Novel peptidomimetic MMP-9 inhibitors as potential anti-epileptogenic therapeutics

• Autorzy: Khomiak D. , Bertran A. , Rejmak E. , Kaczmarek L.
• Języki publikacji: EN

Abstrakty

EN

INTRODUCTION: Epilepsy is the most widespread neurological disorder (prevalence – 50 million). The recent discoveries suggest that remodelling of the brain extracellular matrix, executed by extracellularly operating proteases, may play a fundamental role in the pathogenesis of epilepsy. One of them, MMP-9 has been particularly linked to epileptogenesis and the functional involvement of MMP-9 in kainic acid and pentylenetetrazole-kindling models of temporal lobe epilepsy demonstrated. AIM(S): Considering the inhibition MMP-9 as a promising therapeutic strategy, MMP-9 Inhibitors of peptidomimetic nature IPR-X, IPR-Y and IPR-Z were developed at Iproteos and initial conjoint testing of these compounds has been performed. METHOD(S): Compounds ability to penetrate blood‑brain barrier was analyzed by PAMPA (Parallel artificial membrane permeability assay. Stability to degradation in liver and cytotoxicity were tested by exposing to rat liver microsomes and by MTT assay, respectively. Selectivity and potency of IPR‑X‑Z were pre‑estimated by fluorescent assay using DQ‑gelatin and recombinant MMP‑9. Finally, the inhibitors’ ability to inhibit cleavage of of MMP-9 substrate – Nectin-3 was tested in primary hippocampal cell culture upon 50 μM glutamate stimulation. RESULTS: PAMPA assay studies yielded in 7.7–13.3% penetration percentage for artificial blood-brain barrier. The values of intrinsic clearance for IPR-X-Z determined by stability in liver microsomes assay ranged from 33 to 46 µL/min/mg protein, indicating moderate degradation. Compounds proved to be non-toxic in MTT cytotoxicity assay, except of IPR-Z at the highest concentration tested (200 µM). IC50 values from DQ‑gelatinase assay were shown to be 4–10 µM. In hippocampal cell cultures compounds inhibited MMP-9 dependent nectin-3 cleavage with 60% of total band intensity remaining on western blot. CONCLUSIONS: The obtained results confirm that IPR‑X, IPR‑Y and IPR‑Z are specific and potent towards MMP-9 and could be further tested in animal models of epileptogenesis. FINANCIAL SUPPORT: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 642881.

• Wydawca: -
• Czasopismo: Acta Neurobiologiae Experimentalis
• Rocznik: 2017
• Tom: 77
• Numer: Suppl.1
• Opis fizyczny: p.57

Twórcy

autor

Khomiak D.

• Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland

autor

Bertran A.

• Iproteos, Barcelona Science Park, Barcelona, Spain

autor

Rejmak E.

• Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland

autor

Kaczmarek L.

• Department of Neurophysiology, Nencki Institute of Experimental Biology, Warsaw, Poland