CN114246925A

CN114246925A - Traditional Chinese medicine essential oil and application thereof

Info

Publication number: CN114246925A
Application number: CN202011002501.5A
Authority: CN
Inventors: 詹华强; 梁嘉荣; 邓伟麟; 段然; 董婷霞; 肖健; 徐启乐
Original assignee: Hong Kong University of Science and Technology HKUST
Current assignee: Hong Kong University of Science and Technology HKUST
Priority date: 2020-09-22
Filing date: 2020-09-22
Publication date: 2022-03-29

Abstract

The invention provides traditional Chinese medicine essential oil, wherein the traditional Chinese medicine essential oil comprises one or more of the following traditional Chinese medicine extracts: clove, cinnamon, curcuma zedoary, perilla leaf, magnolia flower, citronella, common cnidium fruit, eupatorium, liquorice, honeysuckle, forsythia, dandelion and radix bupleuri, and application of the traditional Chinese medicine essential oil in preparing a preparation for preventing coronavirus infection and/or bacterial infection. The invention provides a method for inhibiting coronavirus and/or bacteria, which comprises the step of contacting the traditional Chinese medicine essential oil with the coronavirus and/or the bacteria. In addition, the invention also provides a daily necessity containing the traditional Chinese medicine essential oil.

Description

Traditional Chinese medicine essential oil and application thereof

Technical Field

The invention relates to the technical field of medicines, in particular to traditional Chinese medicine essential oil, application thereof in preparing a preparation for preventing coronavirus infection and/or bacterial infection, a method for inhibiting coronavirus and/or bacteria, and daily necessities containing the traditional Chinese medicine essential oil.

Background

Coronaviruses are a large family of viruses known to cause the common cold and more serious diseases such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). The novel coronavirus is a new strain of coronavirus that has not been previously discovered in humans. Since 12 months in 2019, monitoring of influenza and related diseases is continuously carried out in Wuhan City in Hubei province, and multiple viral pneumonia cases are found and are diagnosed as viral pneumonia/lung infection. The international committee for virus classification states that this new pathogen will be named severe acute respiratory syndrome coronavirus No. 2 (severe acute respiratory syndrome coronavirus 2) or SARS-CoV-2, which is simply referred to as "new coronavirus" in the present invention.

Since the outbreak of the new coronavirus (SARS-CoV-2), a global pandemic was caused. Nearly 1500 million people are infected in the world by 7 middle ten days of 2020, and more than 60 people lose life, resulting in inestimable loss. After a person is infected with a new coronavirus (SARS-CoV-2), the common signs of the person include respiratory symptoms, fever, cough, shortness of breath, dyspnea and the like. In more severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, renal failure, and even death. At present, no specific treatment method exists for diseases caused by the new coronavirus (SARS-CoV-2), no specific antiviral medicine for the new coronavirus (SARS-CoV-2) exists, and no breakthrough progress exists in vaccine development.

Therefore, the discovery of a drug candidate for inhibiting the novel coronavirus (SARS-CoV-2) is imminent.

Disclosure of Invention

In view of the above, the present invention provides a traditional Chinese medicine essential oil and its use in preparation of a preparation for preventing coronavirus infection and/or bacterial infection, a method for inhibiting coronavirus and/or bacteria, and a daily necessity containing the traditional Chinese medicine essential oil.

According to a first aspect of the invention, the invention provides a traditional Chinese medicine essential oil, wherein the traditional Chinese medicine essential oil comprises one or more of the following traditional Chinese medicine extracts: clove, cinnamon, curcuma zedoary, perilla leaf, magnolia flower, citronella, fructus cnidii, eupatorium, liquorice, honeysuckle, fructus forsythiae, dandelion and radix bupleuri.

According to a second aspect of the present invention there is provided a method of inhibiting coronavirus and/or bacteria, said method comprising contacting an essential oil of a Chinese medicinal material according to the first aspect of the present invention with coronavirus and/or bacteria.

According to a third aspect of the present invention, there is provided the use of an essential oil of a Chinese medicinal material according to the first aspect of the present invention in the manufacture of a formulation for the prevention of coronavirus infection and/or bacterial infection.

According to a fourth aspect of the invention, the invention provides a daily necessity, which comprises the traditional Chinese medicine essential oil of the first aspect of the invention.

The invention has the beneficial effects that:

the invention uses the extracts of a plurality of Chinese medicinal materials to prepare the Chinese medicinal essential oil with the effects of resisting bacteria, viruses and the like, thereby being used for inhibiting coronavirus and/or bacteria and preventing and/or treating coronavirus infection and/or bacterial infection. The Chinese medicinal essential oil provided by the invention not only contains Chinese medicinal extracts, for example, through supercritical treatmentBoundary CO₂The extract of Chinese medicinal materials obtained by extraction or steam distillation, and base oil commonly used in essential oil preparation, can not only give fragrant smell, but also have antibacterial and antiviral effects.

The traditional Chinese medicine essential oil is also applied to daily necessities such as masks, disinfectant, aromatherapy, spray, hand sanitizer or skin care products, has antibacterial and antiviral effects and aromatic taste, can make people happy, helps people prevent virus invasion, enhances the upper respiratory tract defense capacity, and simultaneously can keep physical and mental health of people.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other embodiments can be obtained according to the drawings without inventive labor.

FIG. 1 is a graphical representation of the results of the anti-neocoronaviruses (SARS-CoV-2) effect of low concentrations of essential oils (0.1% by volume) according to some embodiments of the invention;

FIG. 2 is a graphical representation of the results of the anti-neocoronaviruse (SARS-CoV-2) effect of high concentrations of essential oils (1% by volume) according to some embodiments of the invention;

FIG. 3 is a graphical representation of the results of antiviral effects of different concentrations of essential oils (0.003%, 0.01%, 0.1%, 0.3%, 1%, by volume percent) according to some embodiments of the present invention;

fig. 4 is a graph showing the results of antibacterial activities of five essential oils of clove, zedoary, cinnamon, bay leaf and citronella of example 3, wherein SA: staphylococcus aureus bacteria; EC: e.coli; KP: klebsiella pneumoniae; PA: pseudomonas aeruginosa; BS: b, bacillus subtilis; ah: aeromonas hydrophila; et: edwardsiella tarda; va: vibrio alginolyticus; and Vh: vibrio harveyi.

Fig. 5 is a schematic view showing an open structure of a mask according to some embodiments of the present invention, in which 1 denotes a metal wire, 2 denotes a medicated wire, 3 denotes an outer layer, 4 denotes a middle layer, and 5 denotes an inner layer.

Detailed Description

The present invention will now be described more fully hereinafter with reference to the accompanying drawings, in which embodiments of the invention are shown. It is to be understood that the described embodiments are merely a subset of the present invention and not all embodiments. All other embodiments that can be derived by one of ordinary skill in the art from the embodiments disclosed herein are within the scope of the present invention.

The new coronavirus (SARS-CoV-2) has caused an infectious disease which is prevalent in the world, however, there is no specific treatment for the disease caused by the new coronavirus and no antiviral drug specific to the new coronavirus at present. Therefore, the discovery of candidate drugs capable of inhibiting coronavirus or further a novel coronavirus (SARS-CoV-2) is imminent.

The traditional Chinese medicine is a substance for preventing and treating diseases and having rehabilitation and health care functions under the guidance of the theory of traditional Chinese medicine, and is an important substance for preventing and treating diseases and resisting epidemic historically. The habit of inserting folium artemisiae argyi and wearing a fragrant bag at noon has been existed since ancient times, and the fragrant bag has the functions of resuscitation, dirt avoidance and epidemic avoidance. The Chinese herbs have several properties and actions to act on the disease condition, and many Chinese herbs have antibacterial and antiviral effects.

The essential oil is extracted from leaves, flowers, seeds, fruits, roots, barks, resins, hearts and other parts of plants by steam distillation, cold pressing, fat absorption and solvent extraction, and has high-concentration fragrance and volatility. The pure natural plant essential oil has fragrant smell, and after entering the brain, the natural aromatic essential oil can stimulate the anterior lobe of the brain to secrete two hormones, namely endorphin and enkephalin, so that the spirit is in a comfortable state, and the essential oil also has the effects of sterilizing, resisting inflammation, healing, deodorizing, tranquilizing, expelling parasites, softening and moistening the fine skin and the like. Although the chemical compositions of various essential oils are different, due to similar synthetic routes, a plurality of chemical components can be found in various plant essential oils, and the essential oils have the efficacies of sterilization, anti-inflammation and the like and also contain the chemical components with biological activity.

Based on the above, the invention uses the extracts of a plurality of Chinese medicinal materials to prepare the Chinese medicinal essential oil with antibacterial and antiviral effects, thereby being used for inhibiting coronavirus and/or bacteria and preventing and/or treating coronavirus infection and/or bacterial infection.

Thus, according to a first aspect of the present invention, there is provided a Chinese medicinal essential oil comprising one or more of the following extracts of Chinese herbs: clove, cinnamon, curcuma zedoary, perilla leaf, magnolia flower, citronella, fructus cnidii, eupatorium, liquorice, honeysuckle, fructus forsythiae, dandelion and radix bupleuri.

Flos Caryophylli, also known as Dingzi Xiang, Zhijixiang and flos Caryophylli. The bud of clove is a common traditional Chinese medicine. Modern researches show that the clove contains 15-20% of volatile oil, the main components in the oil comprise eugenol, beta-caryophyllene, acetyl eugenol and the like, and the oil also contains components of eugenol, acetyl eugenol, isoquercitin, kaempferol, quercetin, caryophyllene oxide, oleanolic acid, alpha-menthene and the like. Flos Caryophylli has stomach invigorating effect, and the leachate has obvious function of stimulating gastric secretion, and can relieve abdominal distention, nausea, emesis, etc. In addition, it has inhibitory effect on pathogenic fungi, coccus, streptococcus, and Bacillus such as pneumonia, dysentery, large intestine, typhoid fever, and influenza virus.

Cinnamon, also known as cinnamon, Cinnamomum cassia, is a dry bark of Cinnamomum cassia Presl, a plant of lauraceae. Cinnamon has a mild stimulation effect on intestines and stomach, can relieve gastrointestinal smooth muscle spasm, and has a strong anti-ulcer effect; can antagonize platelet aggregation, improve cardiovascular system, and regulate immunity; has tranquilizing, analgesic, antipyretic, and anticonvulsive effects on central nervous system; can be used for preventing and protecting yang deficiency and yin deficiency; has certain anti-inflammatory effect; has strong antifungal effect.

Rhizoma Curcumae, also known as Wen zedoary, rhizoma Curcumae, rhizoma Et radix Notopterygii, rhizoma Curcumae, rhizoma Zingiberis recens, and rhizoma Curcumae, mainly contains volatile oil, flavonoids, and terpenes. The Curcumae rhizoma mainly comprises curcumenone, curcumenol, Borneolum, radix et rhizoma Rhei cage for animals geranone, Camphora, curcumene, turmerone, curdione, cineole, daucosterol, beta-sitosterol, and palmitic acid. Curcumae rhizoma has effects of resisting tumor, resisting bacteria, increasing leukocyte, promoting blood circulation, and removing blood stasis; for the low-concentration tension and high-concentration relaxation of gastrointestinal smooth muscle; has liver protecting effect; improving acute renal failure; inhibiting platelet aggregation and antithrombotic formation; has anti-inflammatory effect.

Perilla leaf, also known as folium Perillae, thallus Porphyrae, and Su, contains perillaldehyde, perillaketone, perilla ketone, menthol, alpine baicalin, New Zealand vitexin II, and lukunin. Folium Perillae has tranquilizing and antipyretic effects; can inhibit the excitability membrane; can promote secretion of digestive juice and enhance gastrointestinal peristalsis; has the functions of relieving cough, eliminating phlegm and relieving asthma on the respiratory system and has the double functions of stopping bleeding and resisting coagulation on the blood system; increase blood glucose; regulating organism immunity, and resisting mutagenesis; has antimicrobial, broad-spectrum antibacterial, and antiinflammatory effects.

Flos Magnoliae, also called magnolia or magnolia liliiflora, contains volatile oil, the main component of the oil is 1, 8-cineole, etc., and the other components of beta-pinene, 1, 8-cineole, camphor, hederagenin, bergamotene, magnolia biondin A, etc., and has the functions of anesthesia, antianaphylaxis, anti-inflammation, blood pressure reduction, uterine excitation, anti-platelet aggregation, microorganism resistance, pain relief and microcirculation improvement.

The citronella, namely the lemongrass, the lemongrass and the lemongrass, the stem and leaf contain 0.4 to 0.8 percent of volatile oil, wherein the content of the citral serving as a main component is 75 to 85 percent, and a small amount of citronellal, geraniol and methyl heptenone are contained. The volatile oil of herba Cymbopogonis Citrari has antifungal effect (such as Streptomyces venezuelae). Has good effect of treating rheumatism, treating migraine, resisting infection, improving digestive function, deodorizing and expelling parasites. Resisting infection, astringing skin, and regulating greasy and unclean skin. It can give refreshing feeling and restore physical and mental balance (especially in early stage of disease recovery). Is essential oil with the most extensive application in aromatherapy and medical treatment. It can also be used as indoor aromatic.

Fructus Cnidii, also known as fructus Foeniculi, radix Dauci Sativae, flos Serpentis, SHEZHU, SHEXIAN, and fructus Cnidii. Contains volatile oil, which mainly comprises pinine, isovalerian borneol ester, parsley phenol methyl ether, dihydrocarveol, bergapten, cnidium lactone, isoanisic apigenin, etc. In addition, it contains components such as camphene, isoimperatorin, Columbia lactone, bornyl acetate, limonene, 3-sitosterol, etc. Fructus Cnidii has effect in resisting arrhythmia; inhibiting the heart; lowering blood pressure; a sex hormone-like effect; antifungal, insecticidal, trichomonal and spermicidal; dispel phlegm and relieve dyspnea; local anesthesia; anti-allergic reaction, anti-mutagenic effect; delaying aging; affecting the metabolism of plasma prostacyclin and inner cyclic nucleotides; inhibiting smooth muscle contraction; preventing and treating osteoporosis; has sedative and hypnotic effects.

Herba Eupatorii, herba Lycopi, herba Eupatorii, and herba Eupatorii chinensis containing volatile oil; the leaf contains coumarin, coumaric acid, and thymoquinol; the root contains vanillin. And other components such as aurantiol acetate, thymol methyl ether, p-cymene, taraxasterol acetate, and 3-sitosterol. Herba Eupatorii has expectorant, antiviral, and anticancer effects; has inhibitory effect on influenza virus; and can inhibit ovulation.

Licorice, also known as sweet licorice, licorice root, aged people and Mexican grass, mainly contains liquiritin, glycyrrhizic acid, glycyrrhizin, syringene oxide, glycyrrhetinic alcohol, 18 a-hydroxy glycyrrhetinic acid, isoglycyrrhetinic acid, glycyrrhizin, formononetin, neolicochalcone D, glabrene aglycone, isolicoflavonol, triterpenoid saponin, coumarin and other components. The licorice has the functions of adrenocortical hormone, anti-inflammation, anti-ulcer, anti-anaphylaxis, anti-cancer, anti-bacteria, anti-virus, promoting pancreatic secretion, inhibiting isolated intestine, regulating immunity, relieving cough, eliminating phlegm, resisting mutation, detoxifying, resisting oxidation, protecting the functions of ear and vestibule, promoting urination, protecting liver, preventing arteriosclerosis, resisting cerebral ischemia, preventing diabetic complications and the like.

Flos Lonicerae, also known as Lonicera Japonica flos, flos Lonicerae, diurea, flos Lonicerae, flos Celosiae Cristatae, and radix Ampelopsis Brevipedunculatae, and contains luteolin, inositol 1%, volatile oil, terpenes, and phenolic acid. The beverage contains chlorogenic acid, isochlorogenic acid, ginkgol, beta-sitosterol, stigmasterol-D-glucoside, etc. The flos Lonicerae has effects of resisting pathogenic microorganism, resisting virus, resisting inflammation, relieving fever, promoting phagocytic function of inflammatory cell, reducing blood lipid, exciting central nervous system, resisting fertility, preventing gastric ulcer, and exciting uterus. The decoction, water extract and purified liquid of honeysuckle flower have good killing and inhibiting effects on the streptococcus mutans causing dental caries by using a plate punching method, and the bacteriostatic effect is obviously enhanced along with the increase of the concentration.

Fructus forsythiae, also known as fructus forsythiae, HUANGHUATIAO, HUANGLIHUA, contains lignans, flavonoids, volatile components, phenethyls, ethyl cyclohexanol, triterpenes, and coumarins. Also comprises betulinic acid, phillyrin, arctiin, matairesinol, pinoresinol, phillyrin C, phillyrin D, etc. Fructus forsythiae has antimicrobial effect; inhibiting phosphodiesterase, lipoxygenase; stopping vomiting; anti-liver damage; anti-inflammatory; inhibiting elastase activity; reduce spontaneous hypertension, etc.

Herba Taraxaci, also called herba Taraxaci, herba Violae, herba Taraxaci, flos Hostae Plantagineae, flos Tagetis Erectae, herba Commelinae, flos Canarii albi, and herba Violae contains taraxasterol, choline, inulin, pectin, taraxasterol, taraxacin, and caffeic acid. Has effects in resisting pathogenic microorganism, resisting tumor, resisting gastric ulcer, promoting function of gallbladder, protecting liver, directly exciting heart of frog in vitro at low concentration, and inhibiting at high concentration. Can improve the tension of isolated duodenum and strengthen the contractility thereof, and is clinically considered to have the functions of strengthening stomach and laxation. The injection has strong bactericidal effect on staphylococcus aureus resistant strains and hemolytic streptococcus in a test tube, and also has a certain bactericidal effect on pneumococcus, meningococcus, diphtheria bacillus, pseudomonas aeruginosa, proteus, dysentery bacillus, typhoid bacillus and catacoccus.

Radix bupleuri, also known as bupleurum falcatum, bupleurum nanum, ground smoke, peruvian, mountain vegetable, rue grass and firewood, mainly contains valeric acid, linolenic acid, palmitic acid, stearic acid, kaempferol, kaempferitrin, quercetin, rutin, myrcene, borneol, saikosaponin II, myrtenol, alpha-spinasterol, deglucosyl saikosaponin alpha, saikosaponin and other components. And contains other components such as Chunfoushou alcohol. Pharmacological tests prove that the bupleurum has the effects of resisting inflammation, relieving fever, tranquilizing, relieving pain, relieving cough and resisting convulsion; can relieve liver injury and promote bile secretion; has effects in lowering blood pressure, reducing serum cholesterol, and promoting hemolysis; has antiulcer, antibacterial, antiviral, antitumor, blood sugar increasing, blood fat content reducing, and radiation injury resisting effects.

In a further embodiment, the extract of the chinese traditional medicine is extracted by supercritical CO₂Extracting with extraction method or steam distillation method.

Thus, in the present invention, "extract" means, for example, by using supercritical CO₂Extracting Chinese medicinal materials such as flos Caryophylli, cortex Cinnamomi, Curcumae rhizoma, folium Perillae, and flos Magnoliae by extraction or steam distillation to obtain extract containing low-polarity and volatile small molecule components. Of course, other methods generally used for extracting Chinese herbs known in the art may be used, and the present invention is not limited thereto.

Supercritical CO₂The extraction method comprises contacting supercritical fluid with substance to be separated under supercritical state to selectively extract polar components, boiling point components and molecular weight components sequentially. The method comprises pulverizing Chinese medicinal materials, adding into an extractor, adding a small amount of polar solvent (carrying agent) into the extractor, and allowing CO to flow at room temperature and low pressure (20-30 deg.C, 100-200bar)₂Supercritical fluid reaches subcritical fluid state, fluid is injected into the extractor from bottom to top to enter an extraction stage, after extraction is completed, extract liquor is decompressed and then enters a first separator and a second separator in sequence, and traditional Chinese medicine extract is collected at the bottom of the second separator.

Steam distillation is an extraction method in which a plant material containing volatile components is co-distilled with water to distill off the volatile components along with the steam, and the volatile components are separated by condensation. The crushed Chinese medicine is set inside a distiller with cooler connected to the upper part and steam connected to the lower part to maintain the steam pressure of 0.12-0.13 MPa. Vapor permeates the medicinal powder, volatile components in the medicinal powder are gasified due to the high temperature of the vapor, the vapor enters a cooler along with water vapor from a distillation tube, the vapor and the gasified volatile components are cooled in the cooler to form an oil-water mixture, the oil-water mixture is collected at the outlet of the cooler by an oil-water separator, after standing, oil and water are automatically separated, the upper layer is the traditional Chinese medicine extract, and the lower layer is water. Removing water from the lower layer with a separator to obtain the desired Chinese medicinal extract.

The Chinese medicinal essential oil may comprise extract of one Chinese medicinal material, or may comprise combination of extracts of multiple Chinese medicinal materials (such as two, three, four, five, six, seven, eight, nine, ten, eleven and twelve). It is understood that the herbal extracts can be combined in any ratio, and can be adjusted accordingly based on taste preferences of different people.

In one embodiment, the herbal essential oil comprises a combination of herbal extracts of: zedoary extract + cinnamon extract, bupleurum extract + cinnamon extract, licorice extract + clove extract, or eupatorium extract + bupleurum extract. Further, the zedoary extract + cinnamon extract, the bupleurum extract + cinnamon extract, the glycyrrhiza extract + clove extract or the eupatorium extract + bupleurum extract are combined in the ratio of (1-5): 5-1, for example, in the ratio of 1:5, 1:3, 1:2, 2:3, 1:1, 2:1, 3:1, 4:1, 5:1, preferably in the ratio of 1: 1.

As is well known to those skilled in the art, pure essential oils are very irritating, can cause damage when rubbed directly on the skin, and are very harmful, so the essential oil must be diluted before use on the skin. The base oil is a non-volatile oil extracted from seeds, flowers, rhizomes or fruits of plants, can lubricate the skin, can be directly used for skin massage, and is usually used for diluting simple essential oil (pure essential oil). The base oil can reduce irritation to skin and respiratory mucosa, slow down volatilization of essential oil, and prolong service life.

Thus, in one embodiment, the essential oil of Chinese traditional medicine further comprises one, two or three of sweet almond oil, rose seed oil or grape seed oil as a base oil. Preferably, the traditional Chinese medicine essential oil comprises grape seed oil. The grape seed oil has clear color and light taste, and can be suitable for different skins. Of course, other base oils known in the art may also be used.

In one embodiment, the concentration of each of the extracts in the herbal essential oils is 0.003% to 10% by volume, e.g., 0.005%, 0.008%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%, or any number between any two of these ranges.

In one embodiment, the concentration of each of the extracts in the herbal essential oil is 0.1% to 10%, for example 0.1% to 1%, by volume percentage.

In the present invention, the method may be a method for non-therapeutic purposes, for inhibiting, for example, coronavirus and/or bacteria in the ambient air, on the surface of an object, etc.; the method may also be a method for therapeutic purposes, e.g. for a human or animal subject.

"coronavirus" refers to a virus having a corona process on its envelope that is shaped like coronas. Coronavirus particles are irregular in shape and about 60-220nm in diameter. The virion is surrounded by a fat membrane, the surface of which has three glycoproteins: spike Protein (S, Spike Protein, which is the receptor binding site, cytolytic and major antigenic site); envelope proteins (E, Envelope Protein, smaller, Envelope-binding proteins); membrane proteins (M, Membrane Protein, responsible for transmembrane transport of nutrients, budding release of nascent viruses and formation of viral envelope).

The traditional Chinese medicine essential oil can reduce the combination of spike protein and angiotensin converting enzyme 2(ACE2) by reducing the spike protein of coronavirus, thereby playing a role in inhibiting coronavirus. Thus, in one embodiment, the essential oil of Chinese herbs inhibits coronavirus by reducing spike protein of coronavirus coat.

In some embodiments, the coronavirus is severe acute respiratory syndrome coronavirus No. 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV), middle east respiratory syndrome coronavirus (MERS-CoV), or bat SARS-like coronavirus (bat-SL-CoVZC 45).

In some embodiments, the bacteria are one or more of Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa), Klebsiella pneumoniae (Klebsiella pneumoniae), Bacillus Subtilis (BS), Aeromonas hydrophila (Aeromonas hydrophylla), Edwardsiella tarda (edwards tarda), Vibrio alginolyticus (Vibrio alginolyticus), Vibrio harveyi (Vibrio harveyi).

According to a third aspect of the invention, the invention also provides the use of the essential oil of the traditional Chinese medicine of the first aspect of the invention in the preparation of a preparation for preventing coronavirus infection and/or bacterial infection.

In some embodiments, the bacteria is one or more of staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, klebsiella pneumoniae, bacillus subtilis, aeromonas hydrophila, edwardsiella tarda, vibrio alginolyticus, and vibrio harveyi.

In some embodiments, the article of daily use is a mask, a disinfectant, an aromatherapy, a spray, a hand sanitizer, or a skin care product.

In some embodiments, the daily necessities are masks, and the herbal essential oil is present in the masks in the form of medicated thread.

In some embodiments, the concentration of each of the extracts in the herbal essential oils is 5% to 10%, e.g., 5%, 6%, 7%, 8%, 9%, or 10%, or any number in a range of any two of them.

In some embodiments, the mask is a disposable mask comprising a left band, an outer layer as a water barrier layer, a middle layer as a filter layer, an inner layer as a water absorbing layer, a metal strip for clamping the bridge of the nose, and a right band, and the medicated thread is located between the outer layer and the middle layer, e.g., below the metal strip, e.g., 0.5-2 cm below.

In one embodiment, the medicated thread is located between the outer layer and the middle layer and below the metal strip.

In one embodiment, the medicated thread is located 0.5-2 centimeters below the metal strip, such as 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 centimeters below or any distance therebetween.

As is generally understood by those skilled in the art, the left and right straps are primarily for the purpose of wear. The outer layer is mainly used for waterproof, the material is non-woven fabric, and the non-woven fabric has a water-repelling function and can prevent spray and large-particle dust. The middle layer is mainly used for filtering, the material is melt-blown non-woven fabric, and bacteria and particles can be filtered through the melt-blown netting technology. The inner layer is mainly used for absorbing water, is made of non-woven fabric, and is subjected to surface hydrophilic treatment, so that the contact part of the skin is comfortable. The metal strip in the mask is used for clamping the nose bridge, and the material of the metal strip can be aluminum, an environment-friendly plastic strip and the like. The medicated thread is any material with adsorption effect soaked with Chinese medicinal essential oil, such as cotton thread, non-woven thread, cotton sliver or non-woven fabric strip. Further, the length of the medicated thread may be adjusted according to the mask, for example, 17 cm. When the mask is manufactured, the material with adsorption effect is added into the essential oil of traditional Chinese medicine by soaking method to prepare medicated thread, and then the medicated thread is put into a production machine, so that the medicated thread is placed in the mask. Of course, any material known in the art that is commonly used for the above-described corresponding structures of the mask may be used in the present invention, and is not further limited herein.

In the present invention, the mask may be any mask known in the art, and may be a medical mask or a non-medical mask, and may be, for example, an N95 type mask, a medical surgical mask, or the like. Therefore, the medicated thread can be placed at a suitable position according to the present invention according to different mask types.

In the invention, the essential oil is absorbed by soaking the material with adsorption in the traditional Chinese medicine essential oil, and then the medicated thread containing the essential oil is added between the outer waterproof layer and the middle filter layer, preferably at the position below the metal strip, so that on one hand, the medicated thread containing the traditional Chinese medicine essential oil can be prevented from directly contacting the waterproof layer and the filter layer, the effect of the mask on blocking bacteria and viruses can not be influenced, and on the other hand, the medicated thread containing the traditional Chinese medicine essential oil is added in the mask instead of on the surface of the mask, and the medicated thread can be prevented from falling off and losing the effect. In addition, the added medicated thread can be suitable for a full-automatic mask production line.

The invention has the beneficial effects that:

the invention uses the extracts of a plurality of Chinese medicinal materials to prepare the Chinese medicinal essential oil with the effects of resisting bacteria, viruses and the like, thereby being used for inhibiting coronavirus and/or bacteria and preventing and/or treating coronavirus infection and/or bacterial infection. The essential oil of Chinese traditional medicine provided by the invention not only contains the extract of Chinese traditional medicine, for example, by supercritical CO₂The extract of Chinese medicinal materials obtained by extraction or steam distillation, and base oil commonly used in essential oil preparation, can not only give fragrant smell, but also have antibacterial and antiviral effects.

The traditional Chinese medicine essential oil is also applied to daily necessities such as masks, disinfectant, aromatherapy, spray, hand sanitizer or skin care products, has antibacterial and antiviral effects and aromatic taste, can make people happy, and can keep physical and mental health while helping people prevent virus invasion and enhancing upper respiratory tract defense capacity.

The invention applies the traditional Chinese medicine essential oil to the mask in the form of the medicated thread, the medicated thread is prepared by soaking a material with adsorption in the traditional Chinese medicine essential oil, and then the medicated thread is placed between the outer waterproof layer and the middle filtering layer, preferably, the mask is prepared at the position below the metal strip, so that on one hand, the traditional Chinese medicine essential oil can be prevented from directly contacting the waterproof layer and the filtering layer, the effect of blocking bacteria and viruses of the mask can not be influenced, on the other hand, the medicated thread containing the traditional Chinese medicine essential oil is placed in the mask instead of on the surface of the mask, and the effect loss of the medicated thread due to the falling can also be avoided. The mask provided by the invention not only adds various fragrant and fresh smells to the mask, but also has the functions of refreshing, resuscitation, antibiosis, antivirus and the like by adding the medicated thread containing the traditional Chinese medicine essential oil into different layers of the mask on the premise of not damaging the waterproof layer structure of the mask. The mask provided by the invention is simple to manufacture, low in cost, convenient to use and easy to popularize, and the medicated thread added in the mask is also suitable for a full-automatic mask production line.

The present invention will now be described more specifically and in detail with reference to the accompanying drawings and examples, which are only preferred embodiments of the invention and are not intended to limit the invention. All the raw materials and reagents of the invention are conventional market raw materials and reagents unless otherwise specified.

Example 1: preparation method of traditional Chinese medicine essential oil

Preparation of Chinese medicine extract

1. Supercritical CO₂Extraction method

Pulverizing Chinese medicinal materials, adding into an extractor, adding a small amount of polar solvent (carrying agent) into the extractor, and allowing CO to flow at room temperature and low pressure (20-30 deg.C, 100-₂Supercritical fluid reaches subcritical fluid state, fluid is injected into the extractor from bottom to top to enter an extraction stage, after extraction is completed, extract liquor is decompressed and then enters a first separator and a second separator in sequence, and traditional Chinese medicine extract is collected at the bottom of the second separator.

2. Steam distillation process

Placing the pulverized Chinese medicinal materials into a distiller, connecting a cooler at the upper part, and introducing steam at the lower part to maintain the steam pressure of 0.12-0.13 MPa. Vapor permeates the medicinal powder, volatile components in the medicinal powder are gasified due to the high temperature of the vapor, the vapor enters a cooler along with water vapor from a distillation tube, the vapor and the gasified volatile components are cooled in the cooler to form an oil-water mixture, the oil-water mixture is collected at the outlet of the cooler by an oil-water separator, after standing, oil and water are automatically separated, the upper layer is the traditional Chinese medicine extract, and the lower layer is water. Removing water from the lower layer with a separator to obtain the desired Chinese medicinal extract.

Second, preparation of Chinese medicine essential oil

In the following examples supercritical CO will be passed₂Mixing the Chinese medicinal extract and base oil at required ratio to obtain Chinese medicinal essential oil.

Example 2: research on the action against the New coronavirus (SARS-CoV-2) (first experimental principle):

the antiviral efficacy of the herbal extract was demonstrated by examining the inhibition of Spike protein (Spike protein) present in the capsid of the New coronavirus (SARS-CoV-2) by the herbal extract samples. When the spike protein is inhibited by herbal extracts, its binding to the human receptor angiotensin converting enzyme 2(ACE2) is reduced. The less the spike protein binds to ACE2, the stronger the Chinese medicinal extract has the effect of inhibiting the spike protein. The method is an effective in vitro detection means for testing the efficacy of the medicine for resisting the new coronavirus (SARS-CoV-2).

(II) Experimental materials:

the used traditional Chinese medicine essential oil comprises the following components: clove, eupatorium, sandalwood, cablin potchouli herb, zedoary, honeysuckle, folium artemisiae argyi, cinnamon, fructus forsythiae, liquorice, dandelion, perilla leaf, bay leaf, rhizoma atractylodis, radix bupleuri, nutmeg, magnolia flower, citronella, houttuynia cordata, fructus cnidii and the like; SARS-CoV-2Spike ACE2 Inhibitor Screening Assay Kit (from BPS Bioscience) was tested against the New coronaviruses (SARS-CoV-2).

(III) experiment steps:

the spike protein solution at a concentration of 1. mu.g/mL was added to a specially prepared 96-well plate at 50. mu.L per well and incubated overnight at 4 ℃. After incubation was complete, the supernatant was decanted and washed 3 times with 100 μ L of blocking solution a. The solution was patted dry, 100. mu.L of blocking solution B was added to each well, and the mixture was shaken at room temperature for 1 hour. After the solution was patted dry, 20. mu.L of blocking solution A was added to each well, 10. mu.L of 5% DMSO aqueous solution was added to each of the positive control well and the negative control well, 10. mu.L of sample solution (containing 5% DMSO) was added to each of the sample wells, and the mixture was shaken at room temperature for 1 hour. Then, 20 μ L of blocking solution a was added to the positive control wells, the highest inhibition rate of the simulation experiment was performed by adding blocking solution a containing no ACE2-His to the positive control wells, 20 μ L of ACE2-His solution with a concentration of 2.5ng/μ L was added to the negative control wells and the sample wells, and the reaction was performed with shaking at room temperature for 1 hour. After completion of the reaction, the liquid was patted dry and washed 3 times with 100. mu.L of blocking solution A. The solution was patted dry, 100. mu.L of blocking solution B was added to each well, and the mixture was incubated at room temperature for 10 minutes. After the incubation, the liquid was patted dry, 10. mu.L of Anti-His-HRP solution was added to each well, and the mixture was incubated at room temperature with shaking for 1 hour. The cells were washed 3 times with 100. mu.L of blocking solution A, and then 100. mu.L of blocking solution B was added to each well, followed by incubation at room temperature for 10 minutes. Finally, the liquid was patted dry and a mixture of 100. mu. L ELISA ECL substrate A and B (1:1) was added to each well to measure the intensity of chemiluminescence. Data were statistically analyzed with SPSS 22 and significance was expressed as p <0.05 and p < 0.01.

(IV) experimental results:

as shown in fig. 1 and 2, both the low concentration Chinese medicinal essential oil (0.1% by volume) and the high concentration Chinese medicinal essential oil (1% by volume) have an effect of resisting new crown virus (SARS-CoV-2), and the antiviral effects of clove, eupatorium, curcuma zedoary, honeysuckle, cinnamon, forsythia, licorice, dandelion, perilla leaf, bupleurum, magnolia flower, citronella and cnidium fruit are significant. Specifically, the inhibition effect of the low-concentration Chinese medicinal essential oil of eupatorium, honeysuckle, dandelion, houttuynia cordata and fructus cnidii on the combination of the spike protein and the ACE2 is over 90%, wherein the inhibition effect of the eupatorium, the dandelion, the houttuynia cordata and the fructus cnidii on the combination of the spike protein and the ACE2 can almost reach 100% (figure 1), and the inhibition effect of the high-concentration Chinese medicinal essential oil of clove, eupatorium, honeysuckle, liquorice, dandelion, perilla leaf, radix bupleuri and fructus cnidii on the combination of the spike protein and the ACE2 can almost reach 100% (figure 2). In FIGS. 1 and 2, positive controls were 5% DMSO in water and ACE 2-His-free solutions; negative controls were 5% DMSO in water and 2.5 ng/. mu.L ACE2-His in water.

Of these essential oils, we further selected the essential oils of clove, eupatorium, honeysuckle and cinnamon with significant antiviral activity, and measured their anti-new crown virus (SARS-CoV-2) activities at concentrations of 0.003%, 0.01%, 0.03%, 0.1%, 0.3% and 1% (by volume), and the results are shown in fig. 3. As can be seen from FIG. 3, at a concentration of 0.003%, the clove, eupatorium, honeysuckle and cinnamon essential oil also show antiviral ability, and the antiviral activity is in direct proportion to the concentration.

(V) preliminary conclusion:

screening 20 Chinese medicinal extracts for resisting new crown virus (SARS-CoV-2), and finding that the three Chinese medicinal extracts have insignificant antiviral effects, and seventeen extracts have good antiviral effects, wherein the Chinese medicinal essential oils of clove, herba Eupatorii, rhizoma Curcumae, flos Lonicerae, cortex Cinnamomi, fructus forsythiae, radix Glycyrrhizae, herba Taraxaci, folium Perillae, radix bupleuri, flos Magnoliae, herba Cymbopogonis Citrari, fructus Cnidii, etc. all have better effect of resisting new crown virus (SARS-CoV-2). The inhibition effect of patchouli, sandalwood and atractylodes rhizome oil on S protein is not obvious.

Example 3: antibacterial action of Chinese medicinal essential oil (I) experimental strain:

including 5 common airborne pathogens: staphylococcus Aureus (SA); escherichia Coli (EC); pseudomonas Aeruginosa (PA); klebsiella Pneumoniae (KP); bacillus Subtilis (BS), and 4 common aquaculture common pathogens: aeromonas hydrophila (Ah); edwardsiella tarda (edwards siella tarda, Et); vibrio algicidal (Vibrio algicidalicus, Va); vibrio harveyi (Vibrio harveyi, Vh). All bacteria were purchased from the Guangdong province Collection of microorganisms.

(II) culture medium:

5 kinds of aerogenic pathogenic bacteria (SA, EC, PA, KP and BS), Ah and Et were cultured in LB medium at 37 ℃; va and Vh were cultured at 30 ℃ in 2216E medium.

(III) Experimental method:

and (3) determining the antibacterial activity of the extract sample by adopting a filter paper sheet method, and testing the size of the inhibition zone of the sample on bacteria.

(IV) experimental steps:

SA, EC, PA, KP, BS, Ah and Et were prepared in 100. mu.L of 1X 10⁵Respectively coating the CFU/mL suspension on LB agar culture medium; va and Vh were prepared as 1X 10 in 100. mu.L⁵CFU/mL suspension was spread on 2216E agar medium, respectively. Placing filter paper sheets (diameter 6mm) soaked with 20 kinds of Chinese medicinal essential oil on the surface of the culture medium respectively, allowing the filter paper sheets to cling to the surface of the culture medium, standing for 30 minutes, placing in an incubator, culturing under appropriate conditions for 24 hours, and measuring the diameter of the transparent antibacterial ring. Because the traditional Chinese medicine essential oil is in spherical diffusion in the culture medium, the concentration of the traditional Chinese medicine essential oil is decreased from inside to outside, the diameter of the formed transparent ring is in positive correlation with the antibacterial activity.

(V) experimental results:

the 20 kinds of Chinese medicinal essential oil have different growth inhibition effects on nine kinds of bacteria, wherein the cinnamon extract has the most remarkable antibacterial activity which is stronger than that of a positive control kanamycin (5 mg/mL). Specific antimicrobial activities are shown in table 1.

TABLE 1 antibacterial Activity results of twenty Chinese medicinal essential oils

Note: and SA: staphylococcus aureus bacteria; EC: e.coli; KP: klebsiella pneumoniae; PA: pseudomonas aeruginosa; BS: b, bacillus subtilis; ah: aeromonas hydrophila; et: edwardsiella tarda; va: vibrio alginolyticus; and Vh: vibrio harveyi

(VI) preliminary conclusion:

the results of the antibacterial experiments in table 1 show that the extracts of cinnamon, clove and bay leaf have better inhibition effect on 9 bacteria; the citronella extract shows better inhibitory activity to 8 bacteria; the curcuma zedoary extract also shows relatively good bacteriostatic efficacy on 5 bacteria, wherein the cinnamon extract has the most remarkable activity, and the inhibitory activity on seven bacteria is stronger than that of a positive control kanamycin (5 mg/mL).

The inhibitory effects of cinnamon, clove, zedoary, bay leaf and citronella on various bacteria are shown in fig. 4.

Example 4: determination of skin irritation and sensitivity of Chinese medicinal essential oil

Skin sensitivity test: 12 volunteer subjects (5 male and 7 female, 22-45 years old) were treated by diluting the essential oil (grape seed oil as base oil) to 5% (by volume), applying to the inner side of the left arm of the subject, and recording whether there was any sensitive reaction such as redness and itching after 1 hr. The observation results show that: the skin sensitivity phenomena such as red swelling, itching, desquamation, eczema and the like do not appear.

Example 5: manufacture of antiviral essential oil mask

According to the skin test of example 4, no allergic result was found, and we diluted the essential oil of chinese traditional medicine with grape seed oil to 5% (volume ratio), thereby preparing the essential oil of chinese traditional medicine ready for use. Then, the antiviral essential oil mask of the present invention is further manufactured on the basis of a general mask.

The first method is as follows: adding 10 mu L of instant Chinese medicinal essential oil into the non-woven cotton stuck with double-sided adhesive tape until the essential oil is completely adsorbed, and sticking the essential oil on the bottommost part of the back surface of the disposable mask, thereby preparing the antiviral essential oil mask.

The second method comprises the following steps: the medicated thread 2 is obtained by impregnating cotton thread, cotton sliver or non-woven fabric strip having adsorption function with the essential oil of the Chinese medicinal materials of the present invention, and then fixed at an upper position between the outer layer 3 (i.e., waterproof layer) and the middle layer 4 (i.e., filter layer) of the disposable mask, for example, at about 1 cm below the metal thread 1 sandwiching the bridge of the nose, to facilitate nasal mucosa absorption (fig. 5). Another antiviral essential oil mask of the present invention was obtained in the above manner.

It should be noted that it is not desirable to add essential oil directly to the waterproof layer of the mask because the essential oil molecules can damage the water layer and the electrostatic filter layer, rendering the mask ineffective. In addition, the structural configuration can avoid direct contact with the waterproof layer and the filter layer, and does not influence the effect of the mask on blocking viruses and bacteria; in addition, the medicated thread is placed in the mouth cover instead of on the surface, so that the medicated thread can be prevented from falling off and losing efficacy.

Example 6: application of antiviral essential oil mask

14-20 volunteer subjects were assigned to prevent inhalation of new coronaviruses. Mask and questionnaire survey with different essential oils of Chinese medicinal materials are distributed every day. The statistical results are shown in table 2 below, and table 3 shows the contents of the questionnaire: gender, age, wearing time, smell, taste duration, satisfaction, discomfort to the eyes, nose and skin of a human, etc.

TABLE 2 questionnaire statistics after wearing disposable mask with essential oils of traditional Chinese medicine

TABLE 3 questionnaire details of eight Chinese medicinal essential oils

And (4) conclusion: as shown by the statistical results in tables 2 and 3, the most satisfactory essential oil of Chinese traditional medicine is Eupatorium. Additionally, cinnamon oil and bupleurum oil are combined in a 1:1 ratio; and clove oil and licorice oil and eupatorium oil and bupleurum oil in a 1:1 combination, all gave better satisfaction in questionnaires.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims

1. A traditional Chinese medicine essential oil, wherein the traditional Chinese medicine essential oil comprises one or more of the following traditional Chinese medicine extracts: clove, cinnamon, curcuma zedoary, perilla leaf, magnolia flower, citronella, fructus cnidii, eupatorium, liquorice, honeysuckle, fructus forsythiae, dandelion and radix bupleuri.

2. The essential oil of Chinese herbs of claim 1, wherein the extract of Chinese herbs is extracted by supercritical CO₂Extracting with extraction method or steam distillation method.

3. The herbal essential oil of claim 1 or 2, wherein the herbal essential oil comprises a combination of the following herbal extracts: zedoary extract + cinnamon extract, bupleurum extract + cinnamon extract, licorice extract + clove extract, or eupatorium extract + bupleurum extract.

4. The essential oil of any one of claims 1 to 3, further comprising one or more of sweet almond oil, rose seed oil or grape seed oil as a base oil.

5. The herbal essential oil according to any one of claims 1 to 4, wherein the concentration of each extract in the herbal essential oil is between 0.003% and 10% by volume.

6. The traditional Chinese medicine essential oil according to claim 5, wherein the concentration of each extract in the traditional Chinese medicine essential oil is 0.1-10% by volume percentage.

7. A method for inhibiting coronavirus and/or bacteria, said method comprising contacting an essential oil of a Chinese medicinal material according to any one of claims 1 to 6 with coronavirus and/or bacteria.

8. The method of claim 7, wherein the coronavirus is severe acute respiratory syndrome coronavirus No. 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV), middle east respiratory syndrome coronavirus (MERS-CoV), or bats SARS-like coronavirus (bat-SL-CoVZC 45).

9. The method of claim 7, wherein the bacteria is one or more of Staphylococcus aureus (Staphyloccus aureus), Escherichia coli (Escherichia coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa), Klebsiella pneumoniae (Klebsiella pneumoniae), Bacillus Subtilis (BS), Aeromonas hydrophila (Aeromonas hydrophylla), Edwardsiella tarda (Edwardsiella tarda), Vibrio alginolyticus (Vibrio algirylicus), and Vibrio harveyi (Vibrio harveyi).

10. The method of any one of claims 7-9, wherein the chinese medicinal essential oil inhibits coronavirus by reducing spike protein of coronavirus coat.

11. Use of the essential oil of a Chinese medicinal material according to any one of claims 1 to 6 in the manufacture of a formulation for the prevention of coronavirus infection and/or bacterial infection.

12. The use of claim 11, wherein the coronavirus is severe acute respiratory syndrome coronavirus No. 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV), middle east respiratory syndrome coronavirus (MERS-CoV), or bats SARS-like coronavirus (bat-SL-CoVZC 45).

13. The use according to claim 11, wherein the bacteria is one or more of staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, klebsiella pneumoniae, bacillus subtilis, aeromonas hydrophila, edwardsiella tarda, vibrio alginolyticus, and vibrio harveyi.

14. A daily necessity comprising the essential oil of the Chinese medicine according to any one of claims 1 to 6.

15. The daily necessity of claim 14, wherein the daily necessity is a mask, a disinfectant, an aromatherapy, a spray, a hand sanitizer, or a skin care product.

16. The daily necessity according to claim 15, wherein the daily necessity is a mask, and the essential oil of the chinese medicine is present in the mask in the form of medicated thread.

17. The daily item of claim 16, wherein the concentration of each of the extracts in the herbal essential oils is 5% -10%, such as 5%, 6%, 7%, 8%, 9% or 10%.

18. A commodity according to claim 16 or 17, wherein the mask is a disposable mask comprising a left band, an outer layer as a waterproof layer, a middle layer as a filter layer, an inner layer as a water absorbing layer, a metal strip for clamping the bridge of the nose and a right band, and the medicated thread is located between the outer layer and the middle layer, such as below the metal strip, such as 0.5-2 cm below.