CN107875163A - A kind of Swertia mussotii active component for treating acute, chronic hepatitis and its preparation method and application - Google Patents
A kind of Swertia mussotii active component for treating acute, chronic hepatitis and its preparation method and application Download PDFInfo
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/12—Carboxylic acids; Salts or anhydrides thereof
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- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
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- A61K9/00—Medicinal preparations characterised by special physical form
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
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- A61K9/20—Pills, tablets, discs, rods
- A61K9/2004—Excipients; Inactive ingredients
- A61K9/2022—Organic macromolecular compounds
- A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
- A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
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- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- C07D—HETEROCYCLIC COMPOUNDS
- C07D407/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00
- C07D407/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings
- C07D407/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having oxygen atoms as the only ring hetero atoms, not provided for by group C07D405/00 containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/51—Gentianaceae (Gentian family)
Abstract
The invention discloses a kind of Swertia mussotii active component for treating acute, chronic hepatitis, the active component mainly contains four kinds of gentiamarin, sweroside, mangiferin and Swertiamarin active components, total content is 50~95%, wherein, the mass ratio of four kinds of active components is (40~90)：(1.0~10)：(0.5~5)：(0.5~5).Pharmaceutical composition the invention also discloses the preparation method of above-mentioned active component and comprising above-mentioned active component and its application in treatment acute, chronic hepatitis medicine is prepared.The Swertia mussotii active component of the present invention can significantly improve the hepatic injury of ANIT rats, can preferably treat acute, chronic hepatitis.
Description
Technical field
The invention belongs to middle Tibetan medicine technical field, and in particular to a kind of Swertia mussotii active component for treating acute, chronic hepatitis.
Background technology
Acute, chronic hepatitis is the liver cell destruction as caused by hepatitis viruse, hepatic tissue destruction reconstruct and timid pipe choking, is led
Cause with reference to bilirubin with unconjugated bilirubin to increase in blood, the jaundice of the part such as caused skin, mucous membrane and eyeball sclera
A kind of disease.Acute, chronic hepatitis can be caused by various hepatitis viruses, after hepatites virus infections, damage the choleresis device of liver cell
With the microvillus of bile capillaries, there is liver cell mitochondrial hypertrophy, smooth surfaced endoplasmic reticulum hyperplasia, golgiosome change in vacuolation
Become, lysosome increasing, the change such as bile capillaries microvillus thickening, rust, sparse or even disappearance, make choleresis and excretory function
A series of obstacles of generation, so as to cause cholestasis.The lesion of acute, chronic hepatitis is predominantly located at lobuli hepatis central part, capillary
Inside there is courage bolt, have BILE PIGMENTS calm in liver cell nuclear.There is inflammatory cell infiltration portal area, and its lesion degree is related to jaundice.
In terms of clinical treatment, commonly using Western medicine kind includes cortex hormone of aadrenaline, phenobarbital, deoxidation bear gall acid, Ademetionine etc.
Kind, curative effect is preferable, but also in the prevalence of adverse reactions such as lesions of liver and kidney, stomach reactions.Herbal species include compound cholagogic
Piece, compound dantong tablets, Yinzhihuang" granula etc., slow curative effect.
" Swertia mussotii " is that liver and bladder disease, hemopathic characteristic medicine resource plant are treated in Tibetan medicine medical science.Tibetan described in Tibetan medicine
Oriental wormwood is mainly 10 kinds of plants of number such as the Swertia of Gentianaceae, flower anchor category, Gentianopsis barbata category and Marsh Felwort category.Clinically commonly use and hide mattress
Old class medicinal material mainly includes mussot swertia herb, Swertia franchetiana H.Smith, Indian Herba Swertiae bimaculatae, Halenia elliptica D. Don etc..According to《Jingzhubencao》Deng
Document is recorded, and Swertia mussotii class medicinal material has the multiple efficacies such as clearing liver cholagogic, diuresis, be clinically widely used in cholestatic hepatitis,
The treatment of a variety of diseases of virus hepatitis, acute, chronic hepatitis, cholecystitis.Modern pharmacodynamic study proves, ring contained by " Swertia mussotii "
Alkene ether terpene and mouth mountain ketones component are that it treats the main pharmacodynamics composition of the hepatobiliary system disease such as hepatitis and cholecystitis.Cyclenes ether
Terpene is a kind of bitter compound, mainly including three kinds of Swertiamarin, gentiamarin and sweroside, has anti-liver
Poison, promote choleresis, anti-inflammatory and the effect such as antiviral；Mouth mountain ketone is the phenoloid of a kind of yellow, has solution liver
Poison, liver protection, anti-inflammatory, antiviral and suppression xanthine oxidase isoreactivity, mangiferin is wherein important one kind.In the last few years with
Swertia mussotii be primary raw material through modern crafts process prepare Tibetan medicine new drug have Jingzhu Gantaishu capsules, the base of a fruit up to ball, Swertia mussotii piece,
Swertia mussotii capsule, anxious proheparin, drug for curing B-hepatitis etc., it is applied to clinical mainly treating acute, chronic hepatitis, cholecystitis etc..Also pass
The Tibetan medicine kind of uniting include five tastes Swertia patens soup dissipates, eight taste swertia pills, eight taste Swertia patens dissipate, nine taste swertia pills, ten taste myrobalans
Soup etc..These Tibetan medicine kind drug effects are obvious, but effective substance is complicated, and its mechanism of action is not yet explained clear.
Chinese invention patent (the patent No.：200710061832.4) a kind of preparation technology of Tibet capilary artemisia is disclosed, should
Technique using Swertia mussotii as raw material, extracted through hydrophilic solvent, various hepatitis are treated in the retention method production of Gravity Separation and molecular sieve,
Cholecystitis, hepatic injury, liver fibrosis, the water injection and freeze dried powder of fatty liver diseases；Chinese invention patent (the patent No.：
200710061832.4) disclose it is a kind of by extracting, cleaning, concentrating, initial gross separation, adsorbing separation, concentration, drying, crush, enter
The extracting method for the Swertia mussotii active ingredient that the process equipments such as one one-step refining form, this method are extracted using water, ethanol, and ethanol surpasses
Sound extracts, using the absorption such as centrifugation, macroreticular resin, separating effective ingredient；Chinese invention patent (the patent No.：
200710163097.8) a kind of Tibetan capillary extract is disclosed, includes Swertiamarin, gentiamarin, sweroside, mango
5 kinds of glycosides, Lutonaretin compositions, total content 22.65-81.87%.The extract can improve indices caused by hepatic injury,
Promote the reverse of liver fibrosis process, preferably prevent hepatic sclerosis；Chinese invention patent (the patent No.：200910078578.8) public
A kind of Tibetan capillary extract is opened, wherein at least containing general flavone, general flavone accounts for the 20%-75% of the Tibetan capillary extract, the Tibetan
Capillary extract has the activity for the treatment of disease in the liver and gallbladder；Chinese invention patent (the patent No.：201010106232.7) disclose one
Kind contains iridoid, mouth mountain ketone and triterpenes components Tibetan capillary extract, and three kinds of composition total contents are 20%-80%.The extraction
Thing has obvious choleretic effect, has obvious guarantor to the dysfunction of liver of the hepatic injury of chemical substance induced Acute and obstructive jaundice
Shield and improvement result；Chinese invention patent (the patent No.：201210066770.7) disclose a kind of Swertia mussotii active component extraction
The preparation method of thing, including crushing, ultrasonic extraction, centrifugation clean, be concentrated under reduced pressure, ceramic membrane clarification, Ultra filtration membrane, macropore tree
The process such as fat purifying, dry.Iridoid and mouth mountain ketone content 15.5%-40.4% in extract.The active component of the present invention
Preparation method has advantage following prominent and good effect compared with existing patented technology：(1) this active component is in Swertia mussotii
It is further purified obtained on the basis of extract, active constituent content is high, and minimum content is more than more than 50%, up to the class of Chinese medicine five
The requirement of new drug, this is that conventional art is not reached；(2) this active component preparation process is not related in addition to using medical ethanol
And other such as petroleum ethers, chloroform, methanol, ethyl acetate, n-butanol are poisonous, inflammable chemical solvent, preparation technology is green；
(3) this active component preparation technology active ingredient concentrate, active component containing only Swertiamarin, gentiamarin, sweroside and
Four kinds of compositions of mangiferin, impurity is few, is easy to quality control；(4) this active component can significantly improve the hepatic injury of ANIT rats, and
Its therapeutic effect is significantly stronger than positive drug, and especially the most obvious with low dose group (0.04g/kg), the dosage reaches Isodose
The treatment level of chemicals.
The content of the invention
In order to solve problems of the prior art, the present invention chooses clinical effectively Tibetan medicine Swertia mussotii and makees medicinal material as research
Object, devise one kind using gentiamarin, sweroside, mangiferin and Swertiamarin as main component and treat acute and chronic liver
Scorching Swertia mussotii active component and preparation method thereof, pharmaceutical composition and application.It is an object of the invention to provide a kind of curative effect is true
Cut, the Swertia mussotii active component of quality controllable treatment acute, chronic hepatitis.
Another object of the present invention is to provide the preparation method of the Swertia mussotii active component of above-mentioned treatment acute, chronic hepatitis.
Still a further object of the present invention is to provide the drug regimen of the Swertia mussotii active component comprising above-mentioned treatment acute, chronic hepatitis
Thing.
The Swertia mussotii active component that the further object of the present invention is to provide above-mentioned treatment acute, chronic hepatitis is preparing treatment
Application in acute and chronic medicine, main pharmacodynamics act as significantly improving hepatic injury, removing jaundice and reduce transaminase effect.
A kind of Swertia mussotii active component for treating acute, chronic hepatitis, it is characterised in that contain in the Swertia mussotii active component
There are four kinds of gentiamarin, sweroside, mangiferin and Swertiamarin active components, total content is 50~95%, wherein, it is described
The mass ratio of four kinds of active components is (40~90)：(1.0~10)：(0.5~5)：(0.5~5).
Further, the gentiamarin, sweroside, the mass ratio of four kinds of active components of mangiferin and Swertiamarin are
80:10:3.2:1.8。
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis：
(1) extract：By Swertia mussotii pulverizing medicinal materials, with 1g：5mL~30mL ratio mass concentration is 0~95% second
Alcohol carries out circumfluence distillation at 50~90 DEG C, and extraction time is 1~3 time, and 1~3h, merging every time obtains extract solution；
(2) centrifugal treating：By 10000~20000rpm/min of extract solution high speed tubular type centrifuge, obtain
To supernatant；
(3) purifies and separates：Large pore resin absorption column on supernatant will be obtained, first the quality with 5~20 times of bed volumes is dense
The ethanol elution for 3~8% is spent, removes non-target components；It is again 10~20% with the mass concentration of 5~50 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：By above-mentioned active component eluent be concentrated under reduced pressure or with molecular cut off be 100~1000 organic film
Concentrated, obtain active component concentrate；
(5) dry：By above-mentioned active component concentrate depressurize oven drying or microwave drying or or freeze-drying or
Spray drying, dried object, which crushes, produces Swertia mussotii active component.
Further, the Swertia mussotii medicinal material be mussot swertia herb, it is Swertia franchetiana H.Smith, Indian Herba Swertiae bimaculatae, Swertia punicea Hemsl, ellipse
Roundleaf spends one or more of mixing in anchor or Marsh Felwort；
Further, the resin used is AB-8, D101, S-8, X-5, D3520, HPD100, HPD300 and HPD450A
In any one；
Further, the organic film is the organic film of resistance to alcohol；
Further, the condition that is concentrated under reduced pressure is 0.05~0.1Mpa of vacuum, 50~90 DEG C of temperature；
A kind of pharmaceutical composition for treating acute, chronic hepatitis, above-mentioned one kind comprising therapeutically effective amount treat acute, chronic hepatitis
Swertia mussotii active component, pharmaceutic adjuvant and pharmaceutical carrier.
Wherein, described pharmaceutical composition can be made into clinically acceptable formulation, as tablet, capsule, injection,
Any one in granula, pill, supensoid agent, dripping pill, oral liquid, aerosol, suppository or subcutaneous administration formulation.
A kind of described Swertia mussotii active component for treating acute, chronic hepatitis is in treatment acute, chronic hepatitis medicine is prepared
Using main pharmacodynamics act as significantly improving hepatic injury, removing jaundice and reduce transaminase effect.
Swertia mussotii active component provided by the invention, the active component HPLC assays for the treatment of acute, chronic hepatitis therein
Method is as follows：
Chromatographic condition and system suitability test：It is filler with octadecylsilane chemically bonded silica, Detection wavelength is
254nm。
Mobile phase：Methanol-water (0.1% acetic acid), gradient elution, 0-10min, 10%-40% methanol；10-30min,
40%-45% methanol.
The preparation of need testing solution：This product 20mg is taken, it is accurately weighed, it is placed in after being dissolved with methanol in 5mL volumetric flasks, methanol
Constant volume shakes up, produced in scale.
The preparation of reference substance solution：Precision weighs Swertiamarin standard items 5.4mg in 50mL volumetric flasks, adds methanol molten
Constant volume is solved, shakes up and produces 108 μ g/mL Swertiamarin standard solutions；Precision weighs gentiamarin standard items 5.5mg in 10mL
In volumetric flask, add methanol to dissolve constant volume, shake up and produce 550 μ g/mL gentiamarin standard solutions；Precision weighs sweroside mark
Quasi- product 5.3mg adds methanol to dissolve constant volume, shakes up and produce 106 μ g/mL sweroside standard solutions in 50mL volumetric flasks；Essence
The close mangiferin standard items 5.2mg that weighs adds 20% methanol to dissolve constant volume, shakes up and produce 104 μ g/mL mango in 50mL volumetric flasks
Glycosides standard solution.Swertiamarin, gentiamarin, sweroside and mangiferin singly mark solution 100,500,300 are drawn respectively
With 100 μ L, mix and produce mixed mark solution.
Assay method：It is accurate respectively to draw reference substance solution and each 10 μ L of need testing solution, high performance liquid chromatograph is injected,
Chromatogram is recorded, calculates, produces.
Compared with prior art, the invention has the advantages that：
(1) there is Swertia mussotii active component of the present invention obvious pharmacological effect to act on, and can significantly improve the liver of ANIT rats
Damage, and its therapeutic effect is significantly stronger than positive drug group (Swertia mussotii piece), and the liver organization form of normal rat is normal, nothing
Pathologic changes.
(2) preparation method of Swertia mussotii active component of the present invention, technique is simple, favorable reproducibility, is easy to operate and control, from
And be advantageous to large-scale industrial production, and prepare Swertia mussotii active component have the advantages that it is curative for effect, quality controllable,
Thus there is boundless application prospect.
Brief description of the drawings
Fig. 1 is gentiamarin, sweroside, the structure chart of four kinds of active components of mangiferin and Swertiamarin.
Fig. 2 is the HPLC chromatogram of reference substance solution, wherein, 1：Sweroside；2：Gentiamarin；3：Sweroside；4：
Mangiferin.
Fig. 3 is the HPLC chromatogram of active component, wherein, 1：Sweroside；2：Gentiamarin；3：Sweroside；4：Awns
Fruit glycosides.
The body weight of Fig. 4 rats is with the changing trend diagram of administration time (compared with blank：#P<0.05,##P<0.01,###P<
0.001)
Morphologic variation diagram (the A of rat in Fig. 5 experimentations:Blank group；B:Model group；C:Positive drug group；D:Low dosage
Group；E:Middle dose group；F:High dose group)
Fig. 6 rat liver variation diagrams (A:Blank group；B:Model group；C:Positive drug group；D:Low dose group；E:Middle dose group；
F:High dose group)
The liver weight of Fig. 7 each group rats and its corresponding liver index figure are (compared with model：#P<0.05,##P<0.01；
Compared with blank：*p<0.05,**P<0.01,)
The change figure of the drug-induced ANIT rat blood serums biochemical indicators of Fig. 8 is (compared with model：#P<0.05,##P<0.01；
The * p compared with blank<0.05,**P<0.01)
The impact effect figure one for the acute, chronic hepatitis liver tissues of rats morphological change that Fig. 9 medicines are induced ANIT
The impact effect figure two for the acute, chronic hepatitis liver tissues of rats morphological change that Figure 10 medicines are induced ANIT
Embodiment
According to following embodiments, the present invention may be better understood.It is however, as it will be easily appreciated by one skilled in the art that real
Apply and be merely to illustrate the present invention described by example, without should be also without limitation on this hair described in detail in claims
It is bright.
Embodiment 1
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis, comprises the following steps,
(1) extract：By oval leaf+report stem Swertia patens (1：1) pulverizing medicinal materials, with 1g：5mL ratio deionized water is 50
Circumfluence distillation is carried out at DEG C, extraction time is 1 time, extracts 1h, obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 20000rpm under (1) item, obtain
Clear liquid；
(3) isolate and purify：By S-8 type large pore resin absorption columns on the supernatant under (2) item, first with 5 times of bed volumes
Mass concentration be 3% ethanol elution, remove non-target components；The second for being again 10% with the mass concentration of 5 times of bed volumes
Alcohol elutes, and obtains active component eluent；
(4) concentrate：The active component eluent under (3) item is concentrated under reduced pressure to give active component concentrate；
(5) dry：The active component concentrate under (4) item is dried with microwave drying mode, gained dried object crushes
Produce Swertia mussotii active component.Tibetan capillary extract manufactured in the present embodiment, knot are determined using the foregoing HPLC methods of the present invention
Fruit is as follows：Gentiamarin, sweroside, the content of 4 main compounds of mangiferin and Swertiamarin account for active component solid
The 50% of thing content, the mass ratio of four kinds of compositions is 40:4.5:0.5:5.
Embodiment 2
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By Swertia franchetiana H.Smith pulverizing medicinal materials, with 1g：The ethanol that 30mL ratio mass concentration is 95% exists
Circumfluence distillation is carried out at 90 DEG C, is extracted 3 times, each 3h, merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 10000rpm under (1) item, obtain
Clear liquid；
(3) isolate and purify：By the D101 type large pore resin absorption columns under (2) item on the supernatant, first with 20 times of post beds
The mass concentration of volume is 4% ethanol elution, removes non-target components；It is again 14% with the mass concentration of 20 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：The active component eluent under (3) item is concentrated under reduced pressure to give active component concentrate；
(5) dry：By the decompression oven drying of the active component concentrate under (4) item, gained dried object, which crushes, to be produced
Swertia mussotii active component.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, as a result such as
Under：Gentiamarin, the content of 4 main compounds of sweroside mangiferin and Swertiamarin account for active component solid content
55%, the mass ratioes of four kinds of compositions is 44:4:5:2.
Embodiment 3
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By mussot swertia herb pulverizing medicinal materials, with 1g：The ethanol that 10mL ratio mass concentration is 10% exists
Circumfluence distillation is carried out at 70 DEG C, extraction time is 2 times, each 2h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 15000rpm under 3.1 (1) items, obtain
Supernatant；
(3) isolate and purify：By D3520 type large pore resin absorption columns on the supernatant under (2) item, first with 8 times of post bed bodies
Long-pending mass concentration is 8% ethanol elution, removes non-target components；It is again 20% with the mass concentration of 20 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：The active component eluent under (3) item is carried out with the organic film of resistance to alcohol that molecular cut off is 100
Concentration, obtains active component concentrate；
(5) dry：The active component concentrate under (4) item is dried with microwave drying oven, gained dried object, which crushes, is
Obtain Swertia mussotii active component.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, as a result
It is as follows：Gentiamarin, the content of 4 main compounds of sweroside mangiferin and Swertiamarin account for active component solid content and contained
The 57.5% of amount, the mass ratio of four kinds of compositions is 50:5:1:1.5.
Embodiment 4
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By Indian Herba Swertiae bimaculatae pulverizing medicinal materials, with 1g:The ethanol that 15mL ratio mass concentration is 30% exists
Circumfluence distillation is carried out at 70 DEG C, extraction time is 2 times, each 2h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 20000rpm/min under (1) item, obtain
To supernatant；
(3) isolate and purify：By X-5 type large pore resin absorption columns on the supernatant under (2) item, first with 10 times of bed volumes
Mass concentration be 6% ethanol elution, remove non-target components；The second for being again 17% with the mass concentration of 30 times of bed volumes
Alcohol elutes, and obtains active component eluent；
(4) concentrate：The active component eluent under (3) item is carried out with the organic film of resistance to alcohol that molecular cut off is 300
Concentration, obtains active component concentrate；
(5) dry：The active component concentrate under (4) item is dried with spray drying device, gained dried object crushes
Produce Swertia mussotii active component.Tibetan capillary extract manufactured in the present embodiment, knot are determined using the foregoing HPLC methods of the present invention
Fruit is as follows：Gentiamarin, the content of 4 main compounds of sweroside mangiferin and Swertiamarin account for active component solid content
The 60.4% of content, the mass ratio of four kinds of compositions is 51.5:6.2:1.5:1.2.
Embodiment 5
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：Mussot swertia herb pulverizing medicinal materials, with 1g：The ethanol that 20mL ratio mass concentration is 10% is 90
Circumfluence distillation is carried out at DEG C, extraction time is 3 times, each 2h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 20000rpm under (1) item, obtain
Clear liquid；
(3) isolate and purify：By AB-8 type large pore resin absorption columns on the supernatant under (2) item, first with 8 times of bed volumes
Mass concentration be 5% ethanol elution, remove non-target components；The second for being again 15% with the mass concentration of 50 times of bed volumes
Alcohol elutes, and obtains active component eluent；
(4) concentrate：The active component eluent under (3) item is concentrated under reduced pressure to give active component concentrate；
(5) dry：The active component concentrate under (4) item is freezed with freeze drier, gained dried object, which crushes, is
Obtain Swertia mussotii active component.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, as a result
It is as follows：Gentiamarin, the content of 4 main compounds of sweroside mangiferin and Swertiamarin account for active component solid content and contained
The 95% of amount, the mass ratio of four kinds of compositions is 80:10:3.2:1.8.
Embodiment 6
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By Indian Herba Swertiae bimaculatae pulverizing medicinal materials, with 1g：The ethanol that 25mL ratio mass concentration is 20% exists
Circumfluence distillation is carried out at 80 DEG C, extraction time is 3 times, each 2h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 10000rpm/min under (1) item, obtain
To supernatant；
(3) isolate and purify：By HPD100 type large pore resin absorption columns on the supernatant under (2) item, first with 10 times of post beds
The mass concentration of volume is 7% ethanol elution, removes non-target components；It is again 18% with the mass concentration of 20 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：The active component eluent under (3) item is carried out with the organic film of resistance to alcohol that molecular cut off is 1000
Concentration, obtains active component concentrate；
(5) dry：By the decompression oven drying of the active component concentrate under (4) item, gained dried object produces Tibetan mattress
Old active component.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, it is as a result as follows：Dragon
Courage hardship glycosides, the content of 4 main compounds of sweroside mangiferin and Swertiamarin account for active component solid content
81.4%, the mass ratio of four kinds of compositions is 76:3:0.6:0.8.
Embodiment 7
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By mussot swertia herb+report stem Swertia patens (1:1) pulverizing medicinal materials are mixed, with 1g：20mL ratio matter
The ethanol that amount concentration is 5% carries out circumfluence distillation at 90 DEG C, and extraction time is 3 times, each 2h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 10000rpm under (1) item, obtain
Clear liquid；
(3) isolate and purify：By HPD300 type large pore resin absorption columns on the supernatant under (2) item, first with 8 times of post bed bodies
Long-pending mass concentration is 4% ethanol elution, removes non-target components；It is again 10% with the mass concentration of 25 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：The active component eluent under (3) item is concentrated under reduced pressure to give active component concentrate；
(5) dry：The active component concentrate under (4) item is freezed with freeze drier, produces the effective portion of Swertia mussotii
Position.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, it is as a result as follows：Gentiamarin,
The content of 4 main compounds of sweroside mangiferin and Swertiamarin accounts for the 72.3% of active component solid content, four
The mass ratio of kind composition is 64:6:1.1:1.2.
Embodiment 8
A kind of preparation method for the Swertia mussotii active component for treating acute, chronic hepatitis
(1) extract：By mussot swertia herb+Halenia elliptica D. Don pulverizing medicinal materials, with 1g：10mL ratio is with mass concentration
10% ethanol carries out circumfluence distillation at 80 DEG C, and extraction time is 2 times, each 3h, and merging obtains extract solution；
(2) centrifugal treating：By the high speed tubular type centrifuge of extract solution 20000rpm/min under (1) item, obtain
To supernatant；
(3) isolate and purify：By HPD45A type large pore resin absorption columns on the supernatant under (2) item, first with 10 times of post beds
The mass concentration of volume is 5% ethanol elution, removes non-target components；It is again 16% with the mass concentration of 40 times of bed volumes
Ethanol elution, obtain active component eluent；
(4) concentrate：The active component under (3) item is concentrated under reduced pressure to give active component concentrate；
(5) dry：The active component concentrate under (4) item is dried with spray dryer, produces the effective portion of Swertia mussotii
Position.Tibetan capillary extract manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention, it is as a result as follows：Gentiamarin,
The content of 4 main compounds of sweroside mangiferin and Swertiamarin accounts for the 70.5% of active component solid content, four
The mass ratio of kind composition is 66:1.0:0.5:3.
Embodiment 9
It is prepared by the tablet of Swertia mussotii active component
Prescription：
Preparation method：The Swertia mussotii active component of Example 5, add cornstarch, lactose, using 95% ethanol as glue
Mixture, cross 20 mesh sieves and softwood is made, 60 DEG C of dryings, whole grain, after stiffened fatty acid magnesium mixes, be pressed into 2000, produce.Every contains
There is a Swertia mussotii active component 50mg, every equivalent to containing gentiamarin 40mg.
The pharmaceutical composition of Swertia mussotii active component manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention,
As a result it is as follows：Gentiamarin, sweroside, the mass ratio of mangiferin and Swertiamarin are 80:10:3.2:1.8.
Embodiment 10
It is prepared by the capsule of Swertia mussotii active component
Prescription：
Preparation method：The Swertia mussotii active component of Example 5, cornstarch is added, 95% ethanol is as adhesive, mistake
Softwood is made in 20 mesh sieves, 60 DEG C of dryings, whole grain, after stiffened fatty acid magnesium mixes, is distributed into 2000, produces.Every contains Tibetan mattress
Old active component 50mg, every equivalent to containing gentiamarin 40mg.
The pharmaceutical composition of Swertia mussotii active component manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention,
As a result it is as follows：Gentiamarin, sweroside, the mass ratio of mangiferin and Swertiamarin are 80:10:3.2:1.8.
Embodiment 11
It is prepared by the granule of Swertia mussotii active component
Prescription：
Preparation method：The Swertia mussotii active component of Example 5, cornstarch and lactose are added, 95% ethanol is as viscous
Mixture, cross 20 mesh sieves and softwood is made, 60 DEG C of dryings, whole grain, after stiffened fatty acid magnesium mixes, be distributed into 500 bags, produce.Every bag contains
There is a Swertia mussotii active component 0.2g, every bag equivalent to containing gentiamarin 0.16g.
The pharmaceutical composition of Swertia mussotii active component manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention,
As a result it is as follows：Gentiamarin, sweroside, the mass ratio of mangiferin and Swertiamarin are 80:10:3.2:1.8.
Embodiment 12
It is prepared by the injection of Swertia mussotii active component
Prescription：
Preparation method：Swertia mussotii active component 100g and 1g the sodium salicylate dissolving of Example 5, add water for injection to fit
Amount, stirring and dissolving, then inject water to 10L.Regulation pH value is 6.0-7.0, is extremely clarified with 0.22 μm of filtering with microporous membrane, with
Every 10mL is filled in 10mL ampoule bottles, and sealing, pressure sterilizing 20 minutes, are produced at 120 DEG C.Every has containing Swertia mussotii
Position 100mg is imitated, every equivalent to containing gentiamarin 80mg.
The pharmaceutical composition of Swertia mussotii active component manufactured in the present embodiment is determined using the foregoing HPLC methods of the present invention,
As a result it is as follows：Gentiamarin, sweroside, the mass ratio of mangiferin and Swertiamarin are 80:10:3.2:1.8.
Experimental example
A kind of pharmacodynamic study for the Swertia mussotii active component for treating acute, chronic hepatitis
1st, experiment material
SD rats 36, body weight (180-220g), SPF levels, it is purchased from Qinglongshan experimental animal feeding base.In experimentation
Rat ad lib and drinking-water；ANIT purchases in Aladdin Reagent Company, and positive control medicine Swertia mussotii piece is big in Qinghai Shandong
Ground pharmaceutcal corporation, Ltd；ALT, AST, TBil, DBil, γ-GT kits are purchased builds up biological Co., Ltd in Nanjing.
2nd, experimental design
After adaptability is fed three days, weigh, rat body weight 180-220g.Rat is divided into six groups, every group 6, specific point
Group information is as follows：
Blank group (Control)：Rat does not have to ANIT modelings and administration is intervened.
Model group (Model)：Rat uses ANIT gavage modelings, but does not receive pharmaceutical intervention.
Low dosage administration group (High)：After rat uses ANIT gavage modelings, receive 0.04g/kg medicine gavage to be measured
Treatment.
Middle dosage administration group (Medium)：After rat uses ANIT gavage modelings, the medicine to be measured for receiving 0.16g/kg fills
Stomach is treated.
High dose group (High)：After rat uses ANIT gavage modelings, receive 0.64g/kg medicine gavage to be measured treatment.
Positive drug group (positive)：After rat uses ANIT gavage modelings, the positive control medicine for receiving 0.8g/kg is hidden
Oriental wormwood piece gavage is treated.
ANIT is used using the preceding ANIT solution that 10g/L is made into sesame oil for induced rat acute, chronic hepatitis model.It is real
In testing, in addition to normal group gives the control of sesame oil gavage, remaining each group rat is respectively with 10mL/kg dosage in administration the previous day
With administration induction modeling in the 5th day.Starting gastric infusion after administration group and positive controls modeling one day, (model control group is given
The physiological saline of same dose).Continuous one week.After last dose, Rat Fast 12h.Anesthetized rat, eyeground vein clump take blood,
30min is stood at room temperature, 3000rpm/min centrifugation 30min, separates serum, -20 DEG C of preservations.For hepato-biliary function biochemical indicator
TBIL,DBIL,ALT,AST,γ-GT.Abdominal cavity is opened, separates the liver of rat, residual blood is cleaned, weighs.Take partial liver more than 4%
Polyformaldehyde is fixed, and is detected for pathological section.Remaining liver is in -80 DEG C of preservations.
3rd, the measure of experimental index
The body weight of rat and experiment terminate rear liver weight and the record of liver index in experimentation.
The detection of Biochemical Indices In Serum：ALT, AST, TBil, DBil, γ-GT are measured according to the specification of kit.
Hepatic histopathology is observed：Rats'liver reuses 4% paraformaldehyde and fixed, draw materials, be dehydrated, waxdip, embedding, then be cut into
4 μ m-thicks are cut into slices, and H＆E dyeing, optical microphotograph Microscopic observation is simultaneously taken pictures.
4th, data analysis
Experimental data is usedRepresent.Statistical analysis and icon are complete with Graph Pad Prismsoftware
Into.
5th, result and its analysis
(1) body weight of rat with administration time variation tendency
The body weight of rat with the variation tendency of administration time as shown in figure 3, as seen from the figure, at the 3rd day middle dose group with it is empty
There is conspicuousness change in white group rat body weight；Start model group and administration group within 4th day and conspicuousness occur with blank group rat
Change, wherein most obvious with middle dosage and high dose group.As a result show that ANIT can cause rat body when inducing acute, chronic hepatitis
Weight significantly reduces.
(2) the morphologic change of rat in experimentation
After last dose, take pictures, record rat morphological change.The morphologic change of rat is such as Fig. 4 institutes in experimentation
Show.Compared to rats in normal control group, model and the appearance of administration group rat are obvious One's spirits are drooping, and urine turns yellow, and hair is fluffy, abdomen
Portion's swelling, feed reduces and Body weight loss.And the abdomen swelling of administration group is improved, wherein the most obvious with low dose group.
(3) rat liver changes
After experiment terminates, rat liver is dissected, as shown in Figure 5.It was found that rats in normal control group liver surface is smooth,
Ruddy softness；Model group liver is congested, color and luster is dark yellow, and swelling is obvious.Compared with model group rats, administration group rat liver color and luster
More ruddy, congested swelling degree has mitigated.
(4) liver weight of each group rat and its corresponding liver index
After experiment terminates, rat liver is dissected, the liver weight of each group rat and its corresponding liver index are as shown in Figure 6.
It was found that rats in normal control group liver surface is smooth, ruddy softness；Model group liver is congested, color and luster is dark yellow, and swelling is obvious.
Compared with model group rats, administration group rat liver color and luster is more ruddy, and congested swelling degree has mitigated.Compared with blank group,
ANIT can notable induced rat body weight reduction, administration group, which goes out middle dosage, to be caused outside the faint reduction of liver weight, and other are given
There is the increase of liver weight in medicine group, and the increased conspicuousness of liver weight occurs in high dose group.Liver index is compared to blank
Group dramatically increases, and other each administration groups dramatically increase compared to the liver index of model group in addition to middle dose group.
(5) change of drug-induced ANIT rat blood serums biochemical indicator
The change of drug-induced ANIT rat blood serums biochemical indicator is as shown in Figure 7.As a result show, ANIT can be damaged significantly
The hepato-biliary function of rat, show as model group rats serum alt, AST content dramatically increase (hepatic injury) and TB, DB, γ-
GT content dramatically increases (choleresis function damage).The result of administration group shows that it is big that medicine to be measured can significantly improve ANIT
The hepatic injury of mouse, and its therapeutic effect is better than notable positive drug group (wherein the most obvious with low dose group).Medicine is to bile point
The improvement result of function is secreted, the improvement result of medicine and positive drug to be measured approaches, and the work of low dosage administration group and positive drug
With closest.Show based on the above results, administration group can significantly improve ANIT induced rat acute, chronic hepatitis, and medicine to be measured
Thing has more preferable improvement result compared with positive control drug.
(6) influence for the acute, chronic hepatitis liver tissues of rats morphological change that medicine is induced ANIT
The influence for the acute, chronic hepatitis liver tissues of rats morphological change that medicine is induced ANIT is as shown in Figure 8.Rats'liver
The H＆E coloration results explanation of dirty section, the liver organization form of normal rat is normal, no pathologic change.The mould of ANIT inductions
Type group rat liver tissue shows serious inflammatory cell infiltration, oedema, sinus extravasated blood, the necrosis of liver cells and leaflet of large area
Catheter injury.Administration group can significantly improve the lesion degree of hepatic tissue, wherein with the improvement result of low dosage medicine group to be measured most
To be obvious, and it is better than positive controls.
6th, conclusion
A kind of Swertia mussotii active component for treating acute, chronic hepatitis that the present invention obtains can significantly improve the liver of ANIT rats
Damage, and its therapeutic effect is significantly stronger than positive drug group, the H＆E coloration results explanation of rat liver section, the liver of normal rat
Dirty tissue morphology is normal, no pathologic change.
When description above illustrates of the invention, while the purpose for providing embodiment is to illustrate the reality of the present invention
Operating process and meaning of the present invention.When entering in the claims in the present invention and its equivalency range, reality of the invention should
With including all general changes, cooperation, or improvement.
Claims (9)
1. a kind of Swertia mussotii active component for treating acute, chronic hepatitis, it is characterised in that contain in the Swertia mussotii active component
Gentiamarin (Gentiopicroside), sweroside (Sweroside), mangiferin (Mangiferin) and Swertiamarin
(Swertiamrin) four kinds of active components, total content are 50~95%, wherein, the mass ratio of four kinds of active components is (40
~90)：(1.0~10)：(0.5~5)：(0.5~5).
A kind of 2. Swertia mussotii active component for treating acute, chronic hepatitis according to claim 1, it is characterised in that the dragon
Courage hardship glycosides, sweroside, the mass ratio of four kinds of active components of mangiferin and Swertiamarin are 80:10:3.2:1.8.
3. a kind of preparation method of Swertia mussotii active component for treating acute, chronic hepatitis according to claim 1, its feature
It is, comprises the following steps：
(1) extract：By Swertia mussotii pulverizing medicinal materials, with 1g：The ethanol that 5mL~30mL ratio mass concentration is 0~95% exists
Circumfluence distillation is carried out at 50~90 DEG C, extraction time is 1~3 time, and 1~3h, merging every time obtains extract solution；
(2) centrifugal treating：By 10000~20000rpm of extract solution high speed tubular type centrifuge, supernatant is obtained；
(3) purifies and separates：Large pore resin absorption column on supernatant will be obtained, is first 3 with the mass concentration of 5~20 times of bed volumes
~8% ethanol elution, remove non-target components；The ethanol for being again 10~20% with the mass concentration of 5~50 times of bed volumes
Elution, obtains active component eluent；
(4) concentrate：Above-mentioned active component is concentrated under reduced pressure or cut under the conditions of 0.05~0.1Mpa of vacuum, 50~90 DEG C of temperature
The organic film of the resistance to alcohol film that molecular weight is 100~1000 is stayed to concentrate；
(5) dry：Above-mentioned active component concentrate is dried, dried object, which crushes, produces Swertia mussotii active component.
4. a kind of preparation method of Swertia mussotii active component for treating acute, chronic hepatitis according to claim 3, its feature
It is, Swertia mussotii medicinal material described in step 1 is mussot swertia herb (Swertia mussotii), Swertia franchetiana H.Smith (Swertia
Chirayita), Indian Herba Swertiae bimaculatae (Swertia franchetiana), Swertia punicea Hemsl (Swertia punicea), ellipse
One or more of mixing in leaf flower anchor (Halenia elliptica), Marsh Felwort (Lomatogonium rotatum).
5. a kind of preparation method of Swertia mussotii active component for treating acute, chronic hepatitis according to claim 3, its feature
It is, the large pore resin absorption column used described in step 3 is AB-8, D101, S-8, X-5, D3520, HPD100, HPD300
With any one in HPD450A.
6. a kind of preparation method of Swertia mussotii active component for treating acute, chronic hepatitis according to claim 3, its feature
Be, described in step 5 dry for be dried under reduced pressure or microwave drying or freeze-drying or spray drying.
7. a kind of pharmaceutical composition for treating acute, chronic hepatitis, it is characterised in that described in the claim 1 comprising therapeutically effective amount
A kind of Swertia mussotii active component, pharmaceutic adjuvant and pharmaceutical carrier for treating acute, chronic hepatitis.
A kind of 8. described pharmaceutical composition for treating acute, chronic hepatitis according to right 7, it is characterised in that the medicine
Composition can be made into clinically acceptable tablet, capsule, injection, granule, pill, supensoid agent, dripping pill, liquid oral
Any one in preparation, aerosol, suppository or subcutaneous administration formulation.
9. a kind of Swertia mussotii active component for treating acute, chronic hepatitis according to claim 1 is preparing treatment acute and chronic liver
Application in scorching medicine, hepatic injury, removing jaundice can be significantly improved and reduce transaminase effect.
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| CN202210724960.7A CN115120605A (en) | 2017-11-07 | 2017-11-07 | Tibetan capillary artemisia effective part for treating acute and chronic hepatitis and preparation method and application thereof |
| CN201711081438.7A CN107875163A (en) | 2017-11-07 | 2017-11-07 | A kind of Swertia mussotii active component for treating acute, chronic hepatitis and its preparation method and application |
| AU2018101643A AU2018101643A4 (en) | 2017-11-07 | 2018-11-02 | Effective part of tibetan capillaris for treating acute and chronic hepatitis and its preparation method and application |
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| CN109793812A (en) * | 2018-12-19 | 2019-05-24 | 内蒙古民族大学 | Rib column flower extract and preparation method thereof, pharmaceutical composition and weight-reducing purposes |
| CN110407894A (en) * | 2019-07-12 | 2019-11-05 | 楚雄医药高等专科学校 | The extracting method of effective component in a kind of two capillus of Yi nationality's medicine |
| CN111514185A (en) * | 2020-06-12 | 2020-08-11 | 西藏天虹科技股份有限责任公司 | Tibetan capillary artemisia particle and preparation method thereof |
| CN112237585A (en) * | 2020-09-08 | 2021-01-19 | 中国科学院西北高原生物研究所 | Effective component of total picroside of Tibetan capillary artemisia for treating chronic cholecystitis and preparation method and application thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109793812A (en) * | 2018-12-19 | 2019-05-24 | 内蒙古民族大学 | Rib column flower extract and preparation method thereof, pharmaceutical composition and weight-reducing purposes |
| CN110407894A (en) * | 2019-07-12 | 2019-11-05 | 楚雄医药高等专科学校 | The extracting method of effective component in a kind of two capillus of Yi nationality's medicine |
| CN111514185A (en) * | 2020-06-12 | 2020-08-11 | 西藏天虹科技股份有限责任公司 | Tibetan capillary artemisia particle and preparation method thereof |
| CN111514185B (en) * | 2020-06-12 | 2021-12-07 | 西藏天虹科技股份有限责任公司 | Tibetan capillary artemisia particle and preparation method thereof |
| CN112237585A (en) * | 2020-09-08 | 2021-01-19 | 中国科学院西北高原生物研究所 | Effective component of total picroside of Tibetan capillary artemisia for treating chronic cholecystitis and preparation method and application thereof |
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| CN115120605A (en) | 2022-09-30 |
| AU2018101643A4 (en) | 2018-12-06 |
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Application publication date: 20180406 |