CN-122010919-A - Tumor targeted delivery system construction for in-situ synthesis of apoptosis inducer and application of tumor targeted delivery system construction in apoptosis-copper death cooperative treatment

Abstract

The invention belongs to the field of biological medicine, and in particular relates to a tumor targeted delivery system construction for in-situ synthesis of an apoptosis inducer and application thereof in apoptosis-copper death cooperative treatment. The invention realizes synergistic mitochondrial damage through the combined treatment of apoptosis-copper death mediated by small molecule CTT for inducing tumor cell apoptosis. CTT was synthesized in situ within tumors by bio-orthogonalization. In order to improve the selective synthesis efficiency of tumors, reduce off-target effect and overcome the problems of rapid drug clearance and metabolism, the corresponding prodrug is delivered by adopting a nano material based on ZIF-8. Under the acidic condition of tumor microenvironment, PVP@CuP@ZIF is decomposed and releases Cu (I) ions and a prodrug, and copper is catalyzed by Cu (I) to catalyze the azide-alkyne cycloaddition reaction, so that the in-situ synthesis of CTT is realized. The invention provides a novel method for overcoming tumor drug resistance by combining apoptosis and copper death and Lu Jie-mediated synergic mitochondrial injury based on in-situ synthesis with high tumor selectivity.

Inventors

• NA NA
• LI XIANG
• OUYANG JIN

Assignees

• 北京师范大学

Dates

Publication Date

20260512

Application Date

20260211

Claims (10)

1. 1. The small molecule CTT for inducing apoptosis of tumor cells is characterized by having a structural formula as follows: 。
2. 2. a method for preparing the small molecule CTT for inducing apoptosis of tumor cells according to claim 1, which comprises the steps of stirring 3-azido-7-hydroxycoumarin and 2-ethynyl thiophene in acetonitrile, carrying out reflux reaction, cooling the solution to room temperature, concentrating under reduced pressure, and purifying the crude product to obtain CTT.
3. 3. The preparation method according to claim 2, wherein the molar ratio of the 3-azido-7-hydroxycoumarin to the 2-ethynyl thiophene is 2.46:1.14, the reflux reaction is heating reaction for 24 hours at the temperature of 82 ℃, and the purification is column chromatography purification by using petroleum ether/ethyl acetate as eluent.
4. 4. A tumor selective delivery nanomaterial PVP@CuP@ZIF for synthesizing the small molecule CTT capable of inducing apoptosis of tumor cells according to claim 1 in situ is characterized in that Cu 2 O and 3-azido-7-hydroxycoumarin are encapsulated into ZIF-8 nanoparticles to form an intermediate Cu@PI@ZIF, then 2-ethynyl thiophene is anchored on the surface of the Cu@PI@ZIF to obtain the intermediate CuP@ZIF, and finally PVP is coated on the outermost layer of the CuP@ZIF to obtain the delivery nanomaterial PVP@CuP@ZIF.
5. 5. A method of preparing a tumor-selective delivery nanomaterial pvp@cup@zif as claimed in claim 4 comprising the steps of: (1) Adding sodium hydroxide into the mixture, stirring for a period of time, washing Cu 2 O nano particles, centrifuging and collecting precipitate to obtain Cu 2 O nano particles; (2) Dispersing Cu 2 O nano particles in methanol, performing ultrasonic treatment, sequentially adding a methanol solution containing 3-azido-7-hydroxycoumarin and a methanol solution containing Zn (NO 3 ) 2 ·6H 2 O) under the protection of argon, slowly adding a methanol solution containing 2-methylimidazole after uniformly stirring, stirring the mixture for reaction, centrifugally collecting precipitate, washing and drying to obtain Cu@PI@ZIF; (3) The synthesis method of the PVP@CuP@ZIF nanoparticles comprises the steps of dispersing Cu@PI@ZIF into a mixed solution of methanol and water by ultrasonic, slowly adding 2-ethynyl thiophene dissolved in the methanol into the Cu@PI@ZIF solution, stirring for a period of time, centrifugally collecting solids, washing, drying to obtain CuP@ZIF nanoparticles, adding ethanol into the CuP@ZIF ethanol solution to obtain the CuP@ZIF ethanol solution, adding the CuP@ZIF ethanol solution into the PVP solution, stirring, and centrifugally washing to obtain the PVP@CuP@ZIF nanoparticles.
6. 6. The method according to claim 5, wherein in the step (1), the molar ratio of CuI to NaAsc is 1:5, the mass ratio of CuI to sodium hydroxide is 0.025:1, and the stirring is performed at room temperature until the solution turns orange.
7. 7. The preparation method according to claim 5 or 6, wherein in the step (2), the mass ratio of the Cu 2 O nanoparticle to the 3-azido-7-hydroxycoumarin is 1:2, the concentration of the methanol solution containing the 3-azido-7-hydroxycoumarin is 2.5mg/mL, the concentration of the methanol solution containing Zn (NO 3 ) 2 ·6H 2 O) is 25mg/mL, the concentration of the methanol solution containing 2-methylimidazole is 30mg/mL, the molar ratio of the 3-azido-7-hydroxycoumarin, zn (NO 3 ) 2 ·6H 2 O) and 2-methylimidazole is 0.2-0.3:1.6-1.7:7.0-7.5, and the reaction is stirred for 0.5-2 hours at room temperature under the protection of argon.
8. 8. The preparation method of the copper-zinc alloy coated with the copper-zinc alloy is characterized in that in the step (3), the concentration of the Cu@PI@ZIF in a mixed solvent is 2.5mg/mL, the mixed solvent is composed of methanol and water in a mass ratio of 8-9:1-2 (v/v), the mass ratio of the Cu@PI@ZIF to the 2-ethynyl thiophene is 1:5-8, the concentration of the 2-ethynyl thiophene in methanol is 30mg/mL, the mass ratio of the CuP@ZIF nanoparticles to polyvinylpyrrolidone is 1:40, the concentration of the CuP@ZIF ethanol solution is 50mg/mL, after the 2-ethynyl thiophene solution is added, stirring is carried out for 4-5h at room temperature, and the CuP@ZIF nanoparticles and the PVP ZIF solution are stirred for coating at room temperature for 2-4 h.
9. 9. Use of the pvp@cup@zif of claim 1 in the preparation of a medicament for achieving tumor selective delivery for in situ synthesis of CTT, antitumor at a tumor site.
10. 10. The use according to claim 3, wherein the pvp@cup@zif is used as a delivery precursor for the synthesis of small molecule CTT inducing apoptosis of tumor cells at the tumor site via CuAAC reaction.

Description

Tumor targeted delivery system construction for in-situ synthesis of apoptosis inducer and application of tumor targeted delivery system construction in apoptosis-copper death cooperative treatment Technical Field The invention belongs to the field of biological medicine, and in particular relates to a tumor targeted delivery system construction for in-situ synthesis of an apoptosis inducer and application thereof in apoptosis-copper death cooperative treatment. Background Chemotherapy is one of the most traditional methods for treating cancer, and its mechanism of action often involves the induction of tumor cell apoptosis by small molecule drugs that regulate the mitochondrial apoptosis pathway. However, the development of novel anticancer drugs based on this approach still faces the challenges of acquired mutation and sustained drug resistance. Copper death, a non-apoptotic means of cell death, has recently been revealed to regulate cell death by mitochondrial metabolism-related mechanisms, with lower sensitivity to traditional drug resistance. Copper, when accumulated in the online granules, promotes oligomerization of the lipoylated protein and destroys the stability of the iron sulfur cluster protein, ultimately leading to protein toxic stress and cell death. Thus, combining mitochondrial apoptosis with mitochondrial copper death is expected to cause synergistic and irreversible damage to the mitochondrial region, potentially overcoming therapeutic resistance. However, this faces challenges in the delivery of small molecule compounds that induce apoptosis in tumor cells, as well as in the synergistic manner of apoptosis-copper death therapies. In addition, the efficacy of any treatment method is limited by the drawbacks of traditional drug delivery, including rapid clearance, poor tumor specificity, non-ideal pharmacokinetics, and off-target toxicity to normal tissues. Disclosure of Invention Aiming at the problems in the prior art, the invention firstly provides a small molecule CTT for inducing apoptosis of tumor cells. The invention also provides a tumor targeting delivery system for in-situ synthesis of the apoptosis inducer, namely tumor selective delivery of the nanomaterial PVP@CuP@ZIF. The invention also provides a construction method of the tumor selective delivery nanomaterial PVP@CuP@ZIF. The invention also aims to provide application of the tumor-selective delivery nanomaterial PVP@CuP@ZIF. The technical scheme adopted by the invention for achieving the purpose is as follows: The invention provides a small molecule CTT for inducing apoptosis of tumor cells, which has the structural formula: 。 The invention also provides a preparation method of the micromolecule CTT for inducing tumor cell apoptosis, which comprises the following steps of stirring 3-azido-7-hydroxycoumarin and 2-ethynyl thiophene in acetonitrile, carrying out reflux reaction, cooling the solution to room temperature, concentrating under reduced pressure, and purifying the crude product to obtain the CTT. Preferably, the molar ratio of the 3-azido-7-hydroxycoumarin to the 2-ethynyl thiophene is 2.46:1.14, the reflux reaction is heating reaction for 24 hours at the temperature of 82 ℃, and the purification is column chromatography purification by using petroleum ether/ethyl acetate as an eluent. The invention further provides a tumor selective delivery nanomaterial PVP@CuP@ZIF for synthesizing small molecule CTT capable of inducing apoptosis of tumor cells in situ, wherein the PVP@CuP@ZIF is prepared by encapsulating Cu 2 O and 3-azido-7-hydroxycoumarin into ZIF-8 nanoparticles to form an intermediate Cu@PI@ZIF, then anchoring 2-ethynyl thiophene on the surface of the Cu@PI@ZIF to obtain the intermediate CuP@ZIF, and finally coating PVP on the outermost layer of the CuP@ZIF to obtain the delivery nanomaterial PVP@CuP@ZIF. The invention also provides a preparation method of the nano material PVP@CuP@ZIF capable of being bio-orthogonalized in the tumor by selectively delivering the tumor, which comprises the following steps: (1) Adding sodium hydroxide into the mixture, stirring for a period of time, washing Cu 2 O nano particles, centrifuging and collecting precipitate to obtain Cu 2 O nano particles; (2) Dispersing Cu 2 O nano particles in methanol, performing ultrasonic treatment, sequentially adding a methanol solution containing 3-azido-7-hydroxycoumarin and a methanol solution containing Zn (NO 3)2·6H2 O) under the protection of argon, slowly adding a methanol solution containing 2-methylimidazole after uniformly stirring, stirring the mixture for reaction, centrifugally collecting precipitate, washing and drying to obtain Cu@PI@ZIF; (3) The synthesis method of the PVP@CuP@ZIF nanoparticles comprises the steps of dispersing Cu@PI@ZIF into a mixed solution of methanol and water by ultrasonic, slowly adding 2-ethynyl thiophene dissolved in the methanol into the Cu@PI@ZIF solution, stirring for a period of time, centrifugall