Proof-of-Concept Studies in Animals

Proof of Concept

Numerous proof-of-concept studies have been performed in several European countries (UK, France, Germany, Finland, Netherlands, Portugal, and Russia) by world-class scientists in top research centres including Maastricht University, C. Bernard University, University of Wuerzburg, Universidade Nova de Lisboa, and Oxford University.

Proof-of-Concept Studies in Animals

The following results have been obtained in proof-of-concept studies in animals:

Mitocholine significantly ameliorates cognitive deficits of different origin:

cognitive deficits caused by normal aging in mice; battery of tests included open field, step-down avoidance, Morris water maze, and 1H NMR in vivo (Storozheva et al, 2008);
cognitive deficits caused by chronic cerebral hypoperfusion in two-vessel occlusion model of vascular dementia and AD in rats; battery of tests included step-through passive avoidance, Morris water maze, and 1H NMR in vivo tests (Storozheva et al, 2008; Pomytkin et al, 2007);
cognitive deficits induced by the administration of beta-amyloid peptide 25-35 into the nucleus basalis magnocellularis in model of amyloid-induced AD in rats; battery of tests included step-through passive avoidance and cortex ChAT activity tests (Storozheva et al, 2008);
cognitive deficits induced by acute scopolamine, a muscarinic antagonist, in scopolamine-induced amnesia model in rats; passive avoidance test (Report C126007, 2007);
cognitive deficits induced by chronic stress-induced disruption of contextual learning and memory in chronic stress model in mice; battery of tests included step-down avoidance and fear conditioning tests (Cline et al, 2012 and 2015);
hyperactivity in TG2576 Alzheimer’s mice; battery of tests included open field and Y-maze (Report C126107, 2007);

Mitocholine significantly increases capacity of cholinergic system in the brain:

increases activity of Choline Acetyltransferase (ChAT), a key enzyme of acetylcholine synthesis, diminished by the injection of beta-amyloid peptide 25-35 into the nucleus basalis magnocellularis; ChAT assay (Storozheva et al, 2008);
prevents disruption of learning performance induced by scopolamine, a muscarinic antagonist; passive avoidance test (Report C126007, 2007).

Mitocholine significantly protects the brain against cerebral hypoxia:

it significantly slowed dynamics of whole-brain decline of energetic substrates ATP and phosphocreatine pCr in the course of global ischemia in cardiac arrest model in rats; 31P NMR in vivo (Pomytkin et al, 2005);
it significantly protected brain mitochondria from hypoxia-induced swelling in micro-stroke model in Thy1-CFP mice; two-photon laser fluorescent microscopy in vivo (Report 2012, Neurotar);
it significantly reduced size of endothelin-1-induced lesions in the brain in stroke model in rats (report 2014, Dr. Daniel Anthony);
it significantly increased whole-brain N-acetylaspartate diminished by chronic cerebral hypoperfusion in two-vessel occlusion model of vascular dementia and AD; 1H NMR in vivo (Storozheva et al, 2008);
it significantly ameliorated cognitive deficits caused by chronic cerebral hypoperfusion in two-vessel occlusion model of vascular dementia and AD in rats; step-through passive avoidance test (Storozheva et al, 2008; Pomytkin et al, 2007).

Mitocholine exhibits significant antidepressant-like effects:

in young adult mice; forced swim test (Cline et al, 2015);
in aged mice; sucrose preference test (Cline et al, 2015);
in young adult mice in model of chronic stress-induced depression; forced swim test and sucrose preference test (Cline et al, 2012);
in young adult mice in model of high cholesterol diet induced depression; forced swim test (Strekalova et al, 2015a);
in young adult mice in the course of chronic delivery of Mitocholine via food pellets; tail suspension test, forced swim test, and sucrose preference test (Costa-Nunes et al, 2015).

Mitocholine exhibits significant anxiolytic-like effects:

in young adult mice in model of chronic stress; dark/light box test (Cline et al, 2012);
in young naïve mice; O-maze test (Cline et al, 2015).

Mitocholine affects neuroplasticity in the insulin-like fashion:

in young adult mice, it significantly upregulated hippocampal expression of set of genes, 41% of which are involved in synaptic plasticity and some of them (Arc, SGK1, and VGF) are known to be upregulated by insulin; Illumina assay (Cline et al, 2015);
in young adult mice, it significantly inhibited GSK-3β, an enzyme playing the role in several types of synaptic plasticity opposed to those for insulin, via phosphorylation on Ser9; GSK-3β phosphorylation assay (Cline et al, 2015);
in young adult mice in chronic stress model, it significantly increased hippocampal expression of IGF2, an agonist of neuronal isoform A of insulin receptors and activator of hippocampal neurogenesis; real-time PCR assay (Cline et al, 2012);
in young adult mice in 5-days stress model, it significantly increased neurogenesis in hippocampal dentate gyrus;
DCX+ and Ki67 cell assay (Report, 2015. Dr. Strekalova);
in young adult mice in chronic stress model, it significantly prevented stress-induced increase in NR2A/NR2B subunit ratio in NMDA receptors, i.e. maintains enhanced plasticity; real-time PCR assay (Cline et al, 2015);
in TG2576 Alzheimer’s mice, it significantly reduced pathologically high activity of insulin degrading enzyme, i.e. prevented excessive degradation of insulin in brains of these mice (Report C126107, 2007).

Mitocholine upregulates mitochondrial biogenesis in the brain

via upregulation of expression PPARGC1b, a master transcription co-regulator (Strekalova et al, 2015).

Mitocholine exhibits significant antiepileptic effects:

ameliorated epilepsy-like seizures and increases life span; corazole-induced model of primary generalized epilepsy in rats (Rychikhin et al, 2013).

Mitocholine exhibits significant effects in Parkinson’s disease model

ameliorated Parkinson’s disease-like motor deficits and muscle rigidity; MPTP-induced model of parkinsonism in rats (Sariev et al, 2011).