CN103494820B - Two medicine carrying PLGA nanoparticle of a kind of nimodipine/ligustrazine and preparation method thereof - Google Patents
Two medicine carrying PLGA nanoparticle of a kind of nimodipine/ligustrazine and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to the two medicine carrying PLGA nanoparticle of a kind of nimodipine/ligustrazine, obtain by the following method: precision takes nimodipine, ligustrazine phosphate and the Poly(D,L-lactide-co-glycolide that mass ratio is 1 ﹕ 5 ~ 20 ﹕ 40 ~ 60, be dissolved in acetone, as organic facies; The PVA aqueous solution of mass concentration 0.5 ~ 1.0% is as aqueous phase; Under agitation organic facies slowly instilled in aqueous phase, drip and finish, 40 ~ 50 DEG C of constant temperature continue stirring 2 ~ 4h, fling to organic solvent, centrifugal, and precipitation lyophilizing after distilled water wash 2 ~ 3 times, obtains the two medicine carrying PLGA nanoparticle of described nimodipine/ligustrazine.The present invention take PLGA as carrier material, P-gp inhibitor TMP and NMD use in conjunction have been prepared NMD/TMP-PLGA-NPs, the outer row's effect of P-gp is inhibit while short, the easy generation cytotoxicity of drug half-life etc. avoiding simply share existence is not enough, facilitate the distribution of NMD to tissue, compared with single drug-carrying nanometer particle, there is some superiority.
Description
(1) technical field
The present invention relates to two medicine carrying PLGA nanoparticle (NMD/TMP-PLGA-NPs) of a kind of nimodipine/ligustrazine and preparation method thereof.
(2) background technology
Nimodipine (Nimodipine, NMD) is a class dihydropyridine calcium ion antagonist, by blood flow treatment cerebrovascular disease in expansion brain small artery and raising brain.But NMD water solublity is poor, commercially available NMD injection, mainly through dissolve with ethanol medicine, has certain zest; The NMD oral formulations biological half-life of Clinical practice is short, and first-pass effect is strong, and bioavailability only has 5% ~ 13%; Meanwhile, NMD is as P-glycoprotein(P-gp) substrate, be subject to the upper P-gp of blood brain barrier (blood-brain barrier, BBB) and arrange function influence outward, cause concentration in its brain lower, limit its therapeutic effect when cerebral vascular attack.Ligustrazine (Tetramethylpyrazine, TMP) is a class Pyrazine alkaloid, and its pharmacological action is mainly reflected in and improves blood supply and protection ischemic tissue.Research shows, TMP can lower the expression of P-gp, the outer row of P-gp substrate in restrictive cell; And the atpase activity of P-gp can be suppressed, thus suppress the function of P-gp, increase blood-brain barrier permeability, be conducive to medicine and enter cerebral tissue.Therefore TMP and NMD is share, be expected to improve concentration in NMD brain, but NMD and TMP simply share that still to there is drug half-life short, must the problem of frequent drug administration, and TMP excessive concentration may cause NMD excessive concentration in brain in short-term, toxicity is produced to normal cell or tissue.
(3) summary of the invention
The present invention seeks to, for solving the deficiency existed when NMD and TMP simply share in prior art, to provide two medicine carrying PLGA nanoparticle of a kind of nimodipine/ligustrazine and preparation method thereof.
The two medicine carrying PLGA nanoparticle of a kind of nimodipine/ligustrazine, obtain by the following method: precision takes nimodipine (NMD), ligustrazine phosphate (TMP) and the Poly(D,L-lactide-co-glycolide (PLGA) that mass ratio is 1:5 ~ 20:40 ~ 60, be dissolved in acetone, as organic facies; The PVA aqueous solution of mass concentration 0.5 ~ 1.0% is as aqueous phase; Under agitation organic facies slowly instilled in aqueous phase, drip and finish, 40 ~ 50 DEG C of constant temperature continue stirring 2 ~ 4h, fling to organic solvent, centrifugal, and precipitation lyophilizing after distilled water wash 2 ~ 3 times, obtains the two medicine carrying PLGA nanoparticle of described nimodipine/ligustrazine.Described PLGA molecular weight, between 10000 ~ 50000Da, is preferably 20000Da, and described PVA molecular weight, between 10000 ~ 50000Da, is preferably 13000 ~ 23000Da.
Preferably, described nimodipine, ligustrazine phosphate and poly lactic-co-glycolic acid mass ratio are 1:10:50.
Nanoparticle (nanoparticles, NPs) is a kind of new drug carrier, has slow releasing medicine, reduces the advantages such as toxic and side effects.NMD and TMP wraps and is loaded in NPs and can delays drug release by the present invention jointly, avoids the strong first pass effect of hepar of NMD, reduces because of the excessive cell or tissue toxicity caused of TMP concentration in short-term, overcomes its deficiency while effectively playing drug combination advantage.
The invention still further relates to the method for the two medicine carrying PLGA nanoparticle of nimodipine/ligustrazine described in preparation, described method comprises: precision takes nimodipine, ligustrazine phosphate and the Poly(D,L-lactide-co-glycolide that mass ratio is 1:5 ~ 20:40 ~ 60, be dissolved in proper amount of acetone, as organic facies; The PVA aqueous solution of mass concentration 0.5 ~ 1.0% is as aqueous phase; At stirring (500 ~ 800rmin
-1) under organic facies is slowly instilled in aqueous phase, drip and finish, 40 ~ 50 DEG C of constant temperature continue stirring 2 ~ 4h(500 ~ 800rmin
-1), fling to organic solvent, 10000 ~ 15000rmin
-1centrifugal 20 ~ 30min, gets lyophilizing after precipitation distilled water wash 2 ~ 3 times, obtains the two medicine carrying PLGA nanoparticle of described nimodipine/ligustrazine.
Preferably, the ratio of described nimodipine, ligustrazine phosphate and poly lactic-co-glycolic acid consumption is 1:10:50.
Preferably, described acetone consumption is 2 ~ 10mL/mg nimodipine, and aqueous phase volume is 5 ~ 20 times of organic facies volume.
Beneficial effect of the present invention is mainly reflected in: the present invention take PLGA as carrier material, P-gp inhibitor TMP and NMD use in conjunction have been prepared NMD/TMP-PLGA-NPs, the outer row's effect of P-gp is inhibit while short, the easy generation cytotoxicity of drug half-life etc. avoiding simply share existence is not enough, facilitate the distribution of NMD to tissue, compared with single drug-carrying nanometer particle, there is some superiority.
(4) accompanying drawing explanation
Fig. 1 is NMD/TMP-PLGA-NPs transmission electron microscope photo (× 60000);
Fig. 2 is NMD/TMP-PLGA-NPs particle size distribution;
Fig. 3 is NMD/TMP-PLGA-NPs Zeta potential;
Fig. 4 is NMD, TMP tablets in vitro curve (n=3);
Fig. 5 is blank rat plasma (A), and blank rat plasma adds NMD standard solution (B), rat plasma sample (C) high-efficient liquid phase chromatogram after tail intravenously administrable NMD/TMP-PLGA-NPs;
Fig. 6 is Drug-time curve after rat tail vein administration NMD suspension, NMD/TMP suspension, NMD-PLGA-NPs and NMD/TMP-PLGA-NPs.
(5) detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further, but protection scope of the present invention is not limited in this:
Embodiment 1:
1. instrument and material
1.1 instrument
UV-1700 ultraviolet spectrophotometer (Japanese Shimadzu Corporation); Agilent1200 high performance liquid chromatograph (Anjelen Sci. & Tech. Inc of the U.S.); 380ZLS laser granulometry (Nico mp company of the U.S.); JEM-1200EX transmission electron microscope (Japanese JEOL company); Labconc o freezer dryer (Labconco company of the U.S.); Mill-Q Superpure water machine (Millpore company of the U.S.); KQ5200DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instruments Co., Ltd.); TGL-16B high speed tabletop centrifuge (Anting Scientific Instrument Factory, Shanghai); CP225D electronic balance (Beijing Sai Duolisi scientific instrument company limited); Nitrogen purges instrument (Hangzhou Ao Sheng Instrument Ltd.); Bag filter (upper sea green bird development in science and technology company limited, Mw=14000Da); HZ-9212S water-bath constant temperature oscillator (Taicang, Jiangsu Hua Lida experimental facilities company).
1.2 medicine and reagent
Nimodipine (Hubei Heng Shuo pharmaceutcal corporation, Ltd, purity >98%, lot number 20100302); Nimodipine reference substance (Chinese food drug assay institute, lot number 100270-200002); Ligustrazine phosphate (Fei Yu bio tech ltd, Nantong, purity >95%, lot number 20110215); Ligustrazine phosphate reference substance (Chinese food drug assay institute, lot number 110817-201008); Poly(D,L-lactide-co-glycolide (Jinan Dai Gang biological engineering company limited, Mw=20000Da); Polyvinyl alcohol (Sigma Co., USA, Mw=13000 ~ 23000Da, lot number 10918AE); Methanol (chromatographically pure, Honeywell Burdick & Jackson company of the U.S., lot number 10071753); Other reagent are domestic analytical pure.
1.3 laboratory animal
Cleaning grade SD rat, male and female dual-purpose, body weight (220 ± 10) g(Zhejiang University of Traditional Chinese Medicine Experimental Animal Center, (quality certification number: SCXK Zhejiang 2008-0036)); All zooperies are all carried out according to Zhejiang University's animal feeding and guide for use.
2. method
The preparation of 2.1 nanoparticles
The emulsified solvent sedimentation method are adopted to prepare nanoparticle: precision takes 1.0mg NMD, 10mg TMP and 50mg PLGA is dissolved in 3mL acetone, as organic facies; 0.75%PVA aqueous solution 36mL is as aqueous phase.At 800rmin
-1organic facies slowly instilled in aqueous phase with No. 5 syringe needles under mixing speed, drip and finish, 40 DEG C of constant temperature continue 800rmin
-1stir 3h, obtain NMD/TMP-PLGA-NPs suspension, fling to organic solvent, ultracentrifugation (13000rmin
-1) 30min, after precipitation distilled water wash 3 times, namely lyophilizing obtains NMD/TMP-PLGA-NPs.
NMD-PLGA-NPs(nimodipine medicine carrying PLGA nanoparticle) preparation method all same NMD/TMP-PLGA-NPs except not adding TMP.
2.2 forms, particle diameter and Zeta potential
Get appropriate NMD/TMP-PLGA-NPs lyophilized powder, add water and redissolve for suspendible drop is on copper mesh, blot with filter paper after leaving standstill 10min, then drip mass fraction 2.0% Salkowski's solution negative staining 3min on copper mesh, naturally volatilize, by transmission electron microscope observation form and photographs; Get NMD/TMP-PLGA-NPs suspension, after appropriate distilled water diluting, adopt laser granulometry to measure mean diameter, particle size distribution and Zeta potential.
The mensuration of 2.3 envelop rates and drug loading
2.3.1 chromatographic condition
NMD chromatographic condition chromatographic column: Thermo Hypersil Gold C
18post (250mm × 4.6mm, 5 μm); Mobile phase: methanol-water (70:30); Determined wavelength: 238nm; Flow velocity: 1.0mLmin
-1; Column temperature: 35 DEG C; Sampling volume: 20 μ L.
TMP chromatographic condition chromatographic column: Thermo Hypersil Gold C
18post (250mm × 4.6mm, 5 μm); Mobile phase: methanol-water (45:55); Determined wavelength: 280nm; Flow velocity: 1.0mLmin
-1; Column temperature: 35 DEG C; Sampling volume: 20 μ L.
2.3.2 linear relationship is investigated
NMD linear relationship investigates that accurate to take drying under reduced pressure appropriate to the NMD reference substance of constant-quality, put in 10mL volumetric flask, and be diluted to scale with dissolve with methanol, obtaining mass concentration is 0.196mgmL
-1nMD reference substance storing solution.Precision measures storing solution in right amount in 10mL volumetric flask respectively, with methanol dilution to scale, obtains concentration and is respectively 1.96,3.92,9.80,19.60,39.20,58.80 μ gmL
-1control series product solution, measure by NMD chromatographic condition under " 2.3.1 " item, with integrating peak areas value (A), linear regression carried out to mass concentration (C), obtain regression equation A=146.10C+13.23(r=0.9997), show that NMD is at 1.96 ~ 58.80 μ gmL
-1interior peak area and drug level are good linear relationship.
TMP linear relationship investigates that accurate to take drying under reduced pressure appropriate to the TMP reference substance of constant-quality, put in 10mL volumetric flask, and be diluted to scale with dissolve with methanol, obtaining mass concentration is 0.196mgmL
-1tMP reference substance storing solution.Precision measures storing solution in right amount in 10mL volumetric flask respectively, and with methanol dilution to scale, obtaining concentration is 1.25,2.50,5.00,10.00,12.50,23.00 μ gmL
-1control series product solution, measure by TMP chromatographic condition under " 2.3.1 " item, with integrating peak areas value (A), linear regression carried out to mass concentration (C), obtain regression equation A=42.587C-13.869(r=0.9999), TMP is at 1.25 ~ 46.00 μ gmL
-1interior peak area and drug level are good linear relationship.
2.3.3 envelop rate and drug loading measure precision and measure NMD/TMP-PLGA-NPs suspension 1mL, are placed in tool plug centrifuge tube, 13000rmin under room temperature
-1centrifugal 30min, gets supernatant 200 μ L in 10mL volumetric flask, by dilution in acetonitrile to scale, filters, get subsequent filtrate and measure free NMD and TMP content respectively by chromatographic condition under " 2.3.1 " item through 0.22 μm of microporous filter membrane; Separately get NMD/TMP-PLGA-NPs suspension 200 μ L in 10mL volumetric flask, be diluted to scale with appropriate acetonitrile breakdown of emulsion, filter through 0.22 μm of microporous filter membrane, subsequent filtrate measures NMD and TMP content total in NMD/TMP-PLGA-NPs respectively by chromatographic condition under " 2.3.1 " item; Envelop rate (EE%) and the drug loading (DL%) of NMD and TMP in NPs is calculated respectively by following formula.
EE%＝(W
0–W
1)/W
0×100%
DL%＝(W
0–W
1)/W
t×100%
Wherein, W
0for the total medication amount in NPs; W
1for the free drug amount in NPs; W
t: the gross weight of NPs.
2.4 tablets in vitro researchs
The PBS solution chosen containing volume fraction 30% ethanol is release medium (pH7.4), investigates the tablets in vitro behavior of NMD/TMP-PLGA-NPs.Precision takes each 3 parts of appropriate NMD/TMP former medicine mixed-powder, NMD/TMP-PLGA-NPs lyophilized powder respectively, be scattered in 2mL release medium, be placed in the bag filter (Mw=14000Da) of anticipating, seal after getting rid of bubble, be placed in 200mL release medium, in (37 ± 0.5) DEG C water bath with thermostatic control vibration (75rmin
-1), respectively at 0.1,0.25,0.5,0.75,1,1.25,1.5,1.75,2,2.5,3,4,5,6,8,12,24h accurately samples 1mL, and add equivalent equality of temperature fresh dissolution medium immediately, sample filters through 0.22 μm of microporous filter membrane, get subsequent filtrate and measure release medium drug content by chromatographic condition under " 2.3.1 " item, calculate cumulative release rate (Q%).
2.5NMD/TMP-PLGA-NPs rat Internal pharmacokinetics is studied
2.5.1 plasma sample process precision pipettes plasma sample 200 μ L and puts in 2mL tool plug centrifuge tube, and add acetonitrile 800 μ L, vortex mixed 3min, in 10000rmin
-1centrifugal 10min, gets supernatant and dries up by nitrogen current in 40 DEG C, and residue 200 μ L dissolve with methanol, in 10000rmin after abundant vortex oscillation
-1centrifugal 10min, gets the analysis of supernatant 20 μ L sample introduction.
2.5.2 the foundation of NMD assay method in blood plasma
Chromatographic condition chromatographic column: Thermo Hypersil Gold C
18post (250mm × 4.6mm, 5 μm); Mobile phase: methanol-water (70:30); Flow velocity: 1.0mLmin
-1; Column temperature: 30 DEG C; Determined wavelength 238nm.Sampling volume: 20 μ L.
Method specificity is investigated and is taken 1mg NMD, 10mg TMP in 10mL volumetric flask, is diluted to scale, obtains NMD/TMP reference substance solution with dissolve with methanol.Get rat blank plasma respectively, rat blank plasma adds NMD/TMP reference substance solution and rat plasma sample after rat tail vein injection NMD/TMP-PLGA-NPs, measure by sample introduction after method process under " 2.5.1 " item.
It is appropriate to the NMD reference substance of constant weight that the preparation precision of standard curve takes drying under reduced pressure, and be placed in 25mL measuring bottle, add dissolve with methanol and be diluted to scale, being made into mass concentration is 0.040 μ gL
-1reference substance solution, be diluted to series concentration before use.
Accurate absorption blank plasma 1mL7 part, adds the reference substance solution 20 μ L of series concentration respectively, vortex mixed, obtains mass concentration and be respectively 6.25,12.50,25.00,50.00,100.00,200.00,400.00 μ gL
-1nMD Serial plasma standard solution, measure by sample introduction after method process under " 2.5.1 " item, carry out linear regression, Criterion curvilinear equation with the concentration (C) of peak area (A) to NMD in blood plasma.
The method response rate and precision measure precision and measure NMD reference substance solution in right amount, are mixed with 50.00,100.00,200.00 μ gL
-1the plasma sample of basic, normal, high 3 mass concentrations, measure by sample introduction after method process under " 2.5.1 " item, each concentration parallel assay 3 times, calculate the response rate.In a few days measuring 5 times respectively, METHOD FOR CONTINUOUS DETERMINATION 5d(every day 1 time), calculate in a few days and day to day precision.2.5.3 dosage regimen and blood specimen collection
Take 1mg NMD and NMD/TMP(NMD1mg, TMP10mg) former medicated powder end in 10mL volumetric flask, use ethanol in proper amount ultrasonic dissolution, with 0.9% normal saline dilution to scale, obtain NMD suspension and NMD/TMP suspension.Take 3.6g NMD-PLGA-NPs and NMD/TMP-PLGA-NPs lyophilized powder in 10mL volumetric flask, be diluted to scale with appropriate 0.9% normal saline ultrasonic dissolution, obtain NMD-PLGA-NPs and NMD/TMP-PLGA-NPs suspension.
Get healthy SD rat 32, fasting 12h, freely drinks water, and is divided into 4 groups at random, often organizes 8.By NMD2mgkg
-1single dose tail vein injection gives NMD suspension, NMD/TMP suspension, NMD-PLGA-NPs suspension and NMD/TMP-PLGA-NPs suspension respectively, respectively at 2,5,15,30,60,90,120,180,300 after administration, 420min gets blood 0.5mL through femoral artery, put in heparin sodium pretreated 2mL tool plug centrifuge tube, 3000rmin
-1centrifugal 10min, after separated plasma, puts-80 DEG C of cryogenic refrigerators and preserves to be measured.Give 0.9% normal saline of equivalent after getting blood immediately with microsyringe at every turn.
3 results
The character of 3.1NMD/TMP-PLGA-NPs
Observing NMD/TMP-PLGA-NPs under transmission electron microscope is similar round, size and distribution uniform, has no adhesion and gathering (Fig. 1) between particle.Particle size distribution result shows, and mean diameter is (631.6 ± 3.2) nm, and polydispersity coefficient is (0.097 ± 0.007) (Fig. 2); Zeta potential is (-19.25 ± 1.87) mV(Fig. 3).Recording NMD envelop rate in NMD/TMP-PLGA-NPs is (76.25 ± 1.18) %, and drug loading is (1.24 ± 0.01) %; TMP envelop rate is (39.30 ± 1.00) %, and drug loading is (6.34 ± 0.11) %.
3.2NMD/TMP-PLGA-NPs tablets in vitro
In pH7.4PBS medium (containing volume fraction 30% ethanol), NMD and TMP tablets in vitro curve is shown in Fig. 4.NMD/TMP-Sol(former medicine mixed-powder) in the release in release medium of two medicines all very fast, 0.75h NMD and TMP cumulative release percentage rate are respectively 57.28% and 56.67%, during 3h, two medicines discharge completely all substantially, and cumulative release percentage rate is respectively 95.5% and 95.39%.NMD/TMP-PLGA-NPs is respectively 27.10% and 23.61% at 0.75h NMD and TMP cumulative release percentage rate, compares, have obvious sustained releasing character with NMD/TMP-Sol; 24hNMD and TMP cumulative release percentage rate is respectively 75.6% and 55.3%.
3.4 pharmacokinetic studies
3.4.1 after method specificity blank rat plasma, blank rat plasma add NMD reference substance and rat tail vein drug administration by injection NMD/TMP-PLGA-NPs, plasma sample HPLC chromatogram is shown in Fig. 5.As shown in Figure 5, the method good separating effect set up, specificity is strong, impurity or endogenous material all not interference measurements in blood plasma.
3.4.2 in standard curve and methodological study result blood plasma, the standard curve equation of NMD is A=0.251C-0.046(r=0.9995), show that NMD plasma purity concentration is at 6.25 ~ 400.00 μ gL
-1internal linear relation relation is good.
3.4.3 the method response rate of precision, the basic, normal, high 3 kinds of plasma concentration NMD of method response rate investigation is respectively (90.08 ± 5.09) %, (93.96 ± 2.93) %, (90.55 ± 2.49); In a few days RSD is respectively 5.66%, 3.12%, 2.75%; In the daytime RSD is respectively 7.55%, 3.74%, 3.04%, meets methodology requirement.
3.4.4, after blood concentration-time curve and pharmacokinetic parameters rat tail vein inject NMD suspension, NMD/TMP suspension, NMD-PLGA-NPs suspension and NMD/TMP-PLGA-NPs suspension, mean blood plasma concentration-time graph is shown in Fig. 6.Data meet open two-compartment model after matching, and gained main pharmacokinetic parameters in table 1, and carries out statistical analysis to it.As shown in Table 1, NPs group (NMD-PLGA-NPs suspension and NMD/TMP-PLGA-NPs suspension) is compared, t with suspension group (NMD suspension and NMD/TMP suspension)
1/2significant prolongation (p<0.01), AUC significantly improves (p<0.01), illustrates that NPs has delayed the elimination of medicine; Two medicine coupling groups (NMD/TMP-Sol with NMD/TMP-PLGA-NPs) are compared with alone NMD group (NMD-Sol with NMD-PLGA-NPs), and CL value significantly improves (p<0.05), V
βvalue enlarges markedly (p<0.01), illustrates that TMP have impact on the behavior of NMD Internal pharmacokinetics to a certain extent.
Table 1: pharmacokinetic parameters after rat tail vein administration
*p<0.05,
**p<0.01,NMD-Sol vs NMD/TMP-Sol
△p<0.05,
△△p<0.01,NMD-PLGA-NPs vs NMD/TMP-PLGA-NPs
▲p<0.05,
▲▲p<0.01,NMD-Sol vs NMD-PLGA-NPs or NMD/TMP-PLGA-NPs
4 discuss
Two drug-carrying nanometer particle, as the new method of a kind of compatibility administration, efficacy enhancing and toxicity reducing, obtains concern in recent years gradually.The present invention take PLGA as carrier material, P-gp inhibitor TMP and NMD use in conjunction have been prepared NMD/TMP-PLGA-NPs, the outer row's effect of P-gp is inhibit while short, the easy generation cytotoxicity of drug half-life etc. avoiding simply share existence is not enough, facilitate the distribution of NMD to tissue, compared with single drug-carrying nanometer particle, there is some superiority.
The present invention adopts the emulsified solvent sedimentation method to prepare NMD/TMP-PLGA-NPs.Because TMP is soluble in water, easily depart from from oil phase in emulsion process and constantly diffuse in aqueous phase, envelop rate being reduced, and duct can be left on NPs surface, make its rate of release in elementary drug release process very fast.Therefore, be improve TMP envelop rate in NPs, the impact of each factor on envelop rate of this effects.Result shows, TMP envelop rate mainly affects by PLGA molecular weight and concentration, outer aqueous phase PVA concentration: increase with PLGA molecular weight and concentration, and organic facies viscosity increases, and TMP slows down to aqueous phase diffusion rate, with PLGA interaction of molecules time lengthening, envelop rate improves; Increase with aqueous phase PVA concentration, organic facies and aqueous phase interface surface tension reduce, more easily form less emulsion droplet, NPs particle diameter is reduced, aqueous phase viscosity increases simultaneously, be unfavorable for the diffusion of medicine to aqueous phase, therefore envelop rate improves, but too high PLGA molecular weight and concentration, PVA concentration can cause its particle diameter larger.Through investigating, this experiment employing molecular weight is the PLGA of 20000Da is carrier material (consumption is 50mg), and 0.75%PVA is that aqueous phase is to obtain better particle diameter and higher TMP envelop rate.
The dissolubility of NMD in water is minimum, when selecting common release medium, only has trace medicine in dissolution fluid.The equilbrium solubility of NMD in pH7.4PBS buffer solution (containing volume fraction 30% ethanol) is about 82.98 μ gmL
-1, the drug level after NPs discharges completely is 1.32 μ gmL
-1, meet sink conditions, therefore this experimental selection pH7.4PBS buffer solution (containing volume fraction 30% ethanol) is as vitro Release Medium.Result shows, and the NPs of this experiment preparation has certain slow release characteristic.
Pharmacokinetic studies result shows: compared with suspension group, NPs group t
1/2 βsignificant prolongation (p<0.01), CL significantly reduces (p<0.01), and AUC about improves 4 times, and prompting NPs has certain slow releasing function, and reduces the supersession rate of NMD; Compared with alone NMD group, two medicine coupling group t
1/2 αremarkable reduction (p<0.01), V
βsignificantly improve (p<0.05) with CL, AUC obviously reduces (p<0.05), and this conforms to P-gp inhibitor coupling result of study with other drug.T
1/2 αreduce prompting TMP and may facilitate the distribution of NMD to tissue to a certain extent, reduce medicine holdup time in blood plasma, effectively played the effect of P-gp inhibitor.
Claims (3)
1. prepare the method for the two medicine carrying PLGA nanoparticle of nimodipine/ligustrazine for one kind, described method comprises: precision takes nimodipine, ligustrazine phosphate and the Poly(D,L-lactide-co-glycolide that mass ratio is 1:5 ~ 20:40 ~ 60, be dissolved in acetone, as organic facies; The PVA aqueous solution of mass concentration 0.5 ~ 1.0% is as aqueous phase; Under agitation organic facies slowly instilled in aqueous phase, drip and finish, 40 ~ 50 DEG C of constant temperature continue stirring 2 ~ 4h, fling to organic solvent, 10000 ~ 15000rmin
-1centrifugal 20 ~ 30min, gets lyophilizing after precipitation distilled water wash 2 ~ 3 times, obtains the two medicine carrying PLGA nanoparticle of described nimodipine/ligustrazine.
2. the method for claim 1, is characterized in that the ratio of described nimodipine, ligustrazine phosphate and poly lactic-co-glycolic acid consumption is 1:10:50.
3. the method for claim 1, is characterized in that described acetone consumption is 2 ~ 10mL/mg nimodipine, and aqueous phase volume is 5 ~ 20 times of organic facies volume.
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Non-Patent Citations (3)
| Title |
|---|
| 川芎嗪联合尼莫地平治疗急性脑血管痉挛疗效观察;胡艳萍等;《现代中西医结合杂志》;20120331;第21卷(第7期);718－719 * |
| 川芎嗪聚乳酸纳米粒的制备和质量研究;曾莉等;《中成药》;20130228;第35卷(第2期);261－263 * |
| 范盈等.包载尼莫地平的聚乳酸-聚乙二醇嵌段共聚物纳米粒的制备和质量评价.《中国基层医药》.2007,第14卷(第11期),1767-1768. * |
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| CN103494820A (en) | 2014-01-08 |
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