CN107737155A - A kind of Chinese mesona herb extract and its preparation method and application - Google Patents
A kind of Chinese mesona herb extract and its preparation method and application Download PDFInfo
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- CN107737155A CN107737155A CN201711224662.7A CN201711224662A CN107737155A CN 107737155 A CN107737155 A CN 107737155A CN 201711224662 A CN201711224662 A CN 201711224662A CN 107737155 A CN107737155 A CN 107737155A
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- jelly
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- 239000000284 extract Substances 0.000 title claims abstract description 262
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 241001646834 Mesona Species 0.000 title description 199
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 552
- 235000015110 jellies Nutrition 0.000 claims abstract description 117
- 239000008274 jelly Substances 0.000 claims abstract description 117
- 238000010828 elution Methods 0.000 claims abstract description 113
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 91
- 238000000605 extraction Methods 0.000 claims abstract description 34
- 238000010992 reflux Methods 0.000 claims abstract description 25
- 238000004440 column chromatography Methods 0.000 claims abstract description 10
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- 239000003125 aqueous solvent Substances 0.000 claims abstract description 6
- 229940077274 Alpha glucosidase inhibitor Drugs 0.000 claims abstract description 3
- 239000003888 alpha glucosidase inhibitor Substances 0.000 claims abstract description 3
- 239000003480 eluent Substances 0.000 claims abstract description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 44
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 42
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 23
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- XUGPCRRUMVWELT-UHFFFAOYSA-N 2-(2,5-dimethoxy-4-propan-2-ylphenyl)ethanamine Chemical compound COC1=CC(C(C)C)=C(OC)C=C1CCN XUGPCRRUMVWELT-UHFFFAOYSA-N 0.000 claims 1
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- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- IFBHRQDFSNCLOZ-IIRVCBMXSA-N 4-nitrophenyl-α-d-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC1=CC=C([N+]([O-])=O)C=C1 IFBHRQDFSNCLOZ-IIRVCBMXSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
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- 229920002527 Glycogen Polymers 0.000 description 1
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- 102400000471 Isomaltase Human genes 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
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- 108010026867 Oligo-1,6-Glucosidase Proteins 0.000 description 1
- NEZJDVYDSZTRFS-RMPHRYRLSA-N Phenyl beta-D-glucopyranoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1 NEZJDVYDSZTRFS-RMPHRYRLSA-N 0.000 description 1
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- 231100000053 low toxicity Toxicity 0.000 description 1
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- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
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- 201000008383 nephritis Diseases 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 239000003182 parenteral nutrition solution Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
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- 229920001282 polysaccharide Polymers 0.000 description 1
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- 102000004169 proteins and genes Human genes 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- 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)
- A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明属于医药领域，公开了一种凉粉草提取物及其制备方法和应用。本发明所述凉粉草提取物为凉粉草水提取物或凉粉草乙醇提取物。本发明所述凉粉草提取物的制备方法为取凉粉草的干燥全草粉碎，加水性溶剂回流提取，去除不溶物后干燥得到凉粉草初提物，凉粉草初提物进行大孔树脂开放柱层析，洗脱液洗脱。所述制备方法操作简单，易于实现，原料为天然产物，毒副作用小，制得的α‑葡萄糖苷酶活性抑制组分含量较大且抗α‑葡萄糖苷酶作用强，具有很好的α‑葡萄糖苷酶抑制作用，可作为α‑葡萄糖苷酶抑制剂用于治疗由α‑葡萄糖苷酶功能紊乱造成的一系列疾病。The invention belongs to the field of medicine, and discloses a jelly jelly extract, a preparation method and application thereof. The jelly jelly extract of the present invention is jelly jelly water extract or jelly jelly alcohol extract. The preparation method of the jelly jelly extract of the present invention is to take the dried whole herb of jelly jelly, pulverize it, add an aqueous solvent to reflux extraction, remove insoluble matter and then dry to obtain the first extract of jelly jelly, and the first extract of jelly jelly is subjected to a macroporous resin open column Chromatography, eluent elution. The preparation method is simple to operate and easy to implement, the raw materials are natural products, and the toxic and side effects are small, and the prepared α-glucosidase activity inhibitory component has a large content and strong anti-α-glucosidase effect, and has good α-glucosidase activity. Glucosidase inhibition, can be used as an α-glucosidase inhibitor for the treatment of a series of diseases caused by α-glucosidase dysfunction.
Description
Technical field
The invention belongs to field of medicaments, and in particular to a kind of Chinese mesona herb extract and its preparation method and application, especially
Application in as alpha-glucosidase restrainer.
Background technology
Alpha-glucosidase is one kind of hydrolase, is the general name of class of enzymes, and it can be from containing phlorose glycosidic bond bottom
Non-reducing end catalytically hydrolyzing alpha-Isosorbide-5-Nitrae-glucoside bond of thing, discharges glucose, or the glucose residue that of dissociating shifts
α -1,6- glucoside bonds, so as to obtain the phlorose such as oligoisomaltose or sugar ester, glycopeptide are formed to another saccharide substrates
Glycosidic bond enzyme.Alpha-glucosidase is widely distributed in nature, and species is various, distinct, is almost present in all organisms,
The glycogen degradation of the mankind and animal, plant, microorganism carbohydate metabolism in terms of there is important physiological function.Common α-Portugal
Polyglycoside enzyme includes maltose, isomaltase, invertase and trehalase etc..Alpha-glucosidase is deposited extensively in living nature
And participate in the structure and composition of glycometabolism, sugar chain, be the recognition site of protein exceptional function, the activity of glycosidase is to sugared egg
White biosynthesis has key effect, and it is not only the key enzyme of life entity normal operation, while is the phase of many diseases again
Enzyme is closed, it is relevant with a series of metabolic disturbance diseases.Therefore research alpha-glucosidase pair determines the structure of complicated polysaccharide
And related bioprocess is significant.
The treatment for carrying out disease by suppressing the method for enzyme activity at present has become a focus in drug research field.Enzyme
Inhibitor is important regulatory factor in vivo, and people develop the element task of zymoprotein, to the glucosides of high selectivity
The research of enzyme inhibitor can be not only used for studying the structure of enzyme active center, the mechanism of action of enzyme, the producing cause of disease, together
When them is expected to the medicine as many diseases.Suitable inhibitor is found to be used to controlling and adjusting alpha-glucosaccharase
A series of activity of enzyme, so as to which metabolic disturbance diseases are carried out with research heat of the treatment as biology and medical domain
Point.
The screening and development of alpha-glucosidase restrainer, which start from state, its source such as Germany and Japan, can be divided into three major types：
The inhibitor of natural plant extracts, microorganism extracts and artificial chemistry synthesis.At present, split alpha-glucosidase restrainer
The study hotspot of class medicine is gradually turned to the natural products of high-efficiency low-toxicity by artificial synthesized compound.China's traditional Chinese medicine resource is rich
Richness, a huge medical treasure-house is provided to develop alpha-glucosidase inhibitor medicine.
Chinese mesona herb (Mesona chinensis Benth.), also known as Mesona chinensis Benth, Chinese mesona herb, celestial being's jelly, are Labiatae
(Labiatae) Chinese mesona herb category (MesonaBl.) plant, it is a kind of east plant resources of important medicine-food two-purpose, main distribution
In China Guangdong, Guangxi, Jiangxi, Fujian, Zhejiang, Taiwan and Yunnan and other places.According to ancient medicine book《Book on Chinese herbal medicine asks former》Record, bean jelly
Grass has the effect of " clear hot summer weather, Xie Zangfu, tie heat toxin, control wine wind ",《Gather medicinal herbs in the south of the Five Ridges》Described in, Chinese mesona herb can control colored willow poison and enter
Bone.《Dictionary of medicinal plant》Record：Chinese mesona herb nature and flavor are puckery, sweet, cold, have clear heat quench one's thirst, cool blood relieving summer-heat and the effect of diuresis, it is among the people often
As hot summer days cold drink, and available for acute wind-warm syndrome arthritis, hypertension, heatstroke, diabetes, acute nephritis, flu, Huang
Subcutaneous ulcer, scald etc. are treated.Modern pharmacology research shows that the extract of Chinese mesona herb has the effect such as anti-oxidant, antibacterial.
The content of the invention
It is an object of the present invention to provide a kind of Chinese mesona herb extract.
Chinese mesona herb extract of the present invention is Chinese mesona herb water extract and/or Chinese mesona herb ethanol extract.
Wherein, the Chinese mesona herb water extract is Chinese mesona herb water primary extract, the ethanol elution group of Chinese mesona herb water primary extract 0%
Divide, the ethanol elution component of Chinese mesona herb water primary extract 20%, the ethanol elution component of Chinese mesona herb water primary extract 40%, Chinese mesona herb water just carry
The ethanol elution component of thing 60%, the ethanol elution component of Chinese mesona herb water primary extract 80%, the ethanol elution of Chinese mesona herb water primary extract 100%
At least one of component.
The Chinese mesona herb ethanol extract is the ethanol primary extract of Chinese mesona herb 60%, the second of 60% ethanol primary extract of Chinese mesona herb 0%
Alcohol elution fraction, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 10%, the ethanol of 60% ethanol primary extract of Chinese mesona herb 30%
Elution fraction, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 50%, the ethanol of 60% ethanol primary extract of Chinese mesona herb 70% are washed
De- component, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 95%, the ethanol primary extract of Chinese mesona herb 80%, Chinese mesona herb chloroform layer
At least one in extract, Chinese mesona herb ethyl acetate layer extract, Chinese mesona herb n-butanol layer extract, Chinese mesona herb water layer extract
Kind.
The second object of the present invention is the preparation method for providing the Chinese mesona herb extract.
A kind of preparation method of Chinese mesona herb extract, take the drying herb of Chinese mesona herb to crush, add aqueous solvent refluxing extraction,
Chinese mesona herb primary extract is dried to obtain after removing insoluble matter, Chinese mesona herb primary extract carries out macroreticular resin and opens column chromatography, and eluent is washed
It is de-.
Wherein, preferably, the aqueous solvent is water or ethanol water.
Preferably, the removal insoluble matter is specially collected by filtration filtrate or supernatant is collected by centrifugation.
In some embodiments, the preparation method of the Chinese mesona herb extract is specially the drying herb powder of Chinese mesona herb
It is broken, add water refluxing extraction, filter, filtrate is dried to obtain Chinese mesona herb water primary extract, and Chinese mesona herb water primary extract carries out macroreticular resin and opened
Column chromatography is put, carries out gradient with the ethanol water that percent by volume is 0%, 20%, 40%, 60%, 80% and 100% successively
Elution, obtains Chinese mesona herb water extract.Obtain the ethanol elution component of Chinese mesona herb water primary extract 0%, Chinese mesona herb water primary extract
20% ethanol elution component, the ethanol elution component of Chinese mesona herb water primary extract 40%, the ethanol elution group of Chinese mesona herb water primary extract 60%
Point, the ethanol elution component of Chinese mesona herb water primary extract 80% and the ethanol elution component of Chinese mesona herb water primary extract 100%.
In some embodiments, the preparation method of the Chinese mesona herb extract is specially the drying herb powder of Chinese mesona herb
It is broken, the extraction of 60% alcohol reflux, filtering are added, filtrate is dried to obtain the ethanol primary extract of Chinese mesona herb 60%, the ethanol of Chinese mesona herb 60%
Primary extract carries out macroreticular resin and opens column chromatography, is successively 0%, 10%, 30%, 50%, 70% and 95% with percent by volume
Ethanol water carry out gradient elution, obtain the ethanol extract of Chinese mesona herb 60%.Obtain the ethanol primary extract of Chinese mesona herb 60%
0% ethanol elution component, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 10%, the ethanol primary extract 30% of Chinese mesona herb 60%
Ethanol elution component, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 50%, the second of 60% ethanol primary extract of Chinese mesona herb 70%
Alcohol elution fraction and the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 95%.
In some embodiments, the preparation method of the Chinese mesona herb extract is specially the drying herb powder of Chinese mesona herb
It is broken, the extraction of 80% alcohol reflux is added, filtering, filtrate is dried to obtain the ethanol primary extract of Chinese mesona herb 80%, by the second of Chinese mesona herb 80%
Alcohol primary extract is distributed in water into the aqueous solution, is extracted, is concentrated to give with isometric chloroform, ethyl acetate and water-saturated n-butanol
Chinese mesona herb chloroform layer extract, Chinese mesona herb ethyl acetate layer extract, Chinese mesona herb n-butanol layer extract, the extraction of Chinese mesona herb water layer
Thing.
Preferably, the refluxing extraction is heat extraction or cold extraction.It will be understood by those skilled in the art that the backflow
Extraction includes but is not limited to the methods of cold soaking extraction, seepage extraction, surname extraction and circumfluence distillation.
Preferably, the addition of the aqueous solvent is 8~15 times of Chinese mesona herb weight, Extracting temperature is 10~100
DEG C, extraction time is 2~6 times, and each extraction time is 1~72 hour.
In some embodiments, described plus water refluxing extraction is to add the water of 10 times of Chinese mesona herb weight, refluxing extraction three
It is secondary, 2 hours every time, merge extract solution.
In some embodiments, 60% second for adding 60% alcohol reflux and being extracted as adding 12 times of Chinese mesona herb weight
Alcohol reflux extracts twice, 2 hours every time, merges extract solution.
In some embodiments, 80% second for adding 80% alcohol reflux and being extracted as adding 12 times of Chinese mesona herb weight
Alcohol reflux extracts twice, 2 hours every time, merges extract solution；
Preferably, the drying is to be dried under reduced pressure.
Preferably, the extraction extracts three times respectively for chloroform, ethyl acetate and water-saturated n-butanol.
Further, the step of concentration is also included after the extraction.Preferably, the concentration is to be concentrated under reduced pressure.
In some embodiments, the present invention have detected the Chinese mesona herb extract and alpha-glucosaccharase enzyme level lived respectively
As a result property shows that the Chinese mesona herb extract is inhibited to alpha-glucosidase to alpha-glucosaccharase enzyme inhibition activity,
And inhibition is better than positive control (acarbose), therefore present invention also offers Chinese mesona herb extract as phlorose
Application in glycosides enzyme inhibitor.
Preferably, the Chinese mesona herb extract as alpha-glucosidase restrainer is Chinese mesona herb water extract or cool
Powder grass ethanol extract.
Wherein, the Chinese mesona herb water extract as alpha-glucosidase restrainer is preferably Chinese mesona herb water primary extract
20% ethanol elution component, the ethanol elution component of Chinese mesona herb water primary extract 40%, the ethanol elution group of Chinese mesona herb water primary extract 60%
Point, at least one in the ethanol elution component of Chinese mesona herb water primary extract 80%, the ethanol elution component of Chinese mesona herb water primary extract 100%
Kind.
The Chinese mesona herb ethanol extract as alpha-glucosidase restrainer is the ethanol primary extract of Chinese mesona herb 60%, cool
Powder grass 60% ethanol primary extract 10% ethanol elution component, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 30%, bean jelly
The careless ethanol elution component of 60% ethanol primary extract 50%, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 70%, Chinese mesona herb
The ethanol elution component of 60% ethanol primary extract 95%, the ethanol primary extract of Chinese mesona herb 80%, Chinese mesona herb chloroform layer extract, Chinese mesona herb
At least one of ethyl acetate layer extract, Chinese mesona herb n-butanol layer extract, Chinese mesona herb water layer extract.
Further, present invention also offers above-mentioned the Chinese mesona herb extract and medicine of a kind of pharmaceutical preparation, including effective dose
Acceptable auxiliary material on.
Preferably, the pharmaceutical preparation can pharmaceutically be connect by the above-mentioned Chinese mesona herb extracts of 10%-90% and 10%-90%
The auxiliary material composition received.
Those skilled in the art can prepare the Chinese mesona herb extract direct or indirect addition required during different dosage forms
Pharmaceutically acceptable various conventional auxiliary materials, such as filler, disintegrant, lubricant, adhesive, with traditional drug formulations side
Method, conventional oral formulations or ejection preparation are made.
Preferably, the oral formulations are tablet, capsule, granule.
Preferably, the ejection preparation is parenteral solution or powder-injection.
In some embodiments, the pharmaceutical preparation is the ethanol primary extract granule of Chinese mesona herb 60%, by following weight
Part raw material is made：200 parts of 60% ethanol primary extract of Chinese mesona herb, 150 parts of lactose, 30 parts of 10% starch slurry.
In some embodiments, the pharmaceutical preparation is the ethanol elution component glue of 60% ethanol primary extract of Chinese mesona herb 95%
Wafer, it is made up of following raw material：80 parts of 60% ethanol primary extract of Chinese mesona herb, 95% ethanol elution component, 60 parts of lactose,
60 parts of microcrystalline cellulose, 4 parts of talcum powder.
In some embodiments, the pharmaceutical preparation is Chinese mesona herb chloroform layer extract tablet, former by following parts by weight
Material is made：150 parts of Chinese mesona herb chloroform layer extract, 120 parts of lactose, 16 parts of 10% starch slurry, 40 parts of PVPP, stearic acid
4 parts of magnesium.
As shown from the above technical solution, the invention provides a kind of Chinese mesona herb extract and its preparation method and application.This
It is Chinese mesona herb water extract or Chinese mesona herb ethanol extract to invent the Chinese mesona herb extract.Chinese mesona herb extract of the present invention
Preparation method be that the drying herb for taking Chinese mesona herb crushes, add aqueous solvent refluxing extraction, remove be dried to obtain after insoluble matter it is cool
Powder grass primary extract, Chinese mesona herb primary extract carry out macroreticular resin and open column chromatography, elution.The preparation method operation letter
It is single, it is easy to accomplish, raw material is natural products, and toxic side effect is small, and obtained alpha-glucosidase activity constituents for suppressing content is larger
And anti-alpha-glucosidase effect is by force, has good alpha-glucosaccharase enzyme inhibition, can be used as alpha-glucosaccharase enzyme level
Agent is used to treat a series of diseases as caused by alpha-glucosidase dysfunction.
Embodiment
The invention discloses a kind of Chinese mesona herb extract and its preparation method and application.Those skilled in the art can use for reference
Present disclosure, it is suitably modified technological parameter realization.In particular, all similar replacements and change are to this area skill
It is it will be apparent that they are considered as being included in the present invention for art personnel.The present invention method and product by compared with
Good embodiment is described, related personnel substantially can not depart from present invention, in spirit and scope to as described herein
Method is modified or suitably changed with combining, to realize and using the technology of the present invention.
For a further understanding of the present invention, below in conjunction with the embodiment of the present invention, to the technical side in the embodiment of the present invention
Case is clearly and completely described, it is clear that and described embodiment is only part of the embodiment of the present invention, rather than all
Embodiment.Based on the embodiment in the present invention, those of ordinary skill in the art institute under the premise of creative work is not made
The every other embodiment obtained, belongs to the scope of protection of the invention.
Unless otherwise specified, reagent involved in the embodiment of the present invention is commercially available prod, can pass through business canal
Road purchase obtains.
The preparation of embodiment 1, Chinese mesona herb water primary extract and each separation component
(1) by 2 kilograms of the drying herb crushing of Chinese mesona herb, with the water refluxing extraction of 10 times of Chinese mesona herb weight three times, extraction
100 DEG C of temperature, 2 hours every time, merge extract solution；
(2) filter off except insoluble matter, be dried under reduced pressure, obtain Chinese mesona herb water primary extract 480g；
(3) Chinese mesona herb water primary extract 400g is subjected to macroreticular resin and opens column chromatography, be successively 0% with percent by volume,
20%th, 40%, 60%, 80% and 100% ethanol water carries out gradient elution, obtains the ethanol of Chinese mesona herb water primary extract 0%
Elution fraction 130g, the ethanol elution component 80g of Chinese mesona herb water primary extract 20%, the ethanol elution component of Chinese mesona herb water primary extract 40%
60g, the ethanol elution component 40g of Chinese mesona herb water primary extract 60%, the ethanol elution component 30g of Chinese mesona herb water primary extract 80% and bean jelly
The careless ethanol elution component 10g of water primary extract 100%.
The preparation of embodiment 2, the ethanol primary extract of Chinese mesona herb 60% and each separation component
(1) by 4 kilograms of crushing of drying herb of Chinese mesona herb, two are extracted with 60% alcohol reflux of 12 times of Chinese mesona herb weight
It is secondary, 80 DEG C of Extracting temperature, 2 hours every time, merge extract solution；
(2) filter off except insoluble matter, be dried under reduced pressure, obtain the ethanol primary extract 1050g of Chinese mesona herb 60%；
(3) the ethanol primary extract 1000g of Chinese mesona herb 60% is subjected to macroreticular resin and opens column chromatography, use percent by volume successively
Gradient elution is carried out for 0%, 10%, 30%, 50%, 70% and 95% ethanol water, at the beginning of obtaining the ethanol of Chinese mesona herb 60%
The ethanol elution component 210g of extract 0%, the ethanol elution component 150g of 60% ethanol primary extract of Chinese mesona herb 10%, the second of Chinese mesona herb 60%
The ethanol elution component 120g of alcohol primary extract 30%, the ethanol elution component 180g of 60% ethanol primary extract of Chinese mesona herb 50%, Chinese mesona herb
The ethanol elution component 125g of the 60% ethanol primary extract 70% and ethanol elution component 25g of 60% ethanol primary extract of Chinese mesona herb 95%.
The preparation of embodiment 3, the ethanol primary extract of Chinese mesona herb 80% and each separation component
(1) by 4.5 kilograms of crushing of drying herb of Chinese mesona herb, two are extracted with 80% alcohol reflux of 12 times of Chinese mesona herb weight
It is secondary, 70 DEG C of Extracting temperature, 2 hours every time, merge extract solution；
(2) filter off except insoluble matter, be dried under reduced pressure, obtain the ethanol primary extract 1230g of Chinese mesona herb 80%；
(3) the ethanol primary extract 1000g of Chinese mesona herb 80% is distributed in water into the 10L aqueous solution, with isometric chloroform,
Ethyl acetate and water-saturated n-butanol extract three times respectively, are concentrated under reduced pressure to give Chinese mesona herb chloroform layer extract 43g, Chinese mesona herb second
Ethyl acetate layer extract 52g, Chinese mesona herb n-butanol layer extract 65g, Chinese mesona herb water layer extract 700g.
Embodiment 4, Chinese mesona herb water primary extract and each separation component are to alpha-glucosaccharase enzyme inhibition activity
Chinese mesona herb water primary extract made from Example 1 and each separation component：Chinese mesona herb water primary extract, Chinese mesona herb water just carry
The ethanol elution component of thing 0%, the ethanol elution component of Chinese mesona herb water primary extract 20%, the ethanol elution group of Chinese mesona herb water primary extract 40%
Divide, at the beginning of the ethanol elution component of Chinese mesona herb water primary extract 60%, the ethanol elution component of Chinese mesona herb water primary extract 80% and Chinese mesona herb water
The ethanol elution component of extract 100%, it is as follows to alpha-glucosaccharase enzyme inhibition activity, specific testing procedure to test it.
(1) each sample prepares different concentration gradients respectively with biological level DMSO (dimethyl sulfoxide (DMSO)) respectively.
(2) toward adding the phosphate buffer solution (PBS) that 92 μ L concentration are 0.1mol/L, pH=6.8 in 96 hole microwell plates,
The sample solution of 10 μ L various concentrations is taken to add in each micropore respectively with liquid-transfering gun again, blank control group then adds the DMSO of equivalent
(dimethyl sulfoxide (DMSO)).After well mixed, the μ L of alpha-glucosaccharase enzyme solutions 8 that concentration is 0.1U/ml are added, do three groups of parallel realities
Test.And background group then directly adds 100 μ L concentration to be 0.1mol/L, pH=6.8 phosphate buffer solution (PBS), then take respectively
The sample of 10 μ L various concentrations is added in each micropore.Add and 96 microwell plates are transferred in multi-function microplate reader immediately, at 37 DEG C
Constant-temperature incubation 10min, wherein annular shaking 30s；
After the completion of (3) 37 DEG C of constant-temperature incubations, 96 microwell plates are taken out, add 20 μ L α-to nitro toward each hole with the volley of rifle fire immediately
Phenol glucoside (PNPG), is immediately placed into multi-function microplate reader afterwards, 37 DEG C of constant-temperature incubation 30min, wherein annular shaking
30s；
(4) after the completion of being incubated, 96 microwell plates are taken out, it is 1mol/L to add 70 μ L concentration toward each hole with the volley of rifle fire immediately after
Sodium carbonate liquor stops reaction, and microwell plate is put into multi-function microplate reader at once, annular shaking 30s；
(5) under 405nm wavelength, the OD values in each hole are determined, calculate the average value of three groups of repeat samples, and according to formula meter
Calculate under each sample various concentrations to alpha-glucosaccharase enzyme inhibition rate.
Note：ABlankFor the absorbance of blank control group；ASampleFor the absorbance of sample sets；ABackgroundFor the absorbance of background group.
(6) using inhibiting rate as Y-axis coordinate, sample concentration is X-axis coordinate, and being fitted log10 curves with origin softwares distinguishes
Suppression curve of each sample to alpha-glucosidase is drawn out, each sample is calculated to alpha-glucosidase by origin softwares
Half-inhibition concentration IC50Value.According to IC50Value and suppression curve judge each sample whether have inhibitory action and its relative to
The inhibition of positive control (acarbose).Test result is shown in Table 1.
The IC of the Chinese mesona herb water primary extract of table 1 and each separation component to alpha-glucosidase50Value
As a result show, in addition to Chinese mesona herb water primary extract and the ethanol elution component of Chinese mesona herb water primary extract 0%, Chinese mesona herb water
Primary extract 20%, 40%, 60%, 80% and 100% ethanol elution component compared to positive control acarbose for, be respectively provided with
Very strong alpha-glucosidase activity inhibition.
Embodiment 5, the ethanol primary extract of Chinese mesona herb 60% and each separation component are to alpha-glucosaccharase enzyme inhibition activity
The ethanol primary extract of Chinese mesona herb 60% and each separation component made from Example 2：The ethanol primary extract of Chinese mesona herb 60%,
The ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 0%, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 10%, bean jelly
The careless ethanol elution component of 60% ethanol primary extract 30%, the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 50%, Chinese mesona herb
The ethanol elution component of 60% ethanol primary extract 70% and the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 95%.It is right to test its
Alpha-glucosaccharase enzyme inhibition activity, for specific testing procedure with embodiment 4, test result is shown in Table 2.
The IC of the ethanol primary extract of 2 Chinese mesona herb of table 60% and each separation component to alpha-glucosidase50Value
As a result show, in addition to the ethanol elution component of 60% ethanol primary extract of Chinese mesona herb 0%, the ethanol of Chinese mesona herb 60% just carries
Thing and its 10%, 30%, 50%, 70% and 95% ethanol elution component compared to positive control acarbose for, be respectively provided with
Very strong alpha-glucosidase activity inhibition.
Embodiment 6, the ethanol primary extract of Chinese mesona herb 80% and each separation component are to alpha-glucosaccharase enzyme inhibition activity
The ethanol primary extract of Chinese mesona herb 80% and extract made from Example 3：The ethanol primary extract of Chinese mesona herb 80%, bean jelly
Careless chloroform layer extract, Chinese mesona herb ethyl acetate layer extract, Chinese mesona herb n-butanol layer extract, Chinese mesona herb water layer extract,
It is same to alpha-glucosaccharase enzyme inhibition activity, specific testing procedure to test it
Embodiment 4, test result is shown in Table 3.
The IC of the ethanol primary extract of 3 Chinese mesona herb of table 80% and each separation component to alpha-glucosidase50Value
As a result show, the ethanol primary extract of Chinese mesona herb 80%, Chinese mesona herb chloroform layer extract, the extraction of Chinese mesona herb ethyl acetate layer
Thing, Chinese mesona herb n-butanol layer extract, Chinese mesona herb water layer extract compared to positive control acarbose for, be respectively provided with very strong
Alpha-glucosidase activity inhibition.
Embodiment 7, the ethanol primary extract granule of Chinese mesona herb 60%
The ethanol primary extract 200mg of Chinese mesona herb 60%；
Lactose 150mg；
10% starch slurry 30mg；
The preparation process of the ethanol primary extract granule of Chinese mesona herb 60% is：
(1) the ethanol primary extract of Chinese mesona herb 60% first with lactose mixing 10-15 minutes；
(2) 10% starch slurry softwood is added, crosses 14 mesh sieves, is dried after granulation；
(3) 12 mesh sieves are crossed, is dried after whole grain, produces the ethanol primary extract granule of Chinese mesona herb water 60% of the present invention.
The ethanol elution component capsule of 8 Chinese mesona herb of embodiment, 60% ethanol primary extract 95%
The ethanol elution component 80mg of 60% ethanol primary extract of Chinese mesona herb 95%；
Lactose 60mg；
Microcrystalline cellulose 60mg；
Talcum powder 6mg；
The preparation process of the ethanol elution component capsule of Chinese mesona herb 95% is：
(1) the ethanol elution component of Chinese mesona herb 95% and lactose mixing 10-15 minutes；
(2) 10-15 minutes are mixed after adding microcrystalline cellulose；
(3) 3-5 minutes are mixed after adding talcum powder；
(4) mixture loads gelatine capsule shell, produces the ethanol elution component capsule of Chinese mesona herb 95% of the present invention.
The Chinese mesona herb chloroform layer extract tablet of embodiment 9
The preparation process of Chinese mesona herb chloroform layer extract tablet is：
(1) Chinese mesona herb chloroform layer extract and lactose mixing 10-15 minutes；
(2) 10% starch slurry softwood is added, crosses 14 mesh sieves, is dried after granulation, crosses 12 mesh sieve whole grains；
(3) and then PVPP and magnesium stearate are added, mixes 3-5 minutes；
(4) tabletting after being well mixed, produces Chinese mesona herb chloroform layer extract tablet of the present invention.
10 3 kinds of Chinese mesona herb extract formulations of embodiment are tested alpha-glucosaccharase enzyme inhibition activity
The ethanol of Chinese mesona herb 95% prepared by the ethanol primary extract granule of Chinese mesona herb 60% of the preparation of Example 7, embodiment 8
Chinese mesona herb chloroform layer extract tablet made from elution fraction Capsule content, embodiment 9, tests it to phlorose respectively
Glycosides enzyme inhibition activity, specific testing procedure are shown in Table 4 with the test result of embodiment 4.
IC of the 4 three kinds of Chinese mesona herb extract formulations of table to alpha-glucosidase50Value
As a result show, the ethanol primary extract granule of Chinese mesona herb 60%, the ethanol elution of 60% ethanol primary extract of Chinese mesona herb 95%
Component capsule and Chinese mesona herb chloroform layer extract tablet compared to positive control acarbose for, be respectively provided with very strong α-Portugal
Polyglycoside inhibition of enzyme activity effect.
It the foregoing is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, on the premise of raw material of the present invention is not departed from, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
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