CN112618661A - Antiviral traditional Chinese medicine composition and preparation method and application thereof - Google Patents
Antiviral traditional Chinese medicine composition and preparation method and application thereof Download PDFInfo
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Abstract
An antiviral traditional Chinese medicine composition specifically comprises the following raw materials: the traditional Chinese medicine composition can be widely applied to influenza virus, parainfluenza virus, hepatitis B virus and hepatitis C virus, has obvious curative effect and low toxicity, and provides a new effective weapon for treating influenza caused by influenza parainfluenza virus and hepatitis caused by hepatitis B or hepatitis C virus.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to an antiviral traditional Chinese medicine composition as well as a preparation method and application thereof.
Background
Radix tetrastigme is a Vitaceae plant, also named as Anoectochilus roxburghii Hance, radix tetrastigmae, Shimaoji, XIAOTENG, SHIHOU, TUJING pill, and radix Trifolii Pratense. The main producing areas are in the southwest of China, Anhui, Zhejiang, Jiangxi, Fujian, Hubei, Hunan, Guangdong and Guangxi.
The radix tetrastigme is used as a medicine by tubers and whole herbs, has the effects of improving immunity, resisting virus and enhancing liver function, and is commonly used for treating febrile convulsion, rheumatic aching pain, irregular menstruation, cough, asthma, purulent phlegm, sore throat, scrofula, carbuncle, furuncle and other symptoms clinically. Has obvious curative effect on serious infectious diseases such as pneumonia, tuberculous lymphadenitis, hepatitis, gastritis, cervicitis, septicemia and the like. Has wide and good therapeutic effect on various malignant tumors, hypertension, heart disease, diabetes, neurasthenia and other diseases. But the related research of applying the radix tetrastigme to the antiviral drugs is less at present.
Therefore, the technical personnel in the field need to solve the problem of providing the traditional Chinese medicine composition containing the radix tetrastigme, which has antiviral effect and good curative effect.
Disclosure of Invention
In view of the above, the invention provides an antiviral traditional Chinese medicine composition and a preparation method thereof, the antiviral traditional Chinese medicine composition is prepared by combining radix tetrastigme, dendrobium officinale and liquorice, the dendrobium officinale adopted by the invention has the effect of improving the immunity of an organism and can be used together with glycyrrhizic acid, glycyrrhetinic acid and radix tetrastigme in the liquorice to have a synergistic antiviral effect, the traditional Chinese medicine composition can be widely applied to influenza virus, parainfluenza virus, hepatitis B virus and hepatitis C virus, and has the advantages of obvious curative effect, low toxicity and obvious progress.
In order to achieve the purpose, the invention adopts the following technical scheme:
an antiviral traditional Chinese medicine composition specifically comprises the following raw materials: 2-3 parts of radix tetrastigme, 1 part of dendrobium officinale and 1 part of liquorice.
Preferably, the following raw materials are specifically included: the traditional Chinese medicine composition comprises, by weight, 4 parts of radix tetrastigme, 2 parts of dendrobium officinale and 2 parts of liquorice.
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing radix tetrastigme ultrafine powder: weighing 2-3 parts of raw material radix tetrastigme, and crushing into superfine powder for later use;
(4) sterilizing the dendrobium officinale extract, the liquorice extract and the radix tetrastigme ultrafine powder respectively, and mixing uniformly to obtain the antiviral traditional Chinese medicine composition.
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing an ethanol extract of radix tetrastigme: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40% -70% ethanol, boiling, refluxing and extracting for 2-4h for 3 times, then filtering, combining filtrates, and concentrating to obtain radix tetrastigme ethanol extract for later use;
(4) and uniformly mixing the dendrobium officinale extract, the liquorice extract and the radix tetrastigme alcohol extract, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing a mixed solution I of radix tetrastigme alcohol extraction and water extraction: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40% -70% ethanol, boiling, reflux extracting for 2-4h for 3 times, filtering, reserving filter residue, combining filtrates, and concentrating to obtain radix tetrastigme ethanol extract; adding water into the residue, boiling, reflux-extracting for 2-4 hr for 3 times, filtering, mixing filtrates, and concentrating to obtain radix Apioris Fortunei water extractive solution; mixing the radix tetrastigme alcohol extract and the radix tetrastigme water extract to obtain a radix tetrastigme alcohol extraction and water extraction mixed solution I;
(4) and uniformly mixing the dendrobium officinale extracting solution, the liquorice extracting solution and the radix tetrastigme alcohol extraction and water extraction mixed solution I, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing a mixed solution II of radix tetrastigme alcohol extraction and water extraction: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40% -70% ethanol, boiling, refluxing and extracting for 2-4h for 3 times, then filtering, reserving filter residues for later use, and combining the filtrate to obtain radix tetrastigme ethanol extract; adding water into the residue, boiling, reflux-extracting for 2-4 hr for 3 times, filtering, and mixing filtrates to obtain radix Apioris Fortunei water extractive solution; mixing the radix tetrastigme alcohol extract and the radix tetrastigme water extract, refrigerating overnight, centrifuging, and filtering to obtain a radix tetrastigme alcohol extraction and water extraction mixed solution II;
(4) and uniformly mixing the dendrobium officinale extracting solution, the liquorice extracting solution and the radix tetrastigme alcohol extraction and water extraction mixed solution II, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
Preferably, the volume of the water in the step (1) is 15-30 times of the mass of the dendrobium officinale; the volume of the water in the step (2) is 10-15 times of the mass of the liquorice.
Preferably, the concentration in the step (1) is carried out until every 5g of the dendrobium officinale extract contains 1g of crude drugs.
Preferably, the concentration in step (1) is performed until the licorice extract solution contains 1g of crude drug per 1g of concentrated solution.
Preferably, the concentration of the mixed solution of ethanol extraction and water extraction of radix tetrastigme I in the step (3) is 0.5-0.75g of crude drug per ml of mixed solution.
Preferably, the concentration of the mixed solution II of ethanol extraction and water extraction of radix tetrastigme in the step (3) is 0.5-0.75g of crude drug per ml of mixed solution.
Preferably, the mesh number of the ultrafine powder in the step (3) is 200 meshes.
Preferably, the 40-70% ethanol and the radix tetrastigme in the step (3) are in a volume-to-mass ratio.
Preferably, the sterilizing conditions of the dendrobium officinale extracting solution and the liquorice extracting solution in the step (4) are as follows: sterilizing at 121 deg.C for 20-30 min; the radix tetrastigme ultrafine powder is sterilized by irradiating with 25-50KGY cobalt 60-gamma rays for 18-22 min.
Preferably, the sterilization conditions in the step (4) are sterilization at 121 ℃ for 20-30 min.
The traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method is applied to antiviral drugs, and the viruses are influenza viruses, parainfluenza viruses, hepatitis B viruses and hepatitis C viruses.
According to the technical scheme, compared with the prior art, the invention has the following beneficial effects: the invention provides an antiviral traditional Chinese medicine composition and a preparation method thereof, the antiviral traditional Chinese medicine composition is prepared by combining radix tetrastigme, dendrobium officinale and liquorice, the traditional Chinese medicine composition can be widely applied to influenza viruses, parainfluenza viruses, hepatitis B viruses and hepatitis C viruses, has obvious curative effect and low toxicity compared with various chemical medicines in the prior art, and provides a new effective weapon for treating influenza caused by influenza parainfluenza viruses and hepatitis caused by hepatitis B or hepatitis C viruses.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a line graph showing the specific inhibitory effect of ribavirin and 4 samples on H1N1 PR8 infection and toxicity results on MDCK cells in the pharmacodynamic experiment of the present invention;
FIG. 2 is a microscopic view showing the results of the specific inhibitory effect of ribavirin and 4 samples on H1N1 PR8 infection and toxicity to MDCK cells in the pharmacodynamic experiment of the present invention;
FIG. 3 is a line graph showing the specific inhibitory effect of ribavirin and 4 samples on HPIV3 parainfluenza virus and the toxicity results on MK2 cells in the pharmacodynamic experiment of the present invention;
FIG. 4 is a microscopic image showing the specific inhibitory effect of ribavirin and 4 samples on HPIV3 parainfluenza virus and the toxicity results on MK2 cells in the pharmacodynamic experiment of the present invention;
FIG. 5 is a line graph showing the specific inhibitory effect of 2' CMC and 4 samples on HCV virus and the toxicity results on Huh7.5.1 cells in the pharmacodynamic experiment of the present invention;
FIG. 6 is a microscopic view showing the specific inhibitory effect of entecavir and 4 samples on HBV virus and the toxicity result on HepAD38 cell in the pharmacodynamic experiment of the present invention;
FIG. 7 is a line graph showing the results of toxicity of entecavir and 4 samples against HBeAg and HBsAg of HBV on cells in the pharmacodynamic experiment of the present invention;
FIG. 8 is a line graph showing the results of the inhibition of HBVDNA and the toxicity to cells by entecavir and 4 samples in the pharmacodynamic test of the present invention;
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 15 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 4 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, adding water with the volume 10 times of the mass of the liquorice, heating, boiling, refluxing and extracting for 3 times, extracting for 1 hour each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and concentrating under reduced pressure to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing radix tetrastigme ultrafine powder: weighing 300g of radix tetrastigme, and crushing into 200-mesh ultrafine powder for later use;
(4) pouring herba Dendrobii extract and Glycyrrhrizae radix extract into a clean glass bottle, adding water with the same mass as radix Apioris Fortunei superfine powder, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling; placing radix Apioris Fortunei micropowder into a clean glass bottle, sealing with a gland, and sterilizing by 25-50KGY cobalt 60-gamma ray irradiation for 20 min; mixing the sterilized herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei micropowder to obtain antiviral Chinese medicinal composition.
Example 2
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 30 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 8 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, heating, boiling and carrying out reflux extraction on the water with the volume 15 times of the mass of the liquorice for 3 times, extracting for 3 hours each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and carrying out reduced pressure concentration to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing radix tetrastigme ultrafine powder: weighing 200g of radix tetrastigme, and crushing into 200-mesh ultrafine powder for later use;
(4) pouring herba Dendrobii extract and Glycyrrhrizae radix extract into a clean glass bottle, adding water with the same mass as radix Apioris Fortunei superfine powder, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling; placing radix Apioris Fortunei micropowder into a clean glass bottle, sealing with a gland, and sterilizing by 25-50KGY cobalt 60-gamma ray irradiation for 20 min; mixing the sterilized herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei micropowder to obtain antiviral Chinese medicinal composition.
Example 3
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 15 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 4 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, adding water with the volume 10 times of the mass of the liquorice, heating, boiling, refluxing and extracting for 3 times, extracting for 1 hour each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and concentrating under reduced pressure to finally obtain the liquorice extracting solution for later use;
(3) preparing an ethanol extract of radix tetrastigme: weighing 300g of raw material radix tetrastigme, adding 40% ethanol with the volume 5 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 2h for 3 times, then centrifugally filtering the extracting solution (4000rpm, 20min), merging the filtrate, and concentrating to obtain the radix tetrastigme ethanol extract for later use; the radix tetrastigme alcohol extract concentration is 0.5 g crude drug per ml;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extract into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Example 4
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 30 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 8 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, heating, boiling and carrying out reflux extraction on the water with the volume 15 times of the mass of the liquorice for 3 times, extracting for 3 hours each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and carrying out reduced pressure concentration to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing an ethanol extract of radix tetrastigme: weighing 200g of raw material radix tetrastigme, adding 70% ethanol with the volume 15 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 4h for 3 times, then centrifugally filtering the extracting solution (4000rpm, 20min), merging the filtrate, and concentrating to obtain the radix tetrastigme ethanol extract for later use; the radix tetrastigme alcohol extract concentration is 0.5 g crude drug per ml;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extract into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Example 5
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 15 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 4 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, adding water with the volume 10 times of the mass of the liquorice, heating, boiling, refluxing and extracting for 3 times, extracting for 1 hour each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and concentrating under reduced pressure to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing an ethanol extract of radix tetrastigme: weighing 300g of raw material radix tetrastigme, adding 40% ethanol with the volume 5 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 2h for 3 times, then carrying out centrifugal filtration (4000rpm, 20min) on the extracting solution, filtering residues for later use, combining the filtrates, and concentrating to obtain an ethanol extract of the radix tetrastigme; adding 8-16 times of water into the residue, boiling, reflux extracting for 3 times (2 hr each time), centrifuging the extractive solution (4000rpm for 20min), mixing filtrates, and concentrating under reduced pressure to obtain radix Apioris Fortunei water extractive solution; mixing radix Apioris Fortunei alcohol extract and radix Apioris Fortunei water extract to obtain radix Apioris Fortunei alcohol extraction and water extraction mixed solution I; the concentration of the mixed solution I of radix tetrastigme alcohol extraction and water extraction is 0.5 g crude drug per milliliter;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extraction + water extraction mixed solution I into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Example 6
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 30 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 8 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, heating, boiling and carrying out reflux extraction on the water with the volume 15 times of the mass of the liquorice for 3 times, extracting for 3 hours each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and carrying out reduced pressure concentration to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing an ethanol extract of radix tetrastigme: weighing 200g of radix tetrastigme as raw material, adding 70% ethanol with the volume 15 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 4h for 3 times, then carrying out centrifugal filtration (4000rpm for 20min) on the extracting solution, filtering residues for later use, combining the filtrates, and concentrating to obtain the radix tetrastigme ethanol extract; adding 8-16 times of water into the residue, boiling, reflux extracting for 3 times (4 hr each time), centrifuging the extractive solution, filtering (4000rpm for 20min), mixing filtrates, and concentrating under reduced pressure to obtain radix Apioris Fortunei water extractive solution; mixing radix Apioris Fortunei alcohol extract and radix Apioris Fortunei water extract to obtain radix Apioris Fortunei alcohol extraction and water extraction mixed solution I; the concentration of the mixed solution I of radix tetrastigme alcohol extraction and water extraction is 0.5 g crude drug per milliliter;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extraction + water extraction mixed solution I into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Example 7
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the volume 15 times of the mass of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 4 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, adding water with the volume 10 times of the mass of the liquorice, heating, boiling, refluxing and extracting for 3 times, extracting for 1 hour each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and concentrating under reduced pressure to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing an ethanol extract of radix tetrastigme: weighing 300g of radix tetrastigme as raw material, adding 40% ethanol with the volume 5 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 2h for 3 times, then carrying out centrifugal filtration (4000rpm, 20min) on the extract, keeping filter residues for later use, and combining the filtrates to obtain the radix tetrastigme ethanol extract; adding 8-16 times of water into the residue, boiling, reflux extracting for 3 times (2 hr each time), centrifuging the extractive solution (4000rpm, 20min), and mixing filtrates to obtain radix Apioris Fortunei water extractive solution; mixing radix Apioris Fortunei alcohol extract and radix Apioris Fortunei water extract, refrigerating at-4 deg.C overnight, centrifuging, and filtering to obtain radix Apioris Fortunei alcohol extract and water extract; the concentration of the mixed solution II of the radix tetrastigme alcohol extraction and the water extraction is that each milliliter contains 0.5 g of crude drug;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extraction and water extraction mixture into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Example 8
The preparation method of the antiviral traditional Chinese medicine composition specifically comprises the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 100g of dendrobium officinale, crushing into small segments of 1-2cm, flattening, adding water with the mass of 30 times of the volume of the dendrobium officinale, heating, boiling, refluxing and extracting for 3 times, extracting for 8 hours each time, filtering the extracting solution through a 100-mesh and 200-mesh filter screen, combining the filtrates, and concentrating under reduced pressure to finally obtain the dendrobium officinale extracting solution for later use; the herba Dendrobii extract contains crude drug 1g per 5 g;
(2) preparing a liquorice extracting solution: weighing 100g of liquorice, crushing into 20-60 meshes of coarse powder, heating, boiling and carrying out reflux extraction on the water with the volume 15 times of the mass of the liquorice for 3 times, extracting for 3 hours each time, filtering the extracting solution by a 100-mesh and 200-mesh filter screen, merging the filtrate, and carrying out reduced pressure concentration to finally obtain the liquorice extracting solution for later use; the Glycyrrhrizae radix extract contains 1g crude drug per 1 g;
(3) preparing an ethanol extract of radix tetrastigme: weighing 200g of radix tetrastigme as raw material, adding 70% ethanol with the volume 15 times of the mass of the radix tetrastigme, boiling, refluxing and extracting for 4h for 3 times, then carrying out centrifugal filtration (4000rpm, 20min) on the extract, keeping filter residues for later use, and combining the filtrates to obtain the radix tetrastigme ethanol extract; adding 8-16 times of water into the residue, boiling, reflux extracting for 3 times (4 hr each time), centrifuging the extractive solution (4000rpm, 20min), and mixing filtrates to obtain radix Apioris Fortunei water extractive solution; mixing radix Apioris Fortunei alcohol extract and radix Apioris Fortunei water extract, refrigerating at-4 deg.C overnight, centrifuging, and filtering to obtain radix Apioris Fortunei alcohol extract and water extract; the concentration of the mixed solution II of the radix tetrastigme alcohol extraction and the water extraction is that each milliliter contains 0.5 g of crude drug;
(4) pouring herba Dendrobii extract, Glycyrrhrizae radix extract and radix Apioris Fortunei alcohol extraction + water extraction mixed solution II into a clean glass bottle, sealing with a gland, sterilizing at 121 deg.C for 20-30min, and cooling to obtain antiviral Chinese medicinal composition.
Pharmacodynamic test
Sample preparation: the traditional Chinese medicine compositions prepared in examples 2, 4, 6 and 8 were treated, and the positive control drugs were: ribavirin (Riba), entecavir, 2 'CMC (2' -C-methylcytidine);
processing a detected sample:
sample No. 1: taking the traditional Chinese medicine composition in the embodiment 2, heating in water bath at 50 ℃ for 24h, centrifuging at 12000rpm for 10min, taking supernatant (the treatment aims at removing insoluble particles so as to avoid influencing cells observed under a microscope), and reserving for later use;
sample No. 2: taking the traditional Chinese medicine composition in the embodiment 4, heating in water bath at 50 ℃ for 1h, centrifuging at 12000rpm for 10min, and taking the supernatant for later use;
sample No. 3: taking the traditional Chinese medicine composition in the embodiment 6, heating in water bath at 50 ℃ for 1h, centrifuging at 12000rpm for 10min, and taking the supernatant for later use;
sample No. 4: taking the traditional Chinese medicine composition in the embodiment 8, heating in water bath at 50 ℃ for 1h, centrifuging at 12000rpm for 10min, and taking the supernatant for later use;
evaluation of Activity of influenza Virus H1N1
Virus strain: influenza strain H1N1 PR 8;
cell model: dog kidney cell line MDCK; the culture conditions are as follows: DMEM + 10% fetal bovine serum, 37 ℃ and 5% CO2；
1. Cytotoxicity assays
The experiment adopts CellTiter-GloTM(Promega) kit for detecting the toxic effect of a sample on cells the experimental procedure was: inoculating the MDCK cells into a 96-hole cell culture plate, and keeping the cells adherent for later use; the drug was serially diluted in DMEM medium for 8 gradients of 20%, 6.67%, 2.22%, 0.74%, 0.25%, 0.08%, 0.027%, 0.009%, starting from 2 times the highest concentration tested, in 3-fold gradients; adding 100 μ L of the drug and 100 μ L of the culture medium to the cells, CO at 37 deg.C2Culturing in an incubator; after adding medicine and culturing for 24h, observing cytopathic effect (CPE) caused by the medicine under a microscope, and adding CellTiter-Glo to detect the cell survival rate; the toxicity of a drug to a cell is expressed as the activity of the cell.
Calculating the formula: cell activity (%) ═ drug group value/cell control group mean value 100
2. Antiviral Activity assay
The method comprises the following steps: inoculating MDCK cells into a 96-well cell culture plate, and culturing overnight at 37 ℃ for later use; simultaneously adding 100 mu of medicine and 100 mu of LH1N1 virus solution into MDCK cells; culturing in a 37 deg.C cell culture box for 24 hr, and collecting culture supernatant for detection.
The experiment was performed with a blank control well (normal cells), a virus control well (no drug added after virus infection), and a positive drug control well (ribavirin added after infection).
Inhibition (%) 100- (sample well number-blank number)/(virus control number-blank number) × 100
The specific inhibitory effect of the positive drug ribavirin and 4 samples on H1N1 PR8 infection and toxicity results on MDCK cells are shown in table 1 and fig. 1 and 2, wherein CC 50: a 50% cytotoxic concentration; EC (EC)50: 50% effective concentration (or 50% inhibitory concentration); SI ═ CC50/EC50。
TABLE 1 specific inhibitory Effect of ribavirin and 4 samples on H1N1 PR8 infection and toxicity results on MDCK cells
| Numbering | Storage concentration | Maximum detected concentration | Dilution factor | CC50 | EC50 | SI | |
| 1 | |
10％ | 3 | 6.11％ | 0.07％ | 87.6 | |
| 2 | |
10％ | 3 | 3.85％ | 0.08％ | 48.7 | |
| 3 | |
10％ | 3 | 4.46％ | 0.06％ | 75.52 | |
| 4 | |
10％ | 3 | 3.55％ | 0.12％ | 30.79 | |
| | 50mM | 500μM | 3 | >500 | 3.321 | 150.56 |
As can be seen from Table 1, FIG. 1 and FIG. 2, the test results show that the positive drug ribavirin has a dose-dependent inhibitory effect on influenza virus H1N1 PR8 strain by testing the virus replication level after the sample acts; the maximum detection concentration of 4 detected samples is 10%, and the samples are diluted by three times of gradient, wherein 10%, 3.33% and 1.11% of No. 1 samples are toxic, and 0.37% of samples are nontoxic; 2. samples No. 3 and No. 4 are 10% and 3.33% respectively toxic, the reagent does not detect toxicity under the concentration of 1.11%, but the samples No. 2, No. 3 and No. 4 are weak toxic to cells under the concentration of 1.11% and are non-toxic under the concentration of 0.37% through microscope observation; 1. the samples No. 2, 3 and 4 have dose-dependent inhibition effect on influenza virus H1N1 PR8 in the detected concentration range.
Evaluation of parainfluenza Virus type 3 Activity
Virus strain: parainfluenza virus HPIV 3;
cell model: monkey kidney cell line LLC-MK 2; the culture conditions are as follows: DMEM + 10% fetal bovine serum, 37 ℃ and 5% CO2；
Positive drug: ribavirin;
1. cytotoxicity assays
The experiment adopts CellTiter-GloTM(Promega) kit for detecting toxic effect of sample on cells
The experimental steps are as follows: inoculating MK2 cells into a 96-hole cell culture plate, and keeping the cells adherent; the drug was serially diluted in DMEM medium for 8 gradients of 20%, 6.67%, 2.22%, 0.74%, 0.25%, 0.08%, 0.027%, 0.009%, starting from 2 times the highest concentration tested, in 3-fold gradients; adding 100ul of the drug and 100ul of the culture medium to the cells, and adding CO at 37 deg.C2Culturing in an incubator; adding medicine and culturing for 24 hr, and observing cells induced by medicine under microscope(iii) lesion effect (CPE), cell viability assay with CellTiter-Glo; the toxicity of a drug to a cell is expressed as the activity of the cell.
Calculating the formula: cell activity (%) ═ drug group value/cell control group mean value 100
2. Antiviral Activity assay
The method comprises the following steps: MK2 cells are inoculated in a 96-hole cell culture plate and cultured overnight at 37 ℃ for later use; 100ul of medicine and 100ul of HPIV3 virus solution are added into MK2 cells at the same time; after being cultured in a cell culture box at 37 ℃ for 24h, the protein expression level is measured by an indirect immunofluorescence method.
The experiment was performed with a blank control well (normal cells), a virus control well (no drug added after virus infection), and a positive drug control well (ribavirin added after infection).
Inhibition (%) 100- (sample well number-blank number)/(virus control number-blank number) × 100
Specific monoclonal antibodies of parainfluenza virus HN protein are used for identifying HN, pictures are taken by a high content cytometric analyzer, and data analysis shows that the specific inhibition effect of the positive drug ribavirin and 4 samples on HPIV3 parainfluenza virus and the toxicity result on MK2 cells are shown in table 2, fig. 3 and fig. 4, wherein the specificity of CC 50: a 50% cytotoxic concentration; EC (EC)50: 50% effective concentration (or 50% inhibitory concentration); SI ═ CC50/EC50。
TABLE 2 ribavirin and results of toxicity on HPIV3 parainfluenza Virus specific inhibition and MK2 cells for 4 samples
| Numbering | Storage concentration | Maximum detected concentration | Dilution factor | CC50 | EC50 | SI | |
| 1 | |
10％ | 3 | ＞10％ | 0.2713％ | ＞36.5 | |
| 2 | |
10％ | 3 | ＞10％ | 0.311％ | ＞32 | |
| 3 | |
10％ | 3 | 7.291％ | 0.2933％ | 24.9 | |
| 4 | |
10％ | 3 | 4.562％ | 0.2097％ | 21.8 | |
| | 50mM | 100μM | 3 | ＞100 | 3.714 | ＞26.9 |
As can be seen from table 2, fig. 3 and fig. 4, the positive drug ribavirin has a dose-dependent inhibitory effect on parainfluenza virus HPIV3 as a result of detecting the replication level of HPIV3 after drug action on MK2 cell line; the tested sample has certain inhibition effect on the replication of parainfluenza virus HPIV3 in the MK2 cell line within the detected concentration range; the reagent test results of the sample 1 and the sample 2 at the concentration of 10% show that the reagent has no toxicity or less toxicity, but the reagent has obvious toxicity to cells through microscopic examination.
Evaluation of Activity against Hepatitis C Virus (HCV)
Virus strain: detecting the used virus strain as an infectious HCV type 2a virus strain containing an Rluc reporter gene;
cell model: detecting the used cell line as an HCV susceptible cell line hepatoma cell line Huh7.5.1; cell culture conditions: culturing in DMEM medium supplemented with 10% fetal calf serum, 100U/ml penicillin and 100g/ml streptomycin at 37 deg.C and 5% CO2Culturing in an incubator;
positive drug: 2' CMC.
1. Cytotoxicity assays
The experiment adopts CellTiter-GloTM(Promega) kit for detecting toxic effect of sample on cells
The experimental steps are as follows: inoculating Huh7.5.1 cells into a 96-hole cell culture plate, and keeping the cells adherent for later use; the drug was serially diluted in DMEM medium for 8 gradients of 20%, 6.67%, 2.22%, 0.74%, 0.25%, 0.08%, 0.027%, 0.009%, starting from 2 times the highest concentration tested, in 3-fold gradients; adding 100ul of the drug and 100ul of the culture medium to the cells, and adding CO at 37 deg.C2Culturing in an incubator; after adding medicine and culturing for 72h, observing cytopathic effect (CPE) caused by the medicine under a microscope, and adding CellTiter-Glo to detect the cell survival rate; the toxicity of a drug to a cell is expressed as the activity of the cell.
Calculating the formula: cell activity (%) ═ drug group value/cell control group mean value 100
2. Antiviral Activity assay
The method comprises the following steps: inoculating Huh7.5.1 cells into a 96-well cell culture plate for overnight culture for later use; the drug is serially diluted by 8 gradients in a DMEM medium from 2 times of the highest test concentration in a 3-fold gradient manner; adding 100ul of the medicine and 100ul of the virus solution into the cells, and culturing in a CO2 incubator at 37 ℃; after adding medicine and adding virus to culture for 72h, the Rluc reading value is detected by Renilla LuciferaseAssaykit.
Post-infection 2' CMC was a positive drug control well, cell wells without added virus and compound were cell control wells, and wells with added virus only were virus control wells.
Inhibition (%) 100- (sample well number-blank number)/(virus control number-blank number) × 100
Through detection, the specific inhibition effect of the positive drug 2' CMC and 4 samples on HCV virus and the toxicity result on huh7.5.1 cell are shown in table 3 and fig. 5, wherein the ratio of CC 50: a 50% cytotoxic concentration; EC (EC)50: 50% effective concentration (or 50% inhibitory concentration); SI ═ CC50/EC50。
TABLE 2 positive drug 2' CMC and 4 samples specific inhibition of HCV virus and toxicity results on Huh7.5.1 cells
| Numbering | Storage concentration | Maximum detected concentration | Dilution factor | CC50 | EC50 | SI |
| 1 | |
10％ | 3 | 3.423％ | 0.05764％ | 59.4 |
| 2 | |
10％ | 3 | 3.648％ | 0.04718％ | 77.3 |
| 3 | |
10％ | 3 | 3.679％ | 0.05518％ | 66.7 |
| 4 | |
10％ | 3 | 3.187％ | 0.04709％ | 67.7 |
| 2’CMC | 50mM | 100μM | 3 | ＞30 | 1.751 | ＞17 |
As can be seen from table 3 and fig. 5, the positive drug 2' CMC has a dose-dependent inhibitory effect on HCV type 2a virus as a result of detecting the replication level of HCV infection after drug action on huh7.5.1 cell line. The tested sample has certain inhibiting effect on the replication of HCV 2a in the Huh7.5.1 cell line within the tested concentration range.
Evaluation of Activity against Hepatitis B Virus (HBV)
Cells and cell models: HepG 2-derived HBV genome stably transfected cell line HepAD 38; the culture conditions are as follows: DMEM/F12+ 10% fetal bovine serum, 37 ℃, 5% CO2；
Positive drug: entecavir;
1. cytotoxicity assays
Detecting the toxic effect of the drug on cells by DAPI staining;
the experimental steps are as follows: inoculating HepG 2-derived HBV genome stably transfected cells HepAD38 into a 96-hole cell culture plate, and keeping the cells adherent for later use; the drug was serially diluted in maintenance medium for 8 gradients of 20%, 6.67%, 2.22%, 0.74%, 0.25%, 0.08%, 0.027%, 0.009%, starting from 2 times the highest concentration tested, in 3-fold gradients; adding 100lu of the drug and 100ul of culture medium to the cells, CO at 37 deg.C2Culturing in an incubator; after adding medicine and culturing for 3 days, detecting the cell survival rate by DAPI staining;
calculating the formula: cell activity (%) ═ drug group value/cell control group mean value 100
2. Activity assay for HBV antigens
The method comprises the following steps: cells with Dox removed are inoculated in a 96-well cell culture plate, 100 mu L of medicine and 100 mu L of culture medium are added after the cells grow into a monolayer, culture is carried out at 37 ℃ for 72h, culture supernatant is taken, and an ELISA kit is used for detecting the expression content of HBeAg and HBsAg.
The experiment was performed with a cell control well (cell well without virus and compound) and a positive drug (entecavir after infection) control well.
Inhibition (%) - (absorbance of cell control-drug)/(absorbance of cell control-blank) 100%
3. Activity assay for HBVDNA
The method comprises the following steps: inoculating cells with Dox removed into a 96-well cell culture plate, adding 200ul of maintenance culture medium containing various medicines after the cells grow into a single layer, culturing at 37 ℃ for 72h, removing supernatant, washing the cells twice with PBS, and adding lysis solution to lyse the cells; extracting total DNA of cells by phenol chloroform, precipitating the DNA by ethanol, and dissolving the DNA by TE; HBV DNA and GAPDH were quantified by fluorescence quantification.
The experiment was set with a virus control well (virus only wells) and a positive drug (entecavir after infection) control well.
Inhibition (%). 100-drug/virus control value 100
The results of the inhibition of entecavir and samples nos. 1, 2, 3 and 4 on HBeAg, HBsAg and DNA of HBV and the toxicity on HepAD38 cell are shown in table 4, fig. 6, fig. 7 and fig. 8, wherein CC 50: a 50% cytotoxic concentration; EC (EC)50: 50% effective concentration (or 50% inhibitory concentration); SI ═ CC50/EC50(ii) a "-": no virus inhibitory activity was detected, no selection index.
TABLE 4 results of Entecavir inhibition and cell toxicity of samples No. 1, 2, 3, and 4 against HBeAg, HBsAg, and DNA of HBV
As can be seen from table 4, fig. 6, fig. 7 and fig. 8, the results of detecting the HBeAg, HBsAg and DNA levels of the HBV virus show that the positive drug entecavir has no inhibitory effect on the expression of HBeAg and HBsAg, and has a strong inhibitory effect on HBV DNA; the detected sample of the traditional Chinese medicine composition No. 1, 2, 3 and 4 has obvious drug insoluble substances at the highest detection concentration of 10 percent, and compared with the cells of a control group, the detected sample of the traditional Chinese medicine composition No. 1, 2, 3 and 4 obviously inhibits the cell proliferation of HepAD38 at the concentrations of 10 percent and 3.3 percent, changes the cell form (cell rounding) and has obvious toxic effect on the cells; 1. the CC50 toxicity of samples No. 2, 3, and 4 to HepAD38 cells was > 10%, and 5.23%, respectively; EC50 for HBeAg was 3.737%, 3.309%, 3.112% and 3.241%, respectively; EC50 for HBsAg was 1.904%, > 10%, 3.784% and 2.496%, respectively; has no inhibition effect on HBV DNA replication; the antiviral effect of the Chinese medicinal composition on the antigens HBeAg and HBsAg of HBV can be caused by cytotoxicity.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. An antiviral traditional Chinese medicine composition is characterized by comprising the following raw materials: 2-3 parts of radix tetrastigme, 1 part of dendrobium officinale and 1 part of liquorice.
2. The antiviral traditional Chinese medicine composition as claimed in claim 1, which specifically comprises the following raw materials: the traditional Chinese medicine composition comprises, by weight, 2 parts of radix tetrastigme, 1 part of dendrobium officinale and 1 part of liquorice.
3. The preparation method of the antiviral traditional Chinese medicine composition is characterized by comprising the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing radix tetrastigme ultrafine powder: weighing 2-3 parts of raw material radix tetrastigme, and crushing into superfine powder for later use;
(4) sterilizing the dendrobium officinale extract, the liquorice extract and the radix tetrastigme ultrafine powder respectively, and mixing uniformly to obtain the antiviral traditional Chinese medicine composition.
4. The preparation method of the antiviral traditional Chinese medicine composition is characterized by comprising the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing an ethanol extract of radix tetrastigme: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40-70% ethanol, boiling, reflux extracting for 2-4h for 3 times, filtering, mixing filtrates, and concentrating to obtain radix tetrastigme ethanol extract for use;
(4) and uniformly mixing the dendrobium officinale extract, the liquorice extract and the radix tetrastigme alcohol extract, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
5. The preparation method of the antiviral traditional Chinese medicine composition is characterized by comprising the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing a mixed solution I of radix tetrastigme alcohol extraction and water extraction: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40-70% ethanol, boiling, reflux extracting for 2-4h for 3 times, filtering, collecting residue, mixing filtrates, and concentrating to obtain radix tetrastigme ethanol extract; adding water into the residue, boiling, reflux-extracting for 2-4 hr for 3 times, filtering, mixing filtrates, and concentrating to obtain radix Apioris Fortunei water extractive solution; mixing the radix tetrastigme alcohol extract and the radix tetrastigme water extract to obtain a radix tetrastigme alcohol extraction and water extraction mixed solution I;
(4) and uniformly mixing the dendrobium officinale extracting solution, the liquorice extracting solution and the radix tetrastigme alcohol extraction and water extraction mixed solution I, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
6. The preparation method of the antiviral traditional Chinese medicine composition is characterized by comprising the following steps:
(1) preparing a dendrobium officinale extracting solution: weighing 1 part of raw material dendrobium officinale, adding water, boiling, refluxing and extracting for 4-8h for 3 times, then filtering, combining filtrates, and concentrating to obtain dendrobium officinale extract for later use;
(2) preparing a liquorice extracting solution: weighing 1 part of raw material liquorice, adding water, boiling, refluxing and extracting for 1-3h for 3 times, then filtering, combining filtrates, and concentrating to obtain liquorice extract for later use;
(3) preparing a mixed solution II of radix tetrastigme alcohol extraction and water extraction: weighing 2-3 parts of radix tetrastigme as raw material, adding 5-15 times of 40-70% ethanol, boiling, reflux extracting for 2-4h for 3 times, filtering, reserving filter residue, and combining filtrates to obtain radix tetrastigme ethanol extract; adding water into the residue, boiling, reflux-extracting for 2-4 hr for 3 times, filtering, and mixing filtrates to obtain radix Apioris Fortunei water extractive solution; mixing the radix tetrastigme alcohol extract and the radix tetrastigme water extract, refrigerating overnight, centrifuging, and filtering to obtain a radix tetrastigme alcohol extraction and water extraction mixed solution II;
(4) and uniformly mixing the dendrobium officinale extracting solution, the liquorice extracting solution and the radix tetrastigme alcohol extraction and water extraction mixed solution II, and sterilizing to obtain the antiviral traditional Chinese medicine composition.
7. The method for preparing an antiviral Chinese medicinal composition according to any one of claims 3 to 6, wherein the volume of the water in step (1) is 15 to 30 times of the mass of the Dendrobium officinale; the volume of the water in the step (2) is 10-15 times of the mass of the liquorice.
8. The method for preparing an antiviral traditional Chinese medicine composition according to claim 3, wherein the sterilization conditions of the dendrobium officinale extract and the licorice extract in the step (4) are as follows: sterilizing at 121 deg.C for 20-30 min; the radix tetrastigme ultrafine powder is sterilized by irradiating with 25-50KGY cobalt 60-gamma rays for 18-22 min.
9. The method for preparing an antiviral Chinese medicinal composition according to any one of claims 4 to 6, wherein the sterilization conditions in step (4) are all sterilization at 121 ℃ for 20 to 30 min.
10. The use of a Chinese medicinal composition as claimed in claim 1 or 2 or prepared by the preparation method as claimed in any one of claims 3 to 9 in an antiviral medicament, wherein the virus is influenza virus, parainfluenza virus, hepatitis b virus and hepatitis c virus.
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