Of antisense oligonucleotide

A newer, highly experimental approach to anxiety therapy is the use of antisense oligonucleotides to the anxiogenic peptide, NPY (44). 

Antisense Oligonucleotides. Figure 1 Schematic representation of the action of antisense oligonucleotides. They bind to their respective target mRNA preventing protein translation. 

CpG stands for cytosine phosphate guanine dinucleotide in a particular sequence context. CpG motifs are responsible for proliferative effects of antisense oligonucleotides, particularly with respect to B-lymphocytes. Die optimal immune-stimulatory consensus sequence surrounding CpG is R1R2CGY1Y2, where R1 is a purine (mild preference for G), R2 is a purine or T (preference for A), and Y1 and Y2 are pyrimidines (preference for T). 

A Bochot, P Couvreur, E Fattal. Intravitreal administration of antisense oligonucleotides potential of liposomal delivery. Prog Retin Eye Res 19(2) 131—147, 2000. 

Crooke, R.M., In vitro toxicology and pharmacokinetics of antisense oligonucleotides, Anti-Cancer Drug Design, 1991, 6, 609-646. 

Yessine MA, Meier C, Petereit HU et al (2006) On the role of methacrylic acid copolymers in the intracellular delivery of antisense oligonucleotides. Eur J Pharm Biopharm 63 1-10... 

Yoo, H., and Juliano, R.L. (2000) Enhanced delivery of antisense oligonucleotides with fluorophore-con-jugated PAMAM dendrimers. Nucleic Acids Res. 28, 4225-4231. 

Orr, R. and O Neill, C. 2000. Patent review therapeutic applications of antisense oligonucleotides, 1999-2000. 

An alternative system, which effectively results in the introduction of antisense oligonucleotides into the cell, entails application of gene therapy. In this case, a gene, which when transcribed yields (antisense) mRNA of appropriate nucleotide sequence, is introduced into the cell by a retroviral or other appropriate vector. This approach, as applied to the treatment of cancer and AIDS, is being appraised in a number of trials. 

Hughes, M.D., Hussain, M., Nawaz, Q., Sayyed, P., and Akhtar, S. 2001. The cellular delivery of antisense oligonucleotides and ribozymes. Drug Discovery Today 6(6), 303-315. 

Rubenstein, M., Tsui, P., and Guinan, P. 2004. A review of antisense oligonucleotides in the treatment of human disease. Drugs of the Future 29(9), 893-909. 

Bioaffinity chromatography, 6 399—400 Bioantimutagen, vanillin as, 25 556 Bioassay dyes, 9 518 Bioassays, microfluidics in, 26 968-969 Bioaugmentation, defined, 3 758t Bioaugmentation/bioremediation effluent treatment, 9 436, 438 Bioavailability, of antisense oligonucleotides, 17 628 Biocatalysis, 3 668-683 16 395. See also Biocatalyst entries... 

Jasti BR, Zhou S, Mehta RC, Li X (2000) Permeability of antisense oligonucleotides through porcine buccal mucosa. Int J Pharm 208 35-39... 

Table 1 Entrapment of Antisense Oligonucleotide in the Absence of Polyethyleneglycol-Ceramide ...

Semple SC, Klimuk SK, Harasym TO, et al. Efficient encapsulation of antisense oligonucleotides in lipid vesicles using ionizable aminolipids formation of novel small multilamellar vesicle structures. Biochim Biophys Acta 2001 1510 152. 

Gelfi, C., Perego, M., Morelli, S., Nicolin, A., and Righetti, P. G. (1996). Analysis of antisense oligonucleotides by capillary electrophoresis, gel-slab electrophoresis, and HPLC a comparison. 

Khan, K., Van Schepdael, A., Saison-Behmoaras, T., Van Aerschot, A., and Hoogmartens, J. (1998). Analysis of antisense oligonucleotides by on-capillary isotachophoresis and capillary polymer sieving electrophoresis. Electrophoresis 19, 2163-2168. 

Effenhauser, C. S., Paulus, A., Manz, A., and Widmer, H. M. (1994). High-speed separation of antisense oligonucleotides on a micromachined capillary electrophoresis device. Anal. Chem. 66, 2949-2953. 

Example 56 the Isis Pharmaceutical group in their extensive investigations of antisense oligonucleotides as therapeutics has described the synthesis of 3 -C-methylene nucleoside phosphonoamidites for the new backbone modification of oligonucleotides [90]. This paper gives good insight into tricoordinate phosphorus and related H-phosphonate chemistry in the service of nucleotide synthesis. 

Microspheres and nanoparticles often consist of biocompatible polymers and belong either to the soluble or the particle type carriers. Besides the aforementioned HPMA polymeric backbone, carriers have also been prepared using dextrans, ficoll, sepharose or poly-L-lysine as the main carrier body. More recently alginate nanoparticles have been described for the targeting of antisense oligonucleotides [28]. As with other polymeric carrier systems, the backbone can be modified with e.g. sugar molecules or antibody fragments to introduce cellular specificity. 

A selective method of preventing the expression of adhesion molecules or cytokines is the use of antisense oligonucleotides. These oligonucleotides are short sequences of nucleic acids complementary to mRNA sequences of specific proteins of interest. If delivered to the cytoplasmic compartment of cells these oligonucleotides are able to form a complex with their target mRNA. In this way the translation of mRNA into protein by ribosomes is inhibited. The subsequent mRNA degradation by RNAse H results in reduced expression of the protein (see also Chapter 5 for a description of antisense ohgonucleotides as therapeutic modalities). 

Several lines of evidence suggest that the locus coendeus is the site of the a2A-agonist-mediated sedative response. The a2A-subtype is abundantly expressed in locus coeruleus neurons (Wang et al. 1996). Administration of antisense oligonucleotides for the a2A-receptor subtype into the locus coeruleus of rats attenuated the hypnotic effect of dexmedetomidine reversibly (Mizobe et al. 1996). In the locus coeruleus of wild-type mice, 02-agonists suppressed the spontaneous firing rate of neurons but did not alter... 

FICURE 1-1. Intracerebral infusion of antisense oligonucleotides (AS-ODN) that were targeted to the cloned CRH, and CRH2 receptor mRNA prevent translation into the receptor protein. Only a knock-down of the CRH, receptor, but not of the CRH2 receptor, reduces anxiety-related behavior in rats that were exposed to the elevated plus-maze test after central corticotropin-releasing hormone (CRH) administration. P <. 05, P <. 01. 

Bongartz, J.P., Aubertin, A.M., Milhaud, P.G., Lebleu, B. (1994). Improved biological activity of antisense oligonucleotides conjugated to a fusogenic peptide. Nucleic Acids Res., 22, 4681 1688. 

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