Novel pancortin-Pablo protein interactions and methods of use thereof
AssigneeWyeth
InventorsRobert John Mark, Andrew Timothy Wood, Kamalakar Gulukota
Life of this patent
Estimating the term…
Abstract
This invention relates to newly identified human pancortin polypeptides, the interaction of the pancortin polypeptides with a Pablo polypeptide, the use of such polypeptides, as well as the production of such polypeptides. The invention relates also to identifying compounds which modulate the activity of a pancortin polypeptides and/or the interaction of a pancortin-Pablo polypeptide interaction, wherein modulators can be agonists, antagonists and/or inhibitors of pancortin and/or the pancortin/Pablo interaction and therefore potentially useful in therapy.
Claims97
An isolated polynucleotide encoding a human pancortin polypeptide, wherein the polynucleotide comprises a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:1, a degenerate variant thereof or a complement thereof.
An isolated polynucleotide encoding a human pancortin polypeptide, wherein the polynucleotide comprises a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:3, a degenerate variant thereof or a complement thereof.
An isolated polynucleotide encoding a human pancortin polypeptide, wherein the polynucleotide comprises a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:5, a degenerate variant thereof or a complement thereof.
An isolated polynucleotide encoding a human pancortin polypeptide, wherein the polynucleotide comprises the nucleotide sequence of SEQ ID NO:7 a degenerate variant thereof or a complement thereof.
An isolated polynucleotide which hybridizes with a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 1, SEQ ID NO:
- 3, SEQ ID NO:
- 5, SEQ ID NO:
- 7 or a complement thereof, under high stringency hybridization conditions.
An isolated human pancortin polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:1, a degenerate variant thereof or a complement thereof.
An isolated human pancortin polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:3, a degenerate variant thereof or a complement thereof.
An isolated human pancortin polypeptide encoded by a polynucleotide comprising a nucleotide sequence having at least 95% identity to the nucleotide sequence of SEQ ID NO:5, a degenerate variant thereof or a complement thereof.
An isolated human pancortin polypeptide encoded by a polynucleotide comprising a nucleotide sequence of SEQ ID NO:7, a degenerate variant thereof or a complement thereof.
An isolated human pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:2, a variant thereof or a fragment thereof.
An isolated human pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:4, a variant thereof or a fragment thereof.
An isolated human pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:6, a variant thereof or a fragment thereof.
An isolated human pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:
- 8, a variant thereof or a fragment thereof.
An antibody specific for a pancortin polypeptide comprising the amino acid sequence of SEQ ID NO:
- 2, SEQ ID NO:
- 4, SEQ ID NO:
- 6, SEQ ID NO:
- 8, a variant thereof or a fragment thereof.
An antibody specific for a pablo-pancortin polypeptide dimer.
An expression vector comprising a polynucleotide comprising the nucleotide sequence of SEQ ID NO:
- 1, a degenerate variant thereof, a complement thereof or a fragment thereof.
An expression vector comprising a polynucleotide comprising the nucleotide sequence of SEQ ID NO:3, a degenerate variant thereof, a complement thereof or a fragment thereof.
An expression vector comprising a polynucleotide comprising the nucleotide sequence of SEQ ID NO:5, a degenerate variant thereof, a complement thereof or a fragment thereof.
An recombinant expression vector comprising a polynucleotide comprising the nucleotide sequence of SEQ ID NO:7, a degenerate variant thereof, a complement thereof or a fragment thereof.
A neural cell line stably expressing a pancortin polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, a variant thereof or a fragment thereof.
A method for modulating apoptosis in a cell comprising modulating the activity of a pancortin polypeptide.
A method for modulating apoptosis in a cell comprising modulating the expression of a polynucleotide encoding a pancortin polypeptide.
A method for treating a subject for a nervous system disorder comprising modulating the activity of a pancortin polypeptide and/or modulating the expression of a polynucleotide encoding a pancortin polypeptide.
A method for assaying the effects of test compounds on the activity of a pancortin polypeptide comprising the steps of:
- (a) providing a transgenic animal comprising a polynucleotide encoding a pancortin polypeptide;
- (b) administering a test compound to the animal;
- and
- (c) determining the effects of the test compound on the activity of the pancortin in the presence and absence of the test compound.
A method for assaying the effects of test compounds on an animal with a genome comprising a functional disruption of a polynucleotide encoding a pancortin polypeptide, the method comprising:
- (a) providing a transgenic animal whose genome comprises a disruption of the endogenous polynucleotide encoding a pancortin polypeptide;
- (b) administering a test compound to the animal;
- and
- (c) determining the effects of the test compound on the activity of the pancortin polypeptide in the presence and absence of the test compound.
A method for assaying the effects of test compounds on the activity of a pancortin polypeptide comprising the steps of:
- (a) providing recombinant cells comprising a polynucleotide expressing a pancortin polypeptide;
- (b) contacting the cells with a test compound;
- and
- (c) determining the effects of the test compound on the activity of the pancortin in the presence and absence of the test compound.
A method for assaying the effects of test compounds on the binding interaction of pancortin and pablo polypeptides comprising the steps of:
- (a) providing yeast cells for a yeast two-hybrid system comprising a pancortin polypeptide and a pablo polypeptide;
- (b) contacting the cells with a test compound;
- and
- (c) determining the effect of the test compound on the binding interaction of the pancortin and pablo polypeptides in the presence and absence of the test compound.
A method of producing a pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, a variant thereof or a fragment thereof, comprising:
- (a) transfecting, transforming or infecting a recombinant host cell with an expression vector comprising a polynucleotide comprising a nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, or a degenerate variant thereof;
- (b) culturing the host cell under conditions sufficient for the production of the polypeptide;
- and
- (c) isolating the polypeptide from the culture.
A method for the treatment of a subject in need of reduced pancortin activity comprising:
- (a) administering to the subject a therapeutically effective amount of a pancortin antagonist;
- and/or
- (b) administering to the subject a polynucleotide encoding an antisense RNA polynucleotide comprising a nucleotide sequence that is a complement to a nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, a degenerate variant thereof or a fragment thereof.
A method for the diagnosis of a disease or the susceptibility to a disease in a subject related to the expression or activity of a pancortin polypeptide in the subject comprising:
- (a) determining the presence or absence of a mutation in a polynucleotide encoding a pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, or a fragment thereof;
- and/or
- (b) assaying for the presence of pancortin expression in a sample derived from the subject, wherein the pancortin expressed is a polynucleotide encoding a pancortin polypeptide comprising an amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, or a fragment thereof.
A composition for treating a hyperproliferative disease comprising a pancortin polypeptide comprising the amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, or SEQ ID NO:8 and a pablo polypeptide comprising the amino acid sequence of SEQ ID NO:10.
A nucleic acid molecule which is antisense to a pancortin mRNA molecule.
A method of inhibiting expression of a pancortin gene in a cell comprising providing said cell with an antisense nucleic acid.
A non-human transgenic mammal whose genome comprises an exogenous polynucleotide which encodes a pancortin polypeptide or a fragment thereof, wherein the polynucleotide expression is under the control of a regulated promoter.
78 The mammal of claim 77, wherein the polypeptide comprises an amino acid sequence of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6 or SEQ ID NO:8.
A non-human transgenic mammal whose genome comprises a homozygous disruption in its endogenous pancortin gene, wherein the disruption prevents the expression of a functional pancortin polypeptide.
A method for producing a non-human transgenic mammal whose genome comprises an exogenous polynucleotide which encodes a pancortin polypeptide or a fragment thereof comprising the steps of:
- (a) introducing into the pronucleus of a fertilized oocyte a polynucleotide comprising a nucleic acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5 or SEQ ID NO:7, wherein the polynucleotide is operatively linked to a promoter;
- (b) implanting the oocyte into a pseudopregnant non-human mammal, wherein the oocyte develops into an embryo;
- and
- (c) allowing the embryo to develop into a viable transgenic mammal.
A method for producing a non-human transgenic mammal whose genome comprises a disruption in its endogenous pancortin gene, the method comprising:
- (a) providing a polynucleotide encoding a pancortin polypeptide having a functional disruption;
- (b) introducing the disrupted polynucleotide into embryonic stem cells;
- (c) selecting those embryonic stem cells that comprise the disrupted polynucleotide;
- (d) introducing an embryonic stem cell of step (c) into a blastocyst;
- (e) transferring the blastocyst of step (d) to a pseudopregnant animal;
- and
- (f) allowing the transferred blastocyst to develop into a mammal chimeric for the disruption;
- wherein the disruption prevents the expression of a functional pancortin polypeptide.
Dependencies are inferred from the claim text.