Efficacy and Safety of Ovotransferrin in COVID-19 Patients

Efficacy and Safety of Ovotransferrin in COVID-19 Patients With Mild-to-Moderate Disease

Chicken egg consists of three main components, egg white (56%), yolk (33%) and shell (11%). The key component of egg white is 88 percent water and 12 percent solids, of which over 90 percent are protein. Ovalbumin, ovotransferrin, and ovomucoid are considered the main proteins, while ovomucin, lysozyme, ovoglobulin, ovomacroglobulin, ovoglycoprotein, ovoflavoprotein, thiamin-binding proteins, ficin/papain inhibitors, avidin, and cystatin are the minor white proteins of the embryo. These proteins and their peptides have been recognized as antioxidants, antimicrobials, metal-chelators, anti-viral, anti-tumor, and angiotensin-converting enzyme (ACE)-inhibitors for their functional significance SARS-CoV-2 virus activates the human cell ACE2 receptor, triggering a series of deleterious events. In COVID19, renin-angiotensin is upregulated and the pathway is overexpressed and a progressive cytokine storm is always observed. In all these pathogenic processes, ovotransferrin could play a modifier function to enhance the condition. Ovotransferrin can be beneficial to the antioxidant effects of Angiotensin II by inhibiting NF-kB. Ovotransferrin is an immunomodulator that downregulates pro-inflammatory cytokines including TNF-α, IL-6, and IL-1 that could extinguish the cytokine storm. Ovotransferrin can also serve as a protective agent against COVID19 cardiotoxicity due to disruption in the ACE2-mediated signaling pathway, cytokine storm, pulmonary dysfunction, and side effects of medications.

In patients with coronavirus infection, provided ovotransferrin's possible protective effects, it is suggested as a supportive and therapeutic alternative.

Given this background, in the light of the current COVID-19 emergency, it is the intention of the investigators to conduct a prospective, randomized, open-label, controlled study in the cohort of hospitalized patients with covid-19 pneumonia, administering 200 mg of ovotransferrin orally in addition to the standard of care therapy (SOC).

The investigators hypothesize that the use of ovotransferrin will be associated with an earlier improvement of clinical and humoral parameters after 10 days of ovotransferrin treatment when compared to the group of patients provided with standard care.

Study Overview

• Status: Unknown
• Conditions: Covid19
• Intervention / Treatment: Dietary supplement: Ovotransferrin

Detailed Description

Chicken egg consists of three main components, egg white (56%), yolk (33%), and shell (11%). The key component of egg white is 88 percent water and 12 percent solids, of which over 90 percent are protein. Ovalbumin, ovotransferrin, and ovomucoid are considered the main proteins, while ovomucin, lysozyme, ovoglobulin, ovomacroglobulin, ovoglycoprotein, ovoflavoprotein, thiamin-binding proteins, ficin/papain inhibitors, avidin, and cystatin are the minor white proteins of the embryo. These proteins and their peptides have been recognized as antioxidants, antimicrobials, metal-chelators, anti-viral, anti-tumor, and angiotensin-converting enzyme (ACE)-inhibitors for their functional significance. Among these functional properties, the antioxidant ability is essential to food processing. Generation of reactive oxygen species (ROS) and free radicals is an integral part of biological processes affecting all living organisms in energy production. Ovotransferrin (OTf), which constitutes 12% of the total white egg protein, is known as an iron binder and transporter in animals and can thus be used as an iron replacement, antimicrobial or antioxidant. Japanese researchers find ovotransferrin in a redox-dependent method generates observable self-cleaning.

OTf is composed of 686 amino acids and has a molecular mass of 77.90 kDa. It is a superoxide dismutase (SOD)-mimicking protein with a potent superoxide anion (O2 _-)-scavenging activity and showed dramatically higher scavenging activity than some known antioxidants, such as serum albumin or ascorbate. Interestingly, metal-bound OTfs such as Fe2+-OTf, Mn2+-OTf, Cu2+-OTf exhibited greater O2 _- dismutation capacity than the apoprotein. Conjugation of OTf with small molecules such as catechin shows improved antioxidant activity. During in vitro experiments, OTf displayed some antioxidant effects and its hydrolysates showed approximately 3,2 to 13,5 times higher superoxide anion scavenging activity and better copper/calcium scavenging activity than OTf. Furthermore, autocleaved OTf showed higher antioxidant activity than the natural OTf, and the hydrolysates of autocleaved OTf showed stronger antioxidant activity than those of the autocleaved one.

The peptide IRW, resulting from hydrolysis, exhibited a high-oxygen radical-scavenging that could be related to tryptophan. In addition, two high antioxidant activity tetrapeptides (WNIP and GWNI) were found in OTf thermolysin digests. WNI's motif seemed to be responsible for the high antioxidant potential as amino acid residues coupled with both peptides' N or C terminus decreased their antioxidant capacity. Moreover, when tested in endothelial cells, the peptide GWNI showed the ability to reduce reactive oxygen species generation. Other antioxidant peptides previously reported from OTf using the oxygen radical absorbance ability, on the other hand showed no antioxidant activity in cells, showed deficiencies in cell-free in vitro methods for antioxidant studies, and highlighted the need to use more biological systems such as culture cells to test antioxidant peptides. OTf-derived hydrolyzates obtained with HCl at pH 2.5 or other enzymes such as protamex, alkalase, trypsin, neutrase, flavorzyme, maxazyme, collupulin, protex, promod 278 and alpha-chymotrypsin showing higher superoxide anion scavenging activity and radical oxygen absorption ability than intact protein and showing protective effects against oxidative stress-induced DNA.

Due to their anti-inflammatory and antioxidant properties, otrf-derived peptides, including IRW, IQW, and KVREGT, are viable treatment agents for endothelial dysfunction and CVD prevention. They de-regulate the expression of the cytokine-induced inflammatory protein in the vascular endothelium by modulating the pathway NF-μB. Recent research also revealed that KVREGT has a significant role to inhibit ACE and promote vasodilation. IQW impedes ACE, promoting antioxidation.

Ovotransferrin and lactoferrin are iron-binding proteins with natural immunity-related antiviral and antibacterial activities, displaying marked sequence and structural homologies. Two hen ovotransferrin fragments DQKDEYELL (hOtrf219-227) and KDLLFK (hOtrf269-301 and hOtrf633-638) have registered antiviral activity against Marek virus infection of chicken embryo fibroblasts. These fragments have sequence homology with two bovine lactoferrin fragments with antiviral activity towards herpes simplex virus, indicating that these fragments may play a role in exploiting intact protein antiviral activity towards herpes viruses. NMR analysis showed that these chemically synthesized peptides had no favorite solution conformation, suggesting that both the amino acid sequence and the conformation in the intact protein are necessary for antiviral action.

SARS-CoV-2 virus activates the human cell ACE2 receptor, triggering a series of deleterious events. In COVID19, renin-angiotensin is upregulated and the pathway is overexpressed and a progressive cytokine storm is always observed. In all these pathogenic processes, ovotransferrin could play a modifier function to enhance the condition. Ovotransferrin can be beneficial to the antioxidant effects of Angiotensin II by inhibiting NF-kB. Ovotransferrin is an immunomodulator that downregulates pro-inflammatory cytokines including TNF-α, IL-6, and IL-1 that could extinguish the cytokine storm. Ovotransferrin can also serve as a protective agent against COVID19 cardiotoxicity due to disruption in the ACE2-mediated signaling pathway, cytokine storm, pulmonary dysfunction, and side effects of medications.

In patients with coronavirus infection, provided ovotransferrin's possible protective effects, it is suggested as a supportive and therapeutic alternative.

Given this background, in the light of the current COVID-19 emergency, it is the intention of the investigators to conduct a prospective, randomized, open-label, controlled study in the cohort of hospitalized patients with covid-19 pneumonia, administering 200 mg of ovotransferrin orally in addition to the standard of care therapy (SOC).

The investigators hypothesize that the use of ovotransferrin will be associated with an earlier improvement of clinical and humoral parameters after 10 days of ovotransferrin treatment when compared to the group of patients provided with standard care.

• Study Type: Interventional
• Enrollment (Anticipated): 100
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Antonio Cascio, MD, PhD; Phone Number: +393389912198; Email: antonio.cascio03@unipa.it

Study Locations

• Italy
  • Palermo, Italy, 90127
    • Azienda Ospedaliera Universitaria Policlinico Paolo Giaccone
    • Contact: Antonio Cascio, MD; Phone Number: +393389912198; Email: antonio.cascio03@unipa.it

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: All

Description

Inclusion Criteria:

• Positive swab test of SARS-CoV-2
• Pneumonia related to SARS-CoV-2
• Signature of informed consent
• Absence of allergies to egg proteins

Exclusion Criteria:

• Unsigned informed consent
• Negative swab test of SARS-CoV-2
• Allergy to egg proteins
• Pregnancy and breastfeeding
• Mechanical ventilation

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
No Intervention: Standard of care
Experimental: Ovotransferrin; Dietary Supplement: Ovotransferrin	Dietary supplement: Ovotransferrin; Administering 200 mg of ovotransferrin orally in addition to the standard of care therapy for 10 days

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
In-hospital mortality	Change of hospital mortality	48hours

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Hospital stay	Length of hospital stay	up to 24 weeks
Duration of positive PCR swab	Time length of negativization of PCR molecular swab	5 days
C reactive protein (CRP) levels	Reduction of CRP levels > 50% in comparison with CRP levels at the admission, within 48 hours after the administration	48 hours
IL-6 levels	Reduction of IL-6 levels > 50% in comparison with IL-6 at the admission, within 48 hours after the administration	48 hours
D-dimer levels	Reduction of D-dimer levels > 50% in comparison with D-dimer at the admission, within 48 hours after the administration	48 hours

Collaborators and Investigators

• Sponsor: Azienda Ospedaliera Universitaria Policlinico Paolo Giaccone Palermo
• Investigators: Principal Investigator: Antonio Cascio, MD, PhD, AOU Policlinico "P. Giaccone" - University of Palermo, Italy

Publications and helpful links

General Publications

• Abeyrathne EDNS, Huang X, Ahn DU. Antioxidant, angiotensin-converting enzyme inhibitory activity and other functional properties of egg white proteins and their derived peptides - A review. Poult Sci. 2018 Apr 1;97(4):1462-1468. doi: 10.3382/ps/pex399. Review.
• Benedé S, Molina E. Chicken Egg Proteins and Derived Peptides with Antioxidant Properties. Foods. 2020 Jun 3;9(6). pii: E735. doi: 10.3390/foods9060735. Review.
• Chen S, Jiang H, Peng H, Wu X, Fang J. The Utility of Ovotransferrin and Ovotransferrin-Derived Peptides as Possible Candidates in the Clinical Treatment of Cardiovascular Diseases. Oxid Med Cell Longev. 2017;2017:6504518. doi: 10.1155/2017/6504518. Epub 2017 Mar 13. Review. Erratum in: Oxid Med Cell Longev. 2018 Dec 13;2018:1897257.
• Giansanti F, Massucci MT, Giardi MF, Nozza F, Pulsinelli E, Nicolini C, Botti D, Antonini G. Antiviral activity of ovotransferrin derived peptides. Biochem Biophys Res Commun. 2005 May 27;331(1):69-73.

Study record dates

Study Major Dates

• Study Start (Anticipated): December 1, 2020
• Primary Completion (Anticipated): February 28, 2021
• Study Completion (Anticipated): March 30, 2021

Study Registration Dates

• First Submitted: November 22, 2020
• First Submitted That Met QC Criteria: November 23, 2020
• First Posted (Actual): November 24, 2020

Study Record Updates

• Last Update Posted (Actual): November 24, 2020
• Last Update Submitted That Met QC Criteria: November 23, 2020
• Last Verified: November 1, 2020

More Information

Terms related to this study

• Keywords: Ovotransferrin
• Additional Relevant MeSH Terms: Coronavirus Infections; Coronaviridae Infections; Nidovirales Infections; RNA Virus Infections; Virus Diseases; Infections; Respiratory Tract Infections; Respiratory Tract Diseases; Pneumonia, Viral; Pneumonia; Lung Diseases; COVID-19
• Other Study ID Numbers: 20201116

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: No

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No