PZ-2891 is an orally bioavailable, brain-penetrant pantothenate kinase (PANK) modulator, with an IC50 of 1.3 nM for hPANK3.

PANK1β (human):40.2±4.4 nM (IC50), PANK2 (human):0.7±0.08 nM (IC50), PANK3 (human):1.3±0.2 nM (IC50)

Oral administration of PZ-2891 increases CoA levels in mouse liver and brain.?A knockout mouse model of brain CoA deficiency exhibited weight loss, severe locomotor impairment and early death.?Knockout mice on PZ-2891 therapy gain weight, and have improved locomotor activity and life span establishing pantazines as novel therapeutics for the treatment of PKAN.

C57Bl6/J mice (8-week-old) ?pantothenic acid-supplemented diet (1000 ppm) for 2 weeks prior to the experiment.?The mice were maintained at room temperature 72° ± 2 °F, humidity 50 ± 10% and a 14 h light /10 h dark cycle with the dark cycle starting at 18:00 h. Water was supplied ad libitum.?The mice were randomized into the treatment arms to achieve a normal weight distribution.?PZ-2891 was formulated in 30% Captisol, and was administered by oral gavage at 12 h intervals for 5 doses.?The mice were euthanized and tissues harvested 4 h after the last dose.?The tissue samples were used for total CoA, pantothenate and pantazine determinations.?Blood was collected from euthanized animals, plasma or serum was prepared and stored frozen until analysis.?Organs, including liver, forebrain, and hindbrain were quickly excised from euthanized animals and immediately flash frozen in liquid N2 or immersed in RNAlater overnight prior to freezing.?Forebrain and hindbrain regions were identified.?Total CoA was determined using 20–50 mg of tissue (liver, forebrain or hindbrain) homogenized in 2 ml of 1 mM KOH.?The pH was adjusted to 12.0 with 0.25 M KOH, and incubated at 55 °C for 2 h.?The pH of the sample was adjusted to 8.0, and the samples were derivatized with monobromobimane (mBBr) and analyzed by HPLC equipped with a fluorescence detector