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抗體藥物偶聯(lián)物:全面指南

抗體藥物偶聯(lián)物(ADCs)是化療和免疫療法的一種強大組合。這個(gè)概念最早由德國科學(xué)家保羅·埃爾利希在100多年前提出。他將ADCs比作“魔法子彈”,因為它們可以特異性地識別目標(癌細胞),而不會(huì )對機體造成傷害,就像“狙擊手”一樣。近年來(lái),隨著(zhù)越來(lái)越多的ADCs獲得批準并上市,這種藥物已成為當前全球生物技術(shù)藥物研發(fā)中最熱門(mén)的技術(shù)領(lǐng)域之一。那么,ADCs為什么如此令人驚奇?它們是如何實(shí)現“精準引導”的?本文收集了ADCs的一些常見(jiàn)問(wèn)題,包括:


1. 什么是抗體藥物偶聯(lián)物?

抗體藥物偶聯(lián)物(ADCs)是一種新型生物藥物,它將單克隆抗體藥物的高特異性與小分子細胞毒藥物的高活性結合在一起,以提高腫瘤藥物的靶向性并減少毒性和副作用。


2. 抗體藥物偶聯(lián)物的結構

ADCs由三個(gè)主要部分組成:抗體負責選擇性地識別癌細胞表面抗原,藥物負責殺死癌細胞,連接劑用于將抗體與藥物負載物連接在一起。

ADCs的結構

圖1. ADCs的結構 [1]


3. 抗體藥物偶聯(lián)物是如何起作用的?

首先,將ADCs注射到體內后,經(jīng)過(guò)許多障礙物后,它們會(huì )與目標細胞的抗原結合,形成ADCs-抗原復合物,并通過(guò)網(wǎng)格介導的內吞作用進(jìn)入細胞。復合物進(jìn)入細胞后,溶酶體與內體融合導致細胞內連接劑的裂解,釋放和激活小分子細胞毒藥物。藥物釋放到細胞質(zhì)后,可以插入DNA或抑制微管聚合,殺死腫瘤細胞并導致靶細胞凋亡。當目標細胞死亡時(shí),活性負荷還可能殺死周?chē)哪[瘤細胞,這也被稱(chēng)為旁觀(guān)效應。

ADCs的工作機制

圖1. ADCs的工作機制 [1]


4. 抗體藥物偶聯(lián)物的組成成分

4.1 靶點(diǎn)的選擇

ADCs的成功開(kāi)發(fā)取決于抗體對靶點(diǎn)的特異性結合。理想的ADCs靶點(diǎn)應在腫瘤細胞表面高表達,在正常細胞中低表達或不表達,并盡量減少對目標細胞和非腫瘤組織的毒性。除了特異性和足夠的表達量外,最好的靶點(diǎn)還應該引起高效的內吞作用效應。

Target antigens in approved ADCs and selected
ADCs in late-stage clinical development

Target antigens regulated from
driver oncogenes

Target antigens in the tumor
vasculature and stroma

圖3. 正在開(kāi)發(fā)和臨床開(kāi)發(fā)的adc靶點(diǎn)

4.2 抗體的選擇

抗體應具有高的抗原親和力和長(cháng)的循環(huán)半衰期,以便在腫瘤部位特異性富集細胞毒素??紤]到不同IgG類(lèi)型抗體的半衰期、結構穩定性、Fc段的免疫功能和偶聯(lián)方便性,ADCs主要選擇IgG1,但很少使用IgG4。根據抗體的免疫原性,它們可分為全人源化抗體、人源化抗體和嵌合抗體。為了減少免疫反應,通常選擇完全人源化或人源化抗體。

4.3 藥物負荷的選擇

毒素分子(藥物負荷)是ADCs研究和開(kāi)發(fā)成功的關(guān)鍵因素。由于A(yíng)DCs從進(jìn)入人體到最終釋放細胞毒素需要經(jīng)歷多個(gè)步驟,考慮到每個(gè)步驟的效率,藥物負荷應具有高的抗腫瘤活性,因此納摩爾水平(IC50值為0.01-0.1 nM)的毒性分子是合適的藥物負荷。此外,藥物負荷還必須具有適當的可偶聯(lián)的功能基團、強大的細胞毒性、適當的親水性-疏水性平衡以及高穩定性。

4.4 連接劑的選擇

連接劑作為ADCs的橋梁,通過(guò)可裂解連接劑或不可裂解連接劑與抗體連接。連接劑需要精心設計,既需要在生理狀態(tài)下具有穩定性,以防止鏈斷裂,又需要在特定部位具有高釋放效率。


5. 抗體藥物偶聯(lián)物的優(yōu)勢

與傳統的完全或部分人源化抗體或抗體片段相比,ADCs具有更高的理論治療效果,因為它們可以在腫瘤組織中釋放高活性的細胞毒素。另一方面,ADCs的耐受性更高,副作用較低,與融合蛋白相比。它們可以精確識別目標并不影響正常細胞,大大提高了療效并減少了毒性和副作用,因此在藥物研發(fā)領(lǐng)域引起了人們的關(guān)注。


6. 抗體藥物偶聯(lián)物有哪些弱點(diǎn)?

ADCs的結構復雜,設計多樣,這給與CMC研究相關(guān)的生產(chǎn)和質(zhì)量控制帶來(lái)了困難。同時(shí),ADCs與體內復雜的生物過(guò)程疊加,也使非臨床研究和臨床研究面臨多重挑戰。由于A(yíng)DCs的生產(chǎn)大多涉及高活性細胞毒性藥物,因此需要高度的硬件設備、流程設計和人員培訓,并需要大量的資本投入和技術(shù)儲備,因此生產(chǎn)門(mén)檻較高。

目前,抗體藥物偶聯(lián)物的發(fā)展面臨三個(gè)主要挑戰和機遇:

6.1 連接劑的不穩定性

這種不穩定性可能導致藥物提前釋放到血液中,導致ADCs的非特異性攝取和錯靶毒性。

6.2 非特異性?xún)韧套饔?/h3>

親水性將促進(jìn)ADCs的聚集和非特異性?xún)韧套饔?,尤其是DAR(藥物-抗體比)較高的ADCs,導致錯靶效應。DAR較高的ADCs還會(huì )被其他具有非特異性和強大內吞作用能力的細胞清除。因此,優(yōu)化DAR也是改善治療指數(TI)的重要策略。

6.3 受體介導的攝取機制

由Fcγ受體介導的ADC的錯靶毒性主要表現為血液毒性。血液毒性是含有Auristatin(MAME、MMAF)、Calicheamicin和Maytansinoid(DM-1)的ADCs最常見(jiàn)的非特異性錯靶劑量限制毒性(DLTs)。


7. 獲得FDA批準的抗體藥物偶聯(lián)物

在獲得批準上市的十種ADCs中,有六種用于治療血液腫瘤,其他的用于固體腫瘤(表1)。目前,有超過(guò)80種ADCs正在進(jìn)行積極的臨床試驗,其中大部分處于I期和I/II期。超過(guò)80%的臨床試驗研究ADCs在各種固體腫瘤中的安全性和有效性,而其余涉及血液惡性腫瘤。這表明,在早期T-DM1的成功和最近對sacituzumab govitecan和Loncastuximab tesirine的批準后,近年來(lái)ADCs的研究逐漸轉向固體腫瘤。

表1.FDA批準的ADC

藥物名稱(chēng) 商品名 公司 靶點(diǎn) 有效負載 適應癥 審批年度
瓊妥珠單抗奧唑加霉素 Mylotarg 輝瑞/惠氏 CD33 Calicheamicin AML 2000
2017
布倫妥昔單抗 Adcetris 西雅圖遺傳學(xué)公司, 武田腫瘤 CD30 MMAE cHL、sALCL、PTCL 2011
曲妥珠單抗 Kadcyla 基因泰克/羅氏 HER2 DM1 mBC 2013
伊諾妥珠單抗奧唑加霉素 Besponsa 輝瑞/惠氏 CD22 Calicheamicin ALL 2017
泊洛妥珠單抗 Polivy 基因泰克/羅氏 CD79b MMAE r/r DLBCL 2019
恩諾單抗 Padcev 安斯泰來(lái)/西雅圖遺傳學(xué)公司 Nectin-4 MMAE mUC 2019
曲妥珠單抗 Enhertu 阿斯利康/ 第一三共株式會(huì )社 HER2 Dxd mBC、mGC 2019
戈沙妥珠單抗 Trodelvy Immunomedics TROP2 SN-38 mTNBC 2020
馬貝蘭他單抗 Blenrep 葛蘭素史克 BCMA MMAF MM 2020
朗妥昔單抗 Zynlonta ADC療法 CD19 SG3199 r/r DLBCL 2021

自從“生物導彈”的概念提出以來(lái),ADCs一直在不斷創(chuàng )新和優(yōu)化,并得到了極大的改進(jìn),使其成為癌癥治療領(lǐng)域的重要手段之一。隨著(zhù)越來(lái)越多的ADCs進(jìn)入臨床階段,該行業(yè)正逐漸從傳統技術(shù)轉向更具創(chuàng )新性的技術(shù)來(lái)開(kāi)發(fā)這種復雜產(chǎn)品,其中包括探索新的腫瘤抗原、新的抗體結構、新的藥物負荷、新的連接劑和先進(jìn)的偶聯(lián)方法。隨著(zhù)對ADCs的深入研究,ADCs的分子結構設計將更加合理,均一性將大大提高,體內穩定性將不斷改善,從而減少毒性和副作用,提高療效和活性,擴大治療窗口,為腫瘤患者帶來(lái)新的希望。


8. 華美生物ADC靶點(diǎn)蛋白

目前,ADC在研靶點(diǎn)有128個(gè),其中有2條及以上在研管線(xiàn)的59個(gè)。華美生物已經(jīng)針對各種熱門(mén)靶點(diǎn)開(kāi)發(fā)了一系列具有不同物種和標簽的產(chǎn)品,適用于免疫、抗體篩選、SPR、細胞活性檢測等實(shí)驗。華美生物致力于協(xié)助您的藥物研發(fā)工作,并且所有活性測試方案都可以免費獲取。此外,華美生物還提供相關(guān)的研究用抗體和ELISA試劑盒。如果您沒(méi)有找到所需的靶點(diǎn),您可以通過(guò)留言或在線(xiàn)聊天與我們聯(lián)系。

● 部分ADC靶點(diǎn)蛋白活性驗證數據

TACSTD2 (TROP2)
CSB-MP023072HU1
CSB-MP023072HU1 Activity Verified

Measured in cell activity assay using U937 cells, the EC50 for this effect is 190.2-298.6 ng/ml.

CSB-MP007479HU Activity Verified

Human EGF protein captured on COOH chip can bind Human EGFR protein, his and Myc tag (CSB-MP007479HU) with an affinity constant of 11.9nM as detected by LSPR Assay.

CSB-MP013714HU Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized MET at 2 μg/ml can bind Anti-MET recombinant antibody, the EC50 is 2.379-3.094 ng/ml.

CSB-MP024093HUb0 Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized Human TPBG at 2 μg/mL can bind Anti-TPBG recombinant antibody (CSB-RA024093MA1HU), the EC50 is 1.230-1.519 ng/mL.

CSB-MP822274HU Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized NECTIN4 at 2 μg/ml can bind anti-NECTIN4 antibody (CSB-RA822274A0HU) (enfortumab vedotin-like), the EC50 is 0.6029-0.7837 ng/mL.

CSB-MP733578HU Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized CD276 at 2 μg/ml can bind Anti-CD276 rabbit monoclonal antibody, the EC50 of human CD276 protein is 1.961-2.243 ng/ml.

CSB-MP015044HUc9 Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized MSLN at 2 μg/ml can bind Anti-MSLN rabbit monoclonal antibody, the EC50 of the MSLN protein is 2.657-3.177 ng/ml.

CSB-MP004954HU1 Activity Verified

Measured by its binding ability in a functional ELISA. Immobilized Human CD70 at 2 μg/ml can bind Anti-CD70 antibody, the EC50 is 2.414-3.196 ng/mL.

● 華美生物ADC靶點(diǎn)蛋白產(chǎn)品列表

產(chǎn)品名稱(chēng) 貨號 靶點(diǎn) 來(lái)源 蛋白標簽
Recombinant Human B-lymphocyte antigen CD19 (CD19), partial (Active) CSB-AP005061HU CD19 Mammalian cell C-terminal Fc-tagged
Recombinant Human B-cell receptor CD22 (CD22), partial (Active) CSB-MP004900HU CD22 Mammalian cell C-terminal 6xHis-tagged
Recombinant Human Programmed cell death 1 ligand 1 (CD274), partial (Active) CSB-MP878942HU1 CD274 Mammalian cell C-terminal hFc-tagged
Recombinant Human CD276 antigen (CD276), partial (Active) CSB-MP733578HU CD276 Mammalian cell C-terminal hFc-Myc-tagged
Recombinant Human Myeloid cell surface antigen CD33 (CD33), partial (Active) CSB-MP004925HU CD33 Mammalian cell C-terminal hFc-Myc-tagged
Recombinant Human Leukocyte surface antigen CD47 (CD47), partial (Active) CSB-AP005201HU CD47 Mammalian cell C-terminal 6xHis-tagged
Recombinant Human Leukocyte surface antigen CD47 (CD47), partial (Active) CSB-AP005211HU CD47 Mammalian cell C-terminal Fc-tagged
Recombinant Human Leukocyte surface antigen CD47 (CD47), partial (Active) CSB-MP004940HU CD47 Mammalian cell C-terminal hFc-tagged
Recombinant Human CD70 antigen (CD70), partial (Active) CSB-MP004954HU1 CD70 Mammalian cell N-terminal 10xHis-tagged
Recombinant Human HLA class II histocompatibility antigen gamma chain (CD74), partial (Active) CSB-MP004956HU1(F2) CD74 Mammalian cell N-terminal 10xHis-tagged
Recombinant Human Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) (E398K) (Active) CSB-MP005165HU CEACAM5 Mammalian cell C-terminal 10xHis-tagged
Recombinant Macaca fascicularis Claudin (CLDN18)-VLPs (Active) CSB-MP4304MOV CLDN18 Mammalian cell N-terminal 6xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Mouse Claudin-18 (Cldn18)-VLPs (Active) CSB-MP005498MO(F3) Cldn18 Mammalian cell N-terminal 6xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Human Claudin-6 (CLDN6)-VLPs (Active) CSB-MP005508HU(A4) CLDN6 Mammalian cell C-terminal 10xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Human Claudin-6 (CLDN6)-VLPs, Fluorescent (Active) CSB-MP005508HU(A4)f4 CLDN6 Mammalian cell C-terminal GFP-tagged (This tag can be tested only under denaturing conditions)
Recombinant Human Delta-like protein 3 (DLL3), partial (Active) CSB-MP882142HU DLL3 Mammalian cell C-terminal 6xHis-tagged
Recombinant Macaca fascicularis Delta-like protein 3 (DLL3), partial (Active) CSB-MP3536MOV DLL3 Mammalian cell C-terminal 6xHis-tagged
Recombinant Human Epidermal growth factor receptor (EGFR), partial (Active) CSB-MP007479HU EGFR Mammalian cell N-terminal 10xHis-tagged and C-terminal Myc-tagged
Recombinant Human Receptor tyrosine-protein kinase erbB-3 (ERBB3), partial (Active) CSB-MP007765HU ERBB3 Mammalian cell C-terminal hFc-tagged
Recombinant Human G-protein coupled receptor family C group 5 member D (GPRC5D)-VLPs (Active) CSB-MP882153HU GPRC5D Mammalian cell C-terminal 10xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Human Heat-stable enterotoxin receptor (GUCY2C), partial (Active) CSB-MP010053HU GUCY2C Mammalian cell N-terminal 10xHis-tagged
Recombinant Human Heat-stable enterotoxin receptor (GUCY2C), partial (Active) CSB-MP010053HUd9 GUCY2C Mammalian cell C-terminal hFc-tagged
Recombinant Human Hepatocyte growth factor receptor (MET), partial (Active) CSB-AP003691HU MET Mammalian cell C-terminal Fc-tagged
Recombinant Human Hepatocyte growth factor receptor (MET), partial (Active) CSB-MP013714HU MET Mammalian cell C-terminal hFc-tagged
Recombinant Human B-lymphocyte antigen CD20 (MS4A1)-VLPs (Active) CSB-MP015007HU MS4A1 Mammalian cell C-terminal 10xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Dog B-lymphocyte antigen CD20 (MS4A1)-VLPs (Active) CSB-MP661636DO MS4A1 Mammalian cell C-terminal 10xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Macaca fascicularis Membrane spanning 4-domains A1 (MS4A1)-VLPs (Active) CSB-MP4516MOV MS4A1 Mammalian cell C-terminal 10xHis-tagged (This tag can be tested only under denaturing conditions)
Recombinant Human Mesothelin (MSLN), partial (Active) CSB-MP015044HUc9 MSLN Mammalian cell N-terminal hFc-tagged
Recombinant Human Nectin-4 (NECTIN4), partial (Active) CSB-AP005571HU NECTIN4 Mammalian cell C-terminal 6xHis-tagged
Recombinant Human Nectin-4 (NECTIN4), partial (Active) CSB-MP822274HU NECTIN4 Mammalian cell C-terminal 10xHis-tagged
Recombinant Human Inactive tyrosine-protein kinase transmembrane receptor ROR1 (ROR1), partial (Active) CSB-MP020067HU1d7 ROR1 Mammalian cell C-terminal 10xHis-tagged
Recombinant Human Zinc transporter ZIP6 (SLC39A6), partial (Active) CSB-BP621669HU1 SLC39A6 Baculovirus C-terminal 6xHis-tagged
Recombinant Human Tumor-associated calcium signal transducer 2 (TACSTD2), partial (Active) CSB-MP023072HU1 TACSTD2 Mammalian cell C-terminal hFc-tagged
Recombinant Human Tumor-associated calcium signal transducer 2 (TACSTD2), partial (Active) CSB-MP023072HU2 TACSTD2 Mammalian cell C-terminal 10xHis-tagged
Recombinant Human Tumor-associated calcium signal transducer 2 (TACSTD2), partial (Active) CSB-MP023072HU1d7 TACSTD2 Mammalian cell C-terminal 10xHis-tagged
Recombinant Human Tumor necrosis factor receptor superfamily member 10B (TNFRSF10B), partial (Active) CSB-AP004921HU TNFRSF10B Mammalian cell C-terminal 6xHis-Fc-tagged
Recombinant Human Tumor necrosis factor receptor superfamily member 10B (TNFRSF10B), partial (Active) CSB-AP004931HU TNFRSF10B Mammalian cell C-terminal 6xHis-tagged
Recombinant Mouse Tumor necrosis factor receptor superfamily member 10B (Tnfrsf10b), partial (Active) CSB-AP005041MO Tnfrsf10b Mammalian cell C-terminal Fc-tagged
Recombinant Human Tumor necrosis factor receptor superfamily member 17 (TNFRSF17), partial (Active) CSB-MP023974HU1 Tnfrsf17 Mammalian cell C-terminal hFc-tagged
Recombinant Human Tumor necrosis factor receptor superfamily member 8 (TNFRSF8), partial (Active) CSB-MP023983HU1 TNFRSF8 Mammalian cell N-terminal 10xHis-tagged and C-terminal Myc-tagged
Recombinant Macaca fascicularis Trophoblast glycoprotein (TPBG), partial (Active) CSB-MP024093MOV TPBG Mammalian cell N-terminal 10xHis-tagged and C-terminal Myc-tagged
Recombinant Human Trophoblast glycoprotein (TPBG), partial (Active) CSB-MP024093HUb0 TPBG Mammalian cell N-terminal 10xHis-tagged


參考文獻:

[1] Abuhelwa Z, Alloghbi A, Nagasaka M. A comprehensive review on antibody-drug conjugates (ADCs) in the treatment landscape of non-small cell lung cancer (NSCLC)[J]. Cancer treatment reviews, 2022, 106.