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Pharmacokinetics and Metabolism of 2-Aminothiazoles with Antiprion Activity in Mice

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Abstract

Purpose

To discover drugs lowering PrPSc in prion-infected cultured neuronal cells that achieve high concentrations in brain to test in mouse models of prion disease and then treat people with these fatal diseases.

Methods

We tested 2-AMT analogs for EC50 and PK after a 40 mg/kg single dose and 40–210 mg/kg/day doses for 3 days. We calculated plasma and brain AUC, ratio of AUC/EC50 after dosing. We reasoned that compounds with high AUC/EC50 ratios should be good candidates going forward.

Results

We evaluated 27 2-AMTs in single-dose and 10 in 3-day PK studies, of which IND24 and IND81 were selected for testing in mouse models of prion disease. They had high concentrations in brain after oral dosing. Absolute bioavailability ranged from 27–40%. AUC/EC50 ratios after 3 days were >100 (total) and 48–113 (unbound). Stability in liver microsomes ranged from 30–>60 min. Ring hydroxylated metabolites were observed in microsomes. Neither was a substrate for the MDR1 transporter.

Conclusions

IND24 and IND81 are active in vitro and show high AUC/EC50 ratios (total and unbound) in plasma and brain. These will be evaluated in mouse models of prion disease.

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Abbreviations

2-AMT:

2-aminothiazole scaffold

AUC:

area under the drug concentration time curve

C3-day :

drug concentration after 3 days of dosing

CJD:

Creutzfeldt-Jakob disease

Cl:

clearance (total, intrinsic, hepatic, or otherwise)

Cmax :

maximum drug concentration

dpi:

days postinoculation with prions

EC50 :

potency; drug concentration producing 50% of the maximal effect

FaSSIF:

fasted-state simulated intestinal fluid

HTS:

high-throughput screening

IV:

intravenous

MDCK-MDR1:

Madin Darby canine kidney cells transfected with MDR1 gene

MDR1:

multidrug resistance protein 1, ATP-binding cassette sub-family B member 1

MIC:

minimum inhibitory concentration

P-gp:

p-glycoprotein

PK:

pharmacokinetics

PO:

oral

PrPC :

benign normally occurring prion protein on cell surface or inside cell

PrPSc :

abnormal, misfolded, pathogenic form of PrPC

RML:

Rocky Mountain Laboratory

SAR:

structure-activity relationship

ScN2a-cl3:

scrapie (RML)-infected neuroblastoma cells that overexpress PrPC

V:

volume of distribution (steady-state or otherwise)

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Acknowledgments AND DISCLOSURES

The authors thank Ms. Ana Serban, Ms. Julia Becker, and Mr. Frederic Letessier for D13 and D18 antibodies; Mr. Phillip Benner and the staff of the Hunter’s Point animal facility for expert animal studies; Dr. Sina Ghaemmaghami for many helpful discussions; and Ms. Hang Nguyen for editorial assistance. This work was supported by grants from the National Institutes of Health (AG002132, AG010770, AG031220, and AG021601) as well as by gifts from the Sherman Fairchild Foundation, Rainwater Charitable Foundation, Lincy Foundation, Fight for Mike Homer Program, and Robert Galvin. MPJ is a consultant to Schrodinger LLC.

Author information

Authors and Affiliations

  1. Institute for Neurodegenerative Diseases, University of California, San Francisco, California, USA

    B. Michael Silber, Satish Rao, Kimberly L. Fife, Kurt Giles, Yevgeniy Freyman, Manuel Elepano, Joel R. Gever, Zhe Li & Stanley B. Prusiner

  2. Department of Neurology, University of California, San Francisco, California, USA

    B. Michael Silber, Satish Rao, Kurt Giles, Joel R. Gever, Zhe Li & Stanley B. Prusiner

  3. Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, California, USA

    B. Michael Silber, Kimberly L. Fife, Zhe Li, Yong Huang & Leslie Z. Benet

  4. Small Molecule Discovery Center, University of California, San Francisco, California, USA

    Alejandra Gallardo-Godoy & Adam R. Renslo

  5. Department of Pharmaceutical Chemistry, University of California, San Francisco, California, USA

    Adam R. Renslo & Matthew P. Jacobson

  6. Pfizer Global Research & Development, La Jolla, California, USA

    Deepak K. Dalvie

  7. 675 Nelson Rising Ln, Room 318, San Francisco, California, 94143-0518, USA

    Stanley B. Prusiner

Authors
  1. B. Michael Silber
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  2. Satish Rao
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  3. Kimberly L. Fife
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  10. Joel R. Gever
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  15. Stanley B. Prusiner
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Corresponding author

Correspondence to Stanley B. Prusiner.

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Silber, B.M., Rao, S., Fife, K.L. et al. Pharmacokinetics and Metabolism of 2-Aminothiazoles with Antiprion Activity in Mice. Pharm Res 30, 932–950 (2013). https://doi.org/10.1007/s11095-012-0912-4

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  • DOI: https://doi.org/10.1007/s11095-012-0912-4

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