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In-Vivo and Ex-Vivo Brain Uptake Studies of Peptidomimetic Neurolysin Activators in Healthy and Stroke Animals

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Abstract

Purpose

Neurolysin (Nln) is a peptidase that functions to preserve the brain following ischemic stroke by hydrolyzing various neuropeptides. Nln activation has emerged as an attractive drug discovery target for treatment of ischemic stroke. Among first-in-class peptidomimetic Nln activators, we selected three lead compounds (9d, 10c, 11a) for quantitative pharmacokinetic analysis to provide valuable information for subsequent preclinical development.

Methods

Pharmacokinetic profile of these compounds was studied in healthy and ischemic stroke-induced mice after bolus intravenous administration. Brain concentration and brain uptake clearance (Kin) was calculated from single time point analysis. The inter-relationship between LogP with in-vitro and in-vivo permeability was studied to determine CNS penetration. Brain slice uptake method was used to study tissue binding, whereas P-gp-mediated transport was evaluated to understand the potential brain efflux of these compounds.

Results

According to calculated parameters, all three compounds showed a detectable amount in the brain after intravenous administration at 4 mg/kg; however, 11a had the highest brain concentration and brain uptake clearance. A strong correlation was documented between in-vitro and in-vivo permeability data. The efflux ratio of 10c was ~6-fold higher compared to 11a and correlated well with its lower Kin value. In experimental stroke animals, the Kin of 11a was significantly higher in ischemic vs. contralateral and intact hemispheres, though it remained below its A50 value required to activate Nln.

Conclusions

Collectively, these preclinical pharmacokinetic studies reveal promising BBB permeability of 11a and indicate that it can serve as an excellent lead for developing improved drug-like Nln activators.

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Acknowledgments and Disclosures

Authors declare no conflict of interest.

Funding

This project was supported by NIH grant R01NS106879 to P.C.T., V.T.K. and T.J.A.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Jerry. H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, 1300 Coulter St, Amarillo, TX, 79106, USA

    Saeideh Nozohouri, Shiva Hadi Esfahani, Behnam Noorani, Heidi Villalba, Yashwardhan Ghanwatkar, Yong Zhang, Ulrich Bickel, Vardan T. Karamyan & Thomas J. Abbruscato

  2. Center for Blood Brain Barrier Research, Jerry. H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, TX, USA

    Saeideh Nozohouri, Shiva Hadi Esfahani, Behnam Noorani, Heidi Villalba, Yashwardhan Ghanwatkar, Yong Zhang, Ulrich Bickel, Vardan T. Karamyan & Thomas J. Abbruscato

  3. Department of Pharmaceutical Sciences, College of Pharmacy, University of Nebraska Medical Center (UNMC), Omaha, NE, 68106, USA

    Md. Shafikur Rahman & Paul C. Trippier

  4. UNMC Center for Drug Discovery, University of Nebraska Medical Center, Omaha, NE, 68106, USA

    Paul C. Trippier

  5. Office of Sciences, Jerry. H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, TX, 79106, USA

    Dhaval Patel

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  1. Saeideh Nozohouri
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  2. Shiva Hadi Esfahani
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  3. Behnam Noorani
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  5. Heidi Villalba
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  7. Md. Shafikur Rahman
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  11. Vardan T. Karamyan
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  12. Thomas J. Abbruscato
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Correspondence to Thomas J. Abbruscato.

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Nozohouri, S., Esfahani, S.H., Noorani, B. et al. In-Vivo and Ex-Vivo Brain Uptake Studies of Peptidomimetic Neurolysin Activators in Healthy and Stroke Animals. Pharm Res 39, 1587–1598 (2022). https://doi.org/10.1007/s11095-022-03218-w

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  • DOI: https://doi.org/10.1007/s11095-022-03218-w

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