CN101361941A - Medicine combination for preventing and treating cold and preparation method thereof - Google Patents
Medicine combination for preventing and treating cold and preparation method thereof Download PDFInfo
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- CN101361941A CN101361941A CNA2008101988944A CN200810198894A CN101361941A CN 101361941 A CN101361941 A CN 101361941A CN A2008101988944 A CNA2008101988944 A CN A2008101988944A CN 200810198894 A CN200810198894 A CN 200810198894A CN 101361941 A CN101361941 A CN 101361941A
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Abstract
The invention provides a medicinal composition for controlling cold, influenza and other viral infectious diseases and a preparation method thereof. The medicinal composition for controlling cold consists of the ingredients with the following weight proportion: 0.5-20 portions of oil of zedoary turmeric, 5-50 portions of oil of patchouli, and 5-50 portions of supercritical carbon dioxide extract of Matricaria indica. The medicinal composition has the effects of eliminating wetness-evil with drugs of fragrant flavour and facilitating perspiration and detoxification, and pharmaceutical research reveals that the medicinal composition has the functions of antibiosis, antivirus, anti-inflammation, relieving heat and immunoregulation, and can be used for preventing and treating cold, influenza and other viral infectious diseases.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition, relate in particular to a kind ofly be applied to anti-cure cold, grippal medicine.
Background technology
Flu is modal a kind of respiratory infectious disease, wherein 70%～80%, cause that by viral infection small part is by due to the bacterial infection.Flu is endangering human beings'health, and it can reduce the immunocompetence of human body, causes other diseases, as pharyngolaryngitis, sinusitis, otitis media, bronchitis and pneumonia, even empyema, liver abscess, pericarditis and osteomyelitis etc.Owing to still do not have definite antipathogen medicine at present,, prevent secondary infection so Western medicine mainly is a relief of symptoms to the treatment of catching a cold.Kind surplus the Coritab of clinical practice existing 20, they have for the caused various symptoms of flu alleviates or the elimination effect, but finally also need lean on people's autoimmunity to cure, and the saliva of having a liking for, side effect such as sleepy are generally all arranged.
Influenza is a kind of even more serious than the common cold acute respiratory infectious disease that is caused by influenza virus, and its characteristics are that onset is anxious, infectiousness is strong, sickness rate is high, the course of disease is long.The effective means of preventing and treating influenza at present is the inoculation influenza vaccines, but because the influenza virus variability is strong, can make existing vaccine failure, thereby has strengthened the difficulty of control.
From the angle of the traditional Chinese medical science, flu, influenza are that affection of exogenous wind-cold causes on the basis in the body endogenous fire volt.Treatment by Chinese herbs flu both can be removed interior-heat, can evacuate the heresy of appearance wind and cold again, and treating both the exterior and interior at the same time, and the immune function that Chinese medicine can integrally-regulated human body reach and set upright the purpose of getting rid of evils, and can make flu, influenza obtain thoroughly effectively treating.The Chinese patent medicine of existing clinically tens of kinds of treatment flu comes out, as Radix Isatidis granule, antivirus oral liquid, Antiphlogistic Powder of Schizonepetae and Ledebouriellae etc., but because their medicine is formed comparatively single, indication relatively limits to, add that preparation process is relatively backward, onset is slower, and clinical efficacy is not ideal.Therefore, research and develop out evident in efficacy, toxic and side effects is low anti-ly cures cold, the influenza product, have great importance.
Summary of the invention
Problem at prior art exists the objective of the invention is to overcome above-mentioned the deficiencies in the prior art part, provides a kind of and is used for anti-ly curing cold, the drug regimen and the preparation method of influenza and other viral infection diseases.
The present invention adopts Oleum Curcumae, patchouli oil and Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing with antibiotic, antiviral and anti-inflammatory efficacy as main component, makes peroral dosage form.Oleum Curcumae is the volatile oil that extracts from zingiberaceous plant Rhizoma Curcumae, RADIX CURCUMAE, Guangxi zedoary, and main component is the hemiterpene class, as curcumenol, curzerenone, curdione etc.Modern pharmacological research shows that Oleum Curcumae has effects such as antibiotic, antiviral, antitumor, immunomodulating, antiinflammatory, and zoopery proof Oleum Curcumae has direct inhibitory action to respiratory syncytial virus, influenza virus A 1 type and A3 type.Now make Oleum Curcumae injection and Zedoary turmeric oil glucose injection and be used for the treatment of viral infection diseases such as upper respiratory tract infection, viral pneumonia clinically.But owing to Chinese medicine complex chemical composition, preparation technology fall behind, the untoward reaction of Oleum Curcumae injection happens occasionally, even causes death, has had a strong impact on Oleum Curcumae application clinically.Patchouli oil has the bacteriostatic effect as the main component of Herba Pogostemonis medical material, and pathogen such as staphylococcus aureus, alpha streptococcus are had certain inhibitory action.In addition, effects such as that bibliographical information Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing has is antibiotic, analgesic, enhance immunity now existingly are used for the treatment of the toothpaste of periodontal disease with it as raw material.
For reaching above-mentioned purpose, the anti-drug regimen that cures cold of the present invention, adopt following technical scheme:
A kind of anti-drug regimen that cures cold is made up of following materials of weight proportions medicine: 0.5～20 part of Oleum Curcumae, 5～50 parts of patchouli oils, 5～50 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things.
Further, the weight proportion of crude drug of the present invention is preferably: 2.7 parts of Oleum Curcumae, 13.5 parts of patchouli oils, 13.5 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things;
Further, described pharmaceutical composition and extract or refining thing can add conventional adjuvant or excipient, make clinical acceptable forms, are peroral dosage form; Peroral dosage form is selected from a kind of in the middle of pill, tablet, capsule, spray and the drop pill;
The application of above-mentioned pharmaceutical composition aspect respiratory system disease is used to catch a cold, the prevention and the treatment of influenza and other viral infections.
The source of crude drug of the present invention is: Oleum Curcumae is the fat-soluble part of the dry rhizome of zingiberaceous plant Rhizoma Curcumae Curcuma phaeocaulisValeton, RADIX CURCUMAE Curcuma wenyujin Y.H.Chen et C.Ling, Guangxi zedoary Curcumakwangsiensis S.G.Lee et C.F.Liang through steam distillation or the extraction of application supercritical carbon dioxide extraction method.Patchouli oil is the fat-soluble part of the aerial parts of labiate Herba Pogostemonis Pogostemoncablin (Blanco) Banth. through steam distillation or the extraction of application supercritical carbon dioxide extraction method.Flos Chrysanthemi Indici is the dry capitulum of feverfew Herba Dendranthematis indici Chrysanthemum indicum L..Quality standard: one one of Pharmacopoeia of the People's Republic of China version in 2005.
A kind of preparation method of the anti-drug regimen that cures cold comprises the steps:
1., at first prepare Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing, get the Flos Chrysanthemi Indici crude drug, be ground into coarse powder, put in the supercritical carbon dioxide extraction instrument and extract, the extraction parameter is 45 ± 3 ℃ of extraction temperature, extracting pressure 20 ± 10MPa, parsing pressure is 6～7MPa, 55 ± 5 ℃ of resolution temperatures, carbon dioxide flow is 25 ± 3L/h, the extraction time is 2 ± 1h, and the gained extract is put in the clean container, and is standby;
2., again with the Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing of above-mentioned preparation, respectively become weight proportion by drug regimen: 0.5～20 part of Oleum Curcumae, 5～50 parts of patchouli oils, 5～50 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things, above material mixing stirs, and makes drug regimen then.
The present invention is a kind of drug regimen that has eliminating turbid pathogen with aromatics, delivers detoxicating functions, and pharmacological research shows that it has antibiotic, antiviral, antiinflammatory, analgesic and reconcile the effect of immunity.And this drug regimen antiviral, antiinflammatory and refrigeration function all are eager to excel with Oleum Curcumae, patchouli oil or Wild chrysanthemum extract than independent.Can be used for catching a cold, the prevention and the treatment of influenza and other viral infections.
The specific embodiment
For further understanding feature of the present invention, technological means and the specific purposes that reached, function, will describe in further detail the present invention by zoopery below.
At first prepare Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing, get the Flos Chrysanthemi Indici crude drug, be ground into coarse powder, put in the supercritical carbon dioxide extraction instrument and extract, the extraction parameter is 45 ± 3 ℃ of extraction temperature, extracting pressure 20 ± 10MPa, parsing pressure is 6～7MPa, 55 ± 5 ℃ of resolution temperatures, carbon dioxide flow is 25 ± 3L/h, the extraction time is 2 ± 1h, and the gained extract is put in the clean container, and is standby; With the Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing for preparing, respectively become weight proportion by drug regimen: 0.5～20 part of Oleum Curcumae, 5～50 parts of patchouli oils, 5～50 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things, above material mixing stirs, and makes drug regimen then.
Example 1 (hard capsule):
Get 2.7g Oleum Curcumae, 13.5g patchouli oil, 13.5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing,, get clathrate with 240g betacyclodextrin enclose; Add starch to 370g, mixing is made granule, and is dry below 60 ℃, sieves, and the capsule of packing into No. 1 is made 1000 of hard capsules (0.37g/ grain).Taking dose is: every day 2 times, each 2.
Example 2 (tablet):
Get 2.7g Oleum Curcumae, 13.5g patchouli oil, 13.5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing,, get clathrate, add starch to 250g with 240g betacyclodextrin enclose, mixing is made granule, and is dry below 60 ℃, be pressed into 1000 (0.25g/ sheets), the bag film-coat, promptly.Taking dose is: every day 2 times, each 2.
Example 3 (soft capsule):
Get 2.7g Oleum Curcumae, 13.5g patchouli oil, 13.5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing, add Oleum Arachidis hypogaeae semen to 250g, mixing is made 1000 of soft capsules (0.25g/ grain).Taking dose is: every day 2 times, each 2.
Example 4 (drop pill):
Get the 170g Macrogol 4000,70 ℃ of heating in water bath make fusion, add 2.7g Oleum Curcumae, 13.5g patchouli oil, 13.5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing, and mixing splashes in the refrigerative simethicone, makes drop pill 1000 balls (0.21g/ ball).Taking dose is: every day 2 times, each 2 balls.
Example 5 (granule):
Get 2.7g Oleum Curcumae, 13.5g patchouli oil, 13.5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing, with 240g betacyclodextrin enclose, get clathrate, add starch to 250g, mixing is made granule, and is dry below 60 ℃, is distributed into 500 bags (1g/ bags).Taking dose is: every day 2 times, each 1 bag.
Example 6 (soft capsule):
Get 20g Oleum Curcumae, 50g patchouli oil, 50g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing, add Oleum Arachidis hypogaeae semen to 250g, mixing is made 1000 of soft capsules (0.25g/ grain).Taking dose is: every day 2 times, each 2.
Example 7 (drop pill):
Get the 160g Macrogol 4000,70 ℃ of heating in water bath make fusion, add 0.5g Oleum Curcumae, 5g patchouli oil, 5g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing, and mixing splashes in the refrigerative simethicone, makes drop pill 1000 balls (0.18g/ ball).Taking dose is: every day 2 times, each 2 balls.
Example 8 (hard capsule):
Get 5g Oleum Curcumae, 20g patchouli oil, 20g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing,, get clathrate with 240g betacyclodextrin enclose; Add starch to 370g, mixing is made granule, and is dry below 60 ℃, sieves, and the capsule of packing into No. 1 is made 1000 of hard capsules (0.37g/ grain).Taking dose is: every day 2 times, each 2.
Example 9 (tablet):
Get 10g Oleum Curcumae, 8g patchouli oil, 25g Flos Chrysanthemi Indici carbon dioxide supercritical fluid extraction thing,, get clathrate, add starch to 250g with 240g betacyclodextrin enclose, mixing is made granule, and is dry below 60 ℃, be pressed into 1000 (0.25g/ sheets), the bag film-coat, promptly.Taking dose is: every day 2 times, each 2.
Below be the technique effect that the present invention of zoopery proof has:
One, interior resisting virus test
1, to the lethal protective effect of influenza virus infecting mouse
1.1 experiment material
Be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: ribavirin is provided lot number 080104 by the Hubei institute of Pharmaceutical Industry.
Laboratory animal: the NIH mice, single sex is provided by Guangdong Medical Lab Animal Center, SPF rank, body weight 13～15g.Mouse feed is the prescription that is rich in multiple composition.Feeding environment: 23 ± 2 ℃ of room temperatures, relative humidity 75 ± 10%.
Virus: Influenza A1 virus Mus lung adapted strain grinds the institute Viral Laboratory by Traditional Chinese Medicine University Of Guangzhou's heat and provides, and after mice enhancing virulence, goes down to posterity 2 times in chick embryo allantoic cavity.And measure its median lethal dose(LD 50) LD50=5.52, put in the frozen water and preserve.
1.2 experimental technique
Mice is divided into positive drug group (ribavirin), virus control group (being model group), present composition group, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group, 15 every group at random.Each is organized mice and slightly anaesthetizes with ether, selects 5 LD50 virus concentrations of Influenza A1 virus Mus lung adapted strain for use, collunarium infecting mouse, 4 every, about 0.05ml.1d begins the dosage gastric infusion by 0.16ml/10g before infect, every day 1 time, and 12d continuously, the viral infection matched group is irritated stomach with the equal-volume distilled water.Day by day observe animal disease symptom and death toll, observe 15d altogether.Calculate mortality rate, protective rate, on average survive natural law and increase in life span.
Dead protective rate=(viral infection matched group mortality rate-test group mortality rate)/viral infection matched group mortality rate;
Increase in life span=(test group on average survive natural law-viral infection matched group on average survive natural law/viral infection matched group natural law of on average surviving) * 100%.
1.3 experimental result
Behind the mouse infection influenza virus, big frequently-occurring disease is after 4d, and mice hair occurred alarmming, movablely reduces, loses weight, degradation symptom under the body temperature, death occurred since the 6th day.Flos Chrysanthemi Indici group, mice serious symptom, mortality rate are up to 100%, and to testing the 10th day, mice is all dead.All the other each treated animal sxs, a few symptoms is heavy, even dead.The results are shown in Table 1
Table 1 pair influenza virus causes the protective effect of dead mouse
| Group | Dosage mg/Kg | Death toll (only) | Survival number (only) | Mean survival time (d) | Mortality rate (%) | Dead protective rate (%) | Prolong vital rates (%) |
| The virus control group | ～～ | 13 | 2 | 7.07±3.59 | 86.67 | ||
| The ribavirin group | 10 | 2 | 13 | 14.07±2.46 ＊＊ | 13.33 | 84.62 ＊＊ | 99.06 |
| Present composition group | (2.7 Oleum Curcumae) | 9 | 6 | 11.27±4.18 ＊＊## | 60 | 30.77 | 45.28 |
| The Rhizoma Curcumae line of oils | 2.7 | 10 | 5 | 9.60±3.87 ＊＊##& | 66.67 | 23.08 | 35.79 |
| The Herba Pogostemonis line of oils | 13.3 | 12 | 3 | 9.26±3.37 ＊＊##& | 80 | 7.69 | 31.13 |
| The Flos Chrysanthemi Indici group | 13.3 | 15 | 0 | 5.40±2.67##&& | 100 | ～ | ～ |
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; Compare with ribavirin: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
1.4 discuss
The result shows, this drug regimen causes dead mouse to influenza virus protective effect is eager to excel than place an order private Oleum Curcumae, patchouli oil or Wild chrysanthemum extract of same concentrations.
2, to the influence of mice influenza virus property pneumonia
2.1 experiment material
Be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: ribavirin is provided lot number 080104 by the Hubei institute of Pharmaceutical Industry.
Laboratory animal: the NIH mice, single sex is provided by Guangdong Medical Lab Animal Center, SPF rank, body weight 13～15g.Mouse feed is the prescription that is rich in multiple composition.Feeding environment: 23 ± 2 ℃ of room temperatures, relative humidity 75 ± 10%.
Virus: Influenza A1 virus Mus lung adapted strain grinds the institute Viral Laboratory by Traditional Chinese Medicine University Of Guangzhou's heat and provides, and after mice enhancing virulence, goes down to posterity 2 times in chick embryo allantoic cavity.And measure its median lethal dose(LD 50) LD50=5.52, put in the frozen water and preserve.
2.2 experimental technique
Divide normal control group, positive drug group (ribavirin), virus control group (being model group), present composition group, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group at random with mice, except that the normal control group, 10 every group.Each is organized mice and slightly anaesthetizes with ether, selects 15 LD50 virus concentrations of Influenza A1 virus Mus lung adapted strain for use, collunarium infecting mouse, 4 every, about 0.05ml.1d begins the dosage gastric infusion by 0.16ml/10g before infect, every day 1 time, and 4d continuously, the viral infection matched group is irritated stomach with the equal-volume distilled water.Dissected after taking by weighing the mice body weight on the 6th day, the perusal pulmonary lesion, the degree of record pulmonary liver sample consolidation is won full lung and is weighed, and calculates the lung exponential quantity with body weight, and obtains lung index and suppression ratio.
Heavy (g) * 100%/mice body weight (g) of lung index=mouse lung
Lung index suppression ratio=(the average lung index of average lung index one experimental group of matched group) * average lung index of 100%/matched group
2.3 experimental result
Be condemned to death behind the mouse infection influenza virus 4d, the pulmonary lesion of dissecting discovery virus control group mice is obvious, and its lung index and normal control group relatively have significance difference (P＜0.01), show this model establishment.Ribavirin all has the obvious suppression effect for influenza virus induced mice pneumonia pathological changes, compares lung index take an evident turn for the better (P＜0.01) with the virus control group.Present composition group is compared with virus control, and the lung index is lower than virus control group (P＜0.05).The results are shown in Table 2.
The influence of the susceptible toxicity pneumonia of table 2 medicine convection current
| Grouping | Dosage mg/Kg | Lung index/x ± s | Suppression ratio/% |
| The normal control group | — | 0.81±0.14 | |
| The virus control group | ～～ | 1.52±0.17## | |
| The ribavirin matched group | 10.0 | 1.09±0.36 ＊＊ | 42.69 |
| Present composition group | (2.7 Oleum Curcumae) | 1.27±0.28 ＊ψ | 15.90 |
| The Rhizoma Curcumae line of oils | 2.7 | 1.35±0.28ψψ& | 12.92 |
| The Herba Pogostemonis line of oils | 13.5 | 1.38±0.25ψψ& | 9.42 |
| The Flos Chrysanthemi Indici group | 13.5 | 1.41±0.26ψψ& | 8.73 |
(compare ##:P＜0.01 with the normal control group; Compare with the virus control group:
*: P＜0.01,
*: P＜0.05; Compare with ribavirin: ψ P＜0.05, ψ ψ＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
2.4 discuss
The result shows, this drug regimen is eager to excel than place an order private Oleum Curcumae, patchouli oil or Wild chrysanthemum extract of same concentrations to the inhibitory action of mice influenza virus property pneumonia.
Two, extracorporeal antivirus effect test
1, material and method
1.1 cell: the human cervical carcinoma cell that goes down to posterity, Madin-Darby canine kidney(cell line) (MDCK) are provided by Inst. of Tropical Medicine, Guangzhou Chinese Medicine College.
1.2 virus: respiratory syncytial virus; Adenovirus type III; Coxsackie B 3 viruses, influenza virus A-prime Mus lung adapted strain FM1 is provided by Inst. of Tropical Medicine, Guangzhou Chinese Medicine College.
1.3 medicine and reagent drug regimen of the present invention, Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici extract are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Ribavirin injection (lot number 080104 is provided by the Hubei institute of Pharmaceutical Industry), IMDM cell culture fluid (GIBC0 company, the U.S.), newborn calf serum (Dalian biochemical reagents factory), trypsin SIGMA, the U.S.), tetrazolium bromide (MTT, SIGMA, the U.S.).
2, experimental technique
2.1 drug toxicity experiment
Respectively with the IMDM dilution that contains 2FBS, Oleum Curcumae dilutes 10 times of (27.31mg/ml), 100 times (2.73mg/ml), 200 times (1.36mg/ml), 400 times (0.68mg/ml), 600 times (0.46mg/ml), 800 times (0.34mg/ml), 1000 times (0.27mg/ml), 1200 times (0.23mg/ml), 1400 times (0.20mg/ml) with medicine to be measured.Drug regimen of the present invention is an experimental concentration with Oleum Curcumae concentration, and all the other patchouli oils, Flos Chrysanthemi Indici extract and ribavirin injection preparation are all the same.The human cervical carcinoma cell that will go down to posterity is inoculated in 96 well culture plates (0.2ml/ hole) by 2 * 108/ml, and culture fluid is fallen in cell monolayer hypsokinesis to be formed, adds above-mentioned 5 kinds of concentration medicine 0.2ml, and observation of cell pathological changes behind the 48h adopts mtt assay to measure the cell survival amount simultaneously.And adopt Reed-Muench method to calculate the maximal non-toxic concentration (TD0) and the median lethal concentration (TD50) of medicine.
2.2 the antiviral experiment in vitro of medicine
Experiment is divided into the normal cell matched group; The virus control group; Present composition group; The Rhizoma Curcumae line of oils; The Herba Pogostemonis line of oils; The Flos Chrysanthemi Indici extract group; Seven groups of ribavirin injection groups.By above-mentioned group technology, be inoculated in 96 well culture plates, normal control group observation of cell pathological changes, treat that the cytopathy of virus control group reaches ++ ++ the time, make the OD value to determine the cell survival amount with mtt assay mensuration, calculate the average suppression ratio (CPI) and the minimum inhibition dense (MIC) of each medicine pair cell pathological changes.Adopt Reed-Muench method calculating respectively to organize medicine inhibition virus and cause 50% cytopathic concentration (IC50), 90% valid density (IC90) and antiviral therapeutic index (TI).The TI value is the comprehensive judge index of medicine curative effect, and the toxicity of the big more explanation medicine of value is more little, and the valid density that suppresses virus is low more.
3, result
3.1 the medicine non-toxic of the experiment in vitro present composition and Oleum Curcumae TD0 as a result is 680 μ g/ml, TD50 is I360 μ g/ml.Get 680 μ g/ml, 340 μ g/ml, three concentration of 170 μ g/ml respectively for every group during experiment.Patchouli oil TD0 is I360 μ g/ml, and TD50 is 2730 μ g/ml.Flos Chrysanthemi Indici extract TD0 is 2730 μ g/ml, and TD50 is 27.31mg/ml.Contrast medicine ribavirin injection TD0 is 800 μ g/ml, and TD50 is I500 μ g/ml.
3.2 suppressing effect, the pathological changes of experiment in vitro medicine pair cell sees Table 3～table 6.In four groups of medicines, the CPI of present composition group and TI value are the highest in 4 kinds of antiviral experimental results, prove that the present composition all is better than independent Oleum Curcumae or patchouli oil to the inhibition effect of 4 kinds of viruses.
Table 3 different pharmaceutical causes cytopathic inhibition effect to respiratory syncytial virus
| Grouping | CPI (％) | MIC (μg/ml) | IC50 (μg/ml) | IC90 (μg/ml) | TI |
| Drug regimen of the present invention | 80.5 | 8 | 16 | 120 | 85 |
| Oleum Curcumae | 77 | 10 | 20 | 150 | 68 |
| Patchouli oil | 50 | 50 | 160 | 800 | 17 |
| Flos Chrysanthemi Indici extract | 12 | 1600 | 4500 | 20000 | 6.1 |
| Ribavirin | 70 | 25 | 50 | 200 | 30 |
Table 4 different pharmaceutical causes cytopathic inhibition effect to adenovirus Ad3 type
| Grouping | CPI (％) | MIC (μg/ml) | IC50 (μg/ml) | IC90 (μg/ml) | TI |
| Drug regimen of the present invention | 82.5 | 8 | 16 | 60 | 85 |
| Oleum Curcumae | 80 | 10 | 20 | 80 | 68 |
| Patchouli oil | 45 | 60 | 180 | 700 | 15.2 |
| Flos Chrysanthemi Indici extract | 10.5 | 1800 | 5000 | 21000 | 5.5 |
| Ribavirin | 70 | 25 | 50 | 200 | 30 |
Table 5 different pharmaceutical causes cytopathic inhibition effect to coxsackie B 3 type viruses
| Grouping | CPI (％) | MIC (μg/ml) | IC50 (μg/ml) | IC90 (μg/ml) | TI |
| Drug regimen of the present invention | 75 | 16 | 65 | 150 | 20.9 |
| Oleum Curcumae | 71 | 20 | 80 | 160 | 17 |
| Patchouli oil | 40 | 80 | 300 | 900 | 9.1 |
| Flos Chrysanthemi Indici extract | 11 | 1800 | 5500 | 23000 | 5 |
| Ribavirin | 70 | 25 | 50 | 200 | 30 |
Table 6 different pharmaceutical causes cytopathic inhibition effect to influenza virus A-prime Mus lung adapted strain FM1
| Grouping | CPI (％) | MIC (μg/ml) | IC50 (μg/ml) | IC90 (μg/ml) | TI |
| Drug regimen of the present invention | 75 | 16 | 65 | 150 | 20.9 |
| Oleum Curcumae | 74 | 20 | 45 | 160 | 30.2 |
| Patchouli oil | 35 | 90 | 320 | 900 | 8.5 |
| Flos Chrysanthemi Indici extract | 15 | 1500 | 4000 | 18000 | 6.8 |
| Ribavirin | 72 | 25 | 50 | 200 | 30 |
Three, antiinflammatory test (influence of xylol induced mice auricle edema)
1, experiment material
1.1 be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: prednisolone acetate, the 5mg/ sheet, Guangdong Huanan Pharmaceutical Co., Ltd produces, lot number: 070301.
1.2 laboratory animal: the NIH mice, the SPF level is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
2, experimental technique
Get the NIH mice, body weight 18～22g, 80, male and female half and half are divided into six groups at random, blank group, prednisolone acetate group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, patchouli oil, Flos Chrysanthemi Indici extract group.Except that the blank group, each group is pressed table 7 dosage gastric infusion respectively, and the blank group gives the isometric(al) distilled water, administration every day 1 time, continuous 4 days.Behind the last administration 0.5h, 100 μ l drip in the mouse right ear two sides with dimethylbenzene, left side ear is not coated with in contrast, put to death animal after causing scorching 0.5h, cut two ears, lay the same position of ears disk with diameter 9mm along the auricle baseline, electronic analytical balance, precision is weighed, with the difference of left and right auricle weight as the swelling degree, each group difference relatively.
3, experimental result and discussion
Experimental result sees Table 7
Table 7 respectively organize medicine xylol induced mice auricle edema influence (x ± s, n=10)
| Group | Dosage (the mg medicine/kg) | Ear swelling degree (mg) |
| The blank group | — | 13.26±1.17 |
| Prednisolone acetate | 40.0 | 4.71±1.12 ＊＊ |
| The present composition | (2.7 Oleum Curcumae) | 6.42±1.78 ＊＊ |
| The Rhizoma Curcumae line of oils | 2.7 | 9.11±1.31 ＊＊& |
| The Herba Pogostemonis line of oils | 13.5 | 8.07±1.62 ＊＊& |
| The Flos Chrysanthemi Indici extract group | 13.5 | 7.63±1.24 ＊＊& |
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; With prednisolone acetate: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
Table 7 is the result show, compare with the blank group, prednisolone acetate group, present composition group, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group all can significantly suppress the ear swelling degree (P value equal＜0.01) of the scorching mice of caused by dimethylbenzene xylene, point out each experimental group that antiinflammatory action is preferably arranged.Compare with the prednisolone acetate group, present composition group, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group, the P value is equal〉0.05, point out drug regimen antiinflammatory action intensity of the present invention suitable, and its antiinflammatory action is stronger than using Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici extract separately with prednisolone acetate.
Four, separate heat test
1, experiment material
1.1 be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: etidocine tablet (indometacin enteric-coated tablet), the 5mg/ sheet, Guangdong Huanan Pharmaceutical Co., Ltd produces, lot number: 071001.
1.2 laboratory animal: the SD rat, the SPF level is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
2., experimental technique
Get some of rats, body weight (200 ± 20) g, choose 50 of the rat of body temperature in 36.6 ℃～38.3 ℃, male and female half and half, be divided into 5 groups at random, 10 every group, gastric infusion, administration volume 20ml/kg, successive administration 3, fasting 12 before experiment is freely drunk water, the same day was surveyed body temperature 2 times in experiment, every 2h 1 time, select the temperature difference to be no more than 0.3 ℃ 80 of qualified rats, after the administration at once in rat back subcutaneous injection dry yeast (15) suspension 10ml/kg, then in 2,4,6,10h measures anus temperature value, comparison between organizing.
3, experimental result and discussion
Experimental result sees Table 8
Table 8 respectively organize medicine to fresh yeast cause rat fever influence (x ± s, n=10)
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; Compare with indometacin: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
Table 8 is the result show, compare with the blank group, indometacin has refrigeration function to the heating that fresh yeast causes rat behind administration 2～4h, drug regimen of the present invention after administration 3,4h has refrigeration function, the Flos Chrysanthemi Indici extract group has refrigeration function during 4h after administration, all the other each groups all do not have refrigeration function.
Five, to Immune Effects
1, experiment material
1.1 be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: prednisolone acetate, the 5mg/ sheet, Guangdong Huanan Pharmaceutical Co., Ltd produces, lot number: 070301.
1.2 laboratory animal: the NIH mice, the SPF level is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
2, experimental technique
Get 60 of NIH mices, male and female half and half, body weight are 18～22g, are divided into six groups at random, blank group, prednisolone acetate (immunosuppressant) group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group.Except that the blank group, each group is pressed table 9 dosage gastric infusion respectively, and the blank group gives the isometric(al) distilled water, every day 1 time, 7d continuously.After the last administration 0.5 hour, inject the Chinese prepared Chinese ink of dilution (with 4 times of 1% gelatin solution dilutions) 0.1mL/10g in the tail vein, 2min and 12min after injecting prepared Chinese ink, get blood 10 μ L with special suction pipe from the mouse orbit rear vein beard, be blown at once among the 0.1% Na2CO3 solution 10mL, suck from this liquid with suction pipe and to blow out for several times, fully wash out the blood that the suction pipe wall adheres to, be dissolved in 10mL0.1% Na2CO3 solution school zero with 0.025mL normal mouse blood, measure trap (A) value in the 576nm place, calculate phagocytic index K, experimental data adopts relatively each group difference of one factor analysis of variance.
K＝(logOD1～logOD2)/(t1～t2)
3, experimental result and discussion
Table 9 respectively organize medicine to the influence of mice reticuloendothelial system phagocytic function (x ± s, n=10)
| Group | Dosage (mg/kg) | Phagocytic index K |
| Blank | — | 0.006±0.002 |
| Prednisolone acetate | 40.0 | 0.003±0.001 ＊＊ |
| Drug regimen of the present invention | (2.7 Oleum Curcumae) | 0.018±0.004 ＊＊ |
| Oleum Curcumae | 2.7 | 0.017±0.002 ＊＊ |
| Patchouli oil | 13.5 | 0.014±0.003 ＊＊ |
| Flos Chrysanthemi Indici extract | 13.5 | 0.010±0.002&& |
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; Compare with prednisolone acetate: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
Table 9 is the result show, compare with the blank group, drug regimen of the present invention, Oleum Curcumae, patchouli oil, all can significantly improve the phagocytic function (P＜0.01) of mice reticuloendothelial system, the prednisolone acetate group can significantly reduce the phagocytic function (P＜0.01) of mice reticuloendothelial system, the effect of pointing out drug regimen of the present invention to be significantly improved immunologic function.
Six, analgesic experiment (to the influence of the mice hot plate threshold of pain)
1, experiment material
1.1 be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: etidocine tablet (indometacin enteric-coated tablet), the 5mg/ sheet, Guangdong Huanan Pharmaceutical Co., Ltd produces, lot number: 071001.
1.2 laboratory animal: the NIH mice, male, the SPF level is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
2, experimental technique
Regulating thermostat makes water temperature be controlled at (55 ± 0.5) ℃, hot plate preheating 30min.Survey the normal pain threshold of mice: get 18～22g female mice number, each one is placed on the hot plate, write down its begin to lick metapedes required time (second) for this reason the normal pain threshold of Mus (less than 5 seconds or greater than 30 seconds or middle leaper need not).Qualified mice is divided into 6 groups at random, measures its pain threshold and be pain threshold before its administration.Be divided into blank group, indometacin group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group, except that the blank group, each group is pressed table 10 dosage gastric infusion respectively, and the blank group gives the isometric(al) distilled water, administration every day 1 time, 4d continuously.Behind the last administration 60min, measure the mice pain threshold respectively.If 60 seconds reactionless, its pain threshold calculated by 60 seconds.Experimental data adopts relatively each group difference of one factor analysis of variance.
3, experimental result and discussion
Experimental result sees Table 10
Table 10 respectively organize medicine to the influence of mice hot plate method pain threshold (x ± s, n=10)
| Group | Dosage (mg/kg) | The preceding threshold of pain of medicine (s) | The threshold of pain behind the medicine (s) |
| Blank | 13.34±3.67 | 14.29±3.81 | |
| Etidocine tablet | 1.67 | 12.75±3.65 | 23.07±4.35 ＊＊ |
| Drug regimen of the present invention | (2.7 Oleum Curcumae) | 13.56±4.53 | 25.54±3.19 ＊＊ |
| Oleum Curcumae | 2.7 | 12.92±3.18 | 19.65±4.07 ＊ |
| Patchouli oil | 13.5 | 12.83±4.22 | 18.43±4.25 ＊ |
| Flos Chrysanthemi Indici extract | 13.5 | 13.01±3.58 | 16.82±3.96## |
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; Compare with indometacin: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
The result shows, with the comparison of blank group, indometacin group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, all can suppress the pain of hot plate induced mice, points out above-mentioned each experimental group that analgesic activity is preferably all arranged.Compare with the indometacin group, drug regimen group zero difference of the present invention, the analgesic activity intensity of prompting drug regimen of the present invention is equivalent to indometacin.
Seven, calm test (to the influence test of mice autonomic activities number of times)
1, experiment material
1.1 be subjected to the reagent thing: drug regimen of the present invention (containing Oleum Curcumae 2.7g, patchouli oil 13.5g, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing 13.5g), Oleum Curcumae, patchouli oil, Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing are provided by Traditional Chinese Medicine University Of Guangzhou new drug development research center; Positive control drug: aspirin tablet, the 20mg/ sheet, Guangzhou BaiYunshan GuangHua Pharmacy Co., Ltd produces, lot number: 070201.
1.2 laboratory animal: the NIH mice, male and female half and half, the SPF level is provided by Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center.
2, experimental technique
Get 50 of NIH mices, male and female half and half, body weight 18～22 is divided into six groups at random, 10 every group, is blank group, aspirin group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils, Flos Chrysanthemi Indici extract group.Except that the blank group, each group is pressed table 11 dosage gastric infusion respectively, and the blank group gives the isometric(al) distilled water, administration every day 1 time, 4d continuously.Behind last administration 1h, the 2h, mice is put in the mice autonomic activities monitor, make it adapt to 3min after, movable number of times in the record 5min, adopt the variance analysis of repeated measure organize between relatively.
3, experimental result and discussion
Experimental result sees Table 11
Table 11 respectively organize medicine to the influence of mice autonomic activities (x ± s, n=10)
| Group | Dosage (mg/kg) | Movable number of times in the 5min behind the administration 1h/time | Movable number of times in the 5min behind the administration 2h/time |
| Blank | 127.2±18.1 | 123.1±15.8 | |
| Aspirin | 78 | 61.3±13.2 ＊＊ | 53.2±18.6 ＊＊ |
| Medicine group of the present invention | (2.7 Oleum Curcumae) | 87.0±29.3 ＊＊# | 78.5±26.3 ＊＊# |
| Oleum Curcumae | 2.7 | 84.5±34.6 ＊＊# | 92.8±27.8 ＊＊## |
| Patchouli oil | 13.5 | 86.5±31.2 ＊＊# | 96.0±25.1 ＊＊## |
| Flos Chrysanthemi Indici extract | 13.5 | 82.6±25.1 ＊＊# | 85.0±30.5 ＊＊# |
(compare with the virus control group:
*P＜0.05,
*: P＜0.01; Compare with aspirin: #P＜0.05, ##＜0.01; Compare: ﹠amp with drug regimen of the present invention;＜0.05 ， ﹠amp; ﹠amp; : P＜0.01)
The result shows, compare with the blank group, aspirin group, drug regimen group of the present invention, Rhizoma Curcumae line of oils, Herba Pogostemonis line of oils all can significantly reduce the autonomic activities number of times (the P value all＜0.01) in the mice 5min, point out above-mentioned each experimental group that sedation is preferably all arranged.
Claims (6)
1, a kind of anti-drug regimen that cures cold is characterized in that being made up of following materials of weight proportions medicine: 0.5～20 part of Oleum Curcumae, 5～50 parts of patchouli oils, 5～50 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things.
2, the anti-drug regimen that cures cold according to claim 1 is characterized in that each composition weight proportion is preferably: 2.7 parts of Oleum Curcumae, 13.5 parts of patchouli oils, 13.5 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things.
3, the anti-drug regimen that cures cold according to claim 1 is characterized in that: the application of described pharmaceutical composition aspect respiratory system disease is used to catch a cold, the prevention and the treatment of influenza and other viral infections.
4, the anti-drug regimen that cures cold according to claim 1, it is characterized in that: described this pharmaceutical composition and extract or refining thing can add conventional adjuvant or excipient, make clinical acceptable forms, are peroral dosage form.
5, the anti-drug regimen that cures cold according to claim 4 is characterized in that: described peroral dosage form is selected from a kind of in the middle of pill, tablet, capsule, spray and the drop pill.
6, a kind of preparation method of the anti-drug regimen that cures cold is characterized in that, comprises the steps:
1., at first prepare Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing, get the Flos Chrysanthemi Indici crude drug, be ground into coarse powder, put in the supercritical carbon dioxide extraction instrument and extract, the extraction parameter is 45 ± 3 ℃ of extraction temperature, extracting pressure 20 ± 10MPa, parsing pressure is 6～7MPa, 55 ± 5 ℃ of resolution temperatures, carbon dioxide flow is 25 ± 3L/h, the extraction time is 2 ± 1h, and the gained extract is put in the clean container, and is standby;
2., again with the Flos Chrysanthemi Indici supercritical carbon dioxide extraction thing of above-mentioned preparation, respectively become weight proportion by drug regimen: 0.5～20 part of Oleum Curcumae, 5～50 parts of patchouli oils, 5～50 parts of Flos Chrysanthemi Indici supercritical carbon dioxide extraction things, above material mixing stirs, and makes drug regimen then.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010096987A1 (en) * | 2009-02-27 | 2010-09-02 | 东莞广州中医药大学中医药数理工程研究院 | Use of patchouli alcohol in medicament preparation |
| CN102895218A (en) * | 2012-10-29 | 2013-01-30 | 成都中医药大学 | Novel application of patchouli alcohol |
| CN103191183A (en) * | 2012-07-30 | 2013-07-10 | 成都中医药大学 | Use of palchouli oil in preparation of antiviral drug |
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| CN1326552C (en) * | 2004-07-22 | 2007-07-18 | 广州中医药大学科技产业园有限公司 | Medicine for treating viral influenza, and its prepn. method |
| CN100493532C (en) * | 2006-01-19 | 2009-06-03 | 广州星群（药业）股份有限公司 | Traditional Chinese medicine effective part for treating influenza virus and its preparation process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2010096987A1 (en) * | 2009-02-27 | 2010-09-02 | 东莞广州中医药大学中医药数理工程研究院 | Use of patchouli alcohol in medicament preparation |
| CN103191183A (en) * | 2012-07-30 | 2013-07-10 | 成都中医药大学 | Use of palchouli oil in preparation of antiviral drug |
| CN103393752A (en) * | 2012-07-30 | 2013-11-20 | 成都中医药大学 | New application of palchouli oil |
| CN102895218A (en) * | 2012-10-29 | 2013-01-30 | 成都中医药大学 | Novel application of patchouli alcohol |
| CN110742924A (en) * | 2019-11-04 | 2020-02-04 | 陈志维 | Traditional Chinese medicine volatile oil composition and application thereof |
| CN110742924B (en) * | 2019-11-04 | 2021-07-16 | 陈志维 | Traditional Chinese medicine volatile oil composition and application thereof |
| CN110859900A (en) * | 2019-12-20 | 2020-03-06 | 雷允上药业集团有限公司 | Application of liuling detoxification pills in preparation of medicines for preventing and treating cold diseases |
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