Preparation Of Lycopene Inclusion Complex Via Hot-melt Extrusion

OBJECTIVE:To improve the solubility and dissolution of lycopene,and increase it stability,the hot melt extrusion techology was employed to prepare lycopene Hydroxypropyl-β-cyclodextrin inclusion compound;The effects of lycopene and its preparation on proliferation apoptosis of human prostate PC-3 cancer cell line were investigated;The absorption mechanism in vitro and its pharmacokinetics in vivo were also studied.METHODS:High performance liquid chromatography was established to determined the content of lycopene and its preparation;Hot melt extrusion was used to prepared lycopene hydroxypropyl-β-cyclodextrin inclusion compound,and its solubility and dissolution were measured.The inclusion complex was characterized by differential scanning calorimetry,Fourier transform infrared spectroscopy and X-ray powder diffraction;The powder direct compression was used to make tablets,the content and the dissolution were selected as indicators to assess the stability of tablet under the condition of 4℃,25℃ and 40℃;The Caco-2 cell monolayer model was established to investigate the absorption of lycopene and its preparation.The method of high performance liquid chromatography for the determination of lycopene in Beagle plasma was carried out.Taking The commercially available lycopene tablets as the reference(reference tablets),the pharmacokinetics and bioavailability of homemade lycopene inclusion complex tablets were investigated;The MTT method was used to observe the effects of lycopene on the proliferation of PC-3 cells,and cell apoptosis were invesstigated by flow cytometry.The long-term dynamic live cell observation system was also used to observe cell proliferation and apoptosis.RESULTS:The results of method validation suggested that HPLC method used to determine the content of lycopene was fit for the requirement of determination;The formation of lycopene inclusion complex was confirmed by differential scanning calorimetry,Fourier transform infrared spectroscopy and X-ray powder diffraction analysis,and the dissolution and solubility have been greatly improved;In the stability experiment,under the condition of 4℃,the content and dissolution of lycopene remained stable within three months,however,under the condition of 25℃,the content decreased as the time increased,but the dissolution was not affected,the content and dissolution decreased significantly at 40℃ conditions;Lycopene can inhibit the proliferation and induce apoptosis of PC-3 cells showing time and dose dependent manner;The results of Caco-2 cell monolayer transport experiment suggested that the uptake of lycopene inclusion complex is better than raw materials,showing time and dose dependent manner,the higher amounts absorbed when lycopene is added to the apical chamber compared to the basolateral one,and the absorption pattern suggested that lycopene was passively transported;The plasma concentration-time data were analyzed by non-compartmental methods using DAS2.1 software,the Cmax and AUC0-t of homemade lycopene inclusion complex tablets were 4.88 and 2.56 times higher,respectively,compared with those of reference tablets,and the relative bioavailability of lycopene of homemade lycopene inclusion complex tablets was(313.08±40.5)%.CONCLUSION:With the hydroxypropyl-β-cyclodextrin as the inclusion material,PEG6000 as the plasticizer,hot melt extrusion was used to prepare lycopene inclusion complex,which greatly improved the dissolution and solubility.The preparation method is simple and can be continuously produced in large scale;The results showed that the absorption of lycopene inclusion complexes on Caco-2 monolayer cells was superior to that of raw lycopene,and the pharmacokinetic experiments showed that,compared with commercially available lycopene tablets,the relatively bioavailability significantly increased;The lycopene can inhibit the proliferation and induce apoptosis of PC-3 cells,and it has a certain time-dependent dose-dependent relationship.