Radix rehmanniae protein nano-particles and preparation method thereof

A technology of nanoparticles and rehmannia, which is applied in the field of new dosage forms and preparations of protein drugs, can solve the problems that rehmannia nanoparticles have not been reported in the literature, and achieve the effects of easy and stable storage, high safety and good resolubility

Active Publication Date: 2017-12-01
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, the research on rehmannia glutinin nanoparticles has not been reported in the literature at home and abroad.

Method used

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  • Radix rehmanniae protein nano-particles and preparation method thereof
  • Radix rehmanniae protein nano-particles and preparation method thereof
  • Radix rehmanniae protein nano-particles and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Extraction of Rehmannia Protein DHP1

[0032] Fresh Rehmannia glutinosa was ground by a juicer, the ratio of solid to liquid was 1:3, 0.2 mol / L, pH 7.0 phosphate buffer was added to mix evenly, placed at 4°C, stirred and extracted for 24 h; Extract the liquid, discard the filter residue, centrifuge the filtrate at 15000 rpm for 20 min at 4 °C, and collect the supernatant; under the condition of magnetic stirring at 4 °C, slowly add anhydrous ammonium sulfate to the extract of rehmannia glutinosa to make the solution salt. The concentration reached 20%. After it was completely dissolved, it was allowed to stand at 4 °C for 4 h, centrifuged at 12,000 rpm for 20 min, and the supernatant was collected; continue to slowly add anhydrous ammonium sulfate to the collected supernatant until the saturation was 40%. After complete dissolution, collect the supernatant in the same steps as before; continue to add ammonium sulfate to the supernatant until the saturation is...

Embodiment 2

[0035] Example 2: Extraction of Rehmantin DHP2

[0036] Rehmannia glutinosa was ground by a high-speed pulverizer, and phosphate buffer solution of 0.2 mol / L and pH 7.0 was added at a ratio of 1:3 to mix evenly, placed at 4 °C, and stirred for leaching for 24 h; Extract the liquid, discard the filter residue, centrifuge the filtrate at 15000 rpm for 20 min at 4 °C, and collect the supernatant; under the condition of magnetic stirring at 4 °C, slowly add anhydrous ammonium sulfate to the extract of rehmannia glutinosa to make the solution salt. The concentration reached 20%. After it was completely dissolved, it was left standing at 4 °C for 4 h, centrifuged at 12,000 rpm for 20 min, and the supernatant was collected; continue to slowly add anhydrous ammonium sulfate to the collected supernatant until the saturation was 40%. After complete dissolution, collect the supernatant in the same steps as before; continue to add ammonium sulfate to the supernatant until the saturation i...

Embodiment 3

[0039] Example 3: Preparation of retinoid nanoparticles

[0040] The retinoid was prepared into a 1 mg / mL pH 6.5 salt solution, heated at 70° C. for 1 hour, and the unreacted retinoid was removed by TSK gelG6000PW gel chromatography to obtain the retinoid nanoparticles. The properties of retinoid nanoparticles were studied by dynamic light scattering, multi-angle laser light scattering and scanning electron microscopy. The electron microscope observation map of retinoids nanoparticles is attached Figure 9 .

[0041] The particle size and surface potential were measured with a laser particle size analyzer, and the average particle size was measured to be 120.61 ± 0.46 nm, and the surface charge was negative, ranging from -18 ± 5mV. The size distribution of rehmantin nanoparticles is shown in the attached Figure 10 .

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Abstract

The invention discloses radix rehmanniae proteins and a preparation method of radix rehmanniae protein nano-particles. Each particle is composed of two correspondingly proteins, wherein the N-end primary structural sequences of the two radix rehmanniae proteins are SEQ ID NO:1: R-E-K-D-I-A-G-A-V-Q-T-V and SEQ ID NO:2: R-E-K-D-I-V-X correspondingly. The radix rehmanniae proteins are extracted and separated from fresh radix rehmanniae, or raw radix rehmanniae or prepared radix rehmanniae and stably exist in the radix rehmanniae raw materials which are used traditionally. No cross-linking agent is involved in the preparation method of the radix rehmanniae protein nano-particles, finished products of the radix rehmanniae proteins are widely used in various decoctions prepared from the raw materials of the radix rehmanniae through a traditional hot processing technique, wherein the decoctions are taken by a large number of people for many years and are high in safety. The radix rehmanniae protein nano-particles are safe and efficient, the efficacy of the radix rehmanniae protein nano-particles is achieved safely and efficiently, and in addition, the radix rehmanniae protein nano-particles are expected to be combined with other raw materials, so that the effects are achieved efficiently and collaboratively.

Description

technical field [0001] The invention relates to a new dosage form and preparation technology of protein medicines in the technical field of medicine, in particular to a rehmantin nanoparticle and a preparation method thereof. Background technique [0002] In recent decades, nanoparticle research has been extensive. After nanomaterialization, the physicochemical properties (optical, electrical, surface activity, etc.) of nanomaterials will show completely different properties from the original materials, with targeting and penetrating properties. At the same time, the disadvantages of biocompatibility, cytotoxicity and degradability are also exposed. At present, there is no natural research model for the biological macromolecules (such as serum albumin, silk protein, etc.) used in the research of nano-drug carriers. The physicochemical properties of nanoparticles and their related biological safety also require long-term observation. Comprehensive assessment. The potential ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K9/19A61P39/06C07K14/415C07K1/36C07K1/30C07K1/20C07K1/18C07K1/16G01N30/02A61K36/804
CPCA61K9/19A61K36/804A61K38/168C07K14/415G01N30/02
Inventor 丁伟柯李晶周建武饶平凡
Owner ZHEJIANG GONGSHANG UNIVERSITY
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