Phase I, Safety and Immunogenicity of Razi SARS-CoV-2 recombinant Spike protein vaccine (Razi Cov Pars)

INTERVENTION: Intervention 1: Intervention group 1: Vaccine at 5 µg/200µl dose; Participants in this group will receive two doses (IM) of RAZI recombinant spike protein vaccine 21 days apart followed by a 10 µg/200µl nasal spray 51 days after the first dose (day 0). Intervention 2: Intervention group 2: Vaccine at 10 µg/200µl dose; Participants in this group will receive two doses (IM) of RAZI recombinant spike protein vaccine 21 days apart followed by a 10 µg/200µl nasal spray 51 days after the first dose (day 0). Intervention 3: Intervention group 3: Vaccine at 20 µg/200µl dose; Participants in this group will receive two doses (IM) of RAZI recombinant spike protein vaccine 21 days apart followed by a 10 µg/200µl nasal spray 51 days after the first dose (day 0). Intervention 4: Control group: Adjuvant; Participants in this group will receive two doses (IM) of Adjuvant by 50% v/v concentation produced in RAZI institute 21 days apart followed by another dose in the form of nasal spray at day 51 (counted from day 0). CONDITION: SARS‐CoV‐2. ; COVID‐19 U07.1 SECONDARY OUTCOME: Neutralizing antibody activity: Neutralizing antibody titers will be measured on day zero and day 35 (2 weeks after the second dose) in all participants. Measurements in other times will only be performed on 20% of participants. The following tasks will be performed during the conduct of this test. 1‐ In vitro assessment of inhibitory effect of antibody on the binding of Spike antigen with human ACE2 receptor and 2‐ Assessing VNT titer. Timepoint: Humoral immunity will be assessed based on the neutralizing antibody titers on days 0, 35, 65, and months 5, 8, and 12, and comparisons will be done with day 0. Method of measurement: Virus Nutralization Test (VNT). Number and percentage of Covid‐19 disease occurrence two weeks after second vaccine dose. Timepoint: ?Any time between the 14 days after second vaccine dose and the end of study. Method of measurement: Diagnosis of Covid‐19 disease will be based on Iran's Ministry of Health's guideline and a positive positive PCR test. Number and percentage of Medically Attended Adverse Events (MAAEs). Timepoint: Monthly until sixth month after last vaccine dose. Method of measurement: These events will be collected monthly from the participants through face to face or telephone contacts. In case of medically attended events identification in the participants, more information about the event will be collected and discussed at the DSMB meeting. Number and percentage of Severe Adverse event (SAEs). Timepoint: Monthly until sixth month after last vaccine dose. Method of measurement: These events will be collected monthly from the participants through face to face or telephone contacts. In case of Severe Adverse event identification in the participants, more information about the event will be collected and discussed at the DSMB meeting. Number and percentage of Suspected Unexpected Serious Adverse Reaction(SUSAR ). Timepoint: Monthly until sixth month after last vaccine dose. Method of measurement: These events will be collected monthly from the participants through face to face or telephone contacts. In case of Suspected Unexpected Serious Adverse Reaction identification in the participants, more information about the event will be collected and discussed at the DSMB meeting. Serum levels of specific IgG antibodies against S, S1, S2, NTC, RBD components of SARS‐CoV‐2 spike protein antigen(s). "serum antibody level" will be assessed using GMT geometric mean, as well as "serum fold rise" and "seroconversion rate " and will be compared between the two groups. Seroconversion rate is defined as the the proportion of the individuals whose serum IgG levels have two fold or more using the ELISA method. Changes in these factors as well as showing no response against N antigen will be explored. Timepoint: Days zero, 7, 14, 21, 28, 35, 51, 58, 65 and months 5, 8 and 12. Method of measurement: Will be measured using ELISA method. The cell‐mediated immunity will be evaluated by counting the number of CD3, CD4 and CD8 cells and joint calculation of CD3 and CD4 and CD3 and CD8 . IFN‐?, TNF‐a, and interleukins 2, 4, 6, and 17 will also be measured. Evaluation of cell mediated immunity will be performed only in 20% of participants in each group. Cell mediated immunity will be measured in all participants on day 35 (2 weeks after the second dose). Summary of the measures performed in this section are as follows: 1‐ Assessment of CD4 to CD8 cell proportions after stimulation of PBMC (Peripheral Blood Mononuclear Cells) by inactivated virus and recombinant spike protein using flow cytometry 2‐ Assessment of specific proliferation of PBMC cells stimulated by inactivated virus and recombinant spike protein using flow cytometry 3 ‐ Assesment of TH1 and TH2 specific cellular immunity after PBMC stimulation in vaccinated individuals with recombinant spike protein to determine the levels of interferon‐gamma, interleukin‐4, tumor necrosis factor‐alpha and interleukin 6 using Eli spot and ELISA kit. Timepoint: Cell mediated immunity will be assessed on days 0, 35 and 65 and months 5, 8 and 12 and comparison will be made between day 0 and other time points. Method of measurement: Immunologic lab tests. INCLUSION CRITERIA: Having Iranian citizenship; Diploma or higher degree; Adults aged 18 ‐ 55 years; Body mass index 17 to 35 kg/m2; Having good health based on clinical and laboratory criteria; Having sublingual temperature less than or equal to 37.2 ° C in the morning based on mercury thermometer; Negative IgG and IgM antibody titers for COVID‐19 S antigen; Negative RT‐PCR test for COVID‐19; Negative IgG ELISA for HIV; Having heart rate between 60 and 100; Signed the informed consent form; The participant agrees to reduce the risk of developing of COVID‐19; For females of childbearing age 18 to 49 years: not being pregnant based on the first day of the last menstrual period; For females of childbearing age 18 to 49 years: negative pregnancy test based on bHCG on the day of screening and the day of vaccination; For females of childbearing age 18 to 49 years: use at least one effective method of contraception (condoms, oral contraceptive pills PRIMARY OUTCOME: Abnormal vital signs and anaphylactic reactions before and immediately after vaccination: number and percentages of participants who develop abnormal vital signs within three hours of receiving the vaccine at each doses will be recorded. Abnormal vital signs include temperature, respiratory rate, heart rate, systolic and diastolic blood pressure. Anaphylaxis is defined as an immediate systemic hypersensitivity simultaneously involving two systems. Anaphylactic reactions include: erythema, pruritus, urticaria and angioedema, bronchospasm, laryngeal edema, dizziness, hypotension, nausea, shortness of breath, wheezing, arrhythmia, cyanosis, vomiting, diarrhea, abdominal pain and will be checked up to three hours after each vaccination. Timepoint: Before vaccination and hourly for three hours after vaccination at each dose. Method of measurement: Clinical examination. The number and percentage of ocal adverse events within the first week post‐vaccination (including pain, tenderness, erythema / redness, swelling and stiffness, itching) that will be assessed based on the severity score, duration and peak intensity. Timepoint: Seven days after 1st and 2nd vaccination(Days 0‐7 and 21‐27)daily assessment. Method of measurement: Daily symptom registration cards will be given to patients at the time of vaccination and they will be asked to bring them with themselves on the next visit. These patients will be contacted daily during these seven days and the study staff will ensure that the cards are completed. The number and percentage of people who shows abnormal laboratory findings, including biochemistry, hematology, and urine tests. These tests include: Hemoglobin, WBC, Lymphocytes cell, Neutrophils, Eosinophils, Platelets, ESR, CRP, LDH, Sodium, Potassium, HbA1c, BUN, Creatinine, Calcium, Alkaline phosphatase, ALT, AST, Bilirubin (total), Uric Acid, U/A, Urine protein, Urine glucose, Urine RBC. Timepoint: 7 Days after each vaccine dose (Days 7, 28, 58). Method of measurement: Each test will be performed using the appropriate kit. The number and percentage of systemic adverse event within the first week post‐vaccination (including nausea and vomiting, diarrhea, headache, fatigue, muscle pain) that will be assessed based on the severity score, duration and peak intensity. Timepoint: Seven days after each vaccination step (Days 0‐7 and 21‐27 and 51‐57) daily assessment. Method of measurement: Daily symptom registration cards will be given to patients at the time of vaccination and they will be asked to bring them with themselves on the next visit. These patients will be contacted daily during these seven days and the study staff will ensure that the cards are completed.