CN112451576A - Anti-coronavirus composition, preparation method and application thereof - Google Patents
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Abstract
The invention relates to an anti-coronavirus composition, a preparation method and application thereof, belonging to the technical field of medicines. The invention discloses an application of an anti-coronavirus composition in preparing a medicament for preventing and/or treating diseases caused by coronavirus, wherein the anti-coronavirus composition comprises the following components in parts by weight: 5-100 parts of roxburgh rose, 5-100 parts of emblic leafflower fruit, 5-100 parts of honey, 1-30 parts of walnut and 1-30 parts of lemon. The anti-coronavirus composition of the present invention has excellent anti-SARS-CoV-2 activity.
Description
Technical Field
The invention relates to an anti-coronavirus composition, a preparation method and application thereof, belonging to the technical field of medicines.
Background
The new coronary pneumonia (COVID-19) caused by the novel coronavirus (SARS-CoV-2) is a serious public health event, and the outbreak has caused huge casualties and economic losses so far, but at present, no specific medicine aiming at the SARS-CoV-2 with definite curative effect exists clinically. In addition, most patients are treated by antibiotics, interferons, oxygen supply, glucocorticoids, traditional Chinese medicines and other medicines, but the medicines are not novel specific medicines for coronavirus. At present, part of potential antiviral drugs pass through a rapid review channel, but clinical tests are not finished yet and the effectiveness is to be determined.
In 2020, 10 and 22 days, the antiviral drug Reidesvir (trade name: Veklury) approved by the US FDA is sold on the market, and is suitable for treating patients with new crown pneumonia which is 12 years old or more, has the weight of at least 40 kg and needs hospitalization. In 5 months of 2020, the FDA grants redciclovir emergency use authorization, allowing medical personnel to use hospitalized patients with new coronary pneumonia. However, the efficacy of Reidesciclovir is controversial. The world health organization publishes the clinical test results of the medicines such as the Reidesciclovir and the hydroxychloroquine in 10 months and 15 days, and the medicines are said to be 'almost ineffective' for reducing the fatality rate of the new coronary pneumonia. Therefore, the development of novel antiviral drugs having proprietary intellectual property rights is urgently required.
Patent application CN201710226412.0 discloses a Chinese herbal compound capable of improving body functions, which is prepared by taking the traditional medical theory of the Yi nationality as guidance, crushing or slicing roxburgh rose, emblic leafflower fruit, honey, walnut and lemon, extracting with alcohol, percolating, collecting extract or percolate, adding honey, mixing uniformly, and preparing into oral preparation; or fermenting or soaking the above medicinal materials or food materials to obtain oral preparation. The invention uses the compound fermentation liquor preparation to process new coronavirus, and finds that the compound fermentation liquor preparation has excellent anti-SARS-CoV-2 activity.
Disclosure of Invention
The first object of the present invention is to provide a new use of the traditional Chinese medicine composition disclosed in CN201710226412.0 in preparing anti-coronary virus drugs.
To achieve the first object of the present invention, an anti-coronavirus composition for preventing and/or treating a disease caused by coronavirus comprises the following components in parts by weight: 5-100 parts of roxburgh rose, 5-100 parts of emblic leafflower fruit, 5-100 parts of honey, 1-30 parts of walnut and 1-30 parts of lemon.
In one embodiment, the anti-coronavirus composition comprises: the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
In one embodiment, the anti-coronavirus composition is prepared by a method comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
In one embodiment, the method comprises: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
In a specific embodiment, the coronavirus comprises a novel coronavirus SARS-CoV-2, Severe acute respiratory syndrome SARS, middle east respiratory syndrome MERS, or human enterocoronavirus.
In one embodiment, the coronavirus comprises the novel coronavirus SARS-CoV-2.
It is a second object of the present invention to provide an anti-coronavirus composition, said composition comprising: fructus Rosae Normalis, fructus Phyllanthi, Mel, semen Juglandis, and fructus Citri Limoniae; the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
In one embodiment, the composition is prepared by a method comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
In one embodiment, the method comprises:
mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
The third object of the present invention is to provide a method for preparing the above-mentioned anti-coronavirus composition.
To achieve the third object of the present invention, the method comprises: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
Has the advantages that:
the anti-coronavirus drug of the invention has excellent anti-SARS-CoV-2 activity.
Detailed Description
To achieve the first object of the present invention, an anti-coronavirus composition for preventing and/or treating a disease caused by coronavirus comprises the following components in parts by weight: 5-100 parts of roxburgh rose, 5-100 parts of emblic leafflower fruit, 5-100 parts of honey, 1-30 parts of walnut and 1-30 parts of lemon.
In one embodiment, the anti-coronavirus composition comprises: the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
In one embodiment, the anti-coronavirus composition is prepared by a method comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
In one embodiment, the method comprises: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
In a specific embodiment, the coronavirus comprises a novel coronavirus SARS-CoV-2, Severe acute respiratory syndrome SARS, middle east respiratory syndrome MERS, or human enterocoronavirus.
In one embodiment, the coronavirus comprises the novel coronavirus SARS-CoV-2.
It is a second object of the present invention to provide an anti-coronavirus composition, said composition comprising: fructus Rosae Normalis, fructus Phyllanthi, Mel, semen Juglandis, and fructus Citri Limoniae; the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
In one embodiment, the composition is prepared by a method comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
In one embodiment, the method comprises:
mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
The third object of the present invention is to provide a method for preparing the above-mentioned anti-coronavirus composition.
To achieve the third object of the present invention, the method comprises: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
The following examples are provided to further illustrate the embodiments of the present invention and are not intended to limit the scope of the present invention.
Example 1
Taking 400g of roxburgh rose, 400g of emblic leafflower fruit, 400g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at the room temperature of 15-40 ℃ in a shade, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Example 2
Taking 200g of roxburgh rose, 150g of emblic leafflower fruit, 250g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Example 3
Taking 300g of roxburgh rose, 400g of emblic leafflower fruit, 500g of honey, 150g of walnut and 150g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Example 4
Taking 400g of roxburgh rose, 250g of emblic leafflower fruit, 200g of honey, 75g of walnut and 75g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Comparative example 1
Taking 1200g of roxburgh rose, 400g of emblic leafflower fruit, 400g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Comparative example 2
Taking 400g of roxburgh rose, 1200g of emblic leafflower fruit, 400g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the components, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Comparative example 3
Taking 400g of roxburgh rose, 400g of emblic leafflower fruit, 400g of honey, 400g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the components, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Comparative example 4
Taking 400g of emblic leafflower fruit, 400g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the phyllanthus emblic leafflower fruit tea.
Comparative example 5
Taking 400g of roxburgh rose, 400g of honey, 100g of walnut and 100g of lemon, adding water which is 4 times of the total weight of the formula, fermenting for 30 days in a dark place at 15-40 ℃, sealing, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the roxburgh rose beverage.
Comparative example 6
Taking 400g of roxburgh rose, 400g of emblic leafflower fruit, 400g of honey and 100g of walnut, adding water which is 4 times of the total weight of the formula, fermenting for 30 days in a dark place at 15-40 ℃, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the tea.
Comparative example 7
Taking 400g of emblic leafflower fruit, adding water which is 4 times of the total weight of the formula, fermenting for 30 days at 15-40 ℃ in a dark place, filtering, collecting fermentation liquor, and subpackaging into 100 ml/bottle to obtain the phyllanthus emblic leafflower fruit fermented beverage.
Test example 1
Research on the coronavirus resistance of fermentation liquor.
(1) Cytotoxicity test
Vero E6 cells frozen in a liquid nitrogen tank were removed and revived according to standard procedures. After the cells are passed twice, taking out pairsAfter the cells were attached to the wall, the fermentation liquids in each example and comparative example were diluted to a certain concentration with the medium, added to the cells, and cultured for 72 hours. Adding 20 microliters of MTT (5mg/ml) into each hole, absorbing the liquid by using a discharging gun after the action for 4 hours, adding 150 microliters of DMSO into each hole, oscillating the mixture in a shaking table for about 10 minutes, and measuring the absorbance value of 570 by using a microplate reader; calculating the inhibition rate according to the absorbance of the data, and then calculating the half cytotoxic concentration CC50。
(2) Plaque Reduction Neutralization Test (PRNT)
1) Vero E6 cells, 20 ten thousand cells per well in 24-well plates, were cultured overnight to allow attachment.
2) Each serum sample was diluted stepwise in 2-fold gradient with DMEM medium containing 2% inactivated FBS, starting at 5-fold dilution, for a total of 6 concentration gradients, i.e., 5, 10, 20, 40, 80, 160. The volume of each diluted sample was 125. mu.L. The specific method comprises the following steps: firstly, 36 serum samples (6 immunization groups multiplied by 6 animals/group) are centrifuged, and the liquid flows to the bottom of an EP tube; adding 200 mu L of DMEM medium containing 2% inactivated FBS into a centrifuge tube containing 50 mu L of serum, and uniformly mixing to obtain a sample diluted by 5 times; taking out 125 mu L of the solution, and adding the solution into 125 mu L of DMEM medium containing 2% inactivated FBS to obtain a sample diluted by 10 times; and by analogy, gradually diluting to obtain samples diluted by 20 times, 40 times, 80 times and 160 times respectively.
3) SARS-Cov-2 virus was diluted to 400PFU/mL with DMEM medium containing 2% inactivated FBS.
4) Mixing the diluted serum sample with SARS-Cov-2 virus in equal volume, and incubating for 1 hour at 37 ℃; meanwhile, DMEM medium containing 2% inactivated FBS is used as a blank control group, mixed with SARS-Cov-2 virus in equal volume, subjected to parallel test, and incubated at 37 ℃ for 1 hour.
5) The Vero cell medium was discarded, and the heat-incubated virus-serum mixture was added to the cells at 200. mu.L per well.
6) The incubator was incubated for 1 hour, during which the plates were shaken every 15 minutes.
7) After 1 hour the virus-containing medium was discarded and washed once with PBS.
8) The low melting point agarose 5% in concentration was melted by heating at 65-100 degrees and diluted to 1% concentration with 37 degrees pre-warmed DMEM medium containing 2% inactivated FBS when it was cooled to be slightly non-scalding (about 40-60 degrees). Then 250. mu.L per well (24 well plate in step 7) was added.
9) The incubator was incubated for 48 hours.
10) The medium was discarded, fixed with 4% paraformaldehyde for 15 minutes, and then stained with 0.2% crystal violet.
11) Counting the number of plaques
Calculating the half effective concentration EC according to the number of the plaques50。
Selection index SI ═ CC50/EC50. The results of the in vitro experiments are shown in table 1.
TABLE 1 study of the activity of fermentation broths against neocoronaviruses
| Half cytotoxic concentration CC50(％) | Half the effective concentration EC50(％) | Selection index SI | |
| Example 1 | 5.55 | 0.13 | 42.69 |
| Example 2 | 6.74 | 1.25 | 5.39 |
| Example 3 | 8.17 | 0.96 | 8.51 |
| Example 4 | 7.25 | 1.38 | 5.25 |
| Comparative example 1 | 9.14 | 2.37 | 3.86 |
| Comparative example 2 | 10.23 | 5.32 | 1.92 |
| Comparative example 3 | 6.94 | 2.14 | 3.24 |
| Comparative example 4 | 5.73 | 2.58 | 2.22 |
| Comparative example 5 | 6.19 | 1.46 | 4.24 |
| Comparative example 6 | 7.35 | 2.24 | 3.28 |
| Comparative example 7 | 11.42 | 3.18 | 3.59 |
Note: the values of the median cytotoxic concentration and median effective concentration in the table are the percentage of the medium diluted with broth; the plaque reduction neutralization experiment is carried out under the condition that the fermentation liquor has no cytotoxicity; selection index SI ═ CC50/EC50。
The results of the cytotoxicity test and plaque reduction neutralization test are shown in Table 1. As can be seen from the table, the selection index of examples 1-4 is 5.25-42.69, which is higher than that of comparative examples 1.92-4.24, wherein the half inhibitory concentration CC of the fermentation liquid of example 1 on Vero E6 cells505.55%, half effective concentration EC for SARS-CoV-250The index was chosen to be 42.69 at 0.13%, whereas in all other comparative examples the index was chosen to be much lower than in example 1. It was shown that the fermentation broth of example 1 had excellent anti-SARS-CoV-2 activity.
Claims (10)
1. Use of an anti-coronavirus composition in the preparation of a medicament for preventing and/or treating a disease caused by a coronavirus, wherein the anti-coronavirus composition comprises the following components in parts by weight: 5-100 parts of roxburgh rose, 5-100 parts of emblic leafflower fruit, 5-100 parts of honey, 1-30 parts of walnut and 1-30 parts of lemon.
2. Use according to claim 1, characterized in that said anti-coronavirus composition comprises: the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
3. Use according to claim 1 or 2, characterized in that said anti-coronavirus composition is prepared by a process comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
4. Use according to claim 3, characterized in that the method comprises: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
5. Use according to any one of claims 1 to 4, wherein the coronavirus is a novel coronavirus, severe acute respiratory syndrome, middle east respiratory syndrome or human enterocoronavirus.
6. Use according to claim 5, wherein the coronavirus is a novel coronavirus.
7. An anti-coronavirus composition, comprising: fructus Rosae Normalis, fructus Phyllanthi, Mel, semen Juglandis, and fructus Citri Limoniae; the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon are in the following weight ratio: 3-4: 4:4:1:1, preferably 4:4:4:1: 1.
8. The anti-coronavirus composition of claim 7, wherein the composition is prepared by a method comprising: and (3) fermenting the roxburgh rose, the phyllanthus emblica, the honey, the walnut and the lemon.
9. The anti-coronavirus composition of claim 8, wherein the method comprises:
mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
10. A process for the preparation of an anti-coronavirus composition according to claim 9, said process comprising: mixing the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon with water, and fermenting for 30-300 days at 15-40 ℃ in a dark place under a sealed condition;
the water accounts for 2-6 times of the total amount of the roxburgh rose, the emblic leafflower fruit, the honey, the walnut and the lemon, and is preferably 3-5 times.
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| CN115844962A (en) * | 2022-09-29 | 2023-03-28 | 贵州医科大学 | Application of Rosa roxburghii extract in preparation of preparation for preventing or treating novel coronavirus infection |
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| CN102972814A (en) * | 2011-09-07 | 2013-03-20 | 米兴才 | Roxburgh rose juice drink |
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| Title |
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| "核桃粉生产技术 ", 《农村.农业.农民(B版)》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115844962A (en) * | 2022-09-29 | 2023-03-28 | 贵州医科大学 | Application of Rosa roxburghii extract in preparation of preparation for preventing or treating novel coronavirus infection |
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