Skip to main content
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems
  • Log in
  • My alerts
  • My Cart

Main menu

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • AAC Podcast
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
  • ASM
    • Antimicrobial Agents and Chemotherapy
    • Applied and Environmental Microbiology
    • Clinical Microbiology Reviews
    • Clinical and Vaccine Immunology
    • EcoSal Plus
    • Eukaryotic Cell
    • Infection and Immunity
    • Journal of Bacteriology
    • Journal of Clinical Microbiology
    • Journal of Microbiology & Biology Education
    • Journal of Virology
    • mBio
    • Microbiology and Molecular Biology Reviews
    • Microbiology Resource Announcements
    • Microbiology Spectrum
    • Molecular and Cellular Biology
    • mSphere
    • mSystems

User menu

  • Log in
  • My alerts
  • My Cart

Search

  • Advanced search
Antimicrobial Agents and Chemotherapy
publisher-logosite-logo

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Accepted Manuscripts
    • COVID-19 Special Collection
    • Archive
    • Minireviews
  • For Authors
    • Submit a Manuscript
    • Scope
    • Editorial Policy
    • Submission, Review, & Publication Processes
    • Organization and Format
    • Errata, Author Corrections, Retractions
    • Illustrations and Tables
    • Nomenclature
    • Abbreviations and Conventions
    • Publication Fees
    • Ethics Resources and Policies
  • About the Journal
    • About AAC
    • Editor in Chief
    • Editorial Board
    • For Reviewers
    • For the Media
    • For Librarians
    • For Advertisers
    • Alerts
    • AAC Podcast
    • RSS
    • FAQ
  • Subscribe
    • Members
    • Institutions
Chemistry; Biosynthesis

Hematin Polymerization Assay as a High-Throughput Screen for Identification of New Antimalarial Pharmacophores

Yae Kurosawa, Arnulf Dorn, Michiko Kitsuji-Shirane, Hisao Shimada, Tomoko Satoh, Hugues Matile, Werner Hofheinz, Raffaello Masciadri, Manfred Kansy, Robert G. Ridley
Yae Kurosawa
Department of Pharmaceutical Screening, Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa Prefecture 247,
New Ceramics Department, Asahi Optical Co., Ltd., Itabashi-ku, Tokyo 174-8639,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Arnulf Dorn
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michiko Kitsuji-Shirane
Department of Pharmaceutical Screening, Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa Prefecture 247,
Medical Institute of Bioregulation, Department of Molecular and Cellular Biology, Kyushu University, 3-1-1 Maidaishi, Higashi-ku Fukuoka, Fukuoka 812-8582, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hisao Shimada
Department of Pharmaceutical Screening, Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa Prefecture 247,
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tomoko Satoh
Department of Pharmaceutical Screening, Nippon Roche Research Center, 200 Kajiwara, Kamakura, Kanagawa Prefecture 247,
Screening Technology, Bayer Yakuhin Research Center Kyoto, 6-5-1-3 Kunimidai, Soraku-gun, Kyoto, Japan, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Hugues Matile
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Werner Hofheinz
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Raffaello Masciadri
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Manfred Kansy
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Robert G. Ridley
Pharma Division, Preclinical Research, F. Hoffmann-La Roche Ltd., CH-4070 Basel, and
Drug Discovery Research, WHO/TDR, World Health Organization, CH-1211 Geneva 27, Switzerland
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1128/AAC.44.10.2638-2644.2000
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Fig. 1.
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    Screening cascade for chemical cocktails, SPECS compounds, and microbial broths. Schematically shown are the procedures and selection criteria leading from primary screening to in vitro testing and the start of a medicinal chemistry program. First, the inhibitory activity should be above a defined value (e.g., ≥40% inhibition at 50 μM) and must be confirmed; second, the structure must be of interest (acceptable molecular weight; lipophilicity) and have a potential for chemical variations; third, the identified hit would, preferentially, have in vitro activity.

  • Fig. 2.
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    (A) The monoquinoline-derived hypothesis superimposed on chloroquine. (B) Fit of the monoquinoline-derived hypothesis for Ro 06-8463, which was found by 3-D database screening (activity for the resistant strain K1, 0.3 μM; activity for the sensitive strain NF54, 0.026 μM). Feature definitions: blue, hydrophobic; green, hydrogen bond acceptor; cyan, hydrogen bond donor; red, positive ionizable.

Tables

  • Figures
  • Table 1.

    Effects of substances possibly interfering with the hematin polymerization reaction

    Class and compoundConcn% InhibitionIC50Concn tolerated
    Cations and anions
     NaCl100 mM10
     KCl100 mM10
     NH4Cl100 mM14
     MgCl2100 mM16
     CaCl2100 mM−15
     Na2HPO4100 mM26
     NaH2PO4100 mM14
    Detergents
     Tween 801%880.14%0.003%
     Triton X-1001%37
     SDS1%46
    Chelating agents
     EDTA10 mM−19
     EGTA10 mM−10
    Tannins
     Catechin100 μg/ml11
     Gallic acid100 μg/ml−43
     Tannic acid100 μg/ml10034 μg/ml3 μg/ml
    Pigments
     Chlorophyll a100 μg/ml33
     Chlorophyll b100 μg/ml28
     Melanin100 μg/ml40
    Saponin (saikosaponin A)100 μg/ml−9
    Antibiotics
     Daunomycin100 μg/ml−27
     Amphotericin B100 μg/ml4
     Penicillic acid100 μg/ml−19
     Actinomycin D100 μg/ml37
    SH-group reacting agents
     DTT100 μg/ml895.1 μg/ml1 μg/ml
     Cysteine100 μg/ml−13
     GSH100 μg/ml41
    Fatty acids
     Stearic acid100 μg/ml11
     Oleic acid100 μg/ml30
     Linoleic acid100 μg/ml−15
    Proteases
     Actinase E10 μg/ml−13
     S peptidase10 μg/ml−29
     Proteinase K10 μg/ml−6
    Organic solvents
     MeOH10%−29
     EtOH10%−28
     Acetonitrile10%−40
     DMSO10%−37
  • Table 2.

    Results of tests for inhibition of hematin polymerization

    Chemical source of compoundsNo. of compounds in primary screeningNo. (%) of hits in:No. (%) of active single compounds
    Primary screeningReassay
    Roche cocktails11,887268 (2.3)69 (0.6)38 (0.3)
    NCC9716 (16)16 (16)5 (5.2)
    SPECS15,76036 (0.2)13 (0.1)
    Combinatorial chemistry libraries1,3800
    Microbial broths
     Actinomycetes6,00023 (0.4)3 (0.1)0
     Fungi6,32042 (0.7)17 (0.3)10 (0.2)
     Bacteria2,32013 (0.6)4 (0.2)1
  • Table 3.

    Classes of compounds identified as inhibitors of the hematin polymerization reaction in HTS

    Compound classStructureIC50 (μM) in hematin polymerization assayIC50 (μM) against P. falciparum growth in culture% Inhibition ofP. berghei growth in vivoaIC50 (μM) in cytotoxicity assayb
    K1NF54
    Chloroquine (Ro 01-6014)Embedded Image800.450.0125100NT
    Triarylcarbinol (Ro 06-9075)300.971.1698.44.66
    Piperazine (Ro 10-3428)<1000.210.27Inactive4.45
    Miscellaneous (Ro 14-3955)241.932.7Inactive63.3
    Benzophenone (Ro 22-8014)241.321.7888NT
    Imide (Ro 20-1120)131616NTNT
    Hydrazide (Ro 04-4410)1915.415.4InactiveNT
    Indole (Ro 90-2341)1428.528.5NTNT
    Isoxazole (Ro 90-0987)1228.528.5NTNT
    • ↵a Mice were treated p.o. with four doses of each test compound at 100 mg/kg of body weight.

    • ↵b With HeLa cells. NT, not tested.

  • Table 4.

    Selection of the most active compounds found by 3-D database screening

    Compound structureRoche no.IC50 (μM) in hematin polymerization assayIC50 (μM) against P. falciparum growth in culture
    NF54K1
    Embedded ImageRo 06-8463280.0260.3
    Ro 06-9075301.160.97
    Ro 10-3428<1000.270.21
    Ro 10-89951000.140.29
    Ro 47-3191 NTa0.460.3
    • ↵a NT, not tested.

PreviousNext
Back to top
Download PDF
Citation Tools
Hematin Polymerization Assay as a High-Throughput Screen for Identification of New Antimalarial Pharmacophores
Yae Kurosawa, Arnulf Dorn, Michiko Kitsuji-Shirane, Hisao Shimada, Tomoko Satoh, Hugues Matile, Werner Hofheinz, Raffaello Masciadri, Manfred Kansy, Robert G. Ridley
Antimicrobial Agents and Chemotherapy Oct 2000, 44 (10) 2638-2644; DOI: 10.1128/AAC.44.10.2638-2644.2000

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Print

Alerts
Sign In to Email Alerts with your Email Address
Email

Thank you for sharing this Antimicrobial Agents and Chemotherapy article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Hematin Polymerization Assay as a High-Throughput Screen for Identification of New Antimalarial Pharmacophores
(Your Name) has forwarded a page to you from Antimicrobial Agents and Chemotherapy
(Your Name) thought you would be interested in this article in Antimicrobial Agents and Chemotherapy.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Hematin Polymerization Assay as a High-Throughput Screen for Identification of New Antimalarial Pharmacophores
Yae Kurosawa, Arnulf Dorn, Michiko Kitsuji-Shirane, Hisao Shimada, Tomoko Satoh, Hugues Matile, Werner Hofheinz, Raffaello Masciadri, Manfred Kansy, Robert G. Ridley
Antimicrobial Agents and Chemotherapy Oct 2000, 44 (10) 2638-2644; DOI: 10.1128/AAC.44.10.2638-2644.2000
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Top
  • Article
    • ABSTRACT
    • MATERIALS AND METHODS
    • RESULTS AND DISCUSSION
    • ACKNOWLEDGMENTS
    • FOOTNOTES
    • REFERENCES
  • Figures & Data
  • Info & Metrics
  • PDF

KEYWORDS

antimalarials
Hemin

Related Articles

Cited By...

About

  • About AAC
  • Editor in Chief
  • Editorial Board
  • Policies
  • For Reviewers
  • For the Media
  • For Librarians
  • For Advertisers
  • Alerts
  • AAC Podcast
  • RSS
  • FAQ
  • Permissions
  • Journal Announcements

Authors

  • ASM Author Center
  • Submit a Manuscript
  • Article Types
  • Ethics
  • Contact Us

Follow #AACJournal

@ASMicrobiology

       

ASM Journals

ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.

About ASM | Contact Us | Press Room

 

ASM is a member of

Scientific Society Publisher Alliance

 

American Society for Microbiology
1752 N St. NW
Washington, DC 20036
Phone: (202) 737-3600

Copyright © 2021 American Society for Microbiology | Privacy Policy | Website feedback

Print ISSN: 0066-4804; Online ISSN: 1098-6596