Drug-loaded ligand, chromium-based metal-organic framework material and preparation method and application thereof

Abstract

The application is suitable for 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, the preparation method of the drug-loaded ligand comprises the following steps: stirring cysteine methyl ester, HATU, 2,5-di(methoxycarbonyl)benzoic acid and diisopropylethylamine in DMF at room temperature for 12 h, and performing washing and purification treatment; the obtained 2-((3-mercapto-1-methoxy-1-oxoprop-2-yl)carbamoyl) terephthalic acid dimethyl ester is added with 3 mL of a sodium hydroxide aqueous solution in THF / MeOH, stirred at room temperature for 18 h, and subjected to acidification and concentrated acid treatment under reduced pressure to obtain the drug-loaded ligand. The application breaks through the traditional physical adsorption or surface modification by adopting the drug-loaded ligand with a cysteine active group to prepare metal organic framework nanoparticles, and simultaneously realizes drug administration by bonding a functional group with radiation protection capability to the ligand through a ligand doping technology.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of materials, and particularly relates to a drug-loaded ligand, a chromium-based metal organic framework material and a preparation method and application thereof. BACKGROUND

[0002] Drug release systems based on nanomaterials have become a good choice to replace traditional methods, which can reduce side effects and improve efficacy. Metal organic frameworks (MOFs) have many potential advantages compared with other materials, such as structural diversity, high specific surface area, large pore size for drug loading, and inherent biodegradability. In 2005, Ferey's group first reported that a series of MOF metal organic framework materials were used as potential drug carriers, and since then, research on MOFs as drug carriers has been emerging. With the continuous development of world nuclear application technology, especially the emergence of nuclear power plants, radiation processing technology, and the increasing trend of radiation accidents. On the other hand, the number of cancer patients receiving radiotherapy is also increasing year by year. Therefore, the research on radiation protection drugs is not only the demand of national nuclear safety, but also closely related to people's daily life, and is a major medical problem that should be paid enough attention to. Ionizing radiation can produce reactive oxygen species (ROS) free radicals in tissues and cells, leading to functional disorders, pathological changes, and even death of the body, so ROS scavengers can prevent ionizing radiation damage.

[0003] Drug release systems based on nanomaterials have become a good choice to replace traditional methods, which can reduce side effects and improve efficacy. Metal organic frameworks (MOFs) have many potential advantages compared with other materials, such as structural diversity, high specific surface area, large pore size for drug loading, and inherent biodegradability.

[0004] In the field of radiation damage protection, only amifostine is used as a small molecule radiation protection drug. Although it has good efficacy, it has large side effects, short half-life, and needs to be injected, which limits its application. At present, there is no approved oral radiation protection drug and no approved low-dose radiation protection drug. Radiation protection drugs belong to a semi-blank market, so it is of great significance to study the protection drugs for ionizing radiation damage. SUMMARY

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

[0006] The embodiments of the present application are implemented in this way, a preparation method of a drug-loaded ligand, comprising:

[0007] Cysteine methyl ester, HATU, 2,5-di(methoxycarbonyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12h, and after washing and purification, dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate was obtained;

[0008] Dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate was stirred in THF / MeOH, 3mL of aqueous sodium hydroxide was added, and stirred at room temperature for 18h, and after acidification and concentrated acid treatment under reduced pressure, the drug-loaded ligand was obtained.

[0009] The application also provides a preparation method of a chromium-based metal organic framework material, comprising:

[0010] The chromium trichloride, terephthalic acid and the drug-loaded ligand are dissolved in dimethylformamide for reaction for 1-48h, the reaction temperature is 80-250 DEG C, and after washing and drying treatment, the chromium-based metal organic framework material is obtained.

[0011] The molar ratio of the chromium trichloride, terephthalic acid and the drug-loaded ligand is 20:20:(1-20).

[0012] The application also provides a chromium-based metal organic framework material prepared by the above preparation method of the chromium-based metal organic framework material.

[0013] The application also provides an application of the above chromium-based metal organic framework material in ionizing radiation damage protection.

[0014] The application also provides a radiation protection drug, which comprises the above chromium-based metal organic framework material, and one or more pharmaceutically acceptable medium carriers, adjuvants, auxiliaries or diluents.

[0015] The application comprises the following steps: cysteine methyl ester, HATU, 2,5-di(methoxycarbonyl)benzoic acid and diisopropylethylamine are stirred in DMF at room temperature for 12h, and after washing and purification, dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate is obtained; dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate is stirred in THF / MeOH, 3mL of aqueous sodium hydroxide is added, and stirred at room temperature for 18h, and after acidification and concentrated acid treatment under reduced pressure, the drug-loaded ligand is obtained. The application breaks through the traditional physical adsorption or surface modification by adopting the drug-loaded ligand with a cysteine active group to prepare the metal organic framework nanoparticles, and realizes drug administration by bonding the functional groups with radiation protection capacity to the ligand through the ligand doping technology. BRIEF DESCRIPTION OF DRAWINGS

[0016] Figure 1 A synthesis diagram of a chromium-based metal organic framework material provided for an embodiment of the present application is shown in the following figure:

[0017] Figure 2 A SEM image of the chromium-based metal organic framework material MOF-L provided for an embodiment of the present application (Example E) is shown in the following figure:

[0018] Figure 3 A SEM image of the chromium-based metal organic framework material MOF-L provided for an embodiment of the present application (Example H) is shown in the following figure:

[0019] Figure 4 An in vitro radiation protection result of the chromium-based metal organic framework material MOF-L provided for an embodiment of the present application is shown in the following table: DETAILED DESCRIPTION

[0020] In order to make the purpose, technical scheme and advantages of the present application more clear, the present application is further described in detail below in combination with the 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.

[0021] The present application provides a preparation method of a drug-loaded ligand, comprising:

[0022] Cysteine methyl ester, HATU, 2,5-di(methoxycarbonyl)benzoic acid and diisopropylethylamine were stirred in DMF at room temperature for 12 h, and then washed and purified to obtain 2-((3-mercapto-1-methoxy-1-oxoprop-2-yl)carbamoyl) terephthalic acid dimethyl ester.

[0023] 2-((3-mercapto-1-methoxy-1-oxoprop-2-yl)carbamoyl) terephthalic acid dimethyl ester was added with 3 mL of sodium hydroxide aqueous solution in THF / MeOH, and stirred at room temperature for 18 h, and then acidified and concentrated under reduced pressure to obtain the drug-loaded ligand.

[0024] More specifically, the synthesis route and structural formula of the drug-loaded ligand are as follows:

[0025] Cysteine methyl ester (370 mg, 2.74 mmol), HATU (1.26 g, 3.32 mmol), 2,5- bis(methoxycarbonyl)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 the addition of water (10 mL) and ethyl acetate (30 mL). The organic layer was washed with saturated sodium bicarbonate, brine. The organic phase was dried and concentrated to give a crude product, which was purified by column chromatography (methanol: dichloromethane = 1:30) to give a yellow solid (420 mg), dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate.

[0026] Dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate (1000 mg, 2.8 mmol) was dissolved in THF / MeOH (4 mL / 4 mL), 3 ml of aqueous sodium hydroxide solution (30%) was added, and stirred at room temperature for 18 h. Then the mixture was concentrated under reduced pressure. Diluted with water, acidified to pH = 3 with concentrated hydrochloric acid, and then extracted with ethyl acetate. The organic phase was combined, dried over sodium sulfate, and concentrated under reduced pressure to give 2-[[(1-carboxy-2-mercaptoethyl)carbonylimino]formyl]terephthalic acid.

[0027]

[0028] The application also provides a preparation method of a chromium-based metal organic framework material, comprising the following steps:

[0029] The chromium trichloride, terephthalic acid and the prepared drug-loaded ligand are dissolved in dimethylformamide for reaction for 1-48 h, the reaction temperature is 80-250°C, and the chromium-based metal organic framework material is obtained through washing and drying treatment.

[0030] In the embodiment of the 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-loaded ligand is coordinated with the chromium in the metal organic framework.

[0031] The drug-loaded ligand is a benzoic acid compound containing a small molecule ROS scavenger structure, which is 2-[[(1-carboxy-2-mercaptoethyl)carbonylimino]formyl]terephthalic acid.

[0032] In the embodiments of the present application, the molar ratio of the chromium trichloride, terephthalic acid and drug-loaded ligand is 20:20:(1-20), for example, the molar ratio can 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-loaded ligand is 2:2:1. It is worth noting that the molar ratio of the chromium trichloride, terephthalic acid and drug-loaded ligand directly affects the crystal form or drug loading amount of the obtained material, for example, if the molar ratio of 2-[[(1-carboxy-2-mercaptoethyl) carbonylimino] 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 amount of the obtained material is small.

[0033] In the embodiments of the present application, the chromium trichloride, terephthalic acid and drug-loaded ligand are added to dimethylformamide, and after ultrasonic dissolution, 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. The dissolution temperature can be 20-100°C, and after sufficient dissolution, the reaction is performed at a reaction temperature of 80-250°C for 1-48h. When the reaction temperature is lower than 80°C, the system cannot react, and 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.

[0034] In one 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, optionally hydrocarbon or halogenated hydrocarbon (for example, pentane, hexane, heptane, cyclohexane, petroleum ether, gasoline, volatile oil, benzene, toluene, xylene, dichloromethane, dichloroethane, chloroform, carbon tetrachloride, chlorobenzene and o-dichlorobenzene), aliphatic and aromatic, optionally alcohol (for example, methanol, ethanol, propanol, isopropanol, tert-butyl alcohol, ethylene glycol, etc.), ether (for example, diethyl ether and dibutyl ether, ethylene glycol dimethyl ether and diethylene glycol dimethyl ether, tetrahydrofuran and dioxane, etc.), ester (for example, methyl acetate or ethyl acetate, etc.), nitrile (for example, acetonitrile or propionitrile, etc.), ketone (for example, acetone, butanone, etc.), amide (for example, dimethylformamide, dimethylacetamide and N-methylpyrrolidone, etc.), and dimethyl sulfoxide, tetramethylene sulfone, etc.

[0035] The embodiment of the present application also provides a radiation protection drug, which comprises the above-mentioned chromium-based metal organic framework material and one or more pharmaceutically acceptable carriers, adjuvants, assistants or diluents.

[0036] In one specific embodiment of the present application, the dosage form of the pharmaceutical composition includes, but is not limited to, an injection, an emulsion, a microemulsion, a submicroemulsion, a nanoparticle, a tablet, a capsule, a pill, an inhalant, a lozenge, a gel, a powder, a suppository, a suspoemulsion, a cream, a jelly or a spray.

[0037] In one specific embodiment of the present application, the administration mode of the pharmaceutical composition can include, but is not limited to, subcutaneous injection, intramuscular injection, intravenous injection, oral administration, rectal administration, vaginal administration, nasal administration, transdermal administration, subconjunctival administration, intraocular administration, intraorbital administration, retrobulbar administration, retinal administration, choroidal administration or intrathecal injection.

[0038] The embodiment of the present application also provides use of the above-mentioned chromium-based metal organic framework material or the radiation protection drug in the preparation of a drug for treating and / or preventing ionizing radiation damage.

[0039] The following gives examples of some embodiments of the present application, which are not intended to limit the scope of the present application.

[0040] 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 number.

[0041] Preparation of drug-loaded ligand:

[0042] Cysteine methyl ester (370 mg, 2.74 mmol), HATU (1.26 g, 3.32 mmol), 2,5-di(methoxycarbonyl)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 the addition of water (10 mL) and ethyl acetate (30 mL). The organic layer was washed with saturated sodium bicarbonate, brine. The organic phase was dried and concentrated to give a crude product, which was purified by column chromatography (methanol: dichloromethane = 1:30) to obtain a yellow solid (420 mg), 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)dimethyl terephthalate, with a yield of 43%.

[0043] To dimethyl 2-((3-mercapto-1-methoxy-1-oxopropan-2-yl)carbamoyl)terephthalate (1000 mg, 2.8 mmol) in THF / MeOH (4 mL / 4 mL), 3 mL of 30% aqueous sodium hydroxide solution was added and stirred at room temperature for 18 hours. The mixture was then concentrated under reduced pressure. The mixture was diluted with water, acidified to pH 3 with concentrated hydrochloric acid, and extracted with ethyl acetate. The combined organic phases were dried over sodium sulfate and concentrated under reduced pressure to yield 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid (drug-carrying ligand) as a brown solid (610 mg, 69% yield). 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.

[0044] Example 1 Preparation of chromium-based metal-organic framework materials

[0045] 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).

[0046] The preparation method of a chromium-based metal-organic framework material comprises dissolving chromium trichloride hexahydrate, 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid, and terephthalic acid in dimethylformamide. In this embodiment, the molar ratio of chromium trichloride:2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid:terephthalic acid is 20:1 to 20:20. The molar ratios of chromium trichloride:2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid:terephthalic acid ligands in a specific example are shown in Table 1 below.

[0047] Table 1 Molar ratio of chromium trichloride: 2-[[(1-carboxy-2-mercaptoethyl)carbonylimido]formyl]terephthalic acid: terephthalic acid in each example

[0048] Example chromium trichloride (mol) terephthalic acid (mol) 2-[[(1-carboxy-2-mercaptoethyl)carbonylimino]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

[0049] As shown in the SEM images of Figure 2 , Figure 3 , Figure 2 : MOF-L (Example E), Figure 3 : MOF-L (Example H). The molar ratio of 2-[[(1-carboxy-2-mercaptoethyl) carbonylimino] formyl] terephthalic acid in Example H is too large, and the SEM image shows a worse crystal form than Example E; but if the molar ratio of 2-[[(1-carboxy-2-mercaptoethyl) carbonylimino] formyl] terephthalic acid is too small, the drug loading amount is small. Therefore, the molar ratio of CrCl3: 2-[[(1-carboxy-2-mercaptoethyl) carbonylimino] formyl] terephthalic acid): terephthalic acid in the present application is preferably 20:10:20.

[0050] The preparation method of the chromium-based metal organic framework material (denoted as MOF-L) will be described in detail below taking Example E as an example.

[0051] In a 30-mL high-pressure reaction kettle, CrCl3·6H2O (0.27 g), 2-[[(1-carboxy-2-mercaptoethyl) carbonylimino] formyl] terephthalic acid (0.16 g), terephthalic acid (0.17 g), and dimethylformamide (30 mL) were added, and then the reaction system was ultrasonically dissolved for 15 minutes. The reaction liquid was heated to 170°C, and the reaction kettle was stirred in a magnetic stirrer for 15 hours. The reaction liquid was cooled to room temperature, centrifuged at 1500 rpm for 30 minutes, and the supernatant was discarded. The obtained solid was added with distilled water (15 mL), stirred, and then centrifuged at 1500 rpm for 30 minutes, and the supernatant was discarded. The obtained solid was added with ethanol (10 mL), stirred, and then centrifuged at 3000 rpm for 30 minutes, and the supernatant was discarded. The obtained solid was added with dimethylformamide (20 mL), stirred at room temperature for 48 hours, and then centrifuged at 1500 rpm for 30 minutes, and the supernatant was discarded. The obtained solid was added with ethanol (10 mL), stirred, and then centrifuged at 800 rpm for 30 minutes, and the supernatant was discarded, and vacuum drying was performed to obtain a brown solid (0.25 g), which was MOF-L. The SEM characterization of MOF-L is shown in Figure 2 .

[0052] Example 3 In vitro cytotoxicity determination of drug compounds

[0053] With amifostine as a positive control, the cytotoxicity of the prepared MOF nanoparticles in HFL-1 was detected, and the IC50 value was calculated, and the experimental results are shown in Table 2.

[0054] Table 2 Cytotoxicity test of four MOF-loaded drugs in HFL-1

[0055] Compound No. IC50 value (μM) Drug 1 MOF-L >100 Control Amifostine >100

[0056] From Table 2, it can be seen that the cytotoxicity of MOF loaded drugs is small, similar to amifostine, and no cytotoxicity is observed at a concentration of 100 μM.

[0057] Example 4 In vivo radiation protection effect of drug compounds

[0058] An in vivo mouse radiation model was established to study the in vivo radiation protection effect of MOF-L. A mouse radiation model was established using whole-body irradiation with 137Cs, and the effect of MOF-L on the survival rate of mice was observed. C57BL / 6 mice were randomly grouped according to body weight, and the following groups were established: irradiation + MOF-L injection group (denoted as a): 7.2 Gy whole-body irradiation was received, and MOF-L was injected intraperitoneally 1 h before irradiation; irradiation + MOF-L gavage group (denoted as b): 7.2 Gy whole-body irradiation was received, and MOF-L was administered by gavage 1 h before irradiation; irradiation group (7.2 Gy) (denoted as c): 7.2 Gy whole-body irradiation was received, and normal saline was administered by gavage 1 h before irradiation; the mice received whole-body irradiation (TBI) from a 137Cs source at a 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, and the results are shown in Figure 4 From the results shown in Figure 4 , it can be seen that the drug compound MOF-L administered by gavage and intraperitoneal injection both have in vitro radiation protection effects.

[0059] The above-described embodiments only express several embodiments of the present application, and the description is relatively specific and detailed, but it should not be understood as limiting the scope of the patent of the present application. It should be noted that, for ordinary skilled persons in the art, without departing from the concept of the present application, several modifications and improvements can be made, which are all within the protection scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

[0060] The above-described only preferred embodiments of the present application, and does not limit the present application, any modification, equivalent replacement and improvement 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: Methyl cysteine, 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, acidified, and treated with concentrated acid under reduced pressure to obtain a drug-loaded 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 and reacting 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 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 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 and adjuvants.

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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