CN111714596A

CN111714596A - A functional plant extract with respiratory virus resisting effect

Info

Publication number: CN111714596A
Application number: CN202010681495.4A
Authority: CN
Inventors: 吴杰军; 陈景才; 余海兰
Original assignee: Biyu Pharmaceutical Co ltd
Current assignee: Biyu Pharmaceutical Co ltd
Priority date: 2020-07-15
Filing date: 2020-07-15
Publication date: 2020-09-29
Anticipated expiration: 2040-07-15
Also published as: CN111714596B

Abstract

The invention relates to a functional plant extract with respiratory virus resistance, which can effectively prevent respiratory infection and relieve symptoms after infection, and the technical scheme is that the functional plant extract is prepared from the following raw materials in parts by weight: the traditional Chinese medicine decoction preparation is prepared from the following raw materials, by weight, 1-30 parts of honeysuckle, 1-30 parts of chrysanthemum, 1-30 parts of dandelion, 1-30 parts of astragalus membranaceus, 1-30 parts of schizonepeta, 1-30 parts of liquorice, 1-30 parts of pogostemon cablin, 1-30 parts of fried semen coicis, 1-30 parts of Chinese dates, 1-30 parts of mint and 0.001-0.1 part of sucralose.

Description

A functional plant extract with respiratory virus resisting effect

Technical Field

The invention relates to the technical field of medicines and foods, in particular to a functional plant extract with respiratory virus resistance.

Background

The respiratory viruses mainly comprise rhinoviruses, coronaviruses, enteroviruses, adenoviruses and respiratory syncytial viruses; viral upper respiratory tract infection caused by respiratory tract virus is common disease and frequently-occurring disease in the period of children, can occur all the year round, can occur several times per year for each person, is particularly most susceptible to infection in winter, and is caused by pathogen mainly invading nose, pharynx, tonsil and throat. Acute upper respiratory infection caused by various respiratory viruses, including common cold, can occur 1-3 times per year in adults, viruses can invade different parts of the upper respiratory tract to cause inflammation, and the upper respiratory infection is often combined with bacterial infection to cause exacerbation of the disease. Symptoms after respiratory virus infection comprise common cold type, the main disease season is mostly in late summer and early autumn, the disease can be continued to the next spring, the symptoms are pharyngeal discomfort or pharyngalgia, followed by watery nasal discharge, nasal obstruction, sneezing, cough, limb ache, hypodynamia and headache, sometimes accompanied by fever of different degrees, leucocyte counting weight is mostly reduced or normal, the disease course can be self-healed in 3-7 days, but the disease can also last for more than weeks; and upper respiratory tract infection of pharyngitis type, the main symptoms are pharyngalgia, watery nasal discharge, nasal obstruction, headache, cough and general malaise, red and swollen pharynx, amygdala lymphoproliferation, little exudate, swelling and tenderness of mandibular lymph node, low or normal leucocyte count, and self-healing after 3-7 days.

In the last two decades, emerging pneumonia caused by coronaviruses has led to infection and death in a large number of people, including atypical pneumonia SARS, which occurs in china in 2003, middle east respiratory syndrome MERS, which occurs in the middle east in 2012, and the global pandemic new coronaic pneumonia nCOV-19 beginning in 2019. To date, there is a lack of specific drug and vaccine treatments and prophylaxis of such emerging coronaviruses.

Traditional Chinese medicine has a history of thousands of years and plays an important role in the struggle against pestilence all the time. Therefore, a new plant extract which is used for resisting respiratory viruses and takes medicinal and edible medicinal materials as raw materials is imperative to be found according to the theoretical basis of the traditional Chinese medicine epidemic febrile disease, the classical famous formula and the folk proved formula.

Disclosure of Invention

In view of the above situation, the present invention aims to provide a functional plant extract with respiratory virus resistance, which can effectively prevent respiratory infection and alleviate symptoms after infection.

The technical scheme of the invention is that the functional plant extract is prepared from the following raw materials in parts by weight: 1-30 parts of honeysuckle, 1-30 parts of chrysanthemum, 1-30 parts of dandelion, 1-30 parts of astragalus, 1-30 parts of schizonepeta, 1-30 parts of liquorice, 1-30 parts of pogostemon cablin, 1-30 parts of fried semen coicis, 1-30 parts of Chinese date, 1-30 parts of mint and 0.001-0.1 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus, the liquorice, the fried semen coicis and the Chinese date together, adding 10-250 times of purified water for soaking for 30-60 minutes, then boiling for 10-60 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuing to decoct for 10-60 minutes, after clarifying and cooling to room temperature (23 +/-2 ℃), adding the sucralose, filtering, subpackaging and sterilizing, thus obtaining the health-care.

The honeysuckle, the chrysanthemum, the dandelion, the astragalus, the schizonepeta, the liquorice, the patchouli, the fried coix seed, the Chinese date and the mint meet the requirements of the latest national pharmacopoeia standard.

The invention can also be prepared into beverages, liquid and solid preparations according to the mature process technology.

The traditional Chinese medicine decoction technology is adopted, active ingredients of medicinal materials are kept to the maximum extent, respiratory viruses can be effectively prevented from entering cells, and replication of the viruses in the cells is inhibited, and the traditional Chinese medicine decoction can be used as a plant extract with homology of medicine and food.

Detailed Description

The following examples further illustrate the embodiments of the present invention in detail.

Example 1

The invention can be prepared from the following raw materials in parts by weight in specific implementation: 10 parts of honeysuckle, 10 parts of chrysanthemum, 10 parts of dandelion, 10 parts of astragalus membranaceus, 10 parts of schizonepeta, 10 parts of liquorice, 10 parts of pogostemon cablin, 10 parts of fried semen coicis, 10 parts of Chinese date, 10 parts of mint and 0.02 part of sucralose, wherein the honeysuckle, the chrysanthemum, the dandelion, the astragalus membranaceus, the liquorice, the fried semen coicis and the Chinese date are mixed together, 10 times of purified water is added for soaking for 30 minutes, then the mixture is boiled for 30 minutes, the schizonepeta, the pogostemon cablin and the mint are added, the decoction is continued for 30 minutes, the sucralose is added after the mixture is clarified and cooled to the.

Example 2

When the invention is implemented, the invention can also be prepared by the following raw materials by weight: 20 parts of honeysuckle, 20 parts of chrysanthemum, 20 parts of dandelion, 20 parts of astragalus membranaceus, 20 parts of schizonepeta, 20 parts of liquorice, 20 parts of pogostemon cablin, 20 parts of fried semen coicis, 20 parts of Chinese date, 20 parts of mint and 0.05 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus membranaceus, the liquorice, the fried semen coicis and the Chinese date together, adding 50 times of purified water, soaking for 30 minutes, then boiling for 30 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuously decocting for 30 minutes, adding the sucralose after clarifying and cooling to room temperature, filtering, subpackaging and sterilizing to obtain the.

For the convenience of the subsequent experiments, the product of example 2 was named "KH-0016".

Example 3

When the invention is implemented, the invention can also be prepared by the following raw materials by weight: 30 parts of honeysuckle, 30 parts of chrysanthemum, 30 parts of dandelion, 30 parts of astragalus, 30 parts of schizonepeta, 30 parts of liquorice, 30 parts of pogostemon cablin, 30 parts of fried semen coicis, 30 parts of Chinese date, 30 parts of mint and 0.1 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus, the liquorice, the fried semen coicis and the Chinese date together, adding 250 times of purified water, soaking for 30 minutes, then boiling for 30 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuing to decoct for 30 minutes, adding the sucralose after clarifying and cooling to room temperature, filtering, subpackaging and sterilizing to obtain the traditional Chinese medicine.

In the formula, the honeysuckle, the chrysanthemum, the dandelion, the schizonepeta and the mint have the functions of clearing away heat and toxic materials, dispersing wind-cold, dispelling wind and removing dampness, promoting urination and removing stasis, regulating qi and relieving epigastric distention, and resisting inflammation and disinfection; herba Agastaches has effects of eliminating turbid pathogen with aromatics, promoting appetite, relieving vomit, relieving exterior syndrome and relieving summer-heat, and radix astragali has effects of invigorating qi, consolidating exterior, invigorating qi and tonifying deficiency; the parched Coicis semen has effects of invigorating spleen and promoting diuresis; the Chinese dates can tonify middle-jiao and Qi, nourish blood and soothe nerves; the liquorice has the effects of eliminating phlegm, relieving cough and conditioning various medicines. The prescription of the invention is reasonably compatible, thereby achieving the purpose of resisting respiratory viruses without hurting spleen and stomach, and the beneficial effects are fully proved by cytological experiments, and the related experimental data are as follows:

1. the function of the functional plant extract of the invention for resisting respiratory syncytial virus

1.1 Experimental methods:

the reference compound, namely the lomitecitabine, in the detection experiment of the respiratory syncytial virus is prepared into 10 mM which is used as the highest concentration of the experiment to be stored in a refrigerator at the temperature of 20 ℃ below zero for standby, 10 XDPBS is added into KH-0016 to adjust the osmotic pressure to 1 XDPBS, and then saturated sodium bicarbonate solution at the temperature of 37 ℃ is used for adjusting the pH to 7.0-7.2. HEp2 cells were cultured in complete medium DMEM/F12 (10% FBS +1% P/S) and cultured as 1: 4, maintaining passage. HEp2 cells were added to 96-well cell culture plates 5,000 cells/well the day before the experiment. On the day of the experiment, the medium in the cell culture plate was discarded, and 100 μ L/well of diluted reference compound and KH-0016 were added. The respiratory syncytial virus stock was thawed on ice and made up to approximately 4,000 fold virus dilution in assay medium containing 2% FBS and mixed well. 100 muL of virus diluent is added into each well of the cell detection plate, and finally the virus is diluted by 8,000 times in each well of 200 muL. The Cell culture plate was cultured in an incubator at 37 ℃ and 5% CO2 for 5 days, and then the Cell activity was measured using CellTiter Cell Viability Kit (product of Promega). To evaluate the cytotoxic toxicity of the reference compound (lomixitabine) and KH-0016, a cytotoxicity control experiment was set up in the same manner as above, except that no virus was added.

1.2 Experimental results:

TABLE 1 results of anti-respiratory syncytial virus experiments (EC 50)

Compound (I)	EC50
		Rumicita (a Chinese character of' LumiBin (mu M)	21.62
KH-0016（mg/ml）	0.1678

TABLE 2 cytotoxic assay results of HEp2 cells with test substances (CC50)

Compound (I)	CC50
		Rumi Xita Bin (mu M)	>200
KH-0016（mg/ml）	>9.1

The result shows that the functional plant extract has good effect of resisting respiratory syncytial virus, and the selection index SI is greater than 54.2.

2. The function of the functional plant extract of the invention for resisting influenza virus

2.1 Experimental methods:

in this experiment, the reference compound Zanamivir (Zanamivir, Selleck, Cat S3007) was stored at 30 mM. KH-0016 was added with 10 XDPBS to adjust the osmotic pressure to 1 XDPBS, and then adjusted to pH 7.0-7.2 with saturated sodium bicarbonate solution at 37 ℃. MDCK cells were cultured in complete medium DMEM (10% FBS +1% P/S) and maintained at 1:8 for passaging. MDCK cells were added to a 96-well experimental cell plate one day before the experiment, and the cell density was about 90% on the day of the experiment at 9,000 cells per well (200 μ L MDCK cell complete medium). The complete cell culture medium of 96-well experimental cell plates was discarded, and 200 μ L of complete DMEM medium (cddmem) per well was added to wash the cells. The medium was discarded, and 200 μ L of complete DMEM medium per well (cddmem) was added to repeat the washing once. Stock influenza virus was thawed rapidly in cold water and diluted 20 ten thousand fold with influenza virus medium, approximately 200 TCID 50/mL. The culture medium of the 96-well experimental cell plate is discarded, the prepared compound working plate is taken, and 100 muL/well is added into the cell culture detection plate. Then 100 μ L of influenza virus (20 TCID 50/well) was added per well. 200 muL of influenza virus culture medium is added into ZPE (0% action) holes, 100 muL of influenza virus culture medium is added into HPE (100% action) holes, and 100 muL of influenza virus is added, and about 20TCID50 is obtained. Incubating in a 5% CO2 incubator at 37 ℃ for 72 hours; after 72 hours incubation, the cell activity was measured using the Promega CellTiter CellViability kit.

To evaluate the cytotoxic toxicity of the reference compound (zanamivir) and KH-0016, a cytotoxicity control experiment was set up, as above, but without the addition of virus.

2.2 Experimental results:

TABLE 3 anti-influenza Virus test results (EC 50)

Compound (I)	EC50
		Zanamivir (mu M)	18.61
KH-0016（mg/ml）	0.82

TABLE 4 cytotoxic assay results of MDCK cells with test substances (CC50)

Compound (I)	CC50
		Zanamivir (mu M)	>100
KH-0016（mg/ml）	>5.7

The result shows that the functional plant extract has good function of resisting influenza virus. The selection index SI > 6.9.

3. The functional plant extract of the invention has the effect of resisting coronavirus

3.1 Experimental methods:

in this experiment, the reference compound Ribavirin (Ribavirin) was stored at 30 mM and KH-0016 was added to 10 × DPBS to adjust the osmotic pressure to 1 × DPBS, and then 37 was usedpH was adjusted to 7.0-7.2 with saturated sodium bicarbonate solution the experiment was performed on passaged Huh7 cells in (DMEM +10% FBS +1% P/S) at 1.5 × 10⁴One/well was inoculated in a 96-well plate, and 100 TCID was added after overnight culture₅₀HCoV-229E virus liquid infects cells in a 96-pore plate, a sample to be tested is diluted by virus culture solution (DMEM +2% FBS +1% P/S), the infection is simultaneously dosed for determination, the sample to be tested is tested by samples diluted by 8 doses three times, each dose is provided with 2 parallel pores, when the lesion of a virus control group reaches No. 4+, the result is observed, the half inhibition concentration and the selection index of the sample to be tested on the virus are recorded and calculated by a Reed-Muench method (SI = EC)₅₀/ CC₅₀). To evaluate the cytotoxic toxicity of the reference compound (ribavirin) and KH-0016, a cytotoxicity control experiment was set up, as above, but no virus was added.

3.2 Experimental results:

TABLE 5 anti-coronavirus experimental results (EC 50)

Compound (I)	EC50
		Ribavirin (mu M)	18.61
KH-0016（mg/ml）	0.36

TABLE 6 cytotoxic assay results of test substances against Huh7 cells (CC50)

Compound (I)	CC50
		Ribavirin (mu M)	>100
KH-0016（mg/ml）	>2.27

The result shows that KH-0016 has stronger anti-coronavirus activity, and the selection index SI is more than 6.3.

Through repeated experiments, the products prepared by other embodiments of the invention can obtain the same or similar beneficial effects as the experimental results, and have obvious effects of resisting respiratory syncytial virus, influenza virus and coronavirus.

4. Conclusion

The experimental results show that the functional plant extract can effectively inhibit the proliferation of respiratory viruses including but not limited to respiratory syncytial virus, influenza virus and coronavirus in cells, and can be used as a medicinal and edible plant extract.

Claims

1. A functional plant extract with respiratory virus resistance is characterized in that the functional plant extract is prepared from the following raw materials in parts by weight: 1-30 parts of honeysuckle, 1-30 parts of chrysanthemum, 1-30 parts of dandelion, 1-30 parts of astragalus, 1-30 parts of schizonepeta, 1-30 parts of liquorice, 1-30 parts of pogostemon cablin, 1-30 parts of fried semen coicis, 1-30 parts of Chinese date, 1-30 parts of mint and 0.001-0.1 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus, the liquorice, the fried semen coicis and the Chinese date together, adding 10-250 times of purified water for soaking for 30-60 minutes, then boiling for 10-60 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuing to decoct for 10-60 minutes, adding the sucralose after clarifying and cooling to room temperature, filtering, subpackaging and sterilizing to obtain the traditional Chinese medicine.

2. The functional plant extract with the respiratory tract virus resisting function as claimed in claim 1, is characterized by being prepared from the following raw materials in parts by weight: 10 parts of honeysuckle, 10 parts of chrysanthemum, 10 parts of dandelion, 10 parts of astragalus membranaceus, 10 parts of schizonepeta, 10 parts of liquorice, 10 parts of pogostemon cablin, 10 parts of fried semen coicis, 10 parts of Chinese date, 10 parts of mint and 0.02 part of sucralose, wherein the honeysuckle, the chrysanthemum, the dandelion, the astragalus membranaceus, the liquorice, the fried semen coicis and the Chinese date are mixed together, 10 times of purified water is added for soaking for 30 minutes, then the mixture is boiled for 30 minutes, the schizonepeta, the pogostemon cablin and the mint are added, the decoction is continued for 30 minutes, the sucralose is added after the mixture is clarified and cooled to the.

3. The functional plant extract with the respiratory tract virus resisting function as claimed in claim 1, is characterized by being prepared from the following raw materials in parts by weight: 20 parts of honeysuckle, 20 parts of chrysanthemum, 20 parts of dandelion, 20 parts of astragalus membranaceus, 20 parts of schizonepeta, 20 parts of liquorice, 20 parts of pogostemon cablin, 20 parts of fried semen coicis, 20 parts of Chinese date, 20 parts of mint and 0.05 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus membranaceus, the liquorice, the fried semen coicis and the Chinese date together, adding 50 times of purified water, soaking for 30 minutes, then boiling for 30 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuously decocting for 30 minutes, adding the sucralose after clarifying and cooling to room temperature, filtering, subpackaging and sterilizing to obtain the.

4. The functional plant extract with the respiratory tract virus resisting function as claimed in claim 1, is characterized by being prepared from the following raw materials in parts by weight: 30 parts of honeysuckle, 30 parts of chrysanthemum, 30 parts of dandelion, 30 parts of astragalus, 30 parts of schizonepeta, 30 parts of liquorice, 30 parts of pogostemon cablin, 30 parts of fried semen coicis, 30 parts of Chinese date, 30 parts of mint and 0.1 part of sucralose, firstly mixing the honeysuckle, the chrysanthemum, the dandelion, the astragalus, the liquorice, the fried semen coicis and the Chinese date together, adding 250 times of purified water, soaking for 30 minutes, then boiling for 30 minutes, then adding the schizonepeta, the pogostemon cablin and the mint, continuing to decoct for 30 minutes, adding the sucralose after clarifying and cooling to room temperature, filtering, subpackaging and sterilizing to obtain the traditional Chinese medicine.