Pilot Study of GLY-LOW Supplementation in Postmenopausal Women With Obesity

A combination of generally regarded as safe (GRAS) compounds named GLY-LOW, which included: alpha lipoic acid, pyridoxamine, nicotinamide, piperine and thiamine, were examined in pre-clinical experiments. GLY-LOW supplementation reduced caloric intake and increased insulin sensitivity in mice. In female mice, GLY-LOW supplementation reversed aging-related declines in female hormones. Studies in humans are needed to examine the feasibility, utility and efficacy of GLY-LOW supplementation in post-menopausal women with obesity toward improving aging-related impairments. The effect of GLY-LOW supplementation on these obesity and biological age-related impairments in post-menopausal adult female humans with obesity is unknown. We aim to translate the findings of GLY-LOW supplementation in animals to a cohort of healthy, postmenopausal females at birth with obesity by conducting a one-group, no-placebo comparer, pre post intervention clinical trial. Additionally, we propose to examine the specific effect of supplementation by GLY-LOW on biological aging via retina scan. The objectives of the proposed pilot study are:

I. Conduct a 6-month pilot study to examine the feasibility, utility and efficacy of GLY-LOW supplementation in a total of 40 postmenopausal female born adults \> 55 years with obesity (\> 30 BMI)

Ia. Examine alterations in self-reported caloric intake and the following health and biological aging, parameters prior to and after 6 months of GLY-LOW supplementation:

1. Self-reported Caloric Intake
2. Metabolic disease risk
3. Cardiovascular disease risk
4. Metabolic assessments
5. Hormones
6. Physical Function and Fitness
7. Muscular strength
8. Cognitive Function and Depression assessments
9. Systemic inflammation
10. Biological aging
11. Safety parameters (also every 2 months during the intervention; ECG at baseline and 2 months only) 12, Compliance measures (pill counts and interviews every 2 months during the intervention)

Study Population:

Healthy, postmenopausal (\> 1 year from last menstrual cycle) adult females (at birth), \> 55 years of age with obesity (≥ 25 BMI)

Objectives/Purpose of the Study:

Worldwide rates of obesity increased from 100 million to 764 million between 1980 and 2021. During the same time frame, a five-fold increase in Type 2 diabetes (T2DM) was reported4. Increases were similar across the globe with older individuals at greatest risk of obesity and related metabolic dysfunction. Central obesity is well known to be both a manifestation and driver of metabolic syndrome5 with similar prevalence worldwide6. Older, post-menopausal females, in particular, are at increased risk of developing central obesity due to a reduction in endogenous ovarian hormone production7. Novel multi-modal therapies are needed to address central obesity-related metabolic dysfunction in post-menopausal females. However, current therapeutic methods lack specificity and propose universal solutions to a multi-factorial disorder requiring an individualized, precision medicine approach8. More concerning, is the lack of rigorous, scientific evidence to support conventional dietary therapies for reducing aging-related central obesity and associated metabolic dysfunction9. Targeting multiple biological pathways that are related to individual behavioral determinants (caloric intake) and biological aging markers (estrogen levels) may identify more precise therapies in post-menopausal female adults with obesity.

Recently, advanced glycation end-products (AGEs) were identified as potential drivers of obesity-related impaired metabolic function10. In a series of in vitro and in vivo experiments, a combination of GRAS compounds that function synergistically to improve metabolic health and extend lifespan \[(alpha lipoic acid, pyridoxamine, nicotinamide, piperine and thiamine (i.e. GLY-LOW)\] were examined11-13. Supplementation with GLY-LOW was shown to detoxify methylglyoxal (MGO), a reactive precursor to AGEs. In addition, GLY-LOW supplementation reduced caloric intake and glycolysis, reprogramed metabolism, and increased insulin sensitivity and mTor signaling in the hypothalamus of the mice. In female animals GLY-LOW supplementation reversed aging-related declines in estrogen and related female hormones. Studies in humans are needed to translate these findings and explore the feasibility, utility and efficacy of GLY-LOW supplementation in post-menopausal women with obesity toward improving aging-related impairments. These include increased metabolic disease risk and insufficiency, impaired physical function, osteoporosis, reduced fitness levels, cognitive impairment, systemic inflammation and premature biological aging. Interestingly, results of a novel biological aging assessment, retina scan, were recently shown to be associated with metabolic dysfunction via inflammatory pathways14. The effect of GLY-LOW supplementation on these obesity and biological age-related impairments in post-menopausal adult female humans with obesity is unknown.

Study Site:

The Hoskinson Health and Wellness Clinic (HHWC) focuses on your whole health for your whole life, and provides a proactive, not reactive, personalized and integrated, multi-component solution to age-related increases in obesity and associated declines in metabolic health. The HHWC utilizes functional, precision medicine to identify unhealthy aging biomarkers and genetic vulnerabilities, and innovative medical technology to both diagnose and manage aging-related metabolic, physical and cognitive impairments. State-of-the-art assessments identify targets for tailored programming including diet instruction, nutrition education and supplementation, fitness counseling and training. The HHWC proposes to conduct a series of clinical studies using these state-of-the-art measures of aging-related parameters. Initially, we aim to translate the findings of GLY-LOW supplementation in animals to a cohort of healthy, postmenopausal females at birth with obesity by conducting a one-group, no-placebo comparer, pre post intervention clinical trial. Additionally, we propose to examine the specific effect of supplementation by GLY-LOW on biological aging via retina scan. The objectives of the proposed pilot study are:

I. Conduct a 6-month pilot study to examine the feasibility, utility and efficacy of GLY-LOW supplementation in a total of 40 postmenopausal female born adults \> 55 years with obesity (\> 25 BMI)

Ia. Examine alterations in self-reported caloric intake and the following health and biological aging, parameters prior to and after 6 months of GLY-LOW supplementation:

1. Self-reported Caloric Intake
2. Metabolic disease risk
3. Cardiovascular disease risk
4. Metabolic assessments
5. Hormones
6. Physical Function and Fitness
7. Muscular strength
8. Cognitive Function and Depression assessments
9. Systemic inflammation
10. Biological aging
11. Safety parameters (also every 2 months during the intervention; ECG at baseline and 2 months only)
12. Compliance measures (pill counts and interviews every 2 months during the intervention)

Trial Design:

A. The Primary Endpoint of the study is metabolic disease risk

C. Design of the Trial:

One group, no-placebo comparer, pre post intervention clinical trial

D. Randomization and Blinding:

The one group, no-placebo comparer pilot study will not require randomization. Study data managers and biostatisticians will be blinded to basic demographic and clinical characteristics (e.g. female, postmenopausal) of the study population and the overall purpose of the trial.

Initial Recruitment/Screening, Enrollment, and Baseline Evaluation:

Recruitment, screening and enrollment will occur during the first eight weeks of the study. Approximately 40 participants will be screened and enrolled into the study. Participation in the baseline evaluation will occur during weeks 9-12 (Table 1). Recruitment will be by referrals, personal interview, brochure mail-outs to area physicians, community center flyers and presentations, presentations at civic meetings, mass media announcements, social media posts, as well as clinic screenings as currently used in our existing studies and clinical services at the Hoskinson Health and Wellness Center. The plan is described below:

1. Potential study participants will be provided with a general information handout or link to our website regarding the study and contact information for participation.
2. Study personnel will receive phone, email inquiries or website inquiries and conduct a pre-screening interview and complete a checklist to determine eligibility.
3. Study personnel and/or other medical staff will then conduct the IRB-approved informed consent procedures and begin the screening process for study participants. An explanation of all procedures, potential risks, temporary side effects, anticipated benefits, and alternative methods (nutrition and fitness counseling, weight loss pharmaceuticals or nutraceuticals, other anti-aging therapeutics) will be given to the study participant. Confidentiality is assured as well as the right not to participate or to withdraw at any time. The signature of the study participant will be obtained on the consent form. A copy of the forms will be given to each study participant. Assurance that all questions have been answered about the study testing is obtained from the participants. Names and telephone numbers of contact persons on the medical team are given to each study participant for use if future questions arise.
4. General health screenings will then be performed by qualified medical staff. A medical physical evaluation to verify eligibility will be performed at the intervention baseline visits and a list of current medications will be required.
5. After clearance by the study physician, the study participant will receive the first set of baseline measurements The study coordinator will schedule the second day of baseline measurements with the study participant. The study participant will be advised to fast for at least 12 hours prior to the second day of baseline testing

E. Treatment, Dosage and Dosing regimen of the GLY-LOW supplement:

This supplement is available for over-the-counter purchase2. Each capsule is a combination of vitamins and natural products: Vitamin B1 (100mg); Vitamin B6 (50mg); Niacin (200mg); Alpha Lipoic Acid (150mg); and Piperine (15mg). Each participant will take this supplement daily in a pill form orally once in the morning. The test product will be two capsules a day with breakfast between 7:00 - 11:00 AM. The chosen doses were based on dose conversion from mice to humans and the prior data on safety for each of the compounds in humans.

F. Description of the Dosage Form, Packaging and Labeling of the GLY-LOW supplement:

G. Expected Duration of Subject Participation: 34 weeks (Figure 1 - Consort Diagram)

I. Available Alternative Treatments offered at the HHWC include nutrition and fitness counseling, weight loss pharmaceuticals or nutraceuticals, other anti-aging therapeutics. Study participants will be provided with information concerning these alternative treatment therapies during the consent procedures.

Follow-up Assessments:

1. The study participant will attend a follow-up safety assessment visit every two months and will participate in blood tests and MSQ. ECG will be assessed at baseline and 2 months only.
2. The study participants will attend one follow-up assessment after 6 months of receiving the study supplement (GLY-LOW). Baseline measurements will be repeated during these follow-up assessments in order to examine the effectiveness of the intervention.

Eligibility Criteria:
Inclusion Criteria:

* Post-menopausal (\>1 year from last menstrual cycle) adult females (at birth) \>55 years of age with obesity (BMI ≥ 25)

Exclusion Criteria:

* Males
* Adults not females at birth
* Adult females diagnosed with Gout
* Adult females receiving hormone replacement therapy (HRT)
* Adult females who are currently prescribed or received weight loss medications within the past 6 months, or currently enrolled in a defined weight loss program.
* Adult females with cardio-metabolic disease \[e.g. cardiovascular disease (CVD), type 2 diabetes (T2DM), hypertension, h/o stroke etc.)\] requiring any prescription medication.
* Adult females with other chronic immune, pulmonary, neurodegenerative or systemic disease requiring prescription medication
* Adult females with severe asthma based upon American Thoracic Society (ATS) standards and/or NIH guidelines.
* Adult females requiring monoclonal antibody or biological treatment
* Adult females diagnosed with an acute lower respiratory or GI infection within 2 weeks
* Adult females who are pregnant (would not qualify as post-menopausal) or nursing mothers
* Adult females reporting moderate to severe mental or physical disability
* Adult females reporting unwillingness to travel to onsite clinical facilities
* Adult females with moderate to severe cognitive impairment
* Adult females diagnosed with an eating disorder
* Adult females on blood thinners and anti-platelet drugs (other than aspirin), on chronic steroids (including inhaled), on spironolactone, on statins, on diuretics, on chronic pain medications, on any medication that affects glucose, insulin, lipids, metabolic performance
* Adult females with any recent change in chronic disease or any recent change in medication.
* Adult females on supplements with potential to augment or compete with the GLY-LOW.