CN103446220A - Sweet wormwood thunder god vine medicine pair combination drug and preparation method and application thereof - Google Patents
Sweet wormwood thunder god vine medicine pair combination drug and preparation method and application thereof Download PDFInfo
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- CN103446220A CN103446220A CN2013104514349A CN201310451434A CN103446220A CN 103446220 A CN103446220 A CN 103446220A CN 2013104514349 A CN2013104514349 A CN 2013104514349A CN 201310451434 A CN201310451434 A CN 201310451434A CN 103446220 A CN103446220 A CN 103446220A
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Abstract
The invention provides a sweet wormwood thunder god vine medicine pair combination drug. The sweet wormwood thunder god vine medicine pair combination drug is composed of a mixture or extractant of sweet wormwood and thunder god vine. The invention further provides the extractant of the sweet wormwood thunder god vine medicine pair combination drug. The extractant of the sweet wormwood thunder god vine medicine pair combination drug has lower toxicity for liver and kidney compared with the extractant of thunder god vine and has better activity for treating rheumatic arthritis compared with commercially available thunder god vine medicinal preparation.
Description
Technical field
The present invention relates to a kind of Chinese medicines to combination, relate in particular a kind of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to composition of medicine, preparation method and application.
Technical background
(Tripterygium wilfordii) is warm in nature for Radix Tripterygii Wilfordii, 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, antibiotic, pain relieving isoreactivity.The discovered in recent years Radix Tripterygii Wilfordii is evident in efficacy to aspects such as the rejection of organ transplantation, autoimmune disease, nephrotic syndrome, cancers, 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 is separated and is obtained about 7O kind composition from tripterygium plant, is mainly alkaloids, Diterpenes (as triptolide, i.e. Radix Tripterygii Wilfordii lactone alcohol), triterpenes, sesquiterpenoids and polysaccharide.Radix Tripterygii Wilfordii has been that one of maximum Chinese herbal medicine of poisoning occurs report since nearly over half a century.Along with the increase of people to 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.This paper is summarized the research of relevant Radix Tripterygii Wilfordii toxicity in recent years.
Tripterygium wilfordii Poisoning mainly be take cardiac muscle, nervous system, kidney damage as main, conscience even occurs hemorrhage or downright bad, and rapidly, mortality rate is high in development.Symptom is carrying out property and increases the weight of, first meeting is felt sick, vomiting, stomachache, diarrhoea etc., thready and rapid pulse appears then and weak, Blood pressure drop, giddy, headache, weak, nervous, irritated so that twitch, lumbago, oliguria, hematuria, albuminuria, non-protein nitrogen then may occur to raise, finally because of the list such as acute renal failure, circulatory failure, central nervous system's damaged nerve cell and serious bone marrow depression or multiple organs failure lethal.
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 very approaching, 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 the Radix Tripterygii Wilfordii dosage and being the effective thinking that reaches attenuation synergistic according to certain compatibility the form of the rules compound recipe.Animal and clinical experiment demonstration, arteannuin and derivant thereof have treats the RA effect preferably, and its toxicity is minimum.This research, by low dose of tripterygium glycosides and artesunate compatibility, has great importance for exploration safety, rheumatoid arthritis medicine efficient, low toxicity.
Summary of the invention
The huge toxicity problem existed for solving Tripterygium Preparations, the invention provides a kind of Herba Artemisiae Annuae (dry aerial parts that Herba Artemisiae Annuae of the present invention is Herba Artemisiae annuae Artemisia annua L.) and Radix Tripterygii Wilfordii medicine to composition of medicine, the mixture that described composition of medicine is Herba Artemisiae Annuae and two kinds of medical materials of Radix Tripterygii Wilfordii or its extract form.
Described composition of medicine is that Herba Artemisiae Annuae and two kinds of medical materials of Radix Tripterygii Wilfordii are mixed, 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 after Herba Artemisiae Annuae is mixed with tripterygium wilfordii, to adopt supercritical extraction to extract.
The preparation method of described Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to composition of medicine, the condition of described supercritical extraction is: CO
2flow is 4.6Lh
-1, 45 ℃ of extraction temperature, extracting pressure 25MPa, employing ethanol is entrainer, the entrainer consumption is 2mLg
-1, extraction time 100min.
Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine described above is the application in preparation treatment rheumatic arthritis medicine to composition of medicine.
Useful technique effect 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 active better in treatment rheumatic arthritis disease with commercially available Radix Tripterygii Wilfordii pharmaceutical preparation.
The specific embodiment
Embodiment 1 Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is investigated process for super-critical extracting
Take 5 parts, the Herba Artemisiae Annuae of 100.0g drying and tripterygium wilfordii (1:1) mixture, use respectively methanol, ethanol, 75% ethanol, ethylene glycol and ethyl acetate as entrainer, extracted, get a without entrainer in contrast;
Above-mentioned Herba Artemisiae Annuae and tripterygium wilfordii mixture are dosed in supercritical extraction reactor, and follow procedure heats up and pressurizes, and carries out dynamic extraction, CO
2flow is 4.6Lh
-1, 45 ℃ of extraction temperature, extracting pressure 25MPa, the entrainer consumption is 2mLg
-1, quantitatively pump into entrainer, extraction time 100min; Collect product from extraction-container, the reclaim under reduced pressure organic solvent, and obtain Herba Artemisiae Annuae tripterygium wilfordii supercritical extract, concrete outcome is as shown in table 1:
The impact of the different entrainers of table 1 on Radix Tripterygii Wilfordii lactone alcohol and artesunate extraction
As can be seen from Table 1, methanol, ethanol, 75% ethanol, ethylene glycol and five kinds of entrainers of ethyl acetate have all improved solute to a certain extent at SC-CO
2in dissolubility, but four kinds of different entrainers are to the increase rate difference of solid solute dissolubility, ethanol is the strongest and 75% ethanol is the strongest, methanol takes second place, ethylene glycol takes second place again, and the acetic acid ethyl dissolution amplitude is the most weak, and the association formed between this and entrainer and solute is 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, thereby improve Radix Tripterygii Wilfordii lactone alcohol (triptolide) and the dissolubility of artesunate isoreactivity composition in solvent; But, due to the polarity difference of five kinds of materials, 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.
SC-CO has been given in the introducing of entrainer
2the application that abstraction technique is more wide, also brought two negative effects simultaneously.Here it is due to the use of entrainer, increased the difficulty of Separation and Recovery entrainer from extract.And, owing to having used entrainer, make in some extracts the residual of entrainer arranged.This has just lost 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 impact of the different extracting method of embodiment 2 on the active component rate of transform
1, the preparation of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to ethanol extract
By 95% ethanol extraction 2 times for Herba Artemisiae Annuae and tripterygium wilfordii (1:1) mixture, amount of alcohol is 8 times of medical material amount. extraction time is respectively 2,1.5h, and 180 ℃ of boilings, 130 ℃ keep micro-boiling, and are respectively 2,1.5h.After having extracted, medicinal residues are poured out, dried.The extracted twice thing merges.Concentrating under reduced pressure 95% ethanol extraction, temperature is 50 ℃, and rotating speed is 34r/min, and pressure is-0.095MPa, is concentrated into 2L.Take out 500mL and keep sample, in water-bath evaporation as for, temperature is 82.5 ℃, then puts into baking oven and be dried into dry thing (temperature of baking oven is 42.5 ℃), obtains Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to alcohol extract.
2, the preparation of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to supercritical extract
Adopt the method described in embodiment 1, by the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:1), take medical material, pulverizing, adopt 75% ethanol as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to the supercritical extract I.
Adopt the method described in embodiment 1, by the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (3:1), take medical material, pulverizing, adopt 75% ethanol as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to the supercritical extract II.
Adopt the method described in embodiment 1, by the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:3), take medical material, pulverizing, adopt 75% ethanol as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to the supercritical extract III.
Adopt the method described in embodiment 1, by the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (4:1), take medical material, pulverizing, adopt 75% ethanol as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to the supercritical extract IV.
Adopt the method described in embodiment 1, by the ratio of Herba Artemisiae Annuae and tripterygium wilfordii (1:4), take medical material, pulverizing, adopt 75% ethanol as entrainer, prepares Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to the supercritical extract V.
3, the preparation of tripterygium wilfordii supercritical extract
Adopt the method described in embodiment 1, tripterygium wilfordii is pulverized, adopt 75% ethanol as entrainer, prepare the Radix Tripterygii Wilfordii supercritical extract.
3, the preparation of Artemisia annua supercritical extract
Adopt the method described in embodiment 1, by the Herba Artemisiae annuae pulverizing medicinal materials, adopt 75% ethanol as entrainer, prepare the Herba Artemisiae Annuae supercritical extract.
The lesions of liver and kidney effect of 4 pairs of mices of embodiment is investigated
The ICR mice, clean level, male and female half and half, weight 18-22g, totally 180, mice is divided into 9 groups at random, is respectively: the blank group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to the alcohol extract group; Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine is to the 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.Every group of laboratory animal in two batches, 10 every batch, 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 the blank group gives the equal-volume normal saline, and every day, gastric infusion was 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), two batches of animal eye sockets are got to blood lethal, in the centrifugal 10min of 3000r/min, get serum, mensuration ALT, AST and BUN, Scr level.
Application SPSSI6.0 statistical software.Measurement data with
mean.There is statistical significance P<0.05 for difference.
With the blank group, compare,
*p ﹤ 0.05,
*p ﹤ 0.01.
Table 2 result shows, after administration 5d, with the blank group, compare, 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 to supercritical extract V group and Radix Tripterygii Wilfordii supercritical extract group ALT and AST value to supercritical extract III group and Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine to alcohol extract group, Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, and and the blank group relatively there were significant differences (P<0.05 or 0.01).Analyze above data and find, when the ratio of Herba Artemisiae Annuae and Radix Tripterygii Wilfordii, during lower than 1:3, the ALT of 5d and 10d raises rapidly, and when 10d, AST also raises rapidly.
With the blank group, compare,
*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 the blank group relatively there were significant differences (P<0.05 or 0.01).But administration 5d is little on the Ser impact of mice.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 the blank group compares, and there were significant differences.Ethanol extract group Scr value raises, and blank group comparing difference remarkable (P<0.05).
Embodiment 4
Female Wistar rats, 130~170g.Test preposition animal in laboratory endoadaptation environment 3 days, room temperature 18-2O ℃, relative humidity 65%.Animal is divided into to 9 groups at random: 1. normal group, normally nursing; 2. model group, make the rat arthritis model that the II Collagen Type VI is induced.3. commercially available Tripterygium Hypoglaucum Hutch Tablet group (commercially available group), give commercially available Tripterygium Hypoglaucum Hutch Tablet (the accurate word Z20027411 of traditional Chinese medicines) 20mg/d after the rat arthritis model 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 the supercritical extract I after making, and dosage is the 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 the supercritical extract II after making, and dosage is the 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 the supercritical extract III after making, and dosage is the 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 the supercritical extract IV after making, and dosage is the 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 the supercritical extract V after making, and dosage is the 28g(crude drug)/kg.
The rat arthritis model that the II Collagen Type VI is induced: solubility in acid II Collagen Type VI 50mL is dissolved in 0.1mol, in the 25mL acetum, then 4 spend the night drips equal-volume complete Freund's adjuvant (CTA) and grinds and evenly make Emulsion to make collagen content be 1mg/mL under ice bath, inject respectively 0.1,0.2 at rat root of the tail section, back and cervical region Intradermal, 0.2mL collagen breast (0.5mL/ only), use ethanol partly sterilised before injection.The 14th day with 100 μ L/ the CTA containing collagen 1mg/mL only immune rat again.The rat hindleg of take occurs that redness is as the onset time, and each group is randomly drawed rat investigation indices and comprised the performance of inspection sign, local joint tissue pathological examination, immunologic test, hematological examination, image analysis.
The method that affects on the inflammation integration: experimental rat, after the immunity of secondary calf II Collagen Type VI, is observed respectively toes swelling situation, by the relevant requirements grade scale, keep the score, record once, until experiment finishes the previous day, adopts every rat integration addition method every other day.
The 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 and model group relatively have significant difference (P<0.05), heavy dose of group relatively has significant difference (P<0.O1) with model group, and the inflammatory effect of the autoimmune arthritis that GSF has remarkable inhibition II Collagen Type VI to bring out is described.Specifically as shown in table 4.
Table 4 inflammation integration record
Annotate: with model group, compare,
*p ﹤ 0.05,
*p ﹤ 0.01.
The II Collagen Type VI is induced to arthritis SOD in serum content influence method: undertaken by the test kit description.
The II Collagen Type VI is induced to arthritis SOD in serum content influence SOD content model group and Normal group there was no significant difference relatively in serum as a result, but alcohol extraction small dose group SOD in serum content is significantly higher than model group.Specifically as shown in table 5.
In sum, each is organized Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine supercritical extract is had antiinflammatory, regulates immune pharmacological action; Rheumatoid arthritis is had to clear and definite therapeutical effect.Wherein Herba Artemisiae Annuae and tripterygium wilfordii (1:1), Herba Artemisiae Annuae and two groups of medicines of tripterygium wilfordii (1:3) are best to the activity of compositions, Herba Artemisiae Annuae and tripterygium wilfordii (1:4) take second place, Herba Artemisiae Annuae and tripterygium wilfordii (3:1) Herba Artemisiae Annuae and two groups of medicines of tripterygium wilfordii (4:1) the poorest to the activity of compositions.From above-mentioned, can know, the ratio of Herba Artemisiae Annuae and tripterygium wilfordii is that 1:1-3 is optimal proportion.
Claims (7)
1. a Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine, to composition of medicine, is characterized in that: the mixture that described composition of medicine is Herba Artemisiae Annuae and two kinds of medical materials of Radix Tripterygii Wilfordii or its extract form.
2. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine according to claim 1, to composition of medicine, is characterized in that: described composition of medicine is that Herba Artemisiae Annuae and two kinds of medical materials of Radix Tripterygii Wilfordii are mixed, and then adopts process for super-critical extracting to prepare extract.
3. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine according to claim 1, to composition of medicine, is characterized in that: the weight ratio of described Herba Artemisiae Annuae and tripterygium wilfordii is 1:1-3.
4. Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine according to claim 3, to composition of medicine, is characterized in that: the weight ratio of described Herba Artemisiae Annuae and tripterygium wilfordii is 1:1.
5. the Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine preparation method to composition of medicine, it is characterized in that: described method is employing supercritical extraction extraction after Herba Artemisiae Annuae is mixed with tripterygium wilfordii.
6. the preparation method of Herba Artemisiae Annuae Radix Tripterygii Wilfordii medicine according to claim 3 to composition of medicine, it is characterized in that: the condition of described supercritical extraction is: CO
2flow is 4.6 Lh
-1, 45 ℃ of extraction temperature, extracting pressure 25MPa, employing ethanol is entrainer, the entrainer consumption is 2mLg
-1, extraction time 100min.
7. the described Herba Artemisiae Annuae Radix Tripterygii Wilfordii of claim 1-3 medicine application in preparation treatment rheumatic arthritis medicine to composition of medicine.
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CN1082914A (en) * | 1993-05-27 | 1994-03-02 | 李志铭 | Antirheumatic and manufacture method thereof |
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