Development of SARS-CoV-2 Therapeutics from APRG64

다국어 초록 (Multilingual Abstract)

The coronavirus is a virus that belongs to the family Coronavirusicae, commonly found in various mammals, including humans and birds. Moreover, it is known to cause both respiratory and digestive infections depending on the nature of the virus and its host. The main symptoms of COVID-19 are respiratory symptoms, such as fever, cough, sore throat, and difficulty breathing, while depending on the patient, it can also appear in other various forms of ailment, such as headache, muscle pain, hemoptysis, chills, chest pain, diarrhea, decreased taste and smell, and loss of lung function. Healthy adults are likely to recover over time, but it can be fatal to people with low immune functions, such as the elderly and those with underlying diseases. Notably, COVID-19 is still spreading rapidly worldwide, and there is no suitable treatment due to the appearance of mutant viruses. Therefore, there is a need to develop a therapeutic agent that can treat COVID-19.
APRG64, which was developed in this study, is an extract mixture of Agrimonia pilosa L. and Galla rhois. It has been confirmed through previous studies that its effect on inhibiting COVID-19 is more than 50 times that of remdesivir. Based on these results, a study was conducted to develop APRG64 as a COVID-19 therapeutic.
APRG64 is a green, round, film-coated tablet that has been completely produced. This study established the specifications for Agrimonia pilosa L. dried extract, Galla rhois dried extract, and APRG64. Moreover, APRG64 was confirmed to be a safe substance that does not affect genotoxicity and reproductive toxicity through preclinical safety tests. The LD50 and NOAEL were confirmed in SD rats and beagle dogs through oral toxicity tests (SD rats: 2,000 mg/kg/day; beagle dog: 500 mg/kg/day). As a result of confirming the antiviral activity of APRG64 using the Syrian hamster, the lung virus was significantly reduced when 100 mg/kg was administered. It was confirmed that the mortality rate was 0% when 50 mg/kg or more was administered. The molecular docking test confirmed that the active ingredient of APRG64 had an antiviral effect by binding to the spike protein RBD in the coronavirus and inhibiting the ability to bind to ACE2. To confirm the safety and tolerability of APRG64, a phase 1 clinical trial was conducted on 40 healthy adult males. As a result, no severe adverse reactions occurred, and safety and tolerability were confirmed at 400 mg, 800 mg, and 1200 mg.
These results confirmed that APRG has excellent efficacy as a candidate substance for treating COVID-19.

목차 (Table of Contents)

• Abstract i
• Contents iii
• Abbreviations vi
• List of Tables vii
• List of Figures ix
• 1. Introduction 1
• 2. Materials and Methods 4
• 2.1. Establishment of drug substance manufacturing process and specifications 4
• 2.1.1. Manufacturing drug substances 4
• 2.1.2. Test methods of dried extract for specifications 5
• 2.1.3. Stability test of drug substances 10
• 2.2. Establishment of the APRG64 manufacturing process and specifications 11
• 2.2.1. Manufacturing APRG 64 11
• 2.2.2. Test methods of APRG64 for specifications 13
• 2.3. Antiviral activity of APRG64 against SARS-CoV- 2 16
• 2.3.1. Antiviral activity against SARS-CoV-2 in an animal model 16
• 2.3.2. Antiviral activity against SARS-CoV-2 using molecular docking 18
• 2.3.3. Neutralization against SARS-CoV-2 spike RBD by APRG 64 21
• 2.4. Phase I clinical trial of APRG 64 22
• 2.4.1. Overall clinical trial design and description of the process 22
• 2.4.2. Study endpoint 23
• 2.4.3. Selection of subject 24
• 2.4.4. Investigational products 29
• 2.4.5. Efficacy and Safety Variables 31
• 3. Results 36
• 3.1. Specification of drug substances 36
• 3.1.1. Arimonia pilosa L. dried extract manufacturing and analysis 36
• 3.1.2. Galla rhois dried extract manufacturing and analysis 37
• 3.1.3. Stability test of Agrimonia pilosa L. dried extract (3 months) 39
• 3.1.4. Stability test of Galla rhois dried extract (3 months) 43
• 3.2. Specification of APRG 64 47
• 3.3. Antiviral activity results of APRG64 against SARS-CoV- 2 52
• 3.3.1. Results of the antiviral activity in an animal model 52
• 3.3.2. Result of antiviral activity using molecular docking 58
• 3.3.3. RBD binding inhibition of SARS-CoV by APRG 64 60
• 3.4. Result of phase I clinical trial of APRG 64 62
• 3.4.1. Extent of exposure 62
• 3.4.2. Adverse events (AEs) 62
• 3.4.3. Deaths, other serious adverse events 64
• 3.4.4. Other significant adverse events 64
• 3.4.5. Clinical laboratory results 65
• 3.4.6. Vital signs and other safety-related observations 65
• 3.4.7. Phase I clinical trial conclusion and discussion 66
• 4. Conclusion and Discussion 74
• Reference 76
• Abstract in Korean 80