Metal organic framework nanoparticles for oral protein administration and preparation method of metal organic framework nanoparticles
A metal-organic framework and nanoparticle technology, applied in the field of pharmacy, can solve the problems of low protein oral bioavailability and complex product preparation process, and achieve good biocompatibility, slow and controlled release kinetics, low price, and convenient protein The effect of oral administration
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[0029] Example 1 Preparation and Characterization of Oral Insulin Metal Organic Frame Nanoparticles (T @ i @ u)
[0030] UIO-68-NH by solvent heat 2 : Amino-TPDC (10.5 mg, 0.031mmol) and Zrocl 2 · 8h 2 O (8.3 mg, 0.026 mmol) was dissolved in 2 ml DMF. The mixture was so late until it clarified and transparent. 40 μl of acetic acid was added to the mixture, followed by heating at 90 ° C for 6 hours. The mixture was centrifuged at 11000 rpm and washed 3 times with DMF 3 to obtain purified UIO-68-NH. 2 . Continue to centrifuge at 11000 rpm 3 times to exchange DMF into DDH 2 O.
[0031] Under the conditions of stirring at room temperature, 50-300 μl of insulin solution (20 mg / ml, pH = 2.0, dilute hydrochloric acid is solvent) Add to 4 ml of new synthetic UIO-68-NH 2 (0.25 mg / mL) Aqueous solution was stirred for 12 hours to obtain insulin @ metal organic frame nanoparticles (I @ u). Subsequently, 40 μl of the anti-ferrous solution (5 mg / ml) was added to the above solution and st...
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[0033] Example 2 Insulin activity detection
[0034] To verify that insulin still has a relatively biologically active during the preparation of nano-preparation and nano-formulation, in this example, I @ u and t @ i @ u at 37 will be prepared according to the method in Example 1 The 1 ml of PBS buffer was immersed in ° C for 24 hours and the supernatant was separated by a low temperature overspeed (11000 rpm). The amount of insulin released in the supernatant was determined using a BCA kit, and then the released insulin (5 Iu / kg) and the released insulin (5 Iu / kg) were given overnight fastening SD rats. The blood glucose concentration of rats at different time points was measured. Such as image 3 As shown, it is shown that the process of insulin metal organic frame nanoparticles prepared in the present invention does not impair the biological activity of insulin.
Example Embodiment
[0035] Example 3 T @ i @ u on insulin protection characteristics
[0036] In order to determine UIO-68-NH 2 Whether it can protect insulin is subject to digestive enzymes, and I @ U and T @ i @ u in Example 1 will be incubated at 37 ° C in HBSS buffer containing trypsin (1 mg / ml) in HBSS buffer in Example 1. The aliquot (100 μl) was taken out at a specific time interval, and 200 μl of DMSO containing 0.1% trifluoroacetic acid was added to terminate the enzymatic reaction, and the concentration of insulin in the solution was determined using an ELISA kit. Such as Figure 4 As shown, I @ u and t @ i @ u can protect insulin in 2 hours to damage insulin.
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