Doxorubicin derivative as well as preparation and application thereof

Abstract

The invention discloses an adriamycin derivative as well as preparation and application thereof. According to the invention, 2, 4-dinitrobenzenesulfonyl chloride with SO2 release behavior of glutathione in tumor cells is selected as a drug molecule, and the drug molecule DOX-DNs is prepared from the drug molecule and a traditional chemotherapeutic drug adriamycin through a chemical coupling method. The preparation method comprises the following steps: respectively dissolving 2, 4-dinitrobenzenesulfonyl chloride in an organic solvent to obtain a first solution and a second solution; adding organic alkali into the first solution, stirring in an ice-water bath, slowly adding the second solution to react, filtering a reaction product, and taking a solid to obtain the adriamycin derivative. According to the preparation method disclosed by the invention, the compound is further loaded into a pH response type degradable porous metal framework material ZIF-8 carrier to prepare an anti-tumor nano therapeutic agent (DOX-DNs (at) ZIF-8), and efficient treatment on drug-resistant tumor cells is realized by combining SO2 gas molecules with a chemotherapeutic drug DOX.

Description

Technical Field

[0001] The present invention relates to an anti-tumor compound, in particular to an adriamycin derivative and the preparation and application thereof. Background Art

[0002] At present, cancer treatment includes surgical treatment, drug treatment, chemotherapy and other methods. However, these traditional treatment methods are harmful to the human body, have poor treatment effects, and have high recurrence rates. Therefore, it is urgent to find an efficient and inexpensive treatment method. In recent years, the method of using nanomaterials to encapsulate drugs to form nanotherapeutic agents for the treatment of cancer has become popular. It uses a nano drug delivery system to deliver chemotherapy drugs into tumor cells, thereby solving the problem of intracellular delivery of chemotherapy drugs. For example, in nanocarriers, 2-methylimidazole and Zn 2+ The metal-organic framework nanomaterial (ZIF-8) composed of it is more widely used as a drug carrier. It can exist stably in the body, but in the acidic environment inside cancer cells, ZIF-8 can decompose and release drugs, thereby improving the treatment of cancer.

[0003] Doxorubicin is an anti-tumor drug that can inhibit the synthesis of RNA and DNA. It has the strongest inhibitory effect on RNA, a wide anti-tumor spectrum, and is effective against a variety of tumors. It is a non-specific drug for the cycle and has a killing effect on tumor cells in various growth cycles. It is mainly suitable for acute leukemia, and is effective for acute lymphocytic leukemia and granulocytic leukemia, and is generally used as a second-line drug. Although doxorubicin is an anti-tumor antibiotic widely used in clinical practice, the resistance of tumor cells to doxorubicin has become a major obstacle to the success of chemotherapy. From a mechanistic point of view, on the one hand, transport proteins such as P-gp protein on the tumor cell membrane are overexpressed, which acts like a "pump" to actively expel doxorubicin that enters the cell, so that the intracellular drug concentration cannot reach an effective killing dose. On the other hand, the antioxidant system in the cell is activated to neutralize the free radicals produced by doxorubicin and weaken its damage to tumor cells. At the same time, the DNA repair ability of tumor cells is enhanced, and even if doxorubicin causes DNA damage, it can be quickly repaired. Clinically, doxorubicin resistance leads to poor chemotherapy effects and increased risk of recurrence in patients. To overcome this problem, combination drug strategies are currently being studied, using other drugs to inhibit resistance-related proteins, or developing new doxorubicin analogs, aiming to restore the powerful effectiveness of this anti-cancer "weapon".

[0004] As an emerging tumor treatment technology, gas therapy mainly uses therapeutic gases [nitric oxide (NO), carbon monoxide (CO), hydrogen sulfide (H2S), sulfur dioxide (SO2), etc.] to replace traditional chemotherapy drugs. Sulfur dioxide (SO2) gas therapy has shown great potential in the field of tumor treatment. It can disrupt the redox balance within tumor cells, leading to an increase in the concentration of reactive oxygen species in tumor cells, causing mitochondrial dysfunction and oxidative damage to biological macromolecules, and then inducing apoptosis of tumor cells. Research has shown that gas molecules exhibit unique potential in the treatment of drug-resistant tumors. For example, gases such as nitric oxide (NO) have multiple mechanisms of action. On the one hand, it can regulate the tumor microenvironment, improve the abnormal state of blood vessels, enable drugs to be better delivered to the tumor site, and overcome the drug resistance problem caused by poor blood flow; on the other hand, NO can directly interfere with the drug-resistant related signaling pathways within tumor cells, inhibit the synthesis and function of drug-resistant proteins, and enhance the sensitivity of tumor cells to traditional chemotherapy drugs. In recent years, SO2 gas has been emerging in the treatment of drug-resistant tumors. It has been found that SO2 can precisely act on the abnormal metabolic pathways within tumor cells, interfere with their energy supply, starve the tumor cells that rely on special metabolism to maintain drug resistance, and then inhibit tumor growth. SO2 can also cooperate with some traditional chemotherapy drugs. By changing the permeability of the tumor cell membrane, it helps drugs enter the cells more efficiently to exert their efficacy, breaking through the drug resistance barrier of tumor cells, and providing an innovative strategy for the treatment of drug-resistant tumors. Summary of the Invention

[0005] Objectives of the Invention: The objective of the present invention is to provide an adriamycin derivative or a pharmaceutically acceptable salt thereof to solve the problem of adriamycin drug resistance. Another objective of the present invention is to propose a preparation method of the adriamycin derivative or a pharmaceutically acceptable salt thereof to solve the problem of how to prepare the adriamycin derivative or a pharmaceutically acceptable salt thereof. The third objective of the present invention is to propose the application of the adriamycin derivative in the preparation of anti-tumor nano-formulations to solve the problem of how to prepare anti-tumor nano-formulations. The fourth objective of the present invention is to propose the application of the adriamycin derivative in the preparation of drugs for inhibiting the expression of the drug transporter P-gp to solve the problem of how to prepare drugs for inhibiting the expression of the drug transporter P-gp.

[0006] Technical Solution: An adriamycin derivative or a pharmaceutically acceptable salt thereof described in the present invention has the following structural formula:

[0007]

[0008] The second aspect of the present invention discloses a preparation method of the above adriamycin derivative or a pharmaceutically acceptable salt thereof, including the following reactions:

[0009]

[0010] In the present invention, a drug molecule DOX-DNs based on doxorubicin derivative was synthesized by chemically coupling the chemotherapeutic drug doxorubicin (DOX) and the sulfur dioxide prodrug molecule 2,4-dinitrobenzenesulfonyl chloride (DNs), and the compound has good water solubility. To improve the efficiency of the nano-drug entering tumor cells, the porous metal-organic framework nano-carrier ZIF-8 was used to load the drug molecule to form a nano-therapeutic agent (DOX-DNs@ZIF-8). This nano-therapeutic agent can enter tumor cells through endocytosis, and is degraded in the acidic tumor microenvironment to release the drug molecule DOX-DNs, which then reacts with glutathione (GSH) in the tumor cells to release SO2 gas molecules and DOX. The released SO2 can act on mitochondria and inhibit the expression of the drug transporter P-gp, improve the sensitivity of drug-resistant tumor cells MCF-7 / MDR to the chemotherapeutic drug DOX, enhance the chemotherapeutic effect and combine with SO2 gas therapy to achieve efficient treatment of drug-resistant tumors. In addition, the anti-tumor nano-therapeutic agent of the present invention will not be degraded in normal cells, so it will not produce obvious toxic side effects.

[0011] Preferably, the specific preparation method includes the following operation steps:

[0012] (1) Dissolve doxorubicin in an organic solvent to obtain a first solution, and dissolve 2,4-dinitrobenzenesulfonyl chloride in an organic solvent to obtain a second solution;

[0013] (2) Add an organic base to the first solution and stir in an ice-water bath, then slowly add the second solution to react, and filter the reaction product to obtain a solid to get the doxorubicin derivative.

[0014] Preferably, in step (1), the molar ratio of doxorubicin to 2,4-dinitrobenzenesulfonyl chloride is 1:1 - 1.5, and the organic solvent includes at least one of tetrahydrofuran, dimethyl sulfoxide, dichloromethane, and dimethylformamide.

[0015] Preferably, in step (1), the first solution is a tetrahydrofuran solution of doxorubicin with a concentration of 100 - 120 mg / mL, and the second solution is a tetrahydrofuran solution of 2,4-dinitrobenzenesulfonyl chloride with a concentration of 45 - 55 mg / mL.

[0016] Preferably, in step (2), the organic base is triethylamine or diisopropylethylamine or DBU, and the dosage of the organic base is 1.0 - 1.2% of the mass of doxorubicin.

[0017] The third aspect of the present invention discloses the application of the above-mentioned doxorubicin derivative in the preparation of an anti-tumor nano-therapeutic agent.

[0018] The method for preparing an anti-tumor nano-therapeutic agent using the above-mentioned doxorubicin derivative includes the following steps:

[0019] Dissolve the adriamycin derivative, zinc nitrate and 2-methylimidazole in water, stir the reaction vigorously, and centrifuge the reaction product to remove the supernatant to obtain an anti-tumor nano-therapeutic agent.

[0020] Preferably, the molar ratio of the adriamycin derivative, zinc nitrate and 2-methylimidazole is 0.5-2:10:25-30, and the reaction conditions are stirring reaction at room temperature for 4-8 h.

[0021] The molar ratio of the adriamycin derivative, zinc nitrate and 2-methylimidazole is further preferably 1:10:25-30.

[0022] The anti-tumor nano-therapeutic agent synthesized in the present invention is a monodisperse spherical nano-particle with a particle size range of 120-150 nm; the loading rate of the drug molecule DOX-DNs in the synthesized anti-tumor nano-therapeutic agent is 40% by mass fraction.

[0023] The fourth aspect of the present invention discloses the application of the above adriamycin derivative in the preparation of a drug for inhibiting the expression of the drug transporter P-gp.

[0024] Beneficial effects: Compared with the prior art, the present invention has the following remarkable advantages:

[0025] The drug molecule (DOX-DNs) synthesized in the present invention has a "double effect of killing two birds with one stone": one is that under the action of GSH in tumor cells, SO2 gas molecules can be released, and the released SO2 gas can act on mitochondria to generate reactive oxygen species, promoting apoptosis of tumor cells; the other is that the SO2 gas molecules can also inhibit the expression of the drug transporter P-gp, reduce the efflux of the chemotherapeutic drug DOX, improve the sensitivity of drug-resistant tumor cells to the chemotherapeutic drug DOX, and enhance the chemotherapeutic effect. By combining SO2 gas molecules with the chemotherapeutic drug DOX, efficient treatment of drug-resistant tumor cells is achieved.

[0026] The present invention further combines DOX-DNs with the pH-responsive nano-carrier ZIF-8, and the anti-tumor nano-therapeutic agent (DOX-DNs@ZIF-8) invented can enter tumor cells in large amounts through endocytosis, and realizes efficient treatment of drug-resistant tumors by synergistically combining chemotherapy and gas therapy. Description of the Drawings

[0027] Figure 1 1H NMR spectrum of the conjugate drug DOX-DNs;

[0028] Figure 2 Scanning electron micrograph of the anti-tumor nano-therapeutic agent DOX-DNs@ZIF-8;

[0029] Figure 3Fluorescence microscope images of PBS, DOX, and DOX-DNs@ZIF-8 after incubation with MCF-7 / MDR drug-resistant cells for 30 min after adding a sulfur dioxide fluorescence probe;

[0030] Figure 4 Levels of P-gp protein expression detected by Western blot after co-culture of DOX, DOX-DNs, and DOX-DNs@ZIF-8 with MCF-7 / MDR drug-resistant tumor cells;

[0031] Figure 5 Toxicity of different experimental groups to MCF-7 / MDR drug-resistant tumor cells;

[0032] Figure 6 Therapeutic effects of the nano-therapeutic agent DOX-DNs@ZIF-8 on different types of tumor cells. Specific implementation manners

[0033] The technical solution of the present invention will be further described below with reference to the accompanying drawings.

[0034] Example 1: The structural formula of an adriamycin derivative DOX-DNs is as follows:

[0035]

[0036] The preparation method of DOX-DNs is as follows:

[0037]

[0038] First, weigh 540 mg of adriamycin and dissolve it in a round-bottom flask containing 5 mL of tetrahydrofuran solution. Then, add 5.4 mg of triethylamine and stir in an ice-water bath for 30 min. Next, slowly add 5 mL of tetrahydrofuran solution dissolved with 266 mg of 2,4-dinitrobenzenesulfonyl chloride and continue stirring for 5 h. After the reaction is completed, filter to obtain a solid, wash it with tetrahydrofuran, and the red solid obtained is the adriamycin derivative DOX-DNs; among them, the yield of the adriamycin derivative DOX-DNs is 75.2%. The nuclear magnetic resonance hydrogen spectrum of the adriamycin derivative DOX-DNs (room temperature, 400 MHz, DMSO-d6) is as Figure 1 shown.

[0039] 11H NMR (400 MHz, DMSO) δ 13.98 (s, 1H), 13.18 (s, 1H), 8.28–7.52 (m, 6H), 5.50 (s, 2H), 5.28 (s, 1H), 4.91 (dd, J = 15.3, 9.5 Hz, 2H), 4.60 (d, J = 5.4 Hz, 2H), 4.20 (d, J = 6.5 Hz, 1H), 3.97 (s, 3H), 3.61 (d, J = 3.5 Hz, 1H), 2.97 (d, J = 18.2 Hz, 1H), 2.79 (d, J = 18.2 Hz, 1H), 2.16 (d, J = 14.2 Hz, 1H), 2.04 (dd, J = 29.5, 9.4 Hz, 1H), 1.88 (t, J = 11.1 Hz, 1H), 1.68 (d, J = 8.8 Hz, 1H), 1.11 (dd, J = 36.3, 5.9 Hz, 3H).

[0040] The method for preparing an anti-tumor nanoformulation using DOX-DNs is as follows:

[0041] Dissolve 90 mg of the doxorubicin derivative DOX-DNs prepared by the above method in 20 mL of water, then add the solution to a 50 mL round-bottom flask, and further add 189 mg of zinc nitrate and 205.3 mg of 2-methylimidazole. After vigorously stirring for 4 h, centrifuge to remove the supernatant to obtain the nano-therapeutic agent (DOX-DNs@ZIF-8), wherein the drug loading rate is about 48% by mass fraction.

[0042] Example 2: The preparation methods of the doxorubicin derivative and the anti-tumor nanoformulation are as follows:

[0043] (1) First, weigh 500 mg of doxorubicin and dissolve it in a 50 mL round-bottom flask containing 5 mL of tetrahydrofuran solution. Then add 6.0 mg of triethylamine and stir in an ice-water bath for 30 min. Then slowly add a 5 mL tetrahydrofuran solution containing 367 mg of 2,4-dinitrobenzenesulfonyl chloride and continue stirring for 5 h. After the reaction is completed, filter to obtain a solid, wash it with tetrahydrofuran, and the red solid obtained is the doxorubicin derivative DOX-DNs; wherein, the yield of the doxorubicin derivative DOX-DNs is 79.5%;

[0044] (2) Dissolve 90 mg of the doxorubicin derivative DOX-DNs obtained in step (1) in 20 mL of water, then add the solution to a 50 mL round-bottom flask, and further add 74 mg of zinc nitrate and 205.3 mg of 2-methylimidazole. After vigorously stirring for 4 h, centrifuge to remove the supernatant to obtain the nano-therapeutic agent (DOX-DNs@ZIF-8), wherein the drug loading rate is about 45% by mass fraction.

[0045] Example 3: The preparation method of adriamycin derivative and anti-tumor nano preparation is as follows:

[0046] (1) First, weigh 600 mg of adriamycin, dissolve it in 5 mL of tetrahydrofuran solution, add it to a 50 mL round-bottom flask, then add 7.2 mg of triethylamine, and stir in an ice-water bath for 30 min. Then slowly add 5 mL of tetrahydrofuran solution dissolved with 440 mg of 2,4-dinitrobenzenesulfonyl chloride, and continue to stir for 5 h. After the reaction is completed, filter to obtain a solid, wash it with tetrahydrofuran, and a red solid, namely adriamycin derivative DOX-DNs, can be obtained; among them, the yield of adriamycin derivative DOX-DNs is 82%;

[0047] (2) Dissolve 90 mg of the adriamycin derivative DOX-DNs obtained in step (1) with 20 mL of water, then add the solution to a 50 mL round-bottom flask, add 74 mg of zinc nitrate and 205.3 mg of 2-methylimidazole, stir vigorously for 4 h, and centrifuge to remove the supernatant to obtain a nano-therapeutic agent (DOX-DNs@ZIF-8), and the drug loading rate is about 43% by mass fraction.

[0048] Characterize the morphology, mechanism of action, intracellular sulfur dioxide release and anti-tumor effect of the anti-tumor therapeutic agent DOX-DNs@ZIF-8 obtained in Example 1.

[0049] 1. Morphology characterization of nano-therapeutic agent DOX-DNs@ZIF-8

[0050] Characterize the morphology of the synthesized nano-therapeutic agent DOX-DNs@ZIF-8, as Figure 1 shown by the scanning electron microscope results. The finally prepared nano-therapeutic agent is a monodisperse spherical nano-particle with a relatively rough surface, and its particle size range is 120-150 nm.

[0051] 2. Fluorescence imaging of intracellular sulfur dioxide gas

[0052] Seed drug-resistant breast cancer cells (MCF-7 / MDR) in a 6-well plate (5×10 4 cells / mL -1 , 2 mL per well), and culture them in a 37 °C, 5% CO2 incubator for 24 h. Add PBS, DOX (final concentration 100 μg / mL), and the nano-therapeutic agent DOX-DNs@ZIF-8 (100 μg / mL) obtained in Example 1 respectively. After incubating the cells in the corresponding solution for 4 h, add a sulfur dioxide molecular fluorescence probe and incubate for 30 min. Then remove the culture medium and wash the cells 3 times with phosphate buffer. Finally, observe the cells by confocal laser scanning microscope.

[0053] The results are as Figure 2As shown, no blue fluorescence was found in the cells cultured with PBS and DOX, indicating that sulfur dioxide gas was not produced in the MCF-7 / MDR cells cultured under these conditions. Obvious blue fluorescence was found in the cells co-cultured with the nano-therapeutic agent DOX-DNs@ZIF-8, indicating that DOX-DNs@ZIF-8 entered the tumor cells and could release sulfur dioxide gas under the action of GSH in their microenvironment.

[0054] 3. Sulfur Dioxide Gas Molecule Inhibits the Expression of Drug Transporter P-gp

[0055] The drug-resistant breast cancer cells (MCF-7 / MDR) were incubated in Dulbecco's Modified Eagle Medium (DMEM). This medium contained 10% fetal bovine serum and 1% penicillin-streptomycin. The MCF-7 / MDR cells were seeded into 24-well plates (5×10 4 cells / mL -1 , 0.1 mL per well), and cultured in an incubator at 37 °C and 5% CO2 for 24 h. Then, DOX, DOX-DNs, and the prepared nano-therapeutic agent DOX-DNs@ZIF-8 with a final concentration of 100 μg / mL were added respectively and cultured for 24 h. The expression level of P-gp protein in MCF-7 / MDR cells treated differently was determined by Western blot.

[0056] The results are as Figure 4 shown. The Western blot results showed that compared with the DOX alone group, the expression of P-gp protein in the tumor cells treated with DOX-DNs was inhibited to a certain extent. This was because the drug DOX-DNs could release SO2 gas in the tumor cells, thus inhibiting the expression of P-gp protein in the cells. In contrast, the expression of P-gp protein in the cells treated with the nano-therapeutic agent DOX-DNs@ZIF-8 was greatly inhibited. This was because the nano-carrier increased the efficiency of the nano-drug DOX-DNs entering the tumor cells, and then the concentration of SO2 gas released in the tumor cells increased, resulting in the maximum inhibition of the expression of P-gp protein.

[0057] 4. Cytotoxicity Characterization

[0058] The drug-resistant breast cancer cells (MCF-7 / MDR) were incubated in Dulbecco's Modified Eagle Medium (DMEM). This medium contained 10% fetal bovine serum and 1% penicillin-streptomycin. The MCF-7 / MDR cells were seeded into 96-well plates (5×10 4 cells / mL -1, in each well (0.1 mL), and cultured in an incubator at 37 °C and 5% CO2 for 24 h. Then, ZIF-8, DOX, and the prepared nano-therapeutic agent DOX-DNs@ZIF-8 were added respectively and co-cultured for 24 h. The survival rate of the corresponding cells was measured by the MTT method. The drug concentration range was 0 - 160 μg / mL.

[0059] The results are as Figure 5 shown. It can be seen from the results in the figure that ZIF-8 alone did not produce obvious toxicity to tumor cells. Even at a concentration as high as 160 μg / mL, the cell survival rate remained above 97%, indicating that the nano-carrier ZIF-8 we used has excellent biocompatibility and will not produce obvious toxicity to tumor cells as a nano-drug carrier itself. The single chemotherapy drug DOX has certain toxicity to drug-resistant MCF-7 / MDR cells. However, the effect is not very obvious. Even at a concentration of 160 μg / mL, the survival rate of MCF-7 / MDR cells still remained at about 70%. This is because after the chemotherapy drug DOX enters drug-resistant MCF-7 / MDR cells, it can be transported by the P-gp drug transporter expressed in the cells to the extracellular, reducing the intracellular drug concentration and thus weakening the chemotherapy effect. Even if the concentration of the chemotherapy drug is increased, the treatment effect is still not very obvious. However, the synthesized nano-therapeutic agent DOX-DNs@ZIF-8 showed excellent therapeutic effects on drug-resistant MCF-7 / MDR cells. At a concentration of 160 μg / mL, the survival rate of MCF-7 / MDR cells was reduced to about 5%. This shows that the nano-therapeutic agent can improve the efficiency of the drug DOX-DNs entering tumor cells, and combine with the SO2 gas and chemotherapy drug DOX released in tumor cells, and by inhibiting the expression of P-gp protein, improve cell drug resistance, and thus achieve the therapeutic effect on drug-resistant MCF-7 / MDR cells. Compared with the control group, the nano-therapeutic agent DOX-DNs@ZIF-8 reported in the present invention can greatly improve the chemotherapy effect of drug-resistant tumor cells.

[0060] 5. Anti-tumor universality test

[0061] Human cervical cancer cells (HeLa), mouse breast cancer cells (4T1), rat glioma cells (C6), and human normal liver cells (7702) were incubated in Dulbecco's Modified Eagle Medium (DMEM). The medium contained 10% fetal bovine serum and 1% penicillin-streptomycin. HeLa cells, 4T1 cells, C6 cells, and 7702 cells were seeded into 96-well plates (5×10 4 cells / mL -1, in each well (0.1 mL), and cultured in an incubator at 37 °C with 5% CO2 for 24 h. Then the cells were grown in the nano-therapeutic agent for 24 h. The survival rate of the corresponding cells was determined by the MTT method.

[0062] The results are as Figure 6 shown. The nano-therapeutic agent DOX-DNs@ZIF-8 did not produce obvious cytotoxicity to human normal liver cells (7702). Even at a concentration as high as 160 μg / mL, the 7702 cells still maintained a survival rate of 95%. This is because the interior of human normal liver cells is weakly alkaline, and the nano-therapeutic agent DOX-DNs@ZIF-8 can stably exist in a weakly alkaline environment, and the loaded drug molecule DOX-DNs will not be released, thus having less cytotoxicity to cells. However, in human cervical cancer cells (HeLa), mouse breast cancer cells (4T1), and rat glioma cells (C6), the nano-therapeutic agent DOX-DNs@ZIF-8 at different concentrations showed different degrees of cytotoxicity to the cells. At a concentration of 160 μg / mL, the mortality rate of the cells could reach over 80%. This is because in the weakly acidic environment of cancer cells, the nano-therapeutic agent DOX-DNs@ZIF-8 can be degraded and release DOX-DNs, which then react with the overexpressed GSH in tumor cells to release SO2 gas molecules. The released SO2 can act on mitochondria and inhibit the expression of the drug transporter P-gp, enhancing the chemotherapy effect and combining with SO2 gas therapy to achieve efficient treatment of tumors. The nano-therapeutic agent DOX-DNs@ZIF-8 reported in the present invention shows excellent therapeutic effects on a variety of tumor cells, indicating that the nano-therapeutic agent has certain universality for tumor treatment.

[0063] Comparative Example 1: The rest were the same as in Example 1, except that:

[0064] 2,4-Dinitrobenzenesulfonyl chloride in Example 1 was replaced with 4-nitrobenzenesulfonyl chloride, and the obtained product was as follows:

[0065]

[0066] The results of the 1H nuclear magnetic resonance spectrum of the product (room temperature, 400 MHz, DMSO-d6) were:

[0067] 11H NMR (400 MHz, DMSO) δ 8.40 (d, 2H), 8.06 (d, 2H), 7.86 - 7.82 (m, 2H), 7.36 (d, 1H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.55 (t, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0068] Comparative Example 2: The rest are the same as in Example 1, except that:

[0069] 2,4-Dinitrobenzenesulfonyl chloride in Example 1 was replaced with 2-nitrobenzenesulfonyl chloride, and the resulting product was as follows:

[0070]

[0071] The results of the 1H nuclear magnetic resonance spectrum of the product (at room temperature, 400 MHz, DMSO-d6) were as follows:

[0072] 1 1H NMR (400 MHz, DMSO) δ 8.42 (d, 1H), 8.06 - 7.97 (m, 3H), 7.86 - 7.82 (m, 2H), 7.36 (d, 1H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.55 (t, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0073] Comparative Example 3: The rest are the same as in Example 1, except that:

[0074] 2,4-Dinitrobenzenesulfonyl chloride in Example 1 was replaced with 3,4-dinitrobenzenesulfonyl chloride, and the resulting product was as follows:

[0075]

[0076] The results of the 1H nuclear magnetic resonance spectrum of the product (at room temperature, 400 MHz, DMSO-d6) were as follows:

[0077] 11H NMR (400 MHz, DMSO) δ 8.99 (s, 1H), 8.68 (d, 1H), 8.44 (d, 1H), 7.86 - 7.82 (m, 2H), 7.36 (d, 1H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.55 (t, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0078] Comparative Example 4: The rest was the same as in Example 1, except that:

[0079] 2,4 - Dinitrobenzenesulfonyl chloride in Example 1 was replaced with p - Toluenesulfonyl chloride, and the resulting product was as follows:

[0080]

[0081] The results of the 1H nuclear magnetic resonance spectrum of the product (at room temperature, 400 MHz, DMSO - d6) were as follows:

[0082] 1 1H NMR (400 MHz, DMSO) δ 7.86 - 7.82 (m, 2H), 7.66 (d, 2H), 7.39 - 7.36 (m, 3H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.55 (t, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.43 (s, 3H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0083] Comparative Example 5: The rest was the same as in Example 1, except that:

[0084] 2,4 - Dinitrobenzenesulfonyl chloride in Example 1 was replaced with 2 - Methyl - 4 - nitrobenzenesulfonyl chloride, and the resulting product was as follows:

[0085]

[0086] The results of the 1H nuclear magnetic resonance spectrum of the product (at room temperature, 400 MHz, DMSO - d6) were as follows:

[0087] 11H NMR (400 MHz, DMSO) δ 8.35 (s, 1H), 8.28 (d, 1H), 8.01 (d, 1H), 7.86 - 7.82 (m, 2H), 7.36 (d, 1H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.55 (t, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.64 (s, 3H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0088] Comparative Example 6: The rest was the same as in Example 1, except that:

[0089] 2,4 - Dinitrobenzenesulfonyl chloride in Example 1 was replaced with 2,4 - dimethylbenzenesulfonyl chloride, and the resulting product was as follows:

[0090]

[0091] The 1H - NMR spectrum of the product (at room temperature, 400 MHz, DMSO - d6) was as follows:

[0092] 1 1H NMR (400 MHz, DMSO) δ 7.86 - 7.82 (m, 2H), 7.70 (d, 1H), 7.36 - 7.32 (m, 2H), 7.24 (d, 2H), 4.69 - 4.58 (m, 4H), 3.90 (s, 3H), 3.70 (m, 1H), 3.12 (dd, 2H), 2.73 (q, 1H), 2.64 (s, 3H), 2.31 (s, 3H), 2.25 - 1.82 (m, 4H), 1.11 (d, 3H).

[0093] The adriamycin derivative samples prepared in Examples 1 - 3 and Comparative Examples 1 - 6 were respectively formulated into aqueous solutions with a concentration of 65 μg / mL. After adding the same amount of a 5 mM GSH solution and incubating for 1.5 h, 5 μM of the DEACA fluorescent probe was added. The solutions were detected using a fluorescence spectrophotometer after 8 h to measure the SO2 release amount of different compound samples. At the same time, the tumor cell inhibition rate of the anti - tumor nano - preparations prepared from different compounds at a final concentration of 160 μg / mL was detected, and the results were as follows:

[0094] Table 1 SO2 release amount and MCF - 7 / MDR cell inhibition rate of different adriamycin derivatives

[0095] Group <![CDATA[SO2 release amount (μM)]]> Inhibitory rate of MCF-7 / MDR cells (%) Example 1 51.3±2.4 93.1±2.9 Example 2 48.4±3.1 90.6±2.2 Example 3 50.6±1.8 92.4±1.5 Comparative Example 1 11.3±1.1 35.2±3.8 Comparative Example 2 14.2±1.4 37.8±3.7 Comparative Example 3 10.1±1.6 32.7±2.4 Comparative Example 4 7.8±0.9 28.9±4.3 Comparative Example 5 5.4±1.2 25.6±3.1 Comparative Example 6 13.8±1.7 38.4±2.6 Doxorubicin 0 30.7±5.6

[0096] Table 1 results show that when the number and substitution position of nitro groups are changed or nitro groups are replaced by other groups, both the release amount of SO2 and the killing effect on tumor cells are significantly reduced, indicating that the doxorubicin derivative molecule in the present invention reacts efficiently with GSH to generate SO2 depending on the double-substituted nitro groups at appropriate sites on the benzene ring, and the level of SO2 release directly affects the killing effect of doxorubicin derivatives on tumor cells.

Claims

1. An adriamycin derivative or a pharmaceutically acceptable salt thereof, characterized in that, The structural formula is as follows:

2. The preparation method of the adriamycin derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that, It includes the following reactions:

3. The preparation method of the adriamycin derivative or its pharmaceutically acceptable salt according to claim 2, characterized in that, It includes the following operation steps: (1) Dissolve doxorubicin in an organic solvent to obtain a first solution, and dissolve 2,4-dinitrobenzenesulfonyl chloride in an organic solvent to obtain a second solution; (2) Add an organic base to the first solution and stir in an ice-water bath, then slowly add the second solution for reaction, and filter the reaction product to obtain a solid to get the doxorubicin derivative.

4. The preparation method of the adriamycin derivative or its pharmaceutically acceptable salt according to claim 3, characterized in that, In step (1), the molar ratio of doxorubicin to 2,4-dinitrobenzenesulfonyl chloride is 1:1 - 1.5, and the organic solvent includes at least one of tetrahydrofuran, dimethyl sulfoxide, dichloromethane, and dimethylformamide.

5. The preparation method of the adriamycin derivative or its pharmaceutically acceptable salt according to claim 3, characterized in that, In step (1), the first solution is a tetrahydrofuran solution of doxorubicin with a concentration of 100 - 120 mg / mL, and the second solution is a tetrahydrofuran solution of 2,4-dinitrobenzenesulfonyl chloride with a concentration of 45 - 55 mg / mL.

6. The preparation method of the adriamycin derivative or its pharmaceutically acceptable salt according to claim 3, characterized in that, In step (2), the organic base is triethylamine or diisopropylethylamine or DBU, and the dosage of the organic base is 1.0 - 1.2% of the mass of doxorubicin.

7. Use of the doxorubicin derivative according to claim 1 in the preparation of an anti-tumor nano-therapeutic agent.

8. The application according to claim 7, characterized in that, It includes the following steps: Dissolve the doxorubicin derivative, zinc nitrate, and 2-methylimidazole in water, stir vigorously for reaction, and centrifuge the reaction product to remove the supernatant to obtain an anti-tumor nano-therapeutic agent.

9. The application according to claim 8, wherein The molar ratio of the doxorubicin derivative, zinc nitrate, and 2-methylimidazole is 0.5 - 2:10:25 - 30, and the reaction condition is stirring at room temperature for 4 - 8 h.

10. Use of the doxorubicin derivative according to claim 1 in the preparation of a drug for inhibiting the expression of drug transporter P-gp.

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