Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Method of regulating cell metabolism or physiology
Reexamination Certificate
2001-09-28
2004-02-17
Lacourciere, Karen (Department: 1635)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Method of regulating cell metabolism or physiology
C435S325000, C435S006120, C435S091100, C536S024300, C536S024310, C536S024330, C536S063000
Reexamination Certificate
active
06692960
ABSTRACT:
FIELD OF THE INVENTION
The present invention provides compositions and methods for modulating the expression of sphingosine-1-phosphate lyase. In particular, this invention relates to compounds, particularly oligonucleotides, specifically hybridizable with nucleic acids encoding sphingosine-1-phosphate lyase. Such compounds have been shown to modulate the expression of sphingosine-1-phosphate lyase.
BACKGROUND OF THE INVENTION
Sphingolipids are highly enriched in the membranes of most mammalian cells where they were originally thought to play a predominantly structural role as components of the lipid bilayer. It is now appreciated however, that sphingolipid metabolism is a dynamic process and that metabolites of sphingomyelin function as second messengers in cell growth, survival, and death. The dynamic balance between the concentrations of these sphingolipid metabolites and the regulation of opposing signaling pathways, is an important factor that determines whether a cell survives or dies.
Sphingosine-1-phosphate is a phosphorylated derivative of sphingosine, the structural backbone of all sphingolipids, and was initially described as an intermediate in the degradation of long-chain sphingoid bases. However, the discovery that sphingosine-1-phosphate is a potent mitogen for diverse cell types suggested that this metabolite might play other important physiological roles. Sphingosine-1-phosphate has been implicated in a wide variety of biological processes, including mobilization of intracellular calcium, regulation of cytoskeletal organization, as well as cell growth, differentiation, survival, and motility (Spiegel and Milstien,
Biochim. Biophys. Acta,
2000, 1484, 107-116). Attempts to characterize the mechanisms through which sphingosine-1-phosphate exerts such a broad array of actions have revealed that this sphingolipid metabolite is a member of a new class of lipid second messengers that has both intracellular and extracellular actions (Spiegel and Milstien,
Biochim. Biophys. Acta,
2000, 1484, 107-116). Since sphingosine-1-phosphate acts as an intracellular messenger, its synthesis should be stimulation-dependent and transient (Yatomi et al.,
Prostaglandins,
2001, 64, 107-122).
Sphingosine-1-phosphate lyase (also known as SPL, SGPL1 and KIAA1252) cleaves sphingosine-1-phosphate into fatty aldehyde and phosphoethanolamine and thus, plays a role in keeping intracellular levels of sphingosine-1-phosphate at low levels (Yatomi et al.,
Prostaglandins,
2001, 64, 107-122). The gene for this enzyme was initially identified in yeast (Saba et al.,
J. Biol. Chem.,
1997, 272, 26087-26090) and the human gene was later cloned and mapped to chromosome 10q20 (Van Veldhoven et al.,
Biochim. Biophys. Acta,
2000, 1487, 128-134). Three different sphingosine-1-phosphate lyase mRNAs of approximately 4.0, 5.8 and 6.7 kb are observed with their highest levels seen in liver and kidney (Van Veldhoven et al.,
Biochim. Biophys. Acta,
2000, 1487, 128-134).
Disclosed and claimed in U.S. Pat. No. 5,187,562 and corresponding PCT publication WO 99/38983 are isolated polynucleotides encoding human sphingosine-1-phosphate lyase and polynucleotides fully complementary to said polynucleotides encoding human sphingosine-1-phosphate lyase (Duckworth et al., 1999; Duckworth et al., 1999).
Disclosed and claimed in PCT publication WO 99/16888 are polynucleotide sequences encoding sphingosine-1-phosphate lyase polypeptides, isolated polynucleotides comprising at least 100 nucleotides complementary to said sphingosine-1-phosphate lyase polynucleotides, pharmaceutical compositions of polynucleotides or antibodies which inhibit the expression of an endogenous sphingosine-1-phosphate lyase gene, and a transgenic animal in which sphingosine-1-phosphatase activity is reduced compared to a wild-type animal (Saba and Zhou, 1999).
Since sphingosine-1-phosphate has been implicated in cell signaling, impaired degradation of this lipid might have severe consequences during neonatal development or even be fatal (Van Veldhoven et al.,
Biochim. Biophys. Acta,
2000, 1487, 128-134).
Sphingosine-1-phosphate has been recently implicated in resistance to the anticancer drug cisplatin and suggestions have been made that manipulating the levels of sphingosine-1-phosphate could be an important therapeutic avenue by potentiating tumor cells to be more sensitive to cisplatin or other drugs (Li et al.,
Microbiology,
2000, 146, 2219-2227).
The synthesis of a small molecule inhibitor of sphingosine-1-phosphate lyase, 2-vinyldihydrosphingosine-1-phosphate, has been reported by Boumendjel et al. (Boumendjel and Miller,
Tetrahedron Lett.,
1994, 35, 819-822).
Currently, there are no known therapeutic agents that effectively inhibit the synthesis of sphingosine-1-phosphate lyase. To date, investigative strategies aimed at modulating sphingosine-1-phosphate lyase function have involved the use of antibodies and small molecule inhibitors. Consequently, there remains a long felt need for additional agents capable of effectively inhibiting sphingosine-1-phosphate lyase function.
Antisense technology is emerging as an effective means for reducing the expression of specific gene products and may therefore prove to be uniquely useful in a number of therapeutic diagnostic and research applications for the modulation of sphingosine-1-phosphate lyase expression.
The present invention provides compositions and methods for modulating sphingosine-1-phosphate lyase expression, including modulation of alternate mRNA transcripts of sphingosine-1-phosphate lyase.
SUMMARY OF THE INVENTION
The present invention is directed to compounds, particularly antisense oligonucleotides, which are targeted to a nucleic acid encoding sphingosine-1-phosphate lyase, and which modulate the expression of sphingosine-1-phosphate lyase. Pharmaceutical and other compositions comprising the compounds of the invention are also provided. Further provided are methods of modulating the expression of sphingosine-1-phosphate lyase in cells or tissues comprising contacting said cells or tissues with one or more of the antisense compounds or compositions of the invention. Further provided are methods of treating an animal, particularly a human, suspected of having or being prone to a disease or condition associated with expression of sphingosine-1-phosphate lyase by administering a therapeutically or prophylactically effective amount of one or more of the antisense compounds or compositions of the invention.
DETAILED DESCRIPTION OF THE INVENTION
The present invention employs oligomeric compounds, particularly antisense oligonucleotides, for use in modulating the function of nucleic acid molecules encoding sphingosine-1-phosphate lyase, ultimately modulating the amount of sphingosine-1-phosphate lyase produced. This is accomplished by providing antisense compounds which specifically hybridize with one or more nucleic acids encoding sphingosine-1-phosphate lyase. As used herein, the terms “target nucleic acid” and “nucleic acid encoding sphingosine-1-phosphate lyase” encompass DNA encoding sphingosine-1-phosphate lyase, RNA (including pre-mRNA and mRNA) transcribed from such DNA, and also cDNA derived from such RNA. The specific hybridization of an oligomeric compound with its target nucleic acid interferes with the normal function of the nucleic acid. This modulation of function of a target nucleic acid by compounds which specifically hybridize to it is generally referred to as “antisense”. The functions of DNA to be interfered with include replication and transcription. The functions of RNA to be interfered with include all vital functions such as, for example, translocation of the RNA to the site of protein translation, translation of protein from the RNA, splicing of the RNA to yield one or more mRNA species, and catalytic activity which may be engaged in or facilitated by the RNA. The overall effect of such interference with target nucleic acid function is modulation of the expression of sphingosine-1-phosphate lyase. In the context of the present invention, “modulati
Bennett C. Frank
Freier Susan M.
Isis Pharmaceuticals , Inc.
Lacourciere Karen
Licata & Tyrrell P.C.
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