Safety and Preliminary Efficacy of Aleeto in Amyotrophic Lateral Sclerosis

This study is a single-center, randomized, double-blind, placebo parallel-controlled, dose-escalation clinical study. The aim of this study was to evaluate the safety, tolerability, and preliminary effect of Aleeto in adult patients with ALS, and to provide an appropriate dose for the future clinical trial.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease, which is characterized by progressive loss of number and function of upper and lower motor neurons located in the brain and spinal cord, which leads to paralysis. ALS mainly begins in the limbs and face, where about one-third of patients have difficulty in speaking, chewing, or swallowing, then progressively affecting the pharyngeal muscles, attacking the sphincter in the advanced stages of the disease, and eventually affecting the trunk and respiratory function as the disease progresses. "Aleeto" derived from cellular exosomes, is a group of specific microenvironment protein polymers secreted by stem cells under stressed conditions, which has the advantages of selective assembly, targeted delivery, efficient repair of damaged tissues, high safety, stable chemical properties, and easy preservation. What's more, Aleeto has a strong nerve repair function. This study is a single-center, randomized, double-blind, placebo parallel-controlled, dose-escalation clinical study, aiming to evaluate the safety, tolerability, and preliminary effect of Aleeto in adult ALS patients, and provide an appropriate dose for the future clinical trial. In this study, we plan to enroll 24 patients, and will be randomly assigned to 4 groups. The initial dose group consists of four subjects, out of which three subjects will receive intravenous administration and intrathecal administration of 0.5μg/kg Aleeto, while one subject will receive a placebo. Fourteen subjects will be randomly assigned to the 1μg/kg and 2μg/kg dose groups to receive intrathecal administration combined with intravenous administration, with one subject in each dose group randomly receiving placebo treatment. Six subjects will be randomly assigned to the 2μg/kg dose group to receive intravenous administration only. Data are collected in face-to-face interviews at baseline and Day 14, 30, 37, 60, 67, 90, 120 follow-up visits. All patients will be examined before and after treatment, and the results will be compared. The primary outcomes are the incidence of adverse events (AEs) and serious adverse events (SAEs), and the incidence of abnormal changes in laboratory examination indicators, vital signs, neurological physical examination, and electrocardiogram within 120 days after the treatment. The secondary outcomes included changes in immunological indicators, pharmacokinetics, ALS Functional Rating Scale-Revised (ALSFRS-R), as well as the incidence of invasive mechanical ventilation and mortality rate . The statistical analysis was performed by bilateral test, and the test level α was 0.05. Baseline Equilibrium Analysis: Analysis of variance or Kruskal-Wallis H test is used to compare continuous data.The Chi-Squared test or Fisher's exact test is used to compare the categorical data. Primary Efficacy Analysis: The comparability among different dosage groups in terms of incidence of adverse events (abnormal liver functions, hypersensitivity reactions, liver failure, dyspnea, etc.), severe adverse events, and other abnormal indicators will be analyzed using Chi-squared tests or Fisher's exact test. Changes in laboratory examination indicators and vital signs before and after treatment will be analyzed using paired t-tests or Wilcoxon signed rank sum test. The intergroup differences in laboratory examination indicators and changes in vital signs before and after treatment will be analyzed using t-test or Mann-Whitney U test. Secondary Efficacy Analyses: The improvement of immunological indicators (lymphocyte subgroup analysis), pharmacokinetic indicators, and ALSFRS-R before and after treatment will be analyzed using paired t-tests or Wilcoxon signed rank sum test. The intergroup comparison of immunological indicators (lymphocyte subgroup analysis), pharmacokinetic indicators, and ALSFRS-R before and after treatment will be analyzed using t-tests or Mann-Whitney U tests. The intergroup significance of the incidence of invasive mechanical ventilation and mortality rate is determined using Chi-squared tests or Fisher's exact test. Exploratory Analysis: Comparison of measurement data: The improvement of ALSAQ-40, EQ-5D-5L, modified Norris scale, forced lung capacity, muscle strength, electromyographic indicators, gait function, biomarker levels, FSS, imaging indicators, HAMA and HAMD scales will be analyzed using paired t-tests or Wilcoxon signed rank sum test. The above exploratory analysis indicators and the difference in survival time between two groups will be analyzed using t-test or Mann-Whitney U test. Analysis of variance or Kruskal Wallis H-test will be used for comparing the differences ａｍｏｎｇ different dosａｇｅ groups, and Bonferroni correction method will be used for multiple comparisons. Mixed linear models will be used for analyzing the differences in the trend of changes between different dose groups over a period of 120 days (the data which corresponds to skewed distribution will be transformed). LSmeans will be used for analyzing the differences of indicators between different groups at different times, as well as the differences within the same group at different times. Comparison of enumeration data: Chi-square test or Fisher's exact test for enumeration data.