JP2012070732A - Method of evaluating drug sensitivity and illness vulnerability through adrenergic receptor gene analysis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method of evaluating (predicting, etc.) individual differences (tendency of each individual) in drug sensitivity or illness vulnerability by using adrenergic receptor gene polymorphism, etc.SOLUTION: The method of evaluating drug sensitivity or the method of evaluating illness vulnerability comprises associating the gene polymorphism of an adrenergic receptor gene or a haplotype comprising the gene polymorphism with an individual's drug sensitivity or illness vulnerability.

Description

  The present invention relates to a method for evaluating drug sensitivity and disease vulnerability by analyzing an adrenergic receptor (ADR) gene. Specifically, the ADR gene polymorphism or a haplotype composed of the gene polymorphism is associated with drug sensitivity and disease vulnerability of an individual, more specifically, a method for evaluating drug sensitivity and disease vulnerability, etc. The present invention relates to a method for evaluating the tendency of drug sensitivity and disease vulnerability in an individual based on the analysis result of the gene polymorphism or haplotype.

  Pain is the most frequently observed medical condition, and pain associated with the disease is often more serious for the patient than the disease itself. Although the pain sensation has an important role in that it is a living body warning system, if the excessive pain is not properly controlled, the quality of life (QOL) is significantly lowered. In recent years, the importance of pain control has been recognized, and palliative medicine including pain treatment has been making dramatic progress, and the use opportunities and usage of various analgesics are increasing.

So far, analgesic effects via an adrenergic receptor have been confirmed, such as an analgesic being obtained by administering an α ₂ adrenergic receptor agonist to the human spinal subarachnoid space (Non-patent Document 1). In addition, it has been reported that esmolol, an ultra-short-acting β ₁ adrenergic receptor blocker, suppresses pain responses caused by formalin in animal experiments (Non-patent Document 2). In addition, serotonin and noradrenaline reuptake inhibitor (SNRI) duloxetine is the first in the United States to treat neuropathic pain caused by diabetic neuropathy, which has been suggested to be caused by functional disruption of the descending inhibitory system. Approved (Non-Patent Document 3). In addition, administration of α ₂ receptor agonists or β receptor blockers to the amygdala striatal nucleus in rats alleviates conditioned place aversion (CPA) caused by opioid withdrawal symptoms and is caused by formalin It has been reported that place aversive reactions are almost completely suppressed by administering morphine to the bilateral basolateral nucleus of the amygdala 5 minutes before subcutaneous administration of formalin. It has also been suggested that both adrenergic receptors and opioids are involved (Non-Patent Documents 4 and 5).

Gordh T Jr. et al., Effect of epidural clonidine on spinal cord blood flow and regional and central hemodynamics in pigs, Anesth. Analg., (1986) 65: 1312-1318. Davidson EM. Et al., Antinociceptive and cardiovascular properties of esmolol following formalin injection in rats, Can. J. Anaesth., (2001) 48: 59-64. Ikeda, K. et al., Review of duloxetine in the management of diabetic peripheral neuropathic pain, Vasc.Health Risk Manag., (2007) 3: 833-844. Delfs JM, et al., Noradrenaline in the ventral forebrain is critical for opiate withdrawal-induced aversion., Nature, (2000) 403: 430-434. Deyama, S. et al., Inhibition of glutamatergic transmission by morphine in the basolateral amygdaloid nucleus reduces pain-induced aversion., Neurosci. Res., (2007) 59: 199-204.

  The problems to be solved by the present invention include drug sensitivity or disease vulnerability, specifically, the number of analgesic administration necessary times, total analgesic amount, pain sensitivity, drug dependence vulnerability, and prolonged psychostimulant psychosis Another object is to provide a method for evaluating (predicting) an individual difference (trend for each individual) of disease vulnerability using an adrenergic receptor gene polymorphism or the like.

  The present inventor paid attention to an adrenergic receptor gene and conducted earnest research based on conventional knowledge and clinical data. As a result, several useful gene polymorphisms were identified by analyzing the correlation between various adrenergic receptor gene polymorphisms and drug susceptibility such as analgesics or disease vulnerability including pain sensitivity. They found linkage disequilibrium between these identified polymorphisms, and revealed a significant correlation with drug sensitivity or disease vulnerability.

  Specifically, it has been clarified that the required amount of analgesics changes and the threshold of pain sensitivity changes due to differences in specific adrenergic receptor gene polymorphisms, and the present invention has been completed.

That is, the present invention is as follows.
1. A method for evaluating drug sensitivity, comprising associating a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism with an individual drug sensitivity.
Examples of the evaluation method include a method characterized by evaluating the tendency of an individual to have drug sensitivity based on the analysis result of the gene polymorphism or the haplotype.
2. The following steps:
(1) A step of performing linkage disequilibrium analysis and haplotype analysis in a healthy person, and selecting a gene polymorphism in the linkage disequilibrium block;
(2) analyzing the association between the genotype of the gene polymorphism in the subject and drug sensitivity, and (3) using the gene polymorphism significantly associated with drug sensitivity in the subject for evaluating drug sensitivity. The method according to item 1 above.
3. A method for evaluating disease vulnerability, comprising associating a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism with a disease vulnerability of an individual.
Examples of the evaluation method include a method characterized by evaluating the tendency of an individual for disease vulnerability based on the analysis result of the genetic polymorphism or the haplotype.
4). The following steps:
(1) A step of performing linkage disequilibrium analysis and haplotype analysis in a healthy person, and selecting a gene polymorphism in the linkage disequilibrium block;
(2) Analyzing the association between the genotype of the gene polymorphism in the subject and pain sensitivity, and (3) Using the gene polymorphism significantly associated with pain sensitivity in the subject for evaluating disease vulnerability The method according to item 3 above, comprising:
5. 5. The method according to item 3 or 4 above, wherein the disease vulnerability is drug sensitivity / pain sensitivity or vulnerability to drug dependence.
6). The genetic polymorphism is at least one selected from the group consisting of a single nucleotide polymorphism, an insertion polymorphism, a deletion polymorphism, and a nucleotide repeat polymorphism, according to any one of the preceding items 1 to 5. Method.
7). The gene polymorphism is ADRB2 gene (ADRB2 gene which is a subtype of the adrenergic receptor gene (hereinafter the same)) rs888956, rs10075525, rs2116719, rs10059242, rs11742884, rs2116713, rs2400707, rs1042713, rs1042717, rs4705271, rs3857420, rs6888011, rs11959113 , Rs17707884, rs11742519, rs10214302, rs11168071, rs11640705, rs1432631, rs919725, rs1864932, rs11740830, rs4705283 and rs1181135, ADRB1 subtype gene (adrenergic receptor gene subtype ADRB1 gene (same below)) rs1801252, rs1801253, rs1801253, 14 , Rs6585258, rs2480792, rs11196589, rs623499, rs2782979, rs2782980, rs12771397, rs11196592, rs11196597, rs10787516 and rs4918887, and the ADRA2A subtype polymorphisms rs990428, rs11195417, rs7074629, rs491589, rs34665721, rs745557, 4579013, rs745557, , Rs1343450, rs1556716, rs7897445, rs1537770, rs2032023, rs17128431, rs706956 7. The method according to any one of items 1 to 6, which is at least one selected from 4, rs954385, rs1889745, rs1335706, rs1335704, rs1335703, rs11195442, rs12220858, rs4545476, rs7908674, rs10885102, rs7084236, rs7088234 and rs7079973.
8). 8. The method according to any one of items 1 to 7, wherein the haplotype is at least one selected from those shown in the following table.

9. 9. A method for determining the type, amount and / or number of administrations of a drug to be administered to an individual using the result evaluated by the method according to any one of items 1 to 8 as an index.
10. 10. A method for predicting side effects of a drug administered to an individual using the result evaluated by the method according to any one of items 1 to 9 as an index.
11. 11. The method according to any one of the aforementioned items 1, 2, 5, 9, and 10, wherein the drug is an opioid receptor function modifier and / or an adrenergic receptor function modifier.
12 Opioid receptor function modifiers include methamphetamine, methylenedioxymethamphetamine, amphetamine, dextroamphetamine, dopamine, morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, nalolphine, levalorphan Pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, laughter, F3 (1-chloro- 1,2,2trifluorocyclobutane), halocene, estradiol, dithiothreitol, At least one selected from the group consisting of thioridazine, pimozide, fluoxetine, paroxetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodir, bupivacaine, terthiapine, clozapine, haloperidol, SCH23390 and cocaine; Receptor function modifiers are adrenaline, noradrenaline, dopamine, phenylephrine, naphazoline, α-methylnoradrenaline, oxymetazoline, xylometazoline, clonidine, guanfacine, guanabenz, guanoxabenz, guanethidine, xylazine, methyldopa, anthromidamine, amidazoline, amidazoline, amidazoline Apraclonidine, brimonidine, silazoline, detomidine, de Cosmedetomidine, epinephrine, ethylephrine, indanidine, lofexidine, medetomidine, mephentermine, metallaminol, methoxamine, middolin, mibazelol, norphenephrine, octopamine, oxymetazoline, phenylpropanolamine, rilmenidine, romifidine, cinefelidine, tinipedamine , Ergometrine, phentolamine, trazoline, bunazosin, naphthopidyl, prazosin, terazosin, doxazosin, tamsulosin, silodosin, alfuzosin, trimazosin, phenoxybenzamine, amitriptyline, clomipramine, doxepin, trimipramine, α-methyldopa, yoxinol , Dobutamine, isoproterenol, xamoterol, epinephrine, isoprenaline, salbutamol (albuterol), levosalbutamol (levalbuterol), terbutaline, metaproterenol, fenoterol, bitolterol, pyrbuterol, procaterol, salmeterol, formoterol, bambuterol, clenbuterol Isoethalin, ritodrine, vitorterol mesylate, L-796568, amibegron, sorabegron, albutamine, befnolol, bromoacetylalprenololmenthane, broxaterol, cimaterol, silazoline, denopamine, dopexamine, ethylephrine, hexoprenalin, isopenaminteline, purigenamine , Nilidrin, oxyfedrine, Renalterol, ractopamine, reproterol, limiterol, tretoquinol, tulobuterol, zilpaterol, ginterol, alprenolol, bucindolol, sotalol, bopindolol, pindolol, timolol, dichloroisoprenalin, alprenolol, carteolol, indenolol, bunitrolol, penbutrolol, penbutrolol , Nipradilol, chirisolol, acebutolol, seriprolol, metoprolol, atenolol, bisoprolol, betaxolol, practolol, bevantolol, esmolol, nebivolol, butoxamine, ICI-118,551, SR 59230A, mepindolol, oxprenolol, landiolol, levobrolol Roll, labetalol, cal Vezirol, arotinolol, amosulalol, amphetamine, droxidopa, tyramine, ephedrine, pseudoephedrine, atomoxetine (tomoxitin), mazindol, reboxetine, viloxazine, amineptine, bupropion, dexmethylphenidate, fencampamine, fencamamine, fetadamine, fetamine Prolintan, pyrovalerone, diphemetrex, desvenlafaxine, duloxetine, milnacipran, venlafaxine, buttriptyline, desipramine, dosrepin, imipramine, lofepramine, nortriptyline, protriptyline, amoxapine, maprotiline, mianserin, cyclobenzaprine , Mesocarb, nefadozon, nehopam, sibutra , Tapentadol, tramadol, ziprasidone, adhiperifolin, hyperforin, cocaine, desoxypipradorol, diphenylprolinol, tylenedioxypylobalerone, cycladine dol, esreboxetine, manifaxin, nisoxetine, ladafaxin, tandamine, WYE- 103231, 1- (Indolin-1-yl) -1-phenyl-3-propan-2-olamines, 1- or 3- (3-Amino-2-hydroxy-1-phenyl propyl) -1,3-dihydro- 2H-benzimidazol-2-ones, (+)-S-21, thienyl-based, bicifazine, brasofensin, diclofensin, DOV-21,947, DOV-102,677, DOV-216,303, indatraline, NS-2359, oxapro 12. The method according to item 11 above, which is at least one selected from the group consisting of tillin, SEP-225,289, SEP-227,162, tesofensin, and Dutchville.
13. At least the 51st base in the base sequence shown in any one of SEQ ID NOs: 1-38 and 47-76, which can specifically hybridize to a DNA fragment containing a gene polymorphism of an adrenergic receptor gene 13. The method according to any one of items 1 to 12, wherein an oligonucleotide having a base sequence having a length of 10 bases or a base sequence complementary to the base sequence is used.
14 14. The method according to item 13 above, wherein the oligonucleotide has a length of 10 to 150 bases.
15. In the preceding item 13 or 14, wherein the oligonucleotide is an oligonucleotide selected from the group consisting of the base sequence shown in any one of SEQ ID NOs: 1-38 and 47-76 or a base sequence complementary to the base sequence The method described.
20. The primer which consists of a base sequence shown by any one of sequence number 39-46 used for the detection of the adrenergic receptor gene polymorphism in the method of any one of the preceding clauses 1-15.
21. 21. A drug sensitivity evaluation kit comprising the primer according to item 20 above.
22. A kit for evaluating disease vulnerability, comprising the primer according to item 20 above.
23. A gene polymorphism marker for evaluating a tendency of an individual to have drug sensitivity, including a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism.
24. A gene polymorphism marker for evaluating the tendency of an individual to have disease vulnerability, comprising a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism.

  According to the present invention, an adrenergic receptor gene polymorphism capable of evaluating individual differences regarding drug sensitivity or disease vulnerability, or a haplotype constituted by the gene polymorphism, and drug sensitivity or disease using the gene polymorphism or haplotype Vulnerability evaluation methods can be provided. This evaluation method makes it possible to easily know or predict an appropriate prescription amount and an appropriate prescription schedule for narcotic drugs such as morphine, and is extremely useful for tailor-made pain treatment and drug dependence treatment.

Schematic showing gene polymorphisms identified for ADRB2 subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. Further, in the quadrangle in which the quadrangle that extends in the lower left or lower right direction from each SNP in the figure, the percentage of the calculated value of D ′, which is an index of linkage disequilibrium between SNP and SNP, is described. For example, the calculated value of D ′ between rs30312 and rs246503 is 0.98. Schematic showing gene polymorphisms identified for ADRB2 subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. In the rectangle intersect the rectangle leading to the lower left or lower right direction from each SNP in FIG, wrote the percentage of calculated values of r ^2, which is an index of linkage disequilibrium between SNP and SNP. For example, the calculated value of r ² between rs30312 and rs246503 is 0.80. Schematic showing gene polymorphisms identified for ADRB1 subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. Further, in the quadrangle in which the quadrangle that extends in the lower left or lower right direction from each SNP in the figure, the percentage of the calculated value of D ′, which is an index of linkage disequilibrium between SNP and SNP, is described. For example, the calculated value of D ′ between rs180914 and rs6585256 is 0.81. Schematic showing gene polymorphisms identified for ADRB1 subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. In the rectangle intersect the rectangle leading to the lower left or lower right direction from each SNP in FIG, wrote the percentage of calculated values of r ^2, which is an index of linkage disequilibrium between SNP and SNP. For example, the calculated value of r ² between rs180914 and rs6585256 is 0.56. The graph which shows the effect (the whole average +/- standard error) of ADRB2 subtype gene polymorphism (rs11959113) with respect to the administration required dose (microgram / kg) for 24 hours after an operation in the whole patient who received the analgesic in surgery. The graph which shows the effect (the whole average +/- standard error) of ADRB2 subtype gene polymorphism (rs2116719) with respect to the measurement result (second) of the preoperative pain sensitivity in the whole patient who received the analgesic in surgery. The graph which shows the effect (the whole average +/- standard error) of ADRB1 subtype gene polymorphism (rs1801252) with respect to the measurement result (second) of the preoperative pain sensitivity in the whole patient who received the analgesic in surgery. The graph which shows the effect (female average +/- standard error) of ADRB1 subtype gene polymorphism (rs1801253) with respect to the measurement result (second) of the pain sensitivity threshold value preoperatively in the female patient who received the analgesic in surgery. The graph which shows the effect (the male average +/- standard error) of ADRB1 subtype gene polymorphism (rs6585258) with respect to the measurement result (second) of the preoperative pain sensitivity in the male patient who received the analgesic in surgery. Effect of ADRB1 subtype polymorphism (rs6585258) on the measurement results (mm) of VAS (pain intensity) 24 hours after surgery in male patients who received analgesics in surgery (mean ± standard for men) Error). Schematic showing gene polymorphisms identified for ADRA2A subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. Further, in the quadrangle in which the quadrangle that extends in the lower left or lower right direction from each SNP in the figure, the percentage of the calculated value of D ′, which is an index of linkage disequilibrium between SNP and SNP, is described. For example, the calculated value of D ′ between rs990429 and rs491589 is 0.93. Schematic showing gene polymorphisms identified for ADRA2A subtype genes and linkage disequilibrium between them. In the figure, dark squares indicate strong SNPs. In the rectangle intersect the rectangle leading to the lower left or lower right direction from each SNP in FIG, wrote the percentage of calculated values of r ^2, which is an index of linkage disequilibrium between SNP and SNP. For example, the calculated value of r ² between rs990429 and rs491589 is 0.86. The effect of ADRA2A subtype gene polymorphism (rs7088234) on the total amount of necessary analgesic treatment (μg / kg) during and 24 hours after surgery in patients who received analgesics during surgery (overall mean ± standard error) ).

  Hereinafter, the present invention will be described in detail. However, the scope of the present invention is not limited to these descriptions, and modifications other than the following exemplifications can be made as appropriate without departing from the spirit of the present invention. This specification includes the entire specification of Japanese Patent Application No. 2010-198319, which is the basis for claiming priority of the present application. In addition, all publications cited in the present specification, for example, prior art documents, and publications, patent publications and other patent documents are incorporated herein by reference.

1. Summary of the Present Invention (1) Adrenergic Receptor Adrenergic receptor (ADR) is a G protein-coupled receptor activated by catecholamines such as adrenaline and noradrenaline. In addition to myocardium, various smooth muscles, fat cells, skeletal muscles, etc., it also exists in the brain. In addition to cardiac contraction, relaxation of various smooth muscles, basal metabolism, vasoconstriction, etc., analgesia and pain due to opioids etc. (Perception / Emotion) Side).

  Opioid receptors are present in brain stem periaqueductal gray, large raphe nuclei, giant cell reticular nucleus, para giant cell reticular nucleus, spinal dorsal horn, peripheral nerves, etc. The opioid action of gray matter is an analgesic effect by activating the descending pain inhibitory system leading to the dorsal horn of the spinal cord through serotonergic fibers from the major raphe nucleus and noradrenergic fibers from the para giant cell reticulated nucleus Demonstrate. Expression of adrenergic receptors has been observed in noradrenergic fibers.

  Here, the adrenergic receptor will be described. Adrenergic receptors are abundant in the brain and heart and play an important role in the control of neuronal excitability and heart rate. The adrenergic receptor has a structure that penetrates the cell membrane seven times. The receptor is coupled to the G protein.

Adrenergic receptors function in a dimeric state by combining two subunits. The types of receptor subtypes are roughly classified into α (α ₁ , α ₂ ), β (β ₁ , β ₂ , β ₃ ), and their subtypes are also known. The α subtype is strongly expressed in the brain, and the β subtype is also expressed. In the heart, the β subtype is strongly expressed.

(2) Gene polymorphism The present inventor, for healthy subjects, is a β subtype among α (α ₁ , α ₂ ), β (β ₁ , β ₂ , β ₃ ) subtypes that can constitute an adrenergic receptor. Genetic polymorphisms (SNP, etc.) were identified for (β ₁ , β ₂ ) (FIGS. 1 to 4). In addition, linkage disequilibrium analysis was performed as necessary to identify blocks with strong linkage disequilibrium (haplotype blocks).

  Here, linkage equilibrium refers to the case where the chromosomal relationship between two gene polymorphisms is independent, and linkage disequilibrium refers to Mendel's independent law because the gene polymorphism and gene polymorphism are linked. This is the case when deviating from the equilibrium state. In addition, the haplotype means a genetic configuration such as a gene or gene polymorphism adjacent to each other in one of a set of alleles (a gene derived from one parent).

  Genetic polymorphisms that are close together on the genome are inherited by haplotype blocks. In other words, it can be said that the haplotype is a combination of the arrangement of the same genes in the haplotype block.

  In the adrenergic receptor gene, when several gene polymorphisms appear in association with a phenotype, some gene polymorphisms constituting the haplotype can be changed without typing all individual gene polymorphisms. By analyzing, the relationship between the patient's genotype and phenotype can be clarified.

As a result of analysis of the adrenergic receptor β ₂ subtype gene, the present inventor found that 23 and 37 gene polymorphisms (see Table 3) were found in the 5 ′ and 3 ′ flanking regions, the translation region of exon 1 and Three and one (four total) gene polymorphisms (rs1801704, rs1042713, rs1042717 and rs1042718) were found in the 5 ′ untranslated region (see FIGS. 1 and 2).

For the β ₁ subtype gene, 21 and 8 gene polymorphisms (see Table 3) in the 5 ′ and 3 ′ flanking regions and two gene polymorphisms (rs1801252, rs1801253) in the translation region of exon 1 (See FIGS. 3 and 4).

  According to the present invention, by analyzing a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism, the number of analgesic administration necessary times, total analgesic drug amount, drug-dependent vulnerability, and stimulant It is possible to easily evaluate individual differences in drug vulnerability such as prolonged psychosis or disease vulnerability including pain sensitivity. The results of evaluating drug sensitivity or disease vulnerability are important information in determining the number of doses, dose and type of drug to be administered to an individual, and in predicting side effects. Therefore, the present invention is based on the analysis result of the gene polymorphism of the adrenergic receptor gene or the haplotype constituted by the gene polymorphism, and whether the individual (individual) has drug sensitivity or disease vulnerability (specifically, genetic The present invention provides a method for evaluating (specifically, knowing or predicting in advance) the tendency of whether or not there is a physical factor. The present invention also relates to the presence or absence of drug susceptibility or disease vulnerability of an individual (individual) including a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism (specifically, the presence or absence of a genetic factor) It is intended to provide a genetic polymorphism marker for evaluating the tendency (specifically, knowing or predicting in advance).

  In particular, since morphine and stimulants cause serious social problems depending on the method of use, it is important to be able to know or predict an appropriate dosage in advance for each individual before administration. Therefore, it can be said that the present invention is extremely useful in tailor-made pain treatment and drug-dependent treatment.

2. Adrenergic receptor gene polymorphism The human adrenergic receptor gene polymorphism of the present invention mainly includes single nucleotide polymorphism (hereinafter also referred to as SNP), but is not limited, insertion polymorphism, It may also include deletion polymorphisms and base repeat polymorphisms.

  A single nucleotide polymorphism [SNP (SNPs)] means a gene polymorphism resulting from substitution of one specific base of a gene with another base. The insertion / deletion polymorphism means a gene polymorphism caused by deletion / insertion of one or more bases.

  Further, the base repeat polymorphism means a gene polymorphism caused by the difference in the number of base sequence repeats. Base repeat polymorphism is classified into microsatellite polymorphism (number of bases: about 2 to 4 bases) and VNTR (variable number of tandem repeat) polymorphism (repetitive bases: several bases to several tens of bases) due to the difference in the number of repeated bases. The number of repetitions varies depending on the individual.

  Information on human adrenergic receptor gene polymorphisms revealed by the present invention (ADNP2 subtype gene, ADRB1 subtype gene and SNP in ADRA2A subtype gene found on the genome of Japanese healthy individuals) is shown in Table 3A and Shown in Table 3B. The gene polymorphisms shown in Table 3A and Table 3B are included in the adrenergic receptor gene polymorphisms of the present invention.

  In Table 3A and Table 3B, “ADRB2” (italic notation) represents the ADRB2 subtype gene (ADRB2 gene, which is a subtype of the adrenergic receptor gene), and “ADRB1” (italic notation) represents the ADRB1 subtype gene ( "ADRA2A" which is a subtype of the adrenergic receptor gene), and "ADRA2A" (italic notation) represents the ADRA2A subtype gene (the ADRA2A gene which is a subtype of the adrenergic receptor gene).

  “Position” means a position on the genome of an adrenergic receptor gene, and represents a 5 ′ flanking region, a 3 ′ flanking region, and an exon.

  The “gene polymorphism name” is the name of the SNP at the position on the genome and is registered in the dbSNP database (accessible from http://www.ncbi.nlm.nih.gov/projects/SNP/). Yes (same in this specification). Basically, the ID of “rs” is given before 4 or more digits to identify which SNP.

  The “major allele” and “minor allele” represent the major and minor alleles on the genome of Japanese healthy individuals, respectively.

The method for obtaining genetic polymorphism information in the present invention is, for example, as follows.
(1) Genomic DNA is purified from a blood sample collected from a human using the phenol method or the like. At that time, a commercially available genomic DNA extraction kit or apparatus such as GFX Genomic Blood DNA Purification Kit (manufactured by GE Healthcare Biosciences) may be used.
(2) Next, using the obtained genomic DNA as a template, the genomic DNA is divided into several parts by PCR, and used as a template DNA for sequencing. Since the present invention targets gene polymorphism, it is desirable to use an enzyme used in the PCR method with as high a fidelity as possible.
(3) The region around each gene polymorphism of the adrenergic receptor gene is amplified by PCR by about 500 to 1000 bp by using primers designed based on sequence information published in GenBank.
(4) The base sequence of the entire region of these PCR fragments is decoded by a sequencing method using a primer designed based on the sequence information published in GenBank, and the target gene polymorphism information is obtained. Obtainable.

Another method for obtaining genetic polymorphism information in the present invention is as follows.
(1) Genomic DNA is purified from a blood sample collected from a human using the phenol method or the like. At that time, a commercially available genomic DNA extraction kit or apparatus such as GFX Genomic Blood DNA Purification Kit (manufactured by GE Healthcare Biosciences) may be used.
(2) Next, the obtained genomic DNA is dissolved in TE buffer (10 mM tris-HCl, 1 mM EDTA, pH 8.0) and adjusted to 100 ng / μl.
(3) Genome-wide genotyping is performed according to the manufacturer's protocol by the Infinium assay II method using the iScan system (Illumina, San Diego, CA) manufactured by Illumina.
(4) Whole genome genotyping data analysis is performed using BeadStudio Genotyping module v3.3.7 (Illumina), etc., and quality control (Quality control) of gene polymorphism data of each sample is performed.
(5) From these whole genome genotyping data, select gene polymorphisms included in the gene region and flanking region using the annotation information of the target gene name as a clue, BeadStudio Genotyping module v3.3.7 (Illumina), etc. All the polymorphism information is extracted using the output function of.

  The present invention relates to ADRB2 subtype gene polymorphisms (rs888956, rs10075525, rs2116719, rs10059242, rs11742884, rs2116713, rs2400707, rs1042713, rs1042717, which can specifically hybridize to a DNA fragment containing a gene polymorphism of an adrenergic receptor gene. rs4705271, rs3857420, rs6888011, rs11959113, rs17707884, rs11742519, rs10214302, rs11168071, rs17640705, rs1432631, rs919725, rs1864932, rs11740830, rs4705283 and rs1181135), ADRB1 subtype polymorphisms (rs1801252, rs11 407, rs11 rs623499, rs2782979, rs2782980, rs12771397, rs11196592, rs11196597, rs10787516 and rs4918887) and ADRA2A subtype polymorphisms (rs990428, rs11195417, rs7074629, rs491589, rs34665740, rs745557, rs7905613, rs491863, rs7908645, rs7081 , Rs1537770, rs2032023, rs17128431, rs7069564, rs954385, rs1889745, rs1335706, rs1335704, rs1 335703, rs11195442, rs12220858, rs4545476, rs7908674, rs10885102, rs7084236, rs7088234 and rs7079973) are provided. The gene polymorphic site is the 51st base of the base sequence shown in any one of SEQ ID NOs: 1-38 and 47-76.

  The base of the oligonucleotide of the present invention preferably has at least 10 bases, preferably 10 to 150 bases, more preferably 10 to 45 bases, and still more preferably 14 to 25 bases.

  The oligonucleotide of the present invention includes, for example, the nucleotide sequence shown in any one of SEQ ID NOs: 1 to 38 and 47 to 76 containing the above-described gene polymorphism of the adrenergic receptor gene or complementary to the nucleotide sequence. Examples include oligonucleotides selected from the base sequences (Table 4A, Table 4B).

  The oligonucleotide of the present invention comprises 5. In the detection of an adrenergic receptor gene polymorphism described later in (2), it can be used as an adrenergic receptor gene-specific probe and primer.

  In Table 4A and Table 4B (SEQ ID NOs: 1-38 and 47-76), 101 bases are displayed, and the gene polymorphic site is displayed at the 51st position. For example, “A / G” means that the gene polymorphism is “A” and “G”, and “C / T” is the gene polymorphism of “C” and “T”. Means.

3. Haplotype analysis In the present invention, a haplotype can be constructed using SNP among the above-mentioned gene polymorphisms. The SNP to be subjected to haplotype analysis only needs to have a gene polymorphism frequency of 0.5% or more, preferably 1%, more preferably 5% or more. Further, the SNPs to be subjected to haplotype analysis may be all or a part thereof.

  Haplotype analysis can be analyzed by various computer programs, for example, available from the website of Haploview (http://www.broadinstitute.org/haploview/haploview (the same applies hereinafter); Barrett JC, Fry B , Maller J, Daly MJ. Haploview: analysis and visualization of LD and haplotype maps. Bioinformatics. 2005 Jan 15 [PubMed ID: 15297300] Whitehead Institute for Biomedical Research Cambridge, MA 02142, USA.).

  As an example of haplotype analysis, 2. Among the adrenergic receptor gene polymorphisms found in healthy Japanese, 24 SNPs that are ADRB2 subtype polymorphism, 12 SNPs that are ADRB1 subtype polymorphism, and ADRA2A subtype gene polymorphism Haploview was used to estimate haplotypes for 30 SNPs. The estimated haplotypes are shown in Table 5, Table 6A, and Table 6B, respectively.

Furthermore, the haplotype frequency in the population can be calculated from the genotype information of each individual's adrenergic receptor (ADR) gene in the population, and linkage disequilibrium analysis can be performed based on the haplotype frequency. The D ′ value and the r ² value, which are values indicating the scale of linkage disequilibrium, can be calculated based on the following definitions.

Definition:
There are SNP A and SNP B, and the alleles are A, a, B, and b. The four haplotypes made by SNP A and SNP B are AB, Ab, aB, ab, and the frequency of each haplotype is P _AB , P _Ab , P _aB , P _ab ,
D = P _AB × P _ab -P _Ab × P _aB (when D> 0)
D´ = (P _AB × P _ab -P _Ab × P _aB ) / Minimum ((((P _AB + P _aB ) × (P _aB + P _ab )), ((P _AB + P _Ab ) × (P _Ab + P _ab ))) (when D <0)
D´ = (P _AB × P _ab -P _Ab × P _aB ) / Minimum ((((P _AB + P _aB ) × (P _AB + P _Ab )), ((P _aB + P _ab ) × (P _Ab + P _ab )))
r ² = (P _AB × P _ab -P _Ab × P _aB ) ² / [(P _AB + P _Ab ) (P _AB + P _aB ) (P _aB + P _ab ) (P _Ab + P _ab )]] , Minimum (((P _AB + P _aB ) × (P _aB + P _ab )), ((P _AB + P _Ab ) × (P _Ab + P _ab ))) is (P _AB + P _aB ) × ( It means that the smaller one of (P _aB + P _ab ) and (P _AB + P _Ab ) × (P _Ab + P _ab ) is taken. ]

  Furthermore, haplotype blocks can be estimated from the results of linkage disequilibrium analysis. The haplotype block can be estimated from the result of the haplotype analysis using, for example, Haploview.

  By examining a specific SNP in the estimated haplotype block, it is possible to know information on SNPs that are indirectly linked in the same block. In other words, it is not necessary to analyze all SNPs when examining gene polymorphisms of adrenergic receptor genes (specifically, ADRB2 subtype gene, ADRB1 subtype gene or ADRA2A subtype gene). You only have to type.

4). Correlation between Adrenergic Receptor Gene Polymorphism and Drug Sensitivity or Disease Vulnerability It is considered that the function and expression level of an adrenergic receptor may change when a gene polymorphism occurs in the adrenergic receptor gene. Thus, there may be a correlation between adrenergic receptor gene polymorphisms and various phenotypes associated with adrenergic receptors.

  Here, examples of the phenotype include a phenotype related to drug sensitivity (drug sensitivity) and a phenotype related to disease onset (disease vulnerability). Drug sensitivity includes drug effectiveness, drug side effects, drug effective duration, and the like. Disease vulnerability includes pain sensitivity, vulnerability to drug dependence, and the like.

Examples of the drug include, but are not limited to, opioid receptor function modifiers and adrenergic receptor function modifiers. For example, various drugs that directly or indirectly act on opioid receptors or adrenergic receptors. Can be mentioned. As various drugs that act directly or indirectly on opioid receptors, specifically, stimulants such as methamphetamine, dopamine receptor agonists, dopamine receptor antagonists, μ, κ, δ opioid receptor agonists, Examples thereof include μ, κ, and δ opioid receptor antagonists. Specific drugs that directly or indirectly act on adrenergic receptors include catecholamines such as noradrenaline, tricyclic and tetracyclic antidepressants, selective norepinephrine reuptake inhibitors (NRIs), norepinephrine. -Dopamine reuptake inhibitors (NDRIs), serotonin / norepinephrine reuptake inhibitors (SNRIs), serotonin / norepinephrine / dopamine reuptake inhibitors (SNDRIs), stimulants such as amphetamines, natural resources such as Dutch Vieu, α ₁ , α ₂ , Β ₁ , β ₂ , β ₃ adrenergic receptor agonist, α ₁ , α ₂ , β ₁ , β ₂ , β ₃ adrenergic receptor antagonist and the like.

  Examples of opioid receptor function modifiers include, for example, morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, narolphine, levalorphan, pentazocine, pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, Etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, flurbiprofen axetil, marcaine, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, laughter, F3 (1-chloro-1,2 , 2 trifluorocyclobutane), halothane, estradiol, dithiothreitol, thioridazine, pimozide, fluoxetine, paroxy Cetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodir, bupivacaine, telthiapine, clozapine, haloperidol, SCH23390 and cocaine, etc. Prophenaxetyl and marcaine are preferred, and morphine, fentanyl and pentazocine are more preferred.

  Examples of adrenergic receptor function modifiers include, Amitraz, anisodamine, apraclonidine, brimonidine, silazoline, detomidine, dexmedetomidine, epinephrine, ethylephrine, indanidine, lofexidine, medetomidine, mephentermine, metallaminol, methoxamine, midodrine, mibazerol, propanoline, norphenamine octopamine Rilmenidine, Lomifiji , Synephrine, talipexol, tizanidine, dibenamine, ergotamine, ergomethrin, phentolamine, torazoline, bunazosin, naphthopidyl, prazosin, terazosin, doxazosin, tamsulosin, silodosin, alfuzosin, trimazosin, phenoxybenzampine, mitripriline, , Yohimbine, efaloxan, idazoxan, lauorcine, atipamezole, dobutamine, isoproterenol, xamoterol, epinephrine, isoprenaline, salbutamol (albuterol), levosalbutamol (leverbuterol), terbutaline, metaproterenol, fenoterol, vitorterol, probuterol, probuterol Salmeterol, fu Lumoterol, bambuterol, clenbuterol, isoetarine, ritodrine, vitorterol mesylate, L-796568, amibegron, solabegron, albutamine, befnolol, bromoacetylalprenolol mentane, broxaterol, cimaterol, silazoline, denopamine, dopexamine, hexiprenaline, heptrenaline , Mabuterol, methoxyphenamine, nilidrin, oxyfedrine, prenalterol, ractopamine, reproterol, limiterol, tretoquinol, tulobuterol, zilpaterol, ginterol, alprenolol, bucindolol, sotalol, bopindolol, pindolol, timolol, dichloroisoprenalin, alp Renolol, carteolol, Indenolol, bunitrolol, penbutolol, propranolol, nadolol, nipradilol, chirisolol, acebutolol, ceriprolol, metoprolol, atenolol, bisoprolol, betaxolol, practolol, bevantolol, esmolol, nebivolol, butoxamine, ICI-118,551, SR 59230, predrol Norolol, Landiolol, Levobunolol, Metipranolol, Labetalol, Carvedilol, Arotinolol, Amosuralol, Amphetamine, Droxidopa, Tyramine, Ephedrine, Pseudoephedrine, Atomoxetine (tomoxitine), Mazindol, Reboxetine, Viloxidine, Prominepdate Canfamin, Encamine, lefetamine, methylphenidate, piperdolol, prolintan, pyrovalerone, difemetrex, desvenlafaxine, duloxetine, milnacipran, venlafaxine, buttriptyline, desipramine, dosrepin, imipramine, lofepramine, nortriptyline, protriptyline , Amoxapine, maprotiline, mianserin, cyclobenzaprine, mesocarb, nefadozone, nefopam, sibutramine, tapentadol, tramadol, ziprasidone, adhiperifolin, hyperforin, cocaine, desoxypipradorol, diphenylprolinol, tyleneoxypylobarerone , Cyclazine Dole, Esreboxetine, Manifaxin, Nisoxetine, Radafaxin, Tandamine, WYE -103231, 1- (Indolin-1-yl) -1-phenyl-3-propan-2-olamines, 1- or 3- (3-Amino-2-hydroxy-1-phenyl propyl) -1,3-dihydro -2H-benzimidazol-2-ones, (+)-S-21, thienyl-based, bicifazine, brasofensin, diclofensin, DOV-21,947, DOV-102,677, DOV-216,303, indatraline, NS-2359, oxa Protilin, SEP-225,289, SEP-227,162, tesofensin and Dutchville are more preferred.

The correlation between the adrenergic receptor gene polymorphism and the phenotype can be examined, for example, as in the following (1) to (4).
(1) As a result of linkage disequilibrium analysis and haplotype analysis in healthy subjects, gene polymorphisms within the estimated linkage disequilibrium block are selected. For example, Tag SNP, which is a representative gene polymorphism, is selected as an adrenergic receptor gene polymorphism for correlation analysis with a phenotype.
(2) Next, the gene polymorphism frequency of the gene polymorphism in the subject (patient) is analyzed. When investigating the correlation between genetic polymorphism and disease vulnerability, comparison is made between the genetic polymorphism frequencies of the subject and the healthy subject. It is effective to use a statistical method such as chi ² square test in comparison.
Here, the subjects may be further classified according to phenotype differences, and the gene polymorphism frequencies and genotypes of healthy subjects and subjects may be compared for each classification. When the phenotype related to the onset of the disease is a stimulant psychosis-like symptom, for example, the period from the start of the use of a stimulant to the onset of delusions or hallucinations, the duration of delusions or hallucinations after cessation of use, presence or absence of relapse, multidrug abuse Can be classified by the presence or absence of.
(3) If there is a gene polymorphism significantly associated with drug sensitivity in a subject, the gene polymorphism can be used for evaluation of a genetic predisposition for drug sensitivity. In addition, if there is a genetic polymorphism having a significant difference in gene polymorphism frequency between a healthy person and a subject, the genetic polymorphism can be used for evaluation of a genetic predisposition for disease vulnerability.

  However, since it is suggested that the tendency of genetic polymorphism is influenced by race, birthplace, etc., genetic polymorphism similar to the population used to find related genetic polymorphism (eg SNP) It is desirable to perform the above evaluation using the gene polymorphism in a group showing

Specific examples of the correlation between the adrenergic receptor gene polymorphism and the phenotype are shown in (1) to (8) below.
(1) Patients who have undergone surgery with a major allele (G) in the ADRB2 subtype polymorphism (rs11959113) in the correlation with the measurement results of the 24 hour post-operative analgesic drug administration In correlation, the postoperative analgesic dose requirements were statistically significantly higher. Therefore, by analyzing the ADRB2 subtype gene polymorphism (rs11959113), the sensitivity to analgesics can be predicted more easily.

(2) Prevalence of minor allele (G) in the ADRB2 subtype polymorphism (rs2116719) and pain detection latency in the correlation with the measurement results of pain detection latency by immersion in ice water before hand surgery The relationship was shown. Therefore, the sensitivity to pain can be more easily predicted by analyzing the ADRB2 subtype gene polymorphism (rs2116719).

(3) Patients who have undergone surgery without major allele (A) with ADRB1 subtype polymorphism (rs1801252) in the correlation with measurement results of pain perception latency due to pre-operative immersion in ice water Compared with the patient group with allele A, the pain-sensing latency in immersion in hand ice water was statistically significantly longer. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs1801252), sensitivity to pain can be predicted more easily.

(4) Surgery without minor allele (G) in ADRB1 subtype polymorphism (rs1801253) in correlation with measurement results of pain detection latency threshold difference by immersion in hand ice water before and after fentanyl administration before surgery Compared with the female patient group with the allele G, the female patient group had a statistically significant difference in pain-sensing latency threshold value in hand ice water immersion. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs1801253), it is possible to more easily predict the sensitivity to analgesics.

(5) A group of male patients who underwent surgery without a minor allele (T) in the ADRB1 subtype gene polymorphism (rs6585258) in the correlation with the measurement results of pain perception latency due to pre-operative immersion in hand ice water Compared with the male patient group with the allele T, the pain-sensing latency threshold in the ice water immersion of the hands was statistically significantly shorter. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs6585258), sensitivity to pain can be predicted more easily.

(6) Surgery that does not have minor allele (T) in ADRB1 subtype gene polymorphism (rs6585258) in correlation with measurement results of pain intensity scale (VAS: Visual analogue scale) 24 hours after surgery The group of male patients who received the VAS value was statistically significantly greater than the group of male patients with the allele T. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs6585258), it is possible to more easily predict postoperative pain or sensitivity to analgesics.

(7) Patients who have undergone surgery with minor allele (G) in ADRA2A subtype polymorphism (rs7088234) in correlation with the measurement results of the total amount of analgesics administered 24 hours after surgery There was a statistically significant increase in the required amount of total analgesic during and after surgery. Therefore, by analyzing the ADRA2A subtype gene polymorphism (rs7088234), the sensitivity to analgesics can be predicted more easily.

(8) Intraoperative and surgical procedures for several ADRA2A subtype polymorphisms (rs11195417, rs7905613, rs7079291, rs7069564, rs4545476, rs10885102, rs7084236, rs7088234, rs7088234, rs7079973, rs7079973 and rs6585041), including other gene polymorphisms A statistically significant association was found with any of the following phenotypes: 24 hour total analgesic dose requirement, preoperative pain sensitivity, 24 hour postoperative pain sensitivity (VAS), and fentanyl analgesic effect. Therefore, by analyzing these genetic polymorphisms (rs11195417, rs7905613, rs7079291, rs7069564, rs4545476, rs10885102, rs7084236, rs7088234, rs7088234, rs7079973, rs7079973 and rs6585041), analgesic sensitivity, pain sensitivity, and fentanyl The analgesic effect can be predicted more easily.

5. Use of analysis results Correlation between adrenoceptor gene polymorphism and phenotype analyzed as described in 4 above is a method for predicting opioid receptors and the sensitivity of various drugs and pain related to adrenergic receptors, opioid receptors and It can be used as an index for a method for selecting a treatment or prevention method for a disease related to an adrenergic receptor, a method for determining an appropriate dose of a therapeutic drug, a method for predicting a side effect, or the like.

  Moreover, it is possible to evaluate the drug sensitivity or disease vulnerability by the race difference using the gene polymorphism or method of the present invention. The subject is not particularly limited, and examples include Japanese and Westerners. In the present invention, it is preferable that the subject has the same genetic polymorphism tendency as Japanese or Japanese.

6). Detection of genetic polymorphism A genomic sample from a test subject can be extracted from blood, saliva, skin, etc., but is not limited to this as long as the genomic sample can be collected. Methods for extracting and purifying genomic DNA are well known. For example, genomic DNA is purified from specimens such as blood, saliva and skin collected from humans using the phenol method or the like. At that time, a commercially available genomic DNA extraction kit or apparatus such as GFX Genomic Blood DNA Purification Kit (manufactured by GE Healthcare Biosciences) may be used. When the SNP to be detected is in an exon, mRNA or total RNA may be extracted instead of genomic DNA.

  The above-described oligonucleotide of the present invention can be used as a probe and a primer for detecting an adrenergic receptor gene polymorphism in a genomic sample. Hereinafter, an example of a genetic polymorphism detection method is shown.

(1) Detection of genetic polymorphism by PCR
In order to amplify a test sample by PCR, it is preferable to use a high-fidelity DNA polymerase such as KOD Dash polymerase (manufactured by TOYOBO). The primer to be used is designed and synthesized so that the target SNP in the test sample can be amplified. It is preferable that the gene polymorphism or its complementary strand is included at any position between the upstream and downstream primers. After completion of the amplification reaction, the amplification product is detected, and the presence or absence of the gene polymorphism is determined by a sequencing method or the like.

  Suitable primers that can be used in the method of the present invention include, for example, primers shown in Table 7 below.

(2) Detection of gene polymorphism by nucleotide sequencing method The gene polymorphism of the present invention can also be detected by nucleotide sequencing method based on dideoxy method. A commercially available ABI series (Applied Biosystems (Life Technologies)) can be used as a sequencer used for base sequence determination.

(3) Detection of genetic polymorphism by DNA microarray
The DNA microarray has an oligonucleotide probe immobilized on a support, and includes a DNA chip, a Gene chip, a microchip, a bead array, and the like. First, a polynucleotide of a test sample is isolated, amplified by PCR, and labeled with a fluorescent reporter group. Subsequently, labeled DNA / mRNA and total RNA are incubated with the array.

  Next, the array is inserted into a scanner, and a hybridization pattern is detected. Hybridization data is collected as luminescence from fluorescent reporter groups attached to the probe array (ie, incorporated into the target sequence). A probe that perfectly matches the target sequence produces a stronger signal than one that has a portion that does not match the target sequence. Since the sequence and position of each probe on the array is known, complementarity can determine the sequence of the target polynucleotide reacted with the probe array.

(4) Detection of genetic polymorphism by TaqMan PCR
TaqMan PCR is a method using fluorescently labeled allele-specific oligonucleotides (also called TaqMan probes (and PCR using Taq DNA polymerase). Allele-specific oligonucleotides are oligonucleotides that contain gene polymorphic sites. An allele-specific oligonucleotide used in the TaqMan PCR method can be designed based on the gene polymorphism information.

(5) Detection of gene polymorphism by invader method The invader method is a method for detecting a gene polymorphism by hybridization of an allele-specific oligonucleotide and a template. Kits for performing the invader method are commercially available (for example, Nano Invader® Array (manufactured by BML)), and the gene polymorphism can be easily detected by this method.

7). Kit The present invention provides a kit for evaluating drug / pain sensitivity or disease vulnerability. The gene polymorphism detection kit of the present invention contains one or more components necessary for carrying out the present invention.

  For example, the kit of the present invention preferably contains a reaction component necessary for storing or supplying the enzyme and / or performing genetic polymorphism detection. Such components include, but are not limited to, for example, the oligonucleotides of the present invention, enzyme buffers, dNTPs, control reagents (eg, tissue samples, target oligonucleotides for positive and negative controls), for labeling and And / or a detection reagent, a solid support, instructions, and the like. Further, the kit of the present invention may be a partial kit containing only a part of necessary components, and in that case, the user can prepare other components.

  The kit of the present invention can also be provided as a microarray in which the oligonucleotide is immobilized on a support. The microarray is obtained by fixing the oligonucleotide of the present invention on a support, and includes a DNA chip, a Gene chip, a microchip, a bead array, and the like.

  The kit of the present invention preferably contains an adrenergic receptor gene polymorphism found in the present invention and an oligonucleotide that can specifically hybridize to a DNA fragment containing the gene polymorphism.

  When determining a genetic polymorphism using the kit of the present invention, for example, blood is collected before the drug is used in a patient or the like (for example, before surgery, at the time of cancer pain, etc.), and DNA containing an adrenergic receptor gene is isolated, The gene is reacted with the oligonucleotide in the kit to determine the genotype.

  Moreover, the kit of this invention may contain the primer which consists of a base sequence shown to sequence number 39-46 used for adrenoceptor gene polymorphism detection. Detection of an adrenergic receptor gene polymorphism using the primer is performed, for example, by PCR.

  From the determined genotype and gene polymorphism, an administration plan such as the type or dose of the drug can be prepared. As a result, a drug effect suitable for an individual can be obtained, which is useful for tailor-made medical care. For example, when morphine is used, an analgesic effect suitable for an individual can be obtained and side effects can be minimized.

Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited to these examples.

<SNP analysis and haplotype construction>
(SNP analysis)
Genomic DNA is extracted from the blood of humans (Japanese healthy individuals: 121 people) by conventional methods, and genes for two subtypes (ADRB2, ADRB1) expressed in the heart and brain among many subtypes of human adrenergic receptors A polymorphism was identified.

  For the ADRB2 subtype gene, the entire exon region, 5 ′ and 3 ′ flanking regions, and part of the intron were included in the analysis range. The ADRB2 subtype gene is composed of one exon, and 4 gene polymorphisms were identified in Japanese samples, 3 and 1 respectively in the translation region and 5 ′ untranslation region of exon 1. In addition, 23 and 37 gene polymorphisms were found in the 5 ′ and 3 ′ flanking regions (see FIGS. 1, 2 and Table 8). As a result of linkage disequilibrium analysis, three linkage disequilibrium blocks were found in the 5 ′ flanking region, one linkage disequilibrium block was found in the exon 1 region, and six linkage disequilibrium blocks were found in the 3 ′ flanking region. Tag SNPs representative of this linkage disequilibrium block are rs888956, rs10075525, rs2116719, rs10059242, rs11742884, rs2116713, rs2400707, rs1042713, rs1042717, rs4705271, rs3857420, rs6888011, rs11959113, rs17707884, rs11742519, rs10214rs, rs111705, rs10214rs rs919725, rs1864932, rs11740830, rs4705283 and rs1181135 were found to be suitable.

  Similarly, regarding the ADRB1 subtype gene, the entire exon region, the 5 ′ and 3 ′ flanking regions, and a part of the intron were included in the analysis range. The ADRB1 subtype gene is composed of one exon, and two gene polymorphisms were identified in Japanese samples, two in the translation region of exon 1, 21 in the 5 ′ and 3 ′ flanking regions (FIG. 3). FIG. 4 and Table 8). As a result of linkage disequilibrium analysis, three linkage disequilibrium blocks were found from the 5 ′ flanking region to the exon 1 region and the 3 ′ flanking region, and one linkage disequilibrium block was found in the 3 ′ flanking region. As Tag SNPs representing this linkage disequilibrium block, rs1801252, rs1801253, rs1411407, rs6585258, rs2480792, rs11196589, rs623499, rs2782979, rs2782980, rs12771397, rs11196592, rs11196597, rs10787516 and rs4918887 were found to be suitable.

  In Table 8, the polymorphism causing amino acid substitution, that is, the polymorphism in which the type of amino acid after translation changes depending on the allele of the gene polymorphism was only rs1042713 of the ADRB2 gene.

  In Table 8, “minor allele frequency” means the ratio of minor alleles. The number of healthy subjects who were subjects was 103 in total for rs1801252 and rs1801253 of the ADRB1 gene, and 121 in total for other polymorphisms.

(Haplotype construction)
As an example of haplotype analysis, among the adrenergic receptor gene polymorphisms of healthy Japanese individuals, 24 SNPs that are ADRB2 subtype polymorphisms and 12 SNPs that are ADRB1 subtype polymorphisms shown in Table 8 , Haplotypes were estimated using Haploview. Table 9 and Table 10 show the estimated haplotypes.

  As shown in Table 9, at least 16 haplotypes were estimated as ADRB2 subtype polymorphism haplotypes in healthy Japanese individuals, of which 3 haplotypes were observed at a high frequency of 3% or more (haplotype numbers H1˜ H3). In addition, haplotypes estimated to occur with a frequency of less than 1% are collectively shown in Table 9 as haplotype numbers H17.

  Moreover, as shown in Table 10, at least 16 haplotypes were estimated as ADRB1 subtype polymorphism haplotypes in healthy Japanese individuals, of which 7 haplotypes were observed at a high frequency of 3% or more (haplotype number). H1-H7). Note that haplotypes estimated to occur at a frequency of less than 1% are collectively shown in Table 10 as haplotype numbers H17.

  In addition to the analysis of haplotype frequencies in the haplotype analysis shown in Table 9 and Table 10, linkage disequilibrium analysis was performed. The results are shown in FIGS. 1 and 2 show linkage disequilibrium between ADRB2 subtype gene polymorphisms in healthy Japanese subjects. 3 and 4 show linkage disequilibrium between ADRB1 subtype gene polymorphisms in Japanese healthy individuals.

  From the results of linkage disequilibrium analysis (FIGS. 1 to 4), linkage disequilibrium blocks were estimated using Haploview.

In Fig. 1, the D ′ value, which is an index of linkage disequilibrium between SNPs and SNPs, is calculated, and the value of 2 digits after the decimal point of the value is a quadrangle of squares that are connected to each SNP in the lower left or lower right direction. It is written in. Also, the r ² value, which is a stricter index of linkage disequilibrium, is calculated in the same manner, and the value is shown in the same square in FIG. A square having no value indicates that the value of D ′ or r ² is 1. 3 and 4 are similarly represented.

Focusing on D ′ in FIG. 1, complete linkage disequilibrium (D ′ = 1) was confirmed in many combinations of gene polymorphisms in the 5 ′ flanking region and the 3 ′ flanking region. Further, it was found that even in view of the r ² value of FIG. 2, a number of genetic polymorphisms strong linkage disequilibrium (r ² = 1). Tag SNPs representative of these linkage disequilibrium blocks are rs888956, rs10075525, rs2116719, rs10059242, rs11742884, rs2116713, rs2400707, rs1042713, rs1042717, rs4705271, rs3857420, rs6888011, rs11959113, rs17707884, rs11742519, rs10214rs, 143705, , Rs919725, rs1864932, rs11740830, rs4705283 and rs1181135 have been found to be suitable.

Further, focusing on D ′ in FIG. 3, complete linkage disequilibrium (D ′ = 1) was confirmed in many combinations of gene polymorphisms. Further, focusing on the r ² value in FIG. 4, it was found that several gene polymorphisms have strong linkage disequilibrium (r ² = 1). As tag SNPs representing these linkage disequilibrium blocks, rs1411407, rs6585258, rs2480792, rs11196589, rs623499, rs2782979, rs2782980, rs12771397, rs11196592, rs11196597, rs10787516 and rs4918887 were found to be suitable.

Moreover, the linkage disequilibrium block was estimated from the result (FIGS. 1-4) of the linkage disequilibrium analysis using Haploview. As a result, for the SNP of the ADRB2 subtype gene shown in FIG. 1 and FIG. 2, three linkage disequilibrium blocks in the 5 ′ flanking region, one linkage disequilibrium block in the exon 1 region, and the 3 ′ flanking region 6 linkage disequilibrium blocks were identified. Similarly, in the ADRB1 subtype gene SNP shown in FIGS. 3 and 4, three linkage disequilibrium blocks from the 5 ′ flanking region to the exon 1 region and the 3 ′ flanking region are represented by 1 in the 3 ′ flanking region. Two linkage disequilibrium blocks were identified.

<Correlation between ADRB2 subtype polymorphism (rs11959113) and analgesic administration requirement>
The correlation between the adrenergic receptor gene polymorphism and the required amount of analgesics was examined. Genomic DNA was extracted from the blood of 234 patients who underwent surgery, and one gene polymorphism (rs11959113) of ADRB2 subtype gene was determined. And the correlation with the determination result of these gene polymorphisms and the postoperative analgesic administration required amount was analyzed.

  As the analgesic, fentanyl, which was intravenously administered mainly by a PCA (Patient-controlled analgesia) pump, was used.

  As a result, as shown in Table 11 and FIG. 5 below, patients undergoing surgery with a major allele (G) in the ADRB2 subtype gene polymorphism (rs11959113) correlate with the number of alleles, and postoperatively The required dose of analgesics was statistically significantly higher. Therefore, by analyzing the ADRB2 subtype gene polymorphism (rs11959113), sensitivity to analgesics can be predicted.

  Based on the median value of 2.893 (μg) of fentanyl administration required 24 hours after surgery, the patient groups with smaller and larger values are defined as analgesic hypersensitive group and analgesic hyposensitive group, respectively. When stratified by the rs11959113 polymorphism in the ADRB2 gene, 22% and 78% of patients with A / A in this polymorphism were determined to be the analgesic hypersensitive group and the analgesic hyposensitive group, respectively. In contrast, in the G / G patient group, 56% and 44% of patients were classified as the analgesic hypersensitive group and the analgesic hyposensitive group, respectively.

<Correlation between ADRB2 subtype polymorphism (rs2116719) and pain sensitivity>
The correlation between adrenergic receptor gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 234 patients undergoing surgery, and one gene polymorphism (rs2116719) of the ADRB2 subtype gene was determined. And the correlation with the determination result of these gene polymorphisms and the measurement of the pain detection latency by ice water immersion before the operation was analyzed.

  As a result, as shown in Table 12 and FIG. 6 below, patients who have undergone surgery with a major allele (A) with ADRB2 subtype polymorphism (rs2116719) correlate with the number of alleles, and perceive pain. Latency measurement results (logarithmic transformation) were statistically significantly smaller. Therefore, the sensitivity to pain can be predicted by analyzing the ADRB2 subtype gene polymorphism (rs2116719).

  Based on the median value of 14 (seconds) of the measurement result of pain perception by immersion in ice water before surgery, the patient groups with lower and higher values were classified as pain high sensitivity group and pain low sensitivity group, respectively. Defined and stratified by the rs2116719 polymorphism of the ADRB2 gene, 58% and 42% of patients with A / A in this polymorphism were identified as the hypersensitive group and the hyposensitive group, respectively. On the other hand, in the G / G patient group, 25% and 75% of the patients were classified as a pain-sensitive group and a pain-insensitive group, respectively.

<Correlation between ADRB1 subtype gene polymorphism (rs1801252) and pain sensitivity>
The correlation between adrenergic receptor gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 216 patients undergoing surgery, and one gene polymorphism (rs1801252) of the ADRB1 subtype gene was determined. And the correlation with the determination result of these gene polymorphisms and the measurement of the pain detection latency by ice water immersion before the operation was analyzed.

  As a result, as shown in Table 13 and FIG. 7 below, the ADRB1 subtype gene polymorphism (rs1801252) has a major allele (A) in the correlation with the measurement result of pain perception latency by immersion in pre-operative ice water. The group of patients who had no surgery had a statistically significantly longer pain-sensing latency in the ice water immersion of hands compared to the group of patients with the allele A. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs1801252), sensitivity to pain can be predicted more easily.

  In addition, regarding the measurement result of pain perception latency by pre-operative hand ice water immersion, stratified by rs1801252 polymorphism of ADRB1 gene, 49% in the A / A or A / G patient group in this polymorphism and 51% of patients were determined to be pain sensitive groups and low pain sensitive groups, respectively, whereas 0% and 100% of patients in the G / G group were pain sensitive and pain insensitive groups, respectively. It was determined.

<Correlation between ADRB1 subtype polymorphism (rs1801253) and analgesic sensitivity>
We investigated the correlation between adrenergic receptor gene polymorphism and analgesic sensitivity. Genomic DNA was extracted from the blood of 216 patients undergoing surgery, and one gene polymorphism (rs1801253) of the ADRB1 subtype gene was determined. And the correlation with the determination result of these gene polymorphisms and the measurement of the pain detection latency by ice water immersion before the operation was analyzed.

  As a result, as shown in Table 14 and FIG. 8 below, the ADRB1 subtype gene polymorphism (rs1801253) is minor in the correlation with the measurement result of the difference in pain perception latency threshold due to immersion in hand ice water before and after fentanyl administration before surgery. The group of female patients who did not have an allele (G) had a statistically significantly longer pain-sensing latency threshold difference in ice water immersion than the group of female patients with the allele G. It was. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs1801253), it is possible to more easily predict the sensitivity to analgesics.

  In addition, in women, patients with smaller and larger values were measured based on the median value of 9 (seconds) of the results of measurement of the difference in pain perception latency by immersion in ice water before and after fentanyl administration before surgery. The analgesic hyposensitivity group and the analgesic hypersensitivity group were defined and stratified by the rs1801253 polymorphism of the ADRB1 gene. In this polymorphism, 39% and 61% of patients were analgesic in the C / C patient group, respectively. In the C / G or G / G patient group, 62.5% and 37.5% of patients were analgesic hyposensitive group and analgesic hypersensitive group, respectively. It was determined.

<Correlation between ADRB1 subtype gene polymorphism (rs6585258) and pain sensitivity>
The correlation between adrenergic receptor gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 234 patients who underwent surgery, and one gene polymorphism (rs6585258) of ADRB1 subtype gene was determined. And the correlation with the determination result of these gene polymorphisms and the measurement of the pain detection latency by ice water immersion before the operation was analyzed.

  As a result, as shown in Table 15 and FIG. 9 below, the ADRB1 subtype gene polymorphism (rs6585258) has a minor allele (T) in the correlation with the measurement result of pain perception latency by immersion in pre-operative hand ice water. The group of male patients who received no surgery had a statistically significantly lower pain-sensing latency threshold in hand ice water immersion than the male patient group with the allele T. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs6585258), sensitivity to pain can be predicted more easily.

  In addition, regarding the measurement results of pain perception latency by pre-operative ice water immersion in men, when stratified by the rs6585258 polymorphism of ADRB1 gene, 69% and 31% in the G / G patient group in this polymorphism % Of patients were determined to be pain sensitive and pain insensitive groups, whereas 35% and 65% of patients in the T / T or T / G patient groups were pain sensitive and pain low, respectively. It was judged as a sensitive group.

<Correlation between ADRB1 subtype gene polymorphism (rs6585258) and pain sensitivity>
The correlation between adrenergic receptor gene polymorphism and pain sensitivity was examined. Genomic DNA was extracted from the blood of 234 patients who underwent surgery, and one gene polymorphism (rs6585258) of ADRB1 subtype gene was determined. Then, the correlation between the determination result of these gene polymorphisms and the measurement of the pain intensity scale (VAS: Visual analogue scale) 24 hours after the operation was analyzed.

  As a result, as shown in the following Table 16 and FIG. 10, in the correlation with the measurement result of the pain intensity scale (VAS) 24 hours after the operation, the ADRB1 subtype gene polymorphism (rs6585258) is a minor allele (rs6585258). The group of male patients who received surgery without T) had a statistically significant VAS value compared to the male patient group with the allele T. Therefore, by analyzing the ADRB1 subtype gene polymorphism (rs6585258), it is possible to more easily predict postoperative pain or sensitivity to analgesics.

  In males, the patient groups with lower and higher pain values were defined as the low pain and high sensitivity groups, respectively, based on the VAS median value of 25 (mm) 24 hours after surgery. When stratified by the rs6585258 polymorphism of the gene, 30% and 70% of patients with G / G in the polymorphism were determined to be pain-insensitive groups and pain-sensitive groups, respectively. In the T / T or T / G patient group, 64% and 36% of patients were classified as the pain insensitive group and the pain sensitive group, respectively.

<SNP analysis and haplotype construction>
(SNP analysis)
Genomic DNA is extracted from the blood of humans (healthy Japanese: 127 people) in a conventional manner. Among many subtypes of human adrenergic receptors, two subtypes (ADRA2A) expressed in the heart and brain are polymorphic. The type was identified.

  For the ADRA2A subtype gene, the entire exon region, 5 ′ and 3 ′ flanking regions were included in the analysis range. The ADRA2A subtype gene consists of one exon, and four polymorphisms were identified in Japanese samples, one in the translation region of exon 1 and one in the 5 ′ and 3 ′ untranslated regions, respectively. . In addition, 13 and 46 gene polymorphisms were found in the 5 ′ and 3 ′ flanking regions (see FIGS. 11, 12 and Table 17). As a result of linkage disequilibrium analysis, one linkage disequilibrium block was found from the 5 ′ flanking region to the exon 1 region and the 3 ′ flanking region, and seven linkage disequilibrium blocks were found in the 3 ′ flanking region. Tag SNPs representing this linkage disequilibrium block include rs990428, rs11195417, rs7074629, rs491589, rs34665740, rs745557, rs7905613, rs4918621, rs7908645, rs7084501, rs1343450, rs1556716, rs7897445, rs1537770, rs17320431, rs7069564, rs495 rs1335706, rs1335704, rs1335703, rs11195442, rs12220858, rs4545476, rs7908674, rs10885102, rs7084236, rs7088234 and rs7079973 were found to be suitable.

In Table 17, there was no polymorphism causing amino acid substitution, that is, a polymorphism in which the type of amino acid after translation changed depending on the allele of the gene polymorphism.

In Table 17, “minor allele frequency” means the ratio of minor alleles.
The total number of healthy subjects who became subjects was 127.

(Haplotype construction)
As an example of haplotype analysis, among the adrenergic receptor gene polymorphisms of healthy Japanese individuals,
Haploview was used to estimate haplotypes for 63 SNPs that are ADRA2A subtype gene polymorphisms shown in Table 17. Table 18 shows the estimated haplotypes.

  As shown in Table 18, at least 27 haplotypes were estimated as ADRA2A subtype gene polymorphism haplotypes in healthy Japanese individuals, of which 7 haplotypes were observed at a high frequency of 3% or more (haplotype numbers H1 to H1). H7). Note that haplotypes estimated to occur at a frequency of less than 1% are collectively shown in haplotype number H28 in Table 18.

  A linkage disequilibrium analysis was performed along with the analysis of the haplotype frequency in the haplotype analysis shown in Table 18. As a result, linkage disequilibrium between ADRA2A subtype gene polymorphisms in healthy Japanese individuals is shown in FIG. 11 and FIG.

  From the results of linkage disequilibrium analysis (FIGS. 11 and 12), linkage disequilibrium blocks were estimated using Haploview.

In FIG. 11, a D ′ value, which is an index of linkage disequilibrium between SNPs and SNPs, is calculated, and the value of two digits after the decimal point of the value is a quadrangle formed by a quadrangle that extends from each SNP to the lower left or lower right direction. It is written in. Also, the r ² value, which is a stricter indicator of linkage disequilibrium, is calculated in the same manner, and the value is shown in the same square in FIG. A square having no value indicates that the value of D ′ or r ² is 1.

Focusing on D ′ in FIG. 11, complete linkage disequilibrium (D ′ = 1) was confirmed in many combinations of gene polymorphisms in the 5 ′ flanking region and the 3 ′ flanking region. Further, it was found that even in view of the r ² value of FIG. 12, a number of genetic polymorphisms strong linkage disequilibrium (r ² = 1). Tag SNPs representing these linkage disequilibrium blocks are rs990428, rs11195417rs7074629, rs491589, rs34665740, rs745557, rs7905613, rs4918621, rs7908645, rs7084501rs1343450, rs1556716, rs7897445, rs1537770, rs2032023, rs17128431, rs70695188, 704 , Rs11195442, rs12220858, rs4545476, rs7908674rs10885102, rs7084236, rs7088234 and rs7079973 were found to be suitable.

In addition, linkage disequilibrium blocks were estimated from the results of linkage disequilibrium analysis (FIGS. 11 and 12) using Haploview. As a result, for the SNP of the ADRA2A subtype gene shown in FIGS. 11 and 12, one linkage disequilibrium book from the 5 ′ flanking region to the exon 1 region and the 3 ′ flanking region was transferred to the 3 ′ flanking region. Two linkage disequilibrium blocks were identified.

<Correlation between ADRA2A subtype gene polymorphism (rs7088234) and total dose required for analgesics>
The correlation between the adrenergic receptor gene polymorphism and the total dose required for analgesics was examined. Genomic DNA was extracted from the blood of 247 patients undergoing surgery, and one gene polymorphism (rs7088234) of the ADRA2A subtype gene was determined. Then, the correlation between the determination results of these gene polymorphisms and the required amount of total analgesic during and after surgery was analyzed.

  As the analgesic, fentanyl, which was intravenously administered mainly by a PCA (Patient-controlled analgesia) pump, was used.

  As a result, as shown in Table 19 and FIG. 13 below, patients who have undergone surgery with ADRA2A subtype gene polymorphism (rs7088234) minor allele (G) are correlated with the number of alleles. The post-total analgesic required dose was statistically significantly higher. Therefore, by analyzing the ADRA2A subtype gene polymorphism (rs7088234), sensitivity to analgesics can be predicted.

  In addition, patients with higher and lower analgesic groups than those with a median value of 8.000 (μg) of the total amount of analgesic administration required during and after surgery for 24 hours. And stratified by the rs7088234 polymorphism of the ADRA2A gene.In this polymorphism, 54% and 46% of patients with A / A were classified as an analgesic hypersensitive group and an analgesic hyposensitive group, respectively. In contrast, in the G / G patient group, 32% and 68% of patients were classified as the analgesic hypersensitive group and the analgesic hyposensitive group, respectively.

<Correlation between ADRA2A subtype gene polymorphism and total analgesic dose requirement, pain sensitivity, and analgesic effect of fentanyl>
Similar to Example 9, adrenergic receptor gene polymorphism (rs11195417, rs7905613, rs7079291, rs7069564, rs4545476, rs10885102, rs7084236, rs7088234, rs70788234, rs7079973, rs7079973 and rs6585041) and total analgesia 24 hours after surgery The correlation between drug administration requirements, pain sensitivity, and the analgesic effect of fentanyl was investigated. Genomic DNA was extracted from the blood of a total of 361 patients who underwent surgery, and gene polymorphisms of the ADRA2A subtype gene (Tag SNP within the linkage disequilibrium block and individual SNP outside the linkage disequilibrium block) were determined. Analyzes the correlation between the results of these polymorphisms and the required amount of analgesic during and after surgery, preoperative pain sensitivity, postoperative pain sensitivity (VAS), and the analgesic effect of fentanyl did.

  As the analgesic, fentanyl, which was intravenously administered mainly by a PCA (Patient-controlled analgesia) pump, was used.

  As a result, as shown in Table 20 below (the results of Example 9 are also shown collectively), the ADRA2A subtype gene polymorphism (rs11195417, rs7905613, rs7079291, rs7069564, rs4545476, rs10885102, rs7084236, rs7088234, rs7088234, rs7079973, rs7079973 And rs6585041), the phenotype and statistical analysis of the total amount of analgesics required during and after surgery, preoperative pain sensitivity, pain sensitivity after 24 hours (VAS), and fentanyl analgesic effect A significant association was shown. Therefore, by analyzing these gene polymorphisms, sensitivity to analgesics, pain sensitivity, and analgesic effect of fentanyl can be predicted.

SEQ ID NO: 39: synthetic DNA
SEQ ID NO: 40: synthetic DNA
SEQ ID NO: 41: synthetic DNA
SEQ ID NO: 42: synthetic DNA
SEQ ID NO: 43: synthetic DNA
SEQ ID NO: 44: synthetic DNA
SEQ ID NO: 45: synthetic DNA
SEQ ID NO: 46: synthetic DNA

Claims (22)

  A method for evaluating drug sensitivity, comprising associating a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism with an individual drug sensitivity.   The method according to claim 1, wherein a tendency of the individual to have drug sensitivity is evaluated based on the analysis result of the gene polymorphism or the haplotype. The following steps:
(1) A step of performing linkage disequilibrium analysis and haplotype analysis in a healthy person, and selecting a gene polymorphism in the linkage disequilibrium block;
(2) analyzing the association between the genotype of the gene polymorphism in the subject and drug sensitivity, and (3) using the gene polymorphism significantly associated with drug sensitivity in the subject for evaluating drug sensitivity. The method according to claim 1 or 2, comprising.   A method for evaluating disease vulnerability, comprising associating a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism with a disease vulnerability of an individual.   5. The method according to claim 4, wherein a tendency of the presence or absence of disease vulnerability of an individual is evaluated based on the analysis result of the gene polymorphism or the haplotype. The following steps:
(1) A step of performing linkage disequilibrium analysis and haplotype analysis in a healthy person, and selecting a gene polymorphism in the linkage disequilibrium block;
(2) a step of comparing the gene polymorphism frequency of the gene polymorphism in the subject with the gene polymorphism frequency of the gene polymorphism in a healthy person, and (3) a significant difference in the gene polymorphism frequency between the subject and the healthy person The method of Claim 4 or 5 including the process of using a certain gene polymorphism for evaluation of disease vulnerability.   The method according to any one of claims 4 to 6, wherein the disease vulnerability is pain sensitivity or drug dependency.   The gene polymorphism is at least one selected from the group consisting of a single nucleotide polymorphism, an insertion polymorphism, a deletion polymorphism, and a nucleotide repeat polymorphism, according to any one of claims 1 to 7. the method of.   The gene polymorphism is the ADRB2 subtype genes rs888956, rs10075525, rs2116719, rs10059242, rs11742884, rs2116713, rs2400707, rs1042713, rs1042717, rs4705271, rs3857420, rs6888011, rs11959113, rs17707884, rs11742519, rs10214rs, 1711171, 919 rs1864932, rs11740830, rs4705283 and rs1181135, ADRB1 subtype genes rs1801252, rs1801253, rs1411407, rs6585258, rs2480792, rs11196589, rs623499, rs2782979, rs2782980, rs12771397, rs11196592, rs11196597, rs10787516, and rs4987587 , Rs11195417, rs7074629, rs491589, rs34665740, rs745557, rs7905613, rs4918621, rs7908645, rs7084501, rs1343450, rs1556716, rs7897445, rs1537770, rs2032023, rs17128431, rs7069564, rs954385, rs3354, rs3354, rs3354 , Rs10885102, rs7084236, rs7088234 and rs7079973 Kutomo is one method according to any one of claims 1-8. The method according to any one of claims 1 to 9, wherein the haplotype is at least one selected from those shown in the following table.
  A method for determining the type, amount, and / or number of administrations of a drug to be administered to an individual using the result evaluated by the method according to any one of claims 1 to 10 as an index.   The method of estimating the side effect of the drug administered to an individual | organism | solid using the result evaluated by the method of any one of Claims 1-10 as an parameter | index.   The method according to any one of claims 1 to 3, 7, 11 and 12, wherein the drug is an opioid receptor function modifier and / or an adrenergic receptor function modifier.   Opioid receptor function modifiers include methamphetamine, methylenedioxymethamphetamine, amphetamine, dextroamphetamine, dopamine, morphine, DAMGO, codeine, methadone, carfentanil, fentanyl, heroin, cocaine, naloxone, naltrexone, nalolphine, levalorphan Pethidine, buprenorphine, oxycodone, hydrocodone, levorphanol, etorphine, dihydroetorphine, hydromorphone, oxymorphone, tramadol, diclofenac, indomethacin, flurbiprofen axetil, marcaine, ethanol, methanol, diethyl ether, propanol, butanol, flupirtine, Lolita, F3 (1-chloro-1,2,2trifluorocyclobutane), haloce , Estradiol, dithiothreitol, thioridazine, pimozide, fluoxetine, paroxetine, desipramine, imipramine, clomipramine, tetramide, isoflurane, ginsenoside, ifenprodir, bupivacaine, telthiapine, clozapine, haloperidol, SCH23390 and cocaine Adrenergic receptor function modifier is adrenaline, noradrenaline, dopamine, phenylephrine, naphazoline, α-methylnoradrenaline, oxymetazoline, xylometazoline, clonidine, guanfacine, guanabenz, guanoxabenz, guanethidine, xylazine, Fadolmidine, amidephrine, amitraz, anisodamine, apraclonidine , Brimonidine, silazoline, detomidine, dexmedetomidine, epinephrine, ethylephrine, indanidine, lofexidine, medetomidine, mephentermine, metallaminol, methoxamine, midodrine, mibazelol, norphenephrine, octopamine, oxymetazoline, phenylpropanolamine, furimidine Talipexol, tizanidine, dibenamine, ergotamine, ergomethrin, phentolamine, torazoline, bunazosin, naphthopidyl, prazosin, terazosin, doxazosin, tamsulosin, silodosin, alfuzosin, trimazosin, phenoxybenzamine, amitriptyline, clomipramine, dopine Efaloxa , Idazoxan, lauorcin, atipamezole, dobutamine, isoproterenol, xamoterol, epinephrine, isoprenaline, salbutamol (albuterol), levosalbutamol (levolbuterol), terbutaline, metaproterenol, fenoterol, bitorterol, pyrbuterol, procaterol, salmeterol, , Bambuterol, clenbuterol, isoethalin, ritodrine, vitorterol mesylate, L-796568, amibegron, sorabegron, albutamine, befnolol, bromoacetylalprenololmenthane, broxaterol, simaterol, silazoline, denopamine, dopexamine, ethipremine, hexephrine Mabuterol, methoxy Enamine, nilidrin, oxyfedrine, prenalterol, ractopamine, reproterol, limiterol, tretoquinol, tulobuterol, zilpaterol, ginterol, alprenolol, bucindolol, sotalol, bopindolol, pindolol, timolol, dichloroisoprenalin, alprenolol, carteolol , Indenolol, bunitrolol, penbutolol, propranolol, nadolol, nipradilol, chirisolol, acebutolol, ceriprolol, metoprolol, atenolol, bisoprolol, betaxolol, practolol, bevantolol, esmolol, nebivolol, butoxamine, ICI-118,551, SR Sprenolol, Landiolol, Levobunoro , Metipranolol, labetalol, carvedilol, arotinolol, amosuralol, amphetamine, droxidopa, tyramine, ephedrine, pseudoephedrine, atomoxetine (tomoxitine), mazindol, reboxetine, viloxazine, amineptine, bupropionate, dexamphamifammine , Lepetamine, methylphenidate, piperdolol, prolintane, pyrovalerone, difemetrex, desvenlafaxine, duloxetine, milnacipran, venlafaxine, buttriptyline, desipramine, dosrepine, imipramine, lofepramine, nortriptyline, protriptyline, Amoxapine, maprotiline, mianserin, cyclobenzaprine, me Carb, nefadozone, nefopam, sibutramine, tapentadol, tramadol, ziprasidone, adhiperifolin, hyperforin, cocaine, desoxypipradorol, diphenylprolinol, tylenedioxypylobalerone, cyclazine, esreboxetine, manifaxin, nisoxetine, Radafaxine, tandamine, WYE-103231, 1- (Indolin-1-yl) -1-phenyl-3-propan-2-olamines, 1- or 3- (3-Amino-2-hydroxy-1-phenyl propyl)- 1,3-dihydro-2H-benzimidazol-2-ones, (+)-S-21, thienyl-based, bicifazine, brasofensin, diclofensin, DOV-21,947, DOV-102,677, DOV-216,303, indatraline, 14. The method according to claim 13, wherein the method is at least one selected from the group consisting of NS-2359, oxaprotiline, SEP-225,289, SEP-227,162, tesofensin and Dutchville.   At least the 51st base in the base sequence shown in any one of SEQ ID NOs: 1-38 and 47-76, which can specifically hybridize to a DNA fragment containing a gene polymorphism of an adrenergic receptor gene The method according to any one of claims 1 to 14, wherein an oligonucleotide comprising a base sequence having a length of 10 bases or a base sequence complementary to the base sequence is used.   The method according to claim 15, wherein the oligonucleotide has a length of 10 to 150 bases.   The oligonucleotide is an oligonucleotide selected from the group consisting of a base sequence represented by any one of SEQ ID NOs: 1-38 and 47-76 or a base sequence complementary to the base sequence. The method described in 1.   The primer which consists of a base sequence shown by any one of sequence number 39-46 used for the detection of the adrenergic receptor gene polymorphism in the method of any one of Claims 1-17.   A drug sensitivity evaluation kit comprising the primer according to claim 18.   A kit for evaluating disease vulnerability, comprising the primer according to claim 18.   A gene polymorphism marker for evaluating a tendency of an individual to have drug sensitivity, including a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism.   A gene polymorphism marker for evaluating the tendency of an individual to have disease vulnerability, comprising a gene polymorphism of an adrenergic receptor gene or a haplotype constituted by the gene polymorphism.