CN106620697A - Nano flower-like palladium-gold coated betulinic acid liposome and preparation method thereof - Google Patents
Nano flower-like palladium-gold coated betulinic acid liposome and preparation method thereof Download PDFInfo
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- CN106620697A CN106620697A CN201611012105.4A CN201611012105A CN106620697A CN 106620697 A CN106620697 A CN 106620697A CN 201611012105 A CN201611012105 A CN 201611012105A CN 106620697 A CN106620697 A CN 106620697A
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Abstract
A nano flower-like palladium-gold coated betulinic acid liposome is a nano liposome formed by forming a nano flower-like palladium-gold alloy shell on the surface of a betulinic acid nano liposome, with the particle size of about 80-100 nm and the near infrared absorption wavelength of 500-900 nm. A preparation method mainly comprises steps as follows: palladium nanoparticles are obtained by reducing a palladium chloride solution with ascorbic acid, the palladium nanoparticles and the betulinic acid nano liposome modified with glutathione are subjected to shaded incubation under the mild condition, then a gold chloride solution and a hydroxylamine hydrochloride solution are added, and shaded incubation is performed for 4-9 h. The preparation method is simple to operate, easy to control and high in repeatability; the prepared nano flower-like palladium-gold coated betulinic acid liposome has good photo-thermal conversion performance, makes medicine release controllable, is more effective and low-toxic and can combine photo-thermal therapy and chemotherapy to treat cancer collaboratively.
Description
Technical field
The invention belongs to pharmaceutical technology field, more particularly to a kind of nano liposomes and preparation method thereof.
Background technology
Cancer is one of great malignant disease of current threat human health, and the classic chemotherapy medicine for being used at present is deposited
In serious toxic and side effect, and the treatment for appearing as cancer of nanometer technology provides new approaches.Wherein, liposome is a kind of nothing
Poison, biocompatibility is fine, the nano medicament carrying system of great prospect.But, in order to reach more preferable therapeutic effect, conventional fat
Such as also there are some defects in plastid, unstable, uncontrollability of targeting difference and insoluble drug release etc..
In recent years, various optothermal materials are gradually applied in biomedicine field.Wherein, the nanometer of near infrared light response
Investigation of materials is the most extensive, for example noble metal nanometer material (gold nano-material), Graphene, changes on CNT and rare earth
Luminescent material.Between 650-900nm, tissue absorbs and scatters little, Jin Erneng to " biological tissue's window " of near infrared light to it
Enough reach tumor tissues depths and normal tissue has no toxic side effect.The operation principle of these optothermal materials is that luminous energy is good
Heat energy is converted into, so as to suppress the growth of tumour cell of temperature-sensitive.But, simple photo-thermal therapy is not enough to kill tumour cell.
Therefore, developing new therapeutic modality and research becomes at this stage that field of cancer treatment is urgently for the efficient joint treatment of cancer
Problem to be solved.
The content of the invention
It is an object of the invention to provide a kind of simple to operate, easy control of reaction system, repeatability are high, the nano liposomes of preparation
Be easier to tumor locus accumulation, anisotropy it is relatively good, more efficiently, the white birch fat that coats of nano flower-like palladium-gold of low toxicity
Sour liposome and preparation method thereof.
The present invention mainly uses the betulic acid nano liposomes that alcohol injection prepares glutathione modification；Then will
The betulic acid nano liposomes of glutathione modification mix with Pd nano particle solution, and after lucifuge incubation chlorogold solution is added
And hydroxylamine hydrochloride solution, obtain that insoluble drug release is controllable and white birch fat of nano flower-like palladium of thermotherapy/chemotherapy Synergistic treatment-gold cladding
Sour liposome.
The betulic acid liposome of the present invention is a kind of in betulic acid nano liposomes surface formation nano flower-like palladium-gold
The nano liposomes of alloy shell, its particle diameter is 80-100nm, and near infrared absorption wavelength is 500-900nm.
The preparation method of above-mentioned betulic acid liposome is as follows：
1st, the betulic acid nano liposomes of glutathione modification are prepared
(1) mass ratio of each raw material is：Soybean lecithin：Cholesterol：Betulic acid=12～20:1.3～2.1:1,
It is 14.3～23.1mg to add the summation of above-mentioned three kinds of Solute mass by every milliliter of absolute ethyl alcohol, adds absolute ethyl alcohol and ultrasound,
Ultrasonic power is 40～80W, and ultrasonic time is 10-20min so as to is completely dissolved and obtains lipid soln；
(2) take glutathione to be dissolved in deionized water, obtain glutathione solution, in every ml deionized water 0.3 is contained
～1mg glutathione；Volume ratio with lipid soln and glutathione solution is as 1:1.5～3.5, magnetic agitation and temperature control
System in the glutathione solution for being added dropwise to the lipid soln that step (1) is obtained, obtains liposome and hangs at 38～47 DEG C
Liquid；
(3) at 28～35 DEG C, 100～130rpm carries out magnetic force in 3～7 hours and stirs the Liposomal suspensions for obtaining step (2)
Mix, remove the absolute ethyl alcohol in Liposomal suspensions, obtain the betulic acid nano liposomes of glutathione modification.
2nd, the betulic acid nano liposomes of nano flower-like palladium-billon cladding are prepared
(1) volume ratio with palladium chloride solution and ascorbic acid solution is as 100:1～4 ratio, is 50 by molar concentration
The ascorbic acid solution of～100mM, in being added drop-wise to the palladium chloride solution that molar concentration is 0.5～1.5mM, is well mixed rapidly,
Until color is changed into light green color, that is, Pd nano particle solution is obtained, be placed in storage at normal temperature, it is standby；
(2) the Pd nano particle solution for obtaining step (1) is received with the betulic acid of glutathione modification obtained in step 1
Mizhi plastid presses 1:1～5 volume ratio is well mixed, and is placed in shaking table, and 25～30 DEG C, 85～115rpm, lucifuge incubation 8～16 is little
When, that is, obtain the betulic acid nano liposomes of Pd nano particle cladding；
(3) it is 1 by the volume ratio of chlorogold solution and the betulic acid nano liposomes of Pd nano particle cladding:0.5～
1.5 ratio, adds molar concentration to be 2 in the betulic acid nano liposomes of the Pd nano particle cladding obtained to step (2)
～8mM chlorogold solutions, fully mix；Then to adding molar concentration to be 50～100mM hydroxylamine hydrochloride solutions in mixed liquor, fill
Divide and be well mixed, hydroxylamine hydrochloride solution is 1～5 with the volume ratio of chlorogold solution：10, it is placed on shaking table, 25～30 DEG C, 85
～115rpm, is incubated 6～12 hours, and the betulic acid liposome of nano flower-like palladium-gold cladding is obtained.
On the basis of conventional medicine liposome, the betulic acid liposome of glutathione modification is prepared for.Nano flower-like
There is near infrared absorption in place in 500-900nm width wave-length coverages in the betulic acid nano liposomes of palladium-billon cladding, so as to
With good photothermal conversion performance.Lipid temperature is raised by near infrared light, and then promotes package-contained medicine in target
The controlled release in site, meanwhile, it is capable to cause tumour cell that heat fusion and death occur using the thermally-induced intensification of light, it is seen then that
The drug-loading system by photo-thermal therapy in combination with chemotherapy, can realize the synergistic therapeutic effect of cancer, with single smooth technique for hyperthermia or
Chemotherapy is compared more efficiently, and to the Small side effects of human body.
The present invention has the advantage that compared with prior art：
1st, complete to prepare under normal temperature, normal pressure and temperate condition, simple to operate, easy control of reaction system, repeatability is high.
2nd, the particle diameter of the betulic acid liposome of the nano flower-like palladium for preparing-gold cladding is easier to be accumulated in tumor locus,
Reach more preferable therapeutic effect.
3rd, the betulic acid liposome that the nano flower-like palladium-gold for preparing is coated is compared with existing spherical shell shape pharmaceutical carrier,
Anisotropy is relatively good, and it has near infrared absorption in place in 500-900nm width wave-length coverages, with good photothermal conversion
Can, make the antineoplastic betulic acid for containing realize controlled release.
4th, the betulic acid liposome of the nano flower-like palladium for preparing-gold cladding, can be by light under near-infrared laser irradiation
Heat cure is combined Synergistic treatment cancer with chemotherapy, compared with single smooth technique for hyperthermia or chemotherapy more efficiently, low toxicity.
Description of the drawings
Fig. 1 is the betulic acid liposome transmission electron microscopy of nano flower-like palladium obtained in the embodiment of the present invention 1-gold cladding
Mirror figure.
Fig. 2 is the betulic acid liposomal particle size distribution map of nano flower-like palladium obtained in the embodiment of the present invention 2-gold cladding.
Fig. 3 is the betulic acid liposome uv-vis spectra of nano flower-like palladium obtained in the embodiment of the present invention 3-gold cladding
Figure.
Fig. 4 is the photothermal conversion of the betulic acid liposome of nano flower-like palladium obtained in the embodiment of the present invention 3-gold cladding
Figure.
Specific embodiment
Embodiment 1
4mg betulic acids (being purchased from Nanjing Ze Lang bio tech ltd), 5.2mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 48mg soybean lecithins (be purchased from Shanghai Advanced viecle Technology Co., Ltd.), be dissolved in 4mL without
In water-ethanol and ultrasound is carried out, ultrasonic power 40W, ultrasonic 10min obtain lipid soln；Take 6mg glutathione and be dissolved in 20mL
In deionized water, glutathione solution is obtained；38 DEG C of magnetic agitations, the 15mL gluathiones that 10mL lipid solns are added dropwise to
In peptide solution, Liposomal suspensions are obtained；Subsequently by the Liposomal suspensions for obtaining at 28 DEG C, 100rpm, magnetic agitation 3 hours, with
The absolute ethyl alcohol in Liposomal suspensions is removed, the betulic acid nano liposomes of glutathione modification are obtained；
10 μ L molar concentrations are added drop-wise to into the palladium bichloride that 1000 μ L molar concentrations are 0.5mM for the ascorbic acid solution of 50mM
In solution, it is well mixed rapidly, until color is changed into light green color, that is, obtains Pd nano particle solution, is placed in storage at normal temperature, it is standby
With；By the Pd nano particle solution for obtaining and the betulic acid nano liposomes of glutathione modification by 1:1 volume ratio mixing
Uniformly, shaking table is placed in, 25 DEG C, 85rpm, lucifuge is incubated 8 hours, that is, obtain the betulic acid nano-lipid of Pd nano particle cladding
Body.
The betulic acid nano liposomes of 500 μ L Pd nano particles cladding are taken, 1000 μ L molar concentrations is added for the chlorine of 2mM
Change gold solution, fully mix, the hydroxylamine hydrochloride solution that subsequently 100 μ L molar concentrations will be added to be 50mM in mixed liquor, fully
It is well mixed, is placed on shaking table, 25 DEG C, 85rpm is incubated 6 hours, that is, obtain the betulic acid of nano flower-like palladium-gold cladding
Liposome.
The betulic acid liposome of nano flower-like palladium-gold cladding is characterized using transmission electron microscope, such as Fig. 1 institutes
Show, it can be seen that form flower-shaped palladium-billon housing, pattern rule on betulic acid nano liposomes surface.
Embodiment 2
4mg betulic acids (being purchased from Nanjing Ze Lang bio tech ltd), 6.8mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 64mg soybean lecithins (be purchased from Shanghai Advanced viecle Technology Co., Ltd.), be dissolved in 4mL without
In water-ethanol and ultrasonic 60W, 15min are carried out, obtain lipid soln.Take 12mg glutathione to be dissolved in 20mL deionized waters,
Obtain glutathione solution；42 DEG C of magnetic agitations, in the 25mL glutathione solutions that 10mL lipid solns are added dropwise to, obtain
To Liposomal suspensions；Subsequently by the Liposomal suspensions for obtaining at 31 DEG C, 115rpm, magnetic agitation 5 hours is hanged with removing liposome
Absolute ethyl alcohol in liquid, obtains the betulic acid nano liposomes of glutathione modification；
25 μ L molar concentrations are added drop-wise to into the palladium bichloride that 1000 μ L molar concentrations are 1mM for the ascorbic acid solution of 75mM molten
In liquid, it is well mixed rapidly, until color is changed into light green color, that is, obtains Pd nano particle solution, is placed in storage at normal temperature, it is standby；
By the Pd nano particle solution for obtaining and the betulic acid nano liposomes of glutathione modification by 1:3 volume ratio mixing is equal
It is even, shaking table is placed in, 27 DEG C, 105rpm, lucifuge is incubated 12 hours, that is, obtain the betulic acid nano-lipid of Pd nano particle cladding
Body；
The betulic acid nano liposomes of 1000 μ L Pd nano particles cladding are taken, 1000 μ L molar concentrations is added for 5mM's
Chlorogold solution, fully mixes.The hydroxylamine hydrochloride solution that subsequently 300 μ L molar concentrations will be added to be 75mM in mixed liquor, fills
Divide and be well mixed, be placed on shaking table, 27 DEG C, 105rpm is incubated 9 hours, that is, obtain the white birch fat of nano flower-like palladium-gold cladding
Sour liposome.
The betulic acid liposome of nano flower-like palladium-gold cladding is characterized using laser particle analyzer, as shown in Fig. 2
Understand that its particle diameter, about in 98nm, is more beneficial for the accumulation in tumor locus from granularmetric analysis result, reach preferable therapeutic effect.
Embodiment 3
4mg betulic acids (being purchased from Nanjing Ze Lang bio tech ltd), 8.4mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 80mg soybean lecithins (be purchased from Shanghai Advanced viecle Technology Co., Ltd.), be dissolved in 4mL without
In water-ethanol and ultrasonic 80W, 20min are carried out, obtain lipid soln；Take 20mg glutathione to be dissolved in 20mL deionized waters,
Obtain glutathione solution；47 DEG C of magnetic agitations, in the 35mL glutathione solutions that 10mL lipid solns are added dropwise to, obtain
To Liposomal suspensions.Subsequently by the Liposomal suspensions for obtaining at 35 DEG C, 130rpm, magnetic agitation 7 hours is hanged with removing liposome
Absolute ethyl alcohol in liquid, obtains the betulic acid nano liposomes of glutathione modification；
It is the ascorbic acid solution of 100mM by 40 μ L molar concentrations, is added drop-wise to the chlorination that 1000 μ L molar concentrations are 1.5mM
In palladium solution, it is well mixed rapidly, until color is changed into light green color, that is, obtains Pd nano particle solution, is placed in storage at normal temperature, it is standby
With；By the Pd nano particle solution for obtaining and the betulic acid nano liposomes of glutathione modification by 1:5 volume ratio mixing
Uniformly, shaking table is placed in, 30 DEG C, 115rpm, lucifuge is incubated 16 hours, that is, obtain the betulic acid nanometer fat of Pd nano particle cladding
Plastid；
The betulic acid nano liposomes of 1500 μ L Pd nano particles cladding are taken, 1000 μ L molar concentrations is added for 8mM's
Chlorogold solution, fully mixes；The hydroxylamine hydrochloride solution that subsequently 500 μ L molar concentrations will be added to be 100mM in mixed liquor, fills
Divide and be well mixed, be placed on shaking table, 30 DEG C, 115rpm is incubated 12 hours, that is, obtain the white birch of nano flower-like palladium-gold cladding
Resin acid liposome.
The betulic acid liposome of nano flower-like palladium-gold cladding is characterized using ultraviolet-visual spectrometer, such as Fig. 3 institutes
Show, it can be seen that after nano flower-like palladium-billon housing is formed, it is near that it locates presence in 500-900nm width wave-length coverages
INFRARED ABSORPTION；Meanwhile, the betulic acid liposome photothermal conversion of nano flower-like palladium-gold cladding is detected, as shown in figure 4,
At 10 minutes, this nano medicament carrying system temperature can be increased to 51.3 DEG C, thus with good photothermal conversion performance.
Claims (2)
1. the betulic acid liposome that a kind of nano flower-like palladium-gold is coated, it is characterised in that：It is one kind in betulic acid nanometer
Surface of liposome forms the nano liposomes of nano flower-like palladium-billon housing, and its particle size is about 80-100nm, near-infrared
Absorbing wavelength is 500-900nm.
2. the preparation method of the betulic acid liposome of the nano flower-like palladium of claim 1-gold cladding, it is characterised in that：It is wrapped
Include following steps：
(1) the betulic acid nano liposomes of glutathione modification are prepared
1. the mass ratio of each raw material is：Soybean lecithin：Cholesterol：Betulic acid=12～20:1.3～2.1:1, by every milli
It is 14.3～23.1mg to rise absolute ethyl alcohol and add the summation of above-mentioned three kinds of Solute mass, adds absolute ethyl alcohol and ultrasound, ultrasonic work(
Rate is 40～80W, and ultrasonic time is 10-20min so as to is completely dissolved and obtains lipid soln；
2. take glutathione to be dissolved in deionized water, obtain glutathione solution, in every ml deionized water 0.3～1mg is contained
Glutathione；Volume ratio with lipid soln and glutathione solution is as 1:1.5～3.5, magnetic agitation and temperature control exists
38～47 DEG C, in the glutathione solution that the lipid soln that 1. step obtains is added dropwise to, obtain Liposomal suspensions；
3. at 28～35 DEG C, 100～130rpm carries out magnetic agitation in 3～7 hours to the Liposomal suspensions for 2. step being obtained, and removes
The absolute ethyl alcohol gone in Liposomal suspensions, obtains the betulic acid nano liposomes of glutathione modification；
(2) the betulic acid nano liposomes of nano flower-like palladium-billon cladding are prepared
1. the volume ratio with palladium chloride solution and ascorbic acid solution is as 100:1～4 ratio, by molar concentration be 50～
The ascorbic acid solution of 100mM, in being added drop-wise to the palladium chloride solution that molar concentration is 0.5～1.5mM, is well mixed rapidly, directly
It is changed into light green color to color, that is, obtains Pd nano particle solution, is placed in storage at normal temperature, it is standby；
2. by step 1. in the betulic acid nanometer of glutathione modification that obtains with step (1) of the Pd nano particle solution that obtains
Liposome presses 1:1～5 volume ratio is well mixed, and is placed in shaking table, and 25～30 DEG C, 85～115rpm, lucifuge incubation 8～16 is little
When, that is, obtain the betulic acid nano liposomes of Pd nano particle cladding；
3. with chlorogold solution and the volume ratio 10 of the betulic acid nano liposomes of Pd nano particle cladding:5～15 ratio,
Molar concentration is added to be 2～8mM chloraurides in the betulic acid nano liposomes of the Pd nano particle cladding 2. obtained to step
Solution, fully mixes.Molar concentration subsequently will be added in mixed liquor to be 50～100mM hydroxylamine hydrochloride solutions, be sufficiently mixed
It is even, and chlorogold solution and the volume ratio of hydroxylamine hydrochloride solution are 10:1～5, it is placed on shaking table, 25～30 DEG C, 85～
115rpm, is incubated 6～12 hours, that is, obtain the betulic acid liposome of nano flower-like palladium-gold cladding.
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| CN116376351A (en) * | 2023-04-07 | 2023-07-04 | 广东炎墨方案科技有限公司 | Preparation method of composite resin-based solder resist ink |
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