CN103446220B - Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to composition of medicine, preparation method and application - Google Patents
Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to composition of medicine, preparation method and application Download PDFInfo
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Abstract
The invention provides a kind of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to composition of medicine, described composition of medicine is mixture or its extract composition of Herba Artemisiae Annuae and Radix Tripterygii Wilfordii two kinds of medical materials; Body of the present invention provides the extract of a kind of medicine to Herba Artemisiae Annuae Radix Tripterygii Wilfordii composition of medicine.This extract is relatively and Radix Tripterygii Wilfordii extract, lower to the toxicity of Liver and kidney, simultaneously relatively with commercially available Radix Tripterygii Wilfordii pharmaceutical preparation treat in rheumatoid arthritis disease disease better active.
Description
Technical field
The present invention relates to a kind of Chinese medicines to combination, relate to a kind of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine in particular to composition of medicine, preparation method and application.
Technical background
Radix Tripterygii Wilfordii (Tripterygiumwilfordii) is warm in nature, bitter and puckery flavor.There is the effects such as expelling wind and removing dampness, relaxing muscles and tendons and activating QI and blood in the collateral, reducing swelling and alleviating pain, parasite killing removing toxic substances.Modern study finds to have antiinflammatory, immunosuppressant, antifertility antitumor, antibacterial, pain relieving isoreactivity.The aspect such as rejection, autoimmune disease, nephrotic syndrome, cancer of discovered in recent years Radix Tripterygii Wilfordii to organ transplantation is evident in efficacy, clinically for the treatment of the difficult diseases such as rheumatic arthritis, rheumatoid arthritis, traumatic injury, glomerulonephritis, lupus erythematosus, the nephrotic syndrome.Up to now, Chinese scholars has been separated and has obtained about 7O kind composition, mainly alkaloids, Diterpenes (as triptolide, i.e. Radix Tripterygii Wilfordii lactone alcohol), triterpenes, sesquiterpenoids and polysaccharide from tripterygium plant.Since Radix Tripterygii Wilfordii is nearly over half a century there is one of maximum Chinese herbal medicine of poisoning in report.Along with people are to the increase of deepen continuously exploration and the clinical practice of its effective ingredient and relevant pharmacological action, its toxic and side effects and influence factor thereof also receive publicity more.Herein by recent years about the research of Radix Tripterygii Wilfordii toxicity is summarized.
Mainly based on cardiac muscle, nervous system, kidney damage, even there is conscience hemorrhage or downright bad in Tripterygium wilfordii Poisoning, rapidly, mortality rate is high in development.Symptom is that Progressive symmetric erythrokeratodermia increases the weight of, first meeting Nausea and vomiting, stomachache, diarrhoea etc., then occur that thready and rapid pulse and weak, blood pressure reduce, dizzy, headache, weak, nervous, irritated so that tic, then may there is lumbago, oliguria, hematuria, albuminuria, non-protein nitrogen raise, finally lethal because of the lists such as the damaged nerve cell of acute renal failure, circulatory failure, central nervous system and serious bone marrow depression or multiple organs failure.
Tripterygium Preparations has become one of common drug of clinical treatment RA due to determined curative effect.But the therapeutic dose of Radix Tripterygii Wilfordii and toxic dose are closely, long-term prescription toxicity is easily accumulated, and causes the irreversible pathological changes of internal organs such as genitals, liver, kidney.And its drug effect, toxicity and dosage have obvious dose-effect relationship, by reducing Radix Tripterygii Wilfordii dosage be the effective thinking reaching attenuation synergistic according to certain compatibility the form of the rules compound recipe.Animal and clinical experiment display, arteannuin and derivant thereof have treatment RA effect preferably, and its toxicity is minimum.This research, by the tripterygium glycosides of low dose and artesunate compatibility, has great importance for exploration safety, rheumatoid arthritis medicine that is efficient, low toxicity.
Summary of the invention
For solving the huge toxicity problem that Tripterygium Preparations exists, the invention provides a kind of Herba Artemisiae Annuae (Herba Artemisiae Annuae of the present invention is the dry aerial parts of Herba Artemisiae annuae ArtemisiaannuaL.) and Radix Tripterygii Wilfordii medicine to composition of medicine, described composition of medicine is mixture or its extract composition of Herba Artemisiae Annuae and Radix Tripterygii Wilfordii two kinds of medical materials.
Described composition of medicine is that Herba Artemisiae Annuae and Radix Tripterygii Wilfordii two kinds of medical materials mix, and then adopts process for super-critical extracting to prepare extract.
The weight ratio of described Herba Artemisiae Annuae and tripterygium wilfordii is 1:1-3.
The weight ratio of described Herba Artemisiae Annuae and tripterygium wilfordii is 1:1.
Described method is adopt supercritical extraction to extract after being mixed with tripterygium wilfordii by Herba Artemisiae Annuae.
Described Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to the preparation method of composition of medicine, and the condition of described supercritical extraction is: CO
2flow is 4.6Lh
-1, extraction temperature 45 DEG C, extracting pressure 25MPa, employing ethanol is entrainer, and entrainer consumption is 2mLg
-1, extraction time 100min.
Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine described above is to the application of composition of medicine in preparation treatment rheumatoid arthritis agents.
Advantageous Effects of the present invention is: body of the present invention provides the extract of a kind of medicine to Herba Artemisiae Annuae Radix Tripterygii Wilfordii composition of medicine.This carries extract relatively and Radix Tripterygii Wilfordii extract, lower to the toxicity of Liver and kidney, simultaneously relatively with commercially available Radix Tripterygii Wilfordii pharmaceutical preparation treat in rheumatoid arthritis disease disease better active.
Detailed description of the invention
Embodiment 1 Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is investigated process for super-critical extracting
Take Herba Artemisiae Annuae and 5 parts, tripterygium wilfordii (1:1) mixture of 100.0g drying, use methanol, ethanol, 75% ethanol, ethylene glycol and ethyl acetate as entrainer respectively, extract, get portion without entrainer in contrast;
Dose in supercritical extraction reactor by above-mentioned Herba Artemisiae Annuae and tripterygium wilfordii mixture, follow procedure heats up and pressurizes, and carries out dynamic extraction, CO
2flow is 4.6Lh
-1, extraction temperature 45 DEG C, extracting pressure 25MPa, entrainer consumption is 2mLg
-1, quantitatively pump into entrainer, extraction time 100min; Analytically product collected by still, reclaim under reduced pressure organic solvent, and obtains Herba Artemisiae Annuae tripterygium wilfordii supercritical extract, and concrete outcome is as shown in table 1:
The impact that the different entrainer of table 1 extracts Radix Tripterygii Wilfordii lactone alcohol and artesunate
As can be seen from Table 1, methanol, ethanol, 75% ethanol, ethylene glycol and ethyl acetate five kinds of entrainers improve solute all to a certain extent at SC-CO
2in dissolubility, but four kinds of different entrainers are different to the increase rate of solid solute dissolubility, and ethanol is the strongest and 75% ethanol is the strongest, methanol takes second place, third, acetic acid ethyl dissolution amplitude is the most weak for ethylene glycol, and this and the association formed between entrainer and solute are closely related.Methanol, ethanol, ethylene glycol and ethyl acetate are all polar substancess, can form hydrogen bond, with solute generation hydrogen bond association, form associated complex, thus improve Radix Tripterygii Wilfordii lactone alcohol (triptolide) and artesunate isoreactivity composition dissolubility in a solvent; But because the polarity of five kinds of materials is different, the hydrogen bond of formation varies in size, thus to Radix Tripterygii Wilfordii lactone alcohol and artesunate at supercritical CO
2the increase rate of middle dissolubility is also different.
The introducing of entrainer gives SC-CO
2the application that abstraction technique is more wide, also brings two negative effects simultaneously.Here it is due to the use of entrainer, adds the difficulty of Separation and Recovery entrainer from extract.And owing to employing entrainer, make in some extracts, there be the residual of entrainer.This just loses SC-CO
2extraction does not have the advantage of dissolvent residual; Therefore, adopt ethanol as entrainer, the yield of the sterling of product is the highest, and residual harm is minimum.
The different extracting method of embodiment 2 is on the impact of the active component rate of transform
1, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to the preparation of ethanol extract
By Herba Artemisiae Annuae and tripterygium wilfordii (1:1) mixture 95% ethanol extraction 2 times, amount of alcohol is 8 times of medical material amount. extraction time is respectively 2,1.5h, 180 DEG C of boilings, and 130 DEG C keep micro-boiling, and is respectively 2,1.5h.After having extracted, medicinal residues are poured out, dry.Extracted twice thing merges.Concentrating under reduced pressure 95% ethanol extraction, temperature is 50 DEG C, and rotating speed is 34r/min, and pressure is-0.095MPa, is concentrated into 2L.Take out 500mL to keep sample, evaporate in water-bath as, temperature is 82.5 DEG C, then puts into baking oven and be dried into dry thing (temperature of baking oven is 42.5 DEG C), obtains Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to alcohol extract.
2, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to the preparation of supercritical extract
The method described in embodiment 1 of employing, takes medical material in the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:1), pulverizes, and adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract I.
The method described in embodiment 1 of employing, takes medical material in the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (3:1), pulverizes, and adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract II.
The method described in embodiment 1 of employing, takes medical material in the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:3), pulverizes, and adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract III.
The method described in embodiment 1 of employing, takes medical material in the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (4:1), pulverizes, and adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract IV.
The method described in embodiment 1 of employing, takes medical material in the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:4), pulverizes, and adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract V.
3, the preparation of tripterygium wilfordii supercritical extract
The method described in embodiment 1 of employing, pulverizes tripterygium wilfordii, adopts the ethanol of 75% as entrainer, prepares Radix Tripterygii Wilfordii supercritical extract.
3, the preparation of Artemisia annua supercritical extract
The method described in embodiment 1 of employing, by Herba Artemisiae annuae pulverizing medicinal materials, adopts the ethanol of 75% as entrainer, prepares Herba Artemisiae Annuae supercritical extract.
The lesions of liver and kidney effect of embodiment 4 pairs of mices is investigated
ICR mice, cleaning grade, male and female half and half, weight 18-22g, totally 180, mice is divided into 9 groups at random, is respectively: blank group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to alcohol extract group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract I; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract II group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract III group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract IV group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract V group, Radix Tripterygii Wilfordii supercritical extract group and Herba Artemisiae Annuae supercritical extract group.Often organize laboratory animal in two batches, often criticize 10, male and female half and half, a collection of administration 5d, a collection of administration 10d.Dosage is clinical equivalent amount (18.00g (crude drug)/kg), and blank group gives equal-volume normal saline, every day gastric infusion 1 time.
Stablize 3d before above-mentioned mouse experiment, the general status such as the ingesting of observation mice, feces, activity.Respectively at after experiment grouping the 5th day and administration 12h on the 10th (water 12h is can't help in fasting), blood is got to two batches of animal eye sockets lethal, in the centrifugal 10min of 3000r/min, get serum, measure ALT, AST and BUN, Scr level.
Application SPSSI6.0 statistical software.Measurement data with
represent.P<0.05 is that difference has statistical significance.
Table 2 respectively group liver function index compares
Compare with blank group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Table 2 result shows, after administration 5d, compare with blank group, to alcohol extract group, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, to supercritical extract III group and Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, ALT and the AST value to supercritical extract V group and Radix Tripterygii Wilfordii supercritical extract group all raises Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, and there were significant differences (P<0.05 or O.01).After administration 10d, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine all raises supercritical extract V group and Radix Tripterygii Wilfordii supercritical extract group ALT and AST value supercritical extract III group and Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine alcohol extract group, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, and and blank group compare that there were significant differences (P<0.05 or 0.01).Analyze above data to find, when the ratio of Herba Artemisiae Annuae and Radix Tripterygii Wilfordii is lower than 1:3, the ALT of 5d and 10d raises rapidly, and when 10d, AST also raises rapidly.
Table 3 respectively group renal function index compares
Compare with blank group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Analytical table 2 data are known, after administration 5d, to alcohol extract group, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, to supercritical extract III group and Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, the BUN value to supercritical extract V group and Radix Tripterygii Wilfordii supercritical extract group raises Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, and blank group compares, and there were significant differences (P<0.05 or 0.01).But the Ser impact of administration 5d on mice is little.After administration 10d, Artemisia Radix Tripterygii Wilfordii medicine to alcohol extract group, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract III group and Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract V group and Radix Tripterygii Wilfordii supercritical extract group; And and blank group compare that there were significant differences.Ethanol extract group Scr value raises, and blank group comparing difference significantly (P<0.05).
Embodiment 4
Female Wistar rats, 130 ~ 170g.Test preposition animal in laboratory endoadaptation environment 3 days, room temperature 18-2O DEG C, relative humidity 65%.At random animal is divided into 9 groups: 1. normal group, normally feeds; 2. model group, makes the rat arthritis model of II Collagen Type VI induction.3. commercially available Tripterygium Hypoglaucum Hutch Tablet group (commercially available group), rat arthritis model gives commercially available Tripterygium Hypoglaucum Hutch Tablet (the accurate word Z20027411 of traditional Chinese medicines) 20mg/d after making.4. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to alcohol extract group (alcohol extraction group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to alcohol extract after making, dosage 42g(crude drug)/kg.5. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract I group (supercritical I group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to supercritical extract I after making, dosage is 28g(crude drug)/kg.6. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract II group (supercritical II group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to supercritical extract II after making, dosage is 28g(crude drug)/kg.7. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract III group (supercritical III group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to supercritical extract III after making, dosage is 28g(crude drug)/kg.8. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract IV group (supercritical IV group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to supercritical extract IV after making, dosage is 28g(crude drug)/kg.9. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to supercritical extract V group (supercritical V group), and rat arthritis model gives Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine that embodiment 2 prepares to supercritical extract V after making, dosage is 28g(crude drug)/kg.
The rat arthritis model of II Collagen Type VI induction: acid-soluble II Collagen Type VI 50mL is dissolved in 0.1mol, in 25mL acetum, then 4 spend the night drips equal-volume complete Freund's adjuvant (CTA) grinding and evenly makes Emulsion and make collagen content be 1mg/mL under ice bath, inject 0.1 respectively at rat root of the tail portion, back and cervical region Intradermal, 0.2,0.2mL collagen breast (0.5mL/ only), use ethanol partly sterilised before injection.14th day with the 100 μ L/ CTA immune rat again containing collagen 1mg/mL only.Occur with rat hindleg red and swollen for the onset time, randomly draw rat investigation indices for each group and comprise the performance of inspection sign, local joint histopathology inspection, immunologic test, hematological examination, image analysis.
Method is affected on inflammation integration: experimental rat, after the immunity of secondary calf II Collagen Type VI, observes toes swelling situation, respectively by relevant requirements grade scale, keep the score, record once every other day, until experiment terminates the previous day, adopt every rat integration addition method.
The result result that affects of inflammation integration shows: 3rd, 5,7,9,11,13,15,18,21 days inflammation integrations all show, middle dosage group, small dose group compare with model group significant difference (P<0.05), heavy dose of group compares with model group significant difference (P<0.O1), illustrates that GSF has the inflammatory effect of the autoimmune arthritis significantly suppressing II Collagen Type VI to bring out.Specifically as shown in table 4.
Table 4 inflammation integration record
Note: compare with model group,
*p ﹤ 0.05,
*p ﹤ 0.01.
Method is affected on II Collagen Type VI induced arthritis SOD in serum content: undertaken by test kit description.
SOD content model group in result serum is affected on II Collagen Type VI induced arthritis SOD in serum content and compares there was no significant difference with Normal group, but alcohol extraction small dose group SOD in serum content is significantly higher than model group.Specifically as shown in table 5.
Table 5 affects II Collagen Type VI induced arthritis SOD in serum content
In sum, each group Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine has the pharmacological action of antiinflammatory, immunity moderation to supercritical extract; Clear and definite therapeutical effect is had to rheumatoid arthritis.Wherein Herba Artemisiae Annuae and tripterygium wilfordii (1:1), Herba Artemisiae Annuae and the activity of tripterygium wilfordii (1:3) two groups of medicines to compositions best, Herba Artemisiae Annuae and tripterygium wilfordii (1:4) take second place, Herba Artemisiae Annuae and tripterygium wilfordii (3:1) Herba Artemisiae Annuae and the activity of tripterygium wilfordii (4:1) two groups of medicines to compositions the poorest.Can know from above-mentioned, the ratio of Herba Artemisiae Annuae and tripterygium wilfordii is 1:1-3 is optimal proportion.
Claims (2)
1. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to a composition of medicine, and described composition of medicine is the extract composition of Herba Artemisiae Annuae and Radix Tripterygii Wilfordii two kinds of medical materials; It is characterized in that: described composition of medicine is that Herba Artemisiae Annuae and Radix Tripterygii Wilfordii two kinds of medical materials mix, then adopt supercritical extraction process to prepare extract; The weight ratio of described Herba Artemisiae Annuae and tripterygium wilfordii is 1:1;
The condition of described supercritical extraction is: CO
2flow is 4.6Lh
-1, extraction temperature 45 DEG C, extracting pressure 25MPa, employing ethanol is entrainer, and entrainer consumption is 2mLg
-1, extraction time 100min.
2. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine according to claim 1 is to the application of composition of medicine in preparation treatment rheumatoid arthritis agents.
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