CN108159195A - A kind of Leaf of Longestapex Beautyberry extract, preparation method and applications - Google Patents
A kind of Leaf of Longestapex Beautyberry extract, preparation method and applications Download PDFInfo
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- A61K2236/30—Extraction of the material
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- 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
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Abstract
The invention discloses a kind of Leaf of Longestapex Beautyberry extract, preparation method and applications, belong to technical field of natural product extraction.Mixture containing carnosol, rosmanol and different rosmanol in the Leaf of Longestapex Beautyberry extract.The invention also discloses the preparation method and applications of above-mentioned Leaf of Longestapex Beautyberry extract.In the Leaf of Longestapex Beautyberry extract of the present invention containing carnosol, rosmanol, different rosmanol mixture, these ingredients are diterpene phenols component, which has preferable anti-oxidant, anticancer and the various actives such as anti-inflammatory.By the pharmacodynamic study to Leaf of Longestapex Beautyberry extract, broader space will be provided for its further application.
Description
Technical field
The present invention relates to a kind of Leaf of Longestapex Beautyberry extract, preparation method and applications, belong to technical field of natural product extraction.
Background technology
Nitric oxide (nitric oxide, NO) is used as a kind of free radical, both the work(with second messenger and neurotransmitter
Can, and can be as the pathology and physiology course of a variety of diseases such as effector molecule transmitting inflammation, immune.In acute and chronic inflammation
In, NO is important pro-inflammatory cytokine.The experimental results show that the joint fluid of rheumatisant and serum NO are horizontal apparent high
In normal person.NO serves not only as the generation that a kind of inflammatory mediator directly takes part in rheumatism, and other inflammatory factors can be promoted to release
It puts, and then promotes articular cartilage damage, sb.'s illness took a turn for the worse for exacerbation.If reducing the generation of NO, protection can be provided for arthritis.
Leaf of Longestapex Beautyberry (Callicarpa longissima) be Verenaceae Japanses beauty-berry platymiscium, alias glue hand wind, wear bone maple,
Snow is prominent, year tongue it is wide etc..It is distributed in the ground such as Jiangxi, Fujian, Taiwan, Guangdong, Guangxi, Sichuan.Acrid flavour, slight bitter, it is warm-natured, enter
Lung, liver two pass through, and have the effect of hemostatic analgesia, eliminating stasis to subdue swelling, wind-damp dispelling, primary treatment traumatic hemorrhage, spitting of blood, cold after delivery pain, four
The illnesss such as acroparalysis, arthralgia pain due to rheumatism.
But it is at present, less to the research of Leaf of Longestapex Beautyberry both at home and abroad, more rarely have report to the application of Leaf of Longestapex Beautyberry active component.Cause
This, there is an urgent need for carrying out the research of Leaf of Longestapex Beautyberry active component extraction and purification process and its related application, to make up drawbacks described above.
Invention content
An object of the present invention is to provide a kind of Leaf of Longestapex Beautyberry extract.Contain mouse in the Leaf of Longestapex Beautyberry extract of the present invention
The mixture of tail grass phenol, rosmanol and different rosmanol, these ingredients are diterpene phenols component, which has preferable
Anti-oxidant, anticancer and the various actives such as anti-inflammatory.To be that it is further by the pharmacodynamic study to Leaf of Longestapex Beautyberry extract
Using the broader space of offer.
The technical solution that the present invention solves above-mentioned technical problem is as follows：A kind of Leaf of Longestapex Beautyberry extract, the Leaf of Longestapex Beautyberry extraction
Mixture containing three kinds of carnosol, rosmanol and different rosmanol in object, wherein, rat-tail in the Leaf of Longestapex Beautyberry extract
The mass content of careless phenol is 53.0-82.6%, and the mass percentage of rosmanol is 4.5- in the Leaf of Longestapex Beautyberry extract
12.4%, the mass percentage of different rosmanol is 2.1-8.0% in the Leaf of Longestapex Beautyberry extract
Carnosol, rosmanol and different rosmanol are diterpene phenols component.Pharmacological research shows in natural products
Diterpene phenols component there are the drug activities such as preferable anti-oxidant, antibacterial, anticancer, neuroprotection, anti-inflammatory, wherein in anti-inflammatory work
Property aspect, which can inhibit the formation of lipopolysaccharides, and targeting inhibits prostate in Microsomal prostaglandin E2 synthase
The synthesis of plain E2 synthase；Can also by inhibit Src/NF- κ B signals accesses and inflammatory factor IL-1 β and IL-6 expression and
Release plays anti-inflammatory activity.
Present inventor passes through the study found that being rich in carnosol, rosmanol and different fan in Leaf of Longestapex Beautyberry extract
The repeatedly diterpene phenols component such as fragrant phenol, and Pharmacological Results show that the extract has preferable anti-inflammatory activity, in addition, carnosol
It is a kind of compound with preferable anti-inflammatory activity, as the main component in Leaf of Longestapex Beautyberry extract, content is higher, can more play
Its anti-inflammatory activity.This discovery of the application will provide broader space for the further application of Leaf of Longestapex Beautyberry extract.
The second object of the present invention is to provide the preparation method of above-mentioned Leaf of Longestapex Beautyberry extract.The present invention is mixed using similar
Principle, the organic solvent of low polarity rapidly extracts diterpene phenols component from Leaf of Longestapex Beautyberry in utilization, and passes through column layer
Isolation technics is analysed, effectively removes depigmentation, is enriched with diterpene phenols component.
The technical solution that the present invention solves above-mentioned technical problem is as follows：A kind of preparation method of Leaf of Longestapex Beautyberry extract, including
Following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
Take dry Leaf of Longestapex Beautyberry powder, the organic solvent refluxing extraction of 10-20 times of addition Leaf of Longestapex Beautyberry powder quality 3 times, mistake
Filter, merging filtrate are concentrated under reduced pressure filtrate and obtain medicinal extract, as Leaf of Longestapex Beautyberry crude extract；
Step 2：Purifying
The Leaf of Longestapex Beautyberry crude extract that step 1 obtains by silica gel column chromatography is purified, grease-like composition is first removed with petroleum ether,
Again using volume ratio as (5-3):1 petroleum ether:The mixed solvent or volume ratio of ethyl acetate are (6-4):1 petroleum ether and acetone
Mixed solvent eluted, collect eluent, recycling design is dry to get to the Leaf of Longestapex Beautyberry extract of purifying；
Or the inverted column of material chromatographic purifying of Leaf of Longestapex Beautyberry crude extract for obtaining step 1, using volumetric concentration as 40-50%
Alcohols removal impurity, then alcohols using volumetric concentration as 75-95% elutes, and collects eluent, recycles solution, it is dry to get
To the Leaf of Longestapex Beautyberry extract of purifying.
Based on the above technical solution, the present invention can also be improved as follows.
Further, Leaf of Longestapex Beautyberry powder described in step 1 is after taking stem, branch, the leaf drying of raw material Leaf of Longestapex Beautyberry, to be crushed to 2mm institutes
.
It is using above-mentioned further advantageous effect：Suitable grain size not only contributes to the extraction of active ingredient, Er Qieyou
Conducive to the filtering of extracting solution.
Further, organic solvent described in step 1 is one kind in dichloromethane, chloroform, ethyl acetate, acetone.
It is using above-mentioned further advantageous effect：Above-mentioned organic solvent is middle low polar solvent, as Extraction solvent,
It can effective rapid extraction diterpene phenols component.
Further, the temperature extracted described in step 1 is 50-70 DEG C, every time 1 hour.
It is using above-mentioned further advantageous effect：Extracting temperature is milder, and active constituent is avoided to change because of high temperature.
Further, the aperture filtered described in step 1 is 30-70 μm.
It is using above-mentioned further advantageous effect：It being capable of effective fast filtering extracting solution.
Further, medicinal extract described in step 1 is 1.02-1.08 in 65 DEG C of proportion.
Further, the medium of reversed phase column chromatography described in step 2 is one kind in C18, C8, C4 or MCI.
Wherein, the grain size of above-mentioned C18 is 40-60 μm, and the grain size of C8 is 40-75 μm, and the grain size of C4 is 40-60 μm, MCI's
Grain size is 75-150 μm.
It is using above-mentioned further advantageous effect：The impurity such as the pigment in Leaf of Longestapex Beautyberry raw material, product color can be effectively removed
Pool is good.
Further, alcohols described in step 2 is methanol or ethyl alcohol.
It is using above-mentioned further advantageous effect：The methanol or ethanol solution of debita spissitudo can be effectively by reversed materials
In diterpene phenols component parse, diterpene phenols component is made more to concentrate.
The third object of the present invention is to provide the application of above-mentioned Leaf of Longestapex Beautyberry extract.The Leaf of Longestapex Beautyberry extract of the present invention can
To be applied to prepare anti-inflammatory drug or anti-inflammatory ancillary drug, there is wide application space.
The technical solution that the present invention solves above-mentioned technical problem is as follows：It is prepared by a kind of Leaf of Longestapex Beautyberry extract as described above
Application in anti-inflammatory drug or anti-inflammatory ancillary drug.
Dimethylbenzene inducing mouse ear swelling model is that certain inflammatory mediators such as histamine, kassinin kinin and plasmin is caused to explain
It puts, causes local vessel expansion, capillary permeability increases, and inflammatory cell infiltration causes ear's acute exudative inflammation oedema.
The model is widely used in evaluation plant extracts or synthesizes the anti-inflammatory activity effect of compound at home.
Present inventor inhibits cell model and dimethylbenzene inducing mouse ear swelling anxious using inflammatory factor NO generations
Property inflammatory model be anti-inflammatory method of evaluating drug effect.Anti-inflammatory pharmacodynamic evaluation is carried out to said extracted object, the results show that the present invention
Leaf of Longestapex Beautyberry extract all shows preferable anti-inflammatory activity, can be as the application for preparing anti-inflammatory drug or anti-inflammatory ancillary drug.
Beneficial effects of the present invention：
(1) mixing containing three kinds of carnosol, rosmanol and different rosmanol in Leaf of Longestapex Beautyberry extract of the invention
Object, these ingredients are diterpene phenols component, which has preferable anti-oxidant, anticancer and the various actives such as anti-inflammatory.
By the pharmacodynamic study to Leaf of Longestapex Beautyberry extract, broader space will be provided for its further application.
(2) present invention use the similar principle that mixes, in utilization the organic solvent of low polarity by diterpene phenols component rapidly
It is extracted from Leaf of Longestapex Beautyberry, and passes through column chromatography for separation technology, effectively remove depigmentation, be enriched with diterpene phenols component.
(3) preparation method of Leaf of Longestapex Beautyberry extract of the invention is simple, and operation is easy, wide market, is suitble to scale
Metaplasia is produced.
(4) Leaf of Longestapex Beautyberry extract of the invention can be applied to prepare anti-inflammatory drug or anti-inflammatory ancillary drug, have wide
Application space.
Description of the drawings
Fig. 1 is the HPLC measure of three kinds of carnosol in the embodiment of the present invention 1, rosmanol and different rosmanol reference substances
Collection of illustrative plates.
Fig. 2 is that the HPLC for the Leaf of Longestapex Beautyberry extract that the embodiment of the present invention 2 obtains measures collection of illustrative plates.
Fig. 3 is that the HPLC for the Leaf of Longestapex Beautyberry extract that the embodiment of the present invention 3 obtains measures collection of illustrative plates.
Fig. 4 is that the HPLC for the Leaf of Longestapex Beautyberry extract that the embodiment of the present invention 4 obtains measures collection of illustrative plates.
Fig. 5 is that the HPLC for the Leaf of Longestapex Beautyberry extract that the embodiment of the present invention 5 obtains measures collection of illustrative plates.
Fig. 6 is that the HPLC for the Leaf of Longestapex Beautyberry extract that the embodiment of the present invention 6 obtains measures collection of illustrative plates.
Specific embodiment
The principle of the present invention and feature are described below in conjunction with specific attached drawing, example is served only for explaining this hair
It is bright, it is not intended to limit the scope of the present invention.
Embodiment 1
Carnosol 2.0mg, rosmanol 2.2mg and different rosmanol 2.5mg are weighed respectively, and one is placed in 10mL capacity
In bottle, as reference substance, methanol is added to dissolve and is settled to scale to get mixing reference substance prepare liquid, draws the 10 above-mentioned mixing of μ L
Reference substance prepare liquid is injected in efficient liquid phase and is measured.
Liquid phase chromatogram condition：1200 liquid chromatographs of Agilent, chromatographic column：ZORBAX SB-C18(5 μm,
4.6x250mm), column temperature：30 DEG C, flow velocity：1mL/min, Detection wavelength：210nm, mobile phase acetonitrile (A)/water (B)：Gradient is washed
It is de-, 0-25min, 40%-74% (A).
The results are shown in Figure 1.
As shown in Figure 1, under above-mentioned chromatographic condition, separating degree is good for carnosol, rosmanol and different rosmanol.
Embodiment 2
The Leaf of Longestapex Beautyberry extract of the present embodiment contains carnosol, rosmanol and different fan in the Leaf of Longestapex Beautyberry extract
The repeatedly mixture of fragrant phenol, wherein, the mass content of carnosol is 79.6% in the Leaf of Longestapex Beautyberry extract, and the Leaf of Longestapex Beautyberry carries
The mass percentage for taking rosmanol in object is 7.8%, and the quality percentage of different rosmanol contains in the Leaf of Longestapex Beautyberry extract
Measure is 6.6%.
The preparation method of above-mentioned Leaf of Longestapex Beautyberry extract, includes the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
After the stem, branch, leaf of raw material Leaf of Longestapex Beautyberry is taken to dry, it is crushed to obtained by 2mm, obtains dry Leaf of Longestapex Beautyberry powder.
The Leaf of Longestapex Beautyberry powder that 600g is taken to dry adds in the dichloromethane of 10 times of Leaf of Longestapex Beautyberry powder quality, is carried in 50 DEG C of reflux
It takes 3 times, 1 hour every time, crosses 30 μm of sieves, merging filtrate, the proportion that filtrate to 65 DEG C is concentrated under reduced pressure is 1.02 medicinal extract, is obtained
32.2g Leaf of Longestapex Beautyberry crude extract.
Step 2：Purifying
By the Leaf of Longestapex Beautyberry crude extract that 30g steps 1 obtain by silica gel column chromatography purify, first with petroleum ether remove grease type into
Point, then using volume ratio as 5:1 petroleum ether:The mixed solvent of ethyl acetate is eluted, and collects eluent, and recycling design is done
It is dry, obtain the Leaf of Longestapex Beautyberry extract of 10.4g purifying.
It is prepared by the HPLC sample tests liquid of the present embodiment：The above-mentioned Leaf of Longestapex Beautyberry extract 3.6mg being prepared is weighed, is put
Into 10mL volumetric flasks, methanol is added to dissolve and is settled to scale to get HPLC prepare liquids, draws the 10 above-mentioned prepare liquids of μ L, injection
It is measured in efficient liquid phase.
Liquid phase chromatogram condition：With embodiment 1.
The results are shown in Figure 2.
As shown in Figure 2, it is analyzed through HPLC, the mass percentage of carnosic acid is 79.6%.
Application of the above-mentioned Leaf of Longestapex Beautyberry extract in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
Embodiment 3
The Leaf of Longestapex Beautyberry extract of the present embodiment contains carnosol, rosmanol and different fan in the Leaf of Longestapex Beautyberry extract
The repeatedly mixture of fragrant phenol, wherein, the mass content of carnosol is 59.6% in the Leaf of Longestapex Beautyberry extract, and the Leaf of Longestapex Beautyberry carries
The mass percentage for taking rosmanol in object is 12.4%, and the quality percentage of different rosmanol contains in the Leaf of Longestapex Beautyberry extract
Measure is 8.0%.
The preparation method of above-mentioned Leaf of Longestapex Beautyberry extract, includes the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
After the stem, branch, leaf of raw material Leaf of Longestapex Beautyberry is taken to dry, it is crushed to obtained by 2mm, obtains dry Leaf of Longestapex Beautyberry powder.
The Leaf of Longestapex Beautyberry powder that 600g is taken to dry adds in the chloroform of 15 times of Leaf of Longestapex Beautyberry powder quality, is carried in 60 DEG C of reflux
It takes 3 times, 1 hour every time, crosses 50 μm of sieves, merging filtrate, the proportion that filtrate to 65 DEG C is concentrated under reduced pressure is 1.05 medicinal extract, is obtained
34.3g Leaf of Longestapex Beautyberry crude extract.
Step 2：Purifying
The Leaf of Longestapex Beautyberry crude extract that 30g steps 1 obtain is purified through the C18 column chromatographys that grain size is 40-60 μm, with volumetric concentration
Methanol for 40% removes impurity, then is eluted by 75% methanol of volumetric concentration, collects eluent, recycles solution, dry, i.e.,
Obtain the Leaf of Longestapex Beautyberry extract of 4.5g purifying.
It is prepared by the HPLC sample tests liquid of the present embodiment：With embodiment 2.
Liquid phase chromatogram condition：With embodiment 1.
The results are shown in Figure 3.
From the figure 3, it may be seen that being analyzed through HPLC, the mass percentage of carnosic acid is 59.6%.
Application of the above-mentioned Leaf of Longestapex Beautyberry extract in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
Embodiment 4
The Leaf of Longestapex Beautyberry extract of the present embodiment contains carnosol, rosmanol and different fan in the Leaf of Longestapex Beautyberry extract
The repeatedly mixture of fragrant phenol, wherein, the mass content of carnosol is 82.6% in the Leaf of Longestapex Beautyberry extract, and the Leaf of Longestapex Beautyberry carries
The mass percentage for taking rosmanol in object is 5.4%, and the quality percentage of different rosmanol contains in the Leaf of Longestapex Beautyberry extract
Measure is 2.1%.
The preparation method of above-mentioned Leaf of Longestapex Beautyberry extract, includes the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
After the stem, branch, leaf of raw material Leaf of Longestapex Beautyberry is taken to dry, it is crushed to obtained by 2mm, obtains dry Leaf of Longestapex Beautyberry powder.
The Leaf of Longestapex Beautyberry powder that 600g is taken to dry adds in the acetone of 20 times of Leaf of Longestapex Beautyberry powder quality, in 70 DEG C of refluxing extractions 3
It is secondary, 1 hour every time, 70 μm of sieves are crossed, merging filtrate, the proportion that filtrate to 65 DEG C is concentrated under reduced pressure is 1.08 medicinal extract, obtains 51.4g
Leaf of Longestapex Beautyberry crude extract.
Step 2：Purifying
The Leaf of Longestapex Beautyberry crude extract that 30g steps 1 obtain is purified through the MCI column chromatographys that grain size is 75-150 μm, it is dense with volume
The ethyl alcohol spent for 45% removes impurity, then using volumetric concentration as 85% ethanol elution, collects eluent, recycles solution, dry,
Obtain the Leaf of Longestapex Beautyberry extract of 10.4g purifying.
It is prepared by the HPLC sample tests liquid of the present embodiment：With embodiment 2.
Liquid phase chromatogram condition：With embodiment 1.
The results are shown in Figure 4.
As shown in Figure 4, it is analyzed through HPLC, the mass percentage of carnosic acid is 82.6%.
Application of the above-mentioned Leaf of Longestapex Beautyberry extract in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
Embodiment 5
The Leaf of Longestapex Beautyberry extract of the present embodiment contains carnosol, rosmanol and different fan in the Leaf of Longestapex Beautyberry extract
The repeatedly mixture of fragrant phenol, wherein, the mass content of carnosol is 63.2% in the Leaf of Longestapex Beautyberry extract, and the Leaf of Longestapex Beautyberry carries
The mass percentage for taking rosmanol in object is 7.9%, and the quality percentage of different rosmanol contains in the Leaf of Longestapex Beautyberry extract
Measure is 4.9%.
The preparation method of above-mentioned Leaf of Longestapex Beautyberry extract, includes the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
After the stem, branch, leaf of raw material Leaf of Longestapex Beautyberry is taken to dry, it is crushed to obtained by 2mm, obtains dry Leaf of Longestapex Beautyberry powder.
The Leaf of Longestapex Beautyberry powder that 600g is taken to dry adds in the ethyl acetate of 20 times of Leaf of Longestapex Beautyberry powder quality, is carried in 55 DEG C of reflux
It takes 3 times, 1 hour every time, crosses 55 μm of sieves, merging filtrate, the proportion that filtrate to 65 DEG C is concentrated under reduced pressure is 1.02 medicinal extract, is obtained
43.6g Leaf of Longestapex Beautyberry crude extract.
Step 2：Purifying
By the Leaf of Longestapex Beautyberry crude extract that 30g steps 1 obtain by silica gel column chromatography purify, first with petroleum ether remove grease type into
Point, then using volume ratio as 6:1 petroleum ether and the mixed solvent of acetone are eluted, and collect eluent, and recycling design is dry,
Obtain the Leaf of Longestapex Beautyberry extract of 9.3g purifying.
It is prepared by the HPLC sample tests liquid of the present embodiment：With embodiment 2.
Liquid phase chromatogram condition：With embodiment 1.
The results are shown in Figure 5.
As shown in Figure 5, it is analyzed through HPLC, the mass percentage of carnosic acid is 63.2%.
Application of the above-mentioned Leaf of Longestapex Beautyberry extract in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
Embodiment 6
The Leaf of Longestapex Beautyberry extract of the present embodiment contains carnosol, rosmanol and different fan in the Leaf of Longestapex Beautyberry extract
The repeatedly mixture of fragrant phenol, wherein, the mass content of carnosol is 53.0% in the Leaf of Longestapex Beautyberry extract, and the Leaf of Longestapex Beautyberry carries
The mass percentage for taking rosmanol in object is 4.5%, and the quality percentage of different rosmanol contains in the Leaf of Longestapex Beautyberry extract
Measure is 4.4%.
The preparation method of above-mentioned Leaf of Longestapex Beautyberry extract, includes the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
After the stem, branch, leaf of raw material Leaf of Longestapex Beautyberry is taken to dry, it is crushed to obtained by 2mm, obtains dry Leaf of Longestapex Beautyberry powder.
The Leaf of Longestapex Beautyberry powder that 600g is taken to dry adds in the ethyl acetate of 20 times of Leaf of Longestapex Beautyberry powder quality, is carried in 65 DEG C of reflux
It takes 3 times, 1 hour every time, crosses 65 μm of sieves, merging filtrate, the proportion that filtrate to 65 DEG C is concentrated under reduced pressure is 1.06 medicinal extract, is obtained
44.6g Leaf of Longestapex Beautyberry crude extract.
Step 2：Purifying
By the Leaf of Longestapex Beautyberry crude extract that 30g steps 1 obtain by silica gel column chromatography purify, first with petroleum ether remove grease type into
Point, then using volume ratio as 4:1 petroleum ether and the mixed solvent of acetone are eluted, and collect eluent, and recycling design is dry,
Obtain the Leaf of Longestapex Beautyberry extract of 12.8g purifying.
It is prepared by the HPLC sample tests liquid of the present embodiment：With embodiment 2.
Liquid phase chromatogram condition：With embodiment 1.
The results are shown in Figure 6.
It will be appreciated from fig. 6 that being analyzed through HPLC, the mass percentage of carnosic acid is 53.0%.
Application of the above-mentioned Leaf of Longestapex Beautyberry extract in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
NO generates Inhibition test
It takes the logarithm the RAW264.7 cells in growth period, after being counted with cell counter, is made 5 × 104-6 ×104A/mL
Single cell suspension, be inoculated in 12 orifice plates, per hole 1mL, be placed in 37 DEG C, 5%CO2Cell incubator in cultivate.Culture is for 24 hours
Afterwards, it sets respectively and sets dosing group (the Leaf of Longestapex Beautyberry extract that embodiment 2-6 is prepared), LPS groups (a concentration of 0.2 μ g/mL) respectively
And blank control group, wherein dosing group add in the drug of a concentration of 25 μ g/mL, 12.5 μ g/mL, 6.25 μ g/mL, 3.125 μ g/mL
(25 μ g/mL are maximum safe concentration) and the LPS of a concentration of 0.2 μ g/mL, every group sets 3 parallel holes.After continuing culture for 24 hours, receive
Culture medium is taken, Griess methods detect NO contents in culture medium, calculate IC50 values.The results are shown in Table 1.
The influence of RAW264.7 cells release NO that the Leaf of Longestapex Beautyberry extract that 1 embodiment 2-6 of table is prepared induces LPS
As shown in Table 1, the Leaf of Longestapex Beautyberry extract that embodiment 2-6 is prepared shows stronger suppression under safe concentration
NO activity processed.It can be seen that the Leaf of Longestapex Beautyberry extract of the present invention can be as the application for preparing anti-inflammatory drug or anti-inflammatory ancillary drug.
Mice ear is tested
Take KM male mices 70, weight 18-22g.After adaptability is fed 1 week (weight 27-30g at this time), by body
Quality is randomly divided into 7 groups, if model group, positive controls (aspirin) and the experimental group (fine stern that embodiment 2-6 is prepared
Maple extract).By 0.2mL/10g gastric infusions after grouping, positive controls give 200mg/Kg aspirin, experimental group difference
The corresponding drug for giving 400mg/Kg, model group give the distilled water of equivalent.Gavage 1 time/d, continuous 3d, 1h after last gavage,
Dimethylbenzene 0.015mL is uniformly smeared in the forward and backward face of auris dextra of 8 groups of mouse using liquid-transfering gun.1h after smearing, at cervical dislocation
Dead mouse cuts left and right ear along auricle baseline, lays round auricle at the same position of two ears respectively with 7mm card punch, weighs, and calculates
Swelling inhibiting rate.
The results are shown in Table 2.
Influence of the 2 Leaf of Longestapex Beautyberry extract of table to mice ear
As shown in Table 2, model group swelling reaches 15.64mg, and positive aspirin group inhibiting rate illustrates mould up to 33.76%
Type is good.Compared with model group, the Leaf of Longestapex Beautyberry extract that embodiment 2-6 is prepared shows swelling inhibitory activity.
It can be seen that the Leaf of Longestapex Beautyberry extract of the present invention, rich in diterpene phenols component, which has preferable antioxygen
Change, anticancer and the various actives such as anti-inflammatory, will be that its further application carry by the pharmacodynamic study to Leaf of Longestapex Beautyberry extract
For broader space.The Leaf of Longestapex Beautyberry extract of the present invention can be applied to prepare anti-inflammatory drug or anti-inflammatory ancillary drug, have
Wide application space.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of Leaf of Longestapex Beautyberry extract, which is characterized in that contain carnosol, rosmanol and different in the Leaf of Longestapex Beautyberry extract
The mixture of rosmanol, wherein, the mass content of carnosol is 53.0-82.6% in the Leaf of Longestapex Beautyberry extract, described
The mass percentage of rosmanol is 4.5-12.4% in Leaf of Longestapex Beautyberry extract, different rosmanol in the Leaf of Longestapex Beautyberry extract
Mass percentage be 2.1-8.0%.
2. a kind of preparation method of Leaf of Longestapex Beautyberry extract, which is characterized in that include the following steps：
Step 1：Prepare Leaf of Longestapex Beautyberry crude extract
Dry Leaf of Longestapex Beautyberry powder is taken, the organic solvent refluxing extraction of 10-20 times of addition Leaf of Longestapex Beautyberry powder quality 3 times is filtered, closed
And filtrate, filtrate is concentrated under reduced pressure and obtains medicinal extract, as Leaf of Longestapex Beautyberry crude extract；
Step 2：Purifying
The Leaf of Longestapex Beautyberry crude extract that step 1 obtains is purified by silica gel column chromatography, first with petroleum ether removal grease-like composition, then with
Volume ratio is (5-3):1 petroleum ether:The mixed solvent or volume ratio of ethyl acetate are (6-4):1 petroleum ether and acetone it is mixed
Bonding solvent is eluted, and collects eluent, recycling design, dry to get to the Leaf of Longestapex Beautyberry extract of purifying；
Or the inverted column of material chromatographic purifying of Leaf of Longestapex Beautyberry crude extract for obtaining step 1, using volumetric concentration as the alcohol of 40-50%
Class removes impurity, then the alcohols using volumetric concentration as 75-95% elutes, and collects eluent, recycles solution, dry to get to pure
The Leaf of Longestapex Beautyberry extract of change.
A kind of 3. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that fine stern described in step 1
Maple powder is after taking stem, branch, the leaf drying of raw material Leaf of Longestapex Beautyberry, to be crushed to obtained by 2mm.
4. the preparation method of a kind of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that organic described in step 1
Solvent is dichloromethane, one kind in chloroform, ethyl acetate, acetone.
A kind of 5. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that extraction described in step 1
Temperature be 50-70 DEG C, every time 1 hour.
A kind of 6. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that filtering described in step 1
Aperture be 30-70 μm.
A kind of 7. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that medicinal extract described in step 1
It is 1.02-1.08 in 65 DEG C of proportions.
A kind of 8. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that reverse phase described in step 2
The medium of column chromatography is one kind in C18, C8, C4 or MCI.
A kind of 9. preparation method of Leaf of Longestapex Beautyberry extract according to claim 2, which is characterized in that alcohols described in step 2
It is methanol or ethyl alcohol.
10. a kind of application of Leaf of Longestapex Beautyberry extract as described in claim 1 in anti-inflammatory drug or anti-inflammatory ancillary drug is prepared.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115721775A (en) * | 2022-12-09 | 2023-03-03 | 海南苏生生物科技有限公司 | Composite modified biological hydrogel dressing and preparation process thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN115721775A (en) * | 2022-12-09 | 2023-03-03 | 海南苏生生物科技有限公司 | Composite modified biological hydrogel dressing and preparation process thereof |
| CN115721775B (en) * | 2022-12-09 | 2023-09-29 | 海南苏生生物科技有限公司 | Composite modified biological hydrogel dressing and preparation process thereof |
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