FORSYTHIA SUSPENSA COMPONENT AND OPTIONAL PANAX GINSENG COMPONENT AND THE APPLICATION

Abstract

In one aspect, the present invention discloses a composition of forsythia suspensa component and an optional panax ginseng component that can be used safely, efficiently and consistently to achieve antiviral, antipyretic, anti-inflammatory and/or immunity boosting effects, including forsythin, forsythin derivatives, and the composition of forsythin and phillygenin for the prevention and treatment of diseases associated with human cytomegalovirus infection, such as simple human cytomegalovirus infection or for patients infected by human cytomegalovirus during prevention and treatment of cardiovascular and cerebrovascular diseases, tumors, bums or AIDS or during a process of organ transplantation or during the perinatal period. In another aspect, this application provides a process for preparing the composition of forsythia suspensa and panax ginseng components is simple, suitable for industrial production and easy to popularize.

Claims

1. A composition for achieving the antiviral, antipyretic, anti-inflammatory and/or immunity boosting effects, which includes the forsythia suspensa components and preferably the panax ginseng components.

2. The composition of claim 1, wherein the panax ginseng components comprise the medicinal material of panax ginseng, the extract of panax ginseng, total ginsenoside, panaxadiol saponins, ginsenoside Rg3 or 20 (R)-ginsenoside Rg3; and the forsythia suspensa components include the medicinal material of forsythia suspensa leaf, the extract of forsythia suspensa leaf, forsythin and phillygenin and/or forsythin derivatives.

3. The composition of claim 1, wherein the extract is an alcoholic or aqueous extract.

4. The composition of claim 1, wherein the weight ratio of panax ginseng components to forsythia suspensa components is 2-98: 98-2.

5. The composition of claim 1, wherein the composition further includes one or more of the following components: panax notoginseng, aloe vera, glycyrrhiza uralensis, fallopia multiflora, ginkgo biloba, black sesame extract, ginger extract, grape seed extract, pomegranate seed extract, plant essential oil, arbutin, vitamin C and its derivatives or vitamin E and its derivatives; or, it also includes one or more of the following components: sophora tonkinensis extract, xanthium sibiricum extract, scutellaria barbata extract, Sophora flavescens extract, dandelion extract, honeysuckle extract, zingiber officinale rhizome extract, grape seed extract, pomegranate seed extract, vitamin C and its derivatives or vitamin E and its derivatives.

6. The composition of claim 1, wherein the composition does not include a panax ginseng component, wherein the forsythia component is forsythin and its derivatives, or the composition of forsythin and phillygenin.

7. The composition of claim 6, wherein the forsythin derivatives include 33-hydroxy phillygenin-8-O-β-D-glucuronide, 9-hydroxy phillygenin-8-O-β-D-glucuronide, 33, 34-methylenedioxy phillygenin-8-O-β-D-glucuronide, (2R,3R,4R,5S)-methyl 6-(5-((1R,4S)-4-(3,4-dimethoxyphenyl) hexahydrofuro[3,4-c] furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxyte trahydro-2H-pyran-2-carboxylate, sodium (2R,3R,4R,5S)-6-(5- ((1R,4S) -4- (3,4-dimethoxyphenyl) hexahydrofuro [3,4-c]furan-1-yl) -2- methoxyphenoxy)-3,4,5-trihydroxytetrahydro -2H -pyran-2-carboxylate, potassium (2R,3R,4R,5S)-6- (5-((1R,4S)-4- (3,4-dimethoxyphenyl) hexahydrofuro[3,4-c]furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro- 2H-pyran-2-carboxylate or (2R,3R, 4R,5S)-6-(5-((1R,4S)-4-(3,4-dimethoxyphenyl) hexahydrofuro[3,4-c]furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid; preferably 33-hydroxy phillygenin-8-O-β-D-glucuronide, 9-hydroxy phillygenin-8-O-β-D-glucuronide, 33, 34-methylenedioxy phillygenin-8-O-β-D-glucuronide, (2R,3R,4R,5S)-methyl 6-(5-((1R,4S)-4-(3,4-dimethoxyphenyl) hexahydrofuro[3,4-c] furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxyte trahydro-2H-pyran-2-carboxylate, sodium (2R,3R,4R,5S)-6-(5- ((1R,4S) -4- (3,4-dimethoxyphenyl) hexahydrofuro [3,4-c]furan-1-yl) -2- methoxyphenoxy)-3,4,5-trihydroxytetrahydro -2H -pyran-2-carboxylate and potassium (2R,3R,4R,5S)-6- (5-((1R,4S)-4- (3,4-dimethoxyphenyl) hexahydrofuro[3,4-c]furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro- 2H- pyran-2-carboxylate.

8. The composition of claim 6, wherein the composition of forsythin and phillygenin consists of forsythin and phillygenin, the weight ratio of forsythin to phillygenin is 2-99.99: 0.01-98, preferably at 80-99:1-20.

9. A preparation for achieving the antiviral, antipyretic, anti-inflammatory and/or immunity-boosting effects, wherein the preparation comprises the composition of claim 1 and excipients that are acceptable in the industries of pharmaceutics, health products or foods.

10. The preparation of claim 9, wherein the weight ratio of the composition and the carrier acceptable in the industries of pharmaceutics, health products or foods is 1:1 to 1:100, or the weight ratio is 0.01-50:100.

11. The preparation of claim 9, wherein the carrier acceptable in the industries of pharmaceutics, health products or foods is a cyclodextrin, such as α-, β-or γ-cyclodextrin or its derivatives.

12. The preparation of claim 9, wherein the preparation is a drug product, which may exist in the preferable dosage forms of tablets, capsules, pills, powder, granules, syrup, solution, emulsion, injection, spray, aerosol, gel, cream, cataplasm, adhesive plaster or emplastrum.

13. The preparation of claim 9, wherein the preparation is a health product or food, which may exist in the preferable dosage forms of tablets, capsules, pills, powder, granules, syrup, solution, emulsion, spray, aerosol, gel, cream, cataplasm, adhesive plaster or emplastrums, or in the forms of foods such as dairy products, confectionery, beverages, biscuits, tea leaves and related products, wine and the like.

14. A method for preparing the composition of claim 1, wherein the method comprises a step of blending the panax ginseng component and forsythia suspensa component.

15. A method for preparing the composition of claim 1, wherein the method comprises a step of using the solvent heating extraction method to extract the panax ginseng and forsythia suspensa and optional Chinese medicinal materials.

16. A method for preparing the preparation of claim 9, wherein the method comprises a step of blending the composition and a carrier acceptable in the industries of pharmaceutics, health products or foods.

17. The method of claim 16, wherein the method comprises a step of blending the composition with a cyclodextrin or its derivative, or α-, β-or γ-cyclodextrin or a derivative thereof.

18. The method of claim 16, wherein the method comprises a step of using physical or chemical methods to treat the composition with a cyclodextrin or a derivative thereof, or with α-, β-or γ-cyclodextrin or a derivative thereof to form a complex.

19. The use of the composition of claim 1 in manufacturing drugs, health products or foods for achieving the antiviral, antipyretic, anti-inflammatory and/or immunity-boosting effects.

20. A method for achieving antiviral, antipyretic, anti-inflammatory and/or immunity-boosting effects, wherein the method comprises administering an effective amount of the composition of claim 1 to an individual in need of achieving the antiviral, antipyretic, anti-inflammatory and/or immunity-boosting effects.

21. The use of the composition of claim 6 in the manufacturing of drugs, health products or foods for the anti-cytomegalovirus effects or the prevention or treatment of disorders caused by cytomegalovirus infection.

22. A method for achieving an anti-cytomegalovirus effect or for prevention or treatment of disorders caused by cytomegalovirus infection, wherein the method comprises administering an effective amount of the composition of claim 6 to an individual who needs the composition.

23. The method of claim 22, wherein the cytomegalovirus is human cytomegalovirus.

24. The method of claim 22, wherein the condition comprises a simple human cytomegalovirus infection condition, such as simple human cytomegalovirus infection or patients infected by human cytomegalovirus during the treatment of cardiovascular and cerebrovascular diseases, tumors, burns or AIDS or during the process of organ transplantation or during the perinatal period.

Description

DETAILED DESCRIPTION

[0140] The beneficial effects of the formulations described in the present invention are further described below using the specific embodiments. These exemplary embodiments are exemplary only and do not constitute any limitation to the scope of the present invention. It should acknowledged by those skilled in tile art that the details and forms of the technical proposal of the present invention can be modified or substituted without departing from the formulation ideas and scope of use of the present invention. However, all these modifications and substitutions fall within the scope of protection of the present invention.

[0141] The said forsythin derivatives 33-hydroxy-phillygenin glucuronide, 9-hydroxy-phillygenin glucuronide and 33, 34-methylene dioxy-phillygenin glucuronide in the present invention are the same as the forsythin derivatives described in the patent application (application No.: 201510319303.4, priority No.: 201510164294.6). The (2R,3R,4R,5S)-methyl 6-(5-((1R,4S)-4-(3,4-dimethoxyphenyl) hexahydrofuro[3.4-c] furan-1-yl)-2-methoxyphenoxy)-3.4.5-trihydroxyte trahydro-2H-pyran-2-carboxylate, sodium (2R,3R,4R.5S)-6-(5- ((1R,4S) -4- (3,4-dimethoxyphenyl) hexahydrofuro [3,4-c]furan-1-yl) -2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro -2H - pyran-2-carboxylate and potassium (2R,3R,4R,5S)-6-(5-((1R,4S)-4- (3,4-dimethoxyphenyl) hexahydrofuro[3,4-c]furan-1-yl)-2-methoxyphenoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylate and forsythione glucuronic acid are the same as those described in the patent application (application No.: 201510320579.4. priority No.: 201410825547.5).

Exemplary Embodiment 1: Forsythin Tablets

[0142] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00001 Forsythin (purity: 86%) 500 g Starch 1000 g Talc 15 g Magnesium stearate 15 g

[0143] 2. Mix forsythin with starch by the above ratios, mix well and prepare the mixture into granules, add talc and magnesium stearate, mix well, and then compress the mixture into 10,000 tablets.

Exemplary Embodiment 2: Forsythin Capsules

[0144] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00002 Forsythin (purity: 60%) 200 g Starch 1000 g

[0145] 2. Mix forsythin with starch, mix well and dispense it into 10,000 capsules.

Exemplary Embodiment 3: Forsythin Capsules

[0146] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00003 Forsythin (purity: 98%) 500 g Starch 1000 g

[0147] 2. Mix forsythin with starch, mix well and dispense it into 10,000 capsules.

Exemplary Embodiment 4: Forsythin Tablets

[0148] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00004 Forsythin (purity: 63%) 50 g Evodia spp. fruit extract 200 g Vitamin C 400 g Starch 1000 g Talc 16.5 g Magnesium stearate 16.5 g

[0149] 2. Mix forsythin, evodia spp. fruit extract vitamine C and starch, mix well and prepare the mixture into granules, then add talc and magnesium stearate, mix well, and then compress the mixture into 10,000 tablets.

Exemplary Embodiment 5: Forsythin Capsules

[0150] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00005 Forsythin (purity: 98%) 300 g Alpinia officinarum extract 300 g Vitamin C 600 g Starch 1000 g

[0151] 2. Mix forsythin, alpinia officinarum extract, vitamine C and starch, mix well and dispense it into 10,000 capsules.

Exemplary Embodiment 6: Forsythin Granules

[0152] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00006 Forsythin (purity: 92%) 500 g Amomi fructus extract 30 g Vitamin C 600 g Sucrose powder 5000 g

[0153] 2. Mix forsythin, amomi fructus extract, vitamine C and sucrose powder, mix well and prepare it into granules and dispense it into 10,000 sachets.

Exemplary Embodiment 7: Forsythin Oral Solution

[0154] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00007 Forsythin (purity: 98%) 4 g Bamboo shavings extract 3 g Phragmitis rhizoma extract 3 g Glucose syrup 5 g Deionized water Appropriate amount

[0155] Take forsythin, bamboo shavings extract and phragmitis rhizoma extract, dissolve with ethanol, add glucose syrup, then add deionized water to 100 ml, and mix well to obtain the product.

Exemplary Embodiment 8: 33-Hydroxy-phillygenin Glucuronide Tablets

[0156] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00008 33-hydroxy-phillygenin glucuronide (purity 66%) 500 g Starch 1000 g Talc 15 g Magnesium stearate 15 g

[0157] 2. Mix 33-hydroxy-phillygenin glucuronide and starch well, granulate, add talc and magnesium stearate, and compress to make 10000 tablets.

Exemplary Embodiment 9: 33-Hydroxy-phillygenin Glucuronide Capsules

[0158] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00009 33-hydroxy-phillygenin glucuronide (purity 60%) 200 g Starch 1000 g

[0159] 2. Mix 33-hydroxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 10: 33-Hydroxy-phillygenin Glucuronide Capsules

[0160] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00010 33-hydroxy-phillygenin glucuronide (purity 98%) 500 g Starch 1000 g

[0161] 2. Mix 33-hydroxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 11: 33-Hydroxy-phillygenin Glucuronide Tablets

[0162] 1. Prepare the raw materials according to the following ratios:

TABLE-US-00011 33-hydroxy-phillygenin glucuronide (purity 63%) 50 g Evodia spp. fruit extract 20 g Vitamin C 400 g Starch 1000 g Talc 14.7 g Magnesium stearate 14.7 g

[0163] 2. Mix 33-hydroxy-phillygenin glucuronide, evodia spp. fruit extract, vitamin C and starch well, granulate, add talc and magnesium stearate and mix well and compress to make 10000 tablets.

Exemplary Embodiment 12: 33-Hydroxy-phillygenin Glucuronide Capsules

[0164] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00012 33-Hydroxy-phillygenin glucuronide (purity 98%) 300 g Pinellia ternata extract 30 g Vitamin C 600 g Starch 1000 g

[0165] 2. Mix 33-hydroxy-phillygenin glucuronide, pinellia ternata extract, vitamin C and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 13: 33-Hydroxy-phillygenin Glucuronide Granules

[0166] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00013 33-Hydroxy-phillygenin glucuronide (purity 92%) 500 g Amomum cardamomum extract 30 g Vitamin C 600 g Sucrose powder 5000 g

[0167] 2. Mix 33-hydroxy-phillygenin glucuronide, amomum cardamomum extract, vitamin C and sucrose powder well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 14: 33-Hydroxy-phillygenin Glucuronide Oral Solution

[0168] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00014 33-Hydroxy-phillygenin glucuronide (purity 98%) 4 g Evodia spp. fruit extract 3 g Amomum cardamomum extract 3 g Glucose syrup 5 g Deionized water Appropriate amount

[0169] 2. Dissolve 33-hydroxy-phillygenin glucuronide, amomum cardamomum extract and evodia spp. fruit extract in ethanol, add glucose syrup, and finally add deionized water to 100 mL.

Exemplary Embodiment 15: 9-Hydroxy-phillygenin Glucuronide Tablets

[0170] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00015 9-Hydroxy-phillygenin glucuronide (purity 66%) 500 g Starch 1000 g Talc 15 g Magnesium stearate 15 g

[0171] 2. Mix 9-hydroxy-phillygenin glucuronide and starch well, granulate, add talc and magnesium stearate, and compress to make 10000 tablets.

Exemplary Embodiment 16: 9-Hydroxy-phillygenin Glucuronide Capsules

[0172] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00016 9-Hydroxy-phillygenin glucuronide (purity 97%) 200 g Starch 1000 g

[0173] 2. Mix 9-hydroxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 17: 9-Hydroxy-phillygenin Glucuronide Capsules

[0174] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00017 9-Hydroxy-phillygenin glucuronide (purity 98%) 500 g Starch 1000 g

[0175] 2. Mix 9-hydroxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 18: 9-Hydroxy-phillygenin Glucuronide Tablets

[0176] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00018 9-Hydroxy-phillygenin glucuronide (purity 63%) 50 g Evodia spp, fruit extract 20 g Vitamin C 400 g Starch 1000 g Talc 14.7 g Magnesium stearate 14.7 g

[0177] 2. Mix 9-hydroxy-phillygenin glucuronide, evodia spp. fruit extract, vitamin C and starch, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 19: 9-Hydroxy-phillygenin Glucuronide Capsules

[0178] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00019 9-Hydroxy-phillygenin glucuronide (purity 98%) 300 g Pinellia ternata extract 30 g Vitamin C 600 g Starch 1000 g

[0179] 2. Mix 9-hydroxy-phillygenin glucuronide, pinellia ternata extract, vitamin C and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 20: 9-Hydroxy-phillygenin Glucuronide Granules

[0180] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00020 9-Hydroxy-phillygenin glucuronide (purity 92%) 500 g Amomum cardamomum extract 30 g Vitamin C 600 g Sucrose powder 5000 g

[0181] 2. Mix 9-hydroxy-phillygenin glucuronide, amomum cardamomum extract, vitamin C and sucrose powder well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 21: 9-Hydroxy-phillygenin Glucuronide Oral Solution

[0182] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00021 9-Hydroxy-phillygenin glucuronide (purity 98%) 4 g Inula japonica inflorescence extract 3 g Amomum cardamomum extract 3 g Glucose syrup 5 g Deionized water Appropriate amount

[0183] 2. Dissolve 9-hydroxy-phillygenin glucuronide, amomum cardamomum extract and inula japonica inflorescence extract in ethanol, add glucose syrup, and finally add deionized water to 100 mL.

Exemplary Embodiment 22: 33,34-Methylenedioxy-phillygenin Glucuronide Tablets

[0184] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00022 33,34-Methylenedioxy-phillygenin glucuronide (purity 66%) 500 g Starch 1000 g Talc 15 g Magnesium stearate 15 g

[0185] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide and starch well, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 23: 33,34-Methylenedioxy-phillygenin Glucuronide Capsules

[0186] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00023 33,34-Methylenedioxy-phillygenin glucuronide (purity 97%) 200 g Starch 1000 g

[0187] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 24: 33,34-Methylenedioxy-phillygenin Glucuronide Capsules

[0188] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00024 33,34-Methylenedioxy-phillygenin glucuronide (purity 98%) 500 g Starch 1000 g

[0189] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 25: 33,34-Methylenedioxy-phillygenin Glucuronide Tablets

[0190] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00025 33,34-Methylenedioxy-phillygenin glucuronide (purity 63%) 50 g Inula japonica inflorescence extract 20 g Vitamin C 400 g Starch 1000 g Talc 14.7 g Magnesium stearate 14.7 g

[0191] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide, inula japonica inflorescence extract, vitamin C and starch well, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 26: 33,34-Methylenedioxy-phillygenin Glucuronide Capsules

[0192] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00026 33.34-Methylenedioxy-phillygenin glucuronide (purity 98%) 300 g Pinellia ternata extract 30 g Vitamin C 600 g Starch 1000 g

[0193] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide, pinellia extract, vitamin C and starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 27: 33,34-Methylenedioxy-phillygenin Glucuronide Granules

[0194] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00027 33,34-Methylenedioxy-phillygenin glucuronide (purity 92%) 500 g Amomum cardamomum extract 30 g Vitamin C 600 g Sucrose powder 5000 g

[0195] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide, amomum cardamomum extract, vitamin C and sucrose powder well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 28: 33,34-Methylenedioxy-phillygenin Glucuronide Granules

[0196] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00028 33,34-Methylenedioxy-phillygenin glucuronide (purity 98%) 300 g Amomum cardamomum extract 30 g Vitamin C 600 g Sucrose powder 5000 g Make 10000 bags

[0197] 2. Mix 33,34-methylenedioxy-phillygenin glucuronide, amomum cardamomum extract, vitamin C and sucrose powder well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 29: 33,34-Methylenedioxy-phillygenin Glucuronide Oral Solution

[0198] 1. Prepare the raw materials according to the following ratio:

TABLE-US-00029 33.34-Methylenedioxy-phillygenin glucuronide (purity 98%) 4 g Inula japonica inflorescence extract 3 g Amomum cardamomum extract 3 g Glucose syrup 5 g Deionized water Appropriate amount

[0199] 2. Dissolve 33,34-methylenedioxy-phillygenin glucuronide, amomum cardamomum extract and inula japonica inflorescence extract in ethanol, add glucose syrup, and finally add deionized water to 100 mL.

Exemplary Embodiment 30: Preparation of Forsythin and Phillygenin Composition Tablets

[0200] 1. Prepare forsythin/phillygenin composition tablets according to the following ratio:

TABLE-US-00030 Forsythin/phillygenin composition (weight ratio of 98: 2) 500 g Starch 480 g Talc 10 g Magnesium stearate 10 g

[0201] 2. Mix 490 g of forsythin and 10 g of phillygenin with starch well, granulate, add talc and magnesium stearate, and compress to make 10000 tablets.

Exemplary Embodiment 31: Preparation of Forsythin/Phillygenin Composition Granules

[0202] 1. Prepare forsythin/phillygenin composition granules according to the following ratio:

TABLE-US-00031 Forsythin/phillygenin composition (weight ratio of 98: 2) 100 g Microcrystalline cellulose 10000 g

[0203] 2. Mix 98 g of forsythin and 2 g of phillygenin with microcrystalline cellulose well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 32: Preparation of Forsythin/Phillygenin Composition Capsules

[0204] 1. Prepare forsythin/phillygenin composition capsules according to the following ratio:

TABLE-US-00032 Forsythin/phillygenin composition (weight ratio of 98: 2) 250 g Starch 2500 g

[0205] 2. Mix 245 g of forsythin and 5 g of phillygenin with starch well, and fill it into capsules to make 10000 capsules.

Exemplary Embodiments 33-36: Preparation of Forsythin/Phillygenin Composition Capsules

[0206] In Exemplary Embodiments 33-36. the forsythin/phillygenin composition is mixed with starch in the weight ratios in the following table and then filled into capsules to make 10000 capsules, respectively.

TABLE-US-00033 Exemplary Embodiment No Raw material Phillygenin and forsythin composition/g Excipient Starch/g Weight ratio of raw material to excipient Exemplary Embodiment 33 500(98:2) 500 1:1 Exemplary Embodiment 34 50 (80:20) 5000 1:100 Exemplary Embodiment 35 250(90:10) 2500 1:10 Exemplary Embodiment 36 250(95:5) 5000 1:20

Exemplary Embodiments 37-40: Preparation of Forsythin/Phillygenin Composition Granules

[0207] In Exemplary Embodiments 37-40, the forsythin/phillygenin composition is mixed with microcrystalline cellulose according to the weight ratios in the following table, granulated and packed into 10000 bags.

TABLE-US-00034 Exemplary Embodiment No Raw material Phillygenin and forsythin composition/g Excipient Microcrystalline cellulose/g Weight ratio of raw material to excipient Exemplary Embodiment 37 1000(98:2) 1000 1:1 Exemplary Embodiment 38 250 (80:20) 25000 1:100 Exemplary Embodiment 39 2500(90:10) 25000 1:10 Exemplary Embodiment 30 2500(95:5) 50000 1:20

Exemplary Embodiment 41: Preparation of Forsythin/Phillygenin Composition Tablets

[0208] 1. Prepare forsythin/phillygenin composition tablets according to the following ratio:

TABLE-US-00035 Forsythin/phillygenin composition (weight ratio of 98: 2) 500 g Starch 380 g Agastache rugosa extract 100 g Talc 10 g Magnesium stearate 10 g

[0209] 2. Mix 490 g of forsythin and 10 g of forsythin with agastache rugosa extract powder well, then mix well with starch, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 42: Preparation of Forsythin/Phillygenin Composition Granules

[0210] 1. Prepare forsythin/phillygenin composition granules according to the following ratio:

TABLE-US-00036 Fonsythin/phillygenin composition (weight ratio of 98: 2) 250 g Aquilaria sinensis extract 250 g Agastache rugosa extract 250 g Microcrystalline cellulose 24500 g

[0211] 2. Mix 245 g of forsythin and 5 g of phillygenin with aquilaria sinensis extract and agastache rugosa extract powder, then mix well with microcrystalline cellulose, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 43: Preparation of Forsythin/Phillygenin Composition Capsules

[0212] 1. Prepare forsythin/philly genin composition capsules according to the following ratio:

TABLE-US-00037 Forsythin/philly genin composition (weight ratio of 98: 2) 250 g Perilla frutescens extract 250 g Eriobotrya japonica leaf extract 250 g Syzygium aromaticum extract 250 g Starch 1000 g

[0213] 2. Mix 245 g of forsythin and 5 g of phillygenin, perilla frutescens extract, eriobotrya japonica leaf extract and syzygium aromaticum extract powder well, then mix well with starch, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 44: Preparation of Forsythin/Phillygenin Composition Tablets

[0214] 1. Prepare forsythin/phillygenin composition tablets according to the following ratio:

TABLE-US-00038 Forsythin/phillygenin composition (weight ratio of 80: 20) 500 g Starch 480 g Eriobotrya japonica leaf extract 500 g Talc 10 g Magnesium stearate 10 g

[0215] 2. Mix 490 g of forsythin and 10 g of phillygenin with eriobotrya japonica leaf extract powder well, then mix well with starch, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 45: Preparation of Forsythin/Phillygenin Composition Granules

[0216] 1. Prepare forsythin/phillygenin composition granules according to the following ratio:

TABLE-US-00039 Forsythin/phillygenin composition (weight ratio of 90: 10) 1000 g Inula japonica inflorescence extract 500 g Pinellia ternata extract 500 g Microcrystalline cellulose 10000 g

[0217] 2. Mix 900 g of forsythin and 100 g of phillygenin with the above extracts (inula japonica inflorescence, pinellia ternata), then mix well with microcrystalline cellulose, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 46: Preparation of Forsythin/Phillygenin Composition Capsules

[0218] 1. Prepare forsythin/phillygenin composition capsules according to the following ratio:

TABLE-US-00040 Forsythin/philly genin composition (weight ratio of 94: 6) 2000 g Inula japonica inflorescence extract 250 g Katsumade galangal seed extract 500 g Eriobotrya japonica leaf extract 250 g Starch 1000 g

[0219] 2. Mix 1880 g of forsythin and 120 g of phillygenin with the above extracts (inula japonica inflorescence, katsumade galangal seed, eriobotrya japonica leaf) well, then mix well with starch, and fill it into capsules to make 10000 capsules.

Exemplary Embodiment 47: Preparation of Forsythin/Phillygenin Composition Soft Capsules

[0220] 1. Prepare forsythin/phillygenin composition soft capsules according to the following ratio:

TABLE-US-00041 Forsythin/phillygenin composition (weight ratio of 94: 6) 2000 g β-cyclodextrin 1500 g PEG-400 2000 g PEG-4000 26 g Propylene glycol 260 g

[0221] 2. Heat 2000 g of PEG-400, 26 g of PEG-4000 and 260 g of propylene glycol to 65° C. to melt mix well, and use this as a water-soluble matrix: Mix with the composition and mix well. Take 5000 g of gelatin and 4500 g of water, heat to 70° C. take 1750 g of glycerin and 15 g of methylparaben, stir, evacuate for 1 h, mix with the above gelatin, maintain at 70° C. and allow to stand overnight. Prepare with an automatic soft capsule filling machine to obtain 10000 soft capsules.

Exemplary Embodiment 48: Experimental Study of Forsythin, Forsythin Derivative, Forsythin and Phillygenin Composition against Human Cytomegalovirus in Vitro

1. Experimental Materials

1.1 Drugs and Reagents

[0222] Forsythin (Batch No.: 20130303), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co.. Ltd. The purity was determined to be 99.5% by high performance liquid chromatography (HPLC). The content was confirmed to be 99.5% by calibration with the forsythin reference standard for assay of pharmaceutical and biological products in China.

[0223] 33-Hydroxy-forsythin glucuronide (forsythin derivative A. batch No.: 20130301), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. The purity was determined to be 98.5% by area normalization with two detectors (ultraviolet detector and evaporative light scattering detector) of high performance liquid chromatography.

[0224] 9-Hydroxy-forsythin glucuronide (forsythin derivative B, batch number: 20130302), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co.. Ltd. The purity was determined to be 99.2% by area normalization with two detectors (ultraviolet detector and evaporative light scattering detector) of high performance liquid chromatography.

[0225] 33.34-Methylenedioxy-forsythin glucuronide (forsythin derivative C, batch number: 20130301), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. The purity was determined to be 98.7% by area normalization with two detectors (ultraviolet detector and evaporative light scattering detector) of high performance liquid chromatography.

[0226] Forsythin/phillygenin composition A (Batch No.: 20130305), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co.. Ltd. Upon the content test by high performance liquid chromatography, the ratio of forsythin to phillygenin was 98: 2, and the sum of the contents of forsythin and philly genin was 98.4%. Forsythin/philly genin composition B (Batch No.: 20130306), white powder, manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. Upon the content test by high performance liquid chromatography, the ratio of forsythin to phillygenin was 90: 10. and the sum of the contents of forsythin and phillygenin was 98.1%.

[0227] Ganciclovir (GCV) Powder for Injection (trade name: Li ke wei, manufacturer: Hubei Keyi Pharmaceutical Co., Ltd., batch No.: 091102)

1.2 Viruses and Cells

[0228] Human embryonic lung fibroblast cell strain (purchased from Sino Best Biological Technology Co., Ltd.). The cells were cultured and subcultured according to the conventional method.

[0229] HCMV AD169 strain was purchased from American Type Culture Collection (ATCC).

2. Method

2.1 Viral Virulence Assay

[0230] The cells and viruses were cultured, subcultured and proliferated according to the conventional method, and the TCD.sub.50 of HCMV AD169 strain was 10.sup.-4.6.

2.2. Drug Toxicity Testing

[0231] MTT and cytopathic effect methods were used to determine the maximum non-toxic concentration (TD.sub.0) of each drug and GCV, respectively. The results by the two methods were consistent. The results are shown in Table 1.

2.3. Inhibitory Effect on HCMV

[0232] Cells were inoculated in a 96-well plate, allowed to grow into monolayers, and inoculated with a virus challenge of 100 times TCD.sub.50. Starting from TD.sub.0, each drug was diluted stepwise according to 10-fold gradient to prepare 5 concentrations, with 4 wells set for each concentration. In addition, 4 wells of normal cell control and 4 wells of virus control were set. After 10 days of culture, the minimum effective concentration (MEC) of the drug was investigated, and the therapeutic index (TD.sub.0/MEC) was calculated. The specific data are shown in the table below.

TABLE-US-00042 Summary of Therapeutic Index for Drug and Control Drug name Maximum non-toxic concentration TD.sub.0 Minimum effective concentration MEC Therapeutic index (TD.sub.0/MEC) Ganciclovir (GCV) 10 mg.L.sup.-1 10 mg.L.sup.-1 1 Forsythin 40 mg.L.sup.-1 4 mg.L.sup.-1 10 33-Hydroxy-phillygenin glucuronide 30 mg.L.sup.-1 0.3 mg.L.sup.-1 100 9-Hydroxy-phillygenin glucuronide 40 mg.L.sup.-1 0.4 mg.L.sup.-1 100 33,34-Methylenedioxy-phillygenin glucuronide 40 mg.L.sup.-1 0.4 mg.L.sup.-1 100 Forsythin/philly genin composition A 30 mg.L.sup.-1 3 mg.L.sup.-1 10 Forsythin/philly genin composition B 30 mg.L.sup.-1 3 mg.L.sup.-1 10

3 Discussions

[0233] The inhibitory effects of forsythin, forsythin derivative, forsythin and phillygenin composition on human cytomegalovirus (HCMV) in vitro were investigated using the cytopathic effect method and MTT method with antiviral drug ganciclovir as control. The results showed that forsythin, forsythin derivative, forsythin and phillygenin composition strongly inhibited human cytomegalovirus in vitro, with lower toxicity and higher therapeutic index than those of the control drug. It is suggested that forsythin, forsythin derivative, forsythin and phillygenin composition have potential clinical application prospects for the inhibition of cytomegalovirus.

Exemplary Embodiments 49-52: Preparation of Panax Ginseng-Forsythia Suspensa Composition (Original Powder of Crude Drug)

[0234] Two crude drug powders of panax ginseng and forsythia suspensa are taken, and the two can be mixed according to the weight ratio in Table 2 to prepare the composition of panax ginseng and forsythia suspensa. Panax ginseng and forsythia suspensa are crude drugs meeting the standards of Chinese Pharmacopoeia.

TABLE-US-00043 Raw Material Ratio Table of the Panax Ginseng and Forsythia Suspensa Composition of Exemplary Embodiments 49-52 Exemplary Embodiments No. Weight ratio Panax ginseng Forsythia suspensa (leaf) Exemplary Embodiment 49 98 2 Exemplary Embodiment 50 50 50 Exemplary Embodiment 51 25 75 Exemplary Embodiment 52 2 98

Exemplary Embodiment 53: Preparation of Alcohol Extract of Panax Ginseng-Forsythia Suspensa Composition

1. First Extraction Treatment

[0235] Add pulverized panax ginseng (300 g) and forsythia suspensa (300 g) into a multi-functional extraction tank of traditional Chinese medicine, respectively, mix them with ethanol solution (3000 g) at the volume percentage concentration of 70%, soak for 30 min, turn on the power of the multi-functional extraction tank, heat, perform the first heating extraction treatment on panax ginseng and forsythia suspensa respectively, keep the temperature at 70° C. for constant temperature extraction for 1 h, and then filter to obtain the first extract and the first residue; The weight ratio of panax ginseng and forsythia suspensa separately to the extracting solvent (ethanol solution) is 1:10.

[0236] In the present invention, in addition to the weight ratio of panax ginseng and forsythia suspensa separately to the extracting solvent of 1:10, and other ratios, such as 1:8-12, are applicable to the present invention. In addition to the soaking time of 30 min. other soaking times of ≥ 30 min are applicable to the present invention, preferably 30-60 min: In addition to the ethanol solution at the volume percentage concentration of 70%, other ethanol solutions with a volume percentage concentration of ≥ 50% are applicable to the invention, preferably the ethanol solution at a concentration of 50-75% and absolute ethanol. The extraction temperature of 60-80° C. and the heating extraction time of 0.5-2 h are applicable to the invent.

2. Second Extraction Treatment

[0237] Adding ethanol solution at the volume percentage concentration of 70% to the first residue, heat, perform the second heating extraction treatment on panax ginseng and forsythia suspensa, respectively, keep the temperature at 70° C. for constant temperature extraction for 1 h, and then filter to obtain the second extract and the second residue. The weight ratio of panax ginseng and forsythia suspensa separately to the extracting solvent is 1:10.

3. Third Extraction Treatment

[0238] Add ethanol solution at the volume percentage concentration of 70% to the second residue, heat, perform the third heating extraction treatment on panax ginseng and forsythia suspensa, respectively, keep the temperature at 70° C. for constant temperature extraction for 1 h, and then filter to obtain the third extract. The weight ratio of panax ginseng and forsythia suspensa to the extracting solvent is 1:10.

[0239] In the exemplary embodiment of the present invention, in addition to the ethanol solution at the volume percentage concentration of 70% for the extracting solvent used in the second and third extraction processes of the alcohol extract of panax ginseng and forsythia suspensa, other ethanol solutions at a volume percentage concentration of ≥ 50% are applicable to the present invention, preferably ethanol solution at a concentration of 50-75% and absolute alcohol. The extraction temperature of 60-80° C. is applicable to the present invention: The heating extraction time of 0.5-2 h is applicable to the present invention.

[0240] In the exemplary embodiment of the present invention, there are 3 heating extractions in the preparation process of alcohol extract of panax ginseng and forsythia suspensa, and extraction for 2-3 times is applicable to the present invention.

4. Concentration and Drying Treatment

[0241] Mix the first, second and third extracts obtained by 3 filtrations according to the weight ratio of 2: 98, then place it in a rotary evaporator for evaporation and concentration at the temperature of 80° C. to obtain an extractum at the relative density of 1.1, and then place it in an oven for drying to a constant weight at 85° C. to obtain an alcohol extract of the panax ginseng and forsythia suspensa composition.

[0242] The alcohol extract of the panax ginseng and forsythia suspensa composition is a light brownish-yellow powder, has a special odor, and is soluble in ethanol.

[0243] In the process of concentration of the alcohol extract of the panax ginseng and forsythia suspensa composition in the present invention, the concentration temperature of 65-85° C. and the relative density of the concentrated extractum of 1.05-1.15 are applicable to the present invention. The temperature of 70-90° C. in the drying process is applicable to the present invention. The content of total ginsenoside in the alcohol extract of panax ginseng prepared by the method in the present invention is 2-5%.

Exemplary Embodiment 54 Preparation of Water Extract of Panax Ginseng-Forsythia Suspensa Composition

1. Soaking Treatment

[0244] Put the pulverized panax ginseng (300 g), forsythia suspensa (300 g) and extracting solvent tap water (3000 g) into the multi-function extraction tank of traditional Chinese medicine respectively, mix well, and soak for 30 min; The weight ratio of panax ginseng and forsythia suspensa separately to extracting solvent water is 1:10.

[0245] In the preparation process of the water extract of panax ginseng and forsythia suspensa in the present invention, a soaking time of ≥ 30 min is applicable to the present invention, preferably 30-60 min: in addition to the weight ratio of panax ginseng to extracting solvent water of 1:10, other ratios such as 1: 8-12 are applicable to the present invention.

2. First Extraction Treatment

[0246] After soaking for 30 min, turn on the power supply of the multi-function extraction tank, heat, decoct and extract panax ginseng and forsythia suspensa for the first time, keep the temperature at 95° C. for constant temperature extraction for 1 h, and then filter to obtain the first extract and the first residue:

3. Second Extraction Treatment

[0247] Add tap water to the first residue, heat, decoct and extract panax ginseng and forsythia suspensa for the second time, keep the temperature at 90° C. for constant temperature extraction for 1 h, and then filter to obtain the second extract and the second residue. The weight ratio of panax ginseng to extracting solvent is 1:10.

4. Third Extraction Treatment

[0248] Add tap water to the second residue, heat, decoct and extract panax ginseng and forsythia suspensa for the third time, keep the temperature at 90° C. for constant temperature extraction for 1 h, and then filter to obtain the third extract. The weight ratio of panax ginseng and forsythia suspensa to extracting solvent is 1:10.

[0249] In the exemplary embodiment of the present invention, in addition to the weight ratio of 1:10 of panax ginseng to extracting solvent water, other ratios such as 1: 8-12 are applicable to the present invention in the first, second and third extraction processes of the water extract of panax ginseng and forsythia suspensa. The extraction temperature of 90-100° C. is applicable to the present invention, preferably 90-95° C. The heating extraction time of 0.5 to 2 h is applicable to the present invention, preferably 1 h.

[0250] In the exemplary embodiment of the present invention, there are 3 heating extractions in the preparation process of water extract of panax ginseng and forsythia suspensa, and extraction for 2-3 times is applicable to the present invention.

5. Concentration and Drying Treatment

[0251] Mix the first, second and third extracts obtained by 3 filtrations according to the weight ratio of 2: 98. then place it in a rotary evaporator for evaporation and concentration at the temperature of 80° C. to obtain an extractum at the relative density of 1.1, and then place it in an oven for drying to a constant weight at 85° C. to obtain a water extract of panax ginseng.

[0252] The water extract of the panax ginseng and forsythia suspensa composition is a light brownish-yellow powder, has a special odor, and is well water-soluble.

[0253] In the process of concentration of the water extract of the panax ginseng and forsythia suspensa composition in the present invention, a concentration temperature of 70-95° C. and the relative density of a concentrated extractum of 1.05-1.15 are applicable to the present invention: a drying temperature of 70-95° C. is applicable to the present invention.

Exemplary Embodiment 55: Preparation of Total Ginsenoside

[0254] 1. First extraction treatment: the same as in Exemplary Embodiment 5.

[0255] 2. Second extraction treatment: the same as in Exemplary Embodiment 5.

[0256] 3. Third extraction treatment: the same as in Exemplary Embodiment 5.

4. Concentration Treatment

[0257] Combine the first second and third extracts obtained by 3 filtrations, and place it in a rotary evaporator at 80° C. for concentration to obtain panax ginseng concentrate (120 mL), which is equivalent to 2.5 g crude drug/ml panax ginseng solution.

[0258] With respect to the concentration of panax ginseng concentrate in the process of concentrating treatment in the present invention, other concentrations of per milliliter of concentrate equivalent to 1-3.5 g crude drug (i.e. 1-3.5 g crude drug/ml) are applicable to the present invention in addition to the concentration of per milliliter of concentrate equivalent to 2.5 g crude drug.

5. Macroporous Resin Column Chromatography

[0259] Load the panax ginseng concentrate on a macroporous resin column, and perform the macroporous resin column separation treatment, wherein, the D101 macroporous adsorption resin is selected as the macroporous adsorption resin, and the ratio of the resin volume (300 mL) in the resin column to the weight (dry weight) of panax ginseng is 1: 1 (i.e., if the dry weight of panax ginseng is 1 kg. the volume of macroporous resin is 1L: if the dry weight of crude drug is 1 g, the volume of macroporous resin is 1 mL); after the concentrated supernatant completely flows into the resin column, wash with water of 4 times the column volume, and discard the eluate: Then elute with 50% (volume percentage) ethanol solution of 8 times column volume, collect the eluate, and obtain panax ginseng-macroporous resin eluate;

[0260] In the process of macroporous resin column chromatography for the panax ginseng concentrate in the present invention, the ratio of the weight of panax ginseng in the panax ginseng extract to the volume of the macroporous resin of 1: 0.8-2.5 is applicable to the present invention; In addition to D101, other macroporous adsorption resins X-5, AB-8, NK-2, NKA-2, NK-9, D3520. D 101 and WLD are applicable to the present invention: In the process of water elution, the ratio of the amount of water to the column volume of the macroporous resin column of 2-4: 1 is applicable to the present invention. In addition to 50%, other volume percentage concentrations of ethanol solution of 30%-50% are applicable when the eluate is ethanol solution. The ratio of the amount of ethanol solution to the column volume of the macroporous resin column of 2-4: 1 is applicable to the present invention.

6. Concentration and Drying Treatment

[0261] Concentrate the panax ginseng-macroporous resin eluate under reduced pressure at 80° C. in a rotary evaporator, recover the solvent, and dry the concentrated residue in a drying oven at 85° C. to obtain 2.6 g of total ginsenoside.

[0262] Total ginsenoside is a light brownish-yellow powder with a special odor. The total ginsenoside content was measured using the method specified in Appendix B of GB/T19506-2009, and the total ginsenoside content was 32%.

[0263] In the process of concentration of the ginseng-macroporous resin eluate in the present invention, a concentrating temperature of 65-90° C. is applicable to the present invention: A drying temperature of 75-95° C. is applicable to the present invention. The content of total ginsenoside prepared by the method in the present invention is 20-50%.

Exemplary Embodiment 56: Preparation of Panaxadiol Saponins

[0264] 1. First extraction treatment: the same as in Exemplary Embodiment 53.

[0265] 2. Second extraction treatment: the same as in Exemplary Embodiment 53.

[0266] 3. Third extraction treatment: the same as in Exemplary Embodiment 53.

[0267] 4. Concentration treatment: the same as in Exemplary Embodiment 53.

[0268] 5. First macroporous resin column chromatography

[0269] As in Exemplary Embodiment 53, obtain the first panax ginseng-macroporous resin eluate.

[0270] 6. Second macroporous resin column chromatography

[0271] Put the first panax ginseng-macroporous resin eluate to a rotary evaporator for concentration at 80° C. to obtain the first panax ginseng-macroporous resin concentrate (60 ml), which is equivalent to 5.0 g crude drug/ml panax ginseng solution;

[0272] Load the first panax ginseng-macroporous resin concentrate on a macroporous resin column, and perform the macroporous resin column separation treatment, wherein, the HPD-100 macroporous adsorption resin is selected as the macroporous adsorption resin, and the ratio of the resin volume (300 mL) in the resin column to the weight (dry weight) of panax ginseng is 1: 1 (i.e., if the dry weight of panax ginseng is 1 kg, the volume of macroporous resin is IL; if the dry weight of crude drug is 1 g. the volume of macroporous resin is 1 mL); after the concentrated supematant completely flows into the resin column, wash with ethanol solution at the volume percentage concentration of 60% of 4 times the column volume, and discard the eluate; Then elute with 80% (volume percentage) ethanol solution of 8 times column volume, collect the eluate, and obtain the second panax ginseng-macroporous resin eluate.

[0273] With respect to the concentration of panax ginseng concentrate in the process of concentrating treatment in the present invention, other concentrations of per milliliter of concentrate equivalent to 3.5-6 g crude drug (i.e. 3.5-6 g crude drug/ml) are applicable to the present invention in addition to the concentration of per milliliter of concentrate equivalent to 5.0 g crude drug.

[0274] In the process of macroporous resin colunm chromatography of the first panax ginseng-macroporous resin concentrate in the present invention, the ratio of the weight of panax ginseng in the first panax ginseng-macroporous resin concentrate to the volume of the macroporous resin of 1: 0.8-2.5 is applicable to the present invention: In addition to HPD-100. other macroporous adsorption resins such as HPD-200. D203 and XAD-4 are also applicable to the present invention. In the elution process with 60% ethanol solution, the ratio of the amount of 60% ethanol solution to the column volume of the macroporous resin column of 2-4: 1 is applicable to the present invention: In the elution process with 80% ethanol solution, the ratio of the amount of 80% ethanol solution to the column volume of the macroporous resin column of 2-4: 1 is applicable to the present invention.

7. Concentration and Drying Treatment

[0275] Concentrate the second panax ginseng-macroporous resin eluate under reduced pressure at 80° C. in a rotary evaporator, recover the solvent, and dry the concentrated residue in a drying oven at 85° C. to obtain 0.7 g of total panaxadiol saponins.

[0276] Panaxadiol saponins are light yellow powder with a special odor. It has good water solubility. The content of panaxadiol saponins prepared was determined using the high performance liquid chromatography in Appendix VI of Volume I, Chinese Pharmacopoeia (2010 Edition), and the content of panaxadiol saponins was 59%.

[0277] In the process of concentration of the panax ginseng-macroporous resin eluate, a concentration temperature of 65-95° C. is applicable to the present invention; a drying temperature of 70-95° C. is applicable to the present invention. The content of panaxadiol saponins prepared by the method in the present invention is 30-70%.

Exemplary Embodiment 57: Preparation of Forsythin and Phillygenin Composition

[0278] To 1 kg of dry forsythia suspensa leaves, add 10 kg of 95% (m/m) ethanol, heat, reflux and extract 2 times (2 h each time), filter, concentrate the filtrate to ½ of the original volume under reduced pressure, place at 25° C. for 1 h, and allow it to precipitate; Dissolve in methanol, recrystallize, and allow it to precipitate; recrystallize with methanol by repeating the above method, to obtain the amorphous powder of forsythin/phillygenin composition. The contents of forsythin and phillygenin were 98% and 2%, respectively as determined by HPLC.

Exemplary Embodiment 58: Preparation of Forsythin and Phillygenin Composition

[0279] To 1 kg of dry forsythia suspensa, add 10 kg of methanol, heat, reflux and extract 3 times (4 h each time), filter, concentrate the filtrate to ⅒ of the original volume under reduced pressure, place it at 20° C. for 48 h, and allow it to precipitate; Dissolve in ethanol, recrystallize, and allow it to precipitate; recrystallize with ethanol by repeating the above method, to obtain the amorphous powder of forsythin/phillygenin composition. The contents of forsythin and phillygenin were 95% and 4%, respectively.

Exemplary Embodiment 59: Preparation of Forsythin and Phillygenin Composition

[0280] To 1 kg of dry forsythia suspensa leaves, add 10 kg of 70% (m/m) methanol, heat, reflux and extract 3 times (3 h each time), filter, concentrate the filtrate to ⅓ of the original volume under reduced pressure, place at room temperature for 2 h. and allow it to precipitate; Dissolve in 90% methanol, recrystallize, and allow it to precipitate: recrystallize with methanol by repeating the above method, to obtain the amorphous powder of forsythin/phillygenin composition. The contents of forsythin and phillygenin were 88% and 2%, respectively.

Exemplary Embodiment 60: Preparation of Forsythin and Phillygenin Composition

[0281] To 1 kg of dry forsythia suspensa, add 10 kg of absolute ethanol, heat, reflux and extract 2 times (4 h each time), filter, concentrate the filtrate to ¼ of the original volume under reduced pressure, place it at room temperature for 24 h, and allow it to precipitate. Dissolve in acetone, recrystallize, and allow it to precipitate: recrystallize with acetone by repeating the above method to obtain the amorphous powder of forsythin/phillygenin composition. The contents of forsythin and phillygenin were 90% and 6%, respectively.

Exemplary Embodiment 61 Preparation of Forsythin and Phillygenin Composition

[0282] To 1 kg of dry forsythia suspensa leaves, add 10 kg of acetone, heat, reflux and extract 3 times (3 h each time), filter, concentrate the filtrate to ⅕ of the original volume under reduced pressure, place it at room temperature for 10 h, and allow it to precipitate: Dissolve in 70% ethanol, recrystallize, and allow it to precipitate: recrystallize with 70% ethanol by repeating the above method to obtain the amorphous powder of forsythin/phillygenin composition. The contents of forsythin and phillygenin were 80% and 5%, respectively.

Exemplary Embodiment 62-66: Preparation of Composition of Total Ginsenoside- Forsythin and Phillygenin

[0283] Mix the total ginsenoside prepared in Exemplary Embodiment 55 with the forsythin and phillygenin composition prepared in Exemplary Embodiments 57-61 in a weight ratio of 2:98. respectively, to obtain the composition of the total ginsenoside and forsythin.

Exemplary Embodiments 67-71: Preparation of Panaxadiol Saponins- Forsythin and Phillygenin Composition

[0284] Mix the panaxadiol saponins prepared in Exemplary Embodiment 56 with the forsythin and phillygenin composition prepared in Exemplary Embodiments 57-61 in a weight ratio of 2: 98. respectively, to obtain the composition of Total Ginsenoside and forsythin.

Exemplary Embodiment 72: Preparation of Composition of Total Ginsenoside- Forsythin

[0285] Mix the total ginsenoside prepared in Exemplary Embodiment 55 with forsythin in a weight ratio of 2: 98 to obtain the total ginsenoside-forsythin composition.

Exemplary Embodiment 73: Preparation of Panaxadiol Saponins- Forsythin Composition

[0286] Mix the panaxadiol saponins prepared in Exemplary Embodiment 55 with forsythin in a weight ratio of 2: 98 to obtain the panaxadiol saponins - forsythin composition.

Exemplary Embodiment 74: Preparation of 20(R)-Ginsenoside Rg3- Forsythin Composition

[0287] Mix 20(R)-ginsenoside Rg3 and forsythin in a weight ratio of 2: 98 to obtain the composition of 20(R)-ginsenoside Rg3 and forsythin.

Exemplary Embodiments 75-88: Preparation of the Composition of Panax Ginseng-Forsythia Suspensa Composition and Cyclodextrin

[0288] Take the panax ginseng and forsythia suspensa composition prepared in Exemplary Embodiments 53. 54, 62-74, and prepare a composition according to the following method in the said ratio in Table 2: 1) directly add it to the cyclodextrin solution, or 2) directly add it to the cyclodextrin solution and stir thoroughly for 1-24 h, 3) directly add it to the cyclodextrin solution and heat for 10-120 min, 4) directly add it to the cyclodextrin solution and sonicate for 10-120 min, 5) directly grind it with the cyclodextrin powder for 10-120 min. 6) mix the panax ginseng and forsythia suspensa composition with the said cyclodextrin powder well, and sieve; 7) directly add it to the cyclodextrin derivative solution, or 8) directly add it to the cyclodextrin derivative solution and stir thoroughly for 1-24 h, 9) directly add it to the cyclodextrin derivative solution and heat for 10-120 min, 10) directly add it to the cyclodextrin derivative solution and sonicate for 10-120 min, 11) directly grind it with the cyclodextrin derivative powder for 10-120 min. 12) mix it well with the cyclodextrin derivative powder, and sieve.

TABLE-US-00044 Raw Material Ratio and Preparation Method of Exemplary Embodiments 53, 54, 62-74 Exemplary Embodiments No. Panax ginseng component and forsythia suspensa component Grams of composition Grams of cyclodextrin or cyclodextrin derivative Preparation method Exemplary Embodiment 75 100(2:98) 100 1) Directly add Exemplary Embodiment 76 100(2:98) 10000 2) Stir for 1 h Exemplary Embodiment 77 100(2:98) 500 (5) Heat for 10 min Exemplary Embodiment 78 100(2:98) 1000 4) Sonicate for 10 min Exemplary Embodiment 79 100(2:98) 2000 5) Grind for 10 min Exemplary Embodiment 80 100(2:98) 3000 6) Mix well and sieve for 10 min Exemplary Embodiment 81 100(2:98) 4000 1) Directly add Exemplary Embodiment 82 100(2:98) 3500 2) Stir for 12 h Exemplary Embodiment 83 100(2:98) 4500 3) Heat for 120 min Exemplary Embodiment 84 100(2:98) 1500 4) Sonicate for 120 min Exemplary Embodiment 85 100(2:98) 100 (7) Directly add Exemplary Embodiment 86 100(2:98) 10000 (8) Stir for 1 h Exemplary Embodiment 87 100(2:98) 500 (9) Heat for 10 min Exemplary Embodiment 88 100(2:98) 1000 (10) Sonicate for 10 min

[0289] The excipients in Exemplary Embodiments 75-88 are exemplified with selected β-cyclodextrin, and other cyclodextrins and cyclodextrin derivatives are applicable to the present invention. Exemplary Embodiments include 1) β-hydroxypropyl cyclodextrin. 2) hydroxyethyl-β-cyclodextrin, 3) 2,6-dimethyl-β-cyclodextrin, 4) 2,3,6-trimethyl-β-clodextrin, 5) 2,6-diethyl-β-cyclodextrin, 6) 2,3,6-triethyl-β-cyclodextrin, 7) maltosyl-β-cyclodextrin, 8) sulfobutylether β-cyclodextrin, 9) p-toluenesulfonyl chloride (p-TsCl) substituted β-cyclodextrin, 10) 6-position substituted β-CD p-toluenesulfonate (β-cyclodextrin-6-OTs), 11) 2-oxohydroxypropyl-β-cyclodextrin, 12) 2-position monosubstituted p-toluenesulfonate (2-β-cyclodextrin-2-OTs). 13) β-cyclodextrin p-toluenesulfonate (tosy l-β-CD) and 14) PCL-(Tos) 7-β-CD, the star-shaped macromolecule of β-cyclodcxtrin.

Exemplary Embodiment 89: Preparation of Panax Ginseng-forsythia Suspensa Composition Tablets

[0290] Prepare the tablets of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00045 Composition of panax ginseng and forsythia suspensa (weight ratio of 2: 98) 500 g Starch 480 g Talc 1%(10 g) Magnesium stearate 1%(10 g)

[0291] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa prepared in Exemplary Embodiments 53, 54, 62-74 with starch well, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets.

Exemplary Embodiment 90 Preparation of Panax Ginseng-forsythia Suspensa Composition Granules

[0292] Prepare the granules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00046 Composition of panax ginseng and forsythia suspensa (weight ratio of 2: 98) 100 g Microcrystalline cellulose 10000 g

[0293] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa prepared in Exemplary Embodiments 53. 54, 62-74 with microcrystalline cellulose well, granulate, and dispense it into 10000 bags.

Exemplary Embodiment 91: Preparation of Panax Ginseng-forsythia Suspensa Composition Capsules

[0294] Prepare the capsules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00047 Composition of panax ginseng and forsythia suspensa (weight ratio of 2: 98) 250 g Starch 2500 g

[0295] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa prepared in Exemplary Embodiments 53, 54, 62-74 with starch, and fill it into capsules to make 10000 capsules.

Exemplary Embodiments 92-95: Preparation of Panax Ginseng-forsythia Suspensa Composition Capsules

[0296] Mix the components of the panax ginseng and forsythia suspensa composition with starch well according to the weight ratio in Table 3, and fill it into capsules to make 10000 capsules.

TABLE-US-00048 Exemplary Embodiments No. Raw material (Panax ginseng and forsythia suspensa composition, g) Excipient (Starch, g) Weight ratio of raw material to excipient Exemplary Embodiment 92 500(2:98) 500 1:1 Exemplary Embodiment 93 50(10:90) 5000 1:100 Exemplary Embodiment 94 250(25:75) 2500 1:10 Exemplary Embodiment 95 250(50:50) 5000 1:20

[0297] The raw materials in the table may be the compositions of panax ginseng and forsythia suspensa in Exemplary Embodiments 53, 54, 62-74. and complexes of panax ginseng components, forsythia suspensa components and cyclodextrins.

Exemplary Embodiments 96-99: Preparation of Panax Ginseng-forsythia Suspensa Granules

[0298] Mix the components of the panax ginseng and forsythia suspensa composition with microcrystalline cellulose well according to the weight ratio in Table 4. granulate, and dispense it into 10000 bags.

TABLE-US-00049 Exemplary Embodiments No. Raw material (Total ginsenoside and forsythin composition, g) Excipient (Microcrystalline cellulose, g) Weight ratio of raw material to excipient Exemplary Embodiment 96 1000(2:98) 1000 1:1 Exemplary Embodiment 97 250 (10:90) 25000 1:100 Exemplary Embodiment 98 2500(25:75) 25000 1:10 Exemplary Embodiment 99 2500(50:50) 50000 1:20

[0299] The raw materials in the table may be replaced with the compositions of panax ginseng and forsythia suspensa in Exemplary Embodiments 53, 54, 62-74. and complexes of panax ginseng components, forsythia suspensa components and cyclodextrins.

Exemplary Embodiment 100: Preparation of Panax Ginseng-forsythia Suspensa Composition Tablets

[0300] Prepare the tablets of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00050 Composition of panax ginseng and forsythia suspensa (weight ratio of 2: 98) 500 g Starch 380 g Licorice extract 100 g Talc 1%(10 g) Magnesium stearate 1%(10 g)

[0301] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with starch, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa in Exemplary Embodiments 53. 54. 62-74, and complexes of panax ginseng components, forsythia suspensa components and cyclodextrins.

Exemplary Embodiment 101: Preparation of Panax Ginseng-forsythia Suspensa Composition Granules

[0302] Prepare the granules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00051 Composition of panax ginseng and forsythia suspensa (weight ratio of 2: 98) 250 g Licorice extract 250 g Microcrystalline cellulose 24500 g

[0303] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with microcrystalline cellulose, granulate, and dispense it into 10000 bags. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa in Exemplary Embodiments 5, 6. 14-40. and complexes of panax ginseng components, forsythia suspensa components and cyclodextrins.

Exemplary Embodiment 102: Preparation of Panax Ginseng-forsythia Suspensa Composition Capsules

[0304] Prepare the capsules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00052 Composition of panax ginseng and forsythia suspensa (weight ratio of 2:98) 250 g Licorice extract 250 g Extract of fritillariae cirrhosac bulbus 250 g Extract of folium llicis latifoliae 250 g Starch 1000 g

[0305] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with starch, granulate, and fill it into capsules to make 10000 capsules. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa in Exemplary Embodiments 53. 54, 62-74, and complexes of panax ginseng components, forsythia suspensa components and cyclodextrins.

Exemplary Embodiment 103: Preparation of Panax Ginseng-forsythia Suspensa Composition Tablets

[0306] Prepare the tablets of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00053 Composition of panax ginseng and forsythia suspensa (weight ratio of 2:98) 500 g Starch 480 g Extract of anemorrhenae rhizoma 500 g Talc 1%(10 g) Magnesium stearate 1%(10 g)

[0307] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with starch, granulate, add talc and magnesium stearate and mix well, and compress to make 10000 tablets. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa as prepared in Exemplary Embodiments 53. 54, 62-74.

Exemplary Embodiment 104: Preparation of Panax Ginseng-forsythia Suspensa Composition Granules

[0308] Prepare the granules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00054 Composition of panax ginseng and forsythia suspensa (weight ratio of 10:90) 1000 g Extract of scrophulariae radix 500 g Extract loptiatliertiin gracile 500 g Microcrystalline cellulose 10000 g

[0309] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with microcrystalline cellulose, granulate, and dispense it into 10000 bags. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa as prepared in Exemplary Embodiments 53, 54, 62-74.

Exemplary Embodiment 105: Preparation of Panax Ginseng-forsythia Suspensa Composition Capsules

[0310] Prepare the capsules of the composition of panax ginseng and forsythia suspensa according to the following ratio:

TABLE-US-00055 Composition of panax ginseng and forsythia suspensa (weight ratio of 6:94) 2000 g Prunella vulgaris extract 250 g Houttuyniae Herba Extract 500 g Phragmitis rhizoma extract 250 g Starch 1000 g

[0311] According to the above ratio, mix the composition of panax ginseng and forsythia suspensa with the above extract powder well, then mix well with starch, granulate, and fill it into capsules to make 10000 capsules. The composition of panax ginseng and forsythia suspensa in this exemplary embodiment may be replaced with the compositions of panax ginseng and forsythia suspensa as prepared in Exemplary Embodiments 53, 54. 62-74.

Exemplary Embodiment 106: Antiviral Test of Panax Ginseng-Forsythia Suspensa Composition 1 In Vitro Antiviral Test

1.1 Test Materials

Drug

[0312] Forsythin (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. Its purity was 99.5% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area normalization method. The content of forsythin was calibrated and confirmed to be 99.5% using the reference standard of forsythin for assay of pharmaceutical and biological products in China.

[0313] 20(R)-ginsenoside Rg3 (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. Its purity was 99.0% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area normalization method. The content of forsythin was calibrated and confirmed to be 99.1% using the reference standard of ginsenoside Rg3 for the assay of pharmaceutical and biological products in China.

[0314] Panax ginseng and forsythia suspensa composition: Dalian Fusheng Natural Drugs Development Co., Ltd., calibrated it with the reference standard for assay of pharmaceutical and biological products in China, to determine the contents of ginsenoside Rg3 and forsythin.

[0315] Alcohol extract of panax ginseng-forsythia suspensa composition (A: Exemplary Embodiment 53), with the contents of ginsenoside Rg3 and forsythin of 0.15% and 18.6%, respectively: [0316] Water extract of panax ginseng-forsythia suspensa composition (B; Exemplary Embodiment 54), with the contents of ginsenoside Rg3 and forsythin of 0.17% and 17.7%, respectively; [0317] Total ginsenoside-forsythin and phillygenin composition (C; Exemplary Embodiment 62), with the contents of ginsenoside Rg3 and forsythin of 0.04% and 95.6%, respectively; [0318] Panaxadiol saponin-forsythin and phillygenin composition (D; Exemplary Embodiment 67), with the contents of ginsenoside Rg3 and forsythin of 0.12% and 96.6%, respectively; [0319] 20(R)-ginsenoside Rg3-forsythin composition (E; Exemplary Embodiment 74): the content of ginsenoside Rg3 and forsythin is 2.0% and 96.6%, respectively.

[0320] Ribavirin Injection, colorless, transparent liquid, manufactured by Henan Runhong Pharmaceutical Co., Ltd., batch number: 1206261. CFDA Approval No.: H19993553, 100 mg/ml, as the positive control drug in this study.

[0321] Oscetamivir Phosphate. National Institutes for Food and Drug Control. Batch No.: 101090-200901, 100 mg/syringe, as the positive control drug in this study.

[0322] The above drugs were dissolved in corresponding reagents, filtered, sterilized and dispensed, and stored at 4° C. for later use, and are the drugs to be tested in this study.

[0323] Cell strain: Vero (African Green Monkey kidney cells) was preserved by the Basic Medical College of Jilin University.

[0324] Virus strains: 1) Influenza virus, parainfluenza virus and respiratory syncytial virus (RSV) strains were purchased from Institute of Virology, Chinese Academy of Preventive Medicine: 2) Coxsackievirus B.sub.3 (CVB.sub.3) strain: from the United States, preserved by our teaching and research office: 3) Coxsackievirus A16 (CoxA16) and enterovirus 71 (EV71) were gifted by Sendai National Hospital and preserved by our teaching and research office. 4) Adenovirus (AdV): from the Department of Pediatrics, the First Hospital of Norman Bethune Health Science Center, 5) Herpes simplex virus type 1 (HSV-1): purchased from National Institute for the Control of Pharmaceutical and Biological Products, Ministry of Health.

Main Equipment and Reagents

[0325] Biosafety cabinet: BHC-1300 II A/B3, AIRTECH; CO.sub.2 Incubator: MCO-18AIC, SANYO; Inverted microscope: CKX41, OLYMPUS; Electronic analytical balance: AR1140/C, DHAUS; Medium: DMEM. HyClone; Fetal bovine serum: HyClone; Trypsin: Gibco; MTT: Sigma; DMSO: Tianjin Beilian Fine Chemicals Development Co.. Ltd.

1.2 Test Method

Cell Preparation

[0326] Vero cells were subcultured for 1-2 days until they reached confluency. When a clear boundary appeared and strong stereoscopic and refractive index were observed, they were digested with trypsin. When pinpoint-like holes appeared on the cell surface, the digestive fluid was aspirated completely. Then, the cells were split up by several milliliters of culture medium, counted, then diluted to about 5×10.sup.7/L with culture medium (DMEM containing 10% fetal bovine serum), and inoculated into a 96-well culture plate to form a monolayer.

Determination of Drug Toxicity

[0327] Cytotoxicity test: Dilute the drugs at the concentrations shown in Table 1-1 for cytotoxicity determination.

TABLE-US-00056 Reference Table for Drug Dilution (in g/L) Concentration gradient Gradient 1 Gradient 2 Gradient 3 Gradient 4 Gradient 5 Gradient 6 Gradient 7 Gradient 8 Drug Forsythin 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 20(R)-Ginsenoside Rg3 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Composition A 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Composition B 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Composition C 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Composition D 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Composition E 1 0.5 0.25 0.125 0.0625 0.03125 0.015625 0.078125 Ribavirin 5 2.5 1.25 0.625 0.3125 0.15625 0.078125 0.039063 Oseltamivir Phosphate 2 1 0.5 0.25 0.125 0.0625 0.03125 0.015625

[0328] The above drug that was diluted to different concentrations with the cell maintenance medium (DMEM containing 2% fetal bovine serum) was added dropwise to the monolayer of Vero cells, at 0.2 mL per well, in 9 replicate wells for each concentration. Additionally, prepare 9 wells of normal control (blank control group without drug added) and 9 wells of blank control (medium). Then, the cells were cultured in a 5% CO.sub.2 incubator at 37° C. Observation for CPE was performed with an inverted microscope and recorded every day. After 72 h, 20 .Math.L of MTT solution (5 mg/mL) was added to each well, and incubation was continued for 4 h; the medium in each well was aspirated and discarded, and 100 .Math.L of DMSO was added into each well. The plate was shaken for 5 min, the OD value was measured at 492 nm, and the cell survival rate was calculated. Probit regression analysis was performed on the cell viability in the SPSS 18.0 statistical software, and the maximum non-toxic concentration (TC.sub.0) and half toxic concentration (TC.sub.50) of the drug to Vero cells were calculated.

Determination of TCID.SUB.50 of Various Viruses

[0329] The viruses were diluted on a 10-fold basis to different dilutions of 10.sup.-1, 10.sup.-2, 10.sup.-3, 10.sup.-4, 10.sup.-5 and 10.sup.-6 and inoculated in sequence into the monolayer of Vero cells in the 96-well culture plate, at 100 .Math.L per well, in 10 wells for each dilution. Additionally, a normal cell control group was prepared. The plate was incubated in 5% CO.sub.2 at 37° C. for 2 h. and the virus suspension was discarded. Then, 100 .Math.L of cell maintenance medium was added to each well, and the plate was incubated in 5% CO.sub.2 at 37° C. Observation for cytopathic effect was performed with a microscope from Day 3. and the results were interpreted and recorded on Days 7-8. The highest dilution that could cause 50% of the cell wells to show positive cytopathic effect was used as the endpoint, and the virus titer was calculated by Spearman-karber method.

[0330] Formula:

[00001]${\text{LogTCID}}_{50}=\text{XM}{+}^{\frac{1}{2}}{\text{d-d}}^{\frac{{\displaystyle .Math.Pi}}{100}}$

(TCID.sub.50: 50% tissue cell infective dose; XM: logarithm of the dilution of the highest dilution of virus concentration;

[0331] d: logarithm of dilution factor (fold); Σpi: sum of percent cytopathy at each dilution.)

Effect of Drugs on Viral Cytopathic Effect

[0332] The culture plate covered with cell monolayers was taken, and the medium was aspirated and discarded. The cells were inoculated at the virus challenge amount corresponding to 100TCID50, and cell adhesion was allowed in a 5% CO.sub.2 incubator at 37° C. for 2 h. Drug solutions of specific concentrations (maximum non-toxic concentration) were added, and incubation was performed in 10 replicate wells for each concentration at 200 .Math.L/well. Ribavirin Injection and Oseltamivir Phosphate were set as positive drug control groups. Meanwhile, a blank control group (without virus and drug added) and virus control group (with virus added but without drug added) were wet up. Observations were made on the effect of drugs on viral CPE. After 72 h, the OD value was measured by MTT colorimetric assay at the wavelength of 492 nm, and the effective antiviral rate (ER%) was calculated. In the SPSS 18.0 statistical software, the significant differences were analyzed on effective antiviral rate among drugs by ANOVA.

[0333] ER% = (mean OD value of drug treatment group - mean OD value of virus control group)/(mean OD value of cell control group - mean OD value of virus control group) × 100%

1.3 Test Results

TCID.SUB.50 of Various Viruses

[0334] Parainfluenza virus: [0335] Influenza virus: [0336] CVB.sub.3: [0337] HSV-1: [0338] AdV: [0339] RSV: [0340] CoxA16: [0341] EV71:

Determination of Drug Toxicity

1) Determination of Cytotoxicity of Drugs

[0342] The maximum non-toxic concentration (TC.sub.0), half toxic concentration (TC.sub.50) of each drug on Vero cells and the concentration used for the antiviral test are shown in Table 1-2.

TABLE-US-00057 Drug Cytotoxicity Test Results (in g/L) Drug Forsythin 20(R)-Ginsenoside Rg3 Composition A Composition B Virus Maximum non-toxic concentration 0.0066 0.0048 0.026 0.018 Half toxic concentration 0.55 0.31 0.65 0.71 Drug Composition C Composition D Composition E Ribavirin Oseltamivir Phosphate Virus Maximum non-toxic concentration 0.013 0.011 0.016 0.065 0.28 Half toxic concentration 0.71 0.73 0.75 1.392 0.832

2) Results of Drug Protection Against the Viral Cytopathic Effect

[0343] The effective rates of drugs against viruses and the results of one-way analysis of variance by ANOVA method are shown in Table 1-3 and Table 1-4.

TABLE-US-00058 Effective Antiviral Rates (ER%)-Table 1 Drug Forsythin 20(R)-Ginsenoside Rg3 Composition A Composition B Virus Influenza virus 75.38.sup.∗∗ 74.06.sup.∗∗ 86.82.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔ• 89.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔ• Parainfluenza virus 84.96.sup.∗∗ 82.21.sup.∗∗ 89.64.sup.∗∗∗#.triangle-solid. 89.00.sup.∗∗∗#.triangle-solid. CoxA16 75.08.sup.∗∗ 52.33 88.24.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 83.68.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• RSV 80.40.sup.∗∗ 79.42.sup.∗∗ 86.41.sup.∗∗∗.sup.#.triangle-solid.ΔΔ••• 84.32.sup.∗∗∗#.triangle-solid.ΔΔ••• HSV-I 85.00.sup.∗∗ 83.61.sup.∗∗ 88.67.sup.∗∗∗#.triangle-solid.ΔΔ•• 86.64.sup.∗∗∗#.triangle-solid.ΔΔ•• ADV 75.14.sup.∗∗ 51.07 87.51.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 89.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• EV71 84.85.sup.∗∗ 74.66.sup.∗∗ 88.34.sup.∗∗∗#.triangle-solid..triangle-solid.ΔΔΔ••• 89.00.sup.∗∗∗#.triangle-solid..triangle-solid.ΔΔΔ••• CVB.sub.3 75.27.sup.∗∗ 53.75 81.28.sup.∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 85.67.sup.∗∗##.triangle-solid..triangle-solid.ΔΔΔ•••

TABLE-US-00059 Effective Antiviral Rates (ER%)-Table 2 Drug Composition C Composition D Composition E Ribavirin Oseltamivir Phosphate Virus Influenza virus 90.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔ• 100.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔ• 96.82.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔ• 57.49** 81.76** Parainfluenza virus 90.00.sup.∗∗∗#.triangle-solid. 100.00.sup.∗∗∗#.triangle-solid. 99.64.sup.∗∗∗#.triangle-solid. 91.56** 94.52** CoxA16 88.68.sup.∗∗∗.triangle-solid..triangle-solid.ΔΔΔ••• 97.68.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 98.24.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 0.70 2.95 RSV 86.32.sup.∗∗∗#.triangle-solid.ΔΔ••• 95.32.sup.∗∗∗#.triangle-solid.ΔΔ••• 96.41.sup.∗∗∗.triangle-solid.ΔΔ••• 50.08* 37.60 HSV-1 92.64.sup.∗∗∗#.triangle-solid.ΔΔ•• 99.64.sup.∗∗∗#.triangle-solid.ΔΔ•• 98.67.sup.∗∗∗#.triangle-solid.ΔΔ•• 62.92** 66.56** ADV 90.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 100.00.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 97.51.sup.∗∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 0.43 10.31 EV71 90.00.sup.∗∗∗#.triangle-solid..triangle-solid.ΔΔΔ••• 100.00.sup.∗∗∗#.triangle-solid..triangle-solid.ΔΔΔ••• 98.34.sup.∗∗∗#.triangle-solid..triangle-solid.ΔΔΔ••• 4.25 51.86 CVB.sub.3 86.67.sup.∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 92.67.sup.∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 91.28.sup.∗∗##.triangle-solid..triangle-solid.ΔΔΔ••• 13.44 1.64 Note: .sup.∗P < 0.05, .sup.∗∗P < 0.01 compared with the virus control group; .sup.#P < 0.05, .sup.##P < 0.01 for the comparison between panax ginseng-forsythia suspensa composition groups and forsythin; .sup..triangle-solid.P < 0.05, .sup..triangle-solid..triangle-solid.P < 0.01 for the comparison between panax ginseng-forsythia suspensa composition groups and ginsenoside Rg3; .sup.ΔP <0.05, .sup.ΔΔP <0.01, .sup.ΔΔΔP <0.001 for the comparison between panax ginseng-forsythia suspensa composition groups and ribavirin, .sup.•P < 0.05, .sup.••P < 0.01, .sup.•••P < 0.001 for the comparison between panax ginseng-forsythia suspensa composition groups vs. oseltamivir phosphate.

[0344] Tables 1-3 and 1-4 show that the panax ginseng-forsythia suspensa composition groups significantly inhibited 8 kinds of viruses (P < 0.01 or P < 0.001). The effective rates of influenza virus, parainfluenza virus, herpes simplex virus type I (HSV-I), enterovirus 71 and adenovirus (ADV) reached 100%, and the curative effect was significantly better than that of forsythin and ginsenoside Rg3. indicating that the panax ginseng-forsythia suspensa composition groups had a synergistic effect. In addition, the inhibitory effects of panax ginseng-forsythia suspensa composition groups on influenza, coxsackievirus A16 (CoxA16), respiratory syncytial virus (RSV), herpes simplex virus type I (HSV-I), adenovirus (ADV), enterovirus 71 (EV71) and coxsackievirus B.sub.3 (CVB.sub.3) were significantly better than positive drug ribavirin (P < 0.01 or P < 0.001). The inhibitory effects on influenza, coxsackievirus A16 (CoxA16), respiratory syncytial virus (RSV), herpes simplex virus type I (HSV-I), adenovirus (ADV), enterovirus 71 and coxsackievirus B.sub.3 (CVB.sub.3) were significantly better than oseltamivir phosphate (P < 0.05 or P < 0.01, P < 0.001).

2. In Vivo Antiviral Test

2.1 Experimental Materials

Experimental Animals

[0345] Kunming mice were provided by Laboratory Animal Center, Norman Bethune Health Science Center. Jilin University, YDZ No. 10-5219.

Test Instruments and Reagents

[0346] TABLE-US-00060 Name of the Instrument Model Manufacturer Quantitative PCR instrument 7300 ABI PCR instrument ES-60J Shenyang Longteng Electronic Weighing Instrument Co., Ltd. Electronic analytical balance FA1004 Shenyang Longteng Co., Ltd. CO.sub.2 incubator HG303-5 Nanjing Laboratory Instrument Factory Ultra-clean workbench SW-CJ-IF Suzhou Antai Air Technology Co., Ltd. Inverted microscope CKX41 Olympus Instrument -80° C. ultra-low temperature freezer TECON-5082 Australia Water bath oscillator HZS-H Harbin Donglian Co., Ltd. Plate reader TECAN A-5082 Australia Spectrophotometer Model 7550 Japan

2.2 Experimental Method

Determination of Median Lethal Dose of Influenza Virus and Parainfluenza Virus in Mice

[0347] Influenza virus and parainfluenza virus (cell lysate) were serially diluted at 10-fold to the concentrations of 10.sup.-1, 10.sup.-2, 10.sup.-3, 10.sup.-4 and 10.sup.-5. One hundred and twenty Kunming mice were divided into the influenza virus group (n=60) and parainfluenza virus group (n=60), each of which was divided into 6 groups randomly. The mice were lightly anesthetized with ether and intranasally infected with a virus solution of different dilutions at 0.03 mL/animal. Meanwhile, a blank control was set up, where the virus suspension was replaced with normal saline. Observation for mortality and survival was performed daily until 14 days after infection. The death within 24 h of infection was non-specific death, which was not counted. The LD.sub.50 of virus solution was calculated by Karber’s method. Calculation formula:

[00010]${\text{LogLD}}_{50}=\text{XM}{+}^{\frac{1}{2}}{\text{d-d}}^{\frac{{\displaystyle .Math.Pi}}{100}}$

[where: LD.sub.50: median lethal dose; XM: logarithm of the highest dilution of virus concentration: d: logarithm of dilution factor (fold); and Σpi: sum of percent cytopathy at each dilution].

Study on the Effect of Panax Ginseng-Forsythia Suspensa Composition Against the Pulmonitis Induced by Influenza Virus and Parainfluenza Virus Infection

1) Test Animals and Grouping

[0348] Five hundred and forty-four-week-old Kunming mice were selected for two tests.

[0349] Firstly. 270 mice were randomly divided into 27 groups, with 10 mice in each group, and used to determine the lung index and lung index inhibition rate of panax ginseng-forsythia suspensa composition on influenza and parainfluenza virus-infected mice. Ninety mice were selected for each test, and the test was repeated 3 times. Another 270 mice were randomly divided into 27 groups, with 10 mice in each group, which were used to determine the virus hemagglutination titers in lung suspension of panax ginseng-forsythia suspensa composition groups. Ninety mice were selected for each test, and the test was repeated 3 times.

2) Method of Infection

[0350] A mass of absorbent cotton was put in a 200-300 mL beaker, then an appropriate amount of ether was poured (to wet the absorbent cotton), the beaker loaded with absorbent cotton was inverted, and a mouse was placed in the beaker for anesthesia. When the mouse appeared to be extremely excited and then obviously weak, the mouse was placed in a supine position and infected intranasally with 15LD.sub.50 influenza virus and parainfluenza virus at 0.03 ml/nostril. For the blank control group, the virus suspension was replaced with normal saline.

3) Method of Administration and Dosage

[0351] The panax ginseng-forsythia suspensa composition groups, ginsenoside Rg3 group, forsythin group, ribavirin and oseltamivir phosphate control group were adininistrated by gavage on the day before infection, respectively. Panax ginseng-forsythia suspensa composition groups were divided into high, middle and low dose groups, and the dosage was 13.0. 6.5 and 3.25 mg/kg, respectively. The dosage was 13 mg/kg for the forsythin group, 13 mg/kg for the ginsenoside Rg3 group. 58.5 mg/kg for the ribavirin positive drug group and 19.5 mg/kg for the oseltamivir phosphate group, once daily for 5 days. The blank control and virus control groups were administrated by gavage with the same volume of normal saline.

4) Observation Indicators

1) Determination of Lung Index

[0352] On the 5th day after drug administration, the mice were fasted for 8 h, weighed and sacrificed by eyeball removal and exsanguination. The thorax was opened, and the whole lungs were removed, washed twice with normal saline, and the surface was dried with filter paper, and the lungs were weighed by electronic balance. The lung index and lung index inhibition rate were calculated according to the following formulas: [0353] Lung index = (mouse lung weight/mouse body weight) × 100% [0354] Lung index inhibition rate= (mean lung index of infection model group - mean lung index of test group)/mean lung index of infection model group × 100%

2) Determination of Virus Hemagglutination Titer in Lung Suspension

[0355] The lungs of mice in each group were removed on the 5th day after treatment and ground into homogenate with a homogenizer at a low temperature. The homogenate was diluted into 10% lung tissue suspension with normal saline, which was then centrifuged, and the supernatant was taken and diluted on a fold basis. The diluted supernatant was dropped into a titration plate at 0.2 ml/well, and 0.2 ml of 1% chicken red blood cell suspension was added into each well, and the mixture was mixed well and kept at room temperature for 30 min. The hemagglutination titer was observed and recorded. The endpoint was the agglutination of the red blood cells (++), and the titer was expressed by the dilution factor of the suspension.

2.3 Test Results and Analysis

Determination Results of the Median Lethal Dose of Influenza Virus and Parainfluenza Virus in Mice

[0356] Kunming mice in the test group were intranasally infected with 30 .Math.L of influenza virus and parainfluenza virus at different concentrations, respectively. The first 3 groups (virus concentrations of 10.sup.-1, 10.sup.-2 and 10.sup.-3) showed symptoms of varying degrees on day 3 of infection: piloerection, trembling, and reduced food consumption, etc.; On day 5, the mice appeared to wobble while walking. Death was observed in the mice in the highest virus concentration group on day 6, and deaths in the other groups were observed successively on day 7 after infection. At the end of 14-day observation, the number of dead mice in each group was counted, and the results are shown in Tables 1-4 and 1-5 below. The LD.sub.50 was calculated as a dilution of 10.sup.-2.9 for the influenza virus and a dilution of 10.sup.-2.5 for the parainfluenza virus.

TABLE-US-00061 Summary of Median Lethal Dose Test Results of Influenza Virus Influenza virus group Cumulative deaths Cumulative survival Cumulative death rate Group 10-1 9 1 90% Group 10-2 7 3 70% Group 10-3 4 6 40% Group 10-4 3 7 30% Group 10-5 1 9 10% Blank control group 0 10 0%

[0357] The LD.sub.50 of virus was calculated by the Karber method. The LogLD.sub.50 of influenza virus is as follows:

[00011]${\text{LogLD}}_{50}={\text{XM+}}^{\frac{1}{2}}{\text{d-d}}^{\frac{{\displaystyle .Math.Pi}}{100}}=\text{-}1\text{+}0.5\text{-}\left(80\%\text{+}60\%\text{+}40\%\text{+}20\%\text{+}0\%\text{+}0\%\right)=\text{-}2.9$

TABLE-US-00062 Summary of Median Lethal Dose Test Results of Parainfluenza Virus Parainfluenza virus group Cumulative deaths Cumulative survival Cumulative death rate Group 10-1 8 2 80% Group 10-2 6 4 60% Group 10-3 4 6 40% Group 10-4 2 8 20% Group 10-5 0 10 0% Blank control group 0 10 0%

[0358] The LD.sub.50 of virus was calculated by the Karber method. LogLD.sub.50 of parainfluenza virus is as follows:

[00012]${\text{LogLD}}_{50}=\text{XM}{+}^{\frac{1}{2}}{\text{d-d}}^{\frac{{\displaystyle .Math.Pi}}{100}}=\text{-}1\text{+}0.5\text{-}\left(90\%\text{+7}0\%\text{+}40\%\text{+3}0\%\text{+1}0\%\text{+}0\%\right)=\text{-}2.5$

Results of the Effect of Panax Ginseng-forsythia Suspensa Composition Against the Pulmonitis Induced by Influenza Virus and Parainfluenza Virus Infection

1) Determination of Lung Index

[0359] The mean lung index of mice infected with influenza virus and parainfluenza virus showed that compared with the infection model group, the lung index of the blank control group, forsythin group (13.0 mg/kg/d), ginsenoside Rg3 group (16.0 mg/kg/d), panax ginseng-forsythia suspensa composition groups (low dose group 3.25 mg/kg/d, middle dose group 6.5 mg/kg/d, high dose group 13.0 mg/kg/d), ribavirin group and oseltamivir phosphate group decreased significantly (P < 0.05 or P < 0.01). The panax ginseng-forsythia suspensa composition groups had a significant protective effect within the concentration range of 3.25-13.0 mg/kg/d, with significantly reduced lung index and significantly better inhibitory rate on lung tissue lesion index than that of the forsythin group and ginsenoside Rg3 group (P < 0.01 or P < 0.05). See Tables 1-6 and 1-7 for the results.

TABLE-US-00063 Inhibitory Rate of Panax Ginseng-forsythia Suspensa Composition on Lung Index of Mice Infected with Influenza Virus (n=3) Group Drug dose (mg/kg/day) Lung index (x̅ ±S) Lung index Inhibitory rate (%) P-value P-value Blank control group 0 1.274±0.102 — Virus control group 0 1.488±0.084 — Ribavirin 58.5 1.281±0.061 13.90 .sup.∗<0.05 Oseltamivir Phosphate 19.5 1.178±0.066 19.84 .sup.∗∗<0.01 Forsythin 13.0 1.280±0.040 14.00 .sup.∗<0.05 20(R)-Ginsenoside Rg3 13.0 1.302±0.046 12.51 .sup.∗<0.05 Composition A High dose group 13.0 1.078±0.042 28.10 .sup.∗∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.143±0.033 24.00 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Low dose group 3.25 1.191±0.025 19.56 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Composition B High dose group 13.0 1.071±0.046 29.52 .sup.∗∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.133±0.021 24.15 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Low dose group 3.25 1.142±0.027 20.40 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Composition C High dose group 13.0 1.074±0.048 29.16 .sup.∗∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.124±0.031 25.14 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Low dose group 3.25 1.174±0.029 20.56 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Composition D High dose group 13.0 1.049±0.056 29.52 .sup.∗∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.129±0.041 24.15 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Low dose group 3.25 1.184±0.039 20.40 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Composition E High dose group 13.0 1.070±0.056 28.10 .sup.∗∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.131±0.041 24.00 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 Low dose group 3.25 1.197±0.039 19.56 .sup.∗∗<0.01 .sup..triangle-solid.#<0.05 .sup.∗P < 0.05, .sup.∗∗P < 0.01 for the comparison between each test group and the virus control group; .sup.#P < 0.05, .sup.##P < 0.01 for the comparison between panax ginseng-forsythia suspensa composition and forsythin; .sup..triangle-solid.P < 0.05, .sup..triangle-solid..triangle-solid.P < 0.01 for the comparison between panax ginseng-forsythia suspensa composition and ginsenoside Rg3.

TABLE-US-00064 Inhibitory Rate of Panax Ginseng-forsythia Suspensa Composition on Lung Index of Mice Infected with Parainfluenza Virus (n=3) Group Drug dose (mg/kg/day) Lung index (x̅ ±S) Lung index Inhibitory rate (%) P-value P-value Blank control group 0 1.305±0.031 — Virus control group 0 1.591±0.062 — Ribavirin 58.5 1.340±0.065 15.76 .sup.∗<0.05 Oseltamivir Phosphate 19.5 1.243±0.054 21.85 .sup.∗<0.01 Forsythin 13.0 1.335±0.062 16.10 .sup.∗<0.01 20(R)-Ginscnoside Rg3 13.0 1.357±0.050 14.69 .sup.∗<0.01 High dose group 13.0 1.086±0.023 26.74 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Composition A Medium dose group 6.5 1.136±0.047 25.37 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Low dose group 3.25 1.1742±0.036 23.29 .sup.∗<0.01 .sup.#.triangle-solid.0.05 Composition B High dose group 13.0 1.094±0.067 26.87 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.163±0.045 25.43 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Low dose group 3.25 1.181±0.051 22.98 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Composition C High dose group 13.0 1.127±0.033 29.21 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.142±0.034 26.81 .sup.∗<0.01 .sup.#.triangle-solid. .sup.#.triangle-solid..triangle-solid.<0.05 Low dose group 3.25 1.197±0.052 23.62 .sup.∗<0.01 .sup.#.triangle-solid.0.05 Composition D High dose group 13.0 1.068±0.058 32.87 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.143±0.065 28.13 .sup.∗<0.01 .sup.#.triangle-solid. .sup..triangle-solid.<0.05 Low dose group 3.25 1.177±0.044 26.01 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Composition E High dose group 13.0 1.101±0.058 30.79 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Medium dose group 6.5 1.158±0.065 27.22 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Low dose group 3.25 1.188±0.044 25.30 .sup.∗<0.01 .sup.#.triangle-solid.0.05 .sup.∗P < 0.05, .sup.∗∗P < 0.01 for the comparison between each test group and the virus control group; .sup.#P < 0.05, .sup.##P < 0.01 for the companson between panax ginseng-forsythia suspensa composition and forsythin; .sup..triangle-solid.P < 0.05 for the companson between panax ginseng-forsythia suspensa composition and ginsenoside Rg3.

2) Determination of Virus Hemagglutination Titer in Lung Suspension

[0360] The hemagglutination titers (InX) of the virus in lung tissues in the infection model group were 32.40 and 33.11, respectively, after infection with influenza virus and parainfluenza virus in mice. The hemagglutination titers of virus in lung tissues were decreased in the panax ginseng-forsythia suspensa composition groups at different concentrations after 5 days of treatment, and the difference was significant when compared with the infection model group (P < 0.01). The hemagglutination titers of influenza virus and parainfluenza virus with different doses of panax ginseng-forsythia suspensa composition groups were significantly lower than those in the forsythin group and ginsenoside Rg3 group (P < 0.05 to P < 0.001), indicating that the composition had a synergistic effect. The inhibitory rates of the panax ginseng-forsythia suspensa composition groups against virus proliferation were significantly higher than those of the forsythin group and ginsenoside Rg3 group (P < 0.05 to P < 0.001). The inhibitory rates of the high, middle and low dose groups of panax ginseng-forsythia suspensa composition against the hemagglutination titer of the lung suspension of mice infected with influenza virus were significantly higher than those of the Forsythin group and ginsenoside Rg3 group (P < 0.01 to P < 0.001). The results of the above tests are shown in Tables 1-8 and 1-9.

TABLE-US-00065 Effect of Panax Ginseng-forsythia Suspensa Composition on Hemagglutination Titer of Lung Suspension of Mice Infected with Influenza Virus (n=3) Group Drug dose (mg/kg/day) Hemagglutinin titer (InX) Inhibitory rate (%) P-value P-value Blank control group 0 0 Virus control group 0 32.40±1.105 Ribavirin 58.5 21.91±1.050 32.39 .sup.∗<0.01 Oseltamivir Phosphate 19.5 20.50±1.123 36.73 .sup.∗<0.01 Forsythin 13.0 22.06±1.120 31.90 .sup.∗<0.01 20(R)-Ginsenoside Rg3 13.0 22.61±1.059 30.22 .sup.∗<0.01 Composition A High dose group 13.0 18.70±0.428 43.93 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 20.53±0.577 40.02 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Low dose group 3.25 21.03±1.460 32.58 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Composition B High dose group 13.0 18.52±0.572 43.57 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 20.75±0.289 40.31 .sup.∗<0.01 .sup.##.triangle-solid..sup..triangle-solid.<0.01 Low dose group 3.25 21.21±1.398 32.67 .sup.∗<0.01 .sup.#.triangle-solid.<0.05 Composition C High dose group 13.0 18.84±0.364 44.67 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 20.37±0.484 37.25 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Low dose 3.25 21.71±1.217 33.41 .sup.∗<0.01 .sup.#.triangle-solid.<0.05 group Composition D High dose group 13.0 17.86±0.491 43.75 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 18.35±0.397 41.33 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Low dose group 3.25 21.01±1.227 36.59 .sup.∗<0.01 .sup.#.triangle-solid. <0.05 Composition E High dose group 13.0 17.7±0.618 45.36 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 19.21±0.450 40.72 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Low dose group 3.25 20.63±1.439 36.08 .sup.∗<0.01 .sup.#.triangle-solid.<0.05

TABLE-US-00066 Effect of Panax Ginseng-forsythia Suspensa Composition on Hemagglutination Titer of Lung Suspension of Mice Infected with Parainfluenza Virus (n=3) Group Drug dose (mg/kg/day) Hemagglutinin titer (InX) Inhibitory rate (%) P-value P-value Blank control group 0 0 Virus control group 0 33.11±1.210 Ribavirin 58.5 23.22±1.091 29.86 .sup.∗<0.05 Oscltamivir Phosphate 19.5 22.05±1.055 33.40 .sup.∗<0.01 .sup.#.triangle-solid..triangle-solid.<0.05 Forsythin 13.0 23.17±1.059 30.01 .sup.∗<0.01 .sup.#<0.05 20(R)-Ginsenoside Rg3 13.0 23.79±1.072 28.15 .sup.∗<0.01 .sup.#>0.05 Composition A High dose group 13.0 20.1±0.151 43.38 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 21.85±0.550 38.41 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid..triangle-solid.<0.01 Low dose group 3.25 22.81±0.547 35.67 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Composition B High dose group 13.0 19.23±0.480 43.17 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 20.97±0.543 40.80 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid..triangle-solid.<0.01 Low dose group 3.25 22.04±0.457 35.98 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Composition C High dose group 13.0 18.97±0.261 43.98 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 20.80±0.274 39.87 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Low dose group 3.25 21.07±0.613 36.11 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid..triangle-solid.<0.01 Composition D High dose group 13.0 17.38±0.955 47.50 .sup.∗<0.01 .sup.####.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 19.04±0.501 42.49 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid..triangle-solid.<0.01 Low dose group 3.25 20.36±0.824 38.52 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 Composition E High dose group 13.0 17.97±0.955 45.73 .sup.∗<0.01 .sup.###.triangle-solid..triangle-solid..triangle-solid.<0.001 Medium dose group 6.5 19.69±0.501 40.52 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid..triangle-solid.<0.01 Low dose group 3.25 20.81±0.824 37.15 .sup.∗<0.01 .sup.##.triangle-solid..triangle-solid.<0.01 In Tables 1-8 and 1-9, .sup.∗P < 0.05, .sup.∗∗P < 0.01 for the comparison between each test group and the virus control group; .sup.#P < 0.05, .sup.##p < 0.01, .sup.###P < 0.001 for the comparison between panax ginseng-forsythia suspensa composition and forsythin; .sup..triangle-solid.P < 0.05, .sup..triangle-solid..triangle-solid.P< 0.01. .sup..triangle-solid..triangle-solid..triangle-solid.P < 0.001 for the comparison between panax ginseng-forsythia suspensa composition groups and ginsenoside Rg3.

Exemplary Embodiment 107 Antipyretic and Anti-Inflammatory Test of Panax Ginseng and Forsythia Suspensa Composition

1.1 Test Materials

[0361] Test animals Wistar rats, weighing 120-250 g, male and female, certificate No.: YDZ No. 13-1225; Japanese big-ear white rabbits, male, weighing 1.5 ~ 2.0 kg. Certificate No.: YDZ No. 10-5115, all provided by Changchun High-tech Medicine Animal Experimental Research Centre. Animal feed was provided by Laboratory Animal Department of Jilin University.

Test Drugs

[0362] Forsythin (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co.. Ltd. Its purity was 99.5% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area normalization method. The content of forsythin was calibrated and confirmed to be 99.5% using the reference standard of forsythin for the assay of pharmaceutical and biological products in China.

[0363] 20(R)-ginsenoside Rg3 (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. Its purity was 99.0% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area normalization method. The content of forsythin was calibrated and confirmed to be 99.1% using the reference standard of ginsenoside Rg3 for the assay of pharmaceutical and biological products in China.

[0364] Panax ginseng and forsythia suspensa composition: Dalian Fusheng Natural Drugs Development Co.. Ltd., calibrated with the reference standard for assay of pharmaceutical and biological products in China, to determine the contents of ginsenoside Rg3 and forsythin. [0365] Alcohol extract of panax ginseng-forsythia suspensa composition (A; Exemplary Embodiment 53), with the contents of ginsenoside Rg3 and forsythin of 0.15% and 18.6%, respectively: [0366] Water extract of panax ginseng-forsythia suspensa composition (B; Exemplary Embodiment 54), with the contents of ginsenoside Rg3 and forsythin of 0.17% and 17.7%, respectively; [0367] Total ginsenoside-forsythin and phillygenin composition (C; Exemplary Embodiment 62), with the contents of ginsenoside Rg3 and forsythin of 0.04% and 95.6%, respectively; [0368] Panaxadiol saponin-forsythin and phillygenin composition (D; Exemplary Embodiment 67), with the contents of ginsenoside Rg3 and forsythin of 0.12% and 96.6%, respectively; [0369] 20(R)-ginsenoside Rg3-forsythin composition (E; Exemplary Embodiment 74): the content of ginsenoside Rg3 and forsythin is 2.0% and 96.6%, respectively.

1.2 Main Instruments and Reagents

[0370] YLS-7A rat toe swelling measuring instrument: Equipment Station of Shandong Academy of Medical Sciences; 722 visible spectrophotometer: manufactured by Shanghai Spectrum Instruments Co., Ltd.; Portable digital thermometer: model WSC-411P, Shanghai Pudong No.3 Factory; Pilocarpine: Tianjin People’s Pharmaceutical Factory, batch No.: 20130112; Histamine: Shanghai Institute of Biochemistry, batch No.: 20130115; 5-Hydroxytryptamine: Shanghai Institute of Biochemistry, batch No.: 20130623; Evans Blue: Shanghai Chemical Reagent Purchasing and Supply Station, batch No.: 20130217; Chlorpheniramine Tablets: Changchun Economic Development Zone Pharmaceutical Co., Ltd., batch No.: 20130801; Carrageenan: Jilin Institute of Medicine, batch No.: 20130502; Paracetamol Tablets: Liaoyuan Baikang Pharmaceutical Co., Ltd., batch No.: 20130512; Aspirin Tablets: Baicheng Wanda Pharmaceutical Co.. Ltd., batch No.: 20130305: Beer yeast: Beijing Aoboxing Biotech Co., Ltd., batch No.: 2013020; Typhoid and paratyphoid vaccines: Changchun Institute of Biological Products, batch No.: 20130216.

1.3 Statistical Treatment

[0371] Rank sum test. X.sup.2 test and t test were used for statistical analysis.

2.1 Study on the Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Rat Paw (Staining Method)

Materials and Methods

[0372] There are sweat glands on the plantar skin of rats, so the changes in sweat secretion can be observed by the purple reaction between the iodine-starch mixture and the sweat.

[0373] Five hundred Wistar rats, half male and half female, weighing 120-150 g, were taken in the study. They were randomly divided into 50 groups according to body weight and sex, namely the control (0.5% carboxymethylcellulose) group, forsythin group, ginsenoside Rg3 group, low, middle and high dose groups of panax ginseng-forsythia suspensa composition (2.5, 5 and 10 mg/kg. respectively) and positive drug pilocarpine (35 mg/kg) group, with 10 rats in each group, and 10 groups for each test, for totally 5 time periods of testing (1, 5, 10, 15 and 20 min). The rats were placed in a homemade rat-fixing bag to expose both hind limbs. The dirt on the right paw was gently scrubbed with a cotton swab dipped in absolute ethanol. Except for the pilocarpine solution, which was given by subcutaneous injection, other groups were administered by gavage. One hour after administration (30 min after administration in the pilocarpine group), the sweat secreted from the right paw of rats in each group was gently cleaned with a dry cotton swab before the study and during struggling. The right paw was coated with Wotan-Takagaki’s reagent A solution (dissolve 2 g of iodine in 100 mL of absolute ethanol) and then coated with Wotan-Takagaki’s reagent B solution (mix 50 g of soluble starch and 100 mL of Ricinus conununis oil well) after reagent A was fully absorbed. The color and number of dark purple spots (i.e. sweat spots) were observed carefully with a magnifying glass at 1, 5, 10, 15 and 20 min, respectively, after applying solution B. At the end of the test, a rank-sum test was used to analyze the differences among the groups.

Results

[0374] Compared with the control group, the middle and high dose groups (5 and 10 mg/kg) of panax ginseng-forsythia suspensa composition could significantly promote the sweat secretion of rat paw at 10, 15 and 20 min after applying solution B (p < 0.05), and the 2.5 mg/kg group of panax ginseng-forsythia suspensa composition could significantly promote the sweat secretion of rat paw at 15 and 20 min after application of solution B (p < 0.05). The sweating effect was comparable with the positive drug pilocarpine, and the panax ginseng-forsythia suspensa composition could slowly promote the rat paw’s sweat secretion. The high dose group of the panax ginseng-forsythia suspensa composition was significantly superior to forsythin and ginsenoside Rg3 (p < 0.05) in promoting sweat secretion of rat paw following 10, 15 and 20 min after applying solution B. The middle dose group of the panax ginseng-forsythia suspensa composition was significantly superior to forsythin and ginsenoside Rg3 (p < 0.05) in promoting sweat secretion of rat paw following 10 and 15 after application of solution B. The low dose group of the panax ginseng-forsythia suspensa composition was significantly superior to forsythin and ginsenoside Rg3 (p < 0.05) in promoting sweat secretion of rat paw following 15 min after application of B solution. The above test results show that the effect of panax ginseng-forsythia suspensa composition groups was significantly superior to that of forsythin and ginsenoside Rg3 in promoting sweat secretion of rat paw. Please see 2-1. 2-2, 2-3. 2-4, and 2-5 for details.

TABLE-US-00067 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Paw of Normal Rats (Staining Method) Group Animal number (animals) Number of animals with sweat spots of each grade at 1 min after application of solution B (animals) P-value - + ++ +++ +++ Control group 10 2 2 3 2 1 Forsythin 10 0 2 2 2 4 >0.05 Ginsenoside Rg3 10 0 1 3 3 3 >0.05 Pilocarpine 35.0 mg/kg 10 0 1 3 1 5 >0.05 Composition A 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition B 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition C 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition D 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition E 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05

TABLE-US-00068 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Paw of Normal Rats (Staining Method) 2. Animal number 4. Number of animals with sweat spots of each grade at 5 min after application of solution B (animals) 1. Group 3. (animals) - + ++ +++ +++ 5. P-value Control group 10 0 4 1 4 1 Forsythin 10 0 1 2 2 5 >0.05 Ginsenoside Rg3 Pilocarpine 10 0 1 3 2 4 >0.05 35.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ Composition A 2.5 mg/kg 10 0 2 2 1 5 >0.05 5.0 mg/kg 10 0 1 2 2 5 >0.05 10.0 mg/kg 10 0 1 3 2 5 >0.05 Composition B 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition C 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition D 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05 Composition E 2.5 mg/kg 10 0 3 2 0 5 >0.05 5.0 mg/kg 10 0 3 1 1 5 >0.05 10.0 mg/kg 10 0 3 1 1 5 >0.05

TABLE-US-00069 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Paw of Normal Rats (Staining Method) Group Animal number (animals) Number of animals with sweat spots of each grade at 10 min after application of solution B (animals) P-value - + ++ +++ +++ Control group 10 0 3 2 4 1 Forsythin 10 0 1 1 3 5 >0.05 Ginsenoside Rg3 Pilocarpine 10 0 1 3 1 5 >0.05 35.0 mg/kg 10 0 0 2 1 6 <0.05.sup.∗ Composition A 2.5 mg/kg 10 0 0 2 3 5 >0.05 5.0 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 10.0 mg/kg 10 0 1 1 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition B 2.5 mg/kg 10 0 0 2 3 5 >0.05 5.0 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 10.0 mg/kg 10 0 1 1 1 6 <0.05.sup.∗ Composition C 2.5 mg/kg 10 0 0 2 3 5 >0.05 5.0 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 10.0 mg/kg 10 0 1 1 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition D 2.5 mg/kg 10 0 0 2 3 5 >0.05 5.0 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 10.0 mg/kg 10 0 1 1 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition E 2.5 mg/kg 10 0 0 2 3 5 >0.05 5.0 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 10.0 mg/kg 10 0 1 1 1 6 <0.05.sup.∗.sup.#.triangle-solid.

TABLE-US-00070 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Paw of Normal Rats (Staining Method) Group Animal number (animals) Number of animals with sweat spots of each grade at 15 min after application of solution B (animals) P-value - + ++ +++ +++ Control group 10 0 3 2 4 1 Forsythin 10 0 1 2 4 5 <0.05.sup.∗ Ginsenoside Rg3 Pilocarpine 10 0 1 1 3 5 >0.05 35.0 mg/kg 9 0 0 2 1 6 <0.05.sup.∗ Composition A 2.5 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup..triangle-solid. 5.0 mg/kg 10 0 2 1 1 6 <0.05.sup.∗.sup..triangle-solid. 10.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition B 2.5 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup..triangle-solid. 5.0 mg/kg 10 0 2 1 1 6 <0.05.sup.∗.sup..triangle-solid. 10.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition C 2.5 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup.#.triangle-solid. 5.0 mg/kg 10 0 2 1 1 6 <0.05.sup.∗.sup..triangle-solid. 10.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition D 2.5 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.sup..triangle-solid. 5.0 mg/kg 10 0 2 1 1 6 <0.05.sup.∗.sup..triangle-solid. 10.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗.sup.#.triangle-solid. Composition E 2.5 mg/kg 10 0 3 0 1 6 <0.05.sup.∗.triangle-solid. 5.0 mg/kg 10 0 2 1 1 6 <0.05.sup.∗.triangle-solid. 10.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗#.triangle-solid.

TABLE-US-00071 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Paw of Normal Rats (Staining Method) Group Animal number (animals) Number of animals with sweat spots of each grade at 20 min after application of solution B (animals) P-value - + ++ ++ +++ Control group 10 0 3 2 4 1 Forsythin 10 0 0 4 0 6 <0.05* Ginsenoside Rg3 Pilocarpine 10 0 0 1 4 5 <0.05* 35.0 mg/kg 9 0 0 2 1 6 <0.05* Composition A 2.5 mg/kg 10 0 2 1 1 6 <0.05.sup.∗ 5.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ 10.0 mg/kg 10 0 0 2 2 6 <0.05.sup.∗.sup.#.triangle-solid. Composition B 2.5 mg/kg 10 0 2 1 1 6 <0.05.sup.∗ 5.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ 10.0 mg/kg 10 0 0 2 2 6 <0.05.sup.∗.sup.#.triangle-solid. Composition C 2.5 mg/kg 10 0 2 1 1 6 <0.05.sup.∗ 5.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ 10.0 mg/kg 10 0 0 2 2 6 <0.05.sup.∗.sup.#.triangle-solid. Composition D 2.5 mg/kg 10 0 2 1 1 6 <0.05.sup.∗ 5.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ 10.0 mg/kg 10 0 0 2 2 6 <0.05.sup.∗.sup.#.triangle-solid. Composition E 2.5 mg/kg 10 0 2 1 1 6 <0.05.sup.∗ 5.0 mg/kg 10 0 1 2 1 6 <0.05.sup.∗ 10.0 mg/kg 10 0 0 2 2 6 <0.05.sup.∗♯.triangle-solid. Evaluation standard for sweat spot grade: “-”: there is no sweat spot on the surface of plantar pad of the rat, “+”: sweat spots are occasionally found on the surface of the plantar pad of the rat, and the area of sweat spots was less than 10% of the plantar surface; “++”: sweat spots are scattered on the surface of the plantar pad of the rat, and the area of sweat spots is about 11-40% of the plantar surface; “+++”: sweat spots are distributed on multiple areas of the surface of the plantar pad of the rat, and the area of sweat spots is about 41-70% of the plantar surface; “++++”: sweat spots are distributed evenly on the surface of the plantar pad of the rat and the area of sweat spots is about more than 71% of the plantar surface.

[0375] *P < 0.05 for the comparison between each test group and the virus control group; .sup.#P < 0.05 for the comparison between panax ginseng-forsythia suspensa composition and forsythin; .sup..triangle-solid.P < 0.05 for the comparison between panax ginseng-forsythia suspensa composition and ginsenoside Rg3.

2.2 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Rat Paw (Histomorphology Observation Method)

Materials and Methods

[0376] After sweat glands activation in rats, besides the increase of sweat secretion, the morphology of the epithelial cells of the sweat glands changes accordingly. Under the optical microscope, the increase and enlargement of vacuoles in the epithelial cells of the sweat glands are observed. Under an electron microscope, the enlarged vacuoles appear to be due to swelling and rupture of mitochondria, enlargement of fusion and secretory vesicles in the epithelial cells of sweat glands. The secretory activity of sweat glands could be understood by observing the histomorphology of the epithelial cells of plantar sweat glands of rats.

[0377] Three hundred Wistar rats, half male and half female, weighing 120-160 g. were taken in the study. They were randomly divided into 30 groups according to body weight and sex, namely the control group (0.5% carboxymethylcellulose), ginsenoside Rg3, forsythin, low, middle and high dose groups of panax ginseng-forsythia suspensa composition (2.5, 5, 10 mg/kg) and positive drug pilocarpine group (35 mg/kg), with 10 rats in each group, and 3 tests for each group. Except for the pilocarpine solution given by subcutaneous injection, other groups were administered by gavage. One hour after the control group was given 0.5% carboxymethyl cellulose, 30 min after the positive drug group was given pilocarpine, and 1 h after the administration of forsythin, ginsenoside Rg3 and the panax ginseng-forsythia suspensa composition, the right hind limb was immediately cut off at the level of the ankle joint. Then, the right plantar pad was removed, placed in 10% formaldehyde solution, fixed, dehydrated, embedded, sectioned and stained with HE. Observations were made under an optical microscope for changes in the epithelial cells of the plantar sweat glands of rats in each group, mainly the incidence of vacuoles. The statistical processing was performed by X.sup.2 test, and differences among the groups were compared. The above study was repeated for 3 times.

[00013]$\begin{array}{l}\text{Percentage}\text{of}\text{vacuolation}=\\ \text{number}\text{of}\text{vacuolar}\text{sweat}\text{glands}/\text{number}\text{of}\text{observed}\text{sweat}\text{glands}\times \\ 100\%\end{array}$

Results

[0378] Compared with the control group, the 2.5 mg/kg. 5 mg/kg, 10 mg/kg groups of panax ginseng-forsythia suspensa composition very significantly promoted the sweat secretion of rat paw (p < 0.001). The low, middle and high dose groups (2.5, 5 and 10 mg/kg) had significantly better efficacy than forsythin and ginsenoside Rg3 (p < 0.001 or p < 0.01), indicating that the compositions of panax ginseng and forsythia suspensa have a synergistic effect. The test results are shown in Tables 2-6;

TABLE-US-00072 Effect of the Panax Ginseng and Forsythia Suspensa Composition on Sweat Secretion of Rat Paw (Histomorphology Observation Method, n=3) Group Animal number (animals) Number of observed sweat glands (number) Number of vacuolar sweat glands (number) Vacuolation rate (%) Control group 10 242 14 5.78 Forsythin 10 209 86 22.15.sup.∗∗ Ginsenoside Rg3 10 212 79 20.26.sup.∗∗ Pilocarpine 35.0 mg/kg’ 10 208 57 27.40.sup.∗∗∗ Composition A 2.5 mg/kg 10 221 66 29.86.sup.∗∗∗##.triangle-solid..triangle-solid. 5.0 mg/kg 10 215 73 33.95.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. 10.0 mg/kg 10 207 82 39.61.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. Composition B 2.5 mg/kg 10 221 68 30.76.sup.∗∗∗##.triangle-solid..triangle-solid. 5.0 mg/kg 10 218 71 32.56.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. 10.0 mg/kg 10 209 87 41.62.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. Composition C 2.5 mg/kg 10 222 65 29.27.sup.∗∗∗##.triangle-solid..triangle-solid. 5.0 mg/kg 10 217 72 33.17.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. 10.0 mg/kg 10 208 83 39.03.sup.∗∗∗###.triangle-solid..triangle-solid..triangle-solid. Composition D 2.5 mg/kg 10 221 25 30.31.sup.∗∗∗.sup.##.triangle-solid..triangle-solid. 5.0 mg/kg 10 213 73 34.27.sup.∗∗∗.sup.###.triangle-solid..triangle-solid..triangle-solid. 10.0 mg/kg 10 207 85 41.63.sup.∗∗∗.sup.###.triangle-solid..triangle-solid..triangle-solid. Composition E 2.5 mg/kg 10 225 66 29.30.sup.∗∗∗.sup.##.triangle-solid..triangle-solid. 5.0 mg/kg 10 219 69 31.51.sup.∗∗∗.sup.###.triangle-solid..triangle-solid..triangle-solid. 10.0 mg/kg 10 208 87 41.82.sup.∗∗∗.sup.###.triangle-solid..triangle-solid..triangle-solid. .sup.∗∗p<0.01, .sup.∗∗∗p<0.001 compared with the control group; .sup.##P < 0.01. .sup.###P < 0.001 for the companson between panax ginseng-forsythia suspensa composition and forsythin; .sup..triangle-solid..triangle-solid.P <0.05 0.01, .sup..triangle-solid..triangle-solid..triangle-solid.P <0.05 0.001 for the comparison between panax ginseng-forsythia suspensa composition groups and ginsenoside Rg3.

2.3 Effect of the Panax Ginseng-forsythia Suspensa Composition on the Fever Induced by Beer Yeast in Rats

Materials and Methods

[0379] Male Wistar rats: weighing 180-200 g. Before the test, the normal rectal temperature was measured twice with WSC411P portable digital thermometer (at a certain interval), and the mean value of the two measurements was taken as the normal body temperature of the rats. Then. 300 rats with a body temperature of 36.5 ~ 38° C. were randomly divided into 30 groups: the model group (0.5% carboxymethylcellulose), low, middle and high dose groups of panax ginseng-forsythia suspensa composition (2.5, 5, 10 mg/kg), forsythin group (13 mg/kg), ginsenoside Rg3 group (13 mg/kg) and positive drug paracetamol group (100 mg/kg), with 10 rats in each group, and 3 replicate tests for each group. The rats in each group were subcutaneously injected with 10% fresh beer yeast suspension at 10 mL/kg on the back to induce fever. Six point zero hours after administration of 10% fresh beer yeast suspension, the composition of ginsenoside Rg3 and forsythin and paracetamol were given by gavage, and the model group was given an equal volume of 0.5% carboxymethylcellulose by gavage. Rectal temperature was measured at 1, 2, 3 and 4 h after administration. Observations were made on changes in body temperature, and the differences among the groups were compared by t-test of antipyretic percentage. The above study was repeated 3 times.

[00014]$\begin{array}{l}\text{Antipyretic}\text{percentage}=\\ \text{Body}\text{temperature}\text{at}\text{a}\text{certain}\text{time}\text{after}\text{administration}\text{-}\\ \text{body}\text{temperature}\text{6h}\text{after}\text{fever}\text{induction}\\ \text{Body}\text{temperature}\text{at}\text{6h}\text{after}\text{induction}\\ \times 100\%\end{array}$

Results

[0380] Six hours after subcutaneous injection of 10% fresh beer yeast suspension to rats in each group, the body temperature increased by about 1.5° C., which was significantly different from that before fever induction (p < 0.001), indicating that the rat fever model induced by beer yeast was established successfully. Compared with the model group, a significant temperature-lowering effect on the rat fever induced by beer yeast suspension was noted in the middle and high dose groups of panax ginseng-forsythia suspensa composition at 1, 2, 3 and 4 h after drug administration and in the low dose group of panax ginseng-forsythia suspensa composition at 2, 3 and 4 h after drug administration (p<0.05 to p<0.001). Meanwhile, the different doses of panax ginseng-forsythia suspensa composition were significantly superior to the forsythin group and ginsenoside Rg3 group (p < 0.001 or p < 0.01) in lowering the temperature, indicating that they had a significant synergistic effect. The results of the above study are shown in Tables 2-7.

2.4 Effect of the Panax Ginseng-Forsythia Suspensa Composition on Carrageenan-Induced Paw Swelling in Rats

Materials and Methods

[0381] Two hundred and twenty male Wistar rats weighing 120-150 g were randomly divided into 22 groups, namely the control group (0.5% sodium carboxymethylcellulose), low, middle and high dose groups of panax ginseng-forsythia suspensa composition (2.5. 5, 10 mg/kg), forsythin group, 20(R)-ginsenoside Rg3 group and positive drug aspirin group (200 mg/kg), with 10 rats in each group. Each group was administered by sublingual intravenous injection. Before the study, the normal volume of the right hind limb was measured by the capillary magnification method. To avoid errors, the measurement position should be fixed and performed by one person before and after administration. The average of the two measurements was taken as the normal volume of the right hind limb of the rats prior to administration. Immediately after administration, 0.1 ml of 1% carrageenan was subcutaneously injected into the right hind paw of rats to induce inflammation. The right hind plantar volume was measured at 15, 30, 60, 120, 180, 240, 300 and 360 min after inflammation induction. The inter-group t-test was performed by the difference percentage (swelling rate) of the plantar volume before and after the inflammation induction, and the differences among the groups were compared.

[00015]$\begin{array}{l}\text{Swelling}\text{rate}\left(\%\right)=\\ \text{Right}\text{hind}\text{plantar}\text{volume}\text{after}\text{inflammation}\text{induction}\text{-}\\ \text{right}\text{hind}\text{plantar}\text{volume}\text{before}\text{administration}\\ \text{Right}\text{plantar}\text{volume}\text{before}\text{administration}\\ \times 100\%\end{array}$

Results

[0382] Compared with the blank control group, the rat paw swelling induced by carrageenan was significantly inhibited (p < 0.05 or p < 0.01) in the high dose group (10 mg/kg) of panax ginseng-forsythia suspensa composition within 15 min to 360 min after administration, the middle dose group (5 mg/kg) and the low dose group (2.5 mg/kg) within 30 min to 360 min after administration, and the efficacy was significantly better than that of the forsythin 10 mg/kg group and 20(R)-ginsenoside Rg3 10 mg/kg group (p < 0.05 or p < 0.01). The efficacy of the above-mentioned dose groups of the composition was very significantly better than that of the 20(R)-ginsenoside Rg3 group at 60 min and 240 min after administration (p < 0.01). The above study results showed a significant synergistic effect when forsythin in the panax ginseng-forsythia suspensa composition was combined with 20(R)-ginsenoside Rg3. See Table 2-8 for details.

TABLE-US-00073 Effect of the Panax Ginseng-forsythia Suspensa Composition on the Fever Induced by Beer Yeast in Rats [00016]$\left({}^{\overline{\text{χ}}}\pm \text{s},\text{n}=3\right)$ Group Normal 6 h after fever induction Body temperature (°C) Time post-dose (h) 1 2 3 4 Model control group (%) 37.72±0.90 39.30±0.54 39.44±0.58 39.42±0.47 38.88±0.46 38.57±0.49 4.22±2.11.sup.### 0.37±1.66 0.31±1.77 -1.07±1.54 -1.86±1.20 Paracetamol 100 mg/kg(%) 37.55±0.70 39.48±0.62 38.66±0.59 38.19±0.59 37.98±0.19 37.84±0.32 5.14±1.42.sup.### -2.07±0.54.sup.∗∗ -3.27±0.77.sup.∗∗∗ -3.80±1.43.sup.∗∗∗ -4.15±1.59.sup.∗∗∗ Forsythin 10 mg/kg (%) 37.58±0.59 39.53±0.63 39.13±0.52 38.77±0.42 38.63±0.40 38.46±0.31 5.18±1.52.sup.### -0.60±0.39 -0.92±0.93.sup.∗∗ -1.29±1.18.sup.∗ -1.70±1.23.sup.∗ 20(R)-Ginsenoside Rg3 10 mg/kg (%) 37.50±0.59 39.44±0.47 39.31±0.48 39.04±0.45 38.91±0.40 38.70±0.31 5.17±1.37.sup.##.sup.# -0.3±0.41 -0.69±0.93.sup.∗ -1.03±1.18.sup.∗ -1.55±1.23.sup.∗ Composition A 2.5 mg/kg(%) 37.46±0.34 39.57±0.55 39.22±0.49 38.94±0.42 38.47±0.41 38.27±0.49 5.14±1.38.sup.### -0.62±1.04.sup..triangle-solid. -1.88±1.04.sup..triangle-solid. -2.38±1.17.sup.∗∗Δ.triangle-solid. -3.17±1.74.sup.∗∗∗Δ.triangle-solid. 5.0 mg/kg(%) 37.52±0.36 39.48±0.52 38.79±0.52 38.37±0.25 38.28±0.33 37.97±0.41 5.36±1.18.sup.### -1.77±0.48.sup.∗Δ.triangle-solid..triangle-solid. -2.15±0.45.sup.∗∗Δ.triangle-solid. -2.52±0.39.sup.∗∗Δ.triangle-solid. -3.62±0.54.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. 10.0 mg/kg(%) 37.38±0.67 39.52±0.49 38.69±0.38 38.34±0.47 38.21±0.73 37.87±0.28 5.21±1.44.sup.### -2.03±0.46.sup.∗∗ΔΔ.triangle-solid..triangle-solid. -2.46±0.65.sup.∗∗Δ.triangle-solid..triangle-solid. -2.94±1.08.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. -3.78±1.12.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. Composition B 2.5 mg/kg(%) 37.57±0.41 39.49±0.36 31.21±0.48 38.94±0.42 38.47±0.54 38.31±0.48 5.45±1.28.sup.### -0.67±1.26.sup..triangle-solid. -1.23±1.54.sup..triangle-solid. -1.95±1.08.sup.∗∗Δ.triangle-solid. -2.87±1.73.sup.∗∗∗Δ.triangle-solid. 5.0 mg/kg(%) 37.52±0.43 39.56±0.38 38.79±0.51 38.43±0.42 38.28±0.37 37.93±0.47 5.29±1.16.sup.### -1.76±0.47.sup.∗Δ.triangle-solid..triangle-solid. -2.18±0.54.sup.∗∗Δ.triangle-solid. -2.84±0.31.sup.∗∗Δ.triangle-solid. -3.74±0.38.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. 10.0 mg/kg(%) 37.67±0.29 39.48±0.53 38.72±0.37 38.27±0.65 38.18±0.50 37.87±0.42 5.09±1.27.sup.### -1.63±0.45.sup.∗∗ΔΔ.triangle-solid..triangle-solid. -2.37±0.62.sup.∗∗Δ.triangle-solid..triangle-solid. -3.15±1.10.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. -4.10±1.32.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. Composition C 2.5 mg/kg(%) 37.64±0.42 39.60±0.63 39.26±0.47 38.97±0.41 38.59±0.35 38.30±0.50 5.21±1.40.sup.### -0.87±1.12.sup..triangle-solid. -1.59±1.18.sup..triangle-solid. -2.54±1.32.sup.∗∗Δ.triangle-solid. -3.29±1.81.sup.∗∗∗Δ.triangle-solid. 5.0 mg/kg(%) 37.47±0.63 39.47±0.43 38.76±0.56 38.87±0.52 38.35±0.40 37.95±0.38 5.33±1.27.sup.### -1.80±0.50.sup.∗Δ.triangle-solid..triangle-solid. -2.27±0.50.sup.∗∗Δ.triangle-solid. -2.84±0.31.sup.∗∗Δ.triangle-solid. -3.85±0.44.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. 10.0 mg/kg(%) 37.55±0.76 39.48±0.67 38.71±0.49 38.39±0.48 38.24±0.37 37.90±0.31 5.13±1.50.sup.### -1.95±0.58.sup.∗∗ΔΔ.triangle-solid..triangle-solid. -2.76±0.85.sup.∗∗Δ.triangle-solid..triangle-solid. -3.15±1.10.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. -4.00±1.12.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. Composition D 2.5 mg/kg(%) 37.33±0.51 39.23±0.63 38.86±0.47 38.20±0.44 37.85±0.52 38.52±0.59 5.10±1.45.sup.### -0.95±1.02.sup..triangle-solid. -1.70±1.11.sup..triangle-solid. -2.61±1.65.sup.∗∗Δ.triangle-solid. -3.45±1.88.sup.∗∗Δ.triangle-solid. 5.0 mg/kg(%) 37.41±0.55 39.37±0.41 38.61±0.52 38.46±0.55 38.22±0.32 37.84±0.41 5.25±1.25.sup.### -1.92±0.42.sup.∗Δ.triangle-solid..triangle-solid. -2.31±0.57.sup.∗∗Δ.triangle-solid. -2.93±0.36.sup.∗∗Δ.triangle-solid. -3.93±0.50.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. 10.0 mg/kg(%) 37.53±0.59 39.54±0.62 38.69±0.57 37.72±0.40 37.27±0.42 37.08±0.35 5.36±1.52.sup.### -2.16±0.51.sup.∗∗ΔΔ.triangle-solid..triangle-solid. -2.85±0.93.sup.∗∗Δ.triangle-solid..triangle-solid. -3.66±1.18.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. -4.16±1.23.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. Composition E 2.5 mg/kg(%) 37.58±0.47 39.68±0.36 39.25±0.33 18.46±0.57 38.11±0.47 38.30±0.50 5.17±1.28.sup.### -0.43±1.09.sup..triangle-solid. -1.07±1.05.sup..triangle-solid. -2.06±1.28.sup.∗∗.sup.Δ.triangle-solid. -3.29±1.81.sup.∗∗∗.sup.Δ.triangle-solid. 5.0 mg/kg(%) 37.63±0.67 39.56±0.48 38.67±0.46 38.13±0.46 37.85±0.46 37.95±0.38 5.21±1.36.sup.### -1.69±0.62.sup.∗.sup.Δ.triangle-solid..triangle-solid. -2.15±0.48.sup.∗∗.sup.Δ.triangle-solid. -2.32±0.44.sup.∗∗.sup.Δ.triangle-solid. -3.85±0.44.sup.∗∗∗.sup.ΔΔ.triangle-solid..triangle-solid. 10.0 mg/kg(%) 37.55±0.52 39.62±0.53 38.59±0.55 37.95±0.48 37.82±0.33 37.63±0.29 5.25±1.44.sup.### -1.95±0.58.sup.∗∗ΔΔ.triangle-solid..triangle-solid. -2.68±0.58.sup.∗∗Δ.triangle-solid..triangle-solid. -3.36±1.04.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. -4.13±1.24.sup.∗∗∗ΔΔ.triangle-solid..triangle-solid. .sup.∗p <0.05, .sup.∗∗p< 0.01, .sup.∗∗∗p < 0.001 compared with the model control group; .sup.###p < 0.001 compared with normal (before fever induction). Δp < 0.05; ΔΔp<0.01; ΔΔΔp<0.001 for the comparison of antipyretic percentage between panax ginseng-forsythia suspensa composition and foresythin; .sup..triangle-solid.p < 0.05; .sup..triangle-solid..triangle-solid.p < 0.01; .sup..triangle-solid..triangle-solid..triangle-solid.p < 0.001 for the comparison of antipyretic percentage between panax ginseng-forsythia suspensa composition and ginsenoside Rg3.

TABLE-US-00074 Inhibitory Effect of the Panax Ginsenig-forsythia Suspensa Composition on Carrageenan-induced Paw Swelling in Rats [00017]$\left({}^{\overline{\text{χ}}}\pm \text{s},\text{n}=10\right)$ Group Swelling rate (%) 15 min 30 min 60 min 120 min 180 min 240 min 300 min 360 min Control group 18.7±12.6 28.9±14.6 33.8±10.5 46.5±18.2 46.7±15.3 48.8±21.9 49.1±14.6 47.4±15.5 Aspinn 100 mg/kg 8.90±6.70.sup.∗ 15.6±12.2.sup.∗ 20.3±10.9.sup.∗ 21.6±12.1.sup.∗∗ 26.1±21.1.sup.∗∗ 26.1±16.3.sup.∗∗ 34.9±14.6.sup.∗ 31.9±12.2.sup.∗ Forsythin 10.0 mg/kg 12.9±8.41 19.9±9.55 21.8±11.8* 28.9±14.0.sup.∗ 30.8±15.4.sup.∗ 29.9±11.1.sup.∗ 33.5±10.0.sup.∗ 32.2±11.9.sup.∗ 20(R)-Ginsenoside Rg3 10.0 mg/kg 14.9±6.13 21.9±9.72 27.1±13.5 30.7±10.2.sup.∗ 33.8±12.1.sup.∗ 31.0±11.3.sup.∗ 35.8±13.5.sup.∗ 36.4±10.2 2.5 mg/kg 14.8±8.74 18.9±12.1 23.5±13.3.sup.∗.triangle-solid..triangle-solid. 28.9±14.2.sup.∗∗Δ.triangle-solid. 31.8±14.8.sup.∗∗Δ.triangle-solid. 33.1±18.3.sup.∗∗Δ.triangle-solid. 34.9±19.6.sup.∗∗Δ.triangle-solid. 37.8±18.3.sup.∗.triangle-solid..triangle-solid. Composition A 5.0 mg/kg 12.7±8.66 15.1±7.15*.sup.Δ.triangle-solid. 17.5±8.43.sup.∗.triangle-solid..triangle-solid. 19.2±10.4.sup.∗∗Δ.triangle-solid. 22.6±12.9.sup.∗∗Δ.triangle-solid. 24.6±13.7.sup.∗∗Δ.triangle-solid. 27.3±11.7.sup.∗∗Δ.triangle-solid. 30.3±16.8.sup.∗∗Δ.triangle-solid..triangle-solid. 10.0 mg/kg 10.4±8.12.sup.∗Δ.triangle-solid. 12.4±9.52.sup.∗Δ.triangle-solid. 14.3±11.9.sup.∗∗Δ.triangle-solid..triangle-solid. 16.6±10.7.sup.∗∗Δ.triangle-solid. 19.6±12.7.sup.∗∗Δ.triangle-solid. 21.5±11.6.sup.∗∗Δ.triangle-solid. 24.2±12.7.sup.∗∗Δ.triangle-solid. 28.9±15.1.sup.∗∗Δ.triangle-solid..triangle-solid. 2.5 mg/kg 14.5±9.11 17.4±16.8 22.1±17.3.sup.∗.triangle-solid..triangle-solid. 27.8±13.5.sup.∗∗Δ.triangle-solid. 30.9±16.5.sup.∗∗Δ.triangle-solid. 33.4±17.3.sup.∗∗Δ.triangle-solid. 33.6±15.8.sup.∗∗Δ.triangle-solid. 36.9±21.0.sup.∗.triangle-solid..triangle-solid. Composition B 5.0 mg/kg 13.1±8.97 16.7±7.62.sup.∗Δ.triangle-solid. 18.5±10.35.sup.∗.triangle-solid..triangle-solid. 21.6±13.7.sup.∗∗Δ.triangle-solid. 23.9±14.7.sup.∗∗Δ.triangle-solid. 26.8±12.8.sup.∗∗Δ.triangle-solid. 28.9±10.6.sup.∗∗Δ.triangle-solid. 32.7±13.9.sup.∗∗Δ.triangle-solid..triangle-solid. 10.0 mg/kg 10.3±7.65.sup.∗Δ.triangle-solid. 12.4±9.88.sup.∗Δ.triangle-solid. 15.1±13.8.sup.∗∗Δ.triangle-solid..triangle-solid. 17.8±12.9.sup.∗∗Δ.triangle-solid. 20.1±13.7.sup.∗∗Δ.triangle-solid. 25.5±13.6.sup.∗∗Δ.triangle-solid. 26.2±15.1.sup.∗∗Δ.triangle-solid. 29.4±14.6.sup.∗∗Δ.triangle-solid..triangle-solid. 2.5 mg/kg 13.5±8.52 20.9±16.0 21.1±14.1.sup.∗.triangle-solid..triangle-solid. 23.1±16.5.sup.∗∗Δ.triangle-solid. 27.4±15.6.sup.∗∗Δ.triangle-solid. 27.7±23.7.sup.∗∗Δ.triangle-solid. 30.4±16.9.sup.∗∗Δ.triangle-solid. 33.1±22.8.sup.∗.triangle-solid..triangle-solid. Composition C 5.0 mg/kg 12.4±8.71 16.0±6.15.sup.∗Δ.triangle-solid. 19.5±9.55.sup.∗.sup..triangle-solid..triangle-solid. 20.2±11.6.sup.∗∗Δ.triangle-solid. 23.2±15.2.sup.∗∗Δ.triangle-solid. 25.6±11.6.sup.∗∗Δ.triangle-solid. 28.89±12.7.sup.∗∗Δ.triangle-solid. 31.6±15.5.sup.∗∗Δ.triangle-solid..triangle-solid. 10.0 mg/kg 9.9±7.80.sup.∗Δ.triangle-solid. 13.9±9.61.sup.∗Δ.triangle-solid. 16.9±12.2.sup.∗∗Δ.triangle-solid. 18.2±14.8.sup.∗∗Δ.triangle-solid. 20.5±15.9.sup.∗∗Δ.triangle-solid. 23.8±12.0.sup.∗∗Δ.triangle-solid. 26.1±11.3.sup.∗∗Δ.triangle-solid. 29.8±11.9.sup.∗∗Δ.triangle-solid..triangle-solid. Composition D 2.5 mg/kg 13.6±8.40 24.2±16.3 20.1±14.8.sup.∗.triangle-solid..triangle-solid. 22.0±16.0.sup.∗∗Δ.triangle-solid. 26.8±16.1.sup.∗∗Δ.triangle-solid. 26.7±23.3.sup.∗∗Δ.triangle-solid. 29.5±16.1.sup.∗∗Δ.triangle-solid. 32.4±23.4.sup.∗.triangle-solid..triangle-solid. 5.0 mg/kg 11.4±8.90 15.2+6.20.sup.*Δ.triangle-solid. 18.0±9.00.sup.∗.triangle-solid..triangle-solid. 19.7±12.1.sup.∗∗Δ.triangle-solid. 22.0±14.9.sup.∗∗Δ.triangle-solid. 24.2±12.0.sup.∗∗Δ.triangle-solid. 27.8±13.1.sup.∗∗Δ.triangle-solid. 30.0±16.0.sup.∗∗Δ.triangle-solid..triangle-solid. 10.0 mg/kg 9.0±7.90.sup.∗Δ.triangle-solid. 13.2±9.50.sup.∗Δ.triangle-solid. 15.8±11.6.sup.∗∗Δ.triangle-solid..triangle-solid. 17.9±14.3.sup.∗∗Δ.triangle-solid. 19.7±15.2.sup.∗∗Δ.triangle-solid. 22.5±11.8.sup.∗∗Δ.triangle-solid. 25.4±10.6.sup.∗∗Δ.triangle-solid. 28.7±11.4.sup.∗∗Δ.triangle-solid..triangle-solid. 2.5 mg/kg 12.7±8.26 19.5±14.06 20.9±13.7.sup.∗.triangle-solid..triangle-solid. 22.9±14.2.sup.∗∗Δ.triangle-solid. 24.6±12.4.sup.∗∗Δ.triangle-solid. 26.8±16.7.sup.∗∗Δ.triangle-solid. 29.5±13.7.sup.∗∗Δ.triangle-solid. 32.1±18.7.sup.∗.triangle-solid..triangle-solid. Composition E 5.0 mg/kg 11.3±7.92 14.3±7.33.sup.∗Δ.triangle-solid. 16.5±8.62.sup.∗.triangle-solid..triangle-solid. 18.7±13.9.sup.∗∗Δ.triangle-solid. 22.7±14.2.sup.∗∗Δ.triangle-solid. 24.3±14.1.sup.∗∗Δ.triangle-solid. 26.1±19.3.sup.∗∗Δ.triangle-solid. 29.6±16.1.sup.∗∗Δ.triangle-solid..triangle-solid. 10.0 mg/kg 9.6±7.45.sup.∗Δ.triangle-solid. 12.6±9.54.sup.∗Δ.triangle-solid. 15.8±11.3.sup.∗∗Δ.triangle-solid..triangle-solid. 17.4±12.6.sup.∗∗Δ.triangle-solid. 19.1±12.8.sup.∗∗Δ.triangle-solid. 21.8±16.3.sup.∗∗Δ.triangle-solid. 24.7±16.3.sup.∗∗Δ.triangle-solid. 27.8±15.4.sup.∗∗Δ.triangle-solid..triangle-solid. .sup.∗p <0.05, .sup.∗∗p < 0.01 compared with the control group; Δp < 0.05, ΔΔp<0.01 for the comparison of antipyretic percentage between panax ginseng-forsythia suspense composition and forsythin; .triangle-solid.p< 0.05, .triangle-solid..triangle-solid.p < 0.01 for the comparison of antipyretic percentage between panax ginseng-forsythia suspensa composition and ginsenoside Rg3.

Exemplary Embodiment 108 Exploration on the Effect of the Panax Ginseng-Forsythia Suspensa Composition on Enhancing Immune Function in Mice

1. Experimental Materials

1.1 Drugs and Reagents

[0383] Forsythin (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co.. Ltd. Its purity was 99.5% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area nonnalization method. The content of forsythin was calibrated and confirmed to be 99.5% using the reference standard of forsythin for the assay of pharmaceutical and biological products in China.

[0384] 20(R)-ginsenoside Rg3 (content > 98%), white powder, is manufactured by Dalian Fusheng Natural Drugs Development Co., Ltd. Its purity was 99.0% as determined by HPLC with two detectors: UV detector and evaporative light scattering detector using the area normalization method. The content of forsythin was calibrated and confirmed to be 99.1% using the reference standard of ginsenoside Rg3 for the assay of pharmaceutical and biological products in China.

[0385] Panax ginseng and forsythia suspensa composition: Dalian Fusheng Natural Drugs Development Co., Ltd., calibrated with the reference standard for assay of pharmaceutical and biological products in China, to determine the contents of ginsenoside Rg3 and forsythin. [0386] Alcohol extract of panax ginseng-forsythia suspensa composition (A; Exemplary Embodiment 53), with the contents of ginsenoside Rg3 and forsythin of 0.15% and 18.6%, respectively; [0387] Water extract of panax ginseng-forsythia suspensa composition (B; Exemplary Embodiment 54), with the contents of ginsenoside Rg3 and forsythin of 0.17% and 17.7%, respectively; [0388] Total ginsenoside-forsythin and phillygenin composition (C: Exemplary Embodiment 62), with the contents of ginsenoside Rg3 and forsythin of 0.04% and 95.6%, respectively; [0389] Panaxadiol saponin-forsythin and phillygenin composition (D; Exemplary Embodiment 67), with the contents of ginsenoside Rg3 and forsythin of 0.12% and 96.6%, respectively; [0390] 20(R)-ginsenoside Rg3-forsythin composition (E: Exemplary Embodiment 74): the content of ginsenoside Rg3 and forsythin is 2.0% and 96.6%, respectively. [0391] Positive control drug: Pidotimod Oral Solution (Suzhou Pharmaceutical Factory of Jiangsu Wuzhong Pharmaceutical Group Corporation, strength: 10 mL: 400 mg, batch number: 2014091211);

1.2 Experimental Animals

[0392] Kunming mice, aged 6-8 weeks, weighing 18-22 g. were purchased from Experimental Animal Center of Dalian Medical University, with quality certificate No.: SCXK (13) 2013-0003.

2. Experimental Method

2.1 Grouping and Administration

[0393] One hundred and fifty-two healthy male mice were subjected to acclimation for 4 days before they were randomly divided into 19 groups: the negative control group, positive control group, forsythin group, 20(R)-ginsenoside Rg3 group, high, middle and low dose groups of panax ginseng-forsythia suspensa composition. The positive control groups were given pidotimod (50 mg/kg), forsythin (144 mg/kg), and 20(R)-ginsenoside Rg3 (144 mg/kg); the panax ginseng-forsythia suspensa composition groups were given low dose (36 mg/kg), middle dose (72 mg/kg) and high dose (144 mg/kg). The drugs were given by gavage, once daily for 30 days, and the negative control group was given the same volume of water.

2.2 Experiments and Results

2.2.1 ConA-induced Mouse Splenic Lymphocyte Transformation Test

[0394] At 1 h after the last dose, the spleens of animals of each group were aseptically excised to prepare splenocyte suspension. After diluting the splenocyte suspension to a concentration of 3×10.sup.6 cells/mL, the splenocyte suspension was divided into 2 aliquots.Each of the aliquot was transferred into two 24-well culture plates at a volume of 1 mL/well followed by addition of 75 .Math.L of ConA solution (a) was added to one well, and the other well was used as control (b). The wells were cultured at 37° C. for 72 h. Then, thiazolyl blue (MTT) was added 4 h before the end of the culture. After incubation, acidic isopropanol was added, and the absorbance (ABS) of each solution was measured at 570 nm after it was mixed well. The proliferativity was calculated using the formula: proliferativity = ABSa-ABSb. Each dose group of the test sample was compared with the negative control group. The experimental results are shown in Table 1.

2.2.2 Determination of NK Cell Viability

[0395] One hour after the last dose, the spleens of animals of each group were aseptically excised to prepare splenocyte suspension. After the red blood cells were lysed with sterile water for injection, the cell suspension was diluted with 1% glacial acetic acid. Then, the concentration of splenocyte suspension was adjusted to 2×10.sup.7 cells/mL. and the splenocyte suspension was added into a 96-well plate for culturing. For each animal, the wells were divided into reaction wells (splenocyte suspension and YAC-1 cell suspension 100 .Math.L each, effector-to-target ratio 50:1), natural release wells (YAC-1 cell suspension and culture medium 100 .Math.L each) and maximum release wells (YAC-1 cell suspension and 1% NP40 100 .Math.L each). A triplicate set of tubes was prepared for each of the above items. The 96-well plate was incubated at 37° C. in a 5% CO.sub.2 incubator for 4 h. After adding LDH matrix solution and 1 moL/L HCl, the solutions in each parallel well were combined, and the absorbance (ABS) was measured at 490 nm. The NK cell viability was calculated: NK cell viability (%) = (ABS .sub.reaction .sub.well -ABS .sub.natural .sub.release .sub.well)/(ABS .sub.maximum .sub.release .sub.well -ABS .sub.natural .sub.release .sub.well). The experimental results are shown in Table 1.

TABLE-US-00075 Effect of the Panax Ginseng-forsythia Suspensa Composition on NK Cell Viability and ConA-induced Lymphocyte Proliferation Ability (x±s) Group Dose administered (mg/kg) NK cell viability (%) Proliferation ability (×10.sup.-2 ) Negative control group -- 34.14±5.68 9.04±3.81 Positive control group 50 42.57±7.31 27.97±11.45 Forsythin 144 41.86±7.24 28.66±12.63 20(R)-Ginsenoside Rg3 144 37.34±7.21 23.9±11.70 Composition A Low dose group 36 38.77±9.15 20.65±8.03 Medium dose group 72 41.52±10.53 25.61±10.12 High dose group 144 45.77±8.76 33.89±16.87 Composition B Low dose group 36 38.38±6.84 19.94±10.60 Medium dose group 72 40.31±11.29 26.08±11.74 High dose group 144 44.82±7.97 31.84±13.61 Composition C Low dose group 36 39.64±7.15 20.89±7.84 Medium dose group 72 43.21±10.18 28.03±12.02 High dose group 144 45.62±11.74 35.54±13.97 Composition D Low dose group 36 40.13±9.25 21.96±7.38 Medium dose group 72 44.33±9.51 30.55±10.91 High dose group 144 47.43±8.27 37.85±12.93 Composition E Low dose group 36 40.75±7.51 20.93±9.11 Medium dose group 72 44.79±11.64 29.49±12.52 High dose group 144 47.92±8.94 37.13±17.03

3. Test Results

[0396] Following the stimulation of T lymphocytes by ConA, the blast cells will have proliferative responses. The mitochondrial hydrolase in the viable cells, especially the proliferating cells, decomposes MTT into blue-purple crystals. Increased optical density value, if any, indicates that the cell proliferation ability is enhanced. As shown in Table 1, the optical density differences of the high, middle and low dose groups of the panax ginseng-forsythia suspensa composition were higher than those of the negative control group, indicating that this sample had the effect of promoting the proliferation of splenocytes.

[0397] After the cells are killed by the NK cells, the LDH in the cytoplasm of viable cells will be released out of the cells, dehydrogenate lithium lactate and reduce NAD to NADH, and NADH is further reduced to iodonitro-tetrazolium chloride (INT) through hydrogen transmitter phenazine methosulfate (PMS). After receiving H+, the INT is reduced to a purple formazan compound, and the optical density value is determined using a microplate reader. As shown in Table 1, the NK cell viability of the high, middle and low dose groups of the panax ginseng-forsythia suspensa composition was significantly higher than that of the negative control group, indicating that the sample could increase the viability of the NK cells.

[0398] The above study results showed that different doses of the panax ginseng-forsythia suspensa composition could promote the proliferation of spleen cells and enhance the activity of NK cells. Moreover, the composition of panax ginseng and forsythia suspensa had a synergistic effect, and could significantly promote the proliferation of spleen cells, enhance the activity of NK cells and improve immune function.