CN105919973A

CN105919973A - Application of solid lipid nanoparticles as antidepressant drug carrier

Info

Publication number: CN105919973A
Application number: CN201610262638.1A
Authority: CN
Inventors: 汪世龙; 朱融融; 贺晓烈
Original assignee: Tongji University
Current assignee: Tongji University
Priority date: 2016-04-26
Filing date: 2016-04-26
Publication date: 2016-09-07

Abstract

The invention relates to an application of solid lipid nanoparticles as an antidepressant drug carrier. Specifically, an antidepressant drug is prepared by the following steps: preparing 0.1-0.2g of curcumin, 0.1-0.3g of folic acid, 0.5-0.15g of lecithin and 0.001-0.0003g of an HU-211 mixed chloroform solution, totaling 10-20ml; then, adding to 20-40ml of water which contains 0.2-0.3g of Myrj52, and stirring under 1000-1200rpm at 70-80 DEG C until the total volume of the mixed solution is 5-8ml left; adding 10-15ml of ice water, and continuing to stir under 1000-1200rpm at 0-4 DEG C for 2-3h; and centrifuging and discarding a supernatant, freeze-drying and preserving at 0-4 DEG C. The solid lipid nanoparticles, as the antidepressant drug carrier, can improve the water solubility and stability of the antidepressant drug, increase the bioavailability of the drug, control the release of the drug and improve a targeting property on treating depression.

Description

Solid lipid nanoparticle is as the application of antidepressant drug carrier

Technical field

The invention belongs to biomaterial and materia medica crossing domain, relate to the lipid nanoparticle application as antidepressant drug carrier.

Background technology

Depression is a kind of harm able-bodied common affective disorder disease of the whole mankind, with the notable and lasting low syndrome as principal character of mental state.The lifetime prevalence of depression is up to 15%, and the major depressive disorder of 15% can cause death because of suicide.

For the pathogenesis of depression, there is notable side effect and include cardiac toxicity in current medicine, sexual dysfunction and sleep disordered etc., and some antidepressant drug is difficult to through blood brain barrier.For solving problem above, present invention uses the hypotoxic curcumin with light spectrality and easily through the dexabinol (HU-211) of blood brain barrier.

Although curcumin has been shown to have antidepressant effect, but low and poor biocompatibility the shortcoming of water solublity limits its application, HU-211 there is also the problem of poor stability in biosystem equally, and the treatment that nano composite system is depression developed based on nano material provides a kind of new method.

Solid lipid nanoparticle (Solid lipid nanoparticles, SLN) it is the submicron drug-supplying system of new generation grown up early 1990s, refer to lipoid such as lecithin, the triacylglycerol etc. of solid natural or synthesis as carrier, by medicine parcel or clamp the solid micelle drug-supplying system made in lipoid core.SLN is widely used in the carrier as antitumor drug, as the carrier of transdermal delivery system, as the carrier of dosing eyes system, the carrier as peptide medicament and the carrier as sunscreen products.

Summary of the invention

It is an object of the invention to provide the application as antidepressant drug carrier of a kind of solid lipid nanoparticle.

The solid lipid nanoparticle that the present invention proposes is as the application of antidepressant drug carrier, and described antidepressant drug is primarily referred to as curcumin and HU-211, specifically comprises the following steps that

Preparation 0.1 ~ 0.2g curcumin, 0.1 ~ 0.3g folic acid, 0.5 ~ 0.15g lecithin and the mixing chloroformic solution totally 10 ~ 20ml of 0.001 ~ 0.0003g HU-211, it is subsequently adding in the 20 ~ 40ml water containing 0.2 ~ 0.3g Myrj52, and stir to mixed solution cumulative volume remaining as 5 ~ 8ml under 1000 ~ 1200rpm and 70 ~ 80 ° of C, add 10 ~ 15ml frozen water and continue to stir 2 ~ 3 hours under 1000 ~ 1200rpm and 0 ~ 4 ° of C.Being centrifuged and abandon supernatant, after lyophilization, 0 ~ 4 ° of C preserves.

In the present invention, it is 20 ~ 300nm that described solid lipid nanoparticle carries the mean particle diameter after antidepressant drug, and surface zeta potential current potential is-22.0 ~-21.3 mV.

Solid lipid nanoparticle is better than other nano material compound systems: (1) good biocompatibility；(2) drug release is controlled；(3) particle diameter is little makes its circulation time in vivo long；(4) drug loading is high；(5) biodegradable, bio-compatible；(6) hypotoxicity；(7) can pass through blood brain barrier.

To achieve these goals, the present invention utilizes low temperature emulsified method, carries curcumin and HU-211 compound system for Material synthesis SLN with curcumin, folic acid, lecithin, HU-211 and Myrj52, and obtains sample by lyophilization.

The beneficial effects of the present invention is: solid lipid nanoparticle bag is carried curcumin and HU-211 nanocomposite applications in treating depression, improve curcumin and the curative effect of HU-211 treatment depression, overcoming the application limitation of curcumin and HU-211, the clinical treatment for depression provides new method.

Accompanying drawing explanation

Fig. 1 is that SLN carries curcumin and HU-211 nano composite system transmission electron microscope picture.

Fig. 2 is that SLN carries curcumin and HU-211 nano composite system zeta potential diagram.

Fig. 3 is that SLN carries curcumin and the elution profiles of HU-211 nano composite system release medicine.

Fig. 4 is that SLN carries curcumin and the HU-211 MTT to depression model PC12 cell.

Fig. 5 is that SLN carries curcumin and HU-211 improves depression model mouse movement ability.

Fig. 6 is that SLN carries curcumin and HU-211 tissue distribution and targeting situation.

Detailed description of the invention

Embodiment 1

Preparation curcumin (0.1 ~ 0.2g), folic acid (0.1 ~ 0.3g), the mixing chloroformic solution totally 10 ~ 20ml of lecithin (0.5 ~ 0.15g) and HU-211 (0.001 ~ 0.0003g), it is subsequently adding in the 20 ~ 40ml water containing Myrj52 (0.2 ~ 0.3g) and stirs under 1000 ~ 1200rpm and 70 ~ 80 ° of C and about remain 5 ~ 8ml to mixed solution cumulative volume, add 10 ~ 15ml frozen water and continue to stir 2 ~ 3 hours under 1000 ~ 1200rpm and 0 ~ 4 ° of C.Being centrifuged and abandon supernatant, after lyophilization, 0 ~ 4 ° of C preserves.

Use 1 ~ 2% Sodium phosphotungstate as negative staining liquid, take a small amount of material solution and drop to be covered with on the copper mesh of carbon film, stand 5 ~ 10min, blot suspension with filter paper, then drip 1 ~ 2%(w/v) Sodium phosphotungstate negative staining 5 ~ 10min, observes form under JEOL type transmission electron microscope.Material Zeta current potential uses Malvern zeta potentiometer to measure.

Embodiment 2

2 ~ 5ml SLN carries curcumin and HU-211 Nanocomposite solution (10mg/ml) puts into a bag filter (blocking size 14,000 ~ 20,000 Da).Then bag filter put into 400 ~ 500ml PBS solution (pH 7.4) wherein comprise 8 ~ 10% Tween-80 (v/v) and under 100 ~ 2000rpm and 36 ~ 38 ° of C stir.In the specific time, taking out 4 ~ 5ml solution, by the content of UV spectrophotometer measuring curcumin, the identical fresh solution of same volume adds to keep cumulative volume constant.

Embodiment 3

Cell cultivation process adds corticosterone and obtains depression cell model, and PC12 cell is incubated at 96 orifice plates, adds medicine, arrange comparison after plating cells 24h.Packet transaction: corticosterone adds PBS, comparison, corticosterone adds fluoxetine, and corticosterone adds HU-211, and corticosterone adds Cur, and corticosterone adds Cur/SLNs, and corticosterone adds Cur/SLNs-HU-211.After continuing to cultivate 24h, every hole adds 20 μ l MTT solution (5mg/ml, i.e. 0.5%MTT), continues to cultivate 4h.Terminate cultivating, suck culture fluid in hole.Every hole adds 150 μ l dimethyl sulfoxide, low-speed oscillation 10 ~ 20min on shaking table, makes crystal fully dissolve.The light absorption value in each hole is measured at enzyme-linked immunosorbent assay instrument OD 490/560nm.

Embodiment 4

Successive administration (corticosterone) sets up mice depression model in 2 ~ 3 weeks.After administration processes 2 ~ 3 weeks the most respectively, use Behaviors survey to evaluate SLN and carry curcumin and HU-211 nano composite material to animal depression and improving situation.Packet transaction: PBS, normal control, fluoxetine, HU-211, Cur, Cur/SLNs, Cur/SLNs-HU-211.Turn-club test is the important indicator evaluating animal sports ability.1 ~ 2h after last administration, it is on 0 ~ 40 rev/min of transfer rod that each group of mice is placed on rotating speed, records the mice time of staying on transfer rod.Finally, take mice conscience spleen lung kidney brain analysis and measure drug entities distribution situation, evaluate the distribution of material and whether there is targeting.

Fig. 1 is that SLN carries curcumin and HU-211 nano composite system transmission electron microscope picture, and mean particle diameter is 20 ~ 300nm.

Fig. 2 is that SLN carries curcumin and HU-211 nano composite system zeta potential diagram, and surface zeta potential current potential is-22.0 ~-21.3 mV.

Fig. 3 is that SLN carries curcumin and the elution profiles of HU-211 nano composite system release medicine, and compound system can continue slowly to discharge curcumin in 7 days, the medicine of the most releasable about 77%.

Fig. 4 is that SLN carries curcumin and the HU-211 MTT to depression model PC12 cell, with normal group cell for comparison for 100%, individually corticosterone processes 24 hour cell survival rates is 50%, it is 93% that positive drug group fluoxetine processes cell survival rate, HU-211 group cell survival rate is 60%, curcumin group cell survival rate is 62%, and it is 65% that SLN carries curcumin group cell survival rate, and SLN carries curcumin and HU-211 group cell survival rate is 73%.SLN carries curcumin and HU-211 compound system and has the murder by poisoning from corticosterone of the more preferable protection PC12 cell compared to HU-211, curcumin and SLN lift-launch curcumin.

Fig. 5 is that SLN carries curcumin and HU-211 improves depression model mouse movement ability, each process group time of staying on transfer rod is respectively 247,300,300,247,242,270,293 seconds, SLN carried the mice after curcumin and HU-211 compound system carry curcumin process compared to HU-211, curcumin and SLN and has higher motor capacity.

Fig. 6 is that SLN carries curcumin and HU-211 tissue distribution and targeting situation, in brain, distribution is compared its hetero-organization and is wanted height, and after SLN lift-launch curcumin and HU-211 compound system carry curcumin process mice compared to curcumin and SLN, in brain, curcumin concentration wants height.

Claims

1. solid lipid nanoparticle is as the application of antidepressant drug carrier, it is characterised in that described antidepressant drug is primarily referred to as curcumin and HU-211, specifically comprises the following steps that

Preparation 0.1 ~ 0.2g curcumin, 0.1 ~ 0.3g folic acid, 0.5 ~ 0.15g lecithin and the mixing chloroformic solution totally 10 ~ 20ml of 0.001 ~ 0.0003g HU-211, it is subsequently adding in the 20 ~ 40ml water containing 0.2 ~ 0.3g Myrj52, and stir to mixed solution cumulative volume remaining as 5 ~ 8ml under 1000 ~ 1200rpm and 70 ~ 80 ° of C, add 10 ~ 15ml frozen water to continue to stir 2 ~ 3 hours under 1000 ~ 1200rpm and 0 ~ 4 ° of C, being centrifuged and abandon supernatant, after lyophilization, 0 ~ 4 ° of C preserves.

Application the most according to claim 1, it is characterised in that it is 20 ~ 300nm that described solid lipid nanoparticle carries the mean particle diameter after antidepressant drug, and surface zeta potential current potential is-22.0 ~-21.3 mV。