Plant component composition and crude drug composition for suppressing coronavirus infection

Abstract

To provide a plant ingredient composition and a herb composition for suppressing coronavirus infection.SOLUTION: A plant ingredient composition and a herb composition according to the present invention respectively include Camellia, lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea and Angelica keiskei. The plant ingredient composition and herb composition can suppress coronavirus infection in host cells.SELECTED DRAWING: Figure 1(A)

Description

Technical Field

[0001] The present invention relates to a plant component composition, and particularly to a plant component composition for suppressing coronavirus infection and a Kampo medicine composition containing such plant components.

Background Art

[0002] Coronaviruses are ribonucleic acid (RNA) viruses with an envelope and infect mammals and birds. Currently, known coronaviruses that infect human cells include human coronavirus 229E (hereinafter, HCoV-229E), severe acute respiratory syndrome coronavirus (hereinafter, SARS-CoV), Middle East respiratory syndrome coronavirus (hereinafter, MERS-CoV), severe acute respiratory syndrome coronavirus 2 (hereinafter, SARS-CoV-2), and the like. These coronaviruses can cause colds or serious illnesses.

[0003] Coronavirus disease 2019 (hereinafter, COVID-19) is an acute pneumonia caused by SARS-CoV-2. The main routes of infection of SARS-CoV-2 are droplet infection and contact infection, and SARS-CoV-2 is currently causing a global pandemic. Symptoms commonly seen in COVID-19 patients include fever, dry cough, fatigue, shortness of breath, muscle pain, headache, sore throat, diarrhea, and the like. Current treatment methods for COVID-19 mainly rely on supportive therapy. If there are other strategies for effectively preventing COVID-19 infection, especially those utilizing natural resources such as plants and Kampo medicines, it would be very useful.

Summary of the Invention

Problems to be Solved by the Invention

[0004] In view of the drawbacks of the foregoing prior art, the main object of the present invention is to provide a plant component composition and a Kampo medicine composition having specific plant components for suppressing coronavirus infection.

Means for Solving the Problems

[0005] To achieve the above object, the plant component composition of the present invention provides a plant component composition including camellia, lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei.

[0006] The plant component composition according to an embodiment of the present invention includes 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei.

[0007] The plant component composition according to an embodiment of the present invention further includes lotus seed of Nelumbo nucifera and Trigonella foenum - graecum.

[0008] The plant component composition according to an embodiment of the present invention includes 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, 2 parts by weight of Angelica keiskei, 1 part by weight of lotus seed of Nelumbo nucifera, and 1 part by weight of Trigonella foenum - graecum.

[0009] To achieve the above object, the crude drug composition for suppressing coronavirus infection of the present invention includes camellia, lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei.

[0010] The crude drug composition according to an embodiment of the present invention includes 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei.

[0011] The crude drug composition according to an embodiment of the present invention further includes lotus seed of Nelumbo nucifera and Trigonella foenum - graecum.

[0012] The crude drug composition according to an embodiment of the present invention comprises 3 parts by weight of Camellia japonica, 3 parts by weight of the seedpod of Nymphaea tetragona, 2 parts by weight of Pueraria lobata, 3 parts by weight of Saururus chinensis, 2 parts by weight of Angelica keiskei, 1 part by weight of the fruit of Nymphaea tetragona, and 1 part by weight of Broussonetia kazinoki.

[0013] The crude drug composition according to an embodiment of the present invention is used for preventing the invasion of coronavirus, suppressing syncytium formation, and limiting the syncytium formation area.

Effects of the Invention

[0014] In response to the above, the plant component composition or the crude drug composition containing Camellia japonica, the seedpod of Nymphaea tetragona, Pueraria lobata, Saururus chinensis, and Angelica keiskei is used to suppress the infection of coronavirus to host cells.

Brief Description of the Drawings

[0015]

Figure 1(A)

Figure 1(B)

Figure 2

Figure 3(A)

Figure 3(B)

Figure 3(C)

Modes for Carrying Out the Invention

[0016] As used in this specification, the terms "one embodiment" and "in one embodiment" mean that the described embodiment may include a particular appearance, feature, structure, or property, but do not limit that all embodiments must include that particular appearance, feature, structure, or property. Further, these terms may refer to the same embodiment mentioned in other parts of this specification, but not necessarily so. Also, when a particular module, appearance, feature, structure, or property is described and incorporated into one embodiment, whether or not there is a clear description in this specification, those skilled in the art can combine that module, appearance, feature, structure, or property with other embodiments. That is, any module, element, or feature can be combined with other elements and features in different embodiments, unless they have clearly or essentially incompatible properties or are clearly excluded.

[0017] In this embodiment, two herbal medicine compositions (designated as RXC19-A and RXC19-B, respectively) that can effectively prevent the infection of coronavirus and its entry into host cells are provided. First, the herbal medicine composition RXC19-A contains the plant components of camellia, lotus seedpod, Chinese quince, Japanese raisin tree root, and ashitaba. The herbal medicine composition RXC19-B contains the plant components of camellia, lotus seedpod, Chinese quince, Japanese raisin tree root, ashitaba, lotus fruit, and hovenia dulcis. Further, the plant components of camellia, lotus seedpod, Chinese quince, Japanese raisin tree root, ashitaba, lotus fruit, and hovenia dulcis can also be used as herbal medicines. Therefore, the herbal medicine compositions RXC19-A and RXC19-B are also the plant component compositions RXC19-A and RXC19-B described in the present invention, and are hereinafter referred to as composition RXC19-A and composition RXC19-B. Hereinafter, first, the preparation of the composition RXC19-A and the composition RXC19-B of the present invention will be described, and the infection inhibitory effect and the virus entry inhibitory effect on host cells of the composition RXC19-A and the composition RXC19-B of this embodiment will be exemplified.

[0018] Preparation of the crude drug composition of this embodiment: First, prepare a plurality of types of plant components including camellia, lotus seedpods, Chinese torreya, oyster plant, ashitaba, lotus fruits, and elm leaves.

[0019] Next, dry the above plant components to obtain dried camellia, lotus seedpods, Chinese torreya, oyster plant, ashitaba, lotus fruits, and elm leaves. In this embodiment, a low-temperature drying process may be used. Finally, for the crude drug composition RXC19-A of this embodiment, the above plant components are mixed at a ratio of 3 parts by weight of camellia, 3 parts by weight of lotus seedpods, 2 parts by weight of Chinese torreya, 3 parts by weight of oyster plant, and 2 parts by weight of ashitaba. For the crude drug composition RXC19-B of this embodiment, the above plant components are mixed at a ratio of 3 parts by weight of camellia, 3 parts by weight of lotus seedpods, 2 parts by weight of Chinese torreya, 3 parts by weight of oyster plant, 2 parts by weight of ashitaba, 1 part by weight of lotus fruit, and 1 part by weight of elm leaf.

[0020] Preparation of samples of the above plant components: The above dried plant components (i.e., dried camellia, lotus seedpods, Chinese torreya, oyster plant, ashitaba, lotus fruits, and elm leaves) are each extracted 3 times with ethanol at room temperature for 3 days each. Their ethanol solutions are concentrated under reduced pressure to obtain ethanol extracts, i.e., crude extracts. The dried crude extracts are dissolved in dimethyl sulfoxide (DMSO) at a concentration of 100 mg / mL and sonicated at 25°C for 30 minutes. Thereafter, undissolved residues are removed by filtration using a centrifuge, and the supernatant becomes the samples of these plant components. In the following description and figures of the present invention, the sample of camellia is called LCV01, the sample of lotus fruit is called LCV02, the sample of lotus seedpod is called LCV03, the sample of Chinese torreya is called LCV04, the sample of oyster plant is called LCV05, the sample of ashitaba is called LCV06, and the sample of elm leaf is called LCV07.

[0021] Furthermore, the composition RXC19-A is a mixture of five plant components, namely LCV01 (Camellia japonica), LCV03 (seed pod of Acorus calamus), LCV04 (Euonymus alatus), LCV05 (Dioscorea bulbifera), and LCV06 (Aster tataricus), and the weight ratio is 3:3:2:3:2. The composition RXC19-B is a mixture of seven plant components, namely LCV01 (Camellia japonica), LCV03 (seed pod of Acorus calamus), LCV04 (Euonymus alatus), LCV05 (Dioscorea bulbifera), LCV06 (Aster tataricus), LCV02 (fruit of Acorus calamus), and LCV07 (Acer mono), and the weight ratio is 3:3:2:3:2:1:1.

Example

[0022] Example 1 of pseudovirus neutralization assay: In this example, a pseudovirus neutralization assay was used to observe whether the plant components LCV01-LCV07, the composition RXC19-A, and the composition RXC19-B can effectively inhibit coronavirus infection of host cells by blocking virus entry. Specifically, the life cycle of SARS-CoV-2 includes entry into host cells, replication of genomic ribonucleic acid (RNA), virus assembly, budding by exocytosis, and release from host cells. The entry of SARS-CoV-2 into host cells utilizes the binding of the spike protein on the virus envelope to the angiotensin-converting enzyme 2 (hereinafter referred to as ACE2) receptor of the host cell.

[0023] Therefore, in this example, a wild-type SARS-CoV-2 pseudotyped lentivirus expressing green fluorescent protein (hereinafter referred to as WT-GFP-SARS-CoV-2 pseudotyped lentivirus) and human fetal kidney cells 293T expressing ACE2 (hereinafter referred to as 293T cells) were used. First, 293T cells expressing ACE2 were cultured in low-serum medium (hereinafter referred to as Opti-MEM) supplemented with the plant components LCV01 to LCV07, the composition RXC19-A, and the composition RXC19-B at specific concentrations. Then, they were incubated with the WT-GFP-SARS-CoV-2 pseudotyped lentivirus to observe whether the plant components and the composition could effectively inhibit the viral entry and infection of 293T cells by the GFP-SARS-CoV-2 pseudotyped lentivirus.

[0024] Specifically, 293T cells (1×10 4 cells) stably expressing the human ACE2 gene were seeded into each well of a black 96-well plate containing 50 μL of Opti-MEM and cultured overnight at 37°C and 5% CO2. The next day, the overnight culture medium in the 96-well plate was replaced with fresh Opti-MEM containing the plant components LCV01 to LCV07, the composition RXC19-A, and the composition RXC19-B at a final concentration of 10 μg / mL each. The 293T cells were incubated with each of the plant components LCV01 to LCV07, the composition RXC19-A, and the composition RXC19-B at 37°C and 5% CO2 for 1 hour each. Note that chloroquine (0.5 μg / mL) was used as a positive control for virus entry inhibition. The solvent control was carried out by adding only the solvent vehicle without plant components, compositions, or drugs.

[0025] Next, for transduction, the WT-GFP-SARS-CoV-2 pseudotyped lentivirus was introduced into the above 96-well plate containing 293T cells at a final multiplicity of infection (MOI) of 0.1. Subsequently, 16 hours after infection, the medium was replaced with fresh DMEM (supplemented with 10% FBS, 100 U / mL penicillin, and streptomycin), and the 293T cells were further cultured for 56 hours. After culturing the infected 293T cells at 37 °C and 5% CO2 for 72 hours, the GFP fluorescence of the infected 293T cells was quantified using an ImageXpress Micro Confocal high-content imaging system. The experimental results are shown in FIGS. 1(A) and 1(B). FIGS. 1(A) and 1(B) show the results of the inhibitory effects on the entry and infection of wild-type SARS-CoV-2 pseudotyped retrovirus after treatment with the composition RXC19-A and the composition RXC19-B and their plant components according to an embodiment of the present invention. Specifically, FIG. 1(A) shows a fluorescence image, and FIG. 1(B) is a bar graph showing the quantification of fluorescence and converted into the inhibition rate of virus entry and infection.

[0026] In this example, after staining the infected cells with the fluorescent dye DAPI at 37 °C for 20 minutes, GFP-positive cells (i.e., 293T cells infected with the WT-GFP-SARS-CoV-2 pseudotyped lentivirus) and total cell nuclei were detected using an ImageXpress Micro Confocal high-content imaging system. For each well, the total cells (indicated by nuclear staining) and GFP-positive cells were quantified, and the transduction rate was determined by dividing the number of GFP-positive cells by the total number of cells. Then, the relative transduction rate was obtained by normalizing the transduction rate of the plant component, composition, or chloroquine-treated group to the transduction rate of the solvent control (i.e., only the solvent vehicle was added). As shown in FIG. 1(B), the solvent control was regarded as 0% inhibition, and the inhibition rate was obtained.

[0027] The pseudotyped virus used in this example, namely the WT-GFP-SARS-CoV-2 pseudotyped lentivirus, has the GFP gene, so GFP expression can be observed after entry into host cells. That is, infected cells can be detected by green fluorescence, similar to the fluorescence image of the untreated solvent control (i.e., solvent vehicle only) as shown in Fig. 1(A). Furthermore, Fig. 1(B) shows that the positive control treated with chloroquine (0.5 μg / mL) exhibited an inhibitory effect of approximately 70%. The virus entry inhibition rates of the plant components LCV01-LCV07 at 10 μg / mL, and the compositions RXC19-A and RXC19-B are approximately 83%, 81%, 76%, 86%, 81%, 72%, 85%, 88% and 82% respectively, as shown in Fig. 1(B). Therefore, as shown in Figs. 1(A) and 1(B), the various plant components (i.e., camellia, lotus seedpod, Chinese quince, Japanese raisin tree, ashitaba, lotus fruit, and coltsfoot) and their compositions (i.e., RXC19-A and RXC19-B) used in this embodiment can effectively prevent host cell infection by blocking virus entry.

Example

[0028] Example 2 of pseudotyped virus neutralization assay: In this example, the GFP-SARS-CoV-2 pseudotyped lentivirus was incubated with the compositions RXC19-A and RXC19-B using Opti-MEM for serial dilution of the plant components LCV01-LCV07 and pretreatment of the SARS-CoV-2 pseudotyped lentivirus (Omicron type). Subsequently, the mixture of the pseudotyped virus and the plant components was added to 293T cells for transduction, and it was observed whether the plant components and the compositions could effectively inhibit virus entry and infection, as in Example 1. The 50% inhibitory concentration (IC 50 ) of the plant components LCV01-LCV07 and the compositions RXC19-A and RXC19-B against GFP-SARS-CoV-2 (Omicron type) was calculated.

[0029] 293T cells (1×10 4 ) stably expressing the human ACE2 gene in 50 μL of Opti-MEM were seeded into each well of a black 96-well plate and cultured overnight at 37 °C under 5% CO2. The next day, plant components LCV01-LCV07 and compositions RXC19-A, RXC19-B were each serially diluted two-fold from 80 μg / mL in Opti-MEM. After six two-fold dilutions, a final concentration of 1.25 μg / mL was obtained. The plant components LCV01-LCV07 and compositions RXC19-A, RXC19-B serially diluted with Opti-MEM medium were incubated with GFP-SARS-CoV-2 pseudotype (Omicron type) lentivirus at 37 °C for 1 hour. Next, the above mixtures (i.e., the mixtures of GFP-SARS-CoV-2 pseudotype lentivirus (Omicron type) with each of the plant components LCV01-LCV07, and the mixtures of GFP-SARS-CoV-2 pseudotype lentivirus (Omicron type) with each of the compositions RXC19-A and RXC19-B) were introduced at a final multiplicity of infection (MOI) of 0.1 for transduction into the plate seeded with 293T cells expressing the ACE2 gene.

[0030] Similarly, at 16 hours post-infection, the medium was replaced with fresh DMEM (supplemented with 10% FBS, 100 U / mL penicillin and streptomycin), and the 293T cells were further cultured for 56 hours. After culturing the 293T cells at 37 °C under 5% CO2 for a total of 72 hours post-infection, the GFP fluorescence of the 293T cells was quantified using an ImageXpress Micro Confocal high-content imaging system, and the 50% inhibitory concentration (IC 50 ) of each of the plant components LCV01-LCV07 and each of the compositions RXC19-A and RXC19-B was calculated. Analysis of the GFP fluorescence intensity (i.e., calculation of the relative transduction rate and inhibition rate) was performed in the same manner as in Example 1 of the pseudotype virus neutralization assay, and the details are omitted here.

[0031] After calculating the inhibition rates of plant components LCV01 to LCV07 and compositions RXC19-A and RXC19-B against GFP-SARS-CoV-2 pseudotyped lentivirus (Omicron type) at each concentration, as shown in Fig. 2, the relationship curve between the inhibition rate and the concentration of the plant component or composition was drawn using Prism 8 software (GraphPad). Fig. 2 is a graph showing the relationship curves of the infection and virus entry inhibition effects of SARS-CoV-2 pseudotyped lentivirus (Omicron type) due to the differences in the concentrations of plant components LCV01 to LCV07, composition RXC19-A, and composition RXC19-B.

[0032] In this example, after pretreatment with the plant components LCV01 to LCV07, the composition RXC19-A, and the composition RXC19-B, the GFP-SARS-CoV-2 pseudotyped lentivirus (Omicron type) was introduced into 293T cells for transduction, and whether these plant components and compositions could inhibit the infection and entry of the GFP-SARS-CoV-2 pseudotyped lentivirus (Omicron type) was observed. As shown in Fig. 2, the increase in the concentrations of plant components LCV01, LCV03, LCV05, LCV06, LCV07, the composition RXC19-A, and the composition RXC19-B decreased the GFP expression level in 293T cells (i.e., the number of 293T cells infected with the SARS-CoV-2 pseudotyped lentivirus (Omicron type) decreased). This result suggests that the pretreatment with the plant components LCV01, LCV03, LCV05, LCV06, LCV07, the composition RXC19-A, and the composition RXC19-B can inhibit the entry of the SARS-CoV-2 pseudotyped lentivirus (Omicron type) into cells.

[0033] Next, using the non-linear regression method, the concentration at which 50% of the GFP fluorescence is expressed (IC 50 ) was determined. As shown in Table 1, this concentration represents the 50% inhibitory concentration (IC 50 ) of the components that block the entry of the SARS-CoV-2 pseudotyped lentivirus (Omicron type) (i.e., plant components LCV01 to LCV07, compositions RXC19-A and RXC19-B).

[0034]

Table 1

[0035] According to Table 1, the plant components LCV01 (camellia), LCV03 (lotus seed pod), LCV05 (oyster plant), LCV06 (ashitaba), and the compositions RXC19-A and RXC19-B have an IC 50 value below 50 μg / mL, can effectively block the entry of SARS-CoV-2 lentivirus, and suppress the infection of cells. Among them, the plant component LCV01 (camellia), the compositions RXC19-A and RXC19-B show a more excellent inhibitory effect.

Example

[0036] Example 3 regarding the observation of spike-mediated syncytium formation: In this example, it was observed whether the plant components LCV01-LCV07, the composition RXC19-A, and the composition RXC19-B have the ability to suppress spike-mediated syncytium formation. Specifically, when the spike protein on the envelope surface of SARS-CoV-2 binds to the ACE2 receptor of host cells, fusion between the bound cells and adjacent host cells is induced, and as a result, syncytia are formed and the viral genome transfer to adjacent cells is promoted. In this example, two types of cells transfected with different vectors were prepared. The first type of cell expressing the N-terminal component of the spike protein and the Venus fluorescent protein is called the effector cell. The other type of cell expressing the C-terminal component of ACE2 and the Venus fluorescent protein is called the target cell. The target cells and effector cells were co-cultured so that the spike protein on the effector cells binds to ACE2 of the target cells and syncytia are formed. At the same time, the N-terminal and C-terminal components of the Venus fluorescent protein contained in the syncytia bind to form a complete Venus fluorescent protein, emitting a green fluorescence signal for detection.

[0037] In this example, 293T cells expressing the spike protein and the N-terminal component of the Venus fluorescent protein (as effector cells) and 293T cells expressing ACE2 and the C-terminal component of the Venus fluorescent protein (as target cells) were separately prepared. The target cells and the effector cells were pretreated with the plant components LCV01-LCV07 at 20 μM, the composition RXC19-A, and the composition RXC19-B at 37 °C under 5% CO2 for 48 hours. In this example, two control groups, a "solvent control" group and a "negative control" group, were prepared. The term "solvent control" refers to the pretreatment of target cells and effector cells by adding only the solvent vehicle without adding the plant components LCV01-LCV07 or the compositions RXC19-A and RXC19-B. For the "negative control" group, the spike protein was not expressed in the effector cells and the ACE2 protein was not expressed in the target cells, respectively.

[0038] Next, the pretreated target cells and effector cells were co-cultured for 5 hours. After 5 hours of incubation, syncytia (i.e., multinucleated cells) with an enlarged green fluorescence signal were detected and calculated to analyze the number and area of syncytium formation as shown in FIGS. 3(A), 3(B), and 3(C). FIG. 3(A) is a fluorescence image of the syncytium formation result induced by the spike protein of wild-type SARS-CoV-2. FIG. 3(B) is a bar graph of the analysis of the number of syncytium formations in FIG. 3(A). And FIG. 3(C) is a bar graph of the area analysis of the syncytium formation in FIG. 3(A). In FIGS. 3(B) and 3(C), the solvent control (only the solvent vehicle) was used as a reference, and a P value of less than 0.05 (p < 0.05) was indicated by "*", a P value of less than 0.01 (p < 0.01) was indicated by "**", a P value of less than 0.001 (p < 0.001) was indicated by "***", and a P value of less than 0.0001 (p < 0.0001) was indicated by "****".

[0039] Refer to FIGS. 3(A) and 3(B). In the solvent control group, multinucleated cells showing green fluorescent signals indicating spike-mediated syncytium formation were formed. On the other hand, in the negative control group in which 293T cells did not express spike protein and ACE2 protein, no green fluorescence was detected, indicating that syncytia were not formed. When cells were pretreated with plant components LCV03 (lotus seed pods), LCV07 (kohaha), and compositions RXC19-A and RXC19-B, the number of mediated syncytia formed significantly decreased, indicating that the plant components LCV03 and LCV07 and the compositions RXC19-A and RXC19-B effectively suppressed syncytium formation.

[0040] Refer to FIGS. 3(A) and 3(C). The area of syncytium formation in the solvent control group was larger than that in the experimental group. After pretreatment with plant components LCV01 to LCV07, compositions RXC19-A, and RXC19-B, the area of syncytium formation significantly decreased. From this, it was shown that plant components LCV01 to LCV07 and compositions RXC19-A and RXC19-B can effectively limit the syncytium formation area and have the ability to suppress syncytium formation.

[0041] The above experimental results are summarized in Table 2. Specifically, the "Virus invasion prevention" column in Table 2 was tabulated based on the results in Table 1. For results where the 50% inhibitory concentration (IC 50 ) could not be calculated, or for results where IC 50 exceeded 50 μg / mL, it is shown as "no effect" "-". Results where IC 50 was more than 10 μg / mL and 50 μg / mL or less are shown as "effective" "+". IC 50The result of being 10 μg / mL or less is shown as "++" with a more excellent effect. Furthermore, the "Syncytium formation inhibition" column in Table 2 was tabulated based on the analysis result of the number of syncytia formed in Fig. 3(B). The result showing a significant difference is shown as "effect present" "+", and the result showing no significant difference is shown as "no effect" "-". The "Restriction of syncytium formation area" column in Table 2 was tabulated based on the analysis result of the syncytium formation area in Fig. 3(C). The result showing a P-value of less than 0.05 or less than 0.001 is shown as "effect present" "+", and the result showing a P-value of less than 0.0001 is shown as "more excellent effect" "++".

[0042]

Table 2

[0043] From Table 2, it can be seen that the plant components LCV01-LCV07 and the compositions RXC19-A and RXC19-B can inhibit SARS-CoV-2 virus infection through three different mechanisms (i.e., the three central columns in Table 2). Specifically, the plant components LCV01 (camellia), LCV03 (lotus seedpod), LCV05 (oleaster), LCV06 (ashitaba), and the compositions RXC19-A and RXC19-B can effectively prevent virus entry by SARS-CoV-2 pseudotyped virus and inhibit infection of cells. Among them, the plant component LCV01 (camellia) and the compositions RXC19-A and RXC19-B show better inhibitory effects. The plant components LCV03 (lotus seedpod), LCV07 (koroha), and the compositions RXC19-A and RXC19-B can effectively inhibit syncytium formation. Furthermore, the plant components LCV01-LCV07 and the compositions RXC19-A and RXC19-B can greatly limit the formation area of syncytia and effectively inhibit the proliferation of the viral genome. The effects of the three mechanisms are summarized in the "Level of inhibitory effect" column in Table 2 as the sum of the "+" signs in the three left columns. According to the "Level of inhibitory effect" column in Table 2, the compositions RXC19-A and RXC19-B are superior to single plant components in terms of the inhibitory effect on SARS-CoV-2 virus infection. That is, compared with the case of using single plant components (LCV01-LCV07), the compositions RXC19-A and RXC19-B have been shown to have the highest virus infection prevention effect. The compositions RXC19-A and RXC19-B can effectively prevent virus entry, inhibit syncytium formation, and limit the syncytium formation area.

[0044] In the above embodiments, it was exemplified that the components of RXC19-A and RXC19-B and the crude drug compositions RXC19-A and RXC19-B have the efficacy of suppressing the infection of host cells by SARS-CoV-2 pseudotyped virus. In another embodiment, the plant components and the crude drug compositions are prepared using different ratios of camellia, lotus seedpod, gambir, common bauhinia bark, and ashitaba instead of using the ratios of RXC19-A or RXC19-B, and can be applied to diseases caused by the coronavirus.

[0045] As described above, a plant component composition or a crude drug composition containing camellia, lotus seedpod, gambir, common bauhinia bark, and ashitaba can be used to suppress the infection of coronavirus to host cells.

[0046] As mentioned above, the present invention has been described based on its preferred embodiments, but it should be understood that other modifications and changes are possible without departing from the gist and scope of the present invention. 〔Appendix 1〕 A plant component composition characterized by containing camellia, the lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei. 〔Appendix 2〕 The plant component composition according to Appendix 1, characterized by containing 3 parts by weight of camellia, 3 parts by weight of the lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei. 〔Appendix 3〕 Furthermore, the plant component composition according to Appendix 1, characterized by containing the lotus seed of Nelumbo nucifera and Trigonella foenum - graecum. 〔Appendix 4〕 The plant component composition according to Appendix 3, characterized by containing 3 parts by weight of camellia, 3 parts by weight of the lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, 2 parts by weight of Angelica keiskei, 1 part by weight of the lotus seed of Nelumbo nucifera, and 1 part by weight of Trigonella foenum - graecum. 〔Appendix 5〕 A crude drug composition characterized by containing camellia, the lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei. 〔Appendix 6〕 The crude drug composition according to Appendix 5, characterized by containing 3 parts by weight of camellia, 3 parts by weight of the lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei. 〔Appendix 7〕 Furthermore, the crude drug composition according to Appendix 5, characterized by containing the lotus seed and Trigonella foenum - graecum. 〔Appendix 8〕 The crude drug composition according to Appendix 7, characterized by containing 3 parts by weight of camellia, 3 parts by weight of the lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, 2 parts by weight of Angelica keiskei, 1 part by weight of the lotus seed of Nelumbo nucifera, and 1 part by weight of Trigonella foenum - graecum. 〔Appendix 9〕 A crude drug composition for suppressing coronavirus infection, characterized by containing camellia, the lotus seedpod, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei. 〔Appendix 10〕 The crude drug composition for suppressing coronavirus infection, characterized by containing 3 parts by weight of camellia, 3 parts by weight of the lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea and 2 parts by weight of Angelica keiskei, and a crude drug composition for suppressing coronavirus infection according to Supplementary Note 9, characterized by comprising the same. A crude drug composition for suppressing coronavirus infection. [Supplementary Note 11] Furthermore, a crude drug composition for suppressing coronavirus infection according to Supplementary Note 9, characterized by comprising the fruit of Trapa japonica and Colona wagneri. A crude drug composition for suppressing coronavirus infection. [Supplementary Note 12] A crude drug composition for suppressing coronavirus infection according to Supplementary Note 11, characterized by comprising 3 parts by weight of Camellia japonica, 3 parts by weight of the seed pods of Trapa japonica, 2 parts by weight of Pterocarpus santalinus, 3 parts by weight of Zanthoxylum ailanthoides, 2 parts by weight of Angelica keiskei, 1 part by weight of the fruit of Trapa japonica, and 1 part by weight of Colona wagneri. A crude drug composition for suppressing coronavirus infection. [Supplementary Note 13] The crude drug composition is used for preventing the entry of coronavirus, suppressing syncytium formation, and restricting the syncytium formation area, and a crude drug composition for suppressing coronavirus infection according to Supplementary Note 9, characterized by the same. A crude drug composition for suppressing coronavirus infection. A crude drug composition for suppressing coronavirus infection.

Claims

1. A plant ingredient composition for suppressing coronavirus infection, characterized by comprising camellia, lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei.

2. The plant ingredient composition for suppressing coronavirus infection according to Claim 1, characterized by comprising 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei.

3. The plant ingredient composition for suppressing coronavirus infection according to Claim 1, further characterized by comprising lotus seed of Nelumbo nucifera and Trigonella foenum-graecum.

4. The plant ingredient composition for suppressing coronavirus infection according to Claim 3, characterized by comprising 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, 2 parts by weight of Angelica keiskei, 1 part by weight of lotus seed of Nelumbo nucifera, and 1 part by weight of Trigonella foenum-graecum.

5. A crude drug composition for suppressing coronavirus infection, characterized by comprising camellia, lotus seedpod of Nelumbo nucifera, Eucommia ulmoides Oliver, Glechoma hederacea, and Angelica keiskei.

6. The crude drug composition for suppressing coronavirus infection according to Claim 5, characterized by comprising 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, and 2 parts by weight of Angelica keiskei.

7. The crude drug composition for suppressing coronavirus infection according to Claim 5, further characterized by comprising lotus seed of Nelumbo nucifera and Trigonella foenum-graecum.

8. The crude drug composition for suppressing coronavirus infection according to Claim 7, characterized by comprising 3 parts by weight of camellia, 3 parts by weight of lotus seedpod of Nelumbo nucifera, 2 parts by weight of Eucommia ulmoides Oliver, 3 parts by weight of Glechoma hederacea, 2 parts by weight of Angelica keiskei, 1 part by weight of lotus seed of Nelumbo nucifera, and 1 part by weight of Trigonella foenum-graecum.

9. The crude drug composition for suppressing coronavirus infection according to Claim 5, characterized in that the crude drug composition is used for preventing the invasion of coronavirus, suppressing syncytium formation, and limiting the syncytium formation area.

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