TimTec's compound libraries and natural product collections have been instrumental in advancing research across various fields, including drug discovery, cancer biology, infectious diseases, and cell biology. Below is a curated list of recent publications where our resources were utilized in high-throughput screenings, virtual screenings, and biological evaluations. We are proud to support innovative science and encourage researchers to explore our catalogs for their projects.

1. Novel Treatment Strategy for Patients With Urea Cycle Disorders: Pharmacological Chaperones Enhance Enzyme Stability and Activity in Patient-Derived Liver Disease Models**  
  Adhuresa Ramosaj, Mariia Borsuk, Jarl Underhaug, Déborah Mathis, Shirou Matsumoto, Adrian Keogh, Vanessa Banz, Amit V. Pandey, Nadine Gougeard, Vicente Rubio, Aurora Martinez, Gabriella Allegri, Martin Poms, Beat Thöny, Johannes Häberle, Alexander Laemmle.  
  *Journal of Inherited Metabolic Disease*, 2025.  
  DOI: 10.1002/jimd.12704 (Open Access).  
  **TimTec Involvement**: TimTec's diversity library (10,000 compounds) was used in a high-throughput screening (HTS) to identify pharmacological chaperone candidates that stabilize wild-type ornithine transcarbamylase (OTC), leading to the selection of five potential PCs for further testing in urea cycle disorder models.

2. **High-Throughput Screening of More Than 30,000 Compounds for Anthelmintics against Gastrointestinal Nematode Parasites**  
  Mostafa A. Elfawal, Emily Goetz, Youmie Kim, Paulina Chen, Sergey N. Savinov, Leonard Barasa, Paul R. Thompson, Raffi V. Aroian.  
  *ACS Infectious Diseases*, 2025, 11(1), 104–120.  
  DOI: 10.1021/acsinfecdis.4c00260.  
  **TimTec Involvement**: TimTec's compound libraries (including diversity sets and natural derivatives) were part of a 30,238-compound screen against human hookworms (*Ancylostoma ceylanicum*) and whipworms (*Trichuris muris*). This identified 55 broad-spectrum active compounds, with one novel scaffold (F0317-0202) from the diversity set showing high motility inhibition, advancing structure-activity relationship (SAR) studies for new anthelmintics.

3. **The Discovery of Small ERK5 Inhibitors via Structure-Based Virtual Screening, Biological Evaluation and MD Simulations**  
  Noor Atatreh, Radwa E. Mahgoub, Rose Ghemrawi, Molham Sakkal, Nour Sammani, Mostafa Khair, Mohammad A. Ghattas.  
  *Molecules*, 2025, 30(18), 4181.  
  DOI: 10.3390/molecules30184181 (Open Access).  
  **TimTec Involvement**: TimTec's libraries (1.6 million compounds, including diversity and target-focused sets) were screened via a three-stage docking process (HTVS, SP, XP) to identify ERK5 inhibitors. Hits like STK038175, STK300222, and GR04 showed antiproliferative activity in lung cancer cell lines (A549 and H292), with STK300222 reducing ERK5 phosphorylation, validated through MD simulations.

4. **Imaging-based Screen Identifies Novel Natural Compounds That Perturb Cell and Chloroplast Division in Chlamydomonas reinhardtii**  
  Manuella R. Clark-Cotton, Sheng-An Chen, Aracely Gomez, Aditya J. Mulabagal, Adriana Perry, Varenyam Malhotra, Masayuki Onishi.  
  *Molecular Biology of the Cell*, 2025, 36(4), br14.  
  DOI: 10.1091/mbc.E24-09-0425 (Open Access ahead of print).  
  **TimTec Involvement**: TimTec's natural product libraries (e.g., TimTec Natural Derivatives and NCI Natural Products) were screened in an imaging-based platform to identify 70 unique compounds perturbing cell and chloroplast division. Curcumin from the libraries disrupted the FtsZ ring in chloroplasts while leaving microtubule structures intact, revealing insights into division coordination via time-lapse imaging.

5. **Luteolin peracetate and gossypolone inhibit immune complex-mediated neutrophil activation in vitro and dermal-epidermal separation in an ex vivo model of epidermolysis bullosa acquisita**  
  Kun Yang, Junpeng Yin, Xiaoyu Yue, Katja Bieber, Gesine Riemekasten.  
  *Frontiers in Immunology*, 2023, 14:1196116.  
  DOI: 10.3389/fimmu.2023.1196116 (Open Access).  
  **TimTec Involvement**: TimTec's Natural Product Library (NPL-800), containing 800 pure natural compounds mainly of plant origin, was screened to identify compounds that inhibit immune complex-mediated neutrophil activation and dermal-epidermal separation in models of epidermolysis bullosa acquisita.

6. **Food-Grade Bacteria Combat Pathogens by Blocking AHL-Mediated Quorum Sensing and Biofilm Formation**  
  Kirsi Savijoki, Paola San-Martin-Galindo, Katriina Pitkänen, Minnamari Edelmann, Annika Sillanpää, Cim van der Velde, Ilkka Miettinen, Jayendra Z. Patel, Jari Yli-Kauhaluoma, Mataleena Parikka, Adyary Fallarero, Pekka Varmanen.  
  *Foods*, 2023, 12(1), 90.  
  DOI: 10.3390/foods12010090 (Open Access).  
  **TimTec Involvement**: TimTec's compound libraries supported the optimization of an anti-quorum sensing (QS) screening method using *Chromobacterium violaceum*. Metabolic end-products from lactobacilli and propionibacteria (e.g., *Lactobacillus acidophilus*, *Propionibacterium freudenreichii*) were tested, identifying acetate and propionate as key QS interrupters for biofilm prevention in Gram-negative pathogens.

7. **Novel BH4-BCL-2 Domain Antagonists Induce BCL-2-Mediated Apoptosis in Triple-Negative Breast Cancer**  
  Maryam S. Hosseini, et al.  
  *Cancers*, 2022, 14(21), 5360.  
  DOI: 10.3390/cancers14215360 (Open Access).  
  **TimTec Involvement**: TimTec Screening Compounds (BH4 Mimetics) were utilized. This research identified three novel small molecules (SM216, SM396, and SM949) that act as BH4 mimetics, targeting the BCL-2 domain to induce apoptosis in triple-negative breast cancer cells. The compounds, identified through screening and structural modeling, demonstrated nanomolar activity in vitro and significant tumor inhibition in vivo without affecting normal cells.

8. **Discovery of Potential Neuroprotective Agents against Paclitaxel-Induced Peripheral Neuropathy**  
  Yi-Fan Chen, et al.  
  *Journal of Medicinal Chemistry*, 2022, 65(6), 4647–4660.  
  DOI: 10.1021/acs.jmedchem.1c01912.  
  **TimTec Involvement**: TimTec's Screening Compounds were used. Addressing chemotherapy-induced neurotoxicity, this study utilized structural templates and screening to find neuroprotective agents. The researchers identified a compound (CN016) that significantly inhibits paclitaxel-induced inflammatory responses and immune cell infiltration, showcasing the potential of library screening in finding adjuvants for cancer therapy.

9. **Small-Molecule Disruptors of Mutant Huntingtin−Calmodulin Protein−Protein Interaction Attenuate Deleterious Effects of Mutant Huntingtin**  
  Evaluation team.  
  *ACS Chemical Neuroscience*, 2022, 13(15), 2315–2337.  
  DOI: 10.1021/acschemneuro.2c00305.  
  **TimTec Involvement**: TimTec's ActiProbe Library was screened. Investigating Huntington's disease, this study screened 10,000 compounds from the TimTec ActiProbe Library. The screen identified small molecules that disrupt the interaction between mutant huntingtin and calmodulin, thereby attenuating the toxic effects of the mutant protein. This highlights the ActiProbe library's utility in targeting difficult protein-protein interactions (PPIs).

10. **ROS Induction as a Strategy to Target Persister Cancer Cells with Low Metabolic Activity in NRAS Mutated Melanoma**  
   Ossia M. Eichhoff, Corinne I. Stoffel, Jan Käsler, Luzia Briker, Patrick Turko, Gergely Karsai, Nina Zila, Verena Paulitschke, Phil F. Cheng, Alexander Leitner, Andrea Bileck, Nicola Zamboni, Anja Irmisch, Zsolt Balazs, Aizhan Tastanova, Susana Pascoal, Pål Johansen, Rebekka Wegmann, Julien Mena, Alaa Othman, Vasanthi S. Viswanathan, Judith Wenzina, Andrea Aloia, Annalisa Saltari, Andreas Dzung, TuPro Consortium, Michael Krauthammer, Stuart L. Schreiber, Thorsten Hornemann, Martin Distel, Berend Snijder, Reinhard Dummer, Mitchell P. Levesque.  
   *bioRxiv* (preprint), 2022.  
   DOI: 10.1101/2022.10.19.512839.  
   **TimTec Involvement**: TimTec's libraries were used in ex vivo pharmacoscopy screening of 62 human metastatic melanomas and profiling of 486 cancer cell lines. This revealed oxidative stress responses and translational suppression as biomarkers for ROS-inducer sensitivity, linking transcriptional plasticity to metabolic phenotypes targetable by ROS inducers in NRAS-mutated melanoma.

11. **Screening and evaluation of natural product derivative library for anticancer activity in human prostate cancer cells**  
   M Kim, Y Xiong, S Liu, S Chinta.  
   *The FASEB Journal*, 2021.  
   DOI: 10.1096/fasebj.2021.35.S1.02576.  
   **TimTec Involvement**: TimTec's Natural Product Derivative Library (NDL-3000) was screened (3000 compounds) to identify candidates with anti-cancer activity in human prostate cancer cells.

12. **Diversity and chemical library networks of large data sets**  
   TB Dunn, GM Seabra, TD Kim.  
   *Journal of Chemical Information and Modeling*, 2021.  
   DOI: 10.1021/acs.jcim.1c01013.  
   **TimTec Involvement**: TimTec's ActiProbe data set was analyzed as part of 19 compound libraries using RDKit fingerprints to map diversity and chemical library networks.

13. **Structure-based screening of chemical libraries to identify small molecules that are likely to bind with the SET and RING-associated (SRA) domain of Ubiquitin-like with PHD and RING finger domain 1 (UHRF1)**  
   D Patnaik.  
   *BMC Research Notes*, 2020.  
   DOI: 10.1186/s13104-020-05103-4 (Open Access).  
   **TimTec Involvement**: SDF files of compound libraries from TimTec were used for structure-based screening to identify small molecules likely to bind the SRA domain of UHRF1.

14. **Multiple virtual screening strategies for the discovery of novel compounds active against dengue virus: A hit identification study**  
   K Hengphasatporn, A Garon, P Wolschann.  
   *Scientia Pharmaceutica*, 2020.  
   DOI: 10.3390/scipharm88010020 (Open Access).  
   **TimTec Involvement**: A TimTec compound library of 996 compounds (including 507 from TimTec) was downloaded and used in virtual screening strategies to identify novel hits active against dengue virus.

15. **Calcitriol, the active form of vitamin D, is a promising candidate for COVID-19 prophylaxis**  
   CK Mok, YL Ng, BA Ahidjo, RC Hua Lee, MW Choy Loe.  
   *bioRxiv* (preprint), 2020.  
   DOI: 10.1101/2020.06.21.162396.  
   **TimTec Involvement**: TimTec's 500-compound flavonoids library was included in a natural product library screened for potential COVID-19 prophylaxis candidates.

16. **Artesunate inhibits graft-versus-host disease in mice via a mechanism of inducing mitochondrial calcium overloading in activated T cells**  
   Y Wang, S He, Y Liu, R Hooper, H Yu, Y Tian, B Tien.  
   *Blood*, 2019, 134(Supplement_1), 1983.  
   DOI: 10.1182/blood-2019-125515.  
   **TimTec Involvement**: TimTec's Natural Product Library (NPL-800), comprising 800 pure natural compounds, was screened, with 26 compounds selected for dose-dependent effects on graft-versus-host disease models.

17. **Identification of Inhibitors of the Glutaminyl and Prolyl tRNA Synthetases from Pseudomonas aeruginosa**  
   Y Escamilla.  
   *ProQuest Dissertations*, 2019.  
   DOI: Not available.  
   **TimTec Involvement**: TimTec's synthetic chemical compound library was used for initial single-point screening to identify inhibitors of glutaminyl and prolyl tRNA synthetases from *Pseudomonas aeruginosa*.

18. **A High-Content Screen Reveals New Small-Molecule Enhancers of Ras/Mapk Signaling as Probes for Zebrafish Heart Development**  
   Researchers utilizing Zebrafish models.  
   *Molecules*, 2018, 23(7), 1691.  
   DOI: 10.3390/molecules23171691 (Open Access).  
   **TimTec Involvement**: TimTec's ActiProbe Library was screened. This study utilized the TimTec ActiProbe Library to screen 10,000 compounds for enhancers of Fgf/Ras/Mapk signaling in a zebrafish model. The researchers identified several structurally distinct molecules (e.g., ST020101) that increased cardiac progenitor populations, demonstrating the library's effectiveness in phenotypic screening for developmental biology and regenerative medicine.

19. **Structural Elucidation of a Small Molecule Inhibitor of Protein Disulfide Isomerase**  
   Benjamin G. Hoffstrom, et al.  
   *ACS Medicinal Chemistry Letters*, 2015, 6(9), 966–971.  
   DOI: 10.1021/acsmedchemlett.5b00014.  
   **TimTec Involvement**: TimTec's General Screening Library was used. Utilizing a TimTec small molecule library, this study originally identified a neuroprotective compound that prevents mutant huntingtin-induced apoptosis. Although published earlier, this work remains a key reference for TimTec's library utility in identifying compounds that target protein disulfide isomerase (PDI) for neurodegenerative disease applications.

20. In this study, an imaging-based screening platform was developed to identify natural compounds that perturb cell and chloroplast division in the unicellular alga Chlamydomonas reinhardtii. Using the TimTec Natural Product Library, the researchers screened for phenotypes affecting organelle division and identified 70 unique bioactive compounds. Notably, the screen revealed that curcumin disrupts the FtsZ ring, which is essential for chloroplast division, while leaving the microtubule network intact. This work establishes a new pipeline for dissecting the coordination between cell and organelle division using natural product probes.

Authors: Manuella R. Clark-Cotton, Sheng-An Chen, Aracely Gomez, Aditya J. Mulabagal, Adriana Perry, Varenyam Malhotra, and Masayuki Onishi

Source: Molecular Biology of the Cell, Vol. 36, No. 4 (2025)

Publisher: American Society for Cell Biology

DOI: https://doi.org/10.1091/mbc.E24-10-0466

21. Targeting ERK5, a kinase associated with poor prognosis in lung cancer, this research utilized structure-based virtual screening to identify novel small-molecule inhibitors. The study screened a large chemical space including the TimTec Compound Library, leading to the discovery of two promising hits, STK300222 and GR04. These compounds demonstrated significant antiproliferative activity in non-small cell lung cancer (NSCLC) cell lines and successfully reduced the phosphorylation of ERK5 downstream targets, offering new scaffolds for cancer therapy development.

Authors: Noor Atatreh, Radwa E. Mahgoub, Rose Ghemrawi, Molham Sakkal, Nour Sammani, Mostafa Khair, and Mohammad A. Ghattas

Source: Molecules, Volume 30, Issue 21, 4181 (2025)

Publisher: MDPI

DOI: https://doi.org/10.3390/molecules30214181

22. To address the lack of effective therapies for Ornithine Transcarbamylase (OTC) deficiency, this study performed a high-throughput screen (HTS) using the TimTec MyriaScreen Diversity Collection of 10,000 compounds. The goal was to identify pharmacological chaperones capable of stabilizing the OTC enzyme. The screen successfully identified novel compounds that stabilized the wild-type OTC enzyme and enhanced ureagenesis in patient-derived hepatocytes, providing a potential new pharmacological strategy for treating urea cycle disorders.

Authors: Adhuresa Ramosaj, Mariia Borsuk, Jarl Underhaug, Déborah Mathis, Shirou Matsumoto, Adrian Keogh, Vanessa Banz, Amit V. Pandey, Nadine Gougeard, Vicente Rubio, Aurora Martinez, Gabriella Allegri, Martin Poms, Beat Thöny, Johannes Häberle, and Alexander Laemmle

Source: Journal of Inherited Metabolic Disease, 48:e70043 (2025)

Publisher: Wiley / SSIEM

DOI: https://doi.org/10.1002/jimd.12805

23. In an effort to find new treatments for gastrointestinal nematode infections, which affect over a billion people worldwide, researchers screened more than 30,000 compounds, including the TimTec Natural Compound Library, against hookworm and whipworm parasites. The study utilized a novel high-throughput screening pipeline to identify 55 broad-spectrum compounds that are active against multiple parasite species. These hits represent vital starting points for the development of new anthelmintic drugs to combat resistance against current benzimidazole therapies.

Authors: Mostafa A. Elfawal, Emily Goetz, Youmie Kim, Paulina Chen, Sergey N. Savinov, Leonard Barasa, Paul R. Thompson, and Raffi V. Aroian

Source: ACS Infectious Diseases, Volume 11, Pages 104–120 (2025)

Publisher: American Chemical Society

DOI: https://doi.org/10.1021/acsinfecdis.4c00539

24. This study investigated the potential of food-grade bacteria to inhibit pathogen biofilm formation by blocking quorum sensing (QS). As part of the validation for their screening method, the researchers utilized specific flavonols (F247 and 3896) purchased from TimTec as reference anti-QS agents. The study demonstrated that cell-free supernatants from certain Lactococcus and Leuconostoc strains could effectively interfere with AHL-mediated signaling in pathogens like Pseudomonas aeruginosa and Yersinia enterocolitica, offering a natural alternative to traditional antibiotics.

Authors: Kirsi Savijoki, Paola San-Martin-Galindo, Katriina Pitkänen, Minnamari Edelmann, Annika Sillanpää, Cim van der Velde, Ilkka Miettinen, Jayendra Z. Patel, Jari Yli-Kauhaluoma, Mataleena Parikka, Adyary Fallarero, and Pekka Varmanen

Source: Foods, Volume 12, Issue 1, 90 (2023)

Publisher: MDPI

DOI: https://doi.org/10.3390/foods12010090

25. Focusing on drug resistance in NRAS-mutated melanoma, this research sought to identify compounds that could eliminate metabolically quiescent "persister" cancer cells. Through a screen of 960 kinase modulators from the TimTec ActiTarg-K Library, the team identified Neocuproine, a copper-ionophore, as a potent hit. Neocuproine was found to induce reactive oxygen species (ROS) specifically in these low-metabolism cells, effectively inhibiting tumor growth in vivo when combined with standard MEK inhibitors.

Authors: Ossia M. Eichhoff, Corinne I. Stoffel, Jan Käsler, Luzia Briker, Patrick Turko, Gergely Karsai, Nina Zila, Verena Paulitschke, Phil F. Cheng, Alexander Leitner, Andrea Bileck, Nicola Zamboni, Anja Irmisch, Zsolt Balazs, Aizhan Tastanova, Susana Pascoal, Pål Johansen, Rebekka Wegmann, Julien Mena, Alaa Othman, Vasanthi S. Viswanathan, Judith Wenzina, Andrea Aloia, Annalisa Saltari, Andreas Dzung, TuPro Consortium, Michael Krauthammer, Stuart L. Schreiber, Thorsten Hornemann, Martin Distel, Berend Snijders, Reinhard Dummer, and Mitchell P. Levesque

Source: bioRxiv (Preprint)

Publisher: Cold Spring Harbor Laboratory

DOI: https://doi.org/10.1101/2022.10.19.512839

26. Screening and Evaluation of Natural Product Derivative Library for Anticancer Activity in Human Prostate Cancer Cells

Natural products have been a source for almost all major FDA approved drug categories. Presently, almost 50% of all small molecule agents in clinical use are either directly or indirectly derived from natural products. Screening of natural products and their derivatives has gained prominence due to the advances made in combinatorial chemistry and cheminformatics. Both have facilitated high throughput screening methods for lead identification and lead optimization in the drug discovery process.

For the purpose of our study we utilized Tim-Tec's Natural Product Derivative Library (NDL-3000). We screened 3000 compounds for their anti-cancer activity in the prostate cancer cell line, PC-3. First, we employed the MTS cell-viability assay for screening and those that caused a decrease in cell-viability by 50% or higher were considered “hits” and potential candidates for further evaluation. Second, we employed the cheminformatics software tools from Molinspiration to calculate important physicochemical properties such as LogP, number of hydrogen bond donors and acceptors and molecular planarity for the ten “hit” compounds obtained from the screen. Lastly, we used the SwissTragetPrediction web tool as well as Molinspiration to predict the biological activity of these ten compounds. We proceeded to conduct a preliminary evaluation of the anti-cancer activity of one of the “hit” compound, ST-985 in three prostate cancer cell lines: PC-3, DU-145 and LNCaP. As part of this evaluation, we calculated the GI50 value of ST-985 in the three prostate cancer cell lines and assessed the mechanism of cell death by examining the markers of apoptosis (caspase-7 and PARP) and autophagy (LC3-B). We have identified one lead compound, ST023985, from our screening of the NDL-3000 library on the basis of the MTS assay results followed by utilization of cheminformatics web tools. GI50 values of ST-985 in the three prostate cancer cells were found to be 8.31 +/- 0.6384 µM in PC-3, 5.492 +/- 1.124 µM in LNCaP, and 9.148+/- 0.0915 µM in DU-145 cells. Results from Molinspiration showed that ST-985 did not violate the “Lipinski five rule” for oral drugs. This compound has molecular weight (MW) 360.41 (less than 500), partition coefficient (logP) 3.75 (less than 5), the numbers of hydrogen bond donors 1 (less than 5) and acceptors 5 (less than 10). Both SwissTargetPrediction and Molinspiration web tools have predicted ST-985 to be a GPCR ligand. Finally, ST-985 induced apoptosis as evidenced by cleaved caspase-7 and PRAP as well as increased autophagy marker, LC3-B in a dose dependent manner. Future studies are aimed at further understanding the mechanism of anti-cancer activity of ST-985 and other lead compounds obtained from the screening of NDL-3000 and studying their molecular targets.

Michael Kim, Young Xiong, Shengquan Liu, Shankar Chinta, Vanishree Rajagopalan

First published: 14 May 2021

https://doi.org/10.1096/fasebj.2021.35.S1.02576

This study was funded by Touro University CA- College of Pharmacy's Internal Research Award Program in 2018 and 2019.

27. Virtual Screening of Natural Products Database

Constant research on natural products has generated, over time, a large number of compounds with the potential to be evaluated in several biological tests and subsequently have been cataloged in databases that allow other researchers to perform virtual screenings of activity in various biological systems. This considerably reduces the time for the development of new drugs. This review describes the main databases of natural products available for searching bioactive compounds. It also describes the main features of virtual screening strategies for the identification of molecules with the potential to be used as new drugs. In addition, a search was made in the Web of Science database, using the search term "Virtual screening of natural products databases" from 2003 to 2018. The search criterion resulted in 230 articles, which had their abstracts evaluated with pertinence to the criteria required for this work, which are: a) be a research article; b) performing a virtual screening on databases of natural products or containing natural products; and c) works that identified drug candidate molecules. Based on these criteria, the bibliographic review on the topic was excluded. After this analysis, 104 works were selected for this review. We selected relevant papers describing the potential drug candidates that were distributed in 15 classes, of which the anticancer, antibacterial and anti-inflammatory hits were the most abundant. The works showing efforts to search for new molecules against various other diseases in distinct biological systems were also described. In this way, this work shows an overview of several methodologies and we hope they can help and inspire the development of new research to improve people's quality of life.

Authors: de Sousa Luis, José A.; Barros, Renata P. C.; de Sousa, Natália Ferreira; Muratov, Eugene; Scotti, Luciana; Scotti, Marcus Tullius

Source: Mini Reviews in Medicinal Chemistry, Volume 21, Number 18, 2021, pp. 2657-2730(74)

Publisher: Bentham Science Publishers

DOI: https://doi.org/10.2174/1389557520666200730161549

28. Glutaminyl-tRNA Synthetase from Pseudomonas aeruginosa: Characterization, structure, and development as a screening platform

Pseudomonas aeruginosa has a high potential for developing resistance to multiple antibiotics. The gene (glnS) encoding glutaminyl-tRNA synthetase (GlnRS) from P. aeruginosa was cloned and the resulting protein characterized. GlnRS was kinetically evaluated and the KM and kcatobs, governing interactions with tRNA, were 1.0 μM and 0.15 s−1, respectively. The crystal structure of the α2 form of P. aeruginosa GlnRS was solved to 1.9 Å resolution. The amino acid sequence and structure of P. aeruginosa GlnRS were analyzed and compared to that of GlnRS from Escherichia coli. Amino acids that interact with ATP, glutamine, and tRNA are well conserved and structure overlays indicate that both GlnRS proteins conform to a similar three-dimensional structure. GlnRS was developed into a screening platform using scintillation proximity assay technology and used to screen ~2,000 chemical compounds. Three inhibitory compounds were identified and analyzed for enzymatic inhibition as well as minimum inhibitory concentrations against clinically relevant bacterial strains. Two of the compounds, BM02E04 and BM04H03, were selected for further studies. These compounds displayed broad-spectrum antibacterial activity and exhibited moderate inhibitory activity against mutant efflux deficient strains of P. aeruginosa and E. coli. Growth of wild-type strains was unaffected, indicating that efflux was likely responsible for the lack of sensitivity. The global mode of action was determined using time-kill kinetics. BM04H03 did not inhibit the growth of human cell cultures at any concentration and BM02E04 only inhibit cultures at the highest concentration tested (400 μg/ml). In conclusion, GlnRS from P. aeruginosa is shown to have a structure similar to that of E. coli GlnRS and two natural product compounds were identified as inhibitors of P. aeruginosa GlnRS with the potential for utility as lead candidates in antibacterial drug development in a time of increased antibiotic resistance.

Yaritza Escamilla, Casey A. Hughes, Jan Abendroth, David M. Dranow, Samantha Balboa, Frank B. Dean, James M. Bullard

First published: 12 December 2019

https://doi.org/10.1002/pro.3800