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CN-122006666-A - Preparation method and application of traditional Chinese medicine component magnetic nanospheres

CN122006666ACN 122006666 ACN122006666 ACN 122006666ACN-122006666-A

Abstract

The invention discloses a preparation method and application of a traditional Chinese medicine component magnetic nanosphere; the magnetic microsphere takes Fe 3 O 4 magnetic nanospheres as cores, oleic acid is used for modifying the Fe 3 O 4 magnetic nanospheres, the fishing hook taking NDBA as a group and the traditional Chinese medicine active ingredient as bait are formed, the preparation method of the traditional Chinese medicine component magnetic nanospheres is simple to operate and mild in condition, and can enrich various components in the traditional Chinese medicine; the target is peeled off from the surface of the microsphere through specific elution, and high-resolution mass spectrometry identification and analysis are carried out, so that the target corresponding to the chemical components of the traditional Chinese medicine is obtained, and the direct acting target for researching the traditional Chinese medicine complex system is discovered.

Inventors

  • YIN XIAOYING
  • SHI DAMING
  • LU YANG
  • YAN YINAN
  • WANG HAN

Assignees

  • 上海工程技术大学

Dates

Publication Date
20260512
Application Date
20260217

Claims (10)

  1. 1. The preparation method of the magnetic nanospheres with traditional Chinese medicine components is characterized by comprising the following steps: The surface of the traditional Chinese medicine component magnetic nanosphere is modified by (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) to obtain the Fe 3 O 4 magnetic nanosphere with azo groups and carboxyl functional groups, and the method comprises the following steps: Step 1, preparation of (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid), designated NDBA; step 2, preparing (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) modified Fe 3 O 4 -OA magnetic nanospheres, which are marked as Fe 3 O 4 -OA-NDBA.
  2. 2. The method for preparing the magnetic nanospheres with traditional Chinese medicine components according to claim 1, wherein the preparation of the (E) -4,4'- (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) in the step 1 is characterized in that 4-nitro-3- (trifluoromethyl) benzoic acid is dissolved in 10 times of sodium hydroxide solution (17%), the temperature is 50-55 ℃, preferably 50 ℃, after the temperature is reached, 5 times of saturated glucose aqueous solution is dropwise added for reaction for 0.8-2 hours, the mixed solution obtained by the reaction is subjected to acidification and precipitation by acetic acid, the pH value of the acidification process is 5.8-6.2, solid-liquid separation is carried out after precipitation, and the (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid is obtained after drying.
  3. 3. The method for preparing the magnetic nanospheres with Chinese medicinal components according to claim 2, wherein after the temperature is reached, a saturated glucose aqueous solution with 5 times molar quantity is dripped for reaction for 1.5 hours.
  4. 4. The method for preparing the magnetic nanospheres with Chinese medicinal components according to claim 2, wherein acetic acid is used in the acidification precipitation process.
  5. 5. The method for preparing the magnetic nanospheres with the traditional Chinese medicine components according to claim 1, wherein the preparation of the (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) modified Fe 3 O 4 -OA magnetic nanospheres in step 2 is characterized by taking the preparation of the Fe 3 O 4 -OA nanospheres in EDC, HCl and NHS mixed solution (M EDC·HCl ：M NHS =1:1,H 2 O), stirring at room temperature for 2 hours, magnetically separating the product, washing with absolute ethyl alcohol, adding the separated product into the (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) solution (10 mmol, absolute ethyl alcohol) in step 1, stirring at room temperature for overnight, washing with absolute ethyl alcohol, magnetically separating the product, putting the product into ethanolamine for sealing for 1 hour, magnetically separating the product, and drying to obtain the Fe 3 O 4 -OA-NDBA-OA magnetic nanospheres with the (E) -4 ' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) modified Fe 3 O 4 - 3 O 4 -OA.
  6. 6. The method for preparing the traditional Chinese medicine component magnetic nanospheres according to claim 5, wherein the application method comprises the steps of adding the magnetic nanomaterial into a small-sized grinding bottle containing traditional Chinese medicine liquid, placing the small-sized grinding bottle into a shaking table for incubation, adding a magnetic field after incubation is completed to separate the magnetic nanomaterial enriched with the traditional Chinese medicine component from the traditional Chinese medicine liquid, detecting peak areas of the traditional Chinese medicine liquid before and after enrichment by using a high performance liquid chromatograph, and calculating the enrichment rate of the magnetic nanomaterial on the traditional Chinese medicine component.
  7. 7. The method for preparing the magnetic nanospheres with Chinese medicinal components according to claim 6, wherein the incubation temperature is 30 ℃.
  8. 8. The method for preparing the magnetic nanospheres with Chinese medicinal components according to claim 6, wherein the incubation time is 8-12h.
  9. 9. The use of the magnetic nanospheres enriched in herbal components as claimed in claim 6 for the discovery of target targets in medicine.
  10. 10. An application of the magnetic nanospheres with traditional Chinese medicine components prepared by the preparation method of the magnetic nanospheres with traditional Chinese medicine components according to any one of claims 1-9 in research and discovery of traditional Chinese medicine targets.

Description

Preparation method and application of traditional Chinese medicine component magnetic nanospheres Technical Field The invention relates to the technical field of nano material preparation and application, in particular to a preparation method and application of a composite material capable of enriching traditional Chinese medicine components. Background The traditional Chinese medicine is an important treasure in Chinese medicine, and is widely applied to clinic because of the effects of multiple components, multiple targets and multiple layers. However, the action targets and action mechanisms of many traditional Chinese medicines and active ingredients thereof are not clear, so that the application of the traditional Chinese medicines in clinic is limited. Currently, the main methods of drug target research can be broadly divided into two categories, experimental verification techniques based on chemistry and bioscience, and computational methods including virtual screening and predictive modeling. Experimental verification techniques in chemistry and biology include methods such as solid phase carrier microsphere capture and quantitative proteomics techniques, including SILAC, CETSA, SPROX and DARTS. In contrast, computing virtual screening strategies includes network pharmacology and molecular docking screening. By comparing the different technical characteristics, a target spot discovery related method suitable for a complex traditional Chinese medicine system is also lacking. The ferroferric oxide (Fe 3O4) magnetic nano-particles have attracted wide attention in the fields of biotechnology and medicine research due to the unique physicochemical properties. In particular, such nanoparticles generally exhibit good superparamagnetism, lower biotoxicity, higher specific surface area, and good surface modification. Based on these characteristics, the Fe 3O4 magnetic nanoparticles can be covalently or non-covalently bound to a variety of bioactive molecules through surface functionalization (e.g., modification of functional groups such as carboxyl, amino, epoxy, etc.), thereby achieving immobilization of these molecules. The types of molecules that can be immobilized are wide ranging and include biological macromolecules such as proteins, polypeptides, enzymes, antibodies, and chemical small molecules (e.g., active ingredients in traditional Chinese medicine). The material has the potential of being applied to enrichment of traditional Chinese medicine components, and how to effectively fix the traditional Chinese medicine components with various structures is a technology to be developed urgently. Therefore, a magnetic nanoparticle which can be used for fixing active ingredients of traditional Chinese medicines efficiently and stably is needed, the enrichment capability of the magnetic nanoparticle can be improved, small active molecules of different types of traditional Chinese medicines can be fixed, good biocompatibility and magnetic responsiveness can be kept, and the magnetic nanoparticle has important significance in promoting the discovery of target points of traditional Chinese medicines and clinical application and research. Disclosure of Invention Aiming at the problems existing in the prior art, the invention provides a composite magnetic nano material for carrying out functional modification on the surface of Fe 3O4 nanospheres, so that the composite magnetic nano material can effectively enrich active ingredients of traditional Chinese medicines and identify the action targets of the enriched traditional Chinese medicines. The invention discloses a preparation method of a functional modified Fe 3O4 magnetic nanosphere, which uses Fe 3O4 nanospheres as a modified core carrier, oleic Acid (OA) as a biological performance improver to improve biocompatibility, and (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) (NDBA) as an active ingredient of a traditional Chinese medicine covalently coupled with a molecular fishhook. Further, the preparation method of the functional modified Fe 3O4 magnetic nanospheres (Fe 3O4 -OA-NDBA) comprises the following steps: Step 1, preparing Fe 3O4 magnetic nanospheres; Step 2, preparing an oleic acid modified Fe 3O4 magnetic nanosphere, namely Fe 3O4 -OA; Step 3, preparation of (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid), designated NDBA; Step 4, preparing (E) -4,4' - (diene-1, 2-diyl) bis (3- (trifluoromethyl) benzoic acid) modified Fe 3O4 -OA magnetic nanospheres, which are marked as Fe 3O4 -OA-NDBA; Step 5, preparing Fe 3O4 -OA-NDBA magnetic nanospheres enriched with traditional Chinese medicine components; Further, the Fe 3O4 magnetic nanospheres prepared in the step 1 are prepared by adding deionized water into a round-bottom flask with the addition amount of 50-150ml, simultaneously adding FeCl 3·6H2 O and FeCl 2·4H2 O, wherein the molar ratio of FeCl 3·6H2 O to FeCl 2·4H2 O is 2:1, protecting by nitrogen (N 2), m