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Bufalin (BUF)-carrying nano liposome modified by hyaluronic acid as well as preparation method and application of BUF-carrying nano liposome

A technology of hyaluronic acid modification and nano-liposome, which is applied in the field of medicine to achieve the effects of prolonging the efficacy, increasing the accumulation and increasing the sensitivity

Inactive Publication Date: 2020-10-16
SHANGHAI PUTUO DISTRICT CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] There is no report about a hyaluronic acid-modified bufatoxin-loaded nanoliposome of the present invention and its preparation method and application

Method used

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  • Bufalin (BUF)-carrying nano liposome modified by hyaluronic acid as well as preparation method and application of BUF-carrying nano liposome
  • Bufalin (BUF)-carrying nano liposome modified by hyaluronic acid as well as preparation method and application of BUF-carrying nano liposome
  • Bufalin (BUF)-carrying nano liposome modified by hyaluronic acid as well as preparation method and application of BUF-carrying nano liposome

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The preparation of embodiment 1 HA-DOPE

[0037] Hyaluronic acid (HA) activation solution was prepared by 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS), containing phosphatidyl The suspension of ethanolamine (DOPE) is added to the activation solution to activate the reaction, and the unreacted DOPE, EDC and other reagents are removed by dialysis to prepare HA-DOPE. The HA-DOPE freeze-dried powder and KCl powder were mixed and ground at a ratio of 1:100, sieved to make the particle size below 2.5 μm, and put into a tablet former. Pressurized (5-10t / cm 2 ) in about 3 minutes to obtain a transparent sheet, which can be tested on the machine. The infrared spectra of HA, DOPE and HA-DOPE are shown in the figure, in A 1690~1650cm -1 The multiplet is the stretching vibration peak of C=O bond and amide in HA, 2910cm in B -1 , 1780cm -1 , 1380.97cm -1 It is the characteristic absorption peak of long-chain methylene in DOPE, 1735.44 cm ...

Embodiment 2

[0038] Example 2 Preparation of HA / lip-BUF

[0039] Precisely weigh 72 mg of S100 and 24 mg of cholesterol in a round bottom flask, add 10 mL of chloroform and a certain amount of BUF (and HA-DOPE), stir for 10 min to form a uniform solution, remove the chloroform by rotary evaporation in a water bath at 30-60 ° C, and form a uniform transparent film. Then add 9 mL of chloroform and 3 mL of PBS, shake vigorously to mix, and sonicate for 5 min until a stable W / O phase is formed. Rotary steam in a water bath at 30-60°C until it becomes flocculent, then add 5 mL of PBS and continue rotating until the hydration is complete (hydration for 1 hour), and the bufalin liposome is obtained. Probe ultrasound for 10min, 10-30%W, 5-10s on, 5s off, standby. After the prepared liposomes were diluted to an appropriate concentration with distilled water, the particle diameter and PDI of the liposomes were measured with a Malvern Zetasizer particle size analyzer. The morphology and size of the...

Embodiment 3

[0040] Embodiment 3 measures the encapsulation efficiency of loading BUF liposome

[0041] Column: C18 VanGuard Pre-column, 1.7μm, 2.1mm X 5mm; mobile phase - acetonitrile: water = 4: 6; flow rate: 0.5mL / min; column temperature: 10-40°C; detection wavelength: 280nm; injection volume: 2μL. The encapsulation efficiency of BUF-loaded liposomes was determined by ultrafiltration centrifugation.

[0042] First, take 300 μL of liposomes and add 4 The upper part of the ultrafiltration centrifuge tube of u, 2000-5000r·min -1 Centrifuge for 30-40min, so that the liposomes are trapped on the upper part of the ultrafiltration tube, while the unencapsulated free drug is centrifuged to the bottom of the ultrafiltration tube. Take 100 μL of the upper liquid and add 900 μL of methanol, ultrasonically break the emulsion, centrifuge to get the supernatant, and use it for measurement. The result is as image 3 As shown, the standard curve was drawn with the concentration of bufalin as the ...

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Abstract

The invention relates to the technical field of medicine, in particular to a bufalin (BUF)-carrying nano liposome modified by hyaluronic acid as well as a preparation method and application of the BUF-carrying nano liposome. A film dispersion method is combined with an ultrasonic method to prepare the liposome with a smaller particle size and dispersion uniformity, the hyaluronic acid is linked tothe DOPE phospholipid through chemical modification, and a dialysis bag method is used for purification. The solubility of the BUF can be improved, the tumor targeting effect can be enhanced, toxic and side effects can be reduced, the drug resistance of breast cancer cells to docetaxel (DCT) can be reversed, and new dosage form application is provided for the clinical application of the BUF.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a hyaluronic acid-modified nano-liposome loaded with bufatoxin and its preparation method and application. Background technique [0002] Bufadienolide (BUF) is a kind of active bufadiene lactone extracted from the traditional Chinese medicine Bufa venom, which has a wide range of anticancer effects. Ye M, Guo H, Guo H, et al. (2006) Simultaneous determination of cytotoxic bufadienolides in the Chinese medicine ChanSu by high-performance liquid chromatography coupled with photodiode array and mass spectrometry detections. J Chromatogr B 838(2):86–95. However, due to the low solubility of BUF, weak sensitivity to diseases and poor selectivity to normal tissues and organs, it is easy to cause vascular irritation, anaphylactic shock, fever and sinus bradycardia. These shortcomings make it a great challenge for clinical application. Therefore, improving the solubility and targeting...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K47/36A61K31/585A61P35/00A61P15/14
CPCA61K9/1272A61K9/1277A61K47/36A61K31/585A61P35/00A61P15/14
Inventor 袁易张蝶周霖曾海荣郭腾王之颖董君君王旭慧
Owner SHANGHAI PUTUO DISTRICT CENT HOSPITAL
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