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CN-121971657-A - Polypeptide nano material for reducing tumor dryness by combining NRP1 as well as preparation method and application thereof

CN121971657ACN 121971657 ACN121971657 ACN 121971657ACN-121971657-A

Abstract

The invention relates to the technical field of nanomaterials, in particular to a polypeptide nanomaterial for reducing tumor dryness by combining NRP1, a preparation method and application thereof, wherein the polypeptide nanomaterial for reducing tumor dryness by combining NRP1 comprises a polypeptide with an amino acid sequence shown as SEQ ID NO. 1 and a hydrophobic fluorescent group connected with last lysine of the polypeptide through an amide bond, and the polypeptide comprises ligand peptide of NRP1 and a fiber deformation primitive connected with the ligand peptide through the amide bond. The polypeptide nano material is self-assembled in aqueous solution through hydrophilic and hydrophobic effects to form spherical nano particles, is combined with NRP1 on tumor cell membranes based on the amino acid sequence of targeted NRP1 as ligand peptide, and further induces the spherical nano particles to be deformed into a nanofiber network structure at tumor sites in situ through receptor-ligand interaction, and stays in tumor areas for a long time, so that NRP1 mediated passages are blocked completely for a long time, the characteristics of tumor stem cells are regulated down, and finally the treatment effect of tumors is enhanced.

Inventors

  • ZHANG LU
  • SHEN JUN
  • Zhao Jiaru
  • MAO CHUNPING

Assignees

  • 南方科技大学

Dates

Publication Date
20260505
Application Date
20251231

Claims (10)

  1. 1. A polypeptide nanomaterial for combining NRP1 to down regulate tumor dryness is characterized by comprising a polypeptide with an amino acid sequence shown as SEQ ID NO. 1 and a hydrophobic fluorescent group connected with last lysine of the polypeptide through an amide bond, wherein the polypeptide comprises a ligand peptide of NRP1 and a fiber deformation motif connected with the ligand peptide through an amide bond, and the hydrophobic fluorescent group is connected with one end of the fiber deformation motif, which is far away from the ligand peptide, through the last lysine.
  2. 2. The polypeptide nanomaterial for binding NRP1 to down-regulate tumor dryness according to claim 1, wherein the hydrophobic fluorescent group is selected from one of protoporphyrin, 7-nitrobenzodiazole, bipyrene molecule.
  3. 3. The polypeptide nanomaterial for binding NRP1 to down-regulate tumor dryness according to claim 1, wherein the polypeptide nanomaterial has a structural formula: 。
  4. 4. a method of preparing a polypeptide nanomaterial for down-regulating tumor dryness in combination with NRP1 as claimed in any one of claims 1 to 3, comprising the steps of: Swelling the chlorine resin to obtain a first product; Mixing K [ Fmoc-Lys (Boc) -OH ], N-diisopropylethylamine and N, N-dimethylformamide, and then reacting with the first product to obtain a second product; A mixed system of methanol, N-diisopropylethylamine and N, N-dimethylformamide is adopted to react with the second product, so as to obtain a third product; Coupling is sequentially carried out on the main chain of the third product according to the sequence of aspartic acid D, alanine A, lysine K (Boc), leucine L, cysteine C, lysine K (Boc), leucine L, valine V, phenylalanine F and lysine K (Dde) to obtain a fourth product; removing a main chain amino Fmoc protecting group in the fourth product, and using acetic anhydride to end-cap the amino exposed by the main chain to obtain a fifth product; removing a side chain amino Dde protecting group of lysine K (Dde) in the fifth product by using an N, N-dimethylformamide solution containing hydrazine hydrate to obtain a sixth product; Mixing a hydrophobic fluorescent group, an additive, N' -diisopropylcarbodiimide and N, N-dimethylformamide with the sixth product, and carrying out amidation reaction to obtain the nano polypeptide material.
  5. 5. The method of preparing a polypeptide nanomaterial for NRP 1-binding down-regulating tumor dryness according to claim 4, wherein in the step of mixing K [ Fmoc-Lys (Boc) -OH ], N-diisopropylethylamine and N, N-dimethylformamide and then reacting with the first product, the volume ratio of K [ Fmoc-Lys (Boc) -OH ], N-diisopropylethylamine and N, N-dimethylformamide is (2.5-3.5): 5-7): 1.
  6. 6. The method for preparing polypeptide nanomaterial for reducing tumor dryness in combination with NRP1 according to claim 4, wherein the volume ratio of methanol, N-diisopropylethylamine, N-dimethylformamide in the mixed system of methanol, N-diisopropylethylamine and N, N-dimethylformamide is (0.5-1.5): (1.5-2.5): (6-8).
  7. 7. The method for preparing a polypeptide nanomaterial for binding NRP1 to down-regulate tumor dryness according to claim 4, wherein in the step of mixing a hydrophobic fluorescent group, an additive, N '-diisopropylcarbodiimide, N-dimethylformamide and the sixth product, a mass ratio of the hydrophobic fluorescent group, the additive, the N, N' -diisopropylcarbodiimide and the chlorine resin is (2.5-3.5): (5.5-6.5): 1.
  8. 8. The method for preparing polypeptide nanomaterial for reducing tumor dryness in combination with NRP1 as claimed in claim 4, further comprising draining the chlorinated resin in the product after the amidation reaction, adding a cutting liquid, and performing rotary evaporation, precipitation, centrifugation and drying to obtain the polypeptide nanomaterial; The cutting liquid comprises 2.5% of triisopropylsilane, 2.5% of ultrapure water and 95% of trifluoroacetic acid according to volume fraction.
  9. 9. The method for preparing a polypeptide nanomaterial for down-regulating tumor dryness in combination with NRP1 according to claim 4, wherein the amidation reaction is performed at a temperature of 38 ℃ to 45 ℃ for 36h to 48h.
  10. 10. Use of a polypeptide nanomaterial for down-regulating tumor stem in combination with NRP1 according to any of claims 1-3 in the preparation of a medicament for blocking NRP1 pathway.

Description

Polypeptide nano material for reducing tumor dryness by combining NRP1 as well as preparation method and application thereof Technical Field The invention relates to the technical field of nano materials, in particular to a polypeptide nano material for reducing tumor dryness by combining NRP1, a preparation method and application thereof. Background Neuropilin-1 (NRP 1) is a key molecule that maintains tumor stem cell characteristics and promotes malignant progression of tumors. In triple negative breast cancer, VEGFA secreted by tumor-associated macrophages activates a downstream GAPVD 1/Wnt/beta-catenin signal pathway through an NRP1 receptor, so that the stem cell characteristics of tumor cells are obviously enhanced, and the expression of stem cell markers such as CD44, OCT-4, nanog, SOX-2 and the like is up-regulated, and meanwhile, the formation capacity of tumor balls is promoted. It is noted that VEGFA expressed by tumor cells themselves can also maintain their stem cell properties in an autocrine manner through NRP1 and form a positive feedback cycle with macrophages in the tumor microenvironment, which together drive the maintenance of tumor stem properties. Furthermore, studies in hepatocellular carcinoma demonstrated that Sema3C promotes tumor stem maintenance, self-renewal, and chemotherapy resistance by binding to NRP1 receptor, activating the downstream AKT/Gli1/C-Myc signaling axis. These findings indicate that NRP1 maintains tumor stem cell characteristics through multi-mechanism, multicellular type synergy, making it a potentially important target for targeting tumor stem and overcoming therapeutic resistance. Current therapeutic strategies for NRP1 remain significantly inadequate. First, early anti-NRP 1 mab (e.g., MNRP 1685A) was terminated in clinical studies due to severe adverse reactions (e.g., proteinuria). In addition, NRP1 acts as a multi-ligand, multifunctional co-receptor, the signaling pathway of which is complex, and existing antibodies have difficulty in fully blocking their interactions with multiple ligands (e.g., VEGF, sema3A, sema4A, TGF- β, etc.), potentially leading to limited therapeutic efficacy or functional redundancy. Accordingly, the prior art is still in need of improvement and development. Disclosure of Invention In view of the above-mentioned shortcomings of the prior art, the present invention aims to provide a polypeptide nanomaterial for binding NRP1 to down regulate tumor dryness, and a preparation method and application thereof, which aim to solve the problem that the existing antibody is difficult to block interaction of NRP1 as a co-receptor with various ligands. The technical scheme of the invention is as follows: A polypeptide nanomaterial for combining NRP1 to down regulate tumor dryness comprises a polypeptide with an amino acid sequence shown as SEQ ID NO.1 and a hydrophobic fluorescent group connected with last lysine of the polypeptide through an amide bond, wherein the polypeptide comprises a ligand peptide of NRP1 and a fiber deformation primitive connected with the ligand peptide through an amide bond, and the hydrophobic fluorescent group is connected with one end of the fiber deformation primitive, which is far away from the ligand peptide, through the last lysine. The polypeptide nanomaterial for reducing tumor dryness by combining with NRP1 is characterized in that the hydrophobic fluorescent group is selected from one of protoporphyrin, 7-nitrobenzodiazole and dipyrene molecules. The polypeptide nanomaterial for reducing tumor dryness by combining with NRP1 comprises the following structural formula: 。 A method for preparing a polypeptide nanomaterial for down-regulating tumor dryness in combination with NRP1, comprising the steps of: Swelling the chlorine resin to obtain a first product; Mixing K [ Fmoc-Lys (Boc) -OH ], N-diisopropylethylamine and N, N-dimethylformamide, and then reacting with the first product to obtain a second product; A mixed system of methanol, N-diisopropylethylamine and N, N-dimethylformamide is adopted to react with the second product, so as to obtain a third product; Coupling is sequentially carried out on the main chain of the third product according to the sequence of aspartic acid D, alanine A, lysine K (Boc), leucine L, cysteine C, lysine K (Boc), leucine L, valine V, phenylalanine F and lysine K (Dde) to obtain a fourth product; removing a main chain amino Fmoc protecting group in the fourth product, and using acetic anhydride to end-cap the amino exposed by the main chain to obtain a fifth product; removing a side chain amino Dde protecting group of lysine K (Dde) in the fifth product by using an N, N-dimethylformamide solution containing hydrazine hydrate to obtain a sixth product; Mixing a hydrophobic fluorescent group, an additive, N' -diisopropylcarbodiimide and N, N-dimethylformamide with the sixth product, and carrying out amidation reaction to obtain the nano polypeptide material. The preparation me