Evaluation of Autologous Bone Marrow-Derived Mononuclear Cells and Saline in Advanced Chronic Limb Ischemia with High Amputation Risk

The primary objective of this study is to evaluate the **safety** and **efficacy** of autologous bone marrow-derived mononuclear cells (BM-MNCs) in the treatment of "no option" chronic limb ischemia, specifically in patients with advanced chronic ischemia (Fontaine III-IV class and Rutherford IV-V) who are at high risk of amputation. The study aims to assess safety by the absence of adverse effects and efficacy by measuring pain reduction, functional improvement, and the delay or absence of amputation. Additionally, the study seeks to determine the most efficient route of administration for BM-MNCs. This is clinically relevant as it addresses a critical need for therapeutic options in patients with severe limb ischemia who have limited treatment alternatives.

The clinical trial involves participants diagnosed with **advanced chronic ischemia** (Fontaine III-IV class and Rutherford IV-V), with a risk of amputation of at least one limb. The study population includes both male and female subjects, aged between 18 and 90 years. Participants are required to have no option for surgical or endovascular revascularization and a life expectancy greater than two years. They must also have normal hematological parameters, including leukocytes ≥ 3000, neutrophils ≥ 1500, and platelets ≥ 140,000. The trial does not involve a vulnerable population. The sponsor has not provided information regarding the total number of participants. Selection criteria include the ability to provide signed informed consent. Lifestyle factors such as diet, physical activity, or habits are not specified as part of the trial's considerations.

The clinical trial is designed to evaluate the **safety** and **efficacy** of autologous bone marrow-derived cells in the treatment of advanced chronic ischemia, specifically in patients with a high risk of limb amputation. This study is a Phase 4, randomized, double-blind, controlled trial. The trial aims to assess the absence of adverse effects and improvements in pain, functional capacity, and the delay or prevention of amputation. The trial is expected to run from May 2021 to April 2025, with participant involvement lasting up to three months.

Participants will undergo a series of study visits, beginning with an inclusion (screening) visit to confirm eligibility based on criteria such as age (18-90 years), advanced chronic ischemia classification, lack of surgical or endovascular revascularization options, and normal hematological parameters. Following the screening, eligible participants will be randomized to receive either the investigational treatment or a control. The investigational treatment involves the intra-arterial administration of bone marrow nuclear cells, with **saline** used as a solution for injection.

Throughout the trial, participants will attend follow-up visits to monitor safety and efficacy outcomes, including changes in the ankle/brachial index, digit/brachial index, transcutaneous oxygen measurement, ischemic ulcer size, and maximum walking distance until claudication. The end-of-study visit will conclude the participant's involvement, during which final assessments will be conducted. Conditions that may lead to early termination from the study include the occurrence of significant adverse effects or withdrawal of consent by the participant. The trial's primary endpoint focuses on anatomical and functional changes induced by the therapy, while secondary endpoints are not specified in the available data.

The clinical trial involves the use of **SALINE** as an experimental medication. **SALINE** is administered in the form of a **solution for injection**. The route of administration is **intraarterial use**, which involves delivering the solution directly into the arteries. The maximum daily dose of **SALINE** is set at 1 unit, with a total maximum dose of 3 units over the course of the treatment. The treatment period is limited to a maximum of 3 days. **SALINE** is not a pediatric formulation and is not classified as an orphan drug. The role of **SALINE** in this trial is to serve as a test substance.

In addition to the experimental treatment, the study also involves the use of **bone-marrow nuclear cells**. These cells are utilized as a non-experimental treatment in the study. The objective of the trial is to evaluate the safety and efficacy of these cells in the treatment of chronic arterial limb ischemia in patients at high risk of amputation. The study aims to assess the absence of adverse effects, pain reduction, functional improvement, and the delay or absence of amputation. The trial also seeks to determine the most efficient route of administration for the treatment.

The efficacy of the clinical trial will be assessed by evaluating the primary endpoints, which include anatomic and functional changes resulting from the vasculo/angiogenesis induced by somatic cell therapy. These changes will be measured through improvements in the ankle/brachial index, digit/brachial index, transcutaneous oxygen measurement, ischemic ulcer size, and maximum walking distance until claudication. The trial aims to determine the efficacy of bone marrow-derived mononuclear cells (BM-MNCs) in the treatment of chronic arterial limb ischemia in patients at high risk of amputation. The main objective is to assess pain decrease, functional improvement, and the delay or absence of amputation.