CN104367549B - Contain psoralen adriamycin composite nanostructure lipid vector preparation and preparation method thereof - Google Patents
Contain psoralen adriamycin composite nanostructure lipid vector preparation and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to contain psoralen adriamycin composite nanostructure lipid vector preparation and preparation method thereof.The technical scheme of use is：It is made by weight ratio including following raw material：1-5 part of psoralen, 1-5 part of adriamycin, 40-120 parts of solid lipid, 10-30 parts of liquid fatty, 35-350 parts of fat-soluble emulsifier, 25-250 parts of water soluble emulsifier, 100-600 parts of surfactant.Two medicine preparations into composite nanostructure lipid carrier, are optimized and investigated to its formulation and technology and physicochemical property, prepare preferably nanostructured carrier by the present invention, and the multidrug resistance effect for reversing leukaemia for further research provides basis.
Description
Technical field
The invention belongs to pharmaceutics of Chinese drugs field, and in particular to a kind of composite Nano knot for containing psoralen and adriamycin
Structure lipid carrier and preparation method thereof.
Background technology
Tumour is one of major affliction of current harm human health.Invasion and metastasis of tumor is that malignant tumour is most common
A kind of biological behaviour and most essential feature, be also influence tumor patient existence and more after key factor.Therefore it is antitumor
Medicine is also the problem that those skilled in the art constantly researches and develops.
Psoralen (PSO) is the active ingredient extracted from legume, and fat-soluble effect is stronger, is a kind of calcium ion
Channel blocker, while can suppress pumping out for P-gp albumen, assists the effect of chemotherapeutics reversing multiple medicine resistance of tumor cells.Ah
Mycin (DOX) is anthracene ring antitumor medicinal, and its hydrochloride has water solubility, is mainly used in treatment leukaemia, lymph cancer and breast
Gland cancer etc., antitumor spectra is wide, but easily produces drug-resistant effect.At present, psoralen and adriamycin act solely on clinic
The research of leukaemia, but psoralen poorly water-soluble are treated, bioavilability is low, and doxorubicin hydrochloride water solubility effect is strong, but
Exocytosis is big, is also easy to produce resistance.Therefore need to seek a kind of effective preparation, to increase the dissolubility of psoralen, carry
High bioavilability, while reducing the exocytosis of adriamycin, strengthens antitumous effect, reduces toxic side effect.
Nano structured lipid carrier (NLC) is the novel Drug Delivery Systems developed on the basis of solid lipid nano granule (SLN),
Liquid fatty is added on the basis of solid lipid is carrier, the perfect crystal formation after Solid lipid solidification is upset, less medicine
Leakage.
The content of the invention
It is an object of the invention to by two medicine preparations into composite nanostructure lipid carrier, to its formulation and technology and reason
Change property to optimize and investigate, prepare preferably nanostructured carrier, be that further research reverses leukaemia's many
Medicine drug-resistant effect provides basis.
The technical solution adopted by the present invention is：One kind contains psoralen-adriamycin composite nanostructure lipid carrier system
Agent, including following raw material are made by weight ratio：
The solid lipid is stearic acid, cholesterol, palmitic acid, behenic acid, glycerin monostearate (GMS), glycerine palm fibre
Palmitic acid acid stearate Precirol ATO5), bi-tristearin, myristin, monopalmitin, behenic acid
One or two or more kinds of combinations of glyceride or glyceryl laurate ester.
The liquid fatty is Labraso, Miglyol 812N (LABRATAC
LIPOPHIE WL1349), isopropyl palmitate, isopropyl myristate, atoleine, soybean oil or one kind of oleic acid or two
Plant the mixing of the above.
The fat-soluble emulsifier is soybean lecithin or synthetic phospholipid.
The water soluble emulsifier be poloxamer, polyoxyethylene aliphatic alcohol ether, polyoxyethylene fatty acid ester, sodium taurocholate or
One or two or more kinds of mixing of NaTDC.
The poloxamer is poloxamer188, Pluronic/Lutrol F 108 or PLURONICS F87；It is preferred that PLURONICS F87.
The surfactant is Tween 80, polysorbate60 or polysorbas20；It is preferred that Tween 80.
The above-mentioned preparation method for containing psoralen-adriamycin composite nanostructure lipid vector preparation is as follows：
1) psoralen, solid lipid, liquid fatty and fat-soluble emulsifier are sequentially added in appropriate organic solvent, 70
Dissolved by heating in~80 DEG C of water-baths, ultrasonic (200~400W, 2~5min) is scattered to be mixed, and is used as organic phase；
The organic solvent is absolute ethyl alcohol；
2) adriamycin and water soluble emulsifier are dissolved in appropriate water for injection, dissolve by heating, stir in 70~80 DEG C of water-baths
Mix uniform, be used as interior aqueous phase；
3) surfactant is dissolved in appropriate water for injection, dissolves by heating, stir in 70~80 DEG C of water-baths, as
Outer aqueous phase；
4) under 800~1000r/min stirrings, interior aqueous phase is slowly injected into organic phase, 5~15min of stirring is formed just
Breast；
5) under 800~1000r/min stirrings, colostrum is slowly injected into outer aqueous phase, stirring and emulsifying formation emulsion；
6) poured into after emulsion is concentrated in frozen water rapidly, 0.5~1.5h of low-temperature setting, filtering with microporous membrane obtains liquid phase bag
Carry psoralen-adriamycin composite nanostructure lipid vector preparation；
The frozen water is 0~4 DEG C of water for injection；
It is preferred that, the micro porous filtration is to use aperture for 0.22 μm or 0.45 μm of membrane filtration；
7) to liquid phase contain in psoralen-adriamycin composite nanostructure lipid vector preparation add freeze drying protectant it is molten
Liquid, freeze-drying, obtains solid phase and contains psoralen-adriamycin composite nanostructure lipid vector preparation.
It is preferred that, the mass percent concentration of the frozen-dried protective agent solution is 2~10%；The freeze drying protectant is Portugal
Grape sugar, mannitol, sucrose, lactose, fructose, sorbierite, trehalose, maltose, dextran or one kind in amino acid or two
Plant the mixing of the above.
The beneficial effects of the invention are as follows：The present invention, using nano structured lipid carrier (NLC) by psoralen and adriamycin
Wrapped up, a kind of novel nano drug delivery system is made, the liquid fatty immiscible with Solid lipid is added to solid lipid
It is middle to form incomplete crystal formation, effectively prevent the leakage of medicine.Psoralen-Ah mould is contained by prepared by the method for the present invention
Plain composite nanostructure lipid vector preparation (PSO-DOX-NLC) is circular, similar round entity ball, and particle diameter is (128.7 ± 1.8)
Nm, polydispersity coefficient is 0.22 ± 0.01, and Zeta potential is (20.17 ± 0.31) mV, PSO envelop rates up to more than 70%, DOX
Envelop rate is up to more than 80%, and release in vitro meets Bidirectional power model, PSO and DOX drug solutions discharge substantially in 12h
Completely, in PSO-DOX-NLC nano-solutions there is burst effect in PSO in preceding 8h, and DOX is then sustained preferably, therefore the present invention
Containing psoralen-adriamycin composite nanostructure lipid vector preparation has good sustained release performance.The present invention, by that will mend
Bone fat element and adriamycin can improve bioavilability by being prepared into NLC, increase antitumous effect.
The present invention by by psoralen and adriamycin using composite nanostructure lipid carrier is prepared into, it is water-soluble
Adriamycin play therapeutic action, fat-soluble psoralen simultaneously play treatment and reverse leukaemia drug-resistant effect,
So as to effectively improve the aggregation of medicine in the cell, increase therapeutic effect, simultaneously because the advantage of its formulation, can slow down medicine
Excretion, reach slow releasing function.The present invention using simple preparation technology by both carry out be combined preparation, and obtain 70% with
On envelop rate, particle diameter is in 100nm or so, and technique is simple, is easy to industrialized production.
Brief description of the drawings
Figure 1A is the chromatogram of PSO solution in embodiment 1.
Figure 1B is the chromatogram of DOX solution in embodiment 1.
Fig. 1 C are the chromatograms of PSO-DOX mixed solutions in embodiment 1.
Fig. 1 D are the chromatograms of blank NLC in embodiment 1.
Fig. 1 E are the chromatograms of PSO-DOX-NLC solution in embodiment 1.
Fig. 2 is PSO-DOX-NLC prepared by embodiment 1 projection electron microscope.
Fig. 3 is PSO-DOX-NLC prepared by embodiment 1 grain size distribution.
Fig. 4 is PSO-DOX-NLC prepared by embodiment 1 Zeta potential distribution map.
Fig. 5 A are DOX and PSO-DOX-NLC drug release patterns in vitro in embodiment 1.
Fig. 5 B are PSO and PSO-DOX-NLC drug release patterns in vitro in embodiment 1.
Embodiment
Embodiment 1 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
(1) preparation method is as follows：
1) by 3mg psoralens (PSO), 104mg solid lipids (weight ratio, GMS：Precirol ATO5=1:1)、
26mg liquid fatties (Miglyol 812N (LABRATAC LIPOPHIE WL1349)) and 250mg fat-soluble emulsifiers
(soybean lecithin) is sequentially added in 20ml absolute ethyl alcohols, is dissolved by heating in 78 DEG C of water-baths, and ultrasonic disperse is mixed, as organic
Phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (PLURONICS F87) are dissolved in 1ml waters for injection, 78
Dissolved by heating in DEG C water-bath, stir, be used as interior aqueous phase；
3) 520mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under 1000r/min stirrings, above-mentioned colostrum is slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to about 5ml, emulsion is poured into frozen water rapidly, low-temperature setting 1h, 0.45um filter membrane
Filtering, obtains liquid phase P SO-DOX-NLC；
7) 15% mannitol+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
(2) testing result
1. specificity is tested
Chromatographic column：Syncronis C18250mm × 4.6mm, 5 μm)；Mobile phase：Methanol-acetic acid sodium buffer solution glacial acetic acid
Adjust pH3.6 (65:35)；Flow velocity:1.0mL/min；Ultraviolet detection wavelength:254nm；Column temperature：Room temperature；Sample size：20μL.
Method：Blank NLC (being not added with PSO and DOX) is simultaneously by above-mentioned steps, appropriate blank NLC and PSO-DOX- is measured
NLC adds mobile phase ultrasonic emulsion breaking, crosses 0.45 μm of filter membrane standby.And it is each appropriate in 10ml capacity to weigh a certain amount of PSO and DOX
In bottle, plus mobile phase ultrasonic dissolution, cross 0.45 μm of filter membrane and save backup.Precision weighs 5mgPSO and 5mgDOX and is placed in 100ml appearances
In measuring bottle, mobile phase ultrasonic dissolution, and constant volume are added, 50 μ g/ml PSO-DOX mixed reference substance solutions are configured to.By above-mentioned
Five solution difference sample introduction of condition configuration observes the disturbed condition of each composition.
As a result as in figs. 1 a-e.By the visible PSO of Figure 1A-Fig. 1 E, DOX, blank NLC, PSO-DOX-NLC and PSO-
It is noiseless between DOX each sample, separating degree R>1.5.The method for illustrating the present invention is feasible.
2. linear relationship is investigated
Method：Precision measures reference substance PSO (or reference substance DOX) solution in right amount in 10ml volumetric flasks, plus flowing mixes
Solution is diluted to scale, is configured to the series concentration that concentration is 1.0,2.0,4.0,8.0,16.0,32.0 μ g/ml, and HPLC surveys its peak
Area, using peak area as ordinate (Y), reference substance concentration is abscissa (X), carries out linear regression.
As a result：PSO calibration curve equation Y=1.1 × 105X+14742 (r=0.9998)；DOX calibration curve equation
Y=54195X+1877.9 (r=0.9999), shows that PSO and the DOX linear relationship in the range of 1~32 μ g/mL are good.
3. Precision Experiment
Method：Precision measures reference substance PSO (or reference substance DOX) solution in right amount in 10ml volumetric flasks, and configuration concentration is
4th, 16,32 μ g/ml basic, normal, high three strength solutions, HPLC is determined, respectively at parallel determination 5 times in 1 day, METHOD FOR CONTINUOUS DETERMINATION 5
My god, calculate in a few days with day to day precision RSD.
It the results are shown in Table 1.Show that precision is good by table 1.
In table 1 day and day to day precision result (n=5)
4. the rate of recovery is tested
Method：Take 1ml blank NLC, be separately added into reference substance PSO (or reference substance DOX) solution 0.8,3.2,6.4ml in
10ml volumetric flasks, the sample of basic, normal, high three concentration is made into mobile phase, and ultrasonic emulsion breaking, 0.45 μm of membrane filtration, 20 μ L enter
Sample, calculates the rate of recovery.
As a result：As a result such as table 2.As a result show PSO and DOX three strength solution rate of recovery 95%~103% it
Between, RSD<3%, show that this method rate of recovery is good.
The rate of recovery experimental result (n=3) of table 2
5. stability experiment
Method：Reference substance PSO (or reference substance DOX) solution is taken to configure 8 μ g/ml PSO-DOX solution in right amount, respectively at 0,
2nd, 4,6,12, sample introduction in 24h, calculates RSD.
As a result show, PSO and the DOX RSD in 24h are respectively 0.79% and 1.55%, show that solution is stable in 24h.
6. entrapment efficiency determination method
Method：Experiment surveys its envelop rate using ultrafiltration centrifugal process.Appropriate PSO-DOX-NLC solution is taken in ultra-filtration centrifuge tube
In, 12000r/min centrifugation 20min take 20ul filtrate injecting chromatographs, determine free drug amount (WF)；Separately take 1.0mlPSO-
DOX-NLC solution is in 10ml volumetric flasks, plus mobile phase constant volume, demulsification, 0.45um membrane filtrations, takes 20ul filtrates to inject chromatogram
Instrument, calculates total dose (WT).Envelop rate=(WT-WF)/WT× 100%.
As a result：PSO envelop rates are that 76.56%, DOX envelop rates are 89.48%.
7. morphology is investigated
Method：Take PSO-DOX-NLC appropriate on the copper mesh with carbon film, dyed, dried naturally rearmounted with 2% phosphotungstic acid
In observation under transmission electron microscope.
As a result it is well dispersed as shown in Fig. 2 from Figure 2 it can be seen that nanoparticle is circular or similar round entity bead, substantially without
It is adhered.
8. particle diameter and Zeta potential are determined
Method：Take PSO-DOX-NLC to be diluted in right amount, using Zetasizer nanometer lasers particle size analyzer carry out particle diameter with
The measure of current potential.
As a result as shown in Figure 3 and Figure 4, from Fig. 3 and Fig. 4, the particle diameter of nanoparticle is for (128.7 ± 1.8) nm, PDI
0.22 ± 0.01 (n=3), current potential is (- 20.17 ± 0.31) mV, it is seen that the nano-carrier particle diameter of preparation is small, is uniformly dispersed, tool
There is preferable stability.
9. the foundation of release profiles
Method：5ml PSO-DOX-NLC are taken in the bag filter that molecular cut off is 10KDa, while configuring suitable concentration
PSO and DOX solution 5ml in bag filter, bag filter is placed in the 150mlPBS dissolution mediums containing 5% Tween 80, rotating speed
50r/min, (37 ± 0.5) DEG C dynamic dialysis, respectively at 0.5,1,2,4,8,12,24,36h point in time sampling 1m, add simultaneously
Synthermal dissolution medium 1ml.Dissolution medium drug concentration is determined using HPLC methods, and calculates preparation.
As a result as fig. 5 a and fig. 5b, from Fig. 5 A and Fig. 5 B, models fitting is carried out to it, meets Bidirectional power
Equation.PSO and DOX drug solutions have discharged complete substantially in 12h, and PSO is in preceding 8h internal memories in PSO-DOX-NLC nano-solutions
In burst effect, DOX is then sustained preferably, therefore the nano-carrier has good sustained release performance.
Embodiment 2 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
Preparation method is as follows：
1) by 3mg psoralens (PSO), 56mg solid lipids (mountain Yu's acid glyceride), 14mg liquid fatty (meat
Isopropyl myristate) and 140mg fat-soluble emulsifiers (soybean lecithin) sequentially add in 20ml ethanol, 78
Dissolved by heating in DEG C water-bath, ultrasonic disperse is mixed, and is used as organic phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (PLURONICS F87) are dissolved in 1ml waters for injection, 78
Dissolved by heating in DEG C water-bath, stir, be used as interior aqueous phase；
3) 520mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under 1000r/min stirrings, 5ml colostrums are slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to 5ml, emulsion is poured into frozen water rapidly, low-temperature setting 1h, 0.45um filter membrane mistake
Filter, obtains liquid phase P SO-DOX-NLC；
7) 15% mannitol+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
PSO-DOX-NLC is circular, similar round entity ball, and PSO envelop rates are reachable up to 78.8%, DOX envelop rates
84.0%.
Embodiment 3 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
Preparation method is as follows：
1) by 3mg psoralens (PSO), 56mg solid lipids (weight ratio, glycerin monostearate：Mountain Yu's acid glyceride
=1：1), 14mg liquid fatties (oleic acid) and 70mg fat-soluble emulsifiers (soybean lecithin) are sequentially added in 20ml ethanol, and 78
Dissolved by heating in DEG C water-bath, ultrasonic disperse is mixed, and is used as organic phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (PLURONICS F87) are dissolved in 1ml waters for injection, 78
Dissolved by heating in DEG C water-bath, stir, be used as interior aqueous phase；
3) 520mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under 1000r/min stirrings, 7ml colostrums are slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to 5ml, emulsion is poured into frozen water rapidly, low-temperature setting 0.5h, 0.45um filter membrane
Filtering, obtains liquid phase P SO-DOX-NLC；
7) 15% sucrose+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
PSO-DOX-NLC is circular, similar round entity ball, and PSO envelop rates are reachable up to 74.8%, DOX envelop rates
79.3%.
Embodiment 4 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
Preparation method is as follows：
1) by 3mg psoralens (PSO), 80mg solid lipids (weight ratio, GMS：Precirol ATO5=1:1)、20mg
Liquid fatty (Miglyol 812N (LABRATAC LIPOPHIE WL1349)) and 150mg fat-soluble emulsifier (soybean
Lecithin) sequentially add in 20ml ethanol, dissolved by heating in 78 DEG C of water-baths, ultrasonic disperse is mixed, and is used as organic phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (PLURONICS F87) are dissolved in 1ml waters for injection, 78
Dissolved by heating in DEG C water-bath, stir, be used as interior aqueous phase；
3) 400mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under 1000r/min stirrings, 7ml colostrums are slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to about 10ml, emulsion is poured into frozen water rapidly, low-temperature setting 1.5h, 0.45um filter
Membrane filtration, obtains liquid phase P SO-DOX-NLC；
7) 15% sucrose+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
PSO-DOX-NLC is circular, similar round entity ball, and PSO envelop rates are reachable up to 72.2%, DOX envelop rates
86.7%.
Embodiment 5 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
Preparation method is as follows：
1) by 3mg psoralens (PSO), 104mg solid lipids (weight ratio, GMS：Precirol ATO5=1:1)、
26mg liquid fatties (Miglyol 812N (LABRATAC LIPOPHIE WL1349)) and 130mg fat-soluble emulsifiers
(soybean lecithin) is sequentially added in 20ml ethanol, is dissolved by heating in 78 DEG C of water-baths, and ultrasonic disperse is mixed, and is used as organic phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (NaTDC) are dissolved in 1ml waters for injection, 78 DEG C
Dissolved by heating in water-bath, stir, be used as interior aqueous phase；
3) 520mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under 1000r/min stirrings, 5ml colostrums are slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to about 7ml, emulsion is poured into frozen water rapidly, low-temperature setting 1h, 0.45um filter membrane
Filtering, obtains liquid phase P SO-DOX-NLC；
7) 15% sucrose+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
PSO-DOX-NLC is circular, similar round entity ball, and PSO envelop rates are reachable up to 77.5%, DOX envelop rates
89.0%.
Embodiment 6 contains psoralen-adriamycin composite nanostructure lipid vector preparation (PSO-DOX-NLC)
Preparation method：
1) by 3mg psoralens (PSO), 120mg solid lipids (weight ratio, GMS：Precirol ATO5=1:1)、
30mg liquid fatties (Miglyol 812N (LABRATAC LIPOPHIE WL1349)) and 225mg fat-soluble emulsifiers
(soybean lecithin) is sequentially added in 20ml ethanol, is dissolved by heating in 78 DEG C of water-baths, and ultrasonic disperse is mixed, and is used as organic phase；
2) 3mg adriamycins (DOX), 25mg water soluble emulsifiers (PLURONICS F87) are dissolved in 1ml waters for injection, 78
Dissolved by heating in DEG C water-bath, stir, be used as interior aqueous phase；
3) 400mg surfactants (Tween 80) are dissolved in 20ml waters for injection, dissolved by heating in 78 DEG C of water-baths, stirring
Uniformly, as outer aqueous phase；
4) under 1000r/min stirrings, aqueous phase in 1ml is slowly injected into 20ml organic phases, stirring 10min is formed just
Breast；
5) under solid lipid stirring, 10ml colostrums are slowly injected into outside 20ml in aqueous phase, stirring and emulsifying formation emulsion；
6) when emulsion is concentrated by evaporation to about 5ml, emulsion is poured into frozen water rapidly, low-temperature setting 2h, 0.45um filter membrane
Filtering, obtains liquid phase P SO-DOX-NLC；
7) 15% sucrose+aqueous trehalose 1ml is added into 2ml liquid phase Ps SO-DOX-NLC, is freeze-dried, obtains solid phase
PSO-DOX-NLC。
PSO-DOX-NLC is circular, similar round entity ball, and PSO envelop rates are reachable up to 74.9%, DOX envelop rates
87.4%.
Claims (4)
1. one kind contains psoralen-adriamycin composite nanostructure lipid vector preparation, it is characterised in that described to contain Psoralen
Fat element-adriamycin composite nanostructure lipid vector preparation is sustained release preparation, and preparation method is as follows：
1）Psoralen, solid lipid, liquid fatty and fat-soluble emulsifier are sequentially added in organic solvent, 70-80 DEG C of water
Dissolved by heating in bath, ultrasonic disperse is mixed, and is used as organic phase；
2）Adriamycin and water soluble emulsifier are dissolved in appropriate water for injection, dissolved by heating in 70-80 DEG C of water-bath, stirring is equal
It is even, it is used as interior aqueous phase；
3）Surfactant is dissolved in appropriate water for injection, is dissolved by heating in 70-80 DEG C of water-bath, is stirred, be used as outer water
Phase；
4）Under agitation, interior aqueous phase is slowly injected into organic phase, stirs 5-15min, form colostrum；
5）Under agitation, colostrum is slowly injected into outer aqueous phase, stirring and emulsifying formation emulsion；
6）Poured into after emulsion is concentrated by evaporation in frozen water rapidly, 0.5 ~ 1.5h of low-temperature setting, micro porous filtration obtains liquid phase and contains Psoralen
Fat element-adriamycin composite nanostructure lipid vector preparation；
Wherein, following raw material is as follows by weight ratio：1-5 part of psoralen, 1-5 part of adriamycin, solid lipid 40-120
Part, 10-30 parts of liquid fatty, 35-350 parts of fat-soluble emulsifier, 25-250 parts of water soluble emulsifier, surfactant 100-
600 parts；
The solid lipid is one kind or two in glycerin monostearate, palmitic acid stearic acid ester of glycerol, Compritol 888 ATO
Plant the combination of the above；
The liquid fatty is one or two or more kinds of mixed in Miglyol 812N, isopropyl myristate or oleic acid
Close；
The fat-soluble emulsifier is soybean lecithin；
The water soluble emulsifier is poloxamer or one kind in NaTDC or two kinds of mixing；
Described surfactant is Tween 80.
2. one kind contains psoralen-adriamycin composite nanostructure lipid vector preparation, it is characterised in that described to contain Psoralen
Fat element-adriamycin composite nanostructure lipid vector preparation is sustained release preparation, and preparation method is as follows：
1）Psoralen, solid lipid, liquid fatty and fat-soluble emulsifier are sequentially added in organic solvent, 70-80 DEG C of water
Dissolved by heating in bath, ultrasonic disperse is mixed, and is used as organic phase；
2）Adriamycin and water soluble emulsifier are dissolved in appropriate water for injection, dissolved by heating in 70-80 DEG C of water-bath, stirring is equal
It is even, it is used as interior aqueous phase；
3）Surfactant is dissolved in appropriate water for injection, is dissolved by heating in 70-80 DEG C of water-bath, is stirred, be used as outer water
Phase；
4）Under agitation, interior aqueous phase is slowly injected into organic phase, stirs 5-15min, form colostrum；
5）Under agitation, colostrum is slowly injected into outer aqueous phase, stirring and emulsifying formation emulsion；
6）Poured into after emulsion is concentrated by evaporation in frozen water rapidly, 0.5 ~ 1.5h of low-temperature setting, micro porous filtration obtains liquid phase and contains Psoralen
Fat element-adriamycin composite nanostructure lipid vector preparation；
7）Addition frozen-dried protective agent solution in psoralen-adriamycin composite nanostructure lipid vector preparation is contained to liquid phase,
Dry, obtain solid phase and contain psoralen-adriamycin composite nanostructure lipid vector preparation；
Wherein, following raw material is as follows by weight ratio：1-5 part of psoralen, 1-5 part of adriamycin, solid lipid 40-120
Part, 10-30 parts of liquid fatty, 35-350 parts of fat-soluble emulsifier, 25-250 parts of water soluble emulsifier, surfactant 100-
600 parts；
The solid lipid is one kind or two in glycerin monostearate, palmitic acid stearic acid ester of glycerol, Compritol 888 ATO
Plant the combination of the above；
The liquid fatty is one or two or more kinds of mixed in Miglyol 812N, isopropyl myristate or oleic acid
Close；
The fat-soluble emulsifier is soybean lecithin；
The water soluble emulsifier is poloxamer or one kind in NaTDC or two kinds of mixing；
Described surfactant is Tween 80.
3. contain psoralen-adriamycin composite nanostructure lipid vector preparation, its feature as claimed in claim 1 or 2
It is：Step 6）Described in micro porous filtration be to use aperture for 0.22 μm or 0.45 μm of water system membrane filtration.
4. psoralen-adriamycin composite nanostructure the lipid vector preparation that contains described in claim 1 or 2 is preparing treatment
Application in antineoplastic.
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| CN108853056B (en) * | 2018-07-02 | 2021-09-14 | 天津中医药大学 | Folic acid targeted modification carried doxorubicin hydrochloride and gambogic acid nano-structure lipid carrier preparation and preparation method thereof |
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| CN111494619B (en) * | 2020-04-26 | 2022-03-18 | 南京农业大学 | Preparation method of squalene-based cationic nanostructured lipid carrier immunologic adjuvant |
| CN115813880B (en) * | 2022-04-02 | 2024-06-04 | 海南医学院 | Benzyl isothiocyanate-nanostructure lipid carrier and preparation method thereof |
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