Evaluation of intracellular trafficking and clearance from HeLa cells of doxorubicin-bound block copolymers

Sakai‐Kato, Kumiko; Ishikura, Keiko; Oshima, Yuki; Tada, Minoru; Suzuki, Tomoya; Ishii‐Watabe, Akiko; Yamaguchi, Teruhide; Nishiyama, Nobuhiro; Kataoka, Kazunori; Kawanishi, Toru; Okuda, Haruhiro

doi:10.1016/j.ijpharm.2011.12.022

Cited by 25 publications

(29 citation statements)

References 45 publications

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“…The conjugation was designed not to release doxorubin in vivo [12]. Indeed, we confirmed that the in vitro cytotoxicity of the doxorubicin-conjugated polymers was negligible [14].…”

Section: Introductionsupporting

confidence: 66%

“…Samples for HPLC analysis were prepared as described previously [14,25,26]. Tissue samples were weighed and homogenized on ice by using a hand homogenizer (MH-1000, As One, Osaka) and suspended in PBS at a concentration of 500 µL/100 mg tissue (liver, brain, lung, kidney, fat, muscle, small intestine, and feces) or 500 µL/50 mg tissue (heart and spleen), Plasma and urine samples were sonicated with an ultrasonic liquid processor (ASTRASON 3000, Misonix, NY) and treated as described previously [25].…”

Section: Sample Preparation For Hplc Analysismentioning

confidence: 99%

“…The amount of doxorubicin-conjugated polymer in the biological samples was determined by measuring the amount of doxorubicinone, which was released by HCl-treated acid hydrolysis of doxorubicin-conjugated polymer after sampling ( Supplementary Fig.1) [14,25,26]. Doxorubicin-conjugated polymer contained negligible amounts of free doxorubicinone (0.023% w/w) and free doxorubicin (0.017% w/w) in the injection sample to mice.…”

Section: Quantification Of Doxorubicin-conjugated Polymer In Biologicmentioning

confidence: 99%

“…When free doxorubicin, which is a well-known substrate of ABCB1 [14], was 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 18 administered intravenously to wild-type and Mdr1a/1b(-/-) mice, the plasma concentration of doxorubicin was significantly higher in Mdr1a/1b(-/-) mice compared with that in wild-type mice, from 3 h after administration (P < 0.01) (Fig. 2B).…”

Section: Plasma Concentration Of the Doxorubicin-conjugated Polymermentioning

confidence: 99%

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Effect of Knockout of Mdr1a and Mdr1b ABCB1 Genes on the Systemic Exposure of a Doxorubicin-Conjugated Block Copolymer in Mice

Sakai‐Kato

Nanjo

Kusuhara³

et al. 2015

Mol. Pharmaceutics

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We previously elucidated that ATP-binding cassette subfamily B member 1 (ABCB1) mediates the efflux of doxorubicin-conjugated block copolymers from HeLa cells. Here, we investigated the role of ABCB1 in the in vivo behavior of a doxorubicin-conjugated polymer in Mdr1a/1b(-/-) mice. The area under the curve for intravenously administered polymer in Mdr1a/1b(-/-) mice was 2.2-fold greater than that in wild-type mice. The polymer was mostly distributed in the liver followed by spleen and less so in the brain, heart, kidney, and lung. The amount of polymer excreted in the urine was significantly decreased in Mdr1a/1b(-/-) mice. The amounts of polymers excreted in the feces were similar in both groups despite the higher systemic exposure in Mdr1a/1b(-/-) mice. Confocal microscopy images showed polymer localized in CD68(+) macrophages in the liver. These results show that knockout of ABCB1 prolonged systemic exposure of the doxorubicin-conjugated polymer in mice. Our results suggest that ABCB1 mediated the excretion of doxorubicin-conjugated polymer in urine and feces. Our results provide valuable information about the behavior of block copolymers in vivo, which is important for evaluating the pharmacokinetics of active substances conjugated to block copolymers or the accumulation of block copolymers in vivo.

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“…The conjugation was designed not to release doxorubin in vivo [12]. Indeed, we confirmed that the in vitro cytotoxicity of the doxorubicin-conjugated polymers was negligible [14].…”

Section: Introductionsupporting

confidence: 66%

Section: Sample Preparation For Hplc Analysismentioning

confidence: 99%

Section: Quantification Of Doxorubicin-conjugated Polymer In Biologicmentioning

confidence: 99%

Section: Plasma Concentration Of the Doxorubicin-conjugated Polymermentioning

confidence: 99%

See 2 more Smart Citations

Effect of Knockout of Mdr1a and Mdr1b ABCB1 Genes on the Systemic Exposure of a Doxorubicin-Conjugated Block Copolymer in Mice

Sakai‐Kato

Nanjo

Kusuhara³

et al. 2015

Mol. Pharmaceutics

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“…35 Then free DOX, DNCHGC, or DCHGC micelles (equivalent DOX concentration: 5 µg/mL) in RPMI 1640 medium supplemented with 10% FBS were added to each designated dish. After 2 hours incubation, all media were removed.…”

Section: Confocal Microscope Studymentioning

confidence: 99%

Fabrication and characterization of nuclear localization signal-conjugated glycol chitosan micelles for improving the nuclear delivery of doxorubicin

Yu¹,

Xie²,

Zheng³

et al. 2012

IJN

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Background: Supramolecular micelles as drug-delivery vehicles are generally unable to enter the nucleus of nondividing cells. In the work reported here, nuclear localization signal (NLS)-modified polymeric micelles were studied with the aim of improving nuclear drug delivery. Methods: In this research, cholesterol-modified glycol chitosan (CHGC) was synthesized. NLSconjugated CHGC (NCHGC) was synthesized and characterized using proton nuclear magnetic resonance spectroscopy, dynamic light scattering, and fluorescence spectroscopy. Doxorubicin (DOX), an anticancer drug with an intracellular site of action in the nucleus, was chosen as a model drug. DOX-loaded micelles were prepared by an emulsion/solvent evaporation method. The cellular uptake of different DOX formulations was analyzed by flow cytometry and confocal laser scanning microscopy. The cytotoxicity of blank micelles, free DOX, and DOX-loaded micelles in vitro was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in HeLa and HepG2 cells. Results: The degree of substitution was 5.9 cholesterol and 3.8 NLS groups per 100 sugar residues of the NCHGC conjugate. The critical aggregation concentration of the NCHGC micelles in aqueous solution was 0.0209 mg/mL. The DOX-loaded NCHGC (DNCHGC) micelles were observed as being almost spherical in shape under transmission electron microscopy, and the size was determined as 248 nm by dynamic light scattering. The DOX-loading content of the DNCHGC micelles was 10.1%. The DOX-loaded micelles showed slow drug-release behavior within 72 hours in vitro. The DNCHGC micelles exhibited greater cellular uptake and higher amounts of DOX in the nuclei of HeLa cells than free DOX and DOX-loaded CHGC (DCHGC) micelles. The half maximal inhibitory concentration (IC 50 ) values of free DOX, DCHGC, and DNCHGC micelles against HepG2 cells were 4.063, 0.591, and 0.171 µg/mL, respectively. Moreover, the IC 50 values of free DOX (3.210 µg/mL) and the DCHGC micelles (1.413 µg/mL) against HeLa cells were nearly 6.96-and 3.07-fold (P , 0.01), respectively, higher than the IC 50 value of the DNCHGC micelles (0.461 µg/mL). Conclusion:The results of this study suggest that novel NCHGC micelles could be a potential carrier for nucleus-targeting delivery.

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Block Copolymer Micelles and Vesicles for Drug Delivery

Robertson¹,

Patikarnmonthon²,

Joseph³

et al. 2013

Engineering Polymer Systems for Improved Drug Delivery

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Evaluation of intracellular trafficking and clearance from HeLa cells of doxorubicin-bound block copolymers

Cited by 25 publications

References 45 publications

Effect of Knockout of Mdr1a and Mdr1b ABCB1 Genes on the Systemic Exposure of a Doxorubicin-Conjugated Block Copolymer in Mice

Effect of Knockout of Mdr1a and Mdr1b ABCB1 Genes on the Systemic Exposure of a Doxorubicin-Conjugated Block Copolymer in Mice

Fabrication and characterization of nuclear localization signal-conjugated glycol chitosan micelles for improving the nuclear delivery of doxorubicin

Block Copolymer Micelles and Vesicles for Drug Delivery

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