Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
  • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
  • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q2600/00—Oligonucleotides characterized by their use
  • C12Q2600/156—Polymorphic or mutational markers

Definitions

• test sample of the nucleic acid carrying the said polymorphism is conveniently a sample of blood, bronchoalveolar lavage fluid, sputum, urine or other body fluid or tissue obtained from an individual.
• test sample may equally be a nucleic acid sequence corresponding to the sequence in the test sample, that is to say that all or a part of the region in the sample nucleic acid may firstly be amplified using any convenient technique, e.g. polymerase chain reaction (PCR) or ligase chain reaction (LCR), before analysis of allelic variation.
• PCR polymerase chain reaction
• LCR ligase chain reaction
• the invention relates to allele-specific oligonucleotide probes for detecting a polymorphism in the UGTl gene capable of hybridizing to diagnostic nucleic acids comprising within their sequence the polymorphisms as defined above.
• the diagnostic kits may comprise appropriate packaging and instructions for use in the methods of the invention. Such kits may further comprise one or more appropriate buffers and one or more polymerases such as thermostable polymerases, for example Taq polymerase. Such kits may also comprise companion/constant primers and/or control primers or probes. A companion/constant primer is one that is part of the pair of primers used to perform PCR. Such primer usually complements the template strand precisely. Furthermore the invention relates to a pharmaceutical pack comprising a pharmaceutically active compound like Tolcapone and instructions for administration of the drug to human beings diagnostically tested for a single nucleotide polymorphism according to a method of the present invention.
• a pharmaceutically active compound like Tolcapone
• Figure 1 shows the primary metabolic routes of tolcapone in the liver.
• Tolcapone is oxidized by cytochrome P450 3A4 (CYP3A4), the nitro group is reduced and acetylated by N-acetyltransferase (NAT).
• the phenolic hydroxy group can be sulfated by sulfo transferase (ST) or methylated by catechol-O- methyl transferase (COMT).
• ST sulfo transferase
• COMP catechol-O- methyl transferase
• Glucuronidation of the hydroxy group a major reaction of detoxification in the liver, is catalyzed by UDP- glucuronosyltransferase (UGT). Subsequent oxidation or conjugation with glucuronate, sulphate and acetate further modifies primary metabolites.
• UDP- glucuronosyltransferase UDP- glucuronos
• FIG. 2 represents the UGTlA gene structure.
• the UGTlA gene spans more than
• Amplification reactions were prepared using an aliquoting robot (Packard Multiprobe II, Meriden, CT) in 96- well amplification plates identified by barcode labels generated by the experiment management database. Parameters for procedures performed by the robot were set to minimize the possibility of cross-contamination. For each plate of 81 samples, 5 samples were run in duplicate and the duplicate results were analysed to determine that they matched.
• Packard Multiprobe II Meriden, CT
• UGTl-ex5-l fragment UGTlex5-l-F CAGTTAGCCATGCTTGTGCC (SEQ ID N0:51)
• Primer UGTlex5-l-F corresponds to positions 63 to 82 in exon 5 of UGTl as defined by the positions in SEQ ID NO:l.
• Primer UGTlex5-l-R hybridizes to positions 684 to 703 as defined by the positions in SEQ ID NO:l.
• Primer UGTlex5-2-F corresponds to positions 461 to 480 in exon 5 of UGTl as defined by the positions in SEQ ID NO:l.
• Primer UGTlex5-2-R hybridizes to positions 1082 to 1101 as defined by the positions in SEQ ID NO:l.
• the genetic markers were selected based on the known pharmacology of tolcapone and knowledge from the literature of genetic polymorphisms that could affect the activity of corresponding and relevant gene products.
• the main metabolic pathway for tolcapone elimination is glucuronidation by UGTl enzymes.
• UGTlA10exonl_959. The number refers to the position of the SNP relative to the DNA sequence with Genbank accession number U39550 from the public database.

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• Chemical & Material Sciences (AREA)
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• General Engineering & Computer Science (AREA)
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• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

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• 2001
  • 2001-07-02 WO PCT/EP2001/007524 patent/WO2002006523A2/en not_active Ceased
  • 2001-07-02 US US10/333,108 patent/US20040076968A1/en not_active Abandoned
  • 2001-07-02 EP EP01960438A patent/EP1325152A2/en not_active Withdrawn
  • 2001-07-02 AU AU2001281930A patent/AU2001281930A1/en not_active Abandoned
  • 2001-07-02 JP JP2002512413A patent/JP3947103B2/ja not_active Expired - Fee Related

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