Novel Silyl Ether-Based Acid-Cleavable Antibody-MMAE Conjugates with Appropriate Stability and Efficacy
Abstract
:1. Introduction
2. Results
2.1. Design and Synthesis of Silyl Ether-Based ADC Payload
2.2. Antibody Conjugation
2.3. The Stability Assays of the Conjugate in Plasma
2.4. Effect of Acidity on ADC Stability
2.5. In Vitro Potency Assay
2.6. Trafficking Assay by Fluorescence Microscopy
2.7. Linker Composition on Inhibition of Microtubule Polymerization
2.8. Cell Cycle Effects
2.9. In Vivo Potency in Xenografted Nude Mice
2.10. Hematology Safety Study
3. Discussion
4. Materials and Methods
4.1. Chemistry
4.2. Synthesis of the Silyl Ether-Based Linker-MMAE Conjugate
4.2.1. Synthesis of 1-(2-(2-hydroxyethoxy)ethyl)-1H-pyrrole-2,5-dione (2)
4.2.2. Synthesis of 4-(((2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethoxy)diisopropylsilyl)-oxy) benzaldehyde (3)
4.2.3. Synthesis of 1-(2-(2-(((4-(hydroxymethyl)phenoxy)diisopropylsilyl)oxy)ethoxy)ethyl)-1H- pyrrole-2,5-dione (4)
4.2.4. Synthesis of 4-(((2-(2-(2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)ethoxy)ethoxy)diisopropylsilyl)-oxy) benzyl(4-nitrophenyl) carbonate (5)
4.2.5. Synthesis of the Linker-MMAE Conjugate (Compound 6)
4.3. Preparation of Reagents and Antibody-Drug Conjugates
4.4. The Stability Assays of Linker and ADC in Plasma
4.5. Effect of Acidity Difference on the Drug Release Process
4.6. Evaluation of ADC for Cancer Cell Killing in Vitro
4.7. Endocytosis and Transport for the Silyl Ether-Based ADC
4.8. Microtubule Polymerization Assay
4.9. Cell Cycle Arrest in HER2-Positive Cancer Cells
4.10. In Vivo Antitumor Activity in Human Gastric Xenograft Tumors
4.11. Hematology Analysis during Treatment
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Cell Lines | HER2 Status | ADC | mil40 | MMAE | |||
---|---|---|---|---|---|---|---|
IC50 (nM) | Max. Inhibition | IC50 (nM) | Max. Inhibition | IC50 (nM) | Max. Inhibition | ||
BT-474 | HER2+++ | 0.170 | 50.50% | 0.323 | 61.25% | 0.400 | 40.65% |
NCI-N87 | HER2+++ | 0.028 | 93.54% | 1.157 | 45.80% | 0.369 | 68.22% |
MDA-MB-453 | HER2++ | 0.101 | 87.30% | 0.315 | 34.80% | 0.295 | 83.79% |
MCF-7 | HER2− | 7.742 | 47.8% | >1000 | - | 0.688 | 77.64% |
MDA-MB-231 | HER2− | >1000 | - | >1000 | - | 1.156 | 53.4% |
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Wang, Y.; Fan, S.; Xiao, D.; Xie, F.; Li, W.; Zhong, W.; Zhou, X. Novel Silyl Ether-Based Acid-Cleavable Antibody-MMAE Conjugates with Appropriate Stability and Efficacy. Cancers 2019, 11, 957. https://doi.org/10.3390/cancers11070957
Wang Y, Fan S, Xiao D, Xie F, Li W, Zhong W, Zhou X. Novel Silyl Ether-Based Acid-Cleavable Antibody-MMAE Conjugates with Appropriate Stability and Efficacy. Cancers. 2019; 11(7):957. https://doi.org/10.3390/cancers11070957
Chicago/Turabian StyleWang, Yanming, Shiyong Fan, Dian Xiao, Fei Xie, Wei Li, Wu Zhong, and Xinbo Zhou. 2019. "Novel Silyl Ether-Based Acid-Cleavable Antibody-MMAE Conjugates with Appropriate Stability and Efficacy" Cancers 11, no. 7: 957. https://doi.org/10.3390/cancers11070957