ZEO ScientifiX, Inc.

Background & Aim: Amniotic fluid (AF) has recently been characterized to contain a reserve of extracellular vesicles (EVs) with anti-inflammatory properties.The FDA has recently approved an investigational new drug application of an AF-derived EV biol. in a multicenter, phase I/II randomized, double blinded, placebo trial to evaluate the safety and potential efficacy of Zofin infused i.v. (IV) in patients experiencing prolonged COVID-19 symptoms (Long-Haulers).Methods, Results & Conclusion: Patient population (n = 30) includes those with symptoms consistent with COVID-19 infection for a prolonged period, greater than 6 wk and less than 12 mo.Patients must meet inclusion criteria requirements related to fatigue, extreme body aches, and joint pain.Symptoms must be persistent for a min. of 6 wk after COVID-19 recovery and neg. test.The investigational drug, Zofin, is an allogenic AF-derived EV biol. containing approx. 2.0 x 1011 nanoparticles/mL, 1.4 mg of protein/mL, and 300 ng of hyaluronic acid/mL.The treatment dose will be 1 mL at Day 0, 4, and 8 administered IV and diluted in 100 mL of saline.Patients will be randomized 1:1 into Zofin and placebo groups.The primary endpoint will be safety, where the incidence of grade 3 or 4 or serious adverse events in subjects receiving Zofin will be compared to placebo.Secondly, endpoints for efficacy will include various assessments such as COVID-19 associated symptoms, Fatigue Severity Score, Beck Depression Inventory, Mental Fatigue Questionnaire, pulse oxygen saturation, Heart Rate Variability, and transthoracic echocardiogram measures.Fifteen inflammatory biomarkers will be tested such as Plasma-CRP, D-dimer, TNF-a, IL-4, IL-13, IL-2, GM-CSF, sCD40L, CCL5 (RANTES), CCL3 (MIP-1a), IL-6, IL-10, IFN-g, VEGF, IL-8, and CCL4 (MIP-1b).The study duration will be 6 mo with follow-up at Days 0, 4, 8, 14, 21, 30, 60, 90, and 6 mo.Peripheral blood mononuclear cells will be collected for immune cell activation and response to Zofin.The incidence of adverse events will be characterized by type, severity, duration, and relationship to study meditation within 30 days related to the therapy.The changes in efficacy measurements will be compared to baseline.After unblinding, Zofin safety and potential efficacy will be determined from the placebo group comparison.Results of this trial will provide the first insights into an extracellular vesicle biol.'s effect in COVID-19 Long Haulers.

Extracellular vesicles (EVs) are being tested for their use as novel therapeutics. However, the optimal source of EVs is currently under investigation. Amniotic fluid (AF) is a natural source of EVs that can be easily obtained for use in regenerative medicine, yet AF-EV characterization has not been fully explored.

Here the authors demonstrate AF as a rich source of EVs and identify the microRNA and proteomic cargo. Bioinformatics analysis of this cargo revealed multiple pathway targets, including immunomodulatory, anti-inflammatory and free radical scavenging networks. The authors further demonstrated the therapeutic potential of this EV product as a novel preventative agent for bronchopulmonary dysplasia (BPD).

Intra-tracheal administration of AF-EVs preserved alveolar development, attenuated vascular remodeling and pulmonary hypertension, decreased lung pro-inflammatory cytokine expression and reduced macrophage infiltration in an experimental BPD model.

The authors' results suggest that AF is a viable biological fluid for EV harvest and that AF-EVs have strong therapeutic potential for pulmonary diseases, such as BPD, warranting further development to transition this novel EV product into the clinic.

Background & Aim: A human coronavirus (HCoV-19) has caused the novel coronavirus disease (COVID-19) outbreak worldwide.There is an urgent need to develop new interventions to suppress the excessive immune response, protect alveolar function, and reduce lung and systemic organ damage.Zofin is an acellular biol. that contains the soluble and extracellular vesicle fraction of human amniotic fluid.We have recently initiated single patient emergency/compassionate use eINDs for 12 subjects under our approved parent IND 19881.These trials included patients with various stages of COVID-19 infection and respiratory distress (mild to moderate ARDS, Post-COVID-19 complications, and Multi-organ failure in the ICU).Methods, Results & Conclusion: Zofin contains 2.3 x 1011 particles/mL with 70-80% pos. expression of exosome markers CD63 and CD81.Zofin was administered IV as 1mL doses (3-4 doses) within the first 3-4 visits.Scheduled visits were day 0,4,8,12,21, 28, and 60.If BMI was greater than 40, an addnl. visit was included at day 6 to administer an extra dose.The primary objective of these studies was to demonstrate the safety of Zofin while secondarily observing for efficacy by SOFA score assessment, chest X-Rays, and inflammatory biomarker testing.The approved FDA IND numbers are as follows: eIN-D#1-IND#22370, eIND#2-IND#22371, eIND#3-IND22897, eIN-D#5-IND#25426, eIND#11-IND#26776.The clin. outcome of each patients has varied, depending on the severity and baseline clin. status.However, we have found an overall pos. trend for a number of clin. improvements such as systemic inflammation (decreased CRP), improved organ failure scores for ICU patients, respiratory and fatigue improvements in outpatients.Importantly, there were no reported adverse events related to product administration or treatment in any of the treated patients.This is the first demonstration of allogenic, amniotic fluid-derived extracellular vesicles as a safe and potentially efficacious therapeutic treatment for respiratory failure induced by COVID-19 infection.