Evaluation of an ester-linked immunosuppressive payload: A case study in understanding the stability and cleavability of ester-containing ADC linkers.
Evaluation of an ester-linked immunosuppressive payload: A case study in understanding the stability and cleavability of ester-containing ADC linkers.
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DOI：
10.1016/j.bmcl.2022.128953
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发表时间：
2022-11-01
影响因子：
2.7
通讯作者：
Tumey, L. Nathan
中科院分区：
文献类型：
--
作者：
Jackson, Courtney P.;Fang, Siteng;Benjamin, Samantha R.;Alayi, Tchilabalo;Hathout, Yetrib;Gillen, Sarah M.;Handel, Jillian P.;Brems, Brittany M.;Howe, Justin M.;Tumey, L. Nathan
尽管酯在前药应用中具有价值，但由于人酯酶的普遍存在和混杂性质，通常避免在抗体-药物-缀合物（ADC）有效载荷和接头中使用酯。ADC通常具有长的循环半衰期（3-7天），这使得它们对酯酶介导的代谢敏感。此外，很大程度上不清楚溶酶体和胞质酯酶是否在ADC内化后切割含酯的接头。由于我们对免疫调节剂的靶向递送感兴趣，我们的团队最近制备了一系列酯连接的地塞米松ADC。在此，我们报告了我们对这些ADC的功能活性的研究，特别关注它们在各种生物环境中的催化作用。我们发现，酯是有选择性的，但无效的细胞摄取后裂解，可能是由胞质酯酶。溶酶体催化剂研究表明，尽管有很强的蛋白水解活性，但在溶酶体中很少发生含酯接头的裂解。然而，携带酯连接的有效载荷的ADC在各种免疫抑制测定中是有活性的，表明细胞溶质裂解正在发生。这通过细胞裂解后有效载荷的LCMS定量来证实。最后，在小鼠和人血浆中评价酯键的稳定性。我们发现，类似于其他报告，有一个显着的位点依赖性的切割。连接在高度暴露的位点，如443 C，酯在血浆中迅速裂解，而酯在更受阻的网站，如在334 C，没有。总之，这些结果有助于解开酯掺入ADC接头的复杂性，并为其在ADC应用中的实用性铺平道路。
In spite of their value in prodrug applications, the use of esters in antibody-drug-conjugate (ADC) payloads and linkers has generally been avoided due to the ubiquitous and promiscuous nature of human esterases. ADCs generally have a long circulating half life (3–7 days) that makes them susceptible to esterase-mediated metabolism. Moreover, it is largely unclear whether lysosomal and cytosolic esterases cleave ester-containing linkers upon ADC internalization. Due to our interest in the targeted delivery of immune-modulators, our team has recently prepared a series of ester-linked dexamethasone ADCs. Herein, we report our studies of the functional activity of these ADCs, with a particular focus on their catabolism in various biological milieu. We found that esters are selectively but inefficiently cleaved upon cellular uptake, likely by cytosolic esterases. Lysosomal catabolism studies indicate that, in spite of the strong proteolytic activity, very little cleavage of ester-containing linkers occurs in the lysosome. However, ADCs bearing the ester-linked payloads are active in various immune-suppressive assays, suggesting that cytosolic cleavage is taking place. This was confirmed through LCMS quantitation of the payload following cell lysis. Finally, the stability of the ester linkage was evaluated in mouse and human plasma. We found, similar to other reports, there is a significant site-dependence on the cleavage. Esters attached at highly exposed sites, such as 443C, were rapidly cleaved in plasma while esters at more hindered sites, such at 334C, were not. Together, these results help to unravel the complexities of ester-incorporation into ADC linkers and pave a path forward for their utility in ADC applications.
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影响因子：
12.8
作者：
Ono, K.;Sato, T.;Iyama, S.;Tatekoshi, A.;Hashimoto, A.;Kamihara, Y.;Horiguchi, H.;Kikuchi, S.;Kawano, Y.;Takada, K.;Hayashi, T.;Miyanishi, K.;Sato, Y.;Takimoto, R.;Kobune, M.;Kato, J.
通讯作者：
Kato, J.
影响因子：
4.7
作者：
Dubowchik, GM;Firestone, RA;Trail, PA
通讯作者：
Trail, PA
影响因子：
4.4
作者：
Lux, Anja;Yu, Xiaojie;Nimmerjahn, Falk
通讯作者：
Nimmerjahn, Falk
影响因子：
4.5
作者：
Tumey, L. Nathan;Li, Fengping;Tchistiakova, Lioudmila
通讯作者：
Tchistiakova, Lioudmila
影响因子：
4.9
作者：
Elgersma, Ronald C.;Coumans, Ruud G. E.;Beusker, Patrick H.
通讯作者：
Beusker, Patrick H.