mRNA display-enabled discovery of proximity-triggered covalent peptide-drug conjugates.

Ruixuan WANG; Siqi RAN; Jiabei GUO; Da HU; Xiang FENG; Jixia ZHOU; Zhanzhi ZHANG; Futian LIANG; Jiamin SHANG; Lingxin BU; Kaiyi WANG; Junyi MAO; Huixin LUO; Rui WANG

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mRNA display-enabled discovery of proximity-triggered covalent peptide-drug conjugates.

Author: Ruixuan WANG ¹ ; Siqi RAN ¹ ; Jiabei GUO ¹ ; Da HU ¹ ; Xiang FENG ¹ ; Jixia ZHOU ¹ ; Zhanzhi ZHANG ¹ ; Futian LIANG ¹ ; Jiamin SHANG ¹ ; Lingxin BU ¹ ; Kaiyi WANG ¹ ; Junyi MAO ¹ ; Huixin LUO ¹ ; Rui WANG ¹
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1. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Publication Type:Journal Article
Keywords: Covalent peptide; Flexizyme; Fluorosulfate-l-tyrosine; Macrocyclic peptide; Nectin-4; Peptide–drug conjugate; Sulfur(VI) fluoride exchange; mRNA display
From: Acta Pharmaceutica Sinica B 2025;15(10):5474-5485
CountryChina
Language:English
Abstract: Peptide-drug conjugates (PDCs) have emerged as a promising modality in precision oncology, enabling targeted delivery of cytotoxic payloads while minimizing off-target toxicity. The integration of covalent warheads, such as those based on sulfur(VI) fluoride exchange (SuFEx) chemistry, enhances drug-target residence time and tumor accumulation. However, existing screening methods for covalent peptide (CP) libraries require post-translational warhead conjugation, limiting throughput. Here, we present an integrated mRNA display platform that incorporates covalent warheads during ribosomal synthesis, enabling efficient screening of ultra-diverse covalent macrocyclic peptide libraries (>10¹³ variants). This approach, using site-specific incorporation of N-chloroacetyl-d-phenylalanine and fluorosulfate-l-tyrosine, accelerated the discovery of irreversibly binding (K _i = 3.58 μmol/L) Nectin-4-targeting peptide CP-N1-N₃ via proximity-triggered SuFEx. The peptide was further conjugated to cytotoxic payloads, yielding the covalent PDC CP-N1-MMAE with potent cytotoxicity (IC₅₀ ≈ 43 nmol/L) against MDA-MB-468 cells. This platform establishes a new paradigm for precision covalent drug discovery.