Silk fibroin nanoparticle and application thereof

A nanoparticle and silk protein technology, applied in the field of medicine, can solve the problems of toxicity, inability to apply TPL and CL, poor water solubility, etc., achieve high safety, inhibit cancer cell proliferation, and strengthen the effect of inhibition

Active Publication Date: 2017-08-15
JIAXING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to their poor water solubility and severe toxic

Method used

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  • Silk fibroin nanoparticle and application thereof
  • Silk fibroin nanoparticle and application thereof
  • Silk fibroin nanoparticle and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of silk protein nanoparticles

[0043] 1. Preparation, purification and molecular weight of regenerated silk fibroin (SF)

[0044] Cut the purchased silkworm cocoons into small pieces, and in 0.02M Na 2 CO 3 Degumming twice for 30 minutes each, and rinsed thoroughly with deionized water. After air drying, the degummed silk fibroin (SF) was dissolved in the ternary system CaCl 2 -CH 3 CH 2 OH-H 2 O (1:2:8 molar ratio), stirring was continued at 75°C for 4 hours. In addition, the resulting SF solution was centrifuged at 4500 rpm for 5 minutes. The supernatant was carefully collected and dialyzed against deionized water for 3 days in a Viskase dialysis membrane (MWCO 7000 Da) to remove salts and ethanol. After centrifugation at 14000 rpm for 15 minutes, the concentration of the prepared silk fibroin solution was measured using a NanoDrop™ 2000 / 2000c spectrophotometer. The molecular weight range of the extracted SF was analyzed by SDS-PAGE (8%...

Embodiment 2

[0058] Example 2 Nanoparticle Characterization

[0059] 1. Method

[0060] 1.1 Particle Size, Zeta Potential and Morphology

[0061] Freeze-dried SFNPs were dispersed in deionized water (pH 7.0). The average size and zeta potential of NPs (Blank-SFNP, TPL-SFNP, CL-SFNP) were determined by dynamic light scattering detection (NanobrookOmni, Brookhaven Instrument Corp) (all measurements were performed three times at room temperature). Morphological examination of NPs and SFs was performed by transmission electron microscopy (TEM, H600, HITACHI, Japan). SF was diluted and lyophilized NPs were resuspended with deionized water. Additional samples were prepared by placing a drop of the dilute suspension on a copper grid coated with a carbon film. The surface morphology of the samples was then observed under TEM.

[0062] 1.2 Infrared spectrum IR absorption and β-sheet content

[0063] FTIR spectra of drug-loaded SFNP as well as free drug were performed using a Fourier transform...

Embodiment 3

[0083] Example 3 Safety Evaluation of Blank SFNPs and Drug-loaded SFNPs

[0084] 1. In Vitro Hemolytic Assay

[0085] The hemolytic assay of SFNP and drug-loaded SFNPs was carried out in vitro to investigate their biocompatibility. Briefly, 0.2 mL of 3.8% sodium citrate was mixed with 4 mL of fresh mouse blood, then centrifuged at 3,000 rpm for 10 min, the precipitate was collected and suspended in 37°C PBS (pH 7.4) and washed 3 times to obtain 5% (v / v) suspension of RCB. 10 mg / mL suspended NPs (Blank-SFNPs, TPL-SFNPs and CL-SFNPs) were incubated with RBCs in PBS. 0.1% Triton X-100 and PBS were added to the RBCs solution as positive and negative controls, respectively. All samples were incubated for 2 hours at room temperature. Then, the sample was centrifuged for 5 min, and 100 μL of supernatant was collected carefully, and transferred to a 96-well plate, and the content of hemoglobin was detected with a microplate reader. Calculate the percent hemolysis according to the...

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Abstract

The invention discloses a silk fibroin nanoparticle. The silk fibroin nanoparticle contains silk fibroin and an active drug, wherein the silk fibroin loads the active drug, and the active drug is selected from triptolide and tripterine. The invention also discloses a preparation method and application of the silk fibroin nanoparticle. The silk fibroin nanoparticle improves the water solubility of the triptolide and tripterine and can resist to proliferation of cancer cells more effectively; and meanwhile, combined utilization of a silk fibroin nanoparticle loading triptolide (TPL-SFNPs) and a silk fibroin nanoparticle loading tripterine (CL-SFNPs) can synergistically inhibit the growth of the cancer cells, and effect is significant, so that the silk fibroin nanoparticle has good application prospects in treatment of cancers, especially pancreatic cancer.

Description

technical field [0001] The invention relates to the technical field of medicine, in particular to a silk protein nanoparticle loaded with triptolide (TPL) or triptolide (CL) and an application thereof. Background technique [0002] Pancreatic cancer (PC) is one of the most aggressive human malignancies, with a 5-year survival rate of only 8%, which has a relatively low incidence of 2% in all diagnosed cancers, but accounts for 6% of cancer deaths. Chemotherapy for PC has strong toxic side effects, and PC has potential drug resistance, both of which lead to the unfortunate death of patients in many cases. The current standard chemotherapy drug for pancreatic cancer is gemcitabine, however, its efficacy is far from satisfactory, one reason being due to the complex tumor microenvironment that reduces effective drug delivery to target cancer cells. To date, no drug has been shown to significantly prolong clinical overall survival. Therefore, new therapeutic strategies for pan...

Claims

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Application Information

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IPC IPC(8): A61K9/51A61K47/42A61K31/585A61K31/56A61P35/00
CPCA61K9/5169A61K31/56A61K31/585A61K2300/00
Inventor 丁宝月王竞华
Owner JIAXING UNIV
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