CN114907573B - Metal organic framework material and application thereof in treatment of fungal keratitis - Google Patents
Metal organic framework material and application thereof in treatment of fungal keratitis Download PDFInfo
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Abstract
The application discloses a preparation method of a metal organic framework material, which comprises the steps of firstly preparing meso-tetra (4-carboxyphenyl) porphyrin and ZrOCl 2 ·8H 2 Dissolving O, benzoic acid and polyene antibiotics in dimethylformamide for mixed reaction, centrifuging and taking solid to obtain a crude product; and then washing the crude product to obtain the metal organic framework material. The application also discloses a metal organic framework material which is worthy of the preparation method and application thereof in treating fungal keratitis. The preparation method is simple, the coordination is realized by a one-pot method, the polyene antifungal agent is arranged in the organic metal frame in a coordination form, the medicine carrying rate is improved, the polyene antifungal agent can be tightly combined with the cornea mucin, so that the detention time is prolonged, the permeability of the polyene antifungal agent in the cornea is increased, the solubility and the dispersibility of the antifungal agent are improved, the synergistic sterilization of the chemical treatment and the photodynamic treatment is realized, and the efficient sterilization is realized.
Description
Technical Field
The application relates to the field of beauty equipment, in particular to a metal organic framework material and application thereof in treating fungal keratitis.
Background
The cornea is positioned at the front part of the eyeball and is in direct contact with the outside, so that the cornea is easily damaged to damage the corneal epithelium, thereby causing infectious keratitis. Infectious keratopathy is one of the major causes of blindness, including fungi, bacteria, viruses, etc. Among the infectious keratopathy, mycotic keratitis is the most serious destructive disease due to long disease course, great harm, difficult cure and serious complications thereof, and is the second most common blinding cause in developing countries. The incidence rate of the fungal keratitis is high (about 100 ten thousand cases of cornea infection are newly increased every year), and the blindness rate is high, which accounts for 1-45% of the infectious keratitis. The incidence of fungal keratitis is higher in 21 developing countries than in developed countries. Compared to other infectious keratitis, the following differences exist: firstly, fungal pathogens are powerful themselves, almost always causing infection once they enter the corneal barrier; second, secondary damage caused by mycotoxins and enzymes often results in ulcers, perforations, and pus accumulation in the anterior chamber, even blindness; thirdly, the development time is two weeks to several months, so that strong immune response is often caused, and the formation of new blood vessels and cornea scars is often accompanied, so that the vision is seriously affected. In such severe cases, however, topical administration remains the most prominent therapeutic modality.
Antifungals are classified according to their molecular structure and mechanism of action, and are classified into polyenes (amphotericin B, natamycin), azoles (voriconazole, fluconazole, itraconazole), pyrimidines (5-fluorocytosine), allylamines (terbinafine), echinococcins (caspofungin) and heterocyclic benzofurans (griseofulvin). Among antifungal drugs for the treatment of fungal ocular infections (fungal keratitis, endophthalmitis, conjunctivitis, blepharitis), polyene antifungal drugs are the main therapeutic drugs due to their broad antifungal spectrum and powerful biological activity. Polyene antifungal agents have two characteristics, namely, the presence of multiple conjugated double bonds in the hydroxyl chromophore, which are necessary for their antibacterial activity and stability; and the second is a zwitterionic substance which contains a moldy amine group and a carboxylic acid group connected by ether bonds and imparts amphiprotic properties to the moldy amine group and the carboxylic acid group, and has an isoelectric point at pH of 5-7.
However, intraocular administration of polyene antifungal agents still presents numerous challenges. The different levels of cornea, sclera, retina, blood-retina, and blood aqueous barrier constitute anatomical barriers, while ocular blood flow, tear dilution, ocular enzymes, and transport proteins together constitute physiological barriers. These blood-ocular barriers consist of tight endothelial junctions, limiting the movement of high molecular weight antifungal drugs. Wherein the physiological barrier also prevents ocular administration of the drug, and the drainage of nasal tears results in considerable loss when the polyene is administered topically, the tear film reduces the antifungal residence time. On the other hand, the antifungal drug itself has physical and chemical limitations, and has problems of poor permeability, poor solubility and poor dispersibility. These problems result in a substantial decrease in the bioavailability of polyene antifungal drugs, less than 5% and even 2%. Therefore, the treatment of the fungal keratitis requires frequent use of antifungal medicines, and the antifungal medicines are used once every 1-2 hours in the first three and four days, and the treatment is reduced to 6-8 times per day in the next week until the treatment is completed. Thus, on the one hand, good patient compliance is required and on the other hand, there is also a great deal of drug toxicity from frequent long-term administration.
In response to the above problems, more and more ophthalmic administration strategies have been developed in recent years for treating fungal keratitis, and they fall into three general categories: firstly, the innovation of the administration modes, such as carrier implantation administration, ocular ion introduction, contact lens administration and microneedle administration, can improve certain bioavailability through verification, but the defects are obvious; the carrier implantation administration has certain requirements on the operation level of doctors, and patients have certain uncomfortable feeling while being invasive; the ocular ion introduction has certain requirements on the operation level of doctors, and has more limiting factors; the administration of the contact lens has certain requirements on the light transmittance and the water absorbability of the contact lens, and the administration of the contact lens has certain inconvenience for patients to wear when the drug loading rate is not high enough; microneedle administration has a certain heat degree in recent years, and minimally invasive operation has a certain attractive force, however, in order to truly improve bioavailability, the close relationship with the selected materials and types is realized, the problem that the drug loading capacity is low is difficult to solve, and the acceptance of patients on microneedle patches is low. Secondly, the innovation of the drug delivery carrier, such as liposome drug delivery, micelle drug delivery and the like, can improve the bioavailability while being convenient for drug delivery, has more researches in recent years, but has the disadvantage of not being negligible. The liposome drug loading can break through the cornea barrier, but the liposome is not stable enough and the drug loading rate is not high enough, so the drug amount entering the stroma layer is not high. The micelle can stably release the medicine and improve the slow release time, but the high molecular polymer has high price and simultaneously the bioavailability is not obviously improved. Thirdly, innovations of treatment methods, such as photodynamic, photothermal, cornea collagen crosslinking, mesenchymal stem cell injection and the like; the pure photodynamic and photothermal effects have certain therapeutic effects, but the good therapeutic effects are achieved at the cost of great damage to normal cells, and the cytotoxicity limits the application of the pure photodynamic and photothermal effects; cornea collagen crosslinking is mainly used as an auxiliary treatment, fungal keratitis is easy to ulcer perforation, cornea collagen crosslinking can effectively limit ulcer development, but can also only be used as an auxiliary treatment; mesenchymal stem cell injection has been the only concept in recent years, and its application effect has been questionable.
Disclosure of Invention
Therefore, the technical problem to be solved by the application is to solve the problem that the existing polyene medicine has poor effect of treating the fungal keratitis, thereby providing a metal organic framework material and application thereof in treating the fungal keratitis.
Therefore, the application adopts the following technical scheme:
the application provides a preparation method of a metal organic framework material, which comprises the following steps:
s1: meso-tetra (4-carboxyphenyl) porphyrin and ZrOCl 2 ·8H 2 Dissolving O, benzoic acid and polyene antibiotics in dimethylformamide for mixed reaction, centrifuging and taking solid to obtain a crude product;
s2: washing the crude product with dimethylformamide to obtain the metal organic framework material.
Preferably, the polyene antibiotic is natamycin.
Further, in step S1, the meso-tetra (4-carboxyphenyl) porphyrin, zrOCl 2 ·8H 2 O, benzoic acid and thatThe molar ratio of the tacrolimus is 0.13:0.93:23:0.03-0.15;
the reaction temperature is 90 ℃;
the reaction time is 5h;
the stirring speed during the reaction is 300rpm;
the centrifugation is carried out for 30min at 11500 r/m.
The application also provides a metal organic framework material prepared by the preparation method.
The application also provides application of the metal organic framework material to treatment of fungal keratitis.
The technical scheme of the application has the following advantages:
(1) The preparation method is simple, and the antifungal agent is coordinated with the metal-organic framework creatively by adopting a one-pot method, so that the more stable assembled organic metal framework N-PCN is obtained. The characterization and the function are explored by adjusting the input amount of antifungal drugs (such as NTM), and the optimal proportioning of N-PCN is finally determined. The method comprises the following steps: with meso-tetra (4-carboxyphenyl) porphyrin, zrOCl 2 ·8H 2 O and benzoic acid are used as materials of a metal organic framework, wherein meso-tetra (4-carboxyphenyl) porphyrin is used as a photosensitizer coordinated with metal Zr, so that the metal Zr has a large covalent component due to a coordination bond formed by high valence (6 valence) and a ligand, NTM forms coordination with an empty orbit of Zr, and the stability of the NTM-PCN metal framework is further enhanced.
(2) The application solves three problems of poor osmotic solubility, poor dispersibility and poor permeability of the antifungal drug, the solubility is improved by about 2 times, the permeability of the antifungal drug on cornea is enhanced due to positive charge (37+/-2 mv), the bioavailability is improved by about 25 times, and the antifungal drug stably exists in ultrapure water for more than three months. Therefore, the modified polysaccharide can be used as a universal carrier for delivering antifungal agents (mainly polyenes), and some load delivery (such as amphotericin AMB) is also carried out on other polyene antibiotics, thus having good prospect in the aspect of delivering antifungal agents.
(3) The application combines antifungal agent and photosensitizer to assemble new MOF, namely N-PCN, to realize photodynamic and chemical synergistic treatment, and has therapeutic effect of 1+1 greater than 2 on fungi (candida albicans is taken as an example). Experiments show that the targeting of natamycin on ergot-catalpol on the surface of fungi is utilized, so that N-PCN reaches an infected part and easily penetrates through a biomembrane, and is tightly combined with the surface of fungi, and active oxygen generated after laser irradiation is efficiently sterilized at the infected part, so that the synergistic sterilization of chemotherapy and photodynamic therapy is realized. Due to the synergistic effect of photodynamic and chemotherapy, the use amount of the two is reduced, effective bacteriostasis (bacteriostasis rate 95%) is achieved under the irradiation of 660nm laser with low power (0.2W) and short time (2 min), and meanwhile efficient sterilization (the bacteriostasis concentration of NTM is reduced from 5 mu g/mL to 0.5 mu g/mL) is achieved under the concentration of reduced NTM by 90%.
(4) The application can reach effective antibacterial concentration by low concentration and low illumination intensity and low illumination time, thereby greatly reducing dark toxicity and phototoxicity to normal cornea epithelial cells, having survival rate of more than 90 percent and realizing low PDT (general photodynamic damage to cells is larger).
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present application, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 is a TEM electron micrograph of the metal-organic framework material obtained in example 1 of the present application, wherein (A) and (B) are respectively at different scales;
FIG. 2 is a graph showing the results of the hydration particle size using DLS test of the metal organic framework material obtained in example 1 of the present application;
FIG. 3 shows the results of the drug loading rate test in test example 1 of the present application;
FIG. 4 shows the encapsulation efficiency test result in test example 1 of the present application;
FIG. 5 shows the results of particle diameter and potential tests in test example 1 of the present application, wherein (A) is N-PCN and (B) is PCN-N;
FIG. 6 is a graph showing the results of the Markov potential test in test example 1 of the present application;
FIG. 7 shows FT-IR results in test example 2 according to the application;
FIG. 8 is a UV-Vis result in test example 2 of the present application;
FIG. 9 is XRD results in test example 2 of the present application;
FIG. 10 shows BET results in test example 2 of the present application;
FIG. 11 shows the results of the solubility test in test example 3 of the present application, wherein (A) is PBS, (B) is NTM, (C) is H2TCPP, and (D) is N-PCN;
FIG. 12 shows the result of dissolution calculated in test example 3 of the present application;
FIG. 13 shows the change in PDI in the dispersibility test in test example 4 of the present application;
FIG. 14 is a photograph showing a dispersibility test in test example 4 of the present application;
FIG. 15 shows the results of the permeability test in test example 5 of the present application;
FIG. 16 shows the results of the changes in SIZE and PDI in the stability test of test example 6 according to the present application;
FIG. 17 is a TEM image in PBS of the stability test in test example 6 of the present application;
FIG. 18 shows the results of the sterilization performance test in test example 7 of the present application, wherein (A) is a photograph and (B) is a calculation result;
FIG. 19 is a TEM image after the sterilization performance test in test example 7 of the present application, wherein (A) is PBS, (B) is NTM, (C) is PCN, and (D) is N-PCN;
FIG. 20 is a TEM image of the surface of a fungus in the sterilization performance test of test example 7 according to the present application, wherein (A) is PBS, (B) is PCN, and (C) is N-PCN;
FIG. 21 is a graph showing cell viability at therapeutic concentrations in the absence of light in the toxicity test of test example 8 of the present application;
FIG. 22 shows the cell viability at therapeutic concentrations under light in the toxicity test of test example 8 of the present application;
FIG. 23 shows the results of apoptosis in the toxicity test of test example 8 of the present application, wherein (A) is PBS, (B) is NTM, (C) is PCN, and (D) is N-PCN;
FIG. 24 is a slit lamp test result in the body function verification test in test example 9 of the present application;
FIG. 25 shows the results of HE staining in the in vivo functional verification test in test example 9 of the present application, wherein (A) is PBS, (B) is NTM, (C) is PCN, and (D) is N-PCN.
Detailed Description
The following examples are provided for a better understanding of the present application and are not limited to the preferred embodiments described herein, but are not intended to limit the scope of the application, any product which is the same or similar to the present application, whether in light of the present teachings or in combination with other prior art features, falls within the scope of the present application.
The specific experimental procedures or conditions are not noted in the examples and may be followed by the operations or conditions of conventional experimental procedures described in the literature in this field.
Example 1
The embodiment provides a metal organic framework material, and the preparation method specifically comprises the following steps:
h2TCPP (100 mg,0.13 mmol), zrOCl 2 ·8H 2 O (300 mg,0.93 mmol), benzoic acid (2.8 g,23 mmol) and natamycin (100 mg,0.15 mol) were dissolved in 100mL dimethylformamide, stirred at 300rpm for 5h at 90℃and centrifuged (11500 r/m,30 min), then washed 3 times with pure dimethylformamide, and the resulting nanoparticles were suspended in DMF for use to obtain a metal organic framework material (N-PCN-100). The nanoparticles obtained in example 1, as shown in (A) and (B) in FIG. 1, have particle diameters of about 75-80nm in TEM at different magnification, and hydrated particle diameters of 90-100nm as shown in FIG. 2. The drug loading rate is 44% and the encapsulation rate is 60%.
Example 2
The embodiment provides a metal organic framework material, and the preparation method specifically comprises the following steps:
h2TCPP (100 mg,0.13 mmol), zrOCl 2 ·8H 2 O(300mg,0.93 mmol), benzoic acid (2.8 g,23 mmol) and natamycin (50 mg,0.075 mol) were dissolved in 100mL dimethylformamide, stirred at 300rpm for 5h at 90 ℃, centrifuged (11500 r/m,30 min), then washed 3 times with pure dimethylformamide, and the resulting nanoparticles were suspended in DMF for use to obtain a metal organic framework material (N-PCN-50).
Example 3
The embodiment provides a metal organic framework material, and the preparation method specifically comprises the following steps:
h2TCPP (100 mg,0.13 mmol), zrOCl 2 ·8H 2 O (300 mg,0.93 mmol), benzoic acid (2.8 g,23 mmol) and natamycin (20 mg,0.03 mol) were dissolved in 100mL dimethylformamide, stirred at 300rpm for 5h at 90℃and centrifuged (11500 r/m,30 min), then washed 3 times with pure dimethylformamide, and the resulting nanoparticles were suspended in DMF for use to obtain a metal organic framework material (N-PCN-20).
Example 4
The embodiment provides a metal organic framework material, and the preparation method specifically comprises the following steps:
h2TCPP (100 mg,0.13 mmol), zrOCl 2 ·8H 2 O (300 mg,0.93 mmol), benzoic acid (2.8 g,23 mmol) and amphotericin B (20 mg,0.02 mol) were dissolved in 100mL dimethylformamide, stirred at 300rpm for 5h at 90℃and centrifuged (11500 r/m,30 min), followed by washing 3 times with pure dimethylformamide, and the resulting nanoparticle was suspended in DMF for use to obtain the metal-organic framework material (A-PCN-20).
Comparative example 1
The difference between this comparative example and example 1 is that no natamycin was added and the preparation method is as follows:
h2TCPP (100 mg,0.13 mmol), zrOCl 2 ·8H 2 O (300 mg,0.93 mmol), benzoic acid (2.8 g,23 mmol) were dissolved in 100mL dimethylformamide, stirred at 300rpm for 5h at 90℃and centrifuged (11500 r/m,30 min), followed by washing 3 times with pure dimethylformamide, and the resulting nanoparticles were suspended in DMF for use to obtain a metal organic framework material (PCN).
Test example 1
The metal organic frame material obtained in examples 1-3 was referred to as N-PCN, and compared with nano-particles coated with NTM after PCN synthesis (abbreviated as PCN-N), and its basic properties were tested, as shown in fig. 3-6, wherein the drug loading in fig. 3, the encapsulation efficiency in fig. 4, the particle size and potential of N-PCN in fig. 5 (a), the particle size and potential of PCN-N, and the malvern potential in fig. 6, and it can be seen that the metal organic frame material N-PCN performance of the technical scheme of the present application is significantly superior to PCN-N.
Test example 2
This test example was analyzed by FT-IR, UV-Vis, XRD and BET for the metal-organic framework materials obtained in each example and comparative example 1, and natamycin, and the results are shown in FIGS. 7 to 10. It can be seen from FIG. 7 that natamycin itself has a peak around 1720nm, and when it is coordinated to metallic Zr through carboxyl groups, the disappearance of the peak shown by the carboxyl groups proves the coordination, and in FIG. 8, the disappearance of the peak around 300nm after the formation of the coordination can be seen through UV-Vis, and it can be confirmed that natamycin is actually present in the metallic organic frame material obtained in example 1 of the present application. It can be seen by XRD in fig. 9 that the coordination of natamycin does not alter the original crystal structure. As can be seen from the BET in FIG. 10, the N-PCN has a smaller pore size than PCN, N 2 The adsorption is reduced, which indicates that more coordination occupies the pore canal and the pore diameter is reduced.
Test example 3
In this test example, the metal-organic framework material obtained in example 1 of the present application was compared with free NTM and H2TCPP, as shown in fig. 11, where (a) in fig. 11 is PBS, (B) is NTM, (C) is H2TCPP, (D) is N-PCN, that is, example 1, PBS is transparent and clear, as a control group, and NTM and H2TCPP of equal mass contained in N-PCN are dissolved in 2mL of PBS, respectively, and it is found that NTM presents suspension, H2TCPP also presents insoluble state, both stand still, and N-PCN remains well dispersed, and it can be visually reflected that NTM presents suspension of equal amount and well dispersed in nano state, and it can be seen that the specific value of improvement is calculated in fig. 12, and the metal-organic framework material obtained in the embodiment of the present application increases the solubility of NTM by about 2 times.
Test example 4
In this test example, compared with comparative example 1, the metal-organic frame material obtained in each example of the present application has a PDI of less than 0.1 within 30 days as shown in fig. 13, and the metal-organic frame material obtained in each example of the present application maintains good dispersibility in ultrapure water for 3 months as shown in fig. 14, whereas the metal-organic frame material in comparative example 1, to which natamycin is not added, has not been able to maintain a dispersed state, which indicates that the coordination of natamycin improves the stability of the metal-organic frame material.
Test example 5
In this test example, compared with free antibiotic NTM, the metal organic frame material obtained in example 1 of the present application was subjected to anesthesia treatment, and the cornea was sacrificed, and an in vitro diffusion cell experiment was performed, wherein 1 ml of liquid was added to a donor cell, and samples were taken at different time points, and as a result, as shown in fig. 15, it was seen that the bioavailability was greatly improved.
Test example 6
In this test example, the metal-organic framework material obtained in example 1 of the present application was dispersed in ultrapure water, and as shown in fig. 16, the Size was not significantly changed within 7 days, and the PDI was kept below 0.1, indicating that the metal-organic framework material obtained in the present application was highly stable in ultrapure water and suitable for storage. As shown in fig. 17, in PBS, it can be seen by taking a TEM that the metal frame gradually disintegrates due to the ion exchange principle in PBS, gradually releasing the N-PCN component.
Test example 7
The test example is a sterilization performance test.
The metal organic frame materials obtained in example 1 and comparative example 1 were subjected to plate counting by diluting the coated plates after co-incubation with fungi (candida albicans) in 96-well plates for 24 hours, the results are shown in fig. 18, wherein (a) is a photograph, statistical data is processed into bar graphs, the bar graphs are shown in (B), bacterial solutions after co-incubation for 24 hours are diluted and subjected to TEM as shown in fig. 19, wherein (a) in fig. 19 is PBS, (B) is NTM, (C) is PCN, i.e., comparative example 1, and (D) is N-PCN, i.e., example 1. Can pass throughFig. 18 and 19 show that the N-PCN obtained in example 1 has a better bacteriostatic effect than the free NTM and PCN at the same dose. FIG. 20 illustrates the mechanism by which N-PCN has a better bacteriostatic effect, wherein (A) in FIG. 20 is PBS, (B) is PCN, comparative example 1, and (C) is N-PCN, example 1, more nanoparticles are targeted to bind to the fungal surface due to the specific binding of NTM in N-PCN to the fungal surface. The specific method of the test example is as follows: activating Candida albicans on culture medium twice, diluting the strain to 10 with turbidimetric tube containing NaCl solution on turbidimetric apparatus 7 (turbidity of 0.7), and diluted to 10 with 1640 medium 5 N-PCN was diluted to a NTM concentration of 0.5. Mu.g/mL and incubated in 96-well plates for 24 hours to a dilution of 10 3 Plating, culturing for 36 hours, and counting the plates to obtain corresponding sterilization results. In each of these figures, PBS represents PBS buffer solution, NTM represents free natamycin, PCN means that PCN without NTM is irradiated with 660nm laser light of 0.2W for 2min, and N-PCN means that the metal-organic framework material N-PCN obtained in example 1 is irradiated with 660nm laser light of 0.2W for 2min.
Test example 8
This test example is a toxicity test for the metal organic framework material obtained in example 1, all cells were cultured in F12 medium containing 10% calf serum and placed at 37℃and 95% O 2 And 5% carbon dioxide incubator. These cells were subcultured by centrifugation (1000 rpm,3 min) with 0.25% trypsin. HCEC cells (corneal epithelial cells) were detected using the standard CCK-8 method. Cells were cultured in 96-well plates (90. Mu.L/well, 8.0X104 cells/mL) in an incubator at 37℃for 12 hours. NAT-PCN NPs (10. Mu.L) were then dispersed in fresh medium at various concentrations (0, 0.25, 0.5, 1, 2, 4, 6. Mu.g/mL) for co-incubation for 24 hours. Thereafter, the medium was removed, washed three times with PBS, incubated with CCK-8 solution (100. Mu.L, 10-fold diluted with medium) for 4 hours, absorbance detection was performed at 450nm with a microplate reader at 2h, 3h, 4h, respectively, and recorded. As a result, as shown in fig. 21 and 22, in fig. 21, the cell viability at the therapeutic concentration was more than 90% in the absence of light; in FIG. 22, after the irradiation, the irradiation intensities of 200mw and 300mw were set to 3min and less than 3minThe cell survival rate can be maintained to be more than 80% in all time, and gradually decreases with the time. FIG. 23 shows a flow-through apoptosis experiment, wherein the biocompatibility was compared by comparing the sum of Q2+Q3, wherein (A) in FIG. 23 is PBS, (B) is NTM, (C) is PCN, which is comparative example 1, and (D) is N-PCN, which is example 1. It can be seen that the apoptosis rate (3.52+2.09)/100 of N-PCN is less than 10%, and is consistent with the CCK8 result, the N-PCN is further proved to have good biocompatibility, and low PDT sterilization is realized in a safe range.
Test example 9
The test example is an in-vivo functional verification test. FIG. 24 shows slit lamp data, and the results of observation of the cornea on days 0, 1, 3 and 5, respectively, show that the metal-organic framework material N-PCN obtained in example 1 has good healing effect on day 5, and shows corneal glipting. Fig. 25 shows HE staining experiments, in which (a) in fig. 25 is PBS, (B) is NTM, (C) is PCN, that is, comparative example 1, and (D) is N-PCN, that is, example 1, and it can be seen that the cornea treated with the metal-organic framework material N-PCN obtained in example 1 has no obvious thickening or inflammation, which proves that the cornea has a good therapeutic effect in vivo.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. While still being apparent from variations or modifications that may be made by those skilled in the art are within the scope of the application.
Claims (4)
1. A method for preparing a metal organic framework material for treating fungal keratitis, which is characterized by comprising the following steps:
s1: meso-tetra (4-carboxyphenyl) porphyrin and ZrOCl 2 ·8H 2 Dissolving O, benzoic acid and polyene antibiotics in dimethylformamide for mixed reaction, centrifuging and taking solid to obtain a crude product;
s2: washing the crude product with dimethylformamide to obtain the metal organic framework material.
2. A process according to claim 1, wherein the polyene antibiotic is natamycin.
3. The method according to claim 2, wherein in step S1, the meso-tetra (4-carboxyphenyl) porphyrin, zrOCl 2 ·8H 2 The mol ratio of O, benzoic acid and natamycin is 0.13:0.93:23:0.03-0.15;
the reaction temperature is 90 ℃;
the reaction time is 5h;
the stirring speed during the reaction is 300rpm;
the centrifugation is carried out for 30min at 11500 r/m.
4. A metal organic framework material prepared by the preparation method of any one of claims 1 to 3.
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