SLAS Discovery最新文献_第2页

High-throughput combination screening of Pidnarulex and other G-quadruplex ligands in multi-cell type tumor spheroids Pidnarulex等g -四联体配体在多细胞型肿瘤球体中的高通量联合筛选。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-21 DOI: 10.1016/j.slasd.2025.100284

Thomas S. Dexheimer , Nathan P. Coussens , Thomas Silvers , Poorva Juneja , Eric Jones , Steven D. Gore , Mark W. Kunkel , James H. Doroshow , Beverly A. Teicher

G-quadruplexes (G4s) are four-stranded nucleic acid structures that regulate key cellular processes and represent promising therapeutic targets in oncology. To investigate the therapeutic potential of three G4 ligands—pidnarulex, APTO-253, and BRACO-19—a high-throughput drug combination screen was conducted in thirty-one multi-cell type tumor spheroids derived from patient tumors and established cancer cell lines. These 3D spheroids mimic key features of the tumor microenvironment, comprising malignant, endothelial, and mesenchymal cell populations. Compounds selected for combination screening included agents with mechanistic relevance to G4 biology, such as inhibitors of DNA damage response (DDR), replication stress, and chromatin regulation, based on the proposed roles of G4s in replication and genome stability. Combination responses were assessed using cell viability assays and supported by longitudinal brightfield imaging to monitor spheroid morphology and growth dynamics. Drug interactions were quantified using Bliss independence scores and the volume under the viability surface, providing complementary metrics of synergy and overall response. Among the G4 ligands, pidnarulex demonstrated the broadest single-agent activity, while APTO-253 and BRACO-19 showed limited effects. Model-specific synergy was observed from combinations with inhibitors of PARP, DDR kinases (ATM, ATR, DNA-PK), and cell cycle regulators (WEE1, PIM1). Interestingly, pidnarulex exhibited consistent synergy in one of eight pancreatic adenocarcinoma models (966289-007-R4-J1) across multiple DDR-targeted combinations. Combination interactions were also observed with HDAC inhibitors in a subset of models. Brightfield imaging corroborated enhanced spheroid growth suppression from synergistic combinations. These findings underscore the context-dependent activity of G4 ligands and support the use of integrated functional and imaging-based approaches to characterize potential therapeutic combinations in physiologically relevant 3D cancer models.

g -四链（G4s）是一种调节关键细胞过程的四链核酸结构，是肿瘤学中有前景的治疗靶点。为了研究3种G4配体pidnarulex、APTO-253和braco -19的治疗潜力，我们对31个来自患者肿瘤和已建立的癌细胞系的多细胞型肿瘤球体进行了高通量联合筛选。这些三维球体模拟肿瘤微环境的关键特征，包括恶性、内皮细胞和间充质细胞群。根据G4s在复制和基因组稳定性中的作用，选择的化合物包括与G4生物学机制相关的药物，如DNA损伤反应（DDR）抑制剂、复制应激和染色质调节。通过细胞活力测定和纵向明场成像来监测球体形态和生长动态来评估组合反应。使用Bliss独立性评分和活力表面下的体积来量化药物相互作用，提供协同作用和总体反应的补充指标。在G4配体中，pidnarulex具有最广泛的单药活性，而APTO-253和BRACO-19作用有限。通过与PARP、DDR激酶抑制剂（ATM、ATR、DNA-PK）和细胞周期调节剂（WEE1、PIM1）的联合，观察到模型特异性协同作用。有趣的是，pidnarulex在8种胰腺腺癌模型之一（966289-007-R4-J1）中跨多种ddr靶向组合表现出一致的协同作用。在一些模型中也观察到与HDAC抑制剂的联合相互作用。亮场成像证实了协同组合增强的球体生长抑制。这些发现强调了G4配体的环境依赖性活性，并支持在生理相关的3D癌症模型中使用综合功能和基于成像的方法来表征潜在的治疗组合。

{"title":"High-throughput combination screening of Pidnarulex and other G-quadruplex ligands in multi-cell type tumor spheroids","authors":"Thomas S. Dexheimer , Nathan P. Coussens , Thomas Silvers , Poorva Juneja , Eric Jones , Steven D. Gore , Mark W. Kunkel , James H. Doroshow , Beverly A. Teicher","doi":"10.1016/j.slasd.2025.100284","DOIUrl":"10.1016/j.slasd.2025.100284","url":null,"abstract":"<div><div>G-quadruplexes (G4s) are four-stranded nucleic acid structures that regulate key cellular processes and represent promising therapeutic targets in oncology. To investigate the therapeutic potential of three G4 ligands—pidnarulex, APTO-253, and BRACO-19—a high-throughput drug combination screen was conducted in thirty-one multi-cell type tumor spheroids derived from patient tumors and established cancer cell lines. These 3D spheroids mimic key features of the tumor microenvironment, comprising malignant, endothelial, and mesenchymal cell populations. Compounds selected for combination screening included agents with mechanistic relevance to G4 biology, such as inhibitors of DNA damage response (DDR), replication stress, and chromatin regulation, based on the proposed roles of G4s in replication and genome stability. Combination responses were assessed using cell viability assays and supported by longitudinal brightfield imaging to monitor spheroid morphology and growth dynamics. Drug interactions were quantified using Bliss independence scores and the volume under the viability surface, providing complementary metrics of synergy and overall response. Among the G4 ligands, pidnarulex demonstrated the broadest single-agent activity, while APTO-253 and BRACO-19 showed limited effects. Model-specific synergy was observed from combinations with inhibitors of PARP, DDR kinases (ATM, ATR, DNA-PK), and cell cycle regulators (WEE1, PIM1). Interestingly, pidnarulex exhibited consistent synergy in one of eight pancreatic adenocarcinoma models (966289-007-R4-J1) across multiple DDR-targeted combinations. Combination interactions were also observed with HDAC inhibitors in a subset of models. Brightfield imaging corroborated enhanced spheroid growth suppression from synergistic combinations. These findings underscore the context-dependent activity of G4 ligands and support the use of integrated functional and imaging-based approaches to characterize potential therapeutic combinations in physiologically relevant 3D cancer models.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"37 ","pages":"Article 100284"},"PeriodicalIF":2.7,"publicationDate":"2025-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145357056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative analysis of antibody-mediated loss-of-function versus gene knock-out and knock-down 抗体介导的功能丧失与基因敲除和敲除的比较分析。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-13 DOI: 10.1016/j.slasd.2025.100283

Marie Buck-Wiese , Sally Liechocki , Holger Erfle , Vytaute Starkuviene

In this study we compare three methods for manipulating cell function: RNA interference (RNAi), CRISPR-Cas9 gene knock-out, and antibody-mediated loss-of-function. We have focused on analyzing changes in cell-matrix adhesion via targeting two key regulators, Talin1 (TLN1) and Kindlin-2 (KD2). Adhesion-relevant phenotypic assays revealed distinct temporal onset dynamics for each method. RNAi and CRISPR-Cas9 effectively reduced target mRNA and protein levels. In contrast, antibody transfection induced phenotypic changes without altering target expression, suggesting direct intracellular antibody-target interaction. Transcriptome analysis demonstrated that antibody transfection and CRISPR-Cas9 induced fewer deregulated mRNAs than RNAi. Furthermore, transfected antibodies and sgRNAs shared 30 % and 70 % of deregulated transcripts to their negative controls, respectively. Whereas only 10 % of overlap was recorded between targeting and control siRNAs. Our findings emphasize the importance of considering method-specific temporal dynamics of on-target phenotype appearance and off-target manifestation. Additionally, they highlight intracellular delivered antibodies as a valuable alternative for validating and complementing genetic approaches.

在这项研究中，我们比较了三种操纵细胞功能的方法：RNA干扰（RNAi）、CRISPR-Cas9基因敲除和抗体介导的功能丧失。我们专注于通过靶向两个关键调节因子Talin1 （TLN1）和Kindlin-2 （KD2）来分析细胞-基质粘附的变化。黏附相关的表型分析揭示了每种方法不同的时间发病动态。RNAi和CRISPR-Cas9有效降低了靶mRNA和蛋白水平。相反，抗体转染诱导表型改变而不改变靶标表达，提示细胞内抗体-靶标直接相互作用。转录组分析表明，与RNAi相比，抗体转染和CRISPR-Cas9诱导的失调mrna较少。此外，转染的抗体和sgrna分别与阴性对照共享30%和70%的解除调控转录本。而在靶向sirna和控制sirna之间只有10%的重叠。我们的研究结果强调了考虑靶上表型和脱靶表现的方法特异性时间动态的重要性。此外，他们强调细胞内递送抗体作为验证和补充遗传方法的有价值的替代方法。

{"title":"Comparative analysis of antibody-mediated loss-of-function versus gene knock-out and knock-down","authors":"Marie Buck-Wiese , Sally Liechocki , Holger Erfle , Vytaute Starkuviene","doi":"10.1016/j.slasd.2025.100283","DOIUrl":"10.1016/j.slasd.2025.100283","url":null,"abstract":"<div><div>In this study we compare three methods for manipulating cell function: RNA interference (RNAi), CRISPR-Cas9 gene knock-out, and antibody-mediated loss-of-function. We have focused on analyzing changes in cell-matrix adhesion via targeting two key regulators, Talin1 (TLN1) and Kindlin-2 (KD2). Adhesion-relevant phenotypic assays revealed distinct temporal onset dynamics for each method. RNAi and CRISPR-Cas9 effectively reduced target mRNA and protein levels. In contrast, antibody transfection induced phenotypic changes without altering target expression, suggesting direct intracellular antibody-target interaction. Transcriptome analysis demonstrated that antibody transfection and CRISPR-Cas9 induced fewer deregulated mRNAs than RNAi. Furthermore, transfected antibodies and sgRNAs shared 30 % and 70 % of deregulated transcripts to their negative controls, respectively. Whereas only 10 % of overlap was recorded between targeting and control siRNAs. Our findings emphasize the importance of considering method-specific temporal dynamics of on-target phenotype appearance and off-target manifestation. Additionally, they highlight intracellular delivered antibodies as a valuable alternative for validating and complementing genetic approaches.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"37 ","pages":"Article 100283"},"PeriodicalIF":2.7,"publicationDate":"2025-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145304879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

From Large to Small Cytokine Receptor Antagonists 从大到小细胞因子受体拮抗剂。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-04 DOI: 10.1016/j.slasd.2025.100280

Pawel Leznicki, Karl Deacon, Bohdan Waszkowycz, John F Unitt

Here, we review general trends in biological and small molecule cytokine drug discovery, highlighting key learnings from two successful case studies (TNFα and IL-17), which have helped shape our understanding of how best to identify new small molecule cytokine antagonists.

Over the past three decades, biological drugs have revolutionized the treatment of a wide range of diseases, from oncology to autoimmune diseases. Their ability to modulate extracellular targets, often inaccessible to conventional small molecule drugs due to their complex protein-protein interactions, has enabled the clinical validation of numerous novel cytokine targets.

Despite their success, biological drugs have significant limitations. Challenges include poor oral bioavailability, tissue penetration and access to intracellular drug targets, as well as high manufacturing costs. These constraints have catalyzed efforts to develop small molecule equivalents that replicate the therapeutic efficacy of biologicals while overcoming their poor delivery and high production costs.

Over the past twenty years, advances in structural biology, computational modeling, disease biology, hit-finding technologies, and medicinal chemistry have converged to collectively enable the identification and subsequent progression of small molecule cytokine modulators into clinical development.

Looking ahead, we speculate on the future drug discovery landscape of this field with the likely emergence of small molecules drugs for a range of clinically validated cytokines like TSLP and TL1A. This evolution will be accelerated by the advent of novel modalities like extracellular degraders, oral peptide drugs and the development of next-generation biological drugs with multi-valency and improved delivery.

在这里，我们回顾了生物和小分子细胞因子药物发现的一般趋势，强调了两个成功案例研究（TNFα和IL-17）的关键学习，这有助于我们理解如何最好地识别新的小分子细胞因子拮抗剂。在过去的三十年里，生物药物已经彻底改变了从肿瘤到自身免疫性疾病等一系列疾病的治疗。它们调节细胞外靶点的能力，由于它们复杂的蛋白质-蛋白质相互作用，通常是传统小分子药物无法达到的，这使得许多新的细胞因子靶点的临床验证成为可能。尽管取得了成功，但生物药物仍有很大的局限性。挑战包括口服生物利用度差，组织渗透和获得细胞内药物靶点，以及高制造成本。这些限制促使人们努力开发小分子等效物，以复制生物制剂的治疗效果，同时克服其递送能力差和生产成本高的问题。在过去的二十年里，结构生物学、计算建模、疾病生物学、靶向技术和药物化学的进步已经融合在一起，共同使小分子细胞因子调节剂的鉴定和后续进展进入临床开发。展望未来，我们推测该领域未来的药物发现前景，可能会出现针对一系列临床验证的细胞因子（如TSLP和TL1A）的小分子药物。新模式的出现，如细胞外降解剂、口服肽药物以及具有多价和改进递送的下一代生物药物的开发，将加速这种演变。

{"title":"From Large to Small Cytokine Receptor Antagonists","authors":"Pawel Leznicki, Karl Deacon, Bohdan Waszkowycz, John F Unitt","doi":"10.1016/j.slasd.2025.100280","DOIUrl":"10.1016/j.slasd.2025.100280","url":null,"abstract":"<div><div>Here, we review general trends in biological and small molecule cytokine drug discovery, highlighting key learnings from two successful case studies (TNFα and IL-17), which have helped shape our understanding of how best to identify new small molecule cytokine antagonists.</div><div>Over the past three decades, biological drugs have revolutionized the treatment of a wide range of diseases, from oncology to autoimmune diseases. Their ability to modulate extracellular targets, often inaccessible to conventional small molecule drugs due to their complex protein-protein interactions, has enabled the clinical validation of numerous novel cytokine targets.</div><div>Despite their success, biological drugs have significant limitations. Challenges include poor oral bioavailability, tissue penetration and access to intracellular drug targets, as well as high manufacturing costs. These constraints have catalyzed efforts to develop small molecule equivalents that replicate the therapeutic efficacy of biologicals while overcoming their poor delivery and high production costs.</div><div>Over the past twenty years, advances in structural biology, computational modeling, disease biology, hit-finding technologies, and medicinal chemistry have converged to collectively enable the identification and subsequent progression of small molecule cytokine modulators into clinical development.</div><div>Looking ahead, we speculate on the future drug discovery landscape of this field with the likely emergence of small molecules drugs for a range of clinically validated cytokines like TSLP and TL1A. This evolution will be accelerated by the advent of novel modalities like extracellular degraders, oral peptide drugs and the development of next-generation biological drugs with multi-valency and improved delivery.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"38 ","pages":"Article 100280"},"PeriodicalIF":2.7,"publicationDate":"2025-10-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The NLRP3 signaling pathway is a potential target for clinical translation in glioma treatment NLRP3信号通路是胶质瘤治疗中临床翻译的潜在靶点。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-01 DOI: 10.1016/j.slasd.2025.100279

Fengyu Cheng , Yuanyuan Ming , Yunsong Pan , Tianpeng Zhang , Mingxing Li , Rui Zhang , Hui Zhou , Hui Shi

Glioma, particularly glioblastoma (GBM), stands as the most prevalent and formidable tumor in neurosurgery, marked by relatively poor prognosis and high recurrence rates. Despite significant advancements in multimodal therapies—encompassing surgical intervention, radiotherapy, and systemic treatments such as chemotherapy and targeted agents—the overall survival rate for GBM still remains around 5%. In high-grade gliomas, dysregulated inflammatory pathways critically undermine the efficacy of surgical resection and subsequent targeted chemoradiotherapy, which are pivotal for managing these aggressive malignancies. Even with refined surgical techniques and chemotherapeutic mainstays like temozolomide (TMZ), patient prognoses persist in bleak territory, as survival rates languish far below clinical expectations. The NLRP3 inflammatory signaling pathway, extensively studied in this context, drives pathogenesis through allosteric activation and assembly into the NLRP3 inflammasome, catalyzing the maturation of IL-1β and IL-18, thereby triggering cascading inflammatory responses and pyroptosis. The NLRP3 signaling pathway plays a pivotal role in tumor progression and inflammatory responses, and targeted drugs have already entered Phase I clinical trials. Clinically, NLRP3 can serve as a prognostic biomarker, while the combined application of NLRP3-targeted drugs with novel materials and their technical specifics require further exploration. This review explores the NLRP3 pathway’s mechanistic role in glioma progression, reviews cutting-edge clinical research on NLRP3-targeting therapeutics, and evaluates the transformative potential of modulating this pathway in glioma treatment—offering referential insights into its clinical implications and the inflammatory microenvironment’s interplay with tumor dynamics.

神经胶质瘤，特别是胶质母细胞瘤（GBM）是神经外科中最常见和最可怕的肿瘤，其预后较差，复发率高。尽管多模式治疗（包括手术干预、放疗和全身治疗，如化疗和靶向药物）取得了重大进展，但GBM的总生存率仍保持在5%左右。在高级别胶质瘤中，炎症通路失调严重破坏了手术切除和随后的靶向放化疗的疗效，而放化疗对于治疗这些侵袭性恶性肿瘤至关重要。即使有了精细的手术技术和像替莫唑胺（TMZ）这样的主要化疗药物，由于生存率远远低于临床预期，患者的预后仍然很糟糕。在此背景下，NLRP3炎症信号通路被广泛研究，它通过变构激活和组装到NLRP3炎症小体中，催化IL-1β和IL-18的成熟，从而引发级联炎症反应和焦亡，从而驱动发病机制。NLRP3信号通路在肿瘤进展和炎症反应中起着关键作用，靶向药物已经进入I期临床试验。在临床上，NLRP3可以作为一种预后生物标志物，而NLRP3靶向药物与新材料及其技术特性的联合应用还有待进一步探索。本文探讨了NLRP3通路在胶质瘤进展中的机制作用，综述了NLRP3靶向治疗的前沿临床研究，并评估了调节NLRP3通路在胶质瘤治疗中的转化潜力，为其临床意义和炎症微环境与肿瘤动力学的相互作用提供了参考。

{"title":"The NLRP3 signaling pathway is a potential target for clinical translation in glioma treatment","authors":"Fengyu Cheng , Yuanyuan Ming , Yunsong Pan , Tianpeng Zhang , Mingxing Li , Rui Zhang , Hui Zhou , Hui Shi","doi":"10.1016/j.slasd.2025.100279","DOIUrl":"10.1016/j.slasd.2025.100279","url":null,"abstract":"<div><div>Glioma, particularly glioblastoma (GBM), stands as the most prevalent and formidable tumor in neurosurgery, marked by relatively poor prognosis and high recurrence rates. Despite significant advancements in multimodal therapies—encompassing surgical intervention, radiotherapy, and systemic treatments such as chemotherapy and targeted agents—the overall survival rate for GBM still remains around 5%. In high-grade gliomas, dysregulated inflammatory pathways critically undermine the efficacy of surgical resection and subsequent targeted chemoradiotherapy, which are pivotal for managing these aggressive malignancies. Even with refined surgical techniques and chemotherapeutic mainstays like temozolomide (TMZ), patient prognoses persist in bleak territory, as survival rates languish far below clinical expectations. The NLRP3 inflammatory signaling pathway, extensively studied in this context, drives pathogenesis through allosteric activation and assembly into the NLRP3 inflammasome, catalyzing the maturation of IL-1β and IL-18, thereby triggering cascading inflammatory responses and pyroptosis. The NLRP3 signaling pathway plays a pivotal role in tumor progression and inflammatory responses, and targeted drugs have already entered Phase I clinical trials. Clinically, NLRP3 can serve as a prognostic biomarker, while the combined application of NLRP3-targeted drugs with novel materials and their technical specifics require further exploration. This review explores the NLRP3 pathway’s mechanistic role in glioma progression, reviews cutting-edge clinical research on NLRP3-targeting therapeutics, and evaluates the transformative potential of modulating this pathway in glioma treatment—offering referential insights into its clinical implications and the inflammatory microenvironment’s interplay with tumor dynamics.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"36 ","pages":"Article 100279"},"PeriodicalIF":2.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145214683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-01 DOI: 10.1016/j.slasd.2025.100281

Longfei Zhang, Moustafa T. Gabr

Chitinase-3-like 1 (CHI3L1) protein is a secreted glycoprotein involved in various normal physiological processes, while its abnormal elevation is closely associated to carcinogenesis. CHI3L1 recruits and polarizes immune cells into the tumor microenvironment to maintain an immune suppressive environment, and directly stimulate cancer cells to promote their proliferation and migration. Recent studies demonstrated the feasibility of CHI3L1 deletion in cancer treatment on animal models, however, only a limited number of molecular modulators have been developed. To address this gap, a TRIC-based high-throughput screening (HTS) platform was developed, and a library of 5280 molecules was screened. From the screen, 11 hits (hit rate: 0.21 %) were identified as CHI3L1 binders, and 3 compounds (9N05, 11C19, and 3C13) were validated using surface plasmon resonance (SPR). Among them, 9N05 demonstrated the strongest binding affinity towards CHI3L1, and the K_d value was measured as 202.3 ± 76.6 μM. In summary, this proof-of-concept study demonstrates the feasibility of TRIC-based screening for CHI3L1-targeted molecules, and provide a potent tool for the future CHI3L1 molecular modulator development.

几丁质酶-3样1 （CHI3L1）蛋白是一种参与多种正常生理过程的分泌糖蛋白，其异常升高与癌变密切相关。CHI3L1招募和极化免疫细胞进入肿瘤微环境，维持免疫抑制环境，直接刺激癌细胞，促进其增殖和迁移。最近的研究在动物模型上证实了CHI3L1缺失在癌症治疗中的可行性，然而，只有有限数量的分子调节剂被开发出来。为了解决这一空白，开发了基于trics的高通量筛选（HTS）平台，筛选了5280个分子库。筛选得到11个CHI3L1结合物（命中率0.21%），3个化合物（9N05、11C19和3C13）通过表面等离子体共振（SPR）进行了验证。其中9N05对CHI3L1的结合亲和力最强，Kd值为202.3±76.6 μM。总之，这项概念验证研究证明了基于trics筛选CHI3L1靶向分子的可行性，并为未来CHI3L1分子调节剂的开发提供了有力的工具。

{"title":"Temperature-related intensity change (TRIC)-based high throughput screening platform for the discovery of CHI3L1-targeted small molecules","authors":"Longfei Zhang, Moustafa T. Gabr","doi":"10.1016/j.slasd.2025.100281","DOIUrl":"10.1016/j.slasd.2025.100281","url":null,"abstract":"<div><div>Chitinase-3-like 1 (CHI3L1) protein is a secreted glycoprotein involved in various normal physiological processes, while its abnormal elevation is closely associated to carcinogenesis. CHI3L1 recruits and polarizes immune cells into the tumor microenvironment to maintain an immune suppressive environment, and directly stimulate cancer cells to promote their proliferation and migration. Recent studies demonstrated the feasibility of CHI3L1 deletion in cancer treatment on animal models, however, only a limited number of molecular modulators have been developed. To address this gap, a TRIC-based high-throughput screening (HTS) platform was developed, and a library of 5280 molecules was screened. From the screen, 11 hits (hit rate: 0.21 %) were identified as CHI3L1 binders, and 3 compounds (<strong>9N05, 11C19</strong>, and <strong>3C13</strong>) were validated using surface plasmon resonance (SPR). Among them, <strong>9N05</strong> demonstrated the strongest binding affinity towards CHI3L1, and the <em>K</em><sub>d</sub> value was measured as 202.3 ± 76.6 μM. In summary, this proof-of-concept study demonstrates the feasibility of TRIC-based screening for CHI3L1-targeted molecules, and provide a potent tool for the future CHI3L1 molecular modulator development.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"36 ","pages":"Article 100281"},"PeriodicalIF":2.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145269032","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

VOC analysis for rapid, early detection of bacteria, mold, and mycoplasma in cell and tissue cultures 挥发性有机化合物分析快速，早期检测细菌，霉菌，支原体在细胞和组织培养。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-10-01 DOI: 10.1016/j.slasd.2025.100282

Hannah A. Strobel , Manohar P. Bhandari , Eva Borras , Mitchell M. McCartney , Sarah M. Moss , Cristina E Davis , James B. Hoying

Identifying microbial contamination early in cell culture processing can save time, money, and reagents. However, standard strategies for performing contamination testing often take days, leaving production vulnerable to potential spread of bacteria, mold, and mycoplasma. Volatile organic compounds (VOCs) are released by every living organism. These VOCs are increasingly being used to identify cell pathologies, study cell metabolomics, and assess other biological processes. Often, gas chromatography with mass spectrometry (GC–MS) is used to perform these analyses. Here, we utilized gas chromatography with ion mobility spectrometry (GC-IMS) to detect bacteria, mold, and mycoplasma in both cell and tissue cultures. Traditional GC–MS was used to validate the detection of microbes in cell cultures using GC-IMS. In most cases, headspace samples were collected just two hours after inoculations. GC-IMS was able to detect as low as 10 CFU of 5 different industry standard microbes, including both bacteria and mold species. In addition, mycoplasma presence, which is notoriously difficult to test, was detectable at 24 h post-inoculation. GC-IMS is highly sensitive, has a small footprint, requires minimal training, and can provide results in as little as 20 min per sample. Combined, this makes it an ideal strategy for detecting contamination in cell and tissue production workflows. Such rapid detection could save substantial amounts of time, money, and valuable reagents, and reduce risks to patient safety.

在细胞培养过程中早期识别微生物污染可以节省时间、金钱和试剂。然而，执行污染检测的标准策略通常需要几天时间，使生产容易受到细菌，霉菌和支原体的潜在传播。每一个生物体都会释放挥发性有机化合物（VOCs）。这些挥发性有机化合物越来越多地被用于识别细胞病理、研究细胞代谢组学和评估其他生物过程。通常，气相色谱-质谱（GC-MS）用于执行这些分析。在这里，我们利用气相色谱-离子迁移谱法（GC-IMS）检测细胞和组织培养中的细菌、霉菌和支原体。采用传统的气相色谱-质谱法对细胞培养物中微生物的检测进行验证。在大多数情况下，接种后仅两小时采集顶空样本。GC-IMS能够检测到5种不同行业标准微生物的低至10 CFU，包括细菌和霉菌物种。此外，支原体的存在是出了名的难以检测，在接种后24小时可以检测到。GC-IMS高度敏感，占地面积小，只需最少的培训，并且可以在20分钟内提供每个样品的结果。结合起来，这使得它成为检测细胞和组织生产工作流程中污染的理想策略。这种快速检测可以节省大量的时间、金钱和宝贵的试剂，并降低对患者安全的风险。

{"title":"VOC analysis for rapid, early detection of bacteria, mold, and mycoplasma in cell and tissue cultures","authors":"Hannah A. Strobel , Manohar P. Bhandari , Eva Borras , Mitchell M. McCartney , Sarah M. Moss , Cristina E Davis , James B. Hoying","doi":"10.1016/j.slasd.2025.100282","DOIUrl":"10.1016/j.slasd.2025.100282","url":null,"abstract":"<div><div>Identifying microbial contamination early in cell culture processing can save time, money, and reagents. However, standard strategies for performing contamination testing often take days, leaving production vulnerable to potential spread of bacteria, mold, and mycoplasma. Volatile organic compounds (VOCs) are released by every living organism. These VOCs are increasingly being used to identify cell pathologies, study cell metabolomics, and assess other biological processes. Often, gas chromatography with mass spectrometry (GC–MS) is used to perform these analyses. Here, we utilized gas chromatography with ion mobility spectrometry (GC-IMS) to detect bacteria, mold, and mycoplasma in both cell and tissue cultures. Traditional GC–MS was used to validate the detection of microbes in cell cultures using GC-IMS. In most cases, headspace samples were collected just two hours after inoculations. GC-IMS was able to detect as low as 10 CFU of 5 different industry standard microbes, including both bacteria and mold species. In addition, mycoplasma presence, which is notoriously difficult to test, was detectable at 24 h post-inoculation. GC-IMS is highly sensitive, has a small footprint, requires minimal training, and can provide results in as little as 20 min per sample. Combined, this makes it an ideal strategy for detecting contamination in cell and tissue production workflows. Such rapid detection could save substantial amounts of time, money, and valuable reagents, and reduce risks to patient safety.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"36 ","pages":"Article 100282"},"PeriodicalIF":2.7,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145287836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Discovery and validation of small molecule stabilizers of mutant triose phosphate isomerase (TPI) as potential lead candidates for TPI deficiency 突变型磷酸三糖异构酶（TPI）的小分子稳定剂的发现和验证作为TPI缺陷的潜在候选物。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-09-16 DOI: 10.1016/j.slasd.2025.100278

Laura L. Vollmer , Presley Roberts , Samantha L. Eicher , Marta Wołosowicz , Priyal Patel , Joseph R. Figura , Ella R. Donahue , Josh Berkowitz , Dillon Gavlock , Peter Wipf , Matt LaPorte , Steven J. Mullett , Amelle Shillington , Gregg E. Homanics , Michael J. Palladino , Andreas Vogt

Triosephosphate Isomerase deficiency (TPI-Df) is a devastating untreatable childhood metabolic disease resulting in anemia, severe locomotor impairment, and premature death. Numerous single amino acid substitutions in TPI are pathogenic and result in rapidly progressing multisystem disease. Importantly, all known pathogenic TPI-Df mutations result in a protein that retains function, and pathogenesis is known to result from decreased steady state levels of the functioning protein. There are no small molecule therapies for TPI-Df; current treatments are limited to symptomatic support and dietary interventions. We reasoned that a phenotypic screen was most appropriate to capture agents that stabilize mutant TPI and developed a human cellular TPI-Df assay based on a cellular model of the “common” TPI^E105D mutant protein fused with a GFP and a fluorescent ROS biosensor. The assay was implemented for high-content, high-throughput imaging, optimized to full HTS standards, and used to screen a 2,560 compound pilot library and the 220,700 compound NIH MLSMR compound collection to identify candidate compounds for development into small molecule TPI-Df therapies. Hits were validated in dose-response, TPI-Df patient cells, and various orthogonal assays. Limited SAR revealed three promising compound series, which were evaluated for potential mechanisms of action. The lead series had previously been identified as inducers of HIF1 alpha, spawning a novel hypothesis that HIF1 alpha activation might be a potential avenue to treat TPI-Df patients. A lead molecule was moved into preliminary mouse studies to evaluate pharmacokinetics and tissue distribution and was shown to be moderately brain-penetrant. The lead compound is now positioned for target identification studies and efficacy testing in vivo TPI Df models, including a newly validated mouse model.

三磷酸异构酶缺乏症（TPI-Df）是一种毁灭性的无法治愈的儿童代谢性疾病，可导致贫血、严重运动障碍和过早死亡。TPI中的许多单氨基酸取代是致病性的，并导致快速进展的多系统疾病。重要的是，所有已知的致病性TPI-Df突变都会导致一种保留功能的蛋白质，而已知的发病机制是由于功能蛋白的稳态水平下降。目前还没有针对TPI-Df的小分子疗法；目前的治疗仅限于症状支持和饮食干预。我们认为表型筛选最适合捕获稳定突变TPI的药物，并基于“常见”TPIE105D突变蛋白与GFP和荧光ROS生物传感器融合的细胞模型开发了人类细胞TPI- df测定。该方法实现了高含量、高通量成像，优化到完全符合HTS标准，并用于筛选2,560个化合物先导文库和220,700个化合物NIH MLSMR化合物集合，以确定用于开发小分子TPI-Df疗法的候选化合物。hit在剂量-反应、TPI-Df患者细胞和各种正交试验中得到验证。有限的SAR揭示了三个有前途的化合物系列，并对其潜在的作用机制进行了评估。导联系列先前已被确定为HIF1 α的诱导剂，这产生了一个新的假设，即HIF1 α激活可能是治疗TPI-Df患者的潜在途径。一种铅分子被转移到初步的小鼠研究中，以评估药代动力学和组织分布，并显示出适度的脑渗透。目前，该先导化合物已被用于TPI Df模型（包括一种新验证的小鼠模型）的靶点识别研究和有效性测试。

{"title":"Discovery and validation of small molecule stabilizers of mutant triose phosphate isomerase (TPI) as potential lead candidates for TPI deficiency","authors":"Laura L. Vollmer , Presley Roberts , Samantha L. Eicher , Marta Wołosowicz , Priyal Patel , Joseph R. Figura , Ella R. Donahue , Josh Berkowitz , Dillon Gavlock , Peter Wipf , Matt LaPorte , Steven J. Mullett , Amelle Shillington , Gregg E. Homanics , Michael J. Palladino , Andreas Vogt","doi":"10.1016/j.slasd.2025.100278","DOIUrl":"10.1016/j.slasd.2025.100278","url":null,"abstract":"<div><div><em>Triosephosphate Isomerase</em> deficiency (TPI-Df) is a devastating untreatable childhood metabolic disease resulting in anemia, severe locomotor impairment, and premature death. Numerous single amino acid substitutions in <em>TPI</em> are pathogenic and result in rapidly progressing multisystem disease. Importantly, all known pathogenic TPI-Df mutations result in a protein that retains function, and pathogenesis is known to result from decreased steady state levels of the functioning protein. There are no small molecule therapies for TPI-Df; current treatments are limited to symptomatic support and dietary interventions. We reasoned that a phenotypic screen was most appropriate to capture agents that stabilize mutant TPI and developed a human cellular TPI-Df assay based on a cellular model of the “common” TPI<sup>E105D</sup> mutant protein fused with a GFP and a fluorescent ROS biosensor. The assay was implemented for high-content, high-throughput imaging, optimized to full HTS standards, and used to screen a 2,560 compound pilot library and the 220,700 compound NIH MLSMR compound collection to identify candidate compounds for development into small molecule TPI-Df therapies. Hits were validated in dose-response, TPI-Df patient cells, and various orthogonal assays. Limited SAR revealed three promising compound series, which were evaluated for potential mechanisms of action. The lead series had previously been identified as inducers of HIF1 alpha, spawning a novel hypothesis that HIF1 alpha activation might be a potential avenue to treat TPI-Df patients. A lead molecule was moved into preliminary mouse studies to evaluate pharmacokinetics and tissue distribution and was shown to be moderately brain-penetrant. The lead compound is now positioned for target identification studies and efficacy testing in vivo TPI Df models, including a newly validated mouse model.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"36 ","pages":"Article 100278"},"PeriodicalIF":2.7,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145088523","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Integrating AUROC and SSMD for quality control in high-throughput screening assays 整合AUROC和SSMD用于高通量筛选试验的质量控制。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-09-03 DOI: 10.1016/j.slasd.2025.100269

Xiaohua Douglas Zhang

High-throughput screening (HTS) assays are pivotal in modern biomedical research, particularly in drug discovery and functional genomics. Ensuring the quality and reliability of HTS data is critical, especially when dealing with the small sample sizes that are typical in such assays. This study explores the integration of two powerful statistical metrics—Strictly Standardized Mean Difference (SSMD) and Area Under the Receiver Operating Characteristic Curve (AUROC)—for quality control (QC) in HTS. SSMD offers a standardized, interpretable measure of effect size, while AUROC provides a threshold-independent assessment of discriminative power. By establishing the theoretical and empirical relationships between AUROC and SSMD, we demonstrate how these metrics complement each other and enhance QC practices. We provide parametric, semi-parametric, and non-parametric estimation methods, and demonstrate the utility of the integrated framework in real HTS datasets. Our findings support the joint application of SSMD and AUROC as a robust and interpretable approach to improving QC in HTS, particularly under constraints of limited sample sizes of positive and negative controls.

高通量筛选（HTS）分析是现代生物医学研究的关键，特别是在药物发现和功能基因组学方面。确保高温超导数据的质量和可靠性至关重要，特别是在处理此类分析中典型的小样本量时。本研究探讨了两个强大的统计指标-严格标准化平均差（SSMD）和接受者工作特征曲线下面积(AUROC)-在HTS质量控制（QC）中的整合。SSMD提供了一个标准化的、可解释的效应大小测量，而AUROC提供了一个独立于阈值的判别能力评估。通过建立AUROC和SSMD之间的理论和经验关系，我们展示了这些指标如何相互补充并增强质量控制实践。我们提供了参数、半参数和非参数估计方法，并演示了集成框架在实际HTS数据集中的实用性。我们的研究结果支持SSMD和AUROC联合应用作为一种稳健且可解释的方法来改善HTS的质量控制，特别是在阳性和阴性对照的有限样本量的约束下。

{"title":"Integrating AUROC and SSMD for quality control in high-throughput screening assays","authors":"Xiaohua Douglas Zhang","doi":"10.1016/j.slasd.2025.100269","DOIUrl":"10.1016/j.slasd.2025.100269","url":null,"abstract":"<div><div>High-throughput screening (HTS) assays are pivotal in modern biomedical research, particularly in drug discovery and functional genomics. Ensuring the quality and reliability of HTS data is critical, especially when dealing with the small sample sizes that are typical in such assays. This study explores the integration of two powerful statistical metrics—Strictly Standardized Mean Difference (SSMD) and Area Under the Receiver Operating Characteristic Curve (AUROC)—for quality control (QC) in HTS. SSMD offers a standardized, interpretable measure of effect size, while AUROC provides a threshold-independent assessment of discriminative power. By establishing the theoretical and empirical relationships between AUROC and SSMD, we demonstrate how these metrics complement each other and enhance QC practices. We provide parametric, semi-parametric, and non-parametric estimation methods, and demonstrate the utility of the integrated framework in real HTS datasets. Our findings support the joint application of SSMD and AUROC as a robust and interpretable approach to improving QC in HTS, particularly under constraints of limited sample sizes of positive and negative controls.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"36 ","pages":"Article 100269"},"PeriodicalIF":2.7,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145006970","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The role of m6A in lipid metabolism-related diseases m6A在脂质代谢相关疾病中的作用。

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-09-01 DOI: 10.1016/j.slasd.2025.100257

Qianhui Zeng , Yuxuan Yang , Yinquan Zhang , Siwen Jiang

N6-methyladenosine (m⁶A) modification represents one of the most abundant RNA epigenetic modifications in eukaryote organisms. This modification plays a crucial role in various RNA processing events through the actions of m⁶A methyltransferases, demethylases, and recognition proteins, thereby influencing a wide array of biological processes. Notably, the significance of m⁶A modification in lipid metabolism, along with its underlying molecular regulatory mechanisms, is increasingly being elucidated. The regulation of lipid metabolism is intricately linked to the maintenance of energy homeostasis, and disruptions in lipid metabolism are characteristic of numerous diseases, including obesity, non-alcoholic fatty liver disease, and cardiovascular disease. This review comprehensively summarizes the pivotal role and molecular mechanisms of m⁶A modification in diseases related to lipid metabolism, such as obesity, non-alcoholic fatty liver disease, and cardiovascular atherosclerosis. Additionally, it introduces pharmacological agents, plant extracts, and small molecule compounds that target m⁶A regulatory factors. This work provides theoretical references for the development of future therapeutic strategies targeting m⁶A modifications to treat diseases related to lipid metabolism.

n6 -甲基腺苷修饰是真核生物中最丰富的RNA表观遗传修饰之一。这种修饰通过m6A甲基转移酶、去甲基化酶和识别蛋白的作用，在各种RNA加工事件中起着至关重要的作用，从而影响广泛的生物过程。值得注意的是，m6A修饰在脂质代谢中的意义及其潜在的分子调控机制正越来越多地被阐明。脂质代谢的调节与能量稳态的维持有着复杂的联系，脂质代谢的中断是许多疾病的特征，包括肥胖、非酒精性脂肪性肝病和心血管疾病。本文就m6A修饰在肥胖、非酒精性脂肪肝、心血管动脉粥样硬化等脂质代谢相关疾病中的关键作用及分子机制进行综述。此外，它还介绍了针对m6A调节因子的药理学试剂、植物提取物和小分子化合物。这项工作为未来针对m6A修饰治疗脂质代谢相关疾病的治疗策略的制定提供了理论参考。

{"title":"The role of m6A in lipid metabolism-related diseases","authors":"Qianhui Zeng , Yuxuan Yang , Yinquan Zhang , Siwen Jiang","doi":"10.1016/j.slasd.2025.100257","DOIUrl":"10.1016/j.slasd.2025.100257","url":null,"abstract":"<div><div>N6-methyladenosine (m<sup>6</sup>A) modification represents one of the most abundant RNA epigenetic modifications in eukaryote organisms. This modification plays a crucial role in various RNA processing events through the actions of m<sup>6</sup>A methyltransferases, demethylases, and recognition proteins, thereby influencing a wide array of biological processes. Notably, the significance of m<sup>6</sup>A modification in lipid metabolism, along with its underlying molecular regulatory mechanisms, is increasingly being elucidated. The regulation of lipid metabolism is intricately linked to the maintenance of energy homeostasis, and disruptions in lipid metabolism are characteristic of numerous diseases, including obesity, non-alcoholic fatty liver disease, and cardiovascular disease. This review comprehensively summarizes the pivotal role and molecular mechanisms of m<sup>6</sup>A modification in diseases related to lipid metabolism, such as obesity, non-alcoholic fatty liver disease, and cardiovascular atherosclerosis. Additionally, it introduces pharmacological agents, plant extracts, and small molecule compounds that target m<sup>6</sup>A regulatory factors. This work provides theoretical references for the development of future therapeutic strategies targeting m<sup>6</sup>A modifications to treat diseases related to lipid metabolism.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100257"},"PeriodicalIF":2.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144805449","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cytotoxic Profiling of Korea Chemical Bank Diversity Library 韩国化学银行多样性库的细胞毒性分析

IF 2.7 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

SLAS Discovery

Pub Date : 2025-09-01 DOI: 10.1016/j.slasd.2025.100267

Jinhee Kim , Kwang-Eun Choi , Yuno Lee , Daeyoung Jeong , Hyun Young Kim , Jung-In Lee , Heeyeong Cho , Nam-Chul Cho

Cytotoxicity profiling of screening libraries is a critical component in early-stage drug discovery to identify compounds with undesirable toxic effects. Here, we report the cytotoxicity profiling of the Korea Chemical Bank (KCB) diversity library, comprising 7040 compounds curated via virtual screening, clustering, and druggability assessment. A subset of 5181 compounds was randomly selected and screened using the WST-1 assay in five mammalian cell lines (HEK293, HFL1, HepG2, NIH3T3, and CHOK1) at concentrations of 30 µM and 10 µM, following 24 h and 48 h incubation periods. Cytotoxic compounds were defined as those exhibiting >50 % inhibition at 30 µM after 48 h. A total of 17 compounds showed consistent cytotoxicity across all five cell lines. Comparative analysis of physicochemical properties revealed that cytotoxic compounds exhibited higher lipophilicity (ALogP/ LogD) and the number of aromatic rings (AR) relative to non-cytotoxic compounds. These results indicate that the majority of the KCB diversity library comprises non-cytotoxic compounds, reflecting effective pre-filtering of toxic physicochemical properties during library design.

筛选文库的细胞毒性分析是早期药物发现中识别具有不良毒性作用的化合物的关键组成部分。在这里，我们报告了韩国化学银行（KCB）多样性文库的细胞毒性分析，该文库包括7040种化合物，通过虚拟筛选、聚类和药物性评估。在5种哺乳动物细胞系（HEK293、HFL1、HepG2、NIH3T3和CHOK1）中，随机选择5181个化合物，在浓度为30µM和10µM的条件下，经过24 h和48 h的孵育，使用WST-1试验进行筛选。细胞毒性化合物被定义为48小时后在30µM下表现出50%的抑制作用。共有17种化合物在所有5种细胞系中表现出一致的细胞毒性。理化性质对比分析表明，细胞毒性化合物的亲脂性（ALogP/ LogD）和芳香环数（AR）均高于非细胞毒性化合物。这些结果表明，大多数KCB多样性文库包含非细胞毒性化合物，反映了在文库设计过程中对毒性物理化学性质进行了有效的预过滤。

{"title":"Cytotoxic Profiling of Korea Chemical Bank Diversity Library","authors":"Jinhee Kim , Kwang-Eun Choi , Yuno Lee , Daeyoung Jeong , Hyun Young Kim , Jung-In Lee , Heeyeong Cho , Nam-Chul Cho","doi":"10.1016/j.slasd.2025.100267","DOIUrl":"10.1016/j.slasd.2025.100267","url":null,"abstract":"<div><div>Cytotoxicity profiling of screening libraries is a critical component in early-stage drug discovery to identify compounds with undesirable toxic effects. Here, we report the cytotoxicity profiling of the Korea Chemical Bank (KCB) diversity library, comprising 7040 compounds curated via virtual screening, clustering, and druggability assessment. A subset of 5181 compounds was randomly selected and screened using the WST-1 assay in five mammalian cell lines (HEK293, HFL1, HepG2, NIH3T3, and CHOK1) at concentrations of 30 µM and 10 µM, following 24 h and 48 h incubation periods. Cytotoxic compounds were defined as those exhibiting >50 % inhibition at 30 µM after 48 h. A total of 17 compounds showed consistent cytotoxicity across all five cell lines. Comparative analysis of physicochemical properties revealed that cytotoxic compounds exhibited higher lipophilicity (ALogP/ LogD) and the number of aromatic rings (AR) relative to non-cytotoxic compounds. These results indicate that the majority of the KCB diversity library comprises non-cytotoxic compounds, reflecting effective pre-filtering of toxic physicochemical properties during library design.</div></div>","PeriodicalId":21764,"journal":{"name":"SLAS Discovery","volume":"35 ","pages":"Article 100267"},"PeriodicalIF":2.7,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144921842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0