Bio Facts About Bio Hacks
Cheat Code or Trojan Horse?
Have you been (bio)HACKED?
SLU-PP-322, Methelyne Blue and other compounds with the aim of improving Mitochondial Health
By: Josh Miller 4/24/2025
Biohacking is a big trend these days, and its certainly not all bad, but sometimes the term can be associated with rushed decisions, contradictions, and sometimes a bit of bro science. It seems like every week some anti aging hack is debunked (if it even had legitimate claims and logic in the first place)…whether it be ice baths or rapamycin or whatever comes next. Again, not all of it is bad, and a lot of the “biohacking” going on is giving options to people who otherwise would be out of luck, its improving quality of life, but in a society where hands go out immediately for a quick solution to a problem pioneers are often guilty of giving less than adequate bandwidth to due diligence and giving the difference to discovery. The result can be that something with therapeutic potential being preemtptively, misapplied or too broadly applied.
Today the topic are two popular research chemicals currently used by people in the hopes of burning fat, increasing endurance, improving mitochondrial health, cognitive health or some combination of the above. SLU-PP-332 and Methylene Blue. Along with an extended discussion of other compounds which are often used with the hope of improving mitochondial health.
This is a bit of a longer read overall, so spoiler alert if you aren’t here for the whole thing; the use of both is a bit more nuanced than some would suggest, and there ARE risks associated with each. Both are cool, and may have some therapeautic potential someday, but depending on a few variables, these compounds could either increase or decrease overall performance, endurance, and even longevity depending on context and application? Are these right for you? If you think “maybe” after reading this and other readings on the subject, find an open minded doctor and bring it up with them. While I hope this helps guide your reading, inform and educate you, and potentially guide you through a conversation around considerations for these compounds with your doctor, please be informed this in no way is intended to diagnose, treat or cure a disease nor is this intended as medical advice. This is for entertainment and educational purposes only. Make sure you know your local laws, and I do not condone or support a violation of any laws in your area. So no using this article to justify acting a fool. Always consult your physician before changing your lifestyle, diet, training, supplement or medicine protocols.
Now then, back to it.
One of the primary topics of discussion when it comes to the benefits of both Methylene Blue and SLU-PP-332 are their benefits for mitochondial health, function and even mitochondrial genesis. Right now MB has a lot of attention, but less people know about SLU-PP-332.
SLU-PP-332 and mitochondrial health
SLU-PP-332 is a small molecule compound that’s a potent and selective stimulator of ERRγ (Estrogen-Related Receptor gamma), a nuclear receptor that plays a significant role in regulating energy metabolism, especially in skeletal muscle, cardiac tissue, and mitochondrial function.
Here’s a breakdown of what’s known
1. Mechanism of Action:
• SLU-PP-332 is an agonistof ERRγ.
• ERRγ is a transcription factor involved in oxidative metabolism, mitochondrial biogenesis, and fatty acid oxidation.
• By activating ERRγ, SLU-PP-332 boosts the expression of genes involved in aerobic metabolism, making tissues more metabolically efficient.
2. Key Effects Observed in Research:
• Increased mitochondrial contentand oxidative capacity, especially in skeletal muscle.
• Improved endurancein animal models (e.g., mice ran longer distances after administration).
• Mimics some aspects of exerciseon a cellular/metabolic level — sometimes called an “exercise mimetic” (though it’s not a substitute for actual exercise).
• May improve insulin sensitivityand reduce fat accumulationin metabolic disease models.
3. Research Context:
• Studied mostly in rodent models.
• Has shown promise in preventing or treating conditions like:
• Type 2 diabetes
• Obesity
• Cardiovascular dysfunction
• Not currently approved for human use — it’s still a research compoundand not available as a supplement or therapeutic.
4. Related Compounds / Pathways:
• Often mentioned alongside PPAR coactivators(like PGC-1α), AMPK, and SIRT1, since these are also involved in energy metabolism.
• ERRγ works downstream or in parallel with these pathways in metabolic adaptation.
A hypothetical case study of the pharmacokinetics of SLU-PP-332 generated by Chat GPT discusscing the application of SLU-PP-332 a female athlete in her 40s who is enhanced with peptides and secretagogues already. She was attracted by the advertising of improved endurance and fat loss.
Caveat: Most available data are preclinical (rodent-based), so any human application is purely speculative and extrapolated from those models. But we can still draw meaningful insights from mechanism and pharmacology.
SLU-PP-332 Overview in Pharmacokinetic Terms
1. Absorption
• Route of administration in studies: Most preclinical studies used intraperitoneal injectionor oral gavagein mice.
• Oral bioavailability in humans: Unknown.However, structural design suggests it was optimized for oral delivery.
• Like many small molecules, lipophilicitymay allow for decent passive diffusion across membranes, aiding oral absorption.
• If oral, you’d expect Tmax in humans to be between 1–3 hoursbased on its behavior in rodents.
2. Distribution
• SLU-PP-332 has high affinity for ERRγ, which is heavily expressed in:
• Skeletal muscle
• Heart
• Liver
• Brown adipose tissue
This suggests it would preferentially concentrate in metabolically active tissues, especially those involved in oxidative metabolism.
• Expected to cross into nuclear receptorswithin cells, not membrane-bound, so entry into the intracellular spaceis required.
3. Metabolism
• Likely metabolized hepatically via cytochrome P450 enzymes, like most small molecule agonists.
• No identified active metabolites as of current publications — effect appears to be from the parent compound.
• Half-life in rodent studies is unknown, but the effects on gene expression were sustained for many hours to days, suggesting a lasting genomic effect even after clearance.
4. Excretion
• Presumed hepatic metabolism and renal or fecal excretion, but specifics aren’t publicly available.
• Notably, because it modulates transcription, its downstream effects can last longer than the drug’s presence in plasma.
Mechanistic Summary Relevant to the Hypothetical Athlete Profile
A. Endurance Enhancement
Mechanism:
• SLU-PP-332 enhances mitochondrial biogenesisand oxidative enzyme expressionvia ERRγ.
• Promotes a shift toward oxidative muscle fiber phenotype(Type I / IIA).
• Increases capillary density and oxygen utilization.
Expected Hypothetical Outcome:
• Improved VO₂ efficiency.
• Delayed fatigue onset during submaximal efforts.
• May allow for greater training volume and faster recoverybetween aerobic sessions.
PK Consideration:
• A once-daily dose (assuming oral bioavailability is optimized) could be enough to stimulate mitochondrial gene expression chronically.
B. Fat Loss and Leaning Out
Mechanism:
• ERRγ activation upregulates genes for:
• Fatty acid oxidation
• Mitochondrial uncoupling
• Glucose sparing(better partitioning)
• Decreases lipid storage signalsin adipose tissue.
Potential Outcome:
• Improved metabolic flexibility— burn fat more efficiently during both rest and exercise.
• Could help maintain or improve insulin sensitivity, which is critical for women in midlife due to perimenopausal shifts in hormone balance.
PK Consideration:
• Sustained receptor activation likely needed over days to weeks to see measurable body composition changes.
C. Strength and Recovery
Indirect Support:
• Not anabolic per se, but improved mitochondrial function may:
• Enhance recovery between sets and sessions
• Support muscle preservationduring fat loss due to more efficient metabolism
• May have cardioprotective effects, especially relevant for women in their 40s+.
And while all this is possible and some are reporting positive anectdotal results, we also do not know the longterm effects, we have no studies in humans, we aren’t sure of the dose in humans, but for mouse models, they used ~20–50 mg/kg/day. So 2-5g per day as a 100kg lifter. 1.5g per day for a lot of female lifters. And it would be presumed to be administered by an oral capsule, but I have mostly heard of this being taken sub Q like many other research chemicals.
Risks and Unknowns
• Long-term effects in humans unknown
• Hormonal effects: ERRγ is part of the nuclear hormone receptor family — there could be unintended crosstalk with estrogen signaling, particularly in perimenopausal women
• Cancer concerns: ERRγ is overexpressed in some tumors — unknown whether activation could affect dormant or latent cancer risk (highly speculative, no direct evidence yet)
• Potential for off-target gene expression, especially with chronic exposure; you only need 3-6 weeks if you do have a risk to benefit ratio that makes sense to use it.
These risks lead us to an obvious question.
Are mitochondrial benefits seen in already healthy individuals?
1. Preclinical Data Says Yes — But With a Ceiling
Most of the animal studies using SLU-PP-332 have been done in otherwise healthy mice, not diseased models. Even in these models:
• Mitochondrial content increased
• Endurance performance improved
• Markers of oxidative metabolism were upregulated
This suggests that even in a baseline healthy state, ERRγ activation pushes the system further— kind of like flipping on the “train harder” genetic switch without more physical effort.
2. But There Is a Point of Diminishing Returns
In highly fit individuals, especially those who already:
• Train endurance consistently
• Have a balanced diet
• Use effective recovery strategies
…the mitochondria are already:
• Dense
• Efficient
• Well-regulated
Artificially pushing further adaptation can theoretically:
• Create energetic inefficiency(e.g., uncoupling or over-oxidation)
• Induce metabolic stress without benefit
• Interfere with natural hormetic processes(the body’s adaptive stress signaling)
So while short-term usemight give a temporary performance edge, long-term usecould:
• Flatten the adaptive curve
• Possibly suppress natural mitochondrial turnover or signaling pathways (like PGC-1α or AMPK)
SLU-PP-332 Likely Effect
Metabolic dysfunction (e.g. insulin resistance, obesity)- Highly beneficial; improves mitochondrial biogenesis and fuel use
Untrained but health- Likely to show meaningful improvements, similar to exercise initiationns
Trained and metabolically healthy- Minor-to-moderate gains; potential for diminishing returns
Elite endurance athlete- Almost certainly redundant or possibly disruptive to fine-tuned adaptations.
4. Risk of Harm in Healthy People?
• Disruption of homeostasis: ERRγ is not just an “on switch” — it balances many processes including circadian rhythm, fuel selection, and possibly estrogen-related functions.
• Oxidative stress: More mitochondria = more ROS if antioxidant systems aren’t equally upregulated.
• Unknown hormonal crosstalk: Especially in women, ERR family activity can influence or respond to estrogen signaling (though ERRγ is considered orphan, meaning no known endogenous ligand).
So what about Methylene Blue? It is wildly popular right now, with many people believing it is totally harmless and without side effects.
Methylene blue (MB), a synthetic compound with a history of medical applications, has garnered attention for its potential effects on mitochondrial function. There is literature looking at this compound in humans, but there is a limited evidence regarding the use of methylene blue for mitochondrial health in otherwise healthy individuals. The following highlights both the potential benefits and concerns.
Potential Benefits of Methylene Blue on Mitochondrial Function
Research indicates that methylene blue can interact with mitochondria to influence their activity. A study published in Free Radical Biology and Medicine demonstrated that methylene blue has a high affinity for mitochondria and, upon photostimulation, can generate reactive oxygen species, leading to mitochondrial dysfunction. Notably, the study also found that methylene blue disrupts mitochondrial energy metabolism even in the absence of light exposure, suggesting potential adverse effects on cellular energy processes (Klosowski et al.).
In the context of diabetic cardiomyopathy, methylene blue has shown substrate-dependent effects on mitochondrial respiration and oxidative stress. A study in The Canadian Journal of Physiology and Pharmacology reported that methylene blue improved oxygen consumption in rat heart mitochondria energized with complex I and II substrates. However, it also elicited a significant increase in hydrogen peroxide production with complex I substrates, indicating a nuanced impact on mitochondrial function (Privistirescu et al.).
Furthermore, methylene blue’s role in neuroprotection has been explored due to its effects on mitochondrial function. A review in Progress in Neurobiology highlighted that methylene blue can reroute electrons in the mitochondrial electron transport chain, enhancing complex IV activity and promoting mitochondrial activity while mitigating oxidative stress. These properties suggest potential therapeutic applications for neurodegenerative disorders (Rojas et al.).
Considerations and Potential Risks
While methylene blue’s effects on mitochondrial function have been documented in various studies, it is essential to consider the context of these findings. Much of the research has been conducted in disease models or under specific experimental conditions, and the applicability of these results to healthy individuals remains uncertain. There is some research to suggest it can cause mitochondial dysfuction and even DNA integrity in healthy people rather than helping. Especially for those in the sun a lot, tanning, or who use red lights… so a lot of the bodybuilding and biohacking community would have some considerations to make for sure. Also, with metal being a potential catylst for these problems and the liklihood of other “supplements” you are using or these supplements themselves being an issue is a concern. (Davies; Yusuke)
Moreover, methylene blue can cause side effects, including dizziness, confusion, and headache. It may also interact adversely with certain medications, such as antidepressants, leading to serious conditions like serotonin syndrome. Therefore, its use should be approached with caution, particularly in self-medication scenarios (Alan).
Conclusion
The current body of research presents a complex picture of methylene blue’s impact on mitochondrial function. While there are indications of potential benefits, especially in disease contexts, the evidence is not conclusive regarding its efficacy and safety for enhancing mitochondrial health in otherwise healthy individuals. Given the potential for adverse effects and the lack of robust clinical data in healthy populations, individuals should exercise caution and consult healthcare professionals before considering methylene blue for mitochondrial health purposes. If you have a legitimate metabolic disorder your physician may determine MB is a good option for you, but just because you feel sluggish so you want to supercharge the “powerhouse of the cell”—not a good enough reason in my opinion. The risk just outweighs the potential reward outside of a disease treatment context supervised by a medical professional.
Why would these drugs cause problems in healthy people but help those with mitochondial dysfunction?
Well, they all turn the dial up on producing more mitochondia without turning up the dial on taking out the trash with mitophagy.
Big Picture
For a recreational athlete, here’s the most honest interpretation:
• If you feel like your metabolism has slowed, or you struggle with recovery, endurance, or leaning out, and you aren’t overtraining, a mitochondrial push mighthelp.
• If you’re already feeling strong, energized, and metabolically stable, there’s less benefit and more risk of dampening natural feedback loops with either of the research chemicals we discussed.
Here’s a Better (and Safer) Lineup for Mitochondrial Support and Leaning Out:
1. PQQ (Pyrroloquinoline Quinone)
• Stimulates mitochondrial biogenesis
• Works synergistically with CoQ10
• May support memory, mood, and endurance
• Dosage: 10–20 mg/day
2. Urolithin A
• Promotes mitophagy(clears out damaged mitochondria)
• Enhances muscle enduranceand cellular rejuvenation
• Has human data showing improved muscle function in middle-aged adults
• Dosage: 500–1,000 mg/day (often from Mitopure® or similar)
3. CoQ10 / Ubiquinol
• Classic antioxidant and ETC (electron transport chain) supporter
• Helps energy productionand may improve VO₂ max
• Vital if taking statins or experiencing fatigue
• Dosage: 100–300 mg/day (Ubiquinol is better absorbed)
4. NMN or NR (Nicotinamide Mononucleotide / Riboside)
• NAD+ precursors, support mitochondrial health, DNA repair, and energy levels
• Useful especially in perimenopause/aging contexts
• Dosage: 250–500 mg/day
5. ALCAR (Acetyl-L-Carnitine)
• Shuttles fatty acids into mitochondria for fuel
• May improve mental clarity, endurance, and fat metabolism
• Dosage: 500–2,000 mg/day
6. Alpha Lipoic Acid (ALA)
• Co-factor in mitochondrial enzyme complexes
• Antioxidant that regenerates other antioxidants
• Dosage: 300–600 mg/day (R-ALA preferred)
Lifestyle Synergy is Key
These compounds work best when paired with:
• Fasted Zone 2 cardio(for mitochondrial stress and fat oxidation)
• Strength training(to maintain muscle during leaning out)
• High-protein diet(to protect lean mass)
• Sleep + stress management(mitochondria hate cortisol)
Mitophagy is a new concept for a lot of people, and not one they typically consider with mitochondial health, and Urolithin A is in the above stack for that purpose. So what is it?
Urolithin A (UA) is a gut-derived metabolite — your body makes it if your microbiome can convert ellagitannins (from things like pomegranates, walnuts, raspberries) into UA.
Spoiler: Most people can’t, or only produce very little.
So researchers started developing direct UA supplements — now sold in clinically studied forms like Mitopure®.
Mechanism of Action: Mitophagy Support
Here’s the key difference:
• PQQ, CoQ10, NMN= support mitochondrial biogenesis(making new mitochondria)
• Urolithin A= enhances mitophagy(removal of damaged or dysfunctional mitochondria)
Think of it like Marie Kondo-ing your mitochondria:
“Does this dysfunctional mitochondrion spark ATP? No? OUT.”
This ensures that:
• The new mitochondria (created from your PQQ/NMN stack) are actually useful
• You’re not cluttered with inefficient organelles stealing resources or leaking ROS
Key Human Research Findings:
1. Muscle Endurance
• Study in middle-aged adults:
500–1,000 mg/day of UA for 4 months
Result: Improved leg enduranceand mitochondrial gene expressionwithout exercise.
2. Cellular Energy Production
• Improves expression of mitochondrial proteins (PGC-1α, TFAM)
• Supports ATP production, especially under stress
3. Longevity and Muscle Aging
• In animal studies, it extended lifespanand healthspan
• Preserved muscle mass and function in aged mice even without exercise
PQQ: Mitochondrial biogenesis
UA: clears out weak/dysfunctional mitochondria, leaving room for new ones
CoQ10: ETC + antioxidant; Helps clean mitochondria perform better once UA removes bad ones
NMN/NR: NAD+ replenishment, gene expression; UA-induced mitophagy depends on sirtuin/NAD+ signaling for cleanup
ALCAR: Fat transport, mental energy; New, efficient mitochondria from UA use transported fats more effectively
ALA: Antioxidant + metabolic regulator; Buffers oxidative stress from enhanced mitochondrial turnover
Combine these supplements with Zone 2 cardioto supercharge mitochondrial turnover (they complement each other)
Proper diet, sleep, and managing stress helps everything. Make sure lifestyle aligns with the more novel efforts.
Urolithin A is like a janitor-meets-mechanic for your cellular engines. It doesn’t just pump up your output — it ensures you’re not dragging along damaged parts that make you less efficient. For someone like you — serious about performance, recovery, metabolic clarity, and fat use — UA rounds out the mitochondrial stack beautifully. It’s not hype; it’s the housekeeping partner your other compounds needed.
In my opinion UA is the missing piece in a lot of mitochondial health protocols. You don’t technically need it in every context, but if you’re purposefully stimulating mitochondrial biogenesis (like you are with PQQ, NMN, etc.), then not having something to manage mitochondrial quality control means you’re essentially making new furniture without throwing out the broken stuff.
Why That Balance Matters:
1. Mitochondrial biogenesis = Quantity
• You’re signaling your cells to build more mitochondria.
• This is great… unless you’re also accumulating dysfunctional ones, which:
• Waste energy
• Leak ROS
• Interfere with signaling
• Reduce overall efficiency
2. Mitophagy = Quality
• Urolithin A activates mitophagy via pathways like
• AMPK
• Sirtuins (especially SIRT1/3)
• PINK1/Parkin pathways
• It flags damaged mitochondria and triggers autophagosomal clearance, so your cell runs smoother and cleaner.
Bottom Line: Mitochondrial Remodeling > Just Expansion
You can’t build an efficient engine without first clearing out the broken parts. That’s how your cells work, too. When you’re stacking compounds to enhance energy, performance, and recovery, it’s not just about making more mitochondria — it’s about making sure the right ones survive and thrive.
Doing both = remodeling for efficiency, longevity, and performance.
This system is especially important for:
• Women in their 40s and beyond(as natural mitophagy declines
• Athletes recovering from overtraining
• Anyone using fasting, exercise, or nootropics to optimize metabolism
You don’t need to biohack recklessly — for a little more effort you can run a protocol with more human research backing it, more time showing safety and ultimately one that gives better returns with lower risk. You can’t pretend to be hardcore and willing to do anything if you choose a potentially worse outcome based on convenience. You can’t talk about sacrifices and risks you take while knowingly sacrificing performance and safety at the same time. Be smart, let your hacking be more akin to using cheat codes than downloading a trojan horse.
Works Cited
Andreux, Paul A., et al. “The mitophagy activator urolithin A is safe and induces a molecular signature of improved mitochondrial and cellular health in humans.” Nature Metabolism, vol. 1, no. 6, 2019, pp. 595–603.
Alan, Jamie K. “The Internet Thinks RFK Jr. Uses Methylene Blue—What to Know About the Mysterious Liquid.” Health.com, 19 Feb. 2025
ChatGPT. “Discussion on SLU-pp-322, Mitochondrial Biogenesis, Mitophagy, and Supplement Synergy Including PQQ, Coenzyme Q10, and Urolithin A.” ChatGPT, OpenAI, 25 Apr. 2025.
Davies, J et al. “Methylene blue but not indigo carmine causes DNA damage to colonocytes in vitro and in vivo at concentrations used in clinical chromoendoscopy.” Gut vol. 56,1 (2007): 155-6. doi:10.1136/gut.2006.107300
Klosowski, Eduardo Makiyama, et al. “The Photodynamic and Direct Actions of Methylene Blue on Mitochondrial Energy Metabolism: A Balance of the Useful and Harmful Effects of This Photosensitizer.” Free Radical Biology and Medicine, vol. 153, June 2020, pp. 34–53.
Privistirescu, Anca I., et al. “Methylene Blue Improves Mitochondrial Respiration and Decreases Oxidative Stress in a Substrate-Dependent Manner in Diabetic Rat Hearts.” The Canadian Journal of Physiology and Pharmacology, vol. 96, no. 10, Oct. 2018, pp. 1012–1016.
Rojas, Julio C., et al. “From Mitochondrial Function to Neuroprotection—An Emerging Role for Methylene Blue.” Progress in Neurobiology, vol. 96, no. 1, Jan. 2012, pp. 32–45.
Quinzii, Catarina M., and Michio Hirano. “Coenzyme Q and mitochondrial disease.” Developmental Disabilities Research Reviews, vol. 16, no. 2, 2010, pp. 183–188.
Stites, Thomas E., et al. “Pyrroloquinoline quinone (PQQ) modulates mitochondrial quantity and function in mice.” Journal of Nutrition, vol. 136, no. 2, 2006, pp. 390–396.
Yusuke Hiraku, Hiroyuki Goto, Masaki Kohno, Shosuke Kawanishi, Mariko Murata, “Metal-mediated oxidative DNA damage induced by methylene blue.” Biochimica et Biophysica Acta (BBA) - General Subjects, Volume 1840, Issue 9, 2014, Pages 2776-2782,ISSN 0304-4165, https://doi.org/10.1016/j.bbagen.2014.04.020 (https://www.sciencedirect.com/science/article/pii/S0304416514001536)