Carbon monoxide and iron chelator targeted combined controlled drug delivery nano-composite drug system and its preparation method

Abstract

The present invention provides a nano-composite drug system for targeted and controllable co-delivery of carbon monoxide and an iron chelator. Specifically, the structure is as follows: iron chelator - nanoparticle - exogenous metal carbonyl (ML-CO) donor - targeting group; wherein, the exogenous metal carbonyl (ML-CO) donor is a transition metal carbonyl (ML-CO) donor; and the targeting group is a triphenylphosphine derivative. This drug system has fluorescence tracing, can selectively target cancer cell mitochondria organelles, can deliver carbon monoxide under near-infrared light control, and simultaneously has the function of consuming iron ions in cancer cells. The combined administration of carbon monoxide and the iron chelator achieves a highly efficient anti-cancer effect.

Description

Technical Field

[0001] The present invention relates to the fields of biology and inorganic nano-drug. Specifically, the present invention provides a nano-composite drug system for targeted combined controlled delivery of carbon monoxide and iron chelator and its preparation method. Background Art

[0002] Iron is crucial for cell viability. It is present in proteins that perform a variety of functions, including biomolecule synthesis, oxygen transport and homeostasis, and respiration. Iron is an important component of many proteins involved in nucleic acid metabolism and repair, as well as cell cycle progression. Since iron is an essential component in anatomy and physiology and its bioavailability is very low, the iron storage in the human body is strictly controlled to ensure storage and reduce toxicity. The rapid proliferation of cancer cells leads to their need for more iron, resulting in dysregulation of the levels of key proteins in iron metabolism. Therefore, iron has become a key target for the progression and treatment of diseases including cancer.

[0003] Iron chelators have a high affinity for binding to iron. Although different types of chelators vary in structure and function, they generally contain oxygen, nitrogen, or sulfur donor atoms that form coordination bonds with iron. Deferasirox (DFX) is a synthetic tridentate iron chelator that has been approved for the treatment of thalassemia and sickle cell disease. In addition, the anti-proliferative effect of deferasirox has been demonstrated in some cancers, including hepatocellular carcinoma and myeloid leukemia, and tumor growth inhibition has been achieved in xenograft animal models of esophageal cancer and lung cancer. However, due to the small molecular weight of deferasirox, it results in a short circulation time in vivo and may also be accompanied by some serious side effects. In addition, the lack of tumor cell specificity and poor pharmacokinetics also limit its therapeutic potential and further clinical application. Therefore, conjugating deferasirox to nanoparticles is expected to improve its targeting and biocompatibility.

[0004] Currently, CO has shown great potential in the treatment of many diseases including cancer. The main target of CO is the subcellular organelle mitochondria of cancer cells, and then a series of effects are caused, such as accelerating mitochondrial respiration leading to O2 depletion, overproduction of reactive oxygen species (ROS) resulting in oxidative stress, ultimately inducing mitochondrial damage, hindering the generation of adenosine triphosphate (ATP), and promoting cancer cell apoptosis. At the same time, CO can freely cross a variety of biological membranes and tumor stroma. Therefore, when CO is combined with other anti-cancer therapies, it can serve as a good adjuvant reagent to synergistically improve the anti-cancer effect. However, how to safely deliver it to the diseased tissue remains a great challenge in the in vivo application of CO. In recent years, due to the deep tissue penetration and low phototoxicity of near-infrared light, the use of near-infrared light as an exogenous stimulus for CO release has attracted extensive attention from scientists.

[0005] Based on the above analysis, there is an urgent need for a drug system that can safely deliver CO at an appropriate concentration to the lesion tissue cells or sub-organelles, combine CO with an iron chelator, and exert an anti-proliferative effect through controllable combination drug delivery. Summary of the Invention

[0006] The object of the present invention is to provide a multifunctional nanocomposite drug system with anti-cancer effects and its preparation method.

[0007] In the first aspect of the present invention, a nanocomposite drug system is provided, and the structure of the drug system is as follows:

[0008] X - Y - Z - W

[0009] Wherein,

[0010] X represents an iron chelator;

[0011] Y represents a carrier, and the carrier is a surface - aminated nanoparticle;

[0012] Z represents an exogenous metal carbonyl (ML - CO) donor;

[0013] W represents a targeting group.

[0014] In another preferred example, the particle size of the nanocomposite drug system is 1 - 100 nm; preferably 3 - 50 nm; more preferably 3 - 8 nm.

[0015] In another preferred example, the iron chelator is selected from the group consisting of: 4 - [3,5 - bis(2 - hydroxyphenyl)-1,2,4 - triazol - 1 - yl] benzoic acid (deferasirox DFX), gallic acid, 3,4 - dihydroxybenzoic acid, deferoxamine (DFO), or a combination thereof.

[0016] In another preferred example, the nanoparticle is a heteroatom - doped graphene quantum dot; wherein the heteroatom - doped graphene quantum dot is a non - metal heteroatom - doped graphene quantum dot, and / or a graphene quantum dot co - doped with metal atoms and non - metal heteroatoms.

[0017] In another preferred example, the surface amination of the nanoparticle is obtained by reacting ethylenediamine with the nanoparticle under the activation of EDC / NHS to obtain the surface - aminated nanoparticle.

[0018] In another preferred example, the non - metal heteroatom - doped graphene quantum dots are selected from the group consisting of: nitrogen - doped graphene quantum dots, phosphorus - doped graphene quantum dots, sulfur - doped graphene quantum dots, or a combination thereof.

[0019] In another preferred example, the metal and non-metal heteroatom co-doped graphene quantum dots are selected from the group consisting of: N,P-co-doped graphene quantum dots, N,S-co-doped graphene quantum dots, Fe,N-co-doped graphene quantum dots, Mn-N-co-doped graphene quantum dots, Cu,N-co-doped graphene quantum dots, Co,N-co-doped graphene quantum dots, or combinations thereof.

[0020] In another preferred example, the nanoparticles are surface-aminated nitrogen-doped graphene quantum dots.

[0021] In another preferred example, the exogenous metal carbonyl (ML-CO) donor structure is [(tpy′)M(CO) n X]; where n is 2 or 3; X is selected from the group consisting of: Br - , Cl - , I - , H2O, CF3, SO3 - ;

[0022] where tpy′ is a tridentate nitrogen-containing ligand selected from 4'-formic acid-2,2':6',2”-terpyridine; and / or a bidentate nitrogen-containing ligand selected from 2,2′-bipyridine carboxylic acid derivatives, 2,9-phenanthroline carboxylic acid derivatives, or combinations thereof;

[0023] The exogenous metal M is a transition metal selected from Mn, Fe, Re, Ru, or combinations thereof.

[0024] In another preferred example, the [(tpy′)M(CO) n X] is (tpy COOH )Mn(CO)2Br.

[0025] In another preferred example, tpy′ contains at least one carboxylic acid group.

[0026] In another preferred example, tpy′ is 4'-formic acid-2,2':6',2”-terpyridine.

[0027] In another preferred example, the targeting group is triphenylphosphine (TPP) and its derivatives, preferably 4-[(triphenylphosphine)-methyl]-pyridine.

[0028] In another preferred example, the system further has one or more of the following characteristics:

[0029] (Z1) The weight ratio of the exogenous metal carbonyl (ML-CO) donor to the carrier is (0.2-0.5):1;

[0030] (Z2) The weight ratio of the iron chelator to the carrier is (0.1-0.3):1;

[0031] (Z3) The molar ratio of the exogenous metal carbonyl (ML-CO) donor to the targeting group is (1 ± 0.5):(1 ± 0.5).

[0032] In another preferred embodiment, the carrier is covalently linked to the exogenous metal carbonyl (ML-CO) donor.

[0033] In another preferred embodiment, the covalent bond is an amide bond.

[0034] In another preferred embodiment, the targeting group is coordinately linked to the exogenous metal carbonyl (ML-CO) donor.

[0035] In another preferred embodiment, the carrier is covalently linked to the iron chelator.

[0036] In a second aspect of the present invention, there is provided a method for preparing the nanocomposite drug system described in the first aspect of the present invention, comprising the steps of:

[0037] (i) Providing a targeting group, an exogenous metal carbonyl (ML-CO) donor, an iron chelator, and a carrier;

[0038] (ii) In an inert atmosphere and an inert solvent, in the presence of AgBF4, the targeting group is coordinately bound to the exogenous metal carbonyl (ML-CO) donor to form a composite molecule;

[0039] (iii) In the presence of a coupling agent, the composite molecule and the iron chelator are respectively linked to the carrier to form the nanocomposite drug system described in the first aspect of the present invention.

[0040] In another preferred embodiment, the targeting group is triphenylphosphine (TPP).

[0041] In another preferred embodiment, the targeting group is a triphenylphosphine derivative.

[0042] In another preferred embodiment, the coupling agent is selected from the group consisting of EDC, NHS, or a combination thereof, wherein EDC is 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride and NHS is N-hydroxysuccinimide.

[0043] In another preferred embodiment, step (ii) is carried out under light-shielded conditions.

[0044] In another preferred embodiment, in step (ii), the reaction temperature is 30 °C to 60 °C, preferably 40 °C - 50 °C.

[0045] In another preferred embodiment, in step (iii), the reaction temperature is 20 °C to 40 °C, preferably 25 °C - 35 °C.

[0046] In another preferred example, in step (i), the exogenous metal carbonyl (ML-CO) donor is [(tpy′)M(CO) n X], and it is prepared through the following steps:

[0047] (a1) Provide tpy′, where tpy′ is a tridentate nitrogen-containing ligand selected from 4'-formic acid-2,2':6',2”-terpyridine, and / or a bidentate nitrogen-containing ligand selected from 2,2′-bipyridine carboxylic acid derivatives, 2,9-phenanthroline carboxylic acid derivatives, or a combination thereof;

[0048] (a2) In an inert atmosphere and an inert solvent, the tpy′ reacts with M(CO) m X, where M is selected from Mn, Fe, Re, Ru; X is selected from the following group: Br - , Cl - , I - , H2O, CF3, SO3 - ; m is a positive integer from 2 to 6; thus obtaining the exogenous metal carbonyl (ML-CO) donor.

[0049] In another preferred example, the M(CO) m X is Mn(CO)5Br.

[0050] In another preferred example, in step (a2), the reaction temperature is 0°C to 45°C, preferably 10°C to 25°C.

[0051] In another preferred example, in step (a2), the reaction temperature is room temperature.

[0052] In another preferred example, the inert atmosphere is nitrogen and / or argon.

[0053] In another preferred example, the inert solvent is selected from the following group: C1-C6 alcohol solvents, C1-C6 hydrocarbon solvents, amide solvents, or a combination thereof.

[0054] In another preferred example, the inert solvent is selected from the following group: ethanol, dichloromethane, DMF, or a combination thereof.

[0055] In a third aspect of the present invention, there is provided a pharmaceutical composition, comprising: the nanocomposite drug system described in the first aspect of the present invention, and a pharmaceutically acceptable carrier.

[0056] In a fourth aspect of the present invention, there is provided a use of the pharmaceutical composition described in the third aspect of the present invention for preparing a drug for treating cancer.

[0057] In another preferred embodiment, the drug is a targeted anti-cancer drug, which combines a drug of the iron chelator type and an exogenous metal carbonyl (ML-CO) donor, achieving a synergistic anti-cancer effect.

[0058] In another preferred embodiment, the drug is used for anti-tumor treatment.

[0059] In another preferred embodiment, the nano-composite drug system can target specific sub-organelles of tumor cell mitochondria and can deliver CO by near-infrared light control and consume iron ions in cancer cells, performing combined administration of CO and iron chelator to achieve synergistic tumor treatment.

[0060] In a fifth aspect of the present invention, there is provided a method for combined administration of a nano-system, irradiating the nano-composite drug system described in the first aspect of the present invention with near-infrared light, so that the nano-composite system releases CO and iron chelator.

[0061] In a sixth aspect of the present invention, there is provided a method for treating cancer, the method comprising: administering to a subject in need a therapeutically effective amount of the nano-composite drug system described in the first aspect of the present invention, or a pharmaceutical composition thereof.

[0062] In another preferred embodiment, the dosage form of the pharmaceutical composition or preparation is an injection, tablet, capsule, pill, suspension or emulsion.

[0063] In a seventh aspect of the present invention, there is provided a multi-modal treatment method for tumors by a nano-drug system, irradiating the nano-composite drug system described in the first aspect of the present invention with near-infrared light, which can produce an obvious photothermal effect, thus realizing a multi-modal therapy for chemotherapy and photothermal therapy of tumors.

[0064] In another preferred embodiment, the wavelength of the near-infrared light is 808 nm.

[0065] In another preferred embodiment, the method is non-therapeutic and non-diagnostic.

[0066] In another preferred embodiment, the release is in vitro, irradiating the nano-composite drug system in solution with light, so as to release CO and at the same time produce a photothermal effect.

[0067] In another preferred embodiment, the irradiation mode is pulsed irradiation or continuous irradiation.

[0068] In another preferred embodiment, the light intensity of the irradiation is 100 - 2000 mW / cm².

[0069] In an eighth aspect of the present invention, there is provided a non-therapeutic method for targeting and delivering a nano-composite drug system to cells, comprising the steps of:

[0070] Incubate the nanocomposite drug system described in the first aspect of the present invention with the cells.

[0071] Wherein,

[0072] The lipophilic cationic triphenylphosphine targeting group in the nanocomposite drug system binds to the highly negatively charged inner mitochondrial membrane of the cells, so that the nanocomposite drug system is targeted and delivered to specific sub-organelles of the cells.

[0073] In another preferred example, the incubation conditions are 37±2 °C (preferably 37 °C), 24±1 hour.

[0074] In another preferred example, the concentration of the nanocomposite drug system is 5 - 500 μg / mL.

[0075] In the ninth aspect of the present invention, a method for co-delivering CO and iron chelator drugs is provided, including the step of administering the nanocomposite drug system described in the first aspect of the present invention to a subject in need.

[0076] In another preferred example, the subject includes mammals.

[0077] In another preferred example, the method further includes irradiating with near-infrared light for photocontrolled release of CO.

[0078] The multifunctional metal nano-antitumor drug system disclosed in the present invention uses an exogenous metal carbonyl (ML-CO) donor as an exogenous carbon monoxide donor, a chelating agent to consume iron ions in cancer cells, triphenylphosphine and its derivatives as targeting groups, and the targeting groups are connected to the exogenous metal carbonyl (ML-CO) donor through coordination bonds, and uses heteroatom-doped graphene quantum dots, etc. as carriers.

[0079] The metal nano-drug system of the present invention has fluorescence tracing, selectively targets specific cancer cells, can deliver carbon monoxide and iron chelators by near-infrared light photocontrol, and has a photothermal therapy function at the same time. The combined administration of carbon monoxide and iron chelators and the synergistic photothermal therapy achieve an efficient anti-cancer effect. This system has good biocompatibility and stability, and the present invention has potential application prospects and commercial value in the fields of combined drug administration and multi-modal treatment of tumors, etc.

[0080] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be elaborated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0081] Figure 1Schematic diagram of the co-loaded composite nano-drug system of carbon monoxide and iron chelator, showing that the system uses nanoparticles as carriers, and exogenous metal carbonyl (ML-CO) donors and iron chelators are connected to the carriers, and the exogenous metal carbonyl (ML-CO) donors are simultaneously connected to the targeting groups.

[0082] Figure 2 Transmission electron microscopy image and particle size distribution of the nano-system {DFX@TPPMnCO@N-GQDs}, showing that its particle size is 3-5 nm.

[0083] Figure 3 Shows the CO release effect of the optimized nano-system {DFX@TPPMnCO@N-GQDs} under near-infrared light irradiation. Among them, Figure A presents the detection of CO release in the nano-system by the hemoglobin method. Concentration: 1.0 μg / mL; Light intensity: 500-2000 mW / cm 2 , λ = 808 nm; Figure B presents the CO release rate of the nano-system under near-infrared light irradiation with different powers. Concentration: 1.0 μg / mL; Light intensity: 500-2000 mW / cm 2 , λ = 808 nm.

[0084] Figure 4 Shows the mitochondrial targeting delivery results of different nano-systems. Among them, the upper row is the confocal laser image of HeLa cells incubated with the nano-platform {DFX@TPPMnCO@N-GQDs} (15.0 μg / mL) at 37 °C for 4 hours; the lower row is the confocal laser image of HeLa cells incubated with the non-targeted control group {DFX@MnCO@N-GQDs} (15.0 μg / mL) at 37 °C for 4 hours.

[0085] Figure 5 Shows the cytotoxicity of the optimized nano-system {DFX@TPPMnCO@N-GQDs} and its control group. HeLa cells were co-incubated with the optimized nano-system {DFX@TPPMnCO@N-GQDs} and the control nano-system (30 μg / mL) respectively, irradiated under dark conditions and under 808 nm near-infrared light for 10 minutes, and then the cytotoxicity test results were carried out respectively. Specific implementation mode

[0086] After extensive and in-depth research, through a large number of screenings and tests, the present inventor has developed for the first time a novel composite nano-drug system in which an iron chelator is combined with an exogenous metal carbonyl (ML-CO) donor. This system can be targeted and delivered to specific sub-organelles within the mitochondria of tumor cells, effectively depleting iron ions in cancer cells, and can photochemically release a therapeutic amount of CO. The system includes a carrier and an exogenous metal carbonyl (ML-CO) donor and an iron chelator with mitochondrial targeting function that are connected to the carrier (such as covalently). The system of the present invention can not only rapidly release CO molecules under near-infrared light irradiation, but also regulate the release rate and release amount of CO within a wide range by adjusting the illumination time and illumination intensity. At the same time, the system can also deplete iron ions in cancer cells. In addition, the nano-composite drug system of the present invention also has the advantages of targeting, good biocompatibility and stability. On this basis, the present invention has been completed.

[0087] Term

[0088] As used herein, the terms "drug system of the present invention", "system of the present invention", "targeted photochemically CO-releasing nano-composite drug", "targeted photochemically CO-releasing nano-composite drug", "targeted photochemically CO-releasing nano-composite drug composition", and "nano-composite material drug system" are used interchangeably and all refer to a composition in which nanoparticles are used as a carrier and a mitochondrial-targeted exogenous metal carbonyl (ML-CO) donor and an iron chelator are connected to the carrier.

[0089] As used herein, the term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity.

[0090] As used herein, the term "compatible" means that the components in the composition can be mixed with the nano-composite drug system of the present invention and with each other without significantly reducing the efficacy of the active ingredient.

[0091] As used herein, the abbreviation "EDC" is 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride, and "NHS" is N-hydroxysuccinimide.

[0092] As used herein, the term "room temperature" refers to a temperature range of 0 - 45 °C, preferably 10 - 35 °C; more preferably 15 - 25 °C.

[0093] Nanoparticle carrier

[0094] The carrier applicable to the present invention is not particularly limited and can be various nanoparticle carriers commonly used in the art. Representative examples include, but are not limited to: graphene quantum dots, heteroatom- and / or metal ion-doped graphene quantum dots, carbon dots, heteroatom- or metal ion-doped carbon dots, titanium dioxide nanoparticles, upconversion nanoparticles, Fe3O4 magnetic nanoparticles, manganese dioxide nanoparticles, or combinations thereof.

[0095] Typically, the particle size of the nanoparticle carrier of the present invention is 1-100 nm, preferably 3-50 nm; more preferably, the particle size of the nanoparticle carrier is 3-8 nm.

[0096] As used herein, the nanoparticle carrier of the present invention is a surface-aminated nanoparticle carrier.

[0097] Exogenous metal carbonyl (ML-CO) donor

[0098] The exogenous metal carbonyl (ML-CO) donor applicable to the present invention is a transition metal carbonyl compound. A metal ruthenium nitrosyl complex can also be used as the gas donor for the release of nitric oxide in the present invention.

[0099] Typically, the exogenous metal carbonyl (ML-CO) donor is a metal manganese carbonyl complex or a metal iron carbonyl complex.

[0100] In the present invention, the weight ratio of the exogenous metal carbonyl (ML-CO) donor linked with a targeting group, an iron chelator, and the nanoparticles is 0.2-0.5:0.1-0.3:1.

[0101] Iron chelator

[0102] The iron chelator applicable to the present invention is not particularly limited, and an iron chelator containing a -COOH functional group is preferred. Representative examples include, but are not limited to: deferasirox (DFX), deferoxamine (DFO), gallic acid, 3,4-dihydroxybenzoic acid, etc.

[0103] Targeting group

[0104] The targeting group applicable to the present invention is not particularly limited and can be various targeting groups commonly used in the art. Representative examples include, but are not limited to: folic acid molecule, galactose molecule, triphenylphosphine and its derivatives, biotin, or combinations thereof.

[0105] In the present invention, the molar ratio of the targeting group and the exogenous metal carbonyl (ML-CO) donor is (1±0.5):(1±0.5), preferably 1:1.

[0106] Nanocomposite drug system

[0107] The nano-composite drug system of the present invention refers to composite particles with nanoparticles as carriers, on which mitochondrial-targeted exogenous metal carbonyl (ML-CO) donors and iron chelators are connected.

[0108] A simplified schematic diagram is as Figure 1 shown.

[0109] The composite drug system of the present invention can be used to deliver iron chelators and CO to cancer cell mitochondria, thereby being used for the combined treatment of cancer.

[0110] Preparation method

[0111] The present invention also provides a preparation method of the drug system described in the present invention, which generally includes the following steps:

[0112] (1) Provide a targeting group, an exogenous metal carbonyl (ML-CO) donor, an iron chelator, and a carrier;

[0113] (2) Coordinate the targeting group with the exogenous metal carbonyl (ML-CO) donor to obtain an exogenous metal carbonyl (ML-CO) donor connected with the targeting group, and load the exogenous metal carbonyl (ML-CO) donor connected with the targeting group and the iron chelator on the carrier, thereby forming the drug system as described in the first aspect.

[0114] In another preferred example, in step (1), the exogenous metal carbonyl (ML-CO) donor is a metal manganese carbonyl complex [(tpy′)Mn(CO)2Br], and tpy′ is 4'-formic acid-2,2':6',2”-terpyridine.

[0115] In another preferred example, in step (1), the iron chelator is deferasirox 4-[3,5-bis(2-hydroxyphenyl)-1,2,4-triazol-1-yl]benzoic acid.

[0116] In another preferred example, the nanoparticle carrier is a surface-aminated nanoparticle.

[0117] In another preferred example, the targeting group is 4-[(triphenylphosphine)-methyl]-pyridine.

[0118] Compared with the prior art, the main advantages of the present invention include:

[0119] (1) The nano-composite drug system of the present invention has the function of targeted drug delivery.

[0120] (2) The nano-composite drug system of the present invention can selectively deliver drugs to tumor cell mitochondria, and improve the anti-cancer effect by maximizing the damage to mitochondria.

[0121] (3) The present invention provides a method for combined controlled drug delivery using a nano-system. This system can photochemically release a therapeutic amount of CO, and at the same time, in combination with an iron chelator to deplete iron, it deprives cancer cells of the utilization of iron, significantly enhancing the anti-cancer effect of this system.

[0122] (4) The nano-composite drug system of the present invention has good biocompatibility and stability.

[0123] (5) The nano-composite material drug system of the present invention has blue autofluorescence and can monitor its distribution in cells.

[0124] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are generally carried out under conventional conditions (such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989)) or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight percentage and weight parts.

[0125] Unless otherwise defined, all professional and scientific terms used herein have the same meaning as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to the described content can be applied to the method of the present invention. The preferred methods and materials described herein are for illustrative purposes only.

[0126] Example 1

[0127] (1) Synthesis of the targeting group triphenylphosphine derivative 4-[(triphenylphosphine)-methyl]-pyridine (TPPPy)

[0128] Triphenylphosphine (6.48 g, 24.8 mmol) and 4-(methylchloro)pyridine hydrochloride (4.06 g, 24.8 mmol) were added to 50 mL of acetonitrile, and then heated under reflux for 24 h. After the reaction was completed and cooled to room temperature, the solid was separated by filtration, washed three times with ether, and dried in vacuo for 30 minutes to obtain an off-white intermediate. Under an ice-water bath condition, the obtained intermediate (9.19 g, 21.6 mmol) was dispersed in 40 mL of ether, and then potassium tert-butoxide (2.65 g, 23.8 mmol) was added to the above solution. The ice-water bath was removed and the reaction was continued to stir at room temperature for 4 hours, and then the solid was collected by filtration and washed three times with ether. The yellow solid was dissolved in 50 mL of water, and an aqueous solution containing NH4PF6 (3.86 g, 23.8 mmol) was added in an amount of 5 mL. After continuing to stir for 90 minutes, the precipitate was collected by filtration, washed with water and ether, and dried in vacuo to obtain 9.5 g of an off-white product with a yield of 87%.

[0129] (2) Synthesis of exogenous metal carbonyl (ML-CO) donor [(tpy COOH )Mn(CO)2Br]:

[0130] Add tpy COOH (110 mg, 0.4 mmol) and Mn(CO)5Br (120 mg, 0.44 mmol) into 10 mL of CH2Cl2. Wrap the round-bottom flask with tin foil and stir the mixed solution in the dark at room temperature for 12 h. After the reaction, collect the crude product by centrifugation and wash it three times with CH2Cl2 to remove the excess Mn(CO)5Br. Dry it at room temperature to obtain 188.8 mg of yellow product with a yield of 92.3%.

[0131] (3) Synthesis of CO donor TPPMnCO with mitochondrial targeting function:

[0132] Add tpy COOH MnCO (234 mg, 0.5 mmol), TPPPy (221 mg, 0.5 mmol) and AgBF4 (117 mg, 0.6 mmol) into 10 mL of DMF. Then wrap the resulting mixed solution with aluminum foil and reflux it in the dark at 50 °C for 6 h. After cooling to room temperature, remove the generated AgBr precipitate by centrifugation and add an excess of H2O to precipitate the product. Then wash it three times with water to remove the residual DMF and obtain 106.8 mg of yellow product after freeze-drying with a yield of 25.8%.

[0133] Example 2: Synthesis of nanoparticle composite drug system {DFX@TPPMnCO@N-GQDs}

[0134] (1) Preparation of N-GQDs@NH2 (surface amino-functionalized N-GQDs nanoparticles)

[0135] Weigh citric acid (40 mg), dissolve it in 40 mL of distilled water, add ammonia water (8 mL), and place the solution in a muffle furnace at 200 °C for 3 h. After cooling to room temperature, adjust the pH to 8 to obtain a light yellow solution. Dialyze the solution in water for 4 h using a dialysis bag with a molecular weight cut-off of 1000, and then rotary evaporate the solution to obtain N-GQDs solid. Dissolve N-GQDs in 2.0 mL of distilled water, add EDC / NHS, activate for 1 h, add 1 mL of anhydrous ethylenediamine, and stir at room temperature for 24 h. Dialyze the solution in water for 24 h using a dialysis bag with a molecular weight cut-off of 1000, and freeze-dry the solution inside the dialysis bag to obtain the product NH2@N-GQDs.

[0136] (2) Preparation of nanoparticle composite drug system {DFX@TPPMnCO@N-GQDs}

[0137] The iron chelator DFX (38 mg, 0.1 mmol) and the CO donor TPPMnCO with mitochondrial targeting function (92 mg, 0.1 mmol) were dissolved in 20 mL of DMF, and EDC / NHS was added at room temperature for 1 hour of activation. Then, 20 mL of an aqueous solution containing NH2@N-GQDs was added, and the mixed solution was stirred in the dark at room temperature for 24 hours. After the reaction, the mixture solution was dialyzed against deionized water in a dialysis bag (cut-off molecular weight 1000 Da) for 48 hours, and finally the nano-platform {DFX@TPPMnCO@N-GQDs} was obtained by freeze-drying.

[0138] The transmission electron micrograph and particle size distribution of the {DFX@TPPMnCO@N-GQDs} nano-drug system of this example are as Figure 2 shown, showing that its particle size is 3 - 8 nanometers.

[0139] Example 3: Detection of CO release outside cells

[0140] Detection of extracellular CO release by hemoglobin (Hb) method

[0141] Weigh 14.3 mg of hemoglobin and place it in a round-bottom flask. Then, add 50 mL of PBS buffer (10 mM, pH = 7.4) for dissolution, and deoxygenate the hemoglobin solution by bubbling with nitrogen. Subsequently, add 20 mg of sodium dithionite for reduction under a nitrogen atmosphere. Take 4.0 mL of the resulting reduced hemoglobin solution and slowly add it to a quartz cuvette with 4 mg of the nano-platform {DFX@TPPMnCO@N-GQDs} placed at the bottom, and then immediately seal it with a lid. Finally, record the ultraviolet-visible absorption spectrum at 350 - 600 nm after irradiation with 808 nm laser at different powers (0.2, 0.6, and 1.0 Wcm -2 ) and the results are as Figure 3 shown.

[0142] Results

[0143] It can be significantly seen that the release of extracellular CO by the nano-platform {DFX@TPPMnCO@N-GQDs} is positively correlated with the light intensity, and the stronger the light intensity, the more CO is released.

[0144] Example 4: Co-localization experiment of the {DFX@TPPMnCO@N-GQDs} nano-drug system in cancer cell mitochondria

[0145] After pre-incubating HeLa cells with the targeted nanomedicine {DFX@TPPMnCO@N-GQDs} or the control group without mitochondrial targeting group {DFX@MnCO@N-GQDs} (15 μg / mL) for 4 hours, the cells were washed twice with PBS buffer to remove the nanomaterials not phagocytosed by the cells, and then 1 mL of DMEM medium containing Mito-Tracker Green (100 nM) was added and incubated for another 30 minutes. Subsequently, the cells were washed three times with PBS buffer to remove the mitochondrial dye not phagocytosed by the cells and added to serum-free DMEM medium for imaging. Finally, excitation was carried out with lasers at λ = 405 and 488 nm, and the emission spectra in the wavelength ranges of 420 - 490 nm and 500 - 550 nm were collected respectively.

[0146] Results

[0147] As Figure 4 , the nanomedicine with mitochondrial targeting group {DFX@TPPMnCO@N-GQDs} can effectively accumulate in the subcellular mitochondria of HeLa cells.

[0148] Example 5: Toxicity test of {DFX@TPPMnCO@N-GQDs} nanomedicine system in cells

[0149] HeLa or 4T1 cells were seeded on 96-well plates at a cell density of approximately 5×10 4 , and incubated in a cell culture incubator at 37 °C with a humidified atmosphere containing 5% CO2 for 24 hours. After the cells adhered, the medium in the wells was removed, and serum-free cell culture medium containing different nanomaterials with a concentration of 30 μg / mL (carbonyl-free control sample {DFX@N-GQDs}, iron chelator-free control sample {TPPMnCO@N-GQDs}, targeting group-free control sample {DFX@MnCO@N-GQDs}, and targeted nanomedicine {DFX@TPPMnCO@N-GQDs}) was added and incubated for another 24 hours. After irradiation with an 808 nm laser (1.5 W / cm 2 ) for 10 minutes, incubation was continued for 4 hours. Subsequently, the medium was removed and the cells were washed with PBS buffer to remove the unphagocytosed materials. After measuring the background absorbance at 490 nm with a microplate reader, 100 μL of medium containing MTT (0.5 mg / mL) was added to each well. Incubation was continued for 3 - 4 hours. After removing the cell medium, 150 μL of DMSO was added to dissolve the formed formazan crystals, and finally the absorbance at 490 nm was measured with a microplate reader. The phototoxicity of the other control groups was also determined by the same procedure.

[0150] Results

[0151] As Figure 5, compared with other control groups, the nanoplatform {DFX@TPPMnCO@N-GQDs} with mitochondrial-targeted delivery of CO and iron ion chelation function showed the highest cytotoxicity to HeLa cells under light conditions.

[0152] All documents mentioned in this invention are cited in this application for reference as if each document was cited separately for reference. In addition, it should be understood that after reading the above teachings of this invention, those skilled in the art can make various changes or modifications to this invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

Claims

1. A nano-composite material drug system, characterized in that The structure of the described drug system is as follows: X-Y-Z-W Wherein, X represents an iron chelator; the iron chelator is selected from the group consisting of deferasirox 4-[3,5-bis(2-hydroxyphenyl)-1,2,4-triazol-1-yl]benzoic acid; Y represents a carrier, wherein the carrier is a surface-aminated nanoparticle; the nanoparticle is a surface-aminated nitrogen-doped graphene quantum dot; Z represents an exogenous metal carbonyl ML-CO donor; the exogenous metal carbonyl (ML-CO) donor has the structure [(tpy′)M(CO) n X]; wherein, n is 2 or 3; X is selected from the group consisting of: Br - , Cl - ; Wherein, tpy′ is a tridentate nitrogen-containing ligand selected from 4'-formic acid-2,2':6',2”-terpyridine; The exogenous metal M is a transition metal selected from Mn, Fe, Re, Ru, or a combination thereof; W represents a targeting group, and the targeting group is 4-[(triphenylphosphine)-methyl]-pyridine.

2. The system according to claim 1, wherein The particle size of the nanocomposite drug system is 3-50 nm.

3. The system according to claim 1, wherein The [(tpy′)M(CO) n X] is [(tpy′)Mn(CO)2Br].

4. The system according to claim 1, characterized in that, The system further has one or more of the following characteristics: (Z1) The weight ratio of the exogenous metal carbonyl (ML-CO) donor to the carrier is (0.2-0.5):1; (Z2) The weight ratio of the iron chelator to the carrier is (0.1-0.3):1; (Z3) The molar ratio of the exogenous metal carbonyl (ML-CO) donor to the targeting group is (1±0.5):(1±0.5).

5. The system according to claim 1, characterized in that, The carrier is covalently linked to the exogenous metal carbonyl (ML-CO) donor.

6. The system according to claim 1, characterized in that, The carrier is covalently linked to the iron chelator.

7. A preparation method of the nanocomposite drug system as described in claim 1, characterized in that, Comprising the steps of: (i) Providing a targeting group, an exogenous metal carbonyl (ML-CO) donor, an iron chelator, and a carrier; (ii) In an inert atmosphere and an inert solvent, in the presence of AgBF4, the targeting group is coordinatively bound to the exogenous metal carbonyl (ML-CO) donor to form a composite molecule; (iii) In the presence of a coupling agent, the composite molecule and the iron chelator are respectively linked to the carrier, thereby forming the nanocomposite drug system as claimed in claim 1; Wherein, the iron chelator is selected from the group consisting of deferasirox 4-[3,5-bis(2-hydroxyphenyl)-1,2,4-triazol-1-yl]benzoic acid; The carrier is a surface-aminated nitrogen-doped graphene quantum dot; The exogenous metal carbonyl (ML-CO) donor structure is [(tpy′)M(CO) n X]; where n is 2 or 3; X is selected from the group consisting of: Br - , Cl - ; Wherein, tpy′ is a tridentate nitrogen-containing ligand selected from 4'-formic acid-2,2':6',2”-terpyridine; The exogenous metal M is a transition metal selected from Mn, Fe, Re, Ru, or a combination thereof; The targeting group is 4-[(triphenylphosphine)-methyl]-pyridine.

8. The preparation method according to claim 7, characterized in that, In step (i), the exogenous metal carbonyl (ML-CO) donor is [(tpy′)M(CO) n X], and it is prepared through the following steps: (a1) Providing tpy′, wherein tpy′ is a tridentate nitrogen-containing ligand selected from 4'-formic acid-2,2':6',2”-terpyridine; (a2) In an inert atmosphere and an inert solvent, the said tpy′ reacts with M(CO) m X, where M is selected from Mn, Fe, Re, Ru; X is selected from the group consisting of: Br - , Cl - ; m is a positive integer from 2 to 6; thereby obtaining an exogenous metal carbonyl (ML-CO) donor.

9. The preparation method according to claim 8, characterized in that, The described M(CO) m X is Mn(CO)5Br.

10. A pharmaceutical composition, characterized in that, Comprising: The nanocomposite drug system as claimed in claim 1, and a pharmaceutically acceptable carrier.

11. Use of the pharmaceutical composition according to claim 10, characterized in that, For the preparation of a drug for treating cancer.

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