Study protocol for a randomized, open-label, non-controlled Phase I/II Study to assess safety and immunogenicity of twice or three times dosing of intramuscular COVID-19 DNA vaccine in healthy adults

Abstract

There is currently an outbreak of respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and many vaccines have been rapidly developed using different technologies. We have challenged to develop DNA vaccines for SARS-CoV-2. After the basic and preclinical experiments, AnGes, Inc. conducted a randomized, open-label, non-controlled Phase I/II Study to assess safety and immunogenicity of intramuscular DNA vaccine (AG0302-COVID19) in Osaka University Hospital. Thirty healthy volunteers, male or female, aged 20–65, will be randomized to one of the following three groups, 1) 2.0 mg of AG0302-COVID19 twice at 2-week intervals, 2) 2.0 mg of AG0302-COVID19 twice at 4-week intervals, 3) 2.0 mg of AG0302-COVID19 three times at 2-week intervals. This study will assess the safety and immunogenicity of AG0302-COVID19. The primary endpoint about the safety and tolerability has been assessed by the incidence of treatment emergent adverse events from week 1 through week 9, and frequency and severity of each adverse event, including solicited local and systemic adverse events through 8 weeks after the first vaccination. The immunogenicity has been also assessed by the change in geometric mean titer (GMT) of serum anti-SARS-CoV-2 Spike glycoprotein specific antibody. This study has been registered at the ClinicalTrials.gov.as NCT04527081.

Highlights

● We conducted a randomized, open-label, non-controlled Phase I/II Study to assess safety and immunogenicity of intramuscular DNA vaccine (AG0302-COVID19) in Osaka University Hospital.

● Thirty healthy volunteers, male or female, aged 20–65, will be randomized to one of the following three groups, 2.0 mg of AG0302-COVID19, 1) twice at 2-week intervals, 2) twice at 4-week intervals, 3) three times at 2-week intervals.

● The primary endpoint is the safety and tolerability. The immunogenicity has been also assessed by the change in geometric mean titer (GMT) of serum anti-SARS-CoV-2 Spike glycoprotein specific antibody.

Introduction

Many pharmaceutical companies are developing vaccines against SARS-CoV-2, including vaccines based on inactivated target virus or viral proteins, adenovirus vectors and RNA/DNA from SARS-CoV-2 [1,2,3,4,5,6]. The major target antigen is the spike glycoprotein of SARS-CoV-2, which is essential for virus entry into cells [7]. We have challenged to develop DNA vaccines for SARS-CoV-2, that can be easily developed to use only genetic material, not viruses, and potentially activate cellular immunity as well as humoral immunity. In the history of DNA vaccine, the first clinical trial has been started in 1990’s, and a lot of clinical trials that focus on DNA vaccination are registered. Overall, no severe adverse events related to DNA vaccine have been reported in the safety evaluation. A few DNA-based vaccines were approved for veterinary use, including a vaccine against West Nile Virus in horses [8] and canine melanoma [9]. In terms of plasmid DNA vaccines for COVID-19 in humans, INOVIO developed a DNA vaccine which are intradermally delivered by electroporation [10], and Zydus in India also developed a plasmid DNA vaccine which are intradermally delivered by Bioinjectors. Whereas, we will develop a plasmid DNA vaccine which are delivered into muscle by a simple injection.

Protocol Digest for the Study

Objective

This study will assess the safety and immunogenicity of AG0302-COVID19 in healthy adult volunteers.

Study setting

This study is a randomized, open-label, non-controlled Phase I/II Study to assess safety and immunogenicity of twice or three times dosing of intramuscular AG0302-COVID19 (2 mg) in healthy adults.

Endpoints

1. Primary endpoint

Safety and tolerability: Incidence of Treatment-Emergent AE (Adverse Events (Week 1 through Week 9), and frequency and severity of each adverse event, including solicited local and systemic AEs 8 weeks after the first vaccination.

Immunogenicity (Weeks 3, 5, 7, 9): Change in Geometric mean titer (GMT) of serum anti-SARS-CoV-2 Spike (S) glycoprotein-specific antibody.

2. Secondary endpoint

Change in GMT of anti-S specific antibody, GMT of anti-S receptor binding domain-specific antibody, GMT of anti-SARS-CoV-2 B cell epitope antibody, IgG subclasses (IgG1 and IgG2) of anti-S-specific antibody, the neutralizing activity against pseudovirus of SARS-CoV-2, binding inhibition of S and ACE2, IFN-γ production against S in peripheral blood mononuclear cells, adverse events.

Eligibility criteria

1. Inclusion

1) Subjects who have obtained written consent voluntarily to participate in this clinical trial.

2) Subjects whose age at the time of obtaining consent is 20 years to 65 years.

3) Subjects who are negative for SARS-CoV-2 by PCR test.

4) Subjects who are negative for both SARS-CoV-2 IgM antibody and SARS-CoV-2 IgG antibody by antibody test.

2. Exclusion

1) Subjects with symptoms of suspected COVID-19 infection (respiratory symptoms, headache, malaise, olfactory disorders, taste disorders, etc.).

2) Subjects with a history of COVID-19 (hearing from subjects).

3) Subjects who have participated in unapproved vaccine clinical trials.

4) Subjects with axillary temperature of 37.0 degree or higher.

5) Subjects who have a history of anaphylaxis.

6) Subjects who have a current or history of serious renal, cardiovascular, respiratory, liver, kidney, gastrointestinal, and neuropsychiatric diseases.

7) Subjects with a history of convulsion or epilepsy.

8) Subjects with a history of diagnosis of immunodeficiency.

9) Subjects who have a close relative (within 3rd degree) of congenital immunodeficiency.

10) Subjects who have current bronchial asthma.

11) Subjects who had a fever of 39.0°C or higher within 2 days after the last vaccination, and those who suspected allergy such as a systemic rash.

12) Females who wish to become pregnant from the date of study registration to 12 weeks after the first inoculation of the investigational drug, and pregnant females who are breast-feeding. In addition, females who may become pregnant and their male sexual partners should use appropriate contraceptives (pill), condoms, vasectomy, tubal ligation, diaphragm, intrauterine devices, spermicides, intrauterine hormone-releasing system, etc. from the study entry date until 12 weeks after the first vaccination.

13) Subjects who have participated in clinical trials of other unapproved drugs and received the investigational drug within 4 weeks before the start of this clinical trial (starting from vaccination day).

14) Subjects who have been received a live vaccine, inactivated vaccine, or toxoid within 4 weeks before the start of this clinical trial (starting from vaccination day).

15) Subjects who have been administered with drugs that affect the immune system (excluding external preparations) such as immunomodulators (DMARDs, etc.), immunosuppressants, biologics, etc. within 4 weeks before vaccination.

16) Subjects who received blood transfusion or gamma globulin therapy within 12 weeks before vaccination, or high-dose gamma globulin therapy (200 mg/kg or more) within 24 weeks before vaccination.

17) Subjects who have a history of overseas travel within 4 weeks before the start of the clinical trial (starting from vaccination day).

18) Subjects who are unable to comply with the clinical trial protocol and follow up (for mental, family, social or geographical reasons).

19) Subjects who are judged to be ineligible for this clinical trial by the investigator.

Treatment

Thirty healthy volunteers, male or female, aged 20–65, will be randomized to one of the following three groups.

1. 2.0 mg of AG0302-COVID19 twice at 2-week intervals.

2. 2.0 mg of AG0302-COVID19 twice at 4-week intervals.

3. 2.0 mg of AG0302-COVID19 three times at 2-week intervals.

Clinical trials registry

This study has been registered at the ClinicalTrials.gov.as NCT04527081. This study initiated on August 31, 2020 and continued until September 30, 2021.

Conflict of Interest

The Department of Health Development and Medicine is an endowed department supported by AnGes, Daicel, and FunPep. The Department of Geriatric Medicine and Health Development and Medicine has clinical trial and basic experiment agreement with Anges. The Department of Clinical Gene Therapy is financially supported by Novartis, AnGes, Shionogi, Boeringher, Fancl, Saisei Mirai Clinics, Rohto and Funpep. R.M. is a stockholder of AnGes.

Acknowledgment

This study has been supported by a Project Promoting Support for Drug Discovery grant (JP20nk0101602) from the Japan Agency for Medical Research and Development.

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