Preparation of conjugates of oligodeoxynucleotides and lipid structures and their interaction with low-density lipoprotein

被引:24
作者
Rump, ET [1 ]
de Vrueh, RLA [1 ]
Sliedregt, LAJM [1 ]
Biessen, EAL [1 ]
van Berkel, TJC [1 ]
Bijsterbosch, MK [1 ]
机构
[1] Leiden Amsterdam Ctr Drug Res, Div Biopharmaceut, NL-2300 RA Leiden, Netherlands
关键词
D O I
10.1021/bc970176z
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The high expression level of receptors for low-density lipoprotein (LDL) on tumor cells makes LDL an attractive carrier for selective delivery of drugs to these cells. The aim of this study is to allow incorporation of oncogene-directed antisense oligodeoxynucleotides (ODNs) into the lipid moiety of LDL. Therefore, ODNs were conjugated with oleic acid, cholesterol, and several other steroid lipids. These latter steroid lipids were synthesized starting from bile acids and were varied in lipophilicity by attaching oleic acid ester structures. The lipid structures, activated as pentafluorophenyl esters, were conjugated in solution phase to 3'-amino-tailed ODNs. The ODNs conjugated with lithocholic acid, oleic acid, and cholesterol could easily be purified by reversed phase (RP)-HPLC. However, the ODNs conjugated with the oleoyl steroid ester structures irreversibly bound to the column material. These highly lipidic ODNs were separated from the nonconjugated ODN by electrophoresis in a 1% low-melting agarose gel containing 0.1% Tween 20. This method was found to be very effective in isolating the ODNs conjugated to the oleoyl steroid ester structures. The ODNs conjugated with cholesterol and the oleoyl esters of lithocholic and cholenic acid associated readily and nearly completely with LDL. However, the less lipidic ODNs and the ODN conjugated with the dioleoyl ester of chenodeoxycholic acid did not and did incompletely associate, respectively. Lithocholic acid and oleic acid are probably not sufficiently lipophilic to induce association with LDL, whereas the dioleoyl ester structure is probably too bulky and extended to allow partitioning into the lipid moiety of LDL. We conclude that several of the lipid-ODNs can associate readily with LDL, enabling delivery of oncogene-directed antisense ODNs via the LDL receptor pathway.
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页码:341 / 349
页数:9
相关论文
共 44 条
[1]  
Alahari SK, 1996, MOL PHARMACOL, V50, P808
[2]   BATHOPHENANTHROLINE-RUTHENIUM(II) COMPLEXES AS NON-RADIOACTIVE LABELS FOR OLIGONUCLEOTIDES WHICH CAN BE MEASURED BY TIME-RESOLVED FLUORESCENCE TECHNIQUES [J].
BANNWARTH, W ;
SCHMIDT, D ;
STALLARD, RL ;
HORNUNG, C ;
KNORR, R ;
MULLER, F .
HELVETICA CHIMICA ACTA, 1988, 71 (08) :2085-2099
[3]   SYNTHESIS OF ALKYLATING OLIGONUCLEOTIDE DERIVATIVES CONTAINING CHOLESTEROL OR PHENAZINIUM RESIDUES AT THEIR 3'-TERMINUS AND THEIR INTERACTION WITH DNA WITHIN MAMMALIAN-CELLS [J].
BOUTORIN, AS ;
GUSKOVA, LV ;
IVANOVA, EM ;
KOBETZ, ND ;
ZARYTOVA, VF ;
RYTE, AS ;
YURCHENKO, LV ;
VLASSOV, VV .
FEBS LETTERS, 1989, 254 (1-2) :129-132
[4]   Rapid Routes of Synthesis of Chemically Reactive and Highly Radioactively Labeled alpha- and beta-Oligonucleotide Derivatives for in Vivo Studies [J].
Boutorine, A. S. ;
Le Doan, T. ;
Battioni, J. P. ;
Mansuy, D. ;
Dupre, D. ;
Helene, Claude .
BIOCONJUGATE CHEMISTRY, 1990, 1 (05) :350-356
[5]   REVERSIBLE COVALENT ATTACHMENT OF CHOLESTEROL TO OLIGODEOXYRIBONUCLEOTIDES FOR STUDIES OF THE MECHANISMS OF THEIR PENETRATION INTO EUKARYOTIC CELLS [J].
BOUTORINE, AS ;
KOSTINA, EV .
BIOCHIMIE, 1993, 75 (1-2) :35-41
[6]   Chlorin-oligonucleotide conjugates: Synthesis, properties, and red light-induced photochemical sequence-specific DNA cleavage in duplexes and triplexes [J].
Boutorine, AS ;
Brault, D ;
Takasugi, M ;
Delgado, O ;
Helene, C .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1996, 118 (40) :9469-9476
[7]  
BOUTORINE AS, 1994, ANGEW CHEM INT EDIT, V33, P2462
[8]   A RECEPTOR-MEDIATED PATHWAY FOR CHOLESTEROL HOMEOSTASIS [J].
BROWN, MS ;
GOLDSTEIN, JL .
SCIENCE, 1986, 232 (4746) :34-47
[9]  
CHEN SH, 1986, J BIOL CHEM, V261, P2918
[10]   CHOLESTERYL-CONJUGATED PHOSPHOROTHIOATE OLIGODEOXYNUCLEOTIDES MODULATE CYP2B1 EXPRESSION IN-VIVO [J].
DESJARDINS, J ;
MATA, J ;
BROWN, T ;
GRAHAM, D ;
ZON, G ;
IVERSEN, P .
JOURNAL OF DRUG TARGETING, 1995, 2 (06) :477-485