Zhao, Yanbo;
(2023)
Expanding the scope of Next Generation Maleimides for Antibody Conjugation.
Doctoral thesis (Ph.D), UCL (University College London).
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Abstract
Antibody drug conjugates (ADCs) are increasingly promising targeted therapies for cancer treatments, due to the combination of antibodies with tumour selectivity and cytotoxic drugs. Current strategies to construct ADCs suffer from heterogeneity, complexity, and high costs. Dibromomaleimides (DBMs), a class of next generation maleimides (NGMs), have shown ability to site-selectively bridge antibody disulfide bonds, delivering robustly stable conjugates following maleimide hydrolysis. This work expands DBM scope by developing trifunctional DBMs built around a lysine core, introducing multiple functionalities (such as fluorophores) onto an antibody, which would be of significant interest for treatments of complex diseases. However, a problem associated with DBM-based disulfide bridging arises as disulfide scrambling where incorrectly bridged disulfides observed in antibody hinge region, thus forming two antibody isomers with limited homogeneity. To resolve this issue, in situ NGMbased bioconjugation has been conducted to enable simultaneous disulfide reduction and bridging. A variety of dithiomaleimides (DTMs) has been explored to attempt to minimise cross-reactivity to TCEP, a typical disulfide reducing agent. Compared to highly reactive DBMs, these DTMs have attenuated reactivity, and thus can be applied during the reduction. Upon disulfide reduction, the presence of these DTMs enables an efficient bridging and leads to a reduction in scrambled disulfide bonds compared to conventional DBM-based bioconjugation. However, the resultant conjugates also involve mis-bridged antibody species, therefore, NGM-based in situ bioconjugation still needs optimisation. Lastly, investigations on benzeneselenols as novel antibody reducing agents have been conducted. Traditional benzeneselenols show good reducing ability but suffer from the poor aqueous solubility and malodour. A new generation of benzeneselenols with improved bioconjugation properties have been developed, through the synthesis of substituted aryl derivatives with improved solubility for better compatibility with biological conditions. Overall, this work expands the scope of DBMs and explores methods to generate more homogeneous antibody conjugates using DBMs and related reagents.
| Type: | Thesis (Doctoral) |
|---|---|
| Qualification: | Ph.D |
| Title: | Expanding the scope of Next Generation Maleimides for Antibody Conjugation |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| Additional information: | © The Author(s). This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/. |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Chemistry |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10173516 |
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