Production of Protective Lozenges / Chewable Tablets Against SARS-CoV-2 Virus

Abstract

This invention relates to the production of a mixture (18) consisting of extracts of green tea, blueberry, pomegranate peel and Black Poplar propolis. Unexpectedly, after the mixture (18) showed efficacy against the SARS-CoV-2 virus, the mixture (18) was converted into a lozenge / chewable tablet form and its efficacy against the SARS-CoV-2 virus was demonstrated. The invention was developed in the form of a throat lozenge / chewable tablet to provide a protective effect around the mouth and throat, which is the first entry point of the SARS-CoV-2 virus. The efficacy of the mixture (18) against the SARS-CoV-2 virus was tested in vitro on VERO E6 cells. At the end of the 160-hour experiment, the mixture (18) neutralized the virus at a concentration of 110 μg / mL at a rate of 112% (61% cell viability) and at a concentration of 70 μg / mL at a rate of 35% (82% cell viability).

Description

TARIFF Production of Protective Lozenges / Chewable Tablets Against SARS-CoV-2 Virus The Technical Field to Which the Discovery Belongs This invention relates to the production and formulation of a mixture of green tea, blueberry, pomegranate peel, and black poplar propolis extracts, whose efficacy against the SARS-CoV-2 virus has been proven in vitro (on VERO E6 cells). The effectiveness of the formulated mixture against the SARS-CoV-2 virus has been demonstrated in vitro. Cytotoxicity tests have also been performed on the developed product. The reason for developing the product in lozenge / chewable tablet form is to provide a protective effect around the mouth and throat, which are the initial entry points for the SARS-CoV-2 virus. State of the Art Starting in China in 2019 and spreading throughout the world, Covid-19 (Coronavirus pandemic) has become an epidemic threatening humanity. SARS-CoV-2, the causative agent of Covid-19, is a contagious viral disease that causes serious respiratory tract infections in humans. SARS-CoV-2 is an enveloped virus that can infect humans and some animal species, carrying single-stranded RNA (ribonucleic acid) as its genetic material. This group of viruses mainly causes upper respiratory tract infections, showing cold and flu-like symptoms. Sore throat, dry cough, runny nose, weakness and fatigue are among the most common symptoms [5]. To date, the number of Covid-19 cases worldwide has exceeded 185 million, and more than 4 million deaths have occurred [1]. Covid-19 has significantly impacted the health systems of countries globally. In addition, it has caused problems in the fields of education and economy, and has greatly affected the food and tourism sectors. Furthermore, the isolation of society has limited the social lives of individuals, leading to the development of psychological problems. The rapid spread of Covid-19 among people and the high mortality rate it causes have created an urgent need for the development of new treatment methods for the disease. While there is currently no known effective method to treat Covid-19 worldwide, 19 different vaccines have been developed [1]. Clinical trials of potential antiviral drugs are ongoing. Clinical studies have shown that plant extracts are of great importance in the development of drugs for the treatment of viral diseases thanks to their rich bioactive and phytochemical compound content [6]. Plants synthesize secondary metabolites with a wide range of biological activity to grow and survive. Many of these metabolites are chemicals that form defense mechanisms in the plant against biotic stress (insects, parasites and microbial pathogens) and abiotic stress (low or high temperature, insufficient or excessive water, saline environment, heavy metals and UV light) [7]. For centuries, people have used plant extracts for the treatment and prevention of diseases. The therapeutic effects of various and numerous plant-derived compounds against SARS-CoV-2 are being investigated. Currently, there are dietary supplements used to strengthen the immune system. These supplements, available in tablet, capsule, and lozenge forms, contain various combinations of beta-glucan, elderberry extract, African geranium, turmeric extract, grape seed extract, green tea extract, and pomegranate peel extract, along with vitamins and minerals. Especially in throat infections, lozenges / chewable tablets, used to reduce pain and facilitate swallowing, stimulate salivary gland activity. This increases the secretion of lysozyme, a naturally occurring antimicrobial substance in the mouth, thus preventing the growth and colonization of microorganisms entering the body through the mouth and nose, including in newly developing throat infections. Furthermore, it is reported that SARS-CoV-2 and its variants penetrate cells by attaching to ACE-2 and TMPRSS2 receptors. The highest concentration of these receptors is found in the oral cavity and nasal mucosa. Therefore, lozenges / chewable tablets may play an important role in preventing viral attachment and penetration to these receptors in the oral mucosa. The existing patents and applications are summarized below. - Patent document number AU2020100565, titled “A composition to boost and enhance immune system and application thereof,” focuses on enhancing the immune system and, unlike the current invention, includes extracts of Terminalia ferdinandiana, Undaria pinnatifida, Curcuma longa, and Vitis vinfera. - Patent document number US20160095879, titled “Composition for enhancing immunity,” differs from the existing invention in that it focuses on the development of food supplements, natural preparations, or formulations for the regulation of the general immune system, not against SARS-CoV-19. - The Japanese patent document numbered JP2005314316A, titled "Anti-SARS Coronavirus Agent" against SARS-CoV-2, mentions the development of food products containing proanthocyanidin, catechin, and grape extract that act as coronavirus agents, differing from the current invention. - In the Turkish patent document numbered TR2021 / 01716, titled "New Options with Antiviral Effects Against SARS-CoV-2," it is mentioned that, unlike the existing invention, combinations of Vitis vinifera seed oil, Carvacrol, Thymol, and Gamma Aminobutyric Acid with potential antiviral effects against Covid-19 have been developed. - The Turkish patent document numbered TR2020 / 16536, titled "Effective Herbal Food Supplement Against Covid-19," mentions the development of fermented liquid extracts with antiviral activity against Covid-19, obtained from different parts of plants / plants other than the plants and bee product mentioned in the existing invention. - Patent document number US10758587B2, titled “Formulations for the Treatment of Disorders of the Mouth, Throat and Respiratory Tract,” contains extracts of Curcuma longa, Punica granatum, and Zingiber officinale. In vitro and clinical studies have proven that this mixture is effective in preventing upper respiratory tract diseases by inhibiting bacterial plaque formation, and in treating upper respiratory tract diseases by increasing lysozyme levels in saliva, along with exhibiting antiviral, antioxidant, and anti-inflammatory effects. The patent application includes extracts of Punica granatum, Camellia sinensis, Vaccinium myrtillus, and the bee product, Black Poplar propolis extract, and its efficacy against the SARS-CoV-2 virus has been proven in vitro. - Patent document number US9173912B2, titled “Cistus extract containing enriched secondary plant ingredients,” describes extraction recipes for extracts of rockrose (Cistus creticus L.) containing a minimum of 40% (m / m) polyphenols, distinct from the present invention. Furthermore, it is stated that an extract containing 60% polyphenols is effective against Clostridium perfringens, Vibrio cholerae, and influenza A H1N1 viruses. The invention is described as being applicable in drop, nasal spray, syrup, tablet, lozenge, and powder forms. - Patent No. US 2012 / 0244087 A1, titled "Oral compositions containing a combination of natural extracts and related methods," concerns the preparation of a mixture obtained from a combination of at least three of the following plants: Punica granatum, Myristica fragrans, Zingiber officinale, and Zizyphus joazeiro, and its use in the formulation of oral care products. The resulting oral care products are stated to exhibit antimicrobial, anti-inflammatory, and antioxidant properties, and are also effective in the treatment of xerostomia without the need for another antibacterial agent. The characteristic of the mixture is that it can be taken in the desired form for application to the oral surface, such as liquid, powder, gel, chewable tablet, drops, or film. - Patent document number US20090175971A1, titled “Methods of using composition comprising pomegranate extracts against the common cold,” states that, unlike the present invention, a mixture obtained from two or more extracts of the whole pomegranate or any component of the pomegranate can be used in the treatment of the common cold caused by adenovirus, rhinovirus, coronavirus, and human parainfluenza virus, or any number of their variations, as well as any other viral agents. The majority of existing applications are inventions that strengthen the immune system and exhibit antiviral properties against viruses. In addition, the current invention includes plant and bee product extracts, as well as products consisting of different combinations of plant liquid extracts and synthetic and natural molecules that have potential antiviral activity against the SARS-CoV-2 virus. Green tea (Camellia sinensis) is obtained by drying freshly harvested Camellia sinensis leaves under hot air or steam without undergoing oxidation and fermentation. The most important phenolic compounds found in green tea are catechins [Epicatechin (EC), Epicatechin Gallate (ECG), Epigallocatechin (EGC), Epigallocatechin]. [Gallate (EGCG)] [8]. Catechins have been observed to have an important place in the prevention and treatment of cardiovascular diseases, obesity, diabetes, oxidative and inflammatory diseases, bacterial and viral diseases, cancer and neurological diseases [9,10]. Polyphenols in green tea help reduce body inflammation and have antioxidant and antiviral properties against SARS-COV-2. In addition, phenolic compounds in green tea also have antiviral properties against SARS-COV-2. Pomegranate (Punica granatum L.), which is widely cultivated in the Mediterranean region, is very rich in polyphenols such as ellagic tannins (ETs), gallic acid (GA), ellagic acid (EA), punicalin and anthocyanins

[11] . 40% of the pomegranate fruit is made up of peel. Pomegranate peel is very rich in dietary fiber and polyphenols and also has a much higher antioxidant capacity than the pulp

[12] . Studies have shown that pomegranate peel has antioxidant, antibacterial [13, 14], antimicrobial

[15] , antiviral [16, 17] effects and anti-inflammatory activity. ETs, GA, EA, and punicalin found in pomegranate peel have immune system strengthening and antiviral properties against SARS-CoV-2. Blueberry (Vaccinium myrtillus) is a plant species that grows in the temperate climate zone, and its fruit belongs to the group of berry fruits. In Türkiye, wild forms grow mainly in the Black Sea region, as well as in the Marmara Region and Eastern Anatolia flora. Blueberry is quite rich in phenolic compounds, especially anthocyanins (Malvidin, Delphinidin and Kuromarin) [18,19]. Thanks to the phenolic compounds and anthocyanins (Malvidin, Delphidin, Kuromarin etc.) it contains, blueberry has antiviral properties against SARS-COV-2. Propolis (Propolire) is a resinous material produced by honey bees to protect their hives and is widely used to strengthen the immune system. Containing phenolic compounds (Caffeic Acid Phenethyl Ester (CAFE), caffeic acid, p-coumaric acid, cinnamic acid, etc. Hydroxycinnamic acid derivatives) and flavonoids (galangin, chrysin, quercetin, pinosembrin, pinobanksin), propolis has immune-boosting, anti-inflammatory and antiviral properties against SARS-CoV-2

[20] . Among the effective components in the phenolic content of Black Poplar (Populus nigra) propolis, caffeic acid phenethyl ester (CAFE) is reported to have a broad spectrum of pharmacological activity in scientific studies. It has been observed that KAFE is not among the other three types of propolis (trembling aspen type, 3-methyl quercetin type, and a type with an unknown content) that have been identified in our country.In this respect, Black Poplar propolis is extremely important in terms of providing the biological benefits attributed to propolis. In addition, it is reported that pinosembrin, pinobanksin, galangin and chrysanthemum found in Black Poplar propolis are phenolic components responsible for the antimicrobial effect

[21] . Black Poplar propolis has immune-boosting, anti-inflammatory and antiviral properties against SARS-CoV-2 through the phenolic substances it contains; CAFFE, Caffeic acid, p-coumaric acid, t-cinnamic acid and hydroxycinnamic acid derivatives) and flavonoids (galangin, chrysanthemum, quercetin, pinosembrin, pinobanksin). The present invention relates to the formulation of extracts rich in phenolic compounds and possessing antiviral (anti-SARS-CoV-2) properties in specific ratios by converting them into powder form. Drying and powdering the bioactive-rich mixture is important for preventing spoilage, increasing shelf life, and facilitating ease of use. Currently, the knowledge available in the state of the art focuses on strengthening the immune system and exhibiting antiviral properties against viruses. In addition, the present invention includes plant and bee product extracts, as well as products consisting of various combinations of plant liquid extracts and synthetic and natural molecules with potential antiviral activity against the SARS-CoV-2 virus. Furthermore, unlike the prior art, the present invention includes bee products in addition to plant extracts. The black poplar (Populus nigra) propolis, a bee product, adds value to this invention due to its caffeic acid phenethyl ester (CAF) and rich polyphenol content, demonstrating its effectiveness against SARS-CoV-2 and other pathogens. The efficacy of black poplar propolis against SARS-CoV-2 has been demonstrated for the first time in this invention. Black poplar propolis, a bee product, has been added to broaden the efficacy profile of the invention. The present invention relates to a dietary supplement with antiviral efficacy against SARS-CoV-2, proven in vitro, which also strengthens the immune system, reduces the amount of virus transmitted through the oral and nasal tract [112% neutralizing activity (61% cell viability) at a concentration of 110 µg / ml], is easy to use, and has low toxicity. Furthermore, unlike other inventions, this invention includes bee products in addition to plant extracts. The Technical Problem That the Invention Aims to Solve While there is currently no known effective method to treat COVID-19 caused by SARS-CoV-2, which caused a pandemic worldwide, 19 different vaccines have been developed so far [1]. However, the development of suitable treatment methods is an urgent need. This invention relates to a mixture of extracts of green tea, blueberry, pomegranate peel, and black poplar propolis that targets host proteins, inhibits viral infection, and reduces the activity of the SARS-CoV-2 virus transmitted through the upper respiratory tract. The invention is developed in the form of a throat lozenge / chewable tablet to provide a protective effect around the mouth and throat, which are the initial entry points for the SARS-CoV-2 virus. The efficacy of the mixture against the SARS-CoV-2 virus was tested in vitro on VERO E6 cells. At the end of the 160-hour experiment, the mixture neutralized the virus at a concentration of 110 pg / mL at a rate of 112% (61% cell viability) and at a concentration of 70 pg / mL at a rate of 35% (82% cell viability). Plant-based substances are largely metabolized under varying pH and enzyme conditions in the digestive system. Therefore, it is unlikely that the results of in-vitro studies will systematically demonstrate the same efficacy throughout the body. However, when applied locally in the mouth (as lozenges / chewable tablets), it is possible to achieve the exact same efficacy as in-vitro results because they do not enter the digestive system. Thanks to the lozenge / chewable tablet form of this invention, the active ingredients exhibit local efficacy without being metabolized in the digestive system. Given that the oral cavity is considered the primary entry point for SARS-CoV-2 and other microorganisms, such a lozenge is expected to play an important role in preventing infections. Explanation of the Figures Figure 1. Obtaining a mixture with antiviral activity. Figure 2. Graph showing the neutralization efficacy of the mixture against SARS-CoV-2 virus on VERO E6 cells obtained from the xCELLigence RTCA MP device. Explanation of References in Figures A: Green Tea Extraction 1: 100% Ethanol 2: Temperature (A) 3.1: Filtering B: Filtration Process 1: 100% Ethanol 4.1: Filtrate C: Drying Process 5.1: Temperature (°C) 6.1: Green Tea Extract D: Blueberry Extraction 7: 50% Ethanol 8: Temperature (D) 3.2: Filtering B: Filtration Process 1: 100% Ethanol 4.2: Filtrate C: Drying Process 5.2: Temperature (°C) 6.2: Blueberry Juice Extract E: Lyophilization Process 9: Temperature (E) 10: Maltodextrin 11: Blueberry Extract F: Pomegranate Peel Extraction 12: 30% Ethanol 13: Temperature (F) 3.3: Filtering B: Filtration Process 1: 100% Ethanol 4.3: Filtrate C: Drying Process 5.2: Temperature (°C) 6.3: Pomegranate Peel Water Extract E: Lyophilization Process 9: Temperature (E) 10: Maltodextrin 14: Pomegranate Peel Extract G: Black Poplar Propolis Extraction 15: 70% Ethanol 16: Temperature (G) 3.4: Filtering B: Filtration Process 4.3: Filtrate (1) 4.4: Filtrate (2) C: Drying Process 5.3: Temperature (°C) 6.4: Black Poplar Propolis Aqueous Extract E: Lyophilization Process 9: Temperature (E) 10: Maltodextrin 17: Black Poplar Propolis Extract 18: Mixture of green tea extract (6.1), blueberry extract (11), pomegranate peel extract (14) and black poplar propolis extract (17) Disclosure of the Invention This invention relates to a mixture of extracts of green tea (6.1), blueberry (11), pomegranate peel (14) and black poplar propolis (17) that targets host proteins, prevents viral infection, and reduces the activity of the SARS-CoV-2 virus transmitted through the upper respiratory tract. The invention has been developed in the form of a throat lozenge / chewable tablet to provide a protective effect around the mouth and throat, which is the first entry point of the SARS-CoV-2 virus. The efficacy of the mixture against the SARS-CoV-2 virus was tested in vitro on VERO E6 cells. At the end of the 160-hour experiment, the mixture neutralized the virus at a concentration of 110 pg / mL at a rate of 112% (61% cell viability) and at a concentration of 70 pg / mL at a rate of 35% (82% cell viability). The invention describes the development of a throat lozenge / chewable tablet, as shown in Example-2. The following excipients are used in the present invention: sorbitol (E420), ascorbic acid (E300), citric acid (E330), tartaric acid (E334), magnesium salts of fatty acids (E470b), microcrystalline cellulose (E4601), maltodextrin, gum arabic (E414), lecithin, bovine gelatin (E441), polyethylene glycol (E1521), glycerin, carboxymethyl cellulose (E466), hydroxypropyl cellulose (E463), lactose, sucrose, sucralose (E955), sucrose, xylose, glycerol (E422), sodium chloride, natural flavorings (such as mint, lemon flavor), and natural coloring agents (such as beta-carotene, lycopene). In the present invention, green tea (6.1), blueberry (11), pomegranate peel (15) and Black Poplar propolis (17), whose antiviral activities have been proven by in vitro tests, are formulated in specific ratios after liquid extraction using water and alcohol solution and drying. The invention, when used against the SARS-CoV-2 virus, has the ability to neutralize 112% of the virus at a concentration of 110 µg / mL, and cell viability was found to be 61% at this concentration. Obtaining the Extracts: Green Tea Extract (6.1): Green tea is ground into a fine powder and mixed with 100% ethanol (1) in a water bath at 60-95 °C (2) (preferably 85 °C) for 0.5-2 hours (preferably 1 hour) to obtain a solid:liquid ratio of 1:21:20 (w:v) (preferably 1:10, w:v). After the mixture reaches room temperature, it is washed with 100% ethanol (1) while passing it through a filter (3.1). Then, the ethanol (1) in the filtrate (4.1) is evaporated under vacuum at 40 °C (5.1) until it is dry. Green Tea Extract (6.1) is obtained. Techniques such as supercritical liquid extraction, pressurized liquid extraction, accelerated solvent extraction, and aqueous extraction, but not limited to these, can be used as alternatives to solid / liquid extraction in the production of Green Tea Extract (6.1). Blueberry Extract (11): After grinding frozen blueberry fruit, it is mixed with 30-70% ethanol (7) (preferably 50%) solvent at 20-50 °C (8) (preferably room temperature) for 0.5 - 2 hours (preferably 1 hour) to obtain a solid:liquid ratio of 1:2 - 1:40 (w:v) (preferably 1:10, w:v). Then, the mixture is washed with 100% ethanol (1) while passing through a filter (3.2), and the ethanol (1) in the filtrate (4.2) is evaporated under vacuum at 40 °C (5.2). The blueberry aqueous extract (6.2) is mixed with Maltodextrin (10) and subjected to lyophilization starting from -40 °C (9) for 72 hours to remove the remaining water. After process (E), blueberry extract (11) is obtained. In obtaining (11), techniques such as supercritical liquid extraction, pressurized liquid extraction, accelerated solvent extraction, aqueous extraction, and liquid extraction can be used as alternatives to liquid extraction, but not limited to these. Pomegranate Peel Extract (14). After the dried pomegranate peels are ground into a fine powder, they are mixed with 25-75% ethanol (12) (preferably 30%, v / v) solvent at 30-70 °C (13) (preferably 50 °C) for 15 minutes - 2 hours (preferably 30 minutes) in a water bath with a solid-liquid ratio of 1:2 - 1:20 (preferably 1:10, w:v). After the mixture reaches room temperature, it is washed with 100% ethanol (1) while passing through a filter (3.3), and the ethanol (1) in the filtrate (4.3) is evaporated under vacuum at 40 °C (5.2). Pomegranate peel aqueous extract (6.3) is mixed with maltodextrin (10) and subjected to lyophilization for 72 hours starting at -40 °C (9) to remove the remaining water. After process (E), extract (14) is obtained. In obtaining (14), techniques such as supercritical liquid extraction, pressurized liquid extraction, accelerated solvent extraction, and aqueous extraction can be used as alternatives to liquid extraction, but not limited to these. Black Poplar Propolis Extract (17): After the raw black poplar propolis is ground into a fine powder, it is mixed with 50-96% ethanol (15) (preferably 70%, v / v) solvent at 20-50 °C (16) (preferably 24 °C) for 2-4 hours (preferably 3 hours) with a solid-liquid ratio of 1:2 - 1:20 (preferably 1:10, w:v). The upper part of the mixture is carefully filtered (3.4). The same process is repeated (B). Then, the filtrates (4.3 and 4.4) are combined and the ethanol is evaporated under vacuum at 40 °C (5.3) (C). Black Poplar Water Extract (6.4) is mixed with maltodextrin (10) and subjected to lyophilization starting at -40 °C (9) for 72 hours to remove the remaining water. After process (E), extract (17) is obtained. As an alternative to liquid extraction, techniques such as supercritical liquid extraction, pressurized liquid extraction, accelerated solvent extraction, and aqueous extraction can be used to obtain (17). After separately obtaining 6.1, 11, 14 and 17, the mixture that is the subject of the invention is obtained by mixing green tea extract (6.1) in amounts ranging from 20-100 mg, blueberry extract (11) in amounts ranging from 35-100 mg, pomegranate peel extract (14) in amounts ranging from 20-50 mg, and black poplar propolis extract (17) in amounts ranging from 35-75 mg in a chewable tablet / lozenge. The resulting mixture is converted into its final forms of lozenges and chewable tablets by adding auxiliary substances. While being converted to its final form, the mixture can also be converted into soft gel, tablet, throat spray, etc. forms, but not limited to these. V Protective lozenge / chewable tablet against SARS-CoV-2 virus contains 10-55% by weight green tea extract (6.1), 10-50% by weight blueberry extract (11), 10-55% by weight pomegranate peel extract (15), 10-50% by weight black poplar propolis extract (17) and excipients. The weight ratios of blueberry extract, pomegranate peel extract, and black poplar propolis extract in V Pastil / chewable tablet are 3(± 1.5):4(±2):3(±1.5):4(±2), respectively. Table 1. Extracts used in the mixture effective against SARS-CoV-2 and sample formulation. Code Plant Origin Extraction Conditions Active ingredient amount (minimum %) Per unit % 6.1 Green Tea Camellia sinensis Solid:liquid ratio 1:10 (w:v), 85°C, 1 hour, 100% ethanol EGCG 10% 10-55 11 Blueberry Vaccinium myrtillus Solid:liquid ratio 1:10 (w:v), room temperature, 1 hour, 50% ethanol Total Anthocyanin 0.75% 10-50 14 Pomegranate Peel Punica granatum Solid:liquid ratio 1:10 (w:v), 50°C, 30 minutes, 30% ethanol EA* 0.2% 10-55 17 Black Poplar propolis Populus nigra propolis Solid:liquid ratio 1:10 (w:v), room temperature, 3 hours, 70% ethanol CAFE 0.65% 10-50 ★Ellagic acid (EA) is the result of the ellagic acid equivalent of the components punlcalln, valoneic acid dilactone, galagic acid dilactone and ellagic acid. Example 1: Sample formulation of the invention: Effective chewable tablet formulation against SARS-CoV-2. Green tea extract (at least 10% EGCG) 35 mg Pomegranate peel extract (at least 0.2% EA*) 46 mg Blueberry extract (0.75% anthocyanin) 35 mg Black poplar propolis extract (at least 0.65% CAFE) 46 mg Sorbitol (E420) 350 mg Microcrystalline cellulose (E4601) 29 mg Maltodextrin 22 mg Flavoring (Menthol) 0.52 mg Magnesium salts of fatty acids (E470b) 9 mg Polyethylene glycol (E1521) 4 mg Sucralose 0.29 mg ★Ellagic acid (EA) is the result of punicalin, valoneic acid dilactone, galagic acid dilactone, and ellagic acid components as ellagic acid equivalents. Example 2: In vitro experiments Antiviral efficacy testing is performed impedance-based using the xCELLigence MP real-time cell analyzer. In this study, VERO E6 cells (ATCC CRL 1586) with ACE2 receptors to which SARS-CoV-2 virus binds were used, DMEM as the culture medium and 10% fetal bovine serum (FBS, heat inactivated, Hyclone) + 1% antibiotic-antifungal solution as additives [2-4]. VERO E6 cells are inoculated onto a disposable, sterile 96-well E-plate cell tray of the xCELLigence RTCA MP device, with 25,000 cells per well. The device is kept inside the incubator throughout the experiment, and the inoculated cells are placed in the device and grown for 24 hours. Samples prepared according to the sample preparation protocol are incubated with 3.5 x 10⁵ PFU / mL SARS-CoV-2 for 1 hour at 37°C with 5% CO₂. Following incubation, the samples are added to the wells after removing the culture medium from the cells in the cell growth tray, and incubated for 160 hours at 37°C with 5% CO₂. Data collected at 15-minute intervals are analyzed using RTCA Software Pro. Figure 2 shows the graph of the neutralization efficacy of a throat lozenge / chewable tablet sample against SARS-CoV-2 virus on VERO E6 cells obtained from the xCELLigence RTCA MP device. The sample and virus were added to the cells at 24 hours, and the experimental results were normalized to the previous reading range. The experiment continued for 160 hours. In the graph, (-+) media control (cells / virus only), (--) throat lozenge / chewable tablet containing 110 pg / mL virus, ) Throat lozenges / chewable tablets containing 55 pg / mL of virus and (x) virus control (3.5x105 It has been shown that at a concentration of 110 pg / mL, the throat lozenge / chewable tablet neutralizes the virus by 112% (with 61% cell viability). How the invention can be applied to industry. This invention can be easily produced and commercialized by any company operating in the pharmaceutical and food industries, and offered for sale as a dietary supplement in pharmacies or supermarkets. References: [1] World Health Organization, www.who.int. (Accessed 12 / 07 / 2021). [2] Witkowski P., Schuenadel L., Wiethaus, J. 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[21] Yeşilada E. Apiterapi: Arılardan Gelen Sağlık. Hayy Yayınları, Istanbul (2015).

Claims

17 REQUESTS 1. It is a protective lozenge / chewable tablet against the SARS-CoV-2 virus; its characteristics are as follows: by weight... 10-55% green tea extract (6.1), 10-50% blueberry extract (11) by weight 10-55% pomegranate peel extract (14), 10-50% by weight Black poplar propolis extract (17) and contains auxiliary substances. 5 2. A lozenge / chewable tablet, as in Claim 1 or Claim 2, containing green tea. extract, blueberry extract, pomegranate peel extract and Black Poplar propolis The weight ratios of the extracts are 3(±1.5):4(±2):3(±1.5):4(±2) respectively.

3. A lozenge / chewable tablet as in Claim 1, with a dosage of green tea per unit. 20-100 mg for extract, 35-100 mg for blueberry extract, 10 mg for pomegranate peel extract The recommended dosage is 20-50 mg for the propolis extract and 35-75 mg for the black poplar propolis extract.

4. A lozenge / chewable tablet as described in Claim 1, characterized by the following excipient: sorbitol (E420), ascorbic acid (E300), citric acid (E330), tartaric acid (E334), fat magnesium salts of acids (E470b), microcrystalline cellulose (E460i), maltodextrin, Gum arabic (E414), lecithin, bovine gelatin (E441), polyethylene glycol (E1521), glycerin, 15 carboxymethyl cellulose (E466), hydroxypropyl cellulose (E463), lactose, sucrose, sucralose (E955), sucrose, xylose, glycerol (E422), sodium chloride, natural flavorings (mint) and at least natural coloring agents (such as beta-carotene, lycopene) such as lemon flavor. It includes one or more of them.

5. The manufacturing method and characteristic of the lozenge / chewable tablet in Claim 1 is; green tea 20 extract (6.1), blueberry extract (11), pomegranate peel extract (14) and Black Mixing poplar propolis extract (17) in the proportions specified in request 1 (18), obtained Adding (18) auxiliary substances to the resulting mixture and making the mixture a lozenge / chewable tablet It involves the steps involved in converting it to its current form.

6. The production method in claim 5, its characteristic is; 25 -Green tea extraction (A) -Filtered (B) -Drying (C) 18 -Blueberry extraction (D) - Lyophilization (E) -Pomegranate Peel Extraction (F) -Black Poplar Propolis Extraction (G) It includes the steps of the process. 5 7. The production method described in Claim 6 is characterized by: Green tea extraction (A), Filtering (B) and The drying (C) process steps are used to obtain green tea extract (6.1).

8. The production method in claim 6 is characterized by: Blueberry extraction (D), Filtering. (B), Drying (C) and Lyophilization (E) process steps of blueberry extract (11) It is obtained. 10 9. The production method in Claim 6 is characterized by: Pomegranate Peel Extraction (F), Filtering (B), Obtaining pomegranate peel extract (14) by drying (C) and lyophilization (E) process steps It is done.

10. The production method in Claim 6 is characterized by: Black Poplar Propolis Extraction (G) Black Poplar Propolis 15 with Filtration (B), Drying (C) and Lyophilization (E) process steps is the production of its extract (17).

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