Method for producing phytochemical compositions

Abstract

To provide a phytochemical composition that is effective in preventing infectious diseases.MEANS: A phytochemical composition is attached to the oral cavity using a polymer capable of adhering to the mucosa, and releases a phytochemical in a sustained manner. The phytochemical is one or more selected from catechins, catechin polymers, persimmon tannin, chestnut tannin, mumefural, 5-aminolevulinic acid, apigenin, allyl isothiocyanate, curcumin, gingerol, shogaols, soybean isoflavone, capsaicin, saponins, allicin, alliin, menthol, eugenol, and limonene. The site in the oral cavity to which the phytochemical composition is to be attached is the palate, the cheek, or the floor of the oral cavity.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a phytochemical composition capable of suppressing infection by pathogens and the onset of infectious diseases. Manufacturing method This is regarding. [Background technology]

[0002] A year and three months have passed since the novel coronavirus disease (COVID-19) broke out in Asian countries in December 2019, and now, more than 110 million people have been infected and over 2.5 million people have died around the world. There are still no signs of an end to the virus, and there is a growing desire for the early establishment of prevention and treatment methods.

[0003] Infectious diseases, including COVID-19, are defined as diseases in which pathogens such as viruses and bacteria invade and grow in the body, causing symptoms such as fever, diarrhea, and coughing. There are three major routes of infection for pathogens that are transmitted from person to person, such as COVID-19, influenza, Ebola hemorrhagic fever, measles, and cold syndrome: contact infection, droplet infection, and airborne infection. Through these infection routes, pathogens invade cells through the mucous membranes of the nasal cavity, oral cavity, larynx, trachea, eyes, etc.

[0004] As daily preventive measures against infectious diseases that are transmitted from person to person regardless of the type of pathogen, maintaining social distance, ventilating rooms, disinfecting, washing hands, gargling, and wearing masks are recommended. Of these, the preventive effects of maintaining social distance, ventilating rooms, and disinfecting are self-evident, but the preventive effects of washing hands, gargling, and wearing masks vary depending on conditions such as the frequency of washing hands and gargling, and the type and duration of wearing the mask, and their effectiveness is limited. For example, in multiple epidemiological studies on respiratory infections such as cold syndrome and influenza, it has been reported that hand washing with soap at key points such as before meals is effective in preventing infection (Non-Patent Document 1), and gargling with tap water or green tea several times a day is effective in preventing infection (Non-Patent Documents 2 and 3). In any case, it is impossible to end the epidemic or pandemic of highly infectious infectious diseases such as influenza, COVID-19, and Ebola hemorrhagic fever with just the effects of washing hands, gargling, and wearing masks.

[0005] In human history, numerous infectious diseases such as smallpox, measles, and influenza have caused pandemics, but these could not have been contained without the establishment of prevention through vaccines and chemotherapy using antibacterial and antiviral drugs. It goes without saying that the development and dissemination of a vaccine is essential to containing the COVID-19 pandemic, but since the vaccine is administered to healthy people who are not infected with an infectious disease and to people with underlying diseases, it must be extremely safe.

[0006] In December 2020, three types of COVID-19 vaccines developed by Pfizer (co-developed with Germany's BioNTech), Moderna, and AstraZeneca were approved for emergency use in Western countries such as the United States, the United Kingdom, and Israel, and vaccinations began. These vaccines have only been developed specifically for COVID-19 for less than a year, and there is little data in both preclinical and clinical trials, so it is difficult to say that their safety is fully guaranteed. In particular, the vaccines using mRNA from Pfizer and Moderna are the first in the world to be used clinically, and there are many unknowns. Although the clinical trial data is from thousands to tens of thousands of people, what the three COVID-19 vaccines have in common is that although they are highly effective (effectiveness rates of over 90% for both Pfizer and Moderna, and about 70% for AstraZeneca), the frequency of side effects such as anaphylactic shock, fatigue, headache, chills, nausea, vomiting, and muscle pain is several to ten times higher than that of vaccines with established safety such as influenza vaccines and measles (Non-Patent Document 4).

[0007] In terms of the effectiveness of COVID-19 vaccines, if the efficacy rate is assumed to be 90%, this means that the number of people who developed COVID-19 in the vaccinated group was one-tenth of the number of people who developed COVID-19 in the non-vaccinated group. In infectious diseases, infection and onset are clearly different stages, and the results of clinical trials so far show that vaccination reduces the number of people who develop COVID-19, but it is unclear whether it reduces the number of infected people. In other words, receiving the COVID-19 vaccines from these three companies does not eliminate the need for infection prevention measures such as maintaining social distance and wearing masks.

[0008] Furthermore, even chemotherapy drugs and vaccines that are already approved and in use may become ineffective due to genetic mutations of bacteria or viruses (estimated at 25.9 / genome / year in the case of the new coronavirus), and may have to be remade. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] Special Publication No. 56-100714 [Patent Document 2] Special Publication No. 2020-125335 [Patent Document 3] Special Report No. 2020-502171

Non-licensed documents

[0010]

Non-patent document 1

Non-patent document 2

Non-patent document 3

Non-patent document 4

[0011] As mentioned above, for infectious diseases that are transmitted from person to person, the effectiveness of everyday infection prevention methods such as hand washing, gargling, and wearing a mask is limited depending on how they are implemented, and they are not effective enough to end the epidemic or pandemic of highly contagious infectious diseases such as influenza or COVID-19. In addition, even approved chemotherapy drugs and vaccines may lose their effectiveness due to genetic mutations of bacteria or viruses. Furthermore, there is still no vaccine for COVID-19 with sufficient efficacy and safety established.

[0012] The present invention has been made to solve these problems, and an object of the present invention is to provide a phytochemical composition that can protect the oral mucosa, which is the entry route of pathogens against infectious diseases transmitted from person to person, and suppress infection by pathogens and the onset of infectious diseases, and that is highly safe and not affected by genetic mutations of pathogens. Manufacturing method The purpose of this project is to provide [Means for solving the problem]

[0013] In order to solve the above-mentioned problems, the inventors conducted intensive research and discovered that by wearing a controlled release phytochemical composition containing phytochemicals (plant-derived compounds that are not required for maintaining bodily functions and may affect health) having antibacterial and / or antiviral properties in the oral cavity, continuously supplying phytochemicals to the oral cavity and covering the entire oral mucosa (including the tongue mucosa) with phytochemicals, it is possible to exert an infection suppressing effect against pathogens that exceeds the effects of gargling and wearing a mask. Furthermore, it was discovered that this infection suppressing effect does not disappear depending on the type of pathogen or genetic mutation, which led to the completion of the present invention. Effect of the Invention

[0014] According to the present invention, a phytochemical composition is provided which is highly safe, is not affected by genetic mutations of pathogens, and is effective in preventing and treating infectious diseases transmitted from person to person. Manufacturing method is provided. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 is a photograph showing a typical state of a phytochemical composition adhering to the oral cavity one hour after the start of the test, obtained from the results of Experimental Example 1. [Diagram 2] FIG. 2 is a photograph showing a typical state of the phytochemical composition adhering to the inside of the oral cavity 6 hours after the start of the test, obtained from the results of Experimental Example 2. [Diagram 3] FIG. 3 is a graph showing the release-time curves of catechins from phytochemical compositions prepared with various polymers. [Figure 4] FIG. 4 is a photograph showing the growth inhibitory effect of a phytochemical composition on Escherichia coli. [Diagram 5] FIG. 5 is a photograph showing the viable bacterial count in saliva when the phytochemical composition was worn. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] Various drug delivery systems that supply the necessary amount of medicine to the necessary area at the necessary time have been devised to suppress side effects of medicine and increase its effectiveness. Based on this concept, the present inventors believed that in order to maximize the effect of phytochemicals having antibacterial and / or antiviral activity, it is best to continuously supply phytochemicals to the oral cavity while there is a risk of infection with pathogens, and to cover and protect the entire oral mucosa with phytochemicals, and thus invented the phytochemical composition of the present invention. That is, the phytochemical composition of the present invention has a function of being worn in the oral cavity and is designed to continuously release phytochemicals having antibacterial and / or antiviral activity.

[0017] The form of the phytochemical composition of the present invention can be selected from tablets, pills, capsules, caplets, films, or any other form that can be worn in the oral cavity.

[0018] The phytochemical composition of the present invention can be applied to the oral cavity by chemical or mechanical methods. As for the chemical method, as disclosed in JP-A-56-100714 (Patent Document 1), the phytochemical composition can be prepared using a mucoadhesive polymer and known lubricants, binders, excipients, etc., and then applied to the oral cavity.

[0019] As the polymer that imparts mucoadhesiveness to the phytochemical composition of the present invention, preferred are cellulose ethers such as hydroxypropyl cellulose or pharma- ceutically acceptable salts thereof, acrylic acid polymers such as carboxyvinyl polymers or pharma-ceutically acceptable salts thereof, ethylene glycol polymers such as polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, starch, corn protein, soy protein, guar gum, carrageenan, locust bean gum, xanthan gum, and chitosan. These polymers can be used alone or in combination of two or more. As a method for mechanically attaching the phytochemical composition of the present invention to the oral cavity, a method of coating the teeth with an apatite bone cement containing phytochemicals having antibacterial and / or antiviral activity is preferred.

[0020] In infectious diseases that are transmitted from person to person, it is important from the viewpoint of preventing infection to prevent droplets from flying from the mouth, and as a method for preventing droplets from flying, the present inventors have found that by increasing the viscosity of saliva with a thickener (the mucoadhesive polymer described in the previous paragraph can also be used as a thickener), droplets are less likely to fly. Therefore, the purpose of using a mucoadhesive polymer in preparing the phytochemical composition of the present invention is not only to adhere the phytochemical composition to the oral mucosa, but also to dissolve the mucoadhesive polymer continuously in the oral cavity and impart viscosity to the saliva.

[0021] A mucosal adhesive polymer suitable for imparting anti-splash functionality to the phytochemical composition of the present invention is one that exhibits a viscosity of a 2% aqueous polymer solution at 20°C of 100,000 mPa·s or less, preferably 15,000 mPa·s or less, and more preferably 4,000 mPa·s or less.

[0022] The phytochemical composition of the present invention is preferably worn in the oral cavity for 30 minutes or more (preferably 1 hour or more, more preferably 2 hours or more, and even more preferably 3 hours or more), and can be worn during meals. By wearing the phytochemical composition of the present invention in the oral cavity for a long period of time while in contact with people, it becomes possible to surpass the effects of commercially available phytochemical or herb-containing candies, lozenges, gargling, and wearing a mask.

[0023] The phytochemical contained in the phytochemical composition of the present invention can be any food-derived compound having antibacterial and / or antiviral properties, but it is preferable to contain one or more selected from catechins, catechin polymers, persimmon tannin, chestnut tannin, mumefural, 5-aminolevulinic acid, apigenin, allyl isothiocyanate, curcumin, gingerol, shogaol, soybean isoflavones, capsaicin, saponin, allicin, alliin, menthol, eugenol, and limonene, and it is more preferable to contain one or more selected from catechins, catechin polymers, persimmon tannin, mumefural, 5-aminolevulinic acid, apigenin, allyl isothiocyanate, curcumin, gingerol, shogaol, and soybean isoflavones.

[0024] In this specification, "catechins" is a general term that includes non-epi isomers of catechin, catechin gallate, gallocatechin, and gallocatechin gallate, which are contained in large amounts in green tea, and epi isomers of epicatechin, epicatechin gallate, epigallocatechin, and epigallocatechin gallate.

[0025] In this specification, "catechin polymers" is a general term for theaflavin, theaflavin digallate, theaflavin-3-gallate, theaflavin-3'-gallate, thearubigin, oolong theanine, and theasinensin, which are contained in large amounts in black tea, oolong tea, and the like.

[0026] The phytochemical composition of the present invention needs to be worn in the oral cavity for a long time, including during meals, during which the risk of infection by pathogens is high, and therefore the taste of the phytochemicals contained therein becomes an issue. For example, catechins, persimmon tannins, and chestnut tannins have a strong astringent taste, and it is impossible for them to remain in the oral cavity for a long time. In the prior art, when astringent components such as catechins and tannins are incorporated into throat lozenges, troches, etc., the taste is masked with sweeteners, etc., but it is also not preferable to use sweeteners in the phytochemical composition of the present invention, which is to be worn in the oral cavity for a long time.

[0027] As a result of intensive research, the present inventors have found that it is possible to reduce the astringency of both catechins or catechin polymers and persimmon tannin by mixing and grinding them. This phenomenon is presumed to be due to the formation of a complex between the molecules of catechin and tannin by mixing and grinding. In addition to mixing and grinding, the complex can be prepared by mixing catechins or catechin polymers and persimmon tannin, heating and melting them, and then cooling and solidifying them, or by dissolving catechins or catechin polymers and persimmon tannin in a solvent, and then lyophilizing or heating them to remove the solvent and co-precipitating them. The mixing ratio of catechins or catechin polymers and persimmon tannin that reduces astringency is preferably 5% to 95%, more preferably 10% to 90%, and even more preferably 25% to 75% by weight of catechins or catechin polymers.

[0028] In order to improve the astringency of catechins, catechin polymers, and persimmon tannins (hereinafter abbreviated as "catechin / tannins"), it is preferable to directly mix and grind catechins or catechin polymers with persimmon tannin. However, the astringency of catechins / tannins can also be alleviated by adding pharma- ceutically acceptable additives such as cellulose ether or acrylic acid polymers to catechins or catechin polymers and persimmon tannin. Thus, the formation of a complex between catechins or catechin polymers and persimmon tannin can be used as a method for improving the astringency of the phytochemical composition of the present invention.

[0029] The phytochemicals contained in the phytochemical composition of the present invention may be artificially chemically synthesized compounds, but may also be dried products of plants containing the phytochemicals or plant extracts (which may contain other ingredients). For example, green tea extract may be used as catechins, black tea extract as catechin polymers, persimmon tannin extract as persimmon tannin, plum extract as mumefural, dried wasabi as allyl isothiocyanate, dried turmeric as curcumin, and dried ginger as gingerol or shogaol. Furthermore, herbs such as tea tree, thyme, mint, eucalyptus, lavender, and lemon balm are widely known as plants containing phytochemicals having antibacterial and / or antiviral effects, and these extracts may be contained in the phytochemical composition of the present invention.

[0030] The amount of phytochemicals contained in the phytochemical composition of the present invention can obtain an infectious disease prevention effect by targeting in the oral cavity and sustained release of phytochemicals, even if it is less than the standard daily amount ingested as food or drink. Therefore, the product of the present invention is a phytochemical composition that has no side effects and is extremely safe. For example, the content of catechins may be 400 mg or less, which is equivalent to the amount of catechins contained in 500 mL of green tea (Oi Ocha (registered trademark; Itoen Co., Ltd.)). The content of catechin polymers may be 300 mg or less, which is the amount of theaflavins ingested when drinking 500 mL of black tea with a concentration of 3% per day. Similarly, based on the amount ingested daily from food, the content of persimmon tannin is preferably 500 mg or less as persimmon tannin extract, the content of mumefural is preferably 1000 mg or less as plum flesh extract, and the content of curcumin is preferably 50 mg or less.

[0031] The above-mentioned phytochemicals that can be contained in the phytochemical composition of the present invention are compounds whose antibacterial and antiviral effects have been clarified. In particular, catechins, catechin polymers, persimmon tannins and mumefural have both strong antibacterial and antiviral effects, and are widely commercialized as disinfectant sprays (Uji Tea Catechin Spray (registered trademark; Fujiya Tea Co., Ltd.)), throat lozenges (Theaflavin Throat Lozenges (registered trademark; Eitaro Sohonpo Co., Ltd.)), Umeno Throat Lozenges (registered trademark; Kanro Co., Ltd.)), troches (Oral Barrier (registered trademark; Nisshin Pharmaceutical Co., Ltd.)), and bad breath etiquette (Kakishibu Etiquette (registered trademark; DHC Co., Ltd.)). Furthermore, in recent years, the infection preventive effects of catechins, persimmon tannins and 5-aminolevulinic acid against the new coronavirus have been found (Non-Patent Documents 5 and 6).

[0032] The antibacterial action of catechins and catechin polymers (hereinafter abbreviated as "catechins, etc.") is due to the destruction of bacterial cell membranes, and although there is a difference in antibacterial power between gram-positive and gram-negative bacteria (the antibacterial action of catechins is stronger against gram-positive bacteria than against gram-negative bacteria), catechins, etc. exhibit antibacterial action against many bacteria such as Vibrio cholerae, Staphylococcus aureus, Salmonella, and Escherichia coli. On the other hand, the antiviral action of catechins, etc. occurs by a mechanism in which catechins, etc. bind to glycoproteins called spikes on the surface of viruses, thereby preventing the virus from infecting host cells (Non-Patent Document 7). The binding of catechins, etc. to spikes is not based on the recognition of the shape of the spikes as with antibodies induced by vaccines, but is essentially based on the property that catechins, etc. easily bind to proteins, and therefore catechins, etc. exhibit a strong antiviral action that is not affected by the type of virus or genetic mutations. Therefore, the phytochemical composition of the present invention containing catechins and the like has an infection preventive effect against various infectious diseases, and the effect is not affected by genetic mutations of the pathogens. Furthermore, since the infection preventive effect of the phytochemical composition of the present invention is based on the prevention of infection by pathogens, the phytochemical composition of the present invention is particularly effective against infectious diseases such as COVID-19, which are highly infectious in asymptomatic infected individuals.

[0033] To sustain the release of phytochemicals from the phytochemical composition of the present invention, technologies used in the development and manufacture of drug delivery systems can be used, namely, a method of controlling the diffusion rate using polymers (Patent Document 2), a method of utilizing osmotic pressure (Patent Document 3), and a method using ion exchange materials.

[0034] Polymers suitable for sustaining the release of phytochemicals from the phytochemical composition of the present invention include natural polymers and semi-synthetic polymers that impart mucoadhesiveness to the phytochemical composition of the present invention as described above. That is, cellulose ethers such as hydroxypropyl cellulose or pharma- ceutical acceptable salts thereof, acrylic acid polymers such as carboxyvinyl polymers or pharma-ceutical acceptable salts thereof, ethylene glycol polymers such as polyethylene glycol, polyvinylpyrrolidone, polyvinyl alcohol, starch, corn protein, soy protein, guar gum, carrageenan, locust bean gum, xanthan gum, and chitosan can be used. These polymers can be used alone or in combination of two or more kinds, and the release of phytochemicals can be controlled as a matrix-type or reservoir-type phytochemical composition.

[0035] The duration of phytochemical release in the phytochemical composition of the present invention is 30 minutes or more, preferably 1 hour or more, more preferably 2 hours or more, and even more preferably 3 hours or more, assuming that the phytochemical agent of the present invention is worn in the oral cavity to protect the mucosa during a period when there is a risk of pathogen infection.

[0036] The method for preventing and treating infectious diseases using the phytochemical composition of the present invention differs from conventional mucoadhesive preparations (Aftouch (registered trademark; Teijin Pharma Limited)) intended for the local treatment of oral diseases. In other words, from the viewpoint of sustained supply of phytochemicals, the location where the phytochemical composition of the present invention is applied in the oral cavity is not particularly limited as long as it is a location that will not peel off for more than 30 minutes, but is preferably the palate, cheeks, or floor of the mouth, more preferably the palate or cheeks, and even more preferably the palate.

[0037] In order to protect the oral mucosa with the phytochemicals released from the phytochemical composition of the present invention, it is necessary to actively spread the phytochemicals in the oral cavity because the amount of moisture in the oral cavity is less than that in the digestive tract. In other words, the infectious disease prevention and treatment effects of the phytochemical composition of the present invention can be enhanced by spreading the phytochemicals released from the phytochemical composition of the present invention in the oral cavity with the tongue to cover the mucosa periodically or when there is a high risk of infection.

[0038] Even in infectious diseases such as COVID-19, which have a long incubation period (average 5 to 6 days) from infection by the pathogen to onset and are contagious even during the incubation period, using the phytochemical composition of the present invention in the manner described above not only prevents infection and onset, but also kills pathogens in the saliva of infected individuals, thereby suppressing the spread of droplet infection.

[0039] The present invention will be described in more detail below by way of experimental examples, but the present invention is not limited to these.

[0040] <Experimental Example 1: Evaluation of intraoral attachment site> A tablet-type phytochemical composition prepared using a mucoadhesive polymer was evaluated for its placement in the oral cavity.

[0041] (Preparation of Phytochemical Compositions) Persimmon tannin (persimmon tannin powder (Acrosia Co., Ltd.)) was selected as a phytochemical with antibacterial and antiviral properties, and approximately 300 mg of a mixed powder of hydroxypropyl methylcellulose (HPMC), carboxyvinyl polymer (CP), and persimmon tannin in a weight ratio of 1.5:0.5:1 was compressed into flat tablets with a diameter of 10 mm by direct powder compression and used in the wearing test.

[0042] (Test Method) Three males aged between 15 and 70 were selected as subjects. For the oral attachment test of the phytochemical composition, one tablet was placed on the fingertip and pressed against one of the following locations in the oral cavity: palate, cheek, or floor of the mouth, so that it adhered to the mucous membrane. After one hour, the attachment state of the phytochemical composition was evaluated. Each subject was asked to repeat the oral attachment test three times, changing the attachment site.

[0043] (Results and Effects) Figure 1 is a typical photograph of the phytochemical composition adhering to the inside of the oral cavity one hour after the start of the test, obtained as a result of Experimental Example 1. Figure 1(A) shows the phytochemical composition when it was attached to the roof of the mouth, Figure 1(B) shows the phytochemical composition when it was attached to the cheek, and Figure 1(C) shows the phytochemical composition when it was attached to the floor of the oral cavity.

[0044] In all subjects, the phytochemical composition was attached to the palate, cheek, and floor of the mouth for more than one hour. After one hour, the phytochemical composition attached to each part of the mouth was forcibly peeled off, and the mucosal adhesion of the phytochemical composition was strongest on the palate, followed by the cheek and floor of the mouth. From these results, it was revealed that the phytochemical composition of the present invention can be attached to any part of the mouth, but when it is necessary to wear it for a long time, the palate and cheek are suitable.

[0045] <Experimental Example 2: Evaluation of oral cavity wearability> The oral cavity wearing time of three types of tablet-type phytochemical compositions 1 to 3 prepared using various mucoadhesive polymers was evaluated.

[0046] (Preparation of Phytochemical Compositions 1 to 3) As a phytochemical having antibacterial and antiviral effects, catechins (epigallocatechin gallate: epigallocatechin = 2: 1 mixture) were selected, and about 300 mg of the mixed powder containing hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), locust bean gum and carboxyvinyl polymer (CP) in the weight ratio shown in Table 1 was compressed into flat tablets with a diameter of 10 mm by direct powder compression. As comparative examples, commercially available catechin troches (Oral Barrier (registered trademark; Nisshin Pharmaceutical Co., Ltd.)) and cetylpyridinium chloride troches (Pitas (registered trademark; Taiho Pharmaceutical Co., Ltd.)) were used.

[0047] [Table 1]: Weight ratio of polymers and phytochemicals contained in phytochemical compositions 1 to 3 used in Experimental Example 2 JPEG0007611783000001.jpg37169

[0048] (Test Method) The subjects were three men and two women aged between 15 and 70 years old. The five subjects were asked to place one tablet of each of the three types of phytochemical compositions on their fingertips, press it against the palate in the mouth, and attach it to the mucous membrane, and evaluate the condition and time of wearing it in the mouth. During the test, each subject had their usual lunch between 2 and 3 hours after the attachment of the phytochemical composition, and was free to drink any type of beverage. As a comparative example, the five subjects were given catechin lozenges or cetylpyridinium chloride lozenges, and the dissolution time in the mouth was measured.

[0049] (Results and Effects) Figure 2 shows typical photographs of phytochemical compositions adhering to the oral cavity 6 hours after the start of the test, obtained as a result of Experimental Example 2. Figure 2(A) shows the case where phytochemical composition 1 was worn, Figure 2(B) shows the case where phytochemical composition 2 was worn, and Figure 2(C) shows the case where phytochemical composition 3 was worn.

[0050] In all subjects, the comparative catechin troche and cetylpyridinium chloride troche were completely dissolved within 15 minutes and 23 minutes after administration, respectively, whereas the three types of phytochemical compositions 1 to 3 retained the shape of a tablet after 6 hours, and none of the subjects had the phytochemical composition peeled off from the oral mucosa. After that, phytochemical composition 3 peeled off from the oral mucosa of one subject 6.5 hours after the start of the test, and peeling was observed in the remaining four subjects after 8 hours. On the other hand, phytochemical compositions 1 and 2 remained attached to the palate while retaining the shape of a tablet for more than 8 hours (interrupted at 8 hours). From these results, it was revealed that the phytochemical composition of the present invention can be attached to the oral mucosa for a long time while maintaining the shape of the phytochemical composition without being affected by meals, and has the effect of continuously supplying phytochemicals having antibacterial and antiviral effects to the oral cavity.

[0051] <Experimental Example 3: Evaluation of phytochemical release behavior> For five types of tablet-type phytochemical compositions 4 to 8 prepared using various release control bases, the release behavior of phytochemicals from the phytochemical compositions was evaluated by in vitro dissolution tests.

[0052] (Preparation of Phytochemical Compositions) Catechins (a 2:1 mixture of epigallocatechin gallate and epigallocatechin) were selected as phytochemicals with antibacterial and antiviral properties, and about 300 mg of the mixed powder containing hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), locust bean gum, and carboxyvinyl polymer (CP) in the weight ratio shown in Table 2 was compressed into flat tablets with a diameter of 10 mm by direct powder compression. As a comparative example, about 100 mg of the catechin powder was similarly made into flat tablets with a diameter of 10 mm and subjected to the test.

[0053] [Table 2]: Weight ratio of polymers and phytochemicals contained in phytochemical compositions 4 to 8 used in Experimental Example 3 JPEG0007611783000002.jpg55168

[0054] (Test Method) The dissolution behavior of phytochemicals from Comparative Example 1 and Phytochemical Compositions 4 to 8 was evaluated by the Japanese Pharmacopoeia 16 Dissolution Test Method 2 (paddle method) using 900 ml of water as the test solution at a test temperature of 37.0±0.5° C. The amount of phytochemicals released from Phytochemical Compositions 4 to 8 was calculated by sampling 5 ml of the test solution over time and measuring the absorbance at a wavelength of 274 nm by absorbance measurement.

[0055] (Results and Effects) Figure 3 shows the catechin release-time curves from Comparative Example 1 and Phytochemical Compositions 4 to 8 obtained as a result of Experimental Example 3, and in Figure 3, the upper curve shows the release rate of catechins from Comparative Example 1, Phytochemical Composition 4, Phytochemical Composition 5, Phytochemical Composition 6, Phytochemical Composition 7 and Phytochemical Composition 8.

[0056] In the case of Comparative Example 1 consisting of catechin powder alone, disintegration occurred within 30 minutes from the start of the dissolution test, and the catechins were dissolved at almost 100%. In contrast, in the case of phytochemical compositions 4 to 8 containing various release control bases, the release of catechins was sustained for 2 hours in phytochemical composition 4 in which the release was controlled by locust bean gum, and for more than 6 hours in phytochemical compositions 5 to 8 in which the release was controlled by HPC, HPMC, and CP.

[0057] These results demonstrate that the phytochemical composition of the present invention has the effect of releasing phytochemicals for a desired sustained period of time by incorporating one or more of cellulose ether, acrylic acid polymer, and natural polymer, which are release control bases commonly used in pharmaceutical development.

[0058] <Experimental Example 4: Evaluation of the growth inhibitory effect of phytochemical compositions on bacteria> Seven types of tablet-type phytochemical compositions 9a and 9b to 15 containing various phytochemicals were evaluated for their effect of inhibiting the growth of Escherichia coli.

[0059] (Preparation of Phytochemical Compositions) Various phytochemicals with antibacterial and antiviral properties (catechins, allyl isothiocyanate (wasabi powder), curcumin (turmeric powder), gingerol (ginger powder), persimmon tannin (persimmon tannin powder), mumefural (ume extract), herb essential oil (orange sweet)) were selected, and mixed with hydroxypropyl cellulose (HPC) and carboxyvinyl polymer (CP) in the weight ratios shown in Table 3. Approximately 300 mg of the mixed powder was compressed into flat tablets with a diameter of 10 mm by direct powder compression method to obtain seven types of tablet-type phytochemical compositions 9a, 9b to 15. As comparative examples 1a, 1b, and ac, flat tablets with a diameter of 10 mm and a weight of approximately 300 mg consisting of only polymers (HPC and CP) without containing phytochemicals were used in the growth inhibition test of E. coli.

[0060] [Table 3]: Weight ratio of polymers and phytochemicals contained in the phytochemical composition used in Experimental Example 4 JPEG0007611783000003.jpg53155

[0061] (Test Method) (1) Bacterial strain used As a pathogen model, we selected Escherichia coli (E. coli DH5α) from Gram-negative bacteria that are known to be highly resistant to catechins. (2) Culture medium and E. coli concentration used E. coli was cultured in LB liquid medium at 37°C with shaking, and the optical density (OD 600 ) and measure the OD 600 The test used was 100 μL of viable bacteria with a Bacterium titer of 2.2 or 0.55 spread onto an LB agar plate (Azunol sterile petri dish, diameter 90 mm). (3) Escherichia coli growth inhibition test Five minutes after the application of E. coli, the comparative example and the phytochemical composition (tablet) were placed on the agar medium, and 100 μl of LB liquid medium was dropped onto the top surface of the phytochemical composition to promote the release of phytochemicals from the phytochemical composition. After that, the mixture was left in an incubator at 37°C and 100% relative humidity for 14 hours, and the growth state of E. coli was observed with the naked eye and by optical microscope.

[0062] (Results and Effects) Figure 4 shows the E. coli growth inhibitory effects of the comparative examples and phytochemical compositions obtained from the results of Experimental Example 4, where Figure 4(A) shows the results for phytochemical compositions 9a and 9b and comparative example 1a, Figure 4(B) shows the results for phytochemical compositions 10, phytochemical compositions 11, phytochemical compositions 12 and comparative example 1b, and Figure 4(C) shows the results for phytochemical compositions 13, phytochemical compositions 14, phytochemical compositions 15 and comparative example 1c.

[0063] In all of the petri dishes (A), (B), and (C), the tablets of the comparative example did not affect the growth of E. coli, but significant bacterial growth inhibition zones were formed in phytochemical composition 9 containing catechins, phytochemical composition 13 containing persimmon tannin, phytochemical composition 14 containing mumefural, and phytochemical composition 15 containing orange sweet. In particular, it has been reported that catechins have a weak antibacterial effect against gram-negative bacteria such as E. coli (catechins have strong antibacterial and antiviral effects against gram-positive bacteria and viruses), but it was suggested that the antibacterial effect of catechins can be enhanced by continuously releasing catechins locally according to the present invention.

[0064] These results demonstrate that the phytochemical composition of the present invention contains phytochemicals that are effective against the pathogens of the target infectious disease, thereby continuously releasing the phytochemicals locally, exhibiting antibacterial properties and exerting an infection-suppressing effect.

[0065] <Experimental Example 5: Evaluation of the oral bactericidal effect of phytochemical compositions> Tablet-type phytochemical compositions containing various phytochemicals were evaluated for their bactericidal effects against normal oral bacteria.

[0066] (Preparation of Phytochemical Compositions) As phytochemicals with antibacterial and antiviral properties, catechins (a 2:1 mixture of epigallocatechin gallate and epigallocatechin) and persimmon tannin (persimmon tannin powder) were selected, and approximately 300 mg of the mixed powder was mixed with hydroxypropyl methylcellulose (HPMC) and carboxyvinyl polymer (CP) in the weight ratio shown in Table 4 and compressed into flat tablets with a diameter of 10 mm by direct powder compression.

[0067] [Table 4]: Weight ratio of polymers and phytochemicals contained in the phytochemical composition used in Experimental Example 5 JPEG0007611783000004.jpg33129

[0068] (Test Method) For each of the three types of phytochemical compositions, one tablet was placed on the fingertip and attached to the palate of a healthy subject. After 1 hour, about 1.0 ml of saliva was collected and centrifuged at 3,000 rpm for 20 minutes. 20 μl of the supernatant was applied to 1 / 4 sections of an LB agar plate medium (Azunol sterile petri dish, diameter 90 mm) divided into four sections using a stainless steel conlarger. After that, the plate was left for 14 hours in an incubator at 37°C and 100% relative humidity, and the number of colonies formed on the medium was counted with the naked eye and under an optical microscope to determine the number of viable bacteria in the saliva. As a comparative example, saliva from a subject who had not consumed food or drink was used. Each subject was asked to repeat the oral sterilization test three times by changing the phytochemical composition.

[0069] (Results and Effects) FIG. 5 shows the viable bacterial counts in saliva when Comparative Example 2, Phytochemical Composition 16, Phytochemical Composition 17 and Phytochemical Composition 18 were worn in the oral cavity for one hour, as obtained from the results of Experimental Example 5. From the results of Comparative Example 2, more than 30 bacteria growing on LB agar plate medium were confirmed in 20 μl of saliva. For phytochemical composition 16 and phytochemical composition 17 containing persimmon tannin, no bactericidal effect was observed against normal oral bacteria when the phytochemical composition was worn. On the other hand, for phytochemical composition 18 containing catechins, no bacterial colonies were observed at all, suggesting a strong bactericidal effect in the oral cavity.

[0070] These results demonstrate that the phytochemical composition of the present invention can be an anti-infectious disease phytochemical composition with a broad antibacterial spectrum and strong antibacterial activity by selecting catechins as the phytochemical to be contained. Due to this effect, by placing the phytochemical composition of the present invention in the oral cavity of a patient who has developed an infectious disease, pathogens in the patient's saliva are inactivated, making it possible to prevent droplet infection.

[0071] <Experimental Example 6: Invention of a method for improving the taste of astringent components> The effect of mixed grinding of catechins and persimmon tannin on the astringency of the two was investigated, and a method for improving the astringency was invented.

[0072] (Sample preparation) Catechins (epigallocatechin gallate: epigallocatechin = 2:1 mixed powder) and persimmon tannin (persimmon tannin powder (Acrosia Co., Ltd.)) were placed in an agate mortar in various ratios (catechin content 100%, 95%, 90%, 75%, 50%, 25%, 10%, 5%, 0% by weight) and mixed and ground for 5 minutes. These mixed powders were stored in a silica gel desiccator until the astringency evaluation test.

[0073] (Test Method) Four males and four females aged between 15 and 60 years old were selected as subjects. 3mg of mixed powders of catechins and persimmon tannins in various mixture ratios were weighed out onto paraffin paper and the subjects were asked to evaluate the taste. The degree of astringency was rated on a score system, with 1.0 points for strong astringency, 2.0 points for astringency, 3.0 points for slight astringency, and 4.0 points for no astringency. After evaluating each sample, the subjects rinsed their mouths with mineral water and waited at least five minutes between each.

[0074] (Results and Effects) Table 5 shows the results of the astringency evaluation test for catechins and persimmon tannin powders mixed and ground at various mixing ratios obtained in Experimental Example 6. It was found that catechins alone had the strongest astringency, and by blending persimmon tannin with them, the astringency was gradually alleviated up to a blending amount of 25%. Furthermore, by increasing the blending amount of persimmon tannin from 50% or more to 95%, a significant astringency was observed, and it was improved to a level where it was only slightly astringent. As with the case of catechins, persimmon tannin alone had a strong astringency, and many subjects also complained of a sour taste. In addition to the astringency reduction, it was found that the mixed grinding of catechins and persimmon tannin powders caused surface modification of the powder particles, improving the fluidity of the mixed powder.

[0075] These results demonstrate that mixed grinding is effective in reducing the astringency of catechins and persimmon tannins. This technology makes it possible to develop the phytochemical composition of the present invention, which can be worn in the mouth for long periods of time, including during meals, during which the risk of infection by pathogens is high, and which releases catechins and persimmon tannins without causing any discomfort.

[0076] [Table 5]: Astringency evaluation of catechins and persimmon tannin powder mixed and ground at various mixing ratios JPEG0007611783000005.jpg30153 [Explanation of symbols]

[0077] 4a,4b,4c,5: Comparative example 9a, 9b to 18: Phytochemical compositions

Claims

1. A method for producing a phytochemical composition comprising mixing and grinding catechins or catechin polymers with persimmon tannin or chestnut tannin, or dissolving them in a solvent and then evaporating the solvent to obtain a mixture, and mixing the mixture with at least one of a cellulose ether and an acrylic acid polymer.

2. 2. The method for producing a phytochemical composition according to claim 1, wherein the mixing ratio of said catechins or catechin polymers to persimmon tannin or chestnut tannin is in the range of 95:5 to 5:95 by weight.

3. A method for producing a phytochemical composition according to claim 1 or claim 2, which comprises adding one or more polymers selected from xanthan gum, guar gum, carrageenan, locust bean gum, starch, ethylene glycol polymer, polyvinylpyrrolidone, and polyvinyl alcohol.

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