KR102395580B1 - Companion diagnostic biomarker composition and companion diagnostic kit comprising the same - Google Patents

Abstract

The present invention relates to a biomarker composition for companion diagnosis and a kit for companion diagnosis comprising the same, and more specifically, one of PD-1 checkpoint inhibitor (PD-1 Checkpoint Inhibitor) and PD-L1 immune checkpoint inhibitor (PD-L1 Checkpoint Inhibitor). It relates to a companion diagnostic biomarker composition for predicting a therapeutic response to any one or more immune checkpoint inhibitors, and a companion diagnostic kit comprising the same. According to the present invention, it is possible to predict the therapeutic response of any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor through proteomic analysis in the blood of a cancer patient as well as a companion diagnosis through the tissue of a cancer patient. can have an effect. In addition, the biomarker composition for companion diagnosis and the companion diagnosis kit comprising the same according to the present invention include a reactive patient group and It has the advantage of high accuracy in screening non-responder patients who do not show the effect.

Description

A biomarker composition for companion diagnosis and a kit for companion diagnosis comprising the same

The present invention relates to a biomarker composition for companion diagnosis and a kit for companion diagnosis comprising the same, and more specifically, one of PD-1 checkpoint inhibitor (PD-1 Checkpoint Inhibitor) and PD-L1 immune checkpoint inhibitor (PD-L1 Checkpoint Inhibitor). It relates to a companion diagnostic biomarker composition for predicting a therapeutic response to any one or more immune checkpoint inhibitors, and a companion diagnostic kit comprising the same.

Companion Diagnosis is a diagnostic technique for predicting a patient's responsiveness to a specific drug treatment in advance. Targeted anticancer drugs that selectively attack specific target proteins in order to overcome the shortcomings of most existing anticancer drugs, which have large side effects by acting on both cancer cells and normal cells, and immune anticancer drugs that kill cancer cells by activating the suppressed immune system etc have been developed.

However, since targeted anticancer drugs are only effective for cancer patients having a specific target protein even for the same type of cancer, treatment efficiency is very low if patients with such target protein are not selected. In addition, since targeted anticancer drugs depend on inhibition of cell growth and proliferation rather than apoptosis, the possibility of resistance development due to continuous drug administration over a long period of time is high. Therefore, it is necessary to select a group of patients showing an effect on the drug before administering the drug through the analysis of the target of the anticancer drug.

Immunocancer drugs exhibit anticancer effects by enhancing specificity, memory, and adaptiveness of the immune system. In other words, it uses the body's immune system to precisely attack cancer cells, so there are fewer side effects, and because the immune system's memory and adaptability are used, patients who are effective against immune anticancer drugs can see continuous anticancer effects. However, even with the same type of cancer, the degree of anticancer effect differs depending on the specific patient, so a companion diagnosis is required to predict the patient's therapeutic response through the immunotherapy.

A companion diagnostic biomarker is used to select a patient group that is effective in a specific drug treatment. Here, a companion diagnostic biomarker is an index for predicting the patient's reactivity to a specific drug treatment in advance. Protein, DNA, RNA, metabolites, etc. may include That is, in the case of a specific disease or cancer, it refers to a marker that can distinguish between normal and pathological conditions, predict treatment response, and objectively measure.

Meanwhile, as pharmaceutical companies reduce the cost of developing new drugs and increase the demand for targeted anticancer drugs, the global market for companion diagnostics is expected to grow at an annual rate of 18% between 2013 and 2019, reaching approximately $5.8 billion in 2019. It is estimated

For example, Roche, one of the multinational pharmaceutical companies, acquired Genentech, which developed "Herceptin," the first breast cancer-targeted anticancer drug, and "Herceptest," a companion diagnostic kit for it, to start companion diagnosis-based anticancer drug treatment.

The companion diagnostic kit is a method to check the overexpression of a specific protein through immunohistochemical tests such as DAKO and HercepTest, and FISH or CISH test using a DNA probe for gene amplification of a specific gene such as Ventana Medical Systems, INFORM HER-2/NEU There is a method for confirming the biomarker gene by using genomic techniques such as q-PCR such as Roche Diagnostics and cobas EGFR mutation test to check whether the biomarker gene is mutated or not.

On the other hand, immunotherapy can be divided into passive immunotherapy and active immunotherapy. Passive immunotherapy includes an immune checkpoint inhibitor, immune cell therapy, and therapeutic antibody. Immune checkpoint inhibitors are drugs that attack cancer cells by activating T cells by blocking the activation of immune checkpoint proteins involved in T cell suppression, and include CTLA-4, PD-1, and PD-L1 immune checkpoint inhibitors. .

PD-1 immune checkpoint inhibitors are second-generation immune cancer drugs, Pembrolizumab (KEYTRUDA ^® ) developed by Merck and Nivolumab (OPDIVO ^® ) developed by Ono Pharmaceutical and BMS. Atezolizumab (Tecentriq ^® ), a PD-L1 immune checkpoint inhibitor developed by Genentech/Roche, was approved by the US FDA in 2014.

Immunity with human anti-PD-1 monoclonal antibody or human anti-PD-L1 monoclonal antibody that blocks PD-1 mediated lymphocyte negative regulation (interaction between PD-1 and PD-L1 and PD-L2 ligand) It has a mechanism of action that recognizes cancer cells as foreign substances and removes them.

Pembrolizumab is used in Malignant Melanoma, Non-Small Cell Lung Cancer, Head And Neck Squamous Cell Cancer, classic Hodgkin Lymphoma, urothelial cancer ( FDA-approved for the treatment of Urothelial Carcinoma, Deficient Mismatch Repair (dMMR) or any solid cancer with high frequency Microsatellite Instability-High (MSI-High), and adenocarcinoma of the stomach and gastroesophageal junction (Adenocarcinoma) Nivolumab has shown clinical benefit in clinical studies for various advanced cancers, and has recently been approved as a drug in melanoma, non-small cell lung cancer, and kidney cancer, and is widely used.

Furthermore, the “PD-L1 IHC 22C3 pharmDx test” is a companion diagnostic test used to determine whether pembrolizumab effectively exhibits anticancer effects as an immunotherapy for specific patients. There is this. The PD-L1 IHC 22C3 pharmDx test is one of the immunohistochemistry (IHC) methods. Using DAKO's PD-L1 IHC 22C3 pharmDx product, the PD-L1 expression rate of biopsied cancer cells was observed to be suitable for treatment of pembrolizumab. It is a test method used to select a group of patients. Similarly, there are “VENTANA PD-L1 (SP263) Assay test” and “PD-L1 IHC 28-8 pharmDx test” as companion diagnostic tests for nivolumab.

However, these existing companion diagnostic tests measure the expression of PD-L1 by removing some tissue (405 μm) from the patient's tumor. Not only does it take more time, but there is also a problem in that the accuracy of selecting a patient group showing an effect on the immune anticancer agent is somewhat inferior.

Accordingly, in order to overcome the technical limitations of the companion diagnostic test, the present inventors conducted PD-1 checkpoint inhibitor (PD-1 Checkpoint Inhibitor) and PD-L1 immune checkpoint inhibitor (PD) through measurement in tissue as well as proteomic analysis in blood. -L1 Checkpoint Inhibitor), a companion diagnostic biomarker composition for predicting the therapeutic response of any one or more immune checkpoint inhibitors and a companion diagnostic kit including the same were developed.

In order to solve the above problems, the present invention provides a PD-1 checkpoint inhibitor and PD-L1 through proteomic analysis in the blood of a cancer patient as well as a companion diagnosis through the tissue of a cancer patient. An object of the present invention is to provide a companion diagnostic biomarker composition for predicting a therapeutic response of any one or more immune checkpoint inhibitors among immune checkpoint inhibitors (PD-L1 Checkpoint Inhibitor) and a companion diagnostic kit comprising the same.

In addition, the present invention relates to a reactive patient group showing an effect on anticancer treatment through any one or more immune checkpoint inhibitors of a PD-1 checkpoint inhibitor (PD-1 Checkpoint Inhibitor) and a PD-L1 immune checkpoint inhibitor (PD-L1 Checkpoint Inhibitor) A biomarker composition for companion diagnosis for predicting the therapeutic response of any one or more immune checkpoint inhibitors among PD-1 immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors with high accuracy for screening non-responding patient groups that do not show the effect, and companion diagnostics composition comprising the same It is a second object to provide a kit for diagnosis.

Furthermore, the present invention provides a companion diagnostic biomarker composition capable of more accurately discriminating a patient group suitable for anticancer treatment through any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor, and companion diagnostics including the same The third purpose is to provide a kit.

In order to achieve the above object, the companion diagnostic biomarker composition according to the present invention includes complement C7 (Complement Component C7), and PD-1 (Programmed Cell Death Protein 1) immune checkpoint inhibitor and PD-L1 ( Programmed Death-Ligand 1) It is characterized in predicting the reactivity of any one or more immune checkpoint inhibitors among immune checkpoint inhibitors.

Here, the biomarker composition for companion diagnosis is HRG (Histidine-Rich Glycoprotein), AZGP1 (Zinc-Alpha-2-Glycoprotein), complement C5 (Complement Component C5), IHKV4-1 (Immunoglobulin Kappa Variable 4-1), MAN1A ( Mannosyl-Oligosaccharide 1,2-Alpha-Mannosidase 1A), A2M (Alpha-2-Macroglobulin), SPP1 (Osteopontin), and HYOU1 (Hypoxia Up-Regulated Protein 1) further comprising any one or more biomarkers selected from the group consisting of can do.

Here, the amount of the complement C7 protein may be measured through quantitative analysis.

Here, the protein amount of complement C7 is determined by protein mass spectrometry, protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization). Time of Flight Mass Spectrometry analysis, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry (Liquid) Chromatography-Mass Spectrometry, LC-MS), LC-MS/MS (Liquid Chromatography-Mass Spectrometry/ Mass Spectrometry), Western blot, and ELISA (Enzyme Linked Immunosorbentassay) can be measured by any one method selected from the group consisting of .

Here, the amount of complement C7 protein in the reactive patient group showing an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor can decrease.

Here, the amount of complement C7 protein in the non-responding patient group that does not show an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the reactive patient group showing the effect on anticancer treatment through the PD-1 immune checkpoint inhibitor can increase

Here, the amount of complement C7 protein in the reactive patient group showing an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor can decrease.

Here, the amount of complement C7 protein in the non-responsive patient group that does not show an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor compared to the reactive patient group showing the effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor can increase

Here, the cutoff value for the protein amount of complement C7 may be within the range of 100 to 110 μg/mL.

Here, when the amount of the complement C7 protein is equal to or greater than the cutoff value, the PD-1 immune checkpoint inhibitor may have low reactivity to the cancer cells.

Here, when the amount of the complement C7 protein is less than the cutoff value, the PD-1 immune checkpoint inhibitor may have high reactivity to the cancer cells.

Here, when the amount of the complement C7 protein is equal to or greater than the cutoff value, the PD-L1 immune checkpoint inhibitor may have low reactivity to the cancer cells.

Here, when the amount of the complement C7 protein is less than the cutoff value, the PD-L1 immune checkpoint inhibitor may have high reactivity to the cancer cells.

Here, the biomarker composition for companion diagnosis may be extracted from any one selected from the group consisting of blood, serum, plasma and tissue.

Here, the biomarker composition for companion diagnosis may be extracted from blood.

Here, the biomarker composition for companion diagnosis may be extracted from tissue.

Here, the biomarker composition for companion diagnosis may further include a Programmed Death-Ligand 1 (PD-L1) protein.

Here, the cancer cell may be a cancer cell corresponding to a carcinoma that specifically responds to one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor.

Here, the cancer cells are lung cancer, liver cancer, stomach cancer, gastric and gastroesophageal junction adenocarcinoma, skin melanoma, head and neck cancer, bone cancer, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, rectal cancer, colorectal cancer, colon cancer, breast cancer, uterine sarcoma, fallopian tube carcinoma, Endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, larynx cancer, small intestine cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, solid tumors of childhood, It may be a cancer cell corresponding to any one carcinoma selected from the group consisting of differentiated lymphoma, bladder cancer, renal cancer, renal cell carcinoma, renal pelvic carcinoma, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, and pituitary adenoma.

Here, the cancer cells may be cancer cells corresponding to lung cancer or liver cancer.

Here, the companion diagnostic biomarker composition may be administered concurrently or sequentially with any one or more immune checkpoint inhibitors of the PD-1 immune checkpoint inhibitor and the PD-L1 immune checkpoint inhibitor.

Meanwhile, the kit for companion diagnosis according to the present invention may include the biomarker composition for companion diagnosis according to the present invention.

The biomarker composition for companion diagnosis of the present invention and the companion diagnosis kit comprising the same according to the present invention as described above are not only a companion diagnosis through the tissue of a cancer patient, but also a PD-1 immune checkpoint inhibitor and PD-L1 through proteomic analysis in the blood of a cancer patient. There is an effect of predicting the therapeutic response of any one or more immune checkpoint inhibitors among the immune checkpoint inhibitors.

In addition, the biomarker composition for companion diagnosis and the companion diagnosis kit comprising the same according to the present invention include a reactive patient group and It has the advantage of high accuracy in screening non-responder patients who do not show the effect.

Furthermore, the biomarker composition for companion diagnosis according to the present invention and the companion diagnosis kit comprising the same more accurately determine a patient group suitable for anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor. There is an effect that can be done.

1 is a graph showing the results of a companion diagnosis predicting the reactivity of any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor against 9 types of companion diagnostic biomarkers including complement C7. it will be shown
2 is a graph showing the results of a companion diagnostic test using the biomarker composition for companion diagnostics according to an embodiment of the present invention.
3 is a graph showing a companion diagnostic test result when the biomarker IGKV4-1 is further included in the companion diagnostic biomarker composition according to an embodiment of the present invention.

The terms used in this application are only used to describe specific examples. Therefore, for example, a singular expression includes a plural expression unless the context clearly requires it to be singular. In addition, terms such as “include” or “include” as used in the present application are used to clearly indicate that the features, steps, functions, components, or combinations thereof described in the specification exist, and other features It should be noted that it is not intended to preliminarily exclude the existence of elements, steps, functions, components, or combinations thereof.

On the other hand, unless otherwise defined, all terms used herein should be regarded as having the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. Accordingly, unless explicitly defined herein, specific terms should not be construed in an unduly idealistic or formal sense.

As used herein, the term “companion diagnosis” refers to a diagnostic technique for predicting a patient's responsiveness to a specific drug treatment in advance.

As used herein, the term "biomarker for companion diagnosis" is an index for predicting the patient's reactivity to a specific drug treatment, and may include proteins, DNA, RNA, metabolites, and the like. That is, in the case of a specific disease or cancer, it refers to a marker that can distinguish between normal and pathological conditions, predict treatment response, and objectively measure.

As used herein, the term “immune checkpoint inhibitor” is synonymous with immune checkpoint inhibitor, which blocks the activation of immune checkpoint protein involved in T cell suppression to activate T cells to attack cancer cells. As a pharmaceutical, in the present specification, any one or more immune checkpoint inhibitors of a PD-1 checkpoint inhibitor and a PD-L1 checkpoint inhibitor are disclosed, and the PD-1 immune checkpoint inhibitor is disclosed. The inhibitor or PD-L1 immune checkpoint inhibitor may be, but is not limited to, pembrolizumab, nivolumabyl or atezolizumab.

<Biomarker composition for companion diagnosis>

The present inventors have devised the following invention as a result of research in order to solve the above-mentioned problem. The present specification includes complement C7 (Complement Component C7), and any one or more immune checkpoints of PD-1 Checkpoint Inhibitor and PD-L1 Checkpoint Inhibitor against cancer cells Disclosed is a biomarker composition for companion diagnosis, characterized in predicting the reactivity of an inhibitor.

The immune function in the body regulates the overall T cell function through the control of these co-stimulatory (Co-Stimulatory) and co-inhibitory (Co-Inhibitory) signals at the same time as antigen recognition. Therefore, immune cells detect tumor-specific antigens expressed due to changes such as mutations occurring in cancer cells and eliminate cancer cells. However, cancer cells try to suppress immune function by changing the tumor microenvironment to evade immune attack, or to evade immunity through T cell immune tolerance or immune editing. As one of these evasion strategies, it suppresses the function of T cells by changing the function of the immune checkpoint, that is, the immune checkpoint is a protein that prevents the destruction of cancer cells.

In the immune response of these T cells, the interaction of costimulatory receptors acting as accelerators, co-inhibiting receptors acting as brakes, and ligands binding to each receptor, operates very precisely in space and time. On the other hand, PD-1 of T cells regulates T cell function in peripheral tissues through PD-L1/PD-L2 of cancer cells. That is, when PD-L1 of cancer cells and PD-1 of T cells combine, T cells lose their function and die. Immune checkpoint inhibitors block the immune evasion signal by binding to the binding site of cancer cells and T cells, unlike existing immunotherapeutic agents such as cytokine therapeutics and anticancer vaccines. T cells have a mechanism to destroy cancer cells. That is, when the PD-1 antibody or the PD-L1 antibody binds in advance between the binding site of PD-L1 and PD-1, the immune evasion signal is blocked, and T cells that have not received the immune evasion signal kill cancer cells.

The companion diagnostic biomarker composition according to the present invention includes complement C7 (Complement Component C7), wherein C7 is a PD-1 checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor against cancer cells (PD-L1). L1 Checkpoint Inhibitor) can be used as a companion diagnostic biomarker predicting the reactivity of any one or more immune checkpoint inhibitors.

Complement component C7 (Complement component C7) included in the companion diagnostic biomarker composition according to the present invention serves as a biomarker to regulate the body's antigen-antibody immune response, and forms a membrane attack complex (MAC). dissolve pathogens; According to the classification of gene ontology, it is a protein involved in immune response and lysis, and its genetic information can be found in GeneBank Accession No., Uniprot, and the like. However, direct correlation with the prediction of complement C7 reactivity with PD-1 checkpoint inhibitor (PD-1 Checkpoint Inhibitor) or PD-L1 immune checkpoint inhibitor (PD-L1 Checkpoint Inhibitor) against cancer cells has not been disclosed anywhere.

In addition, the biomarker composition for companion diagnosis according to the present invention may further include an agent for measuring the amount of the complement C7 protein.

On the other hand, the biomarker composition for companion diagnosis according to the present invention is HRG (Histidine-Rich Glycoprotein), AZGP1 (Zinc-Alpha-2-Glycoprotein), complement C5 (Complement Component C5), IHKV4-1 (Immunoglobulin Kappa Variable 4-1) , MAN1A (Mannosyl-Oligosaccharide 1,2-Alpha-Mannosidase 1A), A2M (Alpha-2-Macroglobulin), SPP1 (Osteopontin) and HYOU1 (Hypoxia Up-Regulated Protein 1) any one or more biomarkers selected from the group consisting of may further include.

HRG (Histidine-Rich Glycoprotein), AZGP1 (Zinc-Alpha-2-Glycoprotein), complement C5 (Complement Component C5), IHKV4-1 (Immunoglobulin Kappa Variable 4-1) included in the companion diagnostic biomarker composition according to the present invention , MAN1A (Mannosyl-Oligosaccharide 1,2-Alpha-Mannosidase 1A), A2M (Alpha-2-Macroglobulin), SPP1 (Osteopontin) and HYOU1 (Hypoxia Up-Regulated Protein 1) are biomarkers for the human antigen-antibody immune response As a protein involved in immune response and lysis according to gene ontology classification, its genetic information can be found in GeneBank Accession No., Uniprot, etc. However, direct correlation with the prediction of the reactivity of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 against cancer cells of a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor has not been disclosed anywhere.

In addition, the biomarker composition for companion diagnosis according to the present invention further comprises an agent for measuring the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 can do.

Meanwhile, in the companion diagnostic biomarker composition according to the present invention, the amount of the complement C7 protein can be measured through quantitative analysis. Furthermore, the amount of complement C7 protein may be qualitatively analyzed, quantitatively analyzed, or both qualitative and quantitative analysis may be performed through proteomic analysis.

Similarly, in the biomarker composition for companion diagnosis according to the present invention, the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 can be measured through quantitative analysis. there is. Furthermore, the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M, and HYOU1 is quantitatively analyzed together with qualitative analysis, quantitative analysis and qualitative analysis through proteomic analysis. can also be done

As used herein, the term "protein amount" refers to a PD-1 immune checkpoint inhibitor and a PD- It refers to a process of confirming the presence and measurement level of proteins in the blood of the cancer patient by treating one or more immune checkpoint inhibitors among L1 immune checkpoint inhibitors. By using an antibody, interacting protein, ligand, nanoparticle or aptamer that specifically binds to the protein or peptide fragment, the amount of the protein can be confirmed as well as specific to the protein or peptide fragment It may include any detection means having a negative affinity (Affinity). More preferably, the amount of protein itself can be measured without using an antibody, an interacting protein, a ligand, nanoparticles, or an aptamer.

In the biomarker composition for companion diagnosis according to the present invention, the protein amount measurement or comparative analysis method includes protein mass spectrometry, protein chip analysis, immunoassay, ligand binding assay, and MALDI-TOF (Matrix Desorption/Ionization Time of Flight Mass). Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, octeroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, By two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry (LC-MS), LC-MS/MS (Liquid Chromatography-Mass Spectrometry/ Mass Spectrometry), Western blot and ELISA (Enzyme Linked Immunosorbentassay) It is preferable to measure or compare analysis by any one method selected from the group consisting of, but is not limited thereto.

On the other hand, the agent for measuring the amount of complement C7 protein in the companion diagnostic biomarker composition according to the present invention is from the group consisting of an antibody that specifically binds to the complement C7 protein, an interacting protein, a ligand, nanoparticles, and an aptamer. It may include any one or more selected. The antibody refers to a specific protein molecule directed against an antigenic site. For the purposes of the present invention, the antibody refers to an antibody that specifically binds to the complement C7 protein, polyclonal antibody, monoclonal antibody, and recombinant including all antibodies. The antibody can be easily prepared using techniques well known in the art. The antibody also includes functional fragments of the antibody molecule, as well as a complete form having two full-length light chains and two full-length heavy chains. The functional fragment of the antibody molecule refers to a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab') 2 and Fv.

In addition, in the biomarker composition for companion diagnosis according to the present invention, the agent for measuring the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is the HRG , AZGP1, complement C5, IHKV4-1, MAN1A, from the group consisting of an antibody that specifically binds to a protein of any one or more biomarkers selected from the group consisting of HYOU1, interacting proteins, ligands, nanoparticles and aptamers from the group consisting of It may include any one or more selected.

As used herein, "Aptamer" is a biopolymer material that inhibits protein interaction through three-dimensional binding with a specific target protein in the form of single or double-stranded DNA or RNA, which binds to various target molecules. have a characteristic Typically, an aptamer may be a small nucleic acid of 15-50 bases in length that folds into a defined secondary and tertiary structure, such as a stem-loop structure. Preferably, the aptamer binds to the target high- or low-expression protein with a kd of less than 10 ^-6 , 10 ^-8 , 10 ^-10 , or 10 ^-12 . The aptamer can bind to a high-expression protein or a low-expression protein having a very high degree of specificity, and the aptamer can be composed of a plurality of ribonucleotide units, deoxyribonucleotide units, or a mixture of two types of nucleotide residues. there is. In addition, the aptamer may further comprise one or more modified base, sugar or phosphate backbone units.

On the other hand, complement C7 included in the biomarker composition for companion diagnosis according to the present invention is effective in anticancer treatment through PD-1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through PD-1 immune checkpoint inhibitor It is characterized by a decrease in the amount of protein in the responding patient group showing

Conversely, complement C7 included in the companion diagnostic biomarker composition according to the present invention has no effect on anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the reactive patient group that shows the effect on anticancer treatment through the PD-1 immune checkpoint inhibitor. It is characterized by an increase in the amount of protein in the non-responding patient group.

In addition, complement C7 contained in the companion diagnostic biomarker composition according to the present invention is effective in anticancer treatment through PD-L1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through PD-L1 immune checkpoint inhibitor It is characterized by a decrease in the amount of protein in the responding patient group showing

Conversely, complement C7 contained in the companion diagnostic biomarker composition according to the present invention has no effect on anticancer treatment through PD-L1 immune checkpoint inhibitor compared to the reactive patient group that shows effect on anticancer treatment through PD-L1 immune checkpoint inhibitor. It is characterized by an increase in the amount of protein in the non-responding patient group.

In the biomarker composition for companion diagnosis according to the present invention, the cut-off value for the protein amount of complement C7 is preferably within the range of 90 to 120 μg/mL, and more specifically, within the range of 100 to 110 μg/mL. It is more preferable, but is not limited thereto. The cut-off value may be arbitrarily set by the performer performing the companion diagnostic test according to the number of cancer patients to be measured. The cutoff value for the protein amount of complement C7 is a standard value for judging the reactivity of any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor against cancer cells, and more specifically, PD- 1 Refers to a standard value for classifying into a reactive patient group that exhibits an effect on anticancer treatment through any one or more immune checkpoint inhibitors of an immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor and a non-responder group that does not show the effect.

Here, when the amount of the complement C7 protein is equal to or greater than the cutoff value, the PD-1 immune checkpoint inhibitor may have low reactivity to the cancer cells. That is, as a result of performing a companion diagnosis for predicting the reactivity of a PD-1 immune checkpoint inhibitor to cancer cells through the biomarker composition for companion diagnosis according to the present invention, when the amount of the complement C7 protein is greater than or equal to the cutoff value, the The reactivity of PD-1 immune checkpoint inhibitors is low. In this case, it can be classified as a non-responsive patient group that does not show an effect on chemotherapy through a PD-1 immune checkpoint inhibitor.

Conversely, when the amount of the complement C7 protein is less than the cutoff value, the PD-1 immune checkpoint inhibitor may have high reactivity against the cancer cells. That is, as a result of performing a companion diagnosis for predicting the reactivity of a PD-1 immune checkpoint inhibitor to cancer cells through the biomarker composition for companion diagnosis according to the present invention, when the amount of the complement C7 protein is less than the cut-off value, the The reactivity of PD-1 immune checkpoint inhibitors is high. In this case, it can be classified as a reactive patient group that is effective for anticancer treatment through PD-1 immune checkpoint inhibitor.

When the amount of the complement C7 protein is equal to or greater than the cutoff value, the PD-L1 immune checkpoint inhibitor may have low reactivity to the cancer cells. That is, as a result of performing a companion diagnosis for predicting the reactivity of a PD-L1 immune checkpoint inhibitor to cancer cells through the companion diagnostic biomarker composition according to the present invention, when the amount of the complement C7 protein is equal to or greater than the cutoff value, the This means that the reactivity of PD-L1 immune checkpoint inhibitors is low. In this case, it can be classified as a non-responder group that does not show an effect on chemotherapy through PD-L1 immune checkpoint inhibitors.

Conversely, when the amount of the complement C7 protein is less than the cutoff value, the PD-L1 immune checkpoint inhibitor may have high reactivity against the cancer cells. That is, as a result of performing a companion diagnosis for predicting the reactivity of a PD-L1 immune checkpoint inhibitor to cancer cells through the companion diagnostic biomarker composition according to the present invention, when the amount of the complement C7 protein is less than the cutoff value, the It means that the reactivity of the PD-L1 immune checkpoint inhibitor is high. In this case, it can be classified as a reactive patient group that is effective in anticancer treatment through PD-L1 immune checkpoint inhibitors. In this regard, specific details will be described in detail in the following {Examples and Evaluation}.

On the other hand, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 included in the companion diagnostic biomarker composition according to the present invention is a PD-1 immune checkpoint inhibitor and / Alternatively, compared to the non-responding patient group that does not show an effect on chemotherapy through the PD-L1 immune checkpoint inhibitor, the protein in the responding patient group showing an effect on chemotherapy through the PD-1 immune checkpoint inhibitor and/or the PD-L1 immune checkpoint inhibitor characterized in that the amount decreases.

Conversely, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 included in the companion diagnostic biomarker composition according to the present invention is a PD-1 immune checkpoint inhibitor and / Or, in the non-responding patient group that does not show an effect on chemotherapy through the PD-1 immune checkpoint inhibitor and/or PD-L1 immune checkpoint inhibitor, compared to the reactive patient group that shows an effect on chemotherapy through the PD-L1 immune checkpoint inhibitor. characterized in that the amount increases.

In addition, in the biomarker composition for companion diagnosis according to the present invention, the cutoff value for the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is It is preferably within the range of 50 to 150 ug/mL, but is not limited thereto. The cutoff value for the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M, and HYOU1 is a PD-1 immune checkpoint inhibitor and PD-L1 immunity against cancer cells. A value as a criterion for judging the reactivity of any one or more immune checkpoint inhibitors among the checkpoint inhibitors It means the standard value for classifying the responder group that shows and the non-responder group that does not show the effect.

Here, when the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is greater than or equal to the cutoff value, the PD-1 immune checkpoint inhibitor against the cancer cells and/or the PD-L1 immune checkpoint inhibitor may have low reactivity. That is, as a result of performing a companion diagnosis to predict the reactivity of a PD-1 immune checkpoint inhibitor and/or a PD-L1 immune checkpoint inhibitor to cancer cells through the companion diagnostic biomarker composition according to the present invention, the HRG, AZGP1, complement C5, When the protein amount of any one or more biomarkers selected from the group consisting of IHKV4-1, MAN1A, A2M and HYOU1 is greater than or equal to the cutoff value, the PD-1 immune checkpoint inhibitor and/or PD-L1 immune checkpoint inhibitor Reactivity means low. In this case, it can be classified into a non-responding patient group that does not show an effect on chemotherapy through a PD-1 immune checkpoint inhibitor and/or a PD-L1 immune checkpoint inhibitor.

Conversely, when the protein amount of any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is less than the cutoff value, the PD-1 immune checkpoint inhibitor against the cancer cell and/or the PD-L1 immune checkpoint inhibitor may be highly reactive. That is, as a result of performing a companion diagnosis to predict the reactivity of a PD-1 immune checkpoint inhibitor and/or a PD-L1 immune checkpoint inhibitor to cancer cells through the companion diagnostic biomarker composition according to the present invention, the HRG, AZGP1, complement C5, When the protein amount of any one or more biomarkers selected from the group consisting of IHKV4-1, MAN1A, A2M and HYOU1 is less than the cutoff value, the PD-1 immune checkpoint inhibitor and / or PD-L1 immune checkpoint inhibitor Reactivity means high. In this case, the PD-1 immune checkpoint inhibitor and/or the PD-L1 immune checkpoint inhibitor may be classified as a reactive patient group that is effective for chemotherapy.

Meanwhile, the biomarker composition for companion diagnosis according to the present invention is characterized in that it is extracted from any one selected from the group consisting of blood, serum, plasma and tissue. In addition, as an effect of the present invention, the present invention is capable of detecting the reactivity of any one or more immune checkpoint inhibitors among PD-1 immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors through blood proteomic analysis as well as co-diagnosis through existing tissues. It can be said that it can be predicted as one feature of the present invention.

That is, the biomarker composition for companion diagnosis according to the present invention is preferably extracted from blood, but is not limited thereto. More specifically, the biomarker composition for companion diagnosis according to the present invention performs a companion diagnosis through proteomic analysis on the blood of a cancer patient. has the effect of predicting the treatment response of

In addition, the biomarker composition for companion diagnosis according to the present invention may be extracted from a tissue as described above. In this case, the protein expression level of complement C7 can be measured through quantitative analysis. On the other hand, when the biomarker composition for companion diagnosis according to the present invention is extracted from the tissue, the complement C7 is compared to the non-responsive patient group that does not show an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor through the PD-1 immune checkpoint inhibitor. It is characterized in that the expression level of the protein is reduced in the responding patient group that is effective for anticancer treatment.

Conversely, when the biomarker composition for companion diagnosis according to the present invention is extracted from the tissue, the complement C7 is anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the reactive patient group showing the effect on anticancer treatment through the PD-1 immune checkpoint inhibitor It is characterized in that the expression level of the protein increases in the non-responding patient group that does not show an effect on

In addition, when the biomarker composition for companion diagnosis according to the present invention is extracted from the tissue, the complement C7 is compared to the non-responsive patient group that does not show an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor through the PD-L1 immune checkpoint inhibitor. It is characterized in that the expression level of the protein is reduced in the responding patient group that is effective for anticancer treatment.

Conversely, when the biomarker composition for companion diagnosis according to the present invention is extracted from a tissue, the complement C7 is anticancer treatment through PD-L1 immune checkpoint inhibitor compared to the reactive patient group showing the effect of anticancer treatment through PD-L1 immune checkpoint inhibitor It is characterized in that the expression level of the protein increases in the non-responding patient group that does not show an effect on

On the other hand, when the biomarker composition for companion diagnosis according to the present invention is extracted from a tissue, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is PD-1 immunity It is characterized in that the expression level of the protein is reduced in the responding patient group showing an effect on the anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the non-responding patient group showing no effect on the anticancer treatment through the checkpoint inhibitor.

Conversely, when the biomarker composition for companion diagnosis according to the present invention is extracted from tissue, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is PD-1 immunity It is characterized in that the expression level of the protein is increased in the non-responding patient group, which does not show an effect on chemotherapy through the PD-1 immune checkpoint inhibitor, compared to the responsive patient group that is effective for chemotherapy through the checkpoint inhibitor.

In addition, when the biomarker composition for companion diagnosis according to the present invention is extracted from a tissue, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is PD-L1 immune It is characterized in that the expression level of the protein is reduced in the responding patient group, which is effective for anticancer treatment through the PD-L1 immune checkpoint inhibitor, compared to the non-responding patient group, which does not show an effect on anticancer treatment through the checkpoint inhibitor.

Conversely, when the biomarker composition for companion diagnosis according to the present invention is extracted from a tissue, any one or more biomarkers selected from the group consisting of HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M and HYOU1 is PD-L1 immune It is characterized in that the expression level of the protein is increased in the non-responding patient group, which does not show an effect on chemotherapy through the PD-L1 immune checkpoint inhibitor, compared to the reactive patient group, which is effective for chemotherapy through the checkpoint inhibitor.

In addition, when the biomarker composition for companion diagnosis according to the present invention is extracted from a tissue, the biomarker composition for companion diagnosis may further include a Programmed Death-Ligand 1 (PD-L1) protein. Accordingly, treatment of one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor in cancer patients through the biomarker composition for companion diagnosis according to the present invention together with the measurement of the expression level of the PD-L1 protein reaction can be predicted.

On the other hand, in the biomarker composition for companion diagnosis according to the present invention, the cancer cells are cancer cells corresponding to carcinomas that specifically respond to at least one immune checkpoint inhibitor selected from a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor. do. That is, in the present invention, any target cancer that can be treated with cancer can be freely performed with any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor.

In addition, in the companion diagnostic biomarker composition according to the present invention, the cancer cells are lung cancer, liver cancer, gastric cancer, gastric and gastroesophageal junction adenocarcinoma, skin melanoma, head and neck cancer, bone cancer, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, rectal cancer, colon cancer, Colon cancer, breast cancer, uterine sarcoma, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, laryngeal cancer, small intestine cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, Any one selected from the group consisting of chronic or acute leukemia, solid tumors of childhood, differentiated lymphoma, bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma It is characterized in that it is a cancer cell corresponding to a carcinoma of

More specifically, in the biomarker composition for companion diagnosis according to the present invention, the cancer cell is preferably a cancer cell corresponding to lung cancer or liver cancer, but is not limited thereto.

On the other hand, the biomarker composition for companion diagnosis according to the present invention is characterized in that it is administered simultaneously or sequentially with one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor.

<Companion diagnostic kit>

Meanwhile, the present specification includes complement C7 (Complement Component C7), and any one or more of a PD-1 checkpoint inhibitor and a PD-L1 checkpoint inhibitor for cancer cells. Further disclosed is a companion diagnostic kit comprising a companion diagnostic biomarker composition characterized in predicting the reactivity of an immune checkpoint inhibitor. In the companion diagnosis kit according to the present invention, the above-mentioned <biomarker composition for companion diagnosis> applies mutatis mutandis to the biomarker composition for companion diagnosis.

The companion diagnostic kit according to the present invention may include the companion diagnostic biomarker composition according to the present invention.

The companion diagnostic kit according to the present invention is preferably any one kit selected from the group consisting of an ELISA (Enzyme Linked Immunosorbent Assay) kit, a protein chip kit, a Rapid kit, and a MRM (Multiple Reaction Monitoring) kit, but limited thereto it's not going to be In this case, any one kit selected from the group consisting of an ELISA kit, a protein chip kit, a Rapid kit, and a Multiple Reaction Monitoring (MRM) kit may be a kit performed by a known method.

In addition, the companion diagnostic kit according to the present invention may further include one or more other component compositions, solutions or devices suitable for the analysis method.

The companion diagnostic kit according to the present invention may be a diagnostic kit comprising essential elements necessary for performing ELISA. The ELISA kit may include an antibody that specifically binds to the protein or peptide fragment, an interacting protein, a ligand, nanoparticles, or an aptamer. The antibody is an antibody having high specificity and affinity for the protein of each complement and little cross-reactivity to other proteins, and may be a monoclonal antibody, a polyclonal antibody, or a recombinant antibody. In addition, the ELISA kit may include an antibody specific for an interacting protein, ligand, nanoparticles, aptamer, or control protein that specifically binds to the protein or peptide fragment. In addition, the ELISA kit includes a reagent capable of detecting bound antibody, for example, a labeled secondary antibody, chromophores, an enzyme (eg, an enzyme conjugated with an antibody) and a substrate thereof or the above. It may include other substances capable of binding to the antibody, and the like.

Hereinafter, with reference to the accompanying drawings and embodiments will be described in more detail with respect to what the present specification claims. However, the drawings and examples presented in this specification may be modified in various ways by those skilled in the art to have various forms, and the description of the present specification is not intended to limit the present invention to a specific disclosed form. It should be regarded as including all equivalents or substitutes included in the spirit and scope of the present invention. In addition, the accompanying drawings are presented to help those of ordinary skill in the art to more accurately understand the present invention, and may be exaggerated or reduced than in reality.

{Examples and Evaluation}

Example 1. Selection of 9 types of companion diagnostic biomarkers including complement C7

9 biomarker candidates included in the biomarker composition for companion diagnosis according to the present invention were selected through data mining, and validation stage was performed on the selected 9 biomarker candidates.

10 types of blood samples (hereinafter referred to as “ining set”) for which the reactivity of PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor to cancer cells of lung cancer patients were determined were prepared. Here, the 10 types of blood samples include 8 types of blood samples from a responder group showing an effect on chemotherapy through a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor and a PD-1 immune checkpoint inhibitor or PD-L1 There were two types of blood samples from a non-responder (Progressive Disease, PD) group that did not show an effect on chemotherapy with an immune checkpoint inhibitor. In addition, 8 blood samples from the responder group that showed effects on chemotherapy through PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor were found to be anticancer through PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor. Five blood samples from the reactive patient group (Partial Response, PR) showing high therapeutic efficacy and the low response patient group (Stable Disease, SD) with PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor of blood samples were classified into three types.

Table 1 below shows specific information on 10 blood samples for which the reactivity of the PD-1 immune checkpoint inhibitor or the PD-L1 immune checkpoint inhibitor has already been determined for cancer cells of lung cancer patients. However, in Table 1 below, PD means a non-responding patient group that does not show an effect on chemotherapy through a PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor, and PR means a PD-1 immune checkpoint inhibitor or PD-L1 immune It refers to a group of responsive patients that show a high effect on chemotherapy through a checkpoint inhibitor, and SD refers to a group of responsive patients showing a low effect on chemotherapy through a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor. In addition, in Table 1 below, M represents male and F represents female.

No. group
(Group) gender
(Gender) age
(Age) immune checkpoint inhibitors One PR M 52 Nivolumab (PD-1 Checkpoint Inhibitor) 2 F 69 Pembrolizumab (PD-1 Checkpoint Inhibitor) 3 M 62 Atezolizumab (PD-L1 Checkpoint Inhibitor) 4 M 58 Nivolumab (PD-1 Checkpoint Inhibitor) 5 M 65 Nivolumab (PD-1 Checkpoint Inhibitor) 6 SD M 60 Pembrolizumab (PD-1 Checkpoint Inhibitor) 7 F 58 Pembrolizumab (PD-1 Checkpoint Inhibitor) 8 F 71 Pembrolizumab (PD-1 Checkpoint Inhibitor) 9 PD M 69 Nivolumab (PD-1 Checkpoint Inhibitor) 10 M 70 Atezolizumab (PD-L1 Checkpoint Inhibitor)

To predict the reactivity of PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor to 10 blood samples for which the reactivity of PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor has already been determined for cancer cells in lung cancer patients , After extracting plasma from 10 types of blood samples, each of them undergoes a pretreatment process, and protein analysis is performed on the 10 types of plasma using TMT10 Isobaric Compounds, and the selected biomarker candidates 9 The amount of protein for each species was determined. However, in the protein analysis, the amounts of proteins for each of the nine biomarker candidates were measured through quantitative analysis.

At this time, the cutoff value of the protein amount measured for each of the nine biomarker candidates is set to 100 μg/mL, respectively, and when the protein amount of the nine biomarker candidates is equal to or greater than the cutoff value, the PD-1 immune checkpoint inhibitor or PD- If the amount of protein of 9 biomarker candidates is less than the cutoff value, the biomarker candidate 9 Depending on the amount of protein measured in the species, the reactive patient group (Partial Response, PR) and PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor, which show a high effect on chemotherapy with PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor It was classified into a reactive patient group (Stable Disease, SD) showing a low effect on anticancer treatment through an inhibitor (hereinafter referred to as 'Example 1').

Evaluation 1. Efficacy evaluation of a companion diagnostic biomarker composition containing complement C7

1 is a graph showing the results of a companion diagnosis predicting the reactivity of any one or more immune checkpoint inhibitors of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor against 9 types of companion diagnostic biomarkers including complement C7. it will be shown More specifically, Fig. 1a is biomarker C7, Fig. 1b is biomarker HRG, Fig. 1c is biomarker AZGP1, Fig. 1d is biomarker C5, Fig. 1e is biomarker IGKV4-1, Fig. 1f is biomarker MAN1A, Fig. 1g is a biomarker A2M, FIG. 1h is a biomarker SPP1, and FIG. 1i is a graph showing the results of a companion diagnosis predicting the reactivity of a PD-1 immune checkpoint inhibitor to cancer cells with respect to the biomarker HYOU1.

Referring to FIG. 1 , as a result of protein analysis for 10 blood samples, the measured amount of biomarker protein is constant even after several days have elapsed, and when the biomarker protein amount is equal to or greater than a cutoff value, and when the biomarker protein amount is greater than the cutoff value It can be confirmed that the companion diagnosis result of biomarker C7, which shows a certain tendency, is the best when it is less than. Accordingly, biomarker C7 was specified as the biomarker composition for companion diagnosis according to the present invention, and the remaining biomarkers HRG, AZGP1, C5, IHKV4-1, MAN1A, A2M and HYOU1 were used in combination with C7. As a biomarker composition according to the present invention, it can be estimated that it will show excellent companion diagnostic results.

Example 2. Companion diagnostic test using biomarker composition for companion diagnosis according to the present invention

According to the results of Example 1, 11 types of blood samples (hereinafter, referred to as “Set”) of lung cancer patients were prepared, plasma was extracted from each of the 11 types of blood samples, and then subjected to pretreatment, TMT10 isobaric compound (TMT10 Isobaric) Compounds) were used to perform protein analysis on 11 types of plasma to measure the amount of complement C7 protein, respectively. However, in the protein analysis, the amount of the complement C7 protein was measured through quantitative analysis, respectively.

Specific information on 11 blood samples from lung cancer patients is shown in Table 2 below. However, in Table 2 below, PD means a non-responding patient group that does not show an effect on chemotherapy through a PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor, and PR means a PD-1 immune checkpoint inhibitor or PD-L1 immune It refers to a group of responsive patients showing a high effect on chemotherapy through a checkpoint inhibitor, and SD refers to a group of responsive patients showing a low effect on chemotherapy through a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor. In addition, in Table 2 below, M represents male and F represents female.

No. group
(Group) gender
(Gender) age
(Age) immune checkpoint inhibitors 11 PR M 59 Pembrolizumab (PD-1 Checkpoint Inhibitor) 12 F 85 Pembrolizumab (PD-1 Checkpoint Inhibitor) 13 SD M 65 Pembrolizumab (PD-1 Checkpoint Inhibitor) 14 M 47 Nivolumab (PD-1 Checkpoint Inhibitor) 15 M 71 Nivolumab (PD-1 Checkpoint Inhibitor) 16 PD F 61 Pembrolizumab (PD-1 Checkpoint Inhibitor) 17 M 66 Pembrolizumab (PD-1 Checkpoint Inhibitor) 18 M 62 Nivolumab (PD-1 Checkpoint Inhibitor) 19 M 45 Nivolumab (PD-1 Checkpoint Inhibitor) 20 M 75 Atezolizumab (PD-L1 Checkpoint Inhibitor) 21 M 55 Pembrolizumab (PD-1 Checkpoint Inhibitor)

At this time, the cutoff value of the protein amount measured for complement C7 is set to 100 μg/mL, and when the protein amount of complement C7 is greater than or equal to the cutoff value, anticancer through PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor It is classified as a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect to treatment, and when the complement C7 protein amount is less than the cutoff value, the PD-1 immune checkpoint inhibitor according to the measured complement C7 protein amount Alternatively, the reactive patient group (Partial Response, PR), which shows a high effect on chemotherapy through PD-L1 immune checkpoint inhibitor, and the reactive patient group (Stable Disease, SD) (hereinafter referred to as 'Example 2').

Evaluation 2. Analysis and evaluation of companion diagnostic test results through the biomarker composition for companion diagnosis according to the present invention

2 is a graph showing the results of a companion diagnostic test using the biomarker composition for companion diagnostics according to an embodiment of the present invention. More specifically, FIG. 2A shows the results of a companion diagnostic test through biomarker C7 in the training set of Example 1, and FIG. 2B shows the results of a companion diagnostic test through complement C7 in the validation set of Example 2. , FIG. 2C shows the final result of combining the companion diagnostic test result through the biomarker C7 in the training set of Example 1 and the companion diagnostic test result through the complement C7 in the validation set of Example 2 above.

Referring to FIG. 2 , as a result of a companion diagnostic test using the biomarker composition for companion diagnosis according to the present invention for 11 blood samples of lung cancer patients of Example 2, PD based on the cutoff value according to the protein amount of complement C7 -1 Non-responders (Non-Responder; Progressive Disease, PD) that do not show an effect on chemotherapy through immune checkpoint inhibitors or PD-L1 immune checkpoint inhibitors, PD-1 immune checkpoint inhibitors or PD-L1 immune checkpoint inhibitors It can be accurately classified into a reactive patient group (Partial Response, PR), which shows a high effect on chemotherapy, and a reactive patient group (Stable Disease, SD), which shows a low effect on chemotherapy with a PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor. can check what is possible.

That is, since the biomarker composition for companion diagnosis according to the present invention contains complement C7, it can be said that it has an excellent effect in companion diagnosis for predicting the reactivity of a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor to cancer cells.

Example 3. Companion diagnostic test using the biomarker composition for companion diagnosis according to the present invention further comprising the biomarker IGKV4-1

10 blood samples (hereinafter, referred to as “ining set”) for which the reactivity of the PD-1 immune checkpoint inhibitor or the PD-L1 immune checkpoint inhibitor to cancer cells of a lung cancer patient of Example 1 were determined were prepared. Information on the 10 blood samples is the same as described above in Example 1. To predict the reactivity of PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor to 10 blood samples for which the reactivity of PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor has already been determined for cancer cells in lung cancer patients , After extracting plasma from 10 types of blood samples, each of them undergoes a pretreatment process, and protein analysis is performed on the 10 types of plasma using TMT10 Isobaric Compounds to perform biomarker complement C7 and IGKV4 The amount of protein of -1 (Immunoglobulin Kappa Variable 4-1) was measured, respectively. However, in the protein analysis, the amounts of proteins for the biomarkers complement C7 and IGKV4-1 were measured through quantitative analysis, respectively.

At this time, the relative ratio was obtained by dividing the measured complement C7 protein amount in each of 10 plasma types by the measured IGKV4-1 protein amount. According to the relative ratio, a non-responder (Non-Responder; Progressive Disease, PD), PD-1 immune checkpoint inhibitor or PD- The reactive patient group (Partial Response, PR), which shows a high effect on chemotherapy through L1 immune checkpoint inhibitor, and the reactive patient group (Stable Disease, SD) (hereinafter referred to as 'Example 3').

Evaluation 3. Companion diagnostic test analysis and evaluation using the biomarker composition for companion diagnosis according to the present invention further comprising the biomarker IGKV4-1

3 is a graph showing a companion diagnostic test result when the biomarker IGKV4-1 is further included in the companion diagnostic biomarker composition according to an embodiment of the present invention.

Referring to FIG. 3 , as a result of a companion diagnostic test using the biomarker composition for companion diagnosis according to the present invention for 10 blood samples of lung cancer patients of Example 3, the complement C7 protein amount was determined as the IGKV4-1 protein amount. Non-responder (Progressive Disease, PD), PD-1 immunity that does not show an effect on chemotherapy through PD-1 immune checkpoint inhibitor or PD-L1 immune checkpoint inhibitor by a constant reference value according to the relative ratio Response patient group (Partial Response, PR) showing high efficacy to anticancer treatment using checkpoint inhibitors or PD-L1 immune checkpoint inhibitors and responses showing low efficacy to chemotherapy using PD-1 immune checkpoint inhibitors or PD-L1 immune checkpoint inhibitors It can be confirmed that it can be accurately classified into a patient group (Stable Disease, SD). That is, the results according to Example 3 are consistent with the information about 10 blood samples listed in Table 1 above of Example 1.

That is, the biomarker composition for companion diagnosis according to the present invention contains complement C7, and thus has an excellent effect in companion diagnosis for predicting the reactivity of a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor to cancer cells. It was found that even when the biomarker IGKV4-1 is further included in the biomarker composition for companion diagnosis according to the present invention, there is a better effect in companion diagnosis for predicting the reactivity of a PD-1 immune checkpoint inhibitor or a PD-L1 immune checkpoint inhibitor to cancer cells. can be checked

Such results were obtained not only when the biomarker IGKV4-1 was further included in the complement C7, but also in the case of the complement C7 including Histidine-Rich Glycoprotein (HRG), Zinc-Alpha-2-Glycoprotein (AZGP1), and Complement Component C5 (C5). ), MAN1A (Mannosyl-Oligosaccharide 1,2-Alpha-Mannosidase 1A), A2M (Alpha-2-Macroglobulin), SPP1 (Osteopontin) and HYOU1 (Hypoxia Up-Regulated Protein 1) Even when the marker is further included, it can be estimated that the excellent companion diagnosis result as in Example 3 will be obtained. This is because HRG, AZGP1, complement C5, IHKV4-1, MAN1A, A2M, and HYOU1 are biomarkers that regulate the human antigen-antibody immune response and are involved in immune response and lysis according to gene ontology classification. It can be inferred from the common point of protein.

Experimental Example 1. When the cut-off value for the measured amount of complement C7 protein in the biomarker composition for companion diagnosis according to the present invention was 100 μg/mL (compared to the PD-L1 pharmDx test)

After preparing 27 types of blood samples from lung cancer patients, extracting plasma from each of the 27 types of blood samples, pre-processing, and analyzing the 27 types of plasma using TMT10 Isobaric Compounds (Proteome Analysis) to measure the amount of protein for complement C7, respectively. However, in the protein analysis, the amount of the complement C7 protein was measured through quantitative analysis, respectively.

At this time, the cutoff value of the protein amount measured for complement C7 is set to 100 μg/mL, and when the protein amount of complement C7 is greater than or equal to the cutoff value, any one of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor Classified as a non-responder (Progressive Disease, PD) group that does not show an effect on anticancer treatment through the above immune checkpoint inhibitors, and when the amount of complement C7 protein is less than the cutoff value, the measured amount of complement C7 protein Accordingly, the PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, which show a high effect on chemotherapy through one or more immune checkpoint inhibitors (Partial Response, PR), and PD-1 immune checkpoint inhibitor and PD-L1 immunity It was classified into a reactive patient group (Stable Disease, SD) showing a low effect on anticancer treatment through any one or more immune checkpoint inhibitors (hereinafter, referred to as 'Experimental Example 1').

Experimental Example 2. When the cut-off value for the measured amount of complement C7 protein in the companion diagnostic biomarker composition according to the present invention was 105 μg/mL (compared to the PD-L1 pharmDx test)

Except that the cutoff value of the protein amount measured for complement C7 was set to 105 μg/mL, an experiment was performed in the same manner as in Experimental Example 1, and when the protein amount of complement C7 was greater than or equal to the cutoff value, PD-1 Immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors are classified into a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors, and the protein amount of complement C7 is If it is less than the cutoff value, a reactive patient group (Partial Response, PR) that shows a high effect on anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor according to the measured amount of complement C7 protein (Partial Response, PR) ) and PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, classified into a reactive patient group (Stable Disease, SD) that shows a low effect on anticancer treatment through any one or more immune checkpoint inhibitors (hereinafter referred to as 'Experimental Example 2') did).

Experimental Example 3. When the cut-off value for the measured amount of complement C7 protein in the biomarker composition for companion diagnosis according to the present invention was 110 μg/mL (compared to the PD-L1 pharmDx test)

Except for setting the cutoff value of the protein amount measured for complement C7 to 110 μg/mL, an experiment was performed in the same manner as in Experimental Example 1, and when the protein amount of complement C7 was greater than or equal to the cutoff value, PD-1 Immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors are classified into a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors, and the protein amount of complement C7 is If it is less than the cutoff value, a reactive patient group (Partial Response, PR) that shows a high effect on anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor according to the measured amount of complement C7 protein (Partial Response, PR) ) and PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, classified into a reactive patient group (Stable Disease, SD) that shows a low effect on anticancer treatment through any one or more immune checkpoint inhibitors (hereinafter referred to as 'Experimental Example 3') did).

Comparative Experimental Example 1. Companion diagnostic test through “PD-L1 pharmDx test”

The PD-L1 pharmDx test (hereinafter, referred to as 'pharmDx') is a companion diagnostic test used to determine whether pembrolizumab effectively exhibits anticancer effects as an immunotherapy for specific patients.

27 kinds of tissue samples from lung cancer patients of Experimental Example 1 were prepared, and cancer cells in biopsied tissue samples using DAKO's PD-L1 pharmDx product were subjected to immunohistochemical staining using PD-L1 (Programmed Cell Death) (Programmed Cell Death). Protein 1 Ligand) protein was qualitatively tested to observe the expression rate of PD-L1 protein in the cancer cells. After the PD-L1 pharmDx test, the stained slides are observed through an optical microscope to calculate the Tumor Proportion Score (TPS), which is the percentage of effective tumor cells showing partial or total cell membrane staining of tumor cells, and TPS ≥ 50% In the case of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, it is classified as a reactive patient group (Partial Response, PR) that shows a high effect on chemotherapy through any one or more immune checkpoint inhibitors, and when the TPS is 1 to 49% It is classified as a reactive patient group (Stable Disease, SD) that shows a low effect on chemotherapy through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, and if TPS < 1%, PD-1 Immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors are classified into a non-responder group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors (hereinafter, 'Comparative Experimental Example 1') is called).

Evaluation 4. Results analysis and evaluation according to Experimental Examples 1 to 3 and Comparative Experimental Example 1

The following is a comparison and evaluation of the companion diagnostic test results according to Experimental Examples 1 to 3 and Comparative Experimental Example 1 described above.

Table 3 below shows the results of the companion diagnostic test according to Experimental Examples 1 to 3 and Comparative Experimental Example 1. In Table 3 below, “result” refers to the number of patients classified into the corresponding patient group (PR, PR+SD, PD) as a result of the experiment, and “actual” refers to the actual patient group (PR, PR+) regardless of whether or not the experiment was conducted. SD, PD) means the number of patients classified. That is, “result/actual” refers to the ratio of the number of patients classified into the corresponding patient group as a result of the experiment to the number of patients actually classified into the corresponding patient group.

Comparative Experimental Example 1 Experimental Example 1 Experimental Example 2 Experimental Example 3 percentage(%) result/actual percentage(%) result/actual percentage(%) result/actual percentage(%) result/actual PR 33 2/6 100 6/6 100 6/6 100 6/6 PR + SD 38 5/13 85 11/13 92 12/13 100 13/13 PD 100 8/8 75 6/8 75 6/8 63 5/8

Referring to Table 3, in Comparative Experimental Example 1 corresponding to the PD-L1 pharmDx test, the PD-1 immune checkpoint inhibitor and the PD-L1 immune checkpoint inhibitor show a high effect on anticancer treatment through any one or more immune checkpoint inhibitors. Accuracy of discriminating between the reactive patient group (Partial Response, PR) and the reactive patient group (Stable Disease, SD), which show low efficacy in chemotherapy with one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor It can be seen that is relatively lower than in Experimental Examples 1 to 3.

On the other hand, in the case of Experimental Examples 1 to 3, as a result of the companion diagnostic test through the companion diagnostic biomarker composition according to the present invention, the anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor. Although the accuracy of discriminating the non-responder group (Non-Responder; Progressive Disease, PD) that does not show an effect tends to be relatively low compared to Comparative Experimental Example 1, among PD-1 immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors, Partial Response (PR), which shows a high effect on chemotherapy with one or more immune checkpoint inhibitors It can be seen that the accuracy of discriminating the reactive patient group (Stable Disease, SD) showing the effect is significantly higher than that of Comparative Experimental Example 1.

Therefore, in the case of Experimental Examples 1 to 3, it can be confirmed that the companion diagnostic test result is a companion diagnostic test with better accuracy than Comparative Experimental Example 1 corresponding to the PD-L1 pharmDx test through the companion diagnostic biomarker composition according to the present invention. .

Experimental Example 4. When the cut-off value for the measured amount of complement C7 protein in the biomarker composition for companion diagnosis according to the present invention was 100 μg/mL (compared to SP263)

36 types of blood samples from lung cancer patients are prepared, plasma is extracted from each of the 36 types of blood samples, and then subjected to a pretreatment process. Protein analysis of the 36 types of plasma using TMT10 Isobaric Compounds (Proteome Analysis) to measure the amount of protein for complement C7, respectively. However, in the protein analysis, the amount of the complement C7 protein was measured through quantitative analysis, respectively.

At this time, the cutoff value of the protein amount measured for complement C7 is set to 100 μg/mL, and when the protein amount of complement C7 is greater than or equal to the cutoff value, any one of a PD-1 immune checkpoint inhibitor and a PD-L1 immune checkpoint inhibitor Classified into a non-responder; Progressive Disease, PD) that does not show an effect on anticancer treatment through the above immune checkpoint inhibitors, and when the amount of complement C7 protein is less than the cutoff value, the measured amount of complement C7 protein Accordingly, the PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, which show a high effect on chemotherapy through one or more immune checkpoint inhibitors (Partial Response, PR), and PD-1 immune checkpoint inhibitor and PD-L1 immunity The checkpoint inhibitors were classified as a reactive patient group (Stable Disease, SD) showing a low effect on anticancer treatment through one or more immune checkpoint inhibitors (hereinafter referred to as 'Experimental Example 4').

Experimental Example 5. When the cut-off value for the measured amount of complement C7 protein in the companion diagnostic biomarker composition according to the present invention was 105 μg/mL (compared to SP263)

When the protein amount of complement C7 is greater than or equal to the cutoff value in the same manner as in Experimental Example 4, except that the cutoff value of the protein amount measured for complement C7 is set to 105 μg/mL, PD-1 Immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors are classified into a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors, and the protein amount of complement C7 is When it is less than the cutoff value, a reactive patient group (Partial Response, PR) that shows a high effect on anticancer treatment through one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor according to the measured amount of complement C7 protein (Partial Response, PR) ) and PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, classified into a reactive patient group (Stable Disease, SD) that shows a low effect on anticancer treatment through any one or more immune checkpoint inhibitors (hereinafter referred to as 'Experimental Example 5') did).

Experimental Example 6. When the cut-off value for the measured amount of complement C7 protein in the biomarker composition for companion diagnosis according to the present invention was 110 μg/mL (compared to SP263)

When the protein amount of complement C7 is greater than or equal to the cutoff value in the same manner as in Experimental Example 4, except that the cutoff value of the protein amount measured for complement C7 is set to 110 μg/mL, PD-1 Immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors are classified into a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors, and the protein amount of complement C7 is When it is less than the cutoff value, a reactive patient group (Partial Response, PR) that shows a high effect on anticancer treatment through one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor according to the measured amount of complement C7 protein (Partial Response, PR) ) and PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, classified into a reactive patient group (Stable Disease, SD) showing a low effect on anticancer treatment through any one or more immune checkpoint inhibitors (hereinafter referred to as 'Experimental Example 6') did).

Comparative Experimental Example 2. Companion diagnostic test through “VENTANA PD-L1 (SP263) Assay”

The VENTANA PD-L1 (SP263) Assay (hereinafter referred to as 'SP263') is a companion diagnostic test used to determine whether nivolumab effectively exhibits anticancer effects as an immunotherapy for specific patients.

By preparing 36 tissue samples from lung cancer patients of Experimental Example 1, and using the VENTANA PD-L1 (SP263) Assay product of VENTANA, the cancer cells in the biopsied tissue sample were PD- L1 (Programmed Cell Death Protein 1 Ligand) protein was qualitatively tested to observe the expression rate of PD-L1 protein in the cancer cells. After the PD-L1 pharmDx test, the stained slides are observed through an optical microscope to calculate the Tumor Proportion Score (TPS), which is the percentage of effective tumor cells showing partial or total cell membrane staining of tumor cells, and TPS ≥ 10% In the case of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, it is classified as a reactive patient group (Partial Response, PR) that shows a high effect on cancer treatment through any one or more immune checkpoint inhibitors, and when the TPS is 1 to 9% It is classified as a reactive patient group (Stable Disease, SD) that shows a low effect on chemotherapy through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, and if TPS < 1%, PD-1 Classified as a non-responder (Progressive Disease, PD) group (hereinafter, 'Comparative Experimental Example 2') that does not show an effect on anticancer treatment through any one or more immune checkpoint inhibitors of immune checkpoint inhibitors and PD-L1 immune checkpoint inhibitors is called).

Evaluation 5. Analysis and evaluation of results according to Experimental Examples 4 to 6 and Comparative Experimental Example 2

The following is a comparison and evaluation of the companion diagnostic test results according to Experimental Examples 4 to 6 and Comparative Experimental Example 2 described above.

Table 4 below shows the results of the companion diagnostic test according to Experimental Examples 4 to 6 and Comparative Experimental Example 2. In Table 4 below, “result” means the number of patients classified into the corresponding patient group (PR, PR+SD, PD) as a result of the experiment, and “actual” means the actual patient group (PR, PR+) regardless of whether or not the experiment was conducted. SD, PD) means the number of patients classified. That is, “result/actual” refers to the ratio of the number of patients classified into the corresponding patient group as a result of the experiment to the number of patients actually classified into the corresponding patient group.

Comparative Experimental Example 2 Experimental Example 4 Experimental Example 5 Experimental Example 6 percentage(%) result/actual percentage(%) result/actual percentage(%) result/actual percentage(%) result/actual PR 67 6/9 100 9/9 100 9/9 100 9/9 PR + SD 71 12/17 76 13/17 88 15/17 100 17/17 PD 80 8/10 90 9/10 90 9/10 70 7/10

Referring to Table 4 above, in the case of Comparative Experimental Example 2 corresponding to the VENTANA PD-L1 (SP263) Assay, high anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor The reactive patient group (Partial Response, PR) and PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor (PD-L1 immune checkpoint inhibitor) showed a low effect on chemotherapy with one or more immune checkpoint inhibitors (Stable Disease, SD). It can be seen that the determination accuracy is relatively lower than those of Experimental Examples 4 to 6. In addition, the accuracy of discriminating the non-responder (Progressive Disease, PD) group that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor is also It can be confirmed that it is relatively low compared to Experimental Examples 4 to 6.

Conversely, in the case of Experimental Examples 4 to 6, the companion diagnostic test showed a high level of anticancer treatment through one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor through the companion diagnostic biomarker composition according to the present invention. Response patient group showing efficacy (Partial Response, PR), PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor, responding patient group showing low effect on chemotherapy with one or more immune checkpoint inhibitors (Stable Disease, SD) and The accuracy of discriminating the non-responder (Progressive Disease, PD) of a non-responder patient group (Non-Responder; Progressive Disease, PD) that does not show an effect on chemotherapy through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor is a comparative experimental example It can be seen that it is relatively higher than that of 2.

Therefore, in the case of Experimental Examples 4 to 6, the companion diagnostic test result through the companion diagnostic biomarker composition according to the present invention is a companion diagnostic test with better accuracy than Comparative Experimental Example 2 corresponding to the VENTANA PD-L1 (SP263) Assay. can be checked

The biomarker composition for companion diagnosis of the present invention and the companion diagnostic kit comprising the same according to the present invention as described above are not only a companion diagnosis through the tissue of a cancer patient, but also a PD-1 immune checkpoint inhibitor and PD-L1 through proteomic analysis in the blood of a cancer patient. There is an effect of predicting the therapeutic response of any one or more immune checkpoint inhibitors among the immune checkpoint inhibitors.

In addition, the biomarker composition for companion diagnosis and the companion diagnosis kit comprising the same according to the present invention include a reactive patient group and It has the advantage of high accuracy in screening non-responding patients who do not show the effect.

Furthermore, the biomarker composition for companion diagnosis and the companion diagnosis kit comprising the same according to the present invention more accurately determine a patient group suitable for anticancer treatment through any one or more immune checkpoint inhibitors of PD-1 immune checkpoint inhibitor and PD-L1 immune checkpoint inhibitor There is an effect that can be done.

The above description is merely illustrative of the technical spirit of the present invention, and various modifications and variations will be possible without departing from the essential characteristics of the present invention by those skilled in the art to which the present invention pertains.

Accordingly, the embodiments disclosed in the present invention are not intended to limit the technical spirit of the present invention, but to explain, and the scope of the technical spirit of the present invention is not limited by these embodiments. The protection scope of the present invention should be construed by the following claims, and all technical ideas within the scope equivalent thereto should be construed as being included in the scope of the present invention.

Claims (22)

Complement C7 (Complement Component C7), and the reactivity of any one or more immune checkpoint inhibitors of PD-1 (Programmed Cell Death Protein 1) immune checkpoint inhibitors and PD-L1 (Programmed Death-Ligand 1) immune checkpoint inhibitors against cancer cells As a biomarker composition for companion diagnosis, characterized in that predicting
The complement C7 is extracted from blood,
The composition is a biomarker composition for companion diagnosis, characterized in that it predicts the reactivity of an immune checkpoint inhibitor to cancer cells by quantitatively analyzing the amount of complement C7 protein extracted from the blood.
According to claim 1,
The companion diagnostic biomarker composition is HRG (Histidine-Rich Glycoprotein), AZGP1 (Zinc-Alpha-2-Glycoprotein), complement C5 (Complement Component C5), IHKV4-1 (Immunoglobulin Kappa Variable 4-1), MAN1A (Mannosyl- Oligosaccharide 1,2-Alpha-Mannosidase 1A), A2M (Alpha-2-Macroglobulin), SPP1 (Osteopontin) and HYOU1 (Hypoxia Up-Regulated Protein 1) further comprising any one or more biomarkers selected from the group consisting of, A biomarker composition for companion diagnosis.
delete According to claim 1,
The amount of complement C7 protein was determined by protein mass spectrometry, protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry analysis, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry (Liquid Chromatography-) Mass Spectrometry, LC-MS), LC-MS/MS (Liquid Chromatography-Mass Spectrometry/ Mass Spectrometry), Western blot, and ELISA (Enzyme Linked Immunosorbent Assay), characterized in that measured by any one method selected from the group consisting of , Biomarker composition for companion diagnosis.
According to claim 1,
The complement C7 decreases the amount of protein in the reactive patient group showing an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor A biomarker composition for companion diagnosis, characterized in that.
According to claim 1,
The complement C7 increases the amount of protein in the non-responding patient group that does not show an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor compared to the reactive patient group that shows an effect on anticancer treatment through the PD-1 immune checkpoint inhibitor A biomarker composition for companion diagnosis, characterized in that.
According to claim 1,
The complement C7 protein is decreased in the reactive patient group showing an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor compared to the non-responding patient group that does not show an effect on anticancer treatment through the PD-L1 immune checkpoint inhibitor A biomarker composition for companion diagnosis, characterized in that.
According to claim 1,
The complement C7 increases the amount of protein in the non-responsive patient group that does not show an effect on chemotherapy through the PD-L1 immune checkpoint inhibitor compared to the responsive patient group that is effective for anticancer treatment through the PD-L1 immune checkpoint inhibitor A biomarker composition for companion diagnosis, characterized in that.
According to claim 1,
A biomarker composition for companion diagnosis, characterized in that the cutoff value for the protein amount of complement C7 is within the range of 100 to 110 μg/mL.
10. The method of claim 9,
A biomarker composition for companion diagnosis, characterized in that the reactivity of the PD-1 immune checkpoint inhibitor to the cancer cells is low when the amount of the complement C7 protein is equal to or greater than the cutoff value.
10. The method of claim 9,
When the amount of the complement C7 protein is less than the cutoff value, the biomarker composition for companion diagnosis, characterized in that the reactivity of the PD-1 immune checkpoint inhibitor to the cancer cells is high.
10. The method of claim 9,
A biomarker composition for companion diagnosis, characterized in that when the amount of the complement C7 protein is equal to or greater than the cutoff value, the reactivity of the PD-L1 immune checkpoint inhibitor to the cancer cells is low.
10. The method of claim 9,
A biomarker composition for companion diagnosis, characterized in that the reactivity of the PD-L1 immune checkpoint inhibitor to the cancer cells is high when the amount of the complement C7 protein is less than the cutoff value.
delete delete delete According to claim 1,
The biomarker composition for companion diagnosis further comprises a PD-L1 (Programmed Death-Ligand 1) protein.
According to claim 1,
The cancer cell is a cancer cell corresponding to a carcinoma that specifically responds to at least one immune checkpoint inhibitor of the PD-1 immune checkpoint inhibitor and the PD-L1 immune checkpoint inhibitor, a companion diagnostic biomarker composition.
According to claim 1,
The cancer cells are lung cancer, liver cancer, stomach cancer, stomach and gastroesophageal junction adenocarcinoma, skin melanoma, head and neck cancer, bone cancer, pancreatic cancer, skin cancer, uterine cancer, ovarian cancer, rectal cancer, colorectal cancer, colon cancer, breast cancer, uterine sarcoma, fallopian tube carcinoma, endometrial cancer Carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, esophageal cancer, larynx cancer, small intestine cancer, thyroid cancer, parathyroid cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, solid tumors in childhood, differentiated lymphoma , bladder cancer, kidney cancer, renal cell carcinoma, renal pelvic carcinoma, first central nervous system lymphoma, spinal cord tumor, brainstem glioma and pituitary adenoma, characterized in that the cancer cell corresponding to any one carcinoma selected from the group consisting of, A biomarker composition for companion diagnosis.
20. The method of claim 19,
Wherein the cancer cells are cancer cells corresponding to lung cancer or liver cancer, a biomarker composition for companion diagnosis.
According to claim 1,
The biomarker composition for companion diagnosis is characterized in that it is administered simultaneously or sequentially with at least one immune checkpoint inhibitor of the PD-1 immune checkpoint inhibitor and the PD-L1 immune checkpoint inhibitor.
A kit for companion diagnosis, comprising the biomarker composition for companion diagnosis according to any one of claims 1, 2, 4 to 13, and claims 17 to 21.

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