Traditional Chinese medicine material extract, preparation method and use thereof

Abstract

A traditional Chinese medicine extract for treating coronavirus-related symptoms or diseases, and a preparation method and use thereof. The traditional Chinese medicine extract is made of eaglewood, soapberry, horn thorn, cinnamon leaf and camphor leaf.

Description

Technical Field

[0001] This invention relates to the technical field of extracts from traditional Chinese medicinal materials, and more particularly to an extract from traditional Chinese medicinal materials, its preparation method, and its uses. Background Technology

[0002] Coronaviruses are zoonotic RNA viruses that can spread between animals and humans. Under an electron microscope, they appear spherical or oval in shape. They are called coronaviruses because of their crown-like appearance with protruding cystic collagen fibers. The genome size of coronaviruses ranges from 26,000 to 32,000 base pairs, making them the largest class of RNA viruses in terms of genome size.

[0003] The novel coronavirus SARS-CoV-2 (initially reported as COVID-19, and officially named SARS-CoV-2 by the Coronavirus Research Group based on phylogenetic relationships, taxonomy, and convention) is a new viral strain. SARS-CoV-2 binds to human cell surface receptors using its viral spike protein and produces various important proteins within the virus through the 3CLpro protease, which is cleaved from the protein chain transcribed from the viral gene, allowing the virus to continue infecting and replicating.

[0004] As the novel coronavirus epidemic rages, common signs of novel coronavirus infection include respiratory symptoms, fever, cough, shortness of breath or difficulty breathing, and in more severe cases, even death. Therefore, there is an urgent need for drugs that can be used to treat coronavirus-related symptoms or diseases. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides a Chinese herbal extract, its preparation method, and its uses. By taking a medicine containing the Chinese herbal extract, the efficacy of treating coronavirus-related symptoms or diseases can be achieved.

[0006] This invention provides a Chinese herbal extract comprising an active ingredient, which is composed of any one or a combination of agarwood, soapberry, horn thorn, cinnamon leaf, and camphor leaf.

[0007] In another embodiment of the present invention, when the effective ingredient is composed of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf, the weight ratio of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf is 1-5:1-5:1-5:1-5:1-5.

[0008] In another embodiment of the present invention, the weight ratio of the agarwood, the soapberry, the thorn, the cinnamon leaf, and the camphor leaf is 1:1:1:1:1.

[0009] Another embodiment of the present invention provides a method for preparing a traditional Chinese medicine extract, which includes the following steps: an extraction step: mixing a raw material containing any one or a combination of agarwood, soapberry, thorn, cinnamon leaf, and camphor leaf with ultrapure water at a weight ratio of 1:5 to 1:10, and heating to extract an extract, wherein the extraction temperature is between 90°C and 110°C, and the extraction time is between 1.5 hours and 2.5 hours; a filtration step: filtering the extract; and a drying step: performing vacuum freeze-drying on the extract to obtain the traditional Chinese medicine extract.

[0010] In another embodiment of the present invention, when the raw material is composed of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf, the weight ratio of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf is 1-5:1-5:1-5:1-5:1-5.

[0011] In another embodiment of the present invention, the weight ratio of the agarwood, the soapberry, the thorn, the cinnamon leaf, and the camphor leaf is 1:1:1:1:1.

[0012] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extract in the preparation of a medicament for treating coronavirus-related symptoms or diseases, wherein the active ingredient of the Chinese herbal extract is cinnamon leaf.

[0013] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extract in the preparation of a drug that intervenes in the spike protein of SARS-CoV-2, wherein the active ingredient of the Chinese herbal extract is cinnamon leaf.

[0014] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extract in the preparation of a drug that inhibits the activity of SARS-CoV-23CLpro protease, wherein the active ingredient of the Chinese herbal extract is cinnamon leaf.

[0015] In another embodiment of the present invention, the half-inhibitory concentration (IC50) of the agent is... 50 The value ranges from 10 to 15 μg / mL.

[0016] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extracts in the preparation of a medicament for treating coronavirus-related symptoms or diseases.

[0017] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extracts in the preparation of a drug that intervenes in the spike protein of SARS-CoV-2.

[0018] In another embodiment of the present invention, the half-effective concentration EC of the agent is... 50 The value ranges from 20 to 30 μg / mL.

[0019] Another embodiment of the present invention provides the use of the above-mentioned Chinese herbal extracts in the preparation of a drug that inhibits the activity of SARS-CoV-23CLpro protease.

[0020] In another embodiment of the present invention, the half-inhibitory concentration (IC50) of the agent is... 50 The value ranges from 26 to 40 μg / mL.

[0021] In another embodiment of the present invention, the use of the Chinese herbal extract includes the dosage form of the medicine, which includes mouthwash, nasal inhaler, or oral capsule.

[0022] Therefore, by taking medicines containing extracts of traditional Chinese medicine, it is possible to effectively intervene in the binding of the spike protein of the coronavirus to human angiotensin-converting enzyme 2 (hACE2) on the surface of human cells, and inhibit the activity of the coronavirus 3CLpro protease, thereby achieving the effect of treating coronavirus-related symptoms or diseases. Attached Figure Description

[0023] Figure 1 This is a flowchart illustrating the preparation process of the extract from the Chinese medicinal materials of this invention.

[0024] Figure 2 This is the result of the intervention analysis of the Chinese herbal extracts in the first embodiment of the present invention on the binding of SARS-CoV-2 spike protein to in vitro synthesized hACE2.

[0025] Figure 3 The IC50 of the herbal extract of the first embodiment of the present invention is the half-maximal inhibitory concentration (IC50) of the inhibitory effect on the binding of SARS-CoV-2 spike protein to in vitro synthesized hACE2. 50 Analysis results.

[0026] Figure 4 The half-effective concentration (EC50) of the herbal extract in the first embodiment of this invention against the binding of SARS-CoV-2 pseudovirus to SmBiT-ACE2-expressing cells is... 50 Analysis results.

[0027] Figure 5 This is the analysis result of the Chinese herbal extracts from the first embodiment of the present invention inhibiting the activity of SARS-CoV-2 3CLpro protease.

[0028] Figure 6 The half-maximal inhibitory concentration (IC50) of the herbal extract in the first embodiment of this invention against the SARS-CoV-2 3CLpro protease activity is... 50 Analysis results.

[0029] Figure 7 The half-maximal inhibitory concentration (IC50) of the herbal extracts in the second embodiment of this invention against the SARS-CoV-2 3CLpro protease activity is... 50 Analysis results.

[0030] Wherein, S1: extraction step; S2: filtration step; S3: drying step. Detailed Implementation

[0031] To facilitate the explanation of the central idea expressed in the above-mentioned description of the invention, specific embodiments are used.

[0032] This invention provides a Chinese herbal extract and its preparation method. The Chinese herbal extract contains an active ingredient. The active ingredient is composed of any one of the following materials, or a combination thereof: agarwood, soapberry, horn thorn, cinnamon leaf, and camphor leaf.

[0033] In one embodiment of the present invention, the active ingredient is any one of the following single-herb Chinese medicines: agarwood, soapberry, thorn, cinnamon leaf, and camphor leaf.

[0034] In one embodiment of the present invention, the effective ingredient is any two of the following compound Chinese herbal medicines: agarwood, soapberry, horn thorn, cinnamon leaf, or camphor leaf, selected from any one combination of agarwood and soapberry, agarwood and horn thorn, agarwood and cinnamon leaf, agarwood and camphor leaf, soapberry and horn thorn, soapberry and cinnamon leaf, soapberry and camphor leaf, horn thorn and cinnamon leaf, horn thorn and camphor leaf, and cinnamon leaf and camphor leaf.

[0035] In one embodiment of the present invention, the effective ingredient is any three of the following compound Chinese herbal medicines: agarwood, soapberry, thorn, cinnamon leaf, or camphor leaf. For example, a combination of agarwood, soapberry, and thorn; a combination of agarwood, soapberry, and cinnamon leaf; or a combination of agarwood, soapberry, and camphor leaf.

[0036] In one embodiment of the present invention, the active ingredient is any four of the following compound Chinese herbal medicines: agarwood, soapberry, thorn, cinnamon leaf, or camphor leaf. For example, a combination of agarwood, soapberry, thorn, and cinnamon leaf, or any combination of soapberry, thorn, cinnamon leaf, and camphor leaf.

[0037] In one embodiment of the present invention, the effective ingredients are composed of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf. The weight ratio of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf is preferably 1-5:1-5:1-5:1-5:1-5:1-5. In this embodiment, the weight ratio of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf is 1:1:1:1:1.

[0038] Materials and material handling methods:

[0039] 1. Materials

[0040] (a) Agarwood

[0041] Agarwood is derived from the resinous wood of *Aquilaria sinensis* or *Aquilaria malaccensis*, both dicotyledonous plants belonging to the Thymelaeaceae family. Agarwood possesses properties such as promoting qi circulation and relieving pain, regulating the middle jiao (digestive system), warming the kidneys and promoting qi absorption, clearing the meridians and calming the mind, and sedation and analgesia. Furthermore, pharmacological studies have confirmed that agarwood exhibits significant antibacterial activity. In this embodiment, the agarwood wood is selected from *Aquilaria sinensis* from Vietnam or Taiwan. In other possible embodiments, *Aquilaria crassna*, *Aquilaria malaccensis*, or *Aquilaria yunnanensis* can also be used.

[0042] (b) Soapberry

[0043] Soapberry is derived from the mature pods of the legume plant Gleditsia sinensis. Soapberry has the effects of relieving stubborn phlegm, clearing the orifices, dispelling wind and killing insects. Soapberry contains triacanthin, which has an inhibitory effect on pathogens pharmacologically.

[0044] (c) horns

[0045] Horn thorns are taken from the thorns of the legume plant Gleditsia sinensis. Horn thorns have the effects of reducing swelling, drawing out toxins, draining pus, and killing insects. Flavonoids, phenols, and amino acids can be extracted from horn thorns. Horn thorn extract has antibacterial and disinfectant effects in pharmacology.

[0046] (d) Cinnamon leaves

[0047] Cinnamon leaves are derived from the plant Cinnamomum osmophloeum, belonging to the Lauraceae family. They have the effects of dispelling cold and relieving pain, promoting blood circulation, and strengthening the stomach. Pharmacologically, cinnamon leaf extract can inhibit bacterial growth, thereby achieving the effect of antibacterial disinfection.

[0048] (e) Camphor leaves

[0049] Camphor leaves are taken from the camphor tree (Cinnamomum camphora), a plant of the Lauraceae family. They have the effects of dispelling wind, removing dampness, detoxifying, and killing insects. Camphor leaf extract has antibacterial and disinfecting effects in pharmacology.

[0050] 2. Material handling methods:

[0051] (a) Wash the agarwood, soapberry, thorn, cinnamon leaves and camphor leaves separately and let them air dry overnight.

[0052] (b) Next, the agarwood, soapberry, thorn, cinnamon leaf and camphor leaf are dried in an oven. The moisture content of the agarwood, soapberry, thorn, cinnamon leaf and camphor leaf after drying is preferably between 5% and 20%, and the optimal moisture content is 10%.

[0053] (c) Grind agarwood, soapberry, thorn, cinnamon leaf and camphor leaf separately using a grinder. The fineness of the ground agarwood, soapberry, thorn, cinnamon leaf and camphor leaf should be between 10 mesh and 30 mesh, with 20 mesh being the best.

[0054] Please refer to the following: Figure 1 The figure shows a method for preparing the herbal extract of the present invention, which includes the following steps:

[0055] Extraction step S1: First, a raw material containing any one or a combination of agarwood, soapberry, thorn, cinnamon leaf, and camphor leaf is mixed with ultrapure water at a weight ratio of 1:5 to 1:10, wherein a weight ratio of 1:10 is preferred. After mixing the raw material and ultrapure water, the mixture is placed in a liquid nitrogen tank and frozen, then thawed at room temperature to perform a freeze-thaw cycle to break the cell walls of the raw material. Next, the raw material and ultrapure water are heated for extraction to obtain an extract, wherein the extraction temperature is between 90℃ and 110℃, preferably 100℃, and the extraction time is between 1.5 hours and 2.5 hours, preferably 2 hours.

[0056] Filtration step S2: After the extract has cooled, it is filtered. In this embodiment, the filtration is performed in four steps to obtain the filtered extract. First, the extract is coarsely filtered. Then, the coarsely filtered extract is centrifuged. After centrifugation, the supernatant is collected. The centrifugation speed is 7,000-15,000 rpm and the centrifugation time is 3-10 minutes. The optimal centrifugation speed is 10,000 rpm and the centrifugation time is 5 minutes. Next, the supernatant is filtered with qualitative filter paper (Whatman NO.4 qualitative filter paper) with a pore size of 15-30 μm. Finally, it is filtered with a glass fiber membrane (MGC glass fiber membrane) with a finer pore size of 1.2 μm to obtain the filtered extract.

[0057] Drying step S3: After the filtration step, the filtered extract is placed in an environment of -35 to -10°C to form a frozen state, and the frozen extract is subjected to vacuum freeze drying to obtain the Chinese herbal medicine extract. The Chinese herbal medicine extract is freeze-dried powder to facilitate preservation and preparation for application.

[0058] In this embodiment, the herbal extract can be prepared into aqueous solutions of different concentrations relative to the solvent. The solvent used in the preparation can be water, reverse osmosis water, deionized water, ethanol, methanol, isopropanol or other suitable solvents with low irritation.

[0059] The following discloses the analytical results of the Chinese herbal extracts according to the first embodiment of the present invention. In this embodiment, the effective components of the Chinese herbal extracts are composed of agarwood, soapberry, thorn, cinnamon leaf, and camphor leaf, with a weight ratio of 1:1:1:1:1. The analytical results include detailed steps and results of the ability of SARS-CoV-2 spike protein to bind and intervene with human angiotensin-converting enzyme 2 (hACE2), as well as the ability to inhibit the activity of SARS-CoV-2 3CLpro protease. All numerical bars presented in the analytical results are the mean ± standard deviation of 50 repeated experiments.

[0060] 1. Analysis of the binding intervention of the full-sequence spike protein of SARS-CoV-2

[0061] The spike protein of SARS-CoV-2 plays an important role in the mechanism of viral entry into human cells. In this embodiment, the potential inhibitory effect and intervention ability of the extract of traditional Chinese medicine of the present invention on the binding of the spike protein of SARS-CoV-2 (Wuhan strain) full sequence to hACE2 were evaluated by enzyme-linked immunosorbent assay (ELISA). The hACE2 used in the experiment was a synthetic protein synthesized in vitro.

[0062] The experimental steps of enzyme-conjugated immunosorbent assay are as follows:

[0063] First, 100 μL of a pricklein solution (500 ng / mL; CAT.GTX135972-pro, GeneTex, Taipei, Taiwan, China) diluted in 15 mM sodium carbonate and 35 mM sodium bicarbonate (pH 9.6) was injected into a 96-well microanalytical disc. The disc was then sealed and incubated overnight at 4°C. Next, the 96-well microanalytical disc containing the pricklein solution was rinsed three times with a rinse buffer consisting of 0.05% (v / v) phosphate buffer and Tween-20 to remove any pricklein solution that could not adhere to the disc.

[0064] Next, 300 μL of the ELISA blocking solution, prepared with a wash buffer containing 0.5% (w / v) fetal bovine serum, was added to a 96-well microanalytical dish and incubated at 37°C for 1.5 hours. After washing three times, 100 μL of different concentrations (0 to 100,000 μg / mL) of the herbal extracts of the present invention or 100 μL of the inhibitor (10 μg / mL; cat. GTX635791, GeneTex, Taipei, Taiwan, China) were added to the 96-well microanalytical dish and incubated at 37°C for 1 hour. The 96-well microanalytical dish was then washed three times with the wash buffer.

[0065] Next, 100 μL of the ELISA signal amplification agent Streptavidin-HRP (100 ng / mL; cat. GTX30949, GeneTex, Taipei, Taiwan, China) was added and the mixture was incubated at 37°C for 1 hour.

[0066] Finally, 200 μL of TMB chromogenic substrate was added to each well of the 96-well microplate and incubated at 37°C for 1 hour in the dark. After the reaction was complete, 50 μL of stop solution was used to terminate the reaction. The absorbance was measured at a wavelength of 450 nm using a microplate reader.

[0067] like Figure 2 The figures show the intervention results of the herbal extracts and inhibitors (RBD antibodies) of this invention on the binding of SARS-CoV-2 spike protein to hACE2. In the absence of hACE2, only the background signal of the SARS-CoV-2 spike protein coating could be measured. The control group had hACE2 added to the SARS-CoV-2 spike protein coating, which produced a strong colorimetric signal with a binding efficiency of 100%. The experimental group showed the intervention effects of adding different concentrations of herbal extracts on the binding of SARS-CoV-2 spike protein to hACE2.

[0068] It can be observed that when the concentration of the added Chinese herbal extract exceeds 10,000 μg / mL, the binding efficiency of SARS-COV-2 spike protein to hACE2 drops to below 50%. Even when the concentration of the Chinese herbal extract is 100,000 μg / mL, the binding efficiency of SARS-COV-2 spike protein to hACE2 drops to 4%, which is comparable to the effect of inhibitors (RBD antibodies) in inhibiting SARS-COV-2 spike protein to hACE2.

[0069] Further nonlinear regression analysis was performed on the binding efficiency of the herbal extract of the present invention against SARS-CoV-2 spike protein and hACE2. After 50 experiments, the half-maximal inhibitory concentration (IC50) of the herbal extract of the present invention was determined. 50 The half-maximal inhibitory concentration (IC50) ranges from 7210 to 10850 μg / mL. 50 The optimal average value is 9024 μg / mL, such as Figure 3 As shown in the figure. The results show that the herbal extract of the present invention effectively inhibits the binding efficiency of spinin and hACE2 in a dose-dependent manner.

[0070] This invention further includes a neutralization test between a Chinese herbal medicine extract and the SARS-CoV-2 pseudovirus. The solvent used was deionized water, and the concentration of the Chinese herbal medicine extract was 250 mg / mL. The test was conducted under the conditions shown in the table below.

[0071]

[0072]

[0073] HeLa expressing SmBiT-ACE2 cells, SARS-COV-2 pseudovirus, and different doses of Chinese herbal extracts were co-cultured for 6 hours, with an equal volume of PBS solution as a control. After 24 hours of culture, the cells were washed twice with PBS solution, and finally, biofluorescence was detected by NanoLuc.

[0074] The test results are shown in the table below. The table shows the neutralization data of Chinese herbal extracts and SARS-CoV-2 pseudovirus. The results show that the minimum dose of Chinese herbal extracts that can block SARS-CoV-2 infection of cells is 50 μg / mL, and the blocking rate is as high as 95%.

[0075]

[0076] In addition, such as Figure 4 As shown, Figure 4 The half-effective concentration (EC50) of the extract of traditional Chinese medicine against the binding of SARS-CoV-2 pseudovirus to HeLa SmBiT-ACE2-expressing cells is given. 50 The analysis results showed that the extracts of traditional Chinese medicine exhibited dose-dependent neutralizing activity against SARS-CoV-2 pseudovirus, with a half-effective concentration (EC50) of 1,000%. 50 The concentration (for 50% of the maximum effect) ranges from 20 to 30 μg / mL, and its half-effective concentration (EC50) is... 50The optimal average value is 24.5 μg / mL.

[0077] 2. Analysis of SARS-CoV-2 3CLpro protease activity inhibition

[0078] SARS-CoV-2 3CLpro protease is an enzyme involved in viral RNA synthesis and replication. In this embodiment, the inhibitory effect of the herbal extract of the present invention on the activity of SARS-CoV-2 3CLpro protease was evaluated by fluorescence resonance energy transfer (FRET) technology.

[0079] The experimental steps for fluorescence resonance energy transfer (FRET) analysis are as follows:

[0080] First, the DABCYL-TSAVLQSGFRKMG-EDANS (Genomics, Taiwan, China) modified peptide was dissolved in water and co-cultured with SARS CoV-2 3CLpro.

[0081] Next, a multi-functional micro-disc analyzer was used. The multimode microplate reader (TECAN) was used as the experimental instrument. The experiment was conducted at 37°C to ensure protein activity. The EDANS fluorescence intensity during the hydrolysis of the modified peptide was used as a function of time. The emission wavelengths were 340 nm and 490 nm, with bandwidths of 9 nm and 15 nm, respectively.

[0082] Next, before testing, it was assessed whether the emission wavelength of the test sample at 340 nm would overlap with the emission spectrum of EDANS. Finally, the effects of different concentrations (0 to 31.25 μg / mL) of the herbal extracts of this invention on the activity of SARS-CoV-2 3CLpro protease were tested, and statistical analysis was performed using GraphPad Prism 7.03 (GraphPad Software, San Diego, CA, USA).

[0083] like Figure 5The figure shows the analytical results of the inhibition of SARS-CoV-2 3CLpro protease activity by the herbal extracts of this invention. It can be observed that after using 0.98 μg / mL of the herbal extract, the relative activity of SARS-CoV-2 3CLpro decreased to approximately 80%; after using 7.81 μg / mL of the herbal extract, the relative activity of SARS-CoV-2 3CLpro decreased to approximately 70%. With the increase of the concentration of the herbal extract to 15.63–31.25 μg / mL, the relative activity of SARS-CoV-2 3CLpro was further inhibited, decreasing to approximately 60% and 50%, respectively.

[0084] After 50 experiments, the half-maximal inhibitory concentration (IC50) of the herbal extract of this invention against the SARS-CoV-2 3CLpro protease activity was determined. 50 The value ranges from 26 to 40 μg / mL, and its half-inhibitory concentration (IC50) is... 50 The optimal average value is 32.61 ± 4.226 μg / mL. Figure 6 As shown, the results indicate that the herbal extract of the present invention can reduce the relative activity of SARS-CoV-2 3CLpro in a dose-dependent manner, demonstrating that the herbal extract of the present invention has therapeutic potential against SARS-CoV-2.

[0085] The following discloses the analytical results of the Chinese herbal extract of the second embodiment of the present invention; in this embodiment, the effective component of the Chinese herbal extract is cinnamon leaf; the analytical results are the results of the inhibitory ability of SARS-CoV-2 3CLpro protease activity, and the detailed steps are the same as those described in the first embodiment above, and will not be repeated here. The numerical bars presented in all analytical results are the average value ± standard deviation of 50 repeated experiments.

[0086] like Figure 7 As shown, after 50 experiments, the half-maximal inhibitory concentration (IC50) of the herbal extract of this invention against the SARS-CoV-2 3CLpro protease activity was [value missing]. 50 The value ranges from 10 to 15 μg / mL, and its half-inhibitory concentration (IC50) is... 50 The optimal average value was 13.08 ± 0.73 μg / mL.

[0087] It should be noted that, in this embodiment, when the concentration of the herbal extract is 25 μg / mL and 50 μg / mL, it can inhibit 69% and 83% of the SARS-CoV-2 3CLpro protease activity, respectively. When the concentration of the herbal extract is 80 μg / mL, it can inhibit 100% of the SARS-CoV-2 3CLpro protease activity.

[0088] As can be seen from the above examples, cinnamon leaf extract has excellent performance in inhibiting the activity of SARS-CoV-2 3CLpro protease. Therefore, the present invention can enhance and expand the therapeutic effect on coronavirus-related symptoms or diseases by adjusting the formulation of single and compound prescriptions through the pharmacological effects produced by the effective components of different compositions and proportions, thereby achieving the best therapeutic effect.

[0089] For example, if a patient is in the early stages of contracting coronavirus, since the viral load in the patient's body is still high at this time, a formula with a higher weight ratio of cinnamon leaves can be used to effectively inhibit the increase of the virus and reduce the overall efficiency of the virus. The weight ratio of cinnamon leaves to agarwood, soapberry, thorn, and camphor leaves can be 1.5:1:1:1:1, or 2:1:1:1:1, or 2.5:1:1:1:1, or 3:1:1:1:1, or 3.5:1:1:1:1, or 4:1:1:1:1, or 4.5:1:1:1:1, or 5:1:1:1:1.

[0090] As mentioned earlier, agarwood has the effects of promoting qi circulation and relieving pain, regulating the middle jiao, warming the kidneys and promoting qi absorption, clearing the meridians and calming the mind, and sedation and analgesia. Gleditsia sinensis has the effects of removing stubborn phlegm, clearing the orifices and opening the pores, and dispelling wind and killing insects. Gleditsia sinensis has the effects of reducing swelling, drawing out toxins, draining pus, and killing insects. Camphor leaves have the effects of dispelling wind, removing dampness, detoxifying, and killing insects. Therefore, when patients are in the middle and late stages of contracting coronavirus, since the viral load in the patient's body has decreased at this time, a formula with a lower weight ratio of cinnamon leaves and a higher weight ratio of agarwood, soapberry, horn, and camphor leaves can be used to improve effects such as analgesia, expectoration, swelling reduction, and dampness removal.

[0091] In addition, the herbal extract of the present invention can be used to prepare a medicine for treating coronavirus-related symptoms or diseases, wherein the medicine comprises the herbal extract of the present invention and a medically acceptable carrier.

[0092] According to the present invention, the aforementioned "carrier" refers to a drug carrier commonly used in the pharmaceutical field, such as fillers, binders, diluents, excipients, acceptors or the like, which are well known to those skilled in the art of manufacturing pharmaceutical products.

[0093] In this embodiment, the dosage form of the medicine includes mouthwash, nasal / oral inhaler, or oral capsule, but is not limited to these. Specifically, by using mouthwash containing Chinese herbal extracts, residual viruses in the mouth and throat can be bound and removed by spitting; while nasal / oral inhalers containing Chinese herbal extracts can utilize the microvessels in the nasal cavity or lungs to absorb the extracts and eliminate the coronavirus; and by taking oral capsules containing Chinese herbal extracts, the extracts can be directly absorbed and utilized by the body through the stomach to eliminate the coronavirus.

[0094] According to this invention, the dosage of the medicine prepared from the extract of traditional Chinese medicinal materials can be determined by relevant medical personnel based on factors such as the patient's age, weight, health status, and disease progression, in accordance with common knowledge in the technical field. The aforementioned medicine prepared from the extract of traditional Chinese medicinal materials can also be administered alone or in combination with other medicines, and the course of treatment should be carried out according to pharmaceutically routine methods by a physician or relevant personnel.

[0095] Therefore, by taking medicines containing extracts of traditional Chinese medicine, it is possible to effectively intervene in the binding of the spike protein of SARS-CoV-2 coronavirus to hACE2 and inhibit the activity of the 3CLpro protease of SARS-CoV-2 coronavirus, thereby achieving the effect of treating coronavirus-related symptoms or diseases.

[0096] The embodiments described above are merely illustrative of the present invention and are not intended to limit the scope of the invention. All modifications or variations made without departing from the spirit of the present invention are within the scope of protection intended by the present invention.

Claims

1. A traditional Chinese medicine extract for treating coronavirus-related symptoms or diseases, characterized in that, The active ingredients are made from agarwood, soapberry, thorn, cinnamon leaf and camphor leaf in a weight ratio of 1:1:1:1:

1.

2. A method for preparing the extract of traditional Chinese medicinal materials as described in claim 1, characterized in that, It is made by the following steps: Extraction steps: The raw materials of agarwood, soapberry, thorn, cinnamon leaf and camphor leaf and ultrapure water are mixed in a weight ratio of 1:5 to 1:10 and subjected to a freeze-thaw cycle. The mixture is then heated to extract the extract. The extraction temperature is between 90℃ and 110℃ and the extraction time is between 1.5 hours and 2.5 hours. Filtration steps: Filter the extract; and Drying step: The filtered extract is freeze-dried under vacuum to obtain the extract of the Chinese medicinal material.

3. The use of the Chinese herbal extract as described in claim 1 in the preparation of pharmaceutical formulations for treating coronavirus-related symptoms or diseases.

4. The use of the Chinese medicinal herb extract as described in claim 3, characterized in that, IC50 of a drug formulation 50 The value ranges from 10 to 15 μg / mL.

5. The use of the Chinese herbal extract as described in claim 1 in the preparation of pharmaceutical formulations that intervene in the spike protein of SARS-CoV-2.

6. The use of the Chinese medicinal herb extract as described in claim 5, characterized in that, Half-effective concentration (EC) of a drug formulation 50 The value ranges from 20 to 30 μg / mL.

7. The use of the herbal extract as described in claim 1 in the preparation of a pharmaceutical formulation that inhibits the activity of SARS-CoV-2 3CLpro protease.

8. The use of the Chinese medicinal herb extract as described in claim 7, characterized in that, The half-inhibitory concentration (IC50) of this drug formulation 50 The value ranges from 26 to 40 μg / mL.

9. The use of the Chinese medicinal herb extract as described in any one of claims 3, 5, and 7, characterized in that, The dosage forms of this drug include mouthwash, nasal / oral inhaler, or oral capsules.

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