KR20170052454A - Biomarker composition for predicting sensitivity of sorafenib - Google Patents

Abstract

The present invention relates to a biomarker composition for predicting the sensitivity of sorafenib. More specifically, hepatocellular carcinoma with a high expression of fibrinogen-like protein 1 (FGL1) was verified to have high sensitivity to sorafenib, and thus the FGL1 can be applied and used as a biomarker composition for predicting sensitivity of sorafenib which predicts a therapeutic response to sorafenib. In addition, the present invention can provide a kit for predicting the sensitivity of sorafenib comprising an agent capable of detecting the level of expression or activity of the FGL1 as an active ingredient. Moreover, the present invention can be utilized as a pharmaceutical composition for preventing or treating liver cancer containing sorafenib and a promoter for the expression or activity of the FGL1. Also, provided is an information providing method for the prediction of sorafenib sensitivity comprising a step of measuring the expression or activity of the FGL1.

Description

Technical Field [0001] The present invention relates to a biomarker composition for predicting susceptibility to sorafenib,

The present invention relates to a biomarker composition for predicting the susapenib sensitivity, and more particularly, to a method for predicting the therapeutic response to sorafenib using the expression level of fibrinogen-like protein 1 (FGL 1) To a biomarker composition for prediction.

For the treatment of cancer, surgical therapy, radiation therapy and chemotherapy are used. Today, about 60 different types of anticancer drugs are being used. As knowledge of cancer development and cancer cell characteristics is well known, research on the development of new anticancer drugs is actively under way. However, when cancer drugs are administered repeatedly over a long period of time or when cancer is recurred, there is a disadvantage that cancer cells lose their therapeutic effect by acquiring resistance to anticancer drugs. In addition, most anticancer drugs have the effect of inhibiting the synthesis of nucleic acid in the cell or directly binding to nucleic acid to impair its function. These anticancer drugs do not only act selectively on cancer cells, but also on normal cells, Which causes various side effects such as a decrease in bone marrow function, gastrointestinal mucosal damage, hair loss, and the like.

Therefore, the above-mentioned anticancer drugs are required to develop various kinds of drugs due to their resistance properties, and in particular, there is a need for selective treatment of anticancer drugs that can minimize side effects of patients.

In order to overcome the disadvantages of chemotherapy, individual SNPs (single nucleotide polymorphism) have been analyzed and various attempts have been made to administer appropriate anticancer drugs accordingly. In addition, the development trend of new anticancer drugs is increasing in proportion to the development of targeted anticancer drugs, biopharmaceuticals, preventive vaccines and diagnostic agents. In order to increase the compliance of patients with cancer, the proportion of development of oral anticancer drugs is increasing.

Target cancer drugs can not kill cancer cells. Instead, it is a drug that inhibits the growth and growth of cancer cells by suppressing the necessary factors for cancer cells to grow. Therefore, even patients who are unable to completely remove the cancer can prolong the survival time by slowing the progression of the cancer through targeted anticancer drugs. In addition, theoretically, since there is no toxicity to normal cells, the painful side effects are small and the quality of life is expected to be superior to the conventional anticancer drugs.

Sorapenib is a receptor tyrosine kinase that is expected to over-express tumor cells or tumor vasculature, vascular endothelial growth factor receptor (VEGFR), platelet derived growth factor receptor (VEGFR) PDGFR) and multiple kinase inhibitors (RAF kinase) that simultaneously inhibit the serine / threonine kinase (RAF kinase) in the signaling pathway. It is an oral multikinase inhibitor that attacks cancer and is the first approved drug in the FDA.

Sorapenib has been shown to have a statistically significant survival rate of 2.8 months in cancer patients in a placebo-controlled triple-phase study centered on Europe and the Americas. In patients with advanced hepatocellular carcinoma, It is recommended. Although advanced hepatocellular carcinoma (HCC) is very difficult to treat, various therapies such as systemic chemotherapy have been tried, but no medication has been proven to improve the overall survival rate in a randomized controlled clinical trial.

In addition, Sorapenib is undergoing clinical trials for a variety of solid tumors, which are already being used as anticancer agents in renal cell carcinoma. Clinical studies on advanced hepatocellular carcinoma have been conducted recently, and the US Food and Drug Administration recently filed a treatment for hepatocellular carcinoma . Sorapenip has achieved revenue of $ 373 million in the first half of 2013 and is currently approved for treatment of liver cancer and kidney cancer. The US FDA recently approved Nexavar's thyroid cancer treatment.

However, there is no method for predicting the response of the patient and a method for confirming the response before the treatment is started, and studies on reliable biomarkers are lacking There is a problem in that it is very difficult to predict the reactivity of the subject to treatment with sorafenib and administer the appropriate drug. Furthermore, it is still possible to predict the response of a subject to sorafenib treatment, which can reduce patient inconvenience and reduce treatment costs by achieving optimal therapeutic effect by administering appropriate sorafenib to substantially multiple patients with liver cancer There is a problem that research on the method is insufficient.

To improve the therapeutic efficacy of sorafenib, concurrent therapy, resistance factor and mechanism studies, and reactive marker studies are continuing, but there is no clinical predictor of therapeutic efficacy of sorafenib. Therefore, the development of a biomarker capable of predicting susceptibility to sorafenib is urgently required in order to suppress the tolerance to sorafenib and improve the therapeutic efficiency.

Korean Registered Patent No. 10-1598122 (registered on Feb. 22, 2016)

It is an object of the present invention to provide a biomarker composition for the prediction of sorafenib sensitivity comprising fibrinogen-like protein 1 (FGL 1) or a gene encoding the same as an active ingredient.

Another object of the present invention is to provide a sorafenib sensitivity prediction kit comprising as an active ingredient a preparation capable of detecting the level of expression or activity of fibrinogen-like protein 1.

It is still another object of the present invention to provide a pharmaceutical composition for improving the sensitivity to sorafenib comprising an expression or activity promoting agent for fibrinogen-like protein 1 as an active ingredient.

It is still another object of the present invention to provide a pharmaceutical composition for preventing or treating liver cancer comprising fibrinogen-like protein 1 expression promoter or activity promoter and sorapenib.

It is still another object of the present invention to provide a method for providing information for sorapenib sensitivity prediction, which comprises measuring the expression or activity of fibrinogen-like protein 1.

In order to achieve the above object, the present invention provides a biomarker composition for the susapenib sensitivity prediction comprising fibrinogen-like protein 1 (FGL 1) or a gene encoding the same as an active ingredient.

In addition, the present invention provides a kit for predicting the susapenib sensitivity comprising an agent capable of detecting the expression or activity level of fibrinogen-like protein 1 as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for enhancing sorapenib sensitivity comprising an expression or activity promoting agent for fibrinogen-like protein 1 as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for preventing or treating liver cancer comprising fibrinogen-like protein 1 expression or activity promoter and sorapenib.

The present invention also provides a method for providing information for sorafenib sensitivity prediction, comprising the step of measuring the expression or activity of fibrinogen-like protein 1.

The expression of fibrinogen-like protein 1 in hepatocellular carcinoma was higher than that of hepatocellular carcinoma in which expression of fibrinogen-like protein 1 was low. In addition, it was confirmed that when the expression level of fibrinogen-like protein 1 is decreased by artificially decreasing the expression level of fibrinogen-like protein 1 in hepatocellular carcinoma, the sensitivity to sorapenib is decreased. Accordingly, the present invention can be applied to a biomarker composition for predicting the therapeutic reactivity of sorapenib with the fibrinogen-like protein 1 or a gene encoding the same.

In addition, the present invention can provide a sorafenib sensitivity prediction kit comprising as an active ingredient a preparation capable of detecting the expression or activity level of fibrinogen-like protein 1.

In addition, the present invention can provide a pharmaceutical composition for enhancing sorapenib sensitivity comprising an expression or activity promoting agent of fibrinogen-like protein 1 as an active ingredient.

In addition, the present invention can provide a pharmaceutical composition for preventing or treating liver cancer comprising fibrinogen-like protein 1 expression or activity promoter and sorapenib.

In addition, the present invention can provide a method for providing information for sorafenib sensitivity prediction, which comprises measuring the expression or activity of fibrinogen-like protein 1.

Figure 1 is a Western blot result comparing the expression levels of endogenous fibrinogen-like protein 1 in hepatocellular carcinoma cell lines.
FIG. 2A shows the survival rate of sorapenib-treated hepatocellular carcinoma cell line by using MTT assay. After the treatment with sorapenib, the survival rate of hepatocellular carcinoma by sorapenib was examined.
FIG. 2B shows the result of treatment with sorapenib at a concentration of 10 μM in the survival analysis of the hepatocellular carcinoma cell line of FIG. 2A.
Fig. 3 shows results of flow cytometry comparing the cytotoxic effect of sorapenib on hepatocellular carcinoma cell lines.
FIG. 4 shows Western blot results comparing cell proliferation and apoptosis-related protein expression levels according to HCC cell lines.
FIG. 5A is a graph showing the number of cell clusters as an experiment on the result of colony formation survival analysis of hepatocellular carcinoma cell line when sorapenib was not treated. FIG.
FIG. 5B is a graph showing the formation of cell clusters as a percentage in an experiment on the colony forming survival analysis of the hepatocellular carcinoma cell line by sorapenib.
FIG. 6 is a graph showing the formation of cell clusters as a percentage of the colonization formation survival analysis of sorafenib-induced hepatocellular carcinoma cell line after inhibiting the expression of fibrinogen-like protein 1 using siRNA.

As used herein, the term " fibrinogen-like protein 1 " is sometimes referred to as the term hepassocin or hepatocyte-derived fibrinogen related protein (HFREP1). Expression in lung, heart, small intestine, kidney, and other tissues other than liver has not been reported. Recently, expression in some adipocytes has been reported. Fibrinogen-like protein 1 is expressed specifically in liver hepatocytes and is secreted into a homodimer structure. In addition, it is expected to play an important role in regeneration after hepatic resection with high expression level during liver regeneration. Another report suggests that the expression level of fibrinogen-like protein 1 in malignant liver tumor cells is regulated to a low level and is likely to be associated with tumor cell differentiation.

The term " sorapenib " as used herein refers to a vascular endothelial growth factor receptor (VEGFR), a platelet derived growth factor receptor (PDGFR), a signaling pathway serine / threonine kinase / threonine kinase (RAF kinase), which is a multiple kinase inhibitor that simultaneously targets vascular endothelial cells that nourish cancer cells and cancer cells and treats cancer by administering an oral multikinase inhibitor ) And is the first drug approved by the FDA.

As used herein, the term " biomarker " refers to a substance capable of distinguishing a tissue or cell in which a cancer has developed from a normal cell or a tissue or cell subjected to appropriate cancer therapy, An organic biomolecule such as a protein or nucleic acid, a lipid, a glycolipid, a glycoprotein, or the like that exhibits an increase or a decrease.

As used herein, the term " hepatocellular carcinoma " generally refers to a cancer originating in hepatocytes. About 90% of malignant tumors in the liver are hepatocellular carcinoma, about 10% are cholangiocarcinoma, and a few others are other cancers. Hepatocellular carcinoma is the most common, so 'liver cancer' refers to hepatocellular carcinoma. Hepatocellular carcinoma (HCC) is a metastatic liver cancer that develops in the liver from primary liver cancer occurring in the liver and cancer occurring in other tissues. Most of the causes are unclear, but liver cirrhosis is often present, and hepatocellular carcinoma is found to occur in patients with cirrhosis, chronic active hepatitis B, or hepatitis B carriers. The present inventors confirmed that when the autoantibody marker of the present invention is used, diagnosis can be made with high sensitivity and specificity for the incidence of liver cancer from the individual.

As used herein, the term "susceptibility " refers to the rate at which a disease entity is positively detected, and when a diagnostic method is used, it is a criterion for how well the subject is actually found.

As used herein, the term " prophylactic " means any action that inhibits cancer or delays the onset of cancer by the pefrinocin-like protein 1 or a gene encoding the same.

As used herein, the term " treatment " refers to any action that improves or alleviates the symptoms of cancer by the fibrinogen-like protein 1 or a gene encoding the same.

The term " primer " as used herein means a short nucleic acid sequence capable of forming a base pair with a complementary template with a short free 3-terminal hydroxyl group and functioning as a starting point for template strand replication . The primer can initiate DNA synthesis in the presence of reagents and four different nucleoside triphosphates for polymerization reactions (i. E., DNA polymerase or reverse transcriptase) at appropriate buffer solutions and temperatures. In addition, primers may incorporate additional features that do not alter the primer properties of the primers that serve as a starting point for DNA synthesis, as sense and antisense nucleic acids with 7 to 50 nucleotide sequences.

As used herein, the term " probe " means a nucleic acid fragment such as RNA or DNA corresponding to a few nucleotides or hundreds of nucleotides that can specifically bind to an mRNA. , And the amount of expression can be confirmed. The probe may be prepared in the form of an oligonucleotide probe, a single strand DNA probe, a double strand DNA probe, or an RNA probe. Selection of suitable probes and hybridization conditions can be appropriately selected according to techniques known in the art.

The term "antibody" as used herein refers to a specific immunoglobulin directed against an antigenic site, wherein an antibody in the present invention refers to an antibody that specifically binds to fibrinogen-like protein 1, Any of those prepared by a conventional method or commercially available may be used. In addition, the antibody includes a polyclonal antibody, a monoclonal antibody, and a fragment capable of binding to an epitope. The antibody refers to a complete form having two full-length light chains and two long-length heavy chains, and also includes special antibodies such as humanized antibodies.

In addition, the kit of the present invention comprises an antibody that specifically binds to a marker component, a secondary antibody conjugate conjugated with a labeling substance that is colored by the reaction with the substrate, a coloring substrate solution that reacts with the labeling substance, Enzyme reaction termination solutions, and the like, and can be made from a number of separate packaging or compartments, including the reagent components used.

As used herein, the term "peptide" has a high binding capacity to the target material and does not cause denaturation during thermal / chemical treatment. Also, because of its small size, it can be used as a fusion protein by attaching it to other proteins. It can be used as a diagnostic kit and a drug delivery material because it can be specifically attached to a polymer protein chain.

As used herein, the term "aptamer" is a single-stranded nucleic acid (DNA, RNA or modified nucleic acid) having a stable tertiary structure and capable of binding to a target molecule with high affinity and specificity . Aptamers are comparable to monoclonal antibodies due to their inherent high affinity (usually pM levels) and their ability to bind to target molecules with specificity, and there is a high likelihood of being an alternative antibody, especially as a "chemoantibody".

The present invention provides a biomarker composition for the susapenib sensitivity prediction comprising fibrinogen-like protein 1 (FGL 1) or a gene encoding the same as an active ingredient.

Preferably, the fibrinogen-like protein 1 may be, but is not limited to, the amino acid sequence shown in SEQ ID NO: 1.

Preferably, the gene may be, but is not limited to, the nucleotide sequence shown in SEQ ID NO: 2.

Preferably, the expression or activity increase of the fibrinogen-like protein 1 may be susceptible to sorapenib.

Preferably, the sorapenib is a target anticancer agent for the treatment of cancer, wherein the cancer is selected from the group consisting of liver cancer, thyroid cancer, kidney cancer, esophageal cancer, gastric cancer, pancreatic cancer, small bowel cancer, colon cancer, rectal cancer, lung cancer, Cancer, and brain cancer. More specifically, the liver cancer may be, but is not limited to, hepatocellular carcinoma, biliary epithelial carcinoma, hepatoblastoma, and angiosarcoma.

Preferably, the expression or activity level of the fibrinogen-like protein 1 is determined by reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), radioimmunoassay but is not limited to, any one selected from the group consisting of radioimmunoassay, immunohistochemistry, microarray, western blotting and flow cytometry (FACS).

The present invention provides a sorafenib sensitivity prediction kit comprising as an active ingredient a preparation capable of detecting the expression or activity level of fibrinogen-like protein 1.

 Preferably, the agent may be selected from the group consisting of primers, probes, antibodies, peptides, and aptamers, but is not limited thereto.

The present invention provides a pharmaceutical composition for enhancing sorapenib sensitivity comprising an expression or activity promoting agent of fibrinogen-like protein 1 as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating liver cancer comprising fibrinogen-like protein 1 expression or activity promoter and sorapenib.

When the composition of the present invention is a pharmaceutical composition, it may contain a pharmaceutically acceptable carrier in addition to the above-mentioned active ingredients. Such pharmaceutically acceptable carriers are those conventionally used in pharmaceutical preparations, and include lactose, dextrose, The present invention relates to a process for the preparation of a medicament for the treatment and / or prophylaxis of cancer, comprising administering a therapeutically effective amount of a compound selected from the group consisting of cross, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, Carboxymethylcellulose, carboxymethylcellulose, hydroxybenzoate, talc, magnesium stearate, mineral oil, and the like. The pharmaceutical composition may further contain a filler, a weight agent, a binder, a lubricant, a wetting agent, a disintegrant, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, a fragrance, a preservative and the like as an additive and an auxiliary agent.

The pharmaceutical composition may be formulated into tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, nonaqueous solutions, emulsions, lyophilized preparations, suppositories and sterile injectable solutions. The pharmaceutical composition of the present invention may be administered orally or parenterally such as intravenously, intraarterally, intraperitoneally, intramuscularly, intrasternally, subcutaneously, intradermally, intranasally, intrapulmonary, rectally, topically, RTI ID = 0.0 >

The method of administration of the pharmaceutical composition is determined depending on the severity of symptoms, and local administration is generally preferred. The effective amount of the active ingredient of the above pharmaceutical composition means an amount required for prevention or treatment of the disease. Accordingly, the present invention is not limited to the particular type of the disease, the severity of the disease, the kind and amount of the active ingredient and other ingredients contained in the composition, the type of formulation and the patient's age, body weight, general health status, sex and diet, But is not limited to, the rate of administration, the rate of administration, the duration of the treatment, the drugs used concurrently, and the like.

The present invention provides a method for providing information for sorafenib sensitivity prediction, comprising the step of measuring the expression or activity of fibrinogen-like protein 1.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by 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  One : Hepatocellular carcinoma  Cell culture and Sorapanib  purchase

Human hepatocellular carcinoma cell lines (HepG2, Hep3B, Huh7, PLC / PRF / 5, SNU387, SNU449 and SNU475) were purchased from ACTT and Korean Cell Line Bank. and incubated in an incubator of% CO ₂ conditions.

Sorapenib was purchased from Selleckchem and purchased 10 mM sorapenib dissolved in 1 ml dimethyl sulfoxide (DMSO) and diluted in the medium immediately before the experiment.

Example  2 : Hepatocellular carcinoma  Depending on the cell line Intrinsic FGL  1 protein expression analysis

In order to analyze the expression level of the intracellular endogenous FGL 1 protein, sorafenib was treated with each cell line of hepatocellular carcinoma cells in a concentration-dependent manner, and the cells were lysed using a RIPA lysis solution, and the protein was quantified . A certain amount of protein was separated by molecular weight through PAGE (polyacrylamide gel electrophoresis) containing SDS (sodium dodecyl sulfate). Proteins were transferred to a nitrocellulose membrane and analyzed by immunoblotting.

As a result, referring to FIG. 1, it was confirmed that the expression levels of endogenous FGL 1 were different in seven hepatocellular carcinomas. Seven cell lines were selected from the cell lines (test; HepG2, Hep3B and Huh7), which express highly intrinsic FGL 1 protein, cell lines (gray; PLC / PRF / 5), and endogenous FGL 1 (White; SNU387, SNU449 and SNU475) in which the protein was almost not expressed.

Example  3: At Sorapenip  by Hepatocellular carcinoma  Cell line Growth rate proliferation analysis

In order to confirm the inhibitory activity of hepatocellular carcinoma proliferation by sorapenib, each cell line of hepatocellular carcinoma cells was plated in a 96-well plate, treated with sorafenib at 0, 2.5, 5, 10, 25 and 50 μM And cultured for 48 hours. Cells were treated with a solution of MTT (3- [4,5-dimethylthiazol-2yl] - 2,5-diphenyltetrazolium bromide) and reacted for 2 hours. Then, 50 μl of dimethylsulfoxide (DMSO) Absorbance was measured at a wavelength of 570 nm using a microplate reader (ELISA reader). Cell viability compared to normal control was expressed as percentage (%).

As a result, referring to FIG. 2, the cell lines (SNU387, SNU449 and SNU475) showing almost the expression of endogenous FGL 1 in the treatment with sorafenib 10 μM (HepG2, Hep3B, Huh7 and PLC / PRF / 5), and the cell survival rate of the cell line expressing endogenous FGL 1 was found to be decreased.

Example  4 : At Sorapenip  by Hepatocellular carcinoma  Analysis of cell killing effect

Each cell line of hepatocellular carcinoma was cultured in a 10 cm ² culture vessel followed by treatment with sorapenib at a concentration of 10 μM for 48 hours. Then, the cells were collected by treatment with trypsin / EDTA, centrifuged and the remaining cell pellet was transformed into a single cell. In order to stain the cell nuclei, PI (propidium iodide) was treated with 2 μg / ml and the number of killed cells was measured by flow cytometry. The cell death rate compared to the normal control was expressed as a percentage (%) .

Hep3, Hep3B, Huh7, and PLC / PRF (HepG2, Hep3B, Huh7, and Huh7) showing the expression of endogenous FGL 1 were observed in the cell lines (white; SNU387, SNU449 and SNU475) / 5) and 15% or more cell death was decreased compared with that of the control group. The cell line showing the expression of intrinsic FGL 1 showed increased cell death.

Example  5: Hepatocellular carcinoma  Analysis of cell proliferation and apoptosis related protein expression by cell line

In order to analyze the cell proliferation and apoptosis-related protein expression according to the hepatocellular carcinoma cell line, each cell line of hepatocellular carcinoma cell line was treated with 10 μM sorapanib and then the cells were dissolved using a RIPA lysis solution , And the protein was quantified. A certain amount of protein was separated by molecular weight through PAGE (polyacrylamide gel electrophoresis) containing SDS (sodium dodecyl sulfate). Proteins were transferred to a nitrocellulose membrane and analyzed by immunoblotting.

PCNA and p-ERK are involved in cell proliferation. LC3 (increased form II), cleaved PARP, and cleaved caspase 3 are proteins involved in apoptosis. Referring to FIG. 4, it was confirmed that sorafenib treatment inhibited phosphorylation of ERK and induced accumulation of autolytic cells in hepatocellular carcinoma cell lines (HepG2 and Huh7) with high expression of FGL1.

Example  6: At Sorapenip  by Hepatocellular carcinoma  Cell line Colony formation  Clonogenic survival assay

Each cell line of hepatocellular carcinoma was cultured in a 6 cm 2 incubator, treated with sorafenib at 0, 2, 5 μM, and cultured for 10 days. After 10 days, colonies were formed. The cells were washed with phosphate buffered saline (PBS), and fixed with methanol and acetone mixed with 2: 3 (volume ratio). Thereafter, the cells were stained with 0.4% crystal violet, and then washed twice with distilled water to confirm the number of cell clusters.

As a result, FIG. 5A shows the number of cell clusters formed on the 10th day after the same number of cells were placed in the culture container, and FIG. 5B shows the result of the formation of cell clusters generated after treatment with sorapenib As compared with the control group. In the cell lines (SNU387, SNU449 and SNU475) with little expression of FGL 1, the resistance to sorapenib was higher than that of the cell lines with high expression of FGL 1 (HepG2, Hep3B and Huh7).

Example  7: To siRNA  by FGL  1 expression, At Sorapenip  by Hepatocellular carcinoma  Cell line Colony formation  Survival analysis

The small interfering RNA of FGL 1 was purchased from IDT and the expression of FGL 1 was reduced by the injection of Hep3B cells with high expression of endogenous FGL 1 using lipofectamine 2000. Then, 0, 2 μM sorapenib was treated and colony formation survival analysis was compared.

As a result, referring to FIG. 6, FGL 1 siRNA increased the survival rate of Hep3B colonization and significantly increased the resistance to sorapenib. This confirms that the expression level of FGL 1 is closely related to the resistance of sorapenib.

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

The scope of the present invention is defined by the appended claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included within the scope of the present invention.

<110> KOREA INSTITUTE OF RADIOLOGICAL & MEDICAL SCIENCES <120> Biomarker composition for predicting sensitivity of sorafenib <130> ADP-2016-0062 <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 312 <212> PRT <213> fibrinogen like 1 <400> 1 Met Ala Lys Val Phe Ser Phe Ile Leu Val Thr Thr Ala Leu Thr Met   1 5 10 15 Gly Arg Glu Ile Ser Ala Leu Glu Asp Cys Ala Gln Glu Gln Met Arg              20 25 30 Leu Arg Ala Gln Val Arg Leu Leu Glu Thr Arg Val Lys Gln Gln Gln          35 40 45 Val Lys Ile Lys Gln Leu Leu Gln Glu Asn Glu Val Gln Phe Leu Asp      50 55 60 Lys Gly Asp Glu Asn Thr Val Ile Asp Leu Gly Ser Lys Arg Gln Tyr  65 70 75 80 Ala Asp Cys Ser Glu Ile Phe Asn Asp Gly Tyr Lys Leu Ser Gly Phe                  85 90 95 Tyr Lys Ile Lys Pro Leu Gln Ser Pro Ala Glu Phe Ser Val Tyr Cys             100 105 110 Asp Met Ser Asp Gly Gly Gly Trp Thr Val Ile Gln Arg Arg Ser Asp         115 120 125 Gly Ser Glu Asn Phe Asn Arg Gly Trp Lys Asp Tyr Glu Asn Gly Phe     130 135 140 Gly Asn Phe Val Gln Lys His Gly Glu Tyr Trp Leu Gly Asn Lys Asn 145 150 155 160 Leu His Phe Leu Thr Thr Gln Glu Asp Tyr Thr Leu Lys Ile Asp Leu                 165 170 175 Ala Asp Phe Glu Lys Asn Ser Arg Tyr Ala Gln Tyr Lys Asn Phe Lys             180 185 190 Val Gly Asp Glu Lys Asn Phe Tyr Glu Leu Asn Ile Gly Glu Tyr Ser         195 200 205 Gly Thr Ala Gly Asp Ser Leu Ala Gly Asn Phe His Pro Glu Val Gln     210 215 220 Trp Trp Ala Ser His Gln Arg Met Lys Phe Ser Thr Trp Asp Arg Asp 225 230 235 240 His Asp Asn Tyr Glu Gly Asn Cys Ala Glu Glu Asp Gln Ser Gly Trp                 245 250 255 Trp Phe Asn Arg Cys His Ser Ala Asn Leu Asn Gly Val Tyr Tyr Ser             260 265 270 Gly Pro Tyr Thr Ala Lys Thr Asp Asn Gly Ile Val Trp Tyr Thr Trp         275 280 285 His Gly Trp Trp Tyr Ser Leu Lys Ser Val Val Met Lys Ile Arg Pro     290 295 300 Asn Asp Phe Ile Pro Asn Val Ile 305 310 <210> 2 <211> 939 <212> DNA <213> fibrinogen like 1 <400> 2 atggcaaagg tgttcagttt catccttgtt accaccgctc tgacaatggg cagggaaatt 60 tcggcgctcg aggactgtgc ccaggagcag atgcggctca gagcccaggt gcgcctgctt 120 gagacccggg tcaaacagca acaggtcaag atcaagcagc ttttgcagga gaatgaagtc 180 cagttccttg ataaaggaga tgagaatact gtcattgatc ttggaagcaa gaggcagtat 240 gcagattgtt cagagatttt caatgatggg tataagctca gtggatttta caaaatcaaa 300 cctctccaga gcccagcaga attttctgtt tattgtgaca tgtccgatgg aggaggatgg 360 actgtaattc agagacgatc tgatggcagt gaaaacttta acagaggatg gaaagactat 420 gaaaatggct ttggaaattt tgtccaaaaa catggtgaat attggctggg caataaaaat 480 cttcacttct tgaccactca agaagactac actttaaaaa tcgaccttgc agattttgaa 540 aaaaatagcc gttatgcaca atataagaat ttcaaagttg gagatgaaaa gaatttctac 600 gagttgaata ttggggaata ttctggaaca gctggagatt cccttgcggg gaattttcat 660 cctgaggtgc agtggtgggc tagtcaccaa agaatgaaat tcagcacgtg ggacagagat 720 catgacaact atgaagggaa ctgcgcagaa gaagatcagt ctggctggtg gtttaacagg 780 tgtcactctg caaacctgaa tggtgtatac tacagcggcc cctacacggc taaaacagac 840 aatgggattg tctggtacac ctggcatggg tggtggtatt ctctgaaatc tgtggttatg 900 aaaattaggc caaatgattt tattccaaat gtaatttaa 939

Claims (13)

A biomarker composition for the prediction of susapenib susceptibility comprising fibrinogen-like protein 1 (FGL 1) or a gene encoding the same as an active ingredient. 2. The biomarker composition according to claim 1, wherein the fibrinogen-like protein 1 has the amino acid sequence of SEQ ID NO: 1. [Claim 2] The biomarker composition of claim 1, wherein the gene has the nucleotide sequence of SEQ ID NO: 2. The biomarker composition of claim 1, wherein the expression or activity increase of the fibrinogen-like protein 1 is susceptible to sorapenib. 2. The biomarker composition of claim 1, wherein the sorapenib is a target anticancer drug for cancer therapy. The method of claim 5, wherein the cancer is selected from the group consisting of liver cancer, thyroid cancer, kidney cancer, esophageal cancer, gastric cancer, pancreatic cancer, small bowel cancer, colon cancer, rectal cancer, lung cancer, breast cancer, uterine cancer, ovarian cancer, prostate cancer, Wherein the biomarker composition is for use in predicting susceptibility to sorafenib. The biomarker composition according to claim 6, wherein the liver cancer is selected from the group consisting of hepatocellular carcinoma, biliary epithelial carcinoma, hepatoblastoma, and angiosarcoma. The method according to claim 1, wherein the expression or activity level of the fibrinogen-like protein 1 is determined by reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) Characterized in that the measurement is carried out by any one selected from the group consisting of radioimmunoassay, immunohistochemistry, microarray, western blotting and flow cytometry (FACS). Composition. A kit for predicting susapepub sensitivity comprising an agent capable of detecting the expression or activity level of fibrinogen-like protein 1 as an active ingredient. The kit according to claim 9, wherein the agent is selected from the group consisting of primers, probes, antibodies, peptides, and aptamers. A pharmaceutical composition for enhancing susapepub sensitivity comprising an expression or activity promoter of fibrinogen-like protein 1 as an active ingredient. A pharmaceutical composition for the prevention or treatment of liver cancer comprising the expression or activity promoter of fibrinogen-like protein 1 and sorapenib. And measuring the expression or activity of the fibrinogen-like protein 1.

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