Supplementation for Male Subfertility (FertEnhancer)

Nutraceutical Supplementation for Male Subfertility

Old age, obesity, physical inactivity, environmental factors and genetics may contribute negatively to fertility in both males and females. In males, specifically, certain supplements, such as single antioxidants and trace minerals, have previously been shown to improve sperm function marginally. One hypothesis is that sperm function can be improved even further by combining several different types of supplements (e.g., amino acids, energy carriers, vitamins, antioxidants, and trace minerals) to target several age-related cell pathways, for example, oxidative stress, mitochondrial dysfunction, inflammation and cell energetics. This 3-month placebo-controlled, randomized clinical trial, aims to test the effects of a novel multi-ingredient supplement (Fertility Enhancer) that targets several age-related cell pathways on sperm function in overweight or obese and subfertile males.

Study Overview

• Status: Not yet recruiting
• Conditions: Male Infertility
• Intervention / Treatment: Dietary supplement: Active multi-ingredient supplement (Fertility Enhancer, FE); Dietary supplement: Inactive placebo (Placebo; PLA)

Detailed Description

BACKGROUND: Infertility is characterized by the failure to become pregnant after one year of regular intercourse without the use of contraceptives and impacts 10-15% of couples worldwide. Both male and female partners contribute to a couple's reproductive health, with approximately one third of infertility cases caused by male factors, one third by female factors, and the remaining by either a combination of both or unknown causes. The prevalence of infertility is a growing concern in Canada, as is seen in an increased use of assisted reproductive technology (ART), which may be both invasive and expensive. Cost-effective, safe, and accessible alternatives to ART are therefore needed. The most common cause of subfertility is 'biological aging', characterized by the hallmarks of aging, such as mitochondrial dysfunction, oxidative damage and inflammation. Another common cause of male and female subfertility is obesity, which is associated with multisystemic oxidative damage and inflammation.

PURPOSE: The aim of this placebo-controlled, double-blind randomized clinical trial is to test the effects of a multi-ingredient supplement (Fertility Enhancer) designed to target several aging- and obesity-related pathways on World Health Organization (WHO) semen quality parameters in overweight and obese and subfertile males (sperm count, motility, morphology and vitality).

SAMPLE-SIZE ESTIMATE AND DESIGN: Sperm count/concentration is strongly correlated to all World Health Organization semen quality parameters. With significance set at 0.05 (Z = 1.96) and power to 0.8 (Z = 0.84), a sample-size of 17-32 per group is sufficient to detect an increase of 10 x 10^6 spermatozoa/mL with a standard deviation of 15 to 20 x 10^6 spermatozoa/mL. Thus, sixty-four (n = 64) males between 25 and 50 years of age that are confirmed overweight or obese and subfertile will be randomized into age-matched Placebo (PLA, n = 32) vs Fertility Enhancer (FE, n = 32) groups and undergo daily supplementation for 3 months.

SUPPLEMENTS: The FE supplement contains energy carriers (creatine), conditionally essential amino acids (arginine), Omega 3 fatty acids (DHA and EPA), vitamins (B9, B12, E, and D3), antioxidants (CoQ10 and alpha lipoic acid), trace minerals (selenium, iron, zinc, and copper), and plant extracts (beet root, green tea, and green coffee bean). The isocaloric and inactive placebo contains safflower oil, microcrystalline cellulose and sugar and is identical in flavor to FE.

CO-PRIMARY OUTCOMES: All outcomes will be measured at baseline and post intervention for assessing % pre-to-post changes. Co-primary outcomes are body composition by dual x-ray absorptiometry, including lean mass to fat mass ratio (body composition index; BCI) and total fat mass, and the WHO semen quality parameters; specifically, % improvements in sperm count, motility, morphology, and vitality.

SECONDARY OUTCOMES: Secondary outcomes are % improvements in sperm DNA fragmentation (flow cytometry-assessed) and markers of oxidative damage (protein carbonyls, lipid peroxidation, 8-hydroxydeoxyguanosine)), inflammation (interleukin-1, tumor necrosis factor-alpha, interleukin-6), apoptosis (total and cleaved caspase 3), cell cycle arrest (p16 and p21), mitochondrial biogenesis (complexes I-V), antioxidant status (superoxide dismutases 1 and 2), and energy state (ATP and phosphocreatine).

OTHER: Additional outcomes are body morphology (bodyweight, waist/height ratio, and body mass index), other body composition outcomes (lean mass and appendicular skeletal muscle mass index), and blood markers of oxidative damage (malondialdehyde), inflammation (c-reactive protein, interleukin-1, tumor necrosis factor-alpha, interleukin-6), antioxidant status (ORAC, TEAC), liver enzymes (alanine aminotransferase, aspartate aminotransferase, and creatinine) and energy state (ATP & phosphocreatine levels).

HYPOTHESIS: The main hypothesis of the current trial is that co-primary body composition outcomes and the World Health Organization (WHO) semen quality parameters (count, motility, morphology, and/or vitality) will be significantly improved following FE supplementation and superior to PLA.

STATISTICS: A standard omnibus one-way repeated measures ANOVA F-test followed by Duncan post hoc analyses will be used for all parametric data analyses. Non-parametric equivalents will be used for non-normally distributed data with significance set at p = 0.05. Delta pre-post changes (% improvements) for all outcomes within and between groups are biologically relevant and planned a priori comparisons.

• Study Type: Interventional
• Enrollment (Estimated): 64
• Phase: Phase 2

Contacts and Locations

• Study Contact: Name: Mark A Tarnopolsky, PhD; Phone Number: 76593 9055212100; Email: tarnopol@mcmaster.ca
• Study Contact Backup: Name: Mats Nilsson; Phone Number: 76680 9055212100; Email: mats.nilsson@exerkine.com

Study Locations

• Canada
  • Ontario
    • Hamilton, Ontario, Canada, L8N 3Z5
      • Mark Tarnopololsky
      • Contact: Mark A Tarnopolsky, PhD; Phone Number: 76593 905-525-2100; Email: tarnopol@mcmaster.ca
      • Contact: Mats I Nilsson, PhD; Phone Number: 76680 905-525-2100; Email: mats.nilsson@exerkine.com

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Males between the ages of 25-50 years diagnosed with subfertility through Ontario Networks of Experts in Fertility (ONE Fertility, Burlington, ON).
• For diagnosis of male subfertility, the 2010 and 2021 World Health Organization criteria will be used for sperm count, motility, morphology and vitality.
• Overweight and obese males according to body mass index (BMI) between the ages of 25-50 years.

Exclusion Criteria:

• Smoking,
• history and drug alcohol abuse,
• BMI > 30 kg/m2,
• genital disease (cryptorchidism, current genital inflammation, or varicocele),
• genital trauma or surgery to the male reproductive system,
• known Y chromosome microdeletions or karyotype abnormalities (if known prior),
• hepatobiliary disease,
• significant renal insufficiency,
• occupational exposures to reproductive toxins,
• endocrine abnormality,
• recent or current sexually transmitted infection,
• use of cytotoxic drugs,
• use of immunosuppressants,
• use of anticonvulsants,
• use of androgens or antiandrogens,
• history of central nervous system injury,
• neurological or psychiatric disease to potentially compromise study data collection,
• treatment of erectile dysfunction with any drugs during the past 4 weeks,
• history of cancer chemotherapy,
• current supplementation with ingredients being tested unless 1-month washout period

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Triple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Active multi-ingredient supplement (Fertility Enhancer; FE); Volunteers will be randomized in a double-blinded fashion into the experimental treatment group, which entails daily supplementation of an active multi-ingredient supplement designed to enhance fertility (Fertility Enhancer; FE) for 3 months.	Dietary supplement: Active multi-ingredient supplement (Fertility Enhancer, FE); Consuming a multi-ingredient supplement targeting multiple cell pathways daily for 3 months.
Placebo Comparator: Inactive placebo (Placebo; PLA); Volunteers will be randomized in a double-blinded fashion into a placebo group, which entails daily supplementation of a calorie-matched, inactive placebo (Placebo; PLA) identical in flavor to the active supplement for 3 months.	Dietary supplement: Inactive placebo (Placebo; PLA); Consuming an inactive placebo that is calorie-matched to the active supplement daily for 3 months.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Percent change in sperm count/concentration from baseline to 3 months	Sperm count/concentration (millions spermatozoa/mL semen)	Baseline to 3 months
Percent change in sperm motility from baseline to 3 months	Proportion motile sperm (%)	Baseline to 3 months
Percent change in sperm morphology from baseline to 3 months	Proportion normal sperm morphology (%)	Baseline to 3 months
Percent change in sperm vitality from baseline to 3 months	Proportion viable sperm (vitality) (%)	Baseline to 3 months
Percent change in body composition index from baseline to 3 months	Lean mass/fat mass ratio by dual X-ray absorptiometry scan (body composition index; % change)	Baseline to 3 months
Percent change in total fat mass from baseline to 3 months	Total fat mass by dual X-ray absorptiometry scan (kg; % change)	Baseline to 3 months

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Percent change in sperm DNA fragmentation index from baseline to 3 months	Sperm DNA fragmentation index by flow cytometry (%)	Baseline to 3 months
Percent change in sperm DNA 8-hydroxydeoxyguanosine from baseline to 3 months	Sperm DNA 8-hydroxydeoxyguanosine by ELISA (ng/mL; %)	Baseline to 3 months
Percent change in sperm protein carbonyls from baseline to 3 months	Sperm protein carbonyls immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm lipid peroxidation (4-hydroxynonenal) from baseline to 3 months	Sperm 4-hydroxynonenal immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm antioxidant marker superoxide dismutase 1 from baseline to 3 months	Sperm superoxide dismutase 1 expression immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm antioxidant marker superoxide dismutase 2 from baseline to 3 months	Sperm superoxide dismutase 2 expression immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm apoptotic marker cleaved caspase 3 from baseline to 3 months	Sperm cleaved caspase 3 expression immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm apoptotic marker total caspase 3 from baseline to 3 months	Sperm total caspase 3 expression immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm mitochondrial OXPHOS from baseline to 3 months	Sperm mitochondrial OXPHOS expression immunoblot (optical density; %)	Baseline to 3 months
Percent change in sperm cell cycle arrest marker p16 from baseline to 3 months	Sperm p16 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm cell cycle arrest marker p21 from baseline to 3 months	Sperm p21 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammatory marker interleukin 1 from baseline to 3 months	Sperm interleukin 1 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammatory marker TNF-alpha from baseline to 3 months	Sperm TNF-alpha messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammatory marker interleukin-6 from baseline to 3 months	Sperm interleukin-6 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammatory marker interleukin-8 from baseline to 3 months	Sperm interleukin-8 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammatory marker interleukin-18 from baseline to 3 months	Sperm interleukin-18 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm inflammasome marker caspase 1 from baseline to 3 months	Sperm caspase 1 messenger RNA levels by rtPCR (fold control/placebo; %)	Baseline to 3 months
Percent change in sperm ATP levels from baseline to 3 months	Sperm ATP levels by ELISA (pM/100 mg protein; %)	Baseline to 3 months
Percent change in sperm phosphocreatine levels from baseline to 3 months	Sperm phosphocreatine levels by ELISA (ng/100 mg protein; %)	Baseline to 3 months

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Percent change in inflammatory cytokine TNF-alpha from baseline to 3 months	Serum TNF-alpha levels (pg/mL)	Baseline to 3 months
Percent change in bodyweight from baseline to 3 months	Bodyweight by standard scale (kg; %)	Baseline to 3 months
Percent change in body mass index from baseline to 3 months	Body mass index (BMI) (bodyweight/height squared; kg/m2; %)	Baseline to 3 months
Percent change in lean mass from baseline to 3 months	Lean mass by dual X-ray absorptiometry scan (kg; %)	Baseline to 3 months
Percent change in appendicular skeletal muscle mass from baseline to 3 months	Appendicular skeletal muscle mass by dual X-ray absorptiometry scan (kg; %)	Baseline to 3 months
Percent change in appendicular skeletal muscle mass index from baseline to 3 months	Appendicular skeletal muscle mass index by dual X-ray absorptiometry scan (kg/height squared; %)	Baseline to 3 months
Percent change in liver enzyme ALT from baseline to 3 months	Serum alanine aminotransferase levels (IU/L; %)	Baseline to 3 months
Percent change in liver enzyme AST from baseline to 3 months	Serum aspartate aminotransferase levels (IU/L; %)	Baseline to 3 months
Percent change in liver enzyme creatinine from baseline to 3 months	Serum creatinine levels (mg/dL; %)	Baseline to 3 months
Percent change in malondialdehyde levels from baseline to 3 months	Plasma malondialdehyde levels (uM; %)	Baseline to 3 months
Percent change in Oxygen Radical Absorbance Levels (ORAC) from baseline to 3 months	Plasma Oxygen Radical Absorbance Levels (ORAC units; %)	Baseline to 3 months
Percent change in Trolox Equivalent Antioxidant Capacity (TEAC) from baseline to 3 months	Serum Trolox Equivalent Antioxidant Capacity (mM; %)	Baseline to 3 months
Percent change in inflammatory cytokine interleukin-1 from baseline to 3 months	Serum interleukin 1 levels (pg/mL; %)	Baseline to 3 months
Percent change in inflammatory cytokine interleukin-6 from baseline to 3 months	Serum interleukin-6 levels (pg/mL; %)	Baseline to 3 months
Percent change in inflammatory marker c-reactive protein from baseline to 3 months	Serum c-reactive protein levels (mg/dL; %)	Baseline to 3 months
Percent change in ATP levels from baseline to 3 months	Plasma ATP levels (mmol/L; %)	Baseline to 3 months
Percent change in phosphocreatine levels from baseline to 3 months	Plasma phosphocreatine levels (mmol/L; %)	Baseline to 3 months

Collaborators and Investigators

• Sponsor: Hamilton Health Sciences Corporation
• Collaborators: One Fertility

Study record dates

Study Major Dates

• Study Start (Estimated): January 1, 2025
• Primary Completion (Estimated): February 2, 2026
• Study Completion (Estimated): February 2, 2026

Study Registration Dates

• First Submitted: October 16, 2023
• First Submitted That Met QC Criteria: October 16, 2023
• First Posted (Actual): October 23, 2023

Study Record Updates

• Last Update Posted (Actual): October 10, 2024
• Last Update Submitted That Met QC Criteria: October 8, 2024
• Last Verified: October 1, 2024

More Information

Terms related to this study

• Keywords: Antioxidants; Oxidative stress; Inflammation; Obesity; DNA; Infertility; Weight loss; Aging; Mitochondria; Creatine; Supplements; Arginine; Vitamins; ATP; Sub-fertility; Fragmentation
• Additional Relevant MeSH Terms: Urogenital Diseases; Male Urogenital Diseases; Genital Diseases, Male; Genital Diseases; Infertility; Infertility, Male
• Other Study ID Numbers: 16502

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: The study protocol, informed consent form and statistical analysis plan may be made available from February 2025 onward. This may occur along with the release of the deidentified results for publishing requirements and/or upon request by qualified researchers.
• IPD Sharing Time Frame: The study protocol, informed consent form and statistical analysis plan may be made available from February 2025 onward. This may occur along with the release of the deidentified results for publishing requirements and/or upon request by qualified researchers.
• IPD Sharing Access Criteria: Access to trial data can be requested by qualified researchers engaging in independent scientific research studies.
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No