IspD (2-C-Methyl-D-erythritol 4-Phosphate Cytidyltransferase), an Essential and Druggable Antimalarial Target

Overview

Journal ACS Infect Dis

Specialties Infectious Diseases
Microbiology
Pharmacology

Date 2016 Jan 20

PMID 26783558

Citations 33

Authors

Leah S Imlay

Christopher M Armstrong

Mary Clare Masters

Ting Li

Kathryn E Price

Rachel L Edwards

Katherine M Mann

Lucy X Li

Christina L Stallings

Neil G Berry

Paul M ONeill

Audrey R Odom

Affiliations

Soon will be listed here.

Abstract

As resistance to current therapies spreads, novel antimalarials are urgently needed. In this work, we examine the potential for therapeutic intervention via the targeting of IspD (2-C-methyl-D-erythritol 4-phosphate cytidyltransferase), the second dedicated enzyme of the essential methylerythritol phosphate (MEP) pathway for isoprenoid biosynthesis. Enzymes of this pathway represent promising therapeutic targets because the pathway is not present in humans. The Malaria Box compound, MMV008138, inhibits growth, and PfIspD has been proposed as a candidate intracellular target. We find that PfIspD is the sole intracellular target of MMV008138 and characterize the mode of inhibition and target-based resistance, providing chemical validation of this target. Additionally, we find that the genetic locus is refractory to disruption in malaria parasites, providing independent genetic validation for efforts targeting this enzyme. This work provides compelling support for IspD as a druggable target for the development of additional, much-needed antimalarial agents.

Citing Articles

Dissecting apicoplast functions through continuous cultivation of Toxoplasma gondii devoid of the organelle.

Chen M, Koszti S, Bonavoglia A, Maco B, von Rohr O, Peng H Nat Commun. 2025; 16(1):2095.

PMID: 40025025 PMC: 11873192. DOI: 10.1038/s41467-025-57302-x.

Targeting IspD for Anti-infective and Herbicide Development: Exploring Its Role, Mechanism, and Structural Insights.

Willocx D, Diamanti E, Hirsch A J Med Chem. 2025; 68(2):886-901.

PMID: 39749898 PMC: 11770629. DOI: 10.1021/acs.jmedchem.4c01146.

Fragment Discovery by X-Ray Crystallographic Screening Targeting the CTP Binding Site of Pseudomonas Aeruginosa IspD.

Willocx D, DAuria L, Walsh D, Scherer H, Alhayek A, Hamed M Angew Chem Int Ed Engl. 2024; 64(6):e202414615.

PMID: 39676054 PMC: 11796317. DOI: 10.1002/anie.202414615.

Targeting IspD in the Methyl-d-erythritol Phosphate Pathway: Urea-Based Compounds with Nanomolar Potency on Target and Low-Micromolar Whole-Cell Activity.

Willocx D, Bizzarri L, Alhayek A, Kannan D, Bravo P, Illarionov B J Med Chem. 2024; 67(19):17070-17086.

PMID: 39303294 PMC: 11472328. DOI: 10.1021/acs.jmedchem.4c00212.

Antimalarial drug discovery: progress and approaches.

Siqueira-Neto J, Wicht K, Chibale K, Burrows J, Fidock D, Winzeler E Nat Rev Drug Discov. 2023; 22(10):807-826.

PMID: 37652975 PMC: 10543600. DOI: 10.1038/s41573-023-00772-9.

References

Borrmann S, Lundgren I, Oyakhirome S, Impouma B, Matsiegui P, Adegnika A . Fosmidomycin plus clindamycin for treatment of pediatric patients aged 1 to 14 years with Plasmodium falciparum malaria. Antimicrob Agents Chemother. 2006; 50(8):2713-8. PMC: 1538678. DOI: 10.1128/AAC.00392-06. View

Spangenberg T, Burrows J, Kowalczyk P, McDonald S, Wells T, Willis P . The open access malaria box: a drug discovery catalyst for neglected diseases. PLoS One. 2013; 8(6):e62906. PMC: 3684613. DOI: 10.1371/journal.pone.0062906. View

Ashley E, Dhorda M, Fairhurst R, Amaratunga C, Lim P, Suon S . Spread of artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2014; 371(5):411-23. PMC: 4143591. DOI: 10.1056/NEJMoa1314981. View

Dondorp A, Nosten F, Yi P, Das D, Phyo A, Tarning J . Artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2009; 361(5):455-67. PMC: 3495232. DOI: 10.1056/NEJMoa0808859. View

Kelley L, Sternberg M . Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc. 2009; 4(3):363-71. DOI: 10.1038/nprot.2009.2. View

Plouffe D, Brinker A, McNamara C, Henson K, Kato N, Kuhen K . In silico activity profiling reveals the mechanism of action of antimalarials discovered in a high-throughput screen. Proc Natl Acad Sci U S A. 2008; 105(26):9059-64. PMC: 2440361. DOI: 10.1073/pnas.0802982105. View

Huang D, Zhou T, Lafleur K, Nevado C, Caflisch A . Kinase selectivity potential for inhibitors targeting the ATP binding site: a network analysis. Bioinformatics. 2009; 26(2):198-204. DOI: 10.1093/bioinformatics/btp650. View

Vriend G . WHAT IF: a molecular modeling and drug design program. J Mol Graph. 1990; 8(1):52-6, 29. DOI: 10.1016/0263-7855(90)80070-v. View

Borrmann S, Adegnika A, Matsiegui P, Issifou S, Schindler A, Mawili-Mboumba D . Fosmidomycin-clindamycin for Plasmodium falciparum Infections in African children. J Infect Dis. 2004; 189(5):901-8. DOI: 10.1086/381785. View

10.

Tsang A, Seidle H, Jawaid S, Zhou W, Smith C, Couch R . Francisella tularensis 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase: kinetic characterization and phosphoregulation. PLoS One. 2011; 6(6):e20884. PMC: 3111433. DOI: 10.1371/journal.pone.0020884. View

11.

Baer R . With the ends in sight: images from the BRCA1 tumor suppressor. Nat Struct Biol. 2001; 8(10):822-4. DOI: 10.1038/nsb1001-822. View

12.

Zhang B, Watts K, Hodge D, Kemp L, Hunstad D, Hicks L . A second target of the antimalarial and antibacterial agent fosmidomycin revealed by cellular metabolic profiling. Biochemistry. 2011; 50(17):3570-7. PMC: 3082593. DOI: 10.1021/bi200113y. View

13.

Wiley J, Merino E, Krai P, McLean K, Tripathi A, Vega-Rodriguez J . Isoprenoid precursor biosynthesis is the essential metabolic role of the apicoplast during gametocytogenesis in Plasmodium falciparum. Eukaryot Cell. 2014; 14(2):128-39. PMC: 4311923. DOI: 10.1128/EC.00198-14. View

14.

Vaughan A, ONeill M, Tarun A, Camargo N, Phuong T, Aly A . Type II fatty acid synthesis is essential only for malaria parasite late liver stage development. Cell Microbiol. 2008; 11(3):506-20. PMC: 2688669. DOI: 10.1111/j.1462-5822.2008.01270.x. View

15.

Alexandrov A, Vignali M, LaCount D, Quartley E, de Vries C, De Rosa D . A facile method for high-throughput co-expression of protein pairs. Mol Cell Proteomics. 2004; 3(9):934-8. DOI: 10.1074/mcp.T400008-MCP200. View

16.

Ruangweerayut R, Looareesuwan S, Hutchinson D, Chauemung A, Banmairuroi V, Na-Bangchang K . Assessment of the pharmacokinetics and dynamics of two combination regimens of fosmidomycin-clindamycin in patients with acute uncomplicated falciparum malaria. Malar J. 2008; 7:225. PMC: 2600645. DOI: 10.1186/1475-2875-7-225. View

17.

Gamo F, Sanz L, Vidal J, de Cozar C, Alvarez E, Lavandera J . Thousands of chemical starting points for antimalarial lead identification. Nature. 2010; 465(7296):305-10. DOI: 10.1038/nature09107. View

18.

TRAGER W, Jensen J . Human malaria parasites in continuous culture. Science. 1976; 193(4254):673-5. DOI: 10.1126/science.781840. View

19.

Odom A, Van Voorhis W . Functional genetic analysis of the Plasmodium falciparum deoxyxylulose 5-phosphate reductoisomerase gene. Mol Biochem Parasitol. 2009; 170(2):108-11. PMC: 2814890. DOI: 10.1016/j.molbiopara.2009.12.001. View

20.

Bernal C, Palacin C, Boronat A, Imperial S . A colorimetric assay for the determination of 4-diphosphocytidyl-2-C-methyl-D-erythritol 4-phosphate synthase activity. Anal Biochem. 2005; 337(1):55-61. DOI: 10.1016/j.ab.2004.10.011. View