CN106620697B - A kind of betulic acid liposome and preparation method thereof of nano flower-like palladium-gold cladding - Google Patents
A kind of betulic acid liposome and preparation method thereof of nano flower-like palladium-gold cladding Download PDFInfo
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Abstract
A kind of betulic acid liposome of nano flower-like palladium-gold cladding, it is a kind of in betulic acid nano liposomes surface formation nano flower-like palladium-billon shell nano liposomes, and particle size is about 80-100nm, and near infrared absorption wavelength is 500-900nm；Preparation method is mainly: restoring palladium chloride solution using ascorbic acid, obtain Pd nano particle, and then carry out being protected from light incubation in a mild condition with the betulic acid nano liposomes of glutathione modification, chlorogold solution and hydroxylamine hydrochloride solution is then added, is protected from light incubation 4~9 hours.Preparation method of the present invention is easy to operate, reacts easy to control, repeated height；The betulic acid liposome of the nano flower-like palladium of preparation-gold cladding has good photothermal conversion performance, keeps drug release controllable, more high-efficiency low-toxicity, can by photo-thermal therapy in conjunction with chemotherapy synergistic treatment cancer.
Description
Technical field
The invention belongs to pharmaceutical technology field, in particular to a kind of nano liposomes and preparation method thereof.
Background technique
Cancer is one of great malignant disease of current threat human health, and used classic chemotherapy drug is deposited at present
In serious toxic side effect, and the appearance of nanotechnology provides new approaches for the treatment of cancer.Wherein, liposome is a kind of nothing
Poison, biocompatibility is fine, the nano medicament carrying system of great prospect.But in order to reach better therapeutic effect, conventional rouge
There is also some defects, such as unstable, targeting difference and the uncontrollability of drug release etc. for plastid.
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, such as noble metal nanometer material (gold nano-material), graphene, converts on carbon nanotube and rare earth
Luminescent material.For " the biological tissue's window " of near infrared light between 650-900nm, tissue absorbs to it and scatters small, Jin Erneng
Enough reach tumor tissues depths and normal tissue has no toxic side effect.The working principle of these optothermal materials is that luminous energy is good
It is converted into thermal energy, to inhibit the growth of tumour cell of temperature-sensitive.But simple photo-thermal therapy is not enough to kill tumour cell.
Therefore, novel therapeutic modality and research is developed to treat as field of cancer treatment at this stage urgently for the efficient joint of cancer
Problem to be solved.
Summary of the invention
The purpose of the present invention is to provide a kind of easy to operate, easy to control, repeated height of reaction, the nano liposomes of preparation
It is easier in the white birch rouge that tumor locus accumulates, anisotropy is relatively good, nano flower-like palladium more efficiently, less toxic-gold coats
Sour liposome and preparation method thereof.
The present invention is mainly the betulic acid nano liposomes that glutathione modification is prepared using alcohol injection；Then will
The betulic acid nano liposomes of glutathione modification are mixed with Pd nano particle solution, and chlorogold solution is added after being protected from light incubation
And hydroxylamine hydrochloride solution, obtain drug release controllably and thermotherapy/chemotherapy synergistic treatment nano flower-like palladium-gold cladding white birch rouge
Sour liposome.
Betulic acid liposome of the invention is a kind of in betulic acid nano liposomes surface formation nano flower-like palladium-gold
The nano liposomes of alloy shell, partial size 80-100nm, near infrared absorption wavelength are 500-900nm.
Above-mentioned betulic acid liposome the preparation method is as follows:
1, the betulic acid nano liposomes of glutathione modification are prepared
(1) mass ratio of each raw material are as follows: soybean lecithin: cholesterol: betulic acid=12~20:1.3~2.1:1,
It is 14.3~23.1mg by the summation that above-mentioned three kinds of Solute mass are added in every milliliter of dehydrated alcohol, dehydrated alcohol and ultrasound is added,
Ultrasonic power is 40~80W, and ultrasonic time 10-20min makes it completely dissolved to obtain lipid soln；
(2) it takes glutathione to be dissolved in deionized water, obtains glutathione solution, contain 0.3 in every ml deionized water
~1mg glutathione；Volume ratio with lipid soln and glutathione solution is 1:1.5~3.5, magnetic agitation and temperature control
It makes in the glutathione solution that at 38~47 DEG C, the lipid soln that step (1) obtains is added dropwise to, it is outstanding to obtain liposome
Liquid；
(3) Liposomal suspensions for obtaining step (2) are stirred in 28~35 DEG C, 100~130rpm, 3~7 hours progress magnetic force
It mixes, removes the dehydrated alcohol in Liposomal suspensions, obtain the betulic acid nano liposomes of glutathione modification.
2, nano flower-like palladium-billon cladding betulic acid nano liposomes are prepared
(1) it is the ratio of 100:1~4 with the volume ratio of palladium chloride solution and ascorbic acid solution, is 50 by molar concentration
The ascorbic acid solution of~100mM is added drop-wise in the palladium chloride solution that molar concentration is 0.5~1.5mM, is uniformly mixed rapidly,
Until color becomes light green color to get to Pd nano particle solution, it is placed in storage at normal temperature, it is spare；
(2) betulic acid that the Pd nano particle solution that step (1) obtains is modified with glutathione made from step 1 is received
Mizhi plastid is uniformly mixed by the volume ratio of 1:1~5, is placed in shaking table, 25~30 DEG C, it is small to be protected from light incubation 8~16 by 85~115rpm
When to get to Pd nano particle cladding betulic acid nano liposomes；
(3) by chlorogold solution and Pd nano particle cladding betulic acid nano liposomes volume ratio be 1:0.5~
1.5 ratio, it is 2 that molar concentration, which is added, into the betulic acid nano liposomes for the Pd nano particle cladding that step (2) obtain
~8mM chlorogold solution, mixes well；It is 50~100mM hydroxylamine hydrochloride solution then to molar concentration is added in mixed liquor, fills
It point being uniformly mixed, the volume ratio of hydroxylamine hydrochloride solution and chlorogold solution is 1~5:10, place it in shaking table, 25~30 DEG C, 85
~115rpm is incubated for 6~12 hours, and nano flower-like palladium-gold cladding betulic acid liposome is made.
On the basis of conventional medicine liposome, it is prepared for the betulic acid liposome of glutathione modification.Nano flower-like
There are near infrared absorptions in 500-900nm wide wave-length coverage for palladium-billon cladding betulic acid nano liposomes, thus
With good photothermal conversion performance.Lipid temperature is increased by near infrared light, and then promotes package-contained drug in target
The controlled release in site, meanwhile, it is capable to cause tumour cell that heat fusion and death occurs using heating caused by photo-thermal, it is seen then that
The drug-loading system can combine photo-thermal therapy with chemotherapy, realize the synergistic therapeutic effect of cancer, with single smooth technique for hyperthermia or
Chemotherapy is efficient compared to more, and to the Small side effects of human body.
The invention has the following advantages over the prior art:
1, preparation is completed under room temperature, normal pressure and temperate condition, it is easy to operate, react easy to control, repeatability is high.
2, the partial size of the nano flower-like palladium prepared-gold cladding betulic acid liposome is easier to accumulate in tumor locus,
Reach better therapeutic effect.
3, prepare nano flower-like palladium-gold coat betulic acid liposome compared with existing spherical shell shape pharmaceutical carrier,
Anisotropy is relatively good, and there are near infrared absorptions in 500-900nm wide wave-length coverage, has good photothermal conversion
Can, so that the anti-tumor drug betulic acid contained is realized controlled release.
4, the nano flower-like palladium prepared-gold cladding betulic acid liposome, can be by light under near-infrared laser irradiation
Heat cure synergistic treatment cancer in conjunction with chemotherapy, more efficient compared with single smooth technique for hyperthermia or chemotherapy, low toxicity.
Detailed description of the invention
Fig. 1 is the palladium of nano flower-like made from the embodiment of the present invention 1-gold cladding betulic acid liposome transmission electron microscopy
Mirror figure.
Fig. 2 is the palladium of nano flower-like made from the embodiment of the present invention 2-gold cladding betulic acid liposomal particle size distribution map.
Fig. 3 is the palladium of nano flower-like made from the embodiment of the present invention 3-gold cladding betulic acid liposome uv-vis spectra
Figure.
Fig. 4 is the photothermal conversion of the palladium of nano flower-like made from the embodiment of the present invention 3-gold cladding betulic acid liposome
Figure.
Specific embodiment
Embodiment 1
4mg betulic acid (being purchased from Nanjing Ze Lang Biotechnology Co., Ltd), 5.2mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 48mg soybean lecithin (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；6mg glutathione is taken to be dissolved in 20mL
In deionized water, glutathione solution is obtained；38 DEG C of magnetic agitations, the 15mL gluathione that 10mL lipid soln is added dropwise to
In peptide solution, Liposomal suspensions are obtained；Then by obtained Liposomal suspensions at 28 DEG C, 100rpm, magnetic agitation 3 hours, with
The dehydrated alcohol in Liposomal suspensions is removed, the betulic acid nano liposomes of glutathione modification are obtained；
The ascorbic acid solution that 10 μ L molar concentrations are 50mM is added drop-wise to the palladium chloride that 1000 μ L molar concentrations are 0.5mM
It in solution, is uniformly mixed rapidly, until color becomes light green color to get to Pd nano particle solution, is placed in storage at normal temperature, it is standby
With；Obtained Pd nano particle solution is mixed with the betulic acid nano liposomes that glutathione is modified by the volume ratio of 1:1
Uniformly, it is placed in shaking table, 25 DEG C, 85rpm, is protected from light and is incubated for 8 hours to get the betulic acid nano-lipid coated to Pd nano particle
Body.
The chlorine that 1000 μ L molar concentrations are 2mM is added in the betulic acid nano liposomes for taking 500 μ L Pd nano particles to coat
Change gold solution, mix well, then the hydroxylamine hydrochloride solution that 100 μ L molar concentrations are 50mM will be added into mixed liquor, sufficiently
It is uniformly mixed, places it in shaking table, 25 DEG C, 85rpm, be incubated for 6 hours to get the betulic acid coated to nano flower-like palladium-gold
Liposome.
Nano flower-like palladium-gold cladding betulic acid liposome is characterized using transmission electron microscope, such as Fig. 1 institute
Show, it can be seen that form flower-shaped palladium-billon shell, morphological rules on betulic acid nano liposomes surface.
Embodiment 2
4mg betulic acid (being purchased from Nanjing Ze Lang Biotechnology Co., Ltd), 6.8mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 64mg soybean lecithin (be purchased from Shanghai Advanced viecle Technology Co., Ltd.), be dissolved in 4mL without
In water-ethanol and ultrasonic 60W, 15min are carried out, obtains lipid soln.12mg glutathione is taken to be dissolved in 20mL deionized water,
Obtain glutathione solution；42 DEG C of magnetic agitations in the 25mL glutathione solution for being added dropwise to 10mL lipid soln, obtain
To Liposomal suspensions；It is outstanding to remove liposome then by obtained Liposomal suspensions at 31 DEG C, 115rpm, magnetic agitation 5 hours
Dehydrated alcohol in liquid obtains the betulic acid nano liposomes of glutathione modification；
It is molten that the ascorbic acid solution that 25 μ L molar concentrations are 75mM is added drop-wise to the palladium chloride that 1000 μ L molar concentrations are 1mM
It in liquid, is uniformly mixed rapidly, until color becomes light green color to get to Pd nano particle solution, is placed in storage at normal temperature, it is spare；
Obtained Pd nano particle solution is mixed with the betulic acid nano liposomes that glutathione is modified by the volume ratio of 1:3
It is even, it is placed in shaking table, 27 DEG C, 105rpm, is protected from light and is incubated for 12 hours to get the betulic acid nano-lipid coated to Pd nano particle
Body；
The betulic acid nano liposomes for taking 1000 μ L Pd nano particles to coat, it is 5mM's that 1000 μ L molar concentrations, which are added,
Chlorogold solution mixes well.Then the hydroxylamine hydrochloride solution that 300 μ L molar concentrations are 75mM will be added into mixed liquor, fills
Divide and be uniformly mixed, places it in shaking table, 27 DEG C, 105rpm, be incubated for 9 hours to get the white birch rouge coated to nano flower-like palladium-gold
Sour liposome.
Nano flower-like palladium-gold cladding betulic acid liposome is characterized using laser particle analyzer, as shown in Fig. 2,
It is more advantageous to the accumulation in tumor locus, reaches ideal therapeutic effect about in 98nm from its partial size known to granularmetric analysis result.
Embodiment 3
4mg betulic acid (being purchased from Nanjing Ze Lang Biotechnology Co., Ltd), 8.4mg cholesterol is (big purchased from Tianjin
Luxuriant chemical apparatuses supply station) and 80mg soybean lecithin (be purchased from Shanghai Advanced viecle Technology Co., Ltd.), be dissolved in 4mL without
In water-ethanol and ultrasonic 80W, 20min are carried out, obtains lipid soln；20mg glutathione is taken to be dissolved in 20mL deionized water,
Obtain glutathione solution；47 DEG C of magnetic agitations in the 35mL glutathione solution for being added dropwise to 10mL lipid soln, obtain
To Liposomal suspensions.It is outstanding to remove liposome then by obtained Liposomal suspensions at 35 DEG C, 130rpm, magnetic agitation 7 hours
Dehydrated alcohol in liquid obtains the betulic acid nano liposomes of glutathione modification；
The ascorbic acid solution for being 100mM by 40 μ L molar concentrations is added drop-wise to the chlorination that 1000 μ L molar concentrations are 1.5mM
It in palladium solution, is uniformly mixed rapidly, until color becomes light green color to get to Pd nano particle solution, is placed in storage at normal temperature, it is standby
With；Obtained Pd nano particle solution is mixed with the betulic acid nano liposomes that glutathione is modified by the volume ratio of 1:5
Uniformly, it is placed in shaking table, 30 DEG C, 115rpm, is protected from light and is incubated for 16 hours to get the betulic acid nanometer rouge coated to Pd nano particle
Plastid；
The betulic acid nano liposomes for taking 1500 μ L Pd nano particles to coat, it is 8mM's that 1000 μ L molar concentrations, which are added,
Chlorogold solution mixes well；Then the hydroxylamine hydrochloride solution that 500 μ L molar concentrations are 100mM will be added into mixed liquor, fills
Divide and be uniformly mixed, places it in shaking table, 30 DEG C, 115rpm, be incubated for 12 hours to get the white birch coated to nano flower-like palladium-gold
Resin acid liposome.
Nano flower-like palladium-gold cladding betulic acid liposome is characterized using ultraviolet-visual spectrometer, such as Fig. 3 institute
Show, it can be seen that after forming nano flower-like palladium-billon shell, exist in 500-900nm wide wave-length coverage close
Infrared absorption；Meanwhile nano flower-like palladium-gold cladding betulic acid liposome photothermal conversion 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 have good photothermal conversion performance.
Claims (1)
1. a kind of preparation method of nano flower-like palladium-gold cladding betulic acid liposome, the nano flower-like palladium-gold cladding
Betulic acid liposome is a kind of in betulic acid nano liposomes surface formation nano flower-like palladium-billon shell nanometer rouge
Plastid, particle size 80-100nm, near infrared absorption wavelength are 500-900nm, it is characterised in that: it includes following step
It is rapid:
(1) the betulic acid nano liposomes of glutathione modification are prepared
1. the mass ratio of each raw material are as follows: soybean lecithin: cholesterol: betulic acid=12~20:1.3~2.1:1, by every milli
Rising dehydrated alcohol and the summation of above-mentioned three kinds of Solute mass is added is 14.3~23.1mg, and dehydrated alcohol and ultrasound, ultrasonic function is added
Rate is 40~80 W, and ultrasonic time 10-20min makes it completely dissolved to obtain lipid soln；
2. glutathione is taken to be dissolved in deionized water, glutathione solution is obtained, contains 0.3~1mg in every ml deionized water
Glutathione；Volume ratio with lipid soln and glutathione solution is 1:1.5~3.5, and magnetic agitation and temperature, which control, to exist
38~47 DEG C, in the glutathione solution that 1. lipid soln that step is obtained is added dropwise to, obtain Liposomal suspensions；
3. 2. Liposomal suspensions that step is obtained are removed in 28~35 DEG C, 100~130rpm, 3~7 hours progress magnetic agitations
The dehydrated alcohol in Liposomal suspensions is removed, the betulic acid nano liposomes of glutathione modification are obtained；
(2) nano flower-like palladium-billon cladding betulic acid nano liposomes are prepared
1. be the ratio of 100:1~4 with the volume ratio of palladium chloride solution and ascorbic acid solution, by molar concentration be 50~
The ascorbic acid solution of 100mM is added drop-wise in the palladium chloride solution that molar concentration is 0.5~1.5mM, is uniformly mixed rapidly, directly
Become light green color to color to get to Pd nano particle solution, is placed in storage at normal temperature, it is spare；
2. by step 1. obtained in the betulic acid nanometer of glutathione modification that obtains of Pd nano particle solution and step (1)
Liposome is uniformly mixed by the volume ratio of 1:1~5, is placed in shaking table, 25~30 DEG C, it is small to be protected from light incubation 8~16 by 85~115rpm
When to get to Pd nano particle cladding betulic acid nano liposomes；
3. with the ratio of volume ratio 10:5~15 of chlorogold solution and the betulic acid nano liposomes of Pd nano particle cladding,
It is 2~8mM chlorauride that molar concentration is added in the betulic acid nano liposomes coated to 2. Pd nano particle that step obtains
Solution mixes well, and is 50~100mM hydroxylamine hydrochloride solution then to molar concentration is added in mixed liquor, is sufficiently mixed uniformly,
And the volume ratio of chlorogold solution and hydroxylamine hydrochloride solution is 10:1~5, places it in shaking table, 25~30 DEG C, 85~
115rpm is incubated for 6~12 hours to get the betulic acid liposome coated to nano flower-like palladium-gold.
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| CN104984339A (en) * | 2015-06-26 | 2015-10-21 | 燕山大学 | Oleanolic acid liposome coated with nanogold spherical shell and preparation method thereof |
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