Figures & data
Figure 1. Doxorubicin content leakage profile from ScApt-Doxil and 5TR1-Doxil compared to Doxil® at 37 °C at the presence of 30% FCS. Diluted liposomes were dialyzed at 37 °C in 100 mL of the media. At various time points, the percent of doxorubicin retained encapsulated was measured (n = 3).
Table 1. Properties of liposomal preparations. Each value represents mean ± SD (n = 3).
Figure 2. Confirmation of liposome–aptamer conjugation. Samples are loaded onto the agarose gel. After running time, the gel was visualized by UV light. Lane 1: free 5TR1 aptamer; 2: 5TR1-liposomes; 3: liposome without conjugation.
Table 2. In vitro cytotoxicity effect (IC50) of liposomal preparations and free DOX against C26 and CHO-K1 cells after 72 h exposure times. Data represented as mean ± SD (n = 3).
Figure 3. Flow cytometry analysis of doxorubicin cell uptake. C26 cell line (A), CHO-K1 cell line (B) and pre-treated C26 cells with anti-MUC1 antibody (C) were incubated with Doxil and 5TR1-Doxil at 37 °C in the presence of 10% FCS. After 30-min, uptake of doxorubicin was assessed by flow cytometry. Results, expressed as mean fluorescence intensity (MFI), represented mean ± SEM of three independent experiments. There is a significant difference (p < .1) of Doxil compared with 5TR1-Doxil in cellular uptake by C26 cell line. However, cellular uptake by CHO-K1 cells is not affected by 5TR1 targeting. Cellular uptake by C26 cell line significantly decreases after treatment of cell with anti-MUC1 antibody (p < .01).
Figure 4. Liposome–cell association at different time points. C26 cells were treated with different preparations at a concentration of 100 nmol phospholipid/mL for 1, 3 and 6 h of incubation at either 37 °C or 4 °C. Then, the cells were detached, washed and lysed in acidified isopropanol and percent of doxorubicin associated with cells was measured based on the amount of doxorubicin assayed in the medium and amount of doxorubicin detected in cell lysate. Data are expressed as mean ± SD (n = 3).
Table 3. Pharmacokinetic parameters using non-compartment model in mice following singel intravenous injection of 15 mg/kg of Doxil-mimic, ScApt-PL-DOX and 5TR1-PL-DOX.
Table 4. AUC (µh) (24–48 h) of various tissues in mice-bearing C26 carcinoma after injection of 15 mg/kg doxorubicin as Doxil, ScApt-Doxil and 5TR1-Doxil.
Figure 6. Therapeutic efficacy of various liposomal preparations in female BALB/c mice after i.v. administration of a single dose of 15 mg/kg liposomal doxorubicin or dextrose 5% on day 8 after tumor inoculation. (A) Percentage change in animal body weight, (B) tumour growth rate, (C) survival curve and (D) survival curve with error bars. Data represented as mean ± SE (n = 5).
Table 5. Therapeutic efficacy data of liposomal preparations in mice-bearing C26 colon carcinoma.
Table 6. Number of at risk in every group at days 50 and 60 post-injections.