Evaluation of Metformin Hydrochloride on Endometrial Function in Women with Idiopathic Infertility: A Randomized, Controlled, Double-Blind Clinical Trial

The primary objective of this randomized, controlled, double-blind clinical trial is to evaluate the effect of **metformin** on endometrial function in women diagnosed with idiopathic infertility. This study aims to demonstrate that the proposed therapy can improve fertility in these women, which is clinically relevant as it addresses a significant challenge in reproductive medicine by potentially offering a therapeutic option for enhancing fertility outcomes in this patient population.

Secondary objectives include:

• Evaluation of the effect of metformin therapy on endometrial function by assessing the concentration of markers such as cytokines, IGF, LIF, TGF-α, EGF, HB-EGF, VEGF, histamine, inhibin B, relaxin, and insulin receptor in the tissue before and after treatment.
• Transcriptome analysis of endometrial tissues using next-generation sequencing (NGS) before and after metformin therapy.
• Intratissue metabolomic profiling of steroids to determine if metformin treatment affects the intracrine production of steroid hormones in the endometrium.
• Assessment of the effect of metformin treatment on the regulation of oxidative stress in the endometrium.
• Analysis of the Quality of Life (QoL) questionnaire of the patients undergoing the study and the analysis of dietary menus combined with the DXA study.

The clinical trial focuses on evaluating the effect of metformin on **endometrial function** in women diagnosed with **idiopathic infertility**. The study population consists exclusively of female participants, aged between 20 and 40 years, who have not achieved pregnancy for at least 12 months prior to screening. Participants are required to have a body mass index (BMI) ranging from 18.5 to 30 kg/m² and must be eligible for the in vitro fertilization program. The trial excludes individuals who have used hormone therapy or any methods of contraception within 30 days before screening and during the examination. The sponsor has not provided information regarding the total number of participants. The trial population was selected based on specific inclusion criteria, ensuring that only those diagnosed with idiopathic infertility and meeting the outlined health and lifestyle requirements are included. The study does not include male subjects and involves a vulnerable population, as defined by the trial's criteria.

The clinical trial is designed as a **randomized, controlled, double-blind** study to evaluate the effect of **metformin** treatment on endometrial function in women diagnosed with **idiopathic infertility**. The trial will involve the administration of **GLUCOPHAGE 500 mg** film-coated tablets, containing **metformin hydrochloride**, and a placebo. The study is set to commence on March 20, 2023, and is expected to conclude by January 31, 2028, with a maximum treatment period of 24 months. Participants will be randomly assigned to either the treatment group receiving metformin or the placebo group, ensuring the double-blind nature of the trial.

The trial will include several study visits, beginning with an inclusion (screening) visit to assess eligibility based on criteria such as age (20-40 years), a minimum of 12 months of infertility, and a BMI between 18.5 and 30 kg/m². Participants must not have used hormone therapy or contraception 30 days prior to screening and must be eligible for the in vitro fertilization program. Follow-up visits will be scheduled to monitor the primary endpoint of obtaining a pregnancy during the study and secondary endpoints, including the evaluation of endometrial function through blood tests and transcriptome analysis, as well as intrauterine metabolomic profiling and oxidative stress regulation.

The end-of-study visit will assess the overall outcomes and gather data on the quality of life and lifestyle factors impacting the effectiveness of in vitro fertilization. Participant involvement is expected to last up to 24 months, with conditions for early termination including non-compliance with study protocols or adverse reactions to the treatment. The trial aims to provide comprehensive insights into the potential of metformin to improve fertility outcomes in women with idiopathic infertility.

The clinical trial involves the administration of **metformin hydrochloride** as the experimental medication. The product used is marketed under the name **GLUCOPHAGE 500 mg** and is provided in the form of **film-coated tablets**. Each tablet contains 500 mg of the active substance, metformin hydrochloride, which is a chemical compound. The medication is administered orally, with a maximum daily dose of 1500 mg. The total maximum dose over the treatment period is 241,500 mg. The treatment duration is set for a maximum of 24 months. The pharmaceutical product is manufactured by Merck Santé S.A.S. and is authorized for use in Poland. Participant compliance with the dosing schedule will be monitored throughout the study.

The study also includes a **placebo** as a non-experimental treatment to serve as a comparator. The placebo is designed to mimic the appearance of the experimental medication but does not contain any active pharmaceutical ingredients. The placebo is used to maintain the double-blind nature of the trial, ensuring that neither the participants nor the investigators are aware of the treatment assignments. The placebo administration follows the same oral route and dosing schedule as the experimental medication to ensure consistency in the trial protocol.

The efficacy of the clinical trial will be assessed through a combination of primary and secondary endpoints. The primary endpoint is the achievement of pregnancy during the study period. Secondary endpoints include a comprehensive evaluation of the effect of **metformin** therapy on endometrial function, which will be assessed indirectly through blood tests measuring concentrations of specific markers such as cytokines (interleukin 1 and 2), insulin-like growth factor (IGF), leukemia inhibitory factor (LIF), transforming growth factor alpha (TGF-α), epidermal growth factor (EGF), heparin-binding growth factor-like EGF (HB-EGF), vascular endothelial growth factor (VEGF), histamine, inhibin B, relaxin, and insulin receptor.

Additionally, the study will utilize next-generation sequencing (NGS) technology to analyze the transcriptome of endometrial tissues, including a detailed microRNA profile, to assess the potential of microRNAs as predictive markers for the effectiveness of in vitro fertilization procedures. Intrauterine metabolomic profiling of steroids will also be conducted to identify potential prognostic markers for increasing pregnancy rates. The effect of metformin on the regulation of oxidative stress will be evaluated, considering its role in stimulating antioxidant processes and reducing reactive oxygen species (ROS) synthesis, which may enhance embryo implantation chances. Furthermore, the Quality of Life (QoL) questionnaire and dietary analysis combined with DXA examination will provide insights into the impact of diet and lifestyle on the effectiveness of in vitro fertilization, offering a holistic assessment of patients' health and well-being.