CN101186627B - Method for preparing picroside I - Google Patents
Method for preparing picroside I Download PDFInfo
- Publication number
- CN101186627B CN101186627B CN2007101643215A CN200710164321A CN101186627B CN 101186627 B CN101186627 B CN 101186627B CN 2007101643215 A CN2007101643215 A CN 2007101643215A CN 200710164321 A CN200710164321 A CN 200710164321A CN 101186627 B CN101186627 B CN 101186627B
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- Prior art keywords
- kutkin
- injection
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- liver
- hepatitis
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Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention relates to the research and development technical field of new Chinese medicine, in particular to a preparation method of kutkoside I, which uses advanced extraction, separation and purification technique to extract the effective component kutkoside I from picrorhiza rhizome medicinal material for treating hepatitis B, while the content of the kutkoside I is higher than 90%. The inventive preparation method has low cost, simple operation and industrialization production suitability.
Description
Technical field
The present invention is dividing an application of 200410098646.4 applications for a patent for invention, and the original bill applying date is on December 15th, 2004, and the original bill denomination of invention is application and preparation and the preparation method of kutkin I in preparation treatment hepatitis B medicament.The invention belongs to new Chinese medicine research and development field, be specifically related to a kind of preparation method of kutkin I.
Background technology
As everyone knows, China is a hepatitis B diseases big country.At present, the whole world estimates at the medicine that is used to treat viral hepatitis more than 600 kinds, but does not still have gratifying medicine so far.World Health Organization West Pacific region eternal honor chairman doctor Han Xiangtai represent, current the most frequently used treatment hepatitis B medicament is cell Interferon, rabbit (Cytokine Interferon) and nucleoside analog (Nucleoside Analogues).Cell Interferon, rabbit commonly used comprises interferon-' alpha ', and Interferon, rabbit must be injected every day, and spinoff is more; Comprise flu symptom, myalgia, depression and diarrhoea etc. such as headache and fever occur; Whether medication also depends on liver function, if liver function is badly damaged, then is not suitable for using Interferon, rabbit.Another kind of cell Interferon, rabbit Peg-intron is also bringing into use.Nucleoside analog as for commonly used then comprises lamivudine (Lamivudine) and Adefovir (Adefovir).Lamivudine is used as the treatment chronic hepatitis B, belongs to oral pharmaceutical, few side effects, and the usefulness that suppresses virus is high, resistance can occur but take for a long time, produces the virus of mutation.The Adefovir toxic side effect is few, and this medicine also is used to suppress the research of HIV, but discovers that large dose oral administration can influence renal function, and renal function bad person should not use.Use cell Interferon, rabbit and one of shortcoming of examining the beanstalk analogue, be that all life-time service can cause severe side effect.Part is regulated for example Zadaxin (Thymosin) therapeutic efficiency that also come to light of immune medicine, but is not widely used as treating hepatitis B at present, because detailed therapeutic efficiency is clear not as good as other medicines, price is also quite expensive.
Up to the present, there is not a kind of medicine absolute utility to be arranged for suppressing hepatitis B.Doctor Han Xiangtai thinks, treats in future on the method for hepatitis B, and herbal medicine can be taken on an important role.The World Health Organization also with the treatment by Chinese herbs hepatitis B as newly leading future, and hope with the therapy as future blending the east and the west.Therefore people focus on sight on the Chinese herbal treatment hepatitis B gradually.
The habitual always in the past Chinese herbal treatment hepatitis B of the traditional Chinese medical science is used treatment by Chinese herbs hepatitis and has been experienced secular practice, has also accumulated rich experience.Chinese herbal treatment hepatitis B inexpensive, curative effect is reliable and spinoff is little, more and more becomes the main medicine of China's treatment hepatitis B.The treatment by Chinese herbs of hepatitis B comprises following several method: (1) clearing liver dampness removing promotes that jaundice disappears; (2) liver microcirculation that improves promoting blood circulation and removing blood stasis; (3) QI and blood regulating is transferred the general immunity function; (4) tonifying speen and tonifying kidney improves the clinical symptom of hepatitis; (5) various complication are treated in enriching yin, tonifying Qi.But herbal medicine all is being short of science data aspect validity of treating hepatitis B and the security., press for to numerous hepatitis B disease patients provides pharmacological action clear and definite, drug effect is high, the medicine of the treatment hepatitis B that toxic side effect is low for this reason.
The Chinese medicine Rhizoma Picrorhizae is former in Tang Materia Medica meaning its " Great Cold ", " main hectic fever due to yin labor heat, the tonifying spleen courage makes eye bright "." Xinxiu Bencao " say " main hectic fever due to yin labor heat, the tonifying liver courage makes eye bright ... thick stomach "." Kaibao Bencao " called its " bitter, flat, nontoxic ".In " the thunder fire is boiled in water for a while, then dress with soy, vinegar, etc. pharmacy property and separated ", it is returned through " going into liver, courage, stomach three warps ".MIAO Xi-Yong " Bencao Jingshu " say " Rhizoma Picrorhizae, kind remove damp and hot ... all are damp and hot, heat symptoms caused by an exopathgen, the cloudy latent heat institute that divides is sick all, there's no one who doesn't or isn't eliminates." " medicineization justice " say: " Rhizoma Picrorhizae is solely gone into blood system and heat-clearing." " wet fire knot gathers " book on Chinese herbal medicine justice " speech, non-this disease institute that can not go directly." Rhizoma Picrorhizae kind remove damp and hot, disappear and gather, the tonifying liver courage can be removed the damp and hot disease and evil of chronic hepatitis B, protects the liver courage helping except that the epidemic disease poison, for thorough this disease of treatment provides foundation.
Rhizoma Picrorhizae is a goatweed Rhizoma Picrorhizae Picrorhiza scrophulariifloraPennell dry rhizome; Be per nnial herb, grow on the rock of extremely frigid zones and in the mound, perhaps the area without shade of shallow soil layer; Be distributed in western Sichuan, northwestern Yunnan Province, southern Tibet.It is documented, contain iridoid glycosides, cucurbitacine, phenolic glycoside class in the Rhizoma Picrorhizae, also contain a spot of aromatic acid and D-N.F,USP MANNITOL in addition, wherein kutkin I (Picroside I) belongs to the iridoid glycoside compounds.The preparation method of relevant kutkin I and the application in the medicine of preparation treatment hepatitis B thereof also do not appear in the newspapers.
The inventor adopts Traditional Chinese medical theory and modern science achievement to combine; Adopt advanced extraction separation purification technique, from Chinese medicine Rhizoma Picrorhizae medicinal material, extract the effective constituent kutkin I of its treatment hepatitis B, make its content surpass 90%; And pharmacodynamics, pharmacological toxicology that kutkin I and corresponding pharmaceutical prepn thereof have carried out the treatment hepatitis B learned research; The result finds that this monomer pharmacological action is clear, has effects such as the hepatitis B replication of inhibition, protecting liver, lowering enzymes, promoting the function of the gallbladder to alleviate jaundice, immunomodulatory, and toxic side effect is extremely low; Safe to use, can be the new Chinese medicine that vast hepatitis B patient provides a kind of high-efficiency low-toxicity.
Summary of the invention
The object of the present invention is to provide the pharmaceutical dosage form of the preparation method and the treatment hepatitis B thereof of kutkin I.
The present invention further provides the Chinese medicine composition that is used to treat hepatitis B that contains kutkin I.
Kutkin I (Picroside I) belongs to the iridoid glycoside compounds.This compound is white unformed powder, soluble in water and methyl alcohol, fusing point 100-102 ℃.Molecular formula is C
24H
28O
11, structural formula is following:
The invention provides the preparation method of kutkin I with commercial value.Its separation purification method is quick, simple, practical, and production cost is lower, and suitability for industrialized production is highly advantageous to.
The method of kutkin I extraction provided by the invention, separation, purifying is following:
(1) extracts: dry rhizome or its pulverizing article of getting Tibet picrorhiza rhizome (Picrorhiza scrophulariiflora Pennell) and India's Rhizoma Picrorhizae (Picrorhiza kurroa Royle ex Benth); Water, ethanol, acetone equal solvent or its two kinds, the extraction of multiple mixed solvent; Extracting solution is being evaporated to dried (or organic solvent-free) below 70 ℃; Add water to suitable volume, the centrifugal deposition of removing gets supernatant;
(2) separate: supernatant passes through the good adsorption column of conventional processing, difference water and Different concentrations of alcohol wash-out, and collection contains the thin layer inspection and knows the elutriant that kutkin I is arranged, concentrating under reduced pressure, vacuum-drying below 70 ℃, pulverizing gets the kutkin I bullion;
(3) purifying: the kutkin I bullion is used anhydrous alcohol solution, filter, filtrating is admixed silica gel; Water-bath volatilizes solvent, loads on the silicagel column top, with the two mixed solvent wash-out of ETHYLE ACETATE, acetone or its; Collect effluent, drying under reduced pressure, pulverize the kutkin I sample.
Used Rhizoma Picrorhizae medicinal material can be the medicinal material of pure natural in the said extracted process, also can be the medicinal material of artificial propagation.The Rhizoma Picrorhizae medicinal material of employing artificial growth can guarantee the steady sources of medicinal material, and is quality controllable, and can reduce production costs.
Used Rhizoma Picrorhizae medicinal material can be ground into the granularity of 10～30 orders size in the said extracted process, can help the extraction of effective constituent.
Used solvent can be water, ethanol, acetone in the said extracted process, both can use separately, also can merge with two or more solvent and use.The process for extracting that adopts can use backflow, diacolation or apparatus,Soxhlet's to extract.
The filler of used filling adsorption column can be the resin of nonpolar or low-pole in the above-mentioned sepn process, like styrene type (comprising vinyl toluene type, ethyl styrene type), vinyl cyanide type resin etc., specifically can adopt D-101 type, ZTC type, AB-8 type etc.
Optimal conditions in said extracted, separation, the purge process can pass through L
9(3
4) orthogonal table carries out the quadrature screening experiment, is index with the content and the rate of transform of kutkin I, filters out optimum experimental condition.
Kutkin I content is more than 95% in the kutkin I sample that makes according to the method described above, and the rate of transform is more than 55%.The extraction result is stable, and good reproducibility can be used for large-scale production.
Kutkin I of the present invention during disease, can use separately when being used to treat hepatitis B, and the form of pharmaceutical composition that also can be through containing kutkin I is used.
Pharmaceutical composition provided by the invention can be by method known in the art preparation, but and administered through oral, hypogloeeis, through administrations such as skin, muscle or subcutaneous, mucocutaneous, urethra, vagina or veins.
The invention provides with the kutkin I is activeconstituents, is used to treat the pharmaceutical prepn and the related dose forms of hepatitis B.This pharmaceutical prepn is to be the effective active composition with above-mentioned kutkin I, and has comprised acceptable other auxiliary material components on the pharmaceutics, and wherein said kutkin I content surpasses 90%, preferably is higher than 95%.Pharmaceutical prepn of the present invention comprises oral preparation and non-intestinal drug delivery agent, and wherein oral preparation comprises capsule, oral liquid, chewing agent, pill, tablet, granule etc., and non-intestinal drug delivery agent comprises injection liquid formulation and freeze-dried powder injection type etc.Pharmaceutical prepn of the present invention also comprises local administration preparation, comprises creme, ointment, patch, sprays etc.Formulation of the present invention is not limited to this.
Available auxiliary type agent can be conventional filling agents such as starch, dextrin or Schardinger dextrins, sucrose, stearate when the preparation oral prepns.Can prepare through methods such as aseptic spraying drying, low-temperature vacuum drying, lyophilizes at the preparation lyophilized injectable powder.Each preparation later stage preparation technology and equipment all belong to the routine techniques of pharmacy field, the present invention does not limit this, so will not detail at this.
Creativeness of the present invention is to prove the effect that kutkin I has the obvious treatment hepatitis B through serial experiment.In order to prove that kutkin I has therapeutic action to hepatitis B; Comprise therapeutic action acute, chronic hepatitis B; Kutkin I and related dose forms that the contriver uses the above method to make; Adopted the different animal model, administered through oral and injecting pathway have carried out a large amount of experimentation on animalies, and experimental result shows: (1) causes the liver of Rats with Acute Liver Injury and mouse that tangible prevention protection and therapeutic action are arranged to tetracol phenixin, D-amido semi-lactosi and PARACETAMOL BP98; (2) cause the liver of chronic hepatic injury rat that tangible prevention protection and therapeutic action are arranged to autoallergic and tetracol phenixin; (3) can obviously increase bilirubin direct content in the normal rat bile, tangible choleretic effect is promptly arranged; (4) can increase how bilirubin direct (TBIL), gpt (ALT) content in ester (ANIT) the icterogenicity liver injury mice serum of isothiocyanic acid-1-, promptly how ester causes mouse jaundice liver damage that the obvious treatment effect is arranged to isothiocyanic acid-1-; (5) can obviously increase normal mouse macrophage phagocytic index, non-specific immune function is had tangible enhancement; (6) can obviously be increased in the normal mouse immune organ weight, promote the generation of hemolysin, have the effect that strengthens humoral immune function; (7) the duck hepatitis B there is the obvious treatment effect; (8) can obviously reduce HBsAg and the antigenic content of e in the 2.2.15 cell culture supernatant.
Main pharmacodynamics experimental technique of the present invention and result are following:
Test an injection kutkin I and D-Gal is caused the therapeutic action of chmice acute liver injury
The injection kutkin I causes the chmice acute liver injury to D-Gal (D-Galn) provide protection is observed in variation through detecting mice serum ALT, AST.Get Kunming mouse by body weight be divided at random blank group, model control group, Glycyrrhizic acid,diammonium salt group (50mg/kg), injection Rhizoma Picrorhizae sweet-I little (5mg/kg), in (10mg/kg), big (15mg/kg) three dose groups; Except that the blank group; All the other D-Gals of respectively organizing equal abdominal injection 1000mg/kg cause the chmice acute liver injury; Each administration group 1h after modeling, 11h and 21h press 0.1mL/10g volume intravenous administration, behind the modeling 22h; Pluck eyeball and get blood survey Serum ALT, AST value, and calculate the liver coefficient.The result sees the following form.
Table 1 injection kutkin I causes the influence (
) of acute liver damage mice serum ALT, AST to D-Gal
Annotate: compare * P＜0.05, * * P＜0.01 with model control group; Compare △ △ △ P＜0.001 with the blank group.
Table 2 injection kutkin I causes the influence (
) of acute liver damage mouse liver coefficient to D-Gal
Annotate: compare △ P＜0.05 with the blank group.
Above-mentioned experimental result shows: the injection kutkin I is little, in, big three dose groups compare with model control group, ALT, AST value all have remarkable reduction (P＜0.05 or P＜0.01).
Conclusion: the injection kutkin I has the obvious treatment effect to the chmice acute liver injury that D-Gal brings out.
Test two injection kutkin Is to CCl
4Cause the prevention provide protection of rat chronic liver injury
Observe the injection kutkin I to CCl
4Cause the prevention provide protection of rat chronic liver injury.Get Wistar rat male and female half and half, according to body weight be divided at random blank group, model control group, Glycyrrhizic acid,diammonium salt group (30mg/kg), injection kutkin I little (3.5mg/kg), in (7mg/kg) and big (10mg/kg) dose groups.The equal tail vein injection administration of each administration group, every day 1 time, 4 weeks of successive administration.After the administration 7 days, except that blank group rat, the equal subcutaneous injection 25%CCl of all the other each rats
4Peanut oil solution 2mL/kg 2 times weekly, in totally 3 weeks, causes the chronic hepatic injury model.24 hours abdominal aortic bloods after the last administration are measured Serum ALT, AST, TP, ALB, MDA, SOD, GSH content and liver starch, liver oxyproline.The result sees the following form.
Table 3 injection kutkin I is to CCl
4Cause rat chronic liver injury serum AST, ALT influence (
)
Annotate: compare with model control group
*P＜0.05,
*P＜0.01,
* *P＜0.001; Compare with the blank group
△ △ △ P＜0.001.
Table 4 injection kutkin I is to CCl
4Cause the influence that the rat chronic liver decreases TP, ALB (
)
Annotate: compare with model control group
*P＜0.05,
*P＜0.01,
* *P＜0.001; Compare with the blank group
△ △P＜0.01.
Table 5 injection kutkin I is to CCl
4Cause the liver starch and the liver oxyproline of rat chronic liver injury influence (
)
Annotate: compare with model control group
*P＜0.05,
*P＜0.01; Compare with the blank group
△ △ △P＜0.001.
Table 6 injection kutkin I is to CCl
4Cause MDA, SOD, GSH content in the rat chronic liver injury serum influence (
)
Annotate: compare with model control group
*P＜0.05,
*P＜0.01,
* *P＜0.001; Compare with the blank group
△P＜0.05,
△ △ △P＜0.001.
Above-mentioned experimental result shows: the injection kutkin I can obviously reduce Serum ALT, AST, MDA and liver oxyproline, and increased SOD, GSH, TP, ALB and glycogen initial value have significant difference (P＜0.05, P＜0.01 or P＜0.001).
Conclusion: the injection kutkin I is to CCl
4Cause the rat chronic liver injury tangible prevention provide protection is arranged.
Test three kutkin Is and tetracol phenixin is caused the therapeutic action of rat chronic liver injury
Observe the kutkin I oral administration causes the rat chronic liver injury to tetracol phenixin therapeutic action.Get the Wistar rat and adopt tetracol phenixin (CCl
4) cause chronic hepatic injury after, be divided at random model control group, Biphenylylmethylcarbinol group (150mg/kg), kutkin I little (15mg/kg), in (30mg/kg), big (60mg/kg) dose groups, and set up blank group parallel running.Each organizes continuous gastric infusion, 4 weeks of successive administration.Blank group and model control group give equal-volume saline water.After the last administration 24 hours, carry out abdominal aortic blood after each treated animal anesthesia, separation of serum is surveyed Serum ALT, AST, TP, ALB value, and gets liver and do pathological tissue inspection, mensuration liver oxyproline and liver glycogen content.The result sees the following form.
Table 7 kutkin I is to the influence (
) of chronic hepatic injury rat blood serum ALT, AST, A/G
Annotate: compare with the blank group,
△ △ △P＜0.001; Compare with model control group,
*P＜0.05
*P＜0.01
* *P＜0.001.
Table 8 kutkin I is to the influence (
) of chronic hepatic injury rats'liver glycogen, hydroxyproline content and liver coefficient
Annotate: compare with the blank group,
△ △P＜0.01
△ △ △P＜0.001; Compare with model control group,
*P＜0.05
*P＜0.01
* *P＜0.001.
Above-mentioned experimental result shows: kutkin I is little, in, heavy dose of group and model control group relatively, ALT, liver oxyproline obviously reduce (P＜0.05), liver glycogen obviously raise (P＜0.05); In the kutkin I, heavy dose of group obviously reduces with model control group comparison AST, liver coefficient, A/G ratio obviously raise (P＜0.05).Pathological examination results each administration group of prompting and model control group relatively do not have significant difference.
Conclusion: kutkin I has the obvious treatment effect to rat chronic liver injury due to the tetracol phenixin.
Test of the influence of four injection kutkin Is to normal macrophage phagocytosis of mice
Observe of the influence of injection kutkin I to normal macrophage phagocytosis of mice.Divide blank group, endoxan group (50mg/kg), LEVAMISOLE HCL group (30mmg/kg), injection kutkin I small dose group (5mg/kg), middle dose groups (10mg/kg), heavy dose of group (15mg/kg) at random with the ICR mouse; Totally 6 groups; The endoxan group is pressed the 0.02mL/10g intraperitoneal injection every day; The LEVAMISOLE HCL group is pressed the 0.2mL/10g gastric infusion, and each dose groups of injection kutkin I is all by 0.1mL/10g volume tail vein injection relative medicine, blank group tail vein injection equal-volume saline water; Once a day, continuous 7 days.24h after the last administration carries out carbon and cleans up experiment, calculates phagocytic index.Get Mouse Liver simultaneously and spleen is weighed, calculate organ coefficient.The result sees the following form:
Table 9 injection kutkin I is to the influence (
) of normal mouse macrophage phagocytic index
Annotate: compare △ P＜0.05 △ △ P＜0.01 with the blank group
Table 10 injection kutkin I is to the influence (
) of laboratory animal organ coefficient
Annotate: compare △ △ △ P＜0.001 with the blank group
Above-mentioned experimental result shows: in the injection kutkin I, heavy dose of group and blank group relatively, phagocytic index all rises to some extent, all has significance (P＜0.05) through statistical procedures difference, organ coefficient does not then have considerable change (P＞0.05).
Conclusion: the injection kutkin I has tangible enhancement to non-specific immune function.
Test of the influence of five injection kutkin Is to normal mouse humoral immune function
Through observing the injection kutkin I, detect its influence to humoral immune function to the influence that the normal mouse hemolysin generates.ICR kind mouse is divided into 6 groups at random by body weight; Be the blank group, LEVAMISOLE HCL (30mg/kg) group, endoxan group (20mg/kg) group; Injection kutkin I little (5mg/kg), in (10mg/kg), big (15mg/kg) dose groups; Wherein except that the LEVAMISOLE HCL group was pressed the 0.2mL/10g gastric infusion, each was organized all by 0.1mL/10g tail vein injection relative medicine, continuous 8 days.Administration second day, each organizes mouse peritoneal injection CRBC suspension 0.2mL, and continuous 7 days, 1h after the last immunity got blood and carries out hemolysin O D pH-value determination pH, got thymus gland, organ coefficient is weighed, calculated to spleen.The result sees the following form.
Table 11 injection kutkin I is to the influence (
) of normal mouse immune organ
Annotate: compare △ △ P＜0.01, △ △ △ P＜0.001 with the blank group.
Table 12 injection kutkin I is to the influence (
) of normal mouse hemolysin content
Annotate: compare △ P＜0.05 △ △ P＜0.01 △ △ △ P＜0.001 with the blank group
Above-mentioned experimental result shows: with the blank group relatively in the injection kutkin I, all obviously risings of heavy dose of group hemolysin O D value, through statistical procedures difference significance (P＜0.05, P＜0.001) is arranged all.Obviously raise with the heavy dose of group of blank group comparison injection kutkin I spleen coefficient, highly significant property (P＜0.01) is arranged through statistical procedures difference.
Conclusion: the injection kutkin I has the effect that strengthens the normal mouse humoral immune function.
Test the choleretic effect of six injection kutkin Is to normal rat
Through observing of the influence of injection kutkin I, the choleretic effect of research injection kutkin I to bilirubin direct content in choleresis and the bile.Method is got 50 of Wistar male rats; Body weight 220～240g; According to body weight be divided at random blank group, positive drug phenylpropyl alcohol capsule and pill group (250mg/kg), injection kutkin I little (3.5mg/kg), in (7mg/kg) and big (10mg/kg) dose groups, totally 5 groups.Open the abdominal cavity behind the rat anesthesia, make common bile duct intubated and drained bile, collect the preceding 0h of medicine～0.5h bile; Then respectively from sublingual vein administration 1mL/100g, collect 0h～0.5h after the administration, 0.5h～1.0h; 1.0h～1.5h; 1.5h～2.0h, 2.0h～2.5h, the bile flow of 2.5h～3.0h is also measured bilirubin direct content.The result sees the following form.
Table 13 injection kutkin I is to the influence (
) of rat bile flow
Annotate: compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with the blank group; With comparison △ P＜0.05 before the self administration, △ △ P＜0.01, △ △ △ P＜0.001.
Table 14 injection kutkin I is to the influence (
) of rat bile bilirubin direct
Annotate: compare * P＜0.05, * * P＜0.01, * * * P＜0.001 with the blank group; With comparison △ P＜0.05 before the self administration, △ △ P＜0.01, △ △ △ P＜0.001.
Above-mentioned experimental result shows: each dose groups of injection kutkin I is compared with the blank group, and each time period bile flow all has rising in various degree, and can obviously increase the bilirubin direct content (P＜0.05, P＜0.01, P＜0.001) in the bile.
Conclusion: the injection kutkin I has tangible choleretic effect.
Test seven injection kutkin Is to isothiocyanic acid-1-how ester cause the therapeutic action of mouse jaundice liver damage
The therapeutic action of injection kutkin I to the jaundice liver damage observed in variation through detecting mice serum total bilirubin (TBIL), conjugative bilirubin (DBIL), gpt (ALT), glutamic-oxal(o)acetic transaminase (AST).Kunming mouse by body weight be divided at random blank group, model control group, positive mattress Cape jasmine yellow (200mg/kg), injection kutkin I little (5mg/kg), in totally 6 groups of (10mg/kg), big (15mg/kg) dose groups, 12 every group.Except that the blank group, all the other each groups are all irritated clothes ANIT (100mg/kg) peanut oil solution by 0.1mL/10g and are caused chmice acute jaundice model, and 6h, 24h, 47h are administered three times after the modeling.After the last administration 1 hour (be after the modeling 48 hours), each treated animal is plucked eyeball and is got blood, and centrifugal serum is surveyed TBIL, DBIL, ALT, AST.Get liver, the calculating organ coefficient of weighing.The result sees the following form.
Table 15 injection kutkin I causes the influence (
) of mouse jaundice liver damage serum T BIL and DBIL to ANIT
Annotate: compare with model control group
*P＜0.05,
*P＜0.01; Compare with the blank group
△ △ △P＜0.001.
Table 16 injection kutkin I causes the influence (
) of mouse jaundice liver damage Serum ALT and AST to ANIT
Annotate: compare with model control group
*P＜0.05; Compare with the blank group
△ △ △P＜0.001.
Above-mentioned experimental result shows: in the injection kutkin I, heavy dose of group and model control group relatively, serum T BIL, ALT value be obviously reduction all, has significant difference (P＜0.05) through statistical procedures.All the other each indexs and model control group compare, no significant difference (P＞0.05).
Conclusion: the injection kutkin I has the obvious treatment effect to mouse jaundice liver damage.
Test of the influence of eight injection kutkin Is to the duck hepatitis B
Observe of the influence of injection kutkin I to DHB (DHBV).Experiment is established 6 groups; Normal control group, model group, positive drug group (lamivudine 20mg/kg), injection kutkin I little (10mg/kg), in (20mg/kg), big (40mg/kg) dose groups; Except that the normal control group; All the other each group all adopts sheldrake hepatitis B model, and intravenous route is adopted in administration, detects (T before the administration respectively
0), the 5th day (T after the administration
5), the 10th day (T
10) and drug withdrawal after the 3rd day (P
3) content of DHBV-DNA and preceding s surface antigen (DHBpresAg) in the duck serum.Three batches of experiment repetitions.The result sees the following form.
Table 17 injection kutkin I (HD-I) is to the influence of duck serum DHBV-DNA content
Annotate: *: t check in pairs: administration group different time (T
5, T
10, P
3) (T before duck serum DHBV-DNA average optical density value and the administration not
0) average optical density value relatively.*P＜0.05，**P＜0.01，***P＜0.001。
△: t check between group: administration group different time (T
0, T
5, T
10, P
3) duck serum DHBV-DNA average optical density value and model group (T
0, T
5, T
10, P
3) average optical density value relatively.△P＜0.05，△△P＜0.01，△△△P＜0.001
Table 19 injection kutkin I (HD-I) is to the influence of duck serum DHBpresAg level
Annotate: *: t check in pairs: administration different time (T
5, T
10, P
3) (T before duck serum DHBpresAg OD value and the administration not
0) the OD value relatively.*P＜0.05，**P＜0.01，***P＜0.001。
△: t check between group: administration group different time (T0, T5, T10, P3) duck serum DHBpresAg OD value and model group (T
0, T
5, T
10, P
3) the OD value relatively.△P＜0.05，△△P＜0.01，△△△P＜0.001。
Above-mentioned experimental result shows: to DHBV-DNA assay result in the duck serum be: in the three batches of experiments injection kutkin I little, in, significance descend (P＜0.001 is P＜0.01 perhaps) relatively arranged before the 3rd day duck serum DHBV-DNA average optical density value and the administration after the 5th day, the 10th day and the drug withdrawal after the administration of heavy dose of group; The injection kutkin I is little, in, after the administration of heavy dose of group the 5th day, the 10th day with drug withdrawal after the 3rd day serum DHBV-DNA level respectively with corresponding model group administration after the 5th day, the 10th day with drug withdrawal after compared in the 3rd day and all to have significance to descend (P＜0.001 or P＜0.01 or P＜0.05), and have certain dose-effect relationship.To DHBpresAg assay result in the duck serum be: in the three batches of experiments injection kutkin I little, in, significance descend (P＜0.001 perhaps P＜0.01 perhaps P＜0.05) relatively arranged before the 3rd day duck serum DHBpresAg OD value and the administration after the 5th day, the 10th day and the drug withdrawal after the administration of heavy dose of group; The injection kutkin I is little, in, after the administration of heavy dose of group the 5th day, the 10th day with drug withdrawal after the 3rd day serum DHBpresAg level respectively with corresponding model group administration after the 5th day, the 10th day with drug withdrawal after compared in the 3rd day significance descend (P＜0.001 or P＜0.01 or P＜0.05) all arranged.
Conclusion: in the injection kutkin I body dhbv dna is had than the obvious suppression effect.
Test nine injection kutkin Is to the inhibiting in vitro study of hepatitis B virus
Whether observation in vitro injection kutkin I is inhibited to hepatitis B virus (HBV).Detect the toxicity of injection kutkin I with mtt assay to the 2.2.15 cell; Detect the content that different concns injection kutkin I acts on behind the 2.2.15 cell different time HBV related antigen-surface antigen (HBsAg) and e antigen (HBeAg) in the cells and supernatant, the half toxic concentration (TC of calculating medicine pair cell with the ELISA method
50) and to antigenic half-inhibition concentration (IC
50), with TC
50And IC
50Ratio (being therapeutic index) estimate the validity and the security of medicine, TI＞2 are safe and effective.The normal control group is set simultaneously, blank group, and positive drug lamivudine (3TC) group parallel running.The result sees the following form.
Table 20 injection kutkin I is to the therapeutic index of HbsAg and HBsAg
Annotate: criterion is: TI＞2 are for effective; 1≤TI≤2 are poisonous effective; TI＜1 is a toxic action.
The explanation of above-mentioned experimental result: the three batches of experiment injection kutkin Is are respectively 2.97 and 4.83 to the MV of HBsAg and HBeAg therapeutic index; The positive drug lamivudine is 4.83 to the therapeutic index of HBsAg, and HBeAg is not had restraining effect.
Conclusion: the injection kutkin I is external to have the obvious suppression effect to HBV.
The pharmacodynamic experiment result shows that kutkin I not only has protecting liver, lowering enzymes, promoting the function of the gallbladder to alleviate jaundice and immunoregulation effect preferably, but also has significant anti-HBV effect.
In order to study the toxic action that kutkin I possibly exist body, estimate the drug safety that utilizes the Chinese medicine medicine that this monomer is prepared into, the inventor has carried out the general toxicity experiment and has tested with specific toxicity.
One, general toxicity experiment
1. studies on acute toxicity
Experimental result shows the LD of the injection type intravenous administration of kutkin I to mouse
50Value is 1040mg/kg, and intravenous administration is to the LD of rat
50Value is 171.4mg/kg, and intraperitoneal injection is to the LD of rat
50Be 552mg/kg.Medicine does not have obvious acute toxicity damage to vital tissue and the internal organs of animal.
2. long term toxicity research
(1) kutkin I injection type intraperitoneal injection is to the long term toxicity research of rat
Establish 4 groups altogether, be respectively solvent control group (equal-volume saline water), the little 40mg/ (kgd) of kutkin I injection type, middle 150mg/ (kgd) and big three dose groups of 300mg/ (kgd) are equivalent to 5 times, 21 times, 42 times of pharmacodynamics effective dose respectively.With general performance, body weight, food ration, hematology and serum biochemistry index, routine urinalysis, marrow classification, organ weights, organ coefficient before and after the animals administer serves as to detect index; And be aided with the pathological examination that animal is respectively organized internal organs, the toxicity situation of comprehensive evaluation medicine.The result shows that the heavy dose of treated animal body weight gain of kutkin I injection type is slow, and all the other each item indexs are all normal.In, each item of small dose group animal detects index and also do not see obviously unusual.The result of histopathologic examination shows that three dose groups animals are respectively organized the internal organs no abnormality seen.Be safe dose in can thinking below the dosage.
(2) kutkin I injection type intravenous drip administration is to the long term toxicity research of Beagle
Establish 4 groups altogether, be respectively solvent control group (equal-volume saline water), the little 20mg/ of kutkin I injection type (kgd), middle 77.46mg/ (kgd) and big three dose groups of 150mg/ (kgd).Be equivalent to 10,40 and 70 times of pharmacodynamics effective dose respectively.Observation index is the same.The result shows that large, medium and small dose groups animal each item detects also no abnormality seen of index.The pathology detection results suggest, three each vital tissue internal organs Non Apparent Abnormalities of dose groups animal.
Comprehensive above-mentioned two long term toxicity results of study, we think that injection kutkin I toxicity is lower, safety range is wide, considerably beyond the scope of pharmacodynamics effective dose.
Two, specific toxicity experiment
1. the mutagenesis of kutkin I research
Studied kutkin I potential mutagenicity in vitro and in vivo respectively through " Salmonella typhimurium reverse mutation experiment ", " Chinese hamster lung fibroblast (CHL) chromosome aberration experiment " and " mouse bone marrow cells is had a liking for the polychromatocyte micronucleus and formed experiment ".The result shows, no matter kutkin I in external still body, does not all have significantly direct or indirect mutagenesis.
2. the Study on Teratogenic of kutkin I
Sensitive period of teratogenesis reproductive toxicity test through kutkin I is observed kutkin I at rat sensitive period of teratogenesis successive administration, but whether the teratogenesis fetal hair is given birth to.The result shows that each dosage of kutkin I does not all have tangible teratogenesis.
In sum; The kutkin I toxic side effect is less; Pharmacological action is clear and definite, and it not only has protecting liver, lowering enzymes, promoting the function of the gallbladder to alleviate jaundice and immunoregulation effect preferably, but also has significant anti-HBV effect; This is one of creative place of the present invention, is the not available characteristics of domestic like product.As injection, this drug effect is rapid, and acute hepatitis is had good therapeutic action in addition.As oral preparation, its major advantage is less than toxicity such as Western medicine such as lamivudine, aciclovirs, should take for a long time.Above-mentioned experiment proof kutkin I is a kind of active compound that hepatitis B comprises the acute or chronic hepatitis B disease of treating, and the pharmaceutical composition that contains kutkin I is the good medicine of treating the hepatitis B disease simply.
Rhizoma Picrorhizae can adopt the method for artificial growth to breed at present, has guaranteed the steady sources of medicinal material, and is quality controllable, and a large amount of plantation can reduce production costs with exploitation.In view of China is hepatitis B big country, the medicine that kutkin I is developed to the treatment hepatitis B will have than wide market application prospect.
Specific embodiment
With following embodiment the present invention is done further elaboration, but the present invention is not limited to the content that the following example comprises.
The preparation method of embodiment 1 kutkin I
Get Rhizoma Picrorhizae medicinal material 3000g (kutkin I content 3.19% in the medicinal material), be ground into the medicinal powder of 10 mesh sieves.With 70% alcohol reflux of 12 times of medicinal material bulking values (v/w), it is 1.18～1.20 liquid concentrator that the gained percolate is condensed into specific density at 60 ℃, and the gained liquid concentrator adds the entry dilution, and is centrifugal; The gained supernatant with the speed of 1.2L/Kg.h through D-101 macroporous adsorptive resins (resin path: high=1: 10, resin demand is 2.5 times of medicinal material weight), treat that soup flows into resin fully after, use water, 32 times of medicinal material bulking values (v/w), the 15% ethanol elution impurity of 20 times of medicinal material bulking values (v/w) successively; Use 20 times of medicinal material bulking values (v/w), 25% ethanol elution kutkin I then, collect elutriant, 60 ℃ of concentrating under reduced pressure; Vacuum-drying is pulverized, and with absolute ethyl alcohol medicinal powder is dissolved again; Filter, filtrating is admixed silica gel, and 60 ℃ of water-baths volatilize solvent; Load on silicagel column top (the silica gel footpath: high=1: 10, the silica gel consumption is 1 times of medicinal material weight), with 10 times of medicinal material bulking values (V/W) eluent ethyl acetate; Collect effluent, 60 ℃ of drying under reduced pressure, pulverize kutkin I sample 56.85g; Kutkin I content 95.24%, kutkin I weight 54.14g, the kutkin I rate of transform 5 6.5 7%.(annotate: the volume number of one hour effluent of " L/Kg.h " expression per kilogram medicinal material, down together.)
The preparation method of embodiment 2 kutkin Is
Get Rhizoma Picrorhizae medicinal material 84kg (kutkin I content 3.19% in the medicinal material), be ground into the medicinal powder of 10 mesh sieves.With 70% ethanol speed diacolation of 8 times of medicinal material bulking values (v/w), it is 1.18～1.20 liquid concentrator that the gained percolate is condensed into specific density at 60 ℃, and gained liquid concentrator thin up is centrifugal; The gained supernatant is through ZTC type resin column, treat that soup flows into resin fully after, use water, 25 times of medicinal material bulking values (v/w), the 15% ethanol elution impurity of 18 times of medicinal material bulking values (v/w) successively, use 25 times of medicinal material bulking values (v/w), 25% ethanol elution then; Collect elutriant, 60 ℃ of concentrating under reduced pressure, vacuum-drying is pulverized; Use 1 times of medicinal material bulking value (v/w) absolute ethyl alcohol that medicinal powder is dissolved again, filter, filtrating is admixed silica gel; 60 ℃ of water-baths volatilize solvent, load on the silicagel column top, with 10 times of medicinal material bulking values (V/W) acetone wash-out; Collect effluent, 60 ℃ of drying under reduced pressure, pulverize kutkin I sample 1.55kg; Kutkin I content 95.30%, kutkin I weight 1.48kg, the kutkin I rate of transform 55.12%.
[embodiment 3] injection kutkin I preparation technology
Prescription: kutkin I 60.0g, process 1000.
Method for making: get kutkin I 60.0g, add the about 2800mL of water for injection, 65 ℃ of water-baths make dissolving, add total amount of preparation 2.0 ‰ gacs (6.0g); Stir half a hour, filter, on filter, add water to 3000mL, continue with 0.45 μ m, 0.22 μ m millipore filtration classified filtering; Filtrating quantitatively is sub-packed in the 10mL cillin bottle with every bottle of 3mL, and lyophilize recharges high-purity nitrogen, jumps a queue; Roll lid, packing promptly gets.
[embodiment 4] kutkin I tablet producing technology
Prescription: process 1000 altogether, each component and content are following:
Coptis glycosides I 30g
Starch 180g
Microcrystalline Cellulose 60g
Lactose 30g
Magnesium Stearate 1.5g
Method for making: get the kutkin I sample, wherein kutkin I content surpasses 90%, and it was pulverized 80 mesh sieves, mixes with starch, Microcrystalline Cellulose, lactose etc., with 8% starch slurry system softwood; Drying, whole grain adds Magnesium Stearate, mixing; Compressing tablet, quality inspection, packing promptly gets.
Claims (2)
1. the preparation method of a kutkin I is characterized in that:
(1) extracts: dry rhizome or its pulverizing article of getting Tibet picrorhiza rhizome (Picrorhiza scrophulariiflora Pennell) or India's Rhizoma Picrorhizae (Picrorhiza kurroa Royle ex Benth); Water, ethanol, acetone solvent or its two kinds, the extraction of multiple mixed solvent; Extracting solution be evaporated to below 70 ℃ dried; Add water to suitable volume, the centrifugal deposition of removing gets supernatant;
(2) separate: the supernatant of gained is passed through the good adsorption column of conventional processing; Used resin is D-101 type, ZTC type, AB-8 type water and Different concentrations of alcohol wash-out respectively; Collection contains the thin layer inspection and knows the elutriant that kutkin I is arranged; Concentrating under reduced pressure, vacuum-drying, pulverizing below 70 ℃ get the kutkin I bullion;
(3) purifying: the kutkin I bullion is used anhydrous alcohol solution, filter, filtrating is admixed silica gel; Water-bath volatilizes solvent, loads on the silicagel column top, with the two mixed solvent wash-out of ETHYLE ACETATE, acetone or its; Collect effluent; Drying under reduced pressure, pulverize the kutkin I sample, kutkin I content surpasses 90%.
2. preparation method according to claim 1 is characterized in that process for extracting is backflow, diacolation or apparatus,Soxhlet's process for extracting.
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| CN102920831B (en) * | 2012-11-20 | 2014-07-23 | 济南康众医药科技开发有限公司 | Preparation method of rhizoma picrorhizae total saponins |
| CN103601769B (en) * | 2013-11-27 | 2015-08-19 | 扬子江药业集团有限公司 | A kind of preparation technology of extraction separation and purification picroside Ⅰ and Picroside II from Rhizoma Picrorhizae |
| CN104402950B (en) * | 2014-12-29 | 2017-06-16 | 济南东源生物医药技术有限公司 | A kind of picroside Ⅰ crystallization preparation method |
| CN104693253A (en) * | 2015-03-30 | 2015-06-10 | 济南东源生物医药技术有限公司 | Industrial preparation method for high-purity 6'-O-trans-cinnamyl catalpol |
| CN104892699A (en) * | 2015-05-05 | 2015-09-09 | 济南东源生物医药技术有限公司 | Method for preparing high-purity 6'-O-trans-cinnamylcatalpol |
| CN105017355B (en) * | 2015-07-21 | 2017-10-24 | 济南同生生物医药科技有限公司 | The positive pressurized column chromatography preparation method of the trans cinnamyl Catalpols of 6 ' O |
| CN105198941A (en) * | 2015-09-21 | 2015-12-30 | 济南同生生物医药科技有限公司 | Aluminum oxide pressurized column chromatography preparation of 6'-O-trans cinnamyl catalpa alcohol |
| CN106749451B (en) * | 2017-03-27 | 2019-04-05 | 山东康裕生物科技有限公司 | A method of extracting separation picroside Ⅰ and picroside Ⅱ |
| CN112168834A (en) * | 2020-10-12 | 2021-01-05 | 上海中医药大学 | Application of picroside I in preparation of medicine for preventing and treating hepatic fibrosis |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5145955A (en) * | 1991-10-28 | 1992-09-08 | Council Of Scientific & Industrial Research | Process for the preparation and composition of a fraction containing picroside I and kutkoside |
| CN1458159A (en) * | 2002-05-15 | 2003-11-26 | 成都博瑞医药科技开发有限公司 | Process for preparing extract of picrorhiza iridoid glycoside |
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| US5145955A (en) * | 1991-10-28 | 1992-09-08 | Council Of Scientific & Industrial Research | Process for the preparation and composition of a fraction containing picroside I and kutkoside |
| CN1458159A (en) * | 2002-05-15 | 2003-11-26 | 成都博瑞医药科技开发有限公司 | Process for preparing extract of picrorhiza iridoid glycoside |
Non-Patent Citations (1)
| Title |
|---|
| 刘美凤.复方胡黄连抗肝炎有效部位的化学研究.黑龙江中医药大学硕士毕业论文.2002,31-45. * |
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