S-Mitochonic Acid 5. Increases ATP, NAD+ and SIRTUINS

Mitochonic Acid-5 (MA-5): A Novel Mitochondrial and Longevity Molecule

After a long wait, my lab has finally completed the synthesis and isolation of the S-enantiomer of Mitochonic Acid-5 (MA-5) with 99% enantiomeric purity.

The original paper that caught my attention on biorxiv is named:

Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation

I can’t post links right now.

MA-5 is a modified version of the natural plant hormone indole-3-acetic acid.

Across multiple studies, MA-5 has been shown to:

  • Increase ATP production
  • Reduce mitochondrial ROS generation at the source
  • Elevate intracellular NAD⁺ levels
  • Increase and stabilize SIRT1–7 protein expression
  • Improve cellular survival and stress resistance
  • Extend lifespan and healthspan in animal models
  • Protect against hearing loss and improve auditory resilience

I have just tried the R-enantiomer so far and it gives an increase in energy, endurance, mood, and has a very nice effect of making your hearing sharper. The S-enantiomer should have even stronger effects.

To understand how MA-5 actually works, let’s first establish a clear picture of how mitochondria produce energy:

The Engine: Proton Pressure and Turbines Mitochondria generate ATP by creating a high-pressure buildup of protons across the inner mitochondrial membrane and using that pressure to spin a biological turbine called ATP synthase (Complex V).

To fuel this process, the Krebs cycle (or Citric Acid Cycle) first acts as a refinery within the mitochondrial matrix. It processes breakdown products from carbohydrates, fats, and ketones to strip away high-energy electrons, loading them onto specialized carrier molecules (primarily NADH).

These carriers deliver the electrons to four protein complexes (Complexes I–IV) embedded along the folded inner membrane. As electrons pass through these complexes, their energy is used to pump protons from the matrix into the intermembrane space, creating a positive proton gradient.

Finally, ATP synthase uses the mechanical force of these protons flowing back into the matrix to join ADP and inorganic phosphate together. This spins the turbine and produces ATP.

Why Structure is Crucial Efficient energy capture depends entirely on the 3D architecture of the inner membrane. The membrane folds into tight, disc-like pockets called cristae, which act like pressurized batteries.

  • The Pumps (Complexes I–IV): These sit along the flat sides of the cristae pocket, pumping protons inside to inflate it with chemical pressure.
  • The Turbines (ATP Synthase): To capture this energy, ATP synthase molecules cluster into long rows (oligomers) along the sharp, curved ridges of the cristae. This geometry concentrates the trapped protons at the tips, forcing them to exit through the turbines at maximum velocity.

The structural integrity of this system relies on the Crista Junction—the narrow “neck” where the crista pocket connects to the mitochondrial wall. The protein responsible for organizing this junction is Mitofilin (MIC60), a core component of the MICOS complex.

Mitofilin functions like a reinforced ring that lines the junction opening. It circles the narrow neck to prevent it from tearing open or collapsing. Simultaneously, it acts as a structural staple, pinning this ring to the outer mitochondrial membrane.

This architecture maintains a tight, reinforced “bottleneck” at the junction, which is critical for two reasons:

  • It traps protons: By keeping the neck narrow, it prevents protons from leaking out of the pocket, maintaining high pressure for the turbines.
  • It stabilizes the ridges: By anchoring the base, it allows the cristae to extend deep into the matrix without collapsing, enabling ATP synthase to align properly at the tips.

How MA-5 Improves Mitochondrial Structure MA-5 is effective because it strengthens the Mitofilin (MIC60) at the crista junction by binding to it, preventing it from widening or breaking, a common failure in stressed or aging mitochondria that leads to depressurization and structural collapse.

When the Mitofilin is reinforced by MA-5, the downstream benefits are profound:

  • Restored Geometry: The cristae pockets remain inflated and structurally sound.
  • Optimized Alignment: ATP synthase at the distant ridges is forced into its efficient, oligomerized state.
  • Smoother Flow: With the structure intact, the electron transport chain (Complexes I–IV) operates without stalling, significantly reducing the leakage of electrons that causes oxidative stress (ROS).

Rather than just pushing the engine to run harder, MA-5 optimizes and restores the physical conditions that allow energy production to occur efficiently and cleanly.

Mitofilin (Mic60) levels drop significantly with age and disease, and this appears to be a major driver of mitochondrial breakdown across multiple conditions including Parkinson’s. MA-5 may help prevent or delay onset of different mitochondrial-driven diseases.

Part 2: The Signaling Mechanism

Molecular Pharmacology of the S-Enantiomer While the structural stabilization of mitofilin provides the physical scaffold for energy production, the S-enantiomer of MA-5 drives a distinct metabolic and signaling cascade. The study identifies this enantiomer as the pharmacologically active form responsible for upregulating the NAD⁺/Sirtuin axis through a novel “non-DNA-damaging” signaling pathway.

1. Direct Agonism of NAMPT Increases NAD⁺ NAD⁺ levels are critical for mitochondrial function and sirtuin activity but naturally decline with age. The S-enantiomer addresses this by acting as a direct chemical agonist of NAMPT (nicotinamide phosphoribosyltransferase), the rate-limiting enzyme in the NAD⁺ salvage pathway.

  • Binding Mechanism: Structural analysis and docking simulations confirm that the S-enantiomer binds to the active site of NAMPT.
  • Enzymatic Activation: This binding allosterically enhances the enzyme’s activity, resulting in a significant dose-dependent increase in intracellular NAD⁺ levels. Notably, this effect is specific to the S-enantiomer; the racemic mixture and R-enantiomer show significantly weaker or negligible effects on NAD⁺ synthesis.

2. Sirtuin Stabilization: Stopping the “Disposal” System Typically, increasing NAD⁺ just gives the existing sirtuins more fuel. MA-5 is unique because it actually increases the number of sirtuin proteins. It achieves this by blocking the cellular process that normally identifies and destroys them.

  • The “Safe” Repair Signal: MA-5 activates DNA-PK, a master sensor usually linked to DNA repair. Crucially, it triggers this sensor without causing any actual damage to the DNA. It effectively tricks the cell into entering a “protective repair mode” without the stress of an actual injury.
  • Halting Degradation: This signal modifies a regulator protein called TRIM28. Once modified, TRIM28 stops the cell from tagging sirtuins for destruction (a process called ubiquitination).
  • The Result: Instead of being broken down and recycled as usual, sirtuin proteins are preserved and allowed to accumulate. This ensures that the newly boosted NAD⁺ levels have a larger pool of enzymes ready to use that fuel for cellular repair.

The ability of MA-5 to optimize structure for ATP production, raise NAD⁺ levels, and stabilize sirtuins makes it, in my opinion, the most compelling mitochondrial therapeutic currently in human trials.

I will add more study references in future posts or edit this one.

Hi, this looks interesting. I’ll dig into the research on this a little more.

Here is the paper you mentioned:

Non-DNA-damaging DNA-PK activation improving hearing and prolonging life due to NAD+ and SIRT upregulation (BioRxiv)

And here is my Gemini Summary and Analysis of that paper:

Institution: Tohoku University Graduate School of Medicine (Japan) Source: bioRxiv Preprint (DOI: 10.1101/2025.04.18.649305)

This study presents a radical shift in mitochondrial pharmacology. Researchers have isolated the S-enantiomer of the mitochondrial drug Mitochonic Acid 5 (MA-5) and identified a “therapeutic triad” mechanism that extends survival in a fatal mitochondrial disease model. While MA-5 was previously known to boost ATP by binding to the mitochondrial protein mitofilin, this paper reveals that the S-enantiomer specifically binds to the enzyme NAMPT (the rate-limiting enzyme in NAD+ salvage), significantly boosting NAD+ levels.

Crucially—and controversially—the study claims MA-5 activates DNA-PK (DNA-dependent protein kinase) without inducing DNA damage. This activation triggers the phosphorylation of TRIM28, which subsequently stabilizes SIRT1–7proteins, preventing their degradation. This contradicts the prevailing longevity dogma that DNA-PK inhibition is pro-longevity. The authors argue this unique “non-damaging” activation mimics the regenerative signature of iPSC reprogramming (OSKM factors) and rescues hearing loss and lifespan in Leigh Syndrome mice.

Impact Evaluation: The impact score of this journal is 0 (Preprint), evaluated against a typical high-end range of 30–60+ (e.g., Cell, Nature). Therefore, this is a Preliminary finding. While the authors are established (Tohoku University), this work has not yet passed peer review.


Part 2: The Biohacker Analysis

Study Design Specifications

  • Type: In vivo (Mice) and In vitro (Human fibroblasts, HEI-OC1 inner ear cells).
  • Subjects: Ndufs4 KO mice (a standard model for Leigh Syndrome/Mitochondrial Complex I deficiency).
    • Note: These mice suffer fatal encephalomyopathy and typically die within ~55 days.
    • Groups: Vehicle vs. MA-5 (1 mg/kg and 10 mg/kg).
  • Lifespan Analysis:
    • Control Lifespan: ~60 days (Consistent with historic Ndufs4 KO data).
    • Treatment Lifespan: Significant extension observed (p<0.05), with treated mice surviving past the critical 70-day window where controls died.
    • Crucial Caveat: This is Rescue, not Extension. The study demonstrates preventing premature death in a defective model, not extending the maximum lifespan of healthy wild-type mice (which live ~800+ days).

Mechanistic Deep Dive

The paper proposes a “Triad” mechanism unique to the S-enantiomer:

  1. ATP Surge (Metabolic): Binds Mitofilin (Mic60), physically stabilizing mitochondrial cristae to force ATP production even when Complex I is defective.
  2. NAD+ Refueling (Salvage): Acts as an allosteric activator of NAMPT, the rate-limiting enzyme that converts Nicotinamide to NMN. This is a direct “NAD+ booster” mechanism independent of precursors.
  3. The DNA-PK Paradox (Epigenetic): MA-5 activates DNA-PK without DNA damage signals (like γH2AX). This “clean” activation phosphorylates TRIM28 at Ser824.
  • Result: Phosphorylated TRIM28 stops marking SIRT1 for ubiquitin-mediated destruction.
  • Outcome: SIRT1 (and SIRT2-7) protein levels rise dramatically without needing increased mRNA expression.

Novelty

  • Enantiomer Specificity: It differentiates the S-enantiomer (active on NAD+/SIRT) from the R-enantiomer (inactive on NAD+, active on ATP).
  • NAMPT Agonism: Identifying a small molecule that directly binds and activates NAMPT is a “Holy Grail” target for NAD+ restoration, bypassing the need for NMN/NR supplements.
  • SIRT Protein Stabilization: Increasing SIRT levels by stopping their degradation (via TRIM28) is a novel angle, distinct from “activating” them (like Resveratrol/STACs).

Critical Limitations

  • Disease Model Bias: The lifespan data is derived exclusively from Ndufs4 KO mice. These mice die of specific neurodegeneration. There is zero evidence in this paper that MA-5 extends life in healthy animals.
  • The DNA-PK Contradiction: High DNA-PK activity is typically associated with aging, metabolic decline, and senescence. Several biotech companies are developing DNA-PK inhibitors (e.g., for cancer or aging). This paper argues the opposite—that activation is beneficial—which requires extraordinary evidence not fully provided here.
  • Translational Gap: The paper implies the S-enantiomer is superior, but clinical trials in Japan likely use the racemic mixture (50/50).

Part 3: Claims & Verification

Claim 1: MA-5 S-enantiomer binds and activates NAMPT to increase NAD+.

  • Verification: Internal data only. Search confirms MA-5 is known for Mitofilin binding (ATP). The NAMPT binding claim is novel to this specific preprint.
  • Hierarchy: Level D (In vitro protein binding assay / Docking simulation).
  • Assessment: Plausible but Unverified. A small molecule NAMPT activator (SBI-797812 is a known reference) exists, so the mechanism is chemically possible.

Claim 2: MA-5 activates DNA-PK to stabilize SIRT1 via TRIM28.

  • Verification: Search results confirm TRIM28 (KAP1) regulates SIRT1 stability and that DNA-PK can phosphorylate TRIM28. However, DNA-PK activation is usually a stress/damage response.
  • Hierarchy: Level D (In vitro mechanistic pathway analysis).
  • Assessment: Controversial. The distinction between “damage-induced” and “drug-induced” DNA-PK activation needs independent replication.

Claim 3: MA-5 improves hearing and prolongs survival.

  • Verification: Confirmed in Level D (Mouse Model).
  • Translational Uncertainty: High. The mouse model (Ndufs4 KO) represents a rare pediatric mitochondrial disease (Leigh Syndrome), not normal aging.
  • Safety Check: Phase 1 safety trials for MA-5 (racemic) were completed in Japan (jRCT2031210495) for mitochondrial disease. No “Phase 1” results are published in peer-reviewed English journals yet, but progression to Phase 2 suggests no acute lethality.

Claim 4: “Mimics iPSC (OSKM) reprogramming signatures.”

  • Verification: Based on RNA-seq data within the paper.
  • Hierarchy: Level D (Bioinformatics correlation).
  • Assessment: Speculative. Transcriptomic overlap does not guarantee functional cellular rejuvenation in vivo.

Part 4: Actionable Intelligence

The Translational Protocol

  • Compound: Mitochonic Acid 5 (MA-5).
    • Note: Currently an investigational drug in Japan. Not available as a supplement.
  • Human Equivalent Dose (HED):
    • Mouse Dose: 10 mg/kg (High efficacy dose).
    • Calculation: 10 mg/kg×(3/37)≈0.81 mg/kg.
    • For 70kg Human: ≈57 mg per day.
    • Comment: This is a notably low dose for a small molecule, suggesting high potency.
  • Pharmacokinetics:
    • Oral bioavailability is confirmed (mice fed chow).
    • Target: Mitochondria (Inner Membrane).
    • Half-life: Unknown in humans.
  • Safety & Toxicity:
    • Phase 1 Status: Completed in Japan (healthy adults).
    • Toxicology: No acute toxicity reported in mice at therapeutic doses.
    • Target Organs: Kidney and Heart (primary sites of mitochondrial density). Monitor Cystatin C and Troponin as safety precautions.

Biomarker Verification Panel

  • Primary Efficacy:
    • NAD+ Levels (PBMC): Should rise if NAMPT activation works.
    • ATP Production (Fibroblasts): Can be measured in patient-derived cells.
  • Downstream Targets:
    • SIRT1 Protein Levels: Western blot of PBMCs (not just mRNA, as the mechanism is protein stabilization).
    • Audiometry: High-frequency hearing preservation (8–16 kHz).

Feasibility & ROI

  • Availability: None. It is a proprietary clinical asset (Tohoku University / Miyarisan Pharmaceutical).
  • Sourcing Warning: “MA-5” sold by chemical vendors is likely Racemic. The study claims the S-enantiomer is 2x more effective for NAD+, but Racemic still contains 50% S-enantiomer.
  • Cost-Benefit: If accessible, 50mg/day is highly cost-effective compared to grams of NMN.

Part 5: The Strategic FAQ

Q1: Does MA-5 extend lifespan in healthy adults, or just those with mitochondrial disease? Answer: Unknown. The study only utilized Ndufs4 KO mice (Leigh Syndrome). There is no data provided for wild-type lifespan extension. It acts as a “rescue” agent for defective mitochondria.

Q2: How does MA-5 compare to taking NMN or NR? Answer: Theoretically superior. NMN/NR are precursors that feed into the salvage pathway. MA-5 (S-enantiomer) activates NAMPT, the engine of that pathway. If the engine is sluggish (common in aging), adding fuel (NMN) helps less than fixing the engine (MA-5).

Q3: I am taking Rapamycin. Will MA-5 interfere? Answer: Potential Conflict. Rapamycin inhibits mTOR to induce autophagy and often lowers protein synthesis/ATP initially to signal stress. MA-5 forces ATP production and stabilizes proteins (SIRT1). They might pull metabolic signaling in opposite directions (Survival mode vs. Growth/Rescue mode).

Q4: I am taking Metformin. Is this a bad combo? Answer: Likely Antagonistic. Metformin works by inhibitingComplex I to raise AMP/ATP ratios and trigger AMPK. MA-5 bypasses Complex I defects to restore ATP. MA-5 could theoretically neutralize the hormetic stress signal that makes Metformin effective.

Q5: Isn’t activating DNA-PK bad for aging? Answer: Yes, generally. Chronic DNA-PK activation is linked to senescence and metabolic decline. This paper argues MA-5 activates it “without damage,” but this is a dangerous nuance. If the paper is wrong, MA-5 could accelerate senescence.

Q6: Can I buy MA-5? Answer: No. It is currently in Phase 2 clinical trials in Japan for mitochondrial diseases. Chemical suppliers sell it for in vitro research only, often not pharmaceutical grade.

Q7: Is the S-enantiomer stable, or does it convert to R in the body? Answer: Stable. The study confirmed in Rhesus monkeys that no chiral interconversion occurs in vivo. You must take the S-form (or racemic) to get the NAD+ benefit.

Q8: What specific blood test would show this is working? Answer: intracellular NAD+ (not serum) and SIRT1 protein levels in leukocytes. Standard panels (CRP, metabolic) won’t catch this specific mechanism.

Q9: Does this help with tinnitus or just hearing loss? Answer: The study specifically tested hearing thresholds (ABR). It rescued hair cell death. If tinnitus is caused by cochlear hair cell damage (sensorineural), it might help, but the data is for threshold recovery, not tinnitus suppression.

Q10: What is the biggest risk with this compound? Answer: Tumorigenesis. MA-5 activates survival pathways (SIRT1, ATP) and DNA repair (DNA-PK) in cells that should perhaps undergo apoptosis (like cancer cells). By forcing cell survival and ATP generation, it could theoretically protect early-stage tumors, similar to the concern with antioxidants.

And a deeper analysis and report using Google Gemini Pro Deep Search:

Critical Report: The S-Enantiomer of Mitochonic Acid 5 (MA-5) — A Structural and Metabolic Modulator of Mitochondrial Function

Executive Summary: A Bayesian Evaluation of Therapeutic Potential

This comprehensive report evaluates the scientific standing, mechanistic plausibility, and translational potential of Mitochonic Acid 5 (MA-5), with a specific and critical focus on its S-enantiomer. As the longevity and biotech sectors increasingly scrutinize “mitochondria-homing” therapeutics, distinguishing between hype-driven supplements and mechanism-backed pharmaceutical candidates is paramount. MA-5 represents a divergence from classical antioxidant strategies (e.g., MitoQ, SkQ1) which primarily scavenge reactive oxygen species (ROS). Instead, MA-5 operates as a structural bio-energetic corrector.

Our analysis, based on a rigorous review of pre-clinical and early-phase clinical data, suggests that MA-5 functions through a dual-mechanism “therapeutic triad” that is stereospecific. While the racemic mixture has shown efficacy in stabilizing mitochondrial architecture, the S-enantiomer specifically confers a gain-of-function by activating the NAD+ salvage pathway via NAMPT.

Key Findings & Confidence Assessments:

  • Mechanism of Action (Structure): [Confidence: High]. The binding of MA-5 to Mitofilin (Mic60) and the subsequent oligomerization of ATP synthase is well-supported by biochemical assays and crystallography.1 This mechanism explains the drug’s ability to maintain ATP production even in the presence of electron transport chain (ETC) inhibitors.
  • Mechanism of Action (Metabolism - S-Enantiomer): [Confidence: Medium-High]. Recent bioanalytical data identifies the S-enantiomer as a ligand for NAMPT, initiating a DNA-PK > TRIM28 > SIRT1 signaling cascade.3 This represents a novel pathway for intracellular NAD+ elevation distinct from precursor supplementation.
  • Efficacy in Genetic Disease: [Confidence: High]. The drug demonstrates near-universal rescue (96%) in fibroblast lines from patients with diverse mitochondrial DNA (mtDNA) mutations.1
  • Lifespan Extension (Wild-Type): [Confidence: Low / Null]. Contrary to some enthusiasm in the biohacking community, current data does not support lifespan extension in healthy, wild-type organisms. C. elegans studies show healthspan benefits (motility, neuroprotection) but no extension of median lifespan in wild-type strains.5
  • Safety Profile: [Confidence: Medium]. Phase 1 trials in healthy humans exhibited no serious adverse events.6 However, the theoretical risk of oncogenesis via NAMPT over-activation remains a critical long-term safety concern that requires surveillance.

The following report details the molecular architecture, pharmacological mechanisms, pre-clinical efficacy data, and the specific translational gaps that currently exist between murine models and human longevity applications.

Full report here: https://gemini.google.com/share/689199715486

and another deep search analysis:

The Pharmacological and Geroprotective Profile of the S-Enantiomer of Mitochonic Acid 5 (MA-5): A Critical Evaluation of Mitochondrial Modulation, NAD+ Flux, and Healthspan Extension

1. Introduction: The Bioenergetic Imperative in Aging and Disease

The preservation of mitochondrial function is increasingly recognized as the linchpin of metabolic health, longevity, and resistance to degenerative disease. Mitochondria, often reduced to the simplistic moniker of “cellular powerhouses,” are in reality complex signaling hubs that dictate cell fate, regulate calcium homeostasis, and govern the rate of biological aging. Dysfunctional mitochondria are the etiological root of a spectrum of pathologies ranging from rare, orphan genetic disorders like MELAS (Mitochondrial Encephalomyopathy, Lactic Acidosis, and Stroke-like episodes) and Barth syndrome to pervasive age-related conditions such as sarcopenia, chronic kidney disease (CKD), and sensorineural hearing loss.

Historically, therapeutic interventions for mitochondrial dysfunction have been largely palliative or limited to “cocktails” of antioxidants and cofactors—such as Coenzyme Q10, idebenone, and various vitamins—which attempt to scavenge reactive oxygen species (ROS) or bypass defective electron transport chain (ETC) complexes. However, these strategies often fail to address the structural disintegration of the mitochondrial network or the collapse of the ATP-generating machinery itself. They ameliorate symptoms without rectifying the underlying bioenergetic deficit.

Mitochonic Acid 5 (MA-5), a synthetic derivative of the plant growth hormone indole-3-acetic acid, represents a paradigm shift in this therapeutic landscape. Unlike its predecessors, MA-5 does not merely act as an electron donor or antioxidant. Instead, it functions as a “mitochondria-homing” drug that physically stabilizes the mitochondrial inner membrane organizing system (MINOS) and allosterically modulates key metabolic enzymes. While early research characterized MA-5 as a racemic mixture, recent advancements in stereochemical bioanalysis have stratified the pharmacological activities of its enantiomers. The emerging consensus identifies the S-enantiomer as the eutomer—the biologically active form responsible for the profound metabolic reprogramming capabilities of the drug, particularly concerning the NAMPT/NAD+/Sirtuin axis.

This report provides an exhaustive evaluation of the scientific and clinical research surrounding the S-enantiomer of MA-5. It synthesizes data from structural biology, invertebrate aging models, mammalian toxicology, and early-phase human clinical trials to construct a holistic view of S-MA-5’s potential to not only treat mitochondrial disease but to extend healthspan and compress morbidity in the aging population.

Full Report: https://gemini.google.com/share/0c12c5db7ad0