Drug-loaded ligand, chromium-based metal organic framework material as well as preparation method and application of chromium-based metal organic framework material

Abstract

The invention is applicable to the technical field of materials, and provides a drug-loaded ligand, a chromium-based metal organic framework material and a preparation method and application thereof, and the preparation method of the drug-loaded ligand comprises the following steps: stirring cysteine methyl ester, HATU, 2, 5-di (methoxyformyl) benzoic acid and diisopropylethylamine in DMF at room temperature for 12 hours, washing and purifying; 3 mL of a sodium hydroxide aqueous solution is added into the obtained 2-((3-sulfydryl-1-methoxy-1-oxopropyl-2-yl) carbamoyl) dimethyl terephthalate in THF / MeOH, stirring is carried out for 18 h at the room temperature, acidification and reduced pressure concentrated acid treatment are carried out, and the drug-loaded ligand is obtained. The metal organic framework nanoparticles are prepared by adopting the drug-loaded ligand with the cysteine active group, the traditional physical adsorption or surface modification is broken through, and meanwhile, a functional group with radiation protection capability is bonded to the ligand through a ligand doping technology, so that drug administration is realized.

Description

Technical Field

[0001] The present application belongs to the field of material technology, and in particular relates to a drug-carrying ligand, a chromium-based metal organic framework material, and a preparation method and application thereof. Background Art

[0002] Drug delivery systems based on nanomaterials have become a good alternative to traditional methods, which can reduce side effects and improve efficacy. Metal organic frameworks (MOFs) have many potential advantages over other materials, such as diverse structures, high specific surface area, large pore size for drug loading, and inherent biodegradability. In 2005, Ferey's group first reported that MOF series metal organic framework materials were used as potential drug carriers. Since then, research on MOF as a drug carrier has continued to emerge. With the continuous development of nuclear application technology in the world, especially the emergence of nuclear power plants and radiation processing technology, the occurrence of radiation accidents has shown an increasing trend. On the other hand, the number of cancer patients receiving radiotherapy has also increased year by year. Therefore, the research on radiation protection drugs is not only a national nuclear safety demand, but also closely related to people's daily life. It is a major medical issue and should be given enough attention. Ionizing radiation can produce reactive oxygen species (ROS) free radicals in tissues and cells, leading to functional disorders, lesions and even death of multiple organs of the body. Therefore, ROS scavengers can protect against ionizing radiation damage.

[0003] Nanomaterial-based drug delivery systems have become a good alternative to traditional methods, which can reduce side effects and improve efficacy. Metal-organic frameworks (MOFs) have many potential advantages over other materials, such as diverse structures, high specific surface area, large pore sizes for drug loading, and inherent biodegradability.

[0004] In the entire field of radiation injury protection, only amifostine is used as a small molecule radiation protection drug. Although it has good efficacy, it has large side effects, a short half-life, and needs to be injected, which limits its application. At present, there are no approved oral radiation protection drugs, nor are there any approved low-dose radiation protection drugs. Radiation protection drugs belong to a semi-blank market, so it is of great significance to study protective drugs for ionizing radiation damage. Summary of the invention

[0005] The purpose of the embodiments of the present application is to provide a drug-carrying ligand, a chromium-based metal organic framework material and a preparation method thereof, aiming to solve the problems that existing radiation damage protection drugs have large side effects, short half-life, and need to be injected.

[0006] The embodiment of the present application is implemented as follows: a method for preparing a drug-carrying ligand comprises: Cysteine ​​methyl ester, HATU, 2,5-di(methoxyformyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12 h, and then washed and purified to obtain dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate; 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalic acid dimethyl ester was dissolved in THF / MeOH, and 3 mL of sodium hydroxide aqueous solution was added. The mixture was stirred at room temperature for 18 h, and then acidified and treated with concentrated acid under reduced pressure to obtain a drug-carrying ligand.

[0007] The present application also provides a method for preparing a chromium-based metal organic framework material, comprising: Dissolving chromium trichloride, terephthalic acid and the above-mentioned drug-carrying ligand in dimethylformamide for reaction for 1-48 hours at a reaction temperature of 80-250° C., washing and drying to obtain a chromium-based metal organic framework material; The molar ratio of the chromium trichloride, terephthalic acid and the drug-carrying ligand is 20:20:(1-20).

[0008] The embodiment of the present application also provides a chromium-based metal organic framework material, and the chromium-based metal organic framework material is prepared by the above-mentioned method for preparing the chromium-based metal organic framework material.

[0009] The embodiments of the present application also provide an application of the above-mentioned chromium-based metal organic framework material in protection against ionizing radiation damage.

[0010] An embodiment of the present application also provides a radiation protection drug, which includes the above-mentioned chromium-based metal-organic framework material, and one or more pharmaceutically acceptable carriers, adjuvants, auxiliary agents or diluents.

[0011] In the embodiment of the present application, cysteine ​​methyl ester, HATU, 2,5-di(methoxyformyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12 hours, washed and purified; 3 mL of sodium hydroxide aqueous solution was added to the obtained 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate in THF / MeOH, stirred at room temperature for 18 hours, acidified and treated with reduced pressure concentrated acid to obtain a drug-carrying ligand. The present application uses a drug-carrying ligand with a cysteine ​​active group to prepare metal organic framework nanoparticles, breaking through traditional physical adsorption or surface modification, and at the same time, a functional group with radiation protection ability is bonded to the ligand through ligand doping technology to achieve drug delivery. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1A schematic diagram of the synthesis of the chromium-based metal organic framework material provided in the embodiments of the present application; Figure 2 SEM image of the chromium-based metal organic framework material MOF-L provided in the examples of the present application (Example E); Figure 3 SEM image of the chromium-based metal organic framework material MOF-L provided in the examples of the present application (Example H); Figure 4 This is the in vitro radiation protection result of the chromium-based metal organic framework material MOF-L provided in the examples of the present application. DETAILED DESCRIPTION

[0013] In order to make the purpose, technical solution and advantages of the present application more clearly understood, the present application is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0014] The present application provides a method for preparing a drug-carrying ligand, comprising: Cysteine ​​methyl ester, HATU, 2,5-di(methoxyformyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12 h, and then washed and purified to obtain dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate; 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalic acid dimethyl ester was dissolved in THF / MeOH, and 3 mL of sodium hydroxide aqueous solution was added. The mixture was stirred at room temperature for 18 h, and then acidified and treated with concentrated acid under reduced pressure to obtain a drug-carrying ligand.

[0015] More specifically, the synthesis route and structural formula of the drug-carrying ligand are as follows: Cysteine ​​methyl ester (370 mg, 2.74 mmol), HATU (1.26 g, 3.32 mmol), 2,5-di(methoxyformyl)benzoic acid (625 mg, 2.63 mmol) and diisopropylethylamine (1.1 g, 8.3 mol) were stirred in DMF (10 ml) at room temperature for 12 h. The mixture was then diluted with water (10 ml) and ethyl acetate (30 ml). The organic layer was washed with saturated sodium bicarbonate and brine. The organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (methanol: dichloromethane = 1:30) to obtain a yellow solid (420 mg), dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate.

[0016] 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalic acid dimethyl ester (1000 mg, 2.8 mmol) was added to THF / MeOH (4 mL / 4 mL) with 3 ml of sodium hydroxide aqueous solution (30%) and stirred at room temperature for 18 h. The mixture was then concentrated under reduced pressure. Diluted with water, acidified to pH = 3 with concentrated hydrochloric acid, and then extracted with ethyl acetate. The organic phases were combined, dried over sodium sulfate, and concentrated under reduced pressure to obtain 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid.

[0017]

[0018] The present application also provides a method for preparing a chromium-based metal organic framework material, comprising the following steps: Chromium trichloride, terephthalic acid and the drug-carrying ligand prepared above are dissolved in dimethylformamide for reaction for 1-48 hours at a reaction temperature of 80-250° C. After washing and drying, a chromium-based metal organic framework material is obtained.

[0019] In the embodiment of the present application, the structure of the prepared chromium-based metal organic framework material is Cr3Cl(H2O)2O[(O2C)-C6H4-(CO2)]3(C 12 H 11 NO7S) n , the benzoic acid group of the drug-carrying ligand is coordinated with the chromium in the metal organic framework.

[0020] The drug-carrying ligand is a benzoic acid structure compound containing a small molecule ROS scavenger structure, which is 2-[[(1-carboxyl-2-mercaptoethyl)carbonyl imide] formyl] terephthalic acid.

[0021] In an embodiment of the present application, the molar ratio of the chromium trichloride, terephthalic acid, and drug-carrying ligand is 20:20:(1-20), for example, the molar ratio may be 20:20:1, 20:20:2, 20:20:3, 20:20:4, 20:20:5, 20:20:6, 20:20:7, 20:20:8, 20:20:9, 20:20:10, 20:20:11, 20:20:12, 20:20:15, 20:20:16, 20:20:17, 20:20:18, 20:20:19, 20:20:20, etc. Preferably, the molar ratio of the chromium trichloride, terephthalic acid, and drug-carrying ligand is 2:2:1. It is worth noting that the molar ratio of the chromium trichloride, terephthalic acid and the drug-carrying ligand directly affects the crystal form or drug loading of the obtained material. For example, if the molar ratio of 2-[[(1-carboxy-2-mercaptoethyl)carbonylimide]formyl]terephthalic acid is too large, the crystal form of the obtained material is poor; if the molar ratio of 2-(2,2-dimethylthiazolidine-3-carbonyl)terephthalic acid is too small, the drug loading of the obtained material is small.

[0022] In an embodiment of the present application, chromium trichloride, terephthalic acid and a drug-carrying ligand are added to dimethylformamide, and after ultrasonic dissolution, a heating and stirring reaction is performed to form a brown solid. After washing with distilled water and ethanol, the metal organic framework nanoparticles are obtained by vacuum drying. Among them, the dissolution temperature can be 20-100°C, and after sufficient dissolution, the reaction is carried out at a reaction temperature of 80-250°C for 1-48h. Among them, when the reaction temperature is lower than 80°C, the system cannot react. When the reaction temperature is higher than 250°C, the crystal form of the chromium-based metal organic framework material obtained is poor. Preferably, the reaction temperature is 170°C and the reaction time is 15h.

[0023] In a specific embodiment of the present application, the organic solvent is a suitable solvent commonly used in organic reactions, for example, including but not limited to aliphatic and aromatic, optional hydrocarbons or halogenated hydrocarbons (such as pentane, hexane, heptane, cyclohexane, petroleum ether, gasoline, volatile oil, benzene, toluene, xylene, dichloromethane, dichloroethane, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene), aliphatic and aromatic, optional alcohols (such as methanol, ethanol, propanol, isopropanol, tert-butanol, ethylene glycol, etc.), ethers (such as diethyl ether and dibutyl ether, ethylene glycol dimethyl ether and diethylene glycol dimethyl ether, tetrahydrofuran and dioxane, etc.), esters (such as methyl acetate or ethyl acetate, etc.), nitriles (such as acetonitrile or propionitrile, etc.), ketones (such as acetone, butanone, etc.), amides (such as dimethylformamide, dimethylacetamide and N-methylpyrrolidone, etc.), and dimethyl sulfoxide, tetramethylene sulfone, etc. An embodiment of the present application also provides a radiation protection drug, which includes the above-mentioned chromium-based metal-organic framework material, and one or more pharmaceutically acceptable carriers, adjuvants, auxiliary agents or diluents.

[0024] In a specific embodiment of the present application, the dosage form of the pharmaceutical composition includes but is not limited to injection, emulsion, microemulsion, submicroemulsion, nanoparticles, tablets, capsules, pills, inhalants, lozenges, gels, powders, suppositories, suspensoids, creams, jellies or sprays.

[0025] In a specific embodiment of the present application, the pharmaceutical composition can be administered in ways including, but not limited to, subcutaneous injection, intramuscular injection, intravenous injection, oral administration, rectal administration, vaginal administration, nasal administration, transdermal administration, subconjunctival administration, intraocular administration, orbital administration, retro-ocular administration, retinal administration, choroidal administration or intrathecal injection.

[0026] The embodiments of the present application also provide a use of the above-mentioned chromium-based metal-organic framework material or radiation protection drug in the preparation of a drug for treating and / or preventing ionizing radiation damage.

[0027] Examples of certain embodiments of the present application are given below, which are not intended to limit the scope of the present application.

[0028] In addition, it should be noted that the numerical values ​​given in the following examples are as accurate as possible, but those skilled in the art understand that due to unavoidable measurement errors and experimental operation problems, each number should be understood as an approximate number rather than an absolutely accurate value.

[0029] Preparation of drug-loaded ligands: Cysteine ​​methyl ester (370 mg, 2.74 mmol), HATU (1.26 g, 3.32 mmol), 2,5-di(methoxyformyl)benzoic acid (625 mg, 2.63 mmol) and diisopropylethylamine (1.1 g, 8.3 mol) were stirred in DMF (10 ml) at room temperature for 12 h. The mixture was then diluted with water (10 ml) and ethyl acetate (30 ml). The organic layer was washed with saturated sodium bicarbonate and brine. The organic phase was dried and concentrated to obtain a crude product, which was purified by column chromatography (methanol: dichloromethane = 1:30) to obtain a yellow solid (420 mg), dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate, with a yield of 43%.

[0030] 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalic acid dimethyl ester (1000 mg, 2.8 mmol) was added to THF / MeOH (4 mL / 4 mL) with 3 ml of sodium hydroxide aqueous solution (30%) and stirred at room temperature for 18 h. The mixture was then concentrated under reduced pressure. Diluted with water, acidified to pH = 3 with concentrated hydrochloric acid, and then extracted with ethyl acetate. The organic phases were combined, dried over sodium sulfate, and concentrated under reduced pressure to obtain 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid (drug-carrying ligand), 610 mg of brown solid, with a yield of 69%. 1 H NMR (400MHz, CDCl3) δ 10.50 (bs, 3H), 8.48 (d, 1H), 8.35 (m, 2H), 8.00 (s, 1H), 4.60 (t, 1H), 3.50 (d, 2H). ESI-MS: m / z= 336 (M+Na)+; HRMS (ESI): m / z [M+H]+ calcd for C 12 H 11 NO7S: 313.0256, found: 313.0261. Example 1 Preparation of chromium-based metal organic framework materials like Figure 1 As shown, the chromium-based metal organic framework material provided in this embodiment is Cr3Cl(H2O)2O[(O2C)-C6H4-(CO2)]3(C 12 H 11 NO7S) n Abbreviated as MOF-L (n=0.05-1).

[0031] The preparation method of the chromium-based metal organic framework material comprises adding chromium trichloride hexahydrate, 2-[[(1-carboxyl-2-mercaptoethyl)carbonyl imide]formyl]terephthalic acid, and terephthalic acid into dimethylformamide to dissolve. The molar ratio of chromium trichloride:2-[[(1-carboxyl-2-mercaptoethyl)carbonyl imide]formyl]terephthalic acid):terephthalic acid in this embodiment is 20:1~20:20. The molar ratio of chromium trichloride:2-[[(1-carboxyl-2-mercaptoethyl)carbonyl imide]formyl]terephthalic acid:terephthalic acid ligand in the specific example is shown in Table 1 below.

[0032] Table 1 Molar ratio of chromium trichloride: 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid:terephthalic acid in each example Examples Chromium trichloride (mol) Terephthalic acid (mol) 2-[[(1-Carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid (mol) A 1 1 0.05 B 1 1 0.1 C 1 1 0.2 D 1 1 0.4 E 1 1 0.5 F 1 1 0.8 G 1 1 1 H 1 1 2 like Figure 2 , Figure 3 In the SEM image shown, Figure 2 :MOF-L (Example E), Figure 3 :MOF-L (Example H). The molar ratio of 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid shown in Example H is too large, and the crystal form shown in the SEM image is worse than that in Example E; but if the molar ratio of 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid is too small, the drug loading is small. Therefore, the present application prefers that the molar ratio of chromium trichloride:2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid) is 20:10:20.

[0033] The following is a detailed description of the preparation method of chromium-based metal organic framework material (denoted as MOF-L) using Example E as an example.

[0034] In a 30 ml autoclave, add CrCl3·6H2O (0.27 g), 2-[[(1-carboxy-2-mercaptoethyl)carbonyl imide] formyl] terephthalic acid (0.16 g), terephthalic acid (0.17 g) and dimethylformamide (30 ml), and then ultrasonically dissolve the reaction system for 15 minutes. The reaction solution is heated to 170°C, and the reactor is stirred in a magnetic stirrer for 15 hours. The reaction solution is cooled to room temperature, centrifuged at 1500 rpm for 30 minutes, and the supernatant is discarded. Add 15 ml of distilled water to the obtained solid, stir and centrifuge at 1500 rpm for 30 minutes, and discard the supernatant. Add 10 ml of ethanol to the obtained solid, stir and centrifuge at 3000 rpm for 30 minutes, and discard the supernatant. Add dimethylformamide (20 ml) to the obtained solid, stir at room temperature for 48 hours, centrifuge at 1500 rpm for 30 minutes, and discard the supernatant. The obtained solid was added with ethanol (10 ml) and stirred, then centrifuged at 800 rpm for 30 minutes, the supernatant was discarded, and vacuum dried to obtain a brown solid (0.25 g), which is MOF-L. The SEM characterization of MOF-L is shown in Figure 2 shown.

[0035] Example 3 In vitro cytotoxicity assay of drug compounds Using amifostine as a positive control, the cytotoxicity of the prepared MOF nanoparticles in HFL-1 was detected and its IC50 value was calculated. The experimental results are shown in Table 2.

[0036] Table 2 Cytotoxicity test of four MOF-loaded drugs in HFL-1 Compound No. IC50 value (μM) Drug 1 MOF-L >100 Comparison Amifostine >100 As can be seen from Table 2, the cytotoxicity of MOF-loaded drugs is relatively low, similar to amifostine, and no cytotoxicity was observed at a concentration of 100 μM.

[0037] Example 4 In vivo radiation protection of pharmaceutical compounds An in vivo mouse radiation model was established to study the in vivo radiation protection effect of MOF-L. The whole-body irradiation with 137Cs method was used to establish a mouse radiation model to observe the effect of MOF-L on the survival rate of mice. C57BL / 6 mice were divided into groups according to the random block method of body weight. The irradiation + MOF-L injection group (denoted as a) received 7.2 Gy whole-body irradiation, and MOF-L was intraperitoneally injected 1 h before irradiation; the irradiation + MOF-L gavage group (denoted as b) received 7.2 Gy whole-body irradiation, and MOF-L was gavaged 1 h before irradiation; the irradiation group (7.2 Gy) (denoted as c) received 7.2 Gy whole-body irradiation, and normal saline was gavaged 1 h before irradiation; during irradiation, the mice received a single 137Cs source whole-body irradiation (TBI) with an irradiation dose of 7.2 Gy and a dose rate of 0.99 Gy / min. The death and body weight of the mice were recorded daily, and the 30-day survival rate of the mice was calculated. The results are shown in the figure. Figure 4 As shown. Figure 4 It can be seen that the drug compound MOF-L has an in vitro radiation protection effect both by oral gavage and intraperitoneal injection.

[0038] The above-mentioned embodiments only express several implementation methods of the present application, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the present application. It should be pointed out that, for a person of ordinary skill in the art, several variations and improvements can be made without departing from the concept of the present application, and these all belong to the protection scope of the present application. Therefore, the protection scope of the present application shall be subject to the attached claims.

[0039] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method for preparing a drug-carrying ligand, characterized in that: include: Cysteine ​​methyl ester, HATU, 2,5-di(methoxyformyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12 h, and then washed and purified to obtain dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate; 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalic acid dimethyl ester was dissolved in THF / MeOH, and 3 mL of sodium hydroxide aqueous solution was added. The mixture was stirred at room temperature for 18 h, and then acidified and treated with concentrated acid under reduced pressure to obtain a drug-carrying ligand.

2. A drug-carrying ligand, characterized in that: The drug-carrying ligand is prepared by the preparation method of the drug-carrying ligand according to claim 1.

3. A method for preparing a chromium-based metal organic framework material, characterized in that: include: Dissolving chromium trichloride, terephthalic acid, and the drug-carrying ligand according to claim 2 in dimethylformamide for reaction for 1-48 hours at a reaction temperature of 80-250° C., washing, and drying to obtain a chromium-based metal organic framework material; The molar ratio of the chromium trichloride, terephthalic acid and the drug-carrying ligand is 20:20:(1-20).

4. The method for preparing a chromium-based metal organic framework material according to claim 3, characterized in that: The molar ratio of the chromium trichloride, terephthalic acid and the drug-carrying ligand is 2:2:

1.

5. The method for preparing a chromium-based metal organic framework material according to claim 3, characterized in that: The reaction temperature was 170°C and the reaction time was 15 h.

6. A chromium-based metal organic framework material, characterized in that: The chromium-based metal organic framework material is prepared by the preparation method of the chromium-based metal organic framework material according to any one of claims 3-5.

7. A radiation protection drug, characterized in that: The radiation protection drug comprises the chromium-based metal organic framework material according to claim 6, and one or more pharmaceutically acceptable carriers, adjuvants, auxiliary agents or diluents.

8. Use of the chromium-based metal organic framework material according to claim 6 or the radiation protection drug according to claim 7 in the preparation of a drug for treating and / or preventing ionizing radiation damage.

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