KR101760246B1 - Novel SLCO2B1 Single Nucleotide Polymorphism Markers and Uses Thereof - Google Patents

Abstract

The present invention relates to a novel single nucleotide polymorphism (SNP) present in the gene of SLCO2B1 (Solute carrier organic anion transporter family member 2B1) and its use as a marker for predicting drug response, and more particularly to a SLCO2B1 single nucleotide polymorphism A drug reaction prediction or diagnosis kit, a method of providing information for drug selection determination, a method for detecting a mutation genotype of a gene associated with a drug reaction, and a method for providing information on a single nucleotide polymorphism of the SLCO2B1 gene will be. The novel SLCO2B1 single nucleotide polymorphism markers presented in the present invention can be useful as biomarkers related to drug transporter dysfunction. That is, the novel SLCO2B1 single nucleotide polymorphism of the present invention can be used as a biomarker to predict or diagnose abnormalities due to drug transporter activity deficiency and drug transporter deficiency. In particular, the SLCO2B1 gene single nucleotide polymorphism of the present invention is a novel mutation gene discovered in Koreans and can contribute to the development of genetic diagnostic methods for Koreans by providing core base information in the development of a customized drug treatment method for Koreans.

Description

Novel SLCO2B1 polymorphism markers and their uses {Novel SLCO2B1 Single Nucleotide Polymorphism Markers and Uses Thereof}

The present invention relates to a novel single nucleotide polymorphism (SNP) present in the gene of SLCO2B1 (Solute carrier organic anion transporter family member 2B1) and its use as a marker for predicting drug response, and more particularly to a SLCO2B1 single base A method for predicting or diagnosing drug response using polymorphism, a kit for predicting or diagnosing drug response, a method for providing information for drug selection determination, a method for detecting a mutation genotype of a gene related to a drug reaction, and a method for providing information on single nucleotide polymorphisms of the SLCO2B1 gene .

It is known that 99.9% of human genes are identical in all individuals regardless of race, only 0.1% are individual differences, and this 0.1% gene difference determines a large part of human characteristics or appearance. Skin color, baldness, hair color, appearance of key, diabetes, dementia, cancer and other diseases are various phenomena because of their different genetic information.

The response to drugs prescribed by the hospital can vary from person to person and is known to be influenced by genetic factors. Drugs administered for therapeutic purposes may exhibit the desired therapeutic effect in the desired direction, but may exhibit unexpected adverse drug effects or lack therapeutic effects. Drug genomics is a discipline that studies how genes determine the diversity of drug responses and how different genes and drug responses change depending on individuals.

Drug-genomics research is a core technology that realizes the era of 'customized drug therapy' that can predict what kind of drugs, what dose, who, when, and when, even if only the patient's genetic information is known in the near future . Drugs prescribed by a doctor are not always effective for all patients, and the effect of the drug varies significantly among individual patients. Among patients who have been properly prescribed and prescribed by the physician and who have taken the medication properly, there are often life-threatening adverse drug reactions (such as drug and drug side effects) often occurring, (Source: http://www.kosen21.org/nwebzine/webzine_view.jsp?webzine_seq=76&board_seq=899&data_seq=1186).

Single nucleotide polymorphism (SNP) is the most common form of human gene mutation. One of the nucleotide sequences consisting of A, T, C, and G in the genome has a different base sequence It means change. Two-thirds of these SNPs are known to be mutations in the nucleotide sequence between C and T, and it is known that SNP mutations usually occur once per 1000 nucleotides in the genomic sequence. In addition, SNPs account for about 90% of mutations in human genomes, and people with similar traits or families have the same or similar SNP patterns, so clinical susceptibility to disease in individuals And can also be used as an indicator for predicting the effects and side effects of drugs. Furthermore, SNPs can be used in personalized medicine that provides diagnostic and therapeutic strategies tailored to the genetic character of the individual, and can be used to construct medical statistics for the treatment of various diseases if SNP patterns and disease records can be well integrated There will be.

Recently, the use of gene polymorphism as an index for predicting drug response in individuals has been developed, and in particular, personalized treatment is being actively studied.

In particular, since Korean drug genome information differs from other species, it is necessary to systematically construct a mutation map in order to develop customized therapeutic technology for Koreans. The mutation of the drug genome in Koreans is also recognized as a state-of-the- It is important to obtain information on mutation in order to develop genetic diagnostic methods for Koreans.

SLCO2B1 is a member of the Solute carrier organic anion transporter family member. It is also known as OATP-B, OATP2B1, SLC21A9 and is expressed in various organs such as liver, lung, kidney and small intestine. (Ikumi Tamai et al., 2006). In particular, SLCO2B1, which is mainly expressed in the sinusoidal membrane of hepatocytes, is involved in absorption into hepatocytes. In addition, atorvastatin, which inhibits hydroxymethylglutaryl-CoA (HMG-CoA) reductase and lowers LDL-cholesterol levels, is also a substrate drug of OATP and has a high affinity for OATP2B1 in vitro . It has also been reported that OTRP inhibits the transport of estrone-3-sulfate, an endogenous substrate of OATP (Br J Pharmacol 2009).

Accordingly, the present inventors have uncovered a novel mutant gene of a Korean specific marker gene related to the dysfunction of the SLCO2B1 drug transport protein through the next generation sequencing analysis of the Disease Control Headquarters and the Genome Service Agency, and conducted an experiment using this novel mutation type expression vector As a result, the present inventors have completed the present invention by confirming that the function of the drug transport in comparison with the wild type is decreased or increased in the case of cells into which the mutant SLCO2B1 gene has been introduced.

Korean Patent No. 10-0677724 Korean Patent Publication No. 10-2007-0111475 Korean Patent Publication No. 10-2010-0077396

Accordingly, an object of the present invention is to provide a composition which can effectively predict (diagnose) a drug response using a single base polymorphism of SLCO2B1 gene as a specific marker of Korean.

It is still another object of the present invention to provide a kits for Korean specific drug response prediction (diagnosis) including the above composition.

It is another object of the present invention to provide a method for providing information for Korean specific drug selection decisions.

It is another object of the present invention to provide a method for detecting a Korean specific mutation type of SLCO2B1 gene related to drug reaction.

Yet another object of the present invention is to provide a method for providing information on single nucleotide polymorphisms of the Korean specific SLCO2B1 gene.

In order to achieve the above-mentioned object of the present invention,

The present invention relates to a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene (reference sequence NM_007256.4) and a polynucleotide comprising the polynucleotide of SEQ ID NO: 1 as a single base of a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 An agent capable of detecting polymorphism (SNP); Or an amino acid selected from the group consisting of 271, 381, 400, and 642 th amino acids in the polypeptide consisting of SEQ ID NO: 2.

In one embodiment of the present invention, the 1207th base is G or T; The 1536th base is G or T; The 1594th base is G or T; Or the 2319 < th > base may be T or C. [

In one embodiment of the present invention the drug is selected from the group consisting of glibenclamide, estrone-3-sulfate, benzylpenicillin, fexofenadine, Bosentan, statin, pravastatin pravastatin, fluvastatin, atorvastatin, and rosuvastatin.

In one embodiment of the present invention, the agent capable of detecting the single nucleotide polymorphism (SNP) comprises a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: 1 Of a SNP, or a primer capable of amplifying a complementary polynucleotide of the polynucleotide, which is composed of 10 to 100 contiguous bases.

In one embodiment of the present invention, the agent capable of detecting the single nucleotide polymorphism (SNP) comprises a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: 1 And a probe capable of hybridizing with a polynucleotide consisting of 10 to 100 contiguous bases including the SNP of the probe.

In one embodiment of the present invention, the agent capable of detecting the amino acid can form a bond with an amino acid selected from the group consisting of 271st, 381st, 400th and 642nd amino acids in the polypeptide consisting of SEQ ID NO: 2 Lt; / RTI >

The present invention also provides a kit for predicting or diagnosing a drug response comprising the composition.

(A) separating a sample from a specimen; And (b) identifying a base selected from the group consisting of 1207th, 1536th, 1594th and 2319th bases in the polynucleotide of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene in the separated sample, or Identifying amino acids selected from the group consisting of 271, 381, 400 and 642 th SLCO2B1 proteins in the polypeptide consisting of SEQ ID NO: 2.

In one embodiment of the present invention, when the 1207th base of SEQ ID NO: 1 is replaced with a thymine (T) in guanine (G) as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the polypeptide of SEQ ID NO: 2 The 271st amino acid may be a mutant in which arginine is substituted with leucine.

In one embodiment of the present invention, when the 1536th base of SEQ ID NO: 1 is replaced with a thymine (T) in guanine (G) as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the polypeptide of SEQ ID NO: 2 The 381st amino acid may have a variant in which the valine is substituted with leucine.

In one embodiment of the present invention, when the 1594th base of SEQ ID NO: 1 is replaced with thymine (T) in guanine (G) as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the polypeptide of SEQ ID NO: 2 And the 400th amino acid may be substituted with leucine at arginine.

In one embodiment of the present invention, when the 2319th base in SEQ ID NO: 1 is replaced with a cytosine (C) in a thymine (T) as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, The 642nd amino acid may have a variant in which the histidine is substituted in the tyrosine.

In one embodiment of the present invention, when the 381th amino acid of the polypeptide consisting of SEQ ID NO: 2 has a variant in which the valine is substituted with leucine, it can be determined that the drug absorption is inhibited.

In one embodiment of the present invention, the 271th amino acid of the polypeptide consisting of SEQ ID NO: 2 is substituted with leucine in arginine, the mutant in which the 400th amino acid is substituted with leucine in arginine or the 642th amino acid is substituted with histidine , It can be judged that the drug absorption is promoted.

In one embodiment of the present invention the drug is selected from the group consisting of glibenclamide, estrone-3-sulfate, benzylpenicillin, fexofenadine, Bosentan, statin, pravastatin pravastatin, fluvastatin, atorvastatin, and rosuvastatin.

In one embodiment of the present invention, when the 381st amino acid of the polypeptide consisting of SEQ ID NO: 2 has a variant in which the valine is substituted with leucine, drug absorption of the estrone-3-sulfate drug is inhibited, Estrone-3-sulfate drugs may not be prescribed in the case of a sample having such a substituted variant.

In addition, the present invention relates to a nucleic acid sample isolated from a sample, wherein a base selected from the group consisting of 1207th, 1536th, 1594th and 2319th bases in the polynucleotide of SEQ ID NO: 1 as a single base polymorphism (SNP) region of the SLCO2B1 gene Checking; Or identifying the amino acid selected from the group consisting of 271, 381, 400 and 642 in the polypeptide consisting of SEQ ID NO: 2 in the protein sample isolated from the sample. .

In one embodiment of the present invention, the mutated genotype of SLCO2B1 is a polynucleotide of SEQ ID NO: 1 wherein the 1207th base is substituted with guanine (G) to thymine (T); The 1536th base is replaced by thymine (T) in guanine (G); The 1594th base is replaced with thymine (T) in guanine (G); And at least one mutation selected from the group wherein the 2319 base is substituted with cytosine (C) from thymine (T).

The present invention also provides a method for providing information on the single nucleotide polymorphism of the SLCO2B1 gene, comprising the step of identifying a base selected from the group consisting of the 1207th, 1536th, 1594th and 2319th bases of the SLCO2B1 gene of SEQ ID NO: 1 to provide.

In one embodiment of the present invention, the method further comprises determining whether the 1207th base of the SLCO2B1 gene in the polynucleotide of SEQ ID NO: 1 is substituted with guanine (G) to thymine (T); Whether or not the 1536th base of the SLCO2B1 gene of SEQ ID NO: 1 is replaced with thymine (T) in guanine (G); Whether the 1594th base of the SLCO2B1 gene of SEQ ID NO: 1 was substituted with thymine (T) in guanine (G); Or confirming whether the 2319th base of the SLCO2B1 gene of SEQ ID NO: 1 has been substituted with cytosine (C) in thymine (T).

The novel SLCO2B1 single nucleotide polymorphism markers presented in the present invention can be useful as biomarkers related to drug transporter dysfunction. That is, the novel SLCO2B1 single nucleotide polymorphism of the present invention can be used as a biomarker to predict or diagnose abnormalities due to drug transporter activity deficiency and drug transporter deficiency. In particular, the SLCO2B1 gene single nucleotide polymorphism of the present invention is a novel mutation gene discovered in Koreans and can contribute to the development of genetic diagnostic methods for Koreans by providing core base information in the development of a customized drug treatment method for Koreans.

1 shows SLCO2B1 271L type, SLCO2B1 381L type, SLCO2B1 400L type, and SLCO2B1 642H type variant from the left.
Figure 2 shows the drug transport activity of the wild-type and mutant SLCO2B1 protein in terms of the number of decays of radioactive elements per minute (dpm).
Figure 3 shows the results of the enzymatic analysis of wild-type and mutant SLCO2B1 proteins.
Figure 4 is a western blot result of a protein expressed by wild type and mutant SLCO2B1 cDNA (lane 1: negative control (pcDNA3.1 +), lane 2: wild type SLCO2B1, lane 3: mutated SLCO2B1 Arg271Leu, lane 4: mutation Type SLCO2B1 Val381Leu, lane 5: mutant SLCO2B1 Arg400Leu, lane 6: mutant SLCO2B1 Tyr642His).
FIG. 5 shows the membrane potential protein pattern of SLCO2B1, and the single base polymorphism is shown in red.
Fig. 6 shows a vector map of the expression vector pcDNA 3.1 (+) used in this example.

The present invention relates to a single nucleotide polymorphism at the 1207th, 1536th, 1594th or 2319th base position of the SLCO2B1 gene consisting of the polynucleotide of SEQ ID NO: 1; And a use thereof for predicting or diagnosing a drug reaction using a biomarker as a biomarker.

More specifically, the single nucleotide polymorphism in the SLCO2B1 gene consisting of the polynucleotide of SEQ ID NO: 1 indicates that the 1207th base is G or T; 1536 < th > base is G or T; 1594 base is G or T; Or the 2319 < th > base may be represented by T or C. [

SLCO2B1 is a member of the Solute carrier organic anion transporter family member. It is a protein involved in absorption of many drugs as well as various endogenous substances, and is expressed in various organs such as liver, lung, kidney, small intestine.

The present inventors conducted a study on drug genetic polymorphisms targeting Korean people and searched for genetic mutations associated with Korean drug reactions distinct from other species. As a result, it was found that the next generation sequencing (R271L, V381L, R400L, Y642H) of the mutant SLCO2B1 gene identified in the nonspecific gene (NGS). The single nucleotide polymorphism (SNP) sites of the four new SLCO2B1 mutation genes (R271L, V381L, R400L and Y642H) are 1207G> T, 1536G> T, 1594G> T and 2319T> C, respectively.

As a result, it was confirmed that SLCO2B1 mutant genes R271L, R400L and Y642H of the present invention have a carrier (transport) function of the SLCO2B1 protein, and when carrying the SLCO2B1 mutant gene V381L, the SLCO2B1 protein carrier It was confirmed that it was inhibited.

The variation described in this specification will be described as follows.

For example, the " R271L " mutation means that the 271st amino acid in the amino acid sequence of the SLCO2B1 protein consisting of SEQ ID NO: 2 is substituted with arginine (R) to leucine (L), and the " 1207G> T " mutation means SLCO2B1 And the 1207th base in the nucleotide sequence of the gene is replaced with guanine (G) to thymine (T).

Thus, the SLCO2B1 mutant gene R271L, V381L, R400L or Y642H of the present invention is a variant in which the 271st amino acid in the amino acid sequence of the SLCO2B1 protein consisting of SEQ ID NO: 2 is substituted with arginine (R) to leucine (L); A variant in which the 381st amino acid is substituted with leucine (L) in valine (V); A variant in which the 400th amino acid is replaced by leucine (L) in arginine (R); And the 642th amino acid is substituted with histidine (H) in tyrosine (Y).

Also, each of the SLCO2B1 mutation genes R271L, V381L, R400L or Y642H can be represented as a single nucleotide polymorphism (SNP) by 1207G> T, 1536G> T, 1594G> T or 2319T> Wherein 1207G > T is a polynucleotide consisting of the polynucleotide SLCO2B1 gene consisting of SEQ ID NO: 1 in which the 1207th base is substituted with thymine (T) in guanine (G); Mutation in which the 1536th base is replaced with thymine (T) in guanine (G); Mutation in which the 1594th base is replaced with thymine (T) in guanine (G); Refers to a variant in which the 2319 base is substituted with cytosine (C) in thymine (T).

Accordingly, the present invention relates to a polynucleotide comprising a polynucleotide of SEQ ID NO: 1 as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, a single nucleotide polymorphism (SNP) of a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 A detectable agent; Or an amino acid selected from the group consisting of 271, 381, 400, and 642 th amino acids in the polypeptide consisting of SEQ ID NO: 2.

In one embodiment of the present invention, the 1207th base may be G or T; The 1536th base may be G or T; The 1594th base may be G or T; The 2319 < th > base may be T or C. [

In the present invention, the term " for predicting or diagnosing a drug response " means an application for predicting or diagnosing the degree of absorption of a drug, which may appear when a drug is applied to an animal including a human being.

The SLCO2B1 protein of the present invention corresponds to a protein associated with various types of drug absorption (transport), and the drug absorption efficiency may vary depending on the SLCO2B1 gene or protein function abnormality.

Examples of the SLCO2B1 protein-mediated drug include glibenclamide, estrone-3-sulfate, benzylpenicillin, fexofenadine, Bosentan, statin, pravastatin, ), Fluvastatin, atorvastatin and rosuvastatin. However, the type of the drug is not particularly limited as long as it is a drug involved in SLCO2B1 protein.

In detail, glibenclamide corresponds to a therapeutic agent for diabetes, and SLCO2B1 protein is known to be involved in absorption of the above-mentioned glibenclamide drug; Estrone-3-sulfate is a natural steroid preparation, which is applied to various diseases of postmenopausal women. SLCO2B1 protein is known to be involved in the absorption of estrone drugs; Benzylpenicillin is an antibiotic and SLCO2B1 protein is known to be involved in the absorption of the antibiotic; Fexofenadine is an antihistamine and the SLCO2B1 protein is known to be involved in the absorption of the fexofenadine drug; Bosentan is an antihypertensive agent and SLCO2B1 protein is known to be involved in the absorption of the above-mentioned botanic drugs; Statin is known to be a therapeutic agent for hyperlipemia. SLCO2B1 protein is known to be involved in the uptake of statin drugs; In addition, pravastatin, fluvastatin, atorvastatin, and rosuvastatin are statin drugs that inhibit HMG-CoA reductase, an enzyme involved in the synthesis of cholesterol, and SLCO2B1 protein It is also known to be involved in absorption of drugs.

Therefore, when the SLCO2B1 function abnormality is present, the absorption efficiencies of the above exemplified drugs can be changed. As a result, the novel SLCO2B1 mutant genes (R271L, V381L, R400L, Y642H) And thus can greatly contribute to prevention of misuse of drugs in selecting future drugs.

In the following examples of the present invention, the drug-transferring ability of each of four mutant SLCO2B1 was tested using estrone-3-sulfate as a representative drug transported by SLCO2B1 protein. As a result, R271L, R400L and Y642H of SLCO2B1 gene In the case of the mutant type, it was confirmed that the drug delivery was more effective than the normal drug delivery, and that the V381L mutant did not normally carry the drug.

These results can be used as basic data for the determination of estrone drug selection.

For example, in the case of patients with the R271L, R400L, Y642H, and V381L variants of the SLCO2B1 gene after discrimination of individuals with single nucleotide polymorphisms that require prescription of estrone drugs for the treatment of postmenopausal women, As the absorption efficiency changes, the same therapeutic effect can be expected with the coordinated dose to the standard dose of the prescribed drug. If the desired pharmacological effect can not be achieved, an alternative drug can be prescribed instead of the estrone drug. Can be used effectively.

The agent capable of detecting the single nucleotide polymorphism (SNP) contained in the composition of the present invention is a polynucleotide comprising a nucleotide sequence selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 of the polynucleotide of SEQ ID NO: 1 A primer capable of amplifying a polynucleotide consisting of 10 to 100 contiguous bases comprising a SNP or a complementary polynucleotide thereof; Or a probe capable of hybridizing with a polynucleotide consisting of 10 to 100 contiguous bases including the SNP.

The polynucleotide or its complementary polynucleotide according to the present invention is a polymorphic sequence. A polymorphic sequence refers to a sequence comprising a polymorphic site representing a single base polymorphism in the nucleotide sequence. A polymorphic site is a site in a polymorphic sequence where a single base polymorphism occurs.

The polynucleotide or its complementary polynucleotide according to the present invention has a polymorphic sequence. In the present invention, the polynucleotide may be DNA. The polynucleotide may preferably be composed of 10 or more, more preferably 10 to 100, even more preferably 10 to 60 contiguous bases.

In the present invention, a primer or a probe that specifically hybridizes with the polynucleotide or a complementary polynucleotide thereof is specific to a single nucleotide polymorphism (SNP).

Single nucleotide polymorphism (SNP) specific refers to hybridizing specifically to each SNP, that is, hybridizing so that the nucleotide at a polymorphic site present in the polymorphic sequence can be specifically discriminated. Here, hybridization is usually carried out under stringent conditions, for example, a salt concentration of 1 M or less and a temperature of 25 ° C or higher.

In the present invention, a probe means a hybridization probe, and means an oligonucleotide capable of binding sequence-specifically to a complementary strand of a nucleic acid. Preferably, the probe of the present invention aligns with the polymorphic site of the polymorphic sequence. The probe of the present invention can be used in a kit or a prediction method such as a microarray for predicting or diagnosing a drug reaction by detecting a single base polymorphic site.

Further, in the present invention, the primer can form base pairs with a complementary template with a base sequence having a short free 3 'hydroxyl group, and can be used as a starting point for template strand copy Quot; short sequence " The appropriate length of the primer may vary depending on the purpose of use, but is generally comprised of 15 to 30 bases. The primer sequence need not be completely complementary to the template, but should be sufficiently complementary to hybridize with the template. The primers can be hybridized to a DNA sequence containing a polymorphic site to amplify a DNA fragment containing the polymorphic site. A kit or a prediction method such as a microarray for predicting or diagnosing a drug reaction by detecting a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene of the present invention.

The primers or probes of the present invention can be chemically synthesized using the phosphoramidite solid support method, or other well-known methods. Such nucleic acid sequences may also be modified using many means known in the art. Non-limiting examples of such modifications include, but are not limited to, methylation, capping, substitution of one or more natural nucleotides with an analogue, and modification between nucleotides, such as uncharged linkers (e.g., methylphosphonate, phosphotriester, Amidates, carbamates, etc.) or charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.).

In addition, the agent capable of detecting the amino acid contained in the composition of the present invention according to the present invention comprises an amino acid selected from the group consisting of 271st, 381st, 400th and 642nd amino acids in the polypeptide consisting of SEQ ID NO: Lt; RTI ID = 0.0 > antigen-antibody < / RTI >

In another aspect, the present invention relates to a drug response prediction or diagnostic kit comprising the composition.

The kit of the present invention is a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, which is a marker for predicting or diagnosing drug response, and is a nucleotide selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: (SNP) of the drug. ≪ / RTI > The drug response prediction or diagnostic kit of the present invention may include one or more other component compositions, solutions or devices suitable for the assay, as well as polynucleotides, primers or probes for identifying the SNP.

For example, the kit of the present invention may be a kit containing essential elements necessary for performing PCR. The PCR kit may contain test tubes or other appropriate containers, reaction buffers (varying in pH and magnesium concentration), deoxynucleotides (dNTPs), Taq polymerases and reverse transcriptase enzymes, as well as specific polynucleotides, primers or probes specific for the SNPs. , DNase, RNAse inhibitors, DEPC-water and sterile water, and the like.

In addition, the kit of the present invention may be a kit for predicting or diagnosing a drug response, which includes essential elements necessary for carrying out a DNA chip. The DNA chip kit may include a substrate to which a specific polynucleotide, a primer or a probe for the SNP is attached, and the substrate may include a nucleic acid corresponding to a quantitative control gene or a fragment thereof.

In another aspect, the present invention relates to a microarray for predicting or diagnosing a drug response comprising the composition.

The microarray may be composed of a conventional microarray except that it comprises the polynucleotide, primer or probe of the present invention.

The hybridization of nucleic acids on a microarray and the detection of hybridization results are well known in the art. The detection may be performed, for example, by labeling the nucleic acid sample with a labeling substance capable of generating a detectable signal including a fluorescent substance, such as Cy3 and Cy5, and then hybridizing on the microarray, The hybridization result can be detected.

In another aspect, the present invention provides a method for producing a sample, comprising: (a) separating a sample from a sample; And (b) identifying a base selected from the group consisting of 1207th, 1536th, 1594th and 2319th bases in the polynucleotide of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene in the separated sample, or Identifying an amino acid selected from the group consisting of 271, 381, 400, and 642-th SLCO2B1 proteins in the polypeptide consisting of SEQ ID NO: 2.

In the present invention, " specimen " means an individual to be examined, means the entire animal including a human, and preferably a human (subject).

The " sample " of the present invention is a concept including all of biological samples such as blood, skin, skin cells, mucosal cells and hair of a specimen.

In one embodiment of the present invention, when the 1207th base of SEQ ID NO: 1 as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene is replaced by thymine (T) in guanine (G), 271 of the polypeptide consisting of SEQ ID NO: 2 Th amino acid may be substituted with arginine to leucine. When these mutations are present, it can be judged that the drug absorption is promoted.

In another embodiment of the present invention, when the 1536th base of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene is replaced by thymine (T) in guanine (G) Lt; / RTI > amino acid may be substituted with valine to leucine. In the case of having such a mutation type, it can be judged that the drug absorption is inhibited.

In another embodiment of the present invention, when the 1594th base of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene is replaced by thymine (T) in guanine (G), the polypeptide of SEQ ID NO: 2 And the 400th amino acid may be substituted with leucine at arginine. When these mutations are present, it can be judged that the drug absorption is promoted.

In another embodiment of the present invention, when the 2319 base in SEQ ID NO: 1 is substituted with cytosine (C) in the thymine (T) as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the polypeptide of SEQ ID NO: 2 The 642nd amino acid may have a variant in which the histidine is substituted in the tyrosine. When these mutations are present, it can be judged that the drug absorption is promoted.

The drug of the present invention refers to all known drugs related to SLCO2B1, and more specifically, glibenclamide, estrone-3-sulfate, benzylpenicillin, fexofenadine, Bosentan, Statin, pravastatin, fluvastatin, atorvastatin, rosuvastatin, and the like, and the kind thereof is not particularly limited.

In the following examples of the present invention, the drug-transferring ability of each of four mutant SLCO2B1 was tested using estrone-3-sulfate as a representative drug transported by SLCO2B1 protein. As a result, R271L, R400L and Y642H of SLCO2B1 gene In the case of the mutant form, it was confirmed that more effective drug delivery was achieved than the normal drug delivery, and that the V381L mutant of the SLCO2B1 gene was not able to transport drugs normally. Therefore, in the case of a sample (subject) having the R271L, R400L and Y642H variants of the SLCO2B1 gene, the estrone-3-sulfate drug may be prescribed at a reduced level than the standard amount, or a specimen having the V381L mutant form of the SLCO2B1 gene The subject can provide information for drug selection decisions in a manner that does not prescribe estrone-3-sulfate drugs.

In another embodiment, the present invention provides a nucleic acid sample isolated from a specimen, wherein the SNP region of the SLCO2B1 gene is a polynucleotide consisting of the polynucleotide of SEQ ID NO: 1 and consisting of the 1207th, 1536th, 1594th and 2319th bases ≪ / RTI > Or identifying the amino acid selected from the group consisting of 271, 381, 400 and 642 in the polypeptide consisting of SEQ ID NO: 2 in the protein sample isolated from the sample. .

In one embodiment of the present invention, the mutated genotype of SLCO2B1 is a polynucleotide consisting of SEQ ID NO: 1, wherein the 1207th base is substituted with guanine (G) to thymine (T); The 1536th base is replaced by thymine (T) in guanine (G); The 1594th base is replaced with thymine (T) in guanine (G); And at least one mutation selected from the group wherein the 2319 base is substituted with cytosine (C) from thymine (T).

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1>

SLCO2B1  Four mutated genes found in genes

Genomic sequencing (NGS) analysis of more than 200 Korean specimens was carried out at the Genomic Center for Disease Control (National Institute for Disease Control and Prevention) and the Next Generation Custom Medical Genome Project. Genetic mutation genes were identified and four species of SLCO2B1 gene The mutation genes are shown below.

Four newly discovered mutant genes from the SLCO2B1 gene protein AA change SNP SLCO2B1 R271L 1207G> T SLCO2B1 V381L 1536G> T SLCO2B1 R400L 1594G > T SLCO2B1 Y642H 2319T> C

< Example  2>

SLCO2B1  Construction of expression vector for novel mutant gene

A total of four SLCO2B1 mutation genes were inserted into the pcDNA 3.1 expression vector (Promega) to produce four SLCO2B1 mutant gene expression vectors. For the amplification of SLCO2B1 gene, one of pcDNA3.1 (+) expression vector multicloning sites, EcoRI site, was constructed by attaching to the end of SLCO2B1 amplification specific primer. For each of the four variants, four expression vectors were prepared using QuikChangeXL Site-Directed Mutagenesis Kit (Agilent Technologies).

< Example  3>

Variant SLCO2B1 of Drug delivery ability  Measure

In this experiment, the expression vector containing the SLCO2B1 novel mutant gene prepared in Example 2 was transfected into HEK293 (human embryonic kidney) cell line, and the drug of the SLCO2B1 protein in the cell line into which the mutant SLCO2B1 gene was introduced The ability to carry out was performed using drugs with radioactive isotopes.

In detail, HEK293 cell line (1.3x10 ⁵ cells / well) was cultured in a single cell layer in a 24 well plate coated with poly D-lysine, and then cultured in Lipofectamine 2000 (Invitrogen) Were transfected into HEK293 cell line and expressed at 37 DEG C for 24 hours. The transfected HEK293 cell line was washed twice with DPBS (Dulbecco's Phosphate Buffered Saline) and then pre-warmed with DPBS (500 μl / well) in a constant temperature bath at 37 ° C for 10 minutes. After that, DPBS was removed and about 22 nM of estrone labeled with tritium ([ ³ H]) (Estrone sulfate, ARC (American Radio Label Chemicals, Inc.)) was reacted in a constant temperature water bath at 37 ° C for 5 minutes. At this time, 100 μM of sulfobromophthalein (BSP, Sigma Aldrich) was used as a transport inhibitor of SLCO2B1. After the reaction, the cells were washed 3 times with cold DPBS, and the HEK293 cell line was lysed with 200 μl / well of 0.1 N NaOH for 1 hour. The solution was transferred to a 24-well flexible microplate (PerkinElmer), reacted with a supermix (PerkinElmer) 500 μl / well in a shaker at room temperature for 4 hours, and then radioactively analyzed with a Beta counter.

- dpm: the number of radioactive elements per minute

- substrate reaction rate = pmol / mg protein / min

- Maximum reaction rate (Vmax): At a certain point in time, the formation rate of the product is not increased even if the substrate concentration is increased

- Michaelis lieutenant constant (Km): a parameter indicating the affinity of the enzyme with the substrate

Results of radioisotope analysis (unit dpm) pcDNA WT R271L V381L R400L Y642H ES 141 595 611 382 629 711 ES + BSP 113 112 156 137 130 140

As a result, as shown in Table 2 and FIG. 2, it was confirmed that the number of radioactive element decay was slightly increased by the R271L, R400L and Y642H mutations of the SLCO2B1 gene, whereas the number of radioactive element decay by the V381L mutation was about two times Respectively. This indicates that the transporter drug transport capacity is inhibited by the SLCO2B1 V381L gene mutation.

< Example  4>

Variant SLCO2B1 Pharmacokinetic analysis of

In this experiment, for the pharmacokinetic analysis of the mutant SLCO2B1 protein according to the substrate concentration, estrone sulfate labeled with tritium ([ ³ H]) was treated at 5, 10, 30, 50, 3> were performed under the same conditions.

In detail, wild-type and mutant SLCO2B1-transfected HEK293 cells were washed twice with DPBS (Dulbecco's Phosphate Buffered Saline) and then pre-warmed with DPBS (500 μl / well) in a constant temperature bath at 37 ° C for 10 minutes. Then, DPBS was removed, and estrone sulfate labeled with tritium ([ ³ H]) was treated at 5, 10, 30, 50, and 100 μM, respectively, and reacted in a constant temperature water bath at 37 ° C for 5 minutes. Experiments were conducted in the same manner as in Example 3 described below.

Enzymatic analysis of wild-type and mutant SLCO2B1 proteins The maximum reaction rate (Vmax) and enzyme-substrate affinity (Km) Variant Km (? M) Vmax (pmol / mg protein / min) WT (wild type) 23.5 10.3 R271L 58 16.4 V381L 26.0 5.8 R400L 39.3 15.7 Y642H 43.7 18.7

As a result, as shown in Table 3 and FIG. 3, the R271L, R400L and Y642H variants of the SLCO2B1 gene showed more effective drug transport than the normal drug transport, and the SLC2B1 V381L variant showed normal drug transport I can see that I can not. This means that the R271L, R400L and Y642H variants of the SLCO2B1 gene promotes the function of the carrier, while the SLC2B1 V381L variant can result in the inhibition of the enzyme activity due to the inability of the carrier to function.

< Example  5>

Confirmation of protein expression measurement

Western blotting was performed to confirm that the mutant SLCO2B1 protein was normally expressed by intracellular transfection of the expression vector prepared in Example 2 above.

Specifically, 20 μg of each sample of the lysate of cells transfected with the above-described expression vector was boiled for 5 minutes at 100 ° C. together with 4 × sample loading buffer solution (NuPAGE), and centrifuged at 12,000 rpm for 1 minute And prepared separately. The prepared samples were loaded on a 12% polyacrylamide gel and developed for 2 hours at 120V. The electrophoresed proteins were transferred to a PVDF membrane and immunoblotted with SLCO2B1 polyclonal antibody (SantaCruz) and goat anti-rabbit IgG (goat anti-rabbit IgG). Thereafter, a luminescence reaction was performed using an ECL (Enhanced chemiluminescent) solution (Millipore) and sensitized with Image J to confirm protein expression.

As a result, as shown in Fig. 4, it was confirmed that the difference in the drug-transporting ability of the mutant SLCO2B1 protein was due to the difference in the amount of protein.

< Example  6>

SLCO2B1 Membrane potential  Pattern analysis of proteins

In this experiment, the pattern of SLCO2B1 protein was analyzed using TOPO2 (Transmembrane Protein Display) to determine the mutation position of the SLCO2B1 gene. According to NCBI, the total membrane potential of the SLCO2B1 protein is 12, and translation is known to start in the cytoplasm.

As a result, as shown in Fig. 5, it was confirmed that the mutated SLCO2B1 R271L, R400L and Y642H were located outside the cell, and the mutant SLCO2B1 V381L was located inside the membrane.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

SLCO2B1: Solute carrier organic anion transporter family member 2B1
SNP: single nucleotide polymorphism
ES: Estrone-3-sulfate
BSP: Bromsulphthalein (BSP)

<110> inje university industry-academic cooperation foundation <120> Novel SLCO2B1 Single Nucleotide Polymorphism Markers and Uses          Thereof <130> PN1502-036 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 4362 <212> DNA <213> SLCO2B1 polynucleotide sequence <400> 1 aaaaggacag gctgaggctg agcctcaggg aggcggggtg cagcttcctc ttcacagaga 60 ggctggactt gaacaaaccc agccttctat acaaccgtgg agccagggca agggagagac 120 agaaggagca agtgacccag ggagacaaac acttggagat acttggggct gagtttgagc 180 aagactccct aacctgtgtc tggacaagtc tgatgtcctg tgtggcccaa gaagaactga 240 ccccgtgtct ggagctccca ccgttattgc atccctgctg tggctcacct gctgctgtct 300 ccaggagccc ctgagaagat ttgcttcctc tcccctgcta agctccaggt cctgagatta 360 aattaggggc tggagctcac tgcactccag cagtcatggg acccaggata gggccagcgg 420 gtgaggtacc ccaggtacca gacaaggaaa ccaaagccac aatgggcaca gaaaacacac 480 ctggaggcaa agccagccca gaccctcagg acgtgcggcc aagtgtgttc cataacatca 540 agctgttcgt tctgtgccac agcctgctgc agctggcgca gctcatgatc tccggctacc 600 taaagagctc catctccaca gtggagaagc gcttcggcct ctccagccag acgtcggggc 660 tgctggcctc cttcaacgag gtggggaaca cagccttgat tgtgtttgtg agctattttg 720 gcagccgggt gcaccgaccc cgaatgattg gctatggggc tatccttgtg gccctggcgg 780 gcctgctcat gactctcccg cacttcatct cggagccata ccgctacgac aacaccagcc 840 ctgaggatat gccacaggac ttcaaggctt ccctgtgcct gcccacaacc tcggccccag 900 cctcggcccc ctccaatggc aactgctcaa gctacacaga aacccagcat ctgagtgtgg 960 tggggatcat gttcgtggca cagaccctgc tgggcgtggg cggggtgccc attcagccct 1020 ttggcatctc ctacatcgat gactttgccc acaacagcaa ctcgcccctc tacctcggga 1080 tcctgtttgc agtgaccatg atggggccag gcctggcctt tgggctgggc agcctcatgc 1140 tgcgccttta tgtggacatt aaccagatgc cagaaggtgg tatcagcctg accataaagg 1200 acccccgatg ggtgggtgcc tggtggctgg gtttcctcat cgctgccggt gcagtggccc 1260 tggctgccat cccctacttc ttcttcccca aggaaatgcc caaggaaaaa cgtgagcttc 1320 agtttcggcg aaaggtctta gcagtcacag actcacctgc caggaagggc aaggactctc 1380 cctctaagca gagccctggg gagtccacga agaagcagga tggcctagtc cagattgcac 1440 caaacctgac tgtgatccag ttcattaaag tcttccccag ggtgctgctg cagaccctac 1500 gccaccccat cttcctgctg gtggtcctgt cccaggtatg cttgtcatcc atggctgcgg 1560 gcatggccac cttcctgccc aagttcctgg agcgccagtt ttccatcaca gcctcctacg 1620 ccaacctgct catcggctgc ctctccttcc cttcggtcat cgtgggcatc gtggtgggtg 1680 gcgtcctggt caagcggctc cacctgggcc ctgtgggatg cggtgccctt tgcctgctgg 1740 ggatgctgct gtgcctcttc ttcagcctgc cgctcttctt tatcggctgc tccagccacc 1800 agattgcggg catcacacac cagaccagtg cccaccctgg gctggagctg tctccaagct 1860 gcatggaggc ctgctcctgc ccattggacg gctttaaccc tgtctgcgac cccagcactc 1920 gtgtggaata catcacaccc tgccacgcag gctgctcaag ctgggtggtc caggatgctc 1980 tggacaacag ccaggttttc tacaccaact gcagctgcgt ggtggagggc aaccccgtgc 2040 tggcaggatc ctgcgactca acgtgcagcc atctggtggt gcccttcctg ctcctggtca 2100 gcctgggctc ggccctggcc tgtctcaccc acacaccctc cttcatgctc atcctaagag 2160 gagtgaagaa agaagacaag actttggctg tgggcatcca gttcatgttc ctgaggattt 2220 tggcctggat gcccagcccc gtgatccacg gcagcgccat cgacaccacc tgtgtgcact 2280 gggccctgag ctgtgggcgt cgagctgtct gtcgctacta caataatgac ctgctccgaa 2340 accggttcat cggcctccag ttcttcttca aaacaggttc tgtgatctgc ttcgccttag 2400 ttttggctgt cctgaggcag caggacaaag aggcaaggac caaagagagc agatccagcc 2460 ctgccgtaga gcagcaattg ctagtgtcgg ggccagggaa gaagccagag gattcccgag 2520 tgtgagctgt cttggggccc cacctggcca agagtagcag ccacagcagt acctcctctg 2580 agtcctttgc ccaagattgg gtgtcaagag ccctgtgttc cattctggct cctccactaa 2640 attgctgtgt gacttcaggc aagacattga tcctctctca gcctttgctt gctagtctga 2700 accaaagagt tgtttgggca tttgctgtgt tggccatttc tggagcaaga gggtcttctt 2760 cctccttccc ccagccagcc agctgtcctg gggccaggct ttcctgggtg gaaagaagta 2820 tacctttccc tggggcccta ggatagcaaa gtgagccata gtgggccagg ctgccctcca 2880 tgctgggccc cagcccaggt ctgcactcgc ctggatcacc ttctttgagc cttagccatc 2940 tcctgtcagg taggaatgaa cttgccagcc ttcaggctcg ttcagctatg accatctgtg 3000 cggtcagggt acactcagct ctcctcccca actccagcag cctttaagaa gtgtcccttt 3060 ggcgccccct ggaggcagag cactgagctg gaccctgggt agactcccac agggaggacg 3120 gagctggcct caggagtggg acacccagac ttggcagggc cttcaagagg cctgtgtggg 3180 ggccccagga atccttagct gaagcgggga gactcactct ccatctcagg aaattctagc 3240 ccttgccctc agggagccac ggttgagggt gaggcccaac acctgcctta gggccctggg 3300 tgggcaagtc tgggccctgg ggtagggagg gagactcagg cccacacttg ggtattttct 3360 aattacagac aaacacacac tcagcgcgca ctcactgatt cctacacatt gccaagattt 3420 cacacatgtg accaggggcc accaaagtcc ctgtgacctt tgtgactagg atcctaattt 3480 ctctattttc tcctgggtgc ctgggtctgt gtcacctggg gcagtgtgga taatgtttag 3540 ttctgtgaca ctgttttttg ggggtggcac ctggttctcc gatgcctggg ctggtgtcag 3600 gcccaggact gtagtgctgg gagcagtaaa gctcagctct gtgtaatgag tgatgctatg 3660 gcttgctcgt gtcttatgat ccaatccttt tctacatcag cccttgtttt gttttatggc 3720 tagtcttatc tggcctggtt atttccttgc ggggaggaga gggtttgcta atctgctccc 3780 agcccaacct attaccaccc cacctcgctg ggacctactg ctcgggaggc agcagacagg 3840 gagccaccag cagtggcttc ctggccctgt gctgggggtg gggggaagct gggggcacat 3900 gtggcccttg ccttctgagc agctcccagt gccagggctt tgagactttc ccacatgata 3960 aaagaaaagg gaggtacaga agttccaatt ccctttttat tttgctggtt ggtatctgta 4020 aatgtttaat aaatatctga gcatgtatct atcaacgcca agaatttcaa agtctccttc 4080 aacaatatga ggcttttagg atgtttatat tccttcatcc ctcttgtttc ccaggttttg 4140 cagggaaaaa aagtctggaa ttatagatac agcttattat taaatttgtt cttgcataat 4200 gtctcttcta ttacaaaaat tctttcttca taaactgcat tagaggtttg caacaaccac 4260 atcatttcca ttaacttaga tttaggtttt actggattca ttgctcacca ttattgcttg 4320 tatattacat cttttccaat ctttaaaaaa aaaaaaaaaaaa 4362 <210> 2 <211> 709 <212> PRT <213> SLCO2B1 polynucleotide sequence <400> 2 Met Gly Pro Arg Ile Gly Pro Ala Gly Glu Val Pro Gln Val Pro Asp   1 5 10 15 Lys Glu Thr Lys Ala Thr Met Gly Thr Glu Asn Thr Pro Gly Gly Lys              20 25 30 Ala Ser Pro Asp Pro Gln Asp Val Arg Pro Ser Val Phe His Asn Ile          35 40 45 Lys Leu Phe Val Leu Cys His Ser Leu Leu Gln Leu Ala Gln Leu Met      50 55 60 Ile Ser Gly Tyr Leu Lys Ser Ser Ile Ser Thr Val Glu Lys Arg Phe  65 70 75 80 Gly Leu Ser Ser Gln Thr Ser Gly Leu Leu Ala Ser Phe Asn Glu Val                  85 90 95 Gly Asn Thr Ala Leu Ile Val Phe Val Ser Tyr Phe Gly Ser Arg Val             100 105 110 His Arg Pro Arg Met Ile Gly Tyr Gly Ala Ile Leu Val Ala Leu Ala         115 120 125 Gly Leu Leu Met Thr Leu Pro His Phe Ile Ser Glu Pro Tyr Arg Tyr     130 135 140 Asp Asn Thr Ser Pro Glu Asp Met Pro Gln Asp Phe Lys Ala Ser Leu 145 150 155 160 Cys Leu Pro Thr Thr Ser Pro Ala Ser Ala Pro Ser Asn Gly Asn                 165 170 175 Cys Ser Ser Tyr Thr Glu Thr Gln His Leu Ser Val Val Gly Ile Met             180 185 190 Phe Val Ala Gln Thr Leu Leu Gly Val Gly Gly Val Pro Ile Gln Pro         195 200 205 Phe Gly Ile Ser Tyr Ile Asp Asp Phe Ala His His Ser Asn Ser Pro     210 215 220 Leu Tyr Leu Gly Ile Leu Phe Ala Val Thr Met Met Gly Pro Gly Leu 225 230 235 240 Ala Phe Gly Leu Gly Ser Leu Met Leu Arg Leu Tyr Val Asp Ile Asn                 245 250 255 Gln Met Pro Glu Gly Gly Ile Ser Leu Thr Ile Lys Asp Pro Arg Trp             260 265 270 Val Gly Ala Trp Trp Leu Gly Phe Leu Ile Ala Ala Gly Ala Val Ala         275 280 285 Leu Ala Ala Ile Pro Tyr Phe Phe Phe Pro Lys Glu Met Pro Lys Glu     290 295 300 Lys Arg Glu Leu Gln Phe Arg Arg Lys Val Leu Ala Val Thr Asp Ser 305 310 315 320 Pro Ala Arg Lys Gly Lys Asp Ser Pro Ser Lys Gln Ser Pro Gly Glu                 325 330 335 Ser Thr Lys Lys Gln Asp Gly Leu Val Gln Ile Ala Pro Asn Leu Thr             340 345 350 Val Ile Gln Phe Ile Lys Val Phe Pro Arg Val Leu Leu Gln Thr Leu         355 360 365 Arg His Pro Ile Phe Leu Leu Val Val Leu Ser Gln Val Cys Leu Ser     370 375 380 Ser Met Ala Ala Gly Met Ala Thr Phe Leu Pro Lys Phe Leu Glu Arg 385 390 395 400 Gln Phe Ser Ile Thr Ala Ser Tyr Ala Asn Leu Leu Ile Gly Cys Leu                 405 410 415 Ser Phe Pro Ser Val Ile Val Gly Ile Val Val Gly Gly Val Leu Val             420 425 430 Lys Arg Leu His Leu Gly Pro Val Gly Cys Gly Ala Leu Cys Leu Leu         435 440 445 Gly Met Leu Leu Cys Leu Phe Phe Ser Leu Pro Leu Phe Phe Ile Gly     450 455 460 Cys Ser Ser His Gln Ile Ala Gly Ile Thr His Gln Thr Ser Ala His 465 470 475 480 Pro Gly Leu Glu Leu Ser Pro Ser Cys Met Glu Ala Cys Ser Cys Pro                 485 490 495 Leu Asp Gly Phe Asn Pro Val Cys Asp Pro Ser Thr Arg Val Glu Tyr             500 505 510 Ile Thr Pro Cys His Ala Gly Cys Ser Ser Trp Val Val Gln Asp Ala         515 520 525 Leu Asp Asn Ser Gln Val Phe Tyr Thr Asn Cys Ser Cys Val Val Glu     530 535 540 Gly Asn Pro Val Leu Ala Gly Ser Cys Asp Ser Thr Cys Ser His Leu 545 550 555 560 Val Val Pro Phe Leu Leu Leu Val Ser Leu Gly Ser Ala Leu Ala Cys                 565 570 575 Leu Thr His Thr Pro Ser Phe Met Leu Ile Leu Arg Gly Val Lys Lys             580 585 590 Glu Asp Lys Thr Leu Ala Val Gly Ile Gln Phe Met Phe Leu Arg Ile         595 600 605 Leu Ala Trp Met Pro Ser Pro Val Ile His Gly Ser Ala Ile Asp Thr     610 615 620 Thr Cys Val His Trp Ala Leu Ser Cys Gly Arg Arg Ala Val Cys Arg 625 630 635 640 Tyr Tyr Asn Asn Asp Leu Leu Arg Asn Arg Phe Ile Gly Leu Gln Phe                 645 650 655 Phe Phe Lys Thr Gly Ser Val Ile Cys Phe Ala Leu Val Leu Ala Val             660 665 670 Leu Arg Gln Gln Asp Lys Glu Ala Arg Thr Lys Glu Ser Arg Ser Ser         675 680 685 Pro Ala Val Glu Gln Gln Leu Leu Val Ser Gly Pro Gly Lys Lys Pro     690 695 700 Glu Asp Ser Arg Val 705

Claims (20)

(SNP) of the base selected from the group consisting of 1207th, 1536th, 1594th and 2319th bases in the polynucleotide of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene ; Or an amino acid selected from the group consisting of 271st, 381st, 400th and 642nd in the polypeptide consisting of SEQ ID NO: 2, wherein the drug is an estrone- 3-sulfate. &Lt; / RTI &gt; The method according to claim 1,
The 1207th base is G or T; The 1536th base is G or T; The 1594th base is G or T; Or the 2319 base is T or &lt; RTI ID = 0.0 &gt; C. &lt; / RTI &gt; delete The method according to claim 1,
The agent capable of detecting the single nucleotide polymorphism (SNP) is a polynucleotide comprising 10 to 100 nucleotide polymorphisms (SNPs) including SNPs selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: Wherein the primer is a primer capable of amplifying a polynucleotide composed of a continuous base or a complementary polynucleotide thereof. The method according to claim 1,
The agent capable of detecting the single nucleotide polymorphism (SNP) is a polynucleotide comprising 10 to 100 nucleotide polymorphisms (SNPs) including SNPs selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: Wherein the probe is a probe capable of hybridizing with a polynucleotide composed of a continuous base. The method according to claim 1,
Wherein the agent capable of detecting the amino acid is an antibody capable of forming a bond with an amino acid selected from the group consisting of 271st, 381st, 400th and 642nd in the polypeptide consisting of SEQ ID NO: 2. Or diagnostic composition. 8. A kit for predicting or diagnosing a drug response comprising a composition according to any one of claims 1, 2 and 4 to 6, wherein the drug is estrone. (a) separating a sample from a specimen; And
(b) identifying a base selected from the group consisting of the 1207th, 1536th, 1594th and 2319th bases in the polynucleotide of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene in the separated sample, Identifying an amino acid selected from the group consisting of the 271st, 381st, 400th and 642nd of the SLCO2B1 protein in the polypeptide consisting of SEQ ID NO: 2, wherein the drug is an estrone How it is. 9. The method of claim 8,
When the 1207th base of SEQ ID NO: 1 as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene is replaced with thymine (T) in guanine (G), the 271st amino acid of the polypeptide of SEQ ID NO: 2 is substituted with leucine in arginine Wherein the drug is selected from the group consisting of: &lt; RTI ID = 0.0 &gt; a &lt; / RTI &gt; 9. The method of claim 8,
When the 1536th base in SEQ ID NO: 1 is substituted with thymine (T) in guanine (G) as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the 381st amino acid of the polypeptide consisting of SEQ ID NO: 2 is substituted with valine to leucine Wherein the drug is selected from the group consisting of: &lt; RTI ID = 0.0 &gt; a &lt; / RTI &gt; 9. The method of claim 8,
When the 1594th base of SEQ ID NO: 1 is substituted with thymine (T) in guanine (G) as the single nucleotide polymorphism (SNP) region of the SLCO2B1 gene, the 400th amino acid of the polypeptide consisting of SEQ ID NO: 2 is substituted with arginine to leucine Wherein the drug is selected from the group consisting of: &lt; RTI ID = 0.0 &gt; a &lt; / RTI &gt; 9. The method of claim 8,
When the 2319th base in SEQ ID NO: 1 is substituted with cytosine (C) in the thymine (T) as the single base polymorphism (SNP) region of the SLCO2B1 gene, the 642nd amino acid of the polypeptide consisting of SEQ ID NO: 2 is replaced with histidine in the tyrosine Wherein the drug is selected from the group consisting of: &lt; RTI ID = 0.0 &gt; a &lt; / RTI &gt; 11. The method of claim 10,
And when it has the above-mentioned substituted mutation type, it is determined that the drug absorption is inhibited. The method according to claim 9, 11, or 12,
And when the drug has the substituted mutation type, it is determined that the drug absorption is promoted. delete 11. The method of claim 10,
As a result of inhibition of drug absorption on estrone-3-sulfate drugs when the above-mentioned substituted mutant forms are inhibited, estrone-3-sulfate drug is not prescribed in the case of a sample having such a substituted mutant form Characterized in that the method comprises: Identifying a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 in the polynucleotide of SEQ ID NO: 1 as a single nucleotide polymorphism (SNP) region of the SLCO2B1 gene in a nucleic acid sample isolated from a specimen; Or identifying the amino acid selected from the group consisting of 271, 381, 400 and 642 in the polypeptide consisting of SEQ ID NO: 2 in the protein sample isolated from the sample. . 18. The method of claim 17,
The mutated genotype of SLCO2B1 is a polynucleotide consisting of SEQ ID NO: 1 in which the 1207th base is substituted with guanine (G) to thymine (T); The 1536th base is replaced by thymine (T) in guanine (G); The 1594th base is replaced with thymine (T) in guanine (G); And the 2319 base is substituted by cytosine (C) in thymine (T). Identifying a base selected from the group consisting of nucleotides 1207, 1536, 1594 and 2319 of the SLCO2B1 gene of SEQ ID NO: 1. 20. The method of claim 19,
The method comprises determining whether the 1207th base of the SLCO2B1 gene in the polynucleotide of SEQ ID NO: 1 is substituted with guanine (G) to thymine (T); Whether or not the 1536th base of the SLCO2B1 gene of SEQ ID NO: 1 is replaced with thymine (T) in guanine (G); Whether the 1594th base of the SLCO2B1 gene of SEQ ID NO: 1 was substituted with thymine (T) in guanine (G); Or confirming whether the 2319 base of the SLCO2B1 gene of SEQ ID NO: 1 has been substituted with cytosine (C) in thymine (T).

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