Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: Changes in A) NRF2 and B) HSF1 targets 24 hours post dose after 7 days of dosing subcutaneously with different concentrations of M102 (N=3 per group). C) Diagram showing the design of the TDP-43 Q331K study with the major timepoints for electrophysiology and gait analysis. D) Body weight over time showing a reduction in the 2.5mg/kg BD M102 group when compared to vehicle dosed (N=13-14 per group). E) Area under the curve analysis of body weight showing reduction in both M102 dosed groups when compared to the vehicle dose group (N=13-14). F) Rotarod data shown as latency to fall (N=13-14 per group). Change in time spent on G) Diagonal paws and H) 3 paws showing significant improvement of the 2.5mg/kg BD M102 group when compared to the vehicle group (N=8 per group). All data shown as mean +/- SD. (* = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001).

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques:

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Compound muscle action potential (CMAP) amplitude of the hindlimb muscle at 6 weeks and 6 months of age and B) Percentage change in CMAP amplitude between 6 weeks and 6 months of age in M102 dosed groups compared to vehicle (N=12-14 per group). C) Change in repetitive stimulation amplitude between the 1st and 10th stimulus (N=12-14 per group). D) Motor neuron counts from lumbar spinal cord in size bins (N=5-6 per group). E) Cerebral cortex NRF2 and HSF1 targets after 3 months of dosing (N=6 per group). All data shown as mean +/- SD. (* = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001).

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques:

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Pharmacokinetic study of M102 following single subcutaneous (sc) dose at 5mg /kg or B) oral dosing for 1-4 days at 25 mg/kg/day PO (QD). Data show plasma concentrations as determined by HPLC-MS, which was equivalent to 5mg/kg/day SC, assuming 20% relative bioavailability In the multiple dose study. Day 4 showed consistent PK profile to Day 1 Pharmacokinetic profile of M102 dosed at 5mg/kg SC and 10 mg/kg PO. Multiple Oral Dose Study Showed Consistent Exposure.

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques:

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Design for the SOD1 G93A oral M102 study showing the main timepoints for electrophysiological and gait analysis. B) Body weights over time (N=8 per group). C) Area under the curve analysis of body weights (N=8 per group). D) Motor neuron counts per ventral horn (N=37-72 ventral horns per group). Compound muscle action potential (CMAP) amplitudes at E) 60 and F) 90 days of age (N= 7-8 per group). G) Percentage change in CMAP amplitude between 60 and 90 days of age (N=7-8 per group). All data shown as mean +/- SEM (* = p<0.05, ** = p<0.01, *** = p<0.001, **** = p<0.0001).

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques:

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Representative western blots showing increased expression of NQO1 upon 10 μM M102 treatment for 48h in sporadic, C9 and SOD1 ALS patient-derived iAstrocytes. DMSO and M102 treated samples of each line are from the same blot but were cropped because there were different conditions in between these lanes. Original full blots are available upon request. B) Quantification shows that iAstrocytes derived from ALS cases with different genotypes display lower baseline levels of NQO1 compared to healthy controls (N=3 technical repeats per cell line; one-way ANOVA followed by Dunnett’s multiple comparisons test), and C) These levels are increased upon 10 μM M102 treatment for 48h (N=3 technical repeats per cell line; two-way ANOVA followed by Šídák’s multiple comparisons test). (D) Representative staining of NRF2 before and after 10 μM M102 exposure in ALS iAstrocytes, and respective quantifications under (E) Baseline levels and in response to M102 treatment for 30min to 24h (N=4-5 technical repeats per cell line). Two-way ANOVA treatment effect: p-value=0.0067. (F) Representative staining of HSF1 before and after 10 μM M102 treatment in sALS and C9-ALS iAstrocytes, and respective quantifications under (G) Baseline levels and upon M102 treatment for 30min - 24h (N=4-5 technical repeats per cell line). Two-way ANOVA treatment effect: p-value=0.0003. Significance: * <0.05; ** <0.01; *** <0.001.

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques: Western Blot, Expressing, Derivative Assay, Staining

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Representative images of 8-OHdG staining and (B) Respective quantifications show reduced levels of oxidized RNA upon 10 μM M102 exposure for 48h in iAstrocytes derived from SOD1, sporadic and C9 ALS patients. N=2-3 per genotype. Two-way ANOVA followed by Šídák’s multiple comparisons test. C) Representative images of misfolded SOD1 before (top panel) and after (bottom panel) exposure to M102 and D) Respective quantifications show reduction of misfolded SOD1 in iAstrocytes derived from SOD1, sporadic and C9 ALS patients upon M102 exposure (N=3 SOD1 ALS, N=3 C9-ALS, N=2 sALS - each with 3 technical repeats). Two-way ANOVA followed by Šídák’s multiple comparisons test. E) Representative immunoblotting images and F) Respective quantifications show a time-dependent reduction of TDP-43 proteinopathy upon M102 treatment (10 μM for 48h) in sALS (n=10, 2-4 technical repeats each) and C9-ALS (n=3, 3-4 technical repeats each; paired t-test) patient-derived iAstrocytes, characterised by a reduction of the fragmented 35kDa band. Significance: * <0.05; ** <0.01; *** <0.001.

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques: Staining, Derivative Assay, Western Blot

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: A) Schematic figure of the MN-iAstrocyte co-culture system previously described ( 70 ). B) EC50 calculated from dose-response curve tested in sALS patient-derived iAstrocyte/MN co-cultures (average of 4 cell lines, 6-8 technical repeats each). C) ALS-patient derived iAstrocytes are toxic to co-cultured MNs under basal conditions, leading to increased MN death compared to healthy controls, and motor neuron survival is rescued upon 10 μM M102 treatment for 48h in sporadic, C9 and SOD1 ALS cases. N=3. (Two-way ANOVA followed by Šídák’s multiple comparisons test). D) RNA sequencing from iAstrocytes treated with 10 μM M102 or DMSO for 48h reveal transcriptomic changes in response to M102 treatment in ALS iAstrocytes. E) Enriched pathways in response to M102 exposure identified in common between C9, SOD1 and sALS iAstrocytes show that M102 treatment targets various pathophysiological mechanisms known to play a role in ALS. F) Heatmap shows the transcriptomic profile of high and low responders (i.e. >50% rescue and <50% rescue, respectively) to M102 under baseline conditions and upon treatment with 10 μM M102 for 48h. Significance: * <0.05; ** <0.01; *** <0.001.

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques: Co-Culture Assay, Derivative Assay, Cell Culture, RNA Sequencing Assay

Journal: bioRxiv

Article Title: M102, a combined NRF2 and HSF-1 activator for neuroprotection in amyotrophic lateral sclerosis

doi: 10.1101/2024.12.08.627389

Figure Lengend Snippet: These transcriptomic changes are observed across A) SOD1, B) C9orf72, and C) sporadic ALS cases, suggesting that M102 has the potential to be beneficial for both familial and sporadic ALS patients. D) Summary of the DEGs identified between treated and untreated iAstrocytes from healthy controls, C9-ALS, SOD1-ALS and sALS cases.

Article Snippet: GLP general toxicology studies in rats and NHPs were conducted at WuXi AppTec (Suzhou, China) to evaluate potential toxicity of M102 when administered once daily for 28 days by oral gavage and to assess the reversibility, persistence, or delayed occurrence of toxic effects following a 14-day recovery period.

Techniques: