Effects of Trigonelline Monohydrate on NAD+ Metabolism, Pharmacokinetics, Safety, and Tolerability in Healthy Adults

A Randomized, Double-blind, Parallel-arm, Comparator- and Placebo-controlled Clinical Study to Evaluate the Pharmacokinetic and Pharmacodynamic Effects, Bioavailability, Safety, and Tolerability of Trigonelline Monohydrate in Healthy Adults Aged 40 to 70

Nicotinamide adenine dinucleotide (NAD+) is a vitamin B3-derived coenzyme that plays a central role in energy production, cellular metabolism, mitochondrial function, and other physiological processes. NAD+ levels decline with advancing age and this decline has been associated with metabolic dysfunction and age-related physiological changes. Trigonelline is a naturally occurring alkaloid and methylated derivative of nicotinic acid that has been identified as a potential NAD+ precursor. Preclinical and translational research suggests that trigonelline may increase NAD+ levels through the Preiss-Handler pathway. It has also been reported that trigonelline may influence mitochondrial function, muscle health, inflammation, and other metabolic processes. Human clinical data on trigonelline supplementation, however, remain limited.

This Phase 1 study will evaluate the pharmacodynamic effects, pharmacokinetics, bioavailability, safety, and tolerability of trigonelline monohydrate supplementation in generally healthy adults aged 40 to 70 years. The study will use a randomized, double-blind, parallel-arm, placebo-controlled design with an active comparator arm. A total of 60 participants will be randomized in a 1:1:1:1:1:1 ratio to receive one of four daily doses of trigonelline monohydrate (equivalent to 150 mg, 300 mg, 450 mg, or 600 mg of trigonelline daily), 600 mg nicotinamide mononucleotide (NMN; active comparator), or placebo for approximately 4 weeks.

The primary objective is to assess the effect of daily trigonelline monohydrate supplementation on NAD metabolism by evaluating the change from baseline to Week 4 in circulating whole blood NAD+ concentrations measured by liquid chromatography-mass spectrometry (LC-MS).

Secondary objectives include characterizing the bioavailability and pharmacokinetic profile of trigonelline after a single dose, assessing plasma accumulation following repeated dosing, evaluating changes in whole blood NAD-related metabolites over time, and quantifying 24-hour urinary excretion of trigonelline and NAD-related metabolites after acute and repeated dosing. Pharmacokinetic parameters include area under the plasma concentration-time curve (AUC0-24 and AUCinf), maximum plasma concentration (Cmax), time to maximum concentration (Tmax), elimination half-life (T½), and the AUC0-24:AUCinf ratio.

Exploratory objectives include assessment of plasma and peripheral blood mononuclear cell (PBMC) NAD-related metabolites, circulating inflammatory biomarkers (high-sensitivity C-reactive protein \[hs-CRP\], interleukin-6 \[IL-6\], tumor necrosis factor alpha \[TNF-α\], and interleukin-1 beta \[IL-1β\]), and dose-response relationships of trigonelline supplementation on NAD metabolism. Additional samples may be analyzed for gut microbiome composition, metabolomic profiles, short-chain fatty acids, microbial functional pathways, and methylation, oxidative stress, and redox-related metabolites.

Participants will attend a screening visit, a baseline/randomization visit (Day 1), a 24-hour pharmacokinetic and safety visit (Day 2), a Week 1 safety visit, and an end-of-study visit at Week 4. Safety and tolerability will be evaluated through monitoring of adverse events, vital signs, anthropometric measurements, and clinical laboratory assessments throughout the study.

Inclusion Criteria:

* Adults who are 40 to 70 years of age (inclusive) at the time of signing the informed consent.
* Are able to consume study product capsules whole without chewing.
* Have suitable veins for repeated venipuncture.
* Non-smokers (including nicotine vaping) and have not used any nicotine products (patches, gums, etc.) for \> 3 months prior to Visit 2. Non-smoker is defined as someone who does not habitually/regularly use products containing nicotine and willing to abstain for 14 days before Visit 2 until the last procedure.
* In otherwise good general health, as deemed by the investigator (based on a review of medical history, vital signs, laboratory safety tests, and physical examination if performed at screening).
* Have a BMI between 18.5 to 29.9 kg/m2 (inclusive) and weigh no less than 50.0 kg.
* Have maintained consistent dietary habits, including medication and supplement intake, and lifestyle for the last 3 months before screening and agree to maintain them throughout the study (unless required per the restrictions).
* Agree to follow the restrictions on concomitant treatments.
* Agree to follow the restrictions on lifestyle.
* Agree to use acceptable contraceptive methods.
* Willing and able to agree to the requirements of this study, be willing to give voluntary consent, and carry out all study-related procedures.
* Willing and able to comply with the protocol, including:

1. Agree to consume the standardized meals provided at the site during Visit 2
2. Agree to fast overnight, i.e., no food or liquids except for water for a minimum 8 hours prior to applicable visits

Exclusion Criteria:

* Individuals of CBP who are pregnant, intending to breastfeed, or intending to conceive at the time of consent and during the study, or demonstrate a positive pregnancy test.
* Have a known sensitivity, intolerability, or allergy to any of the standardized meals/snacks, study products, their excipients, or to any known allergens that may be present due to potential cross-contamination during manufacturing or processing.
* Have positive screening tests for drugs or cotinine screening.
* Have positive alcohol breath test at screening.
* Have high BP (≥140 systolic or ≥90 diastolic mmHg).
* Have clinically significant abnormal vital signs that in the opinion of the investigator may be of a concern for the study or participant safety.
* Have clinically significant abnormal safety laboratory assessment values in the opinion of the investigator at screening.
* Have current symptoms, or symptoms within the past 7 days of baseline, of an acute infection (e.g., fever, respiratory symptoms such as persistent cough or difficulty breathing), that may influence study outcomes as assessed by the investigator.
* Have Type I or II diabetes, thyroid disease, or non-stable asthma or chronic obstructive pulmonary disease (COPD).
* Reports a clinically significant illness during the 28 days before the first dose of study product.
* Have any abnormality, obstruction, or disorder of the gastrointestinal tract that may preclude swallowing (e.g., dysphagia) or impair gastrointestinal motility, digestion, or absorption (e.g., known intestinal malabsorption, celiac disease, inflammatory bowel disease \[including Crohn's disease or ulcerative colitis\], chronic pancreatitis, or steatorrhea).
* Have any history of gastrointestinal surgery or anatomical abnormality that, in the opinion of the investigator, may affect the oral absorption of the study product.
* Have current or a history of hematological disorders (e.g., sickle cell anemia or thalassemia), cerebrovascular disease, heart/cardiovascular disease (e.g., coronary artery disease, heart failure, stroke, peripheral vascular disease), renal disease (dialysis or renal failure), hepatic impairment/disease, neurological disorders (e.g., dementia or Parkinson's disease), immune disorders and/or immunocompromised (i.e., HIV/AIDS).
* Have a history of cancer (except localized skin cancer without metastases or in situ cervical cancer), unless recovery occurred more than 5 years before the screening visit.
* Are receiving treatments for or have been hospitalized in the last 12 months for psychiatric disorders (e.g., depression, bipolar disorder, schizophrenia, etc.) that may affect the participant's capability to carry out all procedures of the study at the investigator's discretion.
* Major surgery in 3 months prior to screening or planned major surgery during the study.
* Currently consumes, on average, more than 2 standard alcoholic beverages a day where one standard drink is defined as approximately as 330 mL (12 oz) of beer (\~5% alcohol), 150 mL (5 oz) of wine (\~12% alcohol), 40 mL (1.5 oz) of distilled spirits (e.g., vodka, whiskey, gin; \~40% alcohol).
* Have a history of alcohol or substance abuse in the 12 months prior to screening (including having been hospitalized for such in an in-patient or out-patient intervention program) or use that in the opinion of the investigator may be of a concern for the study.
* Reports a significant blood loss or blood donation totaling between 101 mL to 449 mL of blood within 30 days prior to baseline or a blood donation of more than 450 mL within 56 days prior to baseline.
* Reports donating plasma (e.g., plasmapheresis) within 15 days prior to baseline.
* Current enrolment or past participation in another study with any product(s) with at least one active ingredient within 30 days before first dose of study product or longer, if the previous test product is deemed by the investigator to have lasting effects that might influence the eligibility criteria or outcomes of current study.
* Living in the same household as another currently/previously enrolled participant in the present study.
* Any other medical condition/situation or use of medications/supplements/therapies that, in the opinion of the investigator, may adversely affect the participant's ability to participate in the study or its measures or pose a significant risk to the participant.

Nicotinamide mononucleotide (NMN) 600 mg/day administered orally as 3 capsules once daily for approximately 4 weeks.

Matching placebo administered orally as 3 capsules once daily for approximately 4 weeks.

Trigonelline monohydrate administered orally as 3 capsules once daily for approximately 4 weeks. Dose levels include 170 mg, 340 mg, 510 mg, and 680 mg trigonelline monohydrate, equivalent to 150 mg, 300 mg, 450 mg, and 600 mg trigonelline, respectively.

Participants receive 170 mg of trigonelline monohydrate (equivalent to 150 mg trigonelline) administered orally as 3 capsules once daily for approximately 4 weeks.

Participants receive 340 mg of trigonelline monohydrate (equivalent to 300 mg trigonelline) administered orally as 3 capsules once daily for approximately 4 weeks.

Participants receive 510 mg of trigonelline monohydrate (equivalent to 450 mg trigonelline) administered orally as 3 capsules once daily for approximately 4 weeks.

Participants receive 680 mg of trigonelline monohydrate (equivalent to 600 mg trigonelline) administered orally as 3 capsules once daily for approximately 4 weeks.

Participants receive 600 mg of nicotinamide mononucleotide (NMN) administered orally as 3 capsules once daily for approximately 4 weeks.

Participants receive matching placebo capsules administered orally as 3 capsules once daily for approximately 4 weeks.