WHAT IS NAD+

Nicotinamide Adenine Dinucleotide (NAD+) is a coenzyme found in all living cells. It is a dinucleotide, meaning it consists of two nucleotides joined through their phosphate groups: one containing adenine and the other containing nicotinamide (a form of Vitamin B, or niacin).

Known as NAD+ in its oxidized state and NADH in its reduced state, it was first described more than a century ago as a molecule in the electron transport chain in the metabolic reduction-oxidation reactions in mitochondria. 

WHAT DOES NAD+ DO?

Nicotinamide adenine dinucleotide (NAD)  is a promising new treatment to support age-related conditions via cell regeneration. NAD is an important metabolic regulator of cellular redox reactions and a cofactor or a co-substrate for key enzymes essential for normal cellular function in different tissues. NAD+ is central to over 400 cellular reactions, serving two critical roles that directly impact cellular resilience and aging.

1. Energy Metabolism (Redox Reactions)

NAD+ acts as a necessary cofactor for dehydrogenases, the enzymes that catalyze redox (oxidation-reduction) reactions in major catabolic pathways:

• Glycolysis and TCA Cycle: NAD+ accepts electrons (a hydride ion) from nutrient breakdown, becoming NADH.

• Mitochondrial Function: NADH then donates these electrons to Complex I of the Electron Transport ChainETC in the mitochondria, driving the production of ATP (cellular energy). The ratio of NAD+/NADH is a primary sensor of the cell's energy status.

2. Cellular Signaling and Repair (Non-Redox Reactions)

NAD+ is consumed as a substrate by regulatory enzymes that link metabolism to longevity:

• Sirtuins (SIRTs): NAD+ dependent deacetylases (such as SIRT1) that regulate transcription, DNA repair, inflammation, and metabolic pathways in response to energy availability. As NAD+ levels drop with age, Sirtuin activity declines, compromising cellular health.

• PARPs (Poly-ADP-Ribose Polymerases): Key enzymes that consume large amounts of NAD+ rapidly to detect and repair DNA damage. Age-related DNA damage increases PARP activity, which drains the overall NAD+ pool.

BENEFITS/ CLINICAL TRIALS

NAD levels decline with age. Evidence from animal studies indicate that interventions that increase NAD levels produce numerous benefits on the overall cardiometabolic health and immune function.

A study reviewing the current literature published in the journal Pharmaceuticals identified 36 human clinical trials, 17 reporting beneficial outcomes, while 12 reported no benefits, and the remaining 7 only evaluated pharmacokinetics, safety, and efficacy.

Interventions which were assessed, include acute kidney injury, Alzheimer’s disease, chronic fatigue syndrome, dementia, hyperphosphatemia, hypertension, obesity, Parkinson’s disease, photoaging of skin, psoriasis, skin cancers, type 1 diabetes mellitus, type 2 diabetes mellitus, and schizophrenia.

Despite the claims that NAD IV therapy may benefit Alzheimer's Disease, Parkinson's, Heart disease, Diabetes, Depression, Chronic Fatigue, and cognitive function, there is little if any evidence to support NAD as a therapeutic treatment. 

• Increases energy and mental clarity
• Supports brain health and cognitive function
• Improves sleep quality and mood balance
• Aids in muscle recovery and performance
• Enhances detoxification and immune function
• Promotes healthy metabolism and fat oxidation
• Supports cardiovascular and mitochondrial health

SIDE EFFECTS

Due to its role as a naturally occurring signaling protein, TB-500 is generally considered low in toxicity in preclinical models. However, its unapproved status means the full risk profile in humans is unknown.

• Systemic Effects:

  • Headache or dizziness (usually mild and transient).

  • Lethargy or tiredness (uncommon).

• Local Reactions:

  • Injection Site Reactions: Pain, swelling, or redness at the subcutaneous injection site.

• Unknown Risks: Because Tb4 promotes cell migration and proliferation, there is a theoretical risk regarding its potential interaction with pre-existing or latent malignancies (cancers), which warrants extreme caution and further research.

Legal and Anti-Doping Concerns

Furthermore, legality is another important aspect that users should consider. The regulatory status of TB500 varies by country, and it may be prohibited in competitive sports by organizations like the World Anti-Doping Agency (WADA). TB500 is listed under S2: Peptide Hormones, Growth Factors, and Related Substances, which makes its use banned for athletes in many sports. Athletes should verify the regulations surrounding TB500 in their respective sports to avoid potential disqualifications or penalties.

IS NAD+ SAFE?

• Short-Term Safety: Numerous small-scale human clinical trials (up to 12-24 weeks) have found NAD+ precursors to be safe and well-tolerated.
• Lack of Long-Term Data: Long-term safety studies (over several years) in humans are currently lacking. The consequences of chronically altering NAD+ levels over a lifetime are not yet fully understood.
• Regulatory Status: The U.S. Food and Drug Administration (FDA) has not approved NAD+ as a drug or treatment, nor have they established standard dosing protocols. Supplements are not regulated with the same rigor as medications, meaning manufacturers are responsible for ensuring safety without pre-market FDA approval.
• Potential Cancer Risk (Precautionary): The relationship between NAD+ and cancer is complex. While adequate NAD+ helps with DNA repair, cancer cells also use NAD+ to fuel their rapid growth. An animal study suggested high doses of an NAD+ precursor might increase the risk of aggressive cancer in mice prone to it. Consequently, health experts strongly advise individuals with active cancer, a history of cancer, or a high cancer risk to avoid NAD+ boosters.

DOSAGE

• Week 1-4: 25mg once to three times a week
• Week 5-8: 50mg once to three times a week
• Week 9+: 100mg once to three times a week

Note: Always consult with a healthcare provider before beginning any supplementation regimen.

RECONSTITUTION

When reconstituting a 500 mg vial of lyophilized NAD+ powder, the most common recommendation from research suppliers and compounding pharmacies is to use 5 mL of bacteriostatic water. Refrigerate immediately after mixing.

WHERE TO BUY NAD+

Researchers should always vet their sources to ensure that a few key factors are present in their test subjects. With the rise in peptide popularity in recent years, many companies have created peptides that undergo little to no testing, quality standards, or regulations. As it is not regulated by the FDA, researchers must do their due diligence and look closely at the company's practices and standards. 

When selecting a supplier for NAD+, focus on transparency and quality assurance, not customer testimonials:

1. Quality Documentation: A reputable supplier must provide:
   •  Certificate of Analysis (COA): This document must be recent (corresponding to the batch/lot number purchased) and demonstrate a minimum purity of >95% via High-Performance Liquid Chromatography (HPLC) testing.
   • Mass Spectrometry (MS) Data: The COA must include mass spectrometry (MS) confirmation to verify the compound’s exact molecular weight, confirming its chemical identity.
   • Contaminant Testing: Look for reports on heavy metals, microbial load, and solvent residues (e.g., residual trifluoroacetic acid, or TFA). The presence of these contaminants can severely compromise research and introduce unknown toxicity.
2. Vendor Verification and Transparency 

   • Specialization: Prioritize vendors who specialize in the manufacturing and distribution of peptides for academic and biotechnology research, rather than general supplement vendors.

   • Manufacturing Origin: Inquire about the source of the raw materials and the manufacturing protocols. Ideal suppliers adhere to strict quality control processes.

   • Handling & Storage: The supplier must provide clear documentation on the proper storage and handling procedures for the peptide to maintain its stability and integrity.

Conclusion on Procurement: Given the high risk of contamination, mislabeling, and legal ambiguity. The use of NAD+ outside of this defined research context poses unacceptable, unquantified risks to human health.

REFERENCES

• Body Techs Rx Wellness + Rehab. “The Ultimate Guide to NAD+ : Benefits, Usage, and Storage.” Body Techs Wellness + Rehab 612-345-5963, bodytechsrx.com/nad%2B-%26-peptide-guide#:~:text=Suggested%20Dosage%20Guidelines:,Solution:%20Use%20a%20sterile%20syringe. Accessed 21 Nov. 2025. 
• Hinchman, Walter. “What Is NAD Infusion: NAD IV Therapy Benefits.” Swolverine, Swolverine, 24 Aug. 2022, swolverine.com/blogs/blog/what-is-nad-infusion-therapy?_pos=1&_sid=a27ff41a0&_ss=r. 
• American Chemical Society, www.acs.org/molecule-of-the-week/archive/n/nicotinamide-adenine-dinucleotide.html. Accessed 21 Nov. 2025.

• Trends in Molecular Medicine. "$\text{NAD}^{+}$ in Aging: Molecular Mechanisms and Translational Implications."

• The Lancet Diabetes & Endocrinology. "$\text{NAD}^{+}$ precursor supplementation in human ageing: clinical evidence and challenges."

• The Journal of Clinical Endocrinology & Metabolism. "Weight Loss Is Associated With Increased $\text{NAD}^{+}$/$\text{SIRT1}$ Expression But Reduced $\text{PARP}$ Activity in White Adipose Tissue."

• American Journal of Physiology-Endocrinology and Metabolism. "Evaluation of safety and effectiveness of $\text{NAD}$ in different clinical conditions: a systematic review."

• Molecular Metabolism. "Hypothalamic $\text{NAD}^{+}$-$\text{Sirtuin}$ Axis: Function and Regulation."

• Clinical Trials.gov. (Ongoing trials on $\text{NMN}$ and $\text{NR}$ in human subjects).