KR20170109813A

KR20170109813A - A biomarker for predicting reactivity with sorafenib and use of the same

Info

Publication number: KR20170109813A
Application number: KR1020160033879A
Authority: KR
Inventors: 김수옥; 김석준; 홍선표; 홍성수; 조우재; 이승화
Original assignee: (주)진매트릭스
Priority date: 2016-03-22
Filing date: 2016-03-22
Publication date: 2017-10-10

Abstract

The present invention relates to genetic markers for predicting the sorapenib response in patients with hepatocellular carcinoma using genetic polymorphisms, and more particularly to genetic markers for predicting the response to sorafenib evaluated by lesions in hepatocarcinoma patients, Compositions and applications thereof. The genetic marker analysis can be used to perform individualized chemotherapy of hepatocellular carcinoma patients.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a genetic marker for predicting the sorapenib response in hepatocellular carcinoma patients,

The present invention relates to genetic markers for predicting the sorapenib response in hepatocarcinoma patients using gene polymorphisms and their use. More specifically, the present invention relates to a method for predicting reactivity to sorapenib by identifying a specific single nucleotide polymorphism (SNP) base having a significant correlation with the response to sorapenib anticancer agent in a hepatocellular carcinoma patient, The present invention relates to a composition for predicting the response and survival prognosis of a patient suffering from hepatocellular carcinoma including polynucleotide, polypeptide or cDNA thereof, and a gene analysis kit comprising the same.

Hepatocellular carcinoma is a malignant tumor that is the third most common cancer death in the world. 70-90% of patients with hepatocellular carcinoma are accompanied by cirrhosis or chronic liver disease. The causative factors of hepatocellular carcinoma are relatively well known and include infection with chronic hepatitis B virus (HBV), infection with chronic hepatitis C virus (HCV), non-alcoholic (Et al. Gastroenterology 2007; 132: 1144-1152) (Poon et al., The lancetoncology (2006)), 2009; 10: 1111-1118). In particular, the incidence of hepatocellular carcinoma (HCC) is increasing steadily in the Asian region, including Korea, where hepatitis virus is highly prevalent, and is the second most common cause of cancer deaths (Poon et al. -1118).

More than 60% of hepatocellular carcinoma patients are diagnosed as advanced hepatocellular carcinoma and do not attempt curative treatment. In patients who can not be treated with curative treatment, various treatments including transarterial chemoembolization (TACE) are tried. In particular, in the case of systemic chemotherapy, sorafenib was reported to improve the overall survival rate of patients with hepatocellular carcinoma in a randomized, triple-phase study in 2008 (Liovet et al., New England Journal of Medicine 2008; 359: 378-390) .

However, there is a limit to the reduction in the size of tumors in patients treated with sorafenib, which is only 3.3%, and the increase in survival remains within 2-3 months (Liovet et al., New England Journal of Medicine 2008; 359 : 378-390) (Cheng et al., The lancet oncology 2009 10.1; 25-34). Despite the high incidence of hepatocellular carcinoma, the incidence of hepatocellular carcinoma is still low, and the incidence of hepatocellular carcinoma is increasing. Therefore, a number of research institutes are studying methods and markers for predicting the response of sorapenib. However, there is still a lack of methods for predicting the response to sorapenib, resulting in waste of treatment costs and inconvenience for patients with hepatocellular carcinoma.

There is a disadvantage in that it is not easy to study because the overall response rate (ORR) of sorapenib in hepatocellular carcinoma is low compared to the chemotherapy of other carcinomas (Liovet et al., New England Journal of Medicine 2008; 359: 378-390 ). In addition, there is a strong correlation between the overall survival (OS), progression-free survival (PFS), disease-free survival (DFS), time to tumor progression (TTP), as described above, has a disadvantage in that it is difficult to evaluate the prediction method and the predictive marker since the survival time is only 2 to 3 months (Shao et al., World Journal of Gastroenterology 2015; 21: 10336-10347).

The object of the present invention is to solve the above-mentioned problems and to provide a novel SNP that has a significant correlation with the prediction of reactivity to sorapenib in gene samples collected from patients with hepatocellular carcinoma By identifying the bases, it is intended to provide a genetic marker that predicts the response and prognosis of sorapenib in patients with hepatocellular carcinoma.

Another object of the present invention is to provide a kit for predicting the responsiveness to sorapenib in a hepatocellular carcinoma patient comprising a polynucleotide capable of confirming SNP markers specified in the present invention.

In order to achieve the above object, the present invention provides a method for identifying a single nucleotide polymorphism (SNP) of the 26th base sequence of SEQ ID NO: 1 in a gene sample obtained from a patient with hepatocellular carcinoma, A or G / G genotype, Sorafenib is more likely to respond to sorafenib than G / A genotype, and Sorafenib G / A or G / The present invention provides a method for providing information for predicting the response and survival prospect of Soraphenib in a patient with hepatocellular carcinoma.

In addition, the present invention confirms the single nucleotide polymorphism (SNP) of the 26th base of the sequence shown in SEQ ID NO: 1 in a gene sample obtained from a malignant neoplasm patient, and confirms that a G / A or G / And G / G genotypes were similar to those of patients with A / A genotypes compared to patients with A / A genotypes. The present invention provides a method for providing information for predicting the reactivity and survival prognosis of chemotherapy with the same mechanism as that of sorapenib in a malignant neoplasm patient.

In one embodiment of the present invention, the chemotherapy regimen with the same mechanism as sorapenib is preferably Regorafenib, Lenvatinib, Donafenib, or Brivanib But not limited to this.

The present invention also relates to a method of treating a patient suffering from hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of SEQ ID NO: 1 or a complementary polynucleotide thereof, Based on the total weight of the composition.

The present invention also relates to a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a hepatocyte cancer cell comprising a primer pair or probe for its complementary polynucleotide as an essential component And a composition for predicting the response and survival prognosis of the patient to sorapenib.

In one embodiment of the present invention, the primer pair sequence is preferably composed of the nucleotide sequences of SEQ ID NOS: 3 to 4 or SEQ ID NOS: 7 to 8, but is not limited thereto.

In another embodiment of the present invention, the probe sequence is preferably composed of the nucleotide sequence of SEQ ID NO: 9 or 10, but is not limited thereto.

The present invention also relates to a method of treating a patient suffering from hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of SEQ ID NO: 1 or a complementary polynucleotide thereof, Based on the predicted value.

The present invention also relates to a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a hepatocyte cancer cell comprising a primer pair or probe for its complementary polynucleotide as an essential component A kit for predicting the response and survival prognosis of patients with Sorapenib.

In addition, the present invention confirms a single base polymorphism (SNP) of the 26th base of the sequence shown in SEQ ID NO: 2 in a gene sample obtained from a patient with hepatocellular carcinoma, and when the C / C genotype and the G / , And predicts that it will not respond to sorafenib when the C / C genotype and the G / G genotype are compared with the C / C genotype when the C / G genotype is present A method for providing information for predicting the response and survival prognosis of sorapenib in patients with hepatocellular carcinoma.

In addition, the present invention confirms a single base polymorphism (SNP) of the 26th base of the sequence shown in SEQ ID NO: 2 in a gene sample obtained from a malignant neoplasm patient, and when the C / C genotype and the G / G / G genotype in the presence of C / C genotypes and G / G genotypes in the presence of C / G genotypes compared to that of sorafenib The present invention provides a method of providing information for predicting the reactivity and survival prognosis of chemotherapy with the same mechanism as that of sorafenib in a malignant neoplasm patient, which is predicted not to respond to chemotherapy with a history of cancer.

The present invention also relates to the use of a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof, to a patient suffering from hepatocellular carcinoma, Based on the total weight of the composition.

The present invention also relates to a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence shown in SEQ ID NO: 2 or hepatocellular carcinoma cells comprising a primer pair or probe for its complementary polynucleotide as an essential component And a composition for predicting the response and survival prognosis of the patient to sorapenib.

In one embodiment of the present invention, the primer pair sequence is preferably composed of the nucleotide sequence of SEQ ID NO: 5 to 6 or SEQ ID NO: 11 to 12, but is not limited thereto.

In another embodiment of the present invention, the probe sequence is preferably composed of the nucleotide sequence of SEQ ID NO: 13 or 14, but is not limited thereto.

The present invention also relates to a method of treating a patient suffering from hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof, Based on the predicted value.

The present invention also relates to a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a hepatocyte cancer cell comprising a primer pair or probe for the complementary polynucleotide thereof as an essential component A kit for predicting the response and survival prognosis of patients with Sorapenib.

In one embodiment of the present invention, the primer pair sequence consists of the nucleotide sequence of SEQ ID NO: 5 to 6 or SEQ ID NO: 11 to 12, and the probe sequence comprises the nucleotide sequence of SEQ ID NO: 13 or 14, It is not limited.

Hereinafter, the present invention will be described.

The present inventors have evaluated a patient with hepatocellular carcinoma whose response has been evaluated by the revised RECIST guideline v1.1 (EI et al., European Journal of Cancer 2009; 45: 228-247) A gene marker capable of predicting the sorapenib response was found, and the present invention was completed.

The present invention relates to a gene sample obtained from a hepatocellular carcinoma patient, which comprises a base of a SNP (NCBI refSNP ID: rs7923609) located in the 26th base of SEQ ID NO: 1 and a base of 26th base of SEQ ID NO: 2 SNP (NCBI refSNP ID: rs2910164) is identified to provide a genetic marker that predicts sorapenib reactivity.

The sequence of the gene marker is shown in Table 1 below. The refSNP ID of NCBI for each SNP in Table 1 indicates the sequence of the SNP and its position. Those skilled in the art can easily identify the position and sequence of the SNP using the above numbers. It will be apparent to those skilled in the art that the specific sequence corresponding to the refSNP ID of the SNPs registered in the NCBI may be varied slightly depending on the results of the study of the succeeding genes and such altered sequences are also included within the scope of the present invention.

In the present invention, the rs7923609 A / A genotype is associated with a non-responsiveness to sorapenib. Therefore, when the rs7923609 A / A genotype is detected in hepatocellular carcinoma patients, the reactivity to sorapenib is considered to be low and the G / A genotype When the G / G genotype is detected, sorapenib reactivity is considered high.

Because rs2910164 C / C genotype and G / G genotype are related to reactivity to sorapenib, when rs2910164 C / C genotype and G / G genotype are detected in hepatocellular carcinoma patients, When the / G genotype is detected, reactivity to sorapenib is considered low.

In particular, according to Example 4 described later, patients with hepatocellular carcinoma having the A / A genotype of rs7923609 are 3.06 times more likely to not respond to sorapenib than patients with hepatocellular carcinoma having G / A or G / G genotype (P = 0.0097, OR = 3.06, CI = 1.26-7.38). In addition, patients with hepatocellular carcinoma with G / A or G / G genotype of rs7923609 are 3.06 times more likely to respond to sorafenib than patients with hepatocellular carcinoma with A / A genotype (P = 0.0097, OR = 3.06, CI 1.26-7.38)

rs2910164 Patients with hepatocellular carcinoma with C / C genotype and G / G genotype are 2.26 times more likely to respond to sorafenib than those with C / G genotype (P = 0.044, OR = 2.26, CI 1.01-5.04). Patients with hepatocellular carcinoma of the C / G genotype are 2.26 times more likely to be unresponsive to sorapenib than those with the C / C genotype and the G / G genotype (P = 0.044, OR = 2.26, CI 1.01-5.04)

Also, according to Tables 7 and 10 of Example 4 described later, when the clinical information was corrected by covariate, the rs7923609 marker and the rs2910164 marker were significantly correlated with the sorapenib response. That is, the SNP analyzed by the present invention from hepatocellular carcinoma patients may be related to the sorapenib response irrespective of clinical information.

In particular, the SNP of the present invention can be used as a biomarker for predicting the response before treatment with sorapenib. Therefore, the genetic marker analysis of the present invention will be helpful in the screening of small groups that are effective in the treatment of sorapenib, and may be helpful in determining the treatment of sorapenib in patients with hepatocellular carcinoma. These results show that the SNP identified in the present invention can be applied as a genetic marker capable of predicting the response of sorapenib.

In the present invention, the patient refers to a patient suffering from hepatocellular carcinoma. A gene sample is obtained from tissues, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid, urine and the like obtained from these patients with hepatocellular carcinoma. The gene sample includes cDNA synthesized from DNA, mRNA or mRNA.

In the present invention, the predictive response to sorafenib was evaluated based on the revised RECIST guideline according to malignant neoplasia, recurrence, metastatic spread, tumor size and progression in HCC patients. Thus, the present invention allows predicting the response based on a revised RECIST guideline after treatment with sorapenib.

The present invention also provides a composition for predicting the reactivity and survival prognosis of Sorapanib in hepatocellular carcinoma patients to achieve the above-mentioned other objects.

The SNP (NCBI refSNP ID: rs7923609) located in the 26th nucleotide of SEQ ID NO: 1 and the SNP (NCBI refSNP ID: rs7923609) located in the 26th nucleotide of SEQ ID NO: lt; RTI ID = 0.0 > rs2910164) < / RTI >

The polynucleotide or its complementary polynucleotide according to the present invention is a polymorphic sequence. A polymorphic sequence refers to a sequence comprising a polymorphic site representing a single base polymorphism in the nucleotide sequence. In the present invention, the polynucleotide may be DNA or RNA.

In the present invention, the allele-specific polynucleotide may be an allele-specific primer. The appropriate length of the primer may vary depending on the purpose of use, but it is generally composed of 30 bases of 15 cycles. The primer sequence need not be completely complementary to the template, but should be sufficiently complementary to hybridize with the template. The primer hybridizes to a DNA sequence containing a polymorphic site to amplify a DNA fragment containing the polymorphic site. The primer of the present invention can be used for a diagnostic kit such as a real time polymerase chain reaction (PCR), a real time gene amplification (real time polymerase chain reaction) and the like for detecting alleles and predicting reactivity to sorapenib in patients with hepatocellular carcinoma.

The kits described above in the present invention may include polynucleotides, polypeptides, cDNAs and the like of the present invention as well as other component compositions, solutions or devices of a kind or more suitable for the assay method. In one embodiment, the kit of the present invention may be a kit containing the necessary elements necessary to perform PCR, and may be a test tube or other suitable container, a reaction buffer (pH and magnesium concentrations vary), deoxynucleotides (dNTPs) Enzymes such as Taq polymerase and reverse transcriptase, DNase, RNAse inhibitor, DEPC-water and sterile water.

Identification of the genotypes of the SNPs of the present invention can be carried out by sequencing analysis, sanger sequencing using an automatic nucleotide sequencer, pyrosequencing, next generation sequencing, hybridization with a microarray, PCR- Restriction fragment length polymorphism (RELP), single strand conformation polymorphism (PCR), specific sequence oligonucleotides (PCR-SSO), allele specific oligonucleotide (ASO) hybridization in combination with PCR-SSO and dot hybridization, A known method such as PCR, MALDI-TOF / MS, rolling circle amplification, HRM (high resolution melting) real time gene amplification, primer extension, Southern blot hybridization and dot hybridization . Further, the SNP polymorphism results can be statistically analyzed using statistical analysis methods commonly used in the art. For example, continuous variables such as Chi-square test, linear regression line analysis, and multiple logistic regression analysis, categorical variables, variables, odds ratios, and 95% confidence intervals.

Since the gene markers identified in the present invention can be applied as biomarkers capable of predicting the response and survival rate of sorapenib anticancer agent, analysis of the gene markers of the present invention can be used to select small groups effective for sorapenib chemotherapy, And can be usefully used to determine the therapeutic treatment of cancer patients.

Fig. 1 is a primer sequence of PCR of rs7923609 and rs2910164 in the gene analysis performed in Example 2. Fig.
Fig. 2 is a primer and probe sequence of the Taqman assay of rs7923609 and rs2910164 in the gene analysis performed in Example 2. Fig.
Fig. 3 is an example of an allele discrimination plot of the Taqman assay of rs7923609 SNP in the gene analysis performed in Example 2. Fig.
Figure 4 is an example of an allele discrimination plot of the Taqman assay of the rs2910164 SNP in the gene analysis performed in Example 2. Fig.

Hereinafter, the present invention will be described in detail with reference to examples. It should be understood, however, that the following examples of the present invention are for the purpose of illustrating the present invention and do not limit or limit the scope of the present invention. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Unless otherwise indicated, nucleic acids are written in 5'-3 'orientation from left to right.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice to test the present invention, the preferred materials and methods are described herein.

Example 1. Patient collection and sample preparation

1) Target patient

From August 2013 to August 2014, patients with hepatocellular carcinoma scheduled for treatment with sorafenib were recruited randomly from the Department of Internal Medicine, Seoul National University Hospital. This experiment was conducted under the permission of Seoul National University Medical Research Ethics Review Committee, and the patient 's written consent was obtained based on the doctor' s notice.

2) Clinical data collection

Blood was collected from the patients and serum and buffy coat were separated. Serum was analyzed by alpha-fetoprotein (AFP) and PIVKA-II (protein induced by vitamin K absence II) biochemical tests, and the patient's age, sex, etiology, lymph node metastasis and distant metastasis were summarized. The clinical profile for 119 patients is shown in Table 2.

The progression of hepatocellular carcinoma in the patients was classified according to the TNM stage (TNM Classification of Malignant Tumors) and BCLC (Barcelona clinic liver cancer) stage, and the information was summarized. Table 3 shows the hepatocellular carcinoma status profile for 119 patients.

The evaluation of sorafenib response was evaluated by CR (complete response), PR (partial response), SD (stable disease) and PD (progressive disease) by the revised RECIST guideline. 4. Patients who could not be judged were excluded from the statistical analysis of Example 3 described later.

3) DNA extraction

DNA was extracted from 119 buffy coat samples collected from the patients using a QIAamp DNA Blood Mini Kit 250 (Qiagen, Germany) based on the kit manual.

Example 2. Genetic polymorphism screening and genotyping

(AFP, BCHE, LDH, ALP, GGT, ALT, and PIVKA) that are known to be related to the concentration of various hepatocellular carcinoma markers (AFP, BCHE, ALT, and PIVKA) in order to predict the reactivity of hepatocarcinoma patients to anticancer agents through public database and literature search. And 19 SNPs were selected. We also investigated the gene and metabolism related genes of sorafenib. Twelve SNPs with minor allele frequencies of 10% or more were identified in the genes. Four SNPs were also selected for the microRNAs associated with sorapenib response and expression levels. A total of 35 SNPs were genotyped using an automated sequencing system (ABI, 3130xl). For the quality control of the experiment, the genotype was analyzed without informing the patient, and both the forward and reverse analyzes were performed more than once. In addition, 10% of the analyzed samples were randomly selected and revalidated using an automated sequencer. In addition, genotype analysis results were verified by Taqman assay using real-time PCR (Real-time PCR) for genetic markers with statistically significant significance.

Example 3. Statistical Analysis

Whether or not the genotypes of hepatocellular carcinoma patients analyzed in the present invention were in a Hardy-Weinberg (HWE) equilibrium was tested using a degree of freedom for goodness-of-fit χ2 test . Patients who were treated with sorafenib were evaluated as RECIST based on revised RECIST, and classified into CR (complete response), PR (partial response) and SD (stable disease) Non-responders. Genotype comparison between the response group and the non-response group The correlation analysis used the χ2 tests for the categorical variables. Also, the association between genotypes and sorapenib responses was tested in 95% confidence interval (OR), odds ratios (OR), and multiple logistic regression analysis was used to measure age, sex, , Lymph node metastasis, distant metastasis, BCLS stage, and covariate of TNM stage. In addition, genomic analysis was carried out according to the allele genotype by codominant model, dominant model, recessive model and overdominant model.

Example 4. Results of genetic polymorphism analysis

The rs7923609 marker and rs2910164 of the 35 SNPs analyzed through the procedures of Example 2 and Example 3 showed statistically significant results.

Table 5 summarizes analyzed genotypes of rs7923609 markers. In Table 5, the genotypic distribution of the rs7923609 marker of the present invention showed a deviation in the Hein-Weinberg equilibrium (HWE) in the unreacted group (P = 0.019) but the deviation of the reaction group was not significant (P = 0.973). These deviations suggest a bias in the unresponsive specimen, which may be additional evidence of the association of the rs7923609 marker with the unreacted group. The genotypes analyzed showed a significant difference (P = 0.035) between the response group and the non-response group, and a significant difference (P = 0.025) between the response group and the no response group was found in each allelic frequency .

Table 6 shows the results of analyzing the ratios of the rs7923609 markers for dominance, dominance, temperament and dominance. The significance of the higher marker in the dominant model was confirmed (P = 0.0097). The results of the nonparametric analysis show that the chance of not responding to sorapenib is about 3.06 times higher in patients with hepatocellular carcinoma with A / A genotype than those with G / A genotype and G / G genotype (P = 0.0097 OR 3.06, CI 1.26-7.38).

In addition, the analysis of the above results shows that patients with hepatocellular carcinoma with G / A genotype and G / G genotype are 3.06 times more likely to respond to sorapenib than patients with A / A genotype (P = 0.0097, OR = 3.06, CI 1.26-7.38).

Table 7 shows the analysis of the analyzed genotypes of rs7923609 by applying covariance, dominance, dominance, and dominance models by multivariate logistic regression analysis by covariance of age, sex, etiology, lymph node metastasis, and distant metastasis. (P = 0.0035, OR 3.76, CI 1.48-9.55), indicating that the association of the rs7923609 marker with the sorapenib response was not due to clinical variables. (P = 0.023, OR = 2.93, CI = 1.12-7.66) was also observed when the sex, age, and BLCL stage were corrected for covariates in addition to the results of Table 7, . In addition, a significant correlation was found in the dominant model when sex, age, and TNM stages were corrected (P = 0.034, OR 2.86, CI 1.05-7.79).

Table 8 summarizes analyzed genotypes of the rs2910164 marker. In Table 7, the genotypic distribution of the rs2910164 marker of the present invention showed no deviation (P> 0.05) between the sorapenib reaction group (P = 0.056) and the non-reacted group with the Hein-Weinberg Equilibrium (HWE). There was no statistically significant difference in genotype frequencies and allele frequencies (P> 0.05).

Table 9 shows the result of analyzing the correspondence ratio of the rs2910164 marker by the gene model. As shown in Table 8, there was no statistical significance in the boreal model. However, the significance of the marker was confirmed in the hyperpolarity model (P = 0.044). In addition, the non-response analysis indicates that patients with C / C and G / G genotypes are 2.26 times more likely to respond to sorapenib than patients with C / G genotype (P = 0.044, OR 2.26, CI 1.01-5.04).

In addition, the results of the above analysis can be interpreted differently, suggesting that patients with hepatocellular carcinoma with C / G genotype are 2.26 times more likely to not respond to sorapenib than those with hepatocellular carcinoma with C / C genotype and G / G genotype (P = 0.044, OR = 2.26, CI 1.01-5.04).

Table 10 summarizes the frequency, dominance, dominance, and dominance models of the gene frequencies and correspondence tables for each model in Table 9 by multivariate logistic regression analysis by covariance of age, sex, etiology, lymph node metastasis, and distant metastasis Respectively. (P = 0.01, OR 3.08, CI 1.27-7.48), indicating that the association of the rs2910164 marker with the sorapenib response was not due to clinical variables. (P = 0.039, OR 2.61, CI 1.02-6.66) when the sex, age, and TNM stage were corrected for covariates in addition to the results of Table 10, which were not described in detail in the present invention, .

While the invention has been shown and described with reference to certain preferred embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention. It will be appreciated that such modifications and variations are also contemplated by the present invention.

<110> GENEMATRIX INC. <120> A BIOMARKER FOR PREDICTING REACTIVITY WITH SORAFENIB AND USE OF THE SAME <130> P16-0018 <160> 14 <170> Kopatentin 2.0 <210> 1 <211> 51 <212> DNA <213> Homo sapiens <400> 1 ctactggaat taatgacaat atcccattgc atagacttta aaataaacat a 51 <210> 2 <211> 51 <212> DNA <213> Homo sapiens <400> 2 catgggttgt gtcagtgtca gacctgtgaa attcagttct tcagctggga t 51 <210> 3 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 tcgttgtttc aatttgcatt cc 22 <210> 4 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 caaacaagca ttctcacttc acttg 25 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 ggaagcagct gcattggatt 20 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 tctgccttct gtctccagtc t 21 <210> 7 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 7 tcatagtttg cttagccagt tg 22 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 8 agcacacata acctgactac 20 <210> 9 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> probe <400> 9 atgacaatat cccattgca 19 <210> 10 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> probe <400> 10 atgacaatat cccgttgca 19 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 11 caagcccacg atgacagaga 20 <210> 12 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 12 gagaactgaa ttccatgggt tgtg 24 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> probe <400> 13 ctgaatttca caggtctga 19 <210> 14 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> probe <400> 14 aactgaattt cagaggtctg a 21

Claims

Single nucleotide polymorphisms (SNPs) of the 26th base of the sequence shown in SEQ ID NO: 1 were confirmed in a gene sample obtained from patients with hepatocellular carcinoma, and in case of having A / A genotype and G / A or G / G genotype (Sorafenib), and it is predicted that it will not respond to Sorafenib,
To predict sorafenib reactivity and survival prognosis in hepatocellular carcinoma patients with G / A or G / G genotypes, which are predicted to respond to Sorafenib compared to patients with A / A genotype How to provide information.

Single nucleotide polymorphisms (SNPs) of the 26th base of the sequence shown in SEQ ID NO: 1 were confirmed in a gene sample obtained from a malignant neoplasm patient, and those having the A / A genotype and the G / A or G / G genotype . In addition, it is predicted not to respond to anticancer chemotherapy with the same mechanism as sorapenib,
It is similar to sorafenib in patients with malignant neoplasm characterized by the anticipation that patients with G / A or G / G genotypes will respond to chemotherapy with the same mechanism as sorapenib compared with those with A / A genotype Methods for providing information for predicting the reactivity and survival prognosis of chemotherapeutic chemotherapy.

3. The method according to claim 2, wherein the chemotherapy regimen having the same mechanism as that of sorapenib is Regorafenib, Lenvatinib, Donafenib, or Brivanib. .

A composition for predicting the reactivity and survival prognosis of sorapenib in patients with hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a complementary polynucleotide thereof .

1 or a complementary polynucleotide consisting of 10 or more contiguous bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a complementary polynucleotide thereof as an essential component, For the prediction of reactivity and survival prognosis.

6. The composition according to claim 5, wherein the primer pair sequence comprises the nucleotide sequences of SEQ ID NOS: 3 to 4 or SEQ ID NOS: 7 to 8.

6. The composition according to claim 5, wherein the probe sequence comprises the nucleotide sequence of SEQ ID NO: 9 or 10. 10. A composition for predicting the response and survival prognosis of Sorapenib in patients with hepatocellular carcinoma.

A kit for predicting the response and survival prognosis of sorapenib in patients with hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a complementary polynucleotide thereof .

1 or a complementary polynucleotide consisting of 10 or more contiguous bases comprising the 26th base of the sequence represented by SEQ ID NO: 1 or a complementary polynucleotide thereof as an essential component, Kits for the prediction of reactivity and survival prognosis.

10. The method according to claim 9, wherein the primer pair sequence comprises the nucleotide sequence of SEQ ID NO: 3 to 4 or SEQ ID NO: 7 to 8, and the probe sequence comprises the nucleotide sequence of SEQ ID NO: Kit for prediction of reactivity and survival prognosis for sorafenib.

When the single nucleotide polymorphism (SNP) of the 26th base of the sequence shown in SEQ ID NO: 2 is confirmed in a gene sample obtained from a patient with hepatocellular carcinoma and the C / C genotype and the G / G genotype have a C / G genotype , And it is predicted that it will respond to sorafenib,
The present invention provides a method for predicting the response and survival prognosis of sorapenib in patients with hepatocellular carcinoma, wherein the C / C genotype is predicted not to respond to sorapenib as compared with the C / C genotype and the G / G genotype / RTI >

(SNP) of the 26th base of the sequence shown in SEQ ID NO: 2 in a gene sample obtained from a malignant neoplasm patient, and has a C / C genotype and a G / G genotype, The results of this study were as follows. First,
The present invention provides a method of treating a patient suffering from a malignant neoplasm that has a C / G genotype and is predicted not to respond to an anticancer chemotherapy system having the same mechanism as that of sorapenib, in comparison with a C / C genotype and a G / And to provide information for predicting the reactivity and survival prognosis of chemotherapy with the same mechanism.

13. The method according to claim 12, wherein the chemotherapy regimen having the same mechanism as that of sorapenib is regorafenib, Lenvatinib, Donafenib, or Brivanib. .

A composition for predicting the reactivity and survival prognosis of sorapenib in patients with hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof .

2 or a complementary polynucleotide consisting of 10 or more contiguous bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof as an essential component, For the prediction of reactivity and survival prognosis.

16. The composition according to claim 15, wherein the primer pair sequence comprises the nucleotide sequences of SEQ ID NOS: 5 to 6 or SEQ ID NOS: 11 to 12, and for predicting the reactivity and survival prognosis of sorapenib in patients with hepatocellular carcinoma.

[16] The composition according to claim 15, wherein the probe sequence comprises the nucleotide sequence of SEQ ID NO: 13 or SEQ ID NO: 14.

A kit for predicting the reactivity and survival prognosis of sorapenib in patients with hepatocellular carcinoma comprising a polynucleotide consisting of 10 or more consecutive bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof .

2 or a complementary polynucleotide consisting of 10 or more contiguous bases comprising the 26th base of the sequence represented by SEQ ID NO: 2 or a complementary polynucleotide thereof as an essential component, Kits for the prediction of reactivity and survival prognosis.

20. The method according to claim 19, wherein the primer pair sequence comprises the nucleotide sequence of SEQ ID NO: 5 to 6 or SEQ ID NO: 11 to 12, and the probe sequence comprises the nucleotide sequence of SEQ ID NO: Kit for prediction of reactivity and survival prognosis for sorafenib.