CN101513388A - Icariin microemulsion and preparation method thereof - Google Patents
Icariin microemulsion and preparation method thereof Download PDFInfo
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- CN101513388A CN101513388A CNA2009100299281A CN200910029928A CN101513388A CN 101513388 A CN101513388 A CN 101513388A CN A2009100299281 A CNA2009100299281 A CN A2009100299281A CN 200910029928 A CN200910029928 A CN 200910029928A CN 101513388 A CN101513388 A CN 101513388A
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Abstract
The invention relates to an icariin microemulsion which takes icariin as active ingredient; the icariin is dissolved in oil and then added with emulsifying agent, assistant emulsifier and water to prepare microemulsion with the micro emulsion grain diameter of 10-100nm, wherein, the oil can be one or more in oleic acid, ethyl oleate, isopropyl myristate or castor oil; the icariin microemulsion has the components with the mass percent: 0.66-6.02% of saturated solution of oil of icariin, 10-50% of emulsifying agent, 10-38% of assistant emulsifier and the balance water, Hank's balanced salt solution (HBSS), PBS buffer solution or normal saline. The icariin microemulsion can be kept clear, transparent, stable and uniform when being placed at room temperature for a year, thus good stability is verified evidently. The experiments of cells and animals prove that the icariin microemulsion can strengthen the icariin to penetrate cell layers, promote the medicine to be ingested and absorbed and enhance the healing effect of the medicine. The invention also discloses a preparation method of the icariin microemulsion.
Description
Technical field:
The invention belongs to the nanosecond medical science field, particularly a kind of vegetalitas, water-insoluble drug epimedium aglucone are processed into physicochemical property stable microemulsion and preparation method thereof.
Background technology:
Osteoporosis is a worldwide disease, and along with society's aging day by day, osteoporosis has been classified one of three big Senile disease as.Epimedium aglucone is the flavone monomer component that abstraction reaction obtains from traditional Chinese medical science clinical commonly used drug Herba Epimedii.Domestic and international many studies show that: epimedium aglucone has tangible osteoporosis pharmacology activity (referring to Jian Huang, Lan Yuan, Xi Wang, et al.Icaritin and its glycosides enhance osteobIastic, but suppress osteocIastic, differentiation and activity in vitro[J] .Life Sciences, 2007,81:832-840.).But its water solublity is very poor, influences the performance of its oral absorption and drug effect.
Summary of the invention:
Technical problem to be solved by this invention provides that a kind of technology is simple, physical property is stable, and can significantly improve water miscible microemulsion of epimedium aglucone and preparation method thereof.
The technical solution used in the present invention is:
A kind of icariin microemulsion, it is to be active component with the epimedium aglucone, it is dissolved in the oils and fats, add that the microemulsion particle diameter that emulsifying agent, co-emulsifier and water are prepared from is the microemulsion of 10～100nm, described oils and fats can be one or more in oleic acid, ethyl oleate, isopropyl myristate or the Semen Ricini wet goods, wherein, the mass percent of each component is:
The saturated solution 0.66～6.02% of the oil of epimedium aglucone
Co-emulsifier 10～38%
Surplus is water, Hank ' s balanced salt solution (HBSS), PBS buffer or normal saline.
In the above-mentioned microemulsion, described emulsifying agent can be OP.
In the above-mentioned microemulsion, described co-emulsifier can be glycerol, isopropyl alcohol, Macrogol 200, Liquid Macrogol, PEG400 or 1, the 2-propylene glycol.
A kind of method for making of above-mentioned icariin microemulsion, it comprises the steps:
The preparation of step 1. epimedium aglucone oil solution: excessive epimedium aglucone is dissolved in the oil, mixes to make abundant dissolving, centrifugally remove undissolved epimedium aglucone, the saturated solution of oil of epimedium aglucone;
The dissolubility of epimedium aglucone in oil
Step 3. stirs four kinds of components that step 2 took by weighing and makes abundant mixing, gets the icariin microemulsion of clear.
Be terminal point to obtain clear solution in the microemulsion preparation process.The particle diameter of microemulsion is little, good stability.
The icariin microemulsion of the present invention's preparation, room temperature are placed average annual clarification, transparent, stable, the homogeneous of keeping, and fully show its good stable.
The present invention prepares in the epimedium aglucone microemulsion process and does not need to disperse with colloid mill, does not also need to carry out high speed shear (general microemulsion manufacturing all needs), but simple agitation or spontaneous can formation.
Cell and zoopery show: the epimedium aglucone microemulsion can increase epimedium aglucone penetration cell layer, promotes ingestion of medicines to absorb medicament curative effect enhancement.
Description of drawings
Fig. 1 is the particle size distribution figure of the icariin microemulsion of embodiment 8 preparations;
Fig. 2 is the icariin microemulsion Zeta potential scattergram of embodiment 8 preparations;
Fig. 3 is for being the particle size distribution figure of the icariin microemulsion of embodiment 9 preparations;
Fig. 4 is the icariin microemulsion Zeta potential scattergram of embodiment 9 preparations;
Fig. 5 is for being the particle size distribution figure of the icariin microemulsion of embodiment 10 preparations;
Fig. 6 is the icariin microemulsion Zeta potential scattergram of embodiment 10 preparations;
Fig. 7 is for being the particle size distribution figure of the icariin microemulsion of embodiment 11 preparations;
Fig. 8 is the icariin microemulsion Zeta potential scattergram of embodiment 11 preparations;
Fig. 9 is for being the particle size distribution figure of the icariin microemulsion of embodiment 14 preparations;
Figure 10 is the icariin microemulsion Zeta potential scattergram of embodiment 14 preparations;
Figure 11 is for being the particle size distribution figure of the icariin microemulsion of embodiment 15 preparations;
Figure 12 is the icariin microemulsion Zeta potential scattergram of embodiment 15 preparations;
Figure 13 is for being the particle size distribution figure of the icariin microemulsion of embodiment 16 preparations;
Figure 14 is the icariin microemulsion Zeta potential scattergram of embodiment 16 preparations;
Figure 15 is for being the particle size distribution figure of the icariin microemulsion of embodiment 17 preparations;
Figure 16 is the icariin microemulsion Zeta potential scattergram of embodiment 17 preparations;
Figure 17 is for being the particle size distribution figure of the icariin microemulsion of embodiment 18 preparations;
Figure 18 is the icariin microemulsion Zeta potential scattergram of embodiment 18 preparations;
Figure 19 is for being the particle size distribution figure of the icariin microemulsion of embodiment 19 preparations;
Figure 20 is the icariin microemulsion Zeta potential scattergram of embodiment 19 preparations;
Figure 21 is embodiment 22 icariin microemulsion anti-osteoporosis activity test patterns, and a is blank group; B is the icariin microemulsion group.
The specific embodiment
Further specify the present invention below by some examples, but do not limit the present invention in the described example ranges.
The preparation of embodiment 1. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 2. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 3. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 4. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 5. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 6. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the isopropyl myristate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 7. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the isopropyl myristate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 8. icariin microemulsions
Take by weighing OP, isopropyl alcohol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Fig. 1 and Fig. 2.
The preparation of embodiment 9. icariin microemulsions
Take by weighing OP, PEG400 by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Fig. 3 and Fig. 4.
The preparation of embodiment 10. icariin microemulsions
Take by weighing OP, Liquid Macrogol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Fig. 5 and Fig. 6.
The preparation of embodiment 11. icariin microemulsions
Take by weighing OP, Macrogol 200 by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Fig. 7 and Fig. 8.
Embodiment 12
Take by weighing OP, isopropyl alcohol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
Embodiment 13
Take by weighing OP, isopropyl alcohol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.
The preparation of embodiment 14. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the Oleum Ricini saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Fig. 9 and Figure 10.
The preparation of embodiment 15. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds normal saline, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Figure 11 and Figure 12.
The preparation of embodiment 16. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds the PBS buffer, and mix homogeneously adds the ethyl oleate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Figure 13 and Figure 14.
The preparation of embodiment 17. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds HBSS, and mix homogeneously adds the isopropyl myristate saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Figure 15 and Figure 16.
The preparation of embodiment 18. icariin microemulsions
Take by weighing OP, glycerol by following prescription, mix homogeneously adds entry, and mix homogeneously adds the oleic acid saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Figure 17 and Figure 18.
The preparation of embodiment 19. icariin microemulsions
Take by weighing OP, 1 by following prescription, the 2-propylene glycol, mix homogeneously adds entry, and mix homogeneously adds the oleic acid saturated solution of epimedium aglucone again, stirs to clarify transparent.The particle size distribution of icariin microemulsion and Zeta potential are seen Figure 19 and Figure 20.
Investigate the absorption and transport situation of epimedium aglucone microemulsion with the Caco-2 cell model:
The foundation of Caco-2 cell monolayer model:
Caco-2 cell monolayer model places the 75mL culture bottle, is culture fluid with DMEM.At 37 ℃, 5%CO
2Cultivate in the incubator, changed 1 culture fluid, cultivated 5～7 days, treat that cell covers with culture bottle, with containing 0.25% tryptic Digestive system digestion every 1 day.General digestion time is about 5～8 minutes.The microscopically counting.Behind the centrifugal 5min of 3000r/min, be suspended in the culture fluid, be seeded on the Transwells that usefulness Mus tail collagen was handled in advance, cell inoculation density is 100,000 cell cm
-2The Caco-2 cell was differentiated to form in about 19～21 days, promptly can be used for test.
The transhipment process of the test:
37 ℃ of HBSS that use pH 7.4 down measure cell monolayer flushing 3 times to stride membrane resistance, discard and stride the membrane resistance value less than 500 Ω * cm
2Cell.Cell siphons away incubation medium hatch 1h in buffer after.
For the transhipment of medicine: add to contain at the villous surface (AP) of cellular layer and remain the solution of reagent thing as supply pool from cell villous surface Apical (A) side to basal surface Basolateral (B) side, add barren HBSS solution as reception tank at basal surface (BL), a series of transmembrane transport takes place subsequently.The liquid 400 μ L that in 60min, take a sample at the bottom of the AP side, and replenish and contain the solution for the treatment of the reagent thing with volume and keep the AP Side Volume constant; The liquid 400 μ L that in 60min, take a sample at the bottom of the BL side, and replenish with liquid of the blank end of volume and keep the BL Side Volume constant.Add 100 μ L solution in every duplicate samples, mixture is centrifugal, and supernatant HPLC analyzes.
For the transhipment of medicine: add to contain at the basal surface (BL) of cellular layer and remain the solution of reagent thing as supply pool from basal surface Basolateral (B) side to cell villous surface Apical (A) side, add barren HBSS solution as reception tank at villous surface (AP), a series of transmembrane transport takes place subsequently.The liquid 400 μ L that in 60min, take a sample at the bottom of the BL side, and replenish and contain the solution for the treatment of the reagent thing with volume and keep the BL Side Volume constant; The liquid 400 μ L that in 60min, take a sample at the bottom of the AP side, and replenish with liquid of the blank end of volume and keep the AP Side Volume constant.Add 100 μ L solution in every duplicate samples, mixture is centrifugal, and supernatant HPLC analyzes.
The absorption and transport situation of epimedium aglucone microemulsion is investigated in this test with the Caco-2 cell model:
When concentration is 20 μ M:
The AP side is 8.38 * 10 to the transport velocity of BL side
-4Cm/sec, the BL side is 6.88 * 10 to the transport velocity of AP side
-4Cm/sec.
When concentration is 80 μ M:
The AP side is 9.59 * 10 to the transport velocity of BL side
-4Cm/sec, the BL side is 9.04 * 10 to the transport velocity of AP side
-4Cm/sec.
As seen: in each concentration, absorption rate is greater than effluxing speed, and the epimedium aglucone microemulsion helps the absorption of epimedium aglucone.
Embodiment 21
Absorbing state in the body of rat intestine perfusion model research epimedium aglucone microemulsion
Anesthesia SD rat, place on the little electric blanket and light under, keep normal body temperature, 4cm opens the abdominal cavity along the ventrimeson otch, insert the bile conduit near the duodenum place, plug in polyethylene tube respectively at duodenum, jejunum, ileum and colon two ends, fix with surgical thread, the gauze that experiment Shi Yong etc. oozes the normal saline dipping is covered in the intestinal tissue surface to preserve moisture.Prepare the epimedium aglucone microemulsion solution as perfusate with HBSS solution.Change perfusate with waiting after oozing the normal saline flushing intestinal contents, with a constant speed pump perfusion enteric cavity.Collect perfusate in the outlet every 30min, the length of small intestinal is measured in the perfusion back.Outlet Chinese medicine concentration detects with HPLC.
The result: each intestinal segment of epimedium aglucone microemulsion absorbs good, and average infiltration coefficient is 3.87, and percent absorption is 72.96%.
Embodiment 22:
Differentiation assays icariin microemulsion anti-osteoporosis activity is cultivated and induced to bone marrow stroma stem cell
Alkali phosphatase (ALP) is osteoblastic significant enzyme, dyes with it usually and identifies osteoblast.Get the rat marrow stroma stem cell and cultivate, measure cell ALP enzymatic activity (CAKP dye liquor purchase build up in Nanjing bio-engineering research institute) with diazol method (improvement KaplowShi method) in cultivating the 12nd day.
Cell culture and administration:
Put to death Mus, 75% is alcohol-pickled, gets Mus lower limb (last two joints), with separation such as muscle.Open the Os Mus pulp cavity in Biohazard Safety Equipment, bone marrow stroma stem cell is blown out from one of medullary cavity with syringe, swap operation is to colourless.With syringe streak cell is blown and beaten into single cell suspension repeatedly to tube wall, visible DMEM becomes muddy by clarification.Add Vc in the cell culture medium, two anti-, dexamethasone etc. add medicine (epimedium aglucone microemulsion) simultaneously.Not add the medicine group as blank.Put into incubator (37%, 5%CO
2) cultivate.Cell changed liquid every three days.Carried out CAKP dyeing on the 12nd day in cultivating.
By Figure 21 a and b as seen: the icariin microemulsion group is for the blank group, and cell is seen many and deep dyed color, the ALP increased activity.
Claims (4)
1. icariin microemulsion, it is characterized in that: it is to be active component with the epimedium aglucone, it is dissolved in oil or the ester, add that the microemulsion particle diameter that emulsifying agent, co-emulsifier and water are prepared from is the microemulsion of 10～100nm, described oil is oleic acid, ethyl oleate, isopropyl myristate or Oleum Ricini
Wherein, the mass percent of each component is:
The saturated solution 0.66～6.02% of the oil of epimedium aglucone
Emulsifying agent 10～50%
Co-emulsifier 10～38.17%
Surplus is water, Hank ' s balanced salt solution, PBS buffer or normal saline.
2. microemulsion according to claim 1 is characterized in that: described emulsifying agent is OP.
3. be glycerol, isopropyl alcohol, Macrogol 200, Liquid Macrogol, PEG400 or 1 according to the described co-emulsifier of claim 1, the 2-propylene glycol.
4. a method for preparing the described icariin microemulsion of claim 1 is characterized in that it comprises the steps:
The preparation of step 1. epimedium aglucone oil solution: excessive epimedium aglucone is dissolved in the oil, mixes to make abundant dissolving, centrifugally remove undissolved epimedium aglucone, the saturated solution of oil of epimedium aglucone;
Step 2. takes by weighing the saturated solution of oil of the epimedium aglucone of emulsifying agent, co-emulsifier, water, step 1 gained by prescription;
Step 3. stirs four kinds of components that step 2 took by weighing and makes abundant mixing, gets the icariin microemulsion of clear.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105476957A (en) * | 2014-12-18 | 2016-04-13 | 北京珅奥基医药科技有限公司 | Icaritin injection and preparation method and application thereof |
| WO2016127925A1 (en) * | 2015-02-13 | 2016-08-18 | 北京盛诺基医药科技有限公司 | Drug combination of flavonoid compound and use of same |
| CN111481640A (en) * | 2020-04-27 | 2020-08-04 | 江苏省中医药研究院 | Anti-liver cancer microemulsion nano composition and application thereof |
-
2009
- 2009-03-27 CN CNA2009100299281A patent/CN101513388A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105476957A (en) * | 2014-12-18 | 2016-04-13 | 北京珅奥基医药科技有限公司 | Icaritin injection and preparation method and application thereof |
| CN105476957B (en) * | 2014-12-18 | 2021-04-20 | 北京珅奥基医药科技有限公司 | Acoradine injection and preparation method and application thereof |
| WO2016127925A1 (en) * | 2015-02-13 | 2016-08-18 | 北京盛诺基医药科技有限公司 | Drug combination of flavonoid compound and use of same |
| CN107205982A (en) * | 2015-02-13 | 2017-09-26 | 北京盛诺基医药科技有限公司 | A kind of medical composition and its use of chromocor compound |
| CN111481640A (en) * | 2020-04-27 | 2020-08-04 | 江苏省中医药研究院 | Anti-liver cancer microemulsion nano composition and application thereof |
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