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ObjectiveTo prepare oral nanoemulsions encapsulating essential oil from Alpinia zerumbet fructus（EOFAZ） and to investigate its pro-absorption effect in vitro and distribution in vivo. MethodThe proteoglycan conjugate polysaccharides of vinegar-processed Bupleuri Radix-bovine serum albumin（VBCP-BSA） was prepared by Maillard reaction of VBCP and BSA. Taking VBCP-BSA as emulsifier， vitamin B12（VB12） as absorption enhancer， and medium chain triglycerides mixed with EOFAZ as oil phase， the nanoemulsions loaded with EOFAZ was prepared by high energy emulsification method. The particle size， particle size distribution， surface Zeta potential， EOFAZ content and appearance and morphology of the nanoemulsions were characterized， and fluorescein tracer method was used to investigate the absorption effect of fluorescein-labeled EOFAZ nanoemulsions in vitro and their distribution in vivo. ResultVBCP-BSA was formed by Maillard reaction for 48 h with high grafting rate. Using VBCP-BSA as emulsifier， the homogeneous pink nanoemulsions was prepared and denoted as EOFAZ@VBCP-BSA/VB12. The particle size of the nanoemulsions was less than 100 nm and the particle size distribution was uniform. The surface of the nanoemulsions was a weak negative charge， and the shape was spherical. The encapsulation rate of the nanoemulsions for EOFAZ was greater than 80%， which had a good absorption effect in vitro and could enhance liver accumulation after oral administration. ConclusionThe designed proteoglycan nanoemulsions can effectively load EOFAZ， promote oral absorption and enhance liver distribution， which can provide experimental basis for the development of oral EOFAZ liver protection preparations.
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Objective:To construct the targeting evaluation method of traditional Chinese medicine （TCM） preparations based on supramolecular Qi chromatography theory， and to study the liver targeting effect of Bupleuri Radix on Pien Tze Huang. Method:The molecular connectivity index （MCI） was used to analyze the characteristics of imprinted template and liver targeting tendency of TCM mainly attributed to liver meridian and components of Pien Tze Huang， and combined with target dynamics and total statistical moment principle， aimed at the independent action characteristics of multi-component imprinted template of TCM， a method for evaluating the targeting of TCM preparations was established. Hepatoma rats in Pien Tze Huang group， Bupleuri Radix<italic> </italic>group， Pien Tze Huang+Bupleuri Radix group and blank group were tested and verified. Result:After the average value of MCI of TCM mainly attributed to liver meridian was deducted， the MCI similarity between Pien Tze Huang group and Bupleuri Radix group was 0.376 8， Pien Tze Huang+Bupleuri Radix group and Bupleuri Radix group was 0.988 2， so it was predicted that Bupleuri Radix could enhance the liver targeting of Pien Tze Huang. A system for evaluating the targeting of TCM compounds was established， including relative total uptake efficiency （RUE<sub>T</sub>）， relative total concentration （RC<sub>T</sub>）， relative imprinted tendency （RIT<sub>T</sub>） and relative imprinted variance （RIV<sub>T</sub>）. The RUE<sub>T</sub> and RC<sub>T</sub> of liver were the highest in all tissues （RUE<sub>T</sub>=1.88>1，RC<sub>T</sub><italic>=</italic>2.30>1）， and the corresponding values of other tissues were all <1， indicating that Pien Tze Huang combined with Bupleuri Radix could increase its distribution in liver and enhance liver targeting. Except for plasma， the RIT<sub>T</sub> and RIV<sub>T</sub> of other tissues fluctuated around 1.0， indicating that targeted modification did not change imprinted tendency of Pien Tze Huang and had no significant effect on the types of components. Conclusion:Under the guidance of supramolecular Qi chromatography theory， a targeting evaluation parameter system can be established to characterize the multi-component imprinted effect of TCM preparations by MCI and total statistical moment parameters， so as to realize the evaluation of targeting of TCM preparations. The addition of Bupleuri Radix can increase the liver targeting of Pien Tze Huang.
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Objective To construct three kinds of doxorubicin liposomes modified with cholesterol-galactose ligand by lipase-catalyzed method and compare their characteristic of pharmacokinetics and tissue distribution in vivo. Methods Three types of cholesterol-galactose ligands, CHS-C8-GalNAc, CHS-C8-GAL, and CHS-C8-LA were synthetized by lipase-catalyzed method in nonaqueous phase. The structure characterizations of products were obtained by MS and NMR. Conventional liposomes (CL DOX) and ligand-coupled liposomes (NGal-LP DOX, Gal-LP DOX, and LA-LP DOX) were prepared by thin film dispersion-ammonium sulphate gradient method. Structure-activity relationship between asialoglycoprotein receptor (ASGPr) and the chemical structure of the glycolipids was explored through the pharmacokinetics and tissue distribution parameters of ligand-coupled liposomes in vivo. Results The desired compounds with a high yield of above 90% were confirmed by MS and NMR. The liposomes average size was lower than 90 nm, polymer dispersity index was lower than 0.1, encapsulation efficiency was greater than 99%, leakage rat was lower than 5% with 24 h, and zeta potential closed to zero. The affinity of the three ligand molecules to liver was the following order: CHS-C8-GalNAc > CHS-C8-LA > CHS-C8-Gal. However, only the liposomes modified with CHS-C8-GalNAc could significantly be inhibited by the preinjection of asialofetuin for hepatic uptake rate (P 0.05). Conclusion The ligand with N-acetylgalactosamine residue showed high targeting efficiency for hepatocytes, while the ligand with D-galactose (Gal) or lactitol residue could competitive bind with Gal particle receptor on kupffer cells.
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Based on the research of active liver targeting liposomes mediated by glycyrrhetinic acid ligand at home and abroad, this paper focuses on the liver targeting effect of liposomes mediated with 18-GA-Gly, a kind of glycyrrhetinic acid ligand. salvianolic acid B(Sal B)-tanshinone ⅡA (TSN)liposomes mediated by 18-GA-Gly as well as the liposomes with unmodified ligands were prepared by film dispersion-high pressure homogenization method, and then the particle size, potential, encapsulation efficiency and ligand binding rate were detected. Plasma samples of the heart, liver, spleen, lung and kidney tissues were taken at different time points after tail vein injections. The contents of Sal B and TSN in each sample were determined with UPLC methods and the liver targeting effect of 18-GA-Gly ligands was evaluated. The results showed that the particle size, potential, encapsulation efficiency and ligand binding rate met the basic requirements; in vivo targeting investigation results showed no difference between GLY-TS-Lip group and TS-Lip group. The liposomes mediated by glycyrrhetinic acid derivative ligand 18-GA-Gly can increase the peak concentration of Sal B and TSN in liver, but showed no significant liver targeting effect.
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To study the tissue distribution of galactosyl daphnoretin liposomes in rats. At the dose of 10 mg•kg⁻¹, daphnoretin solution, daphnoretin liposomes, and galactosyl daphnoretin liposomes were administered to healthy SD rats via tail vein injection. The blood and tissue of heart, liver, spleen, lung, kidney, stomach, small intestine, brain and thymus were collected at 5, 15, 30, 45, 60, 120, 240, 360 min after administration. The concentrations of daphnoretin in plasma and tissue samples were determined by HPLC. The results showed that galactosyl daphnoretin liposomes group had the highest concentration of daphnoretin in liver of unit weight at different time points; and at all of the time points, the target index DTI values of galactosyl daphnoretin liposomes to liver were greater than that of daphnoretin liposomes. Compared with daphnoretin solution, the AUC0-6 and Cmax of galactosyl daphnoretin liposomes in liver were 2.23, 5.22 times, respectively. This indicated that galactosyl daphnoretin liposomes can be concentrated at liver, with a significant liver targeting effect.
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OBJECTIVE:To prepare folic acid(FA)-loaded vincristine(VCR)nano liposome(VCR-nLip-FA)and to study its effects on human liver and lung cancer cells. METHODS:VCR-nLip-FA was prepared by ammonium sulfate gradient method,and particle size,Zeta-potential,encapsulation rate and release rate were investigated. Taking human liver cancer HepG2 cells and lung cancer A549 cells as example,uptake rate and inhibitory effect in vitro (5-80 μg/ml) were compared between VCR-nLip-FA and VCR-nLip. RESULTS:The particle size distribution,average particle size,average Zeta-potential,average encapsulation rate and 24 h accumulative release rate of VCR-nLip-FA were 98.1-159.0 nm,132.2 nm,-40.1 mV,(86.6±3.5)%(n=4)and(42.2± 2.6)%. Compared with VCR-nLip,there was no statistical significance in uptake rate of A549 cells to VCR-nLip-FA and inhibitory effect of VCR-nLip-FA on A549 cell viability (P>0.05);uptake rate of HepG2 cells to VCR-nLip-FA and inhibitory effect of VCR-nLip-FA on HepG2 cell viability increased significantly (P<0.01),in dose-dependent manner. CONCLUSIONS:Prepared VCR-nLip-FA can target anti-tumor drug to HepG2 cells efficiently,and highly inhibit the growth of HepG2 cells. But it has no higher effects on A549 cells.
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The objective of this study was to prepare cubosomal nanoparticles containing a hydrophilic anticancer drug 5-fluorouracil (5-FU) for liver targeting. Cubosomal dispersions were prepared by disrupting a cubic gel phase of monoolein and water in the presence of Poloxamer 407 as a stabilizer. Cubosomes loaded with 5-FU were characterized in vitro and in vivo. In vitro, 5-FU-loaded cubosomes entrapped 31.21% drug and revealed nanometer-sized particles with a narrow particle size distribution. In vitro 5-FU release from cubosomes exhibited a phase of rapid release of about half of the entrapped drug during the first hour, followed by a relatively slower drug release as compared to 5-FU solution. In vivo biodistribution experiments indicated that the cubosomal formulation significantly (P<0.05) increased 5-FU liver concentration, a value approximately 5-fold greater than that observed with a 5-FU solution. However, serum serological results and histopathological findings revealed greater hepatocellular damage in rats treated with cubosomal formulation. These results demonstrate the successful development of cubosomal nanoparticles containing 5-FU for liver targeting. However, further studies are required to evaluate hepatotoxicity and in vivo antitumor activity of lower doses of 5-FU cubosomal formulation in treatment of liver cancer.
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Objective To prepare liposomes of glycyrrhizic acid, and evaluate its liver targeting property in mice. Methods The liposomes were prepared with conventional rotary-evaporated film-ultrasonication method.The liposomes were injected into the mouse tail vein, and the concentration of glycyrrhizic acid was detected by RP-HPLC.The glycyrrhiz-ic acid injection was taken as control.The targeted indicators, including the relative tissue exposure ( re ) , targeting effi-ciency (te), and index of peak concentration ratio (Ce), were used to evaluate the liver targeting property.Results The re was 1.4, Te of the liposomes was 0.092, and te of the injection was 0.059.The Ce of the liver was 1.59, and the Ce of the blood was 0.99.Conclusion Compared with glycyrrhizic acid injection, the glycyrrhizic acid liposomes show good liv-er-targeting property.
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Objective: To investigate the antitumor activity of pH-sensitive liposomes (pH-LPC-lips) loaded with lactosyl-norcantharitin (Lac-NCTD) phospholipids complex in vitro and in vivo, and the liver targeting in mice. Methods: Using blank liposomes (blank-lips and blank-pH-lips) as control, the MTT assay was used to study the cytotoxic effects of Lac-NCTD and its liposomes (Lac-lips and pH-LPC-lips) on human hepatoma carcinoma cells HepG2. HPLC assay was used to evaluate the uptake of Lac-NCTD and its liposomes in HepG2. In vivo antitumor activity of Lac-NCTD and its liposomes were evaluated in mice bearing H22 liver tumors. The hepatocyte specificity of near-infrared fluorescence dye (Cy7)-labeled pH-LPC-lips in H22 tumor-bearing mice was monitored through NIR fluorescence real-time tumor imaging instrument. Results: The pH-LPC-lips demonstrated stronger cytotoxicity against tumor cells HepG2 and easily permeated the cell membrane, compared with Lac-NCTD and Lac-lips. The results of antitumor activity in vivo showed that pH-LPC-lips displayed best tumor inhibitory effect. The optical imaging results indicated that Cy7-labeled pH-LPC-lips showed excellent hepatocyte specificity in H22 tumor-bearing mice, which could reduced the side effect, and increased the antitumor activity. Conclusion: The pH-LPC-lips could take the initiative to release at the tumor site and showed the liver-targeting. As a result, the preparation could be regarded as novel liver-targeting agent which has better antitumor effect.
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Objective To investigate the hepatocyte targeted specific property of galactosylated chitosan-graft-polyethyleneimine (GC-PEI)/DNA complexes in vitro and in vivo.Methods With the plasmid expressing enhanced green fluorescent protein (pEGFP-C1) as the reporter gene,the formation of GC-PEI/DNA complexes was induced to self-assemble in 0.01 mol/L phosphate buffered saline(PBS),150 mmol/L NaCl,or 5% glucose solution (GS).The complexes were characterized by the particle size,Zeta potential,DNA binding and protection capacity,and further tested for cytotoxicity and hepatocyte targeted degradation of DNaseⅠand the serum,which presented as a well-formed sphere or compacted nucleocapsid structure at a diameter of 50-200 nm.The GC-PEI copolymer showed no obvious toxicity in the tested cell lines.Acute toxicity assay revealed that the mice grew well in 2 weeks with GC-PEI dosage from 50 to 300 μg.The assay by flow cytometry and fluorescent microscope showed that the transfection efficiency in hepatocyte lines (L02,QSG7701/core) was higher than that in non-hepatocyte lines (SGC7901,HBE) in vitro.In vivo,the GFP was obviously expressed in the liver tissue and not expressed in other organs 48 h after the transfection.Conclusion GC-PEI copolymer may carry the exogenous gene specifically to hepatocytes in vitro and in vivo,which has very good liver targeted specific property.
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Objective: To investigate the anticancer activity of the novel lactosyl-norcantharidin nanoparticles (Lac-NCTD-NPs) in vivo and in vitro. Methods: The MTT method was used to study the cytotoxic effects of Lac-NCTD and Lac-NCTD-NPs on HepG2, SMMC-7721, and SGC-7901 cell lines for 12 and 48 h, respectively, and the inhibitory effects of Gal-FBS; Lac-NCTD accumulated in SMMC-7721 cells was assayed by HPLC; The in vivo anticancer activity was evaluated by the tumor-growth inhibition in H22 tumor bearing mice. Results: The in vitro studies showed that the cytotoxic effects of Lac-NCTD-NPs against HepG2 and SMMC-7721 cells were the most powerful, as well as the IC 50 was the lowest, then Lac-NCTD, and they were inhibited remarkably by Gal-FBS; As for SGC-7901 cell line, the cytotoxic effects of Lac-NCTD-NPs and Lac-NCTD were not stronger than that of NCTD, and Gal-FBS had no influence on them at all; The amount of Lac-NCTD accumulated in SMMC-7721 cells was 3. 89 μg (7.02×10-3 μmol, 1×106 cell) after treatment for 12 h; The results of the antitumor activity in vivo suggested that the inhibitory rate of Lac-NCTD-NPs on tumor weight was 63.9%, which was significantly higher than that of NCTD and Lac-NCTD groups at the same molar concentration. Conclusion: The tumor-growth is inhibited effectively by Lac-NCTD-NPs which may be a kind of novel liver-targeting agents and could strongly inhibit the tumor-growth.
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OBJECTIVE:To prepare glycyrrhetic acid derivatives-modified norcantharidin(NC)liposome(GDNL)and study its liver-targeting property.METHODS:Gal-GAOSt targeting molecules were synthesized to modify NC and prepare GDNL,with the parameters such as the entrapment efficiency and particle diameter,etc.investigated.Mice were enrolled to be injected with GDNL and NC water solution,respectively via vena caudalis followed by determination of NC concentration in different tissues to compute the targeting-index(TI)of GDNL in liver.RESULTS:The prepared GDNL had an entrapment efficiency of 56.29%,particle diameter of(210?20)nm and TI of 5.213 in liver.CONCLUSION:The prepared GDNL has high entrapment efficiency and remarkable liver-targeting property.
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Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.
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Objective:To prepare lactosaminated matrine liposomes and to study its targetability to liver in mice and efficacy of anti-tumor activities on human hepatoma cell line HepG2 in vitro.Methods:The matrine liposomes were prepared using the reverse-phase evaporation-ultrasonic technique.Lactosylphosphatidy-lethanolamine(Lac-PE) was synthesized and used for modified matrine liposomes..the diameter and entrapment efficiency of it were determined.The RP-HPLC was used for the determination of matrine concentration in mice tissue.The cytotoxic effect of LML on human hepatoma cell line HepG2 in vitro was detected by thiazolyl blue(MTT)assay.Results:The LML were nice and uniform,the particle diameter was between 80 and 150 nm and envelopment rats was 48.1%.Compared with matrine solution,ML and LML exhibited long circulation time.Tissue distribution results proved that the area under curve of liver was significantly difference among modified matrine liposomes,regular matrine liposomes and matrine solutions(P
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The aim of the present study was to investigate the inhibitory effect of galactosides modified lamivudine (LA) on hepatitis B virus (HBV) and examine the liver targeting ability of lamivudine modified by galactosides in vitro and in vivo (mice). Lamivudine nanoparicles modified by galactosides (LAP GSLN) were prepared and delivered into 2.2.15 cells. After 10 days, hepatitis virus B e antigen (HBeAg) expression in 2.2.15 cells was detected by ELISA, and immune fluorescence levels of HBV DNA in the medium were examined by quantitative polymerase chain reaction (PCR). The cytotoxicity of LAP GSLN on 2.2.15 cells was observed as well. In the in vivo experiment, ten male mice were randomly divided into 2 groups: lap GSLN group (i.v.injection of LAP GSLN) and LA group (i.v.injection of LA). Lamivudine levels in serum, hepatic, renal, pulmonary, and splenic tissues were detected by reversed phase high performance liquid chromatography (RP HPLC). On the 6th day, the expression of HBeAg was found inhibited by LPA GSLN. HBV DNA replication was also inhibited by LAP GSLN on the 4th day. Hepatic LAP GSLN concentrations in LAP GSLN group were 3.3 fold higher that of the LA group. The above results suggested galactosides modified lamivudine could effectively inhibit the antigen expression and DNA replication of HBV, and it showed a high liver targeting ability in vivo .