METHOD OF TESTING DRUG FOR TREATING OR PREVENTING DISEASES SUCH AS HYPERLIPEMIA

摘要

Testing drugs that have activity on treating or preventing at least 1 disease selected from hyperlipemia, arteriosclerosis and hyperglycemia comprising culturing cells originating from mammal in the presence or absence of a test substance, and detecting expression dose of the mRNA with any of the specified nucleotide sequences, is new. Testing drugs that have activity on treating or preventing at least 1 disease selected from hyperlipemia, arteriosclerosis and hyperglycemia comprising: (a) culturing cells originating from mammal in the presence or absence of a test substance; (b) detecting expression dose of the mRNA with any of the nucleotide sequences (a)-(e) (where t and u are exchangeable): (i) nucleotides 47-1411 of a 1604 base pair sequence (I), given in the specification; (ii) nucleotides 78-1457 of a 1716 base pair sequence (III), given in the specification; (iii) the DNA inserted with a phagemid sustaining in the transformant E. coli pBK/m55-1 SANK 72199 (FERM BP-6940); (iv) the DNA inserted with a phagemid sustaining in the transformant E. coli pTrip/h55-1 SANK 72299 (FERM BP-6941); or (v) a nucleotide sequence hybridizable with a polynucleotide containing the antisense sequence of (a)-(d) under stringent conditions and encoding a polypeptide with the activity of increasing neutral lipid concentration in serum; and (c) comparing the resultant expression doses for selecting a test substance. Independent claims are also included for: (1) a similar method for testing substances for treating or preventing hyperlipemia, arteriosclerosis or/and hyperglycemia by using an antibody binding specifically to the polypeptide containing an amino acid sequence or a part of it encoded by any of the nucleotide sequences (i)-(v) in order to quantify its amount produced after culturing with the test substance, during which the amount of the polypeptide thus detected is compared with that obtained in a control run; (2) a kit for testing the remedies or preventives for hyperlipemia, arteriosclerosis or/and hyperglycemia containing any of the antibodies specific for the polypeptides or their partial peptides; (3) compositions for gene therapy of the diseases comprising DNA or RNA with an antisense sequence of a nucleotide sequence composed of 15-30 consecutive nucleotides of residues 78-145 of (III); (4) a polypeptide containing 2 or more consecutive residues of a 207 (XII) or 406 (XIV) amino acid sequence (XII), given in the specification; (5) a polypeptide with a 491 (XVIII), 493 (XX), 470 (XXII), 346 (XXIV) or 215 (XXVI) residue amino acid sequence, given in the specification; (6) DNAs encoding the polypeptides of (4) or (5); (7) a recombinant vector, particularly an adenovirus vector, containing the DNA of (6); (8) a host cell transformed with the recombinant vector of (7); (9) a process for producing the biologically-active polypeptide by culturing the host cell of (8) before recovering the product from the cultured material; (10) testing remedies or preventives of the specified diseases comprising: (a) administering 1 or more of the polypeptides and test substance to a non-human mammal, and (b) measuring neutral lipid concentration in blood of such animal; (11) testing remedies or preventives of the specified diseases comprising: (a) infecting a non-human mammal with the recombinant adenovirus vector-sustaining adenovirus; (b) administering a test substance to the animal; and (c) measuring the neutral lipid concentration in blood of the animal; (12) testing remedies or preventives of the specified diseases comprising mixing lipoprotein lipase (LPL) and/or hepatic triacylglycerol lipase (HTGL) as substrate in a reaction system with the test substance in the presence of a polypeptide containing an amino acid sequence from positions 17-460 in the sequence of (IV), a polypeptide lacking the residues 1-253 in its carboxy terminal, or any of the polypeptides e.g. based on amino acid sequences (XII)-(XXVI), and investigating inhibitory effect of the test substance on the inhibition of LPL and/or HTGL activity by the polypeptide; (13) testing substances modulating the LPL activity comprising: (a) preparing mixtures of materials containing LPL, its substrate and the polypeptide based on the amino acid sequence of (IV), or (XII)-(XXVI) with the test substance; (b) combining them; and (c) measuring the amounts of fatty acid liberated from the substrate for comparison; (14) DNA containing, or having, a nucleotide sequence from positions 1-510, or 1-526 of a 526 nucleotide sequence (XXVII), given in the specification; (15) an expression vector containing the DNA derived from the nucleotide sequence of (XXVII), or expressing a foreign gene in mammalian cells obtained by ligating the DNA in the same transcription direction as that of the foreign sequence at upstream side of the foreign gene; (16) recombinant plasmid pGL8-3 (FERM BP-7627) and expression vector pGL11-4; (17) a host cell transformed with the plasmid or vector of (16); (18) screening remedies or preventives of the specified diseases comprising culturing a test substance with the host cells and measuring the amount of expressed foreign protein; (19) screening remedies or preventives of the specified diseases by detecting promoter activity, or protein production level of angiopoetin-associated protein 3 or 4 gene in the nuclear extract of mouse cells with measurement of the radiolabeled DNA after contact with a test substance and separation using electrophoresis; (20) a DNA containing a nucleotide sequence with not less than 10 consecutive nucleotides from positions 1-526 in (XXVII), a nucleotide sequence contains its antisense sequence, or their inductants; and (21) DNA containing a nucleotide sequence with not less than 10 consecutive nucleotides from positions 1-260 of a 260 base pair sequence (XXVIII), given in the specification, a nucleotide sequence contains its antisense sequence, a nucleotide can promote or inhibit the promoter activity of the DNA with a sequence of (XXVIII) and has promoter activity, or their inductants. ACTIVITY : Antilipemic; Antiarteriosclerotic; Antidiabetic. No biological data is given. MECHANISM OF ACTION : Gene therapy.