Use of musk in the preparation of drugs against respiratory viruses
By using muscone as the active ingredient to prepare an antiviral drug for respiratory viruses, the problem of the lack of reported effects of musk on respiratory viruses has been solved. This has achieved effective inhibition and prevention of various respiratory viruses, especially significantly reducing viral titers of coronaviruses, and has broad clinical application potential.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SICHUAN CENT FOR TRANSLATIONAL MEDICINE OF TRADITIONAL CHINESE MEDICINE
- Filing Date
- 2022-09-08
- Publication Date
- 2026-07-31
AI Technical Summary
The effects and mechanisms of musk or its active ingredients on respiratory viruses have not been reported in the existing technology, and there is a lack of effective antiviral drugs, especially for coronaviruses, human metapneumoviruses, etc., and there are insufficient treatment and prevention methods.
Using muscone as the active ingredient, a drug for treating and preventing respiratory tract infections is prepared. The formulation is made with pharmaceutically acceptable excipients and is used to treat and prevent respiratory tract infections, including upper and lower respiratory tract infections, against a variety of respiratory viruses such as coronaviruses and human metapneumoviruses. The concentration range is 1–1000 μM, preferably 6.25–880 μM.
Muskone exhibits significant antiviral activity at different concentrations, effectively inhibiting and preventing various respiratory viral infections. In particular, it significantly reduces viral titers at concentrations above 110 μM, showing broad clinical application prospects.
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Figure CN116139113B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedicine, specifically relating to the use of musk in the preparation of drugs against respiratory viruses. Background Technology
[0002] Respiratory viruses are viruses that invade the respiratory tract, proliferate in the respiratory mucosal epithelial cells, and cause local respiratory infections or lesions in tissues and organs outside the respiratory tract. Invasion of the upper respiratory tract can lead to diseases such as pharyngitis and sinusitis, while invasion of the lower respiratory tract can cause lung infections, such as respiratory viral pneumonia. Common respiratory viruses include influenza viruses, parainfluenza viruses, cytomegaloviruses, adenoviruses, rhinoviruses, coronaviruses, and certain enteroviruses, such as Coxsackieviruses and echoviruses, as well as viruses such as herpes simplex, varicella-zoster virus, rubella, and measles. Infants and young children are also frequently infected with respiratory syncytial virus (RSV) and develop pneumonia. In addition, some enteroviruses, such as Coxsackieviruses and echoviruses, can also cause pneumonia symptoms through respiratory infection.
[0003] Pneumonia caused by various respiratory viral infections poses a serious threat to people's health, making it essential to find and discover new drugs to combat respiratory viruses.
[0004] Musk is the dried secretion of the prepuce of the male musk deer (Musk genus). It possesses various pharmacological activities and has been widely used in traditional Chinese medicine for thousands of years. Musk ketone is the main active ingredient of musk (structure shown below), and its pharmacological activities are similar to those of musk. It has been widely used to treat various diseases, such as nervous system disorders, chronic inflammation, and ischemia-reperfusion injury. However, the effects and mechanisms of musk or its active ingredients on respiratory viruses have not yet been reported.
[0005] Summary of the Invention
[0006] This invention provides the use of muscone in the preparation of antiviral drugs.
[0007] Furthermore, the aforementioned drugs are for the treatment and / or prevention of respiratory infections caused by respiratory viruses, wherein the respiratory infections are upper respiratory tract infections and / or lower respiratory tract infections.
[0008] Furthermore, the aforementioned medications for treating and / or preventing upper respiratory tract infections include medications for treating and / or preventing rhinitis, pharyngitis, sinusitis, otitis media, laryngitis, and / or epiglottitis.
[0009] The medications for treating and / or preventing lower respiratory tract infections include medications for treating and / or preventing tracheitis, bronchitis, and / or pneumonia.
[0010] Furthermore, the aforementioned drugs are those that reduce the activity of respiratory viruses.
[0011] Furthermore, the aforementioned respiratory viruses are coronaviruses, human metapneumoviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, measles viruses, mumps viruses, rubella viruses, adenoviruses, cytomegaloviruses, enteroviruses, herpes simplex viruses, varicella-zoster viruses, or rhinoviruses; the enteroviruses include Coxsackieviruses and echoviruses, preferably human metapneumoviruses.
[0012] The present invention also provides an antiviral drug, which is a preparation made of muscone as the active ingredient and pharmaceutically acceptable excipients or auxiliary ingredients.
[0013] Furthermore, the aforementioned respiratory viruses are coronaviruses, human metapneumoviruses, influenza viruses, parainfluenza viruses, respiratory syncytial viruses, measles viruses, mumps viruses, rubella viruses, adenoviruses, cytomegaloviruses, enteroviruses, herpes simplex viruses, varicella-zoster viruses, or rhinoviruses; the enteroviruses include Coxsackieviruses, echoviruses, and preferably human metapneumoviruses.
[0014] Furthermore, the content of the active ingredient in the above formulation is 1-1000 μM, preferably 6.25-880 μM, and more preferably 110-880 μM.
[0015] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.
[0016] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description
[0017] Figure 1 The results are the cell activity test results of muscone against 16HBE.
[0018] Figure 2 The results show the titer of HMPV virus in cells as determined by muscone.
[0019] Figure 3 The results show the HMPV virus titer test results of muscone on infected cells. Detailed Implementation
[0020] Unless otherwise stated, all raw materials used in this invention are known products, obtained by purchasing commercially available products.
[0021] This invention utilizes Vero-E6 cells to culture live human metapneumovirus (HMPV) virus, and isolates viral fluid (MOI=10) for subsequent experiments.
[0022] Example 1: Preparation of the drug containing muscone according to the present invention
[0023] Musk ketone was added to cell culture medium to prepare solutions with concentrations of 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, 200 μM, 27.5 μM, 55 μM, 110 μM, 220 μM, 440 μM, and 880 μM.
[0024] Experiment Example 1: Cytotoxicity Experiment
[0025] Cell resuscitation: Remove the frozen 16HBE cells from the liquid nitrogen tank and quickly place them in a 37°C water bath to thaw. Add the thawed cells to a 15ml BD tube containing 3ml of culture medium, centrifuge at 800rpm for 4min, discard the supernatant, add 1ml of culture medium to resuspend the cells, and then add the resuspended cells to a T25 culture flask and place it in an incubator for culture.
[0026] Cell plating: When 16HBE cells are cultured to approximately 80% confluence, they are digested, resuspended, and counted. The cell concentration is then diluted to 1*102. 5 / mL; 100μL was added to each well of a 96-well plate; cells were treated with different concentrations of muscone solution (0μM, 27.5μM, 55μM, 110μM, 220μM, 440μM, 880μM); control wells and blank wells were set up; cells were incubated overnight for 24h. The drug-containing culture medium was aspirated, the cells were washed three times with PBS, fresh culture medium was added, and 10μL of CCK-8 reagent was added; after 1-4h of incubation, the absorbance at 450nm was measured using a microplate reader, and the cell viability was calculated according to the following formula.
[0027] A = A C -A0 / A b -A0
[0028] A c For adding medicine;
[0029] A b Untreated wells containing cells;
[0030] A0 represents a cell-free, untreated well.
[0031] The results obtained are as follows Figure 1 As shown, the results indicate that the cell viability of 16HBE cells was above 80% under the action of various concentrations of muscone, indicating that muscone has no obvious cytotoxicity.
[0032] Experiment Example 2: Experiment to Inhibit Viral Activity
[0033] 1. The day before the experiment, after digesting and resuspending cells in the logarithmic growth phase, 2*10 5 HBE cells were seeded in 24-well plates and incubated overnight. On the second day, when cell confluence reached 70-80%, 200 μL of HMPV virus solution (MOI = 10) was added, and the cells were incubated for 2 hours. Virus was then aspirated and washed three times with PBS. Drug-containing solutions (0 μM, 6.25 μM, 12.5 μM, 25 μM, 50 μM, 100 μM, and 200 μM muscone solutions) were prepared, and the corresponding solutions were added to each well of the experimental groups. After 24 hours of incubation, the supernatant was discarded, and RNA was extracted using a total RNA extraction kit following the manufacturer's instructions. The extracted RNA was reverse transcribed using a reverse transcription kit (AG11706-S). Quantitative real-time PCR (Q-PCR) was performed using TaqMan fluorescent probes to investigate the intracellular viral load after adding different concentrations of muscone, based on the Cq values of different groups in the Q-PCR experiment.
[0034] The results are as follows Figure 2 As shown in the experimental results, the drug-containing solutions containing muscone can effectively reduce the intracellular viral titer. 6.25 μM muscone can significantly reduce the intracellular viral titer, indicating that muscone can effectively inhibit viral activity in cells.
[0035] 2. The day before the experiment, after digesting and resuspending cells in the logarithmic growth phase, 2*10 5 HBE cells were seeded in 24-well plates at 16 wells and grown overnight. The next day, thymol was added to the culture medium at corresponding concentrations (0 μM, 27.5 μM, 55 μM, 110 μM, 220 μM, 440 μM, 880 μM) and incubated for 24 h. Then, 200 μL of HMPV virus solution (MOI = 10) was added, and the culture was incubated at 37°C for 2 h. Following this, DMEM virus maintenance medium containing 3% FBS was added. 24 h after infection, RNA was extracted using a total RNA extraction kit, following the manufacturer's instructions. Reverse transcription and quantitative real-time PCR experiments were performed as described above.
[0036] The results are as follows Figure 3 As shown, the experimental results indicate that the titer of virus-infected cells decreased under the action of the drug. When the drug concentration reached above 110 μM, there was a significant difference compared with the control group, indicating that muscone can effectively inhibit viral activity in cells and has the potential to prevent HMPV virus infection.
[0037] In summary, this invention provides the use of muscone in the preparation of medicaments for the treatment and / or prevention of respiratory viral pneumonia. Medications with muscone as the active ingredient can effectively inhibit and / or treat respiratory viral infections such as coronavirus, human metapneumovirus, influenza virus, parainfluenza virus, respiratory syncytial virus, measles virus, mumps virus, rubella virus, adenovirus, cytomegalovirus, Coxsackie virus, echovirus, herpes simplex virus, varicella-zoster virus, or rhinovirus, and have promising clinical application prospects.
Claims
1. The use of muscone in the preparation of drugs against human metapneumovirus.
2. Use according to claim 1, characterized in that, The drug is a treatment and / or prevention drug for respiratory infections caused by human metapneumovirus, which are upper respiratory tract infections and / or lower respiratory tract infections.
3. The use as described in claim 2, characterized in that, The medications for treating and / or preventing upper respiratory tract infections include medications for treating and / or preventing rhinitis, pharyngitis, sinusitis, otitis media, laryngitis, and / or epiglottitis; the medications for treating and / or preventing lower respiratory tract infections include medications for treating and / or preventing tracheitis, bronchitis, and / or pneumonia.
4. The use as described in any one of claims 1 to 3, characterized in that, The drug in question is one that reduces the activity of human metapneumovirus.