Antiviral traditional chinese medicine composition, traditional chinese medicine formula granule, preparation method and application thereof

Abstract

The present application relates to a kind of traditional Chinese medicine technical field, especially to a kind of antiviral traditional Chinese medicine composition, antiviral traditional Chinese medicine formula granule and its preparation method and application.The antiviral traditional Chinese medicine composition includes the following weight parts of component: 8 parts to 18 parts of Magnolia officinalis, 10 parts to 20 parts of Trichosanthes, 5 parts to 15 parts of Scutellaria, 5 parts to 15 parts of Bupleurum, 2 parts to 10 parts of Atractylodes, 2 parts to 10 parts of Ginger, 2 parts to 10 parts of North Adenophora, 3 parts to 11 parts of Red Shaoyao.The antiviral traditional Chinese medicine composition of the present application has good antiviral effect.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, and in particular to an antiviral traditional Chinese medicine composition, antiviral traditional Chinese medicine formula granules, their preparation methods and applications. Background Technology

[0002] Humans suffer from many important infectious diseases caused by viruses. Coronaviruses are a large family of viruses known to infect the upper respiratory tract and gastrointestinal tract of mammals and birds, causing disease.

[0003] It is necessary to develop traditional Chinese medicine compositions with antiviral effects against coronaviruses. Summary of the Invention

[0004] Based on this, the main objective of the present invention includes providing an antiviral traditional Chinese medicine composition, an antiviral traditional Chinese medicine formula granule, a method for preparing the same, and its application.

[0005] The first aspect of this invention provides an antiviral traditional Chinese medicine composition, the technical solution of which is as follows:

[0006] An antiviral traditional Chinese medicine composition comprising the following components in parts by weight:

[0007] Magnolia officinalis 8-18 parts, Trichosanthes kirilowii 10-20 parts, Scutellaria baicalensis 5-15 parts, Bupleurum chinense 5-15 parts, Atractylodes lancea 2-10 parts, Zingiber officinale 2-10 parts, Adenophora stricta 2-10 parts, and Paeonia lactiflora 3-11 parts.

[0008] In some embodiments, the following components are included in parts by weight:

[0009] Magnolia officinalis 10-15 parts, Trichosanthes kirilowii 13-18 parts, Scutellaria baicalensis 8-13 parts, Bupleurum chinense 8-13 parts, Atractylodes lancea 3-8 parts, Zingiber officinale 3-8 parts, Adenophora stricta 3-8 parts, and Paeonia lactiflora 4-9 parts.

[0010] In some embodiments, the following components are included in parts by weight:

[0011] Magnolia officinalis 12-13 parts, Trichosanthes kirilowii 15-16 parts, Scutellaria baicalensis 10-11 parts, Bupleurum chinense 10-11 parts, Atractylodes lancea 5-6 parts, Zingiber officinale 5-6 parts, Adenophora stricta 5-6 parts, and Paeonia lactiflora 6-7 parts.

[0012] The second aspect of this invention provides an antiviral traditional Chinese medicine formula granule, the technical solution of which is as follows:

[0013] An antiviral traditional Chinese medicine formula granule, comprising the above-mentioned antiviral traditional Chinese medicine composition.

[0014] In some embodiments, the antiviral traditional Chinese medicine formula granules also include pharmaceutically acceptable excipients.

[0015] In some embodiments, the pharmaceutically acceptable excipients include one or more of maltodextrin, dextrin, betacyclodextrin, lactose, silicon dioxide, and magnesium stearate.

[0016] The third aspect of this invention provides a method for preparing antiviral traditional Chinese medicine formula granules, the technical solution of which is as follows:

[0017] A method for preparing an antiviral traditional Chinese medicine formula granule includes the following steps:

[0018] Take the medicinal materials of each of the above components and prepare the formulation granules of each component;

[0019] The formulation granules of each component are mixed to prepare antiviral traditional Chinese medicine formulation granules.

[0020] In some embodiments, the method for preparing the formulation particles of each component includes the following steps:

[0021] Take the medicinal materials of each component, add water to extract them separately, and obtain the extracts of each component;

[0022] The extracts of each component were concentrated to obtain concentrated solutions of each component.

[0023] Pharmaceutically acceptable excipients were added to the concentrates of each component, and the mixture was spray-dried to obtain the formulation granules of each component.

[0024] In some embodiments, the mass of the water is 3 to 8 times the mass of the medicinal material to be extracted.

[0025] In some embodiments, the extraction is performed 2 to 3 times, and the extracts from each extraction are combined to obtain a single-component extract.

[0026] In some embodiments, the relative density of the concentrates of each component is 1.05 to 1.20.

[0027] The fourth aspect of the present invention provides the use of the above-mentioned antiviral traditional Chinese medicine composition or the above-mentioned antiviral traditional Chinese medicine formula granules in the preparation of a medicament for the prevention and / or treatment of diseases mediated by coronavirus HCoV-OC43.

[0028] The fifth aspect of the present invention provides the use of the above-mentioned antiviral traditional Chinese medicine composition or the above-mentioned antiviral traditional Chinese medicine formula granules in the preparation of a medicine for the prevention and / or treatment of diseases mediated by the novel coronavirus SARS-CoV-2.

[0029] Compared with traditional solutions, the present invention has the following advantages:

[0030] In the antiviral traditional Chinese medicine composition of the present invention, Trichosanthes kirilowii clears heat and eliminates phlegm, relieves chest congestion and disperses nodules; Scutellaria baicalensis clears heat and dries dampness, drains fire and detoxifies, and the two are the principal herbs; Magnolia officinalis dries dampness and eliminates phlegm, lowers qi and relieves fullness; Adenophora stricta nourishes yin and clears the lungs, eliminates phlegm and stops cough; Atractylodes lancea dries dampness and strengthens the spleen, dispels wind and cold; Paeonia lactiflora clears heat and cools blood, disperses blood stasis and relieves pain, assisting the principal herbs in clearing heat, drying dampness and eliminating phlegm, and together they are the assistant herbs; Ginger and Bupleurum chinense have the effects of relieving exterior and interior symptoms, and are the adjuvant herbs; the eight herbs are used together to achieve the effects of clearing heat and detoxifying, drying dampness and eliminating phlegm and moistening the lungs. Experimental verification shows that the antiviral traditional Chinese medicine composition of the present invention has an inhibitory effect on cytopathic effects caused by SARS-CoV-2 infection of VeroE6 cells, and can improve the weight loss caused by HCoV-OC43 coronavirus and SARS-CoV-2 infection in suckling mice. It also has a significant effect on improving the increase in brain and lung indices caused by viral infection in suckling mice, and shows a significant inhibitory effect on viral proliferation in the brain and lung tissues of suckling mice, and no toxic side effects were found. Attached Figure Description

[0031] To more clearly illustrate the technical solutions in the embodiments of the present invention and to more completely understand the present invention and its beneficial effects, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 The results of cell assays for the efficacy of comparative examples 9-11 against the novel coronavirus (SARS-CoV-2) were presented.

[0033] Figure 2 The results of cell assays for the efficacy of drugs against the novel coronavirus (SARS-CoV-2) in Examples 1-6 are shown.

[0034] Figure 3 A comparison of weight changes in OC43-infected suckling mice after treatment in different groups;

[0035] Figure 4 A comparison of brain index changes in OC43-infected suckling mice after different treatment groups;

[0036] Figure 5 A comparison of lung index changes in OC43-infected suckling mice after treatment in different groups;

[0037] Figure 6 A comparison of viral load in the brain and lung tissues of OC43-infected suckling mice after treatment in different groups. Detailed Implementation

[0038] The present invention will be further described in detail below with reference to specific embodiments. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0039] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0040] the term

[0041] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0042] In this invention, the selection range of "and / or", "or / and", and "and / or" includes any one of two or more related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected using at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that the technical solution undoubtedly includes technical solutions connected by "logical AND", and also undoubtedly includes technical solutions connected by "logical OR". For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0043] In this invention, terms such as "multiple", "various", "multiple times", and "multi-dimensional" are used, unless otherwise specified, to refer to a quantity greater than or equal to 2. For example, "one or more" means one or more types.

[0044] In this invention, the terms "combinations thereof", "any combination thereof", and "any combination thereof" include all suitable combinations of any two or more of the listed items.

[0045] In this invention, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this invention, solve the technical problem of this invention, and achieve the expected technical effect of this invention.

[0046] In this invention, terms such as "preferred," "better," "more suitable," and "ideal" are used only to describe implementation methods or embodiments with better effects, and should be understood not to limit the scope of protection of this invention.

[0047] In this invention, terms such as "further," "even more," and "particularly" are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of this invention.

[0048] In this invention, the terms "optionally," "optionally," and "optional" refer to options that are optional, meaning they are selected from either "with" or "without." If multiple "optional" options appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "optional" option is independent.

[0049] In this invention, the terms "first aspect," "second aspect," "third aspect," and "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," and "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0050] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0051] In this invention, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0052] Unless otherwise specified, the temperature parameters in this invention can be either constant temperature treatment or variations within a certain temperature range. It should be understood that the constant temperature treatment allows temperature fluctuations within the precision range controlled by the instrument. Fluctuations are permitted within ranges such as ±5℃, ±4℃, ±3℃, ±2℃, and ±1℃.

[0053] In this invention, percentage content refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures, unless otherwise specified.

[0054] In this invention, percentage concentrations, unless otherwise specified, refer to the final concentration. The final concentration refers to the proportion of the added component in the system after the addition of that component.

[0055] In this invention, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass-volume percentage.

[0056] This invention provides an antiviral traditional Chinese medicine composition. In one embodiment, the antiviral traditional Chinese medicine composition comprises the following components in parts by weight:

[0057] Magnolia officinalis 8-18 parts, Trichosanthes kirilowii 10-20 parts, Scutellaria baicalensis 5-15 parts, Bupleurum chinense 5-15 parts, Atractylodes lancea 2-10 parts, Zingiber officinale 2-10 parts, Adenophora stricta 2-10 parts, Paeonia lactiflora 3-11 parts.

[0058] In the above-mentioned antiviral traditional Chinese medicine composition, Trichosanthes kirilowii clears heat and eliminates phlegm, relieves chest congestion and disperses nodules; Scutellaria baicalensis clears heat and dries dampness, drains fire and detoxifies, and the two are the principal herbs; Magnolia officinalis dries dampness and eliminates phlegm, lowers qi and relieves fullness; Adenophora stricta nourishes yin and clears the lungs, eliminates phlegm and stops cough; Atractylodes lancea dries dampness and strengthens the spleen, dispels wind and cold; Paeonia lactiflora clears heat and cools blood, disperses blood stasis and relieves pain, assisting the principal herbs in clearing heat, drying dampness and eliminating phlegm, and together they are the assistant herbs; Ginger and Bupleurum chinense have the effects of relieving exterior and interior symptoms, and are the adjuvant herbs; the eight herbs used together have the effects of clearing heat and detoxifying, drying dampness and eliminating phlegm and moistening the lungs. Experimental verification shows that the antiviral traditional Chinese medicine composition of the present invention has an inhibitory effect on cytopathic effects caused by SARS-CoV-2 infection of VeroE6 cells, and can improve the weight loss caused by HCoV-OC43 coronavirus and SARS-CoV-2 infection in suckling mice. It also has a significant effect on improving the increase in brain and lung indices caused by viral infection in suckling mice, and shows a significant inhibitory effect on viral proliferation in the brain and lung tissues of suckling mice, and no toxic side effects were found.

[0059] Understandably, the weight parts of Magnolia officinalis in the antiviral traditional Chinese medicine composition include, but are not limited to, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 parts, and any value within any two of the above ranges.

[0060] The weight parts of Trichosanthes kirilowii in the antiviral traditional Chinese medicine composition include, but are not limited to, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 parts, as well as any value within any two of the above ranges.

[0061] The weight parts of Scutellaria baicalensis in the antiviral traditional Chinese medicine composition include, but are not limited to, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 parts, as well as any value within any two of the above ranges.

[0062] The weight parts of Bupleurum in the antiviral traditional Chinese medicine composition include, but are not limited to, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 parts, as well as any value within any two of the above ranges.

[0063] The weight parts of Atractylodes lancea in the antiviral traditional Chinese medicine composition include, but are not limited to, 2, 3, 4, 5, 6, 7, 8, 9, 10 parts, as well as any value within any two of the above ranges.

[0064] The weight parts of ginger in the antiviral traditional Chinese medicine composition include, but are not limited to, 2, 3, 4, 5, 6, 7, 8, 9, 10 parts, and any value within any two of the above ranges.

[0065] The weight parts of North American ginseng in the antiviral traditional Chinese medicine composition include, but are not limited to, 2, 3, 4, 5, 6, 7, 8, 9, 10 parts, as well as any value within any two of the above ranges.

[0066] The weight parts of Paeonia lactiflora in the antiviral traditional Chinese medicine composition include, but are not limited to, 3, 4, 5, 6, 7, 8, 9, 10, 11 parts, and any value within any two of the above ranges.

[0067] In some preferred examples, the antiviral traditional Chinese medicine composition comprises the following components in parts by weight:

[0068] Magnolia officinalis 10-15 parts, Trichosanthes kirilowii 13-18 parts, Scutellaria baicalensis 8-13 parts, Bupleurum chinense 8-13 parts, Atractylodes lancea 3-8 parts, Zingiber officinale 3-8 parts, Adenophora stricta 3-8 parts, Paeonia lactiflora 4-9 parts.

[0069] In some preferred examples, the antiviral traditional Chinese medicine composition comprises the following components in parts by weight:

[0070] Magnolia officinalis 12-13 parts, Trichosanthes kirilowii 15-16 parts, Scutellaria baicalensis 10-11 parts, Bupleurum chinense 10-11 parts, Atractylodes lancea 5-6 parts, fresh ginger 5-6 parts, Adenophora stricta 5-6 parts, Paeonia lactiflora 6-7 parts.

[0071] In some preferred examples, the antiviral traditional Chinese medicine composition comprises the following components in parts by weight:

[0072] Magnolia officinalis 12 parts, Trichosanthes kirilowii 15 parts, Scutellaria baicalensis 10 parts, Bupleurum chinense 10 parts, Atractylodes lancea 5 parts, fresh ginger 5 parts, Adenophora stricta 5 parts, Paeonia lactiflora 6 parts.

[0073] The present invention also provides an antiviral traditional Chinese medicine formula granule. In one embodiment, the raw materials of the antiviral traditional Chinese medicine formula granule include the above-mentioned antiviral traditional Chinese medicine composition.

[0074] Optionally, the antiviral traditional Chinese medicine formula granules may also include pharmaceutically acceptable excipients.

[0075] Optionally, the pharmaceutically acceptable excipients include one or more of maltodextrin, dextrin, betacyclodextrin, lactose, silicon dioxide, and magnesium stearate.

[0076] The antiviral traditional Chinese medicine formula granules in this embodiment have all the advantages of the above-mentioned antiviral traditional Chinese medicine composition, which will not be repeated here.

[0077] This invention also provides a method for preparing antiviral traditional Chinese medicine formula granules. In one embodiment, the method for preparing the antiviral traditional Chinese medicine formula granules includes the following steps:

[0078] Take the medicinal materials of each of the above components and prepare the formulation granules of each component;

[0079] The formulation granules of each component are mixed to prepare antiviral traditional Chinese medicine formulation granules.

[0080] Understandably, the preparation of the formulation granules for each component can be specifically referred to the methods published by the National Drug Standards of the National Medical Products Administration for the corresponding components.

[0081] In some examples, the method for preparing the formulation particles of each component includes the following steps:

[0082] Take the medicinal materials of each component, add water to extract them separately, and obtain the extracts of each component;

[0083] The extracts of each component were concentrated to obtain concentrated solutions of each component.

[0084] Pharmaceutically acceptable excipients were added to the concentrates of each component, and the mixture was spray-dried to obtain the formulation granules of each component.

[0085] Optionally, the mass of the water is 3 to 8 times the mass of the medicinal material to be extracted.

[0086] Optionally, the extraction is performed 2 to 3 times, and the extracts from each extraction are combined to obtain a single component extract.

[0087] Optionally, the relative density of the concentrate for each component is 1.05 to 1.20.

[0088] The present invention also provides the application of the above-mentioned antiviral traditional Chinese medicine composition or the above-mentioned antiviral traditional Chinese medicine formula granules in diseases mediated by coronavirus HCoV-OC43.

[0089] The present invention also provides the application of the above-mentioned antiviral traditional Chinese medicine composition or the above-mentioned antiviral traditional Chinese medicine formula granules in the disease mediated by the novel coronavirus SARS-CoV-2.

[0090] Optionally, in the above applications, the dosage of the antiviral traditional Chinese medicine composition or the above-mentioned antiviral traditional Chinese medicine formula granules is 100mg / kg / d to 260mg / kg / d, and more preferably 120mg / kg / d to 240mg / kg / d.

[0091] The following description is further illustrated with specific embodiments and comparative examples. Unless otherwise specified, the raw materials involved in the following specific embodiments and comparative examples are all commercially available. Unless otherwise specified, the instruments used are all commercially available. Unless otherwise specified, the processes involved are conventionally selected by those skilled in the art.

[0092] Example 1

[0093] This embodiment provides an antiviral traditional Chinese medicine composition, antiviral traditional Chinese medicine formula granules, and their preparation method, with the following steps:

[0094] Step 1: Refer to Table 1 and prepare the component medicinal materials according to the following weights:

[0095] The following is a traditional Chinese medicine composition for antiviral treatment: 12g Magnolia officinalis, 15g Trichosanthes kirilowii, 10g Scutellaria baicalensis, 10g Bupleurum chinense, 5g Atractylodes lancea, 5g Zingiber officinale, 5g Adenophora stricta, and 6g Paeonia lactiflora.

[0096] Step 2: Following the methods published in the National Drug Standards by the National Medical Products Administration, prepare Magnolia officinalis formula granules, Trichosanthes kirilowii formula granules, Scutellaria baicalensis formula granules, Bupleurum chinense formula granules, Atractylodes lancea formula granules, Zingiber officinale formula granules, Adenophora stricta formula granules, and Paeonia lactiflora formula granules.

[0097] The following traditional Chinese medicine formula granules were obtained by mixing Magnolia officinalis formula granules, Trichosanthes kirilowii formula granules, Scutellaria baicalensis formula granules, Bupleurum chinense formula granules, Atractylodes lancea formula granules, Zingiber officinale formula granules, Adenophora stricta formula granules, and Paeonia lactiflora formula granules to obtain antiviral traditional Chinese medicine formula granules.

[0098] Examples 2-6

[0099] Referring to Table 1 and the preparation method of Example 1, antiviral traditional Chinese medicine formula granules of Examples 2-6 were prepared.

[0100] Table 1

[0101] Magnolia officinalis 12 13 10 15 8 18 Trichosanthes kirilowii 15 16 18 10 20 10 Scutellaria baicalensis 10 11 8 13 5 15 Bupleurum 10 11 13 8 15 5 Atractylodes lancea 5 6 3 8 2 10 Ginger 5 6 3 8 2 10 North American ginseng 5 6 8 3 10 2 Red peony 6 7 9 4 11 3

[0102] Comparative Examples 1-19

[0103] Multiple Chinese herbal components were selected and classified into five categories: dampness-resolving, phlegm-resolving and cough-relieving, exterior-releasing and heat-clearing, strengthening the body and promoting diuresis, and bowel-clearing and blood-stasis-removing. See Table 2.

[0104] Table 2

[0105]

[0106]

[0107] Referring to Table 3, the components of the antiviral traditional Chinese medicine compositions of Comparative Examples 1 to 19 were obtained.

[0108] Table 3

[0109]

[0110]

[0111] Prepare the medicinal materials for each component as shown in Table 3 below, referring to the methods used in the publication of national drug standards by the National Medical Products Administration (NMPA). If there are no national drug standards published by the NMPA, refer to the methods used in the publication of standards formulated by the provincial drug regulatory authorities. Prepare the formulation granules for each component in Comparative Examples 1-19, and mix the formulation granules for each component accordingly to obtain the antiviral traditional Chinese medicine formulation granules for Comparative Examples 1-19:

[0112] Comparative Example 1

[0113] Patchouli 9g, Reed Rhizome 30g, Tangerine Peel 6g, Artemisia annua 10g, Schisandra Chinensis 10g, Prepared Licorice Root 6g, Stir-fried Atractylodes Macrocephala 10g.

[0114] Comparative Example 2

[0115] 10g of Lepidium apetalum, 10g of Magnolia officinalis, 10g of Bupleurum chinense, 6g of American ginseng, 10g of ginseng, 15g of Poria cocos, and 10g of stir-fried Atractylodes macrocephala.

[0116] Comparative Example 3

[0117] 5g of cardamom, 10g of bitter almond, 6g of dried tangerine peel, 10g of anemarrhena rhizome, 15g of forsythia, 6g of American ginseng, and 30g of coix seed.

[0118] Comparative Example 4

[0119] Magnolia officinalis 10g, Gypsum 30g, Scutellaria baicalensis 10g, Ophiopogon japonicus 15g, Adenophora stricta 9g, Cornus officinalis 15g, Paeonia lactiflora 10g.

[0120] Comparative Example 5

[0121] Atractylodes lancea 9g, bitter almond 10g, gypsum 30g, ephedra 9g, artemisia annua 10g, ginger 9g, poria cocos 15g.

[0122] Comparative Example 6

[0123] Fritillaria thunbergii 10g, Lonicera japonica 15g, Adenophora stricta 9g, Panax quinquefolius 6g, Glycyrrhiza uralensis (processed) 6g, Coix lacryma-jobi 30g, Atractylodes macrocephala (fried) 10g.

[0124] Comparative Example 7

[0125] 10g bitter almonds, 6g dried tangerine peel, 10g mulberry bark, 9g ephedra, 3g honey-processed ephedra, 9g North American ginseng, and 6g American ginseng.

[0126] Comparative Example 8

[0127] Patchouli 9g, Ephedra 9g, Anemarrhena 10g, Forsythia 15g, Cornus officinalis 15g, Poria cocos 15g, and stir-fried Atractylodes macrocephala 10g.

[0128] Comparative Example 9

[0129] Patchouli 9g, Magnolia officinalis 10g, Lonicera japonica 15g, Bupleurum chinense 10g, Ophiopogon japonicus 15g, Coix lacryma-jobi 30g, Rheum palmatum 5g.

[0130] Comparative Example 10

[0131] 6g dried tangerine peel, 10g trichosanthes fruit, 30g gypsum, 10g bupleurum root, 3g honey-processed ephedra, 9g north ginseng, 15g poria cocos, 10g red peony root.

[0132] Comparative Example 11

[0133] Atractylodes lancea 9g, Amomum tsao-ko 5g, Magnolia officinalis 10g, Phragmites communis 30g, Trichosanthes kirilowii 10g, Scutellaria baicalensis 10g, Zingiber officinale 9g.

[0134] Comparative Example 12

[0135] Atractylodes lancea 9g, Lepidium apetalum 10g, Phragmites communis 30g, Platycodon grandiflorus 10g, Anemarrhena asphodeloides 10g, Panax ginseng 10g, Paeonia lactiflora 10g, Rheum palmatum 5g.

[0136] Comparative Example 13

[0137] Atractylodes lancea 9g, Lepidium apetalum 10g, Phragmites communis 30g, Morus alba root bark 10g, Scutellaria baicalensis 10g, Lonicera japonica 15g, Forsythia suspensa 15g.

[0138] Comparative Example 14

[0139] Trichosanthes kirilowii 10g, Platycodon grandiflorus 10g, Gypsum 30g, Scutellaria baicalensis 10g, Lonicera japonica 15g, Forsythia suspensa 15g, Ginseng 10g.

[0140] Comparative Example 15

[0141] Patchouli 9g, Fritillaria thunbergii 10g, Morus alba root bark 10g, Platycodon grandiflorus 10g, Ephedra sinica 9g, Ophiopogon japonicus 15g, Cornus officinalis 15g.

[0142] Comparative Example 16

[0143] 10g of Lepidium seed, 10g of Platycodon root, 10g of Bupleurum root, 15g of Ophiopogon root, 10g of Schisandra fruit, 6g of prepared licorice root, and 5g of rhubarb.

[0144] Comparative Example 17

[0145] 10g bitter almond, 10g fritillaria thunbergii, 10g anemarrhena asphodeloides, 10g artemisia annua, 3g honey-processed ephedra, 15g cornus officinalis, 10g red peony root.

[0146] Comparative Example 18

[0147] 5g of Amomum tsao-ko, 10g of Morus alba root bark, 10g of Trichosanthes kirilowii fruit, 3g of honey-processed Ephedra sinica, 9g of fresh ginger, 10g of Schisandra chinensis fruit, 6g of prepared licorice root, and 5g of rhubarb.

[0148] Comparative Example 19

[0149] 5g of Amomum tsao-ko, 10g of Fritillaria thunbergii, 10g of Artemisia annua, 9g of fresh ginger, 10g of Schisandra chinensis, 10g of ginseng, and 30g of Coix lacryma-jobi.

[0150] Pharmacological studies

[0151] I. Evaluating the efficacy of drugs against SARS-CoV-2 at the cellular level

[0152] 1. Experimental Materials

[0153] 1.1 Drugs: Antiviral traditional Chinese medicine formula granules of Examples 1-6 and Comparative Examples 1-19, the concentrations of which are shown in Table 4.

[0154] 1.2 Cells: VeroE6 cells were preserved by the Virus Laboratory of the State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Health.

[0155] 1.3 Virus: SARS-CoV-2, titer: TCID 50 =10 -6 / 100μL, stored at -80℃ in the BSL-3 laboratory (National Key Laboratory of Respiratory Diseases, Highly Pathogenic Microorganisms Research Laboratory) of Guangzhou Customs Technology Center. Viral titer used was 100 TCID. 50 .

[0156] Table 4

[0157]

[0158] 2. Experimental Methods

[0159] (1) Add 100 μL of a 2×10⁻⁶ m³ solution to each well of a sterile 96-well culture plate. 5 VeroE6 cells / mL, cultured at 37°C with 5% CO2 for 24 hours;

[0160] (2) 100 TCID was added to the culture plate experimental group and the virus control group. 50 100 μL of virus solution per well, incubated at 37°C in a 5% CO2 incubator for 2 h for adsorption;

[0161] (3) After 2 hours, discard the cell culture medium in the 96-well culture plate; dilute the test drug to the concentrations shown in Table 4, with 3 replicates for each concentration, and add 100 μL / well of the above drug solution.

[0162] (4) Simultaneously set up cell control, blank control (solvent control) and virus control (negative control);

[0163] (5) Incubate the cells at 37°C in a 5% CO2 incubator for 3-4 days;

[0164] (6) Observe cytopathic effect (CPE) under an optical microscope. The degree of cytopathic effect is recorded according to the following 6-level standard: "-" No cytopathic effect; "live" less than 10%; "+" about 25%; "++" about 50%; "+++" about 75% of cells have cytopathic effect; "++++" more than 75% of cells have cytopathic effect.

[0165] The half-maximal effective concentration (IC50) was calculated using the Reed-Muench method or GraphPad Prism 5.0. 50 Criteria for judging efficacy: A concentration that inhibits the virus CPE by 50% is considered an effective concentration.

[0166] 3. Experimental Results

[0167] 3.1 Comparative samples 1–8 and 12–19 did not inhibit the cytopathic effect of SARS-CoV-2 infection on VeroE6 cells.

[0168] 3.2 Figure 1 The results of cell assays for the efficacy of comparative samples 9-11 against the novel coronavirus (SARS-CoV-2) are shown.

[0169] It was found that Comparative Example 9, at the maximum non-toxic concentration of 500 μg / mL, inhibited the cytopathic effect of SARS-CoV-2 infection on VeroE6 cells. Comparative Examples 10 and 11, within the concentration range of 250–500 μg / mL, also inhibited the cytopathic effect of SARS-CoV-2 infection on VeroE6 cells.

[0170] 3.3 Figure 2 The results of cell detection of the efficacy of the drug against the novel coronavirus (SARS-CoV-2) in Examples 1-6 are shown.

[0171] It is evident that Example 1, within a concentration range of 150–500 μg / mL, inhibited cytopathic effects of SARS-CoV-2 infection on VeroE6 cells. Examples 2 and 3, within a concentration range of 250–500 μg / mL, also inhibited cytopathic effects of SARS-CoV-2 infection on VeroE6 cells. Examples 4–6, at a maximum non-toxic concentration of 500 μg / mL, also inhibited cytopathic effects of SARS-CoV-2 infection on VeroE6 cells.

[0172] II. Efficacy evaluation of human coronavirus (HCoV-OC43, OC43) infecting C57BL / 6 suckling mice

[0173] 1. Experimental Materials

[0174] 1.1 Drugs

[0175] The antiviral traditional Chinese medicine formula granules in Examples 1-6, using Remdesivir (RDV), were donated by the Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences. All the above drugs were dissolved in PBS to the concentration to be tested.

[0176] 1.2 Reagents

[0177] EZNAviral RNA extraction R6874-02 kit (Omega Bio-Tek, USA) and OneStep PrimeScript TM RT-PCR Kit (Takara Bio, Japan), etc.

[0178] 1.3 Virus

[0179] The mouse lung-adapted strain of human coronavirus (HCoV-OC43, OC43) was preserved by the Clinical Virology Laboratory of Guangzhou Institute of Respiratory Health.

[0180] 1.4 Animals

[0181] SPF grade C57BL / 6 suckling mice (4-6g), within 10 days of birth, purchased from the Experimental Animal Center of Kunming Medical University.

[0182] 2 Experimental Methods

[0183] (1) The suckling mice were randomly divided into 21 groups: a normal control group, a virus group, a positive drug group (RDV, 25 mg / kg / d), and high, medium, and low dose groups (240 mg / kg / d, 120 mg / kg / d, and 60 mg / kg / d) as described in Examples 1-6, with 6 mice in each group. Except for the normal group, each group of mice was injected with 25 μL of virus solution containing 1000 LD50 (median lethal dose), while the normal control group was injected with an equal volume of PBS.

[0184] (2) Two hours after inoculation with the virus, the mice were administered the drug by gavage for four consecutive days, and the weight of the mice was recorded daily. Mice that died within 24 hours of infection were considered to have died abnormally. On the fourth day after infection, the mice were anesthetized and sacrificed. The brain and lung tissues were dissected, weighed, and the brain index and lung index were calculated as follows: organ weight (g) / body weight (g) × 100.

[0185] (3) RPIM 1640 medium (weight-to-volume ratio 1:10) was added to brain and lung tissues, homogenized, and centrifuged at 5000 g / min for 10 minutes. The supernatant was collected, and viral load was detected by qRT-PCR. Primer and probe sequences are shown in Table 5. The OC43n gene standard curve was R² = 1, Eff% = 101.70, and Copies / μL = 10^[(41.248-Ct) / 3.281]. The copy number / g was calculated to be 10^[ ... [(41.248-Ct) / 3.281] ×60×Added RPIM volume / 140 / organ weight (g), CT value >32 is negative.

[0186] Table 5

[0187] forward primer 5'-ATGTTAGCCCGATAATTGAGGACTAT-3' reverse primer 5'-AATGTAAAGATGGCCGCGTATT-3' probe FAM-5'-CATACTCTGACGGTCACA AT-3'3'-NFQ-MGB

[0188] 3. Statistical Analysis

[0189] Brain and lung indices and viral load were analyzed for normality and ANOVA using GraphPad Prism 8.0 software; compared with the viral group, *, p < 0.05, **, p < 0.01, ***, p < 0.001.

[0190] 4. Experimental Results

[0191] 4.1 Figure 3 A comparative graph showing the weight changes (n=6) of OC43-infected suckling mice after treatment in different groups.

[0192] It is evident that the weight gain of suckling mice infected with OC43 was smaller than that of the normal group, and weight loss occurred on the 3rd day after infection. The positive control RDV and the intervention of high-dose (240mg / kg / d) and medium-dose (120mg / kg / d) examples 1-6 can improve the weight loss of infected suckling mice to a certain extent. The low-dose (60mg / kg / d) examples 1-6 have no significant effect on improving the weight loss caused by virus infection in suckling mice.

[0193] 4.2 Figure 4 A comparative graph showing the changes in brain indices (n=6) of OC43-infected suckling mice after treatment in different groups.

[0194] It is evident that viral infection can cause a significant increase in the brain index of suckling mice (p < 0.05), and treatment with RDV and high-dose examples 1-6 can significantly improve the increase in brain index (p < 0.001).

[0195] 4.3 Figure 5 A comparative graph showing the changes in lung index (n=6) of OC43-infected suckling mice after treatment in different groups.

[0196] It is evident that viral infection can cause a significant increase in the lung index (p < 0.01), while intervention with RDV (p < 0.05) and high-dose examples 1-6 (p < 0.01) can significantly improve the increase in the lung index.

[0197] 4.4 Figure 6 A comparison of viral load (n=6) in the brain and lung tissues of OC43-infected suckling mice after treatment in different groups.

[0198] It is evident that after viral infection, the viral load in the brain and lung tissue of suckling mice can reach >10. 13 Copy number / g and >10 8 Copy number / g. RDV treatment significantly inhibited viral replication in brain tissue (p < 0.05) and proliferation in lung tissue (p < 0.01). High-dose Examples 1-3 significantly inhibited viral proliferation in brain tissue (p < 0.01) and lung tissue (p < 0.001). High-dose Examples 4-6 significantly inhibited viral replication in brain tissue (p < 0.05) and lung tissue (p < 0.01). Medium-dose Examples 1-5 significantly inhibited viral replication in brain tissue (p < 0.05). Medium-dose Example 6 significantly inhibited viral replication in lung tissue (p < 0.05). Medium-dose Example 6 had no significant inhibitory effect on viral replication in brain tissue. Low-dose Examples 1-6 had no significant inhibitory effect on viral replication in either brain or lung tissue.

[0199] 5. Conclusion

[0200] Appropriate doses of the antiviral Chinese medicine formula granules in Examples 1-6 can improve weight loss in suckling mice infected with coronavirus HCoV-OC43, significantly improve the increase in brain and lung indices caused by viral infection in suckling mice, and show a significant inhibitory effect on viral proliferation in the brain and lung tissues of suckling mice.

[0201] III. Toxicological Studies – Acute Toxicity

[0202] 1. Experimental Materials

[0203] 1.1 Drugs

[0204] Antiviral Traditional Chinese Medicine Formula Granules (Examples 1-6)

[0205] 1.2 Animals

[0206] 120 SPF-grade SD rats, half male and half female, weighing 180-200g, were provided by Southern Medical University.

[0207] 1.3 Feeding conditions

[0208] Five animals were housed in barrier-controlled IVC cages. The facility temperature was 20–26°C, and the humidity was 40–70%. The feed supplier was Co-Bio.

[0209] 2 Experimental Methods

[0210] In accordance with the "Technical Guidelines for Single-Dose Toxicity Studies of Drugs" (Appendix 2 of Announcement No. 4 of 2014 issued by the State Food and Drug Administration), the maximum dose method was adopted. Animals were acclimatized for 5 days, with fasting but water allowed overnight before administration. Administration was performed via oral gavage in three divided doses over 24 hours, with a total dose of 16,000 mg / kg body weight. Animals were observed for 14 days after administration. Observational records included clinical symptoms (such as animal appearance, behavior, diet, response to stimuli, secretions, and excretions), mortality (time of death, pre-mortem reactions, etc.), and weight changes (weighed before administration, at the end of the observation period, and once during the observation period; weighed upon death or near death). After the observation period, the animals were euthanized and subjected to gross necropsy.

[0211] 3 Experimental Results

[0212] No abnormalities or deaths were observed in the animals during the observation period. Gross autopsies at the end of the observation period revealed no obvious lesions. No signs of toxicity were found at this dosage. Details are shown in Tables 6-11. Table 6 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 1; Table 7 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 2; Table 8 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 3; Table 9 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 4; Table 10 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 5; and Table 11 shows the observation results of the antiviral traditional Chinese medicine formula granules administered for 14 days in Example 6.

[0213] Table 6

[0214]

[0215]

[0216] Table 7

[0217]

[0218]

[0219] Table 8

[0220]

[0221] Table 9

[0222]

[0223]

[0224] Table 10

[0225]

[0226] Table 11

[0227]

[0228] IV. Toxicological Studies – Chronic Toxicity

[0229] 1. Experimental Materials

[0230] 1.1 Drugs

[0231] Antiviral Traditional Chinese Medicine Formula Granules of Example 1

[0232] 1.2 Animals

[0233] Eighty SPF-grade SD rats, half male and half female, weighing 150-180g, were provided by Southern Medical University.

[0234] 1.3 Feeding conditions

[0235] Five animals were housed in barrier-controlled IVC cages. The facility temperature was 20–26°C, and the humidity was 40–70%. The feed supplier was Co-Bio.

[0236] 2 Experimental Methods

[0237] (1) Trial grouping and dosage design

[0238] In accordance with the "Technical Guidelines for Repeated-Dose Toxicity Studies of Drugs" (Appendix 3 of Announcement No. 4 of 2014 issued by the State Food and Drug Administration), after 5 days of acclimatization feeding, rats were randomly divided into four groups: a negative control group, a low-dose group, a medium-dose group, and a high-dose group, with 20 rats in each group (half male and half female). Based on body weight data, the drug dosage for the sample groups was: 2000 mg / kg body weight for the low-dose group, 4000 mg / kg body weight for the medium-dose group, and 8000 mg / kg body weight for the high-dose group. The negative control group was given a certain volume of sterile water.

[0239] (2) Observation and Recording

[0240] Observe the animals daily, including but not limited to their physical appearance, behavior, glandular secretions, respiration, and fecal characteristics. Record observations promptly, and promptly dissect any dead or dying animals. Weigh the animals before the start of the experiment, weekly, and before dissection. The drug administration cycle is 28 days, and the weekly dosage volume is adjusted based on the weekly weighing data.

[0241] (3) Anatomical sampling and testing indicators

[0242] After the last administration, the animals were kept on a fasting and water-free overnight. Blood was collected from the abdominal main vein after anesthesia to test blood biochemistry and routine blood indicators. The animals were euthanized and grossly dissected. Five organs were harvested: heart, liver, spleen, kidney, and brain. For males, the testes and epididymis were also harvested, and for females, the uterus and ovaries were also harvested. The organs were weighed and fixed in 10% neutral formalin solution. Histopathological examination can be performed later, and the organ coefficients of each organ are calculated based on the pre-dissection weight.

[0243] 3. Experimental Results

[0244] 3.1 During the experiment, the animals in each group were in good spirits, active, and showed no obvious abnormalities.

[0245] 3.2 The changes in average body weight of animals in each group during the experimental period are shown in Table 12, in g.

[0246] Table 12

[0247]

[0248] 3.3 Blood Routine Indicators: Compared with the same-sex negative control group, there were no statistically significant differences in blood routine indicators among male individuals (P>0.05); the mean erythrocyte volume (MCV) of the high-dose group of female individuals was higher than that of the negative control group, and the difference was statistically significant (P<0.05), but no dose-response relationship was established. Statistical data of blood routine indicators are shown in Table 13 (mean ± standard deviation), except that platelet-weighted hematocrit was too low to be measured.

[0249] Table 13

[0250]

[0251]

[0252] Compared with the negative control group, *p<0.05

[0253] 3.4 Blood biochemical indicators: Compared with the same-sex negative control group, the AST of male individuals in the high-dose group was higher than that of the negative control group, and the difference was statistically significant (P<0.05), but no dose-response relationship was established. The ALP of female individuals in the low-dose and high-dose groups, the T-Bil of the medium-dose group, the BUN of the low-dose and medium-dose groups, and the GLU of the low-dose, medium-dose, and high-dose groups were all higher than those of the negative control group, and the differences were statistically significant (P<0.05), but no dose-response relationship was established. Statistical data of blood biochemical indicators are shown in Table 14 (mean ± standard deviation).

[0254] Table 14

[0255]

[0256]

[0257] Compared with the negative control group, *p<0.05

[0258] 3.5 Gross Anatomy and Organ Coefficients: No obvious lesions were found in the gross anatomical results. Compared with the negative control group of the same sex, there was no statistically significant difference in organ coefficients among male individuals (P>0.05). The liver index of female individuals in the high-dose group was higher than that in the negative control group, and the difference was statistically significant (P<0.05), but no dose-response relationship was established. The mean organ coefficients (wet weight of organs / body weight before dissection*100%) of each group are shown in Table 15 (mean ± standard deviation, %).

[0259] Table 15

[0260]

[0261]

[0262] Compared with the negative control group, *p<0.05

[0263] 4. Conclusion

[0264] One-way ANOVA was used to analyze blood routine and blood biochemistry data. The results showed significant differences in some indicators, but no dose-response relationship was established, and there were no significant differences in organ indices. No obvious toxic reactions or target organs were found at any dose in this experiment.

[0265] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0266] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A traditional Chinese medicine composition for treating coronaviruses, characterized in that, It is made from the following components in parts by weight: Magnolia officinalis 8-18 parts, Trichosanthes kirilowii 10-20 parts, Scutellaria baicalensis 5-15 parts, Bupleurum chinense 5-15 parts, Atractylodes lancea 2-10 parts, Zingiber officinale 2-10 parts, Adenophora stricta 2-10 parts, and Paeonia lactiflora 3-11 parts.

2. The anticoronavirus traditional Chinese medicine composition according to claim 1, characterized in that, It is made from the following components in parts by weight: Magnolia officinalis 10-15 parts, Trichosanthes kirilowii 13-18 parts, Scutellaria baicalensis 8-13 parts, Bupleurum chinense 8-13 parts, Atractylodes lancea 3-8 parts, fresh ginger 3-8 parts, Adenophora stricta 3-8 parts, and Paeonia lactiflora 4-9 parts.

3. The anticoronavirus traditional Chinese medicine composition according to claim 1 or 2, characterized in that, It is made from the following components in parts by weight: Magnolia officinalis 12-13 parts, Trichosanthes kirilowii 15-16 parts, Scutellaria baicalensis 10-11 parts, Bupleurum chinense 10-11 parts, Atractylodes lancea 5-6 parts, fresh ginger 5-6 parts, Adenophora stricta 5-6 parts, and Paeonia lactiflora 6-7 parts.

4. A traditional Chinese medicine formula granule for treating coronavirus, characterized in that, It is made from the anticoronavirus traditional Chinese medicine composition according to any one of claims 1 to 3 and pharmaceutically acceptable excipients.

5. The anti-coronavirus traditional Chinese medicine formula granules according to claim 4, characterized in that, The pharmaceutically acceptable excipients include one or more of dextrin, beta-cyclodextrin, lactose, silicon dioxide, and magnesium stearate.

6. A method for preparing a traditional Chinese medicine formula granule for treating coronavirus, characterized in that, Includes the following steps: Take the medicinal materials of each component as described in any one of claims 1 to 3, and prepare formulation granules of each component; The formulation granules of each component are mixed to prepare anti-coronavirus traditional Chinese medicine formulation granules.

7. The method for preparing anti-coronavirus traditional Chinese medicine formula granules according to claim 6, characterized in that, The method for preparing formulation particles of each component includes the following steps: Take the medicinal materials of each component, add water to extract them separately, and obtain the extracts of each component; The extracts of each component were concentrated to obtain concentrated solutions of each component. Pharmaceutically acceptable excipients were added to the concentrates of each component, and the mixture was spray-dried to obtain the formulation granules of each component.

8. The method for preparing anti-coronavirus traditional Chinese medicine formula granules according to claim 7, characterized in that, The mass of the water is 3 to 8 times the mass of the medicinal material to be extracted; and / or The extraction is performed 2-3 times, and the extracts from each extraction are combined to obtain a single-component extract; and / or The relative density of the concentrated solutions of each component is 1.05~1.

20.

9. The use of the anti-coronavirus traditional Chinese medicine composition according to any one of claims 1 to 3 or the anti-coronavirus traditional Chinese medicine formula granules according to any one of claims 4 to 5 in the preparation of anti-coronavirus HCoV-OC43 drugs.

10. The use of the anti-coronavirus traditional Chinese medicine composition according to any one of claims 1 to 3 or the anti-coronavirus traditional Chinese medicine formula granules according to any one of claims 4 to 5 in the preparation of drugs against the novel coronavirus SARS-CoV-2.

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