A traditional Chinese medicine effective part composition, a preparation method and application thereof

Abstract

The application provides a traditional Chinese medicine effective part composition, a preparation method and application thereof. The traditional Chinese medicine effective part composition comprises the following weight parts of effective part medicines of traditional Chinese medicine compound preparation medicines: 125-2000 parts of polysaccharide parts, 0.625-2.5 parts of alkaloid parts, 6.25-100 parts of volatile oil parts and 6.25-100 parts of total glycoside parts. The traditional Chinese medicine compound preparation medicines comprise Poria cocos, Glycyrrhiza uralensis Fisch., Citrus aurantium, Platycodon grandiflorum, Bupleurum smithii, Gypsum fibrosum, Anemarrhena asphodeloides, Peucedanum decursivum, Notopterygium incisum, Cimicifuga foetida, Saposhnikovia divaricata, Schizonepeta, Chuanxiong, Pueraria lobata, Mentha haplocalyx, Lonicera japonica, Forsythia suspensa, Agastache rugosa, Ephedra sinica and Atractylodes lancea. The application further discloses a preparation method of the traditional Chinese medicine effective part composition and application of the traditional Chinese medicine effective part composition in preparing medicines for treating influenza and viral pneumonia. The influenza is influenza A, and the viral pneumonia is pneumonia induced by influenza A virus. A medicine for treating influenza and viral pneumonia comprises the traditional Chinese medicine effective part composition.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical technology, and in particular to a composition of effective components of traditional Chinese medicine, its preparation method, and its application. Background Technology

[0002] Influenza (hereinafter referred to as flu) is an acute respiratory infectious disease caused by the influenza virus. It is characterized by high infectivity, high morbidity, wide prevalence, and high mortality, and is considered one of the world's most prevalent infectious diseases that are currently not effectively controlled. my country is a country prone to influenza outbreaks, with outbreaks occurring almost every year, causing numerous critically ill patients and deaths, significantly impacting human health, life, education, and the economy. Influenza viruses are classified into three types: A, B, and C. Type A viruses frequently undergo antigenic mutations, are highly contagious, spread rapidly, and are prone to large-scale epidemics, making them the dominant type of influenza virus. Influenza often causes viral pneumonia, resulting from the downward spread of viral infection. Vaccines and antiviral drugs are effective means of preventing and treating influenza, but vaccines often face the possibility of becoming ineffective due to viral mutations. The first-line antiviral drug for treating influenza is oseltamivir phosphate capsules, which also faces limitations due to drug resistance caused by viral mutations and drug toxicity. Therefore, there is a clinical need for a safe, effective, low-toxicity, and inexpensive drug for the prevention and treatment of influenza.

[0003] Traditional Chinese medicine (TCM) considers influenza to fall under the category of exogenous febrile diseases, primarily affecting the lungs, but potentially extending to the stomach and intestines. The cause is exogenous febrile pathogens. Initially, the disease centers on the lungs and defensive qi (defensive qi). As the disease progresses, these pathogens penetrate deeper, affecting the lungs, defensive qi, and stomach and intestines. From a TCM perspective, the lungs and large intestine are internally and externally related, not only physiologically but also pathologically. Lung disease can affect the intestines, or vice versa; lung and intestinal diseases can spread and even create a vicious cycle. In the early stages of influenza, pathogens accumulate in the lungs and stomach, causing dampness and stagnation, impairing the lung's function of dispersing and descending qi, resulting in fever, runny nose, nasal congestion, cough, and sore throat. If the stomach and intestines are affected, disharmony occurs, disrupting the downward flow of qi, leading to nausea, vomiting, abdominal pain, diarrhea, and watery stools. Further progression results in the pathogens penetrating deeper, obstructing lung qi, damaging the lung vessels, and causing symptoms such as high fever, shortness of breath, rapid breathing, and constipation—symptoms of viral pneumonia.

[0004] The authorized patent "A Traditional Chinese Medicine Compound Preparation and Its Preparation Method and Application" (ZL2019111118087.1) is a traditional Chinese medicine compound preparation invented to address the shortcomings and deficiencies of the existing technology. This preparation, taken orally, can treat influenza and viral pneumonia. However, it suffers from drawbacks such as unclear pharmacokinetic mechanisms and unstable efficacy. Summary of the Invention

[0005] This application provides a composition of effective components of traditional Chinese medicine, its preparation method, and its application, to solve the problems existing in related technologies. The technical solution is as follows:

[0006] Based on the already authorized patented traditional Chinese medicine compound preparation, this application uses a systematic solvent extraction method to prepare the effective parts of the traditional Chinese medicine compound preparation, and conducts research on the efficacy of the formulation of the effective parts. The invention is a composition of the effective parts of traditional Chinese medicine. Oral administration of this composition of the effective parts of traditional Chinese medicine can achieve better therapeutic effects on influenza and viral pneumonia than the decoction of the original authorized patent.

[0007] This application provides a composition of effective components of traditional Chinese medicine. Based on the traditional Chinese medicine compound preparation in the authorized patent "A Traditional Chinese Medicine Compound Preparation and Its Preparation Method and Application" (application number 201911118087.1), the traditional Chinese medicine compound preparation is regarded as a whole. The system solvent method is used to extract and prepare the traditional Chinese medicine compound preparation into different chemical components. The in vitro and in vivo effects against influenza A virus are used as indicators to conduct pharmacological effects and compatibility formulation on the effective components of the original patent, thereby obtaining an optimized formula of highly efficient effective component combination.

[0008] In a first aspect, embodiments of this application provide a composition of effective components of a traditional Chinese medicine, comprising the following effective components of a compound traditional Chinese medicine preparation in parts by weight: 125-2000 parts of polysaccharide fraction, 0.625-2.5 parts of alkaloid fraction, 6.25-100 parts of volatile oil fraction, and 6.25-100 parts of total glycoside fraction.

[0009] The traditional Chinese medicine compound preparation includes: Poria cocos, licorice, immature bitter orange, platycodon root, bupleurum root, gypsum, anemarrhena rhizome, angelica root, notopterygium root, cimicifuga rhizome, saposhnikovia root, schizonepeta root, chuanxiong rhizome, kudzu root, peppermint root, honeysuckle flower, forsythia fruit, agastache rhizome, ephedra root, and atractylodes rhizome.

[0010] In one embodiment, the traditional Chinese medicine compound preparation comprises: 5-20 parts of Poria cocos, 5-10 parts of Glycyrrhiza uralensis, 5-20 parts of Citrus aurantium, 5-20 parts of Platycodon grandiflorus, 10-20 parts of Bupleurum chinense, 15-50 parts of Gypsum fibrosum, 10-20 parts of Anemarrhena asphodeloides, 10-20 parts of Peucedanum praeruptorum, 10-20 parts of Notopterygium incisum, 10-20 parts of Cimicifuga foetida, 10-20 parts of Saposhnikovia divaricata, 10-20 parts of Schizonepeta tenuifolia, 10-20 parts of Ligusticum chuanxiong, 10-30 parts of Pueraria lobata, 5-10 parts of Mentha haplocalyx, 15-30 parts of Lonicera japonica, 15-30 parts of Forsythia suspensa, 10-20 parts of Pogostemon cablin, 5-15 parts of Ephedra sinica, and 10-30 parts of Atractylodes lancea.

[0011] In one embodiment, the effective components of the traditional Chinese medicine compound preparation include the following parts by weight: 2000 parts of polysaccharide fraction, 0.625 parts of alkaloid fraction, 6.25 parts of volatile oil fraction, and 6.25 parts of total glycoside fraction.

[0012] The traditional Chinese medicine compound preparation includes: 10 parts Poria cocos, 5 parts Glycyrrhiza uralensis, 10 parts Citrus aurantium, 10 parts Platycodon grandiflorus, 10 parts Bupleurum chinense, 30 parts Gypsum fibrosum, 10 parts Anemarrhena asphodeloides, 10 parts Peucedanum praeruptorum, 10 parts Notopterygium incisum, 10 parts Cimicifuga foetida, 10 parts Saposhnikovia divaricata, 10 parts Schizonepeta tenuifolia, 10 parts Ligusticum chuanxiong, 15 parts Pueraria lobata, 5 parts Mentha haplocalyx, 15 parts Lonicera japonica, 15 parts Forsythia suspensa, 15 parts Pogostemon cablin, 10 parts Ephedra sinica, and 15 parts Atractylodes lancea.

[0013] In one embodiment, the extraction method of the alkaloid fraction includes: weighing the traditional Chinese medicine compound preparation in proportion, refluxing with 70% ethanol for extraction, concentrating under reduced pressure until there is no alcohol odor, and obtaining a concentrated ethanol extract, with the residue reserved for later use.

[0014] Add a 5% NaOH solution to the concentrated ethanol extract to adjust the pH to 12, extract with chloroform, separate the liquids, take the chloroform extract, concentrate under reduced pressure, and dry to obtain the chloroform extract, which is the alkaloid fraction.

[0015] The extraction method of the polysaccharide fraction includes: reflux extraction of the medicinal residue with water, filtration of the extract, concentration to the amount of the medicinal material, addition of 95% ethanol, mixing, overnight standing, filtration or centrifugation, collection of the precipitate, freeze-drying or vacuum drying at 60-80℃ to obtain the polysaccharide fraction.

[0016] In one embodiment, the method for extracting the total glycoside fraction includes:

[0017] Weigh out the combination of Chinese herbs, reflux extract with 70% ethanol, concentrate under reduced pressure until there is no alcohol taste, and obtain ethanol extract concentrate; add n-butanol for extraction, separate the liquids, take the n-butanol extract, concentrate under reduced pressure, and dry under vacuum to obtain the n-butanol extract fraction, which is the total glycoside fraction;

[0018] The extraction method of the volatile oil fraction includes: weighing the combination of Chinese medicines, placing them in a steam distillation jar, adding 12 times the amount of water to soak them, distilling and extracting them, and collecting the volatile oil fraction.

[0019] Secondly, embodiments of this application provide a method for preparing a composition of effective components of traditional Chinese medicine, used to prepare the composition of effective components of traditional Chinese medicine as described in any of the above claims, wherein the composition of effective components of traditional Chinese medicine is a tablet, and the preparation method includes:

[0020] The polysaccharide fraction, alkaloid fraction, volatile oil fraction, and total glycoside fraction are mixed evenly in a certain proportion, spray-dried, and then dextrin and sucrose powder are added in a weight ratio of effective fraction mixture: dextrin: sucrose powder = 1:1:3. The mixture is mixed evenly, passed through a 60-mesh sieve to obtain granules, dried, and compressed into tablets.

[0021] Thirdly, embodiments of this application provide a method for preparing a composition of effective components of traditional Chinese medicine, used to prepare the composition of effective components of traditional Chinese medicine as described in any of the above claims, wherein the composition of effective components of traditional Chinese medicine is in the form of granules, capsules, or pills, and the preparation method includes:

[0022] Weigh out the traditional Chinese medicine compound preparations according to the specified proportions, mix the alkaloid fraction, polysaccharide fraction, total glycoside fraction, and volatile oil fraction, and add excipients to prepare granules, capsules, or pills. The excipients used are common pharmaceutical excipients and will not be described in detail here.

[0023] Fourthly, embodiments of this application provide the application of a composition of effective components of traditional Chinese medicine in the preparation of drugs for treating influenza and viral pneumonia.

[0024] In one embodiment, the influenza is influenza A, and the viral pneumonia is pneumonia induced by influenza A virus.

[0025] Fifthly, embodiments of this application provide a medicament for treating influenza and viral pneumonia, comprising a composition of effective components of traditional Chinese medicine as described in any of the above-mentioned embodiments.

[0026] The advantages or beneficial effects of the above technical solutions include at least the following:

[0027] Based on the viral pneumonia model test of influenza A virus infection, this application has confirmed that the effective component composition of traditional Chinese medicine has a more significant effect on increasing the survival rate of virus-infected mice, reducing inflammatory damage to the lungs, and inhibiting the level of inflammatory cytokines in lung tissue than the decoction (concentrated decoction) of the original patented traditional Chinese medicine compound preparation. It can significantly treat acute lung injury induced by influenza virus.

[0028] The composition of effective components of traditional Chinese medicine can be used to treat influenza and its viral pneumonia. In this application, influenza includes influenza A; viral pneumonia includes, but is not limited to, pneumonia induced by influenza A virus, and can also be used for viral pneumonia induced by other viruses.

[0029] The above overview is for illustrative purposes only and is not intended to be limiting in any way. In addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features of this application will become readily apparent from the accompanying drawings and the following detailed description. Attached Figure Description

[0030] In the accompanying drawings, unless otherwise specified, the same reference numerals throughout the various drawings denote the same or similar parts or elements. These drawings are not necessarily drawn to scale. It should be understood that these drawings depict only some embodiments disclosed in this application and should not be construed as limiting the scope of this application.

[0031] Figure 1 The figure shows the experimental results of the effect of influenza A virus infection on the lung index of mice.

[0032] Figure 2 Figure showing the experimental results of the effect of influenza A virus infection on the lung pathology and morphology of mice.

[0033] Figure 3 The figure shows the experimental results of the effect of influenza A virus on the titer of mouse pneumovirus.

[0034] Figure 4 -8 shows the experimental results of the effect of influenza A virus infection on mouse lung cytokines.

[0035] Figure 9 -10 shows the experimental results of the protective effect against influenza A virus infection in mice. Detailed Implementation

[0036] To better illustrate the purpose, technical solution, and advantages of this application, the following embodiments are provided.

[0037] In the following description, only certain exemplary embodiments are briefly described. As those skilled in the art will recognize, the described embodiments can be modified in various ways without departing from the spirit or scope of this application. Therefore, the drawings and description are considered to be exemplary in nature and not restrictive.

[0038] In a first aspect, embodiments of this application provide a composition of effective components of traditional Chinese medicine, comprising the following effective components of a traditional Chinese medicine compound preparation in parts by weight: 125-2000 parts of polysaccharide fraction, 0.625-2.5 parts of alkaloid fraction, 6.25-100 parts of volatile oil fraction, and 6.25-100 parts of total glycoside fraction.

[0039] Among them, the traditional Chinese medicine compound preparations include: Poria cocos, licorice, immature bitter orange, platycodon root, bupleurum root, gypsum, anemarrhena rhizome, angelica root, notopterygium root, cimicifuga rhizome, saposhnikovia root, schizonepeta root, chuanxiong rhizome, kudzu root, peppermint root, honeysuckle flower, forsythia fruit, agastache rhizome, ephedra root, and atractylodes rhizome.

[0040] In one embodiment, the traditional Chinese medicine compound preparation includes: 5-20 parts of Poria cocos, 5-10 parts of Glycyrrhiza uralensis, 5-20 parts of Citrus aurantium, 5-20 parts of Platycodon grandiflorus, 10-20 parts of Bupleurum chinense, 15-50 parts of Gypsum fibrosum, 10-20 parts of Anemarrhena asphodeloides, 10-20 parts of Peucedanum praeruptorum, 10-20 parts of Notopterygium incisum, 10-20 parts of Cimicifuga foetida, 10-20 parts of Saposhnikovia divaricata, 10-20 parts of Schizonepeta tenuifolia, 10-20 parts of Ligusticum chuanxiong, 10-30 parts of Pueraria lobata, 5-10 parts of Mentha haplocalyx, 15-30 parts of Lonicera japonica, 15-30 parts of Forsythia suspensa, 10-20 parts of Pogostemon cablin, 5-15 parts of Ephedra sinica, and 10-30 parts of Atractylodes lancea.

[0041] In one embodiment, the effective component composition of traditional Chinese medicine comprises the following effective components of the traditional Chinese medicine compound preparation in parts by weight: 2000 parts of polysaccharide component, 0.625 parts of alkaloid component, 6.25 parts of volatile oil component, and 6.25 parts of total glycoside component.

[0042] The herbal combination includes: 10 parts Poria cocos, 5 parts Glycyrrhiza uralensis, 10 parts Citrus aurantium, 10 parts Platycodon grandiflorus, 10 parts Bupleurum chinense, 30 parts Gypsum fibrosum, 10 parts Anemarrhena asphodeloides, 10 parts Peucedanum praeruptorum, 10 parts Notopterygium incisum, 10 parts Cimicifuga foetida, 10 parts Saposhnikovia divaricata, 10 parts Schizonepeta tenuifolia, 10 parts Ligusticum chuanxiong, 15 parts Pueraria lobata, 5 parts Mentha haplocalyx, 15 parts Lonicera japonica, 15 parts Forsythia suspensa, 15 parts Pogostemon cablin, 10 parts Ephedra sinica, and 15 parts Atractylodes lancea.

[0043] In one embodiment, the method for extracting alkaloids includes: weighing a combination of traditional Chinese medicines, refluxing with 70% ethanol, concentrating under reduced pressure until no alcohol odor remains, obtaining a concentrated ethanol extract, and reserving the residue.

[0044] Add a 5% NaOH solution to the concentrated ethanol extract to adjust the pH to 12, extract with chloroform, separate the liquids, concentrate the chloroform extract under reduced pressure, and dry to obtain the chloroform extract, which is the alkaloid fraction.

[0045] In one embodiment, the extract was refluxed with 70% ethanol three times, each time for 1.5 hours. The ethanol extracts were combined and concentrated under reduced pressure until no alcohol odor remained.

[0046] Extracted three times with chloroform.

[0047] In one embodiment, the method for extracting the polysaccharide fraction includes: refluxing the medicinal residue with water for extraction, filtering the extract, concentrating it to the amount of the medicinal material, adding 95% ethanol, mixing well, letting it stand overnight, filtering or centrifuging, taking the precipitate, freeze-drying or vacuum drying at 60-80°C to obtain the polysaccharide fraction.

[0048] The final total polysaccharide yield (each gram of extract equivalent to grams of raw medicinal herbs) was 6.148%.

[0049] In one embodiment, the method for extracting total glycosides includes:

[0050] Weigh out the combination of Chinese herbs, reflux extract with 70% ethanol, concentrate under reduced pressure until there is no alcohol odor, and obtain ethanol extract concentrate; add n-butanol for extraction, separate the liquids, take the n-butanol extract, concentrate under reduced pressure, and dry under vacuum to obtain the n-butanol extract fraction, which is the total glycoside fraction.

[0051] In one embodiment, the extract was refluxed with 70% ethanol three times, each time for 1.5 hours. The ethanol extracts were combined and concentrated under reduced pressure until no alcohol odor remained.

[0052] Extract three times with n-butanol;

[0053] The vacuum drying temperature is 60℃.

[0054] In one embodiment, the method for extracting the volatile oil fraction includes: weighing a combination of Chinese herbal medicines, placing them in a steam distillation jar, adding 12 times the amount of water to soak them, distilling and extracting them, and collecting the volatile oil fraction.

[0055] In one embodiment, the sample is soaked in 12 times the amount of water for 2 hours and then extracted by distillation for 10 hours.

[0056] Secondly, embodiments of this application provide a method for preparing a composition of effective components of traditional Chinese medicine, used to prepare the composition of effective components of traditional Chinese medicine as described in any of the above claims, wherein the composition of effective components of traditional Chinese medicine is a tablet, and the preparation method includes:

[0057] The polysaccharide fraction, alkaloid fraction, volatile oil fraction, and total glycoside fraction are mixed evenly in a certain proportion, spray-dried, and then dextrin and sucrose powder are added in a weight ratio of effective fraction mixture: dextrin: sucrose powder = 1:1:3. The mixture is mixed evenly, passed through a 60-mesh sieve to obtain granules, dried, and compressed into tablets.

[0058] Thirdly, embodiments of this application provide a method for preparing a composition of effective components of traditional Chinese medicine, used to prepare any of the above-mentioned compositions of effective components of traditional Chinese medicine, wherein the composition of effective components of traditional Chinese medicine is in the form of granules, capsules, or pills, and the preparation method includes:

[0059] Weigh out the traditional Chinese medicine compound preparations according to the specified proportions, mix the alkaloid fraction, polysaccharide fraction, total glycoside fraction, and volatile oil fraction, and add excipients to prepare granules, capsules, or pills. The excipients used are common pharmaceutical excipients and will not be described in detail here.

[0060] Fourthly, embodiments of this application provide the application of a composition of effective components of traditional Chinese medicine in the preparation of drugs for treating influenza and viral pneumonia.

[0061] In one implementation, influenza refers to influenza A, and viral pneumonia refers to pneumonia induced by influenza A virus.

[0062] Fifthly, embodiments of this application provide a medicament for treating influenza and viral pneumonia, comprising a composition of effective components of traditional Chinese medicine.

[0063] Example 1

[0064] This embodiment provides a composition of effective components of traditional Chinese medicine, which is a tablet comprising the following effective components of a traditional Chinese medicine compound preparation in parts by weight: 2000 parts of polysaccharide fraction, 0.625 parts of alkaloid fraction, 6.25 parts of volatile oil fraction, and 6.25 parts of total glycoside fraction.

[0065] The traditional Chinese medicine compound preparation includes: 10 parts Poria cocos, 5 parts Glycyrrhiza uralensis, 10 parts Citrus aurantium, 10 parts Platycodon grandiflorus, 10 parts Bupleurum chinense, 30 parts Gypsum fibrosum, 10 parts Anemarrhena asphodeloides, 10 parts Peucedanum praeruptorum, 10 parts Notopterygium incisum, 10 parts Cimicifuga foetida, 10 parts Saposhnikovia divaricata, 10 parts Schizonepeta tenuifolia, 10 parts Ligusticum chuanxiong, 15 parts Pueraria lobata, 5 parts Mentha haplocalyx, 15 parts Lonicera japonica, 15 parts Forsythia suspensa, 15 parts Pogostemon cablin, 10 parts Ephedra sinica, and 15 parts Atractylodes lancea.

[0066] The extraction method of alkaloids includes: weighing 2 kg of traditional Chinese medicine compound preparation, refluxing with 70% ethanol three times, each time for 1.5 h, combining the ethanol extracts, concentrating under reduced pressure until there is no alcohol taste, and obtaining a concentrated ethanol extract, with the residue reserved.

[0067] Add a 5% NaOH solution to the concentrated ethanol extract to adjust the pH to 12, extract three times with chloroform, separate the liquids, collect the chloroform extract, concentrate under reduced pressure, and dry to obtain the chloroform extract, which is the alkaloid fraction.

[0068] Ultimately, the yield of the alkaloid fraction (each gram of extract equivalent to grams of raw medicinal slices) was 1.174%.

[0069] The extraction method of polysaccharide fraction includes: adding water to the residue for reflux extraction, filtering the extract, concentrating it to the amount of the medicinal material, adding 95% ethanol, mixing well, letting it stand overnight, filtering or centrifuging, taking the precipitate, freeze-drying or vacuum drying at 60-80℃ to obtain the polysaccharide fraction.

[0070] Ultimately, the total polysaccharide yield (each gram of extract equivalent to grams of raw medicinal herbs) was 6.148%.

[0071] Methods for extracting total glycosides include:

[0072] Weigh 2 kg of the traditional Chinese medicine compound preparation and extract it three times with 70% ethanol for 1.5 h each time. Combine the ethanol extracts and concentrate them under reduced pressure until there is no alcohol taste to obtain the concentrated ethanol extract. Add n-butanol and extract three times. Separate the liquids and take the n-butanol extract. Concentrate it under reduced pressure and dry it under vacuum at 60℃ to obtain the n-butanol extract, which is the total glycoside fraction.

[0073] Ultimately, the total glycoside yield (each gram of extract equivalent to grams of raw medicinal herbs) was 11.042%.

[0074] The extraction method of the volatile oil fraction includes: weighing 4 kg of the Chinese herbal medicine combination, placing it in a steam distillation jar, adding 12 times the amount of water to soak for 2 hours, distilling and extracting for 10 hours, and collecting the volatile oil fraction.

[0075] Ultimately, the yield of volatile oil components (each gram of extract equivalent to the number of grams of raw medicinal herbs) was 0.25%.

[0076] The above-mentioned effective components are mixed evenly in proportion, spray-dried, and then dextrin and sucrose powder are added in a weight ratio of effective component mixture: dextrin: sucrose powder = 1:1:3. The mixture is mixed evenly, passed through a 60-mesh sieve to obtain granules, dried, and compressed into tablets, each weighing 0.5g.

[0077] The dosage of this tablet, which contains the effective components of traditional Chinese medicine, is 3 tablets three times a day.

[0078] Example 2

[0079] This embodiment provides a composition of effective components of traditional Chinese medicine. This embodiment is a soft capsule, comprising the following effective components of a traditional Chinese medicine compound preparation in parts by weight: 2000 parts of polysaccharide fraction, 0.625 parts of alkaloid fraction, 6.25 parts of volatile oil fraction, and 6.25 parts of total glycoside fraction.

[0080] The traditional Chinese medicine compound preparation includes: 10 parts Poria cocos, 5 parts Glycyrrhiza uralensis, 10 parts Citrus aurantium, 10 parts Platycodon grandiflorus, 10 parts Bupleurum chinense, 30 parts Gypsum fibrosum, 10 parts Anemarrhena asphodeloides, 10 parts Peucedanum praeruptorum, 10 parts Notopterygium incisum, 10 parts Cimicifuga foetida, 10 parts Saposhnikovia divaricata, 10 parts Schizonepeta tenuifolia, 10 parts Ligusticum chuanxiong, 15 parts Pueraria lobata, 5 parts Mentha haplocalyx, 15 parts Lonicera japonica, 15 parts Forsythia suspensa, 15 parts Pogostemon cablin, 10 parts Ephedra sinica, and 15 parts Atractylodes lancea.

[0081] The preparation method of this effective component composition of traditional Chinese medicine is as follows:

[0082] The extraction method for alkaloids includes: weighing 2 kg of the traditional Chinese medicine compound preparation according to the specified proportion, extracting it three times with 70% ethanol under reflux for 1.5 hours each time, combining the ethanol extracts, concentrating under reduced pressure until no alcohol odor remains, and obtaining a concentrated ethanol extract. The residue is then set aside. A 5% NaOH solution is added to the concentrated ethanol extract to adjust the pH to 12, and the mixture is extracted three times with chloroform. The extracts are separated, concentrated under reduced pressure, and dried to obtain the chloroform extract, which is the alkaloid fraction. Ultimately, the yield of the alkaloid fraction (each gram of extract equivalent to grams of raw medicinal slices) is 1.174%.

[0083] The extraction method for total polysaccharides includes: reflux extraction of the residue with water, filtration of the extract, concentration to the amount of raw medicinal material, addition of 95% ethanol, mixing, overnight standing, filtration or centrifugation, collection of the precipitate, freeze-drying or vacuum drying at 60-80℃ to obtain the total polysaccharide fraction. Ultimately, the yield of total polysaccharides (each gram of extract equivalent to grams of raw medicinal material) was 6.148%.

[0084] The extraction method for total glycosides includes: weighing 2 kg of the traditional Chinese medicine compound preparation according to the specified ratio, extracting it three times with 70% ethanol for 1.5 hours each time, combining the ethanol extracts, concentrating under reduced pressure until no alcohol odor remains, and obtaining a concentrated ethanol extract; extracting three times with n-butanol, separating the liquids, taking the n-butanol extract, concentrating under reduced pressure, and drying under vacuum at 60℃ to obtain the n-butanol extract, which is the glycoside fraction. Ultimately, the yield of total glycosides (each gram of extract equivalent to grams of raw medicinal slices) was 11.042%.

[0085] The extraction method for the volatile oil fraction includes: weighing 4 kg of the traditional Chinese medicine compound preparation according to the specified ratio, placing it in a steam distillation vessel, adding 12 times the amount of water to soak for 2 hours, distilling and extracting for 10 hours, and collecting the volatile oil fraction. The final yield of the volatile oil fraction (each gram of the extract equivalent to the number of grams of raw medicinal material) is 0.25%.

[0086] The above-mentioned effective components are mixed evenly in proportion, spray-dried, and then an appropriate amount of soybean oil, soybean lecithin, beeswax and polyethylene glycol are added to prepare a suspension. Soft capsules are prepared by compression method, with each capsule containing 0.5g of drug.

[0087] The dosage of this soft capsule, containing the effective components of traditional Chinese medicine, is as follows: 2 capsules three times a day.

[0088] Example 3

[0089] This embodiment studies the formulation ratio of effective components of traditional Chinese medicine to combat influenza.

[0090] Using uniform design table U12 ＊ (12 10 Using in vitro cell culture methods, different chemical fractions isolated were combined to create anti-influenza virus combinations. A549 cells were infected with H1N1 influenza A virus (1 / 47 / FM strain), and cells were treated with different concentrations of the drug for in vitro screening. First, the drug's cytotoxicity to MDCK cells (TC50) and viral titer (TCID) were measured. 50 Based on the results of the cytotoxicity assay, further experiments were conducted within the maximum non-toxic concentration range. The viral titer was controlled at 100 TCID. 50 The drug was administered via a specific route of administration. Cell morphology changes were observed daily under an inverted microscope, and cytopathic effect (CPE) was recorded. The MTT assay was used for colorimetric analysis to calculate the drug's viral inhibition rate (%). Analysis was performed using uniform design software 3.0 to determine the optimal combination of effective components of the traditional Chinese medicine for anti-influenza treatment. The results are shown in Table 1.

[0091] Table 1. Screening and efficacy of effective components of traditional Chinese medicine against influenza (μg / ml)

[0092]

[0093] Multiple regression analysis yielded the following regression equation: y = 19.0 + 1.76e⁻²*X(1) - 0.992)*X(2) - 5.91e⁻²*X(3) - 8.98e⁻³*X(4). The multiple correlation coefficient was R = 0.9233, the coefficient of determination was R² = 0.8525, and the adjusted coefficient of determination R²a = 0.7972. The regression equation was tested with a sample size of N = 12, a significance level of α = 0.05, a test value of Ft = 10.11, and a critical value of F(0.05, 4, 7) = 4.120. Since Ft > F(0.05, 4, 7), the regression equation was significant. The predicted optimal combination was: polysaccharide fraction: alkaloid fraction: volatile oil fraction: total glycoside fraction = 2000: 0.625: 6.25: 6.25. The predicted inhibition rate was 53.1%. The combination was verified to have an actual virus inhibition rate of 50.80%.

[0094] Therefore, the optimal ratio of the effective components of traditional Chinese medicine for anti-influenza was determined to be: polysaccharide fraction: alkaloid fraction: volatile oil fraction: total glycoside fraction = 2000: 0.625: 6.25: 6.25.

[0095] Example 4

[0096] This embodiment studies the effects of a combination of effective components of traditional Chinese medicine on influenza virus pneumonia in mice.

[0097] The optimal ratio of effective components of traditional Chinese medicine was compared with the decoction of a traditional Chinese medicine compound preparation (application number 201911118087.1) and the normal control, model control, and the model decoction.

[0098] Sixty BALB / c mice (13-15g) were randomly divided into 5 groups according to body weight, with 10 mice in each group: normal control group; influenza A virus control group; and control group (…). Figure 1 The virus control group; the ribavirin group (100 mg / kg); the decoction group of the traditional Chinese medicine compound preparation with application number 201911118087.1 ( Figure 1 The decoction group of the Chinese patent formula was prepared according to the prescription and method of Example 1 in the authorized patent, and concentrated into 200mL of Chinese herbal decoction, equivalent to 6g / kg of raw herb; the effective part composition group of Chinese herbal medicine in this application ( Figure 1 The effective component combination of the Chinese patent formula, 5g / kg, was prepared at a ratio of polysaccharide: alkaloid: volatile oil: glycoside = 2000: 0.625: 6.25: 6.25. Mice in the remaining groups (excluding the normal control group) were anesthetized with ether and infected nasally with 15 LD. 5050 μL of 1 / 47 / FM / H1N1 influenza virus solution was used, while 50 μL of physiological saline was administered intranasally as a control group. Two hours post-infection, mice were administered the drug via gavage. Both the normal and virus control groups received the same volume of physiological saline (40 mL / kg) once daily for four consecutive days. On the fifth day post-infection, the animals were sacrificed, and lung weight and body weight were recorded. Blood was collected from the abdominal aorta, and the entire lung was removed from the thoracic cavity. Lung lesions were visually observed, and the extent of lesions was recorded. The lungs were washed with physiological saline, and residual saline was absorbed with clean filter paper. The wet weight was measured, and the lung index was calculated. A portion of the mouse lung was fixed in 10% paraformaldehyde solution and processed according to standard procedures. HE staining and sectioning were performed. Pathological lesions in the lung tissue were observed, and a portion of the lung tissue was used to detect cytokines and viral titers. In this experiment, the drug was administered at a volume of 20 mL / kg of mouse body weight.

[0099] The results are as follows:

[0100] (1) Lung Index: On the fourth day after viral infection, the animals were weighed, and the lungs were weighed after routine treatment; this was taken as the wet weight of the lungs. The lung index was calculated as: Lung Index = [Wet weight of lungs (mg) / Body weight (g)] × 100. The experimental results are as follows: Figure 1 As shown, compared with the virus control group, both the decoction of the traditional Chinese medicine compound preparation with application number 201911118087.1 and the composition of effective parts of traditional Chinese medicine in this application significantly inhibited the increase of lung index in mice (P < 0.01). The composition of effective parts of traditional Chinese medicine in this application was superior to the decoction in inhibiting the lung index in mice, and the difference between the two was statistically significant (p < 0.05, **p < 0.05, **p < 0.01, ****p < 0.001). Compared with the decoction, #: p < 0.05; ##: p < 0.01.

[0101] (2) Lung tissue pathology: HE results are as follows Figure 2 As shown, the lung tissue of mice in the normal control group had normal structure and no inflammatory lesions. The lung tissue of mice in the virus control group showed varying degrees of inflammatory lesions, with extensive inflammatory cell infiltration in the alveolar walls, widening of the alveolar walls, unclear structure, and hemorrhage and edema. The decoction of a traditional Chinese medicine compound preparation with application number 201911118087.1 (…) Figure 2 Traditional Chinese medicine compound preparations (decoction) and the composition of effective parts of traditional Chinese medicine in this application ( Figure 2 The combination of effective components significantly improved pneumonia pathology and reduced lung pathology scores in influenza virus-infected mice. (Note: Compared with the virus control group, *p<0.05, **p<0.01, ***p<0.01).

[0102] (3) Viral load in the lungs: A 100 mg sample of right lung tissue from mice was taken, and the viral load in the lung tissue was determined by real-time quantitative RT-PCR. The experimental results are as follows: Figure 3As shown, compared with the virus control group, both the patented decoction (a compound traditional Chinese medicine preparation) and the combination of effective components significantly inhibited the increase of viral titer in the lungs of infected mice (P < 0.05, P < 0.01), indicating that both the patented decoction and the combination of effective components can effectively inhibit viral replication in mouse lungs. (Note: Compared with the virus control group, *p < 0.05, **p < 0.01, ***p < 0.01).

[0103] (4) Lung tissue cytokines: Mouse lung homogenate was centrifuged at 3000 rpm for 15 minutes to collect the supernatant. The levels of TNF-α, IL-4, IL-1β, IFN-γ, and MCP-1 were detected by ELISA. The results are as follows: Figure 4 As shown in Figure 8, compared with the virus control group, the combination of the patented decoction and the effective components significantly reduced the levels of pro-inflammatory factors TNF-α, IL-4, IL-1β, IFN-γ, and MCP-1 in mouse lungs (P < 0.05–0.001). This indicates that oral administration of the traditional Chinese medicine compound preparation of this invention has a significant inhibitory effect on H1N1 influenza virus-induced acute pneumonia in mice. (Note: Compared with the virus control group, *p < 0.05, **p < 0.01, ***p < 0.001).

[0104] Example 5

[0105] This embodiment presents an experimental study on the survival protection of mice infected with influenza virus.

[0106] Fifty BALB / c mice (13-15g) were used. The experimental groups, infection doses, and administration regimens were the same as in Experiment 3. Except for the normal control group, the other mice were anesthetized with ether and intranasally infected with 50μL of 5LD50 1 / 47 / FM / H1N1 influenza virus solution. The normal control group was controlled by intranasal administration of 50μL of physiological saline. Two hours after infection, the mice were administered the drug by gavage. The normal control group and the virus control group were administered the same volume of physiological saline (20mL / kg) by gavage. The drug was administered once a day for 6 consecutive days. After drug withdrawal, the mice were observed for 14 days. The number of surviving and dying animals was recorded daily, and the life protection rate of the drug in influenza-infected mice was calculated.

[0107] Experimental results confirmed that the mortality rate of the patented decoction was 100%, with a survival time of 4.5 ± 0.08 days, while the mortality rate of the patented effective component combination was 80%, with an average survival time of 8.4 ± 4.8 days. Compared with the virus control group and the decoction, the patented effective component combination prolonged the survival time of mice, demonstrating a certain degree of survival protection. Figure 9 , 10 Note: (A) Mouse survival rate; (B) Average survival time of mice. Compared with the virus control group, **p<0.01, **p<0.01). Compared with the decoction, #: p<0.05; ##: p<0.01.

[0108] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this application. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of those different embodiments or examples.

[0109] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of this application, "a plurality of" means two or more, unless otherwise explicitly specified.

[0110] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any person skilled in the art can easily conceive of various variations or substitutions within the technical scope disclosed in this invention, and these should all be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A composition of effective components of traditional Chinese medicine, characterized in that... This traditional Chinese medicine compound preparation is composed of the following effective parts by weight: 2000 parts polysaccharide, 0.625 parts alkaloid, 6.25 parts volatile oil, and 6.25 parts total glycosides. The traditional Chinese medicine compound preparation consists of: 10 parts Poria cocos, 5 parts Glycyrrhiza uralensis, 10 parts Citrus aurantium, 10 parts Platycodon grandiflorus, 10 parts Bupleurum chinense, 30 parts Gypsum fibrosum, 10 parts Anemarrhena asphodeloides, 10 parts Peucedanum praeruptorum, 10 parts Notopterygium incisum, 10 parts Cimicifuga foetida, 10 parts Saposhnikovia divaricata, 10 parts Schizonepeta tenuifolia, 10 parts Ligusticum chuanxiong, 15 parts Pueraria lobata, 5 parts Mentha haplocalyx, 15 parts Lonicera japonica, 15 parts Forsythia suspensa, 15 parts Pogostemon cablin, 10 parts Ephedra sinica, and 15 parts Atractylodes lancea. The extraction method of the alkaloid fraction includes: weighing the traditional Chinese medicine compound preparation in proportion, refluxing with 70% ethanol, concentrating under reduced pressure until there is no alcohol odor, obtaining a concentrated ethanol extract, and reserving the residue; adding a 5% NaOH solution to the concentrated ethanol extract, adjusting the pH to 12, extracting with chloroform, separating the liquids, taking the chloroform extract, concentrating under reduced pressure, drying, and obtaining the chloroform extract, which is the alkaloid fraction; The extraction method of the polysaccharide fraction includes: adding water to the residue for reflux extraction, filtering the extract, concentrating it to the amount of the medicinal material, adding 95% ethanol, mixing well, letting it stand overnight, filtering or centrifuging, taking the precipitate, freeze-drying or vacuum drying at 60-80℃ to obtain the polysaccharide fraction. The extraction method of the total glycoside fraction includes: weighing the combination of Chinese medicines, refluxing with 70% ethanol, concentrating under reduced pressure until there is no alcohol odor, and obtaining a concentrated ethanol extract; adding n-butanol for extraction, separating the liquids, taking the n-butanol extract, concentrating under reduced pressure, and drying under vacuum to obtain the n-butanol extract fraction, which is the total glycoside fraction. The extraction method of the volatile oil fraction includes: weighing the combination of Chinese medicines, placing them in a steam distillation jar, adding 12 times the amount of water to soak them, distilling and extracting them, and collecting the volatile oil fraction.

2. A method for preparing a composition of effective components of traditional Chinese medicine, used to prepare the composition of effective components of traditional Chinese medicine as described in claim 1, characterized in that, The composition of the effective components of the traditional Chinese medicine is in the form of tablets, and its preparation method includes: The polysaccharide fraction, alkaloid fraction, volatile oil fraction, and total glycoside fraction are mixed evenly in a certain proportion, spray-dried, and then dextrin and sucrose powder are added in a weight ratio of effective fraction mixture: dextrin: sucrose powder = 1:1:

3. The mixture is mixed evenly, passed through a 60-mesh sieve to obtain granules, dried, and compressed into tablets.

3. A method for preparing a composition of effective components of traditional Chinese medicine, used to prepare the composition of effective components of traditional Chinese medicine as described in claim 1, characterized in that, The composition of the effective components of the traditional Chinese medicine is in the form of granules, capsules, or pills, and its preparation method includes: Weigh out the traditional Chinese medicine compound preparations according to the specified proportions, mix the alkaloid fraction, polysaccharide fraction, total glycoside fraction and volatile oil fraction, add excipients to make granules, capsules or pills.

4. The use of the effective component composition of traditional Chinese medicine according to claim 1 in the preparation of drugs for treating influenza and viral pneumonia.

5. The application according to claim 4, characterized in that, The influenza is influenza A, and the viral pneumonia is pneumonia induced by influenza A virus.

6. A drug for treating influenza and its viral pneumonia, characterized in that, The composition includes the effective components of traditional Chinese medicine as described in claim 1.

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