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CN-121991260-A - Lentinan-gold nanocluster fluorescent probe and preparation method and application thereof

CN121991260ACN 121991260 ACN121991260 ACN 121991260ACN-121991260-A

Abstract

The invention discloses a lentinan-gold nanocluster fluorescent probe and a preparation method and application thereof, and relates to the technical field of biomedical materials and natural polysaccharide functionalization. According to the invention, the gold nanoclusters modified by the aminated lentinan and the glutathione are used as raw materials, and are subjected to covalent coupling under mild conditions to obtain the lentinan-gold nanocluster fluorescent probe. The preparation process avoids strong acid, strong alkali and high temperature conditions, and can effectively retain natural triple helix conformation of lentinan, thereby maintaining inherent antitumor bioactivity. The prepared fluorescent probe has uniform particle size and average size of about 150 nm, and is beneficial to realizing tumor part enrichment through the enhanced permeation and retention effect (EPR effect) of tumors. In the tumor microenvironment, the inherent anti-tumor activity of lentinan and the tumor cytotoxicity induced by the peroxidase-like activity shown by Au-GSH generate a synergistic effect, and the probe has stable fluorescent signals, so that in-vivo tracing and imaging can be realized.

Inventors

  • WAN HAO
  • FENG LEI
  • LU XINJIAN

Assignees

  • 南昌大学

Dates

Publication Date
20260508
Application Date
20260129

Claims (10)

  1. 1. The preparation method of the lentinan-gold nanocluster fluorescent probe is characterized by comprising the following steps of: the method comprises the steps of taking aminated lentinan and glutathione modified gold nanoclusters as raw materials, and carrying out covalent coupling on the raw materials under mild conditions to prepare the lentinan-gold nanocluster fluorescent probe.
  2. 2. The preparation method of claim 1, wherein the mass ratio of the aminated lentinan to the glutathione-modified gold nanoclusters is 0.8-1.2:0.8-1.2.
  3. 3. The preparation method according to claim 1, wherein the preparation method of the aminated lentinan comprises the following steps: Dissolving lentinan in an organic solvent to obtain a lentinan solution, and adjusting the pH value of the lentinan solution to 8.5-9; Adding the succinic anhydride solution into the lentinan solution for reaction, keeping the pH value of the system to be 8.5-9 in the reaction process, adjusting the pH value of the system to be 6.5-7 after the reaction is finished, and dialyzing, concentrating, precipitating, washing and drying the product to obtain LNT-COOH; Dissolving LNT-COOH in an organic solvent, adding HATU, HOBT and DIEA for reaction, adding H 2 N-NHBOC for amination reaction, adding trifluoroacetic acid for stirring after the reaction is finished, dialyzing, concentrating and drying the product to obtain the aminated lentinan.
  4. 4. The preparation method of the glutathione-modified gold nanoclusters according to claim 1, comprising the steps of: HAuCl 4 H 2 O is dissolved in water, glutathione is added to generate microemulsion suspension, naBH 4 solution is added rapidly after intense stirring, stirring is continued, and the product is filtered, purified and concentrated to obtain the gold nanocluster modified by glutathione.
  5. 5. The preparation method according to claim 1, characterized in that it comprises the following specific steps: dissolving glutathione modified gold nanoclusters in dimethyl sulfoxide to obtain an Au-GSH solution, sequentially adding a condensing agent, an auxiliary agent and an alkaline catalyst into the Au-GSH solution, stirring and activating, adding aminated lentinan for reaction, and performing dialysis, concentration and freeze-drying treatment after the reaction is finished to obtain the lentinan-gold nanocluster fluorescent probe.
  6. 6. The preparation method of claim 5, wherein the concentration of the Au-GSH solution is 1.5-2.5 mg/mL, and the molar ratio of the condensing agent, the auxiliary agent and the alkaline catalyst is 0.1:0.1:0.2.
  7. 7. The method according to claim 5, wherein, The condensing agent is HATU, the auxiliary agent is HOBT, and the alkaline catalyst is DIEA; The dialysis adopts a dialysis bag with a molecular weight cut-off value of 50000 Da; the reaction time of adding the aminated lentinan is 1.5-2.5 h.
  8. 8. A lentinan-gold nanocluster fluorescent probe, characterized in that it is obtained by the preparation method according to any one of claims 1 to 7.
  9. 9. The lentinan-gold nanocluster fluorescent probe according to claim 8, wherein the average particle size of the lentinan-gold nanocluster fluorescent probe is 140-160 nm, and the lentinan-gold nanocluster fluorescent probe retains a triple helix structure of lentinan.
  10. 10. The use of a lentinan-gold nanocluster fluorescent probe according to any one of claims 8-9 in the preparation of tumor imaging products or tumor treatment products or tumor diagnosis and treatment integrated products.

Description

Lentinan-gold nanocluster fluorescent probe and preparation method and application thereof Technical Field The invention relates to the technical field of biomedical materials and natural polysaccharide functionalization, in particular to a lentinan-gold nanocluster fluorescent probe and a preparation method and application thereof. Background Lentinan (LNT) is a natural beta-glucan derived from lentinus edodes, has definite biological activities such as immunoregulation, anti-tumor and anti-inflammatory, and has been widely studied and partially applied to clinical adjuvant therapy. However, lentinan lacks imaging capability, is difficult to realize visual monitoring of in vivo distribution, metabolism and targeted enrichment process, and limits further application of lentinan in the fields of precise medicine and tumor diagnosis and treatment integration. Studies have shown that lentinan is highly dependent on its natural triple helix structure, which is an important structural basis for its immune enhancing and antitumor effects. However, existing polysaccharide modification or nanocrystallization methods often require relatively harsh reaction conditions, such as strong acid, strong base or high temperature treatment, which are prone to break the triple helix conformation of the polysaccharide, resulting in a significant decrease in biological activity. Therefore, how to effectively retain the triple helix structure of lentinan while achieving functional modification is a key technical problem to be solved in the field. The gold nanocluster has excellent fluorescence performance, biocompatibility and peroxidase-like activity, and has good application prospect in the fields of biological imaging and tumor treatment. Especially glutathione modified gold nanoclusters (Au-GSH), not only have stable fluorescence signals, but also can catalyze active oxygen generation in tumor microenvironment, thereby inducing tumor cell injury. However, au-GSH has a particle size of only about 2nm, lacks tumor enrichment capability, and has limited in vivo enrichment efficiency. Therefore, a novel preparation method which can effectively combine Au-GSH with lentinan under mild conditions, not only maintains the triple helix structure and biological activity of the lentinan, but also endows the lentinan with fluorescence tracing and synergistic anti-tumor functions is developed, and has important significance for promoting the application of natural polysaccharide in tumor diagnosis and treatment integration. Disclosure of Invention The invention aims at least solving one of the technical problems in the prior art, provides a lentinan-gold nanocluster fluorescent probe and a preparation method and application thereof, and in particular relates to a preparation method of a lentinan-gold nanocluster fluorescent probe which takes lentinan as a main body and gold nanoclusters as fluorescence and catalysis units, which is suitable for biomedical applications such as tumor imaging, tumor treatment and diagnosis and treatment integration. The invention provides a preparation method of a lentinan-gold nanocluster fluorescent probe, which is used for preparing an LNT-Au fluorescent probe by combining gold nanoclusters Au-GSH with LNT-NH 2. The method can effectively retain triple helix structure and bioactivity of lentinan, and simultaneously endow probe with fluorescent tracing function and tumor enrichment capability, and has wide tumor diagnosis and treatment application prospect. The technical scheme of the invention is as follows: In a first aspect, the invention provides a preparation method of a lentinan-gold nanocluster fluorescent probe, which comprises the following steps: the method comprises the steps of taking aminated lentinan and glutathione modified gold nanoclusters as raw materials, and carrying out covalent coupling on the raw materials under mild conditions to prepare the lentinan-gold nanocluster fluorescent probe. According to one specific embodiment of the invention, the mass ratio of the aminated lentinan to the glutathione modified gold nanoclusters is 0.8-1.2:0.8-1.2. In one embodiment of the present invention, the preparation method of the aminated lentinan comprises the following steps: Dissolving lentinan in an organic solvent to obtain a lentinan solution, and adjusting the pH value of the lentinan solution to 8.5-9; Adding the succinic anhydride solution into the lentinan solution for reaction, keeping the pH value of the system to be 8.5-9 in the reaction process, adjusting the pH value of the system to be 6.5-7 after the reaction is finished, and dialyzing, concentrating, precipitating, washing and drying the product to obtain LNT-COOH; Dissolving LNT-COOH in an organic solvent, adding HATU, HOBT and DIEA for reaction, adding H 2 N-NHBOC for amination reaction, adding trifluoroacetic acid for stirring after the reaction is finished, dialyzing, concentrating and drying the product to obtain the aminate