Sortase Enzyme-Mediated Generation of Site-Specifically Conjugated Antibody Drug Conjugates with High In Vitro and In Vivo Potency.
Sortase Enzyme-Mediated Generation of Site-Specifically Conjugated Antibody Drug Conjugates with High In Vitro and In Vivo Potency.
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DOI：
10.1371/journal.pone.0131177
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Grawunder U
中科院分区：
文献类型：
--
作者：
Beerli RR;Hell T;Merkel AS;Grawunder U
近年来，抗体药物偶联物（adc）已被证明是一种高效的抗肿瘤药物，其疗效通常超过传统的单克隆抗体（mab）。adc目前是通过赖氨酸或半胱氨酸侧链将小分子毒素与单抗化学偶联而产生的。这导致adc的异质混合物，其中可变数量的药物与单个抗体结合，并且无法确定结合的位置。因此，目前对进一步开发药物偶联技术有很大的兴趣，特别关注位点特异性有效载荷偶联。在这里，我们提出了一个基于金黄色葡萄球菌分类酶a介导的转肽化反应的酶偶联平台，允许有效地产生adc，毒素以预定义的药物-抗体比例偶联到预定义的位点。为此，引入了两种修饰：首先，通过添加排序酶A识别基序LPETG，在免疫球蛋白重链（IgH）和轻链（IgL）的c端进行修饰；其次，通过添加五甘氨酸肽，对小分子微管蛋白聚合抑制剂单甲基lauristatin E （MMAE）和美丹氨酸进行修饰，从而使它们成为排序酶A介导的转肽化的合适底物。我们证明了抗cd30 ADC Ac10-vcPAB-MMAE的有效生成和表征，Ac10-vcPAB-MMAE是brentuximab vedotin （Adcetris）的酶偶联对应物，以及几种抗her -2 ADC，包括曲妥珠单抗maytansine，曲妥珠单抗emtansine （Kadcyla）的对应物。以这种方式产生的adc显示出与经典偶联物没有区别的体外细胞杀伤活性。此外，当在her -2过表达的卵巢癌异种移植小鼠模型中进行体内测试时，发现酶促生成的曲妥珠单抗-美坦素可导致已建立的肿瘤完全消退，类似于Kadcyla。
Antibody drug conjugates (ADCs) have recently been proven to be highly potent anti-tumor drugs, typically exceeding the efficacy of conventional monoclonal antibodies (mAbs). ADCs are currently produced by chemical conjugation of a small-molecule toxin to the mAb through lysine or cysteine side chains. This leads to heterogeneous mixtures of ADCs in which variable numbers of drugs are conjugated to individual antibodies and in which the site of conjugation cannot be defined. Consequently, there is currently significant interest in further development of drug conjugation technologies, with a particular focus on site-specific payload conjugation. Here, we present an enzymatic conjugation platform based on the S. aureus sortase A-mediated transpeptidation reaction, allowing the efficient generation of ADCs with toxins conjugated to pre-defined sites at pre-defined drug-to-antibody ratios. For this, two modifications were introduced: first, immunoglobulin heavy (IgH) and light (IgL) chains were modified at their C-termini by addition of the sortase A recognition motif LPETG, and second, the small molecule tubulin polymerization inhibitors monomethylauristatin E (MMAE) and maytansine were modified by addition of a pentaglycine peptide, thus making them suitable substrates for sortase A-mediated transpeptidation. We demonstrate efficient generation and characterization of the anti-CD30 ADC Ac10-vcPAB-MMAE, an enzymatically conjugated counterpart of brentuximab vedotin (Adcetris), as well as several anti-HER-2 ADCs including trastuzumab-maytansine, the counterpart of trastuzumab emtansine (Kadcyla). ADCs generated in this manner were found to display in vitro cell killing activities indistinguishable from the classic conjugates. Further, when tested in vivo in a HER-2-overexpressing ovarian cancer xenograft mouse model, enzymatically generated trastuzumab-maytansine was found to lead to complete regression of established tumors, similar to Kadcyla.
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DOI：
10.1038/nrc3239
发表时间：
2012-03-22
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
Pardoll DM
通讯作者：
Pardoll DM
影响因子：
11.2
作者：
Erickson, HK;Park, PU;Blättler, WA
通讯作者：
Blättler, WA
影响因子：
4.7
作者：
Doronina, SO;Mendelsohn, BA;Senter, PD
通讯作者：
Senter, PD
影响因子：
5.8
作者：
Alewine, Christine;Hassan, Raffit;Pastan, Ira
通讯作者：
Pastan, Ira
影响因子：
46.9
作者：
Lyon, Robert P.;Setter, Jocelyn R.;Senter, Peter D.
通讯作者：
Senter, Peter D.