CN113509559B - CO and drug release synergistic therapeutic agent and preparation method and application thereof - Google Patents
CO and drug release synergistic therapeutic agent and preparation method and application thereof Download PDFInfo
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- CN113509559B CN113509559B CN202110345459.5A CN202110345459A CN113509559B CN 113509559 B CN113509559 B CN 113509559B CN 202110345459 A CN202110345459 A CN 202110345459A CN 113509559 B CN113509559 B CN 113509559B
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- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims abstract description 20
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- VSZWPYCFIRKVQL-UHFFFAOYSA-N selanylidenegallium;selenium Chemical compound [Se].[Se]=[Ga].[Se]=[Ga] VSZWPYCFIRKVQL-UHFFFAOYSA-N 0.000 description 1
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- JVBXVOWTABLYPX-UHFFFAOYSA-L sodium dithionite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])=O JVBXVOWTABLYPX-UHFFFAOYSA-L 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/26—Iron; Compounds thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
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- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
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- Polymers & Plastics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a CO and drug release synergistic therapeutic agent, and a preparation method and application thereof, and belongs to the technical field of medicines. The synergistic therapeutic agent comprises metal-organic framework nanoparticles Uio-66-SH-TPP and hyaluronic acid, wherein the hyaluronic acid is taken as a shell to wrap the outer surface of the metal-organic framework nanoparticles Uio-66-SH-TPP; the metal-organic framework nano-particles Uio-66-SH-TPP are modified with Fe through coordination bonds 3 (CO) 12 The medicine 5-FU is adsorbed in the pore canal of the metal-organic framework nano-particles Uio-66-SH-TPP. The synergistic therapeutic agent of the invention can realize the synergistic treatment of CO gas treatment and chemotherapy, thereby improving the accuracy of treating focal areas and enhancing the treatment effect.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a mitochondria-targeted CO and drug release synergistic therapeutic agent, and a preparation method and application thereof.
Background
Cancer is one of the most serious public health problems worldwide, and direct therapies such as chemotherapy, photodynamic therapy and photothermal therapy have been widely used for cancer treatment. However, these therapies all face a common problem of limited killing of cancer cells and toxic side effects on normal cells. These drawbacks seriously hamper the effective use of these therapies in cancer treatment. Although CO is a toxic gas, more and more researches indicate that the endogenous gas generated by heme metabolism is an important physiological gas signal molecule, plays an indispensable role in cytoprotection and maintaining the balance of the environment in cells, and can effectively regulate and control various signal paths in the cells, particularly the processes related to apoptosis and inflammatory reactions. However, CO and hemoglobin are very easy to combine, which makes CO a versatile problem for use as an inhaled medicament, such as dose control, targeted delivery, etc. Thus, the establishment of a targeted drug delivery system and the controlled release are key points for improving the CO gas treatment effect.
Chemotherapy remains one of the most important approaches to cancer treatment. However, the tumor microenvironment (Tumor microenvironment, TME) is often characterized by hypoxia, high hydrogen peroxide concentration, glucose deficiency and low pH, directly affecting the chemotherapeutic effect of cancer to a large extent. The chemotherapeutic pattern of a single anti-tumor drug may induce resistance and immunosuppressive effects in solid tumors, whereas the multiple chemotherapeutic pattern of a combination of multiple anti-tumor drugs appears to significantly enhance chemotherapy. Research shows that the gas treatment is an emerging new tumor treatment strategy with very good application prospect, and the synergistic chemotherapy can obviously enhance the effect of cancer treatment. Therefore, it is critical to design and construct a drug delivery system with targeted delivery and controlled release that is biocompatible.
Therefore, the method has the advantages of constructing a drug delivery system for targeting tumor cells, utilizing the characteristics of TME to stimulate response to CO gas and controllable release of drugs, realizing the effect of improving tumor treatment by gas treatment and cooperative chemotherapy, and having remarkable significance.
Disclosure of Invention
It is an object of the present invention to provide a TME-responsive CO and drug release CO-therapeutic agent.
It is another object of the present invention to provide a method for preparing the above-mentioned cotherapeutic agent and its use.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a CO and drug release CO-therapeutic agent comprising metal-organic framework nanoparticles Uio-66-SH-TPP and hyaluronic acid as an outer shell around the outer surface of the metal-organic framework nanoparticles Uio-66-SH-TPP;
the metal-organic framework nano-particles Uio-66-SH-TPP are modified with Fe through coordination bonds 3 (CO) 12 The medicine 5-FU is adsorbed in the pore canal of the metal-organic framework nano-particles Uio-66-SH-TPP.
The preparation method of the CO and drug release synergistic therapeutic agent comprises the following steps:
step 1, adding 2, 5-dimercapto terephthalic acid, (4-carboxybutyl) triphenylphosphine bromide, zirconium tetrachloride and glacial acetic acid into N, N' -dimethylformamide, and synthesizing Uio-66-SH-TPP by a solvothermal method;
step 2, fe 3 (CO) 12 Adding Uio-66-SH-TPP into tetrahydrofuran, and reflux reacting to obtain Fe modified material 3 (CO) 12 Uio-66-SH-TPP of (C);
step 3, fe is modified 3 (CO) 12 Uio-66-SH-TPP of (2) are dispersed in water, then 5-FU is added, and nano composite particles for adsorbing the 5-FU are obtained after stirring;
and 4, dispersing the nano composite particles for adsorbing the 5-FU in water, adding a PBS solution of hyaluronic acid, refrigerating the mixture in a refrigerator, taking out and centrifuging to obtain the TME-responsive CO and drug release synergistic therapeutic agent.
Further, in the step 1, a solvothermal method is adopted, and the mixture is placed in a polytetrafluoroethylene high-pressure reaction kettle for reaction for 24 hours at 120 ℃.
Further, the reflux reaction in the step 2 is to heat and condense at 70 ℃ for 1 hour.
Further, in the step 3, the stirring condition is 20-30 ℃ for 12 hours.
Further, the cooling condition in the step 4 is 4 ℃ for 2 hours.
The synergistic therapeutic agent obtained by the preparation method comprises Uio-66-SH-TPP nano particles with square structures and Fe with internal modification of a framework 3 (CO) 12 The material pore canal is internally adsorbed with 5-FU and the material biocompatibility surface is improved to coat HA.
The application of the synergistic therapeutic agent in preparing tumor therapeutic medicaments.
The beneficial effects are that: the invention is implemented by combining H 2 DMBD is used as an organic ligand of a metal-organic framework (MOFs) material, zirconium (Zr) is used as a metal source, a terminal hydroxyl (-OH) group of the Zr can be coordinated with a carboxyl (-COOH) group on the ligand, and a mitochondria targeting molecule TPP is introduced while synthesizing the MOFs; the organic ligand with mercapto (-SH) can be combined with Fe 3 (CO) 12 Introducing CO prodrug through coordination bond S-Fe, adsorbing medicine 5-FU in pore canal of porous nanometer MOFs composite material; HA is coated on the surface of the material, and a synergistic therapeutic agent with controllable CO and a drug release system is constructed. The therapeutic agent of the invention has the characteristics of targeted delivery and controllable release, and realizes the synergistic treatment of gas treatment and chemotherapy through TME stimulation response control. The invention adopts a step-by-step delivery mode of targeting tumor cell surfaces and organelles, and designs a drug delivery system capable of releasing CO and 5-FU by TME stimulation, thereby improving the defects of gas and drug leakage in the traditional treatment and enhancing the tumor treatment effect. In addition, the synthesis steps of the therapeutic agent are simpler, and the yield is higher; further, the synthesis method is simple and low in cost, so that the method is suitable for mass production.
Drawings
FIG. 1 is an SEM photograph (a), zeta potential (b), ultraviolet-visible-near infrared spectrum (c) and infrared spectrum (d) of Uio-66-SH-TPP nanoparticles in example 1.
FIG. 2 is a diagram of Uio-66-SH-TPP@Fe of example 1 3 (CO) 12 At 30 mu M H 2 O 2 And ferrous sulfate heptahydrate (namely 30 mu M. OH) by utilizing bovine hemoglobin to reduceUltraviolet spectrum time change curve (a) of the method and statistics (b) of CO release amount under PBS solution environments with different concentrations of OH.
FIG. 3 shows the ultraviolet spectrum (a) of 5-FU at various concentrations in example 1 and the standard curve (R 2 =0.9967) (b)。
FIG. 4 is an ultraviolet absorbance spectrum (a) of 5-FU in water at room temperature stirring load of 5-FU in water for different time intervals and a time variation-drug load curve (b) calculated from the graph of (a) of example 1, uio-66-SH-TPP.
FIG. 5 is a measurement of 5-FU release in different pH environments for Uio-66-SH-TPP@5-FU of example 1.
FIG. 6 is a graph showing fluorescence of drug and mitochondrial after mitochondrial staining of Uio-66-SH and Uio-66-SH-TPP (adsorbing rhodamine B capable of generating red fluorescence) in example 1 in culture with HeLa cells.
FIG. 7 shows the concentrations of Uio-66-SH-TPP, uio-66-SH-TPP@Fe in example 1 3 (CO) 12 、Uio-66-SH-TPP@ Fe 3 (CO) 12 @5-FU、Uio-66-SH-TPP@ Fe 3 (CO) 12 Cell Activity of @5-FU @ HA and 5-FU.
Detailed Description
In order to describe the structural features, technical means, and achieved objects and effects of the present invention in detail, the following description is made in connection with the embodiments and the accompanying drawings.
The invention provides a CO and drug release synergistic therapeutic agent for tumor microenvironment response, which comprises Uio-66-SH-TPP nano particles with mitochondrial targeting and Fe modified in a framework 3 (CO) 12 The 5-FU is adsorbed in the pore canal of the Uio-66-SH-TPP nano-particle, and meanwhile, the surface of the Uio-66-SH-TPP nano-particle is also coated with Hyaluronic Acid (HA) to further enhance the targeting of the drug delivery system.
Specifically, the Uio-66-SH-TPP nanoparticle is a square structure with a diameter of about 20 nm.
Ligand 2, 5-dimercapto terephthalic acid (H) used in the present invention 2 DMBD), which can be prepared by the following procedure, or can be purchased directly from commercial sources.
2, 5-dimercapto terephthalic acid (H) 2 DMBD), comprising the steps of:
(1) Synthesis of diethyl 2, 5-bis (methylthioaminomethoxy) terephthalate
5g of diethyl 2, 5-dihydroxyterephthalate and 9g of triethylenediamine (DABCO) were dissolved in 50mL of N, N-Dimethylacetamide (DMA) and ice-cooled to 0 ℃; 9.5g of N, N-dimethylthiocarboxychloride was dissolved in 25mL of DMA and after complete dissolution by sonication in nitrogen (N 2 ) Slowly dripping the solution into the solution in the previous step by using a constant-pressure dropping funnel under the protection, and keeping the temperature at 0 ℃; after the completion of the dropwise addition, the mixture was stirred at room temperature for 16 hours to give a white precipitate, which was suction-filtered and washed with water in a large amount and dried under vacuum to give the compound diethyl 2, 5-bis (methylthioaminomethoxy) terephthalate.
(2) Synthesis of diethyl 2, 5-bis (methylthio-sulfamoyl) terephthalate
At N 2 Under the protection, the synthesized diethyl 2, 5-bis (methylthioamino methoxy) terephthalate is heated and stirred for 1h at 230 ℃, the obtained brown mixture is slowly cooled to 70 ℃, 20mL of ethanol is added by a constant pressure dropping funnel, light brown crystals are obtained after the slow cooling to room temperature, the light brown crystals are dried after suction filtration, and finally the compound is purified by a column to obtain the diethyl 2, 5-bis (methylthioamino methylsulfonyl) terephthalate.
(3) Synthesis of 2, 5-dimercapto terephthalic acid
1.3 mol.L of preparation -1 KOH ethanol/water (1:1) solution and 20mL were degassed for 1h; the diethyl 2, 5-bis (methylthio-sulfamoyl) terephthalate obtained by the reaction was dissolved in degassed KOH ethanol/water (1:1) solution in N 2 Reflux is carried out for 3 hours at the temperature of 85 ℃ under protection; after the reaction mixture was cooled to room temperature, 10mL of concentrated hydrochloric acid was added in an ice bath to give a bright yellow precipitate, which was suction-filtered, washed with a large amount of water and dried in vacuo to give ligand 2, 5-dimercapto terephthalic acid (H) 2 DMBD）。
The preparation method of the synergistic therapeutic agent comprises the following steps:
step 1, synthesizing Uio-66-SH-TPP by adopting a solvothermal method;
9.6mg of zirconium tetrachloride (ZrCl) 4 ) 9.5mg of H 2 DMBD, 5mg of (4-carboxybutyl) triphenylphosphine bromide (TPP) and 750 mu L of glacial acetic acid are added into 10mL of N, N-Dimethylformamide (DMF), and the mixture is transferred into a polytetrafluoroethylene high-pressure reaction kettle for reaction at 120 ℃ for 24 hours after ultrasonic dispersion. After the reaction is cooled to room temperature, the mixture is centrifugally washed for 3 times by DMF and then centrifugally washed for 3 times by absolute ethyl alcohol, and then Uio-66-SH-TPP is obtained by vacuum drying.
The method for preparing Uio-66-SH is similar to the method for preparing Uio-66-SH-TPP, and only the raw material TPP is removed.
The optimum temperature for the reaction was 120 ℃ and the reaction time was 24h. The diameter of the obtained Uio-66-SH-TPP nanoparticle is about 20 nm.
Step 2, adopting coordination bonding to prepare Uio-66-SH-TPP@Fe 3 (CO) 12 ；
25mg of Uio-66-SH-TPP, 50 mg of Fe 3 (CO) 12 And 50mL in a round bottom flask, ultrasonic dispersion, the mixture was heated at 70 ℃ to condense and reflux for 1h. Finally, collecting the product by centrifugation, centrifugally washing the product with absolute ethyl alcohol for 3 times, and vacuum drying to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 。
The 2, 5-dihydroxyterephthalic acid diethyl ester, triethylene diamine, N, N-dimethyl thiocarboxychloride and ZrCl 4 KOH, DMA, DMF, etc. are all common chemical raw materials and 5-FU and HA used in the next step, and can be directly ordered from a reagent net.
Step 3, adsorbing 5-FU to Uio-66-SH-TPP@Fe by adopting a normal-temperature stirring method 3 (CO) 12 The nanometer particle pore canal;
5.5mg of Uio-66-SH-TPP@Fe 3 (CO) 12 Dispersing in 10mL PBS, adding 14mg of 5-FU, magnetically stirring at room temperature overnight, centrifuging, collecting, and washing with water 3 times to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU。
Method for preparing Uio-66-SH-TPP@5-FU is the same as for preparing Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FUThe method is similar, and the raw materials Uio-66-SH-TPP@Fe are only used 3 (CO) 12 And changing into Uio-66-SH-TPP.
The method for preparing Uio-66-SH and Uio-66-SH-TPP adsorption Rhodamine B (RB) capable of generating red fluorescence is similar to the method for preparing Uio-66-SH-TPP@5-FU, and only the raw material 5-FU is changed into RB.
Step 4, preparing Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU@HA；
15mg of Uio-66-SH-TPP@Fe 3 (CO) 12 The @5-FU was dispersed in 4mL of water and 15mg of Hyaluronic Acid (HA) (dissolved in 6mL of PBS solution) was gradually added. Refrigerating the mixture in a 4 deg.C refrigerator for 2 hr, centrifuging, collecting, and washing with water for 2 times to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU@HA。
The synergistic therapeutic agent obtained by the preparation method comprises Uio-66-SH-TPP nano particles with square structures and Fe with internal modification of a framework 3 (CO) 12 The material pore canal is internally adsorbed with 5-FU and the material biocompatibility surface is improved to coat HA.
The Uio-66-SH-TPP nano particles have a diameter of about 20nm and Fe is introduced into the skeleton 3 (CO) 12 And (3) functional modification of the pore canal adsorption 5-FU and the surface coating HA.
The synergistic therapeutic agent of the invention has the synergistic therapeutic function of CO gas treatment and chemotherapy, and Uio-66-SH-TPP@Fe of the invention 3 (CO) 12 Mitochondrial targeting and intra-mitochondrial OH triggers CO release. The 5-FU contained in the co-therapeutic nanomaterial may stimulate a responsive release to effect chemotherapy using a low pH in TME. The HA coated on the surface of the material can realize the purpose of tumor targeted drug delivery through a ligand-receptor binding mechanism with a receptor overexpressed on the surface of a tumor cell, and TPP targets the mitochondria of the tumor cell. Thus, the step-by-step targeting effect of cell targeting and organelle (mitochondria) targeting can be realized, and the targeting of the cooperative therapeutic agent is enhanced. Therefore, the constructed drug delivery system can realize the CO-treatment of CO gas treatment and chemotherapy, thereby improving the accuracy of treating the focus area and enhancing the treatment effect.
The prepared synergistic therapeutic agent can be applied to the preparation for treating tumors.
The preparation for treating tumor is gas treatment cooperative chemotherapy of the cooperative therapeutic agent under the condition of stimulating response to CO gas and drug release.
It will be appreciated that the co-therapy is capable of inhibiting the growth of tumour cells and killing cancer cells and therefore the co-therapeutic agent of the present invention is a highly effective, low toxic, green form of tumour therapy.
The preparation method of the tumor synergistic therapeutic agent has the advantages of low cost of synthetic raw materials, simple preparation process and easy mass production. In addition, the synergistic therapeutic agent obtained by the preparation method has good monodispersity and stability, good biocompatibility, high tumor targeting and controllable CO and 5-FU release.
The invention is further illustrated by the following examples.
Example 1
(1) Preparation of ligand H 2 DMBD
A. Synthesis of diethyl 2, 5-bis (methylthioaminomethoxy) terephthalate
5g of diethyl 2, 5-dihydroxyterephthalate and 9g of DABCO were dissolved in 50mL of DMA and cooled to 0℃in an ice bath; 9.5g of N, N-dimethylthiocarboxychloride was dissolved in 25mL of DMA and sonicated to dissolve completely in N 2 Slowly dripping the solution into the solution in the previous step by using a constant-pressure dropping funnel under the protection, and keeping the temperature at 0 ℃; after the completion of the dropwise addition, the mixture was stirred at room temperature for 16 hours to give a white precipitate, which was suction-filtered and washed with water in a large amount and dried under vacuum to give the compound diethyl 2, 5-bis (methylthioaminomethoxy) terephthalate.
B. Synthesis of diethyl 2, 5-bis (methylthio-sulfamoyl) terephthalate
At N 2 Under protection, the synthesized diethyl 2, 5-bis (methylthioaminomethoxy) terephthalate was heated and stirred at 230℃for 1h, the resulting brown mixture was slowly cooled to 70℃and 20mL of ethanol was added using a constant pressure dropping funnel and slowly cooledLight brown crystals are obtained at room temperature, the light brown crystals are dried after suction filtration, and finally the compound is purified by a column to obtain the diethyl 2, 5-bis (methylthio-amino-methylsulfonyl) terephthalate.
C. Synthesis of 2, 5-dimercapto terephthalic acid
1.3 mol.L of preparation -1 KOH ethanol/water (1:1) solution and 20mL were degassed for 1h; the diethyl 2, 5-bis (methylthio-sulfamoyl) terephthalate obtained by the reaction was dissolved in degassed KOH ethanol/water (1:1) solution in N 2 Reflux is carried out for 3 hours at the temperature of 85 ℃ under protection; after the reaction mixture was cooled to room temperature, 10mL of concentrated hydrochloric acid was added in an ice bath to give a bright yellow precipitate, which was suction-filtered, washed with a large amount of water and dried in vacuo to give the compound 2, 5-dimercapto terephthalic acid.
(2) Uio-66-SH-TPP nanoparticles;
9.6mg of ZrCl 4 9.5mg of H 2 DMBD, 5mg of TPP and 750 mu L of glacial acetic acid are added into 10mLDMF, and the mixture is transferred into a polytetrafluoroethylene high-pressure reaction kettle for reaction for 24 hours at 120 ℃. After the reaction is cooled to room temperature, the mixture is centrifugally washed for 3 times by DMF and then centrifugally washed for 3 times by absolute ethyl alcohol, and then Uio-66-SH-TPP is obtained by vacuum drying.
The method for preparing Uio-66-SH is similar to the method for preparing Uio-66-SH-TPP, and only the raw material TPP is removed.
(3) Preparation of Uio-66-SH-TPP@Fe 3 (CO) 12 ;
25mg of Uio-66-SH-TPP, 50 mg of Fe 3 (CO) 12 And 50mL THF was added to the round bottom flask and after ultrasonic dispersion, the mixture was heated to reflux at 70 ℃ for 1h. Finally, collecting the product by centrifugation, centrifugally washing the product with absolute ethyl alcohol for 3 times, and vacuum drying to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 。
(4) Preparation of Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU;
5.5mg of Uio-66-SH-TPP@Fe 3 (CO) 12 Dispersing in 10mL PBS, adding 14mg of 5-FU, magnetically stirring at room temperature overnight, centrifuging, collecting, and washing with water 3 times to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU。
Method for preparing Uio-66-SH-TPP@5-FU is the same as for preparing Uio-66-SH-TPP@Fe 3 (CO) 12 The method of @5-FU is similar, and only the raw materials Uio-66-SH-TPP @ Fe 3 (CO) 12 And changing into Uio-66-SH-TPP.
The method for preparing Uio-66-SH and Uio-66-SH-TPP adsorption Rhodamine B (RB) capable of generating red fluorescence is similar to the method for preparing Uio-66-SH-TPP@5-FU, and only the raw material 5-FU is changed into RB.
(5) Preparation of Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU@HA;
15mg of Uio-66-SH-TPP@Fe 3 (CO) 12 The @5-FU was dispersed in 4mL of water and 15mg of HA (dissolved in 6mL of PBS solution) was gradually added. The mixture is put into a refrigerator for refrigeration for 2 hours, and then centrifugally collected and washed with water for 2 times to obtain Uio-66-SH-TPP@Fe 3 (CO) 12 @5-FU@HA。
Performance test:
1. topography determination of Uio-66-SH-TPP nanoparticles
FIG. 1 is SEM (a), zeta potential (b), infrared spectrum (c) and ultraviolet-visible-near infrared spectrum (d) of Uio-66-SH-TPP nanoparticles prepared in example 1. As a result, it was found that the synthesized Uio-66-SH-TPP nanoparticle had a diameter of about 20nm in the graph (a). FIG. (b) shows that Uio-66-SH-TPP@Fe was prepared 3 (CO) 12 The absolute value of the Zeta potential of the material ratio Uio-66-SH-TPP nano particles is larger, which shows that the stability is better. It can be seen from the graph (c) that the prepared Uio-66-SH-TPP nanoparticle has excellent light absorption in the vacuum ultraviolet region. FIG. (d) shows that Uio-66-SH-TPP@Fe was prepared 3 (CO) 12 Uio-66-SH-TPP and Fe 3 (CO) 12 Is a characteristic peak of (2).
2. Determination of Uio-66-SH-TPP@Fe 3 (CO) 12 CO release in an OH environment
The CO release was detected spectrophotometrically by conversion of bovine hemoglobin (Hb) to CO followed by carbonyl hemoglobin (HbCO) with an ultraviolet absorption peak from 430nm (Hb absorption peak) to 410nm (HbCO absorption peak).
First, uio-66-SH-TPP@Fe prepared in example 1 3 (CO) 12 Preparing 100 [ mu ] g/mL PBS solution, dissolving freshly prepared Hb (4.2 [ mu ] M) in PBS (pH=7.4) solution, adding 1.2mg sodium dithionite to reduce the solution, adding 75 [ mu ] L of material with prepared concentration, and adding the solution into solutions (30 [ mu ] M,10 [ mu ] M,0 [ mu ] M H 2 O 2 And ferrous sulfate heptahydrate in PBS, ph=7.4). The entire reaction solution (3 mL) was immediately sealed in a 4mL ultraviolet quartz tube. Ultraviolet absorption spectrum of solutionI=350-600 nm) was collected on an ultraviolet/visible spectrophotometer. To eliminate influencing factors and to improve accuracy, due to HbCO and Hb, respectivelyITwo strong adsorption bands =410 and 430nm were used to quantify Hb to HbCO conversion to give fig. 2 (a), then by the calculation formula(Cco represents the concentration of CO,I 410nm represents HbCO inIUv absorbance value at 410nm,I 430nm represents Hb inI=430 and nm uv absorbance value) to obtain Uio-66-SH-tpp@fe 3 (CO) 12 CO release profile in 30. Mu.M.OH PBS solution 2 (b). From FIG. 2 (b), it can be seen that Uio-66-SH-TPP@Fe 3 (CO) 12 The amount of CO released at different concentrations of OH increases with increasing concentration of OH.
3. Drawing a standard curve of 5-FU
Aqueous solutions of 5-FU at different concentrations were prepared at 0.002mg/mL, 0.004mg/mL, 0.006mg/mL, 0.008mg/mL, 0.010mg/mL, 0.012mg/mL, 0.014mg/mL, 0.016mg/mL, 0.018mg/mL and 0.020mg/mL, respectively. The ultraviolet absorption spectra of 5-FU at different concentrations were measured, and it can be seen from FIG. 3 (a) that 5-FU has a maximum absorption peak at 262nm, and the maximum absorption peak increases with increasing concentration. A standard curve is drawn according to the corresponding ultraviolet absorption values of different concentrations at 262nm, and as can be seen from FIG. 3 (b), there is a good linear relationship between the two (R 2 =0.9967), it is also possible to obtain the concentration (X) -absorbance (Y) as a function: y=47.44 x-0.003867.
4. Determination of drug load of Uio-66-SH-TPP@5-FU
5.5mg of Uio-66-SH-TPP@Fe 3 (CO) 12 Disperse in 10mL PBS, add 14mg of 5-FU magnetically stir at room temperature. At time intervals of 0.5h, 1.0h, 1.5h, 2.0h, 3.0h, 4.0h, 5.0h, 6.0h, 7.0h, 8.0h, 9.0h, 10h, 11h and 12h, 3mL of the reaction solution was taken, centrifuged, and the supernatant was taken for measuring ultraviolet absorption spectrum. As can be seen from FIG. 4 (a), the absorbance value of the supernatant was continuously decreased with the increase of the reaction time, indicating that 5-FU was adsorbed into the pores of the material. According to the standard curve of 5-FU, the unadsorbed 5-FU in the supernatant of different reaction times is calculated, the drug load of different times is indirectly calculated, the change of the load is shown in figure 4 (b), and as the reaction time is prolonged, the 5-FU is gradually adsorbed into the material and reaches adsorption stability.
5. Determination of drug Release of Uio-66-SH-TPP@5-FU under different pH Environment
4mg/mL of the Uio-66-SH-TPP@5-FU aqueous solution prepared in example 1 was added with 10mL of PBS solutions prepared to have pH values of 4.6, 5.5, 6.5 and 7.4 respectively, 3mL of the reaction solution was taken at time intervals of 2.0h, 4.0h, 6.0h, 8.0h, 10h, 12h and 14h, and the supernatant was centrifuged to measure ultraviolet absorption spectrum. The release of 5-FU from the supernatant of different reaction times was calculated based on the standard curve of 5-FU, and the release of drug at different times for each pH was calculated, the change in release being shown in FIG. 5, which shows that as the pH was decreased, the release of 5-FU increased, indicating that 5-FU stimulated release under acidic conditions.
6. Determination of mitochondrial targeting of Uio-66-SH and Uio-66-SH-TPP
Drug fluorescence and mitochondrial fluorescence pictures after mitochondrial staining in culture with HeLa cells
Uio-66-SH and Uio-66-SH-TPP (after adsorption of RB) prepared in example 1 were formulated as 320 μg/mL (pH=7.4) PBS solution, heLa cells were treated at 10 5 Density of cells/well 2 phi 20mm glass bottom cell culture dishes were inoculated at 5% CO 2 Incubate at 37 ℃ for 12h. Old culture medium is removed, 1.5mL of fresh culture medium is added, and 500 mu m of fresh culture medium is added respectivelyL formulated material at 5% CO 2 Incubate at 37 ℃ for 4h. Cell dishes were washed with PBS to remove non-ingested particles, 1.5mL fresh medium and 500 μl mitochondrial stain were added at 5% CO 2 Incubate at 37 ℃ for 45 minutes. The cell dishes were washed with PBS to remove non-ingested stain, 500 μl of 4% paraformaldehyde cell fixative was added, after 20 min fixation, the cell dishes were washed with PBS to remove excess fixative, and after 1ml of each was added, both mitochondrial targeting was observed with a fluorescence microscope. From fig. 6, it can be seen that red fluorescence represents the position of the material, green fluorescence represents the position of the mitochondria, and from the fluorescence superposition diagram of the two, uio-66-SH is mostly red, while the Uio-66-SH-TPP has a plurality of yellow colors with red and green superposed, which indicates that Uio-66-SH-TPP targets mitochondria more strongly than Uio-66-SH.
7. Materials Uio-66-SH-TPP, uio-66-SH-TPP@Fe with different concentrations 3 (CO) 12 、Uio-66-SH-TPP@ Fe 3 (CO) 12 @5-FU、Uio-66-SH-TPP@ Fe 3 (CO) 12 Cellular Activity of @5-FU@HA and 5-FU
Cell viability of Hela cells was determined by using the thiazole blue (MTT) assay. The cells were packed in 10 4 Density of individual/well was inoculated into 96-well cell culture plates and incubated at 5% CO 2 Incubate at 37 ℃ for 12h. Then, the dosing group was dosed at 50. Mu.L per well of Uio-66-SH-TPP, uio-66-SH-TPP@Fe 3 (CO) 12 、Uio-66-SH-TPP@ Fe 3 (CO) 12 @5-FU、Uio-66-SH-TPP@ Fe 3 (CO) 12 The @5-FU @ HA and 5-FU were dispersed in DMEM and different concentrations (5, 10, 20, 40, 80 and 160 μg/mL) were added to each well. Cells were incubated at 5% CO 2 Incubate at 37 ℃ for an additional 24h. After incubation, old medium was removed and the cell wells were washed with PBS to remove non-ingested particles, then 100 μl of fresh medium was added. 10 μl of filter sterilized MTT reagent (5 mg/mL in PBS) was then added to each well and the plates incubated at 37 ℃. After an additional 4h incubation, the medium was removed and the precipitated formazan crystals were dissolved by addition of DMSO. The absorbance of the dissolved formazan crystals in each well was measured at 450nm using an enzyme-labeled instrument. With untreated cells as controlsThe cell viability was calculated for each concentration in the group, with the cell activity recorded as 100%. All samples were prepared in triplicate.
As can be seen from the cytotoxicity results of FIG. 7, the cell activity of Uio-66-SH-TPP is more than 97%, which indicates that the cytotoxicity is small and the biocompatibility of the material is good; uio-66-SH-TPP@Fe 3 (CO) 12 Cell activity lower than Uio-66-SH-TPP due to the introduced Fe 3 (CO) 12 Can target mitochondria and release CO, and generate gas treatment to reduce cell activity; uio-66-SH-TPP@Fe 3 (CO) 12 Ratio of @5-FU Uio-66-SH-TPP @ Fe 3 (CO) 12 The reason for the low cell activity of the drug 5-FU loaded on the material under the same condition is that the cell activity is reduced by the stimulated release of TME, uio-66-SH-TPP@Fe 3 (CO) 12 The cellular activity of @5-FU is lower than that of 5-FU because of the Fe introduced by the material under the same conditions 3 (CO) 12 CO release can be triggered by intra-mitochondrial OH, generating gas therapy to reduce cellular activity; uio-66-SH-TPP@Fe 3 (CO) 12 The @5-FU @ HA is lower than Uio-66-SH-TPP @ Fe 3 (CO) 12 Cell Activity of @5-FU because Uio-66-SH-TPP @ Fe 3 (CO) 12 The HA introduced by the @5-FU@HA can lead the material to target the surface of cells, further enhance the targeting of the drug delivery system, reduce the toxic and side effects of the drug delivery system on normal cells, lead the drug delivery system to have stronger purpose by a step-by-step targeting mode of cell-mitochondria, not kill the cells indiscriminately and further reduce the activity of tumor cells.
The synergistic therapeutic agent provided by the invention takes Uio-66-SH-TPP of modified mitochondrial targeting molecule TPP as a carrier and modifies Fe in the framework 3 (CO) 12 The targeting of the drug delivery system is further enhanced by adsorbing the drug 5-FU in the pore canal and coating HA on the surface. The nanometer MOFs-Uio-66-SH-TPP is prepared by a high-temperature solvothermal method, and has a small size of 20 nm. Secondly, fe is bonded by coordination bonding 3 (CO) 12 Modified into the Uio-66-SH-TPP skeleton to be used as a CO gas release prodrug. Finally, further modifying 5-FU and HA, triggering CO release by utilizing intra-mitochondrial OH, stimulating 5-FU release in TME,finally, the CO-therapeutic effect of CO gas treatment and chemotherapy is realized.
Claims (1)
1. The application of the CO and drug release synergistic therapeutic agent in preparing the tumor therapeutic drug is characterized in that,
the CO and drug release CO-therapeutic agent comprises metal-organic framework nanoparticles Uio-66-SH-TPP and hyaluronic acid as a shell coated on the outer surface of the metal-organic framework nanoparticles Uio-66-SH-TPP; the metal-organic framework nano-particles Uio-66-SH-TPP are modified with Fe through coordination bonds 3 (CO) 12 The medicine 5-FU is adsorbed in the pore canal of the metal-organic framework nano particles Uio-66-SH-TPP;
the CO and drug release CO-therapeutic agent is prepared by the steps of:
step 1, adding 2, 5-dimercapto terephthalic acid, (4-carboxybutyl) triphenylphosphine bromide, zirconium tetrachloride and glacial acetic acid into N, N' -dimethylformamide, and synthesizing Uio-66-SH-TPP by a solvothermal method;
step 2, fe 3 (CO) 12 Adding Uio-66-SH-TPP into tetrahydrofuran, and reflux reacting to obtain Fe modified material 3 (CO) 12 Uio-66-SH-TPP of (C);
step 3, fe is modified 3 (CO) 12 Uio-66-SH-TPP of (2) are dispersed in water, then 5-FU is added, and nano composite particles for adsorbing the 5-FU are obtained after stirring;
step 4, dispersing the nano composite particles for adsorbing the 5-FU in water, adding a PBS solution of hyaluronic acid, refrigerating the mixture in a refrigerator, taking out and centrifuging to obtain the TME-responsive CO and drug release synergistic therapeutic agent;
in the step 1, a solvothermal method is adopted, and the mixture is placed in a polytetrafluoroethylene high-pressure reaction kettle to react for 24 hours at 120 ℃;
in the step 2, the reflux reaction is heating, condensing and refluxing for 1h at 70 ℃;
in the step 3, stirring conditions are 20-30 ℃ for 12 hours;
and 4, cooling at 4 ℃ for 2 hours.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106478959A (en) * | 2016-09-23 | 2017-03-08 | 肇庆学院 | A kind of Cd MOF Ji Yu 4,4 ' dicarboxydiphenyl ether and preparation method thereof |
| CN108837159A (en) * | 2018-07-17 | 2018-11-20 | 西北农林科技大学 | A kind of nano antibacterial agent and preparation method thereof |
| CN112121154A (en) * | 2020-10-20 | 2020-12-25 | 南通大学 | Tumor microenvironment response CO gas therapeutic agent and preparation and application thereof |
| CN112210084A (en) * | 2020-10-27 | 2021-01-12 | 洪月恒 | Preparation method of metal organic framework with photocatalytic reduction of carbon dioxide |
| CN113144175A (en) * | 2021-03-31 | 2021-07-23 | 南通大学 | Tumor microenvironment response CO gas therapeutic agent and preparation method and application thereof |
-
2021
- 2021-03-31 CN CN202110345459.5A patent/CN113509559B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106478959A (en) * | 2016-09-23 | 2017-03-08 | 肇庆学院 | A kind of Cd MOF Ji Yu 4,4 ' dicarboxydiphenyl ether and preparation method thereof |
| CN108837159A (en) * | 2018-07-17 | 2018-11-20 | 西北农林科技大学 | A kind of nano antibacterial agent and preparation method thereof |
| CN112121154A (en) * | 2020-10-20 | 2020-12-25 | 南通大学 | Tumor microenvironment response CO gas therapeutic agent and preparation and application thereof |
| CN112210084A (en) * | 2020-10-27 | 2021-01-12 | 洪月恒 | Preparation method of metal organic framework with photocatalytic reduction of carbon dioxide |
| CN113144175A (en) * | 2021-03-31 | 2021-07-23 | 南通大学 | Tumor microenvironment response CO gas therapeutic agent and preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| "Enhanced Photochemical Hydrogen Production by a Molecular Diiron Catalyst Incorporated into a Metal−Organic Framework";Sonja Pullen et al.;《J. Am. Chem. Soc. 》;第第2013卷卷(第第135期期);第16997−17003页 * |
| Recent advances in the development of metal– organic framework-based gas-releasing nanoplatforms for synergistic cancer therapy;Danni Jin et al.;《Dalton Trans.》;20201217;第第2021卷卷(第第50期期);第1189–1196页 * |
| 线粒体靶向聚合物TPP-PEI-LND的构建及其特性;张兵锋;姜虎林;;中国药科大学学报(第06期);第659-664页 * |
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