CN-120554367-B - Molecular probe design for tumor microenvironment response and photodynamic enhanced treatment and preparation method thereof

Abstract

The invention belongs to the field of organic molecular fluorescent probes, and reasonably designs a small molecular fluorescent probe HX, and diagnosis and treatment integration is realized by monitoring relevant physical properties of tumor microenvironment and photodynamic treatment of tumors. The tumor microenvironment of the solid tumor generally has the characteristics of low pH, high viscosity, low oxygen and the like, and has important significance for monitoring and effectively inhibiting tumor tissues in real time based on the characteristic parameters of tumor parts. Here we have devised a near infrared fluorescent probe HX that is not responsive to ambient viscosity changes in high pH environments, whereas in tumor low pH microenvironments, its "Donor-pi-acceptor (Donor-pi-Acceptor, D-pi-a)" structure is opened, exhibiting sensitive viscosity response characteristics, thus enabling dynamic monitoring of tumor microenvironments. In addition, through structural optimization, the probe can be used as a photosensitizer with I-type reaction for photodynamic therapy of tumor tissues under the condition of hypoxia. By means of a mouse living tumor model, the probe HX has the capability of identifying tumor tissues and normal tissues, has an excellent tumor inhibition effect and good biocompatibility under the condition of hypoxia, and provides a new technology for diagnosis and treatment integrated design of tumor diseases.

Inventors

• YU CHANGMIN
• ZHANG LONG
• WU HANBIN
• Chen Zena
• HUANG ZHONGXI
• SHEN QIAN

Assignees

• 南京工业大学

Dates

Publication Date

20260512

Application Date

20250619

Claims (5)

1. 1. The probe molecule HX with tumor microenvironment response and photodynamic therapy function is characterized in that the structural formula of the probe is shown as formula I: Formula I.
2. 2. A method for preparing a probe molecule HX according to claim 1, characterized by the steps of: adding sodium metabisulfite, dimethylamine and 2, 7-dihydroxynaphthalene into water, stirring and heating the mixture to 150 ℃ in a pressure-resistant bottle, reacting for 6 hours, cooling and adjusting the pH to 6 by using hydrochloric acid of 2M, waiting for complete precipitation of solid, filtering and drying, and purifying by using silica gel column chromatography to obtain a white crystal compound 1 ; Adding the compound 1 into N, N-dimethylformamide solution under the protection of nitrogen, adding phosphorus oxychloride after ice bath for 0.5 hour, stirring for 0.5 hour under ice bath, reacting for 4 hours at 50 ℃, adding ice water after the reaction is finished, adjusting the pH of the solution to 8 under ice bath by using sodium bicarbonate, filtering and washing with pure water after the solid is completely separated out, and drying to finally obtain the yellowish green solid compound 2 ; Adding 2,3, 3-trimethyl-4, 5-benzo-3H-indole into anhydrous acetonitrile under the protection of nitrogen, stirring and heating to 60 ℃ for reaction for 6 hours after the ethyl iodide is added, adding anhydrous diethyl ether for recrystallization after the reaction is finished, filtering and drying to obtain a grey green solid compound 3 ; And fourthly, dissolving the compound 2 and the compound 3 in absolute ethanol under the environment of nitrogen protection, refluxing for 8 hours at 80 ℃, decompressing and distilling the ethanol solvent by a rotary evaporator, and purifying by silica gel column chromatography to obtain the probe molecule HX.
3. 3. Use of a probe molecule HX according to claim 1 for the preparation of a medicament for diagnosing a change in pH and viscosity of a tumor microenvironment.
4. 4. Use of a probe molecule HX according to claim 1 for the preparation of a medicament for photodynamic therapy of tumors.
5. 5. The use of claim 3 or 4, wherein the tumor is a tumor in a mouse breast cancer model.

Description

Molecular probe design for tumor microenvironment response and photodynamic enhanced treatment and preparation method thereof Technical Field The invention belongs to the field of organic molecular fluorescent probes, and particularly discloses a molecular probe capable of being used for living tumor microenvironment response and photodynamic enhanced treatment, and a preparation method and application thereof. Background Tumor microenvironments often have low pH, high viscosity, hypoxia, etc., which are closely related to the development, metastasis and spread of malignant tumors, and monitoring pH/viscosity changes of specific organelles is critical for early detection of cancer, undoubtedly beneficial for improving therapeutic effects. Small molecule fluorescent probes have become powerful tools for dynamic and visual detection of tumor cell viscosity, most viscosity sensitive probes have a "Donor-pi-acceptor (Donor-pi-Acceptor, D-pi-a)" framework in which the molecular rotor promotes the appearance of intramolecular charge transfer (Twisted intramolecular CHARGE TRANSFER, tic) tortuosity, thereby enhancing their sensitivity to viscosity changes. However, such D-pi-a probes are susceptible to intramolecular charge transfer (Intramolecular CHARGE TRANSFER, ICT) effects, which can present challenges for false positive signals in complex cellular microenvironments, affecting the Signal-to-background ratio (SBR). To solve this problem, fluorescent probe designs based on "double-lock" strategies, in particular on tumor microenvironment-related multiparameter responses, are very effective solutions, but have been reported recently. The near infrared fluorescent probe with the pH value locked and the tumor viscosity response is prepared, the probe does not respond to the surrounding viscosity change in a high pH value environment, however, in a tumor low pH value microenvironment, the D-pi-A structure is opened, and the sensitive viscosity response characteristic is shown, so that the dynamic monitoring of the tumor microenvironment is realized. In addition, traditional photosensitizers are type II photodynamic (Photodynamic therapy, PDT) mechanisms that kill cancer cells by photoinduction of reactive oxygen species (Reactive oxygen species, ROS) at high concentrations of O 2. However, the lack of oxygen deficient microenvironment of solid tumors and the increased consumption of O 2 by O 2 during PDT severely limits the therapeutic efficacy. The development of novel type I photosensitizers can well solve the above problems. The type I photosensitizer directly reacts with a substrate (such as water and oxygen) under the illumination to generate superoxide radical (O 2•−), hydroxyl radical (. OH) and other cation or anion radicals, so that less O 2 is consumed, and the photosensitizer is suitable for a deep anoxic environment of tumor tissues. Disclosure of Invention In order to solve the problems, the invention designs the double-lock fluorescent probe molecule HX, which monitors the viscosity change in the tumor slightly acidic environment in real time, and proves that the probe molecule has an I-type PDT mechanism through molecular structure regulation and control, thereby enhancing the tumor treatment effect and realizing the integration of diagnosis and treatment. In order to achieve the above purpose, the invention discloses the following technical scheme: In a first aspect, the invention provides a probe molecule with tumor microenvironment response and photodynamic therapy function, wherein the probe molecule is denoted as HX, and the structure of the probe molecule is shown as formula I: A formula I; Under alkaline conditions, the hydroxyl group of naphthalene ring in the probe molecule HX and the spiro-benzofuran structure undergo a1, 4-cycloaddition reaction to form a neutral six-membered ring structure, and the non-fluorescence is shown due to the photoinduced electron transfer effect. However, this structure is destroyed under acidic conditions, resulting in the pyridine-like ring of the probe opening. At this stage, the probe has a D-pi-A framework, thereby enhancing its sensitivity to viscosity changes. Thus, HX has a pH switch and the ability to detect tumor microenvironment viscosity. In a second aspect, the preparation method of the probe molecule HX comprises the following steps: The first step is to add sodium metabisulfite, dimethylamine and 2, 7-dihydroxynaphthalene into water, stir and heat the mixture to 150 ℃ in a pressure-resistant bottle, react for 6 hours, cool and adjust the pH to 6 with 2M hydrochloric acid, wait for complete precipitation of solid, filter and dry, then purify by silica gel column chromatography to obtain white crystalline compound 1-7-dimethylamino-2-naphthol. And secondly, adding the compound 1-7-dimethylamino-2-naphthol into N, N-dimethylformamide solution under the protection of nitrogen, adding phosphorus oxychloride after ice bath for 0.5 hour, continuously sti