Biological and mutational analyses of CXCR4-antagonist interactions and design of new antagonistic analogs
Qian Meng1, Ruohan Zhu1, Yujia Mao1
1School of Life Sciences, Tsinghua University, Beijing 100084, China.
Abstract:
The chemokine receptor CXCR4 has become an attractive therapeutic target for HIV-1 infection, hematopoietic stem cell mobilization, and cancer metastasis. A wide variety of synthetic antagonists of CXCR4 have been developed and studied for a growing list of clinical applications. To compare the biological effects of different antagonists on CXCR4 functions and their common and/or distinctive molecular interactions with the receptor, we conducted head-to-head comparative cell-based biological and mutational analyses of the interactions with CXCR4 of eleven reported antagonists, including HC4319, DV3, DV1, DV1 dimer, V1, vMIP-II, CVX15, LY2510924, IT1t, AMD3100, and AMD11070 that were representative of different structural classes of D-peptides, L-peptide, natural chemokine, cyclic peptides, and small molecules. The results were rationalized by molecular modeling of CXCR4-antagonist interactions from which the common as well as different receptor binding sites of these antagonists were derived, revealing a number of important residues such as W94, D97, H113, D171, D262, and E288, mostly of negative charge. To further examine this finding, we designed and synthesized new antagonistic analogs by adding positively charged residues Arg to a D-peptide template to enhance the postulated charge-charge interactions. The newly designed analogs displayed significantly increased binding to CXCR4, which supports the notion that negatively charged residues of CXCR4 can engage in interactions with moieties of positive charge of the antagonistic ligands. The results from these mutational, modeling and new analog design studies shed new insight into the molecular mechanisms of different types of antagonists in recognizing CXCR4 and guide the development of new therapeutic agents.
Insights
Researchers compared eleven CXCR4 antagonists, revealing key receptor interactions. New analogs with positive charges showed enhanced binding, guiding future therapeutic development for HIV, stem cell mobilization, and cancer.
Area of Science:
- Medicinal Chemistry
- Molecular Biology
- Pharmacology
Background:
- CXCR4 receptor antagonists are crucial for HIV-1 infection, stem cell mobilization, and cancer metastasis treatments.
- Numerous synthetic CXCR4 antagonists have been developed, necessitating comparative studies of their efficacy and interaction mechanisms.
Purpose of the Study:
- To compare the biological effects and molecular interactions of eleven diverse CXCR4 antagonists.
- To elucidate common and distinct binding sites and identify key residues involved in CXCR4 antagonism.
Main Methods:
- Head-to-head comparative cell-based biological assays.
- Mutational analyses of CXCR4-antagonist interactions.
- Molecular modeling to rationalize binding sites and interactions.
- Design and synthesis of novel antagonistic analogs.
Main Results:
- Eleven antagonists, representing diverse structural classes, showed varied interactions with CXCR4.
- Molecular modeling identified key receptor residues (e.g., W94, D97, H113, D171, D262, E288), many negatively charged, involved in binding.
- Newly synthesized D-peptide analogs with added positive charges (Arg) exhibited significantly enhanced CXCR4 binding.
Conclusions:
- The study provides new insights into the molecular mechanisms underlying CXCR4 recognition by different antagonist classes.
- Charge-charge interactions between negatively charged CXCR4 residues and positively charged ligand moieties are critical for binding.
- Findings guide the rational design of novel, more effective CXCR4-targeting therapeutic agents.
