Evaluation of Oocyte/Embryo Cryopreservation Efficacy in Young Breast Cancer Patients Undergoing Adjuvant Chemotherapy with Follitropin Alfa and Follitropin Beta

The primary objective of this study is to evaluate the **efficiency** of oocyte or embryo cryopreservation following controlled ovarian stimulation in young patients with **breast cancer** who are scheduled to receive adjuvant chemotherapy. This is particularly assessed in terms of the number of Meta II oocytes that can be vitrified. This objective is clinically relevant as it addresses the potential impact of chemotherapy on fertility, providing insights into fertility preservation options for young breast cancer patients.

Secondary objectives include:

• Evaluating the proportion of patients with sufficient ovarian reserve eligible for controlled ovarian hyperstimulation.
• Assessing the toxicity of controlled ovarian hyperstimulation, focusing on the risk of ovarian hyperstimulation syndrome (OHSS) and vascular complications such as thrombosis.
• Determining the impact of the fertility preservation program on the schedule of anticancer treatment.
• Evaluating the gonadotoxicity of chemotherapy by monitoring AMH levels, AFC over time, and the incidence and duration of chemotherapy-induced amenorrhea, premature ovarian failure, and definitive menopause.
• Assessing fertility and fecundity in terms of cumulative incidence of pregnancy and live birth, and describing the type and outcome of pregnancies.
• Measuring the degree of project completion of subsequent pregnancies in patients who started controlled ovarian hyperstimulation and expressed a wish to be pregnant.
• Assessing the number of patients wishing to reuse their frozen gametes.
• Evaluating the long-term safety of the procedure in terms of the risk of relapse compared to historical controls.
• Identifying new non-invasive biomarkers of follicle/oocyte quality based on miRNA and cfDNA in patients undergoing controlled ovarian hyperstimulation.

The clinical trial focuses on **breast cancer in young women**, specifically evaluating the efficiency of oocyte or embryo cryopreservation after controlled ovarian stimulation in patients who will receive adjuvant chemotherapy. The study population comprises women aged 18 to 38 years, all of whom have been newly diagnosed with histologically proven breast cancer. Participants are required to have no prior chemotherapy and must be affiliated with a public health insurance program. The trial exclusively involves female subjects, with no male participants included. The sponsor has not provided information regarding the total number of participants. Key lifestyle considerations, such as diet or physical activity, are not specified. The trial does not involve a vulnerable population. Participants must have a sufficient ovarian reserve, as indicated by specific hormonal and follicular criteria, and must have signed an informed consent form. The trial population was selected based on these criteria, ensuring a focus on young women who are planning to undergo standard sequential anthracycline and taxane-based chemotherapy, with potential inclusion of Trastuzumab and Hormonotherapy, according to local practices.

The clinical trial is designed to evaluate the efficiency of **oocyte** or embryo cryopreservation following controlled ovarian stimulation in young women diagnosed with breast cancer who are scheduled to receive adjuvant chemotherapy. This study is a **randomized**, **controlled**, and **double-blind** trial, categorized as a Phase 4 study, with an estimated duration from December 2016 to June 2026. Participants will be randomly assigned to receive either **follitropin alfa** or **follitropin beta**, both administered via subcutaneous injection, with a maximum daily dose of 450 IU and a total dose not exceeding 9000 IU over a 20-day treatment period.

The trial will commence with an inclusion (screening) visit to confirm eligibility based on criteria such as age (18-38 years), newly diagnosed breast cancer, planned adjuvant chemotherapy, and sufficient ovarian reserve. Following the screening, participants will undergo controlled ovarian stimulation, with study visits scheduled to monitor ovarian response and retrieve oocytes. The primary endpoint is the number of Meta II oocytes that can be vitrified, while secondary endpoints include the total number of embryos and oocytes preserved, the rate of top-quality embryos, and the safety of study procedures.

Participants are expected to be involved in the study for the duration of the ovarian stimulation and cryopreservation process, with follow-up visits to assess the impact on ovarian reserve and the safety of the procedures. The end-of-study visit will evaluate the long-term outcomes, including the impact on fertility preservation and the schedule of anti-cancer treatment. Conditions that may lead to early termination from the study include non-compliance with the protocol, adverse events, or withdrawal of consent. The trial aims to provide valuable insights into fertility preservation strategies for young breast cancer patients undergoing chemotherapy.

The clinical trial involves the administration of **Follitropin Beta**, a medication of biological/biotechnological origin, used for controlled ovarian stimulation. The pharmaceutical form is denoted as PHF00231MIG. The medication is administered via **subcutaneous use**. The maximum daily dose is 450 IU international unit(s), with a total maximum dose of 9000 IU international unit(s) over a treatment period not exceeding 20 days. The administration schedule is designed to ensure optimal ovarian response, and participant compliance is monitored through regular follow-ups and dose adjustments as necessary.

Additionally, the trial includes the use of **Follitropin Alfa**, another biological/biotechnological product, which shares the same pharmaceutical form, PHF00231MIG, and administration route, subcutaneous use. The dosing regimen for Follitropin Alfa is identical to that of Follitropin Beta, with a maximum daily dose of 450 IU international unit(s) and a total maximum dose of 9000 IU international unit(s) over a 20-day period. The trial does not involve any pediatric formulations or orphan drug designations for either medication. Compliance with the dosing schedule is critical and is monitored through participant logs and clinical assessments.

No non-experimental treatments, such as standard-of-care therapy, placebo, or comparator treatments, are specified in this trial. The focus remains on evaluating the efficiency of oocyte or embryo cryopreservation following controlled ovarian stimulation in young breast cancer patients undergoing adjuvant chemotherapy. The trial aims to assess the number of Meta II oocytes that can be vitrified, providing valuable insights into fertility preservation strategies for this patient population.

Efficacy in this clinical trial will be assessed primarily by evaluating the quality of oocyte retrieval, specifically in terms of the number of **Meta II oocytes** that can be vitrified. Secondary endpoints will include the total number of embryos and oocytes preserved, the rate of top-quality embryos, and the number of mature, immature, and atretic oocytes. The ovarian reserve at study entry will be described using **AMH** levels and Antral Follicle Count (AFC), with eligibility for controlled ovarian stimulation determined by an AMH level of at least 6 pmol/l or an AFC of at least 6 follicles.

Additional secondary endpoints will assess the safety of study procedures over the month following egg retrieval, using the NCI-CTCAE scale, version 4.0, and evaluating clinical, ultrasound, and biological signs. The impact of the fertility preservation program on the schedule of anti-cancer treatment will be measured by the time interval between surgery and the start of chemotherapy, with a delayed start defined as more than 60 days post-surgery. Gonadotoxicity of chemotherapy will be evaluated by AMH levels, AFC, and chemotherapy-induced amenorrhea, with premature ovarian failure and definitive menopause defined by specific time intervals from the last menstrual period.

Pregnancy outcomes, including the type of pregnancy and outcome, will be reported over a 10-year period post-chemotherapy or until the patient reaches 43 years of age. Disease-free survival will be measured from the date of surgery to the first relapse or death, with data censored at the last follow-up for patients alive and disease-free. Optional translational research will involve quantifying circulating nucleic acids, including miRNAs and cell-free DNA, using quantitative real-time PCR techniques.