CN103120721A - Chinese medicinal composition for treating wind-heat common cold and preparation method and application thereof - Google Patents
Chinese medicinal composition for treating wind-heat common cold and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a Chinese medicinal composition for treating wind-heat common cold and a preparation method thereof. The Chinese medicinal composition comprises the following raw medicaments: 400 to 800 parts by weight of honeysuckle, 400 to 800 parts by weight of trollflower, 100 to 300 parts by weight of gentiana dahurica fisch flower, and 700 to 1,300 parts by weight of isatis root. The preparation method comprises the following steps of: acquiring the raw medicaments in ratio and extracting the raw medicaments by adopting an organic solvent. The invention also provides application of the Chinese medicinal composition in a medicament for treating the wind-heat common cold and in preparation of an influenza virus inhibiting medicament and an anti-inflammation medicament. The medicinal composition is prepared from the honeysuckle, the trollflower, the gentiana dahurica fisch flower and the isatis root, or prepared from a honeysuckle extract, a trollflower extract, a gentiana dahurica fisch flower extract and an isatis root extract. The Chinese medicinal composition has obvious effects of resisting viruses, clearing heat, relieving pain and diminishing the inflammation.
Description
Technical field
The present invention relates to a kind of Chinese medicine composition and preparation method thereof, be specifically related to a kind of Chinese medicine composition for the treatment of anemopyretic cold and preparation method thereof.
Background technology
Flos Lonicerae is the dry flower of caprifoliaceae plant Radix Ophiopogonis Lonicera japonica Thunb. or the flower that band is just opened.Its chemical composition mainly includes machine acids, flavonoid, triterpene saponin and volatilization wet goods.The chlorogenic acid compound is the main effective ingredient of Flos Lonicerae, comprises chlorogenic acid (chlorogenic acid) and isochlorogenic acid (isochlorogenic acid), and other organic acid also has caffeic acid and Palmic acid.Pharmacological evaluation shows that Flos Lonicerae all has certain inhibitory action to various pathogens.
Flos Trollii Trolliusch inensis Bunge is the Ranunculaceae herbaceos perennial, is used as medicine with flower, and heat-clearing and toxic substances removing, antibacterial and anti-inflammation functions are arranged.Modern age, pharmacological research showed, Flos Trollii all has obvious inhibitory action to bacillus pyocyaneus, dysentery bacterium, staphylococcus aureus etc., and the death that influenza virus infected causes has significant protective effect.
Gentiana dahurica flower is the dried floral of gentianaceae plant Gentiana dahurica Gentiana dahurica Fisch., and the effect of have heat clearing away, detoxifcation, cough-relieving, eliminating the phlegm for cough due to lung-heat, laryngopharynx swelling and pain, scorchingly hot, distemper etc., is Mongolian medicine DANFU side common medicine.Its chemical constitution study shows that it may contain flavonoid and glycosides thereof, triterpene saponin, steroidal saponin, alkaloid, organic acid, tannin, saccharide, protein etc.
Radix Isatidis is the dry root of cruciferae isatis Isatis indigotica Fort., and heat-clearing and toxic substances removing is arranged, removing heat from blood sore-throat relieving function.The Radix Isatidis that studies show that of chemical composition contains alkaloid (indigo class indole alkaloid, quinazolone Alkaloid), organic acid compound, flavonoid, Semen Sinapis glycoside, sterols, polysaccharide and aminoacid etc. at present.Pharmacological research shows, Radix Isatidis has obvious inhibitory action to influenza A virus.
Summary of the invention
The object of the present invention is to provide a kind of Chinese medicine composition for the treatment of flu;
Another object of the present invention is to provide the preparation method of this Chinese medicine composition;
Another object of the present invention is to provide the application of this Chinese medicine composition in preparation treatment anemopyretic cold medicine;
Another object of the present invention is to provide the application of this Chinese medicine composition in preparation inhibition influenza virus property medicine;
Another object of the present invention is to provide the application of this Chinese medicine composition in the preparation anti-inflammatory drug.
The objective of the invention is to be achieved through the following technical solutions:
A kind of Chinese medicine composition for the treatment of anemopyretic cold, the crude drug of this Chinese medicine composition consists of:
Flos Lonicerae 400-800 weight portion Flos Trollii 400-800 weight portion
Gentiana dahurica flower 100-300 weight portion Radix Isatidis 700-1300 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae 500-700 weight portion Flos Trollii 500-700 weight portion
Gentiana dahurica flower 150-250 weight portion Radix Isatidis 800-1200 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae 550-650 weight portion Flos Trollii 550-650 weight portion
Gentiana dahurica flower 180-220 weight portion Radix Isatidis 900-1100 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae 600 weight portion Flos Trollii 600 weight portions
Gentiana dahurica is spent 200 weight portion Radix Isatidis 1000 weight portions.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae 550 weight portion Flos Trollii 650 weight portions
Gentiana dahurica is spent 180 weight portion Radix Isatidis 1100 weight portions.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae 650 weight portion Flos Trollii 550 weight portions
Gentiana dahurica is spent 220 weight portion Radix Isatidis 900 weight portions.
The preparation method of Chinese medicine composition of the present invention is: get in proportion crude drug, adopt organic solvent extraction.
Preferably, described organic solvent is ethanol or methanol; Further preferred, described organic solvent is the ethanol of 50-90%;
Preferably, extracting solvent load is that 5-10 doubly measures (volume/weight); Extraction time is 2-4 time, each 1-3 hour; Further preferred, extracting solvent load is 7 times of amounts (volume/weight); Extraction time is 3 times, each 1 hour.
Chinese medicine composition of the present invention can also prepare as follows: get in proportion crude drug, adopt the decoction and alcohol sedimentation technique preparation; Wherein said alcohol precipitation concentration is 40-90%.
Preferably, described alcohol precipitation concentration is 60%;
Preferably, described extraction solvent load is that 5-10 doubly measures (volume/weight); Extraction time is 2-4 time, each 1-3 hour; Further preferred, extracting solvent load is 6 times of amounts (volume/weight); Extraction time is 3 times, each 1 hour.
Chinese medicine composition of the present invention can also prepare as follows: get in proportion crude drug, wherein Flos Lonicerae, Flos Trollii, Gentiana dahurica flower merge employing organic solvent united extraction, get extract A; Radix Isatidis adopts separately decoction and alcohol sedimentation technique to be prepared into extract B; United extraction thing A and B, and get final product.
Wherein said alcohol precipitation concentration is 40-90%, is preferably 60%;
Described organic solvent is the ethanol of 50-90%; Described extraction solvent load is that 5-10 doubly measures (volume/weight); Extraction time is 2-4 time, each 1-3 hour; Further preferred, extracting solvent load is 6 times of amounts (volume/weight); Extraction time is 3 times, each 1 hour.
Chinese medicine composition of the present invention clinical acceptable dosage forms such as the agent of technique granulation, tablet, powder, capsule, pill, oral liquid routinely after extracting.
For above-mentioned dosage form can be realized, need to add the acceptable adjuvant of pharmacy when these dosage forms of preparation, such as: filler, disintegrating agent, lubricant, suspending agent, binding agent, sweeting agent, correctives, antiseptic, substrate etc.Filler comprises: starch, pregelatinized Starch, lactose, mannitol, chitin, microcrystalline Cellulose, sucrose etc.; Disintegrating agent comprises: starch, pregelatinized Starch, microcrystalline Cellulose, carboxymethyl starch sodium, crospolyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose etc.; Lubricant comprises: magnesium stearate, sodium lauryl sulphate, Pulvis Talci, silicon dioxide etc.; Suspending agent comprises: polyvinylpyrrolidone, microcrystalline Cellulose, sucrose, agar, hydroxypropyl emthylcellulose etc.; Binding agent comprises, starch slurry, polyvinylpyrrolidone, hydroxypropyl emthylcellulose etc.; Sweeting agent comprises: saccharin sodium, Aspartane, sucrose, cyclamate, enoxolone etc.; Correctives comprises: sweeting agent and various essence; Antiseptic comprises: parabens, benzoic acid, sodium benzoate, sorbic acid and its esters, benzalkonium bromide, acetic acid chloroethene are decided, the Folium eucalypti globueli (Eucalyptus globulus Labill.) wet goods; Substrate comprises: PEG6000, PEG4000, insect wax etc.
The Chinese medicine composition for the treatment of anemopyretic cold of the present invention is except the form that feeds intake with Flos Lonicerae, Flos Trollii, Gentiana dahurica flower and Radix Isatidis crude drug, therefore the form that can also adopt the extract (effective site) with Flos Lonicerae, Flos Trollii, Gentiana dahurica flower and Radix Isatidis to feed intake the present invention further discloses the Chinese medicine composition for the treatment of anemopyretic cold:
A kind of Chinese medicine composition for the treatment of anemopyretic cold, the crude drug of this Chinese medicine composition consists of:
Flos Lonicerae extract 400-800 weight portion Flos Trollii extract 400-800 weight portion
Gentiana dahurica flower extract 100-300 weight portion Radix Isatidis extract 700-1300 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae extract 500-700 weight portion Flos Trollii extract 500-700 weight portion
Gentiana dahurica flower extract 150-250 weight portion Radix Isatidis extract 800-1200 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae extract 550-650 weight portion Flos Trollii extract 550-650 weight portion
Gentiana dahurica flower extract 180-220 weight portion Radix Isatidis extract 900-1100 weight portion.
Chinese medicine composition of the present invention, its crude drug composition is preferably:
Flos Lonicerae extract 600 weight portion Flos Trollii extract 600 weight portions
Gentiana dahurica flower extract 200 weight portion Radix Isatidis extract 1000 weight portions.
Flos Lonicerae extract of the present invention, Flos Trollii extract, Gentiana dahurica flower extract, Radix Isatidis extract can be but be not limited to ethanol extraction or water extract-alcohol precipitation extract; Ethanol extraction or water extract-alcohol precipitation extract can also further be made with extra care purification, as crossing macroporous resin column.
Described extraction concentration of alcohol is 50-90%, and solvent load is that 5-10 doubly measures (volume/weight); Extraction time is 2-4 time, each 1-3 hour;
Described aqueous extraction-alcohol precipitation technology, extracting solvent load is that 5-10 doubly measures (volume/weight); Extraction time is 2-4 time, each 1-3 hour; Alcohol precipitation concentration is 40-90%.
Decoction and alcohol sedimentation technique of the present invention means in the Chinese medicine water extracting liquid, adds ethanol to make to reach the alcohol amount (being alcohol precipitation concentration) that contains of regulation, and some composition dissolubility in alcoholic solution reduces separates out precipitation, filter, get alcoholic solution, reclaim solvent, the method that water extraction liquid is made with extra care.
The present composition has obvious antiviral, analgesic, analgesic and anti-inflammatory effects.The viral pneumonia suppression ratio is reached 72.3%, significantly be better than model group (P＜0.05), and the life span of energy significant prolongation influenza infection mice; Yeast is caused the rat temperature rising also have obvious refrigeration function, its effect is better than Chinese medicine positive control drug SHUANGHUANGLIAN KOUFUYE; Antalgic and inflammation relieving experiment shows the stomachache writhing response (P＜0.01) that the present composition can obviously suppress lumbar injection acetic acid and causes, and can significantly alleviate the swelling of mice toes.
Chinese medicine composition of the present invention consumption clinically is 36g crude drug/people/sky, and people's body weight is pressed 60kg and calculated, and clinical dosage is equivalent to 0.6g crude drug/kg.Be designed to three dosage levels in animal experiment, be about respectively 1/2 clinical amount, clinical amount, 2 times of clinical amounts.Press body surface area conversion, rat dosage be respectively 1.75,3.5,7.0g crude drug/kg, mice dosage is respectively 2.5,5.0,10.0g crude drug/kg, each approximately is equivalent to 0.5,1.0,2.0 times of clinical dosage.In each test, all animals are all adopted gastric infusion.
Experimental example
Experimental example 1 antiviral experiment
1 experiment material
1.1 Experimental agents:
Present composition granule (pressing embodiment 1 method preparation) is mixed with 0.5g crude drug/ml with distilled water with it;
Oseltamivir phosphate capsule (PVG Ying Xuan company) is mixed with 2.5mg/ml with distilled water with it;
Antivirus oral liquid (Noon pharmacy Co., Ltd., Hubei Prov) is mixed with 0.25ml stock solution/ml with distilled water with it.
1.2 Strain: FM1 type mice strains of influenza viruses is provided by pharmacological room of Institute Of Chinese Materia Medica Of China Academy of Chinese Medical Sciences virus group
1.3 laboratory animal: the ICR mice, available from Beijing Vital River Experimental Animals Technology Co., Ltd., licence is numbered SCXK (capital) 2006-0009
2 experimental techniques
2.1 the impact of present composition influenza virus infected lung index
Selective body focuses on the healthy ICR mice between 13-15g, and 100, male and female half and half are divided into animal at random: 1. Normal group, 2. model group, 3. oseltamivir phosphate capsule 50mg/kg, 4. antivirus oral liquid 5ml/kg, 5. the present composition 10.0 crude drugs/kg is totally 5 groups, and 20 every group, each 10 of male and female.
The all beginning administrations in 1 day before the zoogenetic infection influenza virus of each group, the administration volume is the 0.2ml/10g body weight, every day, gastric infusion was 1 time.Next day, except Normal group, the animal of all the other groups is slightly anaesthetized with ether, with 15 times of LD
50Influenza virus drop nose infecting mouse, every animal collunarium volume is 35 μ l, infects to cause viral pneumonia.Infect and continue administration on the same day and 3 days subsequently, successive administration is 5 days altogether.Normal group and model group gavage give with the volume distilled water.The 4th administration began fasting afternoon on the same day, and fasting 17～20h freely drinks water.Claim the fasting body weight, 1h after last 1 administration, all animals are plucked and put to death after eyeball is got blood, and weighing lungs weight is calculated lung exponential sum pneumonia suppression ratio (%).
Lung index (PI)=lungs weight (g)/body weight (g) * 100
Pneumonia suppression ratio (%)=(model group PI-medicine group PI)/(model group PI-matched group PI) * 100
2.2 the impact of TNF-alpha content in present composition influenza virus infected lung homogenate
Randomly draw 11 animals (6 of ♂ from 2.1,5 of ♀) carry out lung tissue homogenate, be prepared into 10% homogenate with normal saline, 3000 leave heart 15min, supernatant detects the content of TNF-α with the ELISA test kit, the BCA method is measured protein content, and calculates the content of TNF-α in every milligram of albumen.
2.3 the impact of present composition influenza virus infected survival day
Animal grouping, administration are with 2.1, and the animal survival situation is observed in successive administration drug withdrawal after 5 days day by day, observes altogether 15 days.Record the animal survival natural law and calculate rate elongation.
Rate elongation (%)=(survival day medicine group-survival day model group)/survival day model group * 100
3 experimental results
3.1 the impact of present composition influenza virus infected lung index
The results are shown in Table 1.
The impact of table 1 present composition influenza virus infected lung index
Annotate: compare * P＜0.05 with model group; * P＜0.01
The demonstration of table 1 result, model group is due to pulmonary infection, and there are obvious inflammation and edema in pulmonary, and the weight of lung obviously increases (comparing P＜0.01 with matched group) than matched group.The present composition has obvious inhibitory action to viral pneumonia, has compared statistical significant difference (P＜0.05) with model group, and the pneumonia suppression ratio reaches 72.3%.Result shows that the present composition has the effect of significant inhibition viral pneumonia.
3.2 the impact of TNF-alpha content in present composition influenza virus infected lung homogenate
The results are shown in Table 2.
Table 2 present composition on the impact of TNF-alpha content in the homogenate of Pneumonia Mice lung tissue (
Pg/mg pro, n=11)
Annotate: compare * * P＜0.01 with model group
Result show positive control drug oseltamivir phosphate capsule and antivirus oral liquid to the secretion of TNF-α in lung tissue homogenate without reducing effect, and the present composition can reduce the content (comparing P＜0.01 with model group) of TNF-α to a certain extent, thereby prevents increasing the weight of of pneumonia reaction.
3.3 the impact of present composition influenza virus infected survival day
The results are shown in Table 3.
The impact of table 3 present composition influenza virus infected survival day
| Group | Dosage (g/kg) | Survival day (my god) | Rate elongation (%) |
| Contrast | - | 15.0±0.0*** | - |
| Model | - | 11.4±3.7 | - |
| Oseltamivir phosphate capsule | 0.05 | 15.0±0.0*** | 31.6 |
| Antivirus oral liquid | 5.0ml/kg | 14.2±1.7** | 24.6 |
| The present composition | 10.0 | 13.2±3.0* | 15.8 |
Annotate: compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with model group
Result shows, the life span of model group is 11.3 days, and present composition group makes death time of animal extend to 13.2 days, and rate elongation is 15.8%, with model group, significant (P＜0.05) is arranged relatively.Show that present composition infected by influenza infects the dead mouse that causes and has protective effect.
The analgesic experiment of experimental example 2
1 experiment material
1.1 Experimental agents:
Present composition granule (pressing embodiment 1 method preparation) is mixed with 0.7g crude drug/ml with distilled water with it, 0.35g crude drug/ml;
Aspirin effervescent tablets (AstraZeneca pharmaceutical Co. Ltd) is mixed with 20mg/ml with distilled water with it;
SHUANGHUANGLIAN KOUFUYE (Harbin Pharmaceutical Group, Sanjing Pharmaceutical Co., Ltd.) is mixed with 0.6ml stock solution/ml with distilled water with it.
1.2 laboratory animal: the SD rat, available from Beijing Vital River Experimental Animals Technology Co., Ltd., licence is numbered SCXK (capital) 2006-0009
2 experimental techniques
Rat is divided into normal control treated animal and modeling animal at random by body weight.Except 10 of normal control treated animals, all the other 60 equal modelings.Before modeling, measured the basal body temperature of animal, and got the anus temperature meansigma methods of twice mensuration as its basal body temperature in continuous 2 days.All according to the volume subcutaneous injection 15% yeast suspension of 2ml/100g body weight, the normal control treated animal is injected the equal-volume normal saline to the modeling animal.3.5h measures rat temperature after the injection yeast.Animal after modeling is divided into 6 groups at random according to the body temperature increased value, is respectively: 1. model group, 2. aspirin 200mg/kg group, 3. SHUANGHUANGLIAN KOUFUYE 6ml/kg organizes, 4. present composition 3.5g crude drug/kg organizes, and 5. present composition 7.0g crude drug/kg organizes, every group of 10 animals.Each organizes the equal gastric infusion of rat, administration volume 1ml/100g.0.5h, 1.0h, 2.0h, 3.0h, 4.0h, 6.0h measure respectively rat anus temperature after administration.Each group of 3.0h is strengthened administration 1 time again after the 1st administration.Normal group and model group gavage equal-volume distilled water.The difference of calculating basal body temperature before each time point of every animal and pyrogenicity is as body temperature increased value (body temperature-basal body temperature of certain time point after Δ ℃=pyrogenicity), each is organized body temperature increased value data and all represents with mean ± standard deviation, between organizing with one factor analysis of variance (ANOVA) relatively.Calculate its suppression ratio % by following formula.
Suppression ratio (%)=(body temperature increased value model group-body temperature increased value medicine group)/body temperature increased value model group * 100%
3 experimental results
The results are shown in Table 4.
Table 4 present composition on yeast cause the rat fever refrigeration function impact (
N=10)
Annotate: compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with model group
Table 4 result shows, the present composition causes the rat temperature rising to yeast and also has obvious refrigeration function, 3.5g crude drug/kg dosage group 0.5h and 1.0h after medicine, 7.0g crude drug/kg dosage group 6.0h after medicine all can obviously suppress the rising of rat temperature, suppression ratio is respectively 18.8%, 25.7% and 33.5%, with model group, significant difference (P＜0.05, P＜0.01, P＜0.01) is arranged relatively.Western medicine positive drug aspirin group 0.5h to 6h after medicine all can obviously suppress the fervescence (P＜0.001) of rat; Chinese medicine positive control drug SHUANGHUANGLIAN KOUFUYE refrigeration function in this experiment is not obvious.
Result shows that the high heat of rat that the present composition is induced yeast has certain refrigeration function, and its effect is better than Chinese medicine positive control drug SHUANGHUANGLIAN KOUFUYE.
Experimental example 3 antiinflammatory experiments
1 experiment material
1.1 Experimental agents:
Present composition granule (pressing embodiment 1 method preparation) is mixed with 4 kinds of concentration with distilled water with it, is respectively 0.5g crude drug/ml, 0.25g crude drug/ml (mice) and 0.7g crude drug/ml, 0.35g crude drug/ml (rat);
Aspirin effervescent tablets (AstraZeneca pharmaceutical Co. Ltd) is mixed with 10mg/ml (rat) and 20mg/ml (mice) with distilled water with it;
SHUANGHUANGLIAN KOUFUYE (Harbin Pharmaceutical Group, Sanjing Pharmaceutical Co., Ltd.) is mixed with 0.6ml stock solution/ml (rat) and 0.9ml stock solution/ml (mice) with distilled water with it.
1.2 laboratory animal: the ICR mice, the SD rat, all available from Beijing Vital River Experimental Animals Technology Co., Ltd., licence is numbered SCXK (capital) 2006-0009
2 experimental techniques
2.1 the impact of the present composition on the mice capillary permeability
The ICR mice, male, 76.Be divided at random by body weight: 1. Normal group, 2. model group, 3. aspirin 200mg/kg, 4. SHUANGHUANGLIAN KOUFUYE 9ml/kg, 5. present composition 2.5g crude drug/kg, 6. present composition 5.0g crude drug/kg is totally 6 groups, and every group of 12-13 is only.Each group gives medicine 0.2ml/10g by the body weight gavage, administration every day 1 time, successive administration 4d.Normal group and model group give the equal-volume distilled water.Before the last administration, water is can't help in the 16-18h fasting, 60min after the last administration, and except Normal group, all the other respectively organize equal lumbar injection 1% glacial acetic acid normal saline solution 0.1ml/10g, Normal group injection equal-volume normal saline.After giving acetic acid, observe immediately and record mice and the incubation period of writhing and the number of times of the interior mice generation writhing response of 15min occur.The animal that does not occur writhing in 15min is designated as 900s its incubation period.
30min after injection glacial acetic acid normal saline solution (Normal group injecting normal saline), all animal tail vein injection 0.5% azovan blue solution 0.2ml/10g take off cervical vertebra and put to death after 20min.The 2ml normal saline is injected the abdominal cavity, after soft abdominal part 30 times, collect peritoneal fluid, then divide the washing abdominal cavity 2 times with the 4ml normal saline, 2ml, collect respectively peritoneal fluid at every turn, is settled to 8ml after merging the peritoneal fluid of 3 times.Peritoneal fluid through the centrifugal 15min of 3000rpm, is got supernatant in 630nm place colorimetric, read absorbance.And take the Evans blue of variable concentrations as standard, the drawing standard curve calculates azovan blue content in peritoneal fluid.
Writhing suppression ratio (%)=(writhing number of times model group-writhing number of times medicine group)/writhing number of times model group * 100
Azovan blue seepage discharge suppression ratio (%)=(azovan blue seepage discharge model group-azovan blue seepage discharge medicine group)/azovan blue seepage discharge model group * 100
2.2 the impact of the rat toes swelling that the present composition is induced formula Freund's complete adjuvant not
The SD rat, male, 70.Left back pawl ankle every rat makes marks with stroke one horizontal line, measure sufficient volume with volumetric method, be divided at random by left back toes volume measurements: 1. Normal group, 2. model group, 3. aspirin 200mg/kg, 4. SHUANGHUANGLIAN KOUFUYE 6ml/kg, 5. present composition 1.75g crude drug/kg, 6. present composition 3.5g crude drug/kg, 7. present composition 7.0g crude drug/totally 7 groups of kg groups, 10 every group.Each group gives medicine 1ml/100g body weight by the body weight gavage, administration every day 1 time, successive administration 3d.Normal group and model group gavage equal-volume distilled water.Carried out toes swelling modeling on the 4th day and measure the swelling test, before test, water is can't help in the 16-18h fasting.Before giving the adjuvant modeling, toes basis volumetric values before the left back toes volumetric values of measuring every rat according to above-mentioned same method is as its modeling.Then, except Normal group, all the other are only respectively organized the equal subcutaneous injection Freund's complete adjuvant of the left back toes of rat 0.2ml/ so that are scorching, each administration group administration respectively simultaneously 1 time.The isometric normal saline of toes injection of normal control treated animal.1.0h, 2.0h, 4.0h, 6.0h measure respectively the left back toes volumetric values of rat after administration.Each group of 3.0h is strengthened administration 1 time again after modeling.Every animal cause toes solvent after inflammation all scorching with its causing separately before the toes volumetric ratio, calculate toes volume difference.
Toes volumetric values before the left back toes volumetric values of different time points after each time point toes volume difference=cause is scorching-cause is scorching
Toes volume difference suppression ratio (%)=(toes difference in volume value model group-toes volume difference medicine group)/toes difference in volume value model group * 100
3 experimental results
3.1 the impact of the present composition on the mice capillary permeability
The results are shown in Table 5.
The inhibitory action that the mouse writhing reaction that table 5 present composition Dichlorodiphenyl Acetate is induced and abdominal cavity capillary permeability increase
Annotate: compare * * P＜0.01, * * * P＜0.001 with model group; In () for discarding the number of animals after the haemolysis animal
Table 5 result shows, positive drug aspirin and SHUANGHUANGLIAN KOUFUYE all can obviously suppress inflammatory exudation (P＜0.001, P＜0.01), aspirin still has obvious analgesic activity, show as incubation period and less writhing number of times (P＜0.01, P＜0.001) that obvious prolongation writhing occurs.Present composition 2.5g crude drug/kg and 5.0g crude drug/kg dosage all can obviously suppress the stomachache writhing response that lumbar injection acetic acid causes, the writhing number of times obviously reduces (P＜0.01) than model group, the suppression ratio of 2 dosage groups is respectively 38.7%, 41.6%; And can significantly suppress the abdominal cavity capillary permeability increase that acetic acid causes, and reducing the seepage discharge (comparing P＜0.01 with model group) of azovan blue, suppression ratio is respectively 32.9%, 30.5%.
Result shows that the present composition has obvious analgesic and anti-inflammatory effects.
3.2 the impact of the rat toes swelling that the present composition is induced formula Freund's complete adjuvant not
The results are shown in Table 6.
The impact of the rat toes swelling that table 6 present composition is induced formula Freund's complete adjuvant not (
N=10)
Annotate: compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with model group
Table 6 result shows, the toes swelling of model group is very obvious, and after modeling, 1h is visible obvious tumefaction, extends in time, and the swelling degree increases, and continues at least 6h (each time point and Normal group more all have notable difference, P＜0.001) after administration; Positive drug aspirin and SHUANGHUANGLIAN KOUFUYE all after administration 2h begin to demonstrate the antiphlogistic effects of obvious inhibition swollen tissue.The present composition 1.75,3.5,3 dosage groups of 7.0g crude drug/kg all present the effect that alleviates toes swelling in various degree.Wherein, the onset time of 7.0g crude drug/kg dosage group is 2.0h after medicine, and 3.5g crude drug/kg dosage group onset time is 4.0h after medicine, and the onset time of 1.75g crude drug/kg dosage group is slower, is 6h after administration.3.5g the effect of crude drug/kg and 7.0g crude drug/kg dosage group is maintained until 6h after administration at least.After the present composition 1.75,3.5, the modeling of 3 dosage groups of 7.0g crude drug/kg, effect in 6 hours is the most obvious, suppression ratio is respectively 27.6,32.4,43.2% (relatively distinguishing P＜0.01, P＜0.001, P＜0.001 with model group), and demonstrates certain dose-effect relationship.
Result shows that the present composition has obvious antiinflammatory action.
The specific embodiment
Embodiment 1 granule
Raw material: Flos Lonicerae 600g, Flos Trollii 600g, Gentiana dahurica flower 200g, Radix Isatidis 1000g;
Preparation method: get crude drug, add 7 times of amount 70% alcohol reflux three times, each 1 hour; Merge extractive liquid, filters, and reclaims ethanol and is concentrated into the thick paste that relative density is 1.30～1.34 (50 ℃ of surveys); Get thick paste drying under reduced pressure below 80 ℃, be ground into fine powder, add adjuvant, mixing; Dry-pressing is granulated, 1000g processed altogether, and get final product;
Specification: every bag of 5g;
Usage and consumption: 3 times on the one, each 1 bag.
Embodiment 2 tablets
Raw material: Flos Lonicerae 550g, Flos Trollii 650g, Gentiana dahurica flower 180g, Radix Isatidis 1100g;
Preparation method: get crude drug, add 5 times of amount 90% alcohol reflux 2 times, each 1.5 hours; Merge extractive liquid, filters, and reclaims ethanol and is concentrated into the thick paste that relative density is 1.30～1.34 (50 ℃ of surveys); Get the thick paste drying under reduced pressure, be ground into fine powder, add conventional adjuvant, mixing; Dry-pressing is granulated, tabletting, and get final product.
Embodiment 3 capsules
Raw material: Flos Lonicerae 650g, Flos Trollii 550g, Gentiana dahurica flower 220g, Radix Isatidis 900g;
Preparation method: get crude drug, add 10 times of amount 50% alcohol reflux 4 times, each 1 hour; Merge extractive liquid, filters, and reclaims ethanol and is concentrated into the thick paste that relative density is 1.30～1.34 (50 ℃ of surveys); Get the thick paste drying under reduced pressure, be ground into fine powder, add conventional adjuvant, mixing; Dry-pressing is granulated, and incapsulates, and get final product.
Embodiment 4 powders
Raw material: Flos Lonicerae 500g, Flos Trollii 700g, Gentiana dahurica flower 150g, Radix Isatidis 1200g;
Preparation method: get crude drug, add 8 times of amount 60% alcohol reflux 3 times, each 1 hour; Merge extractive liquid, filters, and reclaims ethanol and is concentrated into the thick paste that relative density is 1.30～1.34 (50 ℃ of surveys); Get thick paste technique routinely, add conventional adjuvant to make powder.
Embodiment 5 oral liquids
Raw material: Flos Lonicerae 700g, Flos Trollii 500g, Gentiana dahurica flower 250g, Radix Isatidis 800g;
Preparation method: get crude drug, add 6 times of amount 75% alcohol reflux 2 times, each 1.5 hours; Merge extractive liquid, filters, and adds conventional adjuvant to make oral liquid.
Embodiment 6
Flos Lonicerae extract 500g, Flos Trollii extract 700g, Gentiana dahurica flower extract 150g, Radix Isatidis extract 1200g;
Get the pulverizing of said extracted thing and be fine powder, mixing is according to the agent of common process granulation;
Described Flos Lonicerae extract, Flos Trollii extract, Gentiana dahurica flower extract and Radix Isatidis extract are respectively Flos Lonicerae, Flos Trollii, Gentiana dahurica flower, Radix Isatidis with 7 times of amount 70% alcohol reflux three times, each 1 hour, reclaim the extract that ethanol obtains after merge extractive liquid.
Embodiment 7
Flos Lonicerae extract 600g, Flos Trollii extract 600g, Gentiana dahurica flower extract 200g, Radix Isatidis extract 1000g; Get the pulverizing of said extracted thing and be fine powder, mixing is made tablet according to common process;
Described Flos Lonicerae extract, Flos Trollii extract, Gentiana dahurica flower extract and Radix Isatidis extract are respectively Flos Lonicerae, Flos Trollii, Gentiana dahurica flower, Radix Isatidis with 7 times of amount 70% alcohol reflux three times, each 1 hour, reclaim the extract that ethanol obtains after merge extractive liquid.
Embodiment 8
Extracting honeysuckle 135g, Flos Trollii 135g, Gentiana dahurica flower 45g, Radix Isatidis 225g adds 10 times of decoctings and boils 3 times, and each 1 hour, filter, merging filtrate carries out precipitate with ethanol with 60% ethanol, reclaims ethanol and get final product.
Embodiment 9
Extracting honeysuckle 135g, Flos Trollii 135g, Gentiana dahurica flower 45g, Radix Isatidis 225g, with 60% alcohol reflux 3 times, solvent load is respectively 8,5,5 times of amounts, and each 1 hour, filter, merging filtrate reclaims ethanol and get final product.
Embodiment 10
Extracting honeysuckle 135g, Flos Trollii 135g, Gentiana dahurica flower 45g, Radix Isatidis 225g, wherein Flos Lonicerae, Flos Trollii, the merging of Gentiana dahurica flower are measured 60% alcohol reflux 3 times, each 1 hour with 6 times; 10 times of decoctings of Radix Isatidis 3 times, each 1 hour, merge extractive liquid, was with 60% ethanol precipitate with ethanol, and the ethanol part merges with three kinds of colored class extracting solution, reclaims ethanol and get final product.
Embodiment 11 determination of chlorogenic acid
The preparation of need testing solution: get respectively each 0.1g of sample of embodiment 9 and 10, precision weighing is put in the 25ml measuring bottle, adds methanol supersound process 20min, then adds methanol to scale, shakes up, and filters; Get subsequent filtrate 3.0ml, evaporate to dryness, residue add 50% methanol makes dissolving, is transferred in the 10ml volumetric flask, adds 50% methanol to scale and shakes up, and get final product.
Reference substance solution: get the chlorogenic acid reference substance and add the chlorogenic acid reference substance solution that methanol is made 0.04064mg/ml.
Chromatographic condition: chromatographic column: kromasil-C18 post; Mobile phase: acetonitrile-0.4% phosphoric acid solution (10: 90); Detect wavelength: 327nm; Flow velocity 1ml/min.
Experimental result sees Table 7.
Table 7 chlorogenic acid content
| Compositions | Chlorogenic acid content (mg/g Flos Lonicerae) | The chlorogenic acid rate of transform (%) |
| Embodiment 9 | 27.07 | 79.85 |
| Embodiment 10 | 26.75 | 78.91 |
The impact of embodiment 12 embodiment 8-10 compositions influenza virus infected life spans
1 test material
Test sample: the compositions (being 1g crude drug/ml extract) of embodiment 8-10 preparation ,-20 ℃ of Refrigerator stores, the used time is made into 0.6g crude drug/ml with distilled water.
Positive control drug: the oseltamivir phosphate capsule crude drug, be purchased from PVG Ying Xuan company, the used time is mixed with 1.25mg/ml with distilled water.
Strain: influenza virus is provided by Chinese department of Chinese medicine fundamental research institute of institute Viral Laboratory.
Laboratory animal: mice, strain: KM, male, animal origin: Beijing Vital River Experimental Animals Technology Co., Ltd., the animal quality certification number: SCXK (capital) 2007-0001.
2 test methods
2.1 dosage setting:
Test sample administration group is established respectively 1 dosage, i.e. 24g crude drug/kg; The dosage of positive drug oseltamivir phosphate capsule is 50mg/kg/d, for pressing the clinical equivalent dosage of body surface area conversion.
2.2 animal grouping
Selective body focuses on the healthy KM mice between 12-14g, is divided at random 6 groups: 1. normal control, and 2. model group, 3. oseltamivir phosphate capsule 50mg/kg/d, 4. embodiment is 8 groups, and 5. embodiment is 9 groups, and 6. embodiment is 10 groups, 12 of matched groups, 21 of other groups.
2.3 mouse infection is viral
Each is organized mice and slightly anaesthetizes with ether, with 15 times of LD
50Influenza virus drop nose infecting mouse, every collunarium volume is 60 μ l, every nostril 30 μ l.
2.4 route of administration
Each extract group of the model group of this test, positive drug group and the present invention all adopts the gastric infusion method.
2.5 medication and administration number of times, time:
The all beginning administrations in 1 day before the zoogenetic infection influenza virus of each extract group of the present invention, negative control and positive controls, every day, gastric infusion was 2 times, each administration of upper and lower noon 1 time, the administration volume is the 0.2ml/10g body weight, continuous 5 days.Negative control and Normal group gavage give with the volume distilled water.
2.6 observation index
The average survival time natural law of each group of record also calculates rate elongation.
3 statistical analysiss
The evaluation of life span adopts the average survival time natural law to assess, and data represent with means standard deviation, relatively use one factor analysis of variance (ANOVA) between group.
4 experimental results
The results are shown in Table 8.
The impact of table 8 present composition different process influenza virus infected survival day (
My god)
| Group | Dosage (g/kg) | Survival day (my god) | Rate elongation (%) |
| Matched group | - | 14.00±0.00 *** | - |
| Model group | - | 5.56±2.37 | - |
| The oseltamivir phosphate capsule group | 0.05 | 9.67±3.46 | 73.92 |
| Embodiment 8 | 24 | 6.70±3.34 | 20.50 |
| Embodiment 9 | 24 | 7.87±3.98 | 41.55 |
| Embodiment 10 | 24 | 6.63±4.06 | 19.24 |
Annotate: rate elongation (%)=(survival day
The medicine group-survival day
Model group)/survival day
Model group* 100 by as seen from Table 8, and embodiment 8-10 medicine group all can extend the life span of the viral mice of infection.
Claims (10)
1. Chinese medicine composition for the treatment of anemopyretic cold is characterized in that the crude drug of this Chinese medicine composition consists of:
Flos Lonicerae 400-800 weight portion Flos Trollii 400-800 weight portion
Gentiana dahurica flower 100-300 weight portion Radix Isatidis 700-1300 weight portion.
2. Chinese medicine composition as claimed in claim 1 is characterized in that the crude drug of this Chinese medicine composition consists of:
Flos Lonicerae 550-650 weight portion Flos Trollii 550-650 weight portion
Gentiana dahurica flower 180-220 weight portion Radix Isatidis 900-1100 weight portion.
3. Chinese medicine composition as claimed in claim 1 is characterized in that the crude drug of this Chinese medicine composition consists of:
Flos Lonicerae 600 weight portion Flos Trollii 600 weight portions
Gentiana dahurica is spent 200 weight portion Radix Isatidis 1000 weight portions.
4. preparation method as Chinese medicine composition as described in claim 1-3 any one, the method is: get in proportion crude drug, adopt organic solvent extraction.
5. preparation method as claimed in claim 4, is characterized in that, described organic solvent is the ethanol of 50-90%.
6. as the preparation method of Chinese medicine composition as described in claim 1-3 any one, the method is: get in proportion crude drug, adopt the decoction and alcohol sedimentation technique preparation; Wherein said alcohol precipitation concentration is 40-90%.
7. as the preparation method of Chinese medicine composition as described in claim 1-3 any one, the method is: get in proportion crude drug, wherein three kinds of colored class medical materials adopt the organic solvent united extraction, get extract A; Radix Isatidis adopts separately decoction and alcohol sedimentation technique to be prepared into extract B; United extraction thing A and B, and get final product; Wherein said organic solvent is the ethanol of 50-90%; Described alcohol precipitation concentration is 40-90%.
8. as the application of Chinese medicine composition as described in claim 1-3 any one in preparation treatment anemopyretic cold medicine.
9. suppress application in influenza virus property medicine as Chinese medicine composition as described in claim 1-3 any one in preparation.
10. as the application of Chinese medicine composition as described in claim 1-3 any one in the preparation anti-inflammatory drug.
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