NPC1L1 (NPC3) and methods of use thereof
AssigneeSchering Corporation
InventorsScott W. Altmann, Nicholas J. Murgolo, Luquan Wang, Michael P. Graziano
Life of this patent
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Abstract
The present invention provides rat and mouse NPC1L1 polypeptides and polynucleotides encoding the polypeptides. Also provided are methods for detecting agonists and antagonists of NPC1L1. Inhibitors of NPC1L1 can be used for inhibiting intestinal cholesterol absorption in a subject.
Claims44
An isolated polypeptide comprising 42 or more contiguous amino acids from an amino acid sequence selected from SEQ ID NOs:
- 2 and 12.
An isolated polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 2 and 12.
An isolated polynucleotide comprising a nucleotide sequence selected from SEQ ID NOs:
- 1 and 11.
An antibody which specifically binds to a polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 39-42.
A method for identifying an antagonist of NPC1L1 comprising:
- (a) contacting a host cell expressing a polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 2, 4 and 12 or a functional fragment thereof on a cell surface, in the presence of a known amount of detectably labeled ezetimibe, with a sample to be tested for the presence of the antagonist;
- and
- (b) measuring the amount of detectably labeled ezetimibe specifically bound, directly or indirectly, to the polypeptide;
- wherein an NPC1L1 antagonist in the sample is identified by measuring substantially reduced direct or indirect binding of the detectably labeled ezetimibe to the polypeptide, compared to what would be measured in the absence of such an antagonist.
A method for identifying an antagonist of NPC1L1 comprising:
- (a) placing, in an aqueous suspension, a plurality of support particles, impregnated with a fluorescer, to which a host cell expressing a polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 2, 4 and 12 or a functional fragment thereof on a cell surface are attached;
- (b) adding, to the suspension, radiolabeled ezetimibe and a sample to be tested for the presence of the antagonist, wherein the radiolabel emits radiation energy capable of activating the fluorescer upon direct or indirect binding of the ezetimibe to the polypeptide to produce light energy, whereas radiolabeled ezetimibe that does not directly or indirectly bind to the polypeptide is, generally, too far removed from the support particles to enable the radioactive energy to activate the fluorescer;
- and
- (c) measuring the light energy emitted by the fluorescer in the suspension;
- wherein an NPC1L1 antagonist in the sample is identified by measuring substantially reduced light energy emission, compared to what would be measured in the absence of such an antagonist.
A method for identifying an antagonist of NPC1L1 comprising:
- (a) contacting a host cell expressing a polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 2, 4 and 12 or a functional fragment thereof on a cell surface with a detectably labeled sterol or 5α-stanol and with a sample to be tested for the presence of the antagonist;
- and
- (b) measuring the amount of detectably labeled sterol or 5α-stanol in the cell;
- wherein an NPC1L1 antagonist in the sample is identified by measuring substantially reduced detectably labeled sterol or 5α-stanol within the host cell, compared to what would be measured in the absence of such an antagonist.
A mutant mouse comprising a homozygous mutation of endogenous, chromosomal NPC1L1 wherein the mouse does not produce any functional NPC1L1 protein.
A kit comprising:
- (a) ezetimibe in a pharmaceutical dosage form;
- and
- (b) information indicating that NPC1L1 is a target of ezetimibe.
A method for decreasing the level of intestinal sterol or 5α-stanol absorption in a subject comprising reducing the level of expression of NPC1L1 in the subject.
A method for identifying an antagonist of NPC1L1 comprising:
- (a) contacting a host cell expressing a polypeptide comprising an amino acid sequence selected from SEQ ID NOs:
- 2, 4 and 12 or a functional fragment thereof on a cell surface, in the presence of a known amount of a detectably labeled 2-azetidinone, with a sample to be tested for the presence of the antagonist;
- and
- (b) measuring the amount of detectably labeled 2-azetidinone specifically bound, directly or indirectly, to the polypeptide;
- wherein an NPC1L1 antagonist in the sample is identified by measuring substantially reduced direct or indirect binding of the detectably labeled 2-azetidinone to the polypeptide, compared to what would be measured in the absence of such an antagonist.
A kit comprising:
- (a) a 2-azetidinone in a pharmaceutical dosage form;
- and
- (b) information indicating that NPC1L1 is a target of the 2-azetidinone.
Dependencies are inferred from the claim text.