Summary of the invention
By extracting with rational process, preparing above four Chinese medicine thing, keep to greatest extent and the purification active constituents of medicine, thereby the stronger medicine of exploitation curative effect is one of purport of the present invention.
For this reason, one of the object of the invention provides a kind of extract with antiinflammation and preparation method thereof.Another purpose of the present invention provides a kind of medicine with antiinflammation, especially has the oral liquid and the preparation thereof of antiinflammation.The 3rd purpose of the present invention is the clinical application that said medicine is provided.
Therefore, the technical solution used in the present invention is as follows:
A kind of extract with antiinflammation, calculate by weight, 1 part of Herba Corydalis Bungeanae, 1.5 parts of Radix Isatidis, 4 parts of Herba Taraxacis, 0.05~0.15 part of 1.5 parts of Radix Scutellariaes or baicalin, it is characterized in that described extract is prepared from by the method that comprises following steps:<1〉Herba Corydalis Bungeanae, the water extract that Radix Isatidis and Herba Taraxaci obtain with water extraction, add ethanol and make that to contain alcohol amount be 60%~85%, remove insoluble matter, solution removes to desolvate and obtains extract,<2〉to step<1〉add the baicalin that described baicalin or described Radix Scutellariae obtain through extraction in the resulting extract.
Extract of the present invention can directly add the baicalin of ormal weight on forming, also can obtain baicalin by method extraction separation common in the prior art with the Radix Scutellariae of ormal weight.The extraction ratio of baicalin in the radix scutellariae medicinal materials, relevant with content of baicalin in the radix scutellariae medicinal materials and extraction separation method, yield is usually in 3%~10% scope.As one of preferred version, Radix Scutellariae of the present invention extracts with the method that comprises following steps and obtains baicalin:<1〉Radix Scutellariae obtains water extract with water extraction;<2〉step<1〉resulting water extract adds ethanol makes the alcohol amount of containing 40%~60%, removes insoluble matter, and solution reclaims ethanol;<3〉step<2〉reclaim solution adjust pH behind the ethanol to acid, collect precipitate, obtain baicalin.
Wherein, preferred Radix Scutellariae drops in the boiling water and extracts.
More preferably Radix Scutellariae is dropped in the boiling water and decoct, decocted 5~30 minutes earlier at every turn, to about 6.5, decocted again 0.5~1 hour with the lye pH adjustment value.
As one of preferred version, wherein above-mentioned described step<3〉in, after the solution after the precipitate with ethanol step reclaimed alcohol solvent, solution was in 60~90 ℃ of insulations, with mineral acid or organic acid adjust pH 1~2; Be preferable over 80 ℃ of insulations, with hydrochloric acid adjust pH 1.5.
The present invention also provides a kind of medicine with antiinflammation, and it comprises above-mentioned described extract as active component and acceptable accessories.Wherein, as one of specific embodiment, the invention provides a kind of oral liquid with antiinflammation, it comprises above-mentioned described extract as active component, also can contain sweeting agent (for example steviosin) as correctives.
As preferred version, described oral liquid, wherein, prescription consists of: Herba Taraxaci 500 grams, Herba Corydalis Bungeanae 125 grams, Radix Scutellariae 188 grams, Radix Isatidis 188 grams are made 1000 milliliters oral liquid.
More preferably above-mentioned oral liquid, prepare by the method that comprises following steps:<1 Herba Taraxaci, Radix Isatidis and Herba Corydalis Bungeanae water boiling and extraction, extracting solution is concentrated into the clear paste of 1.13~1.15 (60~70 ℃ of heat are surveyed), add ethanol and make the alcohol amount about 75% that contains, place after-filtration, filtrate recycling ethanol adds water to 500 milliliters, place after-filtration, filtrate is standby with 10% sodium hydroxide adjust pH about 6.5;<2〉Radix Scutellariae drops into to decoct in the boiling water and extracts, the solution pH value about 6.5 in the decoction process, extracting solution is concentrated into relative density 1.08~1.11 (60~70 ℃ heat survey), and adjust pH is about 6.5, adds ethanol and makes the alcohol amount about 50% that contains, place after-filtration, behind the filtrate recycling ethanol, in about 70~80 ℃ of insulations, with hydrochloric acid adjust pH about 1.5, place, collect the precipitate of baicalin;<3〉with step<2〉resulting baicalin adds 500 ml waters in 70~80 ℃ of insulation dissolvings, and is standby with 10% sodium hydroxide adjust pH about 6.5;<4〉with above-mentioned steps<1〉and<3〉resulting solution merging, add 10 gram steviosin, add water to 1000 milliliters.
Above-mentioned described extract or the medicine with antiinflammation of the present invention can be used for preparing the medicine with heat-clearing and toxic substances removing or antiinflammatory or detumescence effect.Especially, can be used for preparing the medicine that is used for the treatment of diseases such as parotitis, pharyngitis, tonsillitis or furuncle and phyma.
Extract of the present invention, adopt Herba Taraxaci, Herba Corydalis Bungeanae and Radix Isatidis water extract-alcohol precipitation in the preparation method and add the technology of baicalin, product is at aspects such as color and luster, active constituent content and curative effect of medication, has unexpected remarkable result, the oral liquid formulations that is prepared from, the clarity of product and stability also have excellent effect.
The specific embodiment
Following specific embodiment is to set forth content of the present invention for further, and should be understood that claim protection domain of the present invention is construed as limiting effect.
Embodiment 1
1A) Herba Taraxaci 40 grams, Radix Isatidis 15 grams, Radix Scutellariae 15 grams, Herba Corydalis Bungeanae 10 grams.Wherein, Herba Taraxaci, Radix Isatidis, Herba Corydalis Bungeanae samadhi medical material decoct with water secondary, and each 1 hour, extracting solution was concentrated in right amount, add ethanol and make that to contain alcohol amount be 70%, place the back and filter, and filtrate recycling ethanol concentrates, and obtains the water extracting alcohol hypostasis; Other gets Radix Scutellariae, and drop in the boiling water and decoct secondary, each 1 hour, merge extractive liquid,, be concentrated in right amount, add ethanol and make and contain the alcohol amount and reach 50%, place after-filtration, behind the filtrate recycling ethanol, solution is 1.5 80 ℃ of insulations with the hydrochloric acid adjust pH, places after-filtration, makes baicalin.Baicalin is joined in the water extracting alcohol hypostasis that makes previously, obtain extract of the present invention (claiming " water alcohol+yellow method " extract).
1B) Herba Taraxaci 40 grams, Radix Isatidis 15 grams, Radix Scutellariae 15 grams, Herba Corydalis Bungeanae 10 grams.The four Chinese medicine material decocts with water secondary, and each 1 hour, extracting solution was concentrated in right amount, adds ethanol and makes that to contain alcohol amount be 70%, places the back and filters, and filtrate recycling ethanol concentrates, and obtains water extracting alcohol hypostasis (claiming " water alcohol method " extract).
1C) Herba Taraxaci 40 grams, Radix Isatidis 15 grams, Radix Scutellariae 15 grams, Herba Corydalis Bungeanae 10 grams.Wherein, Radix Scutellariae is according to 1A) method extracts and to obtain baicalin.Other three flavor medical materials are with 70% alcohol reflux twice, and each 1 hour, extracting solution reclaimed ethanol, are concentrated in right amount, add baicalin, obtain extract (title " backflow+yellow method " extract).
With above-mentioned 1A)~1C) the prepared extract of method, it is an amount of to add water, makes oral liquid, investigates stability of formulation.Carry out assay simultaneously respectively, measure content of baicalin.
Content assaying method: high performance liquid chromatography is measured.Chromatographic condition is: with octadecylsilane chemically bonded silica is filler; Methanol-water-phosphoric acid (44: 56: 0.2) is a mobile phase, and the detection wavelength is 280nm.Number of theoretical plate calculates by the baicalin peak should be not less than 2500.
The preparation of reference substance solution: it is an amount of that precision takes by weighing the baicalin reference substance, adds dissolve with methanol and make per 1 milliliter of solution that contains 50 micrograms, shakes up promptly.
The preparation of need testing solution: precision is measured 1 milliliter of prepared oral liquid, places 50 milliliters of measuring bottles, adds methanol and is diluted to scale, shakes up, promptly.
Accurate respectively reference substance solution and each 10 microlitre of need testing solution drawn inject chromatograph of liquid, measure promptly.
Stability and assay result are as shown in table 1 below.
Table 1 1A)~1C) the prepared extract measurement result of method
Sample | Clarity | Baicalin (mg/ml) | CV% |
1A) " water alcohol+yellow method " 1B) " water alcohol method " 1C) " backflow+yellow method " | Have after two weeks that a little precipitation is very fast promptly to have precipitation precipitation to be arranged after three days | 4.6 1.4 4.2 | 1.2 2.4 2.2 |
By above-mentioned test as can be seen, from the clarity and the content of baicalin aspect of product, the person is good all to add the baicalin with other; Wherein, water alcohol adds baicalin and adds baicalin for good than backflow again, and preparation is more convenient again.Therefore, the preparation of extract of the present invention, Herba Taraxaci, Radix Isatidis and Herba Corydalis Bungeanae adopt the water extraction Ethanol Treatment to obtain extract, add baicalin more in addition, and the extract active constituent content of gained significantly improves, and product stability is better.
Embodiment 2: the preparation oral liquid
Get Herba Taraxaci 500 grams, Radix Isatidis 188 grams, Herba Corydalis Bungeanae 125 grams decoct with water secondary, each 1 hour.Merge extractive liquid,, being concentrated into relative density is the clear paste of 1.13~1.15 (60~70 ℃), adds ethanol and makes that to contain alcohol amount be 75%, places 48 hours, filter, filtrate recycling ethanol adds water to 500 milliliters, places 48 hours, filter, filtrate is 6.5 with 10% sodium hydroxide solution adjust pH, and is standby.
Other gets Radix Scutellariae 188 grams, drops in the boiling water and decocts secondary, each decoction earlier 10 minutes, regulating pH value with 10% sodium hydroxide solution is 6.5, decocts merge extractive liquid, again 1 hour, be concentrated into relative density 1.08~1.11 (60~70 ℃), adjust pH to 6.5 adds ethanol and makes and contain alcohol amount and reach 50%, places 24 hours, filter, behind the filtrate recycling ethanol, add water and make into 1: 1, can remove by filter precipitation if any precipitation, solution is 80 ℃ of insulations, with the hydrochloric acid adjust pH is 1.5, is incubated half an hour, places after 24 hours, filter, precipitate is washed till neutrality with 70% ethanol, obtains the baicalin crude product, adds 500 milliliters in water, 80 ℃ of insulations, 10% sodium hydroxide adjust pH to 6.5 is used in dissolving simultaneously, and is standby.
Above-mentioned two kinds of solution are merged, add the steviosin of 10 grams, add water to 1000 milliliters, packing, circulation steam sterilization 30 minutes obtains oral liquid.
Prepared oral liquid is investigated through stability test, has good stability, and product is still clarified after three months.
Embodiment 3: anti-white mice acute inflammation pharmacological experiment study
1, experiment material
Animal: 18~22 gram Kunming kind white mice, male and female half and half.
Medicine: embodiment 2 prepared oral liquids.
2, method
36 of healthy mices are divided into 3 groups at random: the oral liquid group of the present invention (42g/kg body weight) of blank group, heavy dose and low dose of oral liquid group of the present invention (14g/kg body weight).Every group 12, male and female half and half.
The administration 1 time respectively of cause acute inflammation preceding 6 hours and 1 hour, each blank group and administration group white mice are irritated stomach 0.21ml/10g body weight normal saline and corresponding experiment medicinal liquid respectively.The each dosage of heavy dose of oral liquid group of the present invention is the 21g/kg body weight, and the each dosage of low dose of oral liquid group of the present invention is the 7g/kg body weight.
The experiment white mice respectively at administration after 1 hour, splash into 0.07ml dimethylbenzene at the auris dextra exterior feature, put to death in 30 minutes, cut left and right auricle, be that 8 millimeters circular card punch is respectively laid a circular auricle in left and right sides auricle same position with diameter, weigh.The difference that deducts left auricle weight with auris dextra sheet weight is as inflammation swelling degree index, and is benchmark with blank group inflammation swelling degree, calculates each administration group inflammation suppression ratio.
3, result
Experimental result sees Table 2.As shown in Table 2, white mice ear acute inflammation due to the xylol, 14g/kg and 42g/kg oral liquid group of the present invention all has the obvious anti-inflammatory and anti effect, and suppression ratio reaches 20.6% and 38.8% respectively, compares with the blank group, has significant difference.
Table 2 oral liquid xylol of the present invention causes the acutely inflamed influence of white mice ear
Group | Dosage (g/kg body weight) | Inflammation swelling degree (x ± SD) | Suppression ratio (%) |
The heavy dose of administration group of blank group | 42 | 16.0±3.9 9.8±3.8** | 38.8 |
Low dose of administration group | 14 | 12.7±4.0* | 20.6 |
Annotate: compare with the blank group, the t check,
*P<0.05,
*P<0.01, n=12
Embodiment 4: the experiment of the subcutaneous mixed infection of anti-white mice
1, material
Medicine: embodiment 2 prepared oral liquids.Homemade commercially available Pianomide is as positive control drug.Preparation process is as follows:
Radix Scutellariae 450 grams, Herba Taraxaci 1200 grams, Herba Corydalis Bungeanae 300 grams, Radix Isatidis 450 grams.More than four flavors, Radix Scutellariae 250 gram is ground into fine powder, sieves, and is standby.All the other Radix Scutellariaes 60% ethanol extraction reclaims ethanol, is condensed into cream.Herba Taraxaci, Herba Corydalis Bungeanae decoct with water secondary, and collecting decoction filters.Radix Isatidis decocts with water back warm macerating secondary, filters, and merging filtrate adds above-mentioned decocting liquid, is condensed into thick paste, adds above-mentioned ethanol condensed cream, Baical Skullcap Root and right amount of auxiliary materials, and mixing is made granule, and drying is pressed into 2660.
For trying antibacterial: escherichia coli, bacteroides fragilis
White mice: body weight 18~20 grams, male and female half and half, random packet.
2, method
Escherichia coli are diluted to every milliliter with 5% gastric Mucin contain 1.5 * 10
8Individual bacterium suspension; Bacteroides fragilis is diluted to every milliliter with saline and contains 2 * 10
8Individual bacterium suspension.Before the experiment that above-mentioned two kinds of solution equal-volumes mixing back is standby.
Get 140 of white mice, be divided into 7 groups, 20 every group, in infecting preceding 24 hours, clothes are for reagent or contrast 0.5 milliliter of medicine, inject 0.2 milliliter by per 20 gram body weight in carrying on the back under the hypodermis with the above-mentioned examination bacterium mixed liquor that supplies on 1st, and rechallenge once after 1 hour.The blank group, then with the physiologic saline for substitute medicine, successive administration 9 days.Observe the subcutaneous infection site of white mice that swelling stroke and size thereof are arranged not and compared.
The results are shown in Table 3 and 4.
The subcutaneous infection effect of the anti-white mice of table 3 relatively
Group | Dosage (mg//day) | White mice number (individual) | Swelling forms number (individual) | Swelling rate (%) | Compare p with the normal saline group |
Oral liquid group Pianomide matched group of the present invention | 500 250 125 500 250 125 | 20 20 20 20 20 20 | 10 12 20 12 18 20 | 50 60 100 60 90 100 | <0.01 <0.01 >0.05 <0.01 >0.05 >0.05 |
The normal saline group | | 20 | 20 | 100 | |
As can be seen from Table 3: oral liquid group of the present invention is compared with the Pianomide positive controls, and all by tangible anti-infectious function, but of the present invention group of curative effect obviously is better than the Pianomide positive controls, and marked difference (p<0.05) is arranged.
The subcutaneous influence that infects back formation abscess area of table 4 pair white mice
Group | Dosage (mg//day) | White mice number (individual) | Swelling forms number (individual) | Swelling area X ± S (mm
2)
| Compare p with the normal saline group |
Oral liquid group Pianomide matched group of the present invention | 500 250 125 500 250 125 | 20 20 20 20 20 20 | 10 12 20 12 18 20 | 19.10±2.70 30.58±2.16 81.00±2.86 23.75±3.31 80.00±3.49 83.10±4.70 | <0.01 <0.01 >0.05 <0.01 >0.05 >0.05 |
The normal saline group | | 20 | 20 | 88.20±1.90 | |
As can be seen from Table 4: oral liquid group of the present invention is compared with the Pianomide positive controls, and all by tangible influence, but of the present invention group of curative effect obviously is better than the Pianomide positive controls, and marked difference (p<0.01) is arranged to the formation of white mice abscess.
The present invention is described according to preferred embodiment.Should be understood that the description of front and embodiment are just to illustrating the present invention.Under prerequisite without departing from the spirit and scope of the present invention, those skilled in the art can design multiple alternative of the present invention and improvement project, and it all should be understood to be within protection scope of the present invention.