Pridopidine (ACR16)

Cell experiment:

Conditionally immortalized mouse striatal knock-in cells expressing endogenous levels of wild-type (STHdh7/7) or mHtt (STHdh111/111) are used. Different concentrations of Pridopidine (100, 150, 200 and 300 μM) are tested to investigate the anti-apoptotic effect of the molecule on immortalized cells cultured in serum-free medium at 39°C for six hours. In NE100 experiments, cells are pre-incubated with the compound (10 μM) for 2 hrs before culturing them in apoptotic conditions. At the end of each treatment, cells are collected and incubated with FITC-conjugated Annexin V. Fluorescence Activated Cell Sorting (FACS) analysis is performed[2].

Animal experiment:

Rats[1]Sprague Dawley (SD) male rats (n=6 per group) are treated daily by oral gavage with Pridopidine at a dose of 60 mg/kg or vehicle (water) over the course of 10 days. On day 10, 90 min following last drug/water administration, brains are removed, and quickly rinsed with cold physiological saline. The striatum of the left hemisphere is gently extracted and immediately immerged in 1000 µL of RNAlater Solution in pre-labelled polypropylene vials and stored at 4°C overnight (to allow the solution to thoroughly penetrate the tissue), then moved to -20°C until analysis. RNA is isolated from the striatum of each rat and analysed[1].Mice[2]All in vivo experiments are conducted in R6/2 transgenic mice expressing exon 1 of human Htt with approximately 160±10 (CAG) repeats and manifesting first symptoms around week 7, and in wild-type (WT) littermates maintained on the B6CBA strain. Animals are housed singly and maintained under a 12-hr light/dark cycle environment in a clean facility and given free access to food pellets and water. Pridopidine is dissolved in saline (vehicle), and administered daily by intraperitoneal (i.p.) injection at a dose of 5 or 6 mg/kg per bodyweight during the light phase of the circadian rhythm. Control mice (WT and R6/2) are injected daily with the same volume of vehicle. All the mice are singly housed in home cage. Pridopidine (5 mg/kg) is administered to pre-symptomatic mice starting at week 5 to week 11 (6 week duration) and for symptomatic animals starting from week 7 to week 9 (3 weeks duration) and 1 week of daily administration (6 mg/kg) at week 10[2].

References:

[1]. Geva M, et al. Pridopidine activates neuroprotective pathways impaired in Huntington Disease. Hum Mol Genet. 2016 Sep 15;25(18):3975-3987.
[2]. Squitieri F, et al. Pridopidine, a dopamine stabilizer, improves motor performance and shows neuroprotective effects in Huntington disease R6/2 mouse J Cell Mol Med. 2015 Nov;19(11):2540-8. model.