Disclosure of Invention
The invention provides application of a traditional Chinese medicine composition in preparing a medicine for preventing and treating novel pneumonia caused by coronavirus infection, wherein the traditional Chinese medicine composition is prepared from the following raw material medicines in parts by weight:
30-55 parts of forsythia fruit 150-
Patchouli 50-90 rhizoma dryopteris crassirhizomae 150-260 rhodiola rosea 50-90 menthol 5-8
50-90 fried Chinese ephedra, 50-90 fried bitter apricot seed, 50-90 cordate houttuynia, 150-
Licorice root 50-90 gypsum 150-260.
The traditional Chinese medicine composition disclosed by the invention preferably comprises the following raw material medicines in parts by weight:
forsythia fruit 150 honeysuckle 260 isatis root 150 rhubarb 55 patchouli 50
Rhizoma dryopteris crassirhizomae 260 rhodiola rosea 50 menthol crystal and ephedra herb 50 roasted with 8
Parched semen Armeniacae amarum 90, herba Houttuyniae 150, Glycyrrhrizae radix 90 and Gypsum Fibrosum 150.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
forsythia fruit 260 honeysuckle 150 isatis root 260 rhubarb 30 patchouli 90
Rhizoma Dryopteris Crassirhizomatis 150 radix Rhodiolae 90 Mentholum 5 herba Ephedrae 90
Parched semen Armeniacae amarum 50 herba Houttuyniae 260 Glycyrrhrizae radix 50 Gypsum Fibrosum 260.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
forsythia fruit 170 honeysuckle 170 isatis root 170 rhubarb 34 patchouli 57
Rhizoma dryopteris crassirhizomae 170 rhodiola rosea 57 menthol crystal 5 mix-fried ephedra 57
Parched semen Armeniacae amarum 57, herba Houttuyniae 170, Glycyrrhrizae radix 57 Gypsum Fibrosum 170.
The traditional Chinese medicine composition disclosed by the invention also preferably comprises the following raw material medicines in parts by weight:
255 portions of forsythia, 255 portions of honeysuckle, 255 portions of isatis root, 255 portions of rhubarb, 51 portions of patchouli, 85
Rhizoma dryopteris crassirhizomae 255 rhodiola rosea 85 menthol crystal 7.5 mix-fried ephedra 85
Parched semen Armeniacae amarum 85 herba Houttuyniae 255 Glycyrrhrizae radix 85 Gypsum Fibrosum 255.
The traditional Chinese medicine of the invention can be replaced by traditional Chinese medicines with the same or similar effects, and the traditional Chinese medicines can be processed according to national traditional Chinese medicine processing standard or traditional Chinese medicine dictionary.
The active ingredients of the traditional Chinese medicine composition are prepared by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
the dry paste powder obtained in the step (5), the volatile oil obtained in the step (2) and the menthol jointly form the active ingredients of the traditional Chinese medicine composition.
The medicament of the invention is in the form of capsules, tablets, powders, granules, oral liquid, soft capsules, pills, tinctures, syrups, suppositories, gels, sprays or injections.
In order to make the above dosage forms possible, pharmaceutically acceptable excipients, such as: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, bases, and the like. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crospolyvinylpyrrolidone, low-substituted hydroxypropylcellulose, croscarmellose sodium, etc.; the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like; the adhesive comprises starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, and the like; the flavoring agent comprises: sweeteners and various essences; the preservative comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its salts, benzalkonium bromide, chloroacetidine acetate, eucalyptus oil, etc.; the matrix comprises: PEG6000, PEG4000, insect wax, etc.
The capsule is prepared by the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, and cleaning and selecting;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after the oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), concentrating to obtain fluid extract with relative density of 1.15-1.20 at 60 deg.C, and drying to obtain dry extract powder;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the dry paste powder obtained in the step (5) for granulation;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the particles obtained in the step (6), sealing, uniformly mixing, and encapsulating to obtain the capsule.
The preparation method of the granules comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 5-8 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 6-10 times of 50-90% ethanol for 2 times, each for 1-3 hr, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 7-11 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot kernel, decocting for 2 times, 0.5-2.5 hours each time, combining the extracting solutions, filtering, combining the obtained filtrate with the filtrate obtained after oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding ethanol, adjusting the alcohol concentration to be 70%, refrigerating, standing, filtering, and recovering the ethanol until no alcohol smell exists, thus obtaining the clear paste for later use;
(5) mixing the fluid extract obtained in step (4) with the ethanol extract obtained in step (3), and concentrating to obtain soft extract with relative density of 1.25-1.35 at 60 deg.C;
(6) adding appropriate pharmaceutically acceptable adjuvants into the soft extract obtained in step (5), and granulating;
(7) and (3) dissolving the menthol and the volatile oil obtained in the step (2) in ethanol, spraying the granules obtained in the step (6), sealing, uniformly mixing and bagging to obtain the menthol-containing capsule.
The preparation method of the preferred granules comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the weight proportion of the raw materials, cleaning, and cutting off the traditional Chinese medicinal materials as appropriate;
(2) crushing herba Agastaches, extracting volatile oil with 6 times of water for 4 hr, and collecting volatile oil; filtering the extractive solution, discarding residue, and collecting filtrate;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 8 times of 70% ethanol for 2 times, 2 hr for the first time, and 1.5 hr for the second time, mixing extractive solutions, filtering, and recovering ethanol to obtain filtrate;
(4) adding 9 times of water into honeysuckle, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding fried bitter apricot seed, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, combining extracting solutions, filtering, combining obtained filtrate with water filtrate obtained after oil extraction of pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 measured at the temperature of 60 ℃, adding 95% ethanol, adjusting the alcohol concentration to be 70%, refrigerating, standing, filtering, and recovering ethanol until no alcohol smell exists, so as to obtain clear paste filtrate;
(5) mixing the fluid extract obtained in step (4) and the ethanol extract obtained in step (3), and concentrating to obtain soft extract with relative density of 1.25-1.35 at 60 deg.C;
(6) adding a proper amount of pharmaceutically acceptable auxiliary materials into the thick paste obtained in the step (5), granulating, and finishing granules for later use;
(7) and (3) screening out fine powder from the granules obtained in the step (6), adding ethanol into the menthol and the volatile oil obtained in the step (2) for dissolving, spraying the fine powder, uniformly mixing with the granules obtained in the step (6), sealing for half an hour, and bagging to obtain the menthol crystal.
The preparation method of other dosage forms of the medicine comprises the following steps: the raw materials are weighed according to the proportion and prepared by a conventional preparation method, for example, a preparation process recorded in Vanbitsin traditional Chinese medicine pharmacy (1 st 12 months in 1997 of Shanghai scientific Press) to prepare a conventional dosage form acceptable in pharmacy.
The invention also provides application of the traditional Chinese medicine composition in preparing a medicine for improving breathlessness and/or dyspnea.
The invention also provides application of the traditional Chinese medicine composition in preparing medicines for improving fever, cough and hypodynamia.
The invention also provides application of the traditional Chinese medicine composition in preparing a medicine for improving chest distress.
The invention also provides application of the traditional Chinese medicine composition in preparing a medicine for improving pulmonary damp-like pitch.
The invention also provides application of the traditional Chinese medicine composition in preparation of drugs for inhibiting increase of cytokines and chemokines.
The invention also provides the cytokine and the chemotactic factor which are TNF-alpha, IL-6, CCL-2/MCP-1, CXCL-10/IP-10.
The disease belongs to the category of pestilence in traditional Chinese medicine, pathogenic factors of epidemic toxin are main pathogenic factors, invasion of interior and heat dissipation are important pathogenesis outcome, and the disease is consistent with the clinical characteristics that pneumonia infected by novel coronavirus takes fever as main manifestation. The traditional Chinese medicine composition mainly comprises fructus forsythiae, honeysuckle, isatis root, rhubarb, patchouli, male fern rhizome, rhodiola rosea and the like. Is an innovative Chinese medicinal composition which is developed by applying the Chinese medicine collateral disease theory to reveal the infectious disease transmission rule of the respiratory system caused by viruses and taking 'antipyretic detoxification, lung diffusing and heat discharging' as a treatment method. The prescription takes the Yinqiao powder from Shangzhongjing Han Lun of Han Dynasty and Wujutong of Qingdai Jutong of Wen Bing tiao as a basic prescription, draws the experiences of Wu of Ming Dynasty and Wu Jutong of Wen Yi Lun of Wen Yi, combines rhodiola rosea with the experiences of clearing lung-heat and removing blood stasis and regulating immunity, embodies the medicine using experience of preventing and treating the epidemic diseases of the Chinese medicine for two thousand years, and particularly adjusts the prescription and the preparation method aiming at the pathogenesis and the expression symptoms of the pneumonia infected by the novel coronavirus, thereby being clinically applicable to infectious diseases of respiratory system accompanied with fever, chilliness, cough, muscle soreness and the like.
According to reports, the main clinical symptoms of pneumonia infected by the novel coronavirus can be fever, muscle pain, hypodynamia, expectoration, short breath, headache, pharyngalgia, watery nasal discharge, diarrhea, nausea and vomiting and the like.
Example 4:
the formula of the raw material medicine is as follows:
weeping forsythia 170 g honeysuckle flower 170 g mix-fried Chinese ephedra 57 g stir-fried bitter apricot kernel 57 g
170 g of gypsum, 170 g of isatis root, 170 g of male fern rhizome, 170 g of cordate houttuynia
Patchouli 57 g rhubarb 34 g rhodiola 57 g menthol 5.0 g
Licorice root, radix Glycyrrhizae 57 g
The preparation method comprises the following steps:
the extraction process comprises the following steps:
(1) weighing the traditional Chinese medicinal materials according to the prescription, cleaning, and cutting according to the requirement;
(2) extracting herba Agastaches with 6 times of water for 4 hr, collecting volatile oil with oil yield of 0.33%, filtering the extractive solution, removing residue, and collecting water extractive solution;
(3) extracting fructus forsythiae, herba Ephedrae preparata, herba Houttuyniae, and radix et rhizoma Rhei with 8 times of 70% ethanol for 2 times, 2 hr for the first time, and 1.5 hr for the second time, filtering the extractive solutions, mixing the filtrates, and recovering ethanol until no ethanol smell exists;
(4) adding 9 times of water into honeysuckle, fried bitter almond, gypsum, isatis root, male fern rhizome, liquorice and rhodiola rosea, decocting until boiling, adding the fried bitter almond, decocting for 2 times, wherein the first time is 1.5 hours, and the second time is 1 hour, filtering an extracting solution, combining filtrates, simultaneously adding an aqueous solution obtained after oil extraction of the pogostemon cablin in the step (2), concentrating into clear paste with the relative density of 1.10-1.15 determined at 60 ℃, adding 95% ethanol, stirring while adding until the alcohol concentration is 70%, refrigerating and standing for 24 hours, filtering, recovering ethanol from the filtrate until no alcohol taste exists, combining with an alcohol extracting solution, concentrating into thick paste with the relative density of 1.25-1.35 determined at 60 ℃ for later use;
(II) preparation process:
(5) the formula of the preparation is as follows: 335.5g of thick paste obtained in step (4) and 5g of menthol
0.2ml of patchouli oil obtained in the step (2) and 342.5g of powdered sugar 514.0g of dextrin
(6) And (3) granulating: mixing sugar powder and dextrin, making soft mass with soft extract as binder, granulating with 14 mesh screen, oven drying at 60-65 deg.C, and grading with 10 mesh screen;
(7) subpackaging: sieving to obtain fine powder, adding appropriate amount of ethanol into Mentholum and herba Agastaches volatile oil, dissolving, spraying into the fine powder, mixing with the granule, sealing for half an hour, and packaging to obtain 1000g granule.
Examples of the experiments
To confirm the efficacy of the pharmaceutical composition of the present invention for preventing and treating pneumonia caused by a novel coronavirus infection, the following clinical trial studies were conducted using the particles (hereinafter, LHQW or the pharmaceutical composition of the present invention) prepared in the manner of example 4:
experimental example 1:
in vitro cell study
Cell model research is carried out by professor team of Yang Zifeng in national emphasis laboratory of respiratory diseases at Guangzhou medical university aiming at LHQW anti-new coronavirus and anti-inflammatory activity, and the LHQW is found to have good inhibition effect on cytopathic effect caused by infection of common coronavirus (HCoV-229E) and novel coronavirus (2019-nCoV), IC50 is 338.74 and 414.02 mug/mL respectively, and SI is 3.47 and 2.89 respectively (Table 1).
The transmission electron microscope analysis shows that the normal cell has complete cell morphology, and no virus particles exist in extracellular cells; cells infected with viruses can see a large number of virus particles in cell membranes, cytoplasm and vesicles; intracellular virions were less in LHQW-treated cells than in virus-infected cells, with some virions being slightly distorted (fig. 1).
Adding LHQW with different concentrations after 2019-CoV infects Huh-7 cells, extracting cell mRNA after 48 hours, carrying out reverse transcription to form cDNA, and detecting the expression of inflammatory factors by an RT-qPCR method. The results show that the two drugs have obvious inhibition effects on mRNA expression of inflammatory factors such as TNF-alpha, IL-6, CCL2/MCP-1 and CXCL-10/IP-10 and the like, and have a dose-dependent relationship (figure 2).
Experimental example 2:
a data and method
1. Clinical data
And collecting the pneumonia diagnosis standard meeting the infection of the novel coronavirus, and judging the patient to be positive by performing nucleic acid detection on samples such as sputum, throat swabs, lower respiratory tract secretions and the like. Inclusion criteria were: ordinary hospitalized patients aged over 18 years, with pneumonia diagnosis standard of novel coronavirus infection and body temperature >37.2 ℃. Exclusion criteria: (1) patients with severe, and novel coronavirus infection pneumonia; (2) acute respiratory diseases caused by non-2019-nCoV; (3) any other chronic respiratory disease, respiratory bacterial infection such as suppurative tonsillitis, acute tracheo-bronchitis, sinusitis, otitis media and other respiratory diseases affecting the evaluation of clinical trials; (4) asthma needing daily treatment, and basic lung diseases such as severe pulmonary interstitial disease and bronchiectasis proved by chest CT; (5) with the basic diseases of serious primary immunodeficiency, acquired immunodeficiency syndrome, congenital respiratory deformity, congenital heart disease, lung dysplasia, etc.
2. The grouping method comprises the following steps: 21 common hospitalized patients with the age of more than 18 years old and the body temperature of more than 37.2 ℃ and meeting the pneumonia diagnosis standard of the novel coronavirus infection are taken as a treatment group, and the treatment scheme is conventional treatment (refer to a pneumonia prevention and control scheme of the novel coronavirus infection published by the national Weijian Commission) and the medicinal composition is added for 1 bag per time and 3 times per day; age and body temperature were used as covariates, and a logistic regression model was used to calculate the propensity score values, which were matched at a 1:1 ratio. The matched 21 patients served as a control group and the treatment regimen was conventional.
3. The evaluation indexes compare the disappearance rates of the main symptoms (fever, hypodynamia and cough), the disappearance time of the fever and other single symptoms of the treatment group and the control group.
4. Statistical methods statistical analysis used SAS 9.4 software. All statistical tests used a two-sided test, and a P value of 0.05 or less would be considered statistically significant for the differences tested. Descriptive analysis: the counting data is described by the number of cases and the composition ratio, and the metering data is described by the mean and the standard deviation. The group comparison of quantitative data adopts t test or classification data adopts chi-square test or accurate probability method.
Second, result in
1. Baseline data
Pneumonia with new coronavirus infection meeting requirements confirmed diagnosis of common type of patients total 42, treatment group male 16 (76.2%), female 5 (23.8%), average age (57.1 ± 14.0) year of week; control group male 12 cases (57.1%), female 9 cases (42.9%), average age (55.4 ± 12.3) year of week. The age and sex of the two groups of patients, as well as the body temperature, blood pressure, heart rate, respiration, past medical history, time from onset to diagnosis and other baseline data, compared among the groups, the difference has no statistical significance (P is more than 0.05), and the comparison is comparable and is shown in Table 2.
2. Disappearance of primary symptoms two results were compared: (1) baseline data: 21 cases in the treatment group, among which 21 cases of fever (100%), 15 cases of cough (71.4%), 12 cases of asthenia (57.1%); the control group comprises 21 cases with fever (100%), cough (18 cases with 85.7%), hypodynamia (13 cases with 61.9%), and the comparison difference between the groups has no statistical significance (P is more than 0.05). (2) The treatment results are as follows: compared with the control group, 18 cases (85.7%) of fever symptom disappeared in the treatment group and 7 cases (46.7%) of cough symptom disappeared, and the comparison difference between the groups has statistical significance (P < 0.05), which is detailed in Table 3.
3. Duration of heat generation two sets of results were compared: 21 cases of treatment, duration of fever (4.6 ± 3.2) d; the control group 21, duration of fever (6.1 ± 3.1) d, compared between groups had no statistical significance (P = 0.218).
4. Other symptom disappearance rates two sets of results were compared: compared with the control group, the expectoration rate and the symptom disappearance rate of the short breath of the treatment group are respectively 64.3 percent and 77.8 percent, and compared among the groups, the difference has statistical significance (P < 0.05), and the details are shown in the table 4.
In the research process, 42 patients with fever symptoms and pneumonia infected by novel coronavirus are included, and the patients are accompanied with symptoms of cough, hypodynamia, expectoration, myalgia, short breath, pharyngalgia, nausea, vomit, anorexia, diarrhea and the like in different degrees. In the treatment process, the surprising discovery shows that patients taking the granules of the traditional Chinese medicine composition of the invention in the treatment group can obviously relieve the clinical symptoms of fever, cough, expectoration, short breath and the like during the administration period, and the disappearance time of the fever is averagely shortened by 1.5 days compared with that of a control group, and the medicine also has the clinical advantage of improving the fever symptoms. Meanwhile, the medicine also shows a good trend in the aspect of improving symptoms of hypodynamia, myalgia, nasal obstruction and headache.
The patients selected in the study were patients with mild-moderate pneumonia infected with the novel coronavirus, and the study was designed to allow for better communication and feedback medication efficacy of mild-moderate patients. It does not mean that the patient is not allowed to take the medicine.
Experimental example 3:
1 clinical data
1.1 diagnostic criteria
Referring to "diagnosis and treatment of pneumonia infected by novel coronavirus" (fourth edition of trial "), there is any one of epidemiological histories of tourism, residence, exposure, and aggregation, and clinical manifestations of fever and/or respiratory symptoms, normal/decreased early leukocyte count/decreased lymphocyte count, and any 2 of NCP imaging characteristics, so that the suspected cases can be determined.
1.2 inclusion criteria
Hospitalized patients with age over 18 years and with NCP imaging characteristics, who meet the above-mentioned criteria for the diagnosis of suspected cases.
1.3 exclusion criteria
Patients with confirmed diagnosis of heavy, critical NCP; patients with bronchial asthma; chest X-ray Computed Tomography (CT) devices have confirmed the presence of patients with severe interstitial lung disease, bronchiectasis, and other basic lung diseases; with the basic diseases of serious immunodeficiency, congenital respiratory deformity, congenital heart disease, lung dysplasia, etc.
1.4 general data
101 suspected patients are collected, wherein 38 patients applying conventional treatment are taken as a control group; 63 patients with conventional therapy in combination with the drug of the present invention were treated. The age and sex of two groups of patients, and baseline data such as body temperature, blood pressure, heart rate, respiration, past medical history, fever, hypodynamia, cough, conventional treatment, main laboratory examination indexes and the like have no statistical significance (P is more than 0.05) in comparison and have comparability. See table 5.
2 method
2.1 methods of treatment
The two groups of patients are monitored for illness state, nutrition support treatment, symptomatic treatment, antiviral treatment, antibacterial treatment and the like are given, the main treatment medicine is moxifloxacin hydrochloride sodium chloride injection which is 0.4g and is produced by Chengdu Zhengkang pharmaceutical industry Limited company for 1 time per day, and the batch number is 3419111102; ganciclovir injection, 0.5g, 1 time/day, manufactured by Hubei Keyi pharmaceutical industry GmbH, lot number 191003; 2.5g of intravenous injection human immunoglobulin, 1 time/day, manufactured by Tai Pont, Guizhou, biological products, Inc., batch No. 201906026; ambroxol hydrochloride injection 30mg for 2 times per day, produced by Tianjin pharmaceutical industry group Co., Ltd, batch number 1906116; doxofylline for injection, 0.2g, 1 time/day, manufactured by Ruiyang pharmaceutical Co., Ltd, lot number 19082116; methylprednisolone sodium succinate for injection, 40mg, 1 time/day, manufactured by Liaoning Haishiki pharmaceutical Co., Ltd., batch No. 20191027.
Control group: the treatment is given alone.
Treatment groups: the medicine of the invention is combined on the basis of the treatment, 6 g/bag, 1 bag each time, 3 times a day. Clinical data were collected from patients 10 days after treatment
2.2 Observation indicators and methods
2.2.1 comparison of disappearance of the chief symptoms in two groups of patients: the disappearance rate of (fever, hypodynamia and cough), the disappearance time of fever and the disappearance rate of other single symptoms and signs (muscle pain, expectoration, nasal obstruction, watery nasal discharge, pharyngalgia, shortness of breath, chest distress, dyspnea, headache, nausea, vomiting, anorexia, diarrhea and wet rales) and the condition of exacerbation in the treatment process.
2.2.2 evaluation of safety: before and after treatment, blood routine, urine routine, stool routine, liver function and kidney function are detected.
2.3 statistical methods
The statistical analysis employed SAS 9.4 software. All statistical tests adopt bilateral test, counting data of descriptive analysis adopts case number and composition ratio description, and metering data adopts average number +/-standard deviation description. The comparison between groups of the measurement data adopts t test, and the counting data adopts chi-square test or accurate probability method. The duration of fever was analyzed by survival. P.ltoreq.0.05 indicates that the difference is statistically significant.
3 results
3.1 comparison of disappearance rates of chief complaints in two groups of patients
Table 6 shows that compared with the control group, the symptoms of fever, cough and hypodynamia disappear in the treatment group obviously better than the control group (P is less than 0.05).
The treatment group comprises 52 patients with fever, and the median fever duration is 6 days; the control group had 23 total febrile patients: median duration of fever 7 days, two comparative groups had no statistical significance (P = 0.171).
3.3 comparison of disappearance rates of other symptoms and signs in two groups of patients
Table 7 shows that the tachypnea symptoms and the rate of rales are 68.2% and 56.0% respectively, which are significantly better than 20.0% and 20.0% of the control group (2 =9.817, 4.972), and the comparative differences of the two groups have statistical significance (P < 0.05); no statistical difference was observed in the comparison of the disappearance rates of other symptoms between the two groups.
3.4 exacerbation of disease in the course of treatment of two groups of patients
In 63 cases of the treatment group, the disease condition is aggravated in 4 cases (6.4%) during the treatment process; in the control group 38 cases, the disease was increased by 6 cases (15.8%) during the treatment, and there was no statistical difference (P > 0.05) between the two groups.
3.5 safety assay
In the treatment process, the abnormal conditions related to the medicine do not appear in the routine treatment group blood and laboratory examinations of liver and kidney functions and the like, the adverse reaction of the medicine does not appear, and the safety of clinical application is good
Experimental example 4:
1 object and method
1.1 the study subjects collected patients, and the patients were judged to be positive NCP patients according to the general diagnosis standard of diagnosis and treatment protocol for pneumonia infected by novel coronavirus (trial fifth edition), and 54 patients who were treated with the drug of the present invention in combination with conventional therapy (nutrition support therapy, symptomatic therapy, antiviral therapy, and antibacterial therapy according to disease monitoring) were tested by nucleic acid detection using samples such as sputum, throat swab, and lower respiratory secretion.
1.2 research method analysis 54 NCP common type patients clinical data, including the main symptoms (fever, hypodynamia, cough) in 3 days, 5 days, 7 days of disappearance, fever disappearance days and other symptoms sign disappearance rate to retrospectively summarize analysis, and meanwhile with the main symptoms "no" to count 0 points, "yes" to count 1 points, symptoms score decrease rate > 30% to judge treatment effective, less than 30% to judge treatment ineffective, analysis after 7 days of treatment all patients treatment effective rate.
2 results
2.1 general data 54 hospitalized patients aged 25-95 years, mean (60.1 + -16.98) years; of which 29 cases (53.7%) in men and 25 cases (46.3%) in women; average body temperature (37.93 + -0.93) deg.C, and median temperature 38.05 deg.C; maximum body temperature (38.54 + -0.60) deg.C before diagnosis; heart rate is average (87.9 +/-11.80) times/minute, median times are 85.5 times/minute, and highest 112 times/minute; average (21.1 +/-3.78) breaths per minute, the median frequency is 20.0 times per minute, and the highest is 30.0 times per minute; 21 cases of previous hypertension (38.9%), 7 cases of coronary heart disease (13.0%), 10 cases of diabetes (18.5%), 10 cases of cerebral infarction (18.5%); laboratory examination: 31 of 54 patients with leukocytes in the normal range (64.6%), 9 below normal (18.8%), and 8 above normal (16.7%); neutrophils 25 in the normal range (52.1%), 23 higher than normal (47.9%); 14 cases (29.2%) of lymphocytes within the normal range were lower than 34 cases (70.8%) of normal; the hypersensitive C reactive protein is higher than the normal value (100%); all patients applied with the medicament of the invention have the average days (8.0 +/-4.10) days, the median days are 7.0 days, the shortest day is 1.0 day, and the longest day is 16.0 days.
2.2 disappearance rate of main symptoms (fever, hypodynamia and cough): 40 of 54 patients suffered from fever (74.1%), fatigue (37) and cough (55.6%); ② after 3 days of treatment, 19 cases of fever symptom disappear (disappearance rate 47.5%), 13 cases of hypodynamia symptom disappear (disappearance rate 35.1%), 6 cases of cough symptom disappear (disappearance rate 20.0%); ③ after 5 days of treatment, 25 cases of fever symptoms disappeared (disappearance rate 62.5%), 22 cases of hypodynamia symptoms disappeared (disappearance rate 59.5%), and 15 cases of cough symptoms disappeared (disappearance rate 50.0%); after 7 days of treatment, 32 cases of fever symptoms disappear (disappearance rate 80.0%), 28 cases of hypodynamia symptoms disappear (disappearance rate 75.7%), and 23 cases of cough symptoms disappear (disappearance rate 76.7%). See table 9.
2.3 days for disappearance of main symptoms (fever, hypodynamia and cough), namely, the average days for disappearance of fever symptoms (3.6 +/-2.14) days, 3.0 days for disappearance of median, 1.0 day for disappearance of shortest and 8.0 days for disappearance of longest; ② the average disappearance days (4.1 +/-2.58) of the hypodynamia symptoms, 4.0 days of median disappearance, shortest 1.0 disappearance and longest 12.0 disappearance; ③ average disappearance days (5.3 +/-2.63) of cough symptoms, 5.0 median disappearance days, shortest disappearance of 1.0 day and longest disappearance of 12.0 days.
2.4 disappearance of other signs of symptoms (before treatment): 13 patients with chest distress (24.1%), 8 patients with dyspnea (14.8%), 10 patients with anorexia (18.5%), and 19 patients with wet rales (35.2%); ② after 7 days of treatment: 2 cases with chest distress symptom and 11 cases of disappearance (disappearance rate 84.6%); dyspnea symptoms 0, 8 disappeared (disappearance 100%); symptoms of anorexia 6, disappearance 4 (40.0%); 2 examples of damp-warm signs and 17 examples of disappearance (disappearance rate 89.5%). See table 10.
2.5 therapeutic effectiveness 3 days later, 23 of 54 NCP patients were effective (effective rate 46.9%); after 5 days of treatment, 34 effective cases (the effective rate is 69.4%); after 7 days of treatment, 40 effective cases (the effective rate is 81.6%) are shown in table 11.
2.6 safety analysis in the course of applying the medicine of the present invention in combination with conventional treatment, no abnormal condition related to the medicine of the present invention appears in laboratory examinations of blood routine, liver and kidney functions, etc., no adverse reaction of the medicine of the present invention appears, and the safety of clinical application is good.
Experimental example 5:
1 object and method
1.1 clinical data of patients who are diagnosed with the NCP are collected from the study subjects, and the clinical data of the patients are judged to be positive by nucleic acid detection through samples such as sputum, throat swabs, lower respiratory tract secretions and the like.
1.2 the patients are included between the ages of 18 and 70, and the patients meet the diagnosis standard of NCP common type, and are hospitalized for more than 6 days.
1.3, removing the standard of heavy type and dangerous type NCP patients; ② any other chronic respiratory diseases, respiratory system bacterial infection such as suppurative tonsillitis, acute tracheo-bronchitis, sinusitis, otitis media and other respiratory diseases which affect the evaluation; ③ basic diseases such as serious pulmonary interstitial disease, bronchiectasis, primary immunodeficiency disease, congenital respiratory deformity, congenital heart disease, lung dysplasia, etc.; patients with serious liver diseases (glutamic-oxaloacetic transaminase AST and glutamic-pyruvic transaminase ALT exceed the upper limit value of the normal by 5 times), or serious renal insufficiency or receiving continuous renal replacement therapy, hemodialysis and peritoneal dialysis; the patient has many metastases and can not be resected, such as malignant tumor, hematopathy, dyscrasia, active hemorrhage, severe malnutrition, HIV, etc., or serious neurological and psychiatric diseases.
1.4 grouping method collects 51 patients meeting the nano-grade standard and having more than or equal to 5 days of treatment course of the medicine, then age, body temperature and course of disease are used as covariates, Logistic regression model is used for calculating tendency score value, and 51 patients are matched as a control group in the conventional treatment group according to the proportion of 1: 1.
1.5 methods of treatment control group: simple nutrition support treatment, symptomatic treatment, antiviral treatment and antibacterial drug treatment. Treatment groups: the drug of the invention is combined on the basis of a control group, 6 g/bag, 1 bag each time, 3 times a day. Clinical data were collected from patients treated for 7 days.
1.6 the observation indexes analyze the clinical data of two groups of patients, including the disappearance rate and disappearance days of main symptoms (fever, hypodynamia and cough), the disappearance rate of other symptoms and signs, the effective rate of the main symptoms, the CT conversion rate, the clinical conversion rate and the like.
1.7 evaluation criteria (rate of disappearance of symptom): the number of cases/total number of cases in which symptoms disappeared after treatment was the rate of symptom disappearance; effective rate of main symptoms: the main symptoms (fever, hypodynamia and cough) are not counted by 0 point, the main symptoms (fever, hypodynamia and cough) are counted by 1 point (before treatment, after treatment) and before treatment, the symptom integral reduction rate is judged to be effective when the symptom integral reduction rate is more than 30 percent, the treatment is judged to be ineffective when the symptom integral reduction rate is less than or equal to 30 percent, and the number of cases/the total number of cases judged to be effective after the treatment is the effective treatment rate. ③ CT improvement rate: post-treatment pulmonary imaging (CT) showed a CT improvement rate of cases/total cases over the previous improvement; fourthly, clinical weight transfer rate: referring to the standard of heavy diagnosis in diagnosis and treatment of pneumonia infected by novel coronavirus (trial fifth edition), the number of cases with normal type converted to heavy type/total number of cases was the clinical conversion rate.
1.8 statistical analysis of statistical method adopts SAS 9.4 software, all statistical tests adopt bilateral test, counting data of descriptive analysis adopts case number and composition ratio description, and metering data adopts mean ± standard deviation description. The comparison between groups of the measurement data adopts t test, and the counting data adopts chi-square test or accurate probability method. The duration of fever was analyzed by survival. P.ltoreq.0.05 indicates that the difference is statistically significant.
2 results
2.1 general data
A total of 102 patients with satisfactory confirmed normal type of diagnosis were collected, and 51 treatment groups were selected, including 26 cases in men (51.0%), 25 cases in women (49.0%), mean (55.5 ± 12.3) years of age, and mean body temperature (38.44 ± 0.63) ° c; systolic pressure (122.6 + -11.9) mmHg, diastolic pressure (75.0 + -9.2) mmHg; heart rate (89.0 + -11.6) times/min and respiration (19.8 + -2.4) times/min. 15 cases of previous hypertension (29.4%), 5 cases of coronary heart disease (9.8%), 4 cases of diabetes (7.8%), 3 cases of cerebral infarction (5.9%); laboratory examination: 13 cases of blood leukocyte reduction (25.5%), 20 cases of lymphocyte reduction (39.2%), 40 cases of hypersensitive C-reactive protein increase (78.4%), 20 cases of blood sedimentation increase (39.2%), 16 cases of procalcitonin increase (31.4%);
a control group of 51 cases, in which 27 cases (52.9%) of men, 24 cases (47.1%) of women, mean (55.8 ± 11.6) years of age, mean body temperature (38.33 ± 0.64) ° c, systolic blood pressure (127.0 ± 16.3) mmHg, diastolic blood pressure (74.8 ± 10.3) mmHg; heart rate (88.7 +/-13.4) times/minute, respiration (20.0 +/-2.8) times/minute, 17 cases of previous hypertension (33.3%), 2 cases of coronary heart disease (3.9%), 4 cases of diabetes (7.8%) and 6 cases of cerebral infarction (11.8%); laboratory examination: 16 cases of blood leukocyte reduction (31.4%), 24 cases of lymphocyte reduction (47.1%), 42 cases of hypersensitive C-reactive protein increase (82.4%), 24 cases of blood sedimentation increase (47.1%), and 17 cases of procalcitonin increase (33.3%).
The age and sex of two groups of patients, and baseline data such as body temperature, blood pressure, heart rate, respiration, past medical history, laboratory test results and the like have no statistical significance (P is more than 0.05) in comparison between the two groups, and have comparability. See table 12.
Note: male, past medical history and laboratory examination results are counting data, and chi fang inspection and comparison are adopted; the rest is measured data, and t test comparison is adopted. 1 mmHg. apprxeq.0.133 kPa.
2.2 comparison of disappearance rates of chief complaints in two groups of patients
Firstly, baseline data: treatment group 51 cases, of which 43 cases (84.3%) had fever, 31 cases (60.8%) had asthenia, and 37 cases (72.6%) had cough; a control group of 51 cases, in which fever was 41 cases (80.4%), fatigue was 35 cases (68.6%), cough was 39 cases (76.5%); comparison of differences between groups was not statistically significant. ② after 7 days of treatment, 36 cases (83.7%) of fever symptom disappeared in the treatment group compared with the control group, which was significantly better than 25 cases (61.0%) in the control group (x 2=5.461, P = 0.019); 19 cases (61.3%) of the debilitating symptoms disappeared, significantly better than 12 cases (34.3%) of the control group (x 2=4.813, P = 0.028); cough symptoms disappeared 23 cases (62.2%), significantly better than 14 cases (35.9%) of the control group (x 2=5.243, P = 0.022); the comparative differences between the groups were statistically significant (P < 0.05). See table 13.
2.3 comparison of disappearance time of chief complaints in two groups of patients
The treatment group has 36 patients with fever disappearance, the average fever duration is (2.9 +/-1.67) days, the asthenia disappears 19 cases, the average duration is (3.5 +/-1.50) days, the cough disappears 23 cases, and the average duration is (3.9 +/-1.98) days; the control group contains 25 patients with fever disappeared, the average duration is (3.9 + -1.29) days, the lack of strength disappeared 12 cases, the average duration is (4.8 + -1.53) days, the cough disappeared 14 cases, and the average duration is (5.2 + -1.76) days. The disappearance time of the main symptoms (fever, hypodynamia and cough) of the patients in the treatment group is shorter than that of the patients in the control group, and the comparative difference among the groups has statistical significance (P is less than 0.05). See table 14.
2.4 effective rate comparison of main symptoms of two groups of patients
After 7 days of treatment, the main symptoms of 51 patients in the treatment group are effectively treated by 44 patients, and the effective rate is 86.3%; in 51 patients in the control group, the treatment of the main symptoms is effective for 35 patients, the effective rate is 68.6%, and the comparison difference among the groups has statistical significance (P = 0.033).
2.5 comparison of disappearance rates of other symptom signs in two groups of patients
Firstly, baseline data: 51 cases of treatment group, among which 9 cases of myalgia (17.7%), 20 cases of expectoration (39.2%), 13 cases of breathlessness (25.5%), 11 cases of chest distress (21.6%), 3 cases of dyspnea (5.9%), 7 cases of nausea (13.7%), 23 cases of anorexia (45.1%), 22 cases of pulmonary wet rales (43.1%); 51 cases of the treatment group, among which 11 cases of myalgia (21.6%), 19 cases of expectoration (37.3%), 14 cases of breathlessness (27.5%), 19 cases of chest distress (37.3%), 7 cases of dyspnea (13.7%), 5 cases of nausea (9.8%), 26 cases of anorexia (51.0%), 23 cases of pulmonary moist rales (45.1%); comparison of differences between groups was not statistically significant (P > 0.05). (II) after 7 days of treatment, 11 cases (55.0%), 8 cases (61.5%), 6 cases (54.6%), 8 cases (34.8%), 10 cases (45.5%), 3 cases (15.8%), 2 cases (14.3%), 3 cases (15.8%), 2 cases (7.7%) and 3 cases (13.0%) of the treatment group disappear, and the comparison difference between the two groups has statistical significance (P < 0.05); there were no statistical differences in comparison between the treatment groups (P > 0.05) for 6 cases of myalgia, dyspnea, nausea disappearance (66.7%), 2 cases (66.7%), 4 cases (57.1%), 2 cases of disappearance (18.2%), 2 cases (28.6%), 2 cases (40.0%) in the control group. See table 15.
2.6 analysis of the severity of the metastases in the course of treatment of two groups of patients
Treatment group 51, 4 of the transponding type (7.8%) during the treatment; the comparison between the control group, 51, and the weight of the transgenic animal (21.6%) and the group was statistically significant (P < 0.05).
2.7 comparison of the improvement rates of two Lung imaging (CT)
After 7 days of treatment, lung CT showed 28 improvement (54.9%) in 51 cases of the treatment group; in the control group, there were 23 lung CT improvement (45.1%) in 51 cases, and the comparison between the groups showed no statistical significance (P > 0.05).
The traditional Chinese medicine and western medicine are combined to treat the novel coronavirus patient, the treatment is not only equivalent to combined medication, but also is carried out by respective conventional functional indications through a conventional treatment method, and the treatment is only a simple superposition. The traditional Chinese medicine composition has obvious treatment effect by combining with conventional treatment of western medicines under the condition that various antiviral medicines can not treat and prevent novel coronavirus due to the integral regulation capacity, and the test result shows that the symptoms of the common early and middle-stage patients are obviously improved, the patients are easy to recover after treatment, the severe transformation of the middle-stage patients is obviously reduced, and the non-severe transformation of the patients are effectively controlled. After critical patients take the pharmaceutical composition provided by the invention, the blood oxygen saturation is stabilized, the dyspnea is improved, the inflammation absorption of the lung is promoted, and sequelae such as interstitial pneumonia and pulmonary fibrosis are avoided. The medicine of the invention is shown to obviously improve clinical symptoms and signs related to diseases such as fever, cough, hypodynamia, short breath, lung damp and the like; meanwhile, the proportion of the treatment group to the severe case also shows an obvious downward trend, and the medicament has good clinical curative effect on suspected cases, helps to improve clinical symptoms, relieves the severity of diseases and the like. The characteristic advantages of 'integral regulation and multi-target treatment' of the compound traditional Chinese medicine are revealed, and the application of the medicine provided by the invention in suspected cases is prompted to have important clinical application value.
Experimental example 6
In vitro cell study
1. Materials and methods
1.1 cell lines and viruses: african green monkey kidney epithelial cells (Vero E6) and the human hepatoma cell line Huh-7 were purchased from ATCC. The culture conditions are as follows: dulbecco's modified Eagle's Medium (DMEM, Gibco, USA) containing 10% Fetal Bovine Serum (FBS). The culture temperature is as follows: 37 ℃ is carried out. The novel coronavirus (SARS-CoV-2) is a clinical isolate from the first Hospital affiliated with Guangzhou medical university.
1.2 test article: LHQW, source: shijiazhan, Ling pharmaceutical products, Inc. Preparing and storing a sample stock solution: the test sample was dissolved in dimethyl sulfoxide (DMSO) and stored at-20 ℃. Reddeevir (Remdesivir) was supplied by professor Zhang Jiancun, Guangzhou institute for biomedical and health, Chinese academy of sciences, and Reddeevir was dissolved in DMEM medium.
1.3 cytotoxicity assay: the cytotoxic effect of LHQW on Vero E6 cells and Huh-7 cells was evaluated by the MTT method. Monolayers of Vero E6 cells and Huh-7 cells grown in 96-well plates were discarded from stock solutions, washed with PBS, and incubated for 72 hours with different concentrations of test solutions. After 72 hours, cells were stained with 0.5 mg/mL MTT solution for 4 hours. The supernatant was discarded, and the formazan crystals formed were dissolved in 200 μ L dimethyl sulfoxide (DMSO). Absorbance at 570 nm was measured using a full wavelength microplate reader (Thermo Fisher, USA).
1.4 cytopathy inhibition assay: vero E6 cells were seeded in a 96-well plate and after growth to a monolayer, a coronavirus strain with a titer of 100 TCID50 was seeded and cultured at 37 ℃ for 2 hours. The culture solution was discarded, LHQW solution or positive control, ridciclovir, was added at different concentrations and incubated for 72 hours. After 72 hours, infected Vero E6 cells showed 100% cytopathic effect under the microscope. The percentage of cytopathic effect in LHQW treated cells was recorded. The LHQW half inhibitory concentration (IC 50) on virus-induced cytopathic effects was calculated.
1.5 plaque reduction experiment: vero E6 cells were cultured to a monolayer in 6-well plates, the supernatant discarded, washed with PBS, and 50PFU of SARS-CoV-2 virus was added to the plates and incubated for 2 hours. The supernatant was discarded, basal medium containing 1.2% agarose (Invitrogen, usa) per ml and various concentrations of LHQW or the positive control, ridcoxib, were added, and after incubation for 48 hours at 37 ℃, 4% formalin was fixed for 20 minutes and 1% crystal violet was stained for 3 minutes. And calculating the plaque inhibition rate. The median inhibitory concentration of drug against the virus was calculated (IC 50).
1.6 RT-qPCR method: huh-7 monolayers seeded in 12-well plates were washed with PBS and infected with coronavirus at MOI =1 for 2 hours at 37 ℃. The supernatant was discarded, and the sample was diluted to the desired concentration with a medium containing 2% FBS, added to a 12-well plate, and incubated continuously for 48 hours. The collected cells were used for RNA isolation and qPCR detection. The primer and probe sequences used for the analysis are shown in Table 16. Relative mRNA expression was calculated using the 2- Δ Ct method using GAPDH as an internal reference.
1.7 Electron microscopy analysis: cells were fixed, dehydrated and embedded. The embedded wax blocks were cut into ultrathin 70nm sections. Stained with uranyl acetate and lead citrate and read under a JEM-1400 PLUS transmission electron microscope.
2. Statistical analysis
Statistical analysis was performed using GraphPad Prism 7.0 software. Differences in proinflammatory cytokine mRNA expression levels of each group were compared using one-way analysis of variance (ANOVA). The level of statistical significance was set at p < 0.05.
3. Results
3.1 in vitro antiviral Activity of LHQW against SARS-CoV-2
Cell viability of Vero E6 cells after treatment with LHQW or redciclovir was determined by MTT assay. At a concentration of 600. mu.g/mL, LHQW was significantly cytotoxic to VeroE6 cells (see FIG. 4A for results). The positive control, ridciclovir, was not cytotoxic to cells at a concentration of 50 μ M (results are shown in figure 4B).
As shown in FIG. 5A, LHQW was tested for inhibition of SARS-CoV-2 virus by the cytopathic inhibition assay and the plaque reduction assay, and had an IC50 value of 411.2. mu.g/mL (see FIGS. 5A, 5C). The IC50 value for Reidesciclovir against SARS-CoV-2 virus is 0.651. mu.M.
Vero E6 cells were infected with the virus 48 hours later, and virions were found in the cytoplasm, intracellular vesicles, endoplasmic reticulum, and cell membrane, and a typical coronavirus morphology was observed under an electron microscope (see FIGS. 6B, 6F). After LHQW (600. mu.g) treatment, the virus expression was reduced (see FIGS. 6F, 6G). More significantly, LHQW-treated cells altered virus morphology: many oval-shaped virions are present on the cell surface, unlike spherical, cuspated virions in virus-infected cells (see FIG. 6H).
3.2 inhibition of SARS-CoV-2-induced cytokine and chemokine expression by LHQW
To determine the role of LHQW in inhibiting SAR2-CoV-2 induced cytokine and chemokine expression, mRNA expression levels of TNF- α, IL-6, CCL-2/MCP-1 and CXCL-10/IP-10 in cells were determined after LHQW treatment. The results show that the expression of cytokines and chemokines is increased after 48 hours of virus infection, LHQW has obvious inhibition effect on the cytokines and chemokines, and the inhibition effect has concentration dependence. The results are shown in FIG. 6.
Conclusion
LHQW is a broad-spectrum antiviral preparation, has good curative effect on various respiratory tract virus infections, and is widely applied clinically. Based on previous research results, LHQW has good curative effect on various influenza viruses by interfering virus and host reaction. Although LHQW can significantly alleviate the clinical symptoms of the new coronavirus pneumonia, mechanisms for anti-coronavirus (especially highly pathogenic coronavirus) continue to be explored. In this study, we demonstrated that LHQW exerts anti-coronavirus activity by inhibiting virus replication and reducing cytokine release from host cells. In clinical applications, LHQW can be used alone or in combination with existing therapies to treat novel coronavirus pneumonia.