International Conference on Contemporary Science and Clinical Pharmacy, Indonesia-2023 | 11 STUDY OF ACTIVE ISOLATED COMPOUNDS FROM SUNGKAI LEAF (PERONEMA CANESCENS JACK) AS IMMUNOSTIMULANT FROM EXPOSURE OF THE SARS-COV-2 VIRUS ANTIGEN TO NATURAL KILLER CELLS Original Article DWISARI DILLASAMOLA 1* , YUFRI ALDI 2 , FATMA SRI WAHYUNI 3 , RAUZA SUKMA RITA 4 , YOHANNES ALEN 5 , DACHRIYANUS 6 1,2,3,5 Faculty of Pharmacy, Universitas Andalas, Pauh, Padang-25163, Indonesia. 4 Faculty of Medicine, Universitas Andalas, Padang-25163, Indonesia * Corresponding author: Dwisari Dillasamola; * Email: dwisaridillasamola@phar.unand.ac.id Received: 11 Oct 2023, Revised and Accepted: 23 Nov 2023 ABSTRACT Objective: SARS-CoV-2 is an illness that attacks the respiratory tract's alveoli lining cells. One approach to tackling it is to strengthen the immune system of the body, which has an organ specifically dedicated to defending against diseases. Among the key components of the body's immune system that defend against the SARS-CoV-2 virus are Natural Killer (NK) cells. Sungkai (Peronema canescens Jack) is a plant commonly used by communities to boost their immunity. The aim of this study was to examine the activity and characterization results of Sungkai leaves on Natural Killer cells. Method: Sungkai powder was extracted using a 70% ethanol solvent and evaporated using a rotary evaporator. It was then fractionated using n- Hexane and evaporated using a rotary evaporator to obtain a thick fraction. The thick fraction was then chromatographed, resulting in two isolated compounds that were tested in vivo for their immunostimulatory activity using mice as test animals with ethical approval (405/UN.16.2/KEP- FK/2021). Results: The study consisted of five groups, including a normal group and four other groups of white mice that had been exposed to the SARS-CoV-2 virus antigen Moderna vaccine 0.0013 ml. Conclusion: The results showed that stigmasterol was more effective than Bis(2-ethylhexyl) phthalate in increasing Natural Killer cells' immunostimulant activity. Keywords: SARS-CoV-2, Sungkai (Peronema canescens Jack), Natural Killer (NK), pthalate, Stigmasterol, Immunostimulant © 2024 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/) DOI: https://dx.doi.org/10.22159/ijap.2024.v16s1.02 Journal homepage: https://innovareacademics.in/journals/index.php/ijap INTRODUCTION The mysterious Pneumonia case that was first reported in Wuhan, Hubei Province in early 2020 has shocked the world. The rapid spread of the disease has affected more than 190 countries worldwide with an unclear etiology. The case has spread to various provinces in China, Thailand, Japan, and South Korea, and there were 44 new cases recorded from December 31, 2019, to January 3, 2020 [1]. The disease was named Coronavirus Disease 2019 (COVID-19) caused by the Severe Acute Respiratory Syndrome Corona Virus 2 (SARS-CoV-2). The virus has spread worldwide and infected more than 214 million people as of August 2021. In the last two decades, there have been three coronavirus outbreaks, namely Severe Acute Respiratory Syndrome (SARS), Middle East Respiratory Syndrome (MERS), and COVID-19 [2]. In humans, SARS-CoV-2 mainly infects cells in the respiratory tract that line the alveoli. SARS-CoV-2 binds to receptors and enters the cell. The body contains an immune system that protects against diseases. The key component of the immune system is white blood cells that can spread throughout the body through blood vessels. A crucial component of the immune system in protecting the body against SARS-CoV-2 infection is natural killer cells [3]. The body contains an immune system that protects against diseases. The key to the immune system is white blood cells, which can spread throughout the body through blood vessels. The body's defense mechanisms can be enhanced with certain compounds that act as immunostimulants. Immunostimulants are generally defined as compounds that can enhance the body's defense mechanisms, both specific and non-specific, as well as cellular and humoral defense mechanisms. Therefore, the presence of chemical compounds that can enhance the activity of the immune system, which can be obtained from plants, is important. One of the important components of the immune system in protecting the body against SARS-CoV-2 virus infection is Natural Killer (NK) cells [4]. These cells protect the body against SARS-CoV-2 virus infection by producing a group of cytokines, such as interferon, granzyme, and perforin, as well as proinflammatory cytokines known as the cytokine storm. In addition, the body also protects itself through phagocytosis, regardless of the differences among foreign substances. The body's defense mechanisms can be enhanced with specific compounds through an immunomodulatory approach [5]. Natural Killer (NK) cells play a central role in antiviral immunity. They can lyse virus-infected host cells by inducing apoptosis through several different pathways. Specifically, they can release various proteins such as perforin and granzyme through exocytosis, and together, these molecules can induce apoptosis of target cells. Second, NK cells can kill targets by expressing FAS ligand (FasL) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) as killer molecules and then initiate extrinsic apoptosis signaling pathways [6]. In addition, NK cells are capable of secreting pro- inflammatory cytokines, such as IFNy, TNF-a, GM-CSF, and chemokines. NK cells exhibit functional plasticity in the context of disease development by displaying pro-inflammatory or anti- inflammatory properties. Due to the reduced frequency of NK cells found in SARS-Cov-2 patients and the well-known antiviral role of NK cells, it is very intriguing to speculate that the recovery of NK cells and their function would be a good solution for SARS-Cov-2 patients [7]. The literature review on the extract of the Peronema canescens Jack plant found that a dose of 0.5625 mg/kgBW of Peronema canescens Jack increased leukocyte counts by 36% [8]. The ethanol extract from Peronema canescens Jack leaves had in vivo antiplasmodium activity, with an ED50 value of 102.88 mg/kgBW. N-hexane extract from Peronema canescens Jack leaves inhibited the growth of all test International Journal of Applied Pharmaceutics ISSN- 0975-7058 Vol 16, Special Issue 1, 2024