Preparation And Effectiveness Of Solubilized Naringenin Eye Drops

ObjectiveNaringenin(NAR),comprehensive pharmacological study shows that naringenin have rich pharmacological activity,such as anti-oxidant effects.Naringenin has shown some efficacy in eye related diseases,including inhibition of retinal damage,diabetic retinopathy,uveitis,corneal neovascularization,etc.The poor water solubility of naringenin has limited its application in ophthalmology,and some important parameters such as stability and other efficacy have not been well solved.Therefore,the preparation of an effective solubilization of naringenin eye ophthalmic preparation has important significance.In this study,two kinds of excipients were used to prepare solubilized naringenin eye drops,and their characterization and pharmacodynamics were evaluated.Methods1.The solubilized naringenin eye drops were prepared by solvent evaporation hydration method with Kollidon(?) 17PF(17PF)as excipients,the complexation efficiency and other indexes were determined;The storage stability was investigated at4℃ and 25℃ respectively;The anti-oxidation activity was measured by ABTS method;The ocular local irritation was evaluated by pathological examination,and the cytotoxicity of eye drops was tested by MTT assay;The in vitro release of naringenin eye drops was measured by dialysis membranes;The ocular absorption and distribution in rabbits of naringenin eye drops were measured by HPLC;The ocular surface inflammation model was established to evaluate the anti inflammatory activity of eye drops.2.Dipotassium glycyrrhetate(DG)was used as excipient to prepare dipotassium glycyrrhizinate-solubilized naringenin micellar eye drops,the encapsulation efficiency and others were measured;Anti-oxidation activity was tested by ABTS method;Local irritation of eyes was evaluated by ocular comprehensive scoring method;The absorption of the eye drops in the cornea of mice was detected by fluorescence labeling with Cou-6.3.Balb/C mice were treated with 0.2% benzalkonium chloride(BAC)to construct dry eye model.The mice were divided into different groups randomly.Different drug was given while the Benzalkonium chloride was maintained,compared with blank group;Fluorescein staining and tiger red staining of cornea were observed by slit-lamp;And the tear secretion and corneal sensitivity were measured;The changes of corneal epithelium/endothelial microstructure were observed by SEM;Corneal pathology was evaluated by HE staining;WB was performed to evaluate the expression profilesl of High Mobility Group B1(HMGB1);The expressions of IL-6 and IL-1β were detected by ELISA.Results1.The complexation efficiency of 17 PF solubilized naringenin eye drops was98.51±0.86%,the particle size was 6.73±0.42 nm,and the storage stability was good.17 PF solubilized naringenin increased the antioxidant activity of naringenin and the membrane permeability of naringenin,showed good in vitro cell tolerance and no ocular local irritation;The ocular absorption was greatly improved compared with naringenin solution.17PF-NAR significantly increased the anti-inflammatory activity of NAR in rabbit eyes.2.The encapsulation efficiency of DG-NAR was nearly 100.00%,the particle size was 24.75±0.52 nm.The antioxidation activity of DG-NAR was higher than that of NAR and DG solution.The safety of the eye is good and the absorption in the cornea is greatly improved.3.The corneal sensitivity and tear secretion of the dry eye mice were reduce.Compared with the mice of PBS group,we found that the treatment degree of other groups was improved,DG-NAR group improved the recovery of dry eye effectively(p<0.05);The level of inflammatory factors in cornea of the DG-NAR eye drop group decreased,and the expression of HMGB1-related protein in DG-NAR eye drops group was lower than that in other groups(p<0.05).ConclusionBoth 17 PF and DG can significantly solubilize naringenin.The prepared eye drops have appropriate characterization and properties.Naringenin has a good anti-inflammatory effect,17PF-NAR eye drops play an important role in the treatment of ocular inflammation;DG-NAR eye drops is essential in regulating the signal pathway of HMGB1 in dry eyes,the solubilized naringenin eye drops based on HMGB1 inhibitor-DG play a role of multi-target regulating HMGB1 signal pathway to prevent and treat dry eye effectively.