Delivery of Antisense Oligonucleotides Using HPMA Polymer:  Synthesis of A Thiol Polymer and Its Conjugation to Water-Soluble Molecules

Wang, Laixin; Kristensen, Jakob; Ruffner, Duane E.

doi:10.1021/bc980034k

Cited by 79 publications

(78 citation statements)

References 39 publications

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“…Disulfide linkage between TFO and cholesterol is expected to be cleaved inside the cells by disulfide exchange with intracellular thiols or by the action of redox enzyme (Feener et al, 1990;Oberhauser and Wagner, 1992;Wang et al, 1998). Therefore, TFO-Chol was treated with DTT and incubated with the target duplex DNA to form triplex at physiological conditions and then was determined using gel shift assay.…”

Section: Resultsmentioning

confidence: 99%

Enhanced Hepatic Uptake and Bioactivity of Type α1(I) Collagen Gene Promoter-Specific Triplex-Forming Oligonucleotides after Conjugation with Cholesterol

Cheng

Guntaka

et al. 2006

J Pharmacol Exp Ther

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A triplex-forming oligonucleotide (TFO) specific for type ␣1(I) collagen promoter is a promising candidate for treating liver fibrosis. Earlier, we determined the pharmacokinetics and biodistribution of TFO after systemic administration into normal and fibrotic rats. In this study, we conjugated cholesterol to the 3Ј end of the TFO via a disulfide bond and determined its cellular and nuclear uptake and bioactivity using HSC-T6 cell lines in vitro, followed by biodistribution at whole-body, organ (liver), and subcellular levels. Conjugation with cholesterol had little effect on the triplex-forming ability of the TFO with target duplex DNA, and the cellular uptake of 33 P-TFO-cholesterol (Chol) increased by 2-to approximately 4-fold. Real-time reverse transcriptase-polymerase chain reaction analysis after transfection of HSC-T6 cells with TFO-Chol or TFO indicated that TFO-Chol had higher inhibition on type ␣1(I) collagen primary transcript than naked TFO at low concentration (200 nM) but showed similar inhibition at higher concentration (500 and 1000 nM). There was increase in the inhibition on primary transcript with transfection time. The hepatic uptake of 33 P-TFO-Chol after systemic administration was 72.22% of the dose compared with 45.8% of 33 P-TFO. There was significant increase in the uptake of 33 P-TFO-Chol by hepatic stellate cells and hepatocytes. More importantly, the nuclear uptake of TFOChol was higher than TFO in cell culture system and in vivo studies. In conclusion, TFO-Chol is a potential antifibrotic agent.

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Section: Resultsmentioning

confidence: 99%

Enhanced Hepatic Uptake and Bioactivity of Type α1(I) Collagen Gene Promoter-Specific Triplex-Forming Oligonucleotides after Conjugation with Cholesterol

Cheng

Guntaka

et al. 2006

J Pharmacol Exp Ther

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“…For this purpose, various pyridyldisulfide-functionalized acrylates and methacrylates have been developed that can be polymerized using conventional free-radical polymerization as well as ATRP (Table 1) and that are stable towards hydrolysis and aminolysis below pH 8. [50] A particular advantage of the pyridyldisulfide group is that thiol exchange generates 2-pyridinethione, which is an inert leaving group and has a characteristic UV/Vis absorbance spectrum that is distinctly different from that of the pyridyldisulfide functionality. [50] Terpolymers (copolymers made from three different monomers) containing pyridyldisulfide propylacrylate units have been modified with peptides bearing a free cysteine group to 86 and 35 % (relative to pyridyldisulfide units) at pH 6 and 10, respectively.…”

Section: Modification Of Polymers By Thiol Exchangementioning

confidence: 99%

Synthesis of Functional Polymers by Post‐Polymerization Modification

2008

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Post-polymerization modification is based on the direct polymerization or copolymerization of monomers bearing chemoselective handles that are inert towards the polymerization conditions but can be quantitatively converted in a subsequent step into a broad range of other functional groups. The success of this method is based on the excellent conversions achievable under mild conditions, the excellent functional-group tolerance, and the orthogonality of the post-polymerization modification reactions. This Review surveys different classes of reactive polymer precursors bearing chemoselective handles and discusses issues related to the preparation of these reactive polymers by direct polymerization of appropriately functionalized monomers as well as the post-polymerization modification of these precursors into functional polymers.

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“…These polymers have been studied and used to deliver many types of drugs. The main areas of research include the delivery of anticancer drugs (3), site-specific delivery in the gastrointestinal tract (16), antisense therapy (17,18), hydrogels (19,20), and modified surfaces as platforms for epitope display (21).…”

mentioning

confidence: 99%

The cytoplasmic escape and nuclear accumulation of endocytosed and microinjected HPMA copolymers and a basic kinetic study in hep G2 cells

et al. 2001

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The development of macromolecules as drugs and drug carriers requires knowledge of their fate in cells. To this end, we studied the internalization and subcellular fate of N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers in Hep G2 (human hepatocellular carcinoma) cells. Semiquantitative fluorometry confirmed that galactose was an effective ligand for receptormediated endocytosis for Hep G2 cells. The rate of internalization of a galactose-targeted copolymer was almost 2 orders of magnitude larger than that of the nontargeted copolymer. Confocal fluorescent microscopy of both fixed and live cells revealed that the polymer entered the cells by endocytosis. After longer incubation times (typically >8 hours), polymer escaped from small vesicles and distributed throughout the cytoplasm and nuclei of the cells. Polymer that entered the cytoplasm tended to accumulate in the nucleus. Microinjection of the HPMA copolymers into cells' cytoplasm and nuclei indicated that the polymers partitioned to the nucleus. The data from fixed cells was confirmed by microscopy of live, viable cells. To examine the effect of the fluorescent dye on the intracellular fate, polymers with fluorescein, Oregon Green 488, Lissamine rhodamine B, and doxorubicin were tested; no significant differences were observed.

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Delivery of Antisense Oligonucleotides Using HPMA Polymer: Synthesis of A Thiol Polymer and Its Conjugation to Water-Soluble Molecules

Cited by 79 publications

References 39 publications

Enhanced Hepatic Uptake and Bioactivity of Type α1(I) Collagen Gene Promoter-Specific Triplex-Forming Oligonucleotides after Conjugation with Cholesterol

Enhanced Hepatic Uptake and Bioactivity of Type α1(I) Collagen Gene Promoter-Specific Triplex-Forming Oligonucleotides after Conjugation with Cholesterol

Synthesis of Functional Polymers by Post‐Polymerization Modification

The cytoplasmic escape and nuclear accumulation of endocytosed and microinjected HPMA copolymers and a basic kinetic study in hep G2 cells

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