KR101591603B1 - Method of providing the information for predicting adverse drug reaction - Google Patents

Abstract

The present invention relates to a method for predicting an adverse drug reaction caused by anticonvulsants by detecting the presence of HLA-B*4403 in a biological specimen isolated from a patient. According to the present invention, the method for predicting the adverse drug reaction applies an HLA-B genotype detection method which has been widely and conventionally applied, leading to significant reduction in pain and risks due to the adverse drug reaction in the patient with reduced costs in a short amount of time. Thus, it is anticipated that the method of the present invention can be easily used to the patient prior to taking a medication.

Description

[0001] The present invention relates to a method for predicting adverse drug reactions,

The present invention relates to a method for predicting adverse drug reactions by anticonvulsants by confirming the presence of HLA-B * 4403 from a biological sample isolated from a patient.

The major histocompatibility complex (MHC) is an antigen system that is regulated in the gene domain. It refers to a group of genes involved in the rejection (immune response) that occurs during organ transplantation and exists in all mammals. Human primary histocompatibility complex was first found in an anti-leukocyte factor that recognizes an antigen on the surface of leukocytes. It is known as human leukocyte antigen (HLA) and is located at 6q21.3 of chromosome 6. These human leukocyte antigens have more than a dozen or more alleles and thus exhibit the highest genetic polymorphism among the many genetic markers in the human body, . Recently, studies have been actively conducted on the genotype of these human leukocyte antigens, which are known to be associated with adverse drug reactions (ADR) (Pharmaceuticals (2011) 4: 69-90).

Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rarely caused by serious adverse drug reactions caused by drug harmful reactions, but the mortality rate is 40% Is a very bad prognosis. Thus, if we can predict the risk of developing certain drugs associated with Stevens-Johnson syndrome and toxic epidermal necrolysis, we can significantly reduce the risk to patients and reduce the cost of medical expenses due to unwanted drug harm. However, there is no research on this in Korea yet. Moreover, genetic testing before drug administration is a non-payment item that is not covered by the national health insurance. to be.

Thus, in order to reduce the suffering and the risk of the patient due to severe adverse drug reactions, it is required to develop a method for predicting the risk of adverse reaction to a specific drug at a low cost in a short time.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems of the prior art, and it is an object of the present invention to provide a method of predicting drug- And the like.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, for purposes of complete understanding of the present invention, various specific details are set forth, such as specific forms, compositions, and processes, and the like. However, certain embodiments may be practiced without one or more of these specific details, or with other known methods and forms. In other instances, well-known processes and techniques of manufacture are not described in any detail, in order not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "embodiment" means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in at least one embodiment of the invention. Accordingly, the appearances of the phrase " in one embodiment "or" an embodiment "in various places throughout this specification are not necessarily indicative of the same embodiment of the present invention. In addition, a particular feature, form, composition, or characteristic may be combined in any suitable manner in one or more embodiments.

As used herein, the term "adverse drug reaction " refers to the broad concept of unintended response by a specific drug. It is preferably an abnormal acute dermal mucosal reaction. Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN), Stevens-Johnson syndrome, toxic epidermal necrolysis syndrome (SJS / TEN overlap syndrome) . However, it is not limited to the harmful reaction caused by lamotrigine in anticonvulsants.

As used herein, the term "biological sample" refers to all samples that can confirm the HLA-B genotype of the patient, preferably blood, plasma, serum, etc., but the presence or absence of HLA-B * The kind that can be confirmed is not limited thereto.

As used herein, the term " method for providing information on the prediction of adverse drug reaction "is a method for providing information on the risk that an adverse reaction due to a specific drug can be indicated. And to obtain information about the possibility of an adverse reaction to the drug.

As used herein, the term "kit " means a device for examining the presence or absence of HLA-B * 4403 and anticipating drug harmful reaction by anticonvulsants. B * 4403 can be confirmed by the presence of any limit. Preferably a probe or a primer set having a sequence complementary to the HLA-B * 4403 gene, or may be an antibody or an antibody specifically binding to HLA-B * 4403 It can be a form involving an aptamer. The term "probe or primer" refers to an oligonucleotide having a sequence complementary to HLA-B * 4403, preferably a portion of the sequence of exons 2 to 4 of HLA-B * 4403 But are not limited thereto.

The present invention provides a method for identifying HLA-B * 4403 from a biological sample isolated from a patient and providing information on prediction of adverse drug reaction by anticonvulsants. The anti-convulsant agent is preferably lamotrigine.

In one embodiment of the present invention, the method can further increase prediction accuracy by additionally identifying one or more clinical characteristics selected from the group consisting of patient's sex, liver dysfunction, and ocular disease. For example, a male with the HLA-B * 4403 genotype may have a higher severity of adverse reaction compared to women with the HLA-B * 4403 genotype. Abnormal liver function can be confirmed by measuring the concentration of aspartate aminotransferase (AST) or alanine aminotransferase (ALT) in the serum. The ophthalmic disease is corneal erosion, , Conjunctivitis, and decreased visual acuity (OCD).

In another embodiment of the present invention, the prediction is characterized by targeting a Korean person.

In another embodiment of the present invention, the method for identifying HLA-B * 4403 is not limited as long as it can identify a commonly used HLA-B genotype. Preferably, the HLA-B * 4403 is HLA microcytotoxicity test ), Flow cytometry, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR).

The present invention also provides a kit for predicting adverse drug reactions by anticonvulsants capable of confirming the presence of HLA-B * 4403. The kit comprises a probe having a sequence complementary to the HLA-B * 4403 gene, a probe or a set of primers and may also comprise an antibody or an aptamer specifically binding to HLA-B * 4403.

In the case of confirming the presence of HLA-B * 4403 from a biological sample isolated from a patient according to the present invention, the method of predicting drug harmful reaction by anticonvulsants can be performed by examining HLA- It is expected that the drug harmful reaction can be safely prevented at low cost and short term confirmation before the drug treatment because it can significantly reduce the suffering and / or the risk burden of the patient.

Brief Description of the Drawings Fig. 1 is a diagram showing the nucleotide sequences of Exons 2 to 4 of HLA-B * 4403 according to an embodiment of the present invention, wherein the sequence described in green is exon 2, the sequence described in red is exon 3, The sequence described means exon 4.

Hereinafter, the present invention will be described in more detail with reference to Examples. It will be apparent to those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example 1: SAMPLE ANALYSIS

For analysis of patients with Stevens-Johnson syndrome (SJS) and / or toxic epidermal necrolysis (TEN), patients previously diagnosed with Stevens-Johnson syndrome and / or toxic epidermal necrolysis The following factors were analyzed. The information of each patient is shown in Table 1. Patients with Stevens-Johnson syndrome who have less than 10% of the skin in which epidermal detachment is observed are those with Stevens-Johnson syndrome and toxic epidermal necrolysis, with 10-30% Were classified as toxic epidermal necrolysis, and the severity of the patients was classified using SCORTEN (SCORENE TOXIC EPIDERMAL NECROLSISIS), which is used as an indicator of severity of vesicular lesion disease. The SCORTEN level was measured by the age (> 40 years), pulse rate (> 120 times / min), presence of cancer or hematologic malignant neoplasms, overlap syndrome or toxic epidermal necrolysis, blood urea nitrogen (28 mg / dL), blood glucose concentration (> 252 mg / dL), and acidosis (bicarbonate <20 mEq / L) according to seven independent risk factors This is a numerical value indicating the severity of a vesicular lesion disease. If the SCORTEN value is 3 or more, it shows a mortality rate of 32% or more. The 30 patients were found to have SCORTEN level within 24 hours after admission to the hospital. The anemia (Hb <10 g / dL) was measured by measuring the concentration of hemoglobin for analysis of association with other organs. ([Cr]> 2.0 mg / L) by measuring the creatinine concentration in the serum and measuring the concentration of AST and ALT in the serum to detect liver function abnormality (AST or ALT > 50 IU / L), C-reactive protein (CRP,> 8 mg / L), and slit lamp examination, corneal erosion, conjunctivitis, The diagnosis of ocular complications such as decreased visual acuity was made. The HLA genotype of each patient was confirmed using the A LIFECODES antibody detection system (Luminex® platform). Sequences of exons 2 to 4 of HLA-B * 4403 were also amplified by polymerase chain reaction (PCR) to confirm the sequence. The sequence of exons 2 to 4 (SEQ ID NO: 1) of HLA-B * 4403 is shown in Fig. 1 (http://www.ebi.ac.uk/cgi-bin/ipd/imgt/hla/align .cgi).

For statistical analysis of the measured values, IBM SPSS Statistics 18.0 was used. In order to confirm the correlation between the HLA allele and the vulnerability of the drug, Fisher's exact test was used to confirm the relationship between the two groups. Pearson's validation method (Pearson? test), and interpreted as statistically significant for P-value <0.05.

Number Sex Age (yr) Hospital day phenotype SCORTEN Death One F 67 23 TEN 2 No 2 F 55 33 TEN 4 No 3 F 45 0 TEN 2 No 4 F 49 21 SJS 2 No 5 F 73 34 TEN 3 Yes 6 M 71 21 TEN 3 No 7 F 39 19 Overlap One No 8 F 77 4 SJS One No 9 F 52 16 SJS 3 No 10 M 74 24 Overlap 2 No 11 F 54 8 SJS One No 12 F 61 0 SJS No 13 F 74 9 SJS One No 14 M 39 10 SJS 0 No 15 F 42 9 SJS One No 16 M 61 19 SJS One No 17 M 72 0 SJS No 18 F 62 24 SJS 3 No 19 M 63 30 TEN 3 No 20 F 72 14 SJS 2 No 21 F 59 82 SJS 2 No 22 M 57 29 TEN 3 Yes 23 M 76 26 TEN 3 No 24 F 26 15 SJS One No 25 F 42 7 SJS One No 26 F 61 11 SJS 2 No 27 M 57 30 TEN 2 No 28 F 29 2 SJS 0 No 29 F 80 9 SJS 2 No 30 F 19 12 SJS 0 No

* IVIG: Intravenous immunoglobulin, F: Female M: Male, TEN: Toxic epidermal necrolysis, Overlap: Overlap syndrome, SJS: Stevens Johnson Syndrome

As shown in Table 1, the selected 30 patients were all Korean ethnically, with an average age of 56.9 ± 16.10 years and an average hospital stay of 18.0 ± 13.79 days. And 70% of the 30 patients were female and were more vulnerable to Stevens-Johnson syndrome and / or toxic epidermal necrolysis than females. Stevens-Johnson syndrome patients were composed of 19 patients (63.3%), Stevens-Johnson syndrome and toxic epidermal necrolysis patients (6.7%), and toxic epidermal necrolysis patients (9 patients) Respectively. Thirty patients were all treated with steroid therapy, and 13 of them (43.3%) received intravenous immunoglobulin (IVIG). The mean SCORTEN value of the surviving patients was 1.7 ± 1.04, and the mean SCORTEN value of the dead patients was 3.0 ± 0.00. The mean SCORTEN value of all patients was 1.8 ± 1.06, and the correlation between SCORTEN and mortality was P- value was 0.102, indicating no significance. The overall mortality rate was 6.6%.

Example 2: Stevens-Johnson syndrome and / or toxic epidermal necrolysis-induced drug analysis

In order to screen for the major causative agents causing Stevens-Johnson syndrome and / or toxic epidermal necrolysis, patients selected in Example 1 were treated with anticonvulsants, allopurinol, antibiotics, Drugs known to be causative agents, such as acetazolamide, acetaminophen, and herbal medication, are administered to identify the causative agent in each patient, and also to treat the patient with Stevens-Johnson syndrome / Or mean latency in which toxic epidermal necrolysis was developed were measured. The results are shown in Table 2.

Frequency (%) Mean latency (day) ± SD Mean age (yr) ± SD Anticonvulsants 26.7 36.00 ± 27.934 60.25 + 14.150 Allopurinol 26.7 19.50 ± 13.115 59.00 ± 15.764 Antibiotics 16.7 14.60 ± 11.415 64.63 ± 10.253 Acetazolamide 10 25.00 ± 8.660 62.67 ± 9.815 Acetaminophen 10 4.33 ± 3.055 34.00 ± 7.071 Herbs 6.7 6.50 7.778 34.67 ± 21.362 Etc 3.3 2 54.00 Sum 100 20.14 ± 18.969 56.93 ± 16.099

As shown in Table 2, 26.7% of cases of anticonvulsant or allopurinol, 16.7% of cases of antibiotics, 10% of cases of acetic acid amide or acetaminophen, respectively, Was 6.7% and 3.3% when no causative drug was identified. It was confirmed that the main drug causing Stevens-Johnson syndrome and / or toxic epidermal necrolysis in Koreans was an anticonvulsant and / or allopurinol. In addition, the latency period was different depending on the drug. In the case of anticonvulsants, the average latency was 36.00 ± 27.934 days, and in the case of acetaminophen, the mean was 4.33 ± 3.055 days. Period. In the case of anticonvulsants, allopurinol, antibiotics, and acetic acid amide, the average age was 59 to 65 years old while the acetaminophen and herbal medicines were 34 years old.

Example 3: Analysis of association with other organizations

In order to determine whether the Stevens-Johnson syndrome and / or toxic epidermal necrolysis was related to the deterioration of other organs, the dysfunction of other organs was measured in the same manner as in Example 1. The results are shown in Table 3.

Frequency (%) Older age than 40 years old 83.3 Tachycardia (HR > 120 beats / minute) 16.7 Comorbid malignancy 10.0 BUN> 28 mg / dL 20.0 Glucose> 252 mg / dL 3.3 Bicarbonate <20 mEq / L 10.0 CRP elevation (> 8 mg / L) 76.7 Anemia (Hb < 10 g / dL) 30.0 Kidney injury (Creatinine> 2 mg / dL) 16.7 Liver injury (AST or ALT> 50 IU / L) 63.3 Ocular lesion 63.3

CRP: C-reactive protein; HR: Heart rate; BUN: Blood urea nitrogen

As shown in Table 3, 83.3% of all patients were over 40 years of age. The risk of SCORTEN, an indicator of the severity of bullous disease, was high in heart rate (tachycardia), cancer or hematologic malignant neoplasm (BUN), high concentration of glucose in the blood, and acidosis were found to be correlated with the presence of organisms, blood urea nitrogen (BUN). On the other hand, 76.7% of the patients with C-reactive protein increased acute inflammatory response, 63.3% with liver dysfunction, and 63.3% with ocular complications. Respectively. The above results indicate that Korean Stevens-Johnson syndrome and / or toxic epidermal necrolysis are highly correlated with age, C-reactive protein, liver dysfunction, and ocular complications.

Example 4: Analysis of association with sex

Stevens-Johnson syndrome and / or toxic epidermal necrolysis were analyzed for association with gender. The results are shown in Table 4.

SJS Overlap TEN Female 16 (76.2%) 1 (4.8%) 4 (19.0%) Male 3 (33.3%) 1 (11.1%) 5 (55.6%)

As shown in Table 4, in the case of women, the combination of Stevenson-Johnson syndrome and toxic epidermal necrolysis or the toxic epidermal necrolysis was only 23.8%, while for men, 66.7% (P <0.05). In addition, it was confirmed that the patients were caused by the above patients. These results show that Korean men have more severe symptoms of bulbar disease than women.

Example 5: Analysis of association with HLA-B alleles

The HLA genotypes were measured in the same manner as in Example 1 to confirm whether the signs of Stevens-Johnson syndrome and / or toxic epidermal necrolysis were associated with HLA-B alleles. Of the 30 patients, 18 had HLA-B expression, and the HLA-B genotypes were identified in each patient. The results are shown in Table 5.

Number Culpric drug HLA-B genotype One Lamotrigine B * 44: 03, B * 52: 01 2 Lamotrigine B * 44: 02, B * 44: 03 3 Lamotrigine B * 44: 03, B * 51: 01 4 Lamotrigine B * 15: 07, B * 39: 01 5 Lamotrigine B * 15: 01, B * 35: 03 6 Allopurinol B * 35, B * 58 7 Allopurinol B * 44: 02, B * 58: 01 8 Allopurinol B * 07: 02, B * 58: 01 9 Allopurinol B * 15: 01, B * 40: 06 10 Allopurinol B * 40: 06, B * 5801 11 Allopurinol B * 15: 11, B * 58: 01 12 Allopurinol B * 08: 01, B * 58: 01 13 Acetazolamide B * 07: 02, B * 59: 01 14 Acetazolamide B * 40: 06, B * 59: 01 15 Herbal B * 15: 01, B * 44: 03 16 Acetaminophen B * 40: 01, B * 58: 01 17 amoxacillin B * 08: 01, B * 51: 01 18 unknown B * 07: 02, B * 44: 03

As shown in Table 5, HLA-B * 4403 alleles were expressed in three of five patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by lamotrigine, one of the anticonvulsant drugs , Whereas no patient was found to express HLA-B * 1502 alleles or HLA-B * 3101 alleles. Lamotrigine is a recently developed anticonvulsant drug that is increasingly being used as an alternative to carbamazepine, an anticonvulsant drug that is commonly used. The HLA-B * 5801 allele was expressed in 5 of 7 patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by allopurinol, and in 2 patients, HLA-B * 4006 allele And all two patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by acetazolamide were found to express the HLA-B * 5901 allele.

As shown in Table 5, HLA-B * 4403 alleles were expressed in three of five patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by lamotrigine, one of the anticonvulsant drugs , Whereas no patient was found to express HLA-B * 1502 alleles or HLA-B * 3101 alleles. Lamotrigine is a recently developed anticonvulsant drug that is increasingly being used as an alternative to carbamazepine, an anticonvulsant drug that is commonly used. The HLA-B * 5801 allele was expressed in 5 of 7 patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by allopurinol, and in 2 patients, HLA-B * 4006 allele And all two patients with Stevens-Johnson syndrome and / or toxic epidermal necrolysis caused by acetazolamide were found to express the HLA-B * 5901 allele.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

<110> Industry-Academic Cooperation Foundation, Yonsei University <120> Method of providing the information for predicting adverse drug          reaction <130> DPB140046 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1089 <212> DNA <213> Homo sapiens <400> 1 atgcgggtca cggcgccccg aaccctcctc ctgctgctct ggggggcagt ggccctgacc 60 gagacctggg ccggctccca ctccatgagg tatttctaca ccgccatgtc ccggcccggc 120 cgcggggagc cccgcttcat caccgtgggc tacgtggacg acacgctgtt cgtgaggttc 180 gacycgacg ccacgagtcc gaggaaggag ccgcgggcgc catggataga gcaggagggg 240 ccggagtatt gggaccggga gacacagatc tccaagacca acacacagac ttaccgagag 300 aacctgcgca ccgcgctccg ctactacaac cagagcgagg ccgggtctca catcatccag 360 aggatgtacg gctgcgacgt ggggccggac gggcgcctcc tccgcgggta tgaccaggac 420 gcctacgacg gcaaggatta catcgccctg aacgaggacc tgagctcctg gaccgcggcg 480 gacaccgcgg ctcagatcac ccagcgcaag tgggaggcgg cccgtgtggc ggagcagctg 540 agagcctacc tggagggcct gtgcgtggag tcgctccgca gatacctgga gaacgggaag 600 gagacgctgc agcgcgcgga ccccccaaag acacatgtga cccaccaccc catctctgac 660 catgaggtca ccctgaggtg ctgggccctg ggcttctacc ctgcggagat cacactgacc 720 tggcagcggg atggcgagga ccaaactcag gacaccgagc ttgtggagac cagaccagca 780 ggagatagaa ccttccagaa gtgggcagct gtggtggtgc cttctggaga agagcagaga 840 tacacatgcc atgtacagca tgaggggctg ccgaagcccc tcaccctgag atgggagccg 900 tcttcccagt ccaccgtccc catcgtgggc attgttgctg gcctggctgt cctagcagtt 960 gtggtcatcg gagctgtggt cgctgctgtg atgtgtagga ggaagagctc aggtggaaaa 1020 ggagggagct actctcaggc tgcgtgcagc gacagtgccc agggctctga tgtgtctctc 1080 acagcttga 1089

Claims (15)

Methods for identifying HLA-B * 4403 from biological samples isolated from Korean patients and providing information on the prediction of drug adverse events by lamotrigine. The method according to claim 1,
Wherein the method additionally identifies one or more clinical characteristics selected from the group consisting of a patient's sex, liver dysfunction, and ocular disease. 3. The method of claim 2,
Wherein the liver dysfunction is characterized by measuring the concentration of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the serum. 3. The method of claim 2,
Wherein the ophthalmic disease is at least one selected from the group consisting of corneal erosion, conjunctivitis and decreased visual acuity. delete The method according to claim 1,
The adverse drug reaction may be Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN) or Stevens-Johnson syndrome-toxic epidermal necrolysis syndrome (SJS / TEN overlap syndrome) &Lt; / RTI &gt; delete The method according to claim 1,
Wherein said biological sample is blood, plasma or serum. The method according to claim 1,
The HLA-B * 4403 can be identified by HLA microcytotoxicity test (MCT), flow cytometry, enzyme-linked immunosorbent assay (ELISA) or Characterized in that it is a polymerase chain reaction (PCR). A kit for predicting drug adverse reactions by a Korean lamotrigine comprising a probe or a set of primers having a sequence complementary to the HLA-B * 4403 gene.
delete 11. The method of claim 10,
The adverse drug reaction may be Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN) or Stevens-Johnson syndrome-toxic epidermal necrolysis syndrome (SJS / TEN overlap syndrome) Features a kit. A kit for predicting adverse drug reaction by lamotrigine in Korean, comprising an antibody or an aptamer specifically binding to HLA-B * 4403.
delete 14. The method of claim 13,
The adverse drug reaction may be Stevens-Johnson syndrome (SJS), toxic epidermal necrolysis (TEN) or Stevens-Johnson syndrome-toxic epidermal necrolysis syndrome (SJS / TEN overlap syndrome) Features a kit.

Images