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Therapeutically useful triethyleneglycol cholesteryl oligonucleotides

A technology of cholestyl group and purpose, applied in the field of cholestyl group-coupled oligonucleotide composition, can solve the problems of toxicity, receptor weakening, immune sensitization and the like

Inactive Publication Date: 2005-03-23
BIONICHE LIFE SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many of these treatments are ineffective or toxic, have significant side effects, lead to the development of resistance or immune sensitization, and debilitate the recipient

Method used

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  • Therapeutically useful triethyleneglycol cholesteryl oligonucleotides
  • Therapeutically useful triethyleneglycol cholesteryl oligonucleotides
  • Therapeutically useful triethyleneglycol cholesteryl oligonucleotides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Nucleotide sequence preparation

[0071] Phosphodiester synthetic nucleotide sequences and coupled 3'-TEG cholestenyl synthetic nucleotide sequences were prepared by Sigma-Genosys (Woodlands, TX, USA) using Abacus segment synthesis technology. A sequence with a TEG cholestenyl group at its 3' end was synthesized on a TEG CPG support (Glen Research, Sterling, VA, USA). The sequence was dispersed in water or in dimethyl sulfoxide (DMSO) immediately before use.

Embodiment 2

[0073] cell

[0074] All cell lines were obtained from the American Type Culture Collection (ATCC, Rockyille, MD) and were grown in media recommended by ATCC. Breast cancer cell lines were cultured in media recommended by ATCC or described in published literature (Herrera-Gayol and Jothy, Int. J. Exp. Path., 82: 193, 2001). Table 1 shows the cell lines described in relevant literature, the source of the cell lines and their biopathological characteristics (Hackett et al., Cancer Inst., 58: 1795, 1977; Price et al., Cancer Res., 50: 717, 1990; Thompson et al., J. Cell Physiol., 150:534, 1992; and Peiper M, et al., Int. J. Cancer 71:993, 1997).

[0075] Table 1

[0076] cell line

illustrate

MEG-01*

Human chronic myeloid leukemia cell line

EL-4*

Murine T Lymphoma Cell Line

HIG-82**

Rabbit synovial cell line

MCF-7**

Human, breast adenocarcinoma (pleural effusion). Well differentiated. in vitro

and non-invasive ...

Embodiment 3

[0080] Apoptosis in MEG-01 cells was induced by SEQ ID NO:5 and SEQ ID NO:6.

[0081] Redistribution of plasma membrane phosphatidylserine is a characteristic of cells undergoing apoptosis (Martin et al., J. Exp. Med., 182:1545, 1995). The redistribution of phosphatidylserine in the plasma membrane during apoptosis was determined by flow cytometry using FITC (fluorescein isothiocyanate)-conjugated annexin V (BD Pharmingen, San Diego, CA). technically determined. The human chronic myeloid leukemia cell line MEG-01 cells that are positive for Philadelphia chromosome and have BCR-ABL gene fusion were used as 2.5×10 5 Concentration of cells / ml vs. final concentrations of 5.3, 26.5 and 53.0 μM of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 5, SEQ ID NO: 6 and cholestenyl-TEG phosphoramidite molecules Incubate together for 48 hours. The percentage of cells undergoing apoptosis following treatment with SEQ ID NOs: 1, 2, 5, 6, or cholestenyl-TEG phosphoramidites is reported in Table 2. ...

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PUM

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Abstract

The present invention provides a composition comprising a 5'-OH, 3'-TEG cholesteryl synthetic sequence wherein the sequence is SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8. The present invention provides methods of using this composition for induction of response in a cell, including but not limited to inhibition of cellular proliferation, induction of cell cycle arrest, induction of caspase activation, cleavage of poly(ADP-ribose) polymerase, induction of apoptosis or modulation of extracellular matrix-cell interactions, or combinations thereof, in cancer cells or synovial cells, and methods of using this composition for treating disease.

Description

field of invention [0001] The present invention relates to cholestenyl-conjugated oligonucleotide compositions and their use for inhibiting cell proliferation, inducing apoptosis, modifying cell cycle progression and modulating extracellular matrix-cell interactions. Background of the invention [0002] Proliferation is the culmination of cell development through the cell cycle, resulting in the division of one cell into two cells. The five major phases of the cell cycle are the G 0 , G 1 ,S,G 2 , and M period. in G 0 period, the cells are quiescent. Most cells in the body are in this phase at some point. in G 1 During the first phase, the cell divides according to the signal to produce the RNA and protein necessary for DNA synthesis. During S-phase (SE, early S-phase; SM, middle S-phase; and SL, late S-phase), cells replicate their DNA. in G 2 During this period, proteins are working in preparation for cell division. During mitosis (M) phase, a cell divides into ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61K47/48A61P19/02A61P35/00A61P35/02A61P43/00C07H21/00C12N15/117
CPCC12N2310/3515A61K47/4813C07H21/00A61K47/48215C12N15/117A61K47/48076C12N2310/18A61K47/60A61K47/547A61K47/555A61P19/02A61P35/00A61P35/02A61P43/00A61K47/50A61K31/7088C07H21/04
Inventor 安德烈亚·克里斯蒂娜·赫雷拉-盖奥尔N·C·菲利普斯M·C·菲利安
Owner BIONICHE LIFE SCI
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