Site-selective antibody modification by cysteine-to-lysine transfer (CLT)

通过半胱氨酸到赖氨酸转移 (CLT) 进行位点选择性抗体修饰

• 批准号: EP/R034621/1
• 负责人: James Baker
• 金额: $ 52.47万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FR034621%2F1
• 关键词: Site; selective; antibody; modification; cysteine

With CRUK recently highlighting that 1 in 2 of us will have cancer in our lifetimes, the requirement for progress in developing new medicines and improved diagnostics for oncology is a high priority for our society. The current state of the art in chemotherapeutics still relies heavily on untargeted cytotoxins, leading to severe side-effects which are intolerable in many cases. In contrast, the attachment of cytotoxins to a delivery agent, which targets the 'warhead' specifically to the tumor, offers the enticing possibility of 'magic-bullet' chemotherapies. Antibody-Drug Conjugates (ADCs) represent amongst the most promising class of such drugs in development, with 3 ADCs achieving clinical approval in the last few years. However, it is widely considered that the molecular construction of ADCs currently is still far from optimal, and that new technologies in this area are urgently required to help enable them to achieve their clinical potential. Most notably current approaches employed to attach the cytotoxic drugs to the antibodies lead to a highly complex mixture of products. The result is a drug which contains a vast number of distinct species, each with a different pharmacological profile. In this project we are proposing to pioneer a new chemical approach for the generation of ADCs, which attaches the drugs at specific locations on the antibody, generating superior homogeneous conjugates. Our strategy, crucially, will not require genetic engineering of the antibodies to incorporate reactive handles, and is thus applicable directly to native 'off-the-shelf' antibodies. This will maximise the accessibility of homogenous ADCs to researchers across the world and ensure that the production yields are maintained as high as possible, ultimately reducing the cost of these relatively complex biopharmaceuticals. We will achieve goal this by targeting specific lysine amino-acids on the surface of the antibodies. This is a challenging aim as there are numerous lysines present; and as such we are proposing to develop new methodology, in which the conjugation reagent is guided to a specific location by a neighbouring cysteine amino-acid. This unprecedented cysteine-to-lysine transfer (CLT) approach will generate amide linkages between the antibody and the drug which are already extremely well characterised and known to be robustly stable in vivo. This will afford a high-level of confidence which will facilitate rapid uptake in the field. Overall this CLT platform will represent an optimum approach for producing ADCs and facilitate the wider success of these exciting targeted therapies.

CRUK 最近强调，我们中有二分之一的人在一生中会患上癌症，因此在开发新药和改进肿瘤学诊断方面取得进展的要求是我们社会的首要任务。目前的化学治疗技术仍然严重依赖非靶向细胞毒素，导致许多情况下无法忍受的严重副作用。相比之下，将细胞毒素附着在递送剂上，将“弹头”特异性地瞄准肿瘤，为“神奇子弹”化疗提供了诱人的可能性。抗体药物偶联物 (ADC) 是开发中最有前途的此类药物之一，过去几年有 3 种 ADC 获得了临床批准。然而，人们普遍认为，目前ADC的分子结构还远未达到最佳状态，迫切需要该领域的新技术来帮助其发挥临床潜力。最值得注意的是，当前用于将细胞毒性药物附着到抗体上的方法导致了高度复杂的产品混合物。其结果是一种药物含有大量不同的物种，每种物种都有不同的药理学特征。 在这个项目中，我们建议开创一种用于生成 ADC 的新化学方法，该方法将药物附着在抗体上的特定位置，生成优质的均质缀合物。至关重要的是，我们的策略不需要对抗体进行基因工程来整合反应性手柄，因此可直接适用于天然的“现成”抗体。这将最大限度地提高世界各地研究人员获得同质 ADC 的机会，并确保产量保持尽可能高，最终降低这些相对复杂的生物制药的成本。 我们将通过靶向抗体表面的特定赖氨酸氨基酸来实现这一目标。这是一个具有挑战性的目标，因为存在大量赖氨酸；因此，我们建议开发新的方法，其中缀合试剂由邻近的半胱氨酸氨基酸引导至特定位置。这种前所未有的半胱氨酸到赖氨酸转移（CLT）方法将在抗体和药物之间产生酰胺键，这些键已经被非常充分地表征并且已知在体内非常稳定。这将提供高度的信心，从而促进该领域的快速采用。 总体而言，该 CLT 平台将代表生产 ADC 的最佳方法，并促进这些令人兴奋的靶向疗法取得更广泛的成功。

项目成果

New Bifunctional Chelators Incorporating Dibromomaleimide Groups for Radiolabeling of Antibodies with Positron Emission Tomography Imaging Radioisotopes.

• DOI: 10.1021/acs.bioconjchem.0c00710
• 发表时间: 2021-07-21
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Farleigh M;Pham TT;Yu Z;Kim J;Sunassee K;Firth G;Forte N;Chudasama V;Baker JR;Long NJ;Rivas C;Ma MT
• 通讯作者: Ma MT

Application of Next-Generation Maleimides (NGMs) to Site-Selective Antibody Conjugation.

下一代马来酰亚胺 (NGM) 在位点选择性抗体缀合中的应用。

• DOI: 10.1007/978-1-4939-9654-4_2
• 发表时间: 2019
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Morais M

Cysteine-to-lysine transfer antibody fragment conjugation

• DOI: 10.1039/c9sc03825f
• 发表时间: 2019-12-21
• 期刊: CHEMICAL SCIENCE
• 影响因子: 8.4
• 作者: Forte, Nafsika;Benni, Irene;Baker, James R.
• 通讯作者: Baker, James R.

Site-selective lysine conjugation methods and applications towards antibody-drug conjugates.

• DOI: 10.1039/d1cc03976h
• 发表时间: 2021-10-14
• 期刊: Chemical communications (Cambridge, England)
• 作者: Haque M;Forte N;Baker JR
• 通讯作者: Baker JR

A Plug-and-Play Platform for the Formation of Trifunctional Cysteine Bioconjugates that also Offers Control over Thiol Cleavability.

• DOI: 10.1021/acs.bioconjchem.1c00057
• 发表时间: 2021-04-21
• 期刊: Bioconjugate chemistry
• 影响因子: 4.7
• 作者: Bahou C;Szijj PA;Spears RJ;Wall A;Javaid F;Sattikar A;Love EA;Baker JR;Chudasama V
• 通讯作者: Chudasama V