Meneghello Raphael, Rustiguel Joane K, de Araújo Evandro Ares, de Felício Rafael, Fernandes Arthur Zanetti N, Ferreira Everton L F, Gubiani Juliana R, Takeda Agnes A S, Araujo Amanda, de Lima Silva Caio C, Bertonha Ariane F, Urano Raquel P M, Trindade Daniel M, Cunha Thiago M, Cardoso Alisson C, Berlinck Roberto G S, Nascimento Andrey F Ziem, Trivella Daniela B B
Brazilian Biosciences National Laboratory (LNBio), Brazilian Centre for Research in Energy and Materials (CNPEM), Campinas, SP 13083-970, Brazil.
Brazilian Synchrotron Light Source National Laboratory (LNLS), Brazilian Centre for Research in Energy and Materials (CNPEM), Campinas, SP 13083-970, Brazil.
Acta Crystallogr F Struct Biol Commun. 2025 May 1;81(Pt 5):179-192. doi: 10.1107/S2053230X25001542. Epub 2025 Apr 16.
Nature is a rich and largely untapped reservoir of small molecules, the latter historically being the main source of new drugs. Three-dimensional structures of proteins in complex with small-molecule ligands represent key information to progress drug-discovery projects, in particular in the hit-to-lead phase. High-throughput crystallography has been of extensive use in recent years, especially to obtain crystallographic complexes of synthetic ligands and fragments. However, the process of discovering novel bioactive natural products has experienced limitations that have long prevented large drug-discovery programs using this outstanding source of molecules. Recent technologies have contributed to the re-emergence of natural products in modern drug discovery. We present the use of high-throughput protein crystallography to directly capture bioactive natural products from unpurified biota chemical samples using protein crystals. These routines, which are currently in use at the Brazilian Centre for Research in Energy and Materials (CNPEM), are introduced with a description of crystal preparation, automated data collection and processing at the MANACÁ beamline (Sirius, LNLS, CNPEM), along with case examples of bioactive natural product capture using protein crystals. The usefulness of this pipeline, which accelerates the discovery and structural elucidation of both known and previously unknown bioactive natural products, paves the way for the development of innovative therapeutic agents, thus contributing to the new era of natural product-based drug discovery.
自然界是小分子的丰富宝库，且在很大程度上尚未被开发利用，而小分子一直以来都是新药的主要来源。蛋白质与小分子配体复合物的三维结构是推进药物研发项目的关键信息，尤其是在从活性分子到先导化合物的阶段。近年来，高通量晶体学得到了广泛应用，特别是用于获取合成配体和片段的晶体学复合物。然而，发现新型生物活性天然产物的过程存在一些局限性，长期以来阻碍了利用这一出色分子来源开展大型药物研发项目。近期的技术推动了天然产物在现代药物研发中的再度兴起。我们展示了利用高通量蛋白质晶体学，通过蛋白质晶体直接从未纯化的生物群化学样品中捕获生物活性天然产物。本文介绍了目前巴西能源和材料研究中心（CNPEM）所使用的这些方法，包括在MANACÁ光束线（天狼星，巴西国家同步辐射实验室，CNPEM）进行晶体制备、自动数据收集和处理的过程，以及使用蛋白质晶体捕获生物活性天然产物的实例。这条流程加快了已知和此前未知的生物活性天然产物的发现及结构解析，为创新治疗药物的开发铺平了道路，从而为基于天然产物的药物研发新时代做出了贡献。
