Inhibition of PrP^Scformation in scrapie infected N2a cells by 5,7,8-trimethyl-3,4-dihydro-2H-1,4-benzoxazine derivatives

Abstract

Prion diseases are fatal, neurodegenerative diseases characterized by the structural conversion of the normal, cellular prion protein, PrP (C) into an abnormally structured, aggregated and partially protease-resistant isoform, termed PrP (Sc) . Although substantial research has been directed toward development of therapeutics targeting prions, there is still no curative treatment for the disease. Benzoxazines are bicyclic heterocyclic compounds possessing several pharmaceutically important properties, including neuroprotection and reactive oxygen species scavenging. In an effort to identify novel inhibitors of prion formation, several 5,7,8-trimethyl-1,4-benzoxazine derivatives were evaluated in vitro for their effectiveness on the expression levels of normal PrP (C) and its conversion to the abnormal isoforms of PrP (Sc) in a scrapie-infected cell culture model. The most potent compound was 2-(4-methoxyphenyl)-5,7,8-trimethyl-3,4-dihydro-2H-1,4-benzoxazine, with a diminishing effect on the formation of PrP (Sc) , thus establishing a class of compounds with a promising therapeutic use against prion diseases.

Carnosic Acid and Carnosol Display Antioxidant and Anti-Prion Properties in In Vitro and Cell-Free Models of Prion Diseases

“…In this study, we used 22L-scrapie-infected mouse neuroblastoma cells (N2a22L), a well-established in the field [25,26,44] in vitro prion disease model, to evaluate the antiprion effects of Carnosic acid and Carnosol. We first assessed the potential toxic effects of the compounds on the cell lines used in the study by incubating the cells for 48 h in the presence of different concentrations of the compounds and estimating the survival rates using the MTT assay.…”

“…Several compounds have been tested for anti-prion activity, including larger molecules such as pentosan polysulfate [18,19], suramin [19,20], amphotericin B [20][21][22], Congo red [22,23], and dendritic polyamines [23,24], and have been shown to possess anti-prion properties. Among small molecules, bis-acridines [24,25], 5,7,8-trimethyl-3,4-dihydro-2H-1,4-benzoxazine derivatives [25,26], polyphenols, phenothiazines, antihistamines, statins, some antimalarial agents [18][19][20][21][22][23][24][25][26][27], indole-3-glyoxamides [28,29], pyridyl hydrazones [29,30], 2-aminothiazoles [30,31], a group of phenethyl piperidines [31,32], and glycodendrimers [32,33] have been shown to inhibit PrP Sc formation in vitro. However, there is still a need for effective compounds when administered before and/or at the onset of neurological symptoms.…”

Humoral response in experimental autoimmune encephalomyelitis targets neural precursor cells in the central nervous system of naive rodents

“…Murine Neuro2a (N2a) cell line was kindly provided by Prof. T. Sklaviadis, Pharmacy department of A.U.TH (Thessaloniki, Greece), and cultured according to standard protocol [ 20 ].…”

Πειραματική Μελέτη Αντιγονικότητας Των Προγονικών Κυττάρων Του Κεντρικού Νευρικού Συστήματος Σε C57bl/6 Ποντικούς

“…Η διάλυση όλων των δεύτερων αντισωμάτων έγινε σε TBS 1x. Αντισώματα που χρησιμοποιήθηκαν στα πειράματα ανοσοφθορισμού.Τα κύτταρα νευροβλαστώματος από μύες, Ν2a (Neuro2a), είναι έτοιμη κυτταρική σειρά που μας προσέφερε το εργαστήριο Φαρμακογνωσίας-Φαρμακολογίας του τμήματος Φαρμακευτικής Α.Π.Θ., αποθηκευμένη στους -80 ο C και καλλιεργήθηκε βάση συγκεκριμένου πρωτοκόλλου[235]. Πιο συγκεκριμένα, το cryovial με τα κύτταρα απόψυχθηκε με ιδιαίτερη προσοχή, καθώς περιέχει DMSO, το οποίο είναι κρυοπροστατευτικό, αλλά σε θερμοκρασία δωματίου καθίσταται τοξικό για τα κύτταρα.…”