MNTF Discovery
Neurodegenerative diseases and disorders are complex, multifactorial conditions, and that multiple biologic targets must be modulated to affect the disease in question. It is not surprising, then, that the current paradigm of single-target drug development continues to result in clinical trial failures with respect to neurodegenerative conditions. And yet for all this complexity, there is a considerable overlap of common pathways and targets across neurodegenerative diseases. It therefore stands to reason that something that could regulate these common underlying pathways and targets could have dramatic effects across a range of neurodegenerative conditions.
Building on this core hypothesis, Genervon discovered what it believes is the naturally occurring regulator of the development of the human nervous system, called the “motoneuronotrophic factor” (MNTF), which is endogenously expressed at its highest levels at week nine of fetal gestation and remains detectable at very low levels in adult tissue. Extensive preclinical testing over many years suggests that MNTF regulates multiple key CNS-related biological functions—including neuronal differentiation, axonal regeneration, reinnervation, and inflammation and apoptosis—providing both neuroprotective and neuro-regenerative therapeutic effects.
This chart shows the change in MNTF expression in human placenta over the course of gestation in weeks. MNTF expression rose rapidly during the 6th to 9th weeks of gestation in placenta with increasing week-age, peaked at the 9th week, and then declined progressively.
GM6 Development
Development of GM6, an innovative first-in-class pleiotropic peptide with broad CNS potential
GM6 is a 6-amino-acid peptide and an active analog of the endogenous embryonic stage regulator MNTF.
GM6 is able to rapidly transit the human blood-brain barrier given its small size and molecular weight.
GM6 has been shown to be safe and well-tolerated in a Phase 1 study, multiple Phase 2A studies, and compassionate use.
GM6 is non-mutagenic and does not inhibit typical CYP isoforms or interact with tPA.
GM6 is manufactured at low cost through solid-phase synthesis under GMP.
GM6 is stable for 120 months at -20°C and 69 months at 5°C.
GM6 and the Blood-Brain Barrier
GM6 Rapidly Transits the Blood-Brain Barrier
GM6 binds rapidly and remains bound in vivo after 4 hours.
GM6 has a good brain-to-plasma ratio (1.65) in biomimetics measurement.
GM6 rapidly transits the blood-brain barrier enabling neuron survival through developmental-stage pathways: reducing inflammation, decreasing oxidative stress, reducing plaque formation, increasing growth factor, and decreasing apoptosis in the central and peripheral nervous systems.
GM6 Efficacy in Neuroprotection
GM6 Demonstrates Efficacy in Neuroprotection from Toxic CSF in Many CNS Diseases in Preclinical Models of Neurodegeneration
Neurons die when exposed to toxicity from postmortem CNS patient’s Cerebrospinal Fluid (“CSF”)
GM6 demonstrated a statistically significant increase in neuron survival (p <0.0001) in:
ALS (175%)
MS (246%)
AD (191%)
HD (273%)
PD (198%)
Stroke (205%)
GM6 Efficacy in ALS Models
GM6 Increases Neurogenesis and Lowers SOD1 and Inflammation to Ameliorate ALS Disease in ALS SOD1 Animal Model
In an ALS SOD1 mice model, in a dose dependent fashion, GM6 improved behavior, survival rate, strength, and clinical score with statistical significance.
In addition, GM6 significantly increased motor neuron survival, reduced hSOD1 protein level, and reduced inflammation biomarkers.
At 1 mg/kg and 5 mg/kg, reduction of cytokines were: TNFα (-50% and -60%), IL-1β (-65% and -80%), and TGF-β (-50% and -65%). Increased NGF (+60% and +160%)
GM6 may modulate ALS disease through regulating inflammation response and increasing NGF.
GM6 Efficacy in Parkinson’s Disease (PD) Models
GM6 in MPTP and 6-OHDA PD animal models improved functions and motor activities and increased dopamine and neuron protection
Intravenous injections of varying doses of GM6 of 1, 5, 10, or 20 mg/kg in saline showed dose response significant improvements in behavioral testing including spontaneous activity and time-to-rotarod failure with statistical significance.
Quantification of brain monoamines with high pressure liquid chromotography (HPLC) showed GM6 significantly increased Monoamine (dopamine [DA] and metabolite dihydroxyphenyacetic acid [DOPAC] and homocanillic acid [HVA] levels in both models.
The figures below show that GM6 increases dopaminergic neurons in the Substantia Nigra Pars Compacta (SNpc) in 6-PHDA mouse Model for PD by Immunohistochemical staining.
GM6 protects the dopaminergic neurons in SNpc from the detrimental effects of PD induction.
GM6 Efficacy in Alzheimer’s Disease (AD) Models
GM6 Attenuates AD Pathology in APP Mice in Multiple Pathways by Reducing A𝝱 peptide and Inflammation and Modulating Key Biomarkers
Genervon investigated the effect of GM6 in 3-month old APP/ΔPS1-Tg double–transgenic mice model before the development of amyloid plaques at ~3 to 4 months.
GM6 treatment demonstrated a dose-dependent, statistically significant (P < 0.05), and favorable outcome in APP mice after 4 months of treatment with 1 or 5 mg/kg of GM6 compared to placebo, with 5mg/kg results as follows with statistical significance:
Reduced accumulation of Aβ peptide ▼80%
Reduced amyloid load ▼71% Aβ₁₋₄₂, ▼66% Aβ₁₋₄₀
Reduced Inflammation ▼84% TNFα, ▼93% IL-1β, ▼93% TGF-β, ▼microglia 70%, ▼astrocytes 71%
Decreased cathepsin B expression ▼62%
Improved spatial orientation by faster travel: ▼escape latency time and distance traveled 56%
Increased NGF levels in brain ▲ 600%, and
Tempered the memory impairment ▼50%
GM6 modulates various pathways early in the disease process including up-regulating APP catabolism to reduce Aβ deposit and effectively alter the disease process in AD.
GM6 Attenuates AD Pathology in h-Tau Mouse Model by Reducing inflammation, Lowering Tau Hyperphosphorylation, and Attenuating Behavioral Changes
In healthy neurons, tau normally binds to and stabilizes microtubules. Abnormal chemical changes cause tau to hyperphosphorylate and form tangles which block the neurons’ transport system causing tauopathy and clinical manifestations of AD. Tau is a downstream target of GM6 as manifested in the lowering of the hyperphosphorylation of tau.
In vitro study with SK-N-SH cells treated with AGE-BSA or aggregated Aβ₁₋₄₂ showed a dose-dependent increase in tau hyperphosphorylation as determined by Western blot analysis of pT231. GM6 treatment reduced pT231 by >50%.
In a mouse tau model (h-Tau) which showed hyperphosphorylation of h-Tau and behavioral deficits, GM6 demonstrated a reduction of pT231 hyperphosphorylated tau (>75%) and attenuated the behavioral changes with statistical significance (>50%) (n=10 animals/group).
Cytokines were extracted from h-Tau mouse brains and measured by ELISA. Quantitative analysis of cytokine levels by ELISA for the vehicle control group compared to the GM6-treated group showed a statistically significant decrease in TNF-α at 60 vs 10pg/mg protein (-83%, p<0.05), in IL-1β at 180 vs 20 pg/mg protein (-89%, p<0.05), and in IL-6 at 120 vs 20 pg/mg protein (-83%, p<0.05).
DNA microarray study indicated that GM6 represses tau (MAPT↓) and down-regulates mitochondrial genes.
GM6 Efficacy in Stroke Model
GM6 Decreases Infarct and Neurological Deficit in MCAO Stroke Model*
1 hour occlusion of middle cerebral artery (MCAO) of mice, drug given IV at beginning of 24-hour reperfusion
Treatment with GM6 reduced infarct size in a dose-dependent manner with statistical significance.
Neurological scores were as follows: 0, normal motor function; 1, flexion of torso and contralateral forelimb when animal was lifted by the tail; 2, circling to the contralateral side when held by tail on flat surface, but normal posture at rest; 3, leaning to the contralateral side at rest; 4, no spontaneous motor activity.
Treatment with GM6 improved motor function as seen in the statistically significant reduction in neurological deficit scores in a dose-dependent manner.