Bifunctional Compound for Targeted Degradation of the Immune Checkpoint Protein PD-L1

doi:10.6023/A24020042

Acta Chimica Sinica ›› 2024, Vol. 82 ›› Issue (6): 613-620.DOI: 10.6023/A24020042 Previous Articles Next Articles

Article

双功能化合物靶向降解免疫检查点蛋白PD-L1

陈从丽^a^,^b, 师怀怀^b, 郝芮^b, 房丽晶^b^,^*(), 徐华栋^a^,^*()

a 常州大学药学院常州 213164
b 中国科学院深圳先进技术研究院生物医药与生物技术研究所深圳 518055

投稿日期:2024-02-01 发布日期:2024-05-04
基金资助:
国家自然科学基金(22177015); 江苏省自然科学基金(BK20211334); 深圳市科委项目(JCYJ20220818101404010); 深圳市科委项目(JCYJ20220818100412028)

Bifunctional Compound for Targeted Degradation of the Immune Checkpoint Protein PD-L1

Congli Chen^a^,^b, Huaihuai Shi^b, Rui Hao^b, Lijing Fang^b,*(), Huadong Xu^a,*()

a School of Pharmacy, Changzhou University, Changzhou 213164
b Institute of Biomedicine and Biotechnology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055

Received:2024-02-01 Published:2024-05-04
Contact: * E-mail: lj.fang@siat.ac.cn; hdxu@cczu.edu.cn
Supported by:
National Natural Science Foundation of China(22177015); Natural Science Foundation of Jiangsu Province(BK20211334); Shenzhen Science and Technology Program(JCYJ20220818101404010); Shenzhen Science and Technology Program(JCYJ20220818100412028)

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Abstract

Programmed death ligand 1 (PD-L1) is a protein overexpressed in numerous tumor cells as an essential immune checkpoint that can facilitate immune escape of tumor cells through binding to PD-1 on T cells. Compared to antibody drugs targeting PD-L1 or PD-1, which are limited in clinical application due to their immunogenicity, high cost and low oral bioavailability, small molecule inhibitors exhibit better tissue and tumor penetration ability, as well as higher bioavailability. However, traditional small molecule inhibitors often face drug resistance issues and are difficult to inhibit protein-protein interactions. Targeted protein degradation (TPD) technologies achieve a more comprehensive suppression of protein activity by degrading the target protein, thereby eliminating its functions entirely. The strategy not only reduces the drug resistance but also enhances the therapeutic efficacy. In the previous study, we have developed the integrin-facilitated lysosome degradation (IFLD) strategy to degrade extracellular and cell membrane proteins by using bifunctional compounds containing a target protein-binding domain, and a cyclic RGD peptide as the integrin-binding domain, connected via a linker. The cyclic RGD peptide, which incorporates the sequence arginine-glycine-aspartic acid (RGD), has been extensively employed for the development of targeted drug delivery systems. It specifically recognizes the integrin α_vβ₃, which is overexpressed in a variety of tumor cells and plays a significant role in tumor-targeted drug delivery. Considering the instability of the RGD peptide, we designed and synthesized a small molecule compound, Gua-Azide, in this study to mimick the RGD (Arg-Gly-Asp) peptide to bind with α_vβ₃ integrin. Subsequently, we constructed a new IFLD degrader targeting PD-L1, BMS-Gua, through the conjugation of Gua-Azide with BMS-8, a small molecule with high binding affinity to PD-L1, by using click chemistry. The western blotting and immunofluorescence analysis verified that BMS-Gua, a bifunctional molecule degrader, could effectively induce the endocytosis and lysosomal degradation of PD-L1 through an integrin-dependent pathway, warranting further investigation for the development of tumor immunotherapy drugs.

Key words: targeted protein degradation, IFLD strategy, bifunctional degrader, PD-L1, cancer immunotherapy

Cite this article

Congli Chen, Huaihuai Shi, Rui Hao, Lijing Fang, Huadong Xu. Bifunctional Compound for Targeted Degradation of the Immune Checkpoint Protein PD-L1[J]. Acta Chimica Sinica, 2024, 82(6): 613-620.

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Fig. & Tab. 6

Figure 1. Comparison of the structures of c(RGDyK) and Gua-Azide

Scheme 1. Synthetic route of Gua-Azide

Figure 2. The stability of Gua-Azide and c(RGDyK) in simulated gastric fluidc

Figure 3. Interactions between Gua-Azide and integrin αVβ3 was assessed by ITC

Scheme 2. Synthetic route of BMS-Gua

Figure 4. BMS-Gua mediated degradation of PD-L1 on the cell membrane. (A) Western blot analysis of PD-L1 protein levels in MDA-MB-231 cells treated with BMS-Gua and BMS-L1-RGD (25 nmol/L, 8 h). (B) Western blot analysis of BMS-Gua-mediated PD-L1 degradation at indicated concentration, or indicated time. (C) Western blot analysis of PD-L1 protein levels in MDA-MB-231 cells treated with 25 nmol/L BMS-Gua for 24 h along with 100 μmol/L Bafilomycin A1 or 5 μmol/L MG132. (D) Effects of competitive inhibition of Gua-Azide-integrin binding on BMS-Gua-mediated PD-L1 degradation, analyzed by Western blot. (E) Immunofluorescence analysis of PD-L1 (red) degradation on cell membrane. HeLa cells stably expressing PD-L1 were treated with BMS-Gua, BMS-8, Gua-Azide, as well as a combination of BMS-8 and Gua-Azide. (F) Effects of competitive inhibition of Gua-Azide-integrin binding on BMS-Gua mediated PD-L1 degradation (red), analyzed by fluorescence imaging. (G) Fluorescence signals indicating the colocalization of PD-L1 (green) with the early endosome marker (Rab5, red) in human cervical carcinoma (HeLa) cells. The nuclei were labeled by DAPI (blue). For (E), (F) and (G), scale bar, 10 μm

References 24

[1]	Zhang, X. Q.; Cheng, C.; Hou, J. Y.; Qi, X. Y.; Wang, X.; Han, P.; Yang, X. M. Cell. Mol. Immunol. 2019, 16, 392.
[2]	Gou, Q.; Dong, C.; Xu, H. H.; Khan, B.; Jin, J. H.; Liu, Q.; Shi, J. J.; Hou, Y. Z. Cell Death Dis. 2020, 11, 955.
[3]	Su, W.; Tan, M. X.; Wang, Z. H.; Zhang, J.; Huang, W. P.; Song, H. H.; Wang, X. Y.; Ran, H. T.; Gao, Y. F.; Nie, G. J.; Wang, H. Angew. Chem., Int. Ed. 2023, 62, e202218128.
[4]	Yang, Z. J.; Huang, C. J.; Wang, M. X. Oncol. Prog. 2020, 18, 772. (in Chinese)
	(杨占菊, 黄长江, 王名雪, 癌症进展, 2020, 18, 772.)
[5]	Sathe, G.; Sapkota, G. P. Trends Pharmacol. Sci. 2023, 44, 786.
[6]	Sun, X. Y.; Gao, H. Y.; Yang, Y. Q.; He, M.; Wu, Y.; Song, Y. G.; Tong, Y.; Rao, Y. Signal Transduct. Tar. 2019, 4, 64.
[7]	He, M.; Cao, C. G.; Ni, Z. H.; Liu, Y. B.; Song, P. L.; Hao, S.; He, Y. N.; Sun, X. Y.; Rao, Y. Signal Transduct. Tar. 2022, 7, 181.
[8]	Zhang, C.; He, Y.; Sun, X.; Wei, W.; Liu, Y.; Rao, Y. Acta Mater. Med. 2023, 2, 409.
[9]	Pei, J. P.; Pan, X. L.; Wang, A. X.; Shuai, W.; Bu, F. Q.; Tang, P.; Zhang, S.; Zhang, Y. W.; Wang, G.; Ouyang, L. Chem. Commun. 2021, 57, 13194.
[10]	Li, Z. Y.; Zhu, C. G.; Ding, Y.; Fei, Y. Y.; Lu, B. X. Autophagy 2020, 16, 185.
[11]	Ji, C. H.; Kim, H. Y.; Lee, M. J.; Heo, A. J.; Park, D. Y.; Lim, S.; Shin, S.; Yang, W. S.; Jung, C. A.; Kim, K. Y.; Jeong, E. H.; Park, S. H.; Kim, S. B.; Lee, S. J.; Na, J. E.; Kang, J. I.; Chi, H. M.; Kim, H. T.; Kim, Y. K.; Kim, B. Y.; Kwon, Y. T. Nat. Commun. 2022, 13, 904.
[12]	Banik, S. M.; Pedram, K.; Wisnovsky, S.; Ahn, G.; Riley, N. M.; Bertozzi, C. R. Nature 2020, 584, 291.
[13]	Cotton, A. D.; Nguyen, D. P.; Gramespacher, J. A.; Seiple, I. B.; Wells, J. A. J. Am. Chem. Soc. 2021, 143, 593. doi: 10.1021/jacs.0c10008 pmid: 33395526
[14]	Zhang, H.; Han, Y.; Yang, Y. F.; Lin, F.; Li, K. X.; Kong, L. H.; Liu, H. X.; Dang, Y. J.; Lin, J.; Chen, P. R. J. Am. Chem. Soc. 2021, 143, 16377. doi: 10.1021/jacs.1c08521 pmid: 34596400
[15]	Wu, Y. Q.; Lin, B. Q.; Lu, Y. Z.; Li, L.; Deng, K. Y.; Zhang, S. H.; Zhang, H. M.; Yang, C. Y.; Zhu, Z. Angew. Chem., Int. Ed. 2023, 62, e202218106.
[16]	Zheng, J. W.; He, W. Y.; Li, J.; Feng, X. J.; Li, Y. Y.; Cheng, B. H.; Zhou, Y. M.; Li, M. Q.; Liu, K.; Shao, X. M.; Zhang, J. C.; Li, H. C.; Chen, L.; Fang, L. J. J. Am. Chem. Soc. 2022, 144, 21831.
[17]	Nieberler, M.; Reuning, U.; Reichart, F.; Notni, J.; Wester, H. J.; Schwaiger, M.; Weinmüller, M.; Räder, A.; Steiger, K.; Kessler, H. Cancers 2017, 9, 116.
[18]	Hatley, R. J. D.; Macdonald, S. J. F.; Slack, R. J.; Le, J.; Ludbrook, S. B.; Lukey, P. T. Angew. Chem., Int. Ed. 2018, 57, 3298.
[19]	Danhier, F.; Le Breton, A.; Préat, V. Mol. Pharmaceut. 2012, 9, 2961.
[20]	Chen, T. K.; Li, Q.; Liu, Z. L.; Chen, Y.; Feng, F.; Sun, H. P. Eur. J. Med. Chem. 2019, 161, 378.
[21]	Sulyok, G. A. G.; Gibson, C.; Goodman, S. L.; Hölzemann, G.; Wiesner, M.; Kessler, H. J. Med. Chem. 2001, 44, 1938. pmid: 11384239
[22]	Chai, Z. L.; Ran, D. N.; Lu, L. W.; Zhan, C. Y.; Ruan, H. T.; Hu, X. F.; Xie, C.; Jiang, K.; Li, J. Y.; Zhou, J. F.; Wang, J.; Zhang, Y. Y.; Fang, R. H.; Zhang, L. F.; Lu, W. Y. ACS Nano 2019, 13, 5591.
[23]	Wang, X.; Zhang, X. R.; Huang, Z. Y.; Fan, X. Y.; Chen, P. Acta Chim. Sinica 2021, 79, 406. (in Chinese) doi: 10.6023/A20110530
	(汪欣, 张贤睿, 黄宗煜, 樊新元, 陈鹏, 化学学报, 2021, 79, 406.) doi: 10.6023/A20110530
[24]	Guzik, K.; Zak, K. M.; Grudnik, P.; Magiera, K.; Musielak, B.; Törner, R.; Skalniak, L.; Dömling, A.; Dubin, G.; Holak, T. A. J. Med. Chem. 2017, 60, 5857.

双功能化合物靶向降解免疫检查点蛋白PD-L1

Bifunctional Compound for Targeted Degradation of the Immune Checkpoint Protein PD-L1

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