CN116474032A - Traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof - Google Patents

Traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof Download PDF

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Publication number
CN116474032A
CN116474032A CN202310471356.2A CN202310471356A CN116474032A CN 116474032 A CN116474032 A CN 116474032A CN 202310471356 A CN202310471356 A CN 202310471356A CN 116474032 A CN116474032 A CN 116474032A
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parts
essential oil
oil composition
chinese medicine
traditional chinese
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Inventor
董文化
王昊
梅文莉
戴好富
李薇
蔡彩虹
陈惠琴
曾军
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Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
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Institute of Tropical Bioscience and Biotechnology Chinese Academy of Tropical Agricultural Sciences
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    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/83—Thymelaeaceae (Mezereum family), e.g. leatherwood or false ohelo
    • A61K36/835—Aquilaria
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    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
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    • A—HUMAN NECESSITIES
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Abstract

The invention provides a traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof. A traditional Chinese medicine essential oil composition comprises the following raw materials in parts by weight: 10-20 parts of agilawood, 10-20 parts of Hainan phyllanthus urinaria, 10-15 parts of white mulberry root-bark, 10-15 parts of mugwort leaf, 10-15 parts of oriental orange, 5-10 parts of baical skullcap root, 5-10 parts of perilla, 5-10 parts of calamus, 3-5 parts of wrinkled gianthyssop herb, 3-5 parts of oldenlandia diffusa and 3-5 parts of rhizoma atractylodis. The Chinese medicinal essential oil composition not only has the effect of resisting influenza viruses, but also has obvious antibacterial and anti-inflammatory activities, and can be well applied to preparing medicaments for preventing and treating the influenza viruses.

Description

Traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof
Technical Field
The invention relates to the field of traditional Chinese medicine essential oil, in particular to a traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof.
Background
Traditional Chinese medicine considers that epidemic disease prevention is greater than treatment. For a long time, there are traditional fumigating Chinese medicines for eliminating cataract, preventing epidemic and avoiding infection. Research shows that the traditional Chinese medicine is a key active substance for playing the efficacy of the traditional Chinese medicine, and the traditional Chinese medicine stimulates olfactory conduction through the volatilization of the medicine property, can improve the respiratory system function, inhibit influenza virus, play a role in preventing influenza, promote the immunity of human body and is particularly important for preventing viruses in the air. Some of the Chinese medicinal essential oils have a certain antiviral effect, but often have low antiviral activity, but often need high dosage, but have a certain cytotoxic activity, and are difficult to meet the requirements of obvious effect and safety.
Disclosure of Invention
In view of the above, the invention provides a traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof, and the composition has obvious antiviral activity.
The technical scheme of the invention is realized as follows:
a traditional Chinese medicine essential oil composition comprises the following raw materials in parts by weight: 10-20 parts of agilawood, 10-20 parts of Hainan phyllanthus urinaria, 10-15 parts of white mulberry root-bark, 10-15 parts of mugwort leaf, 10-15 parts of oriental orange, 5-10 parts of baical skullcap root, 5-10 parts of perilla, 5-10 parts of calamus, 3-5 parts of wrinkled gianthyssop herb, 3-5 parts of oldenlandia diffusa and 3-5 parts of rhizoma atractylodis.
The further technical scheme is that the traditional Chinese medicine essential oil composition comprises the following raw materials in parts by weight: 15 parts of agilawood, 15 parts of Hainan phyllanthus urinaria, 13 parts of white mulberry root-bark, 13 parts of mugwort leaf, 13 parts of oriental orange, 8 parts of baical skullcap root, 8 parts of perilla, 8 parts of calamus, 4 parts of wrinkled gianthyssop herb, 4 parts of oldenlandia diffusa and 4 parts of rhizoma atractylodis.
The invention provides a preparation method of a traditional Chinese medicine essential oil composition, which comprises the following steps: the preparation method comprises the steps of respectively taking Chinese medicinal materials of agilawood, folium hainanensis, cortex mori, folium artemisiae argyi, oriental orange, scutellaria baicalensis, perilla, calamus, wrinkled gianthyssop herb, oldenlandia diffusa and rhizoma atractylodis, extracting by flash to obtain absolute oil, and purifying the absolute oil by molecular distillation to obtain the target Chinese medicinal essential oil.
The further technical scheme is that the extraction solvent for flash extraction is 90% -95% v/v ethanol solution.
The further technical scheme is that the flash extraction temperature is 55-60 ℃, the rotation speed is 8000-9000r/min, the extraction time is 50-60 s, and the mass volume ratio kg/L of the feed liquid is 1: 20-30 parts.
The further technical scheme is that the molecular distillation extraction process comprises the following steps: the feeding rate is 3-5 mL/min, the distillation temperature is 80-90 ℃, the distillation pressure is 80-100 mTorr, and the film scraping speed is 80-100 r/min. The traditional Chinese medicine essential oil composition is applied to preparation of antibacterial medicines.
The traditional Chinese medicine essential oil composition is applied to preparation of anti-inflammatory drugs.
The traditional Chinese medicine essential oil composition is applied to preparation of anti-influenza virus medicines, and is preferably used for preparing anti-H1N 1 influenza virus medicines.
The traditional Chinese medicine essential oil composition is applied to the preparation of fumigates, stickers, sachets or gels.
Compared with the prior art, the invention has the beneficial effects that:
(1) The essential oil composition is scientifically compounded and has synergistic antiviral effect on influenza viruses. The invention combines the traditional Chinese medicine essential oil in a specific proportion, thereby effectively improving the anti-influenza virus effect. The Chinese medicinal essential oil composition has antiviral effect, has antibacterial and anti-inflammatory activities related to pneumonia and enteritis, has high biological safety, and can be well applied to preparing medicaments for preventing and treating influenza viruses.
(2) The essential oil composition can be also applied to fragrance fumigation, or made into slow-release gel, or processed into a fragrance paste or a fragrance bag, can be pasted on clothes, masks, portable articles and the like, can aromatic and prevent dirt, prevent influenza, relieve drowsiness, inhibit bacteria and remove odor, and pleasure, and is a simple, convenient and safe 'green' prevention method.
Detailed Description
In order to better understand the technical content of the present invention, the following provides specific examples to further illustrate the present invention.
The experimental methods used in the embodiment of the invention are conventional methods unless otherwise specified.
Materials, reagents, and the like used in the examples of the present invention are commercially available unless otherwise specified.
Example 1
A Chinese medicinal composition essential oil comprises the following raw materials in parts by weight: 15 parts of agilawood, 15 parts of Hainan phyllanthus urinaria, 13 parts of white mulberry root-bark, 13 parts of mugwort leaf, 13 parts of oriental orange, 8 parts of baical skullcap root, 8 parts of perilla, 8 parts of calamus, 4 parts of wrinkled gianthyssop herb, 4 parts of oldenlandia diffusa and 4 parts of rhizoma atractylodis.
The preparation method of the essential oil of the traditional Chinese medicine composition comprises the following steps:
(1) Respectively weighing lignum Aquilariae Resinatum, folium Callicarpae Formosanae, cortex Mori, folium Artemisiae Argyi, oriental orange, scutellariae radix, perillae herba, rhizoma Acori Calami, herba Agastaches, herba Hedyotidis Diffusae, and rhizoma Atractylodis, and pulverizing.
(2) The crushed traditional Chinese medicine materials in the step (1) are firstly subjected to flash extraction, the extraction temperature is 60 ℃, the extraction time is 50s, the extraction solvent is 90% v/v ethanol solution, the rotating speed is 8000r/min, and the mass-volume ratio kg/L of the feed liquid is 1:20, collecting the extract, concentrating under reduced pressure, and recovering ethanol to obtain purified oil.
(3) Extracting the purified oil in the step (2) through molecular distillation, and carrying out a molecular distillation purification process: the feeding rate is 3mL/min, the distillation temperature is 80 ℃, the distillation pressure is 80mTorr, the film scraping speed is 80r/min, and the light component is collected to obtain the target traditional Chinese medicine essential oil.
Example 2
A Chinese medicinal composition essential oil comprises the following raw materials in parts by weight: 10 parts of agilawood, 20 parts of Hainan phyllanthus urinaria, 10 parts of white mulberry root-bark, 15 parts of mugwort leaf, 10 parts of oriental orange, 10 parts of baical skullcap root, 5 parts of perilla, 10 parts of calamus, 3 parts of wrinkled gianthyssop herb, 5 parts of oldenlandia diffusa and 3 parts of rhizoma atractylodis. The preparation method of the essential oil of the traditional Chinese medicine composition is the same as that of the embodiment 1.
Example 3
A Chinese medicinal composition essential oil comprises the following raw materials in parts by weight: 20 parts of agilawood, 10 parts of Hainan phyllanthus urinaria, 15 parts of white mulberry root-bark, 10 parts of folium artemisiae argyi, 15 parts of oriental orange, 5 parts of radix scutellariae, 10 parts of perilla, 5 parts of calamus, 5 parts of wrinkled gianthyssop, 3 parts of oldenlandia diffusa and 5 parts of rhizoma atractylodis. The preparation method of the essential oil of the traditional Chinese medicine composition is the same as that of the embodiment 1.
Example 4
A Chinese medicinal composition essential oil is consistent with example 1, and comprises the following raw materials in parts by weight: 15 parts of agilawood, 15 parts of Hainan phyllanthus urinaria, 13 parts of white mulberry root-bark, 13 parts of mugwort leaf, 13 parts of oriental orange, 8 parts of baical skullcap root, 8 parts of perilla, 8 parts of calamus, 4 parts of wrinkled gianthyssop herb, 4 parts of oldenlandia diffusa and 4 parts of rhizoma atractylodis.
The preparation method of the essential oil of the traditional Chinese medicine composition comprises the following steps:
(1) Respectively weighing lignum Aquilariae Resinatum, folium Callicarpae Formosanae, cortex Mori, folium Artemisiae Argyi, oriental orange, scutellariae radix, perillae herba, rhizoma Acori Calami, herba Agastaches, herba Hedyotidis Diffusae, and rhizoma Atractylodis, and pulverizing.
(2) The crushed traditional Chinese medicine materials in the step (1) are firstly subjected to flash extraction, wherein the extraction temperature is 58 ℃, the extraction time is 55s, the extraction solvent is 93% v/v ethanol solution, and the rotation speed is 8500r/min, and the mass-volume ratio kg/L of the feed liquid is 1:25, concentrating under reduced pressure to recover ethanol to obtain absolute oil.
(3) Extracting the extracting solution in the step (2) through molecular distillation, and carrying out a molecular distillation purification process: the feeding rate is 4mL/min, the distillation temperature is 85 ℃, the distillation pressure is 90mTorr, the film scraping speed is 90r/min, and the light component is collected to obtain the target traditional Chinese medicine essential oil.
Example 5
A Chinese medicinal composition essential oil is consistent with example 1, and comprises the following raw materials in parts by weight: 15 parts of agilawood, 15 parts of Hainan phyllanthus urinaria, 13 parts of white mulberry root-bark, 13 parts of mugwort leaf, 13 parts of oriental orange, 8 parts of baical skullcap root, 8 parts of perilla, 8 parts of calamus, 4 parts of wrinkled gianthyssop herb, 4 parts of oldenlandia diffusa and 4 parts of rhizoma atractylodis.
The preparation method of the essential oil of the traditional Chinese medicine composition comprises the following steps:
(1) Respectively weighing lignum Aquilariae Resinatum, folium Callicarpae Formosanae, cortex Mori, folium Artemisiae Argyi, oriental orange, scutellariae radix, perillae herba, rhizoma Acori Calami, herba Agastaches, herba Hedyotidis Diffusae, and rhizoma Atractylodis, and pulverizing.
(2) The crushed traditional Chinese medicine materials in the step (1) are firstly subjected to flash extraction, the extraction temperature is 60 ℃, the extraction time is 60s, the extraction solvent is 95% v/v ethanol solution, the rotation speed is 9000r/min, and the mass-volume ratio kg/L of the feed liquid is 1:30, concentrating under reduced pressure to recover ethanol to obtain absolute oil.
(3) Extracting the extracting solution in the step (2) through molecular distillation, and carrying out a molecular distillation purification process: the feeding rate is 5mL/min, the distillation temperature is 90 ℃, the distillation pressure is 100mTorr, the film scraping speed is 100r/min, and the light component is collected to obtain the target traditional Chinese medicine essential oil.
Comparative example 1
A Chinese medicinal composition essential oil comprises the following raw materials in parts by weight: 10 parts of agilawood, 10 parts of Hainan phyllanthus urinaria, 10 parts of white mulberry root-bark, 10 parts of mugwort leaf, 10 parts of oriental orange, 10 parts of baical skullcap root, 10 parts of perilla, 10 parts of calamus, 10 parts of wrinkled gianthyssop herb, 10 parts of oldenlandia diffusa and 10 parts of rhizoma atractylodis. The preparation method of the essential oil of the traditional Chinese medicine composition is the same as that of the embodiment 1.
The essential oils of the traditional Chinese medicine compositions prepared in the above examples 1-5 and comparative example 1 were subjected to antibacterial, anti-inflammatory and antiviral tests, and the results are as follows:
1 antibacterial test
Adopting a filter paper agar diffusion method to test bacteria: staphylococcus aureus, drug to be tested: real worldThe essential oil of the Chinese medicinal composition of examples 1-5, kanamycin sulfate, served as a positive control. Preparing chloroform methanol solution with concentration of 10mg/mL from Chinese medicinal composition essential oil and kanamycin sulfate, respectively dripping 50 μl of medicinal solution and kanamycin sulfate onto sterilized filter paper sheets (diameter of 6 mm), volatilizing solvent, and attaching the spotted filter paper sheets to 100 μl (bacterial liquid concentration about 10) of coated test bacterial suspension 6 CFU/mL), placing for 30min, placing into an incubator, culturing at 36 deg.C without light, observing and measuring the diameter of the drug inhibition zone after 24 h. Each sample was repeated 3 times.
TABLE 1 antibacterial test results
The results show that examples 1-5 and comparative example 1 all have a certain antibacterial effect. Wherein the essential oil compositions of examples 1-5 have a significant antimicrobial effect. Compared with the embodiment 1, the antibacterial effect of the comparative example 1 is reduced, and further shows that the antibacterial effect is effectively improved by adopting the traditional Chinese medicine essential oil combined in a specific proportion.
2 anti-inflammatory test
Inhibiting Lipopolysaccharide (LPS) -induced activity of mouse mononuclear macrophage RAW264.7 to generate NO establishes LPS-induced mouse mononuclear macrophage RAW264.7 model, and adopts Griess method to test anti-inflammatory activity of essential oil of the Chinese medicinal composition.
Cytotoxic Activity test: the cytotoxic activity of the essential oil of the traditional Chinese medicine composition on the mouse mononuclear macrophage RAW264.7 is tested by adopting an MTT method. RAW264.7 cells were cultured with DMEM medium containing 10% FBS at 37deg.C and 5% CO 2 Culturing in an incubator conventionally. Cell count 1X 10 5 After inoculating the sample/mL to a 96-well plate for 24 hours, adding 100 mu L of a drug solution to be tested (100 mu mol/L), placing the mixture at 37 ℃ and 5% CO 2 After the cells were attached to the wall in the cell incubator for 72 hours, 15. Mu.L of MTT solution (5 mg/mL) was added to each well of cells, the supernatant was aspirated after the reaction at 37℃for 4 hours, 100. Mu.L of LDMSO was added to each well to be sufficiently dissolved, and the absorbance (A) value of each well was measured at 490nm wavelength by an ELISA reader, and each set of experiments was performed in parallel for 3 times. The cell growth inhibition was calculated according to the formula. Inhibition ratio =1-A 1 /A 0 ；A 0 Absorbance of the blank (100. Mu.L of culture solution), A 1 Is the absorbance of the drug group (100. Mu.L drug solution).
The results show that the essential oils of the Chinese medicinal compositions of examples 1-5 and comparative example 1 do not have cytotoxic activity at a concentration of 100. Mu. Mol/L.
The medicine to be tested: essential oils of the Chinese medicinal compositions of examples 1 to 5 and comparative example 1. Positive control indomethacin and quercetin were compared. RAW264.7 cells were cultured with DMEM medium containing 10% FBS at 37deg.C and 5% CO 2 Culturing in an incubator conventionally. Cell count 1X 10 5 Inoculating the mixture to 96-well plate, culturing for 24 hr, adding 50 μl of the medicinal solution (with medicinal concentration of 100, 50, 25, 12.5, 6.25 μmol/L), culturing for 1 hr, adding 50 μLLPS, standing at 37deg.C and 5% CO 2 After 24h of adherence in a cell incubator, 100. Mu.L of Griess reagent was added to each well in a new 96-well plate, and after mixing, the A of each well was measured at 540nm wavelength by an ELISA reader, and each experiment was performed 3 times in parallel. And calculating the NO inhibition rate according to a formula. Finally, the data were processed using GraphPadPrism software to determine the semi-Inhibitory Concentration (IC) of the compound on LPS-induced RAW264.7 production of NO 50 )。
Inhibition ratio = (A2-A1)/(A2-A0)
A0 is absorbance of a blank group (50. Mu.L of culture solution + 50. Mu.L of culture solution),
a1 is the absorbance of the drug group (50. Mu.L drug+50. Mu.L lipopolysaccharide),
a2 shows the absorbance test result of LPS-induced group (50. Mu.L of culture medium+50. Mu.L of lipopolysaccharide), and Table 2 shows the anti-inflammatory test result
The results show that the traditional Chinese medicine essential oil compositions in examples 1-5 have obvious anti-inflammatory activity. Inhibiting LPS-induced mouse megaphaga-niveaThe cell RAW264.7 has stronger activity of generating NO and IC 50 The value is 17.64-55.45 mu mol/L.
3 anti-influenza Virus test
50 SPF grade ICR mice are selected, and the male and female mice are half, 9-12 weeks old and 13-15g in weight. After 3d of adaptive feeding, the feed was used for experiments. The groups were randomized, with 10 groups each, including a healthy control group, a model control group, and three low, medium, and high doses of essential oil of the herbal composition (example 1). In addition to the healthy control group, mice were lightly anesthetized with diethyl ether and infected with 15 drops of LD50 influenza virus liquid (H1N 1/PR8 strain) at a drop rate of 35 μl/mouse. The administration was started on day 1 of infection, and the low, medium and high dose groups were dosed at 2ml/kg, 4ml/kg, 8ml/kg, respectively, for 4 consecutive days, once daily. The model control group and the healthy control group were given by nasal administration of pure water drops under the same conditions. Dissecting after weighing on the 5 th day, taking the lung, weighing the lung, and calculating the lung index and the lung index inhibition rate. The calculation formula is as follows:
lung index (%) = (wet lung weight/body weight) ×100%;
lung index inhibition (%) = (model control lung weight-dosing lung weight)/(model control lung weight-healthy control lung weight) ×100%.
The results are shown in Table 3
TABLE 3 anti-influenza Virus test results
The results show that the lung index of the medium and high concentration dosage groups of the essential oil composition prepared by the invention is obviously reduced, and compared with a control group of a model group, the essential oil composition has obvious difference (P is less than 0.01), which indicates that the essential oil composition prepared by the invention has a therapeutic effect on a pneumonia model of mice infected with influenza virus.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, alternatives, and improvements that fall within the spirit and scope of the invention.

Claims (10)

1. The traditional Chinese medicine essential oil composition is characterized by comprising the following raw materials in parts by weight: 10-20 parts of agilawood, 10-20 parts of Hainan phyllanthus urinaria, 10-15 parts of white mulberry root-bark, 10-15 parts of mugwort leaf, 10-15 parts of oriental orange, 5-10 parts of baical skullcap root, 5-10 parts of perilla, 5-10 parts of calamus, 3-5 parts of wrinkled gianthyssop herb, 3-5 parts of oldenlandia diffusa and 3-5 parts of rhizoma atractylodis.
2. The traditional Chinese medicine essential oil composition according to claim 1, which is characterized by comprising the following raw materials in parts by weight: 15 parts of agilawood, 15 parts of Hainan phyllanthus urinaria, 13 parts of white mulberry root-bark, 13 parts of mugwort leaf, 13 parts of oriental orange, 8 parts of baical skullcap root, 8 parts of perilla, 8 parts of calamus, 4 parts of wrinkled gianthyssop herb, 4 parts of oldenlandia diffusa and 4 parts of rhizoma atractylodis.
3. The essential oil composition of claim 1, wherein the preparation method of the essential oil composition comprises the following steps: the preparation method comprises the steps of respectively taking Chinese medicinal materials of agilawood, folium hainanensis, cortex mori, folium artemisiae argyi, oriental orange, radix scutellariae, perilla, calamus, wrinkled gianthyssop herb, oldenlandia diffusa and rhizoma atractylodis, extracting by flash to obtain absolute oil, and purifying the absolute oil by molecular distillation to obtain the target Chinese medicinal essential oil.
4. The essential oil composition of claim 3, wherein the flash extraction solvent is 90% -95% v/v ethanol solution.
5. The essential oil composition of claim 3, wherein the flash extraction temperature is 55-60 ℃, the rotation speed is 8000-9000r/min, the extraction time is 50-60 s, and the feed-liquid ratio is 1: 20-30 parts.
6. The essential oil composition of claim 3, wherein the molecular distillation extraction process: the flow rate is 3-5 mL/min, the distillation temperature is 80-90 ℃, the distillation pressure is 80-100 mTorr, and the film scraping speed is 80-100 r/min.
7. The use of a traditional Chinese medicine essential oil composition according to any one of claims 1-6 in the preparation of anti-influenza virus drugs.
8. The use of the essential oil composition of claim 7 in the preparation of anti-H1N 1 influenza virus drugs.
9. The use of a Chinese medicinal essential oil composition according to any of claims 1-6 in the preparation of an antibacterial medicament.
10. The use of a traditional Chinese medicine essential oil composition according to any of claims 1-6 in the preparation of anti-inflammatory drugs.
CN202310471356.2A 2023-04-27 2023-04-27 Traditional Chinese medicine essential oil composition for preventing and treating influenza virus and application thereof Pending CN116474032A (en)

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