic treatment, ultracentrifugation, molecular screening chromatography (gel chromatography), adsorption chromatography, ion exchange chromatography, high performance liquid chromatography (HPLC) and any other liquid chromatography, and combinations thereof.

術語多肽(例如抗原結合片段、蛋白質或抗體)的“變體”是，與另一多肽序列相比，其中一個或多個胺基酸殘基被插入、缺失、添加和/或取代的多肽，並且包括融合多肽。此外，蛋白質變體包括這樣的蛋白質變體，其藉由蛋白質酶切割、磷酸化或其他轉譯後修飾進行修飾，但保持本文揭露的抗體的生物活性，例如與HER3結合和特異性。變體可以與本文揭露的抗體或其抗原結合片段的序列約99%、98%、97%、96%、95%、94%、93%、92%、91%、90%、89%、88%、87%、86%、85%、84%、83%、82%、81%或80%相同。可以參照以下描述計算同一性或同源性百分比(%)。 The term "variant" of a polypeptide (e.g., an antigen binding fragment, protein, or antibody) is a polypeptide in which one or more amino acid residues are inserted, deleted, added, and/or substituted compared to another polypeptide sequence, and includes fusion polypeptides. In addition, protein variants include protein variants that are modified by protease cleavage, phosphorylation, or other post-translational modifications, but retain the biological activity of the antibodies disclosed herein, such as binding and specificity to HER3. The variant may be about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, or 80% identical to the sequence of the antibody or antigen binding fragment thereof disclosed herein. The identity or homology percentage (%) may be calculated with reference to the following description.

在一個實施方案中，同源性或同一性百分比可以計算為100×[(相同位置)/min(TGA,TGB)]，並且在該式中，TGA、TGB是比較的序列 A和B的殘基數與內部間隙位置之和(Russell等人，J.Mol Biol.,244：332-350(1994))。 In one embodiment, the homology or identity percentage can be calculated as 100×[(same position)/min(TGA, TGB)], and in this formula, TGA, TGB are the sum of the residues and internal gap positions of the compared sequences A and B (Russell et al., J. Mol Biol., 244: 332-350 (1994)).

在本發明中，本發明的抗體還包括其保守變體，這意味著與本發明的抗體的胺基酸序列相比，有多至10個，較佳多至8個，更佳多至5個，最佳多至3個胺基酸被具有相似或相似特性的胺基酸取代以形成多肽。這些保守變體多肽較佳藉由根據表A的胺基酸取代產生。 In the present invention, the antibody of the present invention also includes its conservative variants, which means that compared with the amino acid sequence of the antibody of the present invention, up to 10, preferably up to 8, more preferably up to 5, and most preferably up to 3 amino acids are replaced by amino acids with similar or similar properties to form a polypeptide. These conservative variant polypeptides are preferably produced by amino acid substitution according to Table A.

如本文所用，術語“K_D”(M)旨在指特定抗體-抗原相互作用的解離平衡常數。“K_D”是指解離常數，其由K_d與K_a的比值(即K_d/K_a)獲得，並表示為莫耳濃度(M)。鑑於本揭露，可以使用本領域的方法確定抗體的K_D值。例如，抗體的K_D可以藉由使用表面電漿共振來確定，如藉由使用生物傳感器系統(例如系統)或使用生物層干涉技術(如Octet RED96系統)來確定。 As used herein, the term " _KD " (M) is intended to refer to the dissociation equilibrium constant of a particular antibody-antigen interaction. " _KD " refers to the dissociation constant, which is obtained from the ratio of _Kd to _Ka (i.e., _Kd / _Ka ) and is expressed as a molar concentration (M). In view of the present disclosure, the _KD value of an antibody can be determined using methods in the art. For example, the _KD of an antibody can be determined by using surface plasmon resonance, such as by using a biosensor system (e.g., a system) or using bio-layer interferometry (e.g., an Octet RED96 system).

術語“親和力”是抗體或其抗原結合片段與抗原之間相互作用的強度，它由抗原的特性決定，如抗原的大小、形狀和/或電荷，和抗體或抗原結合片段的CDR序列。本領域已知用於確定親和力的方法，可以參考以下內容。 The term "affinity" is the strength of the interaction between an antibody or its antigen-binding fragment and an antigen, which is determined by the characteristics of the antigen, such as the size, shape and/or charge of the antigen, and the CDR sequence of the antibody or antigen-binding fragment. Methods for determining affinity are known in the art and can be referenced below.

當解離常數(K_D)<10⁶M時，抗體或其抗原結合片段被稱為與其靶標(如抗原)“特異性結合”。當K_D<10⁹M時，抗體與其靶標以“高親和力”特異性結合。 An antibody or antigen-binding fragment thereof is said to "specifically bind" to its target (e.g., antigen) when the dissociation constant ( _KD ) is < ¹⁰⁶ M. When _KD < ¹⁰⁹ M, the antibody specifically binds to its target with "high affinity."

如本文所用，術語“醫藥組成物”旨在指含有本文所述的一種或多種化合物或其生理/藥學上可接受的鹽或其前藥與其他化學成分(如生理/藥學上可接受的載體和賦形劑)的混合物。醫藥組成物的目的是促進對生物體的施用，這有利於活性成分的吸收並發揮生物活性。 As used herein, the term "pharmaceutical composition" is intended to refer to a mixture containing one or more compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs thereof and other chemical components (such as physiologically/pharmaceutically acceptable carriers and excipients). The purpose of the pharmaceutical composition is to facilitate administration to an organism, which is beneficial to the absorption of the active ingredient and the exertion of biological activity.

當應用於動物、人、實驗受試者、細胞、組織、器官或生物液體時，“施用”和“處理”是指使該動物、人、受試者、細胞、組織、器官或生物液體與外源性藥物試劑、治療試劑、診斷試劑或組成物接觸。“施用”和“處理”可以例如指治療方法、藥物代謝動力學方法、診斷方法、研究方法和實驗方法。細胞的處理涵蓋使細胞與試劑接觸，以及使液體與試劑接觸，其中該液體與細胞接觸。“施用”和“處理”也指體外處理和離體處理，例如藉由試劑、診斷劑、結合組成物或藉由另一細胞對細胞進行體外處理和離體處理。當應用於人、獸醫或研究受試者時，“治療”是指治療性治療、預防性或預防措施、研究和診斷應用。 "Administering" and "treating" when applied to an animal, a human, an experimental subject, a cell, a tissue, an organ, or a biological fluid means contacting the animal, human, subject, cell, tissue, organ, or biological fluid with an exogenous pharmaceutical agent, therapeutic agent, diagnostic agent, or composition. "Administering" and "treating" may refer, for example, to therapeutic methods, pharmacokinetic methods, diagnostic methods, research methods, and experimental methods. Treatment of cells encompasses contacting cells with an agent, as well as contacting a fluid with a reagent, wherein the fluid is in contact with cells. "Administering" and "treating" also refer to in vitro and ex vivo treatments, such as the treatment of a cell by a reagent, diagnostic agent, binding composition, or by another cell. "Treatment" when applied to human, veterinary, or research subjects refers to therapeutic treatment, prophylactic or preventative measures, research and diagnostic applications.

此外，本揭露包括用於治療與HER3相關的疾病的藥物，其包含本揭露的抗體、其抗原結合片段或抗體-藥物綴合物作為活性成分。 In addition, the present disclosure includes drugs for treating HER3-related diseases, which contain the antibodies, antigen-binding fragments thereof, or antibody-drug conjugates disclosed herein as active ingredients.

對與HER3相關的疾病沒有限制，只要它是與HER3相關聯的疾病即可，例如，可以藉由結合人HER3來減少由本揭露中揭露的分子誘導的治療反應。因此，當在適合於治療應用的製劑和配方中時，本揭露的分子對於那些患有腫瘤、癌症或傳染病的人非常有用。 There is no limitation on the disease associated with HER3, as long as it is a disease associated with HER3, for example, the therapeutic response induced by the molecules disclosed in the present disclosure can be reduced by binding to human HER3. Therefore, when in a formulation suitable for therapeutic application, the molecules disclosed in the present disclosure are very useful for those suffering from tumors, cancers or infectious diseases.

此外，本揭露涉及用於免疫檢測或測量HER3的方法，用於免疫檢測或測量HER3的試劑，用於免疫檢測或測量表達HER3的細胞的方法，以及用於診斷與HER3陽性細胞相關的疾病的診斷試劑，其包含本揭露的特異性識別人HER3的抗體或抗原結合片段作為活性成分。 In addition, the present disclosure relates to methods for immunodetection or measurement of HER3, reagents for immunodetection or measurement of HER3, methods for immunodetection or measurement of cells expressing HER3, and diagnostic reagents for diagnosing diseases associated with HER3-positive cells, which contain the antibody or antigen-binding fragment of the present disclosure that specifically recognizes human HER3 as an active ingredient.

在本揭露中，用於檢測或確定HER3的量的方法可以是任何已知的方法。例如，它包括免疫檢測或測定。 In the present disclosure, the method for detecting or determining the amount of HER3 may be any known method. For example, it includes immunoassay or determination.

免疫檢測或測定是一種藉由使用標記的抗原或抗體來檢測或確定抗體或抗原的量的方法。免疫檢測或測定的示例包括放射性物質標記的免疫抗體方法(RIA)、酶免疫測定(EIA或ELISA)、螢光免疫測定(FIA)、發光免疫測定、蛋白質印跡方法、物理化學方法等。 Immunoassay or assay is a method of detecting or determining the amount of an antibody or antigen by using a labeled antigen or antibody. Examples of immunoassay or assay include radioactive material labeled immunoantibody method (RIA), enzyme immunoassay (EIA or ELISA), fluorescent immunoassay (FIA), luminescent immunoassay, protein blotting method, physicochemical method, etc.

可以藉由使用本發明的抗體或其抗體片段檢測或測量表達HER3的細胞來診斷上述與HER3陽性細胞相關的疾病。 The above-mentioned diseases associated with HER3-positive cells can be diagnosed by using the antibody of the present invention or its antibody fragment to detect or measure cells expressing HER3.

為了檢測表達多肽的細胞，可以使用已知的免疫檢測，並且較佳可以使用免疫沉澱、螢光細胞染色或免疫組織化學染色等。此外，可以使用FMAT8100HTS系統(Applied Bio system)來使用螢光抗體染色方法等。 In order to detect cells expressing polypeptides, known immunoassays can be used, and preferably immunoprecipitation, fluorescent cell staining, or immunohistochemical staining, etc. can be used. In addition, fluorescent antibody staining methods, etc. can be used using the FMAT8100HTS system (Applied Bio system).

實施例Embodiment

本發明藉由以下具體實施例進一步說明。將理解的是，這些實施例僅用於說明目的，並不旨在限制本發明的範圍。以下實施例中沒有詳細條件的實驗方法一般按照常規條件中描述的條件，如Sambrook.J等人《分子選殖實驗指南》(黃培堂等譯，北京：科學出版社，2002)，或按照製造商推薦的條件(如產品說明書)。除非另有說明，否則百分比和份數均以重量計。除非另有說明，否則以下實施例中使用的實驗材料和試劑均為市售。 The present invention is further illustrated by the following specific examples. It will be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental methods without detailed conditions in the following examples are generally in accordance with the conditions described in the conventional conditions, such as Sambrook.J et al. "Molecular Selection Experiment Guide" (Huang Peitang et al., Beijing: Science Press, 2002), or in accordance with the conditions recommended by the manufacturer (such as product instructions). Unless otherwise stated, percentages and parts are measured by weight. Unless otherwise stated, the experimental materials and reagents used in the following examples are commercially available.

實施例中描述的室溫是本領域的常規室溫，並且一般為10-30℃。 The room temperature described in the embodiment is the conventional room temperature in the art and is generally 10-30°C.

實施例1：免疫和抗體篩選Example 1: Immunization and antibody screening

實施例1-1：免疫原的製備Example 1-1: Preparation of immunogen

重組蛋白抗原huHER3-His和恆河猴HER3-His的組合用於免疫人源化小鼠(Alloy GK MIX品系)。簡要地，10μg是用ATX-Gx小鼠進行的實驗中用於皮下或腹膜內注射的蛋白質抗原的典型量。將抗原與專有佐劑混合用於免疫。對於皮下注射，使用2個部位(每個部位50-100μl)；對於腹膜內注射，我們通常使用200μl。 A combination of recombinant protein antigens huHER3-His and macaque HER3-His was used to immunize humanized mice (Alloy GK MIX strain). Briefly, 10 μg is the typical amount of protein antigen used for subcutaneous or intraperitoneal injection in experiments with ATX-Gx mice. Antigens were mixed with a proprietary adjuvant for immunization. For subcutaneous injections, 2 sites were used (50-100 μl per site); for intraperitoneal injections, we typically use 200 μl.

實施例1-2：免疫Example 1-2: Immunization

免疫方案： Immunization regimen:

藉由雙臂(two-armed)免疫方案獲得抗HER3抗體，使用編碼人免疫球蛋白重鏈可變區和κ輕鏈可變區的基因修飾小鼠藉由RIMMS(多位點重複免疫)方案進行。一組小鼠用重組蛋白抗原huHER3-His(AcroBio，目錄號ER3-H5223)免疫並增強，而另一組小鼠用相同的huHER3-His免疫但用恆河猴HER3-His(Sino Bio，目錄號90043-K08H)增強。藉由HER3特異性免疫測定來監測抗體免疫反應。當達到所需的免疫反應時，從每隻小鼠收穫脾細胞並與小鼠骨髓瘤細胞融合以保持其活力並形成融合瘤細胞，並篩選HER3特異性。 Anti-HER3 antibodies were obtained by a two-armed immunization protocol, using genetically modified mice encoding human immunoglobulin heavy chain variable regions and kappa light chain variable regions by the RIMMS (Repeated Immunization at Multiple Sites) protocol. One group of mice was immunized and boosted with the recombinant protein antigen huHER3-His (AcroBio, catalog number ER3-H5223), while the other group of mice was immunized with the same huHER3-His but boosted with macaque HER3-His (Sino Bio, catalog number 90043-K08H). Antibody immune responses were monitored by HER3-specific immunoassays. When the desired immune response was achieved, spleen cells were harvested from each mouse and fused with mouse myeloma cells to maintain their viability and form fusion tumor cells, and screened for HER3 specificity.

實施例1-3：脾細胞融合Example 1-3: Spleen cell fusion

藉由電融合或PEG融合使脾淋巴細胞和骨髓瘤細胞Sp2/0(ATCC® CRL-158)融合以獲得融合瘤細胞。按照製造商的說明，使用Clonacell^TM HY技術(STEMCELL technologies)進行PEG融合。用於電融合的原代細胞：小鼠骨髓瘤細胞系比值為1：1，用於PEG融合為10：1。 Splenic lymphocytes and myeloma cells Sp2/0 (ATCC® CRL-158) were fused by electrofusion or PEG fusion to obtain fused tumor cells. PEG fusion was performed using Clonacell ^TM HY technology (STEMCELL technologies) according to the manufacturer's instructions. The ratio of primary cells: mouse myeloma cell line for electrofusion was 1:1, and for PEG fusion was 10:1.

實施例1-4：藉由ELISA篩選與HER3蛋白特異性結合的融合瘤株Example 1-4: Screening of fusion tumor lines that specifically bind to HER3 protein by ELISA

使用DuoSet ELISA輔助試劑盒(R&D System，DY008)進行ELISA。用1μg/ml人HER3(Acro Bio，目錄號ER3-H5223)、恆河猴HER3(Sino Bio，目錄號90043-K08H)、小鼠HER3(Acro Bio，目錄號ER3-M52H5)、大鼠HER3(Sino Bio，目錄號80111-r08H)或BSA包被ELISA板過夜。藉由用洗滌緩衝液洗滌板三次來洗去過量未結合蛋白質，然後在室溫封閉1小時。將50μl HER3融合瘤上清液一式兩份加入孔中並孵育1小時。洗去過量未結合抗體，並將50μl 1：20000稀釋的二抗山羊抗小鼠IgG Fc-HRP(ab5870)加入每個孔中再1小時。根據製造商的方案，洗滌板，然後加入50μL化學發光劑(顏色A和顏色B)。使用25μL 2N硫酸停止反應。藉由酶標儀(PerkinElmer)測量樣品在450nm處的光密度。所有測試的株均顯示出與人HER3選擇性結合，但不與BSA結合，證明了HER3特異性。 ELISA was performed using DuoSet ELISA Auxiliary Kit (R&D System, DY008). ELISA plates were coated overnight with 1 μg/ml human HER3 (Acro Bio, catalog number ER3-H5223), rhesus macaque HER3 (Sino Bio, catalog number 90043-K08H), mouse HER3 (Acro Bio, catalog number ER3-M52H5), rat HER3 (Sino Bio, catalog number 80111-r08H) or BSA. Excess unbound protein was washed away by washing the plates three times with wash buffer and then blocked at room temperature for 1 hour. 50 μl of HER3 fusion tumor supernatant was added to the wells in duplicate and incubated for 1 hour. Excess unbound antibody was washed away, and 50 μl of 1:20000 diluted secondary antibody goat anti-mouse IgG Fc-HRP (ab5870) was added to each well for another hour. The plate was washed according to the manufacturer's protocol, and then 50 μL of chemiluminescent agent (color A and color B) was added. The reaction was stopped with 25 μL of 2N sulfuric acid. The optical density of the samples at 450 nm was measured by an enzyme marker (PerkinElmer). All strains tested showed selective binding to human HER3, but not to BSA, demonstrating HER3 specificity.

實施例1-5：藉由流式細胞術篩選與HER3+癌細胞特異性結合的融合瘤株Example 1-5: Screening of fusion tumor lines that specifically bind to HER3+ cancer cells by flow cytometry

使用流式細胞術分析在HER3+細胞系T-47D(ATCC，HTB-133)和HER3-細胞系Jurkat E6.1(ATCC，TIB-152)上對融合瘤上清液進行結合測試。簡要地，將細胞染色緩衝液中的50μL細胞(2×10⁶個細胞/mL)與50μL未稀釋的融合瘤上清液混合。混合物在冰上孵育20min，然後用冰冷的染色緩衝液洗滌兩次。隨後用50μL PE標記的二抗(1：400稀釋，BioLegend，Cat#405307)對細胞進行染色20min。用染色緩衝液洗滌並用4%PFA固定後，藉由流式細胞術分析細胞。純化的抗人HER3抗體用作陽性對照。純化的小鼠IgG1抗體用作同種型對照(R&D Systems，Cat#MAB3481)。圖1顯示了藉由流式細胞術測量的所選擇的細胞結合信號的示例。 Fusion tumor supernatants were tested for binding using flow cytometry analysis on the HER3+ cell line T-47D (ATCC, HTB-133) and the HER3- cell line Jurkat E6.1 (ATCC, TIB-152). Briefly, 50 μL of cells (2×10 ⁶ cells/mL) in cell staining buffer were mixed with 50 μL of undiluted fusion tumor supernatant. The mixture was incubated on ice for 20 min and then washed twice with ice-cold staining buffer. The cells were then stained with 50 μL of PE-labeled secondary antibody (1:400 dilution, BioLegend, Cat#405307) for 20 min. After washing with staining buffer and fixing with 4% PFA, cells were analyzed by flow cytometry. Purified anti-human HER3 antibody was used as a positive control. Purified mouse IgG1 antibody was used as an isotype control (R&D Systems, Cat# MAB3481). Figure 1 shows an example of selected cell binding signals measured by flow cytometry.

實施例1-6：藉由間接殺傷測定篩選具有細胞內化活性的融合瘤株Example 1-6: Screening of fusion tumor lines with cell internalization activity by indirect killing assay

藉由在CHO-K1-huHER3細胞(KYinno，cat#KC-1511)中使用間接殺傷測定來測量來自融合瘤上清液的抗HER3抗體的細胞內化活性。將CHO-K1-huHER3細胞以3000個細胞/孔接種在96孔板中並孵育過夜。用融合瘤培養基稀釋來自每個融合瘤株的融合瘤上清液，並與具有可切割接頭的Fab抗小鼠IgG Fc-MMAF綴合物(Moradec，AM-202AF)混合，然後加入每個孔中。小鼠IgG的終濃度為約10nM、2nM和0.4nM。每個孔中Fab抗小鼠IgG Fc-MMAF綴合物的終濃度為20nM。隨著第二Fab-vc-MMAF的存在，內化的抗體/Fac-vc-MMAF綴合物複合物將釋放細胞毒性有效載荷並殺傷細胞。孵育3天後，藉由Cell Titer Glo 2.0 測定(Promega，G9243)檢測每個孔中的活細胞。純化的抗人HER3抗體用作陽性對照。純化的小鼠IgG1抗體用作同種型對照(Biolegend，Cat#400102)。如圖2所示，用所選擇的融合瘤上清液處理的細胞顯示出活力降低，表明抗體內化。 The cellular internalization activity of anti-HER3 antibodies from fusion tumor supernatants was measured by using an indirect killing assay in CHO-K1-huHER3 cells (KYinno, cat#KC-1511). CHO-K1-huHER3 cells were seeded in 96-well plates at 3000 cells/well and incubated overnight. The fusion tumor supernatant from each fusion tumor strain was diluted with fusion tumor culture medium and mixed with Fab anti-mouse IgG Fc-MMAF conjugate with a cleavable linker (Moradec, AM-202AF) and then added to each well. The final concentration of mouse IgG was approximately 10nM, 2nM and 0.4nM. The final concentration of Fab anti-mouse IgG Fc-MMAF conjugate in each well was 20nM. With the presence of the second Fab-vc-MMAF, the internalized antibody/Fac-vc-MMAF conjugate complex will release the cytotoxic payload and kill the cells. After 3 days of incubation, the viable cells in each well were detected by Cell Titer Glo 2.0 assay (Promega, G9243). Purified anti-human HER3 antibody was used as a positive control. Purified mouse IgG1 antibody was used as an isotype control (Biolegend, Cat#400102). As shown in Figure 2, cells treated with selected fusion tumor supernatants showed reduced viability, indicating antibody internalization.

實施例2：陽性融合瘤株的測序Example 2: Sequencing of positive fusion tumor lines

從陽性融合瘤選殖序列的過程如下。收集對數生長期融合瘤細胞，提取RNA，進行逆轉錄，然後進行VDJ區域擴增。對從每個株擴增的cDNA文庫進行下一代測序。獲得與先導候選物的抗體對應的重鏈和輕鏈可變區DNA序列的胺基酸序列。經可製造性評價，在FR區中進行數個突變，每種抗體重鏈可變區和輕鏈可變區的胺基酸序列及CDR序列如下表所示。VH/VL中CDR的胺基酸殘基根據Kabat和Wu編號系統進行編號和註釋。 The process of selecting sequences from positive fusion tumors is as follows. Fusion tumor cells in logarithmic growth are collected, RNA is extracted, reverse transcribed, and then VDJ region amplification is performed. Next generation sequencing is performed on the cDNA library amplified from each strain. The amino acid sequence of the heavy chain and light chain variable region DNA sequence corresponding to the lead candidate antibody is obtained. After manufacturability evaluation, several mutations are made in the FR region. The amino acid sequence and CDR sequence of the heavy chain variable region and light chain variable region of each antibody are shown in the following table. The amino acid residues of the CDR in VH/VL are numbered and annotated according to the Kabat and Wu numbering system.

實施例3：抗HER3重組抗體的構建和表達Example 3: Construction and expression of anti-HER3 recombinant antibodies

實施例3-1：重組抗體的分子選殖Example 3-1: Molecular cloning of recombinant antibodies

直接合成編碼所選擇株的VH和VL區的cDNA序列，作為具有5'末端框內前導序列(MGWSCIILFLVATATGVHS)的DNA片段。使用NEBuilder DNA組裝選殖試劑盒(New England Biolabs)將這些DNA片段選殖至所選擇的載體中。使用EcoRI位點和NheI位點，將VH區選殖至pFUSE-CHIg_hG1載體(InvivoGen #pfuse-hchg1)中，該VH區與載體中的hIgG1重鏈恆定區在框內。使用AgeI位點和BsiWI位點，將VL區選殖至pFUSE2-CLIg_hk載體(InvivoGen，#pfuse2-hclk)中，該VL區與載體中的hIg κ輕鏈恆定區在框內。 cDNA sequences encoding the VH and VL regions of the selected strains were directly synthesized as DNA fragments with an in-frame leader sequence (MGWSCIILFLVATATGVHS) at the 5' end. These DNA fragments were cloned into the selected vector using the NEBuilder DNA Assembly Cloning Kit (New England Biolabs). The VH region was cloned into the pFUSE-CHIg_hG1 vector (InvivoGen #pfuse-hchg1) using the EcoRI site and the NheI site, and the VH region was in frame with the hIgG1 heavy chain constant region in the vector. The VL region was cloned into the pFUSE2-CLIg_hk vector (InvivoGen, #pfuse2-hclk) using the AgeI site and the BsiWI site, and the VL region was in frame with the hIg kappa light chain constant region in the vector.

抗體的IgG形式揭露為以下重鏈和輕鏈全長。 The IgG form of the antibody reveals the following full length heavy and light chains.

實施例3-2：重組抗體的表達和純化Example 3-2: Expression and purification of recombinant antibodies

重鏈表達質粒和輕鏈質粒使用ExpiFectamine 293轉染試劑盒(ThermoFisher，A14524)共轉染至Expi293F細胞(ThermoFisher，#A14527)中，或使用ExpiFectamine CHO轉染試劑盒(ThermoFisher，A29129)共轉染至ExpiCHO-S細胞(ThermoFisher，#A29127)中。基於製造商的說明，質粒DNA濃度達到每ml懸浮細胞1.0μg，LC：HC載體比例為1：1。在37℃，8% CO₂的軌道振盪器上培養轉染的細胞5至7天。收集條件培養基，並在AKTA Pure 25機器(Cytiva)上使用HiTrap MabSelect SuRe色譜柱(Cytiva，#17549112)純化抗體。沖堤的抗體用Tris緩衝液(pH 9.0)中和並進行PBS緩衝液置換。藉由UV吸收測量產物濃度，並藉由SDS-PAGE和HPLC測定質量。 Heavy chain expression plasmid and light chain plasmid were co-transfected into Expi293F cells (ThermoFisher, #A14527) using ExpiFectamine 293 transfection kit (ThermoFisher, A14524) or into ExpiCHO-S cells (ThermoFisher, #A29127) using ExpiFectamine CHO transfection kit (ThermoFisher, A29129). Based on the manufacturer's instructions, the plasmid DNA concentration was 1.0 μg per ml of suspended cells, and the LC:HC vector ratio was 1:1. Transfected cells were cultured on an orbital shaker at 37°C, 8% _CO2 for 5 to 7 days. Conditioned media were collected and antibodies were purified using HiTrap MabSelect SuRe columns (Cytiva, #17549112) on an AKTA Pure 25 machine (Cytiva). The washed antibodies were neutralized with Tris buffer (pH 9.0) and PBS buffer exchanged. Product concentration was measured by UV absorption and quality was determined by SDS-PAGE and HPLC.

實施例4：藉由流式細胞術對抗HER3重組抗體與HER3Example 4: Comparison of anti-HER3 recombinant antibodies and HER3 by flow cytometry ⁺⁺ 細胞系進行結合表徵Cell lines for binding characterization

使用HER3表達陽性的癌細胞系T-47D細胞，藉由FACS分析確定h1gG1 mAb與細胞表面HER3的結合。 Using HER3-positive cancer cell line T-47D cells, FACS analysis was performed to determine the binding of h1gG1 mAb to cell surface HER3.

實驗步驟參考實施例1-5。 The experimental steps refer to Examples 1-5.

實施例5：抗HER3重組抗體在表達HER3的細胞中的細胞內化的表徵Example 5: Characterization of cellular internalization of anti-HER3 recombinant antibodies in cells expressing HER3

在CHO-K1-huHER3細胞(KYinno，cat#KC-1511)中使用間接螢光內化測定來測量抗HER3重組抗體的細胞內化活性。將CHO-K1-huHER3細胞以20000個細胞/孔接種在96孔板中並孵育過夜。用細胞培養基稀釋重組抗體，並與具有pH依賴性螢光報告基因pHast的Fab抗人Fc-pHast綴合物(Advanced Targeting Systems，PH-01)混合，然後加入每個孔中。抗HER3重組抗體的終濃度為1nM、3nM和9nM。每個孔中Fab抗人Fc-pHast綴合物的終濃度為35nM。隨著第二Fab-pHast的存在，內化的抗體/Fac-pHast綴合物複合物在細胞內的酸性環境中將顯示出增加的螢光。孵育17小時後，使用酶標儀測量來自所有孔的螢光。純化的抗人HER3抗體帕曲妥單抗用作陽性對照。純化的人IgG1抗體用作陰性對照。如圖3所示，在CHO-K1-huHER3細胞中觀察到抗HER3抗體的強烈內化信號。 The cellular internalization activity of anti-HER3 recombinant antibodies was measured using an indirect fluorescence internalization assay in CHO-K1-huHER3 cells (KYinno, cat#KC-1511). CHO-K1-huHER3 cells were seeded in 96-well plates at 20,000 cells/well and incubated overnight. The recombinant antibodies were diluted with cell culture medium and mixed with Fab anti-human Fc-pHast conjugate (Advanced Targeting Systems, PH-01) with the pH-dependent fluorescent reporter gene pHast, and then added to each well. The final concentrations of the anti-HER3 recombinant antibodies were 1nM, 3nM, and 9nM. The final concentration of the Fab anti-human Fc-pHast conjugate in each well was 35nM. With the presence of the second Fab-pHast, the internalized antibody/Fac-pHast conjugate complex will show increased fluorescence in the acidic environment inside the cell. After 17 hours of incubation, fluorescence from all wells was measured using an ELISA reader. Purified anti-human HER3 antibody Pertrastuzumab was used as a positive control. Purified human IgG1 antibody was used as a negative control. As shown in Figure 3, a strong internalization signal of the anti-HER3 antibody was observed in CHO-K1-huHER3 cells.

實施例6：抗HER3重組抗體的表位表徵Example 6: Epitope Characterization of Anti-HER3 Recombinant Antibodies

實施例6-1：與人HER3亞結構域蛋白的ELISA結合Example 6-1: ELISA binding to human HER3 subdomain protein

使用DuoSet ELISA輔助試劑盒(R&D System，DY008)進行ELISA。在4℃用1μg/孔的人HER3蛋白(HER3-His)或人HER3亞結構域1&2(HER3 D1-2，胺基酸20-329)、HER3亞結構域2(HER3 D2，胺基酸185-329)、HER3亞結構域3&4(HER3 D3-4，胺基酸330-643)和HER3亞結構域4(HER3 D4，胺基酸496-643)包被96孔ELISA板過夜。藉由用洗板機用洗滌緩衝液洗滌板三次來洗去過量未結合蛋白質，然後在室溫封閉1小時。將100μl的10μg/mL HER3重組抗體一式兩份加入孔中並在室溫孵育1小時。藉由洗板機洗去過量未結合抗體，並將100μl的1：5000 稀釋的二抗山羊抗人IgG Fc-HRP(ab6858)加入每個孔中以在室溫孵育30min。根據製造商的方案，藉由洗板機再次洗滌板，然後加入100μL化學發光劑(TMB)。使用100μL的2N硫酸停止反應。藉由酶標儀(PerkinElmer)測量樣品在450nm處的光密度。所有測試的重組抗體均與全長人HER3蛋白結合。重組抗體20B5-1顯示出與人HER3亞結構域3-4和亞結構域4結合，表明與亞結構域4結合。所有剩餘重組抗體(參見下表8)均顯示出與人HER3亞結構域1-2和亞結構域1結合，表明與亞結構域1結合。 ELISA was performed using DuoSet ELISA Auxiliary Kit (R&D System, DY008). 96-well ELISA plates were coated with 1 μg/well of human HER3 protein (HER3-His) or human HER3 subdomains 1 & 2 (HER3 D1-2, amino acids 20-329), HER3 subdomain 2 (HER3 D2, amino acids 185-329), HER3 subdomains 3 & 4 (HER3 D3-4, amino acids 330-643) and HER3 subdomain 4 (HER3 D4, amino acids 496-643) at 4°C overnight. Excess unbound proteins were washed away by washing the plates three times with washing buffer using a plate washer and then blocked for 1 hour at room temperature. 100 μl of 10 μg/mL HER3 recombinant antibody was added to the wells in duplicate and incubated at room temperature for 1 hour. Excess unbound antibody was washed away by a plate washer, and 100 μl of a 1:5000 dilution of secondary antibody goat anti-human IgG Fc-HRP (ab6858) was added to each well for incubation at room temperature for 30 min. The plate was washed again by a plate washer according to the manufacturer's protocol, and 100 μL of chemiluminescent reagent (TMB) was added. The reaction was stopped using 100 μL of 2N sulfuric acid. The optical density of the samples was measured at 450 nm by an ELISA reader (PerkinElmer). All tested recombinant antibodies bound to the full-length human HER3 protein. Recombinant antibody 20B5-1 showed binding to human HER3 subdomains 3-4 and subdomain 4, indicating binding to subdomain 4. All remaining recombinant antibodies (see Table 8 below) showed binding to human HER3 subdomains 1-2 and subdomain 1, indicating binding to subdomain 1.

實施例6-2：Octet分箱(binning)Example 6-2: Octet binning

對前三個株(18E11-1、20E1-3、23F6-1)進行表位分箱，並與參考抗HER3抗體帕曲妥單抗進行比較。使用配備有來自Pall Life Sciences(Menlo Park,CA)的Ni-NTA生物傳感器的Octet Red384系統進行抗體表位分箱。實驗作為串聯分箱測定進行。測定由五步結合循環組成：1)建立緩衝基線30秒，2)使用由10×動力學緩衝液儲備液(ForteBio)稀釋的標準1×測定緩衝液(PBS+0.02%吐溫20，0.1% BSA，0.05%疊氮化鈉)，將5nM HER3抗原(HER3-His)與Ni-NTA octet傳感器偶聯5分鐘，3)加載25nM每種抗體(飽和mAb)以使固定化抗原飽和10分鐘，4)使25nM每種抗體(競爭mAb)結合5分鐘，和5)使捕獲傳感器再生30秒。如表9和表10所示，所有四種抗HER3抗體可以分為2個不同的表位箱(bin)。118E11-1、20E1-3和23F6-1在同一箱中，而帕曲妥單抗在不同箱中。在18E11-1、20E1-3或23F6-1結合之後，帕曲妥單抗仍然可以與HER3結合，並具有很好的締合曲線，表明表位不同。 The first three strains (18E11-1, 20E1-3, 23F6-1) were epitope binned and compared to the reference anti-HER3 antibody Pertrastuzumab. Antibody epitope binning was performed using the Octet Red 384 system equipped with a Ni-NTA biosensor from Pall Life Sciences (Menlo Park, CA). The experiment was performed as a tandem binning assay. The assay consisted of a five-step binding cycle: 1) establish a buffer baseline for 30 seconds, 2) couple 5 nM HER3 antigen (HER3-His) to the Ni-NTA octet sensor for 5 minutes using standard 1× assay buffer (PBS + 0.02% Tween 20, 0.1% BSA, 0.05% sodium azide) diluted from 10× kinetic buffer stock (ForteBio), 3) load 25 nM of each antibody (saturated mAb) to saturate the immobilized antigen for 10 minutes, 4) allow 25 nM of each antibody (competing mAb) to bind for 5 minutes, and 5) regenerate the capture sensor for 30 seconds. As shown in Tables 9 and 10, all four anti-HER3 antibodies can be divided into 2 different epitope bins. 118E11-1, 20E1-3, and 23F6-1 are in the same bin, while Pertuzumab is in a different bin. After binding of 18E11-1, 20E1-3, or 23F6-1, Pertuzumab can still bind to HER3 with a good binding curve, indicating that the epitopes are different.

實施例7：抗HER3抗體藥物綴合物(ADC)的生成Example 7: Generation of anti-HER3 antibody-drug conjugates (ADCs)

本發明的抗體具有細胞親和力活性和內吞活性，使其適合與藥物偶聯以形成抗體藥物綴合物，用於治療HER3介導的疾病。 The antibody of the present invention has cell affinity activity and endocytosis activity, making it suitable for coupling with drugs to form antibody-drug conjugates for the treatment of HER3-mediated diseases.

單株抗體的純化Purification of monoclonal antibodies

參考實施例3-2中描述的抗HER3抗體的製備程序，純化了單株抗體用於藥物綴合。用於綴合的進一步的抗體可以包括本文描述的任何抗體(參見實施例3)。 Referring to the preparation procedure of the anti-HER3 antibody described in Example 3-2, the monoclonal antibody was purified for drug conjugation. Further antibodies used for conjugation may include any antibody described herein (see Example 3).

作為藥物的中間體的製備Preparation of intermediates for drugs

以下中間體化合物用於生成抗HER3抗體的抗體藥物綴合物(ADC)，化合物D藉由PCT專利申請中揭露的方法製備。(參見2022年1月26日提交的WO2022161385)。Deruxtecan(MedChemExpress，Cat#HY-114233)是陽性對照。藉由PCT專利申請中揭露的方法製備HR9106。(參見2019年9月25日提交的WO2020063673)。 The following intermediate compounds are used to generate antibody-drug conjugates (ADCs) of anti-HER3 antibodies. Compound D is prepared by the method disclosed in the PCT patent application. (See WO2022161385 filed on January 26, 2022). Deruxtecan (MedChemExpress, Cat#HY-114233) is a positive control. HR9106 is prepared by the method disclosed in the PCT patent application. (See WO2020063673 filed on September 25, 2019).

單株抗體與藥物分子的綴合Conjugation of monoclonal antibodies with drug molecules

在37℃用足夠莫耳當量的(三(2-羧基乙基)膦(TCEP，10mM)處理抗HER3抗體(5mg/mL的PBS溶液，pH 7.4)1小時。抗體B12是陰性對照，帕曲妥單抗和MOTA是陽性對照。將足夠莫耳當量的藥物接頭(例如Deruxtecan或化合物D的DMSO溶液)加入還原抗體的PBS溶液中。然後在孵育一小時後，使用體積排阻色譜(SEC)純化反應混合物以分離ADC和游離的未綴合藥物接頭。 Anti-HER3 antibody (5 mg/mL in PBS, pH 7.4) was treated with sufficient molar equivalents of tris(2-carboxyethyl)phosphine (TCEP, 10 mM) for 1 hour at 37°C. Antibody B12 was a negative control, and Pertuzumab and MOTA were positive controls. Sufficient molar equivalents of drug linker (e.g., Deruxtecan or Compound D in DMSO) were added to the reduced antibody in PBS. After incubation for one hour, the reaction mixture was purified using size exclusion chromatography (SEC) to separate ADC and free unconjugated drug linker.

使用TOF LC/MS系統(Agilent)確定ADC的藥物抗體比(DAR)，並且平均DAR值總結於表10中。與化合物D綴合的抗HER3抗體的平均DAR值為大約6.0或7.0。與Deruxtecan綴合的抗HER3抗體的平均DAR值為大約8.0。具有其他DAR值的與化合物D綴合的抗HER3抗體的合成步驟可以參考以上。 The drug-antibody ratio (DAR) of the ADC was determined using a TOF LC/MS system (Agilent), and the average DAR values are summarized in Table 10. The average DAR value of the anti-HER3 antibody conjugated with Compound D was about 6.0 or 7.0. The average DAR value of the anti-HER3 antibody conjugated with Deruxtecan was about 8.0. The synthesis steps of the anti-HER3 antibody conjugated with Compound D having other DAR values can be referred to above.

實施例8：ADC對腫瘤細胞生長的抑制效果Example 8: Inhibitory effect of ADC on tumor cell growth

使用體外細胞殺傷測定來檢查ADC對腫瘤細胞生長的抑制效果。在對數期生長中收集不同腫瘤細胞，並以1000-1500個細胞/孔分佈到96孔板中。孵育過夜後，將ADC加入每個孔中。孔中ADC的終濃度範圍為0.01nM至666.7nM。孵育4至7天後，藉由Cell Titer Glo 2.0測定(Promega)來確定每個孔中的細胞活力。在GraphPad Prism中使用S形劑量-反應非線性回歸擬合來生成曲線和IC₅₀值。來自這些實驗的數據總結於表11A和表11B中。圖4A至圖4C顯示了針對指定細胞系測試的所有ADC的代表性殺傷曲線。所有測試的ADC對腫瘤細胞均顯示出良好的體外毒性，其中ADC-02、ADC-03、ADC-04、ADC-07、ADC-10和ADC-11顯示出優於參考ADC的最顯著毒性。 In vitro cell killing assays were used to examine the inhibitory effect of ADC on tumor cell growth. Different tumor cells were collected during logarithmic growth and distributed into 96-well plates at 1000-1500 cells/well. After overnight incubation, ADC was added to each well. The final concentration range of ADC in the well was 0.01nM to 666.7nM. After 4 to 7 days of incubation, the cell viability in each well was determined by Cell Titer Glo 2.0 assay (Promega). S-shaped dose-response nonlinear regression fitting was used in GraphPad Prism to generate curves and IC ₅₀ values. The data from these experiments are summarized in Tables 11A and 11B. Figures 4A to 4C show representative kill curves for all ADCs tested against the indicated cell lines. All tested ADCs showed good in vitro toxicity against tumor cells, with ADC-02, ADC-03, ADC-04, ADC-07, ADC-10, and ADC-11 showing the most significant toxicity over the reference ADC.

實施例9：在存在NRG1時，與參考ADC不同，HER3 ADC在腫瘤細胞中顯示出細胞毒性Example 9: HER3 ADC shows cytotoxicity in tumor cells in the presence of NRG1, unlike the reference ADC

已顯示NRG1作為天然配體對HER3具有高親和力([Kd]<100pM)(Am J Respir Cell Mol Biol.2000年4月；22(4)：432-40.)，抗體難以與其競爭。然而，抗體可以在不同表位上結合抗原，該不同表位可以與NRG1可以結合之處不同，然後我們在具有不同HER3異二聚體狀態的不同細胞系上測試了我們的抗體。藉由流式細胞術，我們發現18E11-1、20B5-1、20E1-3和23F6-1的結合信號僅受到SKBr3細胞上相等莫耳濃度的NRG1分子的輕微抑制，而帕曲妥單抗的結合受到顯著影響。我們進一步檢查了結合的差異是否會影響HER3 ADC的細胞毒性，並且我們確實觀察到，當NRG1呈現於SKBr3和HCC1569細胞上時，ADC-07和ADC-10表現出優越的細胞毒性。 NRG1 has been shown to have a high affinity ([Kd] < 100 pM) as a natural ligand for HER3 (Am J Respir Cell Mol Biol. 2000 Apr; 22(4): 432-40.), which is difficult for antibodies to compete with. However, antibodies can bind antigens at different epitopes that may be different from those to which NRG1 can bind, and we then tested our antibodies on different cell lines with different HER3 heterodimer states. By flow cytometry, we found that the binding signals of 18E11-1, 20B5-1, 20E1-3, and 23F6-1 were only slightly inhibited by equal molar concentrations of NRG1 molecules on SKBr3 cells, while the binding of pascatalum was significantly affected. We further examined whether differences in binding would affect the cytotoxicity of the HER3 ADCs, and we indeed observed that ADC-07 and ADC-10 exhibited superior cytotoxicity when NRG1 was presented on SKBr3 and HCC1569 cells.

實施例10：ADC對HER3陽性癌細胞裸鼠皮下移植腫瘤模型的腫瘤抑制實驗Example 10: Tumor inhibition experiment of ADC on HER3-positive cancer cell subcutaneous transplanted tumor model in nude mice

1、源自HCC1569細胞系的異種移植(CDX)模型 1. Xenotransplantation (CDX) model derived from HCC1569 cell line

將HCC1569細胞皮下移植至小鼠(CB17/SCID)中。當腫瘤體積達到大約120-180mm³時，將移植小鼠隨機分為四組(每組5隻小鼠)。組別為載體對照、ADC-01、ADC-03和ADC-04。用ADC(3mg/kg)靜脈內QW處理小鼠。利用以下公式計算平均腫瘤生長抑制(TGI)： HCC1569 cells were subcutaneously transplanted into mice (CB17/SCID). When the tumor volume reached approximately 120-180 ^mm3 , the transplanted mice were randomly divided into four groups (5 mice per group). The groups were vector control, ADC-01, ADC-03, and ADC-04. Mice were treated with ADC (3 mg/kg) intravenously QW. The average tumor growth inhibition (TGI) was calculated using the following formula:

TGI=((平均值(C)-平均值(C0))-(平均值(T)-平均值(T0)))/(平均值(C)-平均值(C0))* 100%；T是當前組值，C是對照組值，T0和C0表示測試開始時的腫瘤體積。 TGI=((mean value(C)-mean value(C0))-(mean value(T)-mean value(T0)))/(mean value(C)-mean value(C0))* 100%; T is the current group value, C is the control group value, T0 and C0 represent the tumor volume at the beginning of the test.

用三種ADC處理的小鼠中的腫瘤生長與對照組中的小鼠顯示出統計學上顯著的差異，導致腫瘤消退。 Tumor growth in mice treated with the three ADCs showed statistically significant differences from mice in the control group, resulting in tumor regression.

研究的結果示於表13和圖6A中。用ADC-01、ADC-03和ADC-04處理的小鼠中的腫瘤生長與用載體處理的小鼠顯示出統計學上顯著的差異，第28天的TGI值分別為40.2%、105.2%和105.2%。 The results of the study are shown in Table 13 and Figure 6A. Tumor growth in mice treated with ADC-01, ADC-03, and ADC-04 showed statistically significant differences from that in mice treated with vehicle, with TGI values on day 28 of 40.2%, 105.2%, and 105.2%, respectively.

2、源自MX-1細胞系的異種移植(CDX)模型 2. Xenotransplantation (CDX) model derived from MX-1 cell line

將MX-1細胞皮下移植至小鼠(CB17/SCID)中。當腫瘤體積達到大約120-180mm³時，將移植小鼠隨機分為四組(每組5隻小鼠)。組別為載體對照、ADC-01、ADC-03和ADC-04。用ADC(3mg/kg)靜脈內QW處理小鼠。利用以下公式計算平均腫瘤生長抑制(TGI)： MX-1 cells were subcutaneously transplanted into mice (CB17/SCID). When the tumor volume reached approximately 120-180 ^mm3 , the transplanted mice were randomly divided into four groups (5 mice per group). The groups were vehicle control, ADC-01, ADC-03, and ADC-04. Mice were treated with ADC (3 mg/kg) intravenously QW. The average tumor growth inhibition (TGI) was calculated using the following formula:

研究的結果示於表14和圖6B中。用ADC-04處理的小鼠中的腫瘤生長與用載體處理的小鼠顯示出統計學上顯著的差異，第28天的TGI值分別為5.0%、48.7%和88.4%。 The results of the study are shown in Table 14 and Figure 6B. Tumor growth in mice treated with ADC-04 showed statistically significant differences from those treated with vehicle, with TGI values on day 28 of 5.0%, 48.7%, and 88.4%, respectively.

在相同的MX-1異種移植小鼠模型中進行了ADC-01、ADC-02和ADC-03和ADC-04的追蹤腫瘤抑制研究。將MX-1移植小鼠隨機分為八組(每組5隻小鼠)。組別為載體對照、3mg/kg ADC-01、6mg/kg ADC-02、6mg/kg ADC-03、6mg/kg ADC-04、2mg/kg ADC-02、2mg/kg ADC-03和2mg/kg ADC-04。用ADC靜脈內QW處理小鼠。 Follow-up tumor inhibition studies of ADC-01, ADC-02, ADC-03, and ADC-04 were performed in the same MX-1 xenograft mouse model. MX-1 xenograft mice were randomly divided into eight groups (5 mice per group). Groups were vehicle control, 3mg/kg ADC-01, 6mg/kg ADC-02, 6mg/kg ADC-03, 6mg/kg ADC-04, 2mg/kg ADC-02, 2mg/kg ADC-03, and 2mg/kg ADC-04. Mice were treated with ADC intravenously QW.

研究的結果示於表15和圖7中。用ADC-02、ADC-04以2和6mg/kg處理的小鼠中的腫瘤生長與用ADC-01以3mg/kg處理的小鼠顯示出統計學上顯著的差異，TGI值分別為111.9%、91.0%、113.2%和95.2%，而用ADC-01以3mg/kg處理的小鼠中的腫瘤表現出TGI為58.8%。 The results of the study are shown in Table 15 and Figure 7. Tumor growth in mice treated with ADC-02, ADC-04 at 2 and 6 mg/kg showed statistically significant differences from mice treated with ADC-01 at 3 mg/kg, with TGI values of 111.9%, 91.0%, 113.2% and 95.2%, respectively, while tumors in mice treated with ADC-01 at 3 mg/kg showed a TGI of 58.8%.

ADC-02、ADC-04、ADC-06、ADC-13和ADC-14在相同的MX-1異種移植小鼠模型中進行。將MX-1移植小鼠隨機分為六組(每組5隻小鼠)。組別為載體對照、ADC-02、ADC-04、ADC-06、ADC-13和ADC-14。用ADC(3mg/kg)靜脈內QW處理小鼠3週。 ADC-02, ADC-04, ADC-06, ADC-13, and ADC-14 were performed in the same MX-1 xenograft mouse model. MX-1 xenograft mice were randomly divided into six groups (5 mice per group). The groups were vehicle control, ADC-02, ADC-04, ADC-06, ADC-13, and ADC-14. Mice were treated with ADC (3 mg/kg) intravenously QW for 3 weeks.

研究的結果示於表16和圖8中。具有不同DAR值的ADC-02和ADC-04顯示出相當的腫瘤生長抑制。 The results of the study are shown in Table 16 and Figure 8. ADC-02 and ADC-04 with different DAR values showed comparable tumor growth inhibition.

實施例11：抗HER3抗體和抗體-藥物綴合物在小鼠中的藥物代謝動力學(PK)測定Example 11: Drug metabolism kinetics (PK) determination of anti-HER3 antibodies and antibody-drug conjugates in mice

雌性Balb/C小鼠(6-8週齡)用於評估抗體-藥物綴合物的藥物代謝動力學。在給藥前收集空白血清。以3mg/kg靜脈內施用ADC。注射後，在預定時間點(1、4、24、48、72、120、168、336和504小時)，從指定小鼠收集血清並立即儲存於-80℃直至進一步分析。 Female Balb/C mice (6-8 weeks of age) were used to evaluate the pharmacokinetic of antibody-drug conjugates. Blank serum was collected before dosing. ADC was administered intravenously at 3 mg/kg. After injection, serum was collected from designated mice at predetermined time points (1, 4, 24, 48, 72, 120, 168, 336, and 504 hours) and immediately stored at -80°C until further analysis.

藉由ELISA測量ADC的血清濃度，藉由PK Solver 2.0計算藥物代謝動力學參數。主要藥物代謝動力學參數示於表16和表17中。 The serum concentration of ADC was measured by ELISA, and the pharmacokinetic parameters were calculated by PK Solver 2.0. The main pharmacokinetic parameters are shown in Table 16 and Table 17.

HER3抗體ADC-02和ADC-04在小鼠中的AUC相似。ADC-02和ADC-04的消除半衰期優於ADC-01的消除半衰期，這可以有助於其優越的體內治療功效。 The AUCs of HER3 antibodies ADC-02 and ADC-04 in mice are similar. The elimination half-life of ADC-02 and ADC-04 is superior to that of ADC-01, which may contribute to their superior in vivo therapeutic efficacy.

TW202432185A_112141379_SEQL.xmlTW202432185A_112141379_SEQL.xml

Claims

An antibody-drug conjugate of general formula (A) or a pharmaceutically acceptable salt or solvate thereof,

Ab-(LAb-(L ₂₂ -L-L ₁₁ -D)-D) _yy (A)(A)

in,

D is a cytotoxic drug;

_L1 and _L2 are connection units;

y is a number from 1 to 20, preferably from 2 to 10, more preferably from 2 to 8, more preferably from 2 to 6 or 4 to 8, and most preferably from 2, 4, 6, 7, 8;

Ab is an anti-HER3 antibody or an antigen-binding fragment thereof, which comprises: an antibody heavy chain variable region comprising HCDR1, HCDR2 and HCDR3 regions and an antibody light chain variable region comprising LCDR1, LCDR2 and LCDR3 regions, wherein,

The antibody heavy chain variable region of the Ab includes:

HCDR1 as shown in SEQ ID NO: 08,

HCDR2 as shown in SEQ ID NO: 27, and

HCDR3 as shown in SEQ ID NO: 46;

or

HCDR1 as shown in SEQ ID NO: 09,

HCDR2 as shown in SEQ ID NO: 28, and

HCDR3 as shown in SEQ ID NO: 47;

or

HCDR1 as shown in SEQ ID NO: 10,

HCDR2 as shown in SEQ ID NO: 29, and

HCDR3 as shown in SEQ ID NO: 48;

or

HCDR1 as shown in SEQ ID NO: 11,

HCDR2 as shown in SEQ ID NO: 30, and

HCDR3 as shown in SEQ ID NO: 49;

or

HCDR1 as shown in SEQ ID NO: 12,

HCDR2 as shown in SEQ ID NO: 31, and

HCDR3 as shown in SEQ ID NO: 50;

or

HCDR1 as shown in SEQ ID NO: 01,

HCDR2 as shown in SEQ ID NO: 21, and

HCDR3 as shown in SEQ ID NO: 51;

or

HCDR1 as shown in SEQ ID NO: 06,

HCDR2 as shown in SEQ ID NO: 32, and

HCDR3 as shown in SEQ ID NO: 52;

or

HCDR1 as shown in SEQ ID NO: 13,

HCDR2 as shown in SEQ ID NO: 33, and

HCDR3 as shown in SEQ ID NO: 53;

or

HCDR1 as shown in SEQ ID NO: 14,

HCDR2 as shown in SEQ ID NO: 33, and

HCDR3 as shown in SEQ ID NO: 54;

or

HCDR1 as shown in SEQ ID NO: 15,

HCDR2 as shown in SEQ ID NO: 34, and

HCDR3 as shown in SEQ ID NO: 55;

or

HCDR1 as shown in SEQ ID NO: 16,

HCDR2 as shown in SEQ ID NO: 35, and

HCDR3 as shown in SEQ ID NO: 56;

or

HCDR1 as shown in SEQ ID NO: 17,

HCDR2 as shown in SEQ ID NO: 36, and

HCDR3 as shown in SEQ ID NO: 57;

or

HCDR1 as shown in SEQ ID NO: 06,

HCDR2 as shown in SEQ ID NO: 32, and

HCDR3 as shown in SEQ ID NO: 58;

or

HCDR1 as shown in SEQ ID NO: 01,

HCDR2 as shown in SEQ ID NO: 18, and

HCDR3 as shown in SEQ ID NO: 37;

or

HCDR1 as shown in SEQ ID NO: 02,

HCDR2 as shown in SEQ ID NO: 19, and

HCDR3 as shown in SEQ ID NO: 38;

or

HCDR1 as shown in SEQ ID NO: 03,

HCDR2 as shown in SEQ ID NO: 20, and

HCDR3 as shown in SEQ ID NO: 39;

or

HCDR1 as shown in SEQ ID NO: 04,

HCDR2 as shown in SEQ ID NO: 21, and

HCDR3 as shown in SEQ ID NO: 40;

or

HCDR1 as shown in SEQ ID NO: 05,

HCDR2 as shown in SEQ ID NO: 22, and

HCDR3 as shown in SEQ ID NO: 41;

or

HCDR1 as shown in SEQ ID NO: 06,

HCDR2 as shown in SEQ ID NO: 23, and

HCDR3 as shown in SEQ ID NO: 42;

or

HCDR1 as shown in SEQ ID NO: 07,

HCDR2 as shown in SEQ ID NO: 24, and

HCDR3 as shown in SEQ ID NO: 43;

or

HCDR1 as shown in SEQ ID NO: 06,

HCDR2 as shown in SEQ ID NO: 25, and

HCDR3 as shown in SEQ ID NO: 44;

or

HCDR1 as shown in SEQ ID NO: 06,

HCDR2 as shown in SEQ ID NO: 26, and

HCDR3 as shown in SEQ ID NO: 45;

and/or

The antibody light chain variable region of the Ab comprises:

LCDR1 as shown in SEQ ID NO: 66,

LCDR2 as shown in SEQ ID NO: 83, and

LCDR3 as shown in SEQ ID NO: 96;

or

LCDR1 as shown in SEQ ID NO: 67,

LCDR2 as shown in SEQ ID NO: 79, and

LCDR3 as shown in SEQ ID NO: 97;

or

LCDR1 as shown in SEQ ID NO: 68,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 98;

or

LCDR1 as shown in SEQ ID NO: 69,

LCDR2 as shown in SEQ ID NO: 84, and

LCDR3 as shown in SEQ ID NO: 99;

or

LCDR1 as shown in SEQ ID NO: 70,

LCDR2 as shown in SEQ ID NO: 85, and

LCDR3 as shown in SEQ ID NO: 100;

or

LCDR1 as shown in SEQ ID NO: 71,

LCDR2 as shown in SEQ ID NO: 85, and

LCDR3 as shown in SEQ ID NO: 100;

or

LCDR1 as shown in SEQ ID NO: 61,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 90;

or

LCDR1 as shown in SEQ ID NO: 72,

LCDR2 as shown in SEQ ID NO: 86, and

LCDR3 as shown in SEQ ID NO: 101;

or

LCDR1 as shown in SEQ ID NO: 73,

LCDR2 as shown in SEQ ID NO: 82, and

LCDR3 as shown in SEQ ID NO: 103;

or

LCDR1 as shown in SEQ ID NO: 64,

LCDR2 as shown in SEQ ID NO: 82, and

LCDR3 as shown in SEQ ID NO: 103;

or

LCDR1 as shown in SEQ ID NO: 74,

LCDR2 as shown in SEQ ID NO: 79, and

LCDR3 as shown in SEQ ID NO: 92;

or

LCDR1 as shown in SEQ ID NO: 75,

LCDR2 as shown in SEQ ID NO: 82, and

LCDR3 as shown in SEQ ID NO: 104;

or

LCDR1 as shown in SEQ ID NO: 65,

LCDR2 as shown in SEQ ID NO: 82, and

LCDR3 as shown in SEQ ID NO: 95;

or

LCDR1 as shown in SEQ ID NO: 76,

LCDR2 as shown in SEQ ID NO: 85, and

LCDR3 as shown in SEQ ID NO: 100;

or

LCDR1 as shown in SEQ ID NO: 66,

LCDR2 as shown in SEQ ID NO: 83, and

LCDR3 as shown in SEQ ID NO: 96;

or

LCDR1 as shown in SEQ ID NO: 59,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 58;

or

LCDR1 as shown in SEQ ID NO: 60,

LCDR2 as shown in SEQ ID NO: 78, and

LCDR3 as shown in SEQ ID NO: 89;

or

LCDR1 as shown in SEQ ID NO: 61,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 90;

or

LCDR1 as shown in SEQ ID NO: 59,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 90;

or

LCDR1 as shown in SEQ ID NO: 62,

LCDR2 as shown in SEQ ID NO: 77, and

LCDR3 as shown in SEQ ID NO: 91;

or

LCDR1 as shown in SEQ ID NO: 63,

LCDR2 as shown in SEQ ID NO: 79, and

LCDR3 as shown in SEQ ID NO: 92;

or

LCDR1 as shown in SEQ ID NO: 60,

LCDR2 as shown in SEQ ID NO: 80, and

LCDR3 as shown in SEQ ID NO: 93;

or

LCDR1 as shown in SEQ ID NO: 64,

LCDR2 as shown in SEQ ID NO: 81, and

LCDR3 as shown in SEQ ID NO: 94;

or

LCDR1 as shown in SEQ ID NO: 65,

LCDR2 as shown in SEQ ID NO: 82, and

LCDR3 as shown in SEQ ID NO: 95;

or

LCDR1 as shown in SEQ ID NO: 60,

LCDR2 as shown in SEQ ID NO: 80, and

LCDR3 as shown in SEQ ID NO: 89.

The antibody-drug conjugate or its pharmaceutically acceptable salt or solvate as described in claim 1, wherein Ab comprises:

a) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 08, SEQ ID NO: 27 and SEQ ID NO: 46, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 66, SEQ ID NO: 83 and SEQ ID NO: 96, respectively;

or

b) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 09, SEQ ID NO: 28 and SEQ ID NO: 47, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 67, SEQ ID NO: 79 and SEQ ID NO: 97, respectively;

or

c) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 10, SEQ ID NO: 29 and SEQ ID NO: 48, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 68, SEQ ID NO: 77 and SEQ ID NO: 98, respectively;

or

d) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 11, SEQ ID NO: 30 and SEQ ID NO: 49, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 69, SEQ ID NO: 84 and SEQ ID NO: 99, respectively;

or

e) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 12, SEQ ID NO: 31 and SEQ ID NO: 50, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 70, SEQ ID NO: 85 and SEQ ID NO: 100, respectively;

or

f) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 12, SEQ ID NO: 31 and SEQ ID NO: 50, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 71, SEQ ID NO: 85 and SEQ ID NO: 100, respectively;

or

g) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 01, SEQ ID NO: 21 and SEQ ID NO: 51, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 61, SEQ ID NO: 77 and SEQ ID NO: 90, respectively;

or

h) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 52, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 72, SEQ ID NO: 86 and SEQ ID NO: 101, respectively;

or

i) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 52, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 73, SEQ ID NO: 87 and SEQ ID NO: 102, respectively;

or

j) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 13, SEQ ID NO: 33 and SEQ ID NO: 53, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 73, SEQ ID NO: 82 and SEQ ID NO: 103, respectively;

or

k) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 14, SEQ ID NO: 33 and SEQ ID NO: 54, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 64, SEQ ID NO: 82 and SEQ ID NO: 103, respectively;

or

l) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 15, SEQ ID NO: 34 and SEQ ID NO: 55, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 74, SEQ ID NO: 79 and SEQ ID NO: 92, respectively;

or

m) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 16, SEQ ID NO: 35 and SEQ ID NO: 56, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 75, SEQ ID NO: 82 and SEQ ID NO: 104, respectively;

or

n) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 16, SEQ ID NO: 35 and SEQ ID NO: 56, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 65, SEQ ID NO: 82 and SEQ ID NO: 95, respectively;

or

o) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 17, SEQ ID NO: 36 and SEQ ID NO: 57, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 76, SEQ ID NO: 85 and SEQ ID NO: 100, respectively;

or

p) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 32 and SEQ ID NO: 58, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 66, SEQ ID NO: 83 and SEQ ID NO: 96, respectively;

or

q) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 01, SEQ ID NO: 18 and SEQ ID NO: 37, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 59, SEQ ID NO: 77 and SEQ ID NO: 88, respectively;

or

r) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 02, SEQ ID NO: 19 and SEQ ID NO: 38, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 60, SEQ ID NO: 78 and SEQ ID NO: 89, respectively;

or

s) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 03, SEQ ID NO: 20 and SEQ ID NO: 39, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 61, SEQ ID NO: 77 and SEQ ID NO: 90, respectively;

or

t) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 04, SEQ ID NO: 21 and SEQ ID NO: 40, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 59, SEQ ID NO: 77 and SEQ ID NO: 90, respectively;

or

u) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 05, SEQ ID NO: 22 and SEQ ID NO: 41, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 62, SEQ ID NO: 77 and SEQ ID NO: 91, respectively;

or

v) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 23 and SEQ ID NO: 42, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 63, SEQ ID NO: 79 and SEQ ID NO: 92, respectively;

or

w) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 07, SEQ ID NO: 24 and SEQ ID NO: 43, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 60, SEQ ID NO: 80 and SEQ ID NO: 93, respectively;

or

x) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 25 and SEQ ID NO: 44, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 64, SEQ ID NO: 81 and SEQ ID NO: 94, respectively;

or

y) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 06, SEQ ID NO: 26 and SEQ ID NO: 45, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 65, SEQ ID NO: 82 and SEQ ID NO: 95, respectively;

or

z) The heavy chain variable region sequence comprises HCDR1, HCDR2 and HCDR3 as shown in SEQ ID NO: 08, SEQ ID NO: 27 and SEQ ID NO: 46, respectively; and the light chain variable region sequence comprises LCDR1, LCDR2 and LCDR3 as shown in SEQ ID NO: 60, SEQ ID NO: 80 and SEQ ID NO: 89, respectively.

The antibody-drug conjugate or its pharmaceutically acceptable salt or solvate as described in claim 1 or 2, wherein the Ab is a mouse antibody, a chimeric antibody, a humanized antibody, a human antibody or an antigen-binding fragment thereof.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 3, wherein the Ab comprises a heavy chain variable region of the following sequence: SEQ ID NO: 127, 129, 131, 133, 135, 137, 139, 141, 144, 145, 147, 149, 151, 153, 155, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123 and 125, or having at least 80%, 85%, 90%, 95% or 99% sequence identity therewith;

and/or

The Ab comprises a light chain variable region of the following sequence: SEQ ID NO: 128, 130, 132, 134, 136, 138, 140, 142, 143, 146, 148, 150, 152, 154, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124 and 126, or has at least 80%, 85%, 90%, 95% or 99% sequence identity thereto.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 4, wherein the Ab comprises:

a-1) a heavy chain variable region as shown in SEQ ID NO: 127 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 128 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

a-2) a heavy chain variable region as shown in SEQ ID NO: 127 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 126 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

b) a heavy chain variable region as shown in SEQ ID NO: 129 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 130 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

c) a heavy chain variable region as shown in SEQ ID NO: 131 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 132 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

d) a heavy chain variable region as shown in SEQ ID NO: 133 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 134 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

e-1) a heavy chain variable region as shown in SEQ ID NO: 135 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 136 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

e-2) a heavy chain variable region as shown in SEQ ID NO: 137 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 138 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

f) a heavy chain variable region as shown in SEQ ID NO: 139 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 140 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

g-1) a heavy chain variable region as shown in SEQ ID NO: 141 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 142 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

g-2) a heavy chain variable region as shown in SEQ ID NO: 141 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 143 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

g-3) a heavy chain variable region as shown in SEQ ID NO: 144 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 142 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

h) a heavy chain variable region as shown in SEQ ID NO: 145 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 146 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

i) a heavy chain variable region as shown in SEQ ID NO: 147 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 148 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

j) a heavy chain variable region as shown in SEQ ID NO: 149 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 150 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

k-1) a heavy chain variable region as shown in SEQ ID NO: 151 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 152 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

k-2) a heavy chain variable region as shown in SEQ ID NO: 151 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 122 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

l) a heavy chain variable region as shown in SEQ ID NO: 153 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 154 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

m) a heavy chain variable region as shown in SEQ ID NO: 155 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 126 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

n) a heavy chain variable region as shown in SEQ ID NO: 105 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 106 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

o) a heavy chain variable region as shown in SEQ ID NO: 107 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 108 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

p) a heavy chain variable region as shown in SEQ ID NO: 109 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 110 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

q) a heavy chain variable region as shown in SEQ ID NO: 111 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 112 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

r) a heavy chain variable region as shown in SEQ ID NO: 113 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 114 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

s) a heavy chain variable region as shown in SEQ ID NO: 115 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 116 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

t) a heavy chain variable region as shown in SEQ ID NO: 117 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 118 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

u) a heavy chain variable region as shown in SEQ ID NO: 119 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 120 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

v) a heavy chain variable region as shown in SEQ ID NO: 121 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 122 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

w-1) a heavy chain variable region as shown in SEQ ID NO: 123 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 124 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

w-2) a heavy chain variable region as shown in SEQ ID NO: 125 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 126 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto;

or

x) a heavy chain variable region as shown in SEQ ID NO: 123 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto; and/or a light chain variable region as shown in SEQ ID NO: 126 or having at least 80%, 85%, 90%, 95% or 99% sequence identity thereto.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 5, wherein the Ab comprises:

a-1) a heavy chain variable region as shown in SEQ ID NO: 127 and a light chain variable region as shown in SEQ ID NO: 128;

a-2) a heavy chain variable region as shown in SEQ ID NO: 127 and a light chain variable region as shown in SEQ ID NO: 126;

b) a heavy chain variable region as shown in SEQ ID NO: 129 and a light chain variable region as shown in SEQ ID NO: 130;

c) a heavy chain variable region as shown in SEQ ID NO: 131 and a light chain variable region as shown in SEQ ID NO: 132;

d) a heavy chain variable region as shown in SEQ ID NO: 133 and a light chain variable region as shown in SEQ ID NO: 134;

e-1) a heavy chain variable region as shown in SEQ ID NO: 135 and a light chain variable region as shown in SEQ ID NO: 136;

e-2) a heavy chain variable region as shown in SEQ ID NO: 137 and a light chain variable region as shown in SEQ ID NO: 138;

f) a heavy chain variable region as shown in SEQ ID NO: 139 and a light chain variable region as shown in SEQ ID NO: 140;

g-1) a heavy chain variable region as shown in SEQ ID NO: 141 and a light chain variable region as shown in SEQ ID NO: 142;

g-2) a heavy chain variable region as shown in SEQ ID NO: 141 and a light chain variable region as shown in SEQ ID NO: 143;

g-3) a heavy chain variable region as shown in SEQ ID NO: 144 and a light chain variable region as shown in SEQ ID NO: 142;

h) a heavy chain variable region as shown in SEQ ID NO: 145 and a light chain variable region as shown in SEQ ID NO: 146;

i) a heavy chain variable region as shown in SEQ ID NO: 147 and a light chain variable region as shown in SEQ ID NO: 148;

j) a heavy chain variable region as shown in SEQ ID NO: 149 and a light chain variable region as shown in SEQ ID NO: 150;

k-1) a heavy chain variable region as shown in SEQ ID NO: 151 and a light chain variable region as shown in SEQ ID NO: 152;

k-2) a heavy chain variable region as shown in SEQ ID NO: 151 and a light chain variable region as shown in SEQ ID NO: 122;

l) a heavy chain variable region as shown in SEQ ID NO: 153 and a light chain variable region as shown in SEQ ID NO: 154;

m) a heavy chain variable region as shown in SEQ ID NO: 155 and a light chain variable region as shown in SEQ ID NO: 126;

n) a heavy chain variable region as shown in SEQ ID NO: 105 and a light chain variable region as shown in SEQ ID NO: 106;

o) a heavy chain variable region as shown in SEQ ID NO: 107 and a light chain variable region as shown in SEQ ID NO: 108;

p) a heavy chain variable region as shown in SEQ ID NO: 109 and a light chain variable region as shown in SEQ ID NO: 110;

q) a heavy chain variable region as shown in SEQ ID NO: 111 and a light chain variable region as shown in SEQ ID NO: 112;

r) a heavy chain variable region as shown in SEQ ID NO: 113 and a light chain variable region as shown in SEQ ID NO: 114;

s) a heavy chain variable region as shown in SEQ ID NO: 115 and a light chain variable region as shown in SEQ ID NO: 116;

t) a heavy chain variable region as shown in SEQ ID NO: 117 and a light chain variable region as shown in SEQ ID NO: 118;

u) a heavy chain variable region as shown in SEQ ID NO: 119 and a light chain variable region as shown in SEQ ID NO: 120;

v) a heavy chain variable region as shown in SEQ ID NO: 121 and a light chain variable region as shown in SEQ ID NO: 122;

w-1) a heavy chain variable region as shown in SEQ ID NO: 123 and a light chain variable region as shown in SEQ ID NO: 124;

w-2) a heavy chain variable region as shown in SEQ ID NO: 125 and a light chain variable region as shown in SEQ ID NO: 126;

x) a heavy chain variable region as shown in SEQ ID NO: 123 and a light chain variable region as shown in SEQ ID NO: 126.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 6, wherein the Ab is a full-length antibody, which further comprises a human antibody constant region;

Preferably, the heavy chain constant region of the human antibody constant region is selected from the constant regions of human IgG1, IgG2, IgG3 and IgG4 and conservative variants thereof, and the light chain constant region of the human antibody constant region is selected from the kappa chain constant region and the lambda chain constant region of human antibodies and conservative variants thereof;

More preferably, the full-length antibody comprises the human antibody heavy chain constant region of SEQ ID NO: 156 and the human light chain constant region of SEQ ID NO: 157.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 7, wherein the Ab comprises:

a-1) a heavy chain as shown in SEQ ID NO: 180 and a light chain as shown in SEQ ID NO: 181;

a-2) a heavy chain as shown in SEQ ID NO: 180 and a light chain as shown in SEQ ID NO: 179;

b) a heavy chain as shown in SEQ ID NO: 182 and a light chain as shown in SEQ ID NO: 183;

c) a heavy chain as shown in SEQ ID NO: 184 and a light chain as shown in SEQ ID NO: 185;

g) a heavy chain as shown in SEQ ID NO: 186 and a light chain as shown in SEQ ID NO: 187;

e-1) a heavy chain as shown in SEQ ID NO: 188 and a light chain as shown in SEQ ID NO: 189;

e-2) a heavy chain as shown in SEQ ID NO: 190 and a light chain as shown in SEQ ID NO: 191;

f) a heavy chain as shown in SEQ ID NO: 192 and a light chain as shown in SEQ ID NO: 193;

g-1) a heavy chain as shown in SEQ ID NO: 194 and a light chain as shown in SEQ ID NO: 195;

g-2) a heavy chain as shown in SEQ ID NO: 194 and a light chain as shown in SEQ ID NO: 196;

g-3) a heavy chain as shown in SEQ ID NO: 197 and a light chain as shown in SEQ ID NO: 195;

h) a heavy chain as shown in SEQ ID NO: 198 and a light chain as shown in SEQ ID NO: 199;

i) a heavy chain as shown in SEQ ID NO: 200 and a light chain as shown in SEQ ID NO: 201;

j) a heavy chain as shown in SEQ ID NO: 202 and a light chain as shown in SEQ ID NO: 203;

k-1) a heavy chain as shown in SEQ ID NO: 204 and a light chain as shown in SEQ ID NO: 205;

k-2) a heavy chain as shown in SEQ ID NO: 204 and a light chain as shown in SEQ ID NO: 175;

l) a heavy chain as shown in SEQ ID NO: 206 and a light chain as shown in SEQ ID NO: 207;

m) a heavy chain as shown in SEQ ID NO: 208 and a light chain as shown in SEQ ID NO: 179;

n) a heavy chain as shown in SEQ ID NO: 158 and a light chain as shown in SEQ ID NO: 159;

o) a heavy chain as shown in SEQ ID NO: 160 and a light chain as shown in SEQ ID NO: 161;

p) a heavy chain as shown in SEQ ID NO: 162 and a light chain as shown in SEQ ID NO: 163;

q) a heavy chain as shown in SEQ ID NO: 164 and a light chain as shown in SEQ ID NO: 165;

r) a heavy chain as shown in SEQ ID NO: 166 and a light chain as shown in SEQ ID NO: 167;

s) a heavy chain as shown in SEQ ID NO: 168 and a light chain as shown in SEQ ID NO: 169;

t) a heavy chain as shown in SEQ ID NO: 170 and a light chain as shown in SEQ ID NO: 171;

u) a heavy chain as shown in SEQ ID NO: 172 and a light chain as shown in SEQ ID NO: 173;

v) a heavy chain as shown in SEQ ID NO: 174 and a light chain as shown in SEQ ID NO: 175;

w-1) a heavy chain as shown in SEQ ID NO: 176 and a light chain as shown in SEQ ID NO: 177;

w-2) a heavy chain as shown in SEQ ID NO: 178 and a light chain as shown in SEQ ID NO: 179;

x) a heavy chain as shown in SEQ ID NO: 176 and a light chain as shown in SEQ ID NO: 179.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 8, wherein the antigen-binding fragment is selected from: Fab, Fab', F(ab')2, variable fragment (Fv), single-chain variable fragment (scFv), dimerization domain V (diabody), disulfide-stabilized Fv (dsFv) and CDR-containing peptide.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 9, wherein the cytotoxic drug is selected from toxins, chemotherapeutic agents, antibiotics, radioactive isotopes and nucleolytic enzymes.

The antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 10, wherein the cytotoxic drug is selected from tubulin inhibitors or topoisomerase inhibitors; preferably auristatin analogs or camptothecin derivatives; more preferably SN-38, MMAE, MMAF or exotecan.

The antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in claim 11, wherein the antibody-drug conjugate is represented by the general formula (B):

in,

_L1 and _L2 are connection units;

y is a number selected from 1 to 10, preferably a number selected from 2 to 8, more preferably a number from 2 to 6 or 4 to 8, further preferably a number from 6 to 8, and most preferably 4, 6, 7, 8;

Ab is an anti-HER3 antibody or an antigen-binding fragment thereof as described in any one of claims 1 to 9.

The antibody-drug conjugate or its pharmaceutically acceptable salt or solvate as described in claim 12, wherein the antibody-drug conjugate is represented by the general formula (C):

in,

_R1 is selected from hydrogen, _C1-6 halogen alkyl or _C3-8 cycloalkyl;

_R2 is selected from hydrogen, _C1-6 halogen alkyl or _C3-8 cycloalkyl;

Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-8 cycloalkyl group.

The antibody-drug conjugate or its pharmaceutically acceptable salt or solvate as described in claim 13, wherein:

_R1 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

_R2 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-6 cycloalkyl group.

The antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 12 to 14, wherein _L2 is represented by the general formula (I):

in,

^s1 and ^s2 are each independently an integer selected from 0-8, preferably, s1 and ^s2 are independently selected from 1, ² , 3, 4, 5 or 6;

or, ^s1 is an integer from 1 to 8, ^s2 is 0, preferably, ^s1 is selected from 4, 5, 6, 7 or 8, and ^s2 is 0;

Or, ^s2 is selected from an integer from 2 to 8, ^s1 is 2, preferably, ^s2 is selected from 2, 3, 4, 5 or 6, and ^s1 is 2.

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 12 to 15, wherein the antibody-drug conjugate has the following structure:

in,

An antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof as described in any one of claims 1 to 16, wherein the antibody-drug conjugate is selected from the following compounds:

in,

y is a number selected from 1 to 10, preferably a number selected from 2 to 8, more preferably a number from 2 to 6 or 4 to 8, further preferably a number from 6 to 8, and most preferably 4, 6, 7, 8.

A method for preparing an antibody-drug conjugate or a pharmaceutically acceptable salt or solvate thereof, the method comprising the following steps:

After reduction, Ab is coupled to the compound to obtain an antibody-drug conjugate;

in,

Ab is an anti-HER3 antibody or an antigen-binding fragment thereof as described in any one of claims 1 to 9;

_R1 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

_R2 is selected from hydrogen, _C1-3 halogen alkyl or _C3-6 cycloalkyl;

Or, _R1 and _R2 together with the carbon atom to which they are attached form a _C3-6 cycloalkyl group;

_L2 is a connector as described in claim 15;

A pharmaceutical composition comprising an antibody-drug conjugate as described in any one of claims 1 to 17 or a pharmaceutically acceptable salt or solvate thereof, and one or more pharmaceutically acceptable excipients, diluents or carriers.

An antibody-drug conjugate as described in any one of claims 1 to 17 or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition as described in claim 19, for use in the manufacture of a drug for treating or preventing a disease associated with human HER3.

The use as described in claim 20, wherein it is used to manufacture a method for treating or preventing cancer expressing HER3; preferably, the cancer is breast cancer, colorectal cancer, lung cancer, multiple myeloma, ovarian cancer, liver cancer, gastric cancer, pancreatic cancer, prostate cancer, acute myeloid leukemia, chronic myeloid leukemia, osteosarcoma, squamous cell carcinoma, peripheral neurilemmoma, neurothecoma, head and neck cancer, bladder cancer, esophageal cancer, glioblastoma, soft tissue clear cell sarcoma, malignant mesothelioma, neurofibromatosis, kidney cancer and melanoma.