title: Development of Salvianolic acid B-Tanshinone II A-Glycyrrhetinic acid compound liposomes: formulation optimization and its effects on proliferation of hepatic stellate cells.
abstract: The aim of this study was to systematically optimize and characterize the co-encapsulation process of Salvianolic acid B (Sal B), Tanshinone II A (TSN) and Glycyrrhetinic acid (GA) into liposomes. The liposomes (GTS-lip) were prepared using film hydration method combined with probe sonication to encapsulate two hydrophobic components (TSN and GA), and using pH gradient method to load hydrophilic component Sal B. The concentration of encapsulated drugs was measured by a reversed phase high performance liquid chromatography (RP-HPLC) method. Systematic optimization of encapsulation process was performed using single factor test, orthogonal test in combination with Box-Behnken Design. Optimum conditions are as follows: ratio of GA to lipid (w/w)=0.08, ratio of Sal B to lipid (w/w)=0.12 and pH of buffer=3.3. Based on the conditions mentioned above, encapsulation efficiency of Sal B, TSN and GA reached target levels: (96.03 ± 0.28)%, (80.63 ± 0.91)% and (88.56 ± 0.17)%, respectively. The GTS-lip had a unimodal size-distribution and a mean diameter of 191.3 ± 6.31 nm. Morphology determination of the GTS-lip indicated that the liposomes were spherical, and there was no free drug crystal in the visual field of transmission electron microscopy. Also, the ζ potential of GTS-lip was detected to be -11.6 ± 0.35 mV. In vitro release investigation of GTS-lip suggested that the release rate of GTS-lip significantly decreased compared to drug solution. The accumulative release percentage of TSN, GA and Sal B were 10% in 36 h, 4% in 36 h and 77% in 24 h. Meanwhile, GTS-lip exhibited definite activity on proliferative inhibition of hepatic stellate cells (HSC). GTS-lip decreased the viability of the HSC to higher than 75% at two high drug concentration groups in 24h. At the same time, GTS-lip of two low drug concentration groups increased the inhibition rates by 2.3 folds and 1.9 folds separately at 48 h compared to 24h. By contrast, inhibition activity of G-T-S solution group showed less change between 48 h and 24 h. The prolonged and enhanced activity in 48 h which GTS-lip group manifested might contribute to its sustained release effect.
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Jiahao
authors.initials: J
authors.lastname: Lin
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Xiuli
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Wu
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Jundong
authors.initials: J
authors.lastname: Dai
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Huida
authors.initials: H
authors.lastname: Guan
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Weiyi
authors.initials: W
authors.lastname: Cao
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China.
authors.forename: Liangying
authors.initials: L
authors.lastname: He
authors.affiliation: Department of Chinese Pharmaceutics, School of Chinese Materia Medica, Eastern Campus, Beijing University of Chinese Medicine, Beijing 100102, PR China. Electronic address: yurong.wang@163.com.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Wang
mesh_terms: D045784:Abietanes / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D001572:Benzofurans / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D003692:Delayed-Action Preparations; D006034:Glycyrrhetinic Acid / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008081:Liposomes; D046529:Microscopy, Electron, Transmission; D010316:Particle Size; D013997:Time Factors; D053630:Depsides
chemical_list: D045784:Abietanes; D001572:Benzofurans; D003692:Delayed-Action Preparations; D006034:Glycyrrhetinic Acid; D008081:Liposomes; C021751:tanshinone; C076944:salvianolic acid B; D053630:Depsides
keywords: Co-delivery; Glycyrrhetinic acid; Glycyrrhetinic acid (PubChem CID: 10114); Hepatic stellate cells; Liposomes; Salvianolic acid B; Salvianolic acid B (PubChem CID: 13991587); Tanshinone II A; Tanshinone II A (PubChem CID: 164676)