CN103751792B - Radix Angelicae Sinensis extract is as the application of natural intestinal absorption enhancer - Google Patents

Radix Angelicae Sinensis extract is as the application of natural intestinal absorption enhancer Download PDF

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CN103751792B
CN103751792B CN201410017214.XA CN201410017214A CN103751792B CN 103751792 B CN103751792 B CN 103751792B CN 201410017214 A CN201410017214 A CN 201410017214A CN 103751792 B CN103751792 B CN 103751792B
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angelicae sinensis
radix angelicae
intestinal
medicine
sinensis extract
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CN103751792A (en
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易军
王晓明
张凤玲
黄燕芬
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Guangdong Pharmaceutical University
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Abstract

The present invention relates to medical art, disclose the application of Radix Angelicae Sinensis extract as natural intestinal absorption enhancer.The present invention, using Radix Angelicae Sinensis extract as natural intestinal absorption enhancer, can use with being promoted compatibility of drugs or making composite drug, can play significant cooperative effect, improves the bioavailability being promoted medicine, and can not cause damage to the mucosa of absorption site.Radix Angelicae Sinensis extract is applied as natural intestinal absorption enhancer and has major application value.

Description

Radix Angelicae Sinensis extract is as the application of natural intestinal absorption enhancer
Technical field
The present invention relates to medical art, more specifically, relate to the application of Radix Angelicae Sinensis extract as natural intestinal absorption enhancer.
Background technology
Research finds, in the research of medicine, is eliminated or needs to be improved by preparation means to absorb after having the noval chemical compound of 39% oral because of gastrointestinal absorption problem.There is absorption problem equally in Chinese medicine.Compared with chemicals, the mechanism of absorption of Chinese medicine or Effective Component of Chinese Medicine is more indefinite.Motherland's Chinese medicine is our rarity, but how Chinese medicine enters blood by the intestines and stomach is the problem that Chinese medicine and pharmacy worker is most interested in, but progress at present in described problem little.
Intestinal permeation enhancers refers to the material that medicine can be promoted at gastrointestinal tract transmembrane transport, and its Main Function mechanism comprises: mobility, the promotion Cell membrane pores of (1) enhancing cell membrane are formed, reduce mucous layer viscosity, improve membrane permeability; (2) suppress hydrolytic enzyme effect, strengthen medicine effect capability development; (3) temporarily change the tight connection between agents area epithelial cell, be beneficial to medicine and pass through; (4) prevent protein aggregation, strengthen the thermodynamic motion of medicine; (5) increase agents area unit interval blood flow, improve the gradient of drug level inside and outside cell membrane.
Current intestinal permeation enhancers mainly contains surfactant-based, cholate, fatty acid and fatty acid salt, bioadhesive high molecular polymer class etc.
Radix Angelicae Sinensis ( angelica sinensis), herbaceos perennial, be distributed in the ground such as Gansu, Yunnan, Sichuan, Qinghai, Shaanxi, Hunan, Hubei, Guizhou in China, all there is cultivation various places.The root of Radix Angelicae Sinensis can be used as medicine, and is one of the most frequently used Chinese medicine.Radix Angelicae Sinensis also can be used in spiced and stewed food batching, and its main feature is flavor and enhancing flavor, increases meat products and medicated incense taste.Radix Angelicae Sinensis can form medicine pair with plurality of Chinese compatibility, but, drug matching and the right Forming Mechanism more complicated of medicine, the concrete mechanism that existing research does not provide Radix Angelicae Sinensis to promote drug absorption and embody rule effect, the indefinite development and application affecting medicine the most at last of mechanism and effect.The conventional Radix Angelicae Sinensis extract adopting the methods such as water extraction, aqueous alkali lifting manipulation, alcohol dipping method, alcohol heat reflux method, alcohol percolation method to prepare, containing volatile oil composition and organic acid, is mainly succinic acid, ligustilide, ferulic acid etc.
Have no Radix Angelicae Sinensis extract at present to report as the technology of related drugs intestinal permeation enhancers.
Summary of the invention
The technical problem to be solved in the present invention is the deficiency overcoming existing natural intestinal absorption enhancer application technology, provides Radix Angelicae Sinensis extract as the application of natural intestinal absorption enhancer.
Object of the present invention is achieved by the following technical programs:
There is provided Radix Angelicae Sinensis extract as the application of the natural intestinal absorption enhancer of medicine.
Described medicine include but not limited to duodenum, jejunum, colon and/or ileum site absorption medicine especially duodenal site absorb medicine.
As one of preferred application scheme, described Radix Angelicae Sinensis extract and compatibility of drugs are applied.
As preferred application scheme two, described Radix Angelicae Sinensis extract and medicine are prepared jointly to be become medicament and applies.
The active component of Radix Angelicae Sinensis extract to a lot of medicine and medicine has significant intestinal absorption facilitation, and more preferably, Radix Angelicae Sinensis extract has significant intestinal absorption facilitation to astragaloside, gastrodine or peoniflorin etc.
Described Radix Angelicae Sinensis extract and medicine are prepared jointly to be become medicament and applies, and preferred dosage form is oral administered dosage form.Include but not limited to as tablet, capsule, granule, oral powder, oral solution, Orally taken emulsion or oral suspensions.
The present invention further provides active component ferulic acid in Radix Angelicae Sinensis extract to apply as the natural intestinal absorption enhancer of medicine.
Preferably, Radix Angelicae Sinensis extract active component ferulic acid significantly can promote that medicine can't cause damage to absorption site in the absorption of duodenal site.
Radix Angelicae Sinensis extract active component ferulic acid significantly can promote that astragaloside is in the absorption of duodenal site.And be realize penetration enhancement effect by improving duodenal absorption rate constant Ka and apparent absorption coefficient Papp.
Beneficial effect of the present invention:
The invention provides the application of Radix Angelicae Sinensis extract as natural intestinal absorption enhancer, described Radix Angelicae Sinensis extract has good intestinal absorption facilitation, and the medicine be promoted enters after in human body, can play cooperative effect, improves the bioavailability of medicine.The application of described Radix Angelicae Sinensis extract, can reduce the dosage of medicine and the use of other intestinal permeation enhancers, reduces the toxic and side effects of medicine and the absorption of other ineffective treatments foreign body.
Research summary of the present invention Radix Angelicae Sinensis extract has significant intestinal absorption facilitation to related drugs, is inquire into the compatibility principle that Radix Angelicae Sinensis and other Chinese medicine forms medicine right to provide strong technical foundation from the angle of intestinal absorption.
Further, the active component ferulic acid that the invention provides Radix Angelicae Sinensis extract is the application of astragaloside intestinal permeation enhancers, ferulic acid has good intestinal absorption facilitation to astragaloside, enter after in human body with astragaloside, cooperative effect can be played, promote that astragaloside is in intestinal absorption effect, improve blood drug level, the toxic and side effects of medicine and the absorption of other ineffective treatments foreign body can be reduced, damage can not be caused to intestinal absorption position mucosa, make full use of ferulic acid, be used as astragaloside intestinal permeation enhancers and there is major application value.
Accompanying drawing explanation
Fig. 1 is that different absorption enhancer is urged to ooze ratio at duodenal Ka to peoniflorin;
Fig. 2 is that absorption enhancer compares at duodenum Papp peoniflorin;
Fig. 3 be ferulic acid astragaloside is absorbed at different intestinal segment impact ( ± s, n=5);
Fig. 4 is the promotion of ferulic acid to astragaloside intestinal transport Ka;
Fig. 5 is the promotion of ferulic acid to astragaloside intestinal transport Papp;
Fig. 6 is rat preduodenal intestine colibacillosis (HE dyes, 400 times).
Detailed description of the invention
The present invention is further illustrated below in conjunction with the drawings and specific embodiments.Unless stated otherwise, the raw material adopted in the embodiment of the present invention and method are the conventional commercial raw material in this area and the conventional method used.
Embodiment 1 Radix Angelicae Sinensis extract is tested the intestinal penetration enhancement effect of peoniflorin
The Radix Angelicae Sinensis extract that the present embodiment adopts is purchased from Ze Lang Zhi Ti bio tech ltd, Nanjing; Peoniflorin to come biological engineering company limited purchased from Xi'an, but does not therefore limit the present invention.Those skilled in the art can adopt other Radix Angelicae Sinensis extract of originating and peoniflorins, also can adopt conventional extracting method self-control.
Utilize rat at the unidirectional intestinal perfusion model of body, adopt HPLC method to measure paeoniflorin content in perfusate, investigate Radix Angelicae Sinensis extract respectively to the facilitation of peoniflorin intestinal absorption.
The test of rat at the unidirectional intestinal perfusion model of body is carried out according to following steps method:
S1. Rat Fast 12 h, freely drinks water, 1% Nembutal sodium solution (5 mL/kg) intraperitoneal injection of anesthesia, and back of the body position is fixed, and keeps body temperature;
S2. open abdominal cavity along ventrimeson and be about 3cm, get duodenum about 10 cm, after being in two ends otch apart from pylorus 1 cm, intubate ligation, intestinal contents is rinsed well with 5 mL/min flow velocitys with the normal saline of 37 DEG C of constant temperature, then with air, normal saline is emptying, erecting device;
S3. with the perfusate being preheated to 37 DEG C with 0.2 mL/min flow velocity perfusion 30 minutes, equalizing line and intestinal segment;
S4. start timing, collect 0-15 respectively, the perfusate of 15-30,30-45,45-60,60-75,75-90 min, weighs, record perfusate and collection liquid weight;
S5., at the end of experiment, intestinal segment length and internal diameter is measured.
Measure the content of peoniflorin in perfusate with HPLC method, and be calculated as follows the absorption rate constant Ka and apparent absorption coefficient Papp that calculate medicine:
Wherein, Q inand Q outbe respectively the perfusate that intestinal segment pours into and the volume (mL) collecting liquid, l and r is respectively length (cm) and the cross sectional radius (cm) of perfusion intestinal segment, and Q is perfusion rate, C inand C outbe respectively perfusate and collect liquid concentration (μ g/mL), V is the volume (cm of perfusion intestinal segment 3).
With the absorption rate constant Ka of medicine and apparent absorption coefficient Papp for index, experimental result shows, HP-β-CD and Radix Angelicae Sinensis extract significantly improve peoniflorin at the absorption rate constant of rat preduodenal and apparent absorption coefficient, show their in the oral formulations of peoniflorin as absorption enhancer use may.
Table 1 absorption enhancer is on the impact of peoniflorin at duodenum Ka
Table 2 absorption enhancer is on the impact of peoniflorin at duodenum Papp
Method same as described above is adopted to test, with absorption rate constant Ka and apparent absorption coefficient Papp for index, the short intestinal absorption effect of Radix Angelicae Sinensis extract to medicines such as astragaloside, gastrodine, peoniflorins is all better than the medicine not adding Radix Angelicae Sinensis extract, and is better than Capric acid sodium salt and the sodium deoxycholate of respective concentration.
Embodiment 2
Laboratory animal: SD rat, male and female half and half, body weight 160 ~ 220 g, Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center provides, the quality certification number: SCXK(Guangdong) 2008-0020.
Instrument: SHIMADZU high performance liquid chromatograph (Japan); TGL-20B high speed centrifuge (Anting Scientific Instrument Factory, Shanghai).
Medicine and reagent: astragaloside reference substance (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 110781-200613); Acetonitrile, methanol (Di Ma company, chromatographically pure); Water is distilled water.Other reagent are analytical pure.
Krebs-Ringer nutritional solution: hereinafter referred to as K-R liquid, every 1000 ml are containing NaCl 7.8g, KCl 0.35g, NaHCO 31.37g, NaH 2pO 40.32g, MgCl 20.02g, glucose 1.4g, supplies 1000mL with water; Face with preparation, the same day uses.
The present embodiment ferulic acid is purchased from Ze Lang Zhi Ti bio tech ltd, Nanjing (lot number 20110903), but therefore do not limit the present invention, those skilled in the art can adopt other ferulic acids of originating, the ferulic acid that the Radix Angelicae Sinensis extract prepared also can be adopted to carry out separating-purifying obtain.Radix Astragali extract can conventional commercial or self-control (containing astragaloside 13.9%).Homemade method is with reference to the following is: get dry astragalus root raw material, cleaning, dry, 10 times amount 70% ethanol are added, heating and refluxing extraction 2 times, each 3 hours after pulverizing, merge secondary raffinate, decompression recycling ethanol, residue adds suitable quantity of water and dissolves, making containing astragaloside concentration is 0.8 ~ 1.1mg/mL, gets D101 macroporous adsorbent resin, with 2 BV/h loadings, be washed to without Molish reaction, with 5 BV 70% ethanol elutions, collect eluent, decompression recycling ethanol, dry, to obtain final product.
1. everted intestinal sac drug solution preparing
Radix Astragali group: get Radix Astragali extract appropriate, dissolve with Krebs-Ringer nutritional solution and be settled to 100 mL, obtained containing astragaloside 34.75 μ gmL -1solution.
Compatibility group: by the dosage (the crude drug mass ratio of Radix Angelicae Sinensis and the Radix Astragali is 1:1) of Radix Angelicae Sinensis-Radix Astragali 1:1, with reference to Radix Astragali group concentration, obtains with method.
2. everted intestinal sac experiment
(Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center provides to get healthy SD rat, the quality certification number: SCXK(Guangdong) 2008-0020) overnight fasting (freely drinking water), by 400 mgkg-1 lumbar injection 1% Nembutal sodium solution anesthesia, divide and get different intestinal segment, intestinal is overturn, intestinal capsule Inner injects the blank Krebs-Ringer nutritional solution of 1 mL, this section of intestinal capsule is put in the everted intestinal sac medicinal liquid of preheating, continue to pass into carbon dioxide: the mist of oxygen (5:95) in medicinal liquid, 37 DEG C of insulation 0.5 h.After experiment terminates, collect serosal fluid, measure the content of its Chinese medicine, and measure intestinal absorption area, investigate the absorption of different intestinal segment to astragaloside respectively, and the impact that ferulic acid absorbs astragaloside.
3. the mensuration of everted intestinal sac serosal fluid drug content
Chromatographic column: Diamonsil C18 post (4.6 mm × 250 mm, 5 μm); Mobile phase: acetonitrile-water (32.4:67.6); Column temperature: 25 DEG C; Flow velocity 1.0 mLmL -1; Sample size: 20 μ L.The specificity of investigation method, result shows, the mensuration of blank intestinal juice to astragaloside is noiseless.Get astragaloside reference substance storing solution appropriate, add the blank intestinal juice of everted intestinal sac, compound concentration is 8.03,16.06,24.06,32.12,40.15,48.18,56.21 μ gmL -1the solution of series concentration, returns astragaloside concentration (C) peak area (A), obtains standard curve: A=716.8C – 108.53, R=0.9994, the range of linearity: 8.03 ~ 56.21 μ gmL-1.Lowest detectable limit is 1.5 μ gmL -1.The response rate of high, normal, basic three concentration is 93.9% ~ 97.4%, RSD is 1.40% ~ 2.81%, and withinday precision RSD is 0.54% ~ 0.78%, and day to day precision RSD is 0.58% ~ 0.99%, all meets the requirements.
4. experimental result
Adopt the impact that the rat everted intestinal sac model investigation Radix Astragali absorbs it at absorption and the ferulic acid of different intestinal segment, result shows, astragaloside all has absorption at the whole intestinal segment of small intestinal, different intestinal segment Papp sequence is: duodenum > jejunum > colon > ileum, wherein duodenal absorption is best.After adding ferulic acid, the absorption increase of duodenum and jejunum is comparatively obvious, sees shown in accompanying drawing 3.
5. experiment conclusion
In everted intestinal sac experiment, Zhu Jingyun etc. are active with determination of glucose oxidase intestinal capsule, confirm that in vitro intestinal capsule activity can only maintain 1h, the present invention determines the drug absorption transhipment of time period (0.5h) in everted intestinal sac experiment, and by constant temperature 37 DEG C, pass into oxygen-enriched air, ensure that data are true and reliable.Experimental summary of the present invention finds, ferulic acid can promote the absorption of astragaloside well, especially promotes that astragaloside is in duodenal absorption.
Embodiment 3
1. at body unidirectional intestinal perfusion drug solution preparing
Radix Astragali group: get Radix Astragali extract appropriate, dissolve and be settled to 100 mL with sodium chloride injection, preparation astragaloside concentration is respectively 10.5,15.75,21.00,31.50,36.75 μ gmL -1the perfusate of totally five gradient concentrations.
Compatibility group: by the dosage (the crude drug mass ratio of Radix Angelicae Sinensis and the Radix Astragali is 1:1) of Radix Angelicae Sinensis-Radix Astragali 1:1, with reference to Radix Astragali group concentration, obtains with method.
2. at the unidirectional intestinal perfusion experiment of body
Get healthy SD rat (Traditional Chinese Medicine University Of Guangzhou's Experimental Animal Center provides, the quality certification number: SCXK(Guangdong) 2008-0020) overnight fasting (freely drinking water), by 400 mgkg -1lumbar injection 1% Nembutal sodium solution is anaesthetized, and opens abdominal cavity, and to investigation intestinal segment (getting about 10 cm) two ends otch intubate, ligation, forms closed circuit.With the normal saline of 37 DEG C, intestinal contents is rinsed well, get 37 DEG C at body unidirectional intestinal perfusion medicinal liquid 100 mL, first with 1.0 mLmin -1flow velocity perfusion 10 min, then flow velocity is adjusted to 0.2 mLmin -1, after pre-equilibration 30 min, collect the perfusate of 30 ~ 45,45 ~ 60,60 ~ 75,75 ~ 90,90 ~ 105,105 ~ 120 min time periods and weigh.After experiment terminates, cut intestinal segment, measure absorption area.Measure perfusate concentration, investigate the impact that astragaloside absorbs astragaloside at absorption and the ferulic acid of this intestinal segment respectively.
3. in the mensuration of body intestinal perfusate Chinese medicine
By chromatographic condition under " mensuration of everted intestinal sac serosal fluid drug content " item, the specificity of investigation method, result shows, in intestinal perfusate, impurity does not disturb the mensuration of astragaloside.Get astragaloside reference substance appropriate, be added in the blank intestinal juice of body intestinal perfusion, compound concentration is 8.71,17.42,26.13,34.84,43.55,52.26,60.97 μ gmL -1the solution of series concentration, centrifugal 10 min of 12000 rpm, get supernatant, with the filtering with microporous membrane of 0.45 μm, get filtrate 20 μ L sample introduction to measure, astragaloside concentration (C) is returned peak area (A), obtains standard curve: A=829.3C-76.24, R=0.9995, the range of linearity: 8.71 ~ 60.97 μ gmL -1.Lowest detectable limit is 1.7 μ gmL -1.The response rate of high, normal, basic three concentration is 92.9% ~ 99.3%, RSD is 1.62% ~ 3.53%, and withinday precision 0.64% ~ 1. 86%, day to day precision 0.38% ~ 1.99%, all meets the requirements.
4. the calculating of drug absorption rate constant Ka and medicine apparent absorption coefficient Papp adopts gravimetric method to flow into perfusate and the volume that flows out corrects, eliminate the impact of its change in volume, calculate absorption rate constant Ka and medicine apparent absorption coefficient Papp by following equation:
In above formula: C inand C outbe respectively intestinal and import and export perfusate drug level (μ gmL -1); Q inand Q outbe respectively the volume (mL) that intestinal imports and exports perfusate; L and r is respectively perfusion intestinal segment length (cm) and cross sectional radius (cm); For perfusion rate (mLmin -1); For the volume (cm of perfusion intestinal segment 3).
5. morphocytology inspection
Be taken at the duodenum 1cm before and after body intestinal perfusion experiment respectively, the tissue slice made with HE staining, carries out the morphological examination of epithelium layer, and whether inspection duodenal epithelium cell keeps active upon administration.
6. experimental result
Choose rat preduodenal, adopt at the unidirectional intestinal perfusion model of body, investigate the astragaloside absorbing state of variable concentrations, and investigate the short Absorption of ferulic acid to astragaloside, the results are shown in shown in accompanying drawing 4, accompanying drawing 5, and table 3, shown in table 4.
Table 3 variable concentrations astragaloside compares ER at the absorption water molecules Ka of rat preduodenal with short oozing
Note: compared with Radix Astragali group, compatibility group all has significant difference, * P<0.05.
Table 4 variable concentrations astragaloside compares ER at the apparent permeability coefficients Papp of rat preduodenal with short oozing
Result shows, the astragaloside of variable concentrations has absorption at duodenum, and after adding ferulic acid, Ka and Papp all increases.
7 morphocytology inspections
Intestinal villi after administration is substantially complete, without obviously damaging, compared with before administration, being substantially consistent, seeing shown in accompanying drawing 6.Prove that the compatibility of astragaloside and ferulic acid of the present invention and astragaloside uses not damage intestinal segment, experimental session intestinal keeps absorbing activity.
8. experiment conclusion
In the unidirectional intestinal perfusion experiment of body, the Radix Astragali, in duodenal absorption, shows as and raises with concentration and increase, show the feature of Passive intake under middle low concentration; But at higher concentrations, Ka and Papp reduces all to some extent, point out the possibility that the intestinal transport of astragaloside has carrier to participate in.
Under the effect of ferulic acid, Ka and the Papp of astragaloside increases all to some extent, and ER value is all greater than 1, and there is significant difference (P<0.05); Wherein with middle concentration (21.00 μ gmL -1) short assimilation effect the most obvious: ERKa=1.42, ERPapp=1.47, it is relevant with astragaloside concentration that the display intestinal of ferulic acid to the Radix Astragali urgees absorbability.

Claims (8)

1. the Radix Angelicae Sinensis extract containing ferulic acid is preparing as the natural intestinal absorption enhancer of medicine astragaloside, gastrodine or peoniflorin the application had in the pharmaceutical preparation of the intestinal absorption of improvement.
2. application according to claim 1, it is characterized in that, the described Radix Angelicae Sinensis extract containing ferulic acid is preparing as the absorption enhancer at the duodenum of medicine astragaloside, gastrodine or peoniflorin, jejunum, colon or ileum one of them or several position the application had in the pharmaceutical preparation of the intestinal absorption of improvement.
3. application according to claim 2, it is characterized in that, the described Radix Angelicae Sinensis extract containing ferulic acid is preparing as the duodenal absorption enhancer of medicine astragaloside, gastrodine or peoniflorin the application had in the pharmaceutical preparation of the intestinal absorption of improvement.
4. application according to claim 1, is characterized in that, the described Radix Angelicae Sinensis extract containing ferulic acid and compatibility of drugs are applied.
5. application according to claim 1, is characterized in that, described Radix Angelicae Sinensis extract containing ferulic acid and medicine are prepared jointly to be become medicament and apply.
6. application according to claim 5, is characterized in that, described medicament is oral administered dosage form.
7. application according to claim 6, is characterized in that, described oral administered dosage form is tablet, capsule, granule, oral powder, oral solution, Orally taken emulsion or oral suspensions.
8. application according to claim 1, is characterized in that, described medicine is astragaloside.
CN201410017214.XA 2014-01-15 2014-01-15 Radix Angelicae Sinensis extract is as the application of natural intestinal absorption enhancer Expired - Fee Related CN103751792B (en)

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Granted publication date: 20151028