Polymorphisims in the human cyp2d6 gene promoter region and their use in diagnostic and therapeutic applications

Abstract

Provided are polynucleotides of molecular variant promoters of the CYP2D6 gene which, for example, are associated with abnormal drug response or individual predisposition to several common diseases and disorders caused by drug under- or over-metabolization, and vectors comprising such polynucleotides. Furthermore, methods of diagnosing the status of disorders related to intermediate metabolization of drugs are described. In addition, kits comprising oligonucleotides hybridizing to the CYP2D6 promoter and/or being capable of being extended into this region useful for diagnosing subjects that are ultrarapid or intermediate metabolizer of drugs are provided.

1 . A polynucleotide comprising a polynucleotide selected from the group consisting of:
 (a) a molecular variant polynucleotide having the nucleic acid sequence of SEQ ID NO: 1, wherein at nucleotide position corresponding to nucleotide position −1584 of the CYP2D6 promotor as shown in  FIG. 1 , is a G;   (b) a molecular variant polynucleotide capable of hybridizing to the CYP2D6 promotor as shown in  FIG. 1 , wherein said polynucleotide is having at a position corresponding to position −1584 of the CYP2D6 promoter as shown in  FIG. 1  at least one nucleotide deletion, addition and/or substitution; and   (c) a molecular variant polynucleotide capable of hybridizing to the CYP2D6 promoter as shown in  FIG. 1 , wherein said polynucleotide is having at a position corresponding to position −1584 of the CYP2D6 promotor as shown in  FIG. 1 , a G.   
     
     
         2 . The polynucleotide of  claim 1 , wherein the nucleotide deletion, addition and/or substitution result(s) in altered expression of the variant CYP2D6 gene compared to the corresponding wild type gene. 
     
     
         3 . A vector comprising the polynucleotide of  claim 1  or  2 . 
     
     
         4 . A host cell genetically engineered with the polynucleotide of  claim 1  or  2  or the vector of  claim 3 . 
     
     
         5 . A method of diagnosing a disorder related to a reduced or enhanced capacity for clearance of CYP2D6 substrates or susceptibility to such a disorder comprising determining the presence of a mutation in the CYP2D6 promoter in a sample from a subject. 
     
     
         6 . The method of  claim 5 , wherein said substrate is selected from antiarrhythmics, beta adrenergic receptor antagonists, tricyclic antidepressants, selective serotonin reuptake inhibitors (SSRI), neuroleptics, opiates, anticancer agents or amphetamines. 
     
     
         7 . The method of  claim 5  or  6  comprising PCR, ligase chain reaction, restriction digestion, direct sequencing, nucleic acid amplification techniques or hybridization techniques. 
     
     
         8 . The method of any one of  claims 5  to  7 , further comprising administering to a subject a medicament to abolish or alleviate said disorder. 
     
     
         9 . Use of an oligo- or polynucleotide for the detection of a polynucleotide of  claim 1  or  2  and/or for genotyping of individual CYP2D6 promoter alleles. 
     
     
         10 . The use of  claim 9 , wherein said polynucleotide is a polynucleotide of  claim 1  or  2 . 
     
     
         11 . The use of  claim 9 , wherein said oligonucleotide is about 15 to 50 nucleotides in length and comprises the nucleotide sequence of any one of SEQ ID NOs: 2 to 10 or a complementary sequence. 
     
     
         12 . A primer or probe consisting of an oligonucleotide as defined in  claim 11 . 
     
     
         13 . A kit useful for carrying out a method of any one of  claims 5  to  9  comprising oligonucleotides or polynucleotides capable of detecting the presence of a polynucleotide of  claim 1  or  2 , and optionally suitable means for detection. 
     
     
         14 . Use of an effective dose of a drug or prodrug for the preparation of a pharmaceutical composition for the treatment or prevention of a disorder of a subject comprising a polynucleotide of  claim 1  or  2  in its genome.